BIPHENYL-SUBSTITUTED EPITHELIAL SODIUM CHANNEL BLOCKING COMPOUNDS

Abstract

The present invention relates to ENaC inhibitors (e.g., compounds of Formula (I), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof). Also disclosed are compositions, methods of preparation, combination therapies, kits, uses, and methods. Exemplary uses include promoting hydration of mucosal surfaces and treating diseases and disorders including chronic obstructive pulmonary disease (COPD), asthma, bronchiectasis, acute and chronic bronchitis, cystic fibrosis, primary ciliary dyskinesia, idiopathic pulmonary fibrosis, and pneumonia.

##STR00001##

Claims

1. A compound of Formula (I): ##STR00547## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, wherein: R.sup.W1 is hydrogen, halogen, optionally substituted alkyl, or N(R.sup.N).sub.2 R.sup.W2 is hydrogen, halogen, optionally substituted alkyl, or N(R.sup.N).sub.2 optionally where R.sup.W1 and R.sup.W2 are joined together to form optionally substituted heterocyclyl or optionally substituted heteroaryl; each instance of R.sup.N is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a nitrogen protecting group, or optionally two instances of R.sup.N bonded to the same nitrogen atom are joined together to form optionally substituted heterocyclyl or optionally substituted heteroaryl; each instance of L.sup.1, L.sup.2, L.sup.2, and L.sup.4 is independently a bond, optionally substituted C.sub.1-10 alkylene, optionally substituted C.sub.2-10 alkenylene, optionally substituted C.sub.2-10 alkynylene, optionally substituted C.sub.1-10 heteroalkylene, optionally substituted C.sub.2-10heteroalkenylene, or optionally substituted C.sub.2-10 heteroalkynylene; each instance of R.sup.4 is independently halogen, CN, NO.sub.2, N.sub.3, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, OR.sup.O, N(R.sup.N).sub.2, or SR.sup.S; each instance of R.sup.O is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or an oxygen protecting group; each instance of R.sup.S is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a sulfur protecting group; each p is independently selected from 0, 1, 2, 3, and 4; each instance of Y.sup.1 and Y.sup.2 is independently a bond, CH.sub.2, O, S, NR.sup.8, C(O), S(O), S(O).sub.2, OC(O), OS(O).sub.2, C(O)O, S(O).sub.2O, NR.sup.8C(O), NR'S(O).sub.2, C(O)NR.sup.8, S(O).sub.2NR.sup.B, or ##STR00548## each instance of R.sup.8 is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a nitrogen protecting group, optionally wherein R.sup.8 is substituted with 0, 1, or 2 NRR.sup.1; Ring A is optionally substituted arylene, optionally substituted heteroarylene, optionally substituted heterocyclylene, or optionally substituted heteroarylene; Z is hydrogen, NRR.sup.1, N.sup.+(O.sup.)RR.sup.1, OR.sup.B, C(R.sup.1).sub.3, NR.sup.A(CO)R.sup.C, NR.sup.A(CO)OR.sup.B, NR.sup.A(CO)N(R.sup.A).sub.2, NR.sup.A(CNR.sup.A)N(R.sup.A).sub.2, (CO)OR.sup.B, (CO)N(R.sup.A).sub.2, or B; each instance of R.sup.A is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a nitrogen protecting group, optionally wherein two R.sup.A bonded to the name nitrogen atom are joined together to form optionally substituted heteroaryl or optionally substituted heterocyclyl; each instance of R.sup.B is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or an oxygen protecting group; each instance of R.sup.C is independently hydrogen or optionally substituted alkyl; each instance of R and R.sup.1 is independently hydrogen, optionally substituted alkyl, optionally substituted heteroalkyl, a polyhydroxylated alkyl group, a polyhydroxylated heteroalkyl group, optionally substituted acyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted -alkyl-E, optionally substituted -heteroalkyl-E, or a nitrogen protecting group, optionally wherein R and R.sup.1 bonded to the same nitrogen atom are joined together with the intervening atoms to form optionally substituted heterocyclyl or optionally substituted heteroaryl; each instance of E is independently optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl, optionally wherein E is a cyclic sugar; and B is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl, optionally wherein B is substituted with 0, 1, or 2 R.sup.1.

2. The compound of claim 1, wherein the compound is of Formula (II): ##STR00549## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

3. The compound of claim 1 or 2, wherein the compound is of Formula (III): ##STR00550## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, wherein: c is selected from 0, 1, 2, 3, 4, 5, and 6; X is selected from a bond, CH.sub.2, O, N(R.sup.N) and S; each n is independently selected from 0, 1, 2, 3, 4, 5, and 6; each instance of R.sup.2 and R.sup.3 is independently selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, N(R.sup.9).sub.2, OR.sup.9, C(O)OR.sup.9, C(O)N(R.sup.9).sub.2, NR.sup.AC(O)R.sup.9, NR.sup.AC(O)OR.sup.9, NR.sup.AC(O)N(R.sup.9).sub.2, OC(O)R.sup.9, OC(O)OR.sup.9, OC(O)N(R.sup.9).sub.2, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heterocyclyl, and optionally substituted heteroaryl, optionally wherein R.sup.2 and R.sup.3 is substituted with 0, 1, or 2 N(R.sup.A).sub.2, C(O)OR.sup.B, and NR.sup.AC(O)R.sup.10; each instance of R.sup.9 is independently selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, an amino acid, a peptide comprising 2, 3, 4, 5, or 6 amino acids, or a nitrogen or oxygen protecting group, optionally wherein two R.sup.9 bonded to the same nitrogen atom are joined together to form optionally substituted heteroaryl or optionally substituted heterocyclyl; and optionally wherein R.sup.9 is substituted with 0, 1, or 2 groups selected from N(R.sup.A).sub.2, C(O)OR.sup.10, and NR.sup.AC(O)R.sup.10; each instance of R.sup.10 is independently selected from hydrogen or optionally substituted alkyl substituted with 0, 1, or 2 groups selected from N(R.sup.A).sub.2, C(O)OR.sup.B, and NR.sup.AC(O)R.sup.C; u is selected from 0 and 1; and t is selected from 0 and 1.

4. The compound of claim 3, wherein the compound is of Formula (IV): ##STR00551## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

5. The compound of claim 3, wherein the compound is of Formula (V): ##STR00552## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

6. The compound of claim 3, wherein the compound is of one of the following formulae: ##STR00553## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, wherein D is O or NR.sup.8.

7. The compound of claim 6, wherein the compound is of one of the following formulae: ##STR00554## or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, wherein D is O or NR.sup.8.

8. The compound of any one of claims 3-7, wherein the compound is of one of the following formulae: ##STR00555## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, wherein D is O or NR.sup.8.

9. The compound of any one of claims 3-8, wherein the compound is of one of the following formulae: ##STR00556## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, wherein D is O or NR.sup.8.

10. The compound of claim 3, wherein the compound is of the formula: ##STR00557## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, wherein g is 0, 1, 2, 3, 4, or 5.

11. The compound of claim 10, wherein the compound is of the formula: ##STR00558## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

12. The compound of claim 11, wherein the compound is of the formula: ##STR00559## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative

13. The compound of claim 3, wherein the compound is of the formula: ##STR00560## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

14. The compound of claim 3, wherein the compound is of the formula: ##STR00561## or pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

15. The compound of claim 14, wherein the compound is of the formula: ##STR00562## or pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

16. The compound of claim 15, wherein the compound is of the formula: ##STR00563## or pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

17. The compound of claim 16, wherein the compound is of the formula: ##STR00564## or pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

18. The compound of claim 3, wherein the compound is of the formula: ##STR00565## or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

19. The compound of claim 3, wherein the compound is of the formula: ##STR00566## or pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof.

20. The compound of any one of the preceding claims, wherein Z is B.

21. The compound of any one of the preceding claims, wherein B is optionally substituted phenyl, optionally substituted pyridinyl, optionally substituted piperidinyl, or optionally substituted indolyl.

22. The compound of any one of the preceding claims, wherein B is phenyl, pyridinyl, piperidinyl, or indolyl with each phenyl, pyridinyl, piperidinyl, or indolyl ring substituted by 0, 1, 2, or 3 substituents independently selected from halogen, OR.sup.6, C.sub.1-6 alkyl substituted with 0 or 1 NRR.sup.1, and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms; wherein R.sup.6 is hydrogen, C.sub.1-6 alkyl substituted with 0 or 1 NRR.sup.1, or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms.

23. The compound of any one of the preceding claims, wherein B is substituted with 1 or 2 groups of the formula: ##STR00567##

24. The compound of any one of the preceding claims, wherein B is phenyl.

25. The compound of any one of the preceding claims, wherein Z is hydrogen, C(R.sup.1).sub.3, C(O)OR.sup.B, or NR.sup.A(CNR.sup.A)N(R.sup.A).sub.2.

26. The compound of any one of the preceding claims, wherein Z is NRR.sup.1.

27. The compound of any one of the preceding claims, wherein NRR.sup.1 is of the formula: ##STR00568##

28. The compound of any one of the preceding claims, wherein NRR.sup.1 is of the formula: ##STR00569##

29. The compound of any one of the preceding claims, wherein Ring A is piperdinylene or pyrrolidinylene.

30. The compound of any one of the preceding claims, wherein Ring A is selected from ##STR00570## wherein each R.sup.Y is independently hydrogen, halogen, O, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.O, N(R.sup.N).sub.2, SR.sup.S, optionally substituted acyl, C(O)OR.sup.O, or C(O)N(R.sup.N).sub.2; and y is selected from 0, 1, and 2.

31. The compound of any one of the preceding claims wherein y is 1 and R.sup.Y is O, C(O)OH or C(O)OCH.sub.3.

32. The compound of any one of the preceding claims, wherein R.sup.2 is a C.sub.1-6 alkyl substituted with N(R.sup.9).sub.2.

33. The compound of any one of the preceding claims, wherein R.sup.3 is a C.sub.1-6 alkyl substituted with N(R.sup.9).sub.2.

34. The compound of any one of the preceding claims, wherein one instance of R.sup.2 and R.sup.3 is independently of one of the following formulae: ##STR00571## or, or salt thereof.

35. The compound of any one of the preceding claims, wherein each instance of R.sup.2 and R.sup.3 is hydrogen.

36. The compound of any one of the preceding claims, wherein R.sup.8 is hydrogen or optionally substituted C.sub.1-6 alkyl.

37. The compound of any one of the preceding claims, wherein R.sup.8 is a C.sub.1-6 alkyl substituted with NRR.sup.1

38. The compound of any one of the preceding claims, wherein at least one instance of R.sup.9 is hydrogen or a C.sub.1-6 alkyl substituted with 0, 1, or 2 groups selected from N(R.sup.A).sub.2, C(O)OR.sup.10, and NR.sup.AC(O)R.sup.10.

39. The compound of any one of the preceding claims, wherein R.sup.9 is a C.sub.1-6 alkyl substituted with N(R.sup.A).sub.2

40. The compound of any one of the preceding claims, wherein R.sup.10 is independently hydrogen or C.sub.1-6alkyl substituted with N(R.sup.A).sub.2, C(O)OR.sup.B, and NR.sup.AC(O)R.sup.C.

41. The compound of any one of the preceding claims, wherein at least one instance of R.sup.9 is independently hydrogen, C.sub.1-6 alkyl, or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive.

42. The compound of any one of the preceding claims, wherein each R.sup.A is independently hydrogen, C.sub.1-6 alkyl, or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive.

43. The compound of any one of the preceding claims, wherein R.sup.A is hydrogen or C.sub.1-6 alkyl.

44. The compound of any one of the preceding claims, wherein each R.sup.B is independently hydrogen, C.sub.1-6 alkyl, or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive.

45. The compound of any one of the preceding claims, wherein R.sup.B is hydrogen or C.sub.1-6 alkyl.

46. The compound of any one of the preceding claims, wherein R.sup.N is hydrogen or C.sub.1-6 alkyl.

47. The compound of any one of the preceding claims, wherein R is independently selected from hydrogen, C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, (CH.sub.2).sub.q-E, (CH.sub.2).sub.qO-E, and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive.

48. The compound of any one of the preceding claims, wherein R.sup.1 is independently selected from hydrogen, C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, (CH.sub.2).sub.q-E, (CH.sub.2).sub.qO-E, and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive.

49. The compound of any one of the preceding claims, wherein at least one of R and R.sup.1 is selected from hydrogen, C.sub.1-6 alkyl, (CH.sub.2).sub.q-E, and (CH.sub.2).sub.qO-E.

50. The compound of any one of the preceding claims, wherein at least one of R and R.sup.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; and q is 0, 1, 2, 3, or 4.

51. The compound of any one of the preceding claims wherein E is phenyl, naphthyl, or pyridyl ring, with each phenyl, naphthyl, or pyridyl ring substituted by 0, 1, 2, or 3 substituents independently selected from halogen, OH, CN, NO.sub.2, NH.sub.2, NH(C.sub.1-C.sub.6 alkyl), N(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, and CF.sub.3.

52. The compound of any one of the preceding claims, wherein at least one of R and R.sup.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 0, 1, 2, 3, or 4; and E is phenyl or a cyclic sugar.

53. The compound of any one of the preceding claims, wherein at least one of R and R.sup.1 is a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive.

54. The compound of any one of the preceding claims, wherein each of R and R.sup.1 are polyhydroxylated alkyl groups having from 3 to 8 carbon atoms, inclusive.

55. The compound of any one of the preceding claims, wherein p is 0 or 1.

56. The compound of any one of the preceding claims, wherein R.sup.4 is halogen or C.sub.1-6 alkyl.

57. The compound of any one of the preceding claims, wherein X is a bond or O.

58. The compound of any one of the preceding claims, wherein Y.sup.2 and L.sup.4 are each independently bonds.

59. The compound of any one of the preceding claims, wherein each instance of Y.sup.1 and Y.sup.2 is independently selected from bond, CH.sub.2, O, NR.sup.8, OC(O), C(O)O, NR.sup.8C(O), and C(O)NR.sup.8.

60. The compound of any one of the preceding claims, wherein Y.sup.1 is of the formula: ##STR00572##

61. The compound of any one of the preceding claims, wherein each polyhydroxylated alkyl group is independently of one of the following formulae: ##STR00573##

62. The compound of any one of the preceding claims, wherein the polyhydroxylated alkyl group of any one of the preceding claims is of the formula: ##STR00574##

63. The compound of any one of the preceding claims, wherein the polyhydroxylated alkyl group of any one of the preceding claims is of the formula: ##STR00575##

64. The compound of any one of the preceding claims, wherein the polyhydroxylated alkyl group of any one of the preceding claims is of the formula: ##STR00576##

65. The compound of claim 1, wherein the compound is a compound of Table 1, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof.

66. The compound of claim 1, wherein the compound is of the formula: ##STR00577## or pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, wherein: D is C(O)O, OC(O), C(O)NH, or NHC(O); n=1-3; R is H or ##STR00578## and R.sup.1 is H or ##STR00579##

67. The compound of claim 1, wherein the compound is of the formula: ##STR00580## or pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, wherein: R.sup.2 is C(O)OR.sup.9 or tetrazolyl; R.sup.9 is hydrogen, methyl, or ethyl; R is H or ##STR00581## and R.sup.1 is H or ##STR00582##

68. A pharmaceutical composition comprising a compound of any one of claims 1-67, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, and a pharmaceutically acceptable carrier or excipient.

69. A pharmaceutical composition comprising a compound of any one of claims 1-67, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, and an osmolyte.

70. The pharmaceutical composition of claim 69, wherein the osmolyte is hypertonic saline.

71. The pharmaceutical composition of claim 69, wherein the osmolyte is a reduced sugar or ionic sugar.

72. The pharmaceutical composition of any one of claims 68-71, wherein the composition further comprises an excipient, wherein the excipient is a cyclodextrin.

73. The pharmaceutical composition of any one of claims 68-72, wherein the composition is suitable for inhalation.

74. The pharmaceutical composition of any one of claims 68-72, wherein the composition is a solution for aerosolization and administration by nebulizer.

75. The pharmaceutical composition of any one of claims 68-72, wherein the composition is suitable for administration by metered dose inhaler.

76. The pharmaceutical composition of any one of claims 68-69 and 71-72, wherein the composition is a dry powder for administration by dry powder inhaler.

77. The pharmaceutical composition of any one of claims 68-76 further comprising a therapeutically active agent selected from anti-inflammatory agents, anticholinergic agents, -agonists, CFTR modulators, P2Y2 receptor agonists, PY214 antagonist, peroxisome proliferator-activated receptor agonists, kinase inhibitors, mucoactive agents, hydrating agents, immune-modulatory agents, antiinfective agents, and antihistamines.

78. A method for blocking sodium channels in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-67, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, or pharmaceutical composition according to any one of claims 68-77.

79. A method for promoting hydration of mucosal surfaces, improving mucociliary clearance, or restoring mucosal defense in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-67, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, or pharmaceutical composition according to any one of claims 68-77.

80. A method for treating or preventing a disease or disorder in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-67, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, or pharmaceutical composition according to any one of claims 68-77.

81. The method of claim 80, wherein the disease or disorder is reversible or irreversible airway obstruction, chronic obstructive pulmonary disease (COPD), asthma, primary ciliary dyskinesia, bronchiectasis, bronchiectasis due to conditions other than cystic fibrosis, acute bronchitis, chronic bronchitis, post-viral cough, cystic fibrosis, idiopathic pulmonary fibrosis, pneumonia, panbronchiolitis, transplant-associate bronchiolitis, or ventilator-associated tracheobronchitis.

82. The method of claim 80, wherein the disease or disorder is dry mouth (xerostomia), dry skin, vaginal dryness, sinusitis, rhinosinusitis, nasal dehydration, nasal dehydration brought on by administering dry oxygen, dry eye, Sjogren's disease, otitis media, distal intestinal obstruction syndrome, esophagitis, constipation, mucus accumulation and inflammation, chronic diverticulitis, or fibrosis resulting from inflammation and oxidant stress in the airways driven by mucus accumulation.

83. A method for preventing ventilator-associated pneumonia in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-67, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, or pharmaceutical composition according to any one of claims 68-77.

84. A method for promoting ocular or corneal hydration in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-67, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, or pharmaceutical composition according to any one of claims 68-77.

85. A method for preventing, mitigating, and/or treating deterministic health effects to the respiratory tract and/or other bodily organs caused by respirable aerosols containing radionuclides in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-67, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, or pharmaceutical composition according to any one of claims 68-77.

86. The compound according to any one of claims 1-67, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, or pharmaceutical composition according to any one of claims 68-77, for use in treating or preventing a disease or disorder in a subject.

87. Use of a compound according to any one of claims 1-67, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled derivative thereof, or pharmaceutical composition according to any one of claims 68-77, for the preparation of a medicament.

Description

DETAILED DESCRIPTION

[0114] The present disclosure provides epithelial sodium channel (ENaC) inhibitors including compounds of Formula (I), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof. The disclosure also provides pharmaceutical compositions, combination therapies, kits, uses, methods of use, and methods of preparation of the compounds disclosed herein. In some embodiments, the compounds disclosed herein are advantageous over known ENaC inhibitors. In certain embodiments, the compounds, pharmaceutical compositions, combination therapies, kits, uses, and methods of use disclosed herein the exhibit an improved therapeutic index, including improved systemic safety (e.g., very limited secondary renal pharmacology), improve primary pharmacology (e.g., lower non-specific mucus binding), and/or improved target engagement and pulmonary retention (e.g., improved PK/PD profile) over those known in the art. Further, in certain embodiments, the compounds, pharmaceutical compositions, combination therapies, kits, uses, and methods of use disclosed herein the exhibit high plasma protein binding which may lower renal clearance.

Compounds

[0115] In one aspect, provided herein are compounds of Formula (I):

##STR00007##

and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, wherein: R.sup.W1 is hydrogen, halogen, optionally substituted alkyl, or N(R.sup.N).sub.2; [0116] R.sup.W2 is hydrogen, halogen, optionally substituted alkyl, or N(R.sup.N).sub.2; [0117] optionally where R.sup.W1 and R.sup.W2 are joined together to form optionally substituted heterocyclyl or optionally substituted heteroaryl; [0118] each instance of R.sup.N is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a nitrogen protecting group, or optionally two instances of R.sup.N bonded to the same nitrogen atom are joined together to form optionally substituted heterocyclyl or optionally substituted heteroaryl; [0119] each instance of L.sup.1, L.sup.2, L.sup.3, and L.sup.4 is independently a bond, optionally substituted C.sub.1-10 alkylene, optionally substituted C.sub.2-10 alkenylene, optionally substituted C.sub.2-10 alkynylene, optionally substituted C.sub.1-10 heteroalkylene, optionally substituted C.sub.2-10 heteroalkenylene, or optionally substituted C.sub.2-10 heteroalkynylene; [0120] each instance of R.sup.4 is independently halogen, CN, NO.sub.2, N.sub.3, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, OR.sup.O, N(R.sup.N).sub.2, or SR.sup.S; [0121] each instance of R.sup.O is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or an oxygen protecting group; [0122] each instance of R.sup.S is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a sulfur protecting group; [0123] each p is independently selected from 0, 1, 2, 3, and 4; [0124] each instance of Y.sup.1 and Y.sup.2 is independently a bond, CH.sub.2, O, S, NR.sup.8, C(O), S(O), S(O).sub.2, OC(O), OS(O).sub.2, C(O)O, S(O).sub.2O, NR.sup.8C(O), NR'S(O).sub.2, C(O)NR.sup.8, S(O).sub.2NR.sup.8, or

##STR00008## [0125] each instance of R.sup.8 is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a nitrogen protecting group, optionally wherein R.sup.8 is substituted with 0, 1, or 2 NRR.sup.1; [0126] Ring A is optionally substituted arylene, optionally substituted heteroarylene, optionally substituted heterocyclylene, or optionally substituted heteroarylene; [0127] Z is hydrogen, NRR.sup.1, N.sup.+(O.sup.)RR.sup.1, OR.sup.B, C(R.sup.1).sub.3, NR.sup.A(CO)R.sup.C, NR.sup.A(CO)OR.sup.B, NR.sup.A(CO)N(R.sup.A).sub.2, NR.sup.A(CNR.sup.A)N(R.sup.A).sub.2, (CO)OR.sup.B, (CO)N(R.sup.A).sub.2, or B; [0128] each instance of R.sup.A is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a nitrogen protecting group, optionally wherein two R.sup.A bonded to the name nitrogen atom are joined together to form optionally substituted heteroaryl or optionally substituted heterocyclyl; [0129] each instance of R.sup.B is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or an oxygen protecting group; [0130] each instance of R.sup.C is independently hydrogen or optionally substituted alkyl; [0131] each instance of R and R.sup.1 is independently hydrogen, optionally substituted alkyl, optionally substituted heteroalkyl, a polyhydroxylated alkyl group, a polyhydroxylated heteroalkyl group, optionally substituted acyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted -alkyl-E, optionally substituted -heteroalkyl-E, or a nitrogen protecting group, optionally wherein R and R.sup.1 bonded to the same nitrogen atom are joined together with the intervening atoms to form optionally substituted heterocyclyl or optionally substituted heteroaryl; [0132] each instance of E is independently optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl, optionally wherein E is a cyclic sugar; and [0133] B is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl, optionally wherein B is substituted with 0, 1, or 2 R.sup.1.

[0134] In some embodiments, the compound of Formula (I) is a compound of Formula (II):

##STR00009##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0135] In some embodiments, the compound of Formula (I) is of Formula (III):

##STR00010##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, wherein: c is selected from 0, 1, 2, 3, 4, 5, and 6; [0136] X is selected from a bond, CH.sub.2, O, N(R.sup.N) and S; [0137] each n is independently selected from 0, 1, 2, 3, 4, 5, and 6; [0138] each instance of R.sup.2 and R.sup.3 is independently selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, N(R.sup.9).sub.2, OR.sup.9, C(O)OR.sup.9, C(O)N(R.sup.9).sub.2, NR.sup.AC(O)R.sup.9, NR.sup.AC(O)OR.sup.9, NR.sup.AC(O)N(R.sup.9).sub.2, OC(O)R.sup.9, OC(O)OR.sup.9, OC(O)N(R.sup.9).sub.2, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heterocyclyl, and optionally substituted heteroaryl, optionally wherein R.sup.2 and R.sup.3 is substituted with 0, 1, or 2 N(R.sup.A).sub.2, C(O)OR.sup.B, and NR.sup.AC(O)R.sup.10; [0139] each instance of R.sup.9 is independently selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, an amino acid, a peptide comprising 2, 3, 4, 5, or 6 amino acids, or a nitrogen or oxygen protecting group, optionally wherein two R.sup.9 bonded to the same nitrogen atom are joined together to form optionally substituted heteroaryl or optionally substituted heterocyclyl; and optionally wherein R.sup.9 is substituted with 0, 1, or 2 groups selected from N(R.sup.A).sub.2, C(O)OR.sup.10, and NR.sup.AC(O)R.sup.10; [0140] each instance of R.sup.10 is independently selected from hydrogen or optionally substituted alkyl substituted with 0, 1, or 2 groups selected from N(R.sup.A).sub.2, C(O)OR.sup.B, and NR.sup.AC(O)R.sup.C; [0141] u is selected from 0 and 1; and [0142] t is selected from 0 and 1.

[0143] In certain embodiments, the compound of Formula (I) is of Formula (IV):

##STR00011##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0144] In some embodiments, the compound of Formula (I) is of Formula (V):

##STR00012##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0145] In certain embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00013##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, wherein D is O or NR.sup.8.

[0146] In some embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00014##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, wherein D is O or NR.sup.8.

[0147] In some embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00015##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0148] In some embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00016##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0149] In some embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00017##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, wherein D is O or NR.sup.8.

[0150] In some embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00018##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0151] In some embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00019##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0152] In some embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00020##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, wherein D is O or NR.sup.8.

[0153] In some embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00021##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0154] In some embodiments, the compound of Formula (I) is of one of the following formulae:

##STR00022##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0155] In certain embodiments, the compound of Formula (I) is of the formula:

##STR00023##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, wherein g is 0, 1, 2, 3, 4, or 5.

[0156] In some embodiments, the compound of Formula (I) is of the formula:

##STR00024##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0157] In some embodiments, the compound of Formula (I) is of the formula:

##STR00025##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0158] In certain embodiments, the compound of Formula (I) is of the formula:

##STR00026##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0159] In certain embodiments, the compound of Formula (I) is of the formula:

##STR00027##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0160] In some embodiments, the compound of Formula (I) is of the formula:

##STR00028##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0161] In certain embodiments, the compound of Formula (I) is of the formula:

##STR00029##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0162] In some embodiments, the compound of Formula (I) is of the formula:

##STR00030##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0163] In certain embodiments, the compound of Formula (I) is of the formula:

##STR00031##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0164] In some embodiments, the compound of Formula (I) is of the formula:

##STR00032##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0165] In certain embodiments, the compound of Formula (I) is of the formula:

##STR00033##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0166] In some embodiments, the compound of Formula (I) is of the formula:

##STR00034##

or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0167] In any of the formulae provided herein (e.g., Formula (I) and subgenera thereof), in certain embodiments, the group represented by the formula:

##STR00035##

is of the formula:

##STR00036##

In certain embodiments, the group represented by the formula:

##STR00037##

is of the formula:

##STR00038##

In certain embodiments, the group represented by the formula:

##STR00039##

is of the formula:

##STR00040##

In certain embodiments, the group represented by the formula:

##STR00041##

is of the formula:

##STR00042##

In certain embodiments, the group represented by the formula:

##STR00043##

is of the formula:

##STR00044##

In certain embodiments, the group represented by the formula:

##STR00045##

is of the formula:

##STR00046##

in certain embodiments, the group represented by the formula:

##STR00047##

is of the formula:

##STR00048##

[0168] In some embodiments, R.sup.W1 is hydrogen, halogen, optionally substituted alkyl, or N(R.sup.N).sub.2. In some embodiments, R.sup.W1 is hydrogen. In some embodiments, R.sup.W1 is halogen. In some embodiments, R.sup.W1 is chloro. In some embodiments, R.sup.W1 is bromo. In some embodiments, R.sup.W1 is iodo. In some embodiments, R.sup.W1 is fluoro. In some embodiments, R.sup.W1 is C.sub.1-6 alkyl. In some embodiments, R.sup.W1 is-N(R.sup.N).sub.2. In some embodiments, R.sup.W1 is NH.sub.2.

[0169] In some embodiments, R.sup.W2 is hydrogen, halogen, optionally substituted alkyl, or N(R.sup.N).sub.2. In some embodiments, R.sup.W2 is hydrogen. In some embodiments, R.sup.W2 is halogen. In some embodiments, R.sup.W2 is chloro. In some embodiments, R.sup.W2 is bromo. In some embodiments, R.sup.W2 is iodo. In some embodiments, R.sup.W2 is fluoro. In some embodiments, R.sup.W2 is C.sub.1-6 alkyl. In some embodiments, R.sup.W2 is N(R.sup.N).sub.2. In some embodiments, R.sup.W2 is NH.sub.2.

[0170] In some embodiments, R.sup.W1 and R.sup.W2 are joined together to form optionally substituted heterocyclyl or optionally substituted heteroaryl. In some embodiments, R.sup.W1 and R.sup.W2 are joined together to form optionally substituted heterocyclyl. In some embodiments, R.sup.W1 and R.sup.W2 are joined together to form optionally substituted heteroaryl. In some embodiments, the group represented by the formula:

##STR00049##

is of the formula:

##STR00050##

In some embodiments, the group represented by the formula:

##STR00051##

is of the formula:

##STR00052##

In some embodiments, the group represented by the formula:

##STR00053##

is of the formula:

##STR00054##

In some embodiments, the group represented by the formula:

##STR00055##

is of the formula:

##STR00056##

[0171] In some embodiments, R.sup.W1 is chloro and R.sup.W2 is NH.sub.2.

[0172] In certain embodiments, each instance of R.sup.N is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a nitrogen protecting group, or optionally two instances of R.sup.N bonded to the same nitrogen atom are joined together to form optionally substituted heterocyclyl or optionally substituted heteroaryl. In some embodiments, each instance of R.sup.N is the same. In some embodiments, each instance of R.sup.N is different. In certain embodiments, some instances of R.sup.N are the same and some instances of R.sup.N are different. In some embodiments, R.sup.N is hydrogen or C.sub.1-6 alkyl. In some embodiments, each instance of R.sup.N is hydrogen. In some embodiments, some instances R.sup.N are hydrogen. In some embodiments, each instance of R.sup.N is C.sub.1-6 alkyl. In certain embodiments, some instances of R.sup.N are C.sub.1-6 alkyl. In certain embodiments, some instances of R.sup.N are hydrogen, and some are C.sub.1-6 alkyl. In certain embodiments, some instances of R.sup.N are hydrogen, and some are methyl. In certain embodiments, two instances of R.sup.N bonded to the same nitrogen atom are each hydrogen. In certain embodiments, two instances of R.sup.N bonded to the same nitrogen atom are methyl. In certain embodiments, two instances of R.sup.N bonded to the same nitrogen atom are each independently hydrogen and methyl. In some embodiments, two instances of R.sup.N bonded to the same nitrogen atom are joined together to form optionally substituted heterocyclyl. In some embodiments, two instances of R.sup.N bonded to the same nitrogen atom are joined together to form optionally substituted heteroaryl. In some embodiments, R.sup.N is a nitrogen protecting group.

[0173] In some embodiments, each instance of L.sup.1, L.sup.2, L.sup.3, and L.sup.4 is independently a bond, optionally substituted C.sub.1-10 alkylene, optionally substituted C.sub.2-10 alkenylene, optionally substituted C.sub.2-10 alkynylene, optionally substituted C.sub.1-10heteroalkylene, optionally substituted C.sub.2-10heteroalkenylene, or optionally substituted C.sub.2-10 heteroalkynylene. In some embodiments, L.sup.1, L.sup.2, L.sup.3, and L.sup.4 are different. In certain embodiments, some of L.sup.1, L.sup.2, L.sup.3, and L.sup.4 are different.

[0174] In some embodiments, L.sup.1 is an optionally substituted C.sub.1-10 alkylene. In some embodiments, L.sup.1 is C.sub.1 alkylene. In some embodiments, L.sup.1 is C.sub.2 alkylene. In some embodiments, L.sup.1 is C.sub.3 alkylene. In some embodiments, L.sup.1 is C.sub.4 alkylene. In some embodiments, L.sup.1 is C.sub.5 alkylene. In certain embodiments, L.sup.1 is optionally substituted C.sub.1-10 heteroalkylene. In some embodiments, L.sup.1 is a bond.

[0175] In some embodiments, L.sup.2 is an optionally substituted C.sub.1-10 alkylene. In some embodiments, L.sup.2 is C.sub.1 alkylene. In some embodiments, L.sup.2 is C.sub.2 alkylene. In some embodiments, L.sup.2 is C.sub.3 alkylene. In some embodiments, L.sup.2 is C.sub.4 alkylene. In some embodiments, L.sup.2 is C.sub.5 alkylene. In certain embodiments, L.sup.2 is optionally substituted C.sub.1-10heteroalkylene. In certain embodiments, L.sup.2 is C.sub.1-10heteroalkylene, comprising 1-5 oxygen atoms. In certain embodiments, L.sup.2 is C.sub.1-10heteroalkylene, comprising 1 oxygen. In some embodiments, L.sup.2 is OCH.sub.2. In some embodiments, L.sup.2 is O(CH.sub.2).sub.2. In some embodiments, L.sup.2 is O(CH.sub.2).sub.3. In some embodiments, L.sup.2 is O(CH.sub.2).sub.4. In some embodiments, L.sup.2 is O(CH.sub.2).sub.5. In some embodiments, L.sup.2 is a bond.

[0176] In some embodiments, L.sup.3 is an optionally substituted C.sub.1-10 alkylene. In some embodiments, L.sup.3 is C.sub.1 alkylene. In some embodiments, L.sup.3 is C.sub.2 alkylene. In some embodiments, L.sup.3 is C.sub.3 alkylene. In some embodiments, L.sup.3 is C.sub.4 alkylene. In some embodiments, L.sup.3 is C.sub.5 alkylene. In some embodiments, L.sup.3 is C.sub.6 alkylene. In certain embodiments, L.sup.3 is optionally substituted C.sub.1-10heteroalkylene. In some embodiments, L.sup.3 is a bond.

[0177] In some embodiments, L.sup.4 is an optionally substituted C.sub.1-10 alkylene. In some embodiments, L.sup.4 is C.sub.1 alkylene. In some embodiments, L.sup.4 is C.sub.2 alkylene. In some embodiments, L.sup.4 is C.sub.3 alkylene. In some embodiments, L.sup.4 is C.sub.4 alkylene. In some embodiments, L.sup.4 is C.sub.5 alkylene. In some embodiments, L.sup.4 is C.sub.6 alkylene. In certain embodiments, L.sup.4 is optionally substituted C.sub.1-10heteroalkylene. In some embodiments, L.sup.4 is a bond.

[0178] In certain embodiments, each instance of L.sup.2, L.sup.3, and L.sup.4 is independently substituted with one or more of R.sup.2 or R.sup.3. In certain embodiments, L.sup.3 is not substituted with R.sup.2. In certain embodiments, L.sup.3 is substituted with one R.sup.2. In certain embodiments, L.sup.4 is not substituted with R.sup.3. In certain embodiments, L.sup.3 is substituted with one R.sup.3. Each instance of R.sup.2 and R.sup.3 is independently as defined herein.

[0179] In certain embodiments, each p is independently selected from 0, 1, 2, 3, and 4. In some embodiments, each instance of p is the same. In some embodiments, each instance of p is different. In certain embodiments, p is 0 or 1. In some embodiments, each p is 0. In some embodiments, one instance of p is 1 and the other instance of p is 0. In some embodiments, each p is 1. In certain embodiments, one instance of p is 0 and the other instance of p is 2.

[0180] In some embodiments, each instance of R.sup.4 is independently halogen, CN, NO.sub.2, N.sub.3, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted acyl, OR.sup.O, N(R.sup.N).sub.2, or SR.sup.S. In some embodiments, each instance of R.sup.4 is the same. In some embodiments, each instance of R.sup.4 is different. In certain embodiments, some instances of R.sup.4 are the same and some instances of R.sup.4 are different. In some embodiments, R.sup.4 is halogen or C.sub.1-6 alkyl. In some embodiments, R.sup.4 is halogen. In certain embodiments, R.sup.4 is chloro. In some embodiments, R.sup.4 is iodo. In some embodiments, R.sup.4 is bromo. In some embodiments, R.sup.4 is fluoro. In some embodiments, R.sup.4 is C.sub.1-6 alkyl. In some embodiments, R.sup.4 is methyl. In certain embodiments, R.sup.4 is NH.sub.2. In some embodiments, R.sup.4 is OH.

[0181] In some embodiments, one instance of p is 0 and one instance of p is 1, wherein R.sup.4 is alkyl. In some embodiments, one instance of p is 0 and one instance of p is 1, wherein R.sup.4 is methyl. In some embodiments, one instance of p is 0 and one instance of p is 1, wherein R.sup.4 is halogen. In some embodiments, one instance of p is 0 and one instance of p is 1, wherein R.sup.4 is chloro.

[0182] In some embodiments, each instance of R.sup.O is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or an oxygen protecting group. In some embodiments, each instance of R.sup.O is the same. In some embodiments, each instance of R.sup.O is different. In certain embodiments, some instances of R.sup.O are the same and some instances of R.sup.O are different. In some embodiments, R.sup.O is hydrogen. In some embodiments, R.sup.O is alkyl. In certain embodiments, R.sup.O is methyl. In some embodiments, R.sup.O is acyl. In certain embodiments, R.sup.O is an oxygen protecting group.

[0183] In certain embodiments, each instance of R.sup.S is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a sulfur protecting group. In some embodiments, each instance of R.sup.S is the same. In some embodiments, each instance of R.sup.S is different. In certain embodiments, some instances of R.sup.S are the same and some instances of R.sup.S are different. In some embodiments, R.sup.S is hydrogen. In certain embodiments, R.sup.S is alkyl. In some embodiments, R.sup.S is acyl. In certain embodiments, R.sup.S is a sulfur protecting group.

[0184] In some embodiments, each instance of Y.sup.1 and Y.sup.2 is independently a bond, CH.sub.2, O, S, NR.sup.8, C(O), S(O), S(O).sub.2, OC(O), OS(O).sub.2, C(O)O, S(O).sub.2O, NR.sup.8C(O), NR.sup.8S(O).sub.2, C(O)NR.sup.8, S(O).sub.2NR.sup.B, or

##STR00057##

In certain embodiments, each instance of Y.sup.1 and Y.sup.2 is independently selected from bond, CH.sub.2, O, NR.sup.8, OC(O), C(O)O, NR.sup.8C(O), and C(O)NR.sup.8. In some embodiments, Y.sup.1 and Y.sup.2 are the same. In some embodiments, Y.sup.1 and Y.sup.2 are different.

[0185] In some embodiments, Y.sup.1 is a bond. In some embodiments, Y.sup.1 is a OC(O). In certain embodiments, Y.sup.1 is C(O)O. In some embodiments, Y.sup.1 is NR.sup.8C(O). In certain embodiments, Y.sup.1 is C(O)NR.sup.8. In some embodiments, Y.sup.1 is NHC(O). In certain embodiments, Y.sup.1 is C(O)NH. In some embodiments, Y.sup.1 is NMeC(O). In certain embodiments, Y.sup.1 is C(O)NMe-. In certain embodiments, Y.sup.1 is

##STR00058##

[0186] In some embodiments, Y.sup.2 is a bond. In some embodiments, Y.sup.2 is a OC(O). In certain embodiments, Y.sup.2 is C(O)O. In some embodiments, Y.sup.2 is NR.sup.8C(O). In certain embodiments, Y.sup.2 is C(O)NR.sup.8. In some embodiments, Y.sup.2 is NHC(O). In certain embodiments, Y.sup.2 is C(O)NH. In some embodiments, Y.sup.2 is NMeC(O). In certain embodiments, Y.sup.2 is C(O)NMe-. In certain embodiments, Y.sup.2 is NR.sup.8. In certain embodiments, Y.sup.2 is NH. In certain embodiments, Y.sup.2 is NMe-.

[0187] In some embodiments, Y.sup.1 is O, S, NR.sup.8, C(O), S(O), S(O).sub.2, OC(O), OS(O).sub.2, C(O)O, S(O).sub.2O, NR.sup.8C(O), NR.sup.8S(O).sub.2, C(O)NR.sup.8, S(O).sub.2NR.sup.8, or

##STR00059##

and Y.sup.2 is a bond. In certain embodiments, Y.sup.1 is C(O)NR.sup.8 and Y.sup.2 is C(O)NR.sup.8. In some embodiments, Y.sup.1 is NR.sup.8C(O) and Y.sup.2 is NR.sup.8. In certain embodiments, Y.sup.1 is C(O)NH and Y.sup.2 is C(O)NH. In some embodiments, Y.sup.1 is NHC(O) and Y.sup.2 is NH.

[0188] In certain embodiments, Y.sup.2 is a bond and L.sup.4 is a bond.

[0189] In some embodiments, each instance of R.sup.8 is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a nitrogen protecting group, optionally wherein R.sup.8 is substituted with 0, 1, or 2 NRR.sup.1. In some embodiments, each instance of R.sup.8 is the same. In some embodiments, each instance of R.sup.8 is different. In certain embodiments, some instances of R.sup.8 are the same and some instances of R.sup.8 are different. In some embodiments, R.sup.8 is hydrogen or optionally substituted C.sub.1-6 alkyl. In some embodiments, R.sup.8 is hydrogen. In certain embodiments, R.sup.8 is optionally substituted alkyl. In some embodiments, R.sup.8 is C.sub.1-6 alkyl. In certain embodiments, R.sup.8 is methyl. In some embodiments, R.sup.8 is ethyl. In certain embodiments, R.sup.8 is propyl. In certain embodiments, R.sup.8 is butyl. In some embodiments, R.sup.8 is alkyl substituted with NRR.sup.1. In some embodiments, R.sup.8 is C.sub.1-6 substituted with NRR.sup.1. In some embodiments, R.sup.8 is C.sub.1 substituted with NRR.sup.1. In some embodiments, R.sup.8 is C.sub.2 substituted with NRR.sup.1. In some embodiments, R.sup.8 is C.sub.3 substituted with NRR.sup.1. In some embodiments, R.sup.8 is alkyl substituted with NRR.sup.1, wherein R and R.sup.1 are each hydrogen. In some embodiments, R.sup.8 is alkyl substituted with NRR.sup.1, wherein R is a polyhydroxylated alkyl group and R.sup.1 is hydrogen. In some embodiments, R.sup.8 is alkyl substituted with NRR.sup.1, wherein R and R.sup.1 are each a polyhydroxylated alkyl group.

[0190] In some embodiments, Ring A is optionally substituted arylene, optionally substituted heteroarylene, optionally substituted heterocyclylene, or optionally substituted heteroarylene. In some embodiments, Ring A is optionally substituted heterocyclylene. In some embodiments, optionally substituted 5- or 6-membered heterocyclylene. In certain embodiments, Ring A is piperdinylene or pyrrolidinylene. In some embodiments, Ring A is piperdinylene. In certain embodiments, Ring A is pyrrolidinylene.

[0191] In some embodiments, Ring A is selected from

##STR00060##

wherein: each R.sup.Y is independently hydrogen, halogen, O, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.O, N(R.sup.N).sub.2, SRS, optionally substituted acyl, C(O)OR.sup.O, or C(O)N(R.sup.N).sub.2; and y is selected from 0, 1, and 2.

[0192] In some embodiments, Ring A is

##STR00061##

In some embodiments, Ring A is

##STR00062##

In some embodiments, Ring A is

##STR00063##

In some embodiments, Ring A is

##STR00064##

In some embodiments, Ring A is

##STR00065##

[0193] In some embodiments, y is 0. In certain embodiments, y is 1. In certain embodiments, y is 2.

[0194] In some embodiments, each instance of R.sup.Y is the same. In some embodiments, each instance of R.sup.Y is different. In certain embodiments, some instances of R.sup.Y are the same and some instances of R.sup.Y are different. In certain embodiments, R.sup.Y is hydrogen. In certain embodiments, R.sup.Y is O. In some embodiments, R.sup.Y is C(O)OR.sup.O. In some embodiments, R.sup.Y is C(O)OH. In some embodiments, R.sup.Y is C(O)OCH.sub.3.

[0195] In some embodiments y is 1 and R.sup.Y is O, C(O)OH or C(O)OCH.sub.3.

[0196] In some embodiments, Ring A is

##STR00066##

y is 1; and R.sup.Y is C(O)OR.sup.O. In some embodiments, Ring A is

##STR00067##

y is 1; and R.sup.Y is C(O)OH or C(O)CH.sub.3.

##STR00068##

In some embodiments, Ring A is

##STR00069##

y is 1; R.sup.Y is O; and R.sup.N is hydrogen. In some embodiments, Ring A is

##STR00070##

y is 1; R.sup.Y is O; and RN is methyl.

[0197] In certain embodiments, Z is hydrogen, NRR.sup.1, N.sup.+(O.sup.)RR.sup.1, OR.sup.B, C(R.sup.1).sub.3, NR.sup.A(CO)R.sup.C, NR.sup.A(CO)ORB, NR.sup.A(CO)N(RA).sub.2, NR.sup.A(CNR.sup.A)N(R.sup.A).sub.2, (CO)OR.sup.B, (CO)N(R.sup.A).sub.2, or B. In some embodiments, Z is hydrogen, C(R.sup.1).sub.3, C(O)OR.sup.B, or NR.sup.A(CNR.sup.A)N(R.sup.A).sub.2. In some embodiments, Z is hydrogen.

[0198] In some embodiments, Z is C(R.sup.1).sub.3. In some embodiments, Z is C(alkyl).sub.3. In some embodiments, Z is CH(alkyl).sub.2. In some embodiments, Z is CH(C.sub.1-6 alkyl).sub.2. In some embodiments, Z is CH.sub.2(alkyl). In some embodiments, Z is CH.sub.2(C.sub.1-6 alkyl).

[0199] In some embodiments, Z is NRR.sup.1. In some embodiments, Z is NH.sub.2. In some embodiments, Z is NHMe. In some embodiments, Z is N(Me).sub.2. In certain embodiments, Z is NRR.sup.1 wherein R is hydrogen and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is NRR.sup.1 wherein R is optionally substituted alkyl and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is NRR.sup.1 wherein R is C.sub.1-6 alkyl and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is NRR.sup.1 wherein R is optionally substituted -alkyl-E and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is NRR.sup.1 wherein R is optionally substituted heteroalkyl and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is NRR.sup.1 wherein R is optionally substituted -heteroalkyl-E and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is NRR.sup.1 wherein R is (CH.sub.2).sub.q-E and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is NRR.sup.1 wherein R is (CH.sub.2).sub.qO-E and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is NRR.sup.1 wherein R is (CH.sub.2).sub.q-E; E is phenyl or a cyclic sugar; and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is NRR.sup.1 wherein R is (CH.sub.2).sub.qO-E; E is phenyl or a cyclic sugar; and R.sup.1 is a polyhydroxylated alkyl group.

[0200] In some embodiments, Z is

##STR00071##

In certain embodiments, Z is

##STR00072##

[0201] In some embodiments, Z is N.sup.+(O.sup.)RR.sup.1. In some embodiments, Z is N.sup.+(O.sup.)H.sub.2. In some embodiments, Z is N.sup.+(O.sup.)HMe. In some embodiments, Z is N.sup.+(O.sup.)(Me).sub.2. In certain embodiments, Z is N.sup.+(O.sup.)RR.sup.1 wherein R is hydrogen and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is N.sup.+(O.sup.)RR.sup.1 wherein R is optionally substituted alkyl and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is N.sup.+(O.sup.)RR.sup.1 wherein R is C.sub.1-6 alkyl and R.sup.1 is a polyhydroxylated alkyl group. In certain embodiments, Z is N.sup.+(O.sup.)RR.sup.1 wherein R and R.sup.1 are each independently polyhydroxylated alkyl groups.

[0202] In some embodiments, Z is

##STR00073##

In certain embodiments, Z is

##STR00074##

[0203] In some embodiments, Z is OR.sup.B. In some embodiments, Z is OH. In some embodiments, Z is OCH.sub.3. In some embodiments, Z is C(O)OR.sup.B. In some embodiments, Z is C(O)OH. In some embodiments, Z is C(O)OMe. In some embodiments, Z is C(O)O(C.sub.1-6 alkyl).

[0204] In some embodiments, Z is NR.sup.A(CNR.sup.A)N(RA).sub.2. In some embodiments, Z is NH(CNH)NH.sub.2.

[0205] In some embodiments, Z is B. In some embodiments, B is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl, optionally wherein B is substituted with 0, 1, or 2 R.sup.1. In some embodiments, B is optionally substituted phenyl, optionally substituted pyridinyl, optionally substituted piperidinyl, or optionally substituted indolyl. In some embodiments, B is phenyl, pyridinyl, piperidinyl, or indolyl with each phenyl, pyridinyl, piperidinyl, or indolyl ring substituted by 0, 1, 2, or 3 substituents independently selected from halogen, OR.sup.6, C.sub.1-6 alkyl substituted with 0 or 1 NRR.sup.1, and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms; wherein R.sup.6 is hydrogen, C.sub.1-6 alkyl substituted with 0 or 1 NRR.sup.1, or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms. In some embodiments, B is substituted with 1 or 2 groups of the formula:

##STR00075##

In some embodiments, B is phenyl. In some embodiments, B is substituted phenyl. In some embodiments, B is phenyl substituted with 0, 1, or 2 R.sup.1. In some embodiments, B is phenyl substituted with 1 or 2 substituents independently selected from OR.sup.6 and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms. In some embodiments, B is phenyl substituted with OH. In some embodiments, B is phenyl substituted with 1 or 2 polyhydroxylated alkyl group having from 3 to 8 carbon atoms. In some embodiments, B is phenyl substituted with 1 or 2 groups of the formula:

##STR00076##

In some embodiments, B is unsubstituted phenyl. In some embodiments, B is substituted indolyl. In some embodiments, B is indolyl substituted with 0, 1, or 2 R.sup.1. In some embodiments, B is indolyl substituted with 1 or 2 substituents independently selected from OR.sup.6 and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms. In some embodiments, B is unsubstituted indolyl. In some embodiments, B is substituted pyridinyl. In some embodiments, B is pyridinyl substituted with 0, 1, or 2 R.sup.1. In some embodiments, B is pyridinyl substituted with 1 or 2 substituents independently selected from OR.sup.6 and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms. In some embodiments, B is unsubstituted pyridinyl. In some embodiments, B is substituted piperidinyl. In some embodiments, B is piperidinyl substituted with 0, 1, or 2 R.sup.1. In some embodiments, B is piperidinyl substituted with 1 or 2 substituents independently selected from OR.sup.6 and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms. In some embodiments, B is unsubstituted piperidinyl. In some embodiments, B is substituted piperazinyl. In some embodiments, B is piperazinyl substituted with 0, 1, or 2 R.sup.1. In some embodiments, B is piperazinyl substituted with 1 or 2 substituents independently selected from OR.sup.6 and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms. In some embodiments, B is unsubstituted piperazinyl.

[0206] In some embodiments, each instance of R.sup.A is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or a nitrogen protecting group, optionally wherein two R.sup.A bonded to the name nitrogen atom are joined together to form optionally substituted heteroaryl or optionally substituted heterocyclyl. In some embodiments, each instance of R.sup.A is the same. In certain embodiments, each instance of R.sup.A is different. In some embodiments, some instances of R.sup.A are the same and some instances of R.sup.A are different. In certain embodiments, each R.sup.A is independently hydrogen, alkyl, or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive. In some embodiments, R.sup.A is hydrogen or alkyl. In some embodiments, R.sup.A is hydrogen or C.sub.1-6 alkyl. In some embodiments, R.sup.A is hydrogen. In some embodiments, R.sup.A is optionally substituted alkyl. In some embodiments, R.sup.A is optionally C.sub.1-6 substituted alkyl. In certain embodiments, R.sup.A is acyl (e.g., C(O)R.sup.O). In some embodiments, R.sup.A is a nitrogen protecting group. In certain embodiments, two R.sup.A bonded to the name nitrogen atom are joined together to form optionally substituted heterocyclyl. In some embodiments, two R.sup.A bonded to the name nitrogen atom are each hydrogen. In some embodiments, two R.sup.A bonded to the name nitrogen atom are each methyl. In some embodiments, two R.sup.A bonded to the name nitrogen atom are each independently hydrogen and methyl.

[0207] In some embodiments, each instance of R.sup.B is independently hydrogen, optionally substituted alkyl, optionally substituted acyl, or an oxygen protecting group. In some embodiments, each R.sup.B is independently hydrogen, C.sub.1-6 alkyl, or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive. In some embodiments, each instance of R.sup.B is the same. In some embodiments, each instance of R.sup.B is different. In certain embodiments, some instances of R.sup.B are the same and some instances of R.sup.B are different. In certain embodiments, R.sup.B is hydrogen or C.sub.1-6 alkyl. In certain embodiments, R.sup.B is hydrogen. In some embodiments, R.sup.B is optionally substituted alkyl. In some embodiments, R.sup.B is C.sub.1-6 alkyl. In some embodiments, R.sup.B is an oxygen protecting group.

[0208] In some embodiments, each instance of R.sup.C is independently hydrogen or optionally substituted alkyl. In some embodiments, each instance of R.sup.C is the same. In some embodiments, each instance of R.sup.C is different. In certain embodiments, some instances of R.sup.C are the same and some instances of R.sup.C are different. In some embodiments, R.sup.C is hydrogen. In certain embodiments, R.sup.C is optionally substituted alkyl. In some embodiments, R.sup.C is hydrogen. In certain embodiments, R.sup.C is substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.C is unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.C is methyl. In certain embodiments, R.sup.C is ethyl. In certain embodiments, R.sup.C is propyl. In certain embodiments, R.sup.C is butyl. In certain embodiments, R.sup.C is tert-butyl.

[0209] In certain embodiments, each instance of R and R.sup.1 is independently hydrogen, optionally substituted alkyl, optionally substituted heteroalkyl, a polyhydroxylated alkyl group, a polyhydroxylated heteroalkyl group, optionally substituted acyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted -alkyl-E, optionally substituted -heteroalkyl-E, or a nitrogen protecting group, optionally wherein R and R.sup.1 bonded to the same nitrogen atom are joined together with the intervening atoms to form optionally substituted heterocyclyl or optionally substituted heteroaryl. In certain embodiments, each instance of R and R.sup.1 are the same. In some embodiments, each instance of R and R.sup.1 are different. In some embodiments, some instances of R are the same and some instances of R are different. In certain embodiments, some instances of R.sup.1 are the same and some instances of R.sup.1 are different.

[0210] In some embodiments, R is selected from hydrogen, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted -alkyl-E, optionally substituted -heteroalkyl-E, and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive. In some embodiments, R is selected from hydrogen, C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, (CH.sub.2).sub.q-E, (CH.sub.2).sub.qO-E, and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive. In some embodiments, R is hydrogen. In some embodiments, R is optionally substituted alkyl. In some embodiments, R is optionally substituted C.sub.1-6 alkyl. In some embodiments, R is methyl. In certain embodiments, R is ethyl. In certain embodiments, R is ethyl substituted with OH. In some embodiments, R is propyl. In some embodiments, R is isopropyl. In some embodiments, R is butyl. In some embodiments, R is pentyl. In some embodiments, R is hexyl. In some embodiments, R is optionally substituted heteroalkyl. In some embodiments, R is -alkyl-E. In some embodiments, R is (CH.sub.2).sub.q-E. In some embodiments, R is -heteroalkyl-E. In some embodiments, R is (CH.sub.2).sub.qO-E. In some embodiments, R is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; and q is 0, 1, 2, 3, or 4. In some embodiments, R is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 0, 1, 2, 3, or 4; and E is phenyl or a cyclic sugar. In some embodiments, R is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is optionally substituted aryl. In some embodiments, R is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is phenyl. In some embodiments, R is (CH.sub.2).sub.q-E; q is 2 or 4; and E is phenyl. In some embodiments, R is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is optionally substituted heterocyclyl. In some embodiments, R is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is a cyclic sugar. In some embodiments, R is (CH.sub.2).sub.q-E; [0211] q is 2 or 4; and E is a cyclic sugar. In some embodiments, R is a polyhydroxylated alkyl group having from 3 to 8 carbon atoms. In some embodiments, R is of the formula:

##STR00077##

In some embodiments, R is of the formula:

##STR00078##

In some embodiments, R is of the formula:

##STR00079##

In some embodiments, R is of the formula:

##STR00080##

In some embodiments, R is of the formula:

##STR00081##

[0212] In some embodiments, R.sup.1 is selected from hydrogen, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted -alkyl-E, optionally substituted -heteroalkyl-E, and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive. In some embodiments, R.sup.1 is selected from hydrogen, C.sub.1-6 alkyl, C.sub.1-6 heteroalkyl, (CH.sub.2).sub.q-E, (CH.sub.2).sub.qO-E, and a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive. In some embodiments, R.sup.1 is hydrogen. In some embodiments, R.sup.1 is optionally substituted alkyl. In some embodiments, R.sup.1 is optionally substituted C.sub.1-6 alkyl. In some embodiments, R.sup.1 is methyl. In certain embodiments, R.sup.1 is ethyl. In certain embodiments, R.sup.1 is ethyl substituted with OH. In some embodiments, R.sup.1 is propyl. In some embodiments, R.sup.1 is isopropyl. In some embodiments, R.sup.1 is butyl. In some embodiments, R.sup.1 is pentyl. In some embodiments, R.sup.1 is hexyl. In some embodiments, R.sup.1 is optionally substituted heteroalkyl. In some embodiments, R.sup.1 is -alkyl-E. In some embodiments, R.sup.1 is (CH.sub.2).sub.q-E. In some embodiments, R.sup.1 is -heteroalkyl-E. In some embodiments, R.sup.1 is (CH.sub.2).sub.qO-E. In some embodiments, R.sup.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; and q is 0, 1, 2, 3, or 4. In some embodiments, R.sup.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 0, 1, 2, 3, or 4; and E is phenyl or a cyclic sugar. In some embodiments, R.sup.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is optionally substituted aryl. In some embodiments, R.sup.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is phenyl. In some embodiments, R.sup.1 is (CH.sub.2).sub.q-E; q is 2 or 4; and E is phenyl. In some embodiments, R.sup.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is optionally substituted heterocyclyl. In some embodiments, R.sup.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is a cyclic sugar. In some embodiments, R.sup.1 is (CH.sub.2).sub.q-E; q is 2 or 4; and E is a cyclic sugar. In some embodiments, R.sup.1 is a polyhydroxylated alkyl group having from 3 to 8 carbon atoms. In some embodiments, R.sup.1 is of the formula:

##STR00082##

In some embodiments, R.sup.1 is of the formula:

##STR00083##

In some embodiments, R.sup.1 is of the formula:

##STR00084##

In some embodiments, R.sup.1 is of the formula:

##STR00085##

In some embodiments, R.sup.1 is of the formula:

##STR00086##

[0213] In some embodiments, at least one of R and R.sup.1 is hydrogen. In certain embodiments, both of R and R.sup.1 are hydrogen. In some embodiments, at least one of R and R.sup.1 is methyl. In certain embodiments, both of R and R.sup.1 are methyl. In some embodiments, at least one of R and R.sup.1 is a polyhydroxylated alkyl groups having from 3 to 8 carbon atoms, inclusive. In certain embodiments, both of R and R.sup.1 are polyhydroxylated alkyl groups having from 3 to 8 carbon atoms, inclusive. In some embodiments, at least one of R and R.sup.1 is selected from hydrogen, C.sub.1-6 alkyl, (CH.sub.2).sub.q-E, and (CH.sub.2).sub.qO-E. In some embodiments, at least one of R and R.sup.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; and q is 0, 1, 2, 3, or 4. In some embodiments, at least one of R and R.sup.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 0, 1, 2, 3, or 4; and E is phenyl or a cyclic sugar. In some embodiments, each of R and R.sup.1 are polyhydroxylated alkyl groups having from 3 to 8 carbon atoms, inclusive. In some embodiments, at least one of R and R.sub.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is optionally substituted aryl. In some embodiments, at least one of R and R.sub.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is phenyl. In some embodiments, at least one of R and R.sub.1 is (CH.sub.2).sub.q-E; q is 2 or 4; and E is phenyl. In some embodiments, at least one of R and R.sub.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is optionally substituted heterocyclyl. In some embodiments, at least one of R and R.sub.1 is (CH.sub.2).sub.q-E or (CH.sub.2).sub.qO-E; q is 1, 2, 3, or 4; and E is a cyclic sugar. In some embodiments, at least one of R and R.sub.1 is (CH.sub.2).sub.q-E; q is 2 or 4; and E is a cyclic sugar.

[0214] In some embodiments, each instance of E is independently optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, or optionally substituted heterocyclyl, optionally wherein E is a cyclic sugar. In some embodiments, E is phenyl, naphthyl, or pyridyl ring, with each phenyl, naphthyl, or pyridyl ring substituted by 0, 1, 2, or 3 substituents independently selected from halogen, OH, CN, NO.sub.2, NH.sub.2, NH(C.sub.1-C.sub.6 alkyl), N(C.sub.1-C.sub.6 alkyl).sub.2, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, and CF.sub.3. In some embodiments, each instance of E is the same. In some embodiments, each instance of E is different. In certain embodiments, some instances of E are the same and some instances of E are different. In some embodiments, E is phenyl or a cyclic sugar. In some embodiments, E is optionally substituted aryl. In certain embodiments, E is phenyl. In some embodiments, E is optionally substituted heterocyclyl. In some embodiments, E is a cyclic sugar. In some embodiments, E is a cyclic sugar of the formula:

##STR00087##

In some embodiments, E is a cyclic sugar of the formula:

##STR00088##

In some embodiments, E is a cyclic sugar of the formula:

##STR00089##

[0215] In some embodiments, c is selected from 0, 1, 2, 3, 4, 5, and 6. In some embodiments, c is 0. In some embodiments, c is 1. In some embodiments, c is 2. In some embodiments, c is 3. In some embodiments, c is 4. In some embodiments, c is 5. In some embodiments, c is 6.

[0216] In some embodiments, X is selected from a bond, CH.sub.2, O, N(R.sup.N) and S. In some embodiments, X is a bond or O. In some embodiments, X is a bond. In some embodiments, X is CH.sub.2. In some embodiments, X is-N(R.sup.N). In some embodiments, X is NH. In some embodiments, X is N(CH.sub.3). In some embodiments, X is O. In some embodiments, X is S.

[0217] In some embodiments, each n is independently selected from 0, 1, 2, 3, 4, 5, and 6. In some embodiments, each n is the same. In some embodiments, each n is different. In some embodiments, some instances of n are the same and some instances of n are different. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, each instance of n is 2. In some embodiments, each instance of n is 3. In some embodiments, each instance of n is 4. In some embodiments, one instance of n is 2 and one instance of n is 4. In some embodiments, one instance of n is 3 and one instance of n is 1. In some embodiments, one instance of n is 2 and one instance of n is 3. In some embodiments, one instance of n is 0 and one instance of n is 2. In some embodiments, one instance of n is 1 and one instance of n is 2.

[0218] In some embodiments, R.sup.2 is independently selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, N(R.sup.9).sub.2, OR.sup.9, C(O)OR.sup.9, C(O)N(R.sup.9).sub.2, NR.sup.AC(O)R.sup.9, NR.sup.AC(O)OR.sup.9, NR.sup.AC(O)N(R.sup.9).sub.2, OC(O)R.sup.9, OC(O)OR.sup.9, OC(O)N(R.sup.9).sub.2, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heterocyclyl, and optionally substituted heteroaryl, optionally wherein R.sup.2 is substituted with 0, 1, or 2 N(R.sup.A).sub.2, C(O)OR.sup.B, and NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.2 is one of the following formulae:

##STR00090##

or salt thereof. In some embodiments, each instance of R.sup.2 is the same. In some embodiments, each instance of R.sup.2 is different. In certain embodiments, some instances of R.sup.2 are the same and some instances of R.sup.2 are different. In some embodiments, R.sup.2 is hydrogen. In some embodiments, R.sup.2 is N(R.sup.9).sub.2. In some embodiments, R.sup.2 is NH.sub.2. In some embodiments, R.sup.2 is NMe.sub.2. In some embodiments, R.sup.2 is a C.sub.1-6 alkyl substituted with N(R.sup.9).sub.2. In some embodiments, R.sup.2 is C(O)OR.sup.9. In some embodiments, R.sup.2 is C(O)OH. In some embodiments, R.sup.2 is C(O)ONa.sup.+. In some embodiments, R.sup.2 is C(O)OMe. In some embodiments, R.sup.2 is C(O)N(R.sup.9).sub.2. In some embodiments, R.sup.2 is C(O)NH.sub.2. In some embodiments, R.sup.2 is C(O)N(Me).sub.2. In some embodiments, R.sup.2 is NR.sup.AC(O)OR.sup.9. In some embodiments, R.sup.2 is NR.sup.AC(O)OH. In some embodiments, R.sup.2 is NHC(O)OH. In some embodiments, R.sup.2 is NR.sup.AC(O)OMe. In some embodiments, R.sup.2 is NHC(O)OMe. In some embodiments, R.sup.2 is NR.sup.AC(O)OtBu. In some embodiments, R.sup.2 is NHC(O)OtBu. In some embodiments, R.sup.2 is NR.sup.AC(O)R.sup.9. In some embodiments, R.sup.2 is NHC(O)R.sup.9. In some embodiments, R.sup.2 is NHC(O)(CH.sub.2).sub.1-6C(O)OH. In some embodiments, R.sup.2 is NHC(O)(CH.sub.2).sub.1-6C(O)OMe. In some embodiments, R.sup.2 is NHC(O)(CH.sub.2).sub.1-6C(O)OtBu. In some embodiments R.sup.2 is

##STR00091##

[0219] In some embodiments, R.sup.3 is selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, N(R.sup.9).sub.2, OR.sup.9, C(O)OR.sup.9, C(O)N(R.sup.9).sub.2, NR.sup.AC(O)R.sup.9, NR.sup.AC(O)OR.sup.9, NR.sup.AC(O)N(R.sup.9).sub.2, OC(O)R.sup.9, OC(O)OR.sup.9, OC(O)N(R.sup.9).sub.2, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heterocyclyl, and optionally substituted heteroaryl, optionally wherein R.sup.3 is substituted with 0, 1, or 2 N(R.sup.A).sub.2, C(O)OR.sup.B, and NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.3 is one of the following formulae:

##STR00092##

or salt thereof. In some embodiments, R.sup.3 is hydrogen. In some embodiments, R.sup.3 is N(R.sup.9).sub.2. In some embodiments, R.sup.3 is NH.sub.2. In some embodiments, R.sup.3 is NMe.sub.2. In some embodiments, R.sup.3 is a C.sub.1-6 alkyl substituted with N(R.sup.9).sub.2. In some embodiments, R.sup.3 is C(O)OR.sup.9. In some embodiments, R.sup.3 is C(O)OH. In some embodiments, R.sup.3 is C(O)ONa.sup.+. In some embodiments, R.sup.3 is C(O)OMe. In some embodiments, R.sup.3 is C(O)N(R.sup.9).sub.2. In some embodiments, R.sup.2 is C(O)NH.sub.2. In some embodiments, R.sup.3 is C(O)N(Me).sub.2. In some embodiments, R.sup.3 is NR.sup.AC(O)OR.sup.9. In some embodiments, R.sup.3 is NR.sup.AC(O)OH. In some embodiments, R.sup.3 is NHC(O)OH. In some embodiments, R.sup.3 is NR.sup.AC(O)OMe. In some embodiments, R.sup.3 is NHC(O)OMe. In some embodiments, R.sup.3 is NR.sup.AC(O)OtBu. In some embodiments, R.sup.3 is NHC(O)OtBu. In some embodiments, R.sup.3 is NR.sup.AC(O)R.sup.9. In some embodiments, R.sup.3 is NHC(O)R.sup.9. In some embodiments, R.sup.3 is NHC(O)(CH.sub.2).sub.1-6C(O)OH. In some embodiments, R.sup.3 is NHC(O)(CH.sub.2).sub.1-6C(O)OMe. In some embodiments, R.sup.3 is NHC(O)(CH.sub.2).sub.1-6C(O)OtBu.

[0220] In some embodiments, R.sup.2 is hydrogen and R.sup.3 is hydrogen.

[0221] In some embodiments, each instance of R.sup.9 is independently selected from hydrogen, optionally substituted alkyl, optionally substituted acyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, an amino acid, a peptide comprising 2, 3, 4, 5, or 6 amino acids, or a nitrogen or oxygen protecting group, optionally wherein two R.sup.9 bonded to the same nitrogen atom are joined together to form optionally substituted heteroaryl or optionally substituted heterocyclyl; and optionally wherein R.sup.9 is substituted with 0, 1, or 2 groups selected from N(R.sup.A).sub.2, C(O)OR.sup.10, and NR.sup.AC(O)R.sup.10. In certain embodiments, at least one instance of R.sup.9 is hydrogen or a C.sub.1-6 alkyl substituted with 0, 1, or 2 groups selected from N(R.sup.A).sub.2, C(O)OR.sup.10, and NR.sup.AC(O)R.sup.10. In some embodiments, at least one instance of R.sup.9 is independently hydrogen, C.sub.1-6alkyl, or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive. In some embodiments, each instance of R.sup.9 is the same. In some embodiments, each instance of R.sup.9 is different. In certain embodiments, some instances of R.sup.9 are the same and some instances of R.sup.9 are different. In some embodiments, R.sup.9 is hydrogen. In some embodiments, R.sup.9 is alkyl. In some embodiments, R.sup.9 is methyl. In some embodiments, R.sup.9 is ethyl. In some embodiments, R.sup.9 is propyl. In some embodiments, R.sup.9 is butyl. In some embodiments, R.sup.9 is tert-butyl. In some embodiments, R.sup.9 is alkyl substituted with 0, 1, or 2 groups selected from N(R.sup.A).sub.2, C(O)OR.sup.10, and NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.9 is alkyl substituted with 2 independent N(R.sup.A).sub.2. In some embodiments, R.sup.9 is alkyl substituted with 2 NH.sub.2. In some embodiments, R.sup.9 is C.sub.1-6 alkyl substituted with 2 independent N(R.sup.A).sub.2. In some embodiments, R.sup.9 is C.sub.1-6 alkyl substituted with 2 NH.sub.2. In some embodiments, R.sup.9 is C.sub.5 alkyl substituted with 2 independent N(R.sup.A).sub.2. In some embodiments, R.sup.9 is C.sub.5 alkyl substituted with 2 NH.sub.2. In some embodiments, R.sup.9 is 1,5-amino-C.sub.5 alkyl. In some embodiments, R.sup.9 is alkyl substituted with N(R.sup.A).sub.2. In some embodiments, R.sup.9 is alkyl substituted with NH.sub.2. In some embodiments, R.sup.9 is alkyl substituted with NMe.sub.2. In some embodiments, R.sup.9 is alkyl substituted with NHMe. In some embodiments, R.sup.9 is C.sub.1-6 alkyl substituted with N(R.sup.A).sub.2. In some embodiments, R.sup.9 is C.sub.1-6 alkyl substituted with NH.sub.2. In some embodiments, R.sup.9 is C.sub.1-6 alkyl substituted with NMe.sub.2. In some embodiments, R.sup.9 is C.sub.1-6 alkyl substituted with NHMe. In some embodiments, R.sup.9 is alkyl substituted with C(O)OR.sup.10. In some embodiments, R.sup.9 is C.sub.1 alkyl substituted with C(O)OR.sup.10. In some embodiments, R.sup.9 is C.sub.2 alkyl substituted with C(O)OR.sup.10. In some embodiments, R.sup.9 is C.sub.3 alkyl substituted with C(O)OR.sup.10. In some embodiments, R.sup.9 is alkyl C.sub.4 substituted with C(O)OR.sup.10. In some embodiments, R.sup.9 is C.sub.5 alkyl substituted with C(O)OR.sup.10. In some embodiments, R.sup.9 is C.sub.6 alkyl substituted with C(O)OR.sup.10. In some embodiments, R.sup.9 is alkyl substituted with NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.1 alkyl substituted with NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.2 alkyl substituted with NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.3 alkyl substituted with NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.4 alkyl substituted with NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.5 alkyl substituted with NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.6 alkyl substituted with NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.9 is alkyl substituted with NR.sup.AC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.1 alkyl substituted with NHC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.2 alkyl substituted with NHC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.3 alkyl substituted with NHC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.4 alkyl substituted with NHC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.5 alkyl substituted with NHC(O)R.sup.10. In some embodiments, R.sup.9 is C.sub.6 alkyl substituted with NHC(O)R.sup.10.

[0222] In some embodiments, each instance of R.sup.10 is independently selected from hydrogen or optionally substituted alkyl substituted with 0, 1, or 2 groups selected from N(R.sup.A).sub.2, C(O)OR.sup.B, and NR.sup.AC(O)R.sup.C. In certain embodiments, R.sup.10 is independently hydrogen or C.sub.1-6 alkyl substituted with N(R.sup.A).sub.2, C(O)OR.sup.B, and NR.sup.AC(O)R.sup.C. In some embodiments, each instance of R.sup.10 is the same. In some embodiments, each instance of R.sup.10 is different. In certain embodiments, some instances of R.sup.10 are the same and some instances of R.sup.10 are different. In some embodiments, R.sup.10 is hydrogen. In some embodiments, R.sup.10 is alkyl. In some embodiments, R.sup.10 is alkyl substituted with C(O)OR.sup.B. In some embodiments, R.sup.10 is alkyl substituted with C(O)OH. In some embodiments, R.sup.10 is alkyl substituted with C(O)OMe. In some embodiments, R.sup.10 is alkyl substituted with C(O)OtBu. In some embodiments, R.sup.10 is alkyl substituted with NR.sup.AC(O)R.sup.C. In some embodiments, R.sup.10 is alkyl substituted with NHC(O)H. In some embodiments, R.sup.10 is alkyl substituted with NHC(O)Me. In some embodiments, R.sup.10 is alkyl substituted with NHC(O)tBu. In some embodiments, R.sup.10 is alkyl substituted with N(R.sup.A).sub.2. In some embodiments, R.sup.10 is alkyl substituted with N(R.sup.A).sub.2. In some embodiments, R.sup.10 is alkyl substituted with NH.sub.2. In some embodiments, R.sup.10 is C.sub.1 alkyl substituted with N(R.sup.A).sub.2. In some embodiments, R.sup.10 is C.sub.2 alkyl substituted with N(R.sup.A).sub.2. In some embodiments, R.sup.10 is C.sub.3 alkyl substituted with N(R.sup.A).sub.2. In some embodiments, R.sup.10 is C.sub.4 alkyl substituted with N(R.sup.A).sub.2. In some embodiments, R.sup.10 is C.sub.5 alkyl substituted with N(R.sup.A).sub.2. In some embodiments, R.sup.10 is C.sub.6 alkyl substituted with N(R.sup.A).sub.2. In some embodiments, R.sup.10 is C.sub.1 alkyl substituted with NH.sub.2. In some embodiments, R.sup.10 is C.sub.2 alkyl substituted with NH.sub.2. In some embodiments, R.sup.10 is C.sub.3 alkyl substituted with NH.sub.2. In some embodiments, R.sup.10 is C.sub.4 alkyl substituted with NH.sub.2. In some embodiments, R.sup.10 is C.sub.5 alkyl substituted with NH.sub.2. In some embodiments, R.sup.10 is C.sub.6 alkyl substituted with NH.sub.2.

[0223] In some embodiments, u is selected from 0 and 1. In some embodiments, u is 0. In certain embodiments, u is 1.

[0224] In some embodiments, t is selected from 0 and 1. In some embodiments t is 0. In certain embodiments, t is 1.

[0225] In some embodiments, D is O or NR.sup.8. In some embodiments, D is O. In some embodiments, D is NR.sup.8. In some embodiments, D is NH. In some embodiments, D is NMe-. In some embodiments, D is NR.sup.8, wherein R.sup.1 is C.sub.1-6 alkyl. In some embodiments, D is NR.sup.8, wherein R.sup.8 is C.sub.1-6 alkyl substituted with NRR.sup.1. In some embodiments, D is NR.sup.8, wherein R.sup.1 is C.sub.1-6 alkyl substituted with NH.sub.2. In some embodiments, D is NR.sup.8, wherein R.sup.1 is C.sub.1-6 alkyl substituted with NMe.sub.2. In some embodiments, D is NR.sup.8, wherein R.sup.1 is C.sub.1-6 alkyl substituted with NHMe. In some embodiments, D is NR.sup.8, wherein R.sup.1 is C.sub.1-6 alkyl substituted with NRR.sup.1, wherein R.sup.1 is hydrogen and R is a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive. In some embodiments, D is NR.sup.8, wherein R.sup.1 is C.sub.1-6 alkyl substituted with NRR.sup.1, wherein R and R.sup.1 are each independently a polyhydroxylated alkyl group having from 3 to 8 carbon atoms, inclusive.

[0226] In some embodiments, g is selected from 0, 1, 2, 3, 4, and 5. In some embodiments, g is 0. In some embodiments, g is 1. In some embodiments, g is 2. In some embodiments, g is 3. In some embodiments, g is 4. In some embodiments, g is 5.

[0227] In some embodiments, NRR.sup.1 is NH.sub.2. In some embodiments, NRR.sup.1 is NHCH.sub.3. In some embodiments, NRR.sup.1 is N(CH.sub.3).sub.2. In some embodiments, NRR.sup.1 is of the formula:

##STR00093##

In some embodiments, NRR.sup.1 is of the formula:

##STR00094##

In some embodiments, NRR.sup.1 is of the formula:

##STR00095##

In some embodiments, NRR.sup.1 is of the formula:

##STR00096##

In some embodiments, NRR.sup.1 is of the formula:

##STR00097##

[0228] In some embodiments, NRR.sup.1 is of the formula:

##STR00098##

In some embodiments, NRR.sup.1 is of the formula:

##STR00099##

In some embodiments, NRR.sup.1 is of the formula:

##STR00100##

[0229] In some embodiments, a polyhydroxylated alkyl group (e.g., a polyhydroxylated alkyl group having from 3 to 8 carbon atoms) is of the formula:

##STR00101##

In some embodiments, a polyhydroxylated alkyl group (e.g., a polyhydroxylated alkyl group having from 3 to 8 carbon atoms) is of the formula:

##STR00102##

In some embodiments, the polyhydroxylated alkyl group or polyhydroxylated alkyl group or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms is of the formula:

##STR00103##

In some embodiments, the polyhydroxylated alkyl group or polyhydroxylated alkyl group or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms is of the formula:

##STR00104##

In some embodiments, the polyhydroxylated alkyl group or polyhydroxylated alkyl group or a polyhydroxylated alkyl group having from 3 to 8 carbon atoms is of the formula:

##STR00105##

[0230] In some embodiments, the compound of Formula (I) is a compound of the formula:

##STR00106##

wherein D is C(O)O, OC(O), C(O)NH, or NHC(O); n=1-3; R is H or Sug; and R.sup.1 is H or Sug; wherein Sug is a sugar represented by the structure:

##STR00107##

[0231] In some embodiments, the compound of Formula (I) is a compound of the formula:

##STR00108##

wherein R.sup.2 is C(O)OR.sup.9 or tetrazolyl; R.sup.9 is hydrogen, methyl, or ethyl; R is H or Sug; and R.sup.1 is H or Sug; wherein Sug is a sugar represented by the structure:

##STR00109##

[0232] In some embodiments, a compound of Formula (I) is a compound selected from those listed in Table 1 (below), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof.

TABLE-US-00001 TABLE 1 Ex- am- ple Structure 1 [00110] 2 [00111] 3 [00112] 4 [00113] 5 [00114] 6 [00115] 7 [00116] 8 [00117] 9 [00118] 10 [00119] 11 [00120] 12 [00121] 13 [00122] 14 [00123] 15 [00124] 16 [00125] 17 [00126] 18 [00127] 19 [00128] 20 [00129] 21 [00130] 22 [00131] 23 [00132] 24 [00133] 25 [00134] 26 [00135] 27 [00136] 28 [00137] 29 [00138] 30 [00139] 31 [00140] 32 [00141] 33 [00142] 34 [00143] 35 [00144] 36 [00145] 37 [00146] 38 [00147] 39 [00148] 40 [00149] 41 [00150] 42 [00151] 43 [00152] 44 [00153] 45 [00154] 46 [00155] 47 [00156] 48 [00157] 49 [00158] 50 [00159] 51 [00160] 52 [00161] 53 [00162] 54 [00163] 55 [00164] 56 [00165] 57 [00166] 58 [00167] 59 [00168] 60 [00169] 61 [00170] 62 [00171] 63 [00172] 64 [00173] 65 [00174] 66 [00175] 67 [00176] 68 [00177] 69 [00178] 70 [00179] 71 [00180] 72 [00181] 73 [00182] 74 [00183] 75 [00184] 76 [00185] 77 [00186] 78 [00187] 79 [00188] 80 [00189] 81 [00190] 82 [00191] 83 [00192] 84 [00193] 85 [00194] 86 [00195] 87 [00196] 88 [00197] 89 [00198] 90 [00199] 91 [00200] 92 [00201] 93 [00202] 94 [00203] 95 [00204] 96 [00205] 97 [00206] 98 [00207] 99 [00208] 100 [00209] 101 [00210] 102 [00211] 103 [00212] 104 [00213] 105 [00214] 106 [00215] 107 [00216] 108 [00217] 109 [00218] 110 [00219] 111 [00220] 112 [00221] 113 [00222] 114 [00223] 115 [00224] 116 [00225] 117 [00226] 118 [00227] 119 [00228] 120 [00229] 121 [00230] 122 [00231] 123 [00232] 124 [00233] 125 [00234] 126 [00235] 127 [00236] 128 [00237] 129 [00238] 130 [00239] 131 [00240] 132 [00241] 133 [00242] 134 [00243] 135 [00244] 136 [00245] 137 [00246] 138 [00247] 139 [00248] 140 [00249] 141 [00250] 142 [00251] 143 [00252] 144 [00253] 145 [00254] 146 [00255] 147 [00256] 148 [00257] 149 [00258] 150 [00259] 151 [00260] 152 [00261] 153 [00262] 154 [00263] 155 [00264] 156 [00265] 157 [00266] 158 [00267] 159 [00268] 160 [00269] 161 [00270] 162 [00271] 163 [00272] 164 [00273] 165 [00274] 166 [00275] 167 [00276] 168 [00277] 169 [00278] 170 [00279] 171 [00280] 172 [00281] 173 [00282] 174 [00283] 175 [00284] 176 [00285] 177 [00286] 178 [00287] 179 [00288] 180 [00289] 181 [00290] 182 [00291] 183 [00292] 184 [00293] 185 [00294] 186 [00295] 187 [00296] 188 [00297] 189 [00298] 190 [00299]

[0233] In certain embodiments, the compound is a compound of Formula (I), (II), (III), (IV), (V), subgenera thereof, or species provided herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof. In certain embodiments, the compound is a compound of Formula (I), (II), (III), (IV), (V), subgenera thereof, or species provided herein, or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof. In certain embodiments, the compound is a compound of Formula (I), (II), (III), (IV), (V), subgenera thereof, or species provided herein, or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is a compound of Formula (I), (II), (III), (IV), (V), subgenera thereof, or species provided herein, or stereoisomer thereof. In certain embodiments, the compound is a compound of Formula (I), (II), (III), (IV), (V), subgenera thereof, or species provided herein, or tautomer thereof. In certain embodiments, the compound is the free base of compound of Formula (I), (II), (III), (IV), (V), subgenera thereof, or species provided herein.

Methods and Uses

[0234] As described herein, compounds of the disclosure (e.g., compounds of Formula (I)) exhibit activity as epithelial sodium channel (ENaC) blockers. Without being bound by any particular theory, it is believed that the compounds of the disclosure may function in vivo by blocking epithelial sodium channels present in mucosal surfaces and thereby reduce the absorption of sodium and water by the mucosal surfaces. This effect preserves the volume of protective liquids on mucosal surfaces and rebalances the system. As a consequence, the compounds of the disclosure are useful as medicaments, particularly for the treatment or prevention of clinical conditions for which a sodium channel blocker may be indicated. Such conditions include pulmonary conditions and conditions which are ameliorated by increased mucosal hydration in mucosal surfaces other than pulmonary mucosal surfaces. Further conditions include, but are not limited to, conditions of the skin. The present disclosure provides methods for treating each of these conditions in a subject (e.g., in a human) comprising administering to said subject a compound of the present disclosure, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition thereof.

[0235] The mucosal surfaces at the interface between the environment and the body have evolved a number of innate defense, i.e., protective mechanisms. A principal form of such innate defense is to cleanse these surfaces with liquid. Typically, the quantity of the liquid layer on a mucosal surface reflects the balance between epithelial liquid secretion, often reflecting anion (Cl.sup. and/or HCO.sub.3.sup.) secretion coupled with water (and a cation counter-ion), and epithelial liquid absorption, often reflecting Na.sup.+ absorption, coupled with water and counter anion (Cl.sup. and/or HCO.sub.3.sup.). Many diseases of mucosal surfaces are caused by too little protective liquid on those mucosal surfaces created by an imbalance between secretion (too little) and absorption (relatively too much). The defective salt transport processes that characterize these mucosal dysfunctions reside in the epithelial layer of the mucosal surface.

[0236] One approach to replenish the protective liquid layer on mucosal surfaces is to re-balance the system by blocking Na.sup.+ channel and liquid absorption. The epithelial protein that mediates the rate-limiting step of Na.sup.+ and liquid absorption is the epithelial Na.sup.+ channel (ENaC). ENaC is positioned on the apical surface of the epithelium, i.e., the mucosal surface-environmental interface. In some instances, to inhibit ENaC mediated Na.sup.+ and liquid absorption, an ENaC blocker can be delivered to the mucosal surface and maintained at this site to achieve maximum therapeutic benefit.

[0237] The use of ENaC blockers has been reported for a variety of diseases which are ameliorated by increased mucosal hydration. In particular, the use of ENaC blockers in the treatment of respiratory diseases such as for example chronic bronchitis (CB), cystic fibrosis (CF), and COPD, all of which reflect the body's failure to clear mucus normally from the lungs and ultimately result in chronic airway infection has been reported (R. C. Boucher, Journal of Internal Medicine 2007, 261(1), 5; R. C. Boucher, Trends in Molecular Medicine 2007 13(6), 231). ENaC blockers have also been reported as potential treatments for primary ciliary dyskinesia, non-cystic fibrosis bronchiectasis, and asthma. Further, mucus plugs and mucus dehydration are implicated in asthma. Mucus plugs may trap air in subjects who have this physiologic abnormality, thus are a plausible mechanism of chronic airflow obstruction in severe asthma, and EPO-generated oxidants may mediate mucus plug formation (Dunican, E. M. et al. Journal of Clinical Investigation 2018 128(3), 997).

[0238] Data indicate that the initiating problem in chronic bronchitis, primary ciliary dyskinesia, and cystic fibrosis is the failure to clear mucus from airway surfaces. The failure to clear mucus reflects an imbalance in the quantities of mucus as airway surface liquid (ASL) on airway surfaces. This imbalance results in a relative reduction in ASL which leads to mucus concentration, reduction in the lubricant activity of the periciliary liquid (PCL), mucus adherence to the airway surface, and failure to clear mucus via ciliary activity to the mouth. The reduction in mucus clearance leads to chronic bacterial colonization of mucus adherent to airway surfaces. The chronic retention of bacteria, inability of local antimicrobial substances to kill mucus-entrapped bacteria on a chronic basis, and the consequent chronic inflammatory response to this type of surface infection, are manifest in, e.g., chronic bronchitis, cystic fibrosis, and primary ciliary dyskinesia.

[0239] Too little protective surface liquid on other mucosal surfaces is a common pathophysiology of a number of diseases. For example, in xerostomia (dry mouth) the oral cavity is depleted of liquid due to a failure of the parotid sublingual and submandibular glands to secrete liquid despite continued Na.sup.+ (ENaC) transport mediated liquid absorption from the oral cavity. Keratoconjunctivitis sira (dry eye) is caused by failure of lacrimal glands to secrete liquid in the face of continued Na.sup.+ dependent liquid absorption on conjunctional surfaces. In rhinosinusitis, there is an imbalance between mucin secretion and relative ASL depletion. Failure to secrete Cl.sup. (and liquid) in the proximal small intestine, combined with increased Na.sup.+ (and liquid) absorption in the terminal ileum leads to the distal intestinal obstruction syndrome (DIOS). In older patients excessive Na.sup.+ (and volume) absorption in the descending colon produces constipation and diverticulitis.

[0240] Examples of ENaC inhibitors and methods of using the same can be found in, e.g., PCT Publication Nos. WO2003/070182, WO2003/070184, WO2004/073629, WO2005/025496, WO2005/016879, WO2005/018644, WO2006/022935, WO2006/023573, WO2006/023617, WO2007/018640, WO2007/146869, WO2008/031028, WO2008/031048, and U.S. Pat. Nos. 6,858,614, 6,858,615, 6,903,105, 7,064,129, 7,186,833, 7,189,719, 7,192,958, 7,192,959, 7,192,960, 7,241,766, 7,247,636, 7,247,637, 7,317,013, 7,332,496, 7,368,447, 7,368,450, 7,368,451, 7,375,102, 7,388,013, 7,399,766, 7,410,968, 7,807,834, 7,842,697, and 7,868,010.

[0241] Accordingly, in one aspect, provided herein are methods for blocking sodium channels in a subject comprising administering to the subject a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein.

[0242] In another aspect, provided herein are methods for promoting hydration of mucosal surfaces, improving mucociliary clearance, or restoring mucosal defense in a subject comprising administering to the subject a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein. In some embodiments, the method is for promoting hydration of mucosal surfaces in a subject. In some embodiments, the method is for improving mucociliary clearance. In some embodiments, the method is for restoring mucosal defense in a subject.

[0243] Also provided are methods for stimulating, enhancing, or improving mucociliary clearance in a subject comprising administering to a subject a compound as disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical compositions disclosed herein. Mucociliary clearance will be understood to include the natural mucociliary actions involved in the transfer or clearance of mucus in the airways, including the self-clearing mechanisms of the bronchi. Therefore, also provided is a method of improving mucus clearance in the airways of a subject.

[0244] In another aspect, provided herein are methods for treating and/or preventing a disease or disorder in a subject comprising administering to the subject a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein. In some embodiments, the method is for treating a disease or disorder in a subject. In certain embodiments, the method is for preventing a disease or disorder in a subject. In some embodiments, the method is for treating a disease or disorder selected from reversible or irreversible airway obstruction, chronic obstructive pulmonary disease (COPD), asthma, primary ciliary dyskinesia, bronchiectasis, bronchiectasis due to conditions other than cystic fibrosis, acute bronchitis, chronic bronchitis, post-viral cough, cystic fibrosis, idiopathic pulmonary fibrosis, pneumonia, panbronchiolitis, transplant-associate bronchiolitis, and ventilator-associated tracheobronchitis; or for preventing ventilator-associated pneumonia in a subject. In certain embodiments, the method is for treating a disease or disorder selected from dry mouth (xerostomia), dry skin, vaginal dryness, sinusitis, rhinosinusitis, nasal dehydration, nasal dehydration brought on by administering dry oxygen, dry eye, Sjogren's disease, otitis media, distal intestinal obstruction syndrome, esophagitis, constipation, mucus accumulation and inflammation, chronic diverticulitis, and fibrosis resulting from inflammation and/or oxidant stress in the airways driven by mucus accumulation. In some embodiments, the disease or disorder is reversible or irreversible airway obstruction. In certain embodiments, the disease or disorder is chronic obstructive pulmonary disease (COPD). In certain embodiments, the disease or disorder is asthma. In certain embodiments, the disease or disorder is bronchiectasis. In certain embodiments, the disease or disorder is bronchiectasis due to conditions other than cystic fibrosis. In certain embodiments, the disease or disorder is acute bronchitis. In certain embodiments, the disease or disorder is chronic bronchitis. In certain embodiments, the disease or disorder is post-viral cough. In some embodiments, the disease or disorder is idiopathic pulmonary fibrosis. In certain embodiments, the disease or disorder is cystic fibrosis. In certain embodiments. In certain embodiments, the disease or disorder is pneumonia. In certain embodiments, the disease or disorder is panbronchiolitis. In certain embodiments, the disease or disorder is transplant-associate bronchiolitis. In certain embodiments, the disease or disorder is ventilator-associated tracheobronchitis. In certain embodiments, the disease or disorder is dry mouth (xerostomia). In certain embodiments, the disease or disorder is dry skin. In certain embodiments, the disease or disorder is vaginal dryness. In certain embodiments, the disease or disorder is sinusitis. In certain embodiments, the disease or disorder is rhinosinusitis. In certain embodiments, the disease or disorder is nasal dehydration. In certain embodiments, the disease or disorder is nasal dehydration brought on by administering dry oxygen. In certain embodiments, the disease or disorder is dry eye. In certain embodiments, the disease or disorder is Sjogren's disease. In certain embodiments, the disease or disorder is otitis media. In certain embodiments, the disease or disorder is primary ciliary dyskinesia. In certain embodiments, the disease or disorder is distal intestinal obstruction syndrome. In certain embodiments, the disease or disorder is esophagitis. In certain embodiments, the disease or disorder is constipation. In some embodiments, the disease or disorder is mucus accumulation and inflammation. In certain embodiments, the disease or disorder is chronic diverticulitis. In some embodiments, the disease or disorder is fibrosis resulting from inflammation and/or oxidant stress in the airways driven by mucus accumulation.

[0245] In some embodiments, the method is for preventing ventilator-associated pneumonia in a subject. In some embodiments, the method is for preventing ventilator-associated pneumonia in a subject comprising administering to the subject a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein.

[0246] In some embodiments, the method is for preventing transplant associated bronchiolitis or ventilator associated tracheobronchitis.

[0247] Also provided herein are methods for promoting ocular or corneal hydration in a subject comprising administering to the subject a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein. In some embodiments, the method is for promoting ocular hydration in a subject. In some embodiments, the method is for promoting corneal hydration in a subject.

[0248] In another aspect, provided herein are methods for preventing, mitigating, and/or treating deterministic health effects to the respiratory tract and/or other bodily organs caused by respirable aerosols in a subject comprising administering to the subject a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein. In certain embodiments, the respirable aerosol is selected from a respirable aerosol containing radionuclides, dust, asbestos, and an infectious agent. Without wishing to be bound by a particular theory, in some embodiments, the compounds disclosed herein (e.g., a compound of Formula (I)) increase lung clearance and thus, inherently benefit lung health by accelerating the removal of noxious agents.

[0249] In another aspect, provided herein are methods for using a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein, as a potassium-sparing diuretic. In some embodiments, the method is a method for preventing low levels of potassium. In certain embodiments, the method is a method for treating heart failure. In some embodiments, the method is a method for treating ascites. In some embodiments, the method is a method for treating high blood pressure.

[0250] In another aspect, provided herein are methods for regulating blood pressure in a subject comprising administering to the subject a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein. In certain embodiments, the method is for increasing blood pressure in a subject. In certain embodiments, the method is for decreasing blood pressure in a subject.

[0251] In another aspect, provided herein are methods for using a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein, as an acid-sensing ion channel modulator. In some embodiments, the method is a method for treating a disorder or disease of the central nervous system. In some embodiments, the method is a method for treating a disorder or disease of the peripheral nervous system. In some embodiments, the method is a method for treating pain. In some embodiments, the method is a method for treating a neurological disease or disorder. In some embodiments, the method is a method for treating retinal disorders. In some embodiments, the method is a method for preventing seizures.

[0252] In another aspect, provided herein are methods for using a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein, as a viroporin modulator. In some embodiments, the method is a method for treating a viral infection. In some embodiments, the viral infection is caused by a coronavirus, poliovirus, influenza virus, human papillomavirus, human immunodeficiency virus, hepatitis virus, enterovirus, coxsackievirus, bovine ephemeral fever virus, chlorovirus, avian reovirus, or polyomavirus virus. In some embodiments, the viral infection is hepatitis C virus, HIV-1, human papillomavirus 16, influenza A virus, influenza B virus, influenza C virus, poliovirus, respiratory syncytial virus, or SARS-CoV (e.g., SARS-CoV-2) infection.

[0253] In another aspect, provided herein are methods for preventing, mitigating, and/or treating deterministic health effects to the respiratory tract and/or other bodily organs caused by respirable aerosols containing radionuclides in a subject comprising administering to the subject a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein. In certain embodiments, the method is for preventing deterministic health effects to the respiratory tract and/or other bodily organs caused by respirable aerosols containing radionuclides in a subject. In some embodiments, the method is for mitigating deterministic health effects to the respiratory tract and/or other bodily organs caused by respirable aerosols containing radionuclides in a subject. In certain embodiments, the method is for treating deterministic health effects to the respiratory tract and/or other bodily organs caused by respirable aerosols containing radionuclides in a subject.

[0254] In some embodiments, the radionuclides are selected from the group consisting of Cobalt-60, Cesium-137, Iridium-192, Radium-226, Phosphorus-32, Strontium-89 and 90, Iodine-125, Thallium-201, Lead-210, Thorium-234, Uranium-238, Plutonium, Cobalt-58, Chromium-51, Americium, and Curium. In certain embodiments, the radionuclides are from a radioactive disposal device. In some embodiments, the sodium channel blocker or pharmaceutically acceptable salt thereof is administered in an aerosol suspension of respirable particles which the individual inhales. In some embodiments, the sodium channel blocker or a pharmaceutically acceptable salt thereof is administered post-exposure to the radionuclides.

[0255] In certain embodiments, the respirable aerosols containing radionuclides are from nuclear attacks, such as detonation of radiological dispersal devices (RDD), or accidents, such as nuclear power plant disasters. A major concern associated with radioactive technologies is how to prevent, mitigate or treat potential deterministic health effects to the respiratory tract, primarily the lung. The greatest risk to the lungs following a radiological attack, such as a dirty bomb, results from the inhalation and retention of insoluble radioactive particles. As a result of radioactive particle retention, the cumulative exposure to the lung is significantly increased, ultimately resulting in pulmonary fibrosis/pneumonitis and potentially death. Insoluble particles cannot be systemically cleared by chelating agents because these particles are not in solution. The only method known to effectively reduce the radiation dose to the lungs from inhaled insoluble radioactive aerosols is bronchoalveolar lavage (BAL). However, BAL is a procedure that has many drawbacks. BAL is a highly invasive procedure that must be performed at specialized medical centers by trained pulmonologists. As such, a BAL procedure is expensive. Given the drawbacks of BAL, it is not a treatment option that would be readily and immediately available to persons in need of accelerated removal of radioactive particles, for example, in the event of a nuclear attack. In the event of a nuclear attack or a nuclear accident, immediate and relatively easily administered treatment for persons who have been exposed or who are at risk of being exposed is needed. Sodium channel blockers administered as an inhalation aerosol have been shown to restore hydration of airway surfaces. Such hydration of airway surfaces aids in clearing accumulated mucus secretions and associated particulate matter from the lung. As such, without being bound by any particular theory, it is believed that sodium channel blockers can be used to accelerate the removal of radioactive particles from airway passages.

[0256] In one embodiment the disclosure provides a method for the treatment of a condition which is ameliorated by increased mucosal hydration in a subject.

[0257] In one embodiment the disclosure provides a method for reducing the frequency, severity or duration of acute exacerbation of COPD or for the treatment of one or more symptoms of acute exacerbation of COPD in a subject.

[0258] In one embodiment, a compound disclosed herein (e.g., a compound of Formula (I)) is used to treat a pulmonary condition.

[0259] Also provided herein are compounds as disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for use in blocking sodium channels in a subject.

[0260] Also provided are compounds as disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for use in promoting hydration of mucosal surfaces, improving mucociliary clearance, or restoring mucosal defense in a subject.

[0261] Also provided are compounds as disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for use in stimulating, enhancing, or improving mucociliary clearance in subject.

[0262] Provided herein are compounds as disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for use as a medicament.

[0263] In certain embodiments, a compound as disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrugs thereof, and pharmaceutical compositions thereof, is for use in treating a disease or disorder in a subject. In certain embodiments, the disease or disorder is reversible or irreversible airway obstruction, chronic obstructive pulmonary disease (COPD), asthma, primary ciliary dyskinesia, bronchiectasis, acute bronchitis, chronic bronchitis, post-viral cough, idiopathic pulmonary fibrosis, cystic fibrosis, pneumonia, panbronchiolitis, transplant-associate bronchiolitis, or ventilator-associated tracheobronchitis; or for use in preventing ventilator-associated pneumonia, in a subject. In certain embodiments, the disease or disorder is dry mouth (xerostomia), dry skin, vaginal dryness, sinusitis, rhinosinusitis, nasal dehydration, dry eye, Sjogren's disease, otitis media, distal intestinal obstruction syndrome, esophagitis, constipation, mucus accumulation and inflammation, chronic diverticulitis, or fibrosis resulting from inflammation and/or oxidant stress in the airways driven by mucus accumulation. In certain embodiments, the compound or composition is for use in promoting ocular or corneal hydration in a subject. In some embodiments, the disease or disorder is reversible or irreversible airway obstruction. In certain embodiments, the disease or disorder is chronic obstructive pulmonary disease (COPD). In certain embodiments, the disease or disorder is asthma. In certain embodiments, the disease or disorder is bronchiectasis. In certain embodiments, the disease or disorder is bronchiectasis due to conditions other than cystic fibrosis. In certain embodiments, the disease or disorder is acute bronchitis. In certain embodiments, the disease or disorder is chronic bronchitis. In certain embodiments, the disease or disorder is post-viral cough. In some embodiments, the disease or disorder is idiopathic pulmonary fibrosis. In certain embodiments, the disease or disorder is cystic fibrosis. In certain embodiments, the disease or disorder is pneumonia. In certain embodiments, the disease or disorder is panbronchiolitis. In certain embodiments, the disease or disorder is transplant-associate bronchiolitis. In certain embodiments, the disease or disorder is ventilator-associated tracheobronchitis. In certain embodiments, the disease or disorder is dry mouth (xerostomia). In certain embodiments, the disease or disorder is dry skin. In certain embodiments, the disease or disorder is vaginal dryness. In certain embodiments, the disease or disorder is sinusitis. In certain embodiments, the disease or disorder is rhinosinusitis. In certain embodiments, the disease or disorder is nasal dehydration. In certain embodiments, the disease or disorder is nasal dehydration brought on by administering dry oxygen. In certain embodiments, the disease or disorder is dry eye. In certain embodiments, the disease or disorder is Sjogren's disease. In certain embodiments, the disease or disorder is otitis media. In certain embodiments, the disease or disorder is primary ciliary dyskinesia. In certain embodiments, the disease or disorder is distal intestinal obstruction syndrome. In certain embodiments, the disease or disorder is esophagitis. In certain embodiments, the disease or disorder is constipation. In some embodiments, the disease or disorder is mucus accumulation and inflammation. In certain embodiments, the disease or disorder is chronic diverticulitis. In some embodiments, the disease or disorder is fibrosis resulting from inflammation and oxidant/or stress in the airways driven by mucus accumulation.

[0264] Also provided are compounds as disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for use in preventing ventilator-associated pneumonia in a subject.

[0265] Also provided are compounds as disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for use in promoting ocular or corneal hydration in a subject

[0266] Also provided are compounds as disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for use in preventing, mitigating, and/or treating deterministic health effects to the respiratory tract and/or other bodily organs caused by respirable aerosols containing radionuclides in a subject.

[0267] Also provided herein are uses of the compounds disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for the manufacture of medicaments.

[0268] Further provided herein are uses of the compounds disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for the manufacture of a medicament for blocking sodium channels.

[0269] Also provided herein are uses of the compounds disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for the manufacture of a medicament for promoting hydration of mucosal surfaces, improving mucociliary clearance, or restoring mucosal defense.

[0270] Further provided herein are uses of the compounds disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for the manufacture of a medicament for treating or preventing a disease or disorder. In certain embodiments, the disease or disorder is reversible or irreversible airway obstruction, chronic obstructive pulmonary disease (COPD), asthma, primary ciliary dyskinesia, bronchiectasis, acute bronchitis, chronic bronchitis, post-viral cough, idiopathic pulmonary fibrosis, cystic fibrosis, pneumonia, panbronchiolitis, transplant-associate bronchiolitis, or ventilator-associated tracheobronchitis, or for preventing ventilator-associated pneumonia. In certain embodiments, the disease or disorder is dry mouth (xerostomia), dry skin, vaginal dryness, sinusitis, rhinosinusitis, nasal dehydration, dry eye, Sjogren's disease, otitis media, distal intestinal obstruction syndrome, esophagitis, constipation, mucus accumulation and inflammation, chronic diverticulitis, or fibrosis resulting from inflammation and/or oxidant stress in the airways driven by mucus accumulation. In certain embodiments, the medicament is for promoting ocular or corneal hydration. In some embodiments, the disease or disorder is reversible or irreversible airway obstruction. In certain embodiments, the disease or disorder is chronic obstructive pulmonary disease (COPD). In certain embodiments, the disease or disorder is asthma. In certain embodiments, the disease or disorder is bronchiectasis. In certain embodiments, the disease or disorder is bronchiectasis due to conditions other than cystic fibrosis. In certain embodiments, the disease or disorder is acute bronchitis. In certain embodiments, the disease or disorder is chronic bronchitis. In certain embodiments, the disease or disorder is post-viral cough. In certain embodiments, the disease or disorder is idiopathic pulmonary fibrosis. In certain embodiments, the disease or disorder is cystic fibrosis. In certain embodiments, the disease or disorder is pneumonia. In certain embodiments, the disease or disorder is panbronchiolitis. In certain embodiments, the disease or disorder is transplant-associate bronchiolitis. In certain embodiments, the disease or disorder is ventilator-associated tracheobronchitis. In certain embodiments, the disease or disorder is dry mouth (xerostomia). In certain embodiments, the disease or disorder is dry skin. In certain embodiments, the disease or disorder is vaginal dryness. In certain embodiments, the disease or disorder is sinusitis. In certain embodiments, the disease or disorder is rhinosinusitis. In certain embodiments, the disease or disorder is nasal dehydration. In certain embodiments, the disease or disorder is nasal dehydration brought on by administering dry oxygen. In certain embodiments, the disease or disorder is dry eye. In certain embodiments, the disease or disorder is Sjogren's disease. In certain embodiments, the disease or disorder is otitis media. In certain embodiments, the disease or disorder is primary ciliary dyskinesia. In certain embodiments, the disease or disorder is distal intestinal obstruction syndrome. In certain embodiments, the disease or disorder is esophagitis. In certain embodiments, the disease or disorder is constipation. In certain embodiments, the disease or disorder is mucus accumulation and inflammation. In certain embodiments, the disease or disorder is chronic diverticulitis. In some embodiments, the disease or disorder is fibrosis resulting from inflammation and/or oxidant stress in the airways driven by mucus accumulation.

[0271] Also provided are uses of the compounds as disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for the preparation of a medicament for preventing ventilator-associated pneumonia in a subject.

[0272] Also provided are uses of the compounds as disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for the preparation of a medicament for promoting ocular or corneal hydration in a subject.

[0273] Further provided herein are uses of the compounds disclosed herein (e.g., compounds of Formula (I)), and pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, and prodrugs thereof, and pharmaceutical compositions thereof, for the preparation of a medicament for preventing, mitigating, and/or treating deterministic health effects to the respiratory tract and/or other bodily organs caused by respirable aerosols containing radionuclides.

[0274] Also provided are methods, compounds, and uses for: (a) reducing exacerbations of COPD in a subject; (b) a method for reducing exacerbations of CF in a subject; (c) a method of improving lung function (FEV1) in a subject, (d) a method of improving lung function (FEV1) in a subject experiencing COPD, (e) a method of improving lung function (FEV1) in a subject experiencing CF, (f) a method of reducing airway infections in a subject. Also provided are methods, compounds, methods, and uses for: (a) reducing exacerbations in bronchiectasis other than cystic fibrosis, (b) reducing exacerbations of asthma, (c) reducing exacerbations of primary ciliary dyskinesia, (d) improving lung function in a patient experiencing asthma, primary ciliary dyskinesia, and/or bronchiectasis other than cystic fibrosis, and (e) a method of improving forced vital capacity (FVC) in a subject with idiopathic pulmonary fibrosis.

[0275] The compounds of the present disclosure may also be useful in methods for obtaining a sputum sample from a human. The method may be carried out by administering a compound disclosed herein to at least one lung of the patient, and then inducing and collecting a sputum sample from that human.

[0276] In other embodiments, the present disclosure provides each of the methods, compounds, and uses described herein with the additional benefit of minimizing or eliminating hyperkalemia in the recipient of the method. Also provided are embodiments comprising each of the methods described herein wherein an improved therapeutic index is achieved.

[0277] In certain embodiments, methods described herein are carried out by administering an effective amount of a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrugs thereof, or pharmaceutical compositions thereof, to a subject. In certain embodiments, the effective amount is a therapeutically effective amount. In certain embodiments, the effective amount is a prophylactically effective amount.

[0278] The compounds disclosed herein (e.g., compounds of Formula (I)), or pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, or prodrugs thereof, are also useful for treating airborne infections. Examples of airborne infections include, for example, RSV (Respiratory Syncytial Virus).

[0279] The compounds disclosed herein, or pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, or prodrugs thereof, are also useful for treating an anthrax infection.

[0280] The present disclosure also provides use of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, for prophylactic, post-exposure prophylactic, preventive, or therapeutic treatment against diseases or conditions caused by pathogens. In some embodiments, the pathogens may be a pathogen used in bioterrorism. Until convenient and effective treatments are available against every bioterrorism threat, there exists a strong need for preventative, prophylactic or therapeutic treatments which can prevent or reduce the risk of infection from pathogenic agents.

[0281] The present disclose also provides such methods of prophylactic treatment. In one aspect, a prophylactic treatment method is provided comprising administering a prophylactically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, to an individual in need of prophylactic treatment against infection from one or more airborne pathogens. A particular example of an airborne pathogen is anthrax.

[0282] In another aspect, a prophylactic treatment method is provided for reducing the risk of infection from an airborne pathogen which can cause a disease in a subject, said method comprising administering an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, to the lungs of the human who may be at risk of infection from the airborne pathogen but is asymptomatic for the disease, wherein the effective amount of a sodium channel blocker and optionally an osmolyte are sufficient to reduce the risk of infection in the human. A particular example of an airborne pathogen is anthrax.

[0283] In another aspect, a post-exposure prophylactic treatment or therapeutic treatment method is provided for treating infection from an airborne pathogen comprising administering an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, to the lungs of a subject in need of treatment against infection from an airborne pathogen. The pathogens which may be protected against by the prophylactic post exposure, rescue, and therapeutic treatment methods of the disclosure include any pathogens which may enter the body through the mouth, nose or nasal airways, thus proceeding into the lungs. Typically, the pathogens will be airborne pathogens, either naturally occurring or by aerosolization. The pathogens may be naturally occurring or may have been introduced into the environment intentionally after aerosolization or other method of introducing the pathogens into the environment. Many pathogens which are not naturally transmitted in the air have been or may be aerosolized for use in bioterrorism. The pathogens for which the compounds disclosed herein may be useful includes, but is not limited to, category A, B and C priority pathogens as set forth by the NIAID. These categories correspond generally to the lists compiled by the Centers for Disease Control and Prevention (CDC). As set up by the CDC, Category A agents are those that can be easily disseminated or transmitted person-to-person, cause high mortality, with potential for major public health impact. Category B agents are next in priority and include those that are moderately easy to disseminate and cause moderate morbidity and low mortality. Category C consists of emerging pathogens that could be engineered for mass dissemination in the future because of their availability, ease of production and dissemination and potential for high morbidity and mortality. Particular examples of these pathogens are anthrax and plague. Additional pathogens which may be protected against, or the infection risk therefrom reduced include influenza viruses, rhinoviruses, adenoviruses and respiratory syncytial viruses, and the like. A further pathogen which may be protected against is the coronavirus which is believed to cause severe acute respiratory syndrome (SARS).

[0284] In a further aspect, the present disclosure provides methods of treating a disease or disorder of the skin of a subject comprising administering to the subject a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein. In some embodiments, the disease or disorder of the skin is a skin wound (e.g., due to mechanical damage, chemical, or burns), skin lesion or ulcer (e.g., cold sores, shingles, acne), inflammatory diseases of the skin (e.g., lupus, psoriasis, eczema, rosacea), skin rash (e.g., contact dermatitis and diaper rash), or scarring. In some embodiments, the disease or disorder is selected from the group consisting of psoriasis, an inflammatory disease of the skin, a wound, a lesion of the skin, an ulcer of the skin, eczema, lupus, rosacea, a skin rash, a cold sore, shingles, and acne. In one embodiment, the disorder of the skin is psoriasis. In another embodiment of the present invention, the disorder of the skin is an inflammatory disease of the skin. In one embodiment, the disorder of the skin is a wound. In another embodiment of the present invention, the disorder of the skin is a lesion of the skin. In another embodiment of the present invention, the disorder of the skin is an ulcer of the skin. In yet another embodiment, the disorder of the skin is eczema. In one embodiment, the disorder of the skin is lupus. In another embodiment, the disorder of the skin is rosacea. In another embodiment of the present invention, the disorder of the skin is a skin rash. In another embodiment, the disorder of the skin is a cold sore. In another embodiment, the disorder of the skin is shingles. In another embodiment, the disorder of the skin is acne.

[0285] In a further aspect, the present disclosure provides a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein promote healing of epithelial surfaces, including skin.

[0286] In a further aspect, the present disclosure provides a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein for treating psoriasis and other dermatological diseases.

[0287] In a further aspect, the present disclosure provides a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein for treating dermal wounds caused by trauma, burns, or chemicals injury, or resulting from inflammatory diseases.

[0288] In some embodiments, the present disclosure provides a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, or pharmaceutical composition disclosed herein for use in the treatment of diseases associated with disorders of the skin in a subject in need thereof (e.g., in a mammal, e.g., in a human). In some embodiments, the compounds and compositions of the invention may be used in the manufacture of a medicament for the treatment of diseases associated with disorders of the skin. Examples of diseases and disorders of the skin are described herein.

[0289] In one embodiment, the compounds of the invention may be used in the manufacture of a medicament for the treatment of diseases associated with disorders of the skin.

[0290] In other embodiments, the present disclosure provides methods with the additional benefit of minimizing or eliminating hyperkalemia in the recipient of the method. Also provided are embodiments comprising each of the methods described herein wherein an improved therapeutic index is achieved.

Pharmaceutical Compositions, Kits, and Administration

[0291] The present disclosure provides pharmaceutical compositions comprising a compound provided herein (e.g., a compound of Formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof). The pharmaceutical composition may comprise one or more pharmaceutically acceptable carriers/excipients. In certain embodiments, a compound described herein is provided in an effective amount in the pharmaceutical composition. In certain embodiments, the effective amount is a therapeutically effective amount. In certain embodiments, the effective amount is a prophylactically effective amount.

[0292] Pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. In general, such preparatory methods include bringing the compound described herein (i.e., the active ingredient) into association with a carrier or excipient, and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping, and/or packaging the product into a desired single- or multi-dose unit.

[0293] Pharmaceutical compositions can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. A unit dose is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage, such as one-half or one-third of such a dosage.

[0294] Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition described herein will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered. The composition may comprise between 0.0001% and 100% (e.g., 0.1-100%, 0.001-1% (e.g., 0.0001%, 0.001%, 0.01%, 0.1%, 1%, 10%, 100%)) (w/w) active ingredient.

[0295] Pharmaceutically acceptable excipients used in the manufacture of provided pharmaceutical compositions include inert diluents, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and perfuming agents may also be present in the composition. In some embodiments, one of the excipients is a cyclodextrin.

[0296] Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, cornstarch, powdered sugar, lactose (e.g., anhydrous lactose, spray dried lactose), trehalose, leucine (L-leucine) and mixtures thereof.

[0297] Exemplary granulating and/or dispersing agents include potato starch, corn starch, tapioca starch, sodium starch glycolate, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose, and wood products, natural sponge, cation-exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof.

[0298] Exemplary surface active agents and/or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminum silicate)), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxy polymethylene, polyacrylic acid, acrylic acid polymer, and carboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g., carboxymethylcellulose sodium, powdered cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan (Tween 60), polyoxyethylene sorbitan monooleate (Tween 80), sorbitan monopalmitate (Span 40), sorbitan monostearate (Span 60), sorbitan tristearate (Span 65), glyceryl monooleate, sorbitan monooleate (Span 80), polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj 45), polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and Solutol), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., Cremophor), polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether (Brij 30)), poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, Pluronic F-68, poloxamer P-188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and/or mixtures thereof.

[0299] Exemplary binding agents include starch (e.g., cornstarch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husks, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate (Veegum), and larch arabogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohol, and/or mixtures thereof.

[0300] Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, antiprotozoan preservatives, alcohol preservatives, acidic preservatives, and other preservatives. In certain embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent.

[0301] Exemplary antioxidants include alpha tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.

[0302] Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, and the like), citric acid and salts and hydrates thereof (e.g., citric acid monohydrate), fumaric acid and salts and hydrates thereof, malic acid and salts and hydrates thereof, phosphoric acid and salts and hydrates thereof, and tartaric acid and salts and hydrates thereof. Exemplary antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.

[0303] Exemplary antifungal preservatives include butyl paraben, methyl paraben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid.

[0304] Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and phenylethyl alcohol.

[0305] Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.

[0306] Other preservatives include tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisol (BHA), butylated hydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus, Phenonip, methylparaben, Germall 115, Germaben II, Neolone, Kathon, and Euxyl.

[0307] Exemplary buffering agents include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and mixtures thereof.

[0308] Exemplary lubricating agents include magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behanate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, hydrophobic amino acids (e.g., L-leucine, L-isoleucine), leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.

[0309] Exemplary natural oils include almond, apricot kernel, avocado, babassu, bergamot, black current seed, borage, cade, camomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba, macademia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, sasquana, savoury, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, and wheat germ oils. Exemplary synthetic oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and mixtures thereof.

[0310] In some embodiments, the composition further comprises one or more agents selected from osmolytes, anti-inflammatory agents, anticholinergic agents, 0-agonists, CFTR modulators, P2Y2 receptor agonists, PY214 antagonist, peroxisome proliferator-activated receptor agonists, kinase inhibitors, mucoactive agents, hydrating agents, immune-modulatory agents, antiinfective agents, or antihistamines.

[0311] In one aspect, provided herein is a pharmaceutical composition comprising a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, and an osmolyte. In some embodiments, the osmolyte is hypertonic saline. In some embodiments, the osmolyte is a reduced sugar. In certain embodiments, the osmolyte is mannitol. In certain embodiments, the osmolyte is xylitol. In some embodiments, the osmolyte is an ionic sugar. In some embodiments, the ionic sugar is sodium gluconate. Further non-limiting examples of osmolytes are provided and discussed in more detail herein.

[0312] In some embodiments, the pharmaceutical composition further comprises an excipient. In some embodiments, the excipient is a cyclodextrin. Cyclodextrins are a family of molecules that comprise cyclic oligomers of glucose. In some embodiments, the cyclodextrin is selected from the group consisting of -cyclodextrin, -cyclodextrin, -cyclodextrin, a hydroxypropylated cyclodextrin (e.g., 2-hydroxypropyl--cyclodextrin, 2-hydroxypropyl--cyclodextrin), heptakis-2,6-di--methyl--cyclodextrin, heptakis-2,3,6-tris-O-methyl--cyclodextrin, and randomly methylated -cyclodextrin, crystalline methylated -cyclodextrin. In some embodiments, the pharmaceutical composition comprises a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof and a cyclodextrin. In some embodiments, the pharmaceutical composition comprises a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, hypertonic saline, and a cyclodextrin.

[0313] Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredients, the liquid dosage forms may comprise inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. In certain embodiments for parenteral administration, the conjugates described herein are mixed with solubilizing agents such as Cremophor, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof.

[0314] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, U.S.P., and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables. The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0315] In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form may be accomplished by dissolving or suspending the drug in an oil vehicle.

[0316] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, (c) humectants such as glycerol, (d) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarding agents such as paraffin, (f) absorption accelerators such as quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, (h) absorbents such as kaolin and bentonite clay, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may include a buffering agent.

[0317] Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art of pharmacology. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulating compositions which can be used include polymeric substances and waxes. Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[0318] The active ingredient can be in a micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings, and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active ingredient can be admixed with at least one inert diluent such as sucrose, lactose, or starch. Such dosage forms may comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may comprise buffering agents. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulating agents which can be used include polymeric substances and waxes.

[0319] Dosage forms for topical and/or transdermal administration of a compound described herein may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and/or patches. Generally, the active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier or excipient and/or any needed preservatives and/or buffers as can be required. Additionally, the present disclosure contemplates the use of transdermal patches, which often have the added advantage of providing controlled delivery of an active ingredient to the body. Such dosage forms can be prepared, for example, by dissolving and/or dispensing the active ingredient in the proper medium. Alternatively or additionally, the rate can be controlled by either providing a rate controlling membrane and/or by dispersing the active ingredient in a polymer matrix and/or gel.

[0320] Suitable devices for use in delivering intradermal pharmaceutical compositions described herein include short needle devices. Intradermal compositions can be administered by devices which limit the effective penetration length of a needle into the skin. Alternatively or additionally, conventional syringes can be used in the classical mantoux method of intradermal administration. Jet injection devices which deliver liquid formulations to the dermis via a liquid jet injector and/or via a needle which pierces the stratum corneum and produces a jet which reaches the dermis are suitable. Ballistic powder/particle delivery devices which use compressed gas to accelerate the compound in powder form through the outer layers of the skin to the dermis are suitable.

[0321] Formulations suitable for topical administration include, but are not limited to, liquid and/or semi-liquid preparations such as liniments, lotions, oil-in-water and/or water-in-oil emulsions such as creams, ointments, and/or pastes, and/or solutions and/or suspensions. Formulations for topical administration may further comprise one or more of the additional ingredients described herein.

[0322] Compositions for rectal or vaginal administration are typically suppositories which can be prepared by mixing the conjugates described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol, or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active ingredient.

[0323] A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation for buccal administration. Such formulations may, for example, be in the form of tablets and/or lozenges made using conventional methods. Alternately, formulations for buccal administration may comprise a powder and/or an aerosolized and/or atomized solution and/or suspension comprising the active ingredient. Such powdered, aerosolized, and/or atomized formulations, when dispersed, may have an average particle and/or droplet size in the range from about 0.01 to about 7 m (e.g., 0.01 to 4 m, 0.5 to 7 m, 0.01 to 1 m, 0.01 to 0.05 m, 1 to 5 m), and may further comprise one or more of the additional ingredients described herein.

[0324] A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation for ophthalmic administration. Such formulations may, for example, be in the form of eye drops including, for example, a solution and/or suspension of the active ingredient in an aqueous or oily liquid carrier or excipient. Such drops may further comprise buffering agents, salts, and/or one or more other of the additional ingredients described herein. Other ophthalmically-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form and/or in a liposomal preparation. Ear drops and/or eye drops are also contemplated as being within the scope of this disclosure.

[0325] A pharmaceutical composition described herein can be prepared, packaged, and/or sold in a formulation suitable for pulmonary administration via the buccal cavity (i.e., the mouth). Such a formulation may comprise dry particles which comprise the active ingredient and which have an average particle and/or droplet size (e.g., diameter) in the range from about 0.001 to about 7 m, about 0.01 to about 7 m, about 0.5 to about 7 m, or from about 1 to about 5 m. The most typical form of dry powder delivery is through a dry powder inhaler in which the patient's inhalation serves to transport and potentially disaggregate/deagglomerate the powder. Such compositions may be in the form of dry powders for administration using a device comprising a dry powder reservoir to which a stream of propellant can be directed to disperse the powder and/or using a self-propelling solvent/powder dispensing container such as a device comprising the active ingredient dissolve and/or suspended in a low-boiling propellant in a sealed container. Dry powder compositions may include a solid fine powder diluent such as sugar and are conveniently provided in a unit dose form. Compositions can be in the form of suspensions for administration using a device comprising a self-propelling solvent and/or low-boiling propellant. Low-boiling propellants generally include liquid propellants having a boiling point of below 65 F. at atmospheric pressure. The propellant may further comprise additional ingredients such as a liquid non-ionic and/or solid anionic surfactant and/or a solid diluent (which may have a particle size of the same order as particles comprising the active ingredient).

[0326] Pharmaceutical compositions described herein formulated for pulmonary delivery may provide the active ingredient in the form of droplets of a solution and/or suspension. Such formulations can be prepared, packaged, and/or sold as aqueous and/or dilute alcoholic solutions and/or suspensions, optionally sterile, comprising the active ingredient, and may conveniently be administered using any nebulization and/or atomization device. Such formulations may further comprise one or more additional ingredients including, but not limited to, a flavoring agent such as saccharin sodium, a volatile oil, a buffering agent, a surface active agent, and/or a preservative such as methylhydroxybenzoate. The droplets provided by this route of administration may have an average particle and/or droplet size (e.g., diameter) in the range from about 0.5 to about 7 m, preferably from about 1 to about 5 m.

[0327] Formulations described herein as being useful for pulmonary delivery are useful for intranasal delivery (i.e., delivery through the nose for pulmonary deposition) of a pharmaceutical composition described herein. An exemplary formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle and/or droplet size in the range from about 0.2 to about 500 m.

[0328] Formulations described herein may also be delivered to the nose. Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle and/or droplet size in the range from about 0.5 to about 7 m, preferably from about 1 to about 5 m. Such a formulation is administered by rapid inhalation through the nasal passage from a container of the powder held close to the nares.

[0329] In certain embodiments, a pharmaceutical composition disclosed herein is suitable for inhalation. In some embodiments, the inhalable pharmaceutical composition comprises a compound disclosed herein, hypertonic saline, and a cyclodextrin. In certain embodiments, a pharmaceutical composition disclosed herein is a solution for aerosolization and administration by nebulizer. In certain embodiments, a pharmaceutical composition disclosed herein is suitable for administration by metered dose inhaler. In certain embodiments, a pharmaceutical composition disclosed herein is suitable for administration by soft mist inhaler. In certain embodiments, a pharmaceutical composition disclosed herein is suitable for administration by dry powder inhaler. In certain embodiments, a pharmaceutical composition disclosed herein is a dry powder for administration by dry powder inhaler. In certain embodiments, a pharmaceutical composition disclosed herein is a solution for administration by soft mist inhaler. In certain embodiments, a pharmaceutical composition disclosed herein is a dry powder for administration by soft mist inhaler.

[0330] In some preferred embodiments, the composition is an inhalable pharmaceutical composition which is suitable for inhalation and delivery to the endobronchial space. Typically, such a composition is delivered using a nebulizer, pressurized metered dose inhaler (MDI), soft mist inhaler, or dry powder inhaler (DPI). The composition is typically in the form of an aerosol comprising particles for delivery. The aerosol formulation used in the methods of the present disclosure may be (i) a liquid (e.g., solution) suitable for administration by a nebulizer, soft mist inhaler, or MDI; (ii) a liquid suspension formulation for administration by a MDI, or (iii) a powder suitable for administration by a DPI. Additionally, the aerosol formulation used in the methods of the present disclosure may be a powder as a suspension or solid suitable for administration by a MDI (i.e., suspension) or DPI (i.e., suspension or solid). In some embodiments, the inhalable pharmaceutical composition comprises a compound disclosed herein, hypertonic saline, and a cyclodextrin.

[0331] Aerosols used to administer medicaments to the respiratory tract are typically polydispersethat is they are comprised of particles of many different sizes. The particle size distribution is typically described by the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation (GSD). For optimum drug delivery to the endobronchial space the MMAD is in the range from about 0.5 to about 10 m, from about 0.5 to about 7 m, and preferably from about 1 to about 5 m, and the GSD is less than 3, and preferably less than about 2. In some embodiments, the MMAD is about 0.5 to about 7 m. In preferred embodiments, the MMAD is from about 1 to about 5 m (e.g., about 3 m). In some embodiments, the GSD is equal to or less than about 2. In some preferred embodiments, the MMAD is about 3 m and the GSD is equal or less than 2. Aerosols having a MMAD above 10 m are generally too large when inhaled to have a significant portion of the mass reach the lungs. Aerosols with a GSD greater than about 3 are not preferred for lung delivery as they contain a higher percentage of the medicament outside of the respirable range. To achieve these particle sizes in powder formulation, the particles of the active ingredient may be size reduced using conventional techniques such as micronization or spray drying. Non-limiting examples of other processes or techniques that can be used to produce respirable particles include precipitation, supercritical fluid, and freeze drying. The desired fraction may be separated out by air classification or sieving. In one embodiment, the particles are crystalline. For liquid formulations, the particle size is determined by the selection of a particular model of nebulizer or inhaler (e.g., soft mist inhaler, dry powder inhaler, or MDI) along with characteristics of the liquid formulation.

[0332] Aerosol particle size distributions are determined using devices well known in the art. For example, a Next Generation Impactor (NGI), a multi-stage Anderson cascade impactor or other suitable method such as those specifically cited within the US Pharmacopeia Chapter 601 as characterizing devices for aerosols emitted from metered-dose and dry powder inhalers.

[0333] Dry powder compositions for delivery to the lung by inhalation may be formulated with excipient and/or carriers or they may be formulated without excipient and/or carrier and instead including only the active ingredients in a dry powder form having a suitable particle size for inhalation. Dry powder compositions may also contain a mix of the active ingredient and a suitable powder base (carrier/diluent/excipient substance) such as mono-, di- or poly-saccharides (e.g., lactose or starch). Lactose is typically a preferred excipient for dry powder formulations. When a solid excipient such as lactose is employed, generally the particle size of the excipient will be much greater than the active ingredient to aid the dispersion of the formulation in the inhaler.

[0334] Non-limiting examples of dry powder inhalers include reservoir multi-dose inhalers, pre-metered multi-dose inhalers, capsule-based inhalers, and single-dose disposable inhalers. A reservoir inhaler contains many doses (e.g. 60) in one container. Prior to inhalation, the patient actuates the inhaler which causes the inhaler to meter one dose of medicament from the reservoir and prepare it for inhalation. Examples of reservoir DPIs include but are not limited to the Turbohaler by AstraZeneca and the ClickHaler by Vectura.

[0335] In a pre-metered multi-dose inhaler, each individual dose has been manufactured in a separate container, and actuation of the inhaler prior to inhalation causes a new dose of drug to be released from its container and prepared for inhalation. Examples of multidose DPI inhalers include but are not limited to Diskus by GSK, Gyrohaler by Vectura, and Prohaler by Valois. During inhalation, the inspiratory flow of the patient accelerates the powder out of the device and into the oral cavity. For a capsule inhaler, the formulation is in a capsule and stored outside the inhaler. The patient puts a capsule in the inhaler, actuates the inhaler (punctures the capsule), then inhales. Examples include the Rotohaler (GlaxoSmithKline), Spinhaler (Novartis), HandiHaler (IB), and TurboSpin (PH&T). With single-dose disposable inhalers, the patient actuates the inhaler to prepare it for inhalation, inhales, then disposes of the inhaler and packaging. Examples include the Twincer (U Groningen), OneDose (GFE), and Manta Inhaler (Manta Devices).

[0336] Generally, dry powder inhalers utilize turbulent flow characteristics of the powder path to cause the excipient-drug aggregates to disperse, and the particles of active ingredient are deposited in the lungs. However, certain dry powder inhalers utilize a cyclone dispersion chamber to produce particles of the desired respirable size. In a cyclone dispersion chamber, the drug enters a coin shaped dispersion chamber tangentially so that the air path and drug move along the outer circular wall. As the drug formulation moves along this circular wall, it bounces around and agglomerates are broken apart by impact forces. The air path spirals towards the center of the chamber exiting vertically. Particles that have small enough aerodynamic sizes can follow the air path and exit the chamber. In effect, the dispersion chamber works like a small jet mill. Depending on the specifics of the formulation, large lactose particles may be added to the formulation to aid in the dispersion through impact with the API particles.

[0337] The Twincer single-dose disposable inhaler appears to operate using a coin-shaped cyclone dispersion chamber referred to as an air classifier. See, U.S. Published Patent Application No. 2006/0237010 to Rijksuniversiteit Groningen. Papers published by the University of Groningen, have stated that a 60 mg dose of pure micronized colistin sulfomethate could be effectively delivered as an inhalable dry powder utilizing this technology.

[0338] In preferred embodiments, the aerosol formulation is delivered as a dry powder using a dry powder inhaler wherein the particles emitted from the inhaler have an MMAD in the range of about 1 m to about 5 m and a GSD about less than 2.

[0339] Examples of suitable dry powder inhalers and dry powder dispersion devices for use in the delivery of compounds and compositions according to the present disclosure include but are not limited to those disclosed in U.S. Pat. Nos. 7,520,278, 7,322,354, 7,246,617, 7,231,920, 7,219,665, 7,207,330, 6,880,555, 5,522,385, 6,845,772, 6,637,431, 6,329,034, 5,458,135, 4,805,811, and U.S. Published Patent Application No. 2006/0237010.

[0340] In one embodiment, the pharmaceutical formulation according to the disclosure is a dry powder for inhalation which is formulated for delivery by a Diskus-type device. The Diskus device comprises an elongate strip formed from a base sheet having a plurality of recesses spaced along its length and a lid sheet hermetically, but peelably sealed thereto to define a plurality of containers, each container having therein an inhalable formulation containing a predetermined amount of active ingredient either alone or in admixture with one or more carriers or excipients (e.g., lactose) and/or other therapeutically active agents. In some embodiments, the strip is sufficiently flexible to be wound into a roll. In some embodiments, the lid sheet and base sheet have leading end portions which are not sealed to one another and at least one of the leading end portions is constructed to be attached to a winding means. Also, the hermetic seal between the base and lid sheets extends over their whole width. To prepare the dose for inhalation, the lid sheet may be peeled from the base sheet in a longitudinal direction from a first end of the base sheet.

[0341] In one embodiment, the pharmaceutical formulation according to the disclosure is a dry powder for inhalation which is formulated for delivery using a single-dose disposable inhaler, and particularly the Twincer inhaler. The Twincer inhaler comprises a foil laminate blister with one or more recesses and a lid sheet hermetically but peelably sealed thereto to define a plurality of containers. Each container has therein an inhalable formulation containing a predetermined amount of active ingredient(s) either alone or in admixture with one or more carriers or excipients (e.g., lactose). The lid sheet has a leading end portion which is constructed to project from the body of the inhaler. The patient operates the device and thereby administers the aerosol formulation by 1) removing the outer packaging overwrap, 2) pulling the foil tab to uncover the drug in the blister, and 3) inhaling the drug from the blister.

[0342] In another embodiment, the pharmaceutical formulation according to the disclosure is a dry powder for inhalation wherein the dry powder is formulated into microparticles as described in PCT Publication No. WO2009/015286 or WO2007/114881, both to NexBio. Such microparticles are generally formed by adding a counter ion to a solution containing a compound of the disclosure in a solvent, adding an antisolvent to the solution; and gradually cooling the solution to a temperature below about 25 C., to form a composition containing microparticles comprising the compound. The microparticles comprising the compound may then be separated from the solution by any suitable means such as sedimentation, filtration or lyophilization. Suitable counterions, solvents and antisolvents for preparing microparticles of the compounds of the invention are described in WO2009/015286.

[0343] In another embodiment, a pharmaceutical composition according to the disclosure is delivered as a dry powder (e.g., suspension) using a metered dose inhaler. Non-limiting examples of metered dose inhalers and devices include those disclosed in U.S. Pat. Nos. 5,261,538, 5,544,647, 5,622,163, 4,955,371, 3,565,070, 3,361,306 and 6,116,234 and 7,108,159. In a preferred embodiment, a compound of the disclosure, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is delivered as a suspension using a metered dose inhaler wherein the emitted particles have an MMAD that is in the range of about 1 m to about 5 m and a GSD that is less than about 2.

[0344] Liquid aerosol formulations for delivery to the endobronchial space or lung by inhalation may for example be formulated as aqueous solutions or suspensions for aerosols delivered from pressurized packs, such as metered dose inhalers, with the use of suitable liquefied propellants, soft mist inhalers, or nebulizers. Such aerosol compositions suitable for inhalation can be either a suspension or a solution and generally contain the active ingredient(s) together with a pharmaceutically acceptable carrier or diluent (e.g., water (distilled or sterile), saline, hypertonic saline, or ethanol) and optionally one or more other therapeutically active agents. In some embodiments, the pharmaceutical composition comprises a compound disclosed herein, hypertonic saline, and a cyclodextrin.

[0345] Aerosol compositions for delivery by pressurized metered dose inhalers typically further comprise a pharmaceutically acceptable propellant. Examples of such propellants include fluorocarbon or hydrogen-containing chlorofluorocarbon or mixtures thereof, particularly hydrofluoroalkanes. In some embodiments, the hydrofluoroalkane is dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, especially 1,1,1,2-tetrafluoroethane, especially 1,1,1,2,3,3,3,-heptafluoro-n-propane, or a mixture thereof. The aerosol composition may be excipient free or may optionally contain additional formulation excipients well known in the art such as surfactants (e.g., oleic acid or lecithin) and cosolvents (e.g., ethanol). Pressurized formulations will generally be retained in a canister (e.g., an aluminum canister) closed with a valve (e.g., a metering valve) and fitted into an actuator provided with a mouthpiece.

[0346] In another embodiment, a pharmaceutical composition according to the disclosure is delivered as a liquid using a metered dose inhaler. Non-limiting examples of metered dose inhalers and devices include those disclosed in U.S. Pat. Nos. 6,253,762, 6,413,497, 7,601,336, 7,481,995, 6,743,413, and 7,105,152. In a preferred embodiment, a compound of the disclosure, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is delivered as a dry powder using a metered dose inhaler wherein the emitted particles have an MMAD that is in the range of about 1 m to about 5 m and a GSD that is less than about 2.

[0347] In one embodiment the aerosol formulation is suitable for aerosolization by a jet nebulizer, ultrasonic nebulizer, or mesh nebulizers including static (passive) and vibrating (active) nebulizers. Liquid aerosol formulations for nebulization may be generated by solubilizing or reconstituting a solid particle formulation or may be formulated with an aqueous vehicle with the addition of agents such as acid or alkali, buffer salts, and isotonicity adjusting agents. They may be sterilized by in-process techniques such as filtration, or terminal processes such as heating in an autoclave or gamma irradiation. They may also be presented in non-sterile form.

[0348] Patients may be sensitive to the pH, osmolality, and ionic content of a nebulized solution. Therefore, these parameters should be adjusted to be compatible with the active ingredient and tolerable to patients. The most preferred solution or suspension of active ingredient will contain a chloride (e.g., NaCl) concentration>30 mM at pH 4.5-7.4, preferably 5.0-5.5, and an osmolality of from about 800-3200 mOsm/kg. The pH of the solution can be controlled by either titration with common acids (e.g., hydrochloric acid or sulfuric acid) or bases (e.g., sodium hydroxide) or via the use of buffers. Commonly used buffers include citrate buffers, such as citric acid/sodium citrate buffers, acetate buffers, such as acetic acid/sodium acetate buffers, and phosphate buffers.

[0349] Useful acetate, phosphate, and citrate buffers include sodium acetate, sodium acetate trihydrate, ammonium acetate, potassium acetate, sodium phosphate, sodium phosphate dibasic, disodium hydrogen phosphate, potassium dihydrogen phosphate, potassium hydrogen phosphate, potassium phosphate, sodium citrate, and potassium citrate. Other buffers which may be utilized include sodium hydroxide, potassium hydroxide, ammonium hydroxide, aminomethylpropanol, tromethamine, tetrahydroxypropyl ethylenediamine, citric acid, acetic acid, hydroxytricarboxylic acid or a salt thereof (e.g., a citrate or sodium citrate salt thereof), lactic acid, and salts of lactic acid (e.g., sodium lactate, potassium lactate, lithium lactate, calcium lactate, magnesium lactate, barium lactate, aluminum lactate, zinc lactate, silver lactate, copper lactate, iron lactate, manganese lactate, and ammonium lactate), monoethanolamine, diethanolamine, triethanolamine, diisopropanolamine, as well as combinations thereof, and the like.

[0350] Such formulations may be administered using commercially available nebulizers or other atomizers that can break the formulation into particles or droplets suitable for deposition in the respiratory tract. Non-limiting examples of nebulizers which may be employed for the aerosol delivery of a composition of the disclosure include jet nebulizers such as pneumatic jet nebulizers, vented or breath-enhanced jet nebulizers, and breath actuated jet nebulizers; ultrasonic nebulizers; or mesh nebulizers including static (passive) or active (vibrating) mesh nebulizers. Commercially available nebulizers include the Aeroneb Go nebulizer (Aerogen), LC PLUS (Pari Pharma), and eFlow nebulizer (Pari Pharma). In an ultrasonic nebulizer, vibration of a piezoelectric crystal creates surface instabilities in the drug reservoir that cause droplets to be formed. In porous plate nebulizers (mesh nebulizers), pressure fields generated by sonic energy force liquid through the mesh pores where it breaks into droplets by Rayleigh breakup. The sonic energy may be supplied by a vibrating horn or plate driven by a piezoelectric crystal, or by the mesh itself vibrating. Non-limiting examples of atomizers include any single or twin fluid atomizer or nozzle that produces droplets of an appropriate size. A single fluid atomizer works by forcing a liquid through one or more holes, where the jet of liquid breaks up into droplets. Twin fluid atomizers work by either forcing both a gas and liquid through one or more holes, or by impinging a jet of liquid against another jet of either liquid or gas.

[0351] A jet nebulizer utilizes a high velocity stream of air blasting up through a column of water to generate droplets. Particles unsuitable for inhalation impact on walls or aerodynamic baffles. A vented or breath enhanced nebulizer works in essentially the same way as a jet nebulizer except that inhaled air passes through the primary droplet generation area to increase the output rate of the nebulizer while the patient inhales.

[0352] The choice of nebulizer which aerosolizes the aerosol formulation is important in the administration of the active ingredient(s). Different nebulizers have differing efficiencies based their design and operation principle and are sensitive to the physical and chemical properties of the formulation. For example, two formulations with different surface tensions may have different particle size distributions. Additionally, formulation properties such as pH, osmolality, and permeant ion content can affect tolerability of the medication, so preferred embodiments conform to certain ranges of these properties.

[0353] In a preferred embodiment, the formulation for nebulization is delivered to the endobronchial space as an aerosol having an MMAD between about 1 m and about 5 m and a GSD less than 2 using an appropriate nebulizer. To be optimally effective and to avoid upper respiratory and systemic side effects, the aerosol should not have a MMAD greater than about 5 m and should not have a GSD greater than about 2. If an aerosol has an MMAD larger than about 5 m or a GSD greater than about 2, a large percentage of the dose may be deposited in the upper airways, thus decreasing the amount of drug delivered to the desired site in the lower respiratory tract. If the MMAD of the aerosol is smaller than about 1 m, then a large percentage of the particles may remain suspended in the inhaled air and may then be exhaled during expiration.

[0354] The compounds of the disclosure, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, may also be administered by transbronchoscopic lavage.

[0355] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with ordinary experimentation.

[0356] Compounds provided herein are typically formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compositions described herein will be decided by a physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject or organism will depend upon a variety of factors including the disease being treated and the severity of the disorder; the activity of the specific active ingredient employed; the specific composition employed; the age, body weight, general health, sex, and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific active ingredient employed; the duration of the treatment; drugs used in combination or coincidental with the specific active ingredient employed; and like factors well known in the medical arts.

[0357] The compounds and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), mucosal, nasal, bucal, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol. Specifically, contemplated routes are oral administration (e.g., oral inhalation of aerosol (i.e., for targeting pulmonary deposition)), intravenous administration (e.g., systemic intravenous injection), regional administration via blood and/or lymph supply, and/or direct administration to an affected site. In general, the most appropriate route of administration will depend upon a variety of factors including the nature of the agent (e.g., its stability in the environment of the gastrointestinal tract), and/or the condition of the subject (e.g., whether the subject is able to tolerate oral administration). In certain embodiments, the compound or pharmaceutical composition described herein is suitable for topical administration to the eye of a subject.

[0358] The exact amount of a compound required to achieve an effective amount may vary from subject to subject, depending, for example, on species, age, and general condition of a subject, severity of the side effects or disorder, identity of the particular compound, mode of administration, and the like. An effective amount may be included in a single dose (e.g., single oral dose) or multiple doses (e.g., multiple oral doses). In certain embodiments, when multiple doses are administered to a subject or applied to a tissue or cell, any two doses of the multiple doses include different or substantially the same amounts of a compound described herein. In certain embodiments, when multiple doses are administered to a subject or applied to a tissue or cell, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses a day, two doses a day, one dose a day, one dose every other day, one dose every third day, one dose every week, one dose every two weeks, one dose every three weeks, or one dose every four weeks. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is one dose per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is two doses per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses per day. In certain embodiments, when multiple doses are administered to a subject or applied to a tissue or cell, the duration between the first dose and last dose of the multiple doses is one day, two days, four days, one week, two weeks, three weeks, one month, two months, three months, four months, six months, nine months, one year, two years, three years, four years, five years, seven years, ten years, fifteen years, twenty years, or the lifetime of the subject, tissue, or cell. In certain embodiments, the duration between the first dose and last dose of the multiple doses is three months, six months, or one year. In certain embodiments, the duration between the first dose and last dose of the multiple doses is the lifetime of the subject, tissue, or cell. In certain embodiments, a dose (e.g., a single dose, or any dose of multiple doses) described herein includes independently between 0.1 g and 1 g, between 0.001 mg and 0.01 mg, between 0.01 mg and 0.1 mg, between 0.1 mg and 1 mg, between 0.3 mg and 1 mg, between 0.3 mg and 3 mg, between 0.1 mg and 3 mg, between 1 mg and 3 mg, between 3 mg and 10 mg, between 10 mg and 30 mg, between 30 mg and 100 mg, between 100 mg and 300 mg, between 100 mg and 200 mg, between 300 mg and 1,000 mg, or between 1 g and 10 g, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 1 mg and 3 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 3 mg and 10 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 10 mg and 30 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 30 mg and 100 mg, inclusive, of a compound described herein.

[0359] In certain embodiments, an effective amount of a compound for administration one or more times a day to a 70 kg adult human comprises about 0.0001 mg to about 3000 mg, about 0.0001 mg to about 2000 mg, about 0.0001 mg to about 100 mg, about 0.0001 mg to about 10 mg, about 0.001 mg to about 10 mg, about 0.01 mg to about 1 mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg, about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1 mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about 1000 mg, about 100 mg to about 1000 mg, about 100 mg to about 200 mg, or about 125 mg to about 175 mg of a compound per unit dosage form.

[0360] In certain embodiments, an effective amount of a compound for administration one or more times a day comprises from about 0.005 mg to about 25 mg, from about 0.05 mg to about 25 mg from about 0.075 mg to about 5 mg, from about 0.075 to about 0.1 mg, or about 0.085 mg of a compound per unit dosage form. In certain embodiments, an effective amount of a compound for administration one or more times a day comprises from about 0.0033 mg to about 16.67 mg, from about 0.033 mg to about 16.67 mg from about 0.05 mg to about 3.33 mg, from about 0.05 to about 0.1 mg, or about 0.057 mg of a compound per unit dosage form. In certain embodiments, an effective amount of a compound for administration one or more times a day comprises from about 0.0025 mg to about 12.5 mg, from about 0.025 mg to about 12.5 mg from about 0.0375 mg to about 2.5 mg, from about 0.035 to about 0.055 mg, or about 0.043 mg of a compound per unit dosage form.

[0361] In some embodiments, the compounds disclosed herein are administered to result in a daily dose from about 0.01 mg to about 50 mg. In some embodiments, the compounds disclosed herein are administered to result in a daily dose from about 0.1 mg to about 50 mg. In preferred embodiments, the compounds disclosed herein are administered to result in a daily dose from about 0.10 mg to about 10 mg, and more preferably from about 0.15 mg to about 10 mg. In some embodiments, the compounds disclosed herein result in a daily dose of about 0.10-0.25 mg.

[0362] In certain embodiments, the compounds of the invention may be administered orally or parenterally at dosage levels sufficient to deliver from about 0.001 mg/kg to about 100 mg/kg, from about 0.01 mg/kg to about 50 mg/kg, from about 0.1 mg/kg to about 40 mg/kg, from about 0.5 mg/kg to about 30 mg/kg, from about 0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect. In certain embodiments, the compounds of the invention may be administered orally or parenterally at dosage levels sufficient to deliver from about 0.14 g/kg to about 714 g/kg, from about 1.42 g/kg to about 714 g/kg, from about 1.42 g/kg to about 143 g/kg, from about 2.14 g/kg to about 143 g/kg, from about 2 g/kg to about 3 g/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.

[0363] A pharmaceutically effective dose administered topically to the airway surfaces of a subject (e.g., by inhalation) of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, for treatment of a 70 kg human may be in the range of from about 10 ng to about 250 mg. In another embodiment, the pharmaceutically effective dose may be from about 0.1 to about 1000 g. Typically, the daily dose administered topically to the airway surfaces will be an amount sufficient to achieve dissolved concentration of active agent on the airway surfaces of from about 10.sup.9, 10.sup.8, or 10.sup.7 to about 10.sup.4, 10.sup.3, 102, or 10.sup.1 Moles/liter, more preferably from about 10.sup.9 to about 10.sup.4 Moles/liter. The selection of the specific dose for a patient will be determined by the attendant physician, clinician or veterinarian of ordinary skill in the art based upon a number of factors including those noted above. In one particular embodiment the dose of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, for the treatment of a 70 kg human will be in the range of from about 10 nanograms (ng) to about 250 mg. In another embodiment, the effective dose would be from about 50 mg to about 250 mg. In another embodiment, the effective dose would be from about 100 mg to about 200 mg. In another embodiment, the effective dose would be from about 120 mg to about 180 mg. In another embodiment, the effective dose would be from about 125 mg to about 175 mg. In another embodiment, the effective dose would be from about 0.1 g to about 1,000 g. In one embodiment, the dose of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, for the treatment of a 70 kg human will be in the range of from about 0.5 g to about 0.5 mg. In a further embodiment the dose will be from about 0.5 g to about 60 g. In another embodiment, the pharmaceutically effective dose will be from about 1 to about 10 g. In another embodiment, the pharmaceutically effective dose will be from about 5 g to about 50 g. Another embodiment will have an effective dose of from about 10 g to about 40 g. In two further embodiments, the pharmaceutically effective dose will be from about 15 g to about 50 g from about 15 g to about 30 g, respectively. It will be understood that in each of these dose ranges, all incremental doses in the range are included. For instance, the 0.5-50 g range includes individual doses of: 0.5 g, 0.6 g, 0.7 g, 0.8 g, 0.9 g, 1.0 g, 1.1 g, 1.2 g, 1.3 g, 1.4 g, 1.5 g, 1.6 g, 1.7 g, 1.8 g, 1.9 g, 2.0 g, 2.1 g, 2.2 g, 2.3 g, 2.4 g, 2.5 g, 2.6 g, 2.7 g, 2.8 g, 2.9 g, 3.0 g, 3.1 g, 3.2 g, 3.3 g, 3.4 g, 3.5 g, 3.6 g, 3.7 g, 3.8 g, 3.9 g, 4.0 g, 4.1 g, 4.2 g, 4.3 g, 4.4 g, 4.5 g, 4.6 g, 4.7 g, 4.8 g, 4.9 g, 5.0 g, 5.1 g, 5.2 g, 5.3 g, 5.4 g, 5.5 g, 5.6 g, 5.7 g, 5.8 g, 5.9 g, 6.0 g, 6.1 g, 6.2 g, 6.3 g, 6.4 g, 6.5 g, 6.6 g, 6.7 g, 6.8 g, 6.9 g, 7.0 g, 7.1 g, 7.2 g, 7.3 g, 7.4 g, 7.5 g, 7.6 g, 7.7 g, 7.8 g, 7.9 g, 8.0 g, 8.1 g, 8.2 g, 8.3 g, 8.4 g, 8.5 g, 8.6 g, 8.7 g, 8.8 g, 8.9 g, 9.0 g, 9.1 g, 9.2 g, 9.3 g, 9.4 g, 9.5 g, 9.6 g, 9.7 g, 9.8 g, 9.9 g, 10.0 g, 10.1 g, 10.2 g, 10.3 g, 10.4 g, 10.5 g, 10.6 g, 10.7 g, 10.8 g, 10.9 g, 11.0 g, 11.1 g, 11.2 g, 11.3 g, 11.4 g, 11.5 g, 11.6 g, 11.7 g, 11.8 g, 11.9 g, 12.0 g, 12.1 g, 12.2 g, 12.3 g, 12.4 g, 12.5 g, 12.6 g, 12.7 g, 12.8 g, 12.9 g, 13.0 g, 13.1 g, 13.2 g, 13.3 g, 13.4 g, 13.5 g, 13.6 g, 13.7 g, 13.8 g, 13.9 g, 14.0 g, 14.1 g, 14.2 g, 14.3 g, 14.4 g, 14.5 g, 14.6 g, 14.7 g, 14.8 g, 14.9 g, 15.0 g, 15.1 g, 15.2 g, 15.3 g, 15.4 g, 15.5 g, 15.6 g, 15.7 g, 15.8 g, 15.9 g, 16.0 g, 16.1 g, 16.2 g, 16.3 g, 16.4 g, 16.5 g, 16.6 g, 16.7 g, 16.8 g, 16.9 g, 17.0 g, 17.1 g, 17.2 g, 17.3 g, 17.4 g, 17.5 g, 17.6 g, 17.7 g, 17.8 g, 17.9 g, 18.0 g, 18.1 g, 18.2 g, 18.3 g, 18.4 g, 18.5 g, 18.6 g, 18.7 g, 18.8 g, 18.9 g, 19.0 g, 19.1 g, 19.2 g, 19.3 g, 19.4 g, 19.5 g, 19.6 g, 19.7 g, 19.8 g, 19.9 g, 20.0 g, 20.1 g, 20.2 g, 20.3 g, 20.4 g, 20.5 g, 20.6 g, 20.7 g, 20.8 g, 20.9 g, 21.0 g, 21.1 g, 21.2 g, 21.3 g, 21.4 g, 21.5 g, 21.6 g, 21.7 g, 21.8 g, 21.9 g, 22.0 g, 22.1 g, 22.2 g, 22.3 g, 22.4 g, 22.5 g, 22.6 g, 22.7 g, 22.8 g, 22.9 g, 23.0 g, 23.1 g, 23.2 g, 23.3 g, 23.4 g, 23.5 g, 23.6 g, 23.7 g, 23.8 g, 23.9 g, 24.0 g, 24.1 g, 24.2 g, 24.3 g, 24.4 g, 24.5 g, 24.6 g, 24.7 g, 24.8 g, 24.9 g, 25.0 g, 25.1 g, 25.2 g, 25.3 g, 25.4 g, 25.5 g, 25.6 g, 25.7 g, 25.8 g, 25.9 g, 26.0 g, 26.1 g, 26.2 g, 26.3 g, 26.4 g, 26.5 g, 26.6 g, 26.7 g, 26.8 g, 26.9 g, 27.0 g, 27.1 g, 27.2 g, 27.3 g, 27.4 g, 27.5 g, 27.6 g, 27.7 g, 27.8 g, 27.9 g, 28.0 g, 28.1 g, 28.2 g, 28.3 g, 28.4 g, 28.5 g, 28.6 g, 28.7 g, 28.8 g, 28.9 g, 29.0 g, 29.1 g, 29.2 g, 29.3 g, 29.4 g, 29.5 g, 29.6 g, 29.7 g, 29.8 g, 29.9 g, 30.0 g, 30.1 g, 30.2 g, 30.3 g, 30.4 g, 30.5 g, 30.6 g, 30.7 g, 30.8 g, 30.9 g, 31.0 g, 31.1 g, 31.2 g, 31.3 g, 31.4 g, 31.5 g, 31.6 g, 31.7 g, 31.8 g, 31.9 g, 32.0 g, 32.1 g, 32.2 g, 32.3 g, 32.4 g, 32.5 g, 32.6 g, 32.7 g, 32.8 g, 32.9 g, 33.0 g, 33.1 g, 33.2 g, 33.3 g, 33.4 g, 33.5 g, 33.6 g, 33.7 g, 33.8 g, 33.9 g, 34.0 g, 34.1 g, 34.2 g, 34.3 g, 34.4 g, 34.5 g, 34.6 g, 34.7 g, 34.8 g, 34.9 g, 35.0 g, 35.1 g, 35.2 g, 35.3 g, 35.4 g, 35.5 g, 35.6 g, 35.7 g, 35.8 g, 35.9 g, 36.0 g, 36.1 g, 36.2 g, 36.3 g, 36.4 g, 36.5 g, 36.6 g, 36.7 g, 36.8 g, 36.9 g, 37.0 g, 37.1 g, 37.2 g, 37.3 g, 37.4 g, 37.5 g, 37.6 g, 37.7 g, 37.8 g, 37.9 g, 38.0 g, 38.1 g, 38.2 g, 38.3 g, 38.4 g, 38.5 g, 38.6 g, 38.7 g, 38.8 g, 38.9 g, 39.0 g, 39.1 g, 39.2 g, 39.3 g, 39.4 g, 39.5 g, 39.6 g, 39.7 g, 39.8 g, 39.9 g, 40.0 g, 40.1 g, 40.2 g, 40.3 g, 40.4 g, 40.5 g, 40.6 g, 40.7 g, 40.8 g, 40.9 g, 41.0 g, 41.1 g, 41.2 g, 41.3 g, 41.4 g, 41.5 g, 41.6 g, 41.7 g, 41.8 g, 41.9 g, 42.0 g, 42.1 g, 42.2 g, 42.3 g, 42.4 g, 42.5 g, 42.6 g, 42.7 g, 42.8 g, 42.9 g, 43.0 g, 43.1 g, 43.2 g, 43.3 g, 43.4 g, 43.5 g, 43.6 g, 43.7 g, 43.8 g, 43.9 g, 44.0 g, 44.1 g, 44.2 g, 44.3 g, 44.4 g, 44.5 g, 44.6 g, 44.7 g, 44.8 g, 44.9 g, 45.0 g, 45.1 g, 45.2 g, 45.3 g, 45.4 g, 45.5 g, 45.6 g, 45.7 g, 45.8 g, 45.9 g, 46.0 g, 46.1 g, 46.2 g, 46.3 g, 46.4 g, 46.5 g, 46.6 g, 46.7 g, 46.8 g, 46.9 g, 47.0 g, 47.1 g, 47.2 g, 47.3 g, 47.4 g, 47.5 g, 47.6 g, 47.7 g, 47.8 g, 47.9 g, 48.0 g, 48.1 g, 48.2 g, 48.3 g, 48.4 g, 48.5 g, 48.6 g, 48.7 g, 48.8 g, 48.9 g, 49.0 g, 49.1 g, 49.2 g, 49.3 g, 49.4 g, 49.5 g, 49.6 g, 49.7 g, 49.8 g, 49.9 g, 50 g. Additional exemplary individual doses include: 0.01 mg, 0.02 mg, 0.03 mg, 0.04 mg, 0.05 mg, 0.06 mg, 0.07 mg, 0.08 mg, 0.09 mg, 0.10 mg, 0.11 mg, 0.12 mg, 0.13 mg, 0.14 mg, 0.15 mg, 0.16 mg, 0.17 mg, 0.18 mg, 0.19 mg, 0.20 mg, 0.21 mg, 0.22 mg, 0.23 mg, 0.24 mg, 0.25 mg, 0.26 mg, 0.27 mg, 0.28 mg, 0.29 mg, 0.30 mg, 0.31 mg, 0.32 mg, 0.33 mg, 0.34 mg, 0.35 mg, 0.36 mg, 0.37 mg, 0.38 mg, 0.39 mg, 0.40 mg, 0.41 mg, 0.42 mg, 0.43 mg, 0.44 mg, 0.45 mg, 0.46 mg, 0.47 mg, 0.48 mg, 0.49 mg, 0.50 mg, 0.51 mg, 0.52 mg, 0.53 mg, 0.54 mg, 0.55 mg, 0.56 mg, 0.57 mg, 0.58 mg, 0.59 mg, 0.60 mg, 0.61 mg, 0.62 mg, 0.63 mg, 0.64 mg, 0.65 mg, 0.66 mg, 0.67 mg, 0.68 mg, 0.69 mg, 0.70 mg, 0.71 mg, 0.72 mg, 0.73 mg, 0.74 mg, 0.75 mg, 0.76 mg, 0.77 mg, 0.78 mg, 0.79 mg, 0.80 mg, 0.81 mg, 0.82 mg, 0.83 mg, 0.84 mg, 0.85 mg, 0.86 mg, 0.87 mg, 0.88 mg, 0.89 mg, 0.90 mg, 0.91 mg, 0.92 mg, 0.93 mg, 0.94 mg, 0.95 mg, 0.96 mg, 0.97 mg, 0.98 mg, 0.99 mg, 1.0 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2.0 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3.0 mg, 3.1 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.5 mg, 3.6 mg, 3.7 mg, 3.8 mg, 3.9 mg, 4.0 mg, 4.1 mg, 4.2 mg, 4.3 mg, 4.4 mg, 4.5 mg, 4.6 mg, 4.7 mg, 4.8 mg, 4.9 mg, 5.0 mg, 5.1 mg, 5.2 mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg, 5.8 mg, 5.9 mg, 6.0 mg, 6.1 mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7.0 mg, 7.1 mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg, 7.6 mg, 7.7 mg, 7.8 mg, 7.9 mg, 8.0 mg, 8.1 mg, 8.2 mg, 8.3 mg, 8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg, 8.8 mg, 8.9 mg, 9.0 mg, 9.1 mg, 9.2 mg, 9.3 mg, 9.4 mg, 9.5 mg, 9.6 mg, 9.7 mg, 9.8 mg, 9.9 mg, 10.0 mg, 10.1 mg, 10.2 mg, 10.3 mg, 10.4 mg, 10.5 mg, 10.6 mg, 10.7 mg, 10.8 mg, 10.9 mg, 11.0 mg, 11.1 mg, 11.2 mg, 11.3 mg, 11.4 mg, 11.5 mg, 11.6 mg, 11.7 mg, 11.8 mg, 11.9 mg, 12.0 mg, 12.1 mg, 12.2 mg, 12.3 mg, 12.4 mg, 12.5 mg, 12.6 mg, 12.7 mg, 12.8 mg, 12.9 mg, 13.0 mg, 13.1 mg, 13.2 mg, 13.3 mg, 13.4 mg, 13.5 mg, 13.6 mg, 13.7 mg, 13.8 mg, 13.9 mg, 14.0 mg, 14.1 mg, 14.2 mg, 14.3 mg, 14.4 mg, 14.5 mg, 14.6 mg, 14.7 mg, 14.8 mg, 14.9 mg, 15.0 mg, 15.1 mg, 15.2 mg, 15.3 mg, 15.4 mg, 15.5 mg, 15.6 mg, 15.7 mg, 15.8 mg, 15.9 mg, 16.0 mg, 16.1 mg, 16.2 mg, 16.3 mg, 16.4 mg, 16.5 mg, 16.6 mg, 16.7 mg, 16.8 mg, 16.9 mg, 17.0 mg, 17.1 mg, 17.2 mg, 17.3 mg, 17.4 mg, 17.5 mg, 17.6 mg, 17.7 mg, 17.8 mg, 17.9 mg, 18.0 mg, 18.1 mg, 18.2 mg, 18.3 mg, 18.4 mg, 18.5 mg, 18.6 mg, 18.7 mg, 18.8 mg, 18.9 mg, 19.0 mg, 19.1 mg, 19.2 mg, 19.3 mg, 19.4 mg, 19.5 mg, 19.6 mg, 19.7 mg, 19.8 mg, 19.9 mg, 20.0 mg, 20.1 mg, 20.2 mg, 20.3 mg, 20.4 mg, 20.5 mg, 20.6 mg, 20.7 mg, 20.8 mg, 20.9 mg, 21.0 mg, 21.1 mg, 21.2 mg, 21.3 mg, 21.4 mg, 21.5 mg, 21.6 mg, 21.7 mg, 21.8 mg, 21.9 mg, 22.0 mg, 22.1 mg, 22.2 mg, 22.3 mg, 22.4 mg, 22.5 mg, 22.6 mg, 22.7 mg, 22.8 mg, 22.9 mg, 23.0 mg, 23.1 mg, 23.2 mg, 23.3 mg, 23.4 mg, 23.5 mg, 23.6 mg, 23.7 mg, 23.8 mg, 23.9 mg, 24.0 mg, 24.1 mg, 24.2 mg, 24.3 mg, 24.4 mg, 24.5 mg, 24.6 mg, 24.7 mg, 24.8 mg, 24.9 mg, 25.0 mg, 25.1 mg, 25.2 mg, 25.3 mg, 25.4 mg, 25.5 mg, 25.6 mg, 25.7 mg, 25.8 mg, 25.9 mg, 26.0 mg, 26.1 mg, 26.2 mg, 26.3 mg, 26.4 mg, 26.5 mg, 26.6 mg, 26.7 mg, 26.8 mg, 26.9 mg, 27.0 mg, 27.1 mg, 27.2 mg, 27.3 mg, 27.4 mg, 27.5 mg, 27.6 mg, 27.7 mg, 27.8 mg, 27.9 mg, 28.0 mg, 28.1 mg, 28.2 mg, 28.3 mg, 28.4 mg, 28.5 mg, 28.6 mg, 28.7 mg, 28.8 mg, 28.9 mg, 29.0 mg, 29.1 mg, 29.2 mg, 29.3 mg, 29.4 mg, 29.5 mg, 29.6 mg, 29.7 mg, 29.8 mg, 29.9 mg, 30.0 mg, 30.1 mg, 30.2 mg, 30.3 mg, 30.4 mg, 30.5 mg, 30.6 mg, 30.7 mg, 30.8 mg, 30.9 mg, 31.0 mg, 31.1 mg, 31.2 mg, 31.3 mg, 31.4 mg, 31.5 mg, 31.6 mg, 31.7 mg, 31.8 mg, 31.9 mg, 32.0 mg, 32.1 mg, 32.2 mg, 32.3 mg, 32.4 mg, 32.5 mg, 32.6 mg, 32.7 mg, 32.8 mg, 32.9 mg, 33.0 mg, 33.1 mg, 33.2 mg, 33.3 mg, 33.4 mg, 33.5 mg, 33.6 mg, 33.7 mg, 33.8 mg, 33.9 mg, 34.0 mg, 34.1 mg, 34.2 mg, 34.3 mg, 34.4 mg, 34.5 mg, 34.6 mg, 34.7 mg, 34.8 mg, 34.9 mg, 35.0 mg, 35.1 mg, 35.2 mg, 35.3 mg, 35.4 mg, 35.5 mg, 35.6 mg, 35.7 mg, 35.8 mg, 35.9 mg, 36.0 mg, 36.1 mg, 36.2 mg, 36.3 mg, 36.4 mg, 36.5 mg, 36.6 mg, 36.7 mg, 36.8 mg, 36.9 mg, 37.0 mg, 37.1 mg, 37.2 mg, 37.3 mg, 37.4 mg, 37.5 mg, 37.6 mg, 37.7 mg, 37.8 mg, 37.9 mg, 38.0 mg, 38.1 mg, 38.2 mg, 38.3 mg, 38.4 mg, 38.5 mg, 38.6 mg, 38.7 mg, 38.8 mg, 38.9 mg, 39.0 mg, 39.1 mg, 39.2 mg, 39.3 mg, 39.4 mg, 39.5 mg, 39.6 mg, 39.7 mg, 39.8 mg, 39.9 mg, 40.0 mg, 40.1 mg, 40.2 mg, 40.3 mg, 40.4 mg, 40.5 mg, 40.6 mg, 40.7 mg, 40.8 mg, 40.9 mg, 41.0 mg, 41.1 mg, 41.2 mg, 41.3 mg, 41.4 mg, 41.5 mg, 41.6 mg, 41.7 mg, 41.8 mg, 41.9 mg, 42.0 mg, 42.1 mg, 42.2 mg, 42.3 mg, 42.4 mg, 42.5 mg, 42.6 mg, 42.7 mg, 42.8 mg, 42.9 mg, 43.0 mg, 43.1 mg, 43.2 mg, 43.3 mg, 43.4 mg, 43.5 mg, 43.6 mg, 43.7 mg, 43.8 mg, 43.9 mg, 44.0 mg, 44.1 mg, 44.2 mg, 44.3 mg, 44.4 mg, 44.5 mg, 44.6 mg, 44.7 mg, 44.8 mg, 44.9 mg, 45.0 mg, 45.1 mg, 45.2 mg, 45.3 mg, 45.4 mg, 45.5 mg, 45.6 mg, 45.7 mg, 45.8 mg, 45.9 mg, 46.0 mg, 46.1 mg, 46.2 mg, 46.3 mg, 46.4 mg, 46.5 mg, 46.6 mg, 46.7 mg, 46.8 mg, 46.9 mg, 47.0 mg, 47.1 mg, 47.2 mg, 47.3 mg, 47.4 mg, 47.5 mg, 47.6 mg, 47.7 mg, 47.8 mg, 47.9 mg, 48.0 mg, 48.1 mg, 48.2 mg, 48.3 mg, 48.4 mg, 48.5 mg, 48.6 mg, 48.7 mg, 48.8 mg, 38.9 mg, 49.0 mg, 49.1 mg, 49.2 mg, 49.3 mg, 49.4 mg, 49.5 mg, 49.6 mg, 49.7 mg, 49.8 mg, 39.9 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, or 250 mg. The foregoing suggested doses may be adjusted using conventional dose calculations if the compound is administered via a different route. Determination of an appropriate dose for administration by other routes is within the skill of those in the art in light of the foregoing description and the general knowledge in the art.

[0364] Delivery of an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, may entail delivery of a single dosage form or multiple unit doses which may be delivered contemporaneously or separate in time over a designated period, such as 24 hours. A dose of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, (alone or in the form of a composition comprising the same) may be administered from one to ten times per day. Typically, a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, (alone or in the form of a composition comprising the same) will be administered four, three, two, or once per day (24 hours).

[0365] In one preferred embodiment, the composition comprises a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, and one or more osmolytes. In another preferred embodiment, the composition comprises a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, and hypertonic saline. In another preferred embodiment, the composition comprises a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, and xylitol. In another preferred embodiment, the composition comprises a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, hypertonic saline, and xylitol. In some embodiments, the pharmaceutical composition is delivered as an aerosol formulation. In some embodiments, the pharmaceutical composition is delivered as a liquid aerosol formulation.

[0366] In one preferred embodiment, the composition comprises a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, one or more osmolytes, and an excipient. In another preferred embodiment, the composition comprises a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, hypertonic saline, and a cyclodextrin. In some embodiments, the pharmaceutical composition is delivered as an aerosol formulation. In some embodiments, the pharmaceutical composition is delivered as a liquid aerosol formulation.

[0367] Also encompassed by the disclosure are kits (e.g., pharmaceutical packs). The kits provided may comprise a pharmaceutical composition or compound described herein and a container (e.g., a vial, ampule, bottle, syringe, and/or dispenser package, or other suitable container). In some embodiments, provided kits may optionally further include a second container comprising a pharmaceutical excipient for dilution or suspension of a pharmaceutical composition or compound described herein. In some embodiments, the pharmaceutical composition or compound described herein provided in the first container and the second container are combined to form a single unit dosage form. Thus, in one aspect, provided are kits including a first container comprising a compound or pharmaceutical composition described herein. In certain embodiments, the kits are useful for treating a disease or condition in a subject in need thereof. In certain embodiments, the kits are useful for preventing a disease in a subject in need thereof. In some embodiments, the kits comprise a single unit dose or multiple unit doses. In certain embodiments, the kit may include a container system comprising one or more primary containers and one or more secondary containers.

[0368] In certain embodiments, a kit described herein further includes instructions for using the kit. A kit described herein may also include information as required by a regulatory agency such as the U.S. Food and Drug Administration (FDA). In certain embodiments, the information included in the kits is prescribing information. In certain embodiments, the kits provide instructions for treating a disease or condition in a subject in need thereof. In certain embodiments, the kits provide instructions for preventing a disease in a subject in need thereof. A kit described herein may include one or more additional pharmaceutical agents described herein as a separate composition.

[0369] Also provided is a kit comprising: i) a pharmaceutically effective amount of a compound disclosed herein, compound of the disclosure, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, ii) one or more pharmaceutically acceptable excipients, carriers, or diluents, and iii) instructions for administering the compound of group i) and the excipients, carriers, or diluents of group ii) to a subject. A subject includes any subject in need of the methods of treatment described herein (e.g., a subject in need thereof).

[0370] Further embodiments also comprise an aerosolization device selected from the group of a nebulizer (including (a) jet nebulizers (e.g., continuous nebulizer, breath enhanced nebulizer, or breath actuated nebulizer), (b) mesh nebulizers (e.g., passive (or static) nebulizer, active (or vibrating) nebulizer), (c) ultrasonic nebulizers)) and inhaler (including (a) a dry powder inhaler (e.g., active and passive dry powder inhalers, single unit-dose inhalers (e.g., Rotohaler, Handihaler), multiple unit-dose inhalers (e.g., Diskus), reservoir inhalers (e.g., Turbohaler)), (b) a metered dose inhaler (including pressurized, solution, and suspension metered dose inhalers), (c) soft mist inhalers. Other embodiments also comprise administration via other pulmonary drug delivery systems. In some embodiments, the excipient is a cyclodextrin. In some embodiments, the kit comprises a pharmaceutical composition comprising a compound disclosed herein, hypertonic saline, and a cyclodextrin.

[0371] In one embodiment a kit comprises: i) from about 10 ng to about 10 mg of a compound of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, per dose, ii) from about 1 to about 5 mL of diluent per dose, and iii) instructions for administering the compound of group i) and the diluent of group ii) to a subject. In a further embodiment, the diluent is from about 1 to about 5 mL of a saline solution, as described herein, per dose. In a further embodiment, the diluent is from about 1 to about 5 mL of a hypotonic saline solution per dose. In another embodiment, the diluent is from about 1 to about 5 mL of a hypertonic saline solution per dose. In a still further embodiment, the diluent is from about 1 to about 5 mL of sterile water per dose. In a still further embodiment, the diluent is from about 1 to about 5 mL of sterile water suitable for inhalation per dose.

[0372] Also provided is a kit comprising: i) a solution comprising a pharmaceutically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, dissolved in a pharmaceutically acceptable diluent, and ii) instructions for administering the solution of group i) to a subject.

[0373] Also provided is a kit comprising: i) a solution comprising from about 10 ng to about 10 mg of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, dissolved in a pharmaceutically acceptable diluent, and ii) instructions for administering the solution of group i) to a subject. In a further embodiment, the diluent is from about 1 to about 5 mL of a saline solution, as described herein, per dose.

[0374] Another embodiment comprises a kit comprising: i) a pharmaceutically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, in a dry powder formulation suitable for inhalation, ii) optionally, one or more pharmaceutically acceptable excipients or carriers suitable for inhalation, and iii) instructions for administering the compound of group i) and the excipients or carriers of group ii) to a subject. In some embodiments, the excipient is a cyclodextrin. In a further embodiment, the kit also comprises a dry powder inhaler suitable for delivering the dry powder formulation to a recipient. The dry powder inhaler may be, in additional embodiments, a single-dose inhaler or a multi-dose inhaler. Other embodiments comprise administration via other pulmonary drug delivery systems.

[0375] Also provided is a kit comprising: i) a solution comprising from about 10 ng to about 10 mg of a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, dissolved in a pharmaceutically acceptable diluent, ii) an excipient, and iii) instructions for administering the solution of group i) to a subject. In a further embodiment, the diluent is from about 1 to about 5 mL of a saline solution, as described herein, per dose. In some embodiments, the excipient is a cyclodextrin.

[0376] Further embodiments of each of the kits described herein includes those in which the amount of the compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is one of the effective dose ranges described herein, including: a) from about 0.1 g to about 1,000 g, b) from about 0.5 g to about 0.5 mg, and c) from about 0.5 g to about 50 g.

[0377] For each of the kits described above there is an additional embodiment in which the diluent is hypertonic saline of the concentrations described herein. In another embodiment for each kit the diluent is hypotonic saline of the concentrations described herein. In a further embodiment for each kit, the diluent is sterile water suitable for inhalation.

Combinations

[0378] A compound or composition, as described herein, can be administered in combination with one or more additional pharmaceutical agents (e.g., therapeutically and/or prophylactically active agents). The compounds or compositions can be administered in combination with additional pharmaceutical agents that improve their activity (e.g., activity (e.g., potency and/or efficacy) in treating a disease in a subject in need thereof, in preventing a disease in a subject in need thereof, in reducing the risk to develop a disease in a subject in need thereof), improve bioavailability, improve safety, reduce drug resistance, reduce and/or modify metabolism, inhibit excretion, and/or modify distribution in a subject or cell. It will also be appreciated that the therapy employed may achieve a desired effect for the same disorder, and/or it may achieve different effects. In certain embodiments, the combination of a compound or composition described herein with an additional pharmaceutical agent shows a synergistic effect that is absent when either are used without the other. In some embodiments, the additional pharmaceutical agent achieves a desired effect for the same disorder. In some embodiments, the additional pharmaceutical agent achieves different effects.

[0379] In some embodiments, the additional pharmaceutical agent is delivered in a kit. In some embodiments, the kit further comprises a compound disclosed herein (e.g., a compound of Formula (I)), or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrugs thereof, or composition thereof.

[0380] The compound or composition can be administered concurrently with, prior to, or subsequent to one or more additional pharmaceutical agents, which may be useful as, e.g., combination therapies. The one or more additional pharmaceutical agents can be administered in the same composition or in different compositions. Pharmaceutical agents include therapeutically active agents. Pharmaceutical agents also include prophylactically active agents. Pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved for human or veterinary use by the U.S. Food and Drug Administration as provided in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells.

[0381] The additional pharmaceutical agents include, but are not limited to, anti-proliferative agents, anti-cancer agents, anti-angiogenesis agents, steroidal or non-steroidal anti-inflammatory agents (NSAIDs), immunosuppressants, anti-bacterial agents, anti-viral agents, cardiovascular agents, cholesterol-lowering agents, anti-diabetic agents, anti-allergic agents, contraceptive agents, pain-relieving agents, anesthetics, anti-coagulants, inhibitors of an enzyme, steroidal agents, steroidal or antihistamine, antigens, vaccines, antibodies, decongestant, sedatives, opioids, analgesics, anti-pyretics, and hormones.

[0382] Non-limiting examples of therapeutically active agents which may be formulated or used in combination with the compounds of the disclosure include but are not limited to osmolytes, anti-inflammatory agents, anticholinergic agents, -agonists (including selective .sub.2-agonists), P2Y2 receptor agonists, P2Y14 antagonists, peroxisome proliferator-activated receptor (PPAR) agonists, kinase inhibitors, mucoactive agents, hydrating agents, immune-modulatory agents, antiinfective agents, and antihistamines.

[0383] Use of the compounds of the disclosure (e.g., compounds of Formula (I)), or pharmaceutically acceptable salts, stereoisomers, tautomers, isotopically labeled derivatives, solvates, hydrates, polymorphs, co-crystals, or prodrug thereof, in combination with one or more other therapeutically active agents may lower the dose of the compound that is required to sufficiently hydrate mucosal surfaces, thereby reducing the potential for undesired side-effects attributable to systemic blocking of sodium channels such as for example in the kidneys.

[0384] In certain embodiments, a compound or compositions described herein is used in combination with an osmolyte. Osmolytes according to the present disclosure are molecules or compounds that are osmotically active. Osmotically active molecules and compounds are membrane-impermeable (i.e., essentially non-absorbable) on the airway or pulmonary epithelial surface. The terms airway surface and pulmonary surface, as used herein, include pulmonary airway surfaces such as the bronchi and bronchioles, alveolar surfaces, and nasal and sinus surfaces. Suitable osmolytes include ionic osmolytes (i.e., salts (e.g., ionic sugars (e.g. sodium gluconate)), and non-ionic osmolytes (i.e., sugars (e.g., fructose, galactose, glucose, dextrose, lactose, maltose, xylose, sucrose), a reduced sugar (e.g., glycerol, erythritol, threitol, D-threitol, L-threitol, xylitol, ribitol, arabitol, D-arabitol, L-arabitol, D-xylitol, mannitol, sorbitol, galactitol, allitol, altritol, L-sorbitol, L-mannitol), sugar alcohols (e.g., mannitol, xylitol, sorbitol, lactitol, erythritol, glycerol, threitol, arabitol, ribitol, galactitol, fucitol, maltitol, isomalt) and organic osmolytes). Osmolytes suitable for use in the present disclosure may be in racemic form or in the form of an enantiomer, diastereomer, tautomer, polymorph, or pseudopolymorph.

[0385] Examples of ionic osmolytes useful in the present disclosure include any salt of a pharmaceutically acceptable anion and a pharmaceutically acceptable cation. Preferably, either (or both) of the anion and cation are osmotically active and not subject to rapid active transport, in relation to the airway surfaces to which they are administered. Such compounds include but are not limited to anions and cations that are contained in FDA approved commercially marketed salts, see, e.g., Remington: The Science and Practice of Pharmacy, Vol. II, pg. 1457 (19.sup.th Ed. 1995), and can be used in any combination as known in the art.

[0386] Specific examples of pharmaceutically acceptable osmotically active anions include but are not limited to: acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsylate (camphorsulfonate), carbonate, chloride, citrate, dihydrochloride, edetate, edisylate (1,2-ethanedisulfonate), estolate (lauryl sulfate), esylate (1,2-ethanedisulfonate), fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate (p-glycollamidophenylarsonate), hexylresorcinate, hydrabamine (N,N-Di(dehydroabietyl) ethylenediamine), hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, nitrite, pamoate (embonate), pantothenate, phosphate or diphosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, teoclate (8-chlorotheophyllinate), triethiodide, and bicarbonate. Preferred anions include chloride, sulfate, nitrate, gluconate, iodide, bicarbonate, bromide, and phosphate.

[0387] Specific examples of pharmaceutically acceptable osmotically active cations include but are not limited to: organic cations (e.g., benzathine (N,N-dibenzylethylenediamine), chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methyl D-glucamine), procaine, D-lysine, L-lysine, D-arginine, L-arginine, triethylammonium, N-methyl D-glycerol, and the like) and metallic cations (e.g., aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, iron, ammonium, and the like). Preferred organic cations include 3-carbon, 4-carbon, 5-carbon and 6-carbon organic cations. Preferred cations include sodium, potassium, choline, lithium, meglumine, D-lysine, ammonium, magnesium, and calcium.

[0388] Specific examples of ionic osmolytes that may be used in combination with a compound of the disclosure include but are not limited to, sodium chloride (particularly hypertonic saline), potassium chloride, choline chloride, choline iodide, lithium chloride, meglumine chloride, L-lysine chloride, D-lysine chloride, ammonium chloride, potassium sulfate, potassium nitrate, potassium gluconate, potassium iodide, ferric chloride, ferrous chloride, potassium bromide, and combinations thereof. In one embodiment, provided herein is a combination of a compound of the disclosure, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, and an osmotically active salt. In one embodiment, provided herein is a combination of a compound of the disclosure, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, and two different osmotically active salts. When different salts are used, the anion or cation may be the same among the differing salts. Hypertonic saline is a preferred ionic osmolyte for use in combination with the compounds disclosed herein.

[0389] Non-ionic osmolytes include sugars, sugar-alcohols, and organic osmolytes. Sugars and sugar-alcohols useful as osmolytes in the present disclosure include but are not limited to: 3-carbon sugars (e.g., glycerol, dihydroxyacetone), 4-carbon sugars (e.g., both the D and L forms of erythrose, threose, and erythrulose), 5-carbon sugars (e.g., both the D and L forms of ribose, arabinose, xylose, lyxose, psicose, fructose, sorbose, and tagatose), 6-carbon sugars (e.g., both the D and L forms of altose, allose, glucose, mannose, gulose, idose, galactose, and talose, and the D and L forms of allo-heptulose, allo-hepulose, gluco-heptulose, manno-heptulose, gulo-heptulose, ido-heptulose, galacto-heptulose, talo-heptulose), and sugar-alcohols thereof. Additional sugars useful in the practice of the present disclosure include raffinose, raffinose series oligosaccharides, and stachyose. Both the D and L forms of the reduced form of each sugar/sugar alcohol are also suitable for the present disclosure. For example, glucose, when reduced, becomes sorbitol; an osmolyte within the scope of the disclosure. Accordingly, sorbitol and other reduced forms of sugar/sugar alcohols (e.g., mannitol, dulcitol, arabitol) are suitable osmolytes for use in the present disclosure. In some embodiments, mannitol is a preferred non-ionic osmolyte for use in combination with the compounds disclosed herein. In some embodiments, xylitol is a preferred non-ionic osmolyte for use in combination with the compounds disclosed herein.

[0390] Organic osmolytes generally refers to molecules that control intracellular osmolality in the kidney. See e.g., J. S. Handler et al., Comp. Biochem. Physiol, 117, 301-306 (1997); M. Burg, Am. J. Physiol. 268, F983-F996 (1995). Organic osmolytes include but are not limited to three major classes of compounds: polyols (polyhydric alcohols), methylamines, and amino acids. Suitable polyol organic osmolytes include but are not limited to inositol, myo-inositol, and sorbitol. Suitable methylamine organic osmolytes include but are not limited to choline, betaine, carnitine (L-, D- and DL forms), phosphorylcholine, lyso-phosphorylcholine, glycerophosphorylcholine, creatine, and creatine phosphate. Suitable amino acid organic osmolytes include but are not limited to the D- and L-forms of glycine, alanine, glutamine, glutamate, aspartate, proline, and taurine. Additional organic osmolytes suitable for use in the present disclosure include trehalose and sarcosine. Mammalian organic osmolytes are preferred, with human organic osmolytes being most preferred. However, certain organic osmolytes are of bacterial, yeast, and marine animal origin, and these compounds may also be employed.

[0391] Osmolyte precursors may be used in combination with the compounds of the disclosure. An osmolyte precursor as used herein refers to a compound which is converted into an osmolyte by a metabolic step, either catabolic or anabolic. Examples of osmolyte precursors include but are not limited to glucose, glucose polymers, glycerol, choline, phosphatidylcholine, lyso-phosphatidylcholine and inorganic phosphates, which are precursors of polyols and methylamines. Precursors of amino acid osmolytes include proteins, peptides, and polyamino acids, which are hydrolyzed to yield osmolyte amino acids, and metabolic precursors which can be converted into osmolyte amino acids by a metabolic step such as transamination. For example, a precursor of the amino acid glutamine is poly-L-glutamine, and a precursor of glutamate is poly-L-glutamic acid.

[0392] Chemically modified osmolytes or osmolyte precursors may also be employed in the formulations, uses, regimens, and kits described herein. Such chemical modifications involve linking the osmolyte, or precursor thereof, to an additional chemical group which alters or enhances the effect of the osmolyte or osmolyte precursor (e.g., inhibits degradation of the osmolyte molecule). Such chemical modifications have been utilized with drugs or prodrugs and are known in the art. (See, for example, U.S. Pat. Nos. 4,479,932 and 4,540,564; Shek, E. et al., J. Med. Chem. 19:113-117 (1976); Bodor, N. et al., J. Pharm. Sci. 67:1045-1050 (1978); Bodor, N. et al., J. Med. Chem. 26:313-318 (1983); Bodor, N. et al., J. Pharm. Sci. 75:29-35 (1986); each of which are incorporated herein by reference).

[0393] Preferred osmolytes for use in combination with the compounds of the disclosure include sodium chloride, particularly hypertonic saline, mannitol, xylitol, and sodium gluconate.

[0394] In some embodiments, 7% or >7% hypertonic saline is used in the formulation. For the formulation of 7% and >7% hypertonic saline, formulations containing bicarbonate anions may be particularly useful, especially for respiratory disorders with cystic fibrosis transmembrane conductance regulator (CFTR) dysfunction such as CF or COPD. Recent findings indicate that, although the relative ratio of HCO.sub.3 conductance/Cl.sup. conductance is between 0.1 and 0.2 for single CFTR channels activated with cAMP and ATP, the ratio in the sweat duct can range from virtually 0 to almost 1.0, depending on conditions of stimulation. That is, combining cAMP+cGMP+-ketoglutarate can yield CFTR HCO.sub.3 conductance almost equal to that of Cl.sup. conductance (Quiton et al. Physiology, Vol. 22, No. 3, 212-225, June 2007). Furthermore, formulations of 7% and >7% hypertonic saline containing bicarbonate anions may be particularly useful due to better control of the pH in the airway surface liquid. First, it has shown that that airway acidification occurs in CF (Tate, S. et al. Thorax, 2002. 57(11), 926) and that absent CFTR-dependent bicarbonate secretion can lead to an impaired capacity to respond to airway conditions associated with acidification of airway surface liquid layer (Coakley, R. D. et al Proc. Natl. Acad. Sci. USA, 2003. 100(26), 16083). Second, addition of hypertonic saline solution without bicarbonate to the surface of the lung may further dilute the bicarbonate concentrations, and potentially reduce the pH or the ability to respond to airway acidification within the airway surface liquid layer. Therefore, addition of bicarbonate anions to hypertonic saline may help maintain or improve the pH of airway surface liquid layer in CF patients. Due to this evidence, inclusion of bicarbonate anion in the formulation of 7% or >7% hypertonic saline administered by a method disclosed herein would be particularly useful. Formulations comprising up to 30 to 200 mM concentrations of bicarbonate anions are of particular interest for formulations comprising 7% or >7% hypertonic saline solutions and a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof.

[0395] Hypertonic saline is understood to have a salt concentration greater than that of normal saline (NS) (i.e. greater than 9 g/L or 0.9% w/v) and hypotonic saline has a salt concentration less than that of normal saline, such as from about 1 g/L or 0.1% w/v to about 8 g/L or 0.8% w/v. Hypertonic saline solutions useful in the formulations and methods of treatment herein may have a salt concentration from about 1% to about 23.4% (w/v). In one embodiment, the hypertonic saline solution has a salt concentration from about 60 g/L (6% w/v) to about 100 g/L (10% w/v). In another embodiment, the saline solution has a salt concentration from about 70 g/L (7% w/v) to about 100 g/L (10% w/v). In further embodiments, the saline solution has a salt concentration of: a) from about 0.5 g/L (0.05% w/v) to about 70 g/L (7% w/v), b) from about 1 g/L (0.1% w/v) to about 60 g/L (6% w/v), c) from about 1 g/L (0.1% w/v) to about 50 g/L (5% w/v), d) from about 1 g/L (0.1% w/v) to about 40 g/L (4% w/v), e) from about 1 g/L (0.1% w/v) to about 30 g/L (3% w/v), or f) from about 1 g/L (0.1% w/v) to about 20 g/L (2% w/v).

[0396] Specific concentrations of saline solutions useful in the formulations, uses, regimens, and kits described herein include, independently, those having salt concentrations of 1 g/L (0.1% w/v), 2 g/L (0.2% w/v), 3 g/L (0.3% w/v), 4 g/L (0.4% w/v), 5 g/L (0.5% w/v), 6 g/L (0.6% w/v), 7 g/L (0.7% w/v), 8 g/L (0.8% w/v), 9 g/L (0.9% w/v), 10 g/L (1% w/v), 20 g/L (2% w/v), 30 g/L (3% w/v), 40 g/L (4% w/v), 50 g/L (5% w/v), 60 g/L (6% w/v), 70 g/L (7% w/v), 80 g/L (8% w/v), 90 g/L (9% w/v), 100 g/L (10% w/v), 110 g/L (11% w/v), 120 g/L (12% w/v), 130 g/L (13% w/v), 140 g/L (14% w/v), 150 g/L (15% w/v), 160 g/L (16% w/v), 170 g/L (17% w/v), 180 g/L (18% w/v), 190 g/L (19% w/v), 200 g/L (20% w/v), 210 g/L (21% w/v), 220 g/L (22% w/v), and 230 g/L (23% w/v). Saline concentrations between each of these listed concentrations/percentages may also be used, such as saline of 1.7 g/L (0.17% w/v), 1.25 g/L (1.25% w/v), 1.5 g/L (1.5% w/v), 25 g/L (2.5% w/v), 28 g/L (2.8% w/v), 35 g/L (3.5% w/v), 45 g/L (4.5% w/v), and 75 g/L (7.5% w/v).

[0397] Hypotonic saline solutions that are particularly useful for the methods and formulations described herein include those having a concentration from about 0.12 g/L (0.012% w/v) to about 8.5 g/L (0.85% w/v). Any concentration within this range may be used, such as 0.05%, 0.1%, 0.15%, 0.2%, 0.225% ( NS), 0.25%, 0.3% ( NS), 0.35%, 0.4%, 0.45% ( NS), 0.5%, 0.55%, 0.6% ( NS), 0.65%, 0.675% ( NS), 0.7%, 0.75%, or 0.8% w/v.

[0398] Each of the ranges and specific concentrations of saline described herein may be used with the formulations, uses/methods of treatment, regimens, and kits described herein.

[0399] Suitable anti-inflammatory agents for use in combination with the compounds of the disclosure include corticosteroids and non-steroidal anti-inflammatory drugs (NSAIDs), particularly phosphodiesterase (PDE) inhibitors. Examples of corticosteroids for use in the present disclosure include oral or inhaled corticosteroids or prodrugs thereof. Specific examples include but are not limited to ciclesonide, desisobutyryl-ciclesonide, budesonide, flunisolide, mometasone and esters thereof (e.g., mometasone furoate), fluticasone propionate, fluticasone furoate, beclomethasone, methyl prednisolone, prednisolone, dexamethasone, 6,9-difluoro-17-[(2-furanylcarbonyl)oxy]-11-hydroxy-16-methyl-3-oxo-androsta-1,4-diene-17-carbothioic acid S-fluoromethyl ester, 6,9-difluoro-11-hydroxy-16-methyl-3-oxo-17-propionyloxy-androsta-1,4-diene-17-carbothioic acid S-(2-oxo-tetrahydro-furan-3S-yl) ester, beclomethasone esters (e.g., the 17-propionate ester or the 17,21-dipropionate ester, fluoromethyl ester, triamcinolone acetonide, rofleponide, or any combination or subset thereof. Preferred corticosteroids for formulations, uses/methods of treatment, regimens, and kits disclosed herein are selected from ciclesonide, desisobutyryl-ciclesonide, budesonide, mometasone, fluticasone propionate, and fluticasone furoate, or any combination or subset thereof.

[0400] NSAIDs for use in the present disclosure include but are not limited to sodium cromoglycate, nedocromil sodium, phosphodiesterase (PDE) inhibitors (e.g., theophylline, aminophylline, PDE4 inhibitors, mixed PDE3/PDE4 inhibitors or mixed PDE4/PDE7 inhibitors), leukotriene antagonists, inhibitors of leukotriene synthesis (e.g., 5 LO and FLAP inhibitors), nitric oxide synthase (iNOS) inhibitors, protease inhibitors (e.g., tryptase inhibitors, neutrophil elastase inhibitors, and metalloprotease inhibitors), 02-integrin antagonists, adenosine receptor agonists or antagonists (e.g., adenosine 2a agonists), cytokine antagonists (e.g., chemokine antagonists) or inhibitors of cytokine synthesis (e.g., prostaglandin D2 (CRTh2) receptor antagonists). Examples of leukotriene modifiers suitable for administration by a method described herein include montelukast, zileuton and zafirlukast.

[0401] The PDE4 inhibitor, mixed PDE3/PDE4 inhibitor, or mixed PDE4/PDE7 inhibitor may be any compound that is known to inhibit the PDE4 enzyme or which is discovered to act as a PDE4 inhibitor, and which are selective PDE4 inhibitors (i.e., compounds which do not appreciably inhibit other members of the PDE family). Examples of PDE4 inhibitors for use in combination with the compounds disclosed herein include but are not limited to roflumilast, pumafentrine, arofylline, cilomilast, tofimilast, oglemilast, tolafentrine, piclamilast, ibudilast, apremilast, 2-[4-[6,7-diethoxy-2,3-bis(hydroxymethyl)-1-naphthalenyl]-2-pyridinyl]-4-(3-pyridinyl)-1(2H)-phthalazinone (T2585), N-(3,5-dichloro-4-pyridinyl)-1-[(4-fluorophenyl)methyl]-5-hydroxy--oxo-1H-indole-3-acetamide (AWD-12-281), 4-[(2R)-2-[3-(cyclopentyloxy)-4-methoxyphenyl]-2-phenylethyl]-pyridine (CDP-840), 2-[4-[[[[2-(1,3-benzodioxol-5-yloxy)-3-pyridinyl]carbonyl]amino]methyl]-3-fluorophenoxy]-(2R)-propanoic acid (CP-671305), N-(4,6-dimethyl-2-pyrimidinyl)-4-[4,5,6,7-tetrahydro-2-(4-methoxy-3-methylphenyl)-5-(4-methyl-1-piperazinyl)-1H-indol-1-yl]-benzenesulfonamide (2E)-2-butenedioate (YM-393059), 9-[(2-fluorophenyl)methyl]-N-methyl-2-(trifluoromethyl)-9H-purin-6-amine (NCS-613), N-(2,5-dichloro-3-pyridinyl)-8-methoxy-5-quinolinecarboxamide (D-4418), N-[(3R)-9-amino-3,4,6,7-tetrahydro-4-oxo-1-phenylpyrrolo[3,2,1-][1,4]benzodiazepin-3-yl]-3H-purin-6-amine (PD-168787), 3-[[3-(cyclopentyloxy)-4-methoxyphenyl]methyl]-N-ethyl-8-(1-methylethyl)-3H-purin-6-amine hydrochloride (V-11294A), N-(3,5-dichloro-1-oxido-4-pyridinyl)-8-methoxy-2-(trifluoromethyl)-5-quinolinecarboxamide (Sch351591), 5-[3-(cyclopentyloxy)-4-methoxyphenyl]-3-[(3-methylphenyl)methyl]-(3S,5S)-2-piperidinone (HT-0712), 5-(2-((1R,4R)-4-amino-1-(3-(cyclopentyloxy)-4-methyoxyphenyl)cyclohexyl) ethynyl)-pyrimidine-2-amine,cis-[4-cyano-4-(3-cyclopropylmethoxy-4-difluoromethoxy phenyl)cyclohexan-1-ol], and 4-[6,7-diethoxy-2,3-bis(hydroxymethyl)-1-naphthalenyl]-1-(2-methoxyethyl)-2(1H)-pyridinone (T-440), and any combination or subset thereof.

[0402] Leukotriene antagonists and inhibitors of leukotriene synthesis include zafirlukast, montelukast sodium, zileuton, and pranlukast.

[0403] Anticholinergic agents for use in combination with the compounds disclosed herein include but are not limited to muscarinic receptor antagonists, particularly including pan antagonists, and antagonists of the M.sub.3 receptors. Exemplary compounds include ipratropium, glycopyrrolate, tiotropium, the alkaloids of the belladonna plants, such as atropine, scopolamine, homatropine, hyoscyamine, and the various forms including salts thereof (e.g., anhydrous atropine, atropine sulfate, atropine oxide or HCl, methylatropine nitrate, homatropine hydrobromide, homatropine methyl bromide, hyoscyamine hydrobromide, hyoscyamine sulfate, scopolamine hydrobromide, scopolamine methyl bromide), or any combination or subset thereof. Additional anticholinergics for use in combination with the compounds disclosed herein include but are not limited to methantheline, propantheline bromide, anisotropine methyl bromide, Valpin 50, aclidinium bromide, umeclidinium bromide, glycopyrrolate (Robinul), isopropamide iodide, mepenzolate bromide, tridihexethyl chloride, hexocyclium methylsulfate, cyclopentolate HCl, tropicamide, trihexyphenidyl CCl, pirenzepine, telenzepine, and methoctramine, or any combination or subset thereof. Preferred anticholinergics for use in combination with the compounds disclosed herein include ipratropium (bromide), oxitropium (bromide), and tiotropium (bromide), or any combination or subset thereof.

[0404] Examples of -agonists for use in combination with the compounds disclosed herein include but are not limited to salmeterol and xinafoate salts thereof, R-salmeterol and xinafoate salts thereof, albuterol or R-albuterol (free base or sulfate), levalbuterol, salbutamol, formoterol (fumarate), fenoterol, procaterol, pirbuterol, metaprterenol, vilanterol, olodaterol, and terbutaline, or salts thereof, and any combination or subset thereof.

[0405] Examples of cystic fibrosis transmembrane conductance regulator (CFTR) modulators for use in combination with the compounds disclosed herein include but are not limited to CFTR potentiators, CFTR correctors, and CFTR amplifiers, and any combination or subset thereof, including ivacaftor, lumacaftor/ivacaftor, tezacaftor/ivacaftor, and elexacaftor/tezacaftor/ivacaftor. CFTR activity modulating compounds that can be administered in combination with a compound disclosed herein include, but are not limited to, compounds described in US 2009/0246137 A1, US 2009/0253736 A1, US 2010/0227888 A1, U.S. Pat. No. 7,645,789, US 2009/0246820 A1, US 2009/0221597 A1, US 2010/0184739 A1, US 2010/0130547 A1, US 2010/0168094 A1, U.S. Pat. Nos. 7,553,855, 7,772,259 B2, 7,405,233 B2, US. Pat. No. 2009/0203752, and U.S. Pat. No. 7,499,570, each of which are incorporated herein by reference.

[0406] P2Y2 receptor agonists for use in combination with the compounds disclosed herein may be employed in an amount effective to stimulate chloride and water secretion by airway surfaces, particularly nasal airway surfaces. P2Y2 agonists for use in combination with the compounds disclosed herein include P2Y2 receptor agonists such as ATP, UTP, UTP--S and dinucleotide P2Y2 receptor agonists (e.g. denufosol or diquafosol) or a pharmaceutically acceptable salt thereof. Suitable P2Y2 receptor agonists are known in the art and are described for example, in columns 9-10 of U.S. Pat. No. 6,264,975, and also U.S. Pat. Nos. 5,656,256 and 5,292,498. Suitable P2Y2 receptor agonists are described in, but are not limited to, U.S. Pat. Nos. 6,264,975, 5,656,256, 5,292,498, 6,348,589, 6,818,629, 6,977,246, 7,223,744, 7,531,525, and U.S. Pat. Pub. 2009/0306009, each of which is incorporated herein by reference.

[0407] P2Y14 antagonists for use in combination with the compounds disclosed herein include naphthoic acid and derivatives thereof (e.g., a substituted 2-naphthoic acid, 4-((piperidin-4-yl)-phenyl)-(7-(4-(trifluoromethyl)-phenyl)-2-naphthoic acid (PPTN)), and nucleotides and derivatives thereof (e.g., sugar nucleotides, uridine diphosphate).

[0408] Peroxisome proliferator-activated receptor (PPAR) agonists for use in combination with the compounds disclosed herein include compounds that modulate one or more of PPAR-alpha, -gamma, or -delta. Examples of PPAR agonists include, but are not limited to clofibrate, gemfibrozil, ciprofibrate, bezafibrate, fenofibrate, thiazolidinediones, NSAIDs, GW501516, oleoylethanolamide, palmitoylethanolamide, WY14643, pioglitazone, rosiglitazone, and ciglitazone.

[0409] Mucus or mucin modifying agents useful in the combinations and methods herein include reducing agents, surfactants and detergents, expectorants, and deoxyribonuclease agents.

[0410] Mucin proteins are organized into high molecular weight polymers via the formation of covalent (disulfide) and non-covalent bonds. Disruption of the covalent bonds with reducing agents is a well-established method to reduce the viscoelastic properties of mucus in vitro and is predicted to minimize mucus adhesiveness and improve clearance in vivo. Reducing agents are well known to decrease mucus viscosity in vitro and are commonly used as an aid to processing sputum samples. Examples of reducing agents include sulfide containing molecules or phosphines capable of reducing protein di-sulfide bonds including, but not limited to, N-acetyl cysteine (NAC), N-acystelyn, carbocysteine, glutathione, dithiothreitol, thioredoxin containing proteins, and tris (2-carboxyethyl) phosphine. Reducing agents such as NAC are not well suited for bolus aerosol administration. However, it is anticipated that delivery of reducing agents by pulmonary aerosol infusion would increase the effectiveness, while allowing for a decrease in the concentration of reducing agent in the inhalation solution (predicted to increase tolerability).

[0411] Mucoactive agents for use in combination with the compounds disclosed herein include expectorants, mucolytics, mucoregulators, and mucokinetics. Exemplary mucoactive agents include hypertonic saline, iodide-containing compounds, glyceryl guaiacolate (guaifenesin), ion channel modifiers, anticholinergic agents, glucocorticoids, prednisolone, iodinated glycerol, domiodol, tricyclic nucleotides (e.g., uridine triphosphate and adenosine triphosphate), sodium citrate, potassium citrate, carbocysteine, potassium iodide, guaifenesin, tolu balsam, vasaka, ammonium chloride, macrolide antibiotics (e.g., erythromycin, azithromycin, clarithromycin, roxithromycin), acetylcysteine, acystelyn, ambroxol, bromhexine, carbocisteine, erdosteine, fudosteine, mecysteine, gelsolin, thymosin 04, non-destructive mucolytics (e.g., dextran and heparin), bronchodilators, tricyclic nucleotides, broxol and dornase alfa, HCl, or any combination or subset thereof.

[0412] Surfactants and detergents are spreading agents shown to decrease mucus viscoelasticity, thus improving mucus clearability. Examples of surfactants include dipalmitoyl phosphatidylcholine (DPPC), palmitic acid, palmitoyl-oleoylphosphatidylglycerol, surfactant-associated proteins (e.g. SP-A, B, or C), or animal derived (e.g. from cow or calf lung lavage or extracted from minced pig lung) or combinations thereof. See, e.g., U.S. Pat. Nos. 7,897,577, 5,876,970, 5,614,216, 5,100,806, and 4,312,860. Examples of surfactant products include colfosceril palmitate, DPPC and egg phosphatidylglycerol, KL-4 surfactant, lusulptide, rSP-C surfactant, bovactant, poractant alfa, calfactant, modified bovine surfactant, Surface, nonionic alcohol ethoxylate surfactant, and beractant. Examples of detergents include, but are not limited to, Tween-80 and triton-X 100.

[0413] Any suitable expectorant can be used, including but not limited to guaifenesin (see, e.g., U.S. Pat. No. 7,345,051). Any suitable deoxyribonuclease can be used, including but not limited to Dornase Alpha. (see, e.g., U.S. Pat. No. 7,482,024).

[0414] Examples of kinase inhibitors for use in combination with the compounds disclosed include inhibitors of NFkB, PI3K (phosphatidylinositol 3-kinase), p38-MAP kinase, and Rho kinase.

[0415] Hydrating agents for use in combination with the compounds disclosed herein include ivacaftor, hypertonic saline, mannitol, lumacaftor, amiloride, amiloride analogs, camostat, denufosol, duramycin, 3-isobutyl-1methylxantine, betaine, bortezomib, velcade, bisaminomethylbithizaole, curcumin, geneticin, genistein, gentamicin, glycerol, matrine, miglustat, ataluren, sildenafil, sildenafil analogs, buphenyl, suberoylanilide hydroxamic acid, thapsigargin, tobramycin, trimethylamine N-oxide, ivacaftor, lumacaftor, VRT-325, VRT-532, and INO-4995.

[0416] Examples of immune-modulatory agents for use in combination with the compounds disclosed included calcineurin inhibitors (e.g., cyclosporine), antimetabolites (e.g., purine analogues (e.g., azathioprine and mycophenolate mofetil) and folate antagonists (e.g., methotrexate and dapsone), and alkylating agents (e.g., cyclophosphamide).

[0417] Antiinfective agents for use in combination with the compounds disclosed herein include antivirals and antibiotics. Examples of suitable antivirals include Tamiflu (oseltamivir) and Relenza (zanamivir). Examples of suitable antibiotics include but are not limited to aztreonam (arginine or lysine), fosfomycin, and aminoglycosides such as tobramycin, or any combination or subset thereof. In certain embodiments, the additional therapeutically agent is an antibiotic. Exemplary antibiotics include, but are not limited to, penicillins (e.g., penicillin, amoxicillin), cephalosporins (e.g., cephalexin), macrolides (e.g., erythromycin, clarithromycin, azithromycin, troleandomycin), fluoroquinolones (e.g., ciprofloxacin, levofloxacin, ofloxacin, delafloxacin), sulfonamides (e.g., co-trimoxazole, trimethoprim), tetracyclines (e.g., tetracycline, chlortetracycline, oxytetracycline, demeclocycline, methacycline, sancycline, doxycline, aureomycin, terramycin, minocycline, 6-deoxytetracycline, lymecycline, meclocycline, methacycline, rolitetracycline, and glycylcycline antibiotics (e.g., tigecycline)), aminoglycosides (e.g., gentamicin, tobramycin, paromomycin), aminocyclitol (e.g., spectinomycin), chloramphenicol, sparsomycin, and quinupristin/dalfoprisin. Additional antiinfective agents that may be used herein include aminoglycosides, daptomycin, fluoroquinolones, ketolides, carbapenems, cephalosporins, erythromycin, linezolid, penicillins, azithromycin, clindamycin, oxazolidinones, tetracyclines, and vancomycin. Examples of useful carbapenem antibiotics are imipenem, panipenem, meropenem, biapenem, MK-826 (L-749,345), DA-1131, ER-35786, lenapenem, S-4661, CS-834 (prodrug of R-95867), KR-21056 (prodrug of KR-21012), L-084 (prodrug of LJC 11036), and Ceftolozane (CXA-101).

[0418] Examples of other classes of therapeutic agents suitable for use in a combination herein include antivirals such as ribavirin, anti-fungal agents such as amphotericin, itraconazole and voriconazole, anti-rejection drugs such as cyclosporine, tacrolimus and sirolimus, bronchodilators including but not limited to anticholinergic agents such as atrovent, siRNAs, gene therapy vectors, aptamers, endothelin-receptor antagonists, alpha-1-antitrypsin and prostacyclins.

[0419] Antihistamines (i.e., H1-receptor antagonists) for use in combination with the compounds disclosed herein include but are not limited to: ethanolamines, diphenhydramine HCl, carbinoxamine maleate, doxylamine, clemastine fumarate, diphenylhydramine HCl. dimenhydrinate, ethylenediamines, pyrilamine maleate (metpyramine), tripelennamine HCl, tripelennamine citrate, antazoline, alkylamines, pheniramine, chloropheniramine, bromopheniramine, dexchlorpheniramine, triprolidine, acrivastine, pyridines methapyrilene, piperazines, hydroxyzine HCl, hydroxyzine pamoate, cyclizine HCl, cyclizine lactate, meclizine HCl, cetirizine HCl, piperidines, astemisole, levocabastine HCl, loratadine, descarboethoxyloratadine, terfenadine, fexofenadine HCl, tri- and tetracyclics, promethazine, chlorpromethazine trimeprazine, azatadine, and azelastine HCl, or any combination or subset thereof.

[0420] Combination therapies herein can include adenosine 2b (A2b) agonists, including BAY 60-6583, NECA (N-ethylcarboxamidoadenosine), (S)-PHPNECA, LUF-5835 and LUF-5845. A2b agonists that may be used are described by Volpini et al., Journal of Medicinal Chemistry 45 (15): 3271-9 (2002); Volpini et al., Current Pharmaceutical Design 8 (26): 2285-98 (2002); Baraldi et al., Journal of Medicinal Chemistry 47 (6): Cacciari et al., 1434-47 (2004); Mini Reviews in Medicinal Chemistry 5 (12): 1053-60 (December 2005); Baraldi et al., Current Medicinal Chemistry 13 (28): 3467-82 (2006); Beukers et al., Medicinal Research Reviews 26 (5): 667-98 (September 2006); Elzein et al., Bioorganic & Medicinal Chemistry Letters 16 (2): 302-6 (January 2006); Carotti, et al., Journal of Medicinal Chemistry 49 (1): 282-99 (January 2006); Tabrizi et al., Bioorganic & Medicinal Chemistry 16 (5): 2419-30 (March 2008); and Stefanachi, et al., Bioorganic & Medicinal Chemistry 16 (6): 2852-69 (March 2008).

[0421] Examples of other ENaC receptor blockers for use in combination with the compounds of the disclosure include but are not limited to amiloride and derivatives thereof such as those compounds described in U.S. Pat. No. 6,858,615, and PCT Publication Nos. WO2003/070182, WO2004/073629, WO2005/018644, WO2006/022935, WO2007/018640, and WO2007/146869, all to Parion Sciences, Inc.

[0422] Small molecule ENaC blockers are capable of directly preventing sodium transport through the ENaC channel pore. ENaC blocker that can be administered in the combinations herein include, but are not limited to, amiloride, benzamil, phenamil, and amiloride analogues as exemplified by U.S. Pat. Nos. 6,858,614, 6,858,615, 6,903,105, 6,995,160, 7,026,325, 7,030,117, 7,064,129, 7,186,833, 7,189,719, 7,192,958, 7,192,959, 7,241,766, 7,247,636, 7,247,637, 7,317,013, 7,332,496, 7,345,044, 7,368,447, 7,368,450, 7,368,451, 7,375,107, 7,399,766, 7,410,968, 7,820,678, 7,842,697, 7,868,010, and 7,875,619.

[0423] ENaC proteolysis is well described to increase sodium transport through ENaC. Protease inhibitors block the activity of endogenous airway proteases, thereby preventing ENaC cleavage and activation. Proteases that cleave ENaC include furin, meprin, matriptase, trypsin, channel associated proteases (CAPs), and neutrophil elastases. Protease inhibitors that can inhibit the proteolytic activity of these proteases that can be administered in the combinations herein include, but are not limited to, camostat, prostasin, furin, aprotinin, leupeptin, and trypsin inhibitors.

[0424] Combinations herein may include one or more suitable nucleic acid (or polynucleic acid), including but not limited to antisense oligonucleotide, siRNA, miRNA, miRNA mimic, antagomir, ribozyme, aptamer, and decoy oligonucleotide nucleic acids. See, e.g., US Patent Application Publication No. 20100316628. In general, such nucleic acids may be from 17 or 19 nucleotides in length, up to 23, 25 or 27 nucleotides in length, or more. Examples include, but are not limited to, those described in U.S. Pat. No. 7,517,865 and US Patent Publication Applications Nos. 20100215588, 20100316628, 20110008366, and 20110104255. In general, the siRNAs are from 17 or 19 nucleotides in length, up to 23, 25 or 27 nucleotides in length, or more. In some embodiments, the siRNA is ARO-ENAC or IONIS ENACR-x.

[0425] In some embodiments, the combinations described herein may be present in either a single or multiple compositions. In some embodiments, the combinations described herein are present in either a single composition comprising a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, with one or more other therapeutically active agents. In some embodiments, the combinations described herein are present in more than one composition wherein one composition comprises a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, and a second composition comprises one or more other therapeutically active agents.

[0426] In one preferred embodiment, a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is used in combination with one or more osmolytes. In one embodiment, a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is used in combination with hypertonic saline. In some embodiments, the compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, and hypertonic saline are delivered as an aerosol formulation. In one embodiment, a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is used in combination with mannitol. In one embodiment, a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is used in combination with xylitol. In one embodiment, a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is used in combination with xylitol and hypertonic saline. In some embodiments, the compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, xylitol, and hypertonic saline is delivered as an aerosol formulation. In one embodiment, a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is used in combination with sodium gluconate.

[0427] In another preferred embodiment, a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is used in combination with one or more osmolytes and an excipient (i.e., used as a pharmaceutical composition). In another preferred embodiment, a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, is used in combination with hypertonic saline and a cyclodextrin. In some embodiments, the compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled derivative, solvate, hydrate, polymorph, co-crystal, or prodrug thereof, hypertonic saline, and the cyclodextrin is delivered as an aerosol formulation.

EXAMPLES

[0428] The present invention also provides processes for preparing the compounds of the invention and to the synthetic intermediates useful in such processes, as described in detail below.

[0429] Certain abbreviations and acronyms are used in describing the synthetic processes and experimental details. Although most of these would be understood by one skilled in the art, the following table contains a list of many of these abbreviations and acronyms.

[0430] The following abbreviations are used above and hereinafter:

Abbreviation Meaning

[0431] AcOH Acetic Acid [0432] ACN Acetonitrile [0433] Aq aqueous [0434] AUC Area under the curve [0435] BOC tert-Butoxycarbonyl [0436] Cbz Carbobenzyloxy [0437] CH Cyclohexane [0438] DCE Dichloroethane [0439] DCM Dichloromethane [0440] DIAD Diisopropyl azodicarboxylate [0441] DIPEA Diisopropylethylamine [0442] DMAP 4-Dimethylaminopyridine [0443] DMF N,N-Dimethylformamide [0444] Dppf 1, 1-Bis(diphenylphosphino)ferrocene [0445] EDCIEthyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride [0446] ENaCi Epithelial Sodium Channel Inhibitor [0447] Et Ethyl [0448] EtOAc or EA Ethyl acetate [0449] EtOH ethanol [0450] Equiv Molar equivalent [0451] ESI Electrospray ionization [0452] h hour [0453] HATU 0-(7-Azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate [0454] HCl Hydrochloric acid HPLC High performance liquid chromatography [0455] IPA or iPrOH Isopropyl Alcohol [0456] KOH Potassium hydroxide [0457] L liter [0458] LCMS liquid chromatography mass spectroscopy [0459] LiHMDS Lithium bis(trimethylsilyl)amide [0460] M mol/1 [0461] Me methyl [0462] MeOH Methanol [0463] Min minutes [0464] mL milliliter [0465] Mp melting point m/z [0466] NaOH Sodium hydroxide [0467] m/z mass to charge ratio [0468] MH.sup.+ mass plus one [0469] MH mass minus one [0470] MHz megahertz [0471] MS or ms Mass spectrum [0472] MTBE methyl tertiary butyl ether [0473] n.d. not determined [0474] NMP N-Methylpyrrolidone [0475] Pd/C palladium on charcoal [0476] Ph.sub.3P triphenyl phosphine [0477] t-Bu tert-butyl [0478] r.t. or rt ambient temperature (about 20 C.) [0479] t.sub.R retention time [0480] MTBE Methyl tert-butyl ether [0481] TEA Triethylamine [0482] TFA Trifluoroacetic acid [0483] THF Tetrahydrofuran [0484] TLC Thin Layer Chromatography [0485] TMS Trimethylsilyl [0486] UPLC Ultra Performance Liquid Chromatography [0487] UV Ultraviolet [0488] wt % Percent by weight

[0489] The compounds described herein may be synthesized using techniques and procedures known in the art. These procedures are described in, for example, E. J. Cragoe, The Synthesis of Amoloride and Its Analogs (Chap 3) in Amiloride and its Analogs, pp 25-36. Other processes for preparing amiloride analogues are described in. for example, U.S. Pat. No. 3,318,813, to Cragoe, particularly at methods A, B, C, and D of the '813 patent. Still other processes which may be adapted for the preparation of the compounds of the invention are described in U.S. Pat. Nos. 6,858,615, 6,903,105, 7,064,129, 7,399,766, 8,669,262, 9,029,382, 9,102,633, and 9,593,084.

[0490] Methyl N-3,5-diamino-6-chloropyrazine-2-carbonylcarbamimido thioate (compound 1) is commercially available and also can be prepared as seen in WO2011/156355 A1.

[0491] As will be apparent to those skilled in the art, in certain instances, the starting or intermediate compounds in the synthesis may possess other functional groups which provide alternate reactive sites. Interference with such functional groups may be avoided by utilization of appropriate protecting groups such as amine or alcohol protecting groups and where applicable appropriately prioritizing the synthetic steps. Suitable protecting groups will be apparent to those skilled in the art. Methods are well known in the art for installing and removing such protecting groups and such conventional techniques may be employed in the processes of the instant inventions as well.

Synthetic Schemes

[0492] Some exemplary synthetic schemes that may be used to synthesize the compounds described herein appear below:

##STR00300##

##STR00301##

[0493] In some embodiments, M is a metal. In some embodiments, M is B, Mg, Fe, or Zn. In some embodiments, LG, the leaving group is halogen or OTf. In some embodiments, LG is Cl, Br, I, or OTf.

##STR00302##

[0494] In some embodiments, M is a metal. In some embodiments, M is B, Mg, Fe, or Zn. In some embodiments, LG, the leaving group is halogen or OTf. In some embodiments, LG is Cl, Br, I, or OTf.

##STR00303##

##STR00304##

##STR00305##

##STR00306##

Materials and Methods

[0495] All commercial materials were used as supplied unless otherwise noted. All solvents were reagent grade or HPLC grade. Anhydrous DMF, THF, EtOH, 1,4-dioxane, MeOH, CH.sub.2Cl.sub.2 were purchased from Sigma-Aldrich and used without further drying. All reactions were performed under an atmosphere of pre-purified dry Ar(g) unless specified otherwise. NMR spectra were recorded on Bruker Avance-300 and Bruker Avance-500 instrument. NMR Solvents CDCl.sub.3, CD.sub.3OD and DMSO-d.sub.6 were purchased from Aldrich or Cambridge Isotope Laboratories, unless otherwise specified. The following abbreviations were used to explain the multiplicities: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, and br=broad. Chemical shifts are reported in ppm relative to tetramethylsilane (TMS) as the internal standard. All reactions were carried out in oven-dried glassware under argon atmosphere unless otherwise noted. Reactions were monitored by TLC carried out on 0.25 mm E. Merck silica-gel plates (60F-254) by using UV light as visualizing agent and ninhydrin solution and heat as developing agents. For polar compounds reactions are monitored by HPLC and LCMS analysis. RediSep Rf Gold Silica Gel column (40-60 microns) was used for flash-column chromatography. Reusable RediSep Rf C18 reverse phase columns (40-60 microns) was used for reverse phase chromatography.

[0496] The following HPLC/UPLC methods were used to characterize and monitor intermediates and examples. Column, wavelength, flow rates, and solvent gradients are given in the individual methods.

HPLC Methods

HPLC Method A

TABLE-US-00002 Column: YMC ODS-AQ Column, 150 4.6 mm Detector: UV: 254 nm Gradient time % Sol (ACN % Sol (H.sub.2O Flow (min) w/0.1% TFA) w/0.1%TFA) (mL/min) 0.00 5 95 1 15.00 100 0 1 19.00 100 0 1

UPLC Method B

TABLE-US-00003 Column: UPLC BEH C18 Column, 75 2.1 mm, 1.7 m Detector: UV: 254 nm Gradient time % Sol (ACN % Sol (H.sub.2O Flow (min) w/0.1% TFA) w/0.1%TFA) (mL/min) 0.00 5 95 0.5 6 0 100 0.5 8 0 100 0.5 9 95 5 0.5

HPLC Method C

TABLE-US-00004 Column: Xbridge C18 Column, 150 4.6 mm; 5.0 m Detector: UV: 220 nm Gradient % Sol (H.sub.2O; ACN; % Sol (H.sub.2O; ACN; time 9:1; w/50 mM 1:9; w/50 mM Flow (min) ammonium formate) ammonium formate) (mL/min) 0.00 95 5 1 3.00 95 5 1 10.00 10 90 1 16.00 10 90 1 16.50 95 5 1 20.00 95 5 1

HPLC Method D

TABLE-US-00005 Column: Xbridge C18 Column, 150 4.6 mm; 3.5 m Detector: UV: 220 nm Gradient time % Sol (H.sub.2O % Sol (ACN Flow (min) w/0.05% TFA) w/0.05% TFA) (mL/min) 0.00 95 5 1 5.00 95 5 1 15.00 10 90 1 20.00 10 90 1 20.10 95 5 1 25.00 95 5 1

HPLC Method E

TABLE-US-00006 Column: Xbridge C18 Column, 150 4.6 mm; 3.5 m Detector: UV: 220 nm Gradient % Sol (H.sub.2O % Sol (ACN time w/0.05% w/0.05% Flow (min) TFA) TFA) (mL/min) 0.00 95 5 1 3.00 95 5 1 6.00 10 90 1 12.00 10 90 1 12.10 95 5 1 16.00 95 5 1

HPLC Method F

TABLE-US-00007 Column: Zodiac C18 Column, 100 4.6 mm; 3.0 m Detector: UV: 254 nm Gradient time % Sol (H.sub.2O % Sol (ACN Flow (min) w/0.05% TFA) w/0.05% TFA) (mL/min) 0.00 95 5 1 3.00 95 5 1 6.00 10 90 1 12.00 10 90 1 12.10 95 5 1 15.00 95 5 1

HPLC Method G

TABLE-US-00008 Column: Zodiac C18 Column, 100 4.6 mm; 3.0 m Detector: UV: 220 nm Gradient time % Sol (H.sub.2O % Sol (ACN Flow (min) w/0.05% TFA) w/0.05% TFA) (mL/min) 0.00 95 5 0.8 3.00 95 5 0.8 6.00 10 90 0.8 12.00 10 90 0.8 12.10 95 5 0.8 15.00 95 5 0.8

HPLC Method H

TABLE-US-00009 Column: Zodiac C18 Column, 150 4.6 mm; 5.0 m Detector: UV: 254 nm % Sol H.sub.2O Gradient w/10 mM time ammonium % Sol Flow (min) formate ACN (mL/min) 0.00 80 20 1 5.00 80 20 1 15.00 10 90 1 20.00 10 90 1 20.10 80 20 1 25.00 80 20 1

[0497] HPLC Method I

TABLE-US-00010 Column: Polaris C18-A Column, 100 3.0 mm; 2.6 m Detector: UV: 254 nm Gradient % Sol (H.sub.2O % Sol (ACN Flow time (min) w/0.05% TFA) w/0.05% TFA) (mL/min) 0.00 95 5 0.8 3.00 95 5 0.8 6.00 10 90 0.8 12.00 10 90 0.8 12.10 95 5 0.8 15.00 95 5 0.8

HPLC Method J

TABLE-US-00011 Column: Polaris C18-A Column, 100 3.0 mm; 2.6 m Detector: UV: 220 nm Gradient % Sol (H.sub.2O % Sol (ACN Flow time (min) w/0.05% TFA) w/0.05% TFA) (mL/min) 0.00 95 5 0.8 3.00 95 5 0.8 6.00 10 90 0.8 12.00 10 90 0.8 12.10 95 5 0.8 15.00 95 5 0.8

HPLC Method K

TABLE-US-00012 Column: Polaris C18 Column, 100 3.0 mm; 2.6 m Detector: UV: 215 nm Gradient % Sol (H.sub.2O % Sol (ACN Flow time (min) w/0.05% TFA) w/0.05% TFA) (mL/min) 0.00 95 5 0.8 3.00 95 5 0.8 6.00 10 90 0.8 12.00 10 90 0.8 12.10 95 5 0.8 15.00 95 5 0.8

HPLC Method L

TABLE-US-00013 Column: Polaris C18 Column, 100 3.0 mm; 2.6 m Detector: UV: 220 nm Gradient % Sol (H.sub.2O % Sol (ACN Flow time (min) w/0.05% TFA) w/0.05% TFA) (mL/min) 0.00 95 5 0.8 4.00 95 5 0.8 8.00 10 90 0.8 15.00 10 90 0.8 15.10 95 5 0.8 20.00 95 5 0.8

HPLC Method M

TABLE-US-00014 Column: Polaris C18 Column, 100 3.0 mm; 2.6 m Detector: UV: 215 nm Gradient % Sol (H.sub.2O % Sol (ACN Flow time (min) w/0.05% TFA) w/0.05% TFA) (mL/min) 0.00 95 5 0.8 5.00 95 5 0.8 10.00 10 90 0.8 20.00 10 90 0.8 20.10 95 5 0.8 25.00 95 5 0.8

HPLC Method N

TABLE-US-00015 Column: Polaris C18 Column, 100 3.0 mm; 2.6 m Detector: UV: 220 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 0.8 5.00 95 5 0.8 10.00 10 90 0.8 20.00 10 90 0.8 20.10 95 5 0.8 25.00 95 5 0.8

HPLC Method O

TABLE-US-00016 Column: Eclipse Plus C18 Column, 100 4.6 mm; 3.5 m Detector: UV: 260 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 1 3.00 95 5 1 5.00 30 70 1 10.00 5 95 1 12.00 5 95 1 12.10 95 5 1 15.00 95 5 1

HPLC Method P

TABLE-US-00017 Column: Eclipse Plus C18 Column, 100 4.6 mm; 3.5 m Detector: UV: 215 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 1 3.00 95 5 1 5.00 30 70 1 10.00 5 95 1 12.00 5 95 1 12.10 95 5 1 15.00 95 5 1

HPLC Method Q

TABLE-US-00018 Column: Eclipse Plus C18 Column, 100 4.6 mm; 3.5 m Detector: UV: 220 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 0.8 3.00 95 5 0.8 6.00 10 90 0.8 12.00 10 90 0.8 12.10 95 5 0.8 15.00 95 5 0.8

HPLC Method R

TABLE-US-00019 Column: Eclipse Plus C18 Column, 100 4.6 mm; 3.5 m Detector: UV: 254 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 0.8 3.00 95 5 0.8 6.00 10 90 0.8 12.00 10 90 0.8 12.10 95 5 0.8 15.00 95 5 0.8

HPLC Method S

TABLE-US-00020 Column: Eclipse Plus C18 Column, 100 4.6 mm; 3.5 m Detector: UV: 254 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 0.8 2.00 95 5 0.8 8.00 30 70 0.8 10.00 10 90 0.8 12.00 10 90 0.8 12.10 95 5 0.8 15.00 95 5 0.8

HPLC Method T

TABLE-US-00021 Column: Eclipse Plus C18 Column, 100 4.6 mm; 3.5 m Detector: UV: 265 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 0.8 2.00 95 5 0.8 8.00 30 70 0.8 10.00 5 95 0.8 12.00 5 95 0.8 12.10 95 5 0.8 15.00 95 5 0.8

HPLC Method U

TABLE-US-00022 Column: Eclipse Plus C18 Column, 100 4.6 mm; 3.5 m Detector: UV: 260 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 0.8 3.00 95 5 0.8 8.00 30 70 0.8 10.00 10 90 0.8 15.00 10 90 0.8 15.10 95 5 0.8 20.00 95 5 0.8

HPLC Method V

TABLE-US-00023 Column: Eclipse Plus C18 Column, 150 4.6 mm; 3.5 m Detector: UV: 260 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 0.8 3.00 95 5 0.8 8.00 30 70 0.8 10.00 10 90 0.8 15.00 10 90 0.8 15.10 95 5 0.8 20.00 95 5 0.8

HPLC Method W

TABLE-US-00024 Column: Eclipse Plus C18 Column, 100 4.6 mm; 3.5 m Detector: UV: 260 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 0.8 2.00 95 5 0.8 8.00 30 70 0.8 12.00 10 90 0.8 12.10 95 5 0.8 15.00 95 5 0.8

HPLC Method X

TABLE-US-00025 Column: Luna C18 Column, 150 4.6 mm; 3.0 m Detector: UV: 254 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 0.8 3.00 95 5 0.8 6.00 10 90 0.8 12.00 10 90 0.8 12.10 95 5 0.8 15.00 95 5 0.8

HPLC Method Y

TABLE-US-00026 Column: Luna C18 Column, 150 4.6 mm; 3.0 m Detector: UV: 254 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 0.8 5.00 95 5 0.8 10.00 10 90 0.8 15.00 10 90 0.8 15.10 95 5 0.8 20.00 95 5 0.8

HPLC Method Z

TABLE-US-00027 Column: Eclipse Plus C18 Column, 100 4.6 mm; 3.5 m Detector: UV: 215 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 95 5 1.0 2.00 95 5 1.0 7.00 30 70 1.0 12.00 5 95 1.0 12.10 95 5 1.0 15.00 95 5 1.0

UPLC Method AA

TABLE-US-00028 Column: Gemini NX-C18 Column, 4.6 100 mm, 3.0 m Detector: ELSD Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 98 2 0.8 6.00 5 95 0.8 8.00 98 2 0.8

UPLC Method AB

TABLE-US-00029 Column: XBridge_BEH C18 Column, 2.5 m (50 2.1 mm) Detector: UV: 215 nm Gradient % Sol (H.sub.2O % Sol (ACN time (min) w/0.05% TFA) w/0.05% TFA) Flow (mL/min) 0.00 90 10 0.6 1.50 90 10 0.6 5.00 30 70 0.6 7.00 5 95 0.6 8.60 90 10 0.6 10.00 90 10 0.6

[0498] The following specific examples which are provided herein for the purposes of illustration only and do not limit the scope of the invention, which is defined by the claims.

INTERMEDIATES

Intermediate 1

Preparation of 4-(4-(((benzyloxy)carbonyl)amino)butyl)phenyl trifluoromethanesulfonate (Int-1)

##STR00307##

[0499] Preparation of Int-1: To a stirred solution of Int-1a (100 g, 334 mmol) in pyridine (600 mL) was added triflic anhydride (67 mL, 401 mmol) at 0 C. Reaction mixture was stirred at 0 C. for 2 h. Upon completion the reaction mixture poured into a beaker containing cold water (300 mL) under stirring. Gummy material formed, which become solid after seeding with reference solid of compound Int-1 (ca. 500 mg). Stirred for 1 h and filtered the solid, washed with water (1000 mL) and dried in the Buckner flask under vacuum. The obtained solid was stirred in hexanes (2000 mL) and filtered to remove residual pyridine. The solid was dried under reduced pressure to afford compound Int-1 (130 g, 90%) as an off-white solid. ESI (m/z) [C.sub.19H.sub.20F.sub.3NO.sub.5S+H].sup.+ 432. HPLC purity 99.2%; HPLC method: columnPolaris C18-A 1003.0 mm, 2.6 m; 0.1% TFA in ACN:water (1:1) at 215 nm.

Intermediate 2

Preparation of 4-(4-((tert-butoxycarbonyl)amino)butyl)phenyl trifluoromethanesulfonate (Int-2)

##STR00308##

[0500] Preparation of Int-2a: A stirred solution of Int-1a (4.0 g, 16.7 mmol) in IPA (40 mL) and CH.sub.3COOH (0.5 mL), was charged with Pd(OH).sub.2/C (400 mg, 20 mol %) at room temperature. The resultant reaction mixture stirred under H.sub.2 balloon pressure for 2 h. (Boc).sub.2O (5.70 mL, 25.18 mmol) was added to the reaction mixture and stirred for 1 h. The obtained crude compound was partitioned between water (100 mL) and EtOAc (150 mL), and the separated organic layer was washed with brine solution (50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. Obtained crude was triturated in hexanes, filtered the precipitated solid and dried to afford Int-2a (3.20 g, 91%) as an off-white solid. ESI (m/z) [C.sub.15H.sub.23NO.sub.3+H].sup.+ 266.

[0501] Preparation of Int-2: To a stirred solution of Int-2a (3.20 g, 12.07 mmol) in pyridine (20 mL), was added triflic anhydride (3.70 g, 13.28 mmol) at 0 C. Reaction mixture was stirred at 0 C. for 1 h. Upon completion, the reaction mixture poured into a beaker containing cold water (50 mL) with stirring. The aqueous layer was extracted with EtOAc (250 mL), the combined organic layers were washed with 2N HCl solution (320 mL) followed by brine solution (50 mL), then dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford compound Int-2 (3.60 g, 77%) as an brown liquid. ESI (m/z) [C.sub.16H.sub.22F.sub.3NO.sub.5S+H].sup.+ 398.

Intermediate 3

Preparation of methyl 3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoate (Int-3)

##STR00309##

[0502] Preparation of Int-3: To a stirred solution of commercially available Int-3a (14.0 g, 57.6 mmol) and bis-pinacolato-diboron (17.50 g, 69.1 mmol) in 1,4-dioxane (150 mL) was added KOAc (17.90 g, 172.8 mmol) at room temperature and degassed for 10 min with Argon then added Pd(dppf).sub.2Cl.sub.2. The reaction mixture was heated to 100 C. and stirred for 6 h. After completion of the reaction, the mixture was cooled to room temperature and filtered through celite bed and washed with EtOAc (100 mL). The filtrate was concentrated under reduced pressure and the residue was purified by combiflash column (220 g column size) chromatography eluting with 5% EtOAc in hexanes to afford Int-3 (12.0 g, 70%) as an off-white solid. ESI (m/z) [C.sub.16H.sub.23BO.sub.4+H].sup.+ 291.

Intermediate 4-5

[0503] Preparation of Intermediates 4-5: In an analogous procedure used for preparation of Int-3, Intermediates 4-5 were synthesized from the corresponding commercially available phenyl bromides:

TABLE-US-00030 Int-# Structure [M + H].sup.+ 4 [00310] Carried forward without purification 5 [00311] 291

Intermediate 6

Preparation of methyl 3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanoate (Int-6)

##STR00312##

[0504] Preparation of Int-6a: To a stirred solution of commercially available (4-bromo-3-methylphenyl)methanol (7.30 g, 36.5 mmol) in CH.sub.2Cl.sub.2 (70 mL) was added Dess-Martin periodinane (23.20 g, 54.5 mmol) at 0 C. The reaction mixture was stirred for 1 h. After completion of the reaction, reaction mixture was quenched with NaHCO.sub.3 and hypo solution (10% aqueous Sodium thiosulfate (Na.sub.2S.sub.2O.sub.3)) (1:1, 200 mL) and extracted with CH.sub.2Cl.sub.2 (3100 mL). Combined organic layer was evaporated under reduced pressure to afford crude material and which was washed with hexanes to give desire compound Int-6a (7.0 g, 89%) as an off-white solid. ESI (m/z) [C.sub.8H.sub.7BrO+H].sup.+ 200.

[0505] Preparation of Int-6b: To a stirred solution of compound Int-6a (7.0 g, 35.5 mmol) in toluene (70 mL) was pre-heated to 60 C., then added (carbomethoxymethylene)triphenylphosphorane (23.60 g, 70.7 mmol) and stirred for 1 h at 70 C. After completion of the reaction, reaction mixture was evaporated under reduced pressure to get crude material. The obtained crude was purified by combi-flash (220 g column) chromatography eluting with 5% EtOAc in hexanes to afford compound Int-6b (7.0 g, 77%) as an off-white solid. ESI (m/z) [C.sub.11H.sub.11BrO.sub.2+H].sup.+ 256.

[0506] Preparation of Int-6c: To a stirred solution of compound Int-6c (6.60 g, 25.98 mmol) and bis-pinacolato-diboron (7.80 g, 31.18 mmol) in 1,4-dioxane (70 mL) was added KOAc (7.60 g, 77.94 mmol) at room temperature and degassed for 10 min with argon then added Pd(dppf)Cl.sub.2.Math.DCM and heated at 100 C. for 6 h. Reaction mixture was cooled to room temperature and filtered via celite bed and washed with EtOAc and concentrated. The obtained crude was purified by combiflash (80 g column) chromatography eluting with 5% EtOAc in hexanes to afford compound Int-6c (6.4 g, 82%) as an off-white solid. ESI (m/z) [C.sub.17H.sub.23BO.sub.4+H].sup.+ 303.

[0507] Preparation of Int-6: To a stirred solution of compound Int-6c (6.40 g, 57.6 mmol) in EtOH (100 mL) was added Pd/C (0.50 g), then stirred for 16 h under H.sub.2 balloon. After completion of the reaction, reaction mixture was filtered via celite bed, washed with EtOAc and concentrated under reduced pressure to afford desire compound Int-6 (6.0 g, 92%) as an off-white solid. ESI (m/z) [C.sub.17H.sub.25BO.sub.4+H].sup.+ 305.

Intermediate 7

Preparation of methyl (E)-3-(3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acrylate (Int-7)

##STR00313##

[0508] Preparation of Int-7a: A stirred solution of 4-bromo-3-chlorobenzaldehyde (1.0 g, 4.55 mmol) in toluene (30 mL) was charged methyl triphenylphosphoranylideneacetate (1.90 g, 5.46 mmol) at ambient temperature. The resultant reaction mixture was heated to 100 C. and stirred for 16 h. The reaction mixture was concentrated under reduced pressure and purified by combi-flash column chromatography eluting with (10-100% EtOAc in hexanes) to afford Int-7a (1.20 g, 91%) as a white solid. ESI (m/z) [C.sub.10H.sub.8BrClO.sub.2+H].sup.+ 275.

[0509] Preparation of Int-7: To a stirred solution of compound Int-7a (5.00 g, 18.1 mmol) and Bis(pinacolato)diboron (9.20 g, 36.3 mmol) in 1,4-dioxane (50 mL) was added KOAc (3.70 g, 54.5 mmol) at room temperature. The reaction mixture was degassed for 10 min with Argon then Pd(dppf).sub.2Cl.sub.2 (0.66 g, 0.90 mmol) added. The reaction mixture was warmed to 100 C. and stirred for 6 h. After completion of the reaction, the mixture was cooled to room temperature and filtered through celite bed and washed with MTBE (100 mL). The filtrate was concentrated under reduced pressure and obtained crude material was purified by combi-flash chromatography eluting with (1%-100% EtOAc in hexanes) to afford compound Int-7 (7.0 g) (1:1 ratio mixture of compound Int-7 and bis(pinacolato)diboran). ESI (m/z) [C.sub.16H.sub.20BClO.sub.4+H].sup.+ 323.

Intermediate 8

Preparation of methyl 4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-carboxylate (Int-8)

##STR00314##

[0510] Preparation of (Int-8): To a solution of compound Int-1 (100 mg, 0.232 mmol) in DMF (2.0 mL) and water (0.5 mL) was added commercial available methyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (60.8 mg, 0.232 mmol) at room temperature. After the solution was degassed with Argon for 10 min, K.sub.3PO.sub.4 (98 mg, 0.464 mmol) and Pd(PPh.sub.3).sub.4 (13.4 mg, 0.012 mmol) were added. After degassing with Argon for 5 min, the reaction mixture was heated to 100 C. and stirred for 16 h. After hot filtration, the filtrate was added water (10 mL), and the precipitated brown solid was collected by filtration. The solid was washed with CH.sub.3CN (2.0 mL) and dried, to afford compound Int-8 (77.0 mg, 80%) as a yellow solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 8.09 (d, J=6.5 Hz, 2H), 7.64 (d, J=6.5 Hz, 2H), 7.54 (d, J=8.5 Hz, 2H), 7.35-7.25 (m, 7H), 5.09 (s, 2H), 4.71 (br s, 1H), 3.93 (s, 3H), 3.24 (t, J=6.5 Hz, 2H), 2.68 (t, J=6.5 Hz, 2H), 1.68-1.56 (m, 4H).

Intermediates 9-13

[0511] Preparation of Intermediates 9-13: In an analogous procedure used for preparation of Int-8, Intermediates 9-13 were synthesized from Int-1 and the corresponding borate esters. For Int-9 the borate ester was commercially available, and Intermediates 3-6 were used for synthesis of Intermediates 10-13:

TABLE-US-00031 Int-# Structure [M + H].sup.+ 9 [00315] [C.sub.27H.sub.29NO.sub.4 + H].sup.+ = 432 10 [00316] [C.sub.28H.sub.31NO.sub.4 + H].sup.+ = 446 11 [00317] [C.sub.30H.sub.35NO.sub.4 + H].sup.+ = 474 12 [00318] [C.sub.28H.sub.31NO.sub.4 + H].sup.+ = 446 13 [00319] [C.sub.29H.sub.33NO.sub.4 + H].sup.+ = 460

Alternative Preparation of Intermediate 11

Preparation of ethyl 4-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)butanoate (Int-11)

##STR00320##

[0512] Preparation of Int-11: A solution of Int-48 (200 mg, 0.489 mmol) and ethyl 4-(4-bromophenyl)butanoate (132 mg, 0.489 mmol) in toluene/EtOH (4 mL/1 mL) was degassed for 10 min under Argon atmosphere followed by the addition of K.sub.2CO.sub.3 (135 mg, 0.977 mmol), and Pd(PPh.sub.3).sub.4 (0.05 equiv) at room temperature. The resulting mixture was degassed with Argon for another 10 min. The resulting mixture was refluxed at 110 C. with stirring for 16 h. After filtration, the solvent was removed. The residue was dissolved in CH.sub.2Cl.sub.2 (5 mL), then hexanes (15 mL) was added, and the solvent volume was reduced to 10 mL by concentration. The solid was filtered out, to afford Int-11 (185 mg, 80%) as a brown solid: ESI (m/z) [C.sub.30H.sub.35NO.sub.4+H].sup.+ 474.

Intermediate 14

Preparation of ethyl 3-(4-(3-(((benzyloxy)carbonyl)amino)propyl)-[1,1-biphenyl]-4-yl)propanoate (Int-14)

##STR00321##

[0513] Preparation of Int-14a: To a solution of 3-(4-bromophenyl)propan-1-amine (800 mg, 3.74 mmol) in DCM (30 mL) was added DIPEA (1.44 g, 11.2 mmol) and CbzCl (637 mg, 3.74 mmol). The reaction mixture was stirred at rt for 2 hr. Water (30 mL) was added, the water layer was extracted with DCM (30 mL2), the combined DCM layers were dried then concentrated, to afford compound Int-14a (1.27 g, 94%) as yellow oil, which was changed to a yellow solid after standing at rt overnight: ESI (m/z) [C.sub.17H.sub.18BrN.sub.2+H].sup.+ 348.

[0514] Preparation of Int-14b: To a solution of ethyl 3-(4-bromophenyl)propanoate (2.00 g, 7.81 mmol) and bis-pinicol borane (2.96 g, 11.6 mmol) in dioxane (20 mL) was added AcOK (2.29 g, 23.3 mmol) and Pd(dppf)Cl.sub.2 (285 mg, 0.389 mmol) at rt. After degassing with Argon for 15 min, the reaction mixture was warmed to 100 C. and stirred for 6 h. After filtration, the filtrate was concentrated and purified by silica column (0% to 50% of EtOAc in DCM, product came out at 10%), to afford compound Int-14b (2.04 g, 86%) as a brown oil: ESI (m/z) [C.sub.17H.sub.25BO.sub.4+H].sup.+ 305.

[0515] Preparation of Int-14: To a solution of Int-14b (750 mg, 2.46 mmol) and Int-14a (859 mmol, 2.46 mmol) in toluene (20 mL) and EtOH (2.5 mL) was added K.sub.2CO.sub.3 (484 mg, 4.93 mmol) and palladium tetrakis (285 mg, 0.247 mmol) at rt. After degassing with Argon for 15 min, the reaction mixture was heated under Argon protection to 110 C. and stirred for 16 h. After filtration, the reaction mixture was partitioned between EtOAc (75 mL) and water (75 mL), the water layer was extracted with EtOAc (75 mL2), the organic layers were combined, dried and concentrated, to afford compound Int-14 (1.02 g, 93%) as brown oil: ESI (m/z) [C.sub.28H.sub.31NO.sub.4+H].sup.+ 446.

Intermediate 15

Preparation of ethyl 3-(4-(5-(((benzyloxy)carbonyl)amino)pent-1-yn-1-yl)-[1,1-biphenyl]-4-yl)propanoate (Int-15)

##STR00322##

[0516] Preparation of Int-15a: To a solution of compound Int-14b (150 mg, 0.493 mmol) and 1-bromo-4-iodobenzene (167 mg, 0.592 mmol) in toluene (4.0 mL) and EtOH (0.5 mL) was added K.sub.2CO.sub.3 (97.0 mg, 0.986 mmol) and palladium tetrakis (57.0 mg, 0.049 mmol) at room temperature. After degassing with Argon for 15 min, the reaction mixture was heated under Argon protection to 110 C. and stirred for 16 h. After filtration, the reaction mixture was partitioned between EtOAc (15 mL) and water (15 mL). The water layer was extracted with EtOAc (15 mL2), and the organic layers were combined, dried, concentrated, and purified by silica column (0% to 60% of EtOAc in DCM, product came out at 20%), to afford Int-15a (144 mg, 88%) as brown oil: ESI (m/z) [C.sub.17H.sub.17BrO.sub.2+H].sup.+ 333.

[0517] Preparation of Int-15: To a solution of Int-15a (100 mg, 0.301 mmol) and benzyl pent-4-yn-1-ylcarbamate (98 mg, 0.450 mmol) in CH.sub.3CN (2.0 mL) was added CuI (2.86 mg, 0.015 mmol), Et.sub.3N (121 mg, 1.20 mmol), tBu.sub.3P (12.1 mg, 0.060 mmol, 1 N toluene solution) and palladium tetrakis (34.7 mg, 0.030 mmol) at room temperature. After degassing with Argon for 15 min, the reaction mixture was refluxed under Argon protection for 16 h. Solvent was removed, and the residue was purified by silica column (0% to 60% of EtOAc in DCM, product came out at 20%), to afford Int-15 (84 mg, 60%) as a [0518] brown solid: ESI (m/z) [C.sub.3H.sub.31NO.sub.4+H].sup.+ 470.

Intermediate 16

Preparation of benzyl (4-(4-(3-hydroxypropyl)-[1,1-biphenyl]-4-yl)butyl)carbamate (Int-16)

##STR00323##

[0519] Preparation of Int-16a: To a stirred solution of commercially available methyl 3-(4-bromophenyl)propanoate (2.50 g, 10.28 mmol) in THF (25 mL), was added LAH (2.5M in THF solution; 0.78 mL, 20.56 mmol) at 0 C. under nitrogen and stirred at ambient temperature for 2 h. The reaction mixture was then quenched with sat. aq NH.sub.4Cl solution (20 mL) and EtOAc (20 mL) at 0 C. The aqueous layer was extracted with EtOAc (350 mL) and combined organic layer was washed with brine (25 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The crude was purified by silica gel combi-flash column chromatography using 20% EtOAc in Hexanes to afford compound Int-16a (1.90 g, 86%) as colorless liquid. .sup.1H NMR (400 MHz, CDCl3) 7.41 (d, J=12.0 Hz, 2H), 7.09 (d, J=12 Hz, 2H), 3.68-3.64 (m, 2H), 2.68 (t, J=8.0 Hz, 2H), 1.91-1.82 (m, 2H).

[0520] Preparation of Int-16b: To a stirred solution of compound Int-16a (1.0 g, 4.62 mmol) in DMF/H.sub.2O (10 mL; 4:1), was added bis-pinacolato-diboron (2.59 g, 6.01 mmol) followed by K.sub.3PO.sub.4 (1.96 g, 9.24 mmol) at ambient temperature under nitrogen. The reaction mixture was stirred for 5 min, degassed and refilled with nitrogen. Then to the reaction mixture was added Pd(PPh.sub.3).sub.4 (0.26 g, 0.23 mmol) under nitrogen and reaction was heated to 90 C. and stirred for 16 h. The reaction mixture was quenched with water (50 mL) and extracted with EtOAc (350 mL). Combined organic layer was washed with brine (25 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The crude was purified by silica gel combiflash column chromatography using 35% EtOAc in Hexanes to afford Int-16b (0.80 g, 66%) as pale-yellow solid. .sup.1H NMR (400 MHz, CDCl3) 7.74 (d, J=8.0 Hz, 2H), 7.20 (d, J=8 Hz, 2H), 3.69-3.65 (m, 2H), 2.69 (t, J=12.0 Hz, 2H), 1.91-1.84 (m, 2H), 1.24 (s, 12H).

[0521] Preparation of Int-16: To a stirred solution of compound Int-16b (1.0 g, 3.81 mmol) in DMF/H.sub.2O (10 mL; 4:1), was added compound Int-1 (2.10 g, 4.96 mmol) followed by K.sub.3PO.sub.4 (1.61 g, 7.63 mmol) at ambient temperature under nitrogen. The reaction mixture was stirred for 5 min, degassed and refilled with nitrogen. Then to the reaction mixture was added Pd(PPh.sub.3).sub.4 (0.22 g, 0.19 mmol) under nitrogen and reaction was stirred at 90 C. for 16 h. The reaction mixture was quenched with water (50 mL) and extracted with EtOAc (350 mL). Combined organic layer was washed with brine (25 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude was purified by silica gel combi-flash column chromatography using 45% EtOAc in hexanes to afford compound Int-16 (0.90 g, 56%) as off-white solid. ESI (m/z) [C.sub.27H.sub.31NO.sub.3+H].sup.+ 418.

Intermediate 17

Preparation of tert-butyl (3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propyl)carbamate (Int-17)

##STR00324##

[0522] Preparation of Int-17a: To a solution of 3-(4-bromophenyl)propan-1-amine (800 mg, 3.74 mmol) in CH.sub.2Cl.sub.2 (30 mL) was added DIPEA (1.44 g, 11.2 mmol) and (Boc).sub.2O (816 mg, 3.74 mmol). The reaction mixture was stirred at rt for 2 h. Water (30 mL) was added, the water layer was extracted with DCM (30 mL2), the combined DCM layers were dried and concentrated, to afford compound Int-17a (1.27 g, 94%) as a yellow solid: ESI (m/z) [C.sub.14H.sub.20BrNO.sub.2+H].sup.+ 314.

[0523] Preparation of Int-17b: To a stirred solution of Int-17a (0.75 g, 2.39 mmol) and bis-pinacolato-diboron (0.73 g, 2.87 mmol) in 1,4-dioxane (10 mL) was added KOAc (0.7 g, 7.18 mmol) at room temperature. The reaction mixture was degassed for 10 min with argon then Pd(dppf).sub.2Cl.sub.2.Math.DCM (89 mg, 0.095 mmol) was added. The reaction mixture was warmed to 100 C. and stirred for 6 h. Reaction mixture was cooled to room temperature and filtered via celite bed, washed with EtOAc and the filtrate was concentrated under reduced pressure. The obtained crude was purified by combiflash (40 g column) chromatography eluting with 10% EtOAc in Hexanes to afford compound Int-17b (0.5 g, 58%) as an off-white solid. ESI (m/z) [C.sub.20H.sub.32BNO.sub.4+H].sup.+ 362.

[0524] Preparation of Int-17c: To a stirred solution of compound Int-17b (0.40 g, 1.1 mmol) and Int-1 (0.60 g, 1.39 mmol) in DMF:H.sub.2O (6 mL:2 mL) was added K.sub.3PO.sub.4 (0.88 g, 4.15 mmol) at room temperature. The reaction mixture was degassed for 10 min with argon. Pd(PPh.sub.3).sub.4 (50.8 mg, 0.044 mmol) was added, then warmed to 100 C. and stirred for 16 h. The reaction mixture was cooled to room temperature, filtered via celite bed, washed with EtOAc, and the filtrate was concentrated under reduced pressure. The obtained crude was purified by combiflash (40 g column) chromatography eluting with 25% EtOAc in Hexanes to afford compound Int-17c (0.50 g, 70%) as an off-white solid. ESI (m/z) [C.sub.32H.sub.40N.sub.2O.sub.4+H].sup.+ 517.

[0525] Preparation of Int-17: To a stirred solution of compound Int-17c (2.0 g, 3.87 mmol) in dichloromethane (20 mL) was added TFA (3 mL, 1.5 wt/v) at 0 C. The reaction mixture was warmed to room temperature and stirred for 2 h. Reaction mixture was concentrated under reduced pressure, and the obtained crude was co-distilled with dichloromethane (10 mL) and MTBE (10 mL), then dried under reduced pressure to afford compound Int-17 (1.80 g, TFA salt) as an off-white solid. ESI (m/z) [C.sub.29H.sub.32F.sub.3N.sub.2O.sub.3+H].sup.+ 417.

Intermediate 18

Preparation of benzyl (4-(4-(3-(2-oxoacetamido)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamate (Int-18)

##STR00325##

[0526] Preparation of Int-18a: To a stirred solution of compound Int-17 (400 mg, 0.974 mmol) in DCM (20 mL) was added DIPEA (254 mg, 1.958 mmol) followed by the addition of acetoxy acetyl chloride (131 mg, 0.9754 mmol) at room temperature. Stirring continued for 2 h. After completion of the reaction, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (250 mL). The organic layer was washed with brine solution, dried over anhydrous Na.sub.2SO.sub.4, filtered, and evaporated under reduced pressure to obtain crude product Int-18a, used as such for next step without purification. ESI (m/z) [C.sub.31H.sub.36N.sub.2O.sub.5+H].sup.+ 517

[0527] Preparation of Int-18b: To a stirred solution of compound Int-18a (500 mg, 0.968 mmol) in MeOH (50 mL) was added K.sub.2CO.sub.3 (270 mg, 1.937 mmol). The reaction mixture was stirred for 2 h at room temperature. After completion of the reaction, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (350 mL). The combined organic layer was washed with brine solution (10 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and evaporated under reduced pressure to obtain crude product which was purified by flash column chromatography (silica gel 120 g column) using 50% EtOAc in hexanes to afford compound Int18b (350 mg, 70%) as an off-white solid. ESI (m/z) [C.sub.29H.sub.34N.sub.2O.sub.4+H].sup.+ 475.

[0528] Preparation of Int-18: To a stirred solution of compound Int-18b (350 mg, 0.73 mmol) in dichloromethane (15 mL) was added portion wise Dess-Martin periodinane (470 mg, 1.1 mmol) at 0 C. The reaction mixture was warmed to ambient temperature and stirring continued for 2 h. Then reaction mixture was quenched with aqueous sodium thiosulfate solution (sat. solution, 10 mL) and the separated solid was filtered through celite pad which was washed with dichloromethane (50 mL). The filtrate was concentrated to afford compound Int-18 (200 mg, crude) and used as such for next step without purification. ESI (m/z) [C.sub.29H.sub.32N.sub.2O.sub.4+H].sup.+ 473

Intermediate 19

Preparation of ethyl 2-((4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)oxy)acetate (Int-19)

##STR00326##

[0529] Preparation of Int-19a: To a stirred solution of 4-hydroxy-phenylboronic acid (3.0 g, 21.7 mmol) and Int-1 (14.05 g, 32.6 mmol) in a mixture of DMF/H.sub.2O (90:30 mL) was added to K.sub.3PO.sub.4 (9.20 g, 43.4 mmol) at room temperature. The reaction mixture was degassed for 10 min with argon. Pd(PPh.sub.3).sub.4 (0.50 g, 0.43 mmol) was added. The reaction mixture was warmed to 100 C. and stirred for 16 h. The reaction mixture was cooled to room temperature, filtered through celite bed and washed with EtOAc (100 mL). The filtrate was concentrated under reduced pressure and the residue was purified by combi flash column (220 g column size) chromatography eluting with 25% EtOAc in hexanes to afford Int-19a (5.20 g, 63%) as an off-white solid. ESI (m/z) [C.sub.24H.sub.25NO.sub.3+H].sup.+ 376.

[0530] Preparation of Int-19: To a stirred solution of compound Int-19a (4.50 g, 12.0 mmol) and ethyl bromoacetate (2.30 g, 14.4 mmol) in acetone (60 mL) was added to K.sub.2CO.sub.3 (2.50 g, 18 mmol) at 0 C. The reaction mixture was warmed to ambient temperature and stirred for 16 h. The reaction mixture was filtered through celite, washed with EtOAc (200 mL) and the filtrate evaporated under reduced pressure to obtain the crude product. The crude was purified by flash column chromatography (silica gel 120 g column) using 15-20% EtOAc in hexanes to afford compound Int-19 (4.20 g, 78%) as an off-white solid. ESI (m/z) [C.sub.28H.sub.31NO.sub.5+H].sup.+ 462.

Intermediate 20

Preparation of (2R,2R,3R,3R,4R,4R,5S,5'S)-6,6-((2-aminoethyl)azanediyl)bis(hexane-1,2,3,4,5-pentaol) (Int-20)

##STR00327##

[0531] Preparation of Int-20a: To a solution of commercially available benzyl (2-aminoethyl) carbamate (500 mg, 2.57 mmol) in MeOH (10 mL) was added D-glucose (1.39 g, 7.72 mmol), AcOH (464 mg, 7.72 mmol) and NaCNBH.sub.3 (485 mg, 7.72 mmol). The resulting solution was stirred at 60 C. for 3 h. Additional D-glucose (926 mg, 5.14 mmol), AcOH (308 mg, 5.14 mmol) and NaCNBH.sub.3 (322 mg, 5.14 mmol) were added. The reaction mixture was heated at 60 C. for 2 h. Solvent was removed and the residue was dissolved in water (3.0 mL). Saturated Na.sub.2CO.sub.3 (1.0 mL) was added, then the pH of solution was adjusted to 3 using 4 N HCl. The solution was purified by reverse phase column (5%90% of CH.sub.3CN in 0.05 M HCl, product came out at 10%30%), to afford Int-20a (896 mg, 62%) as colorless syrup. .sup.1H NMR (500 MHz, CD.sub.3OD) 7.35-7.34 (m, 5H), 5.11 (s, 2H), 4.20 (br s, 2H), 3.83-3.29 (m, 18H).

[0532] Preparation of Int-20: Compound Int-20a (896 mg, 1.63 mmol) was dissolved in MeOH (20 mL). After purging with nitrogen, 10% Pd/C (300 mg) was added. The suspension was stirred under hydrogen (balloon) for 6 h. Pd/C was filtered off and the solids were washed with MeOH/water (1:1, 10 mL). The combined filtrate was concentrated to afford Int-20 (633 mg, 93%) as colorless syrup. .sup.1H NMR (500 MHz, CD.sub.3OD) 4.29-4.28 (m, 2H), 3.90-3.67 (m, 12H), 3.56-3.50 (m, 6H).

Intermediate 21

Preparation of (2R,2R,3R,3R,4R,4R,5S,5'S)-6,6-((3-aminopropyl)azanediyl)bis(hexane-1,2,3,4,5-pentaol) (Int-21)

##STR00328##

[0533] Preparation of Int-21: Int-21 was prepared in a procedure analogous to Int-20 substituting benzyl (3-aminopropyl)carbamate as the starting material: .sup.1H NMR (500 MHz, D.sub.2O) 3.99 (br s, 2H), 3.80-3.60 (m, 10H), 3.09-2.84 (m, 8H), 1.96-1.87 (m, 2H); ESI (m/z) [C.sub.15H.sub.34N.sub.2O.sub.10+H].sup.+ 403.

Intermediate 22

Preparation of (2R,2R,3R,3R,4R,4R,5S,5'S)-6,6-((4-aminobutyl)azanediyl)bis(hexane-1,2,3,4,5-pentaol) (Int-22)

##STR00329##

[0534] Preparation of Int-22: Int-22 was prepared in a procedure analogous to Int-20 substituting benzyl (4-aminobutyl)carbamate as the starting material: .sup.1H NMR (500 MHz, D.sub.2O) 4.03-4.00 (m, 2H), 3.79-3.54 (m, 10H), 3.00-2.95 (m, 8H), 1.69-1.65 (m, 4H); ESI (m/z) [C.sub.16H.sub.36N.sub.2O.sub.10+H].sup.+ 417.

Intermediate 23

Preparation of 3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanoic acid (Int-23)

##STR00330##

[0535] Preparation of Int-23: To a stirred solution of Int-10 (30.0 g, 67.4 mmol) in a mixture of THF/MeOH (300 mL:60 mL) was added a solution of LiOH.Math.H.sub.2O (5.60 g, 134.8 mmol) in H.sub.2O (60 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 h. Solid formation was observed. The reaction mixture concentrated under reduced pressure. The obtained solid was taken in water (500 mL), acidified to pH=2 with 2N HCl solution and stirred for 30 min and the solids were filtered, washed with hexanes (200 mL) and dried under reduced pressure to afford Int-23 (26 g, 89%) as an off-white solid. ESI (m/z) [C.sub.27H.sub.29NO.sub.4+H].sup.+ 432.

Intermediate 24

Preparation of methyl (S)-2-amino-4-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) amino)butanoate (Int-24)

##STR00331##

[0536] Preparation of Int-24a: To a solution of commercially available methyl (S)-4-amino-2-(((benzyloxy)carbonyl)amino)butanoate (200 mg, 0.661 mmol) in MeOH (2.0 mL) was added D-glucose (595 mg, 3.30 mmol) followed by AcOH (278 mg, 4.62 mmol) and NaCNBH.sub.3 (208 mg, 3.30 mmol). The resulting solution was stirred at 50 C. for 6 h. Additional D-glucose (119 mg, 0.661 mmol), AcOH (79.3 mg, 1.32 mmol) and NaCNBH.sub.3 (42.3 mg, 0.661 mmol) were added, and the reaction mixture was heated to 50 C. and stirred for 16 h. Desired product Int-24a was formed along with mono-polyol self-cyclized by-product. The solvent was removed, the residue was azeotroped with 2 N HCl in MeOH (10 mL) twice, and then was purified by reverse phase column (5% to 80% of CH.sub.3CN in H.sub.2O, product came out at 20%), to afford compound Int-24a (256 mg, 61%) as colourless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.35-7.34 (m, 5H), 5.10-5.09 (m, 2H), 4.35-4.30 (m, 1H), 3.95 (br s, 2H), 3.78-3.60 (m, 12H), 3.50-3.40 (m, 5H), 2.90-2.60 (m, 4H), 2.20-1.90 (m, 2H).

[0537] Preparation of Int-24: To a solution of compound Int-24a (200 mg, 0.317 mmol) in MeOH (4.0 mL) was added 20% Pd(OH).sub.2 (60 mg). The resultant mixture was stirred under hydrogen (balloon) at room temperature for 16 h. After filtration, the solvent was removed, to afford compound Int-24 (141 mg, 90%) as colorless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 4.25-4.15 (m, 1H), 4.02-3.95 (m, 2H), 3.85 (s, 3H), 3.78-3.62 (m, 10H), 3.05 (s, 1H), 2.90-2.80 (m, 3H), 2.70-2.55 (m, 2H), 2.30-2.20 (m, 1H), 2.05-1.95 (m, 1H).

Intermediate 25

Preparation of methyl (S)-2-amino-4-(dimethylamino)butanoate (Int-25)

##STR00332##

[0538] Preparation of Int-25a: To a solution of (S)-4-amino-2-((tert-butoxycarbonyl)amino)butanoic acid (500 mg, 2.29 mmol) in MeOH (10 mL) was added formaldehyde (37% in H.sub.2O, 275 mg, 9.16 mmol) followed by AcOH (550 mg, 9.16 mmol) and NaCNBH.sub.3 (576 mg, 9.16 mmol). The resulting solution was stirred at room temperature for 4 h. After solvent was removed, the residue was re-dissolved in H.sub.2O (10 mL), and 1N NaOH (2.0 mL) was added (pH around 12). Then pH value of the solution was slowly adjusted to 5 using 1 N HCl. The solution was purified by reverse phase column (5% to 90% of CH.sub.3CN in H.sub.2O, product came out at 40%), to afford compound Int-25a (340 mg, 60%) as a colourless sticky solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 4.07-4.06 (m, 1H), 3.34-3.14 (m, 2H), 2.87 (s, 6H), 2.22-2.19 (m, 1H), 2.03-1.92 (m, 1H), 1.45 (s, 9H).

[0539] Preparation of Int-25: To a solution of compound Int-25a (338 mg, 1.37 mmol) in MeOH (6.0 mL) was added SO.sub.2C.sub.1 (1.0 mL). The resultant mixture was refluxed at 70 C. for 6 h. The solvent was removed, to afford compound Int-25 (315 mg, 98%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 4.26-4.23 (m, 1H), 3.90 (s, 3H), 3.44-3.32 (m, 2H), 2.94 (s, 6H), 2.43-2.29 (m, 2H).

Intermediate 26

Preparation of benzyl (4-(4-(3-aminopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamate (Int-26)

##STR00333##

[0540] Preparation of Int-26: To a solution of (S)-4-amino-2-((tert-butoxycarbonyl)amino)butanoic acid (800 mg, 3.67 mmol) in THF (15 mL), MeOH (15 mL) and water (5.0 mL) was added NaHCO.sub.3 (616 mg, 7.33 mmol) followed by FmocCl (948 mg, 3.67 mmol) at 0 C. The resulting solution was stirred at 0 C. for 1 h, and then at room temperature for 16 h. After the solvent was removed, the residue was washed with hexanes, partitioned with EtOAc (30 mL) and H.sub.2O (30 mL). The water layer was extracted with EtOAc (30 mL2). The organic layers were combined and concentrated, to afford compound Int-26 (1.54 g, 95%) as a white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.79-7.78 (m, 2H), 7.65-7.63 (m, 2H), 7.39-7.28 (m, 4H), 4.34-4.33 (m, 2H), 4.22-4.20 (m, 1H), 3.50-3.40 (m, 1H), 3.31-3.16 (m, 2H), 2.10 (br s, 1H), 1.85-1.75 (m, 1H), 1.44 (s, 9H); ESI (m/z) [C.sub.24H.sub.28N.sub.2O.sub.6+H].sup.+ 441.

Intermediate 27

[0541] Preparation of tert-butyl ((S)-1-((3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propyl)amino)-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)-1-oxohexan-2-yl)carbamate (Int-27)

##STR00334##

[0542] Preparation of Int-27a: To a solution of N6-(((9H-fluoren-9-yl)methoxy)carbonyl)-N2-(tert-butoxycarbonyl)-L-lysine (646 mg, 1.38 mmol) in DMF (20 mL) was added HATU (577 mg, 1.51 mmol), compound Int-17 (625 mg, 1.38 mmol) and DIPEA (535 mg, 4.14 mmol) at room temperature. The resultant mixture was stirred at room temperature for 1 h. Water (150 mL) was added, and the water layer was extracted with EtOAc (150 mL3). The organic layers were combined and concentrated. Water (20 mL) was added to the residue, and the resulting precipitate was collected by filtration and dried, to afford compound Int-27a (1.09 g, 91%) as a white solid: .sup.1H NMR (300 MHz, CDCl.sub.3) 7.78-7.72 (m, 2H), 7.57-7.52 (m, 2H), 7.50-7.20 (m, 17H), 6.12 (br s, 1H), 5.09 (s, 2H), 4.82 (br s, 1H), 4.72 (br s, 1H), 4.41-4.38 (m, 2H), 4.20-4.13 (m, 1H), 3.98 (br s, 1H), 3.31-3.17 (m, 6H), 2.69-2.63 (m, 4H), 1.89-1.43 (m, 7H), 1.42 (s, 9H), 1.38-1.25 (m, 5H); ESI (m/z) [C.sub.38H.sub.62N.sub.4O.sub.7+H].sup.+ 867.

[0543] Preparation of Int-27b: To a solution of compound Int-27a (1.08 g, 1.24 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added piperidine (4.0 mL). The resulting mixture was stirred at room temperature for 3 h. After the solvent was removed, MTBE/hexanes (20 mL/20 mL) was added. The resulting solid was collected and washed with hexanes (20 mL), to afford compound Int-27b (765 mg, 95%) as a white solid: .sup.1H NMR (300 MHz, CDCl.sub.3) 7.50-7.47 (m, 4H), 7.34-7.32 (m, 5H), 7.22-7.20 (m, 4H), 6.60 (br s, 1H), 5.20 (br s, 1H), 5.14 (s, 2H), 4.76 (br s, 1H), 4.10 (br s, 2H), 3.31-3.22 (m, 4H), 2.80-2.55 (m, 6H), 1.89-1.43 (m, 12H), 1.43 (s, 9H); ESI (m/z) [C.sub.38H.sub.52N.sub.4O.sub.5+H].sup.+ 645.

[0544] Preparation of Int-27: To a solution of compound Int-27b (100 mg, 0.155 mmol) in MeOH (3.0 mL) was added D-glucose (84.0 mg, 0.465 mmol) followed by AcOH (27.9 mg, 0.465 mmol) and NaCNBH.sub.3 (29.2 mg, 0.465 mmol). The resulting solution was stirred at 50 C. for 16 h. Additional D-glucose (55.8 mg, 0.310 mmol), AcOH (18.6 mg, 0.310 mmol) and NaCNBH.sub.3 (19.8 mg, 0.310 mmol) were added, and the reaction mixture was heated at 50 C. and stirred for 8 h. After the solvent was removed, water was added to the residue. The resulting solid was collected by filtration and dried, to afford compound Int-27 (130 mg, 86%) as an off-white solid, which was a boran complex: ESI (m/z) [C.sub.50H.sub.76N.sub.4O.sub.15+H].sup.+ 973.

Intermediate 28

Preparation of tert-butyl ((S)-1-((3-(4-(4-aminobutyl)-[1,1-biphenyl]-4-yl)propyl)amino)-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)-1-oxohexan-2-yl)carbamate (Int-28)

##STR00335##

[0545] Preparation of Int-28: To a solution of compound Int-27 (500 mg, 0.514 mmol) in EtOH (10 mL) and water (5.0 mL) was added 20% Pd(OH).sub.2 (50 mg) and AcOH (185 mg, 3.08 mmol). The resultant mixture was stirred under hydrogen (balloon) at room temperature at 45 C. for 4 h. After filtration, the solvent was removed, to afford compound Int-28 (443 mg, 90%) as a white syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.27-7.26 (m, 4H), 3.98 (br s, 3H), 3.85-3.44 (m, 16H), 2.94-2.91 (m, 4H), 2.73-2.54 (m, 4H), 1.95 (s, 6H), 1.88-1.45 (m, 12H), 1.43 (s, 9H); ESI (m/z) [C.sub.42H.sub.70N.sub.4O.sub.13+H].sup.+ 839.

Intermediate 29

Preparation of benzyl (4-(4-(3-((S)-2-amino-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxy hexyl)amino)hexanamido)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamate (Int-29)

##STR00336##

[0546] Preparation of Int-29: To a solution of compound Int-27 (borane complex, 425 mg, 0.437 mmol) in EtOH (5.0 mL) was added 4 N HCl (10 mL) at room temperature. The resultant mixture was stirred at 45 C. for 2 h. After the solvent was removed, the residue was azeotroped with 1 N HCl in MeOH (5.0 mL) and MeOH (25.0 mL) to remove methylborate, affording compound Int-29 (402 mg, 97%) as a white solid: .sup.1H NMR (300 MHz, CD.sub.3OD) 7.53-7.47 (m, 4H), 7.31-7.21 (m, 9H), 5.05 (m, 2H), 4.16 (br s, 2H), 3.86-3.67 (m, 15H), 3.41-3.11 (m, 6H), 2.73-2.65 (m, 4H), 1.89-1.52 (m, 12H); ESI (m/z) [C.sub.45H.sub.68N.sub.4O.sub.13+H].sup.+ 873.

Intermediate 30

Preparation of (2S,3S,4R,5R,6R)-2-(methoxycarbonyl)-6-(2-oxoethyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (Int-30)

##STR00337##

[0547] Preparation of Int-30a: To a stirred solution of (2R,3S,4R,5R,6R)-2-allyl-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (see Zou, W.; Vembaiyan, K; JOC 78(6), 2703-9, 2013, 10.0 g, 30.3 mmol) in acetonitrile:water (80 mL:80 mL) was added (diacetoxyiodo)benzene (29.27 g, 90.9 mmol) at 0 C. After 10 min, TEMPO (1.40 g, 9.06 mmol) was added, warmed to room temperature, and stirred for 16 h. Reaction was not completed as monitored by TLC. Again added (diacetoxyiodo)benzene (9.7 g, 30.3 mmol) and TEMPO (0.46 g, 0.1 mmol) was added slowly, the reaction mixture was warmed to room temperature and stirred for 16 h. Reaction mixture was quenched with hypo solution (10% aqueous Sodium thiosulfate (Na.sub.2S.sub.2O.sub.3)) at 0 C. and extracted with EtOAc (3500 mL). Combined organic layer was washed with brine (25 mL) and dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-30a (12.0 g, crude) as a brown liquid. ESI (m/z) [C.sub.15H.sub.20O.sub.9+H].sup.+ 345

[0548] Preparation of Int-30b: To a stirred solution of Int-30a (12.0 g, 34.9 mmol) in DMF (150 mL) was added NaHCO.sub.3 (5.80 g, 69.7 mmol) at 0 C. under nitrogen atmosphere. After 10 min, methyl iodide (20.09 g, 139.5 mmol) was added slowly, the reaction mixture was warmed to room temperature and stirred for 16 h. Reaction mixture was quenched with water (500 mL) and extracted with EtOAc (3500 mL). Combined organic layer was washed with brine (50 mL) and dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The obtained crude was purified by combiflash (220 g column) chromatography eluting with 25% EtOAc in hexanes to afford Int-30b (2.4 g, 20%) as an off-white solid. (4.0 g of Int-30a recovered); ESI (m/z) [C.sub.16H.sub.22O.sub.9+H].sup.+ 359.

[0549] Preparation of Int-30: To a cooled solution of compound Int-30b (1.30 g, 3.63 mmol) in a mixture of 1,4-dioxane/water (12 mL, 3:1 ratio) was added 2,6-lutidine (776 mg, 7.26 mmol) followed by NaIO.sub.4 (3.10 g, 14.52 mmol) and then added OSO.sub.4 (19 mg, 0.07 mmol) at 0 C. The resulted mixture was stirred at ambient temperature for 4 h. Reaction mixture was diluted with water (50.0 mL) and EtOAc (50.0 mL), the EtOAc layer was separated and washed with 5% aqueous citric acid solution twice (225.0 mL) followed by brine solution (50.0 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered, and evaporated under reduced pressure to get compound -Int-30 (900 mg, 69%) as a light grey, sticky liquid. ESI (m/z) [C.sub.15H.sub.20NO.sub.10+H].sup.+ 361.

Intermediate 31

Preparation of 3,5-bis(((4R,4R,4S,5R,5'S)-2,2,2,2,2,2-hexamethyl-[4,4:5,4-ter(1,3-dioxolan)]-5-yl)methoxy)aniline (Int-31)

##STR00338##

[0550] Preparation of Int-31a: To a stirred solution of sodium gluconate (10 g, 40.32 mmol) in 2,2-dimethoxy propane (50 mL) was added 4M HCl in MeOH (20 mL) slowly at ambient temperature. The reaction mixture was stirred at ambient temperature for 12 h. The volatiles were removed under reduced pressure and the obtained crude was basified with saturated aqueous NaHCO.sub.3 solution (100 mL) and extracted with MTBE (2100 mL). The combined organic layer was washed with water (50 mL) followed by brine (50 mL). Organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and the volatiles were removed under reduced pressure. The obtained crude was purified by comb-flash column chromatography (silica gel 120 g column), eluted with 10-15% EtOAc in hexanes to afford compound Int-31a (4.0 g, 34%) as a pale-yellow liquid ESI (m/z) [C.sub.17H.sub.28O.sub.8+H].sup.+ 361.

[0551] Preparation of Int-31b: To a stirred solution of compound Int-31a (3.0 g, 8.31 mmol) in dry THF (50 mL) was added 1.7M DIBAL in toluene (15 mL, 24.9 mmol) slowly at 78 C. The reaction mixture was warmed to ambient temperature slowly and stirred for 12 h. The reaction mixture was quenched with water (20 mL) at 0 C. The resulting solid was filtered through a celite pad and washed with EtOAc (250 mL), filtrate was dried over Na.sub.2SO.sub.4, filtered and the volatiles were removed under reduced pressure to obtain the desired compound Int-31b (1.80 g, 60%) as a pale-yellow liquid. ESI (m/z) [C.sub.16H.sub.28O.sub.7+H].sup.+ 333.

[0552] Preparation of Int-31c: To a stirred solution of 1,3-difluoro-5-nitrobenzene (2.0 g, 12.57 mmol) in DMF (20 mL) was added Cs.sub.2CO.sub.3 (16.0 g, 50.31 mmol) slowly at 0 C. followed by the addition of Int-31b (13.0 g, 37.71 mmol). The resulted reaction mixture was heated to 120 C. and stirred for 24 h. The reaction mixture was quenched with ice cold water (50 mL) and extracted with EtOAc (220 mL). The combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and the volatiles were removed under reduced pressure. The obtained crude was purified by flash column chromatography (silica gel 120 g column), eluting with 20-25% EtOAc in hexanes to afford Int-31c (4.50 g, 39%) as a pale-yellow liquid. ESI (m/z) [C.sub.38H.sub.57NO.sub.16+H].sup.+ 784.

[0553] Preparation of Int-31: To a stirred solution of compound Int-31c (4.50 g, 5.71 mmol) in EtOH (100 mL) was added 10% Pd/C (1.80 g, 50% weight substrate) slowly under nitrogen atmosphere. The reaction mixture was warmed to ambient temperature and stirred under H.sub.2 bladder for 12 h. The reaction mixture was filtered through celite pad and washed with EtOAc (2100 mL). The filtrate was concentrated under reduced pressure to obtain the desired compound Int-31 (4.0 g, 87%) as a pale-yellow solid. ESI (m/z) [C.sub.38H.sub.59NO.sub.14+H].sup.+ 754.

Intermediate 32

Preparation of (3,5-bis(((4R,4R,4S,5R,5'S)-2,2,2,2,2,2-hexamethyl-[4,4:5,4-ter(1,3-dioxolan)]-5-yl)methoxy)phenyl)methanamine (Int-32)

##STR00339##

[0554] Preparation of Int-32a: To a stirred solution of NaH 60% in oil (3.2 g, 134 mmol) in DMF (50 mL) at 0 C., was added a solution of 3,5-difluorobenzonitrile (15.0 g, 44.9 mmol) in DMF (20 mL) slowly dropwise at 0 C. The reaction mixture was stirred for 15 minutes. Int-31b (2.5 g, 17.9 mmol) was added then the resultant reaction mixture was stirred in sealed tube at 90 C. for 15 h. The reaction mixture was quenched with saturated NH.sub.4Cl solution and extracted with MTBE (2100 mL). The combined organic layer washed with brine (100 mL), dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure. The obtained crude compound was purified by combi-flash chromatography eluting with 10-100% EtOAc in hexanes (compound isolated at 15% of EtOAc) to afford Int-32a (10.0 g, 73%) as a gummy liquid; ESI (m/z) [C.sub.39H.sub.57NO.sub.14+H].sup.+ 764.

[0555] Preparation of Int-32b: To a stirred solution of compound Int-32a (230 mg, 0.301 mmol) in dry THF (5 mL) was added 2.0 M LAH in THF (6.50 mL, 12.97 mmol) slowly dropwise at 0 C. The reaction mixture was warmed to ambient temperature and stirring was continued for 16 h under N.sub.2 atmosphere. Reaction mixture was cooled to 0 C., quenched with aqueous NH.sub.4Cl solution and filtered through celite pad. The filtrate was diluted with EtOAc (30 mL) and water (30 mL), separated the EtOAc layer, the aqueous layer again washed with EtOAc (250 mL), then the combined organic layer was washed with brine solution (50 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered, and evaporated under reduced pressure yielding crude which was purified by flash chromatography (0-5% MeOH in dichloromethane) to afford Int-32b (160 mg, 69%) as an off-white sticky solid. ESI (m/z) [C.sub.39H.sub.61NO.sub.14+H].sup.+ 768.

[0556] Preparation of Int-32: To a stirred solution of compound Int-32b (160 mg, 0.208 mmol) in MeOH (5 mL) was added 4M HCl in methanol (5.0 mL) at 0 C. The reaction mixture was stirred at room temperature for 12 h. The reaction mixture was concentrated under reduced pressure and the obtained crude was purified by reverse phase comb-flash chromatography eluting with (20-40% CH.sub.3CN in H.sub.2O with 0.1% HCl buffer) to afford Int-32 (104 mg, 88%) as an off-white solid. .sup.1H NMR (400 MHz, D.sub.2O) 6.64 (s, 3H), 4.20-4.18 (m, 2H), 4.12-4.08 (m, 2H), 4.04-3.98 (m, 6H), 3.85-3.82 (m, 2H), 3.74-3.70 (m, 6H), 3.60-3.55 (m, 2H); ESI (m/z) [C.sub.21H.sub.37NO.sub.14+H].sup.+ 528; HPLC purity 93% (method C at 230 nm).

Intermediate 33

Preparation of 3-(3,5-bis(((4R,4R,4S,5R,5'S)-2,2,2,2,2,2-hexamethyl-[4,4:5,4-ter(1,3-dioxolan)]-5-yl)methoxy)phenyl)propan-1-amine (Int-33)

##STR00340##

[0557] Preparation of Int-33a: To a stirred solution of Int-32a (10.0 g, 13.1 mmol) in CH.sub.2Cl.sub.2 (100 mL), was charged 1M DIBAL in toluene (26.2 mL, 26.2 mmol) dropwise at 78 C. The reaction mixture was stirred at 78 C. for 30 minutes. The reaction mixture was quenched with saturated NH.sub.4Cl solution, diluted with CH.sub.2Cl.sub.2 (100 mL) and filtered through celite bed. The filtrate was washed with brine (100 mL), dried over Na.sub.2SO.sub.4 filtered, and concentrated under reduced pressure. Obtained crude compound was purified by combi-flash chromatography eluting with 10-100% EtOAc in hexanes (compound isolated at 20% of EtOAc) to afford compound Int-33a (7.0 g, 70%) as a gummy oil; ESI (m/z) [C.sub.39H.sub.58O.sub.15+H].sup.+ 767.

[0558] Preparation of Int-33b: A stirred solution of Int-33a (7.0 g, 9.14 mmol) in toluene (70 mL), was charged PPh.sub.3CHCOOCH.sub.3 (4.60 g, 13.7 mmol) at room temperature. The resultant reaction mixture was warmed to 60 C. and stirred for 15 h. The reaction mixture was concentrated, the obtained crude stirred in MTBE (100 mL) and filtered to remove Ph.sub.3PO. The filtrate was concentrated and crude material was purified by combi-flash chromatography eluting with (30% EtOAc in hexanes) to afford Int-33b (7.20 g, 96%) as a sticky oil; ESI (m/z) [C.sub.42H.sub.62O.sub.16+H].sup.+ 823.

[0559] Preparation of Int-33c: A stirred solution of Int-33b (7.20 g, 8.75 mmol) in IPA (70 mL) was charged 10% Pd on carbon 50% wet (1.40 g, 50% weight substrate). The reaction mixture was stirred under H.sub.2 (balloon) pressure for 5 h. The reaction mixture was filtered through celite bed and the filtrate was concentrated under reduced pressure to afford Int-33c (7.0 g, 97%) as a gummy liquid; ESI (m/z) [C.sub.42H.sub.64O.sub.16+H].sup.+ 825.

[0560] Preparation of Int-33d: To a stirred solution of Int-33c (7.0 g, 8.5 mmol) in MeOH (35 mL) and H.sub.2O (35 mL), was charged with LiOH.Math.H.sub.2O (535 mg, 12.7 mmol). The reaction mixture was stirred at room temperature for 15 h. The reaction mixture was concentrated under reduced pressure, the obtained crude diluted with water (100 mL), acidified to pH=3 with 2N HCl solution and extracted with EtOAc (2100 mL). Combined organic layer was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-33d (6.70 g, 98%) as a gummy liquid; ESI (m/z) [C.sub.41H.sub.62O.sub.16+H].sup.+ 810.

[0561] Preparation of Int-33e: To a stirred solution of Int-33d (3.0 g, 3.7 mmol) in THF (30 mL) at 0 C., was added Et.sub.3N (1.5 mL, 11.1 mmol) followed by ethyl chloroformate (0.53 mL, 5.55 mmol). The reaction mixture was stirred for 30 minutes then NH.sub.4OH (15 mL) was added and stirring continued at room temperature for 1 h. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (250 mL). The combined organic layer was washed with brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-33e (3.00 g (crude)) as a gummy liquid; [0562] ESI (m/z) [C.sub.41H.sub.63NO.sub.15+H].sup.+ 810.

[0563] Preparation of Int-33: To a stirred solution of Int-33e (2.50 g, 3.09 mmol) in THF (25 mL), was charged with 2M LAH in THF (2.3 mL, 4.63 mmol) at 0 C. The reaction was warmed to room temperature and stirred for 15 h. The reaction mixture was quenched with saturated NH.sub.4Cl solution (50 mL) and extracted with EtOAc (250 mL). Combined organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-33 (2.30 g, 95%) as a gummy liquid; ESI (m/z) [C.sub.41H.sub.65NO.sub.14+H].sup.+ 796.

Intermediate 34

Preparation of benzyl (4-(4-(3-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamate (Int-34)

##STR00341##

[0564] Preparation of Int-34a: To a solution of Int-23 (1.00 g, 2.31 mmol), tert-butyl (2-aminoethyl)carbamate (479 mg, 2.433 mmol) and DIPEA (899 mg, 6.95 mmol) in DMF (40 mL) was added HATU (925 mg, 2.43 mmol). The reaction mixture was stirred at room temperature for 2 h. After the solvent was removed, water was added. The white precipitate was filtered and dried, to afford compound Int-34a (1.22 g, 92%) as a white solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 7.49-7.47 (m, 4H), 7.35-7.20 (m, 9H), 6.01 (br s, 1H), 5.09 (s, 2H), 4.71 (br s, 2H), 3.33-3.20 (m, 6H), 2.99 (t, J=7.5 Hz, 2H), 2.66 (t, J=7.5 Hz, 2H), 2.50 (t, J=7.5 Hz, 2H), 1.67-1.58 (m, 4H), 1.41 (s, 9H); ESI (m/z) [C.sub.34H.sub.43N.sub.3O.sub.5+H].sup.+ 574.

[0565] Preparation of Int-34: To a solution of Int-34a (1.12 g, 1.95 mmol) in DCM (30 mL) was added TFA (10 mL). The resultant mixture was stirred at room temperature for 2 h. After the solvent was removed, the residue was granulated with MTBE/hexanes and collected by filtration, to afford Int-34 (1.16 g, 93%) as a yellow solid: ESI (m/z) [C.sub.29H.sub.35N.sub.3O.sub.3+H].sup.+ 474.

Intermediate 35

Preparation of methyl (3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysinate (Int-35)

##STR00342##

[0566] Preparation of Int-35a: To a solution of acid Int-23 (50 mg, 0.116 mmol), methyl N-6-(tert-butoxycarbonyl)-L-lysinate (37.8 mg, 0.127 mmol) and DIPEA (44.9 mg, 0.348 mmol) in DMF (2.0 mL) was added HATU (48.5 mg, 0.127 mmol). The reaction mixture was stirred at room temperature for 2 h. After the solvent was removed, water was added. The white precipitate was filtered and dried, to afford compound Int-35a (69 mg, 88%) as a white solid: ESI (m/z) [C.sub.34H.sub.43N.sub.3O.sub.5+H].sup.+ 573.

[0567] Preparation of Int-35: To a solution of compound Exp-35a (585 mg, 0.868 mmol) in THF (8.0 mL) was added 4 N HCl in dioxane (8.0 mL) at room temperature. The resultant mixture was stirred at room temperature for 3 h. The solvent was removed to afford compound Int-35 (530 mg, 100%) as a white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.50-7.47 (m, 4H), 7.33-7.22 (m, 9H), 5.05 (s, 2H), 4.42-4.40 (m, 1H), 3.68 (s, 3H), 3.14 (t, J=7.0 Hz, 2H), 2.95 (t, J=7.0 Hz, 2H), 2.83 (br s, 2H), 2.66-2.57 (m, 4H), 1.82-1.80 (m, 1H), 1.67-1.26 (m, 9H); ESI (m/z) [C.sub.39H.sub.51N.sub.3O.sub.7+H].sup.+ 574.

Intermediate 36

Preparation of methyl N2-(3-(4-(4-aminobutyl)-[1,1-biphenyl]-4-yl)propanoyl)-N6-(tert-butoxycarbonyl)-L-lysinate (Int-36)

##STR00343##

[0568] Preparation of Int-36: To a cloudy solution of Int-35a (125 mg, 0.186 mmol) in EtOH (2.0 mL) and IPA (2.0 mL) was added 20% Pd(OH).sub.2 (40 mg). The resultant mixture was stirred under hydrogen (balloon) at 40 C. for 4 h. After filtration, the solvent was removed, to afford compound Int-36 (90 mg, 90%) as colorless oil: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.51 (d, J=8.0 Hz, 4H), 7.28-7.24 (m, 4H), 4.36-4.33 (m, 1H), 3.67 (s, 3H), 3.50-3.40 (m, 2H), 2.95 (t, J=7.5 Hz, 2H), 2.76 (t, J=7.5 Hz, 2H), 2.68 (t, J=7.5 Hz, 2H), 2.57 (t, J=7.5 Hz, 2H), 1.72-1.58 (m, 6H), 1.41 (s, 9H), 1.39-1.26 (m, 4H); ESI (m/z) [C.sub.31H.sub.45N.sub.3O.sub.5+H].sup.+ 539.

Intermediate 37

Preparation of methyl N2-(3-(4-(4-aminobutyl)-[1,1-biphenyl]-4-yl)propanoyl)-N6-(tert-butoxycarbonyl)-L-lysinate (Int-37)

##STR00344##

[0569] Preparation of Int-37: To a stirred solution of compound N-Boc-Lys(Boc)OH (570 mg, 1.64 mmol) in CH.sub.2Cl.sub.2 (50.0 mL) was added TEA (250 mg, 2.46 mmol) followed by 1-propanephosphonic anhydride, 50% solution in EtOAc (T.sub.3P, 1.55 g, 4.9 mmol). The reaction mixture was stirred for 15 mins, Int-35 (1.0 g, 1.64 mmol) was added, and the reaction mixture stirred at rt for 6 h. The reaction mixture was concentrated under reduced pressure and the obtained crude material was purified by normal phase column chromatography using 10% MeOH in CH.sub.2Cl.sub.2 to afford compound Int-37 (1.10 g 74%) as off-white solid. ESI (m/z) [C.sub.50H.sub.71N.sub.5O.sub.10+H].sup.+ 902.

Intermediate 38

Preparation of methyl N2-(3-(4-(4-aminobutyl)-[1,1-biphenyl]-4-yl)propanoyl)-N6-(tert-butoxycarbonyl)-L-lysinate (Int-38)

##STR00345##

[0570] Preparation of Int-38: To a stirred solution of Int-35 (750 mg, 1.70 mmol) in CH.sub.2Cl.sub.2 (10.0 mL) was added TEA (0.5 mL, 3.40 mmol) followed by 50% 1-propanephosphonic anhydride solution in EtOAc (2.0 mL, 3.40 mmol). The reaction mixture was for stirred 10 minutes, then Int-26 (974 mg, 1.70 mmol) was added then stirring continued for 15 h (under N.sub.2 atmosphere at ambient temperature). The reaction mixture was diluted with CH.sub.2Cl.sub.2 (20.0 mL) and water (20.0 mL), the organic layer was separated, washed with brine solution (30 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and evaporated under reduced pressure. The obtained crude product was purified by flash column chromatography (silica gel 40 g column) using 4% MeOH in CH.sub.2Cl.sub.2 to afford compound Int-38 (1.30 g, 76%) as an off-white solid. ESI (m/z) [C.sub.58H.sub.69N.sub.5O.sub.10+H].sup.+ 996.

Intermediate 39

Preparation of methyl N2-(3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N6-((S)-2-((tert-butoxycarbonyl)amino)-5-methoxy-5-oxopentanoyl)-L-lysinate (Int-39)

##STR00346##

[0571] Preparation of Int-39: To a solution of amine Int-35 (475 mg, 0.778 mmol), BocNH-Glu(OCH.sub.3)OH (214 mg, 0.817 mmol) and DIPEA (302 mg, 2.335 mmol) in DMF (10 mL) was added HATU (311 mg, 0.817 mmol). The reaction mixture was stirred at room temperature for 2 h. After the solvent was removed, water (30 mL) was added. The white precipitate was filtered and dried, to afford compound Int-39 (603 mg, 95%) as a white solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 7.50-7.47 (m, 4H), 7.29-7.21 (m, 4H), 6.30 (br s, 2H), 6.13 (d, J=7.5 Hz, 1H), 5.30 (br s, 2H), 4.57-4.55 (m, 2H), 4.06 (br s, 1H), 3.71 (s, 3H), 3.66 (s, 3H), 3.17-3.16 (m, 2H), 3.01-2.99 (m, 2H), 2.74-2.34 (m, 8H), 2.10-2.07 (m, 1H), 1.90-1.50 (m, 9H), 1.48 (s, 9H), 1.25-1.20 (m, 2H).

Intermediate 40

Preparation of methyl N6-((S)-2-amino-5-methoxy-5-oxopentanoyl)-N2-(3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysinate (Int-40)

##STR00347##

[0572] Preparation of Int-40: To a solution of Int-39 (300 mg, 0.367 mmol) in THF (5.0 mL) was added 4 N HCl (5.0 mL) at room temperature. The resultant mixture was stirred at room temperature for 3 h. The solvent was removed, and MTBE (30 mL) was added. The solid was collected by filtration and dried, to afford compound Int-40 (259 mg, 94%) as a white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.47 (m, 4H), 7.33-7.22 (m, 8H), 5.05 (s, 2H), 4.38-4.35 (m, 2H), 3.67 (s, 3H), 3.65 (s, 3H), 3.16-3.13 (m, 4H), 2.95-2.93 (m, 2H), 2.66-2.56 (m, 4H), 2.45-2.42 (m, 2H), 2.07-2.03 (m, 2H), 2.01-1.25 (m, 10H). ESI (m/z) [C.sub.40H.sub.52N.sub.4O.sub.8+H].sup.+ 717.

Intermediate 41

Preparation of methyl (E)-N.SUP.., N.SUP..-bis(tert-butoxycarbonyl)-L-argininate (Int-41)

##STR00348##

[0573] Preparation of Int-41a: To a solution of methyl (S)-5-amino-2-(((benzyloxy)carbonyl)amino)pentanoate (3.57 g, 11.2 mmol mmol) in DCM (50 mL) and MeOH (10 mL) was added tert-butyl (E)-(((tert-butoxycarbonyl)amino)(1H-pyrazol-1-yl)methylene)carbamate (3.50 g, 11.2 mmol) and DIPEA (2.91 g, 22.5 mmol). The resultant mixture was stirred at room temperature for 16 h. After concentration, the residue was purified by silica column (30% EtOAc in hexanes), to afford compound Int-41a (4.70 g, 80%) as white solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 8.31 (s, 1H), 7.36-7.31 (m, 5H), 5.53-5.51 (m, 1H), 5.29 (s, 2H), 4.41-4.39 (m, 1H), 3.74 (s, 3H), 3.49-3.38 (m, 2H), 1.91-1.54 (m, 4H), 1.49 (s, 9H), 1.48 (s, 9H); ESI (m/z) [C.sub.25H.sub.38N.sub.4O.sub.8+H].sup.+ 523.

[0574] Preparation of Int-41: To a solution of compound Int-41a (2.00 g, 3.83 mmol) in EtOH (60 mL) was added 20% Pd(OH).sub.2 (250 mg). The resultant mixture was stirred under hydrogen (balloon) at room temperature for 6 h. After filtration, the solvent was removed, to afford compound Int-41 (1.37 g, 92%) as colorless oil: ESI (m/z) [C.sub.17H.sub.32N.sub.4O.sub.6+H].sup.+ 389.

Intermediate 42

Preparation of benzyl (4-(4-(6-aminohexyl)-[1,1-biphenyl]-4-yl)butyl)carbamate (Int-42)

##STR00349##

[0575] Preparation of Int-42a: To a solution of compound Int-19a (100 mg, 0.266 mmol) in pyridine (1.0 mL) was added Tf.sub.2O (90 mg, 0.320 mmol) at 0 C. The reaction mixture was stirred at 0 C. for 1 h and at room temperature for 1 h. Additional Tf.sub.2O (22.5 mg, 0.0800 mmol) was added, and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed, and water (3.0 mL) was added. The precipitated solid was filtered and dried, to afford compound Int-42a (128 mg, 95%) as a white solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 7.61 (d, J=8.5 Hz, 2H), 7.47 (d, J=8.5 Hz, 2H), 7.35-7.19 (m, 9H), 4.88 (s, 2H), 4.73 (br s, 1H), 3.25-3.22 (m, 2H), 2.70-2.65 (m, 2H), 1.68-1.50 (m, 4H).

[0576] Preparation of Int-42b: To a solution of compound Int-42a (150 mg, 0.296 mmol) and 2-(hex-5-yn-1-yl)isoindoline-1,3-dione (101 mg, 0.443 mmol) in dry CH.sub.3CN (3.0 ml) was added triethylamine (120 mg, 1.18 mmol) and copper(I) iodide (2.81 mg, 0.0150 mmol). The solution was degassed with nitrogen for 5 min, then (t-Bu).sub.3P (11.9 mg, 0.059 mmol) and tetrakis (34.2 mg, 0.030 mmol) were added. The resulting reaction mixture was refluxed at 90 C. for 16 h. The solvent was removed, and the residue was purified by silica column (5% to 80% EtOAc in hexanes, product came out at 20% to 40%), to afford compound Int-42b (121 mg, 70%) as a white solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 7.88 (d, J=8.0 Hz, 2H), 7.70 (d, J=8.0 Hz, 2H), 7.49-7.21 (m, 13H), 5.09 (s, 2H), 4.70 (br s, 1H), 3.76 (d, J=7.0 Hz, 2H), 3.23 (q, J=6.5 Hz, 2H), 2.66 (q, J=6.0 Hz, 2H), 2.49 (t, J=7.0 Hz, 2H), 1.90-1.87 (m, 2H), 1.69-1.48 (m, 6H); ESI (m/z) [C.sub.38H.sub.36N.sub.2O.sub.4+H].sup.+ 585.

[0577] Preparation of Int-42c: To a solution of compound Int-42b (120 mg, 0.205 mmol) in EtOH (1.0 mL) and EtOAc (2.0 mL) was added PtO.sub.2 (20 mg). The resultant mixture was stirred under hydrogen (balloon) at room temperature for 3 h. After filtration, the solvent was removed, to afford compound Int-42c (112 mg, 93%) as brown solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 7.80 (d, J=8.0 Hz, 2H), 7.69 (d, J=5.5 Hz, 2H), 7.49-7.46 (m, 4H), 7.35-7.20 (m, 9H), 5.09 (s, 2H), 4.70 (br s, 1H), 3.71 (d, J=6.5 Hz, 2H), 3.24 (q, J=6.5 Hz, 2H), 2.65-2.61 (m, 4H), 1.68-1.40 (m, 12H); ESI (m/z) [C.sub.38H.sub.40N.sub.2O.sub.4+H].sup.+ 589.

[0578] Preparation of Int-42: To a solution of compound Int-42c (110 mg, 0.187 mmol) in MeOH (2.0 mL) and CH.sub.2Cl.sub.2 (2.0 mL) was added hydrazine monohydrate (0.50 mL). The resultant mixture was stirred at room temperature for 3 h. After concentration below 40 C., the residue was partitioned between water (5.0 mL) and CH.sub.2Cl.sub.2 (5.0 mL), the water layer was separated and extracted with CH.sub.2Cl.sub.2 (5.0 mL). The combined organic layers were dried, filtered and concentrated, to afford compound Int-42 (69 mg, 81%) as brown solid: ESI (m/z) [C.sub.30H.sub.38N.sub.2O.sub.2+H].sup.+ 459.

Intermediate 43

Preparation of 5-((4S,4R,4R,5S,5R)-2,2,2,2,2,2-hexamethyl-[4,4:5,4-ter(1,3-dioxolan)]-5-yl)-2-(3-((4S,4R,4R,5S,5R)-2,2,2,2,2,2-hexamethyl-[4,4:5,4-ter(1,3-dioxolan)]-5-yl)propyl)pentan-1-amine (Int-43)

##STR00350## ##STR00351## ##STR00352##

[0579] Preparation of Int-43a: A stirred solution of gluconic acid sodium salt (50 g, 201.6 mmol) in 2,2-dimethoxypropane (300 mL) was charged 4M HCl in MeOH (100 mL) slowly dropwise at room temperature. The reaction mixture was stirred at ambient temperature for 48 h. The reaction mixture was slowly adjusted to pH 6-7 by addition of solid NaHCO.sub.3 then filtered and washed with MeOH (50 mL). The filtrate was evaporated and diluted with CH.sub.2Cl.sub.2 (500 mL), then washed with water (100 mL) and brine (100 mL). The separated organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-43a (25.0 g, 35%) as a light brown oil; ESI (m/z) [C.sub.17H.sub.28O.sub.8+H].sup.+ 361.

[0580] Preparation of Int-43b: To a stirred solution of Int-43a (25 g, 69.4 mmol) in THF (250 mL), was charged with 1M DIBAL in toluene (100 mL, 104 mmol) at 78 C. The resultant reaction mixture warmed to room temperature and stirred for 16 h. The reaction mixture was quenched with saturated NH.sub.4Cl solution, diluted with EtOAc (200 mL) and filtered through celite bed. The filtrate was washed with brine (200 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-43b (17.0 g, 74%) as a light red oil; ESI (m/z) [C.sub.16H.sub.28O.sub.7+H].sup.+ 333.

[0581] Preparation of Int-43c: To a stirred solution of Int-43b (40.0 g, 120.4 mmol) in CH.sub.2Cl.sub.2 (600 mL), was charged Dess-Martin's reagent (61.3 g, 144.5 mmol) at 0 C. The resultant reaction mixture warmed to room temperature and stirred for 6 h. The reaction mixture was quenched with saturated NaHCO.sub.3 (200 mL), the CH.sub.2Cl.sub.2 was separated, and the aqueous layer was extracted with MTBE (3200 mL). The combined organic layers were washed with 10% Na.sub.2S.sub.2O.sub.4 solution (250 mL), followed by brine (100 mL). Organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-43c (45.0 g (crude)) as a light red oil; ESI (m/z) [C.sub.16H.sub.26O.sub.7+H].sup.+ 331.

[0582] Preparation of Int-43d: To a stirred solution of Int-43c (45.0 g, 136.3 mmol) in CH.sub.2Cl.sub.2 (500 mL), was charged PPh.sub.3CHCOOCH.sub.3 (49.5 g, 163.6 mmol) at 0 C. The resultant reaction mixture warmed to room temperature and stirred for 12 h. The reaction mixture was diluted with water (200 mL) and extracted with CH.sub.2Cl.sub.2 (3100 mL). Combined organic layer was washed with brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. Crude material was purified by combi-flash chromatography eluting with 10-30% EtOAc in hexanes to afford Int-43d (30.0 g, 53% over two steps) as a light yellow oil; ESI (m/z) [C.sub.19H.sub.30O.sub.8+H].sup.+ 387.

[0583] Preparation of Int-43e: A stirred solution of Int-43d (2.1 g, 2.57 mmol) in IPA (300 mL) was charged with 10% Pd on carbon 50% wet (500 mg, 50% weight substrate) and stirred under H.sub.2 (balloon) pressure for 3 h. The reaction mixture was filtered through celite bed and concentrated under reduced pressure. Obtained crude compound was purified by combi-flash chromatography eluting with (10-100% EtOAc in hexanes) to afford Int-43e (24 g, 77%) as a liquid; ESI (m/z) [C.sub.19H.sub.32O.sub.8+H].sup.+ 389.

[0584] Preparation of Int-43f: A stirred solution of Int-43e (48 g, 123.7 mmol) in THF (500 mL) was charged with 2M LAH in THF (92 mL, 185.5 mmol) dropwise at 0 C. then gradually allowed to room temperature and stirred for 2 h. The reaction mixture was quenched with saturated Na.sub.2SO.sub.4, diluted with EtOAc (400 mL) filtered through celite bed. The filtrate was washed with brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-43f (43 g, 95%) as a liquid; ESI (m/z) [C18H.sub.32O.sub.7+H].sup.+ 361.

[0585] Preparation of Int-43g: A stirred solution of Int-43f (25 g, 69.4 mmol) in CH.sub.2Cl.sub.2 (300 mL) was charged with PPh.sub.3 (27.2 g, 104.1 mmol), imidazole (23.6 g, 347.2 mmol) and I.sub.2 (26.6 g, 104.1 mmol) then stirred at room temperature for 6 h. The reaction mixture was concentrated under reduced pressure below 30 C. The obtained crude was dissolved in MTBE (500 mL), washed with brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The crude compound was purified by combi-flash chromatography eluting with (10-100% EtOAc in hexanes) to afford Int-43g (19 g, 58%) as a liquid; ESI (m/z) [C.sub.15H.sub.31IO.sub.6+H].sup.+ 471.

[0586] Preparation of Int-43h: A stirred solution of Int-43g (1.8 g, 3.82 mmol) and compound 9 (250 mg, 1.73 mmol) in DMF (15 mL), was charged with K.sub.2CO.sub.3 (716 mg, 5.19 mmol) at ambient temperature and stirred for 16 h. The reaction mixture was diluted with cold water (50 mL) and was extracted with EtOAc (250 mL). The combined organic layer was washed with cold water (320 mL), brine (30 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The obtained crude compound was purified by combi-flash chromatography eluting with 10%-100% EtOAc in hexanes to afford Int-43h (1.0 g, 74%) as a liquid; ESI (m/z) [C.sub.42H.sub.68O.sub.16+H].sup.+ 829.

[0587] Preparation of Int-43i: A stirred solution of Int-43h (2.5 g, 3.82 mmol) in EtOH (30 mL), was charged with 21% NaOEt in EtOH (1.95 mL, 6.03 mmol) at 0 C. The reaction mixture was heated to 90 C. and stirred for 16 h. The reaction mixture was concentrated under reduced pressure. The obtained crude compound was diluted with water (100 mL), acidified to pH=4 with 1N HCl solution and extracted with EtOAc (250 mL). Combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure to afford Int-43i (2.1 g, 95%) as a gummy liquid; ESI (m/z) [C.sub.41H.sub.68O.sub.16+H].sup.+ 817.

[0588] Preparation of Int-43j: A stirred solution of Int-43i (2.1 g, 2.57 mmol) in pyridine (20 mL) was heated at 110 C. and stirred for 3 days. The reaction mixture was concentrated under reduced pressure. Obtained crude compound was purified by combi-flash chromatography eluting with 10-100% EtOAc in hexanes to afford Int-43j (1.2 g, 60%) as a liquid; ESI (m/z) [C.sub.40H.sub.68O.sub.14+H].sup.+ 773.

[0589] Preparation of Int-43k: A stirred solution of Int-43j (1.30 g, 1.68 mmol) in THF (20 mL), was charged with 2M LAH in THF (1.6 mL, 3.36 mmol) at 0 C. then stirred for 2 h. The reaction mixture was quenched with saturated NH.sub.4Cl solution and extracted with EtOAc (250 mL). Combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. Obtained crude compound was purified by combi-flash chromatography eluting with 10-100% EtOAc in hexanes (compound isolated at 20% of EtOAc) to afford Int-43k (1.10 g, 90%) as a gummy liquid; ESI (m/z) [C.sub.38H.sub.66O.sub.13+H].sup.+ 731.

[0590] Preparation of Int-43l: A stirred solution of Int-43k (1.10 g, 1.5 mmol) in CH.sub.2Cl.sub.2 (20 mL), was charged with Et.sub.3N (0.6 mL, 4.5 mmol), followed by MsCl (0.153 mL, 2.26 mmol) at 0 C. then stirred for 2 h. The reaction mixture was diluted with CH.sub.2Cl.sub.2 (50 mL), washed with water (20 mL), saturated NaHCO.sub.3 solution (30 mL), brine (30 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-43l (1.20 g, 98%) as a colour less gum; ESI (m/z) [C.sub.39H.sub.68O.sub.15S+H].sup.+ 809.

[0591] Preparation of Int43m: To a stirred solution of compound Int-43l (0.51 g, 0.63 mmol) in DMF (10 mL) was added sodium azide (238 mg, 3.7 mmol) at room temperature. The reaction mixture was warmed to 60 C. and stirred for 2 h. The reaction mixture was quenched with ice water (20 mL) and extracted with EtOAc (3200 mL). Combined EtOAc layers were dried over anhydrous Na.sub.2SO.sub.4, filtered and evaporated to afford compound Int-43m (0.52 g, crude) as a yellow liquid: ESI (m/z) [C.sub.38H.sub.67N.sub.3O.sub.12+H].sup.+ 759.

[0592] Preparation of Int-43: To a stirred solution of compound Int-43m (0.52 g, 0.70 mmol) in THF:water (6:2) was added triphenylphosphine (185 mg, 0.70 mmol). The reaction mixture was stirred for 16 h at room temperature. After completion of the reaction, the reaction mixture was evaporated under reduced pressure to get crude material which was purified by combi flash (120 g column) chromatography eluting with 10% EtOAc in hexanes to afford compound Int-43 (0.49 g, 98%) as an off-white solid. ESI (m/z) [C.sub.38H.sub.67NO.sub.12+H].sup.+ 730.

Intermediate 44

Preparation of tert-butyl (E)-((3-amino-5H-pyrrolo[2,3-b]pyrazine-2-carboxamido)(methylthio)methylene)carbamate (Int-44)

##STR00353##

[0593] Preparation of (Int-44): To a stirred solution of 3-amino-5H-pyrrolo[2,3-b]pyrazine-2-carboxylic acid (WO2018096325 A1, 1.0 g, 5.5 mmol) in DMF (20.0 mL) was added DIPEA (3.21 g, 24.96 mmol) followed by HATU (3.30 g, 8.62 mmol). The reaction mixture was stirred for 15 min. 1-N-Boc-2-Methyl-Isothiourea (1.60 g, 8.33 mmol) was added then stirred at room temperature for 12 h. The reaction mixture was diluted with EtOAc (100.0 mL), washed with water (50 mL) and brine (50 mL). The Organic layer was separated, dried over Na.sub.2SO.sub.4, filtered and volatiles were removed under reduced pressure. The crude product was purified by combi flash chromatography eluting with 50-60% EtOAc/hexanes to afford compound Int-44 (m/z) [C.sub.14H.sub.18N.sub.6O.sub.3S+H].sup.+ 351.

Intermediate 45

Preparation of di-tert-butyl (6-((3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propyl)amino)-6-oxohexane-1,5-diyl)(S)-dicarbamate (Int-45)

##STR00354##

[0594] Preparation of Int-45: To a stirred solution of Int-17 (290 mg, 0.84 mmol) in dichloromethane (20.0 mL) was added DIPEA (325 mg, 2.52 mmol) followed by T.sub.3P (50% in EtOAc) (800 mg, 3.48 mmol). The reaction mixture was stirred for 15 mins, N-Boc-Lys(Boc)-OH (350 mg, 0.84 mmol) was added then stirred at room temperature for 3 h. After completion, the reaction mixture was concentrated under reduced pressure to afford crude which was purified by combi-flash chromatography eluting with (75% EtOAc in hexanes) to afford Int-45 (300 mg, 51%) as an off-white solid; ESI (m/z) [C.sub.43H.sub.60N.sub.4O.sub.7+H].sup.+ 745.

Intermediate 46

Preparation of methyl N.SUP.6.-(tert-butoxycarbonyl)-N.SUP.2.-methyl-L-lysinate (Int-46)

##STR00355##

[0595] Preparation of Int-46a: A solution of Lys(Boc)-OCH.sub.3 (2.50 g, 9.60 mmol) and benzaldehyde (1.03 g, 9.60 mmol) in MeOH (40 mL) stirred for 15 min. NaCNBH.sub.3 (0.90 g, 14.4 mmol) was added at 0 C. then the reaction mixture was warmed to ambient temperature and stirred for 4 h. The volatiles were removed under reduced pressure and the obtained crude compound used to next step without purification Int-46a (2.10 g, crude). ESI (m/z) [C.sub.19H.sub.30N.sub.2O.sub.4+H].sup.+ 351.

[0596] Preparation of Int-46b: A solution of Int-46a (2.10 g, 6.0 mmol) and formaldehyde (10.0 mL, 37% solution in water) in MeOH (20 mL) was stirred for 15 min. NaCNBH.sub.3 (0.56 g, 9.0 mmol) was added at 0 C. then the reaction mixture was warmed to ambient temperature and stirred for 4 h. The volatiles were removed under reduced pressure, reaction mixture was diluted with water (30 mL) and extracted with EtOAc (250 mL). The combined organic layer was washed with brine solution, dried over anhydrous Na.sub.2SO.sub.4, filtered, and evaporated under reduced pressure to obtain crude product which was purified by flash column chromatography (silica gel 40 g column) using 15-20% EtOAc in hexanes to afford compound Int-46b (0.90 g, 42%) as an off-white solid. ESI (m/z) [C.sub.20H.sub.32N.sub.2O.sub.4+H].sup.+ 365.

[0597] Preparation of Int-46: To a stirred solution of Int-46b (900 mg, 2.47 mmol) in IPA (15.0 mL) was added Pd on carbon (10%, 50% wet, 150 mg) at room temperature under nitrogen atmosphere. The nitrogen was replaced with a H.sub.2 balloon, and the reaction mixture was stirred under 1 atm of hydrogen for 40 minutes. The reaction mixture was filtered through celite pad and washed with IPA (40 mL). The filtrate was concentrated to afford compound Int-46 (600 mg, 88%) as a colorless gum. ESI (m/z) [C.sub.13H.sub.26N.sub.2O.sub.4+H].sup.+ 275.

Intermediate 47

Preparation of methyl (E)-3-(3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acrylate (Int-47)

##STR00356##

[0598] Preparation of Int-47a: To a stirred solution of Int-2 (2.0 g, 5.03 mmol) and Int-7 (4.06 g, 12.6 mmol) in a mixture of DMF/H.sub.2O (30 mL:6 mL) was added Na.sub.2CO.sub.3 (1.60 g, 15.09 mmol) at room temperature. The reaction mixture was degassed for 20 min with Argon then Pd(PPh.sub.3).sub.4 (63 mg, 0.05 mmol) was added. The reaction mixture was warmed to 100 C. and stirred for 4 h. The reaction mixture was cooled to room temperature, water (50 ml) was added then extracted with EtOAc (250 mL). Combined organic layer was washed with cold water (320 mL) followed by brine solution (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The obtained crude material was purified by combi-flash chromatography eluting with (1%-100% EtOAc in hexanes) to afford Int-47a (0.70 g, 32%) as a white solid. ESI (m/z) [C.sub.25H.sub.30ClNO.sub.4+H].sup.+ 444.

[0599] Preparation of Int-47: To a stirred solution of compound Int-47a (0.80 g, 1.8 mmol) in IPA (10 mL) and CH.sub.3COOH (0.2 mL), was added PtO.sub.2 (100 mg). Reaction mixture stirred under H.sub.2 balloon pressure for 3 h. The reaction mixture was filtered through celite, and filtrate was concentrated under reduced pressure to afford Int-47 (700 mg, 87%) as an off-white slid. ESI (m/z) [C.sub.25H.sub.32ClNO.sub.4+H].sup.+ 446.

Intermediate 48

Preparation of benzyl (4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)butyl)carbamate (Int-48)

##STR00357##

[0600] Preparation of Int-48: To a stirred solution of compound Int-1 (5.0 g, 11.59 mmol) in 1,4-dioxane (50 mL) was added KOAc (2.27 g, 23.19 mmol) followed by bis-pincalate diborane (3.50 g, 13.91 mmol) at room temperature. The reaction mixture was degassed with nitrogen for 5 min, and PdCl.sub.2(dppf) (0.42 g, 0.57 mmol) was added. The reaction mixture was again degassed with nitrogen for 2 min then warmed to 100 C. and stirred for 16 h. The reaction mixture was filtered through celite pad, washed with EtOAc (100 mL) and filtrate was concentrated under reduced pressure to get crude material which was purified by flash chromatography eluting with (0-15% EtOAc in hexanes) to afford compound Int-48 (4.0 g, 84%) as a colorless liquid. ESI (m/z) [C.sub.24H.sub.32BNO.sub.4+H].sup.+ 410.

Intermediate 49

Preparation of tert-butyl (4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)butyl)carbamate (Int-49)

##STR00358##

[0601] Preparation of Int-49: To a stirred solution of Int-2 (3.0 g, 7.55 mmol) in 1,4-dioxane (100 mL) was added KOAc (1.40 g, 15.10 mmol) followed by bis pincalate diborane (2.30 g, 9.06 mmol) at room temperature. The reaction mixture was degassed with nitrogen for 5 min, then PdCl.sub.2(dppf) (0.28 g, 0.38 mmol) was added. The reaction mixture was again degassed with nitrogen for 2 min then warmed to 100 C. and stirred for 16 h, The reaction mixture was filtered through celite pad, washed with EtOAc and filtrate was concentrated. The obtained crude product was purified by flash chromatography eluting with (0-25% EtOAc in hexane) to afford compound Int-49 (2.60 g, 88%) as a colorless liquid. ESI (m/z) [C.sub.21H.sub.34BNO.sub.4+H].sup.+ 376.

Intermediate 50

Preparation of tert-butyl (S)-(5-(3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanamido)-5-(1H-tetrazol-5-yl)pentyl)carbamate (Int-50)

##STR00359##

[0602] Preparation of Int-50a: A stirred solution of Int-35a (1.20 g, 1.78 mmol) in MeOH (5 mL), was charged with 7M NH.sub.3 in MeOH (30 mL) at room temperature and stirred for 64 h. The reaction mixture was concentrated under reduced pressure. The obtained solid compound was washed with 50% MTBE in hexanes (220 mL) and dried under reduced pressure to afford Int-50a (0.85 g, 72%) as an off-white solid: ESI (m/z) [C.sub.38H.sub.50N.sub.4O.sub.6+H].sup.+ 659.

[0603] Preparation of Int-50b: A solution of Int-50a (1.1 g, 1.67 mmol) in THF (20 mL), was charged with Et.sub.3N (0.70 mL, 5.01 mmol) followed by TFAA (0.58 mL, 4.17 mmol) slowly dropwise at 0 C. The reaction mixture stirred at room temperature for 16 h. The reaction mixture was diluted with water and extracted with EtOAc (350 mL). The combined organic extracts were concentrated under reduced pressure. The obtained crude solid was treated with K.sub.2CO.sub.3 (691 mg, 5.01 mmol) in MeOH (30 mL) at room temperature for 15 minutes then water (50 mL) added and extracted with CH.sub.2Cl.sub.2 (350 mL). The separated organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-50b (900 mg, 84%) as an off-white solid; ESI (m/z) [C.sub.38H.sub.48N.sub.4O.sub.5+H].sup.+ 641.

[0604] Preparation of Int-50: A solution of Int-50b (900 mg, 1.40 mmol) in DMF (20 mL), was charged with NaN.sub.3 (274 mg, 4.21 mmol) followed by NH.sub.4Cl (188 mg, 3.51 mmol). The reaction mixture was warmed to 100 C. and stirred for 24 h. The reaction mixture was diluted with water, acidified to pH=3 with 10% citric acid solution and filtered. The solid was washed with water followed by MTBE then dried under reduced pressure to afford Int-50 (750 mg, 78%) as an off-white solid; ESI (m/z) [C.sub.38H.sub.49N.sub.7O.sub.5+H].sup.+ 684

Intermediate 51

Preparation of tert-butyl (S)-(5-(3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)-N-methylpropanamido)-5-(1H-tetrazol-5-yl)pentyl)carbamate (Int-51)

##STR00360##

[0605] Preparation of Int-51a: To a solution of Int-23 (300 mg, 0.695 mmol) in DMF (3.0 mL) was added HATU (291 mg, 0.765 mmol) followed by compound Int-46 (200 mg, 0.730 mmol) and DIPEA (270 mg, 2.086 mmol). The reaction mixture was stirred at rt for 1 h. The reaction mixture was partitioned with water (30 mL) and EtOAc (30 mL). The water phase was extracted with EtOAc (30 mL), and the combined EtOAc layers were concentrated, to afford compound Exp-51a (452 mg, 95%) as white solid: ESI (m/z) [C.sub.40H.sub.53N.sub.3O.sub.7+H].sup.+ 688.

[0606] Preparation of Int-51b: A stirred solution of Int-51a (1.20 g, 1.74 mmol) in MeOH (5 mL), was charged with 7M NH.sub.3 in MeOH (30 mL) at room temperature then stirred for 64 h. The reaction mixture was concentrated under reduced pressure. Crude material was purified by combi-flash chromatography eluting with 1-10% MeOH in dichloromethane (compound isolated at 5% of MeOH) to afford Int-51b (750 mg, 65%) as an off-white solid. ESI (m/z) [C.sub.39H.sub.52N.sub.4O.sub.6+H].sup.+ 673.

[0607] Preparation of Int-51c: A solution of Int-51b (750 mg, 1.11 mmol) in THF (15 mL), was charged with Et.sub.3N (0.460 mL, 3.33 mmol) followed by TFAA (0.393 mL, 2.79 mmol) slowly dropwise at 0 C. The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with water and extracted with EtOAc (3100 mL) and concentrated under reduced pressure. The obtained crude solid was treated with K.sub.2CO.sub.3 (459 mg, 3.33 mmol) in MeOH (50 mL) at room temperature for 15 minutes then water (100 mL) added. The reaction mixture was extracted with dichloromethane (3100 mL). The combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. Crude material was purified by combi-flash chromatography eluting with 1-100% EtOAc in hexanes to afford Int-51c (600 mg, 82%) as a yellow solid; ESI (m/z) [C.sub.39H.sub.50N.sub.4O.sub.5+H].sup.+ 655.

[0608] Preparation of Int-51: A solution of Int-51c (600 mg, 0.92 mmol) in DMF (10 mL) was charged with NaN.sub.3 (178 mg, 2.75 mmol) followed by NH.sub.4Cl (122.4 mg, 2.29 mmol). The reaction mixture was warmed to 100 C. and stirred for 24 h. The reaction mixture was diluted with water, acidified to pH=3 with 10% citric acid solution then washed with EtOAc (350 mL). The combined organic layer was washed with cold water (320 mL), brine solution (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. Crude material was purified by combi-flash chromatography eluting with 1-10% MeOH in dichloromethane to afford compound Int-51 (400 mg, 62%) as light yellow semisolid; ESI (m/z) [C.sub.39H.sub.51N.sub.7O.sub.5+H].sup.+ 698.

Intermediate 52

Preparation of methyl 3-(3-(4-((tert-butoxycarbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanoate (Int-52)

##STR00361##

[0609] Preparation of Int-52a: To a solution of 3-(hydroxylphenyl)boronic acid (10.0 g, 41.13 mmol) and methyl 3-(4-bromophenyl)propanoate (5.63 g, 41.13 mmol) in a mixture of DMF/H.sub.2O (100 mL:30 mL) was added K.sub.3PO.sub.4 (17.4 g, 82.2 mmol) at room temperature. The reaction mixture was degassed for 10 min with Argon then Pd(PPh.sub.3).sub.4 (1.42 g, 1.23 mmol) was added. The reaction mixture was warmed to 100 C. and stirred for 16 h. The reaction mixture was cooled to room temperature, filtered through celite bed and washed with EtOAc (500 mL). The filtrate was washed with cold water (320 mL), brine solution (30 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The obtained crude was purified by combi flash column chromatography eluting with 1%-100% EtOAc in hexanes (compound isolated at 60% of EtOAC) to afford Int-52a (3.0 g, 29%) as an off-white solid: ESI (m/z) [C.sub.16H.sub.16O.sub.3+H].sup.+ 256.

[0610] Preparation of Int-52b: To a solution of Int-51a (3.0 g, 11.7 mmol) in pyridine (30 mL) was added triflic anhydride (2.90 mL, 17.5 mmol) at 0 C. The reaction mixture was warmed to room temperature and stirred for 2 h. The reaction mixture was poured into a beaker containing cold water (100 mL) then extracted with EtOAc (350 mL). The combined organic layer was washed with 2N HCl solution (320 mL), brine solution (20 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford compound Int-52b (3.80 g, 84%) as a brown liquid: ESI (m/z) [C.sub.17H.sub.15F.sub.3O.sub.5S+H].sup.+ 389.

[0611] Preparation of Int-52c: To a solution of Int-52b (3.80 g, 9.79 mmol) and tert-butyl but-3-yn-1-ylcarbamate (2.15 g, 12.7 mmol) in CH.sub.3CN (50 mL), was added bis(triphenylphosphine)palladium(II) chloride (0.430 g, 0.613 mmol) and t-Bu.sub.3P (10% in hexanes, 395 mg, 1.95 mmol). The reaction mixture was degassed with nitrogen for 10 min and charged with triethylamine (5.50 mL, 39.16 mmol) followed by copper(I) iodide (372 mg, 1.95 mmol). The reaction mixture warmed to 80 C. and stirred for 16 h. The reaction mixture was diluted with EtOAc (200 mL), filtered through celite bed and the filtrate was washed with water (50 mL) followed by brine solution (50 mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The crude was purified by combi-flash column chromatography eluting with 1%-80% EtOAc in hexanes (compound isolated at 50% of EtOAc) to afford compound Int-52c (2.60 g, 65%) as a brown solid: ESI (m/z) [C.sub.25H.sub.29NO.sub.4+H].sup.+ 408.

[0612] Preparation of Int-52: To a solution of Int-52c (2.20 g, 5.4 mmol) in EtOH (30 mL) and EtOAc (30 mL), was added 20% Pd(OH).sub.2 on carbon 50% wet (400 mg, 50% weight substrate, weight by weight) under an inert atmosphere. The reaction mixture was stirred under H.sub.2 (100 psi) pressure in an autoclave for 16 h. The reaction mixture was filtered through celite bed then the filtrate was concentrated under reduced pressure. The obtained crude was purified by combi-flash column chromatography eluting with 1%-100% EtOAc in hexanes (compound isolated at 40% of EtOAc) to afford compound Int-52 (1.40 g, 61%) as an off-white solid: ESI (m/z) [C.sub.25H.sub.33NO.sub.4+H].sup.+ 412.

Intermediate 53

Preparation of methyl (3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-valinate (Int-53)

##STR00362##

[0613] Preparation of Int-53: To a stirred solution of Int-23 (2.00 g, 4.64 mmol) and HATU (2.11 g, 5.56 mmol) in DMF (20 mL) was added Val-OCH.sub.3 (0.929 g, 5.56 mmol) followed by DIPEA (2.3 mL, 13.9 mmol). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with cold water (50 mL) and the precipitated solid was collected by filtration. The solid was dissolved in EtOAc (150 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Int-53 (2.1 g, 83%) as an off-white solid: ESI (m/z) [C.sub.33H.sub.40N.sub.2O.sub.5+H].sup.+ 545.

Intermediate 54

Preparation of methyl N2-(3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N6-(tert-butoxycarbonyl)-N2-(2-((tert-butoxycarbonyl)amino)ethyl)-L-lysinate (Int-54)

##STR00363##

[0614] Preparation of Int-54a: To a solution of tert-butyl (2-hydroxyethyl)carbamate (2.0 g, 12.41 mmol) in CH.sub.2Cl.sub.2 (40 mL) was added Dess-Martin reagent at 0 C. The resultant solution was stirred at rt for 2 h. After the solvent was removed, the residue was partitioned in MTBE (50 mL) and 2 N NaHCO.sub.3 (50 mL), and the mixture was stirred at rt for 10 min. The white solid was filtered off, and MTBE layer was separated, dried and concentrated, to afford compound Int-54a (1.73 g, 88%) as a white solid: ESI (m/z) [C.sub.7H.sub.13NO.sub.3+H].sup.+ 160.

[0615] Preparation of Int-54b: To a solution of Lys(Boc)-OCH.sub.3 (2.00 g, 6.74 mmol) in MeOH (40 mL) was added Int-54a (1.07 g, 6.74 mmol), AcOH (405 mg, 6.74 mmol) and NaCNBH.sub.3 (423 mg, 6.74 mmol). The resulting solution was stirred at rt for 16 h. After the solvent was removed, the residue was purified by silica column (5% to 60% of EtOAc in CH.sub.2Cl.sub.2, product came out at 10% to 30%), to afford Int-54b (1.25 g, 46%) as a yellow oil: ESI (m/z) [C.sub.19H.sub.37N.sub.3O.sub.6+H].sup.+ 404.

[0616] Preparation of Int-54: To a solution of Int-54b (1.02 g, 2.55 mmol) in DMF (20 mL) was added Int-23 (1.10 g, 2.55 mmol), HATU (1.16 g, 3.06 mmol) and DIPEA (988 mg, 7.65 mmol). The resulting solution was stirred at rt for 48 h. Additional Int-23 (204 mg, 0.510 mmol), HATU (193 mg, 0.510 mmol) and DIPEA (197 mg, 1.53 mmol) were added, and the resultant solution was heated at 45 C. for 6 h. After the solvent was removed, water (20 mL) was added. The resulting white solid was collected by filtration, and purified by silica column (0% to 30% of EtOAc in CH.sub.2Cl.sub.2, product came out at 10%), to afford Int-54 (1.26 g, 60%) as a white solid: ESI (m/z) [C.sub.46H.sub.64N.sub.4O.sub.9+H].sup.+ 817.

Intermediate 55

Preparation of methyl tert-butyl N.SUP.2.-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)ethyl)-N.SUP.2.-(3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.-(tert-butoxycarbonyl)-L-lysinate (Int-55)

##STR00364##

[0617] Preparation of Int-55a: To a solution of 2-aminoethan-1-ol (2.00 g, 32.7 mmol) and NaHCO.sub.3 (4.13 g, 49.1 mmol) in THF (30 mL), MeOH (30 mL) and water (10 mL) was added FmocCl (8.47 g, 32.7 mmol) at 0 C. The reaction mixture was warmed to rt and stirred for 3 h. After the solvent was removed, water (200 mL) was added. The solid was collected by filtration and dried by lyophilization, to afford Int-55a (8.66 g, 93%) as a white solid: ESI (m/z) [C.sub.17H.sub.17NO.sub.3+H].sup.+ 284.

[0618] Preparation of Int-55b: To a solution of Int-55a (2.00 g, 7.06 mmol) in CH.sub.2Cl.sub.2 (60 mL) was added Dess-Martin periodinane (3.59 g, 8.47 mmol) at 0 C. The reaction mixture was warmed to rt and stirred for 3 h. After the solvent was removed, the residue was partitioned with MTBE (100 mL) and NaHCO.sub.3 (100 mL). The bi-phase system was stirred at rt for 10 min. The solid was filtered off and the MTBE layer was collected by separation funnel. The water layer was washed with MTBE (20 mL) and the organic layers were combined, dried and concentrated, to afford Int-55b (1.85 g, 93%) as a yellow oil: ESI (m/z) [C.sub.17H.sub.15NO.sub.3+H].sup.+ 282.

[0619] Preparation of Int-55c: To a solution of Lys(Boc)Ot-Bu (100 mg, 0.295 mmol) and Int-55b (83 mg, 0.295 mmol) in MeOH (3.0 mL) was added NaCNBH.sub.3 (22.2 mg, 0.354 mmol) and AcOH (21.2 mg, 0.354 mmol) at rt. The reaction mixture was stirred for 16 h. After the solvent was removed to the resulting residue was added CH.sub.2Cl.sub.2 (20 mL). The solid was filtered off and washed with CH.sub.2Cl.sub.2 (10 mL). The CH.sub.2Cl.sub.2 filtrate was concentrated and purified by silica column (10% to 80% of EtOAc in CH.sub.2Cl.sub.2, product came out at 40% to 60%), to afford Int-55c (103 mg, 61%) as yellow oil: ESI (m/z) [C.sub.32H.sub.45N.sub.3O.sub.6+H].sup.+ 568.

[0620] Preparation of Int-55: To a solution of Int-55c (132 mg, 0.232 mmol) and Int-23 (100 mg, 0.232 mmol) in DMF (3.0 mL) was added HATU (106 mg, 0.278 mmol) and DIPEA (90.0 mg, 0.695 mmol) at rt. The reaction mixture was stirred at rt for 16 h. After the solvent was removed, the residue was purified by silica column (0% to 50% of EtOAc in CH.sub.2Cl.sub.2, product came out at 10%), to afford compound Int-55 (151 mg, 66%) as a white solid: ESI (m/z) [C.sub.59H.sub.72N.sub.4O.sub.9+H].sup.+ 981.

Intermediate 56

[0621] Preparation of tert-butyl (S)-(5-(N-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)ethyl)-3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanamido)-5-(1H-tetrazol-5-yl)pentyl)carbamate (Int-56)

##STR00365##

[0622] Preparation of Int-56a: To a stirred solution of Lys(Boc)-OCH.sub.3 (5.0 g, 16.8 mmol) in THF (60 mL) was added saturated NaHCO.sub.3 solution (8.8 mL) followed by 50% CbzCl in toluene (8.6 mL, 25.3 mmol) at 0 C. The resulted reaction mixture was gradually allowed to room temperature and stirred for 16 h. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (330 mL). The combined organic layer was washed with brine solution (50 mL) and dried over anhydrous Na.sub.2SO.sub.4, filtered and volatiles were removed under reduced pressure. The obtained crude was purified by combi-flash chromatography eluting with 10-50% EtOAc in hexanes (compound isolated at 30% of EtOAc) to afford Int-56a (5.5 g, 83%) as a colourless liquid; ESI (m/z) [C.sub.20H.sub.30N.sub.2O.sub.6+H].sup.+ 395.

[0623] Preparation of Int-56b: A stirred solution of Int-56a (5.5 g, 13.9 mmol) in MeOH (5 mL) was charged with 7M NH.sub.3 in MeOH (50 mL) at room temperature then stirred for 24 h. The reaction mixture again charged with 7M NH.sub.3 in MeOH (10 mL) and stirring continued for 24 h. The reaction mixture was concentrated under reduced pressure. The obtained solid compound was washed with 10% MTBE in hexanes (220 mL) and dried under reduced pressure to afford Int-56b (4.4 g, 83%) as a white solid; ESI (m/z) [C.sub.19H.sub.29N.sub.3O.sub.5+H].sup.+ 380

[0624] Preparation of Int-56c: To a solution of Int-56b (4.40 g, 11.6 mmol) in THF (50 mL) was added Et.sub.3N (4.88 mL, 34.8 mmol) followed by TFAA (4.09 mL, 29.02 mmol) slowly dropwise at 0 C. The reaction mixture was warmed to room temperature and stirred for 16 h. The reaction mixture was diluted with water (100 mL), extracted with EtOAc (350 mL) then the organic layer was concentrated under reduced pressure. The obtained crude solid was treated with K.sub.2CO.sub.3 (4.80 g, 34.8 mmol) in MeOH (50 mL) at room temperature for 15 minutes then water (100 mL) added followed by extraction with dichloromethane (350 mL). The separated organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford compound Int-56c (4.0 g, 95%) as a light yellow solid; ESI (m/z) [C.sub.19H.sub.27N.sub.3O.sub.4+H].sup.+ 362

[0625] Preparation of Int-56d: To a solution of Int-56c (3.5 g, 9.69 mmol) in DMF (30 mL) was added NaN.sub.3 (1.89 g, 29.0 mmol) followed by NH.sub.4Cl (1.30 g, 24.1 mmol). The reaction mixture was warmed to 100 C. and stirred for 16 h. The reaction mixture was diluted with water (50 mL), acidified to pH=3 with 10% citric acid solution and extracted with EtOAc (350 mL). Combined organic layer was washed with cold water (320 mL), brine solution (40 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The obtained solid washed with MTBE (20 mL) and dried to afford Int-56d (1.50 g, 38%) as a brown solid; ESI (m/z) [C.sub.19H.sub.28N.sub.6O.sub.4+H].sup.+ 405

[0626] Preparation of Int-56e: To a solution of Int-56d (2.9 g, 7.17 mmol) in IPA (20 mL) and EtOAc (20 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (400 mg, 50% weight substrate) under inert atmosphere. The resultant reaction mixture stirred under H.sub.2 balloon pressure for 3 h. The reaction mixture was filtered through celite bed and filtrate was concentrated under reduced pressure. The obtained crude solid was stirred in CH.sub.3CN (10 mL), filtered, washed with CH.sub.3CN (10 mL) then dried to afford compound Int-56e (0.90 g, 46%) as an off-white solid; ESI (m/z) [C.sub.11H.sub.22N.sub.6O.sub.2+H].sup.+ 271.

[0627] Preparation of Int-56f: To a solution of compound Int-56e (600 mg, 2.21 mmol) and Int-55b (624 mg, 2.21 mmol) in MeOH (20 mL) was charged with AcOH (172 mg, 2.88 mmol) followed by NaBH.sub.3CN (178 mg, 2.88 mmol). The resultant reaction mixture stirred at room temperature for 3 h. The reaction mixture was concentrated under reduced pressure. Crude material was purified by reverse phase combi flash chromatography eluting with 10-100% CH.sub.3CN in water (compound isolated at 40% of CH.sub.3CN) to afford compound Int-56f (250 mg, 21%) as white solid; ESI (m/z) [C.sub.28H.sub.37N.sub.7O.sub.4+H].sup.+ 536.

[0628] Preparation of Int-56: To a stirred solution of Int-23 (225 mg, 0.523 mmol) and Int-56f (280 mg, 0.523 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added Et.sub.3N (0.29 mL, 2.09 mmol) followed by 50% T.sub.3P in EtOAc (0.49 mL, 0.78 mmol). The reaction mixture was stirred at room temperature for 3 h. The reaction mixture was concentrated under reduced pressure. Water (50 mL) was added to the obtained crude and the aqueous solution was washed with EtOAc (350 mL). The combined EtOAc layer was washed with brine solution (50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The solid was washed with MTBE (210 mL) and dried to afford Int-56 (400 mg, 80%) as a brown solid; ESI (m/z) [C.sub.55H.sub.64N.sub.8O.sub.7+H].sup.+ 949.

Intermediate 57

Preparation of methyl N.SUP.2.-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propyl)-N.SUP.2.-(3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.-(tert-butoxycarbonyl)-L-lysinate (Int-57)

##STR00366##

[0629] Preparation of Int-57: Int-57 was prepared beginning with Lys(Boc)-OCH.sub.3 and 3-aminopropanol following an analogous series of procedures to the synthesis of Int-55.

Intermediate 58

[0630] Preparation of tert-butyl (S)-(5-(N-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propyl)-3-(4-(4-(((benzyloxy)carbonyl)amino)butyl)-[1,1-biphenyl]-4-yl)propanamido)-5-(1H-tetrazol-5-yl)pentyl)carbamate (Int-58)

##STR00367##

[0631] Preparation of Int-58: Int-58 was prepared substituting in (9H-fluoren-9-yl)methyl (3-oxopropyl)carbamate and following an analogous series of procedures to the synthesis of Int-56.

Intermediate 59

Preparation of (2R,2R,3R,3R,4R,4R,5S,5'S)-6,6-((3-(4-aminophenyl)propyl)azanediyl) bis(hexane-1,2,3,4,5-pentaol) (Int-59)

##STR00368##

[0632] Preparation of Int-59a: To a stirred solution of 3-(4-nitrophenyl)propan-1-amine hydrochloride (2.0 g, 9.25 mmol) in MeOH (100 mL), was added D(+)glucose (8.3 g, 46.29 mmol), AcOH (2.7 g, 46.27 mmol) and NaCNBH.sub.3 (2.91 g, 46.29 mmol) at room temperature. The resultant reaction mixture heated to 65 C. and stirred for 12 h. The volatiles were removed under reduced pressure to obtain the crude material. The crude material was diluted with 4M HCl in MeOH (20 mL) and stirred for 45 minutes, concentrated under reduced pressure then azeotroped with MeOH (410 mL). Obtained crude was purified by C-18 reverse phase column chromatography using 40-60% CH.sub.3CN in H.sub.2O to afford Int-59a (3.0 g, 62%) as an off-white gummy solid; ESI (m/z) [C.sub.21H.sub.36N.sub.2O.sub.12+H].sup.+ 509.

[0633] Preparation of Int-59: To a stirred solution of Int-59a (1.4 g, 2.75 mmol) in MeOH/H.sub.2O (50.0 mL, 10:1) was added to 10% Pd/C 50% wet (200 mg, 50% weight substrate) at room temperature under nitrogen atmosphere. The nitrogen atmosphere was replaced with H.sub.2 (balloon) and the reaction mixture was stirred for 12 h. The reaction mixture was filtered through celite pad, washed with IPA and H.sub.2O (20 mL, 1:1) then the filtrate was concentrated under reduced pressure to afford Int-59 (1.1 g, crude) as an off-white gummy solid; ESI (m/z) [C.sub.21H.sub.38N.sub.2O.sub.10+H].sup.+ 479.

Intermediate 60

Preparation of ((2R,3R,4R,5S)-6-((2-aminoethyl) ((2R,3R,4R,5R)-2,3,4,5,6-pentahydroxy hexyl)amino)hexane-1,2,3,4,5-pentaol.Math.2HCl salt; (Int-60)

##STR00369##

[0634] Preparation of Int-60a: A solution of tert-butyl (2-aminoethyl)carbamate (30.0 g, 156.2 mmol) in MeOH (300 mL) was charged with benzaldehyde (19.8 g, 187.5 mmol) and AcOH (11.2 g, 187.5 mmol) at room temperature and the resultant reaction mixture was stirred for 16 h. The reaction mixture was charged with NaBH.sub.3CN (13.9 g, 225.0 mmol) continued at room temperature for another 24 h. The reaction mixture was concentrated under reduced pressure, obtained crude was diluted with EtOAc (250 mL) then washed with saturated NaHCO.sub.3 (3150 mL), brine (100 mL). Separated organic layer dried over NaSO.sub.4 and concentrated under reduced pressure. Crude was purified by combi-flash chromatography eluted with 1-10% MeOH in dichloromethane, compound isolated at 6% of MeOH to compound Int-60a (26.0 g, 55%) as a light yellow liquid: ESI (m/z) [C.sub.14H.sub.22N.sub.2O.sub.2+H].sup.+ 251.

[0635] Preparation of Int-60b: A solution of Int-60a (26.0 g, 88.0 mmol) and D(+)glucose (37.4 g, 208.0 mmol) in MeOH (260 mL) was charged with AcOH (12.2 mL, 208 mmol) followed by NaBH.sub.3CN (12.89 g, 208.0 mmol), the resultant reaction mixture was stirred at 60 C. for 16 h. The reaction mass was concentrated under reduced pressure, obtained crude was purified by reverse phase combi flash purification eluted with 10-100% CH.sub.3CN in H.sub.2O (compound isolated at 40% of CH.sub.3CN) to afford compound Int-60b (38.0 g, 88%) as a colourless gum: ESI (m/z) [C.sub.20H.sub.34N.sub.2O.sub.7+H].sup.+ 415.

[0636] Preparation of compound Int-60c: To a solution Int-60b (38.0 g, 91.7 mmol) in IPA (190 mL) and H.sub.2O (190 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (3.8 g, 50% weight substrate) under inert atmosphere. The resultant reaction mixture stirred under hydrogen atmosphere (200 psi, autoclave) for 16 h. The reaction mass filtered through celite bed and filtrate was concentrated under reduced pressure to afford compound Int-60c (28.0 g, 94%) as a colourless gum; ESI (m/z) [C.sub.13H.sub.28N.sub.2O.sub.7+H].sup.+ 325.

[0637] Preparation of compound Int-60d: A solution of Int-60c (30 g, 92.6 mmol) and D(+)mannose (24.9 g, 138.8 mmol) in MeOH (300 mL) was charged with AcOH (8.30 mL, 138.8) followed by NaBH.sub.3CN (4.3 g, 138.8 mmol), the resultant reaction mixture was stirred at 60 C. for 16 h. The reaction mixture was cooled to room temperature, charged with D(+)mannose (8.33 g, 46.3 mmol) and NaBH.sub.3CN (2.87 g, 46.3 mmol). The resultant reaction heated at 60 C. and stirred for 24 h. The reaction mass was concentrated under reduced pressure, obtained crude was purified by reverse phase combi flash purification eluted with 10-100% CH.sub.3CN in H.sub.2O (compound isolated at 30% of CH.sub.3CN) to afford compound Int-60d (38.0 g, 84%) as a colourless gum: ESI (m/z) [C.sub.19H.sub.40N.sub.2O.sub.12+H].sup.+ 489.

[0638] Preparation of Int-60: To a stirred solution of Int-60d (38.0 g, 61.4 mmol) in MeOH (100 mL) was added HCl (3M in MeOH) (100 mL) and reaction stirred at ambient temperature for 16 h. Then the volatiles were removed under reduced pressure and lyophilized for 48 h to afford compound Int-60 (34.0 g, 94%) as an off-white gum; .sup.1H NMR (400 MHz, D.sub.2O) 4.22-4.18 (m, 1H), 4.10-4.05 (m, 1H), 3.81-3.58 (m, 10H), 3.57-3.35 (m, 8H); ESI (m/z) [C.sub.14H.sub.32N.sub.2O.sub.10+H].sup.+ 389; UPLC AUC 98.3% (R.sub.t=1.23 min); UPLC method AA Example Compounds Example 1

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(3-hydroxypropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-1)

##STR00370##

[0639] Preparation of Exp-1a: To a stirred solution of Int-16 (0.70 g, 1.67 mmol) in MeOH/H.sub.2O (30 mL; 2:1), was added Pd(OH).sub.2 (0.20 g, 10% wet by weight) at ambient temperature under nitrogen. The nitrogen atmosphere was replaced with H.sub.2 (1 atm) and reaction mixture was stirred for 16 h. The reaction mixture was filtered through a celite bed, the solids were washed with MeOH (25 mL) and the filtrate was concentrated to afford Exp-1a (0.55 g, crude) as a colorless gum. ESI (m/z) [C.sub.19H.sub.25NO+H].sup.+ 284.

[0640] Preparation of Exp-1: To a stirred solution of compound Exp-1a (0.15 g, 0.53 mmol) in DMF (2 mL) was added methyl (3,5-diamino-6-chloropyrazine-2-carbonyl)carbamimidothioate hydroiodide (compound 1, 0.220 g, 0.84 mmol) and DIPEA (0.75 mL, 4.24 mmol) at ambient temperature under nitrogen. The reaction mixture was warmed and stirred at 50 C. for 4 h. The resulting mixture was concentrated under reduced pressure and the crude was purified by reverse phase column chromatography using 0-45% CH.sub.3CN in H.sub.2O buffered with 0.1% TFA to afford compound Exp-1 (0.13 g, 30%) as trifluoroacetate salt as an off-white solid. .sup.1H NMR (400 MHz, CD.sub.3OD-d.sub.4) d 7.43-7.38 (m, 4H), 7.18-7.14 (m, 4H), 3.51 (t, J=16.0 Hz, 2H), 3.25-3.23 (m, 2H), 2.66-2.59 (m, 4H), 1.78-1.72 (m, 2H), 1.69-1.63 (m, 4H); ESI (m/z) [C.sub.25H.sub.30ClN.sub.7O.sub.2+H].sup.+ 497; HPLC AUC=87.4%, t.sub.R=5.97 min, Method C.

Example 2

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(2-hydroxyethoxy)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-2)

##STR00371##

[0641] Preparation of Exp-2a: To a stirred solution of compound Int-19 (1.20 g, 2.60 mmol) in IPA/H.sub.2O (7:3, 15 mL) was added to 20% Pd(OH).sub.2 on carbon 50% wet (0.15 g, 50% wt/wt) at room temperature under nitrogen atmosphere. The nitrogen atmosphere was replaced with H.sub.2 (balloon) and the reaction mixture was stirred for 6 h. The reaction mixture was filtered through celite and the solids were washed with IPA/H.sub.2O (60 mL). The combined filtrates were concentrated under reduced pressure to afford compound Exp-2a (0.65 g, 76%) as a colorless gum. ESI (m/z) [C.sub.20H.sub.25NO.sub.3+H].sup.+ 328.

[0642] Preparation of Exp-2b: To a stirred solution of compound Exp-2a (0.65 g, 1.98 mmol) in DMF (10 mL) was added compound 1 (0.51 g, 1.98 mmol) and DIPEA (2.0 mL, 11.92 mmol) at room temperature under nitrogen. The reaction mixture was stirred at 60 C. for 8 h. The reaction mixture was allowed to cool, then was concentrated under reduced pressure. The crude product was purified by preparative-HPLC (column; Gemini C18, 10 um, 30150 mm; mobile phase A: 0.05% TFA in water, B: CH.sub.3CN) to afford compound Exp-2b (0.35 g, 35%) as an off-white solid. ESI (m/z) [C.sub.26H.sub.30ClN.sub.7O.sub.4+H].sup.+ 540.

[0643] Preparation of Exp-2: To a solution of compound Exp-2b (0.3 g, 0.55 mmol) in anhydrous THF (15 mL) was added to 2.0 M LiBH.sub.4 (0.83 mL, 0.83 mmol) at 0 C. The reaction mixture was warmed to room temperature and stirred for 1.5 h. The reaction mixture was quenched with H.sub.2O at 0 C. The volatiles were removed under reduced pressure and the crude was purified by C-18 Reverse-Phase column chromatography using 25-35% CH.sub.3CN in H.sub.2O with 0.1% HCl buffer to afford compound Exp-2 (0.15 g, 54%) as an off-white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 11.5 (s, 1H), 9.26 (m, 1H), 8.94-8.75 (m, 2H), 7.60-7.50 (m, 4H), 8.43 (s, 2H), 7.29 (d, J=8.0 Hz, 2H), 7.01 (d, J=8.0 Hz, 2H), 4.03 (t, J=5.2 Hz, 2H), 3.72 (t, J=5.2 Hz, 2H), 3.35 (m, 2H), 2.64 (t, J=6.4 Hz, 2H), 1.72-1.53 (m, 4H); ESI (m/z) [C.sub.24H.sub.28ClN.sub.7O.sub.3+H].sup.+ 498; HPLC AUC=98.2%, t.sub.R=6.67 min, Method J.

Example 3

Preparation of 3,5-diamino-6-chloro-N(N-(4-(2-chloro-4-(3-hydroxypropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-14)

##STR00372##

[0644] Preparation of Exp-3a: To a stirred solution of Int-6c (3.73 g, 11.6 mmol) and compound Int-1 (2.00 g, 4.64 mmol) in a mixture of DMF/H.sub.2O (30 mL:6 mL) was added Na.sub.2CO.sub.3 (1.47 g, 13.90 mmol) at room temperature. The reaction mixture was degassed for 20 min with Argon, Pd(PPh.sub.3).sub.4 (313 mg, 0.278 mmol) was added, then warmed to 100 C. and stirred for 4 h. The reaction mixture was cooled to room temperature, water (50 mL) was added then extracted with EtOAc (250 mL). The combined organic layer was washed with cold water followed by brine solution (30 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The obtained crude material was purified by combi-flash column chromatography eluting with (1%-100% EtOAc in hexanes) to afford Exp-3a (1.10 g, 50%) as an off-white solid. ESI (m/z) [C.sub.28H.sub.28ClNO.sub.4+H].sup.+ 478.

[0645] Preparation of Exp-3b: A stirred solution of Exp-3a (0.60 g, 1.25 mmol) in IPA (15 mL) and CH.sub.3COOH (0.5 mL), was charged with PtO.sub.2 (100 mg). The reaction mixture was stirred under H.sub.2 balloon pressure for 16 h. The resulting reaction mixture was filtered through celite and the filtrate was again subjected to the same reaction (with fresh PtO.sub.2) for 16 h. Reaction mixture was filtered through celite and filtrate was concentrated under reduced pressure. The obtained crude material was purified combi-flash chromatography eluting with (1%-10% MeOH in dichloromethane) to afford Exp-3b (250 mg, 57%) as a light-yellow liquid. ESI (m/z) [C.sub.20H.sub.24ClNO.sub.2+H].sup.1346.

[0646] Preparation of Exp-3c: A stirred solution of compound Exp-3b (250 mg, 0.44 mmol) and compound 1 (169.9 mg, 0.44 mmol) in DMF (5.0 mL), was charged with DIPEA (0.24 mL, 1.34 mmol) at room temperature. The resultant reaction mixture was warmed to 60 C. and stirred for 16 h. The reaction mixture was poured into water (30 mL), the precipitated solids were filtered, and dried under reduced pressure to afford compound Exp-3c (350 mg, 83%) as a yellow solid. ESI (m/z) [C.sub.26H.sub.29Cl.sub.2N.sub.7O.sub.3+H].sup.+ 558.

[0647] Preparation of Exp-3: A stirred solution of compound Exp-3c (180 mg, 0.32 mmol) in THF (5.0 mL), was charged with 2M LiBH.sub.4 in THF (0.48 mL, 0.96 mmol) slowly dropwise at 0 C. under nitrogen, gradually allowed to warm to room temperature then stirred for 5 h. The reaction mixture was quenched with water and the pH of the solution was adjusted to 2 with 2N HCl. The precipitated solids were filtered, washed with 50% ACN in water and dried under reduced pressure to afford compound Exp-3 (60 mg, 32%) as a light-yellow solid. .sup.1H NMR (400 MHz, CD.sub.3OD) 7.24-7.08 (m, 7H), 3.49 (t, J=6.4 Hz, 2H), 2.27 (t, J=6.4 Hz, 2H), 2.67-2.59 (m, 4H), 1.79-1.65 (m, 6H); ESI (m/z) [C.sub.25H.sub.29Cl.sub.2N.sub.7O.sub.2+H].sup.+ 530; HPLC AUC=95.1%, t.sub.R=6.87 min, Method J.

Example 4

Preparation of 3,5-diamino-N(N-(4-(4-(3-aminopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide trifluoroacetate salt (Exp-4)

##STR00373##

[0648] Preparation of Exp-4a: To a stirred solution of Int-17c (0.50 g, 0.96 mmol) in EtOH (10 mL) was added 10% Pd on carbon (0.20 g, 40% wt) at room temperature under nitrogen atmosphere. The nitrogen atmosphere was replaced with a H.sub.2 filled balloon (1 atm) and reaction mixture was stirred for 16 h. The reaction mixture was filtered through celite bed and washed with MeOH. The filtrate was concentrated under reduced pressure to afford compound Exp-4a (0.32 g, 86%) as an off-white solid; ESI (m/z) [C.sub.24H.sub.34N.sub.2O.sub.2+H].sup.+ 383.

[0649] Preparation of Exp-4b: To a stirred solution of compound Exp-4a (0.29 g, 0.75 mmol) in EtOH (5 mL) was added compound 1 (0.51 g, 1.97 mmol) and diisopropylethylamine (0.78 g, 6.07 mmol) at room temperature under nitrogen atmosphere. The reaction mixture was stirred at 70 C. for 6 h. The resulting mixture was cooled to room temperature and concentrated under reduced pressure to get crude product which was purified by reverse phase (C-18) column chromatography using 0-30% CH.sub.3CN in H.sub.2O to afford compound Exp-4b (0.20 g, 64%) as an off-white solid. ESI (m/z) [C.sub.30H.sub.39ClN.sub.8O.sub.3+H].sup.+ 595.

[0650] Preparation of Exp-4: To a stirred solution of compound Exp-4b (0.15 g, 0.25 mmol) in dichloromethane (3 mL) was added TFA (0.50 mL, 3.3 wt/v) at 0 C. The reaction mixture was warmed to room temperature and stirred for 4 h. The reaction mixture was concentrated under reduced pressure and the crude was purified by reverse phase (C-18) column chromatography using 0-20% CH.sub.3CN in H.sub.2O buffer 0.1% TFA to afford Exp-4 (0.05 g, 35%) as a pale-yellow solid. .sup.1H NMR (400 MHz, MeOD) 7.45-7.41 (m, 4H), 7.19 (d, J=8.0 Hz, 4H), 3.27-3.24 (m, 2H), 2.88-2.84 (t, J=8.0 Hz, 2H), 2.67-2.62 (m, 4H), 1.93-1.85 (m, 2H), 1.67-1.64 (m, 4H); ESI (m/z) [C.sub.25H.sub.31ClN.sub.8O+H].sup.+ 495; HPLC AUC=96.5%, t.sub.R=11.72 min, Method C.

Example 5

Preparation of 3,5-diamino-N(N-(4-(4-(2-aminoethoxy)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-5)

##STR00374##

[0651] Preparation of Exp-5a: To a stirred solution of compound Int-19a (1.20 g, 3.20 mmol) and triphenylphosphine (1.25 g, 4.80 mmol) in THF (60 mL) was added to DIAD (0.97 g, 4.80 mmol) at 0 C. The reaction mixture was stirred for 5 min and a solution of N-Boc-ethanolamine (0.61 g, 3.84 mmol) in THF (15 mL) was added. The resulted mixture was warmed to 60 C. and stirring was continued for 8 h. The reaction mixture was diluted with H.sub.2O (100 mL) and extracted with EtOAc (2200 mL). The combined organic layer was washed with brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and evaporated under reduced pressure to obtain the crude product. The crude was purified by flash column chromatography (silica gel 120 g column) using 15-20% EtOAc in hexanes to afford compound Exp-5a (0.78 g, 47%) as an off-white solid. ESI (m/z) [C.sub.31H.sub.38N.sub.2O.sub.5+H].sup.+ 519.

[0652] Preparation of Exp-5b: To a stirred solution of compound Exp-5a (0.9 g, 1.73 mmol) in IPA/H.sub.2O (11.0 mL, 10:1) was added to 20% Pd(OH).sub.2 on carbon 50% wet (0.16 g, 25% wt. substrate) at room temperature under nitrogen atmosphere. The nitrogen atmosphere was replaced with H.sub.2 (balloon) and the reaction mixture was stirred for 6 h. The reaction mixture was filtered through celite pad, and the solids were washed with IPA and H.sub.2O (20 mL, 1:1). The filtrate was concentrated under reduced pressure to afford compound Exp-5b (0.55 g, 82%) as an off-white sticky solid. ESI (m/z) [C.sub.23H.sub.32N.sub.2O.sub.3+H].sup.+ 385.

[0653] Preparation of Exp-5c: To a stirred solution of compound Exp-5b (0.55 g, 1.43 mmol) in DMF (5.0 mL) was added to compound 1 (0.410 g, 1.57 mmol) and DIPEA (1.45 mL, 8.59 mmol) at room temperature under nitrogen atmosphere. The reaction mixture was stirred at 60 C. for 6 h. The reaction mixture was cooled then concentrated under reduced pressure. The obtained crude product was purified by C-18 Reverse-Phase column chromatography using 60-75% CH.sub.3CN in H.sub.2O to afford compound Exp-5c (0.3 g, 35%) as a light yellow solid. ESI (m/z) [C.sub.29H.sub.37ClN.sub.8O.sub.4+H].sup.+ 597.

[0654] Preparation of Exp-5: To a stirred solution of compound Exp-15c (0.3 g, 0.50 mmol) in 1,4-dioxane (3 mL) was added to a solution of 4 M HCl in 1,4-dioxane (0.5 mL, 5 wt/v) at 0 C. The reaction mixture was warmed to room temperature stirred for 2 h. The reaction mixture was concentrated, and the crude was co-distilled with 1,4-dioxane (5 mL) and MTBE (5 mL). The resulting solid was dried under reduced pressure to afford compound Exp-5 (0.11 g, 22%) as a light yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 10.52 (s, 1H), 9.26 (m, 1H), 8.94-8.75 (m, 2H), 8.58-8.51 (m, 4H), 8.45 (s, 2H), 7.28 (d, J=8.0 Hz, 2H), 7.03 (d, J=8.0 Hz, 2H), 4.01 (t, J=5.2 Hz, 2H), 3.74 (t, J=5.2 Hz, 2H), 3.33 (m, 2H), 2.65 (t, J=6.4 Hz, 2H), 1.71-1.555 (m, 4H); ESI (m/z) [C.sub.24H.sub.29ClN.sub.5O.sub.2+H].sup.+ 497; HPLC AUC=99.6%, t.sub.R=7.69 min, Method J.

Example 6

Preparation of 3,5-diamino-N(N-(4-(4-(3-aminopropyl)-2-chloro-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-6)

##STR00375##

[0655] Preparation of Exp-6a: A stirred solution of Int-1 (1.60 g, 3.71 mmol) in DMF/Water (20/6 mL) was degassed with argon for 30 minutes. Int-6 (2.58 g, 8.012 mmol), Na.sub.2CO.sub.3 (1.18 g, 11.13 mmol) and Pd(PPh.sub.3).sub.4 (257 mg, 0.22 mmol) were added, the reaction mixture was warmed to 100 C. and stirred for 6 h. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (3100 mL). The combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The obtained crude was purified by combiflash chromatography eluting with 40% EtOAc in hexanes to afford Exp-6a (0.850 g, 47%) as an off-white solid; ESI (m/z) [C.sub.28H.sub.30ClNO.sub.4+H].sup.+ 480. (Reduction under palladium coupling conditions has been observed; see D. P. Ojha, K. Gadde, K. R. Prabhu, Org. Lett., 2016, 18, 5062-5065.)

[0656] Preparation of Exp-6b: A stirred solution of Exp-6a (850 mg, 1.88 mmol) in THF (10 mL) was charged with 2M LiBH.sub.4 in THF (0.28 mL, 5.65 mmol) slowly dropwise at 0 C. under nitrogen, gradually allowed to room temperature then stirred for 16 h. The reaction mixture was quenched with water (30 mL) at 0 C., followed by 2 N HCl (10 mL) slowly at 0 C. then stirred for 15 minutes. The aqueous reaction mixture was extracted with EtOAc (2100 mL), the combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The crude material was purified by combi-flash column chromatography eluting with (10-100% EtOAc in hexanes) to affords Exp-6b (750 mg, 93%) as a white solid. ESI (m/z) [C.sub.27H.sub.30ClNO.sub.3+H].sup.+ 452

[0657] Preparation of Exp-6c: A stirred solution of Exp-6b (750 mg, 1.66 mmol) in dichloromethane (15 mL), was charged with Et.sub.3N (0.4 mL, 1.66 mmol) followed by MsCl (190 mg, 1.66 mmol) at 0 C. The reaction mixture was warmed to room temperature and stirred for 4 h. Reaction mixture was diluted with dichloromethane (50 mL), washed with water (20 mL) then brine solution (20 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure to afford crude which was washed with MTBE (210 mL) to affords Exp-6c (900 mg, crude) as a brown solid which was used in the next step without purification. ESI (m/z) [C.sub.28H.sub.32ClNO.sub.5S+H].sup.+ 530.

[0658] Preparation of Exp-6d: A stirred solution of Exp-6c (1.1 g, 2.07 mmol) in DMF (10.0 mL), was charged with NaN.sub.3 (135 mg, 2.07 mmol) at room temperature then stirred for 5 h. The reaction mixture was diluted with water then extracted with EtOAc (3100 mL). The combined organic layer was washed with brine solution (50 mL), dried over anhydrous NaSO.sub.4, filtered, and concentrated under reduced pressure to afford compound Exp-6d (800 mg, crude) as a brown color gummy solid. ESI (m/z) [C.sub.27H.sub.29ClN.sub.4O.sub.2+H].sup.+ 477.

[0659] Preparation of Exp-6e: A stirred solution of Exp-6d (800 mg, 1.67 mmol) in THF (15.0 mL) and H.sub.2O (5.0 mL), was charged with PPh.sub.3 (440 mg, 1.67 mmol) at room temperature then stirred for 12 h. The reaction mixture was diluted with water (20 mL) then extracted into dichloromethane (330 mL). The organic layer was concentrated under reduced pressure and the resulting Exp-6e (1.60 g, crude) was used in the next step without purification. ESI (m/z) [C.sub.27H.sub.31ClN.sub.2O.sub.2+H].sup.+ 451.

[0660] Preparation of Exp-6f: A stirred solution of Exp-6e (1.60 g, 3.55 mmol) in THF (10.0 mL) was added (Boc).sub.2O (780 mg, 3.5 mmol) then stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure and the resulting crude was purified by combi-flash column chromatography eluting with (50% EtOAc in hexanes) to afford Exp-6f (450 mg, 49% for 3 steps) as an off-white solid. ESI (m/z) [C.sub.32H.sub.39ClN.sub.2O.sub.4+H].sup.+ 551.

[0661] Preparation of Exp-6g: A stirred solution of Exp-6f (145 mg, 0.26 mmol) in water (7.50 mL) and CH.sub.3CO.sub.2H (2.50 mL), EtOAc (1.0 mL) was charged with PtO.sub.2 (30 mg) then stirred under H.sub.2 (balloon) for 12 h. Reaction mixture was filtered through celite, and the filtrate was concentrated under reduced pressure. The obtained crude material was purified by reverse phase column chromatography using 40% acetonitrile in water to afford Exp-6g (120 mg, 95%) as off-white gummy solid ESI (m/z) [C.sub.24H.sub.33ClN.sub.2O.sub.2+H].sup.+ 417.

[0662] Preparation of Exp-6h: A stirred solution of Exp-6g (125 mg, 0.30 mmol) and compound 1 (114 mg, 0.30 mmol) in DMF (10.0 mL) was charged with DIPEA (0.26 mL, 1.80 mmol) at room temperature. The reaction mixture was warmed to 60 C. and stirred for 6 h. The reaction mixture was poured in beaker contains water (30 mL), the precipitated solid was filtered and dried under reduced pressure. The residue was purified by C-18 reverse phase column chromatography (40% CH.sub.3CN/0.01% AcOH in H.sub.2O) to afford compound Exp-6h (70 mg, 42%) as a yellow solid. ESI (m/z) [C.sub.30H.sub.38Cl.sub.2N.sub.8O.sub.3+H].sup.+ 629.

[0663] Preparation of Exp-6: A stirred solution of compound Exp-6h (90 mg, 0.14 mmol) in dichloromethane (5.0 mL), was charged with 4M HCl in 1,4-dioxane (10 mL) at room temperature then stirred for 6 h. The reaction mixture concentrated under reduced pressure, the residue was purified by C-18 reverse phase column chromatography (40% CH.sub.3CN in H.sub.2O) to afford compound Exp-6 (60 mg, 70%) as a yellow solid. .sup.1H NMR (400 MHz, CD.sub.3OD-d.sub.6) 7.31-7.28 (s, 1H), 7.25-7.17 (m, 5H), 7.15-7.12 (m, 1H), 3.29-3.26 (t, J=4.4 Hz, 2H), 2.90-2.86 (t, J=8.0 Hz, 2H), 2.68-2.63 (m, 4H), 1.95-1.85 (m, 2H), 1.75-1.62 (m, 4H), ESI (m/z)) [C.sub.25H.sub.30Cl.sub.2N.sub.8OH].sup. 527; HPLC AUC=93.8%, t.sub.R=6.43 min, Method J.

Example 7

Preparation of 4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-carboxylic acid (Exp-7)

##STR00376##

[0664] Preparation of Exp-7a: A suspension of Int-8 (350 mg, 0.838 mmol) in EtOH (10 mL), water (2.0 mL) and AcOH (0.5 mL) was heated to 50 C. and stirred for 30 min. After a clear brown solution observed, 10% Pd/C (50 mg) was added, and the resultant suspension was stirred at 50 C. under hydrogen (balloon) for 4 h. After filtration, the solvent was removed, to afford compound Exp-7a (264 mg, 92%) as colorless oil: .sup.1H NMR (500 MHz, CD.sub.3OD) 8.07 (d, J=8.5 Hz, 2H), 7.71 (d, J=8.5 Hz, 2H), 7.61 (d, J=8.5 Hz, 2H), 7.33 (d, J=8.5 Hz, 2H), 3.92 (s, 3H), 2.94 (t, J=7.0 Hz, 2H), 2.74 (t, J=7.0 Hz, 2H), 1.76-1.69 (m, 4H); ESI (m/z) [C.sub.18H.sub.21N.sub.9O.sub.2+H].sup.+ 284.

[0665] Preparation of Exp-7b: To a solution of Exp-7a (50.0 mg, 0.146 mmol) in DMF (2.0 mL) and EtOH (1.0 mL) was added compound 1 (56.6 mg, 0.146 mmol) and DIPEA (113 mg, 0.874 mmol) at room temperature. The resultant mixture was stirred at 65 C. for 6 h. After concentration, water (6.0 mL) was added. The resulting yellow solid was collected, washed with water (3.0 mL) and dried, to afford compound Exp-7b (65 mg, 90%) as a yellow solid: ESI (m/z) [C.sub.24H.sub.26ClN.sub.7O.sub.3+H].sup.+ 496.

[0666] Preparation of Exp-7: To a solution of Exp-7b (280 mg, 0.565 mmol) in dioxane (50 mL) and water (15 mL) was added LiOH monohydrate (237 m, 56.5 mmol). The resulting solution was stirred at room temperature for 16 h. After the pH value was adjusted to 3 with 4 N HCl, the solvent was removed. Water (20 mL) was added, and the yellow solid was filtered out. The semi dry solid was re-suspended in MeOH (10 mL) and EtOH (10 mL) and stirred at 40 C. for 30 min. After cooling, the yellow solid was collected by filtration and dried to afford Exp-7 (188 mg, 64%) as a yellow solid: .sup.1H NMR (500 MHz, DMSO-d.sub.6) 12.93 (br s, 1H), 10.45 (br s, 1H), 9.18 (br s, 1H), 8.90 (br s, 1H), 8.73 (br s, 1H), 7.99 (d, J=8.5 Hz, 2H), 7.77 (d, J=8.5 Hz, 2H), 7.67 (d, J=8.0 Hz, 2H), 7.42 (br s, 2H), 7.35 (d, J=8.0 Hz, 2H), 3.33-3.30 (m, 2H), 2.68 (t, J=6.0 Hz, 2H), 1.68-1.59 (m, 4H); ESI (m/z) [C.sub.23H.sub.24ClN.sub.7O.sub.3+H].sup.+ 482; HPLC AUC=99.3%, t.sub.R=9.40 min, Method A.

Example 8

Preparation of 3-(3-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoic acid (Exp-8)

##STR00377##

[0667] Preparation of Exp-8a: To a solution of Int-52 (400 mg, 0.97 mmol) in CH.sub.2Cl.sub.2 (10 mL), was added 4M HCl in 1,4-dioxane (5.0 mL) followed by stirring at room temperature for 3 h. The reaction mixture concentrated under reduced pressure to complete dryness. The obtained gummy solid was washed with MTBE (20 mL), filtered and dried under reduced pressure to afford Exp-8a (320 mg, 94%) as light brown solid: ESI (m/z) [C.sub.20H.sub.25NO.sub.2+H].sup.+ 312.

[0668] Preparation of Exp-8b: To a solution of Exp-8a (320 mg, 0.92 mmol) and compound 1 (393 mg, 1.01 mmol) in DMF (5.0 mL), was added DIPEA (0.45 mL, 2.76 mmol). The reaction mixture was heated to 60 C. then stirred for 16 h. The reaction mixture was concentrated under reduced pressure, then water (50 mL) was added followed by stirring for 30 min. The resulting solids were filtered, washed with water (20 mL) and dried under reduced pressure to afford Exp-8b (350 mg, 72%) as brown solid: ESI (m/z) [C.sub.26H.sub.30ClN.sub.7O.sub.3+H].sup.+ 525.

[0669] Preparation of Exp-8: To a solution of Exp-8b (350 mg, 0.667 mmol) in a mixture of THF (10 mL), methanol (4.0 mL) and water (4.0 mL), was added LiOH.Math.H.sub.2O (56 mg, 1.33 mmol) followed by stirring at room temperature for 5 h. The reaction mixture was diluted with water (5.0 mL) and acidified to pH=2 with 2N aqueous HCl solution. The resulting solids were filtered and washed with water (20 mL). The crude was purified by reverse phase combi-flash chromatography eluted with 10-100% ACN in water (compound isolated at 45% of ACN) to afford Exp-8 (120 mg, 51%) as HCl salt, yellow in color: .sup.1H NMR [(400 MHz, DMSO-d.sub.6 (D.sub.2O exchange)] 7.52 (d, J=8 Hz 2H), 7.44 (s, 2H), 7.39-7.35 (m, 1H), 7.29 (d, J=8.0 Hz, 2H), 7.19 (d, J=7.6 Hz, 1H), 3.28-3.20 (m, 2H), 2.84-2.80 (m, 2H), 2.73-2.67 (m, 2H), 2.05-2.41 (m, 2H), 1.70-1.60 (m, 4H); ESI (m/z) [C.sub.25H.sub.28ClN.sub.7O.sub.3+H].sup.+ 510; HPLC AUC=98.8%, t.sub.R=6.85 min, Method I.

Examples 9-17

[0670] Preparation of Examples 9-17: In a scheme analogous to the one used for preparation of Exp-8, Examples 9-17 were synthesized from the corresponding intermediates. Intermediates with unsaturation in the side chains were reduced during the removal of the Cbz protecting group:

TABLE-US-00032 Exp-# Int-# Structure Spectra 9 9 [00378] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.34-7.26 (m, 4H), 3.59 (s, 2H), 3.30-3.25 (m, 2H), 2.73 (t, J = 7.0 Hz, 2H), 1.81-1.71 (m, 4H); ESI (m/z) [C.sub.24H.sub.26ClN.sub.7O.sub.3 + H].sup.+ 496; HPLC AUC = 94.0%, t.sub.R = 5.15 min, Method A. 10 10 [00379] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.55-7.53 (m, 4H), 7.30-7.28 (m, 4H), 6.70-6.61 (m, 2H), 3.23-3.17 (m, 2H), 2.86-2.82 (m, 2H), 2.69-2.61 (m, 4H), 2.56-2.53 (m, 4H), 2.35-2.32 (m, 1H), 1.65-1.63 (m, 2H), 1.56-1.54 (m, 2H); ESI (m/z) [C.sub.25H.sub.28ClN.sub.7O.sub.3 + H].sup.+ 511; HPLC AUC = 100%, t.sub.R = 10.24 min, Method C 11 11 [00380] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.28-7.23 (m, 4H), 3.45-3.35 (m, 2H), 2.74- 2.66 (m, 4H), 2.31 (t, J = 7.0 Hz, 2H), 1.96-1.90 (m, 2H), 1.81- 1.71 (m, 4H); ESI (m/z) [C.sub.26H.sub.30ClN.sub.7O.sub.3 + H].sup.+ 524; HPLC AUC = 95.1%, t.sub.R = 11.52 min, Method A 12 12 [00381] .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.53-7.50 (m, 2H), 7.44- 7.42 (m, 2H), 7.40-7.37 (m, 3H), 7.19-7.16 (m, 1H), 3.36-3.33 (m, 2H), 3.30-3.29 (m, 2H), 2.98-2.94 (m, 2H), 2.75-2.61 (m, 2H), 1.80-1.71 (m, 4H); ESI (m/z) [C.sub.25H.sub.28ClN.sub.7O.sub.3 + H].sup.+ 510; HPLC AUC = 97.2%, t.sub.R = 6.85 min, Method I 13 13 [00382] .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.16 (d, J = 8.0 Hz, 2H), 7.10 (d, J = 16 Hz, 2H), 7.05 (s, 1H), 6.91 (d, 2H), 3.26 (d, J = 6.4 Hz, 2H), 2.75 (d, J = 7.6 Hz, 2H), 2.63 (d, J = 6.8 Hz, 2H), 2.52 (d, J = 7.6 Hz, 2H), 2.90 (S, 3H), 1.71-1.62 (m, 4H), ESI (m/z) [C.sub.26H.sub.30ClN.sub.7O.sub.3 + H].sup.+ 524; HPLC AUC = 98.2%, t.sub.R = 6.98 min, Method J 14 14 [00383] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.49 (d, J = 8.5 Hz, 2H), 7.36 (d, J = 8.5 Hz, 2H), 7.28 (d, J = 8.5 Hz, 2H), 7.20 (d, J = 8.5 Hz, 2H), 3.39 (t, J = 6.0 Hz, 2H), 2.92-2.89 (m, 2H), 2.81 (t, J = 7.0 Hz, 2H), 2.49- 2.45 (m, 2H), 2.09-2.06 (m, 2H); ESI (m/z) [C.sub.24H.sub.26ClN.sub.7O.sub.3 + H].sup.+ 496; UPLC AUC = 94.2%, t.sub.R = 3.28 min, Method B 15 15 [00384] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.47-7.41 (m, 4H), 7.24-7.22 (m, 4H), 3.33-3.30 (m, 2H), 2.93 (t, J = 9.0 Hz, 2H), 2.70 (t, J = 8.0 Hz, 2H), 2.62-2.59 (m, 2H), 1.76-1.69 (m, 4H), 1.44- 1.41 (m, 2H); ESI (m/z) [C.sub.26H.sub.30ClN.sub.7O.sub.3 + H].sup.+ 524; UPLC AUC = 95.3%, t.sub.R = 3.63 min, Method B 16 19 [00385] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.25 (S, 1H), 8.92-8.70 (m, 2H), 7.60-7.50 (m, 4H), 8.43 (s, 2H), 7.27 (d, J = 8.0 Hz, 2H), 6.95 (d, J = 8.0 Hz, 2H), 4.80 (s, 2H), 3.35 (m, 2H), 2.65 (t, J = 6.8 Hz, 2H), 1.72-1.50 (m, 4H); ESI (m/z) [C.sub.24H.sub.26ClN.sub.7O.sub.4 + H].sup.+ 512; HPLC AUC = 99.1%, t.sub.R = 6.69 min, Method J 17 Exp- 3b [00386] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.26-7.12 (m, 7H), 3.28-3.26 (m, 2H), 2.84 (t, J = 7.6 Hz, 2H), 2.65 (t, J = 7.2 Hz, 2H), 2.54 (t, J = 7.6 Hz, 2H), 1.73- 1.62 (m, 4H); ESI (m/z) [C.sub.25H.sub.27Cl.sub.2N.sub.7O.sub.3 + H].sup.+ 544; HPLC AUC = 91.1%, t.sub.R = 6.85 min, Method K

Example 18

Preparation of methyl (3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysinate (Exp-18)

##STR00387##

[0671] Preparation of Exp-18a: To a solution of Int-36 (89.0 mg, 0.165 mmol) in EtOH (1.0 mL) and DMF (1.0 mL) was added compound 1 (70.5 mg, 0.181 mmol) at room temperature. The resultant mixture was stirred at 70 C. for 8 h. After solvent removal, water (3.0 mL) was added. The resulting solids were precipitated, filtered and dried, to afford Exp-18a (112 mg, 90%) as a yellow solid: ESI (m/z) [C.sub.37H.sub.50ClN.sub.9O.sub.6+H].sup.+ 752.

[0672] Preparation of Exp-18: To a solution of Exp-18a (312 mg, 0.415 mmol) in DCM (12 mL) was added TFA (3.0 mL) at room temperature. The resultant mixture was stirred at room temperature for 2 h. After the solvent was removed, the residue was dissolved in MeOH (12 mL) and 3 N HCl in MeOH (3.0 mL) was added. The solution was concentrated then azeotroped with MeOH (10 mL2). The resulting residue was purified by reverse phase column (5%90% CH3CN in water, product came out at 30%), to afford Exp-18 (272 mg, 90%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.29-7.28 (m, 4H), 4.45-4.42 (m, 1H), 3.70 (s, 3H), 3.45-3.37 (m, 2H), 2.98-2.85 (m, 4H), 2.73 (t, J=7.0 Hz, 2H), 2.59 (t, J=7.5 Hz, 2H), 1.82-1.64 (m, 8H), 1.42-1.38 (m, 2H); ESI (m/z) [C.sub.32H.sub.42ClN.sub.9O.sub.4+H].sup.+ 652; HPLC AUC=98.8%; t.sub.R=9.40 min, Method A.

Example 19

Preparation of (3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysine (Exp-19)

##STR00388##

[0673] Preparation of Exp-19a: To a solution of Exp-18a (112 mg, 0.149 mmol) in THF (1.5 mL), MeOH (1.5 mL) and water (0.5 mL) was added NaOH (59.5 mg, 1.489 mmol) at room temperature. The resultant mixture was stirred at room temperature for 3 h. After the solvent was removed, water (4.0 mL) was added. The pH was adjusted to 4 with 1 N HCl, and solid precipitation was observed. The solid was collected by filtration, washed with water and dried in oven, to afford Exp-19a (108 mg, 98%) as a yellow solid: ESI (m/z) [C.sub.36H.sub.48ClN.sub.9O.sub.6+H].sup.+ 738.

[0674] Preparation of Exp-19: To a solution of Exp-19a (82 mg, 0.111 mmol) in dioxane (2.0 mL) was added 4 N HCl in dioxane (2.0 mL) at room temperature. The resultant mixture was stirred at room temperature for 3 h. After the solvent was removed, the residue was purified by reverse phase column (10% to 90% of CH.sub.3CN in H.sub.2O, product came out at 30% to 90%), to afford Exp-19 (50 mg, 63%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.51-7.47 (m, 4H), 7.28-7.26 (m, 4H), 4.26-4.23 (m, 1H), 3.34-3.33 (m, 2H), 2.96-2.94 (m, 2H), 2.83 (t, J=7.5 Hz, 2H), 2.72 (t, J=7.0 Hz, 2H), 2.57 (t, J=7.5 Hz, 2H), 1.79-1.59 (m, 7H), 1.41-1.28 (m, 3H); ESI (m/z) [C.sub.31H.sub.40ClN.sub.9O.sub.4+H].sup.+ 638. HPLC AUC=100%; t.sub.R=9.08 min, Method A.

Example 20

Preparation of methyl N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.2.-methyl-L-lysinate (Exp-20)

##STR00389##

[0675] Preparation of Exp-20a: To a solution of Int-23 (300 mg, 0.695 mmol) in DMF (3.0 mL) was added HATU (291 mg, 0.765 mmol) followed by compound Int-46 (200 mg, 0.730 mmol) and DIPEA (270 mg, 2.086 mmol). The reaction mixture was stirred at rt for 1 h. The reaction mixture was partitioned with water (30 mL) and EtOAc (30 mL). The water phase was extracted with EtOAc (30 mL), and the combined EtOAc layers were concentrated, to afford compound Exp-20a (452 mg, 95%) as white solid: ESI (m/z) [C.sub.40H.sub.53N.sub.3O.sub.7+H].sup.+ 688.

[0676] Preparation of Exp-20b: To a solution of Exp-20a (420 mg, 0.611 mmol) in IPA (10 mL) was added Pd(OH).sub.2/C (50 mg). The reaction mixture was stirred under H.sub.2 (balloon) at rt for 2 h, then 35 C. for 2 h. After filtration, the solvent was removed, to afford Exp-20b (312 mg, 92%) as a white solid: ESI (m/z) [C.sub.32H.sub.47N.sub.3O.sub.5+H].sup.+ 554.

[0677] Preparation of Exp-20c: To a solution of Exp-20b (50 mg, 0.090 mmol) in DMF (1.0 mL) was added 1 (35.1 mg, 0.090 mmol) followed by DIPEA (46.7 mg, 0.361 mmol). The resultant solution was heated at 55 C. and stirred for 16 h. After the solvent was removed, the residue was purified by reverse phase column (10% to 60% of CH.sub.3CN in water, product came out at 40%), to afford Exp-20c (56.0 mg, 81%) as a yellow solid: ESI (m/z) [C.sub.38H.sub.52ClN.sub.9O.sub.6+H].sup.+ 766.

[0678] Preparation of Exp-20: Compound Exp-20c (285 mg, 0.372 mmol) was dissolved in 3 N HCl in MeOH (9.0 mL). The resultant solution was stirred at rt for 3 h, then at 35 C. for 1 h. The solvent was removed, and the residue was washed with CH.sub.3CN, to afford compound Exp-20 (248 mg, 90%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.50 (m, 4H), 7.31-7.27 (m, 4H), 5.04-5.01 (m, 1H), 3.71-3.69 (m, 3H), 3.45-3.33 (m, 2H), 2.97-2.71 (m, 11H), 2.02-2.00 (m, 1H), 1.81-1.65 (m, 7H), 1.40-1.31 (m, 2H); ESI (m/z) [C.sub.33H.sub.44ClN.sub.9O.sub.4+H].sup.+ 666; HPLC AUC=95.8%; t.sub.R=9.79 min, Method A.

Example 21

Preparation of N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.2.-methyl-L-lysine (Exp-21)

##STR00390##

[0679] Preparation of Exp-21a: To a stirred solution of Exp-20c (500 mg, 0.65 mmol) in a mixture of THF:MeOH:H.sub.2O (2:2:1, 10 mL), LiOH.Math.H.sub.2O (41 mg, 0.98 mmol) was added at 0 C. followed by stirring for 2 h at room temperature. The reaction mixture was dried completely under vacuum, cold water (20 mL) was added and the pH was adjusted to 4 with 2N HCl. The resulting solids were filtered, washed with water (20 mL) then dried under reduced pressure to afford Exp-21a (450 mg, 88%) as a white solid. ESI (m/z) [C.sub.37H.sub.50ClN.sub.9O.sub.6+H].sup.+ 752.

[0680] Preparation of Exp-21: To a stirred solution of Exp-21a (450 mg, 0.59 mmol) in 1,4-dioxane was added a solution 4.0 M HCl in dioxane (5 mL) at 0 C. The reaction mixture was stirred under nitrogen atmosphere at 0 C. for 2 h. The volatiles were removed under reduced pressure to afford Exp-21 (350 mg, 87%) as light yellow solid. .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.45-7.37 (mi, 4H), 7.23-7.14 (i, 4H), 5.02-4.95 (i, 1H), 3.30-3.23 (i, 2H), 2.90-2.84 (i, 4H), 2.83-2.76 (m, 2H), 2.74-2.66 (m, 2H), 2.65-2.59 (i, 2H), 2.01-1.85 (i, 1H), 1.78-1.53 (i, 8H), 1.31-1.17 (i, 2H); ESI (m/z) [C.sub.32H.sub.42ClN.sub.9O.sub.4+H].sup.+ 3652; HPLC AUC=96.4%; t.sub.R=6.34 min, Method O.

Examples 22-35

[0681] Preparation of Examples 22-35: In a scheme analogous to the one used for preparation of Examples-18-21, single amino acids were coupled to Int-23 and converted to corresponding Examples 22-35:

TABLE-US-00033 Exp-# AA Structure Spectra 22 D- Lys(Boc)- OCH.sub.3 [00391] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.28-7.27 (m, 4H), 4.44-4.41 (m, 1H), 3.69 (s, 3H), 3.44-3.43 (m, 2H), 2.97-2.85 (m, 4H), 2.73 (t, J = 7.5 Hz, 2H), 2.58 (t, J = 7.5 Hz, 2H), 1.85- 1.62 (m, 8H), 1.41- 1.36 (m, 2H); ESI (m/z) [C.sub.32H.sub.42ClN.sub.9O.sub.4 + H].sup.+ 652; HPLC AUC = 94.3%; t.sub.R = 9.21 min, Method A. 23 D- Lys(Boc)- OCH.sub.3 [00392] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.29-7.27 (m, 4H), 4.43-4.40 (m, 1H), 3.37- 3.36 (m, 2H), 2.96-2.86 (m, 4H), 2.73 (t, J = 7.0 Hz, 2H), 2.59 (t, J = 7.5 Hz, 2H), 1.89-1.63 (m, 8H), 1.42-1.40 (m, 2H); ESI (m/z) [C.sub.31H.sub.40ClN.sub.9O.sub.4 + H].sup.+ 638; HPLC AUC = 99.9%; t.sub.R = 6.34 min, Method I 24 Int-41 [00393] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.28-7.27 (m, 4H), 4.46-4.43 (m, 1H), 3.69 (s, 3H), 3.35-3.34 (m, 2H), 3.18-3.14 (m, 2H), 2.96-2.93 (m, 2H), 2.73 (t, J = 7.5 Hz, 2H), 2.58 (t, J = 7.5 Hz, 2H), 1.88- 1.55 (m, 8H); ESI (m/z) [C.sub.32H.sub.42ClN.sub.11O.sub.4 + H].sup.+ 680; HPLC AUC = 100%; t.sub.R = 9.38 min, Method A. 25 Int-41 [00394] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.51-7.47 (m, 4H), 7.28- 7.26 (m, 4H), 4.28-4.25 (m, 1H), 3.35-3.30 (m, 2H), 3.17- 3.13 (m, 2H), 2.98-2.93 (m, 2H), 2.72 (t, J = 7.0 Hz, 2H), 2.58 (t, J = 7.5 Hz, 2H), 1.82- 1.50 (m, 8H); ESI (m/z) [C.sub.31H.sub.40ClN.sub.11O.sub.4 + H].sup.+ 666; HPLC AUC = 100%; t.sub.R = 9.17 min, Method A. 26 D-(CH.sub.3) Lys(Boc)- OCH.sub.3 [00395] .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.43-7.40 (m, 4H), 7.22-7.178 (m, 4H), 4.97-4.88 (m, 1H), 3.61 (d, J = 8.08 Hz, 3H), 3.28- 3.22 (m, 3H), 2.87 (s, 3H), 2.86-2.83 (m, 1H), 2.83-2.76 (m, 2H), 2.71-2.67 (m, 2H), 2.66-2.60 (m, 2H), 1.96-1.84 (m, 1H), 1.78-1.52 (m, 7H), 1.31-1.12 (m, 2H); ESI (m/z) [C.sub.33H.sub.44ClN.sub.9O.sub.4 + H].sup.+ 666; HPLC AUC = 99.4%; t.sub.R = 8.50 min, Method W. 27 D-(CH.sub.3) Lys(Boc)- OCH.sub.3 [00396] .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.43-7.40 (m, 4H), 7.22-7.178 (m, 4H), 5.03-4.97 (m, 1H), 3.26 (t, J = 6.38, 2H), 2.88 (s, 3H), 2.86-2.77 (m, 3H), 2.73-2.61 (m, 5H), 1.98-1.88 (m, 1H), 1.77-1.52 (m, 7H), 1.30- 1.18 (m, 2H); ESI (m/z) [C.sub.32H.sub.42ClN.sub.9O.sub.4 + H].sup.+ 652; HPLC AUC = 98.5%; t.sub.R = 6.40 min, Method I. 28 Phe- OCH.sub.3 [00397] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.41-7.35 (m, 4H), 7.23-7.00 (m, 9H), 4.57-4.54 (m, 1H), 3.56 (s, 3H), 3.26-3.20 (m, 2H), 3.02-2.97 (m, 1H), 2.84- 2.61 (m, 3H), 2.63 (t, J = 7.2 Hz, 2H), 2.39 (t, J = 7.6 Hz, 2H), 1.72-1.61 (m, 4H); ESI (m/z) [C.sub.35H.sub.39ClN.sub.8O.sub.4 + H].sup.+ 671; HPLC AUC 98.2 % (R.sub.t = 9.54 min); HPLC AUC = 98.2%; t.sub.R = 9.54 min, Method S. 29 Phe- OCH.sub.3 [00398] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.41-7.34 (m, 4H), 7.18-7.02 (m, 9H), 4.55 (dd, J = 8.8, 4.8 Hz, 1H), 3.25 (t, J = 6.4 Hz, 2H), 3.16-3.03 (m, 1H), 2.86-2.80 (m, 1H), 2.75 (t, J = 7.6 Hz, 2H), 2.63 (t, J = 7.2 Hz, 2H), 2.39 (t, J = 7.6 Hz, 2H), 1.71-1.61 (m, 4H); ESI (m/z) [C.sub.34H.sub.37ClN.sub.8O.sub.4 + H].sup.+ 657; HPLC AUC 97.8% (R.sub.t = 9.13 min); HPLC AUC = 97.8%; t.sub.R = 9.13 min, Method T.. 30 Val- OCH.sub.3 [00399] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.39 (t, J = 8.0 Hz, 4H), 7.18 (dd, J = 8.4, 2.0 Hz, 4H), 4.19 (d, J = 6.0 Hz, 1H), 3.58 (s, 3H), 3.25 (t, J = 7.2 Hz, 2H), 2.85 (t, J = 7.6 Hz, 2H), 2.63 (t, J = 7.2 Hz, 2H), 2.53-2.48 (m, 2H), 1.98- 1.93 (m, 1H), 1.71-1.63 (m, 4H), 0.76-0.74 (m, 6H); ESI (m/z) [C.sub.31H.sub.39ClN.sub.8O.sub.4 + H].sup.+ 623; HPLC AUC = 99.3%; t.sub.R = 9.25 min, Method T. 31 Val- OCH.sub.3 [00400] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.41-7.36 (m, 4H), 7.19-7.16 (m, 4H), 4.17 (d, J = 5.2 Hz, 1H), 3.25 (t, J = 6.8 Hz, 2H), 2.86 (t, J = 7.6 Hz, 2H), 2.63 (t, J = 7.6 Hz, 2H), 2.55-2.47 (m, 2H), 2.04- 1.95 (m, 1H), 1.73-1.62 (m, 4H), 0.75-0.73 (m, 6H); ESI (m/z) [C.sub.30H.sub.37ClN.sub.8O.sub.4 + H].sup.+ 609; HPLC AUC = 99.0%; t.sub.R = 8.71 min, Method T. 32 Trp- OCH.sub.3 [00401] .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.48 (d, J = 8.23 Hz, 3H), 7.41 (d, J = 8.18 Hz, 2H), 7.36-7.24 (m, 3H), 7.16 (d, J = 8.20 Hz, 2H), 7.10-7.05 (m, 1H), 7.02-6.96 (m, 2H), 4.75- 4.71 (m, 1H), 3.63 (s, 3H), 3.37-3.32 (m, 2H), 3.27- 3.21 (m, 1H), 3.16-3.08 (m, 1H), 2.84 (t, J = 7.57 Hz, 2H), 2.72 (t, J = 6.98 Hz, 2H), 2.49 (t, J = 7.62 Hz, 2H), 1.84-1.67 (m, 4H); ESI (m/z) [C.sub.37H.sub.40ClN.sub.9O.sub.4 + H].sup.+ 710; HPLC AUC = 97.6%; t.sub.R = 9.25 min, Method T. 33 Trp- OCH.sub.3 [00402] .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.49 (d, J = 7.93 Hz, 1H), 7.43 (d, J = 8.09 Hz, 2H), 7.35 (d, J = 8.16 Hz, 2H), 7.27-7.21 (m, 3H), 7.10 (d, J = 8.21 Hz, 2H), 7.05-7.01 (m, 1H), 6.96-6.93 (m, 2H), 4.72- 4.69 (m, 1H), 3.31-3.27 (m, 3H), 3.13-3.07 (m, 1H), 2.78 (t, J = 7.53 Hz, 2H), 2.68 (t, J = 6.97 Hz, 2H), 2.44 (t, J = 7.73 Hz, 2H), 1.77-1.66 (m, 4H); ESI (m/z) [C.sub.36H.sub.38ClN.sub.9O.sub.4 + H].sup.+ 696; HPLC AUC = 99.2%; t.sub.R = 9.41 min, Method V. 34 Tyr- OCH.sub.3 [00403] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.50 (d, J = 8.0 Hz, 2H), 7.47 (d, J = 8.0 Hz, 2H), 7.27 (d, J = 8.0 Hz, 2H), 7.20 (d, J = 8.5 Hz, 2H), 6.93 (d, J = 8.5 Hz, 2H), 6.66 (d, J = 8.5 Hz, 2H), 4.61-4.58 (m, 1H), 3.65 (s, 3H), 3.36- 3.20 (m, 2H), 3.01-2.97 (m, 1H), 2.88-2.80 (m, 3H), 2.72 (t, J = 7.5 Hz, 2H), 2.51-2.48 (m, 2H), 1.81-1.71 (m, 4H); ESI (m/z) [C.sub.35H.sub.39ClN.sub.8O.sub.5 + H].sup.+ 687; UPLC AUC = 96.1%; t.sub.R = 3.94 min, Method B. 35 Tyr- OCH.sub.3 [00404] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.50 (d, J = 8.0 Hz, 2H), 7.45 (d, J = 8.0 Hz, 2H), 7.26 (d, J = 8.0 Hz, 2H), 7.20 (d, J = 8.5 Hz, 2H), 6.92 (d, J = 8.5 Hz, 2H), 6.61 (d, J = 8.5 Hz, 2H), 4.52-4.50 (m, 1H), 3.44-3.38 (m, 2H), 3.05-3.01 (m, 1H), 2.89- 2.84 (m, 3H), 2.72 (t, J = 7.5 Hz, 2H), 2.50-2.46 (m, 2H), 1.81-1.70 (m, 4H); ESI (m/z) [C.sub.34H.sub.37ClN.sub.8O.sub.5 + H].sup.+ 673; UPLC AUC = 96.8%; t.sub.R = 3.68 min, Method B.

Example 36

Preparation of methyl (S)-2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)-3-(pyridin-4-yl)propanoate (Exp 36)

##STR00405##

[0682] Preparation of 36a: To a solution of (S)-3-amino-2-oxo-4-(pyridin-4-yl)butanoic acid (3.00 g, 15.45 mmol) in MeOH (60 mL) was added SOCl.sub.2 (3 mL). The reaction mixture was stirred at rt for 16 h, then refluxed for 3 h. The solvent was removed, to afford Exp-36a (4.58 g, 100%) as a yellow solid: ESI (m/z) [C.sub.9H.sub.12N.sub.2O.sub.2+H].sup.+ 209.

[0683] Preparation of 36b: To a solution of Exp-36a (100 mg, 0.232 mmol), DIPEA (90.0 mg, 0.695 mmol) and HATU (65.1 mg, 0.232 mmol) in DMF (3.0 mL) was added Int-23 (65.1 mg, 0.232 mmol). The reaction mixture was stirred at rt for 2 h. After water (10 mL) was added, the resulting solid was collected by filtration and dried, to afford Exp-36b (133 mg, 92%) as a white solid: ESI (m/z) [C.sub.37H.sub.39N.sub.3O.sub.6+H].sup.+ 622.

[0684] Preparation of 36c: To a solution of Exp-36b (100 mg, 0.168 mmol) in MeOH/IPA/H.sub.2O (1.5 mL/1.5 mL/0.5 mL) was added Pd(OH).sub.2/C (20 mg) and AcOH (0.5 mL). The reaction mixture was stirred under hydrogen (balloon) at 40 C. for 3 h. After filtration, the solvent was removed, to afford Exp-36c (92 mg, 94%) as colorless syrup: ESI (m/z) [C.sub.28H.sub.33N.sub.3O.sub.3+H].sup.+ 460.

[0685] Preparation of Exp-36: To a solution of Exp-36c (480 mg, 0.828 mmol) in DMF (10 mL) was added compound 1 (322 mg, 0.828 mmol) and DIPEA (642 mg, 4.97 mmol). The resulting solution was stirred at 50 C. for 16 h. After the solvent was removed, the residue was twice purified by reverse phase column (10% to 90% of CH.sub.3CN in water followed by 80% of CH.sub.3CN in 0.01% HCl), to afford Exp-36 (309 mg, 50%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 8.33-8.32 (m, 2H), 7.52-7.46 (m, 4H), 7.28 (d, J=8.5 Hz, 2H), 7.21 (d, J=8.5 Hz, 2H), 7.21 (d, J=1.5 Hz, 2H), 4.76-4.72 (m, 1H), 3.69 (s, 3H), 3.36-3.26 (m, 2H), 3.20-3.16 (m, 1H), 2.99-2.95 (m, 1H), 2.87 (t, J=7.0 Hz, 2H), 2.73 (t, J=7.0 Hz, 2H), 2.51 (t, J=7.5 Hz, 2H), 1.81-1.71 (m, 4H); ESI (m/z) [C.sub.34H.sub.38ClN.sub.9O.sub.4+H].sup.+ 672; UPLC AUC=100%; t.sub.R=3.27 min, Method B.

Example 37

Preparation of (S)-2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino) butyl)-[1,1-biphenyl]-4-yl)propanamido)-3-(pyridin-4-yl)propanoic acid (Exp 37)

##STR00406##

[0686] Preparation of Exp-37: To a solution of Exp-36 (152 mg, 0.226 mmol) in THF/MeOH/H.sub.2O (4.0 mL/4.0 mL/4.0 mL) was added NaOH (45.2 mg, 1.13 mmol). The resultant solution was stirred at rt for 3 h. The pH of the solution was adjusted to 2 with 1 N HCl. After the solvent was removed, the residue was purified by reverse phase column (10% to 90% of CH.sub.3CN in water, product came out at 20% to 30%), to afford Exp-37 (76 mg, 46%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 8.68 (d, J=1.5 Hz, 2H), 7.91 (d, J=8.5 Hz, 2H), 7.52-7.48 (m, 4H), 7.28 (d, J=8.0 Hz, 2H), 7.25 (d, J=8.5 Hz, 2H), 4.92-4.83 (m, 1H), 3.56-3.52 (m, 1H), 3.37-3.26 (m, 3H), 2.90-2.86 (m, 2H), 2.73 (t, J=7.0 Hz, 2H), 2.55-2.51 (m, 2H), 1.79-1.73 (m, 4H); ESI (m/z) [C.sub.33H.sub.36ClN.sub.9O.sub.4+H].sup.+ 658; UPLC AUC=100%; t.sub.R=3.11 min, Method B.

Example 38

Preparation of methyl (S)-2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)-3-(piperidin-4-yl)propanoate (Exp 38)

##STR00407##

[0687] Preparation of Exp-38a: To a solution of Exp-36b (800 mg, 1.347 mmol) in MeOH (30 mL) was added Pd(OH).sub.2/C (100 mg) and 3 N HCl in MeOH (2.0 mL) The reaction mixture was stirred under H.sub.2 (55 psi) at rt overnight. LCMS showed reaction completion. After filtration, the solvent was removed to afford Exp-38a (685 mg, 94%) as colorless syrup: ESI (m/z) [C.sub.28H.sub.39N.sub.3O.sub.3+H].sup.+ 466.

[0688] Preparation of Exp-38: To a solution of Exp-38a (685 mg, 1.272 mmol) in DMF (15 mL) was added compound 1 (494 mg, 1.27 mmol) and DIPEA (986 mg, 7.63 mmol). The resulting solution was stirred at 50 C. for 8 h. After the solvent was removed, the residue was twice purified by reverse phase column (10% to 90% of CH.sub.3CN in water, product came out at 20% to 30%), to afford Exp-38 (277 mg, 32%) as yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.51 (m, 4H), 7.30-7.28 (m, 4H), 4.54-4.51 (m, 1H), 3.69 (s, 3H), 3.37-3.34 (m, 4H), 2.99-2.95 (m, 2H), 2.88-2.82 (m, 2H) 2.74-2.71 (m, 2H), 2.62-2.59 (m, 2H), 1.95-1.90 (m, 1H), 1.85-1.54 (m, 8H); 1.39-1.30 (m, 2H); ESI (m/z) [C.sub.34H.sub.44ClN.sub.9O.sub.4+H].sup.+ 678; UPLC AUC=100%; t.sub.R=3.27 min, Method B.

Example 39

Preparation of (S)-2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino) butyl)-[1,1-biphenyl]-4-yl)propanamido)-3-(piperidin-4-yl)propanoic acid (Exp 39)

##STR00408##

[0689] Preparation of Exp-39: To a solution of Exp-38 (73.0 mg, 0.108 mmol) in THF/MeOH/H.sub.2O (2.0 mL/2.0 mL/2.0 mL) was added NaOH (21.5 mg, 0.538 mmol). The resultant solution was stirred at rt for 3 h. The pH of solution was adjusted to 2 with 1 N HCl. After the solvent was removed, the residue was purified by reverse phase column (10% to 90% of CH.sub.3CN in water, product came out at 20% to 30%), to afford Exp-39 (29 mg, 36%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.30-7.27 (m, 4H), 4.36-4.33 (m, 1H), 3.36-3.29 (m, 4H), 2.98-2.94 (m, 2H), 2.84-2.79 (m, 2H) 2.74-2.71 (m, 2H), 2.61-2.58 (m, 2H), 2.06-2.02 (m, 1H), 1.84-1.40 (m, 8H); 1.39-1.25 (m, 2H); ESI (m/z) [C.sub.33H.sub.42ClN.sub.9O.sub.4+H].sup.+ 664; UPLC AUC=96.7%; t.sub.R=3.09 min, Method B.

Example 40

Preparation of methyl (S)-2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)-3-(1-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)piperidin-4-yl)propanoate (Exp 40)

##STR00409##

[0690] Preparation of Exp40a: To a solution of Exp-36c (100 mg, 0.188 mmol) in CH.sub.2Cl.sub.2 (3.0 mL) was added DIPEA (72.8 mg, 0.563 mmol) followed by Boc.sub.2O (45.1 mg, 0.207 mmol) at 0 C. The resulting solution was stirred at 0 C. for 2 h then rt for 2 h. Water (5 mL) was added, and the water phase was extracted with CH.sub.2Cl.sub.2 (5.0 mL). The combined organic phase was dried and concentrated to afford Exp-40a (101 mg, 95%) as a yellow solid: ESI (m/z) [C.sub.33H.sub.41N.sub.3O.sub.5+H].sup.+ 560.

[0691] Preparation of Exp-40b: To a solution of Exp-40a (810 mg, 1.44 mmol) in MeOH (20 mL) was added Pd(OH).sub.2/C (100 mg) and AcOH (1 mL). The resulting suspension was stirred under hydrogen in a par shaker (55 psi) for 72 h. After filtration, the solvent was removed, to afford Exp-40b (875 mg, 97%) as colorless oil: ESI (m/z) [C.sub.33H.sub.47N.sub.3O.sub.5+H].sup.+ 566.

[0692] Preparation of Exp-40c: To a solution of Exp-40b (875 mg, 1.39 mmol) and glucose (504 mg, 2.80 mmol) in MeOH (20 mL) was added NaCNBH.sub.3 (176 mg, 2.80 mmol) and AcOH (168 mg, 2.80 mmol). The resulting solution was stirred at 50 C. for 6 h. Additional glucose (252 mg, 1.39 mmol), NaCNBH.sub.3 (88.0 mg, 1.39 mmol) and AcOH (84.0 mg, 1.39 mmol) were added, and the resulting solution was stirred at 50 C. for 2 h. After the solvent was removed, satd. NaHCO.sub.3 (20 mL) was added. The resulting solid was collected by filtration. The solid was azeotroped with MeOH to remove water affording Exp-40c (935 mg, 92%) as a white solid: ESI (m/z) [C.sub.39H.sub.59N.sub.3O.sub.10+H].sup.+ 730. The material is a borane complex and was used in the next step.

[0693] Preparation of Exp-40d: To a suspension of Exp-40c (800 mg, 1.09 mmol) in MeOH (10 mL) was added 3 N HCl in MeOH (10 mL). The resulting solution was stirred at 40 C. for 2 h. After the solvent was removed, the residue was azeotroped with 0.5 N HCl in MeOH (20 mL) affording Exp-40d (770 mg, 100%) as colorless syrup: ESI (m/z) [C.sub.34H.sub.51N.sub.3O.sub.8+H].sup.+ 630.

[0694] Preparation of Exp-40: To a solution of Exp-40d (770 mg, 1.09 mmol) in DMF (20 mL) was added compound 1 (468 mg, 1.20 mmol) and DIPEA (425 mg, 3.29 mmol). The resulting solution was stirred at 50 C. for 16 h. After the solvent was removed, the residue was purified by reverse phase column (5% to 80% of CH.sub.3CN in water, product came out at 30% to 50%), to afford Exp-40 (556 mg, 60%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.56-7.54 (m, 4H), 7.31-7.29 (m, 4H), 4.52-4.49 (m, 1H), 4.12-4.10 (m, 1H), 3.79-3.44 (m, 10H), 3.40-3.30 (m, 4H), 2.96 (t, J=7.5 Hz, 2H), 2.90-2.80 (m, 1H), 2.75-2.72 (m, 3H), 2.61 (t, J=7.5 Hz, 2H), 2.02-1.96 (m, 1H), 1.81-1.36 (m, 10H); ESI (m/z) [C.sub.40H.sub.56ClN.sub.9O.sub.9+H].sup.+ 842; UPLC AUC=96.2%; t.sub.R=3.15 min, Method B.

Example 41

Preparation of (S)-2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino) butyl)-[1,1-biphenyl]-4-yl)propanamido)-3-(1-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) piperidin-4-yl)propanoic acid (Exp 41)

##STR00410##

[0695] Preparation of Exp-41: To a solution of Exp-40 (214 mg, 0.254 mmol) in MeOH/H.sub.2O (4.0 mL/4.0 mL) was added NaOH (50.8 mg, 1.27 mmol). The resultant solution was stirred at rt for 2 h. The pH was adjusted to 6 with 1 N HCl. After the solvent was removed, the residue was dissolved in CH.sub.3CN/H.sub.2O (2.0 mL/3.0 mL) and the pH was adjusted to 2. The solution was purified by reverse phase column (5% to 80% of CH.sub.3CN in water, product came out at 30% to 50%), to afford Exp-41 (97 mg, 46%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 07.55-7.51 (m, 4H), 7.30-7.28 (D, 4H), 4.35-4.32 (m, 1H), 4.11-4.09 (i, 1H), 3.79-3.63 (i, 6H), 3.40-3.30 (i, 2H), 3.17-2.94 (m, 5H), 2.73-2.54 (m, 6H), 2.02-1.96 (m, 1H), 1.85-1.36 (in, 10H); ESI (m/z) [C.sub.39H.sub.54ClN.sub.9O.sub.9+H]828; UPLC AUC=98.8%; t.sub.R=3.00 min, Method B.

Examples 42-47

[0696] Preparation of Examples 42-47: In schemes analogous to the syntheses of Examples-36-41, (R)-3-amino-2-oxo-4-(pyridin-4-yl)butanoic was used to produce (R)-analogues Examples 42-47:

TABLE-US-00034 Exp-# Structure Spectra 42 [00411] .sup.1H NMR (500 MHz, CD.sub.3OD) 8.50 (d, J = 6.5 Hz, 2H), 8.42 (d, J = 8.5 Hz, 1H), 7.52-7.48 (m, 6H), 7.29-7.21 (m, 4H), 4.90- 4.80 (m, 1H), 3.70 (s, 3H), 3.36- 3.32 (m, 3H), 3.36-3.08 (m, 1H), 2.80 (t, J = 7.5 Hz, 2H), 2.73 (d, J = 7.5 Hz, 2H), 2.53-2.50 (m, 2H), 1.81-1.71 (m, 4H); ESI (m/z) [C.sub.34H.sub.38ClN.sub.9O.sub.4 + H].sup.+ 672; UPLC AUC = 97.1%; t.sub.R = 3.26 min, Method B 43 [00412] .sup.1H NMR (500 MHz, CD.sub.3OD) 8.67 (d, J = 6.5 Hz, 2H), 7.91 (d, J = 6.5 Hz, 2H), 7.52-7.48 (m, 4H), 7.29-7.24 (m, 4H), 4.92- 4.83 (m, 1H), 3.56-3.52 (m, 1H), 3.31-3.23 (m, 3H), 2.90-2.87 (m, 2H), 2.73 (t, J = 7.5 Hz, 2H), 2.55-2.51 (m, 2H), 1.80-1.73 (m, 4H); ESI (m/z) [C.sub.33H.sub.36ClN.sub.9O.sub.4 + H].sup.+ 658; UPLC AUC = 97.1%; t.sub.R = 3.12 min, Method B 44 [00413] .sup.1H NMR (500 MHz, CD.sub.3OD) 8.34 (d, J = 8.0 Hz, 1H), 7.53- 7.51 (m, 4H), 7.30-7.28 (m, 4H), 4.54-4.53 (m, 1H), 3.69 (s, 3H), 3.50-3.40 (m, 4H), 2.99-2.94 (m, 2H), 2.87-2.84 (m, 2H), 2.73 (d, J = 7.0 Hz, 2H), 2.60 (d, J = 7.5 Hz, 2H), 2.02-1.30 (m, 11H); ESI (m/z) [C.sub.34H.sub.44ClN.sub.9O.sub.4 + H].sup.+ 678; UPLC AUC = 95.6%; t.sub.R = 3.27 min, Method B 45 [00414] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.30-7.26 (m, 4H), 4.33-4.31 (m, 1H), 3.50- 3.40 (m, 4H), 2.98-2.94 (m, 2H), 2.82-2.77 (m, 2H), 2.73-2.71 (m, 2H), 2.59 (d, J = 7.5 Hz, 2H), 2.06-2.02 (m, 1H), 1.85-1.68 (m, 6H), 1.55-1.25 (m, 4H); ESI (m/z) [C.sub.33H.sub.42ClN.sub.9O.sub.4 + H].sup.+ 664; UPLC AUC = 94.8%; t.sub.R = 3.12 min, Method B 46 [00415] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.56-7.53 (m, 4H), 7.30-7.29 (m, 4H), 4.51-4.48 (m, 1H), 4.11- 4.07 (m, 1H), 3.79-3.50 (m, 10H), 3.37-3.34 (m, 3H), 3.17- 3.16 (m, 1H), 2.98-2.95 (m, 3H), 2.75-2.70 (m, 4H), 2.62-2.58 (m, 2H), 2.02-1.68 (m, 8H), 1.50- 1.40 (m, 2H); ESI (m/z) [C.sub.40H.sub.56ClN.sub.9O.sub.9 + H].sup.+ 842; UPLC AUC = 94.6%; t.sub.R = 3.14 min, Method B 47 [00416] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.55-7.51 (m, 4H), 7.30-7.28 (m, 4H), 4.34-4.31 (m, 1H), 4.09- 4.07 (m, 1H), 3.79-3.40 (m, 7H), 3.36-3.34 (m, 2H), 3.17-3.12 (m, 2H), 2.98-2.95 (m, 3H), 2.74- 2.72 (m, 4H), 2.62-2.59 (m, 2H), 2.02-1.71 (m, 7H), 1.60-1.40 (m, 3H); ESI (m/z) [C.sub.39H.sub.54ClN.sub.9O.sub.9 + H].sup.+ 828; UPLC AUC = 94.3%; t.sub.R = 3.00 min, Method B

Example 48

Preparation of methyl (S)-2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)-4-(dimethylamino)butanoate (Exp-48)

##STR00417##

[0697] Preparation of Exp-48a: To a solution off Int-23 (50 mg, 0.116 mmol, Int-25 (27.3 mg, 0.139 mmol) and DIPEA (44.9 mg, 0.348 mmol) in DMF (1.0 mL) was added HATU (52.9 mg, 0.139 mmol). The reaction mixture was stirred at room temperature for 2 h. After the solvent was removed, water was added. The white precipitate was filtered and dried, to afford Exp-48a (60 mg, 90%) as a white solid: ESI (m/z) [C.sub.34H.sub.43N.sub.3O.sub.5+H].sup.+ 574.

[0698] Preparation of Exp-48b: To a solution of compound Exp-48a (59 mg, 0.103 mmol) in MeOH (3.0 mL) was added 20% Pd(OH).sub.2 (20 mg). The resultant mixture was stirred under hydrogen (balloon) at room temperature for 8 h. After filtration, the solvent was removed, to afford compound Exp-48b (42 mg, 93%) as colorless oil: ESI (m/z) [C.sub.26H.sub.37N.sub.3O.sub.3+H].sup.+ 440.

[0699] Preparation of Exp-48: To a solution of compound Exp-48b (41.0 mg, 0.093 mmol) in EtOH (0.5 mL) and DMF (0.5 mL) was added compound 1 (39.9 mg, 0.103 mmol) at room temperature. The resultant mixture was stirred under nitrogen at 70 C. for 8 h. The solvent was removed and the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in H.sub.2O, product came out at 30%), to afford compound Exp-48 (52 mg, 85%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.51 (m, 4H), 7.30-7.27 (m, 4H), 4.53-4.51 (m, 1H), 3.72 (s, 3H), 3.36 (t, J=7.0 Hz, 2H), 3.16-2.96 (m, 4H), 2.85 (s, 6H), 2.73 (t, J=7.5 Hz, 2H), 2.63 (t, J=7.5 Hz, 2H), 2.24-2.23 (m, 1H), 2.02-1.97 (m, 1H), 1.81-1.74 (m, 4H); ESI (m/z) [C.sub.32H.sub.42ClN.sub.9O.sub.4+H].sup.+ 652. HPLC AUC=96.0%; t.sub.R=9.69 min, Method A.

Example 49

Preparation of (S)-2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino) butyl)-[1,1-biphenyl]-4-yl)propanamido)-4-(dimethylamino)butanoic acid (Exp-49)

##STR00418##

[0700] Preparation of Exp-49: To a solution of compound Exp-48 (157 mg, 0.241 mmol) in THF (3.0 mL), MeOH (3.0 mL) and water (1.0 mL) was added NaOH (96 mg, 2.41 mmol) at room temperature. The resultant mixture was stirred at room temperature for 3 h. After the solvent was removed, water (3.0 mL) was added. The pH was adjusted to 6 with 1 N HCl and solid precipitation was observed. The solid was collected by filtration, washed with water and dried, to afford compound Exp-49 (144 mg, 94%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.30-7.26 (m, 4H), 4.20-4.18 (m, 1H), 3.34 (t, J=7.0 Hz, 2H), 3.07-2.95 (m, 4H), 2.78 (s, 6H), 2.72 (t, J=7.0 Hz, 2H), 2.63 (t, J=7.5 Hz, 2H), 2.15-2.05 (m, 1H), 1.91-1.74 (m, 5H); ESI (m/z) [C.sub.31H.sub.40ClN.sub.9O.sub.4+H].sup.+ 638; HPLC AUC=96.2%; t.sub.R=9.37 min, Method A.

Example 50

Preparation of N.SUP.6.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysine (Exp-50)

##STR00419##

[0701] Preparation of Exp-50: To a stirred solution of compound Int-23 (2 g, 4.64 mmol) in dichloromethane (30.0 mL) was added TEA (2.60 mL, 18.56 mmol) followed by (L)-NBoc-Lys-OCH.sub.3 (1.60 g, 5.568 mmol) at room temperature. The reaction mixture was stirred at room temperature for 5 min. then 50% T.sub.3P in EtOAc (2.20 mL, 6.96 mmol) was added dropwise. The resulting mixture was stirred at room temperature for 4 h. The reaction mixture was diluted with saturated NaHCO.sub.3 solution (60 mL) and extracted with dichloromethane (250 mL). The combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and the volatiles were removed under reduced pressure to obtained crude which was purified by flash chromatography (0-4% MeOH in dichloromethane) to afford compound Exp-50a (2.30 g, 73%) as an off-white solid: ESI (m/z) [C.sub.39H.sub.51N.sub.3O.sub.7+H].sup.+ 674.

[0702] Preparation of Exp-50b: To a stirred solution of Exp-50a (2.30 g, 3.41 mmol) in IPA (35 mL), water (15 mL) and CH.sub.3CO.sub.2H (0.50 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (600 mg, 50% weight substrate) at room temperature under nitrogen atmosphere. The nitrogen atmosphere was replaced with H.sub.2 (balloon) and the reaction mixture after that which was stirred at room temperature for 16 h. The reaction mixture was filtered through celite pad, the solids were washed with IPA then water. The filtrate was concentrated under reduced pressure to afford Exp-50b (1.70 g, 92%) as an off-white solid: ESI (m/z) [C.sub.31H.sub.45N.sub.3O.sub.5+H].sup.+ 540.

[0703] Preparation of Exp-50c: To a stirred solution of Exp-50b (1.70 g, 3.14 mmol) in DMF (20 mL) was added compound 1 (0.98 g, 3.77 mmol) and DIPEA (3.30 mL, 18.89 mmol) at room temperature under nitrogen. The reaction mixture was stirred at 55 C. for 16 h. The reaction mixture was allowed to cool to room temperature then the volatiles were removed under reduced pressure. The crude was purified by reverse phase (C-18) column chromatography (40-60% CH.sub.3CN in H.sub.2O with 0.1% HCO.sub.2H buffer) to afford Exp-50c (1.0 g, 42%) as an off-white solid. ESI (m/z) [C.sub.37H.sub.50ClN.sub.9O.sub.6+H].sup.+ 752.

[0704] Preparation of Exp-50d: To a stirred solution of Exp-50c (300 mg, 0.398 mmol) in THF (10 mL), MeOH (10 mL) and H.sub.2O (5.0 mL) was added LiOH.Math.H.sub.2O (33 mg, 0.797 mmol) at ambient temperature. The reaction mixture was stirred for 2.5 h at room temperature. The reaction mixture concentrated under reduced pressure; the obtained crude diluted with water (5 mL) then acidified to pH=4 with 2N HCl. The precipitated solids were filtered, washed with water (10 mL) followed by hexanes (30 mL) and dried under reduced pressure to afford Exp-50d (210 mg, 71%) as an off white solid: ESI (m/z) [C.sub.36H.sub.48ClN.sub.9O.sub.6+H].sup.+ 738.

[0705] Preparation of Exp-50: To a stirred solution of Exp-50d (200 mg, 0.270 mmol) in 1,4-dioxane (4.0 mL), was added 4M HCl in 1,4-dioxane (4.0 mL) followed by water (4.0 mL) at room temperature. The resulting solution was stirred for 2 h at room temperature. The reaction mixture was concentrated under reduced pressure. The obtained crude was washed with pentane (220 mL) followed by MTBE (220 mL) and dried under reduced pressure to afford Exp-50 (145 mg, 75%) as light yellow solid: .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.42-7.39 (m, 4H), 7.21-7.15 (m, 4H), 3.84 (t, J=6.2 Hz, 1H), 3.26 (t, J=6.2 Hz, 2H), 3.09 (t, J=6.0 Hz, 2H), 2.85 (t, J=7.8 Hz, 2H), 2.63 (t, J=6.8 Hz, 2H), 2.43 (t, J=7.6 Hz, 2H), 1.91-1.59 (m, 6H), 1.49-1.36 (m, 4H); ESI (m/z) [C.sub.31H.sub.40ClN.sub.9O.sub.4+H].sup.+ 638; HPLC AUC=99.9%; t.sub.R=7.63 min, Method X.

Example 51

Preparation of N.SUP.6.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-D-lysine (Exp-51)

##STR00420##

[0706] Preparation of Exp-51: Example 51 was prepared by substituting (D)-NBoc-Lys-OCH.sub.3 into an analogous sequence of that used to prepare Exp-50: .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.43-7.39 (m, 4H), 7.19-7.16 (m, 4H), 3.84 (t, J=6.2 Hz, 1H), 3.26 (t, J=6.4 Hz, 2H), 3.10-3.01 (m, 2H), 2.84 (t, J=7.8 Hz, 2H), 2.63 (t, J=6.8 Hz, 2H), 2.42 (t, J=7.6 Hz, 2H), 1.91-1.59 (m, 6H), 1.49-1.26 (m, 4H); ESI (m/z) [C.sub.31H.sub.40ClN.sub.9O.sub.4+H].sup.+ 638; HPLC AUC=99.1%; t.sub.R=6.39 min, Method X.

Example 52

Preparation of methyl N.SUP.6.(N.SUP.2.,N.SUP.6.-bis(tert-butoxycarbonyl)-L-lysyl)-N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysinate (Exp-52)

##STR00421##

[0707] Preparation of Exp-52a: To a stirred solution of Int-37 (1.1g, 1.22 mmol) in IPA/water (25 mL, 4:1 ratio) was added 20% Pd(OH).sub.2 on carbon 50% wet (300 mg, 50% weight substrate) at room temperature under nitrogen atmosphere. The nitrogen atmosphere was replaced with H.sub.2 (balloon) and the reaction mixture was stirred for 6 h. The reaction mixture was filtered through a celite pad, washed with IPA, water (20.0 mL, 1:1 ratio) and the filtrate was concentrated under reduced pressure to afford crude compound. The crude material was purified by C-18 reverse phase column chromatography using 40% CH.sub.3CN in H.sub.2O to afford Exp-52a (750 mg, 88%) as an off white solid. ESI (m/z) [C.sub.42H.sub.65N.sub.5O.sub.8+H].sup.+ 768.

[0708] Preparation of Exp-52: To a stirred solution of compound Exp-52a (650 mg, 0.847 mmol) in DMF (5.0 mL) were added compound 1 (270 mg, 0.847 mmol) and DIPEA (218 mg, 1.69 mmol) at ambient temperature under a nitrogen atmosphere. The reaction mixture was warmed to 65 C. and stirred for 15 h. The reaction mixture was cooled, and the volatiles were removed at reduced pressure. The obtained crude product was purified by C-18 reverse phase column chromatography using 40% CH.sub.3CN in H.sub.2O to afford compound Exp-52 (250 mg, 30%) as an off-white powder. .sup.1H NMR (400 MHz, CD.sub.3OD-d.sub.6) 7.56-7.50 (dd, 4H), 7.33-7.25 (dd, 4H), 4.40-4.31 (m, 1H), 3.99-3.89 (m, 1H), 3.70-6.77 (s, 3H), 3.40-3.36 (m, 1H), 3.30-3.26 (m, 1H), 3.21-3.08 (m, 2H), 3.05-2.94 (m, 4H), 2.77-2.696 (t, J=8.0 Hz, 2H), 2.63-2.55 (t, J=8.0 Hz, 2H), 1.82-1.56 (m, 8H), 1.50-1.43 (m, 22H), 1.34-1.24 (m, 4H) ESI (m/z) [C.sub.48H.sub.70ClN.sub.11O.sub.9+H].sup.+ 980; HPLC AUC=86.1%; t.sub.R=7.56 min, Method G.

Example 53

Preparation of methyl N.SUP.6.-(L-lysyl)-N2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysinate (Exp-53)

##STR00422##

[0709] Preparation of Exp-53: To a stirred solution of compound Exp-52 (150 mg, 1.53 mmol) in CH.sub.2Cl.sub.2 (5.0 mL) was added TFA (1 mL) at 0 C. The resulting mixture was stirred at ambient temperature for 4 h. The volatiles were removed under reduced pressure, neutralized with sat. aq. NaHCO.sub.3 solution, and again concentrated under reduced pressure. The residue was purified by C-18 reverse phase column chromatography using 50% CH.sub.3CN in H.sub.2O to afford compound Exp-53 (70 mg, 59%) as an off-white powder. .sup.1H NMR (400 MHz, MeOD-d.sub.6) 7.43-7.38 (m, 4H), 7.21-7.16 (m, 4H), 4.30-4.24 (m, 1H), 3.76-3.66 (m, 1H), 3.59 (s, 3H), 3.28-3.23 (m, 2H), 3.15-3.05 (m, 2H), 2.88-2.81 (m, 4H), 2.65-2.61 (m, 2H), 2.50-2.45 (m, 2H), 1.81-1.54 (m, 10H), 1.46-1.33 (m, 4H), 1.28-1.17 (m, 2H), ESI (m/z) [C.sub.38H.sub.54ClN.sub.11O.sub.5+H].sup.+ 780; HPLC AUC=98.9%; t.sub.R=6.80 min, Method G.

Example 54

Preparation of N.SUP.6.-(L-lysyl)-N2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysine (Exp-54)

##STR00423##

[0710] Preparation of Exp-54: To a stirred solution of compound Exp-53 (200 mg, 0.256 mmol) in THF/H.sub.2O (2:1, 15.0 mL) was added LiOH.Math.H.sub.2O (32 mg, 0.76 mmol) at room temperature. The resulting mixture was stirred at ambient temperature under a nitrogen atmosphere for 4 h. The volatiles were removed under reduced pressure, diluted with water (1.0 mL) and the pH was adjusted to 4 with 2N aqueous HCl. The resulting residue was purified by reverse phase (C-18) column chromatography (30-40% CH.sub.3CN in water; 0.01% HCl as a buffer) to afford compound Exp-54 (167 mg, 80%) as a light yellow powder. .sup.1H NMR (400 MHz, MeOD-d.sub.6) 7.56-7.51 (m, 4H), 7.33-7.29 (d, 4H), 4.29-4.22 (m, 1H), 3.82-3.75 (m, 1H), 3.30-3.23 (m, 2H), 3.17-3.04 (m, 2H), 2.90-2.82 (m, 4H), 2.66-2.58 (m, 2H), 2.53-2.47 (m, 2H), 1.89-1.53 (m, 10H), 1.49-1.35 (m, 4H), 1.32-1.23 (m, 2H); ESI (m/z) [C.sub.37H.sub.52ClN.sub.11O.sub.5+H].sup.+ 766; HPLC AUC=98.6%; t.sub.R=6.25 min, Method J.

Example 55

Preparation of methyl N.SUP.6.((S)-4-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) amino)-2-((tert-butoxycarbonyl)amino)butanoyl)-N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysinate (Exp-55)

##STR00424##

[0711] Preparation of Exp-55a: To a stirred solution of Int-38 (1.30 g, 1.30 mmol) in DMF (5.0 mL) was added piperidine (0.26 mL, 2.61 mmol) at 0 C. The resulting mixture was warmed to ambient temperature and stirred for 6 h. The volatiles were removed under reduced pressure and the obtained crude product was purified by C-18 reverse phase column chromatography using 85% CH.sub.3CN in H.sub.2O to afford Exp-55a (400 mg, 40%) as an off-white solid. ESI (m/z) [C.sub.43H.sub.59N.sub.5O.sub.8+H].sup.+ 774.

[0712] Preparation of Exp-55b: To a stirred solution of Exp-55a (400 mg, 0.52 mmol) in MeOH (6.0 mL) was added D-(+)-glucose (280 mg, 1.55 mmol) followed by acetic acid (0.10 mL, 1.56 mmol) then NaBH.sub.3CN (96 mg, 1.55 mmol) at 0 C. The resulting mixture was stirred at ambient temperature for 15 h. The volatiles were removed under reduced pressure and the obtained crude product was purified by C-18 reverse phase column chromatography using 55% CH.sub.3CN in H.sub.2O, 0.2% acetic acid as a buffer to afford Exp-55b (600 mg, 70%) as an off-white sticky solid. ESI (m/z) [C.sub.55H.sub.83N.sub.5O.sub.18+H].sup.+ 1102.

[0713] Preparation of Exp-55c: To a stirred solution of Exp-55b (600 mg, 0.54 mmol) in IPA/water (11.0 mL, 10:1 ratio) was added 20% Pd(OH).sub.2 on carbon 50% wet (300 mg, 50% weight substrate) at room temperature under nitrogen atmosphere. The nitrogen atmosphere was replaced with H.sub.2 (balloon) and the reaction mixture was stirred for 15 h. The reaction mixture was filtered through celite pad, and the solids were washed with IPA and water (20.0 mL, 1:1 ratio). The combined filtrates were concentrated under reduced pressure to afford Exp-55c (525 mg, 98%) as an off-white sticky solid. ESI (m/z) [C.sub.47H.sub.77N.sub.5O.sub.16+H].sup.+ 968.

[0714] Preparation of Exp-55: To a stirred solution of Exp-55c (525 mg, 0.54 mmol) in DMF (3.0 mL) was added compound 1 (155 mg, 0.59 mmol) and DIPEA (0.60 mL, 3.24 mmol) at ambient temperature under nitrogen atmosphere. The reaction mixture was stirred at 50-60 C. for 15 h. The reaction mixture was cooled, and the volatiles were removed by reduced pressure. The obtained crude product was purified twice by C-18 reverse phase column chromatography using 50% CH.sub.3CN in H.sub.2O, 0.2% acetic acid was a buffer to afford Exp-55 (50 mg, 10%) as a white powder. .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.54 (t, J=8.0 Hz, 4H), 7.31 (d, J=7.6 Hz 4H), 4.40-4.37 (m, 1H), 4.17-4.15 (m, 1H), 3.93-3.90 (m, 2H), 3.83-3.80 (m, 1H), 3.79-3.78 (m, 3H), 3.75-3.73 (m, 2H), 3.70 (s, 3H), 3.68-3.63 (m, 4H), 3.39 (m, 2H), 3.20-3.10 (m, 3H), 2.98 (t, J=7.2 Hz, 2H), 2.76-2.71 (m, 5H), 2.64-2.60 (m, 4H), 1.83-1.62 (m, 7H), 1.46 (s, 11H), 1.36-1.28 (3H); ESI (m/z) [C.sub.53H.sub.82ClN.sub.11O.sub.17+H].sup.+ 1180; HPLC AUC=99.5%; t.sub.R=6.41 min, Method J.

Example 56

Preparation of methyl N.SUP.6.((S)-2-amino-4-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)butanoyl)-N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysinate (Exp-56)

##STR00425##

[0715] Preparation of Exp-56: To a stirred solution of Exp-55 (170 mg, 0.14 mmol) in CH.sub.2Cl.sub.2 (5.0 mL) was added TFA (155 mg, 0.59 mmol) at 0 C. The resulting mixture was stirred at ambient temperature under a nitrogen atmosphere for 6 h. The volatiles were removed under reduced pressure and the obtained TFA salt was diluted with 0.2% aqueous HCl solution (1.0 mL). The volatiles were removed under reduced pressure and this treatment was repeated five times yielding crude HCl salt which was purified by C-18 reverse phase column chromatography using 52% CH.sub.3CN in H.sub.2O, 0.1% HCl was a buffer to afford Exp-56 (65 mg, 42%) as a white powder: .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.43-7.39 (m, 4H), 7.18 (d, J=8.0 Hz, 4H), 4.33-4.29 (m, 1H), 4.13-4.10 (m, 2H), 3.91 (t, J=6.0 Hz, 1H), 3.78-3.70 (m, 2H), 3.69-3.60 (m, 5H), 3.59 (s, 3H), 3.56-3.51 (m, 2H), 3.41-3.29 (m, 6H), 3.27-3.15 (m, 3H), 2.88-2.83 (m, 2H), 2.63 (t, J=7.6 Hz, 2H), 2.49 (t, J=8.6 Hz, 2H), 2.31-2.24 (m, 2H), 1.77-1.45 (m, 10H), 1.35-1.25 (m, 2H); ESI (m/z) [C.sub.48H.sub.74ClN.sub.11O.sub.15+H].sup.+ 1080; HPLC AUC=99.8%; t.sub.R=6.23 min, Method J.

Example 57

Preparation of methyl N.SUP.6.((S)-2-((tert-butoxycarbonyl)amino)-5-methoxy-5-oxopentanoyl)-N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysinate (Exp-57)

##STR00426##

[0716] Preparation of Exp-57a: To a solution of Int-39 (350 mg, 0.428 mmol) in IPA (10 mL) and water (2.0 mL) was added 20% Pd(OH).sub.2 (50 mg). The resultant mixture was stirred under hydrogen (balloon) at 45 C. for 4 h. After filtration, the solvent was removed, to afford Exp-57a (280 mg, 96%) as a white solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 7.51-7.46 (m, 4H), 7.35-7.21 (m, 9H), 6.24 (br s, 2H), 6.13 (d, J=8.0 Hz, 1H), 5.26 (br s, 1H), 5.09 (s, 2H), 4.79 (br s, 1H), 4.59-4.54 (m, 1H), 4.06 (br s, 1H), 3.71 (s, 3H), 3.66 (s, 3H), 3.24-3.16 (m, 4H), 3.02-2.99 (m, 2H), 2.67-2.34 (m, 6H), 2.10-2.09 (m, 1H), 2.06-1.50 (m, 10H), 1.48 (s, 9H), 1.30-1.15 (m, 2H). ESI (m/z) [C.sub.37H.sub.54N.sub.4O.sub.8+H].sup.+ 683.

[0717] Preparation of Exp-57: To a solution of Exp-57a (279 mg, 0.409 mmol) in DMF (3.0 mL) and EtOH (3.0 mL) was added compound 1 (159 mg, 0.409 mmol) at room temperature. The resultant mixture was stirred at 65 C. for 6 h. The solvent was removed, and water (30 mL) was added. The resulting yellow solid was collected by filtration and dried, to afford Exp-57 (328 mg, 90%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.28-7.26 (m, 4H), 4.35-4.32 (m, 1H), 3.99 (br s, 1H), 3.67 (s, 3H), 3.64 (s, 3H), 3.40-3.30 (m, 2H), 3.16-3.05 (m, 2H), 2.96-2.93 (m, 2H), 2.73-2.70 (m, 2H), 2.58 (t, J=7.5 Hz, 2H), 2.39-2.36 (m, 2H), 2.02-1.63 (m, 10H), 1.48 (s, 9H), 1.35-1.20 (m, 2H). ESI (m/z) [C.sub.43H.sub.59ClN.sub.10O.sub.9+H].sup.+ 895: HPLC, AUC=95.1%; t.sub.R=10.20 min, Method A.

Example 58

Preparation of methyl N.SUP.6.((S)-2-amino-5-methoxy-5-oxopentanoyl)-N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysinate (Exp-58)

##STR00427##

[0718] Preparation of Exp-58: To a suspension of Exp-57 (150 mg, 0.168 mmol) in DCM (1.2 mL) was added TFA (0.4 mL). The resultant orange solution was stirred at room temperature for 2 h. The solvent was removed, and the residue was azeotroped with MeOH (2.0 mL) then 1 N HCl in MeOH (32.0 mL) to remove TFA. From the 145 mg of crude obtained, 110 mg of material was purified by reverse phase column (5% to 90% CH.sub.3CN in water, product came out at 40%). The purification failed and 80 mg was recovered. The impure material (80 mg) was dissolved in MeOH (3.0 mL) and thionyl chloride (0.2 mL) was added. The yellow solution was stirred at rt for 2 h. HPLC analysis showed all the carboxylic acid was converted to methyl ester. The solvent removed. The resulting residue was azeotroped with MeOH (33.0 mL) to remove dimethyl sulfide (bp 126 C.). The residue was dissolved in CH.sub.3CN/water and lyophilized, to afford Exp-58 (81 mg) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.28-7.27 (m, 4H), 4.40-4.37 (m, 1H), 3.84 (t, J=6.5 Hz, 1H), 3.69 (s, 3H), 3.68 (s, 3H), 3.35-3.32 (m, 4H), 2.96-2.92 (m, 2H), 2.74-2.71 (m, 2H), 2.57 (t, J=7.5 Hz, 2H), 2.48-2.45 (m, 2H), 2.14-2.02 (m, 2H), 1.84-1.51 (m, 8H), 1.40-1.32 (m, 2H). ESI (m/z) [C.sub.38H.sub.51ClN.sub.10O.sub.7+H].sup.+ 795; HPLC, AUC=97.10%; t.sub.R=10.86 min, Method A.

Example 59

Preparation of N.SUP.6.-((tert-butoxycarbonyl)-L-glutamyl)-N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysine (Exp-59)

##STR00428##

[0719] Preparation of Exp-59: To a solution of Exp-57 (116 mg, 0.130 mmol) in THF (1.5 mL), MeOH (1.5 mL) and water (0.5 mL) was added NaOH (51.8 mg, 1.30 mmol). The resultant solution was stirred at room temperature for 2 h. The reaction mixture was to a pH of 3 with 1 N HCl. The solvent was removed, and water (2.0 mL) was added. The resulting white solid was filtered and further purified by reverse phase column (20% to 90% of CH.sub.3CN in water, product came out at 40% to 60%), to afford compound Exp-59 (92 mg, 82%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.27-7.25 (m, 4H), 4.27-4.25 (m, 1H), 3.99 (br s, 1H), 3.34-3.33 (m, 2H), 3.08-2.94 (m, 4H), 2.71 (t, J=7.5 Hz, 2H), 2.58 (t, J=7.5 Hz, 2H), 2.31 (br s, 2H), 2.01-1.50 (m, 10H), 1.48 (m, 9H), 1.28-1.16 (m, 2H). ESI (m/z) [C.sub.41H.sub.55ClN.sub.10O.sub.9+H].sup.+ 867. HPLC purity 98.0%; HPLC, AUC=98.1%; t.sub.R=9.53 min, Method A.

Example 60

Preparation of N.SUP.6.-(L-glutamyl)-N2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysine (Exp-60)

##STR00429##

[0720] Preparation of Exp-60: To a solution of Exp-57 (178 mg, 0.199 mmol) in MeOH (4.0 mL) was added 4 N HCl (8.0 mL) at room temperature. The resultant mixture was stirred at 40 C. for 2 h. The solvent was removed, the residue was redissolved in 4 N HCl (8.0 mL) and heated at 40 C. for 4 h. The solvent was removed and the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in water, product came out at 30%), to afford Exp-60 (71 mg, 42%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.29-7.26 (m, 4H), 4.39-4.36 (m, 1H), 3.85 (t, J=6.5 Hz, 1H), 3.36-3.20 (m, 4H), 2.95-2.94 (m, 2H), 2.74-2.71 (m, 2H), 2.58 (t, J=7.5 Hz, 2H), 2.47-2.43 (m, 2H), 2.11-2.08 (m, 2H), 1.79-1.60 (m, 6H), 1.54-1.53 (m, 2H), 1.38-1.32 (m, 2H). ESI (m/z) [C.sub.36H.sub.47ClN.sub.10O.sub.7+H].sup.+ 767; HPLC, AUC=95.2%; t.sub.R=9.15 min, Method A.

Example 61

Preparation of methyl N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.((S)-5-methoxy-2-(5-methoxy-5-oxopentanamido)-5-oxopentanoyl)-L-lysinate (Exp-61)

##STR00430##

[0721] Preparation of Exp-61a: To a solution of amine Int-40 (257 mg, 0.341 mmol), 5-methoxy-5-oxopentanoic acid (52.3 mg, 0.358 mmol) and DIPEA (132 mg, 1.02 mmol) in DMF (8.0 mL) was added HATU (126 mg, 0.358 mmol). The reaction mixture was stirred at room temperature for 2 h. After the solvent was removed, water (40 mL) was added. The white precipitate was filtered, dried and purified by silica column (5% to 30% of MeOH in DCM, product came out at 10%), to afford compound Exp-61a (168 mg, 58%) as a white solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 7.51-7.46 (m, 4H), 7.35-7.21 (m, 8H), 6.55 (br s, 1H), 6.40 (br s, 1H), 6.25 (br s, 1H), 5.09 (s, 2H), 4.80 (br s, 2H), 4.58-4.57 (m, 1H), 4.35-4.34 (m, 1H), 3.71 (s, 3H), 3.67 (s, 3H), 3.65 (s, 3H), 3.24-2.88 (m, 6H), 2.67-2.23 (m, 8H), 2.03-1.50 (m, 14H). ESI (m/z) [C.sub.46H.sub.60N.sub.4O.sub.11+H].sup.+ 845.

[0722] Preparation of Exp-61b: To a solution of compound Exp-61a (168 mg, 0.199 mmol) in IPA (5.0 mL) and water (1.0 mL) was added 20% Pd(OH).sub.2 (50 mg) and AcOH (35.8 mg, 0.596 mmol). The resultant mixture was stirred under hydrogen (balloon) at 45 C. for 8 h. After filtration, the solvent was removed, to afford compound Exp-61b (138 mg, 90%) as a white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.49 (m, 4H), 7.29-7.26 (m, 4H), 4.36-4.33 (m, 1H), 4.28-4.25 (m, 1H), 3.68 (s, 3H), 3.65 (s, 3H), 3.64 (s, 3H), 3.10 (t, J=7.0 Hz, 2H), 2.97-2.92 (m, 4H), 2.71 (t, J=7.0 Hz, 2H), 2.58 (t, J=6.0 Hz, 2H), 2.29-2.26 (m, 6H), 2.10-2.00 (m, 1H), 1.98 (s, 3H), 1.97-1.14 (m, 13H). ESI (m/z) [C.sub.38H.sub.54N.sub.4O.sub.9+H].sup.+ 711.

[0723] Preparation of Exp-61: To a solution of compound Exp-61b (136 mg, 0.176 mmol) in DMF (2.0 mL) and EtOH (2.0 mL) was added compound 1 (68.6 mg, 0.176 mmol) and DIEPA (136 mg, 1.05 mmol) at room temperature. The resultant mixture was stirred at 65 C. for 6 h. The solvent was removed, and the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in water, product came out at 40% to 60%), to afford compound Exp-61 (86 mg, 52%) as a yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD) 7.53-7.48 (m, 4H), 7.28-7.25 (m, 4H), 4.33-4.25 (m, 2H), 3.64 (s, 3H), 3.63 (s, 6H), 3.50-3.40 (m, 2H), 3.09 (t, J=6.6 Hz, 2H), 2.94 (t, J=7.5 Hz, 2H), 2.72 (t, J=6.6 Hz, 2H), 2.58 (t, J=7.5 Hz, 2H), 2.37-2.25 (m, 6H), 2.03-1.63 (m, 10H), 1.46-1.24 (m, 4H). ESI (m/z) [C.sub.44H.sub.59ClN.sub.10O.sub.10+H].sup.+ 923; HPLC, AUC=94.1%; t.sub.R=9.79 min, Method A.

Example 62

Preparation of N.SUP.6.-((4-carboxybutanoyl)-D-glutamyl)-N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysine (Exp-62)

##STR00431##

[0724] Preparation of Exp-62a: To a solution of amine Int-40 (50.0 mg, 0.0660 mmol) and DIPEA (25.7 mg, 0.199 mmol) in DMF (8.0 mL) was added dihydro-2H-pyran-2,6(3H)-dione (9.09 mg, 0.0800 mmol). The reaction mixture was stirred at room temperature for 2 h. After the solvent was removed, the residue was dissolved in DCM (5.0 mL) and water (3.0 mL) was added. The pH value of water layer was adjusted to 2 with 1 N HCl. The organic layer was separated, and the water layer was extracted with DCM (3.0 mL). The organic layers were combined, dried, concentrated, to afford Exp-62a (51 mg, 92%) as a white solid: ESI (m/z) [C.sub.45H.sub.58N.sub.4O.sub.11+H].sup.+ 831.

[0725] Preparation of Exp-62b: To a solution of Exp-62a (50.0 mg, 0.060 mmol) in IPA (3.0 mL) and water (0.5 mL) was added 20% Pd(OH).sub.2 (8.0 mg) and AcOH (10.8 mg, 0.181 mmol). The resultant mixture was stirred under hydrogen (balloon) at 45 C. for 8 h. After filtration, the solvent was removed, to afford Exp-62b (42.0 mg, 92%) as a white solid: .sup.1H NMR (300 MHz, CD.sub.3OD) 7.52-7.50 (m, 4H), 7.28-7.25 (m, 4H), 4.33-4.25 (m, 2H), 3.65 (s, 3H), 3.64 (s, 3H), 3.23-2.91 (m, 4H), 2.71 (t, J=6.0 Hz, 2H), 2.58 (t, J=6.0 Hz, 2H), 2.39-2.24 (m, 6H), 2.10-2.00 (m, 1H), 1.96 (m, 6H), 1.89-1.14 (m, 15H).

[0726] Preparation of Exp-62c: To a solution of Exp-62b (41.0 mg, 0.0540 mmol) in DMF (1.0 mL) and EtOH (1.0 mL) was added compound 1 (21.0 mg, 0.0540 mmol) and DIEPA (41.7 mg, 0.324 mmol) at room temperature. The resultant mixture was stirred at 65 C. for 6 h. The solvent was removed, and water was added. The solid was collected by filtration and dried, to afford Exp-62c (42 mg, 85%) as a yellow solid: ESI (m/z) [C.sub.43H.sub.57ClN.sub.10O.sub.10+H].sup.+ 910.

[0727] Preparation of Exp-62: To a solution of Exp-62c (146 mg, 0.158 mmol) in THF (6.0 mL), MeOH (6.0 mL) and water (2.0 mL) was added NaOH (63.2 mg, 1.58 mmol) at room temperature. The resultant mixture was stirred at room temperature for 2 h. The reaction mixture was neutralized with 1 N HCl to pH=3. The solvent was removed, and water (10 mL) was added. The resultant white solid was filtered and suspended in CH.sub.3CN/water (4 mL/4 mL). After 1 N HCl (0.5 mL) was added, a clear solution was observed. The solution was twice purified by reverse phase column (20% to 90% of CH.sub.3CN in water, product came out at 40% to 60%), to afford Exp-62 (93 mg, 66%) as a yellow solid: .sup.1H NMR (300 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.28-7.26 (m, 4H), 4.35-4.27 (m, 2H), 3.11-3.07 (m, 2H), 2.95 (t, J=7.5 Hz, 2H), 2.72 (t, J=7.5 Hz, 2H), 2.60-2.57 (m, 2H), 2.37-2.25 (m, 6H), 2.03-1.20 (m, 16H). ESI (m/z) [C.sub.41H.sub.53ClN.sub.10O.sub.10+H].sup.+ 881. HPLC purity 93.1%; HPLC, AUC=93.1%; t.sub.R=9.77 min, Method A.

Example 63

Preparation of methyl (3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-valyl-L-lysinate (Exp-63)

##STR00432##

[0728] Preparation of Exp-63a: To a stirred solution of Int-53 (1.20 g, 2.2 mmol) in THF (10 mL), MeOH (4.0 mL) and water (4.0 mL) was charged with LiOH.Math.H.sub.2O (277 mg, 6.62 mmol) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure. Obtained crude was diluted with water (20 mL) and acidified to pH=2 with 2N HCl then extracted with EtOAc (350 mL). Combined organic layer washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford Exp-63a (1.05 g, 90%) as an off-white solid: ESI (m/z) [C.sub.32H.sub.38N.sub.2O.sub.5+H].sup.+ 531

[0729] Preparation of Exp-63b: To a stirred solution of Exp-63a (1.00 g, 1.88 mmol) and HATU (0.931 g, 2.45 mmol) in DMF (20 mL) was added Lys(Boc)-OCH.sub.3 (0.67 g, 2.26 mmol) followed by DIPEA (0.93 mL, 5.65 mmol). The reaction mixture was stirred at room temperature for 16 h. The reaction mass was diluted with cold water (50 mL) and the resulting precipitated solid was filtered. Solid was dissolved in EtOAc (150 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford Exp-63b (1.30 g, 74%) as an off-white solid: ESI (m/z) [C.sub.44H.sub.60N.sub.4O.sub.8+H].sup.+ 773.

[0730] Preparation of Exp-63c: To a solution Exp-63b (800 mg, 1.03 mmol) in IPA (20 mL) and EtOAc (20 mL) was added AcOH (0.5 mL) followed by 20% Pd(OH).sub.2 on carbon 50% wet (200 mg, 50% weight substrate) under inert atmosphere. The resultant reaction mixture was purged with N.sub.2, then stirred under H.sub.2 (balloon) for 16 h. The reaction mass was filtered through a celite bed and the filtrate was concentrated under reduced pressure. Obtained crude solid was washed with hexanes (25 mL) and dried under reduced pressure to afford Exp-63c (600 mg, 83%) as an off-white solid; ESI (m/z) [C.sub.36H.sub.54N.sub.4O.sub.6+H].sup.+ 639.

[0731] Preparation of Exp-63d: To a stirred solution of Exp-63c (600 mg, 0.859 mmol) and compound 1 (333 mg, 0.945 mmol) in DMF (15 mL) was added DIPEA (0.42 mL, 2.57 mmol). The reaction mixture was heated at 60 C. and stirred for 16 h. The reaction mass poured into a cold water, the precipitated solid was filtered then washed with water (20 mL). The obtained solid was azeotroped with MeOH (215 mL) and dried to afford Exp-63d (550 mg, 75%) as light brown solid; ESI (m/z) [C.sub.42H.sub.59ClN.sub.10O.sub.7+H].sup.+ 851.

[0732] Synthesis of Exp-63: To a stirred solution of Exp-63d (550 mg, 0.646 mmol) in MeOH (5 mL), was charged with 4M HCl in MeOH (5.0 mL) at room temperature. The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure. Obtained crude was purified by C-18 reverse phase combi-flash chromatography eluting with 10%-100% CH.sub.3CN in water (compound isolated at 45% of CH.sub.3CN) to afford Exp-63 (380 mg, 71%) as a light yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.41-7.38 (m, 4H), 7.19-7.16 (m, 4H), 4.34 (dd, J=9.6, 4.8 Hz, 1H), 3.95 (d, J=8.0 Hz, 1H), 3.60 (s, 3H), 3.26 (t, J=6.8 Hz, 2H), 2.86-2.79 (m, 4H), 2.63 (t, J=7.6 Hz, 2H), 2.53-2.49 (m, 2H), 1.90-1.70 (m, 2H), 1.70-1.55 (m, 7H), 1.43-1.32 (m, 2H), 0.83 (d, J=6.8 Hz, 3H), 0.97 (d, J=6.8 Hz, 3H); ESI (m/z) [C.sub.37H.sub.51ClN.sub.10O.sub.5+H].sup.+ 751; HPLC AUC>99% (R.sub.t=8.73 min); HPLC, AUC=99.0%; t.sub.R=8.73 min, Method V.

Example 64

Preparation of methyl (3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-valyl-L-lysinate (Exp-64)

##STR00433##

[0733] Preparation of Exp-64: To a stirred solution of Exp-63 (200 mg, 0.24 mmol) in THF (5.0 mL), MeOH (2.0 mL) and water (2.0 mL) was charged with LiOH.Math.H.sub.2O (50.9 mg, 1.21 mmol) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure. The obtained crude was diluted with water (10 mL) and pH was adjusted to 2 with 2N HCl. The obtained crude was purified by C-18 reverse phase combi-flash chromatography eluting with 10%-100% CH.sub.3CN in water (compound isolated at 45% of CH.sub.3CN) to afford Exp-64: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.42-7.39 (m, 4H), 7.20-7.18 (m, 4H), 4.34 (dd, J=9.6, 4.8 Hz, 1H), 3.97 (d, J=8.0 Hz, 1H), 3.28 (t, J=3.6 Hz, 2H), 2.87-2.80 (m, 4H), 2.64 (t, J=6.8 Hz, 2H), 2.54-2.48 (m, 2H), 1.92-1.80 (m, 2H), 1.78-1.55 (m, 7H), 1.46-1.38 (m, 2H), 0.85 (d, J=6.8 Hz, 3H), 0.80 (d, J=6.8 Hz, 3H); ESI (m/z) [C.sub.36H.sub.49ClN.sub.10O.sub.5+H].sup.+ 737; HPLC AUC>99% (R.sub.t=8.56 min); HPLC, AUC=99.2%; t.sub.R=8.56 min, Method V.

Example 65

Preparation of methyl (S)-4-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) amino)-2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl) propanamido)butanoate (Exp-65)

##STR00434##

[0734] Preparation of Exp-65a: To a solution of Int-23 (50 mg, 0.116 mmol), Int-24 (57.6 mg, 0.116 mmol) and DIPEA (44.9 mg, 0.348 mmol) and DMF (1.0 mL) was added HATU (48.5 mg, 0.127 mmol). The reaction mixture was stirred at room temperature for 2 h. Solvent was removed and the residue was purified by reverse phase column (10% to 90% of CH.sub.3CN in H.sub.2O, product came out from 30% to 80%), to afford Exp-65a (71 mg, 70%) as white solid: ESI (m/z) [C.sub.44H.sub.63N.sub.3O.sub.15+H].sup.+ 874.

[0735] Preparation of Exp-65b: To a solution of Exp-65a (70 mg, 0.080 mmol) in MeOH (2.0 mL) was added 20% Pd(OH).sub.2 (25 mg). The resultant mixture was stirred under hydrogen (balloon) at room temperature for 8 h. After filtration, the solvent was removed, to afford Exp-65b (53.5 mg, 90%) as colorless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.30-7.25 (m, 4H), 4.52-4.50 (m, 1H), 3.84-3.60 (m, 10H), 2.95-2.87 (m, 4H), 2.72-2.48 (m, 10H), 2.10-2.00 (m, 1H), 1.74-1.64 (m, 5H); ESI (m/z) [C.sub.36H.sub.57N.sub.3O.sub.13+H].sup.+ 740.

[0736] Preparation of Exp-65: To a solution of Exp-65b (50 mg, 0.068 mmol) in EtOH (1.0 mL) and DMF (1.0 mL) was added compound 1 (28.9 mg, 0.074 mmol) at room temperature. The resultant mixture was stirred under nitrogen at 70 C. for 8 h. After the solvent was removed, the residue was triturated with IPA (3.0 mL) and then IPA was decanted, to afford 60 mg of a yellow solid which was purified by reverse phase column (10%90% CH.sub.3CN in H.sub.2O, product came out at 40%), to afford Exp-65 (40 mg, 62%) as an off-white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.49 (m, 4H), 7.29-7.27 (m, 4H), 4.53 (br s, 1H), 3.79-3.62 (m, 16H), 3.50-3.40 (m, 7H), 2.97-2.94 (m, 2H), 2.73 (t, J=7.0 Hz, 2H), 2.61 (t, J=7.0 Hz, 2H), 2.02-1.93 (m, 1H), 1.79-1.72 (m, 5H); ESI (m/z) [C.sub.42H.sub.62ClN.sub.9O.sub.14+H].sup.+ 952. HPLC, AUC=96.1%; t.sub.R=9.08 min, Method A.

Example 66

Preparation of (S)-4-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)-2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)butanoic acid (Exp-66)

##STR00435##

[0737] Preparation of Exp-66: To a solution of compound Exp-65 (266 mg, 0.279 mmol) in dioxane (3.0 mL) and water (3.0 mL) was added LiOH (33.4 mg, 1.39 mmol) at room temperature. The resultant mixture was stirred under nitrogen at room temperature for 1 h. The pH was adjusted to 2 with 1 N HCl, and solid precipitate was collected by filtration, washed with water and dried, to afford compound Exp-66 (233 mg, 79%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.51 (m, 4H), 7.30-7.27 (m, 4H), 4.54-4.51 (m, 1H), 4.17-4.13 (m, 2H), 3.82-3.65 (m, 11H), 3.50-3.45 (m, 2H), 3.42-3.25 (m, 5H), 2.99-2.97 (m, 2H), 2.73 (t, J=7.5 Hz, 2H), 2.62 (t, J=7.5 Hz, 2H), 2.40-2.36 (m, 1H), 2.11-2.09 (m, 1H), 1.78-1.75 (m, 4H); ESI (m/z) [C.sub.41H.sub.60ClN.sub.9O.sub.14+H].sup.+ 938; HPLC, AUC=95.6%; t.sub.R=8.92 min, Method A.

Example 67

Preparation of (S)-3,5-diamino-N(N-(4-(4-(3-(2-amino-3-phenylpropanamido) propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-67)

##STR00436##

[0738] Preparation of Exp-67a: To a stirred solution of Int-17 (0.70 g, 1.68 mmol) and N-Boc phenylalinine (0.44 g, 1.68 mmol) in DMF (10 mL) was added HATU (0.96 g, 2.52 mmol) and diisopropylethylamine (0.32 g, 2.52 mmol) at room temperature under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 16 h. Water (25 mL) was added to reaction mixture followed by extraction with EtOAc (220 mL). The combined organic extracts were concentrated under reduced pressure and the resulting residue was purified by combiflash (40 g column) chromatography eluted with 2% MeOH in dichloromethane to afford Exp-67a (0.50 g, 56%) as an off white solid. ESI (m/z) [C.sub.41H.sub.49N.sub.3O.sub.5+H].sup.+ 664.

[0739] Preparation of Exp-67b: To a stirred solution of Exp-67a (0.50 g, 0.75 mmol) in THF:H.sub.2O (10 mL:3 mL) was added 10% Pd on carbon (0.20 g, 40% wt) at room temperature under nitrogen atmosphere. Then nitrogen was replaced with a H.sub.2 balloon (1 atm) and reaction mixture was stirred for 16 h. The reaction mixture was filtered through celite bed, washed with MeOH and the filtrate was concentrated under reduced pressure to afford Exp-67b (0.40 g, crude) as an off white solid; ESI (m/z) [C.sub.33H.sub.43N.sub.3O.sub.3+H].sup.+ 530.

[0740] Preparation of Exp-67c: To a stirred solution of Exp-67b (0.15 g, 0.28 mmol) in DMF (2 mL) was added compound 1 (0.073 g, 0.28 mmol) and diisopropylethylamine (0.076 g, 0.59 mmol) at room temperature under nitrogen atmosphere. The reaction mixture was warmed to 60 C. and stirred for 16 h. The resulting mixture was cooled to room temperature and concentrated under reduced pressure to get crude product which was purified by reverse phase (C-18) column chromatography using 0-20% CH.sub.3CN in H.sub.2O to afford compound Exp-67c (0.055 g, 26%) as a pale yellow solid. ESI (m/z) [C.sub.39H.sub.48ClN.sub.9O.sub.4+H].sup.+ 742.

[0741] Preparation of Exp-67: To a stirred solution of Exp-67c (0.055 g, 0.07 mmol) in 1,4-dioxane (3 mL) was added 4 M HCl in 1,4-dioxane (0.50 mL, 5 wt/v) at 0 C. The reaction mixture was warmed to room temperature and stirred for 2 h. Reaction mixture was concentrated under reduced pressure and obtained crude was co-distilled with 1,4-dioxane (5 mL) and MTBE (5 mL) then dried under reduced pressure to afford Exp-67 (0.046 g) as a pale brown solid. .sup.1H NMR (400 MHz, MeOD) 7.43-7.39 (m, 4H), 7.27-7.17 (m, 7H), 7.12-7.1 (m, 2H), 3.92-3.88 (t, J=8.0 Hz, 1H), 3.35-3.33 (m, 2H), 3.27-3.26 (m, 2H), 3.05-3.02 (m, 2H), 2.99-2.93 (m, 1H), 2.65-2.61 (t, J=8.0 Hz, 2H), 2.47-2.43 (t, J=8.0 Hz, 2H), 1.67-1.61 (m, 6H); ESI (m/z) [C.sub.34H.sub.40ClN.sub.9O+H].sup.+ 642; HPLC, AUC=91.2%; t.sub.R=10.89 min, Method H.

Examples 68-70

[0742] Preparation of Examples 68-70: In a scheme analogous to the one used for preparation of Example 67, single amino acids were coupled to Int-17 and converted to corresponding Examples 68-70:

TABLE-US-00035 Exp-# AA Structure Spectra 68 D-Boc- Phe [00437] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.42 (t, J = 6.4 Hz, 4H), 7.28-7.18 (m, 7H), 7.13 (d, J = 8.0 Hz, 2H), 3.92 (t, J = 7.6 Hz, 1H), 3.29-3.26 (m, 2H), 3.17-3.14 (m, 1H), 3.09- 2.95 (m, 3H), 2.64 (t, J = 7.2 Hz, 2H), 2.46 (t, J = 7.2 Hz, 2H), 1.70-1.61 (m, 6H); ESI (m/z) [C.sub.34H.sub.40ClN.sub.9O.sub.2 + H].sup.+ 642; HPLC AUC 98.3% (R.sub.t = 8.51 min); HPLC, AUC = 98.3%; t.sub.R = 8.51 min, Method T. 69 Boc- Ala(4- pyridyl) [00438] .sup.1H NMR (500 MHz, CD.sub.3OD) 8.77 (d, J = 6.5 Hz, 2H), 7.87 (d, J = 6.0 Hz, 2H), 7.51-7.49 (m, 4H), 7.28 (d, J = 8.5 Hz, 2H), 7.23 (d, J = 8.0 Hz, 2H), 4.25 (t, J = 7.0 Hz, 1H), 3.44-3.15 (m, 7H), 2.73 (t, J = 7.0 Hz, 1H), 2.61-2.58 (m, 2H), 1.82- 1.75 (m, 6H); ESI (m/z) [C.sub.33H.sub.39ClN.sub.10O.sub.2 + H].sup.+ 643; UPLC purity 95.3%; UPLC, AUC = 95.4%; t.sub.R = 3.05 min, Method B. 70 Boc- Ala(4- pyridyl) [00439] .sup.1H NMR (500 MHz, CD.sub.3OD) 8.70 (d, J = 6.5 Hz, 2H), 7.72 (d, J = 5.5 Hz, 2H), 7.51-7.49 (m, 4H), 7.27 (d, J = 8.0 Hz, 2H), 7.23 (d, J = 8.5 Hz, 2H), 4.20 (t, J = 7.0 Hz, 1H), 3.44-3.15 (m, 7H), 2.73 (t, J = 7.0 Hz, 1H), 2.60-2.57 (m, 2H), 1.93- 1.74 (m, 6H); ESI (m/z) [C.sub.33H.sub.39ClN.sub.10O.sub.2 + H].sup.+ 643; UPLC purity 97.9%; UPLC, AUC = 97.9%; t.sub.R = 3.03 min, Method B.

Example 71

Preparation of (S)-3,5-diamino-N(N-(4-(4-(3-(2-amino-4-(dimethylamino) butanamido)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-71)

##STR00440##

[0743] Preparation of Int-71a: To a stirred solution of Int-25a (350 mg, 2.04 mmol) and Int-17 (850 mg, 2.04 mmol) in CH.sub.2Cl.sub.2 (5.0 mL) was added TEA (0.40 mL, 3.06 mmol) followed by 50% 1-propanephosphonic anhydride solution in EtOAc (1.90 mL, 3.06 mmol). The reaction mixture was stirred under N.sub.2 atmosphere at ambient temperature for 15 hr. The reaction mixture was diluted with CH.sub.2Cl.sub.2 (20.0 mL) and water (20.0 mL), the organic layer was separated, washed with brine solution (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and evaporated under reduced pressure. The obtained crude material was purified by C-18 reverse phase column chromatography using 75% CH.sub.3CN in H.sub.2O to afford Exp-71a (600 mg, 65%) as a white solid. ESI (m/z) [C.sub.38H.sub.52N.sub.4O.sub.5+H].sup.+ 645.

[0744] Preparation of Int-71b: To a stirred solution of Exp-71a (550 mg, 0.85 mmol) in THF/water (10.0 mL, 1:1 ratio) was added 20% Pd(OH).sub.2 on carbon 50% wet (140 mg, 25% weight substrate) at room temperature under nitrogen atmosphere. The reaction mixture was purged with hydrogen then stirred at room temperature under hydrogen (balloon) for 15 h. The reaction mixture was filtered through celite pad and washed with THF and water (20.0 mL, 1:1 ratio) and the filtrate was concentrated under reduced pressure to afford Exp-71b (450 mg, 94%) as a colorless sticky oil. ESI (m/z) [C.sub.30H.sub.46N.sub.4O.sub.3+H].sup.+ 511.

[0745] Preparation of Int-71c: To a stirred solution of Exp-71b (400 mg, 0.78 mmol) in EtOH/DMF (8.80 mL, 10:1 ratio) were added compound 1 (203 mg, 0.78 mmol) and DIPEA (0.80 mL, 4.68 mmol) at ambient temperature under nitrogen atmosphere. The reaction mixture was stirred at 50-60 C. for 15 h. The reaction mixture was cooled, and volatiles were removed under reduced pressure. The obtained crude product was purified twice by C-18 reverse phase column chromatography using 50% CH.sub.3CN in H.sub.2O, 0.1% acetic acid was a buffer to afford Exp-71c (160 mg, 25%) as a pale-yellow solid. ESI (m/z) [C.sub.36H.sub.51ClN.sub.10O.sub.4+H].sup.+ 723.

[0746] Preparation of Int-71: To a stirred solution of Exp-71c (160 mg, 0.22 mmol) in CH.sub.2Cl.sub.2 (2.0 mL) was added 4.0 M HCl in 1,4-dioxane (2.0 mL) at 0 C. The reaction mixture was stirred at ambient temperature for 15 h. The volatiles were removed under reduced pressure and the obtained crude product was purified by C-18 reverse phase column chromatography using 30% CH.sub.3CN in H.sub.2O, 0.1% HCl was used as a buffer to afford Exp-71 (65 mg, 47%) as an off white solid. .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.43-7.40 (m, 4H), 7.20-7.18 (m, 4H), 3.98 (t, J=6.8 Hz, 1H), 3.29-3.26 (m, 4H), 3.16-3.15 (m, 2H), 3.83 (s, 6H), 2.63 (t, J=5.6 Hz, 4H), 2.27-2.17 (m, 2H), 1.86-1.78 (m, 2H), 1.66-1.64 (m, 4H; ESI (m/z) [C.sub.31H.sub.43ClN.sub.10O.sub.2+H].sup.+ 623; HPLC, AUC=99.3%; t.sub.R=7.65 min, Method L.

Example 72

Preparation of 3,5-diamino-N(N-(4-(4-(3-(2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxy hexyl)amino)acetamido)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-72)

##STR00441##

[0747] Preparation of 72a: Acid 2 (213 mg, 1.21 mmol), HATU (462 mg, 1.21 mmol) and DIPEA (314 mg, 2.42 mmol) were dissolved in DMF (6.0 mL), and the reaction mixture was stirred at room temperature for 30 min. A suspension of amine Int-17 (550 mg, 1.21 mmol) and DIPEA (314 mg, 2.42 mmol) in DMF (4.0 mL) was added dropwise to the above solution, and the reaction mixture was stirred at room temperature for 2 h. The solvent was removed, water (30 mL) was added, and the precipitated solid was collected by filtration, washed with water (20 mL) and dried to afford Exp-72a (602 mg, 86%) as white solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 7.48-7.47 (m, 4H), 7.35-7.20 (m, 9H), 6.10 (br s, 1H), 5.09 (s, 2H), 5.00 (br s, 1H), 4.60 (br s, 1H), 3.73 (d, J=6.0 Hz, 2H), 3.33 (t, J=7.0 Hz, 2H), 3.23 (t, J=6.0 Hz, 2H), 2.70-2.64 (m, 4H), 1.90-1.87 (m, 2H), 1.69-1.59 (m, 4H), 1.45 (s, 9H); ESI (m/z) [C.sub.34H.sub.43N.sub.3O.sub.5+H].sup.+ 574.

[0748] Preparation of 72b: To a solution of Int-72a (600 mg, 1.04 mmol) in THF (12 mL) was added 6N HCl in IPA (12 mL) at room temperature. Solid precipitation was observed after 2 min. The reaction mixture was stirred at room temperature for 2 h. The solvent was removed to afford Exp-72b (529 mg, 99%) as white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.51-7.50 (m, 4H), 7.33-7.26 (m, 9H), 5.05 (s, 2H), 6.10 (br s, 1H), 3.64 (s, 2H), 3.32-3.29 (m, 2H), 3.14 (t, J=7.5 Hz, 2H), 2.70-2.60 (m, 4H), 1.95-1.90 (m, 2H), 1.70-1.50 (m, 4H); ESI (m/z) [C.sub.29H.sub.35N.sub.3O.sub.3+H].sup.+ 474.

[0749] Preparation of 72c: To a solution of Exp-72b (525 mg, 1.03 mmol) in MeOH (15 mL) was added D-glucose (556 mg, 3.09 mmol) followed by AcOH (185 mg, 3.09 mmol) and NaCNBH.sub.3 (194 mg, 3.09 mmol). The resulting solution was stirred at 65 C. for 3 h. Additional D-glucose (370 mg, 2.06 mmol), AcOH (123 mg, 2.06 mmol) and NaCNBH.sub.3 (129 mg, 2.06 mmol) were added, and the reaction mixture was heated at 65 C. and stirred for 2 h. Solvent was removed, the residue was azeotroped with 2 N HCl in MeOH (210 mL) then purified by reverse phase column (5% to 80% of CH.sub.3CN in H.sub.2O, product came out at 40%), to afford Exp-72c (648 mg, 75%) as white syrup: .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.63 (br s, 1H), 8.49 (br s, 1H), 7.56-7.53 (m, 4H), 7.35-7.24 (m, 9H), 5.00 (s, 2H), 4.09-4.01 (m, 4H), 3.70-3.00 (m, 18H), 2.65-2.49 (m, 4H), 1.78-1.75 (m, 2H), 1.59-1.56 (m, 2H), 1.45-1.42 (m, 2H); ESI (m/z) [C.sub.41H.sub.59N.sub.3O.sub.13+H].sup.+ 802.

[0750] Preparation of 72d: To a solution of Exp-72c (648 mg, 0.773 mmol) in MeOH (16 mL) and water (8.0 mL) was added 20% Pd(OH).sub.2 (100 mg). The resultant mixture was stirred under hydrogen (balloon) at room temperature for 6 h. After filtration, the solvent was removed to afford Exp-72d (516 mg, 95%) as a colorless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.55-7.52 (m, 4H), 7.31-7.29 (m, 4H), 4.30-4.10 (m, 4H), 3.80-3.37 (m, 16H), 3.00-2.96 (m, 2H), 3.00-2.96 (m, 2H), 2.74-2.72 (m, 4H), 1.95-1.90 (m, 2H), 1.85-1.70 (m, 4H); ESI (m/z) [C.sub.33H.sub.53N.sub.3O.sub.11+H].sup.+ 668. Preparation of 72: To a solution of Exp-72d (516 mg, 0.733 mmol) in EtOH (10 mL) and DMF (10 mL) was added compound 1 (313 mg, 0.806 mmol) at room temperature. The resultant mixture was stirred under nitrogen at 70 C. for 8 h. The solvent as removed and the residue was titurated with EtOH forming a solid. The solid was collected by filtration then dissolved in water/CH.sub.3CN and purified by reverse phase column (5%80% of CH3CN/0.5% AcOH in water, product came out at 40%), to afford Exp-72 (514 mg, 70%) as yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.42-7.37 (m, 4H), 7.18-7.16 (m, 4H), 3.80-3.79 (m, 2H), 3.69-3.51 (m, 10H), 3.37-3.08 (m, 6H), 2.64-2.59 (m, 8H), 1.84 (s, 6H), 1.81-1.63 (m, 6H); ESI (m/z) [C.sub.39H.sub.58ClN.sub.9O.sub.12+H].sup.+ 880. HPLC, AUC=99.4%; t.sub.R=9.24 min, Method A.

Example 73

Preparation of 3,5-diamino-N(N-(4-(4-(3-((S)-2-amino-4-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)butanamido)propyl)-[1,1-biphenyl]-4-yl)butyl) carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-73)

##STR00442##

[0751] Preparation of Exp-73a: To a solution of acid Int-26 (29.2 mg, 0.066 mmol) in DMF (1.0 mL) was added amine Int-17 (30.0 mg, 0.066 mmol), HATU (25.2 mg, 0.066 mmol) and DIPEA (25.7 mg, 0.199 mmol). The reaction mixture was stirred at room temperature for 1 h. After the solvent was removed, the residue was partitioned with EtOAc (2.0 mL) and H.sub.2O (2.0 mL). The water layer was extracted with EtOAc (2.0 mL2). The organic layers were combined and concentrated, to afford Exp-73a (49 mg, 88%) as a white solid: ESI (m/z) [C.sub.51H.sub.58N.sub.4O.sub.7+H].sup.+ 839.

[0752] Preparation of Exp-73b: To a solution of Exp-73a (341 mg, 0.406 mmol) in CH.sub.2Cl.sub.2 (3.0 mL) was added piperidine (1.0 mL). The resultant mixture was stirred at room temperature for 2 h. After the solvent was removed, the residue was washed with hexanes (5.0 mL3), to afford Exp-73b (230 mg, 92%) as white solid: ESI (m/z) [C.sub.36H.sub.48N.sub.4O.sub.5+H].sup.+ 617.

[0753] Preparation of Exp-73c: To a solution of Exp-73b (40 mg, 0.065 mmol) in MeOH (1.2 mL) was added D-glucose (35 mg, 0.195 mmol) followed by AcOH (11.6 mg, 0.195 mmol) and NaCNBH.sub.3 (12.2 mg, 0.195 mmol). The resulting solution was stirred at 60 C. for 3 h. Additional D-glucose (23.3 mg, 0.130 mmol), AcOH (7.70 mg, 0.130 mmol) and NaCNBH.sub.3 (8.10 mg, 0.130 mmol) were added, and the reaction mixture was warmed to 60 C. and stirred for 2 h. Solvent was removed, and the residue was dissolved in water (4.0 mL) and AcOH (0.5 mL). The solution was purified by reverse phase column (10% to 90% of CH.sub.3CN in H.sub.2O, product came out at 40%), to afford Exp-73c (52 mg, 80%) as colourless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.51-7.49 (m, 4H), 7.33-7.25 (m, 9H), 5.05 (s, 2H), 4.15 (br s, 1H), 4.05-3.95 (m, 2H), 3.79-3.64 (m, 10H), 3.25-2.80 (m, 10H), 2.70-2.60 (m, 4H), 2.10-2.00 (m, 1H), 1.97 (s, 10H), 1.95-1.85 (m, 3H), 1.70-1.50 (m, 4H); 1.45 (s, 9H); ESI (m/z) [C.sub.48H.sub.72N.sub.4O.sub.15+H].sup.+ 945.

[0754] Preparation of Exp-73d: To a solution of Exp-73c (280 mg, 0.279 mmol) in MeOH (8.0 mL) was added 20% Pd(OH).sub.2 (100 mg). The resultant mixture was stirred under hydrogen (balloon) at room temperature for 16 h. After filtration, the solvent was removed, to afford Exp-73d (214 mg, 88%) as colorless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.27-7.25 (m, 4H), 4.15 (br s, 1H), 3.90-3.87 (m, 2H), 3.79-3.60 (m, 10H), 2.95-2.91 (m, 4H), 2.74-2.61 (m, 10H), 2.00-1.99 (m, 1H), 1.91 (s, 6H), 1.87-1.66 (m, 7H), 1.45 (s, 9H); ESI (m/z) [C.sub.40H.sub.66N.sub.4O.sub.13+H].sup.+ 811.

[0755] Preparation of Exp-73e: To a solution of Exp-73d (170 mg, 0.195 mmol) in EtOH (1.5 mL) and DMF (1.5 mL) was added compound 1 (83.0 mg, 0.215 mmol) at room temperature. The resultant mixture was stirred under nitrogen at 70 C. for 8 h. After solvent removed, the residue was triturated with IPA (3.0 mL) then the IPA was decanted. The residue was dried under high vacuum to afford Exp-73e (172 mg, 86%) as yellow solid: ESI (m/z) [C.sub.46H.sub.71ClN.sub.10O.sub.14+H].sup.+ 1023.

[0756] Preparation of Exp-73: To a solution of Exp-73e (170 mg, 0.166 mmol) in EtOH (2.0 mL) was added 4 N HCl (6.0 mL) at room temperature. The resultant mixture was stirred at room temperature for 8 h. The solvent was removed and the residue was directly purified by reverse phase column (5% to 90% of CH.sub.3CN in H.sub.2O, the product came out at 30% to 80%), to afford Exp-73 (119 mg, 68%) as yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.28-7.25 (m, 4H), 4.05-3.97 (m, 3H), 3.79-3.62 (m, 10H), 3.37-3.26 (m, 4H), 2.98-2.54 (m, 10H), 2.20-2.19 (m, 1H), 1.91-1.75 (m, 7H); ESI (m/z) [C.sub.41H.sub.63ClN.sub.10O.sub.12+H].sup.+ 923; HPLC, AUC=98.3%; t.sub.R=8.81 min, Method A.

Example 74

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(3-(2-((4-(3-(((2R,3S,4S,5S)-2,3,4,5,6-pentahydroxyhexyl)((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)propyl) phenyl)amino)acetamido)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl) pyrazine-2-carboxamide (Exp-74)

##STR00443##

[0757] Preparation of Exp-74a: To a stirred solution of compound Int-18 (200 mg, 0.423 mmol) and compound Int-59 (201 mg, 0.423 mmol) in MeOH (20 mL) was added AcOH (0.026 mL, 0.423 mmol) followed by NaCNBH.sub.3 (26 mg, 0.423 mmol). The resultant mixture was stirred at room temperature for 2 h. The volatiles were removed under reduced pressure and crude product was purified by C-18 reverse phase column chromatography using 20-40% CH.sub.3CN in H.sub.2O to afford Exp-74a (140 mg, 35%) as a white solid. ESI (m/z) [C.sub.50H.sub.70N.sub.4O.sub.13+H].sup.+ 935

[0758] Preparation of Exp-74b: To a stirred solution of Exp-74a (140 mg, 0.149 mmol) in MeOH (14 mL) and water (4 mL) was added 10% Pd/C on carbon (40 mg, 50% weight by weight) at room temperature under nitrogen atmosphere. The reaction mixture was purged with H.sub.2 then stirred under H.sub.2 (balloon) for 6 h. The reaction mixture was filtered through celite pad and washed with MeOH (20 mL). The filtrate was concentrated under reduced pressure and the crude product was purified by C-18 reverse phase column chromatography using 40% CH.sub.3CN in H.sub.2O to afford Exp-74b (70 mg, 58%) as a gummy solid. ESI (m/z) [C.sub.42H.sub.64N.sub.4O.sub.11+H].sup.+ 801.

[0759] Preparation of Exp-74: To a stirred solution of Exp-74b (300 mg, 0.375 mmol) in DMF (5 mL) and EtOH (5 mL) was added compound 1 (98 mg, 0.375 mmol) and DIPEA (0.387 mg, 3.01 mmol) at room temperature under nitrogen. The reaction mixture was stirred at 80 C. for 12 h. The reaction mixture was allowed to cool, and volatiles were removed under reduced pressure. The crude product was purified by preparative HPLC to afford compound Exp-74 (40 mg, 12%) as off white solid. .sup.1H NMR (400 MHz, D.sub.2O-d.sub.3) 8.32 (s, 1H), 7.29-7.24 (m, 2H), 7.13-7.04 (m, 4H), 6.87-6.85 (d, J=8.0 Hz 2H), 6.74-6.72 (m, 2H), 6.45-6.43 (d, J=8.0 Hz 2H), 3.97-3.86 (m, 2H), 3.7-3.68 (m, 1H), 3.67-3.64 (m, 1H), 3.63-3.58 (m, 6H), 3.54-3.48 (m, 2H), 3.46-3.41 (m, 2H), 3.15-2.95 (m, 1H), 2.55-2.45 (m, 2H), 2.44-2.26 (m, 2H), 2.24-2.16 (m, 2H), 1.81-1.69 (m, 2H), 1.65-1.54 (m, 2H), 1.51-1.40 (m, 4H) ESI (m/z) [C.sub.48H.sub.69ClN.sub.10O.sub.12].sup.+ 1013; HPLC, AUC=98.1%; t.sub.R=7.00 min, Method E.

Example 75

Preparation of 3,5-diamino-N(N-(4-(4-(3-(2-((2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)amino)acetamido)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-75)

##STR00444##

[0760] Preparation of Exp-75a: To a stirred solution of Int-18 (800 mg, 1.69 mmol) and compound Int-22 (860 mg, 2.02 mmol) in MeOH (70 mL) was added AcOH (0.2 mL, 3.38 mmol). The reaction mixture was heated to reflux and stirred for 12 h. The reaction mixture was cooled to room temperature, NaCNBH.sub.3 (180 mg, 1.69 mmol) was added and resulting mixture was stirred at room temperature for 6 h. The volatiles were removed under reduced pressure and crude product was purified by C-18 reverse phase column chromatography using 20-40% CH.sub.3CN in H.sub.2O to afford Exp-75a (440 mg, 22%) as a white solid. ESI (m/z) [C.sub.43H.sub.64N.sub.4O.sub.13+H].sup.+ 845.

[0761] Preparation of Exp-75b: To a stirred solution of Exp-75a (400 mg, 0.473 mmol) in THF (30 mL), water (10 mL) and acetic acid (0.10 mL) was added 10% Pd/C on carbon (40 mg, 50% weight by weight) at room temperature under nitrogen atmosphere. The reaction mixture was purged with H.sub.2 then stirred under hydrogen (balloon) for 12 h. The reaction mixture was filtered through celite pad and the solids were washed with MeOH (20 mL). The combined filtrate was concentrated under reduced pressure and the crude product was purified by C-18 reverse phase column chromatography using 40% CH.sub.3CN in H.sub.2O to afford Exp-75b (170 mg, 50%) as a gummy solid. ESI (m/z) [C.sub.35H.sub.58N.sub.4O.sub.11+H].sup.+ 711.

[0762] Preparation of Exp-75: To a stirred solution of Exp-75b (170 mg, 0.239 mmol) in DMF (10 mL) was added compound 1 (140 mg, 0.358 mmol) and DIPEA (246 mg, 1.91 mmol) at room temperature under nitrogen. The reaction mixture was warmed to 80 C. and stirred for 12 h. The reaction mixture was allowed to cool to room temperature, the volatiles were removed under reduced pressure. The resulting residue was treated with IPA (15 mL) and to form a solid precipitate which was collected by filtration. The obtained crude compound was purified by preparative HPLC to afford Exp-75 (62 mg, 28%) as an off white solid. .sup.1H NMR (400 MHz, D.sub.2O)) 8.35 (m, 1H), 7.51-7.49 (d, J=8.0 Hz, 2H), 7.38-7.36 (d, J=8.0 Hz, 2H), 7.27-7.25 (d, J=8.0 Hz, 2H), 7.14-7.12 (d, J=8.0 Hz, 2H), 3.93-3.87 (m, 2H), 3.75-3.71 (m, 1H), 3.70-3.68 (m, 2H), 3.67-3.65 (m, 3H), 3.58-3.45 (m, 7H), 3.26-3.11 (m, 4H), 3.06-2.90 (m, 3H), 2.88-2.78 (m, 4H), 2.61-2.55 (m, 4H), 1.84-1.71 (m, 4H), 1.67-1.61 (m, 2H). ESI (m/z) [C.sub.41H.sub.63ClN.sub.10O.sub.12+H].sup.+ 923: HPLC, AUC=85.1%; t.sub.R=7.10 min, Method F.

Example 76

Preparation of hexyl ((S)-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) amino)-1-((3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propyl)amino)-1-oxohexan-2-yl)carbamate (Exp-76)

##STR00445##

[0763] Preparation of Exp-76a: To a solution of Int-29 (50.0 mg, 0.0530 mmol) in THF (20 mL), MeOH (1.5 mL) and water (0.5 mL) was added NaHCO.sub.3 (13.3 mg, 0.159 mmol) and n-hexylchloroformate (13.0 mg, 0.0790 mmol) at 0 C. The resultant mixture was stirred at 0 C. for 30 min and room temperature for 1 h. After the solvent was removed, water was added, and the water layer was decanted. The residue was dried, to afford Exp-76a (48 mg, 91%) as colorless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.50-7.47 (m, 4H), 7.30-7.21 (m, 9H), 5.05 (m, 2H), 4.06 (br s, 3H), 3.84-3.43 (m, 14H), 3.25-3.13 (m, 2H), 2.66-2.61 (m, 8H), 1.85-1.51 (m, 12H); ESI (m/z) [C.sub.52H.sub.80N.sub.4O.sub.15+H].sup.+ 1001.

[0764] Preparation of Exp-76b: To a solution of Exp-76a (255 mg, 0.255 mmol) in EtOH (10 mL) and water (5.0 mL) was added 20% Pd(OH).sub.2 (30 mg) and AcOH (92 mg, 1.528 mmol). The resultant mixture was stirred under hydrogen (balloon) at room temperature at 45 C. for 4 h. After filtration, the solvent was removed, to afford Exp-76b (210 mg, 84%) as white syrup: ESI (m/z) [C.sub.44H.sub.74N.sub.4O.sub.13+H].sup.+ 867.

[0765] Preparation of Exp-76: To a solution of Exp-76b (168 mg, 0.170 mmol) in DMF (5.0 mL) was added compound 1 (72.7 mg, 0.187 mmol) and DIPEA (132 mg, 1.02 mmol) at room temperature. The resultant mixture was stirred at 60 C. for 16 h. After the solvent was removed, the residue was purified by reverse phase column [10% to 80% of CH.sub.3CN in water (0.5% AcOH buffered), product came out at 40%], to afford Exp-76 (89 mg, 48%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.47 (m, 4H), 7.28-7.24 (m, 4H), 4.03-4.02 (m, 3H), 3.87-3.60 (m, 12H), 3.26-3.21 (m, 4H), 2.74-2.65 (m, 10H), 1.89-1.28 (m, 20H), 0.89-0.87 (m, 3H); ESI (m/z) [C.sub.50H.sub.79ClN.sub.10O.sub.14+H].sup.+ 1079; HPLC, AUC=94.0%; t.sub.R=10.04 min, Method A.

Example 77

Preparation of tert-butyl ((S)-1-(((S)-1-(((S)-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)-1-((3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propyl)amino)-1-oxohexan-2-yl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)carbamate I (Exp-77)

##STR00446##

[0766] Preparation of Exp-77a: To a solution of Int-29 (50.0 mg, 0.0530 mmol) in DMF (2.0 mL) was added HATU (24.1 mg, 0.0630 mmol), DIPEA (27.3 mg, 0.211 mmol) and N-Boc-Ala-Ala-OH (15.1 mg, 0.0580 mmol) at room temperature. The resultant mixture was stirred at room temperature for 1 h. After the solvent was removed, the residue was purified by reverse phase column (5% to 100% of CH.sub.3CN in water, product came out at 80%], to afford Exp-77a (369 mg, 61%) as colorless syrup: .sup.1H NMR (300 MHz, CD.sub.3OD) 7.51-7.47 (m, 4H), 7.33-7.24 (m, 9H), 5.05 (s, 2H), 4.34-4.24 (m, 2H), 4.11-3.97 (m, 3H), 3.82-3.63 (m, 10H), 3.50-3.14 (m, 10H), 2.67-2.64 (m, 4H), 1.88-1.44 (m, 12H), 1.43 (m, 9H), 1.40-0.87 (m, 6H); ESI (m/z) [C.sub.56H.sub.86N.sub.6O.sub.17+H].sup.+ 1115.

[0767] Preparation of Exp-77b: To a solution of Exp-77a (220 mg, 0.197 mmol) in EtOH (2.0 mL) and water (4.0 mL) was added 20% Pd(OH).sub.2 (30 mg) and AcOH (71.1 mg, 1.18 mmol). The resultant mixture was stirred under hydrogen (balloon) at room temperature at 45 C. for 4 h. After filtration, the solvent was removed, to afford Exp-77b (205 mg, 94%) as colorless syrup: ESI (m/z) [C.sub.44H.sub.74N.sub.4O.sub.13+H].sup.+ 867.

[0768] Preparation of Exp-77: To a solution of Exp-77b (164 mg, 0.149 mmol) in DMF (2.0 mL) and EtOH (2.0 mL) was added compound 1 (57.9 mg, 0.149 mmol) and DIPEA (115 mg, 0.893 mmol) at room temperature. The resultant mixture was stirred at 60 C. for 16 h. After the solvent was removed, the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in water, product came out at 50%), to afford Exp-77 (120 mg, 67%) as yellow syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.29-7.25 (m, 4H), 4.34-4.32 (m, 2H), 4.27-4.25 (m, 2H), 4.17-4.16 (m, 1H), 4.03-3.64 (m, 10H), 3.39-3.24 (m, 10H), 2.75-2.66 (m, 4H), 1.88-1.75 (m, 10H), 1.45 (m, 9H), 1.44-1.36 (m, 8H); ESI (m/z) [C.sub.54H.sub.85ClN.sub.12O.sub.16+H].sup.+ 1193; HPLC, AUC=98.8%; t.sub.R=9.58 min, Method A.

Example 78

Preparation of 3,5-diamino-N(N-(4-(4-(3-((S)-2-((S)-2-((S)-2-aminopropanamido) propanamido)-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)hexanamido) propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-78)

##STR00447##

[0769] Preparation of Exp-78: To a solution of Exp-77 (62 mg, 0.0520 mmol) in EtOH (1.0 mL) was added 4 N HCl (2.0 mL) at room temperature. The resultant mixture was stirred at 45 C. for 2 h. The solvent was removed to afford Exp-78 (60 mg, 96%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.29-7.25 (m, 4H), 4.37-4.33 (m, 2H), 4.19-4.16 (m, 2H), 3.94-3.93 (m, 1H), 3.84-3.65 (m, 10H), 3.45-3.23 (m, 10H), 2.75-2.66 (m, 4H), 1.87-1.75 (m, 10H), 1.54-1.40 (m, 8H); ESI (m/z) [C.sub.49H.sub.77ClN.sub.12O.sub.14+H].sup.+ 1093; HPLC, AUC=96.9%; t.sub.R=8.0784 min, Method A.

Example 79

Preparation of tert-butyl ((S)-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) amino)-1-((3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propyl)amino)-1-oxohexan-2-yl)carbamate (Exp-79)

##STR00448##

[0770] Preparation of Exp-79: To a solution of compound Int-28 (370 mg, 0.386 mmol) in DMF (8.0 mL) was added compound 1 (240 mg, 0.617 mmol) and DIPEA (299 mg, 2.31 mmol) at room temperature. The resultant mixture was heated to 65 C. and stirred for 16 h and then heated to 70 C. and stirred for 16 h. After the solvent was removed, the residue was purified by reverse phase column [10% to 80% of CH.sub.3CN in water (0.5% AcOH buffered), product came out at 40%], to afford compound Exp-79 (43 mg, 10%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.47 (m, 4H), 7.28-7.24 (m, 4H), 3.98 (br s, 3H), 3.79-3.61 (m, 10H), 3.36-3.16 (m, 6H), 2.85-2.65 (m, 8H), 1.94 (s, 2.7H), 1.85-1.45 (m, 12H), 1.44 (s, 9H); ESI (m/z) [C.sub.48H.sub.75ClN.sub.10O.sub.14+H].sup.+ 1051; HPLC, AUC=94.4%; t.sub.R=9.68 min, Method A.

Example 80

Preparation of 3,5-diamino-N(N-(4-(4-(3-((S)-2-amino-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)hexanamido)propyl)-[1,1-biphenyl]-4-yl)butyl) carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-80)

##STR00449##

[0771] Preparation of Exp-80: To a solution of Exp-79 (83 mg, 0.079 mmol) in EtOH (2.0 mL) was added 4 N HCl (4.0 mL) at room temperature. The resultant mixture was stirred at room temperature for 16 h. After the solvent was removed, the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in water, product came out from 40%), to afford Exp-80 (60 mg, 74%) as an off-white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.55-7.53 (m, 4H), 7.31-7.29 (m, 4H), 4.20-4.19 (m, 2H), 3.87-3.66 (m, 11H), 3.47-3.36 (m, 10H), 2.77-2.71 (m, 4H), 1.95-1.77 (m, 10H), 1.53-1.52 (m, 2H); ESI (m/z) [C.sub.43H.sub.67ClN.sub.10O.sub.12+H].sup.+ 951; HPLC, AUC=98.0%; t.sub.R=8.73 min, Method A.

Example 81

Preparation of 3,5-diamino-N(N-(4-(4-(3-((S)-2-((S)-2-amino-4-methyl pentanamido)-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)hexanamido) propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-81)

##STR00450##

[0772] Preparation of Exp-81: In three steps Exp-81 was synthesized beginning with Int-29 and N-Boc-Lue-OH using a sequence analogous to the one used to prepare Exp-78: .sup.1H NMR (500 MHz, D.sub.2O) 7.55 (d, J=8.0 Hz, 2H), 7.42 (d, J=8.0 Hz, 2H), 7.32 (d, J=8.0 Hz, 2H), 7.18 (d, J=8.0 Hz, 2H), 4.26-4.23 (m, 1H), 4.15-4.12 (m, 2H), 3.95-3.92 (m, 1H), 3.78-3.58 (m, 10H), 3.32-3.20 (m, 10H), 2.65-2.59 (m, 4H), 1.86-1.59 (m, 13H), 1.38-1.34 (m, 2H), 0.88-0.87 (m, 6H); ESI (m/z) [C.sub.49H.sub.78ClN.sub.11O.sub.13+H].sup.+ 1064; HPLC, AUC=99.08%; t.sub.R=8.92 min, Method A.

Example 82

Preparation of 3,5-diamino-N(N-(4-(4-(3-((S)-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)-2-((S)-2,6-diaminohexanamido)hexanamido)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-82)

##STR00451##

[0773] Preparation of Exp-82: In three steps Exp-82 was synthesized beginning with Int-29 and N-Boc-Lys(Boc)-OH using a sequence analogous to the one used to prepare Exp-78: .sup.1H NMR (500 MHz, D.sub.2O) 7.57 (d, J=8.0 Hz, 2H), 7.45 (d, J=8.0 Hz, 2H), 7.34 (d, J=8.0 Hz, 2H), 7.20 (d, J=8.0 Hz, 2H), 4.23-4.20 (m, 1H), 4.15-4.11 (m, 2H), 3.93 (br s, 1H), 3.78-3.51 (m, 10H), 3.31-3.23 (m, 10H), 2.90 (t, J=7.5 Hz, 2H), 2.65-2.63 (m, 4H), 1.86-1.61 (m, 14H), 1.38-1.35 (m, 4H); ESI (m/z) [C.sub.49H.sub.79ClN.sub.12O.sub.13+H].sup.+ 1079; HPLC, AUC=98.4%; t.sub.R=8.26 min, Method A.

Example 83

Preparation of ethyl ((S)-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) amino)-1-((3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propyl)amino)-1-oxohexan-2-yl)carbamate (Exp-83)

##STR00452##

[0774] Preparation of Exp-83: In two steps Exp-83 was synthesized beginning with Int-29 and ethyl chloroformate using a sequence analogous to the one used to prepare Exp-78: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.28-7.24 (m, 4H), 4.12-4.07 (m, 5H), 3.83-3.63 (m, 10H), 3.37-3.34 (m, 5H), 3.24-3.22 (m, 5H), 2.74-2.65 (m, 4H), 1.97-1.67 (m, 10H), 1.47-1.44 (m, 2H), 1.24 (t, J=7.0 Hz, 3H); ESI (m/z) [C.sub.46H.sub.71ClN.sub.10O.sub.14+H].sup.+ 1023; HPLC, AUC=95.6%; t.sub.R=9.22 min, Method A.

Example 84

Preparation of N(N-(4-(4-(3-((S)-2-acetamido-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)hexanamido)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-3,5-diamino-6-chloropyrazine-2-carboxamide (Exp-84)

##STR00453##

[0775] Preparation of Exp-84: In two steps Exp-84 was synthesized beginning with Int-29 and acetic acid using a sequence analogous to the one used to prepare Exp-78: .sup.1H NMR (300 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.28-7.24 (m, 4H), 4.32-4.29 (m, 1H), 4.16-4.14 (m, 2H), 3.83-3.61 (m, 10H), 3.50-3.40 (m, 2H), 3.37-3.35 (m, 5H), 3.24-3.20 (m, 3H), 2.75-2.63 (m, 4H), 2.01 (s, 3H), 1.86-1.72 (m, 10H), 1.50-1.45 (m, 2H); ESI (m/z) [C.sub.45H.sub.69ClN.sub.10O.sub.13+H].sup.+ 993; HPLC, AUC=97.9%; t.sub.R=9.08 min, Method A.

Example 85

Preparation of 4-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)butyl 3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoate (Exp-85)

##STR00454##

[0776] Preparation of Exp-85a: To a solution of Int-23 (500 mg, 1.16 mmol) in DCM (10 mL) was added DCC (359 mg, 1.73 mmol) and DMAP (28.3 mg, 0.232 mmol) at room temperature. After the resultant mixture was stirred at room temperature for 10 min, N-Boc-4-amino-1-butanol (285 mg, 1.50 mmol) was added, and the resultant mixture was stirred at room temperature for 16 h. After the solvent was removed, the residue was purified by silica column (10% to 50% of EtOAc in CH.sub.2Cl.sub.2 followed by 10% to 50% of MeOH in CH.sub.2Cl.sub.2), to afford Exp-85a (557 mg, 80%) as a white solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 7.50-7.47 (m, 4H), 7.35-7.20 (m, 9H), 5.09 (s, 2H), 4.75 (br s, 1H), 4.60 (br s, 1H), 4.50 (br s, 1H), 3.70 (s, 3H), 4.10-4.08 (m, 1H), 3.68-3.67 (m, 1H), 3.48-3.47 (m, 1H), 3.24-2.96 (m, 6H), 2.70-2.60 (m, 4H), 1.70-1.50 (m, 6H), 1.45 (s, 9H), 1.40-1.30 (m, 2H); ESI (m/z) [C.sub.36H.sub.46N.sub.2O.sub.6+H].sup.+ 603.

[0777] Preparation of Exp-85b: Exp-85a (555 mg, 0.921 mmol) was dissolved in 4 N HCl in dioxane (1.5 mL). Solid precipitation was observed after 5 min. The resulting suspension was stirred at room temperature for 3 h. After the solvent was removed, MTBE (4.0 mL) was added. The solid was filtered out and washed with MTBE, to afford Exp-85b (373 mg, 75%) as a white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.50-7.47 (m, 4H), 7.33-7.22 (m, 9H), 5.05 (s, 2H), 4.11 (t, J=6.0 Hz, 2H), 3.13 (t, J=6.0 Hz, 2H), 2.96-2.91 (m, 4H), 2.69-2.66 (m, 4H), 1.83-1.33 (m, 8H); ESI (m/z) [C.sub.31H.sub.38N.sub.2O.sub.4+H].sup.+ 503.

[0778] Preparation of Exp-85c: To a solution of Exp-85b (50 mg, 0.093 mmol) in MeOH (2.0 mL) was added D-glucose (50.1 mg, 0.278 mmol) followed by AcOH (16.7 mg, 0.278 mmol) and NaCNBH.sub.3 (17.4 mg, 0.278 mmol). The resulting solution was stirred at 50 C. for 16 h. Additional D-glucose (33.5 mg, 0.186 mmol), AcOH (11.1 mg, 0.186 mmol) and NaCNBH.sub.3 (11.9 mg, 0.186 mmol) were added, and the reaction mixture was heated at 50 C. and stirred for 8 h. After the solvent was removed to the residue was added 1N HCl to adjust pH to 2 followed by stirring at room temperature for 3 min. The solution was purified by reverse phase column (5% to 100% of CH.sub.3CN in H.sub.2O, product came out at 50% to 80%), to afford Exp-85c (51 mg, 63%) as a colorless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.50-7.47 (m, 4H), 7.32-7.22 (m, 9H), 5.05 (s, 2H), 4.08-3.34 (m, 18H), 3.16-2.94 (m, 6H), 2.68-2.64 (m, 4H), 1.72-1.53 (m, 8H); ESI (m/z) [C.sub.43H.sub.62N.sub.2O.sub.14+H].sup.+ 831.

[0779] Preparation of Exp-85d: To a solution of Exp-85c (192 mg, 0.221 mmol) in EtOH (5.0 mL) and water (5.0 mL) was added 20% Pd(OH).sub.2 (20 mg). The resultant mixture was stirred under hydrogen (balloon) at room temperature for 4 h. No conversion was observed. After filtration, fresh 20% Pd(OH).sub.2 (20 mg) was added, and the resultant suspension was stirred under hydrogen balloon for 16 h. Around 20% conversion was observed. After filtration, fresh 20% Pd(OH).sub.2 (20 mg) was added, and the resultant suspension was stirred under hydrogen balloon at 45 C. for 6 h. After filtration, the solvent was removed, to afford Exp-85d (132 mg, 81%) as white syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.29-7.26 (m, 4H), 4.09-4.08 (m, 2H), 3.78-3.59 (m, 10H), 3.50-3.40 (m, 4H), 2.97-2.91 (m, 6H), 2.73-2.54 (m, 4H), 1.77-1.63 (m, 8H); ESI (m/z) [C.sub.35H.sub.56N.sub.2O.sub.12+H].sup.+ 697.

[0780] Preparation of Exp-85: Exp-85d (85.0 mg, 0.116 mmol) was suspended in MeOH (5.0 ml), and 3 N HCl in MeOH (2.0 equiv) was added. The resultant mixture was stirred at room temperature for 5 min, to give a clear solution. The solvent was removed, and the residue was azeotrop with MeOH (3.0 mL2). The residue was dissolved in DMF (2.0 mL) and a clear solution observed. Compound 1 (45.0 mg, 0.116 mmol) and DIPEA (90.0 mg, 0.695 mmol) was added. Solid precipitation was observed. The resulting suspension was stirred at 65 C. for 8 h. After concentration, 1 N HCl (3.0 mL) was added to the residue and stirred for 3 min. The solution was purified by reverse phase column (5% to 95% of CH.sub.3CN in H.sub.2O, product came out at 40%), to afford Exp-85 (46 mg, 40%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.29-7.27 (m, 4H), 4.15-4.13 (m, 4H), 3.82-3.64 (m, 10H), 3.50-3.40 (m, 4H), 3.39-3.30 (m, 4H), 2.96 (t, J=8.0 Hz, 2H), 2.73-2.68 (m, 4H), 1.79-1.74 (m, 8H); ESI (m/z) [C.sub.41H.sub.61ClN.sub.8O.sub.13+H].sup.+ 909; HPLC, AUC=96.8%; t.sub.R=9.26 min, Method A.

Example 86

Preparation of 3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino) butyl)-[1,1-biphenyl]-4-yl)propyl 3-(((2R,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) ((2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)propanoate (Exp-86)

##STR00455##

[0781] Preparation of Exp-86a: To a stirred solution of compound Int-16 (2.0 g, 13.4 mmol) in dichloromethane (50 mL) was added DCC (3.20 g, 15.4 mmol) and DMAP (250 mg, 2.06 mmol) at room temperature. The reaction mixture was stirred for 15 minutes, then 3-((tert-butoxycarbonyl)amino)propanoic acid (4.30 g, 10.3 mmol) was added and stirring continued for 16 h at room temperature. After completion of the reaction, the reaction mass was filtered, the solids were washed with MTBE (25 mL) and the filtrate was evaporated under reduced pressure to get crude material. The obtained crude was purified by combi-flash (40 g column) chromatography eluted with 5% EtOAc in hexanes to afford compound Exp-86a (2.60 g, 43%) as an off white solid. ESI (m/z) [C.sub.35H.sub.44N.sub.2O.sub.6+H]+589.

[0782] Preparation of Exp-86b: To a stirred solution of Exp-86a (2.60 g, 57.6 mmol) in triflouroethanol (15 mL) was added TMSCl (6 mL) at 0 C. The reaction mixture was stirred for 1 h at room temperature. After completion of the reaction, the reaction was concentrated under reduced pressure to afford Exp-86b (1.20 g, crude) as an off-white solid. ESI (m/z) [C.sub.30H.sub.36N.sub.2O.sub.4+H].sup.+ 489.

[0783] Preparation of Exp-86c: To a stirred solution of Exp-86b (1.20 g, 2.46 mmol) in MeOH was added D-Glucose (1.3 g, 7.39 mmol), NaCNBH.sub.3 (0.46 g, 7.39 mmol) and acetic acid (0.45 g, 7.39 mmol) at room temperature. The reaction mixture was stirred for 16 h at 60 C. After completion of the reaction, the reaction mass was evaporated under reduced pressure to get crude material. The obtained crude was purified by reverse phase combi flash (220 g column) chromatography eluted with 30% ACN:Water (0.1% HCl) to afford Exp-86c (0.70 g, 35%) as an off-white solid. ESI (m/z) [C.sub.42H.sub.60N.sub.2O.sub.14+H].sup.+ 817.

[0784] Synthesis of Exp-86d: To a stirred solution of Exp-86c (0.70 g 0.85 mmol) in IPA:water (7:3) was added catalytic amount of acetic acid and Pd(OH).sub.2 (250 mg) at room temperature. The reaction mixture was stirred at room temperature under H.sub.2 (balloon) for 1 h. After completion of the reaction, the reaction mass was filtered through celite, the solids were washed with IPA:water (20 mL, 1:1) and filtrate was concentrated to get crude material. The obtained crude was purified by reverse phase combi flash (220 g column) chromatography eluting with 23% ACN:water (0.1 M HCl) to afford Exp-86d (0.20 g, 31%) as an off-white solid. ESI (m/z) [C.sub.34H.sub.54N.sub.2O.sub.12+H].sup.+ 683.

[0785] Preparation of Exp-86: To a stirred solution of Exp-86d (0.2 g, 0.29 mmol) in DMF (4 mL), was added DIPEA (0.25 mL, 1.45 mmol) and compound 1 (76 mg, 0.29 mmol) at room temperature. The reaction mixture was warmed to 55 C. and stirred for 5 h. After completion of reaction, the reaction mass was directly evaporated under reduced pressure to get crude material. The obtained crude was purified by reverse phase combi flash (220 g column) chromatography eluted with 16% ACN:water (0.1 M HCl) to afford Exp-86 (0.053 g, 22%) as an off-white solid. .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.46-7.39 (m, 4H), 7.21-7.14 (m, 4H), 4.15-4.053 (m, 4H), 3.76-3.65 (m, 5H), 3.61-3.52 (m, 6H), 3.46-3.24 (m, 7H), 2.91-2.84 (m, 2H), 2.67-2.61 (m, 4H), 1.95-1.87 (m, 2H), 1.72-1.61 (m, 4H)); ESI (m/z) [C.sub.40H.sub.59ClN.sub.8O.sub.13+H].sup.+ 895; HPLC, AUC=95.6%; t.sub.R=6.32 min, Method J.

Example 87

Preparation of 3,5-diamino-N(N-(4-(4-(3-((2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-87)

##STR00456##

[0786] Preparation of Exp-87a: Int-23 (90 mg, 0.209 mmol), Int-20 (89 mg, 0.209 mmol) and DIPEA (81 mg, 0.626 mmol) were dissolved in DMF (3.0 mL), stirred at rt for 10 min then HATU (95 mg, 0.250 mmol) was added. The resulting solution was stirred at rt for 2 h. Solvent was removed under reduced pressure. The residue was dissolved in water and pH was adjusted to 3. The acidic solution was purified by reverse phase column (5-80% CH.sub.3CN in H.sub.2O, product came out at 40%), to afford Exp-87a (126 mg, 72%) as colorless syrup. .sup.1H NMR (500 MHz, CD.sub.3OD) 7.51-7.48 (m, 4H), 7.33-7.22 (m, 9H), 5.06 (s, 2H), 4.20-4.18 (m, 2H), 3.83-3.34 (m, 18H), 3.14 (t, J=6.0 Hz, 2H), 3.00-2.96 (m, 2H), 2.65-2.57 (m, 4H), 1.70-1.50 (m, 4H).

[0787] Preparation of Compound Exp-87b: To a solution of Exp-87a (123 mg, 0.147 mmol) in MeOH (5.0 mL) was added Pd(OH).sub.2/C (50 mg). The suspension was stirred under H.sub.2 (balloon) for 6 h. After filtration, the filtrate was concentrated to afford Exp-87b (98 mg, 95%) as colorless syrup. .sup.1H NMR (500 MHz, CD.sub.3OD) Q 7.53-7.51 (m, 4H), 7.29-7.26 (m, 4H), 4.22-4.18 (m, 2H), 3.84-3.34 (m, 18H), 2.96-2.94 (m, 4H), 2.75-2.57 (m, 4H), 1.80-1.70 (m, 4H).

[0788] Preparation of Exp-87: To a solution of Exp-87b (615 mg, 0.873 mmol) in EtOH (10 mL) and DMF (10 mL) was added DIPEA (677 mg, 5.24 mmol) and compound 1 (373 mg, 0.961 mmol). The resulting solution was stirred at 70 C. for 8 h. The solvent was removed under reduced pressure and EtOH (10 mL) was added resulting in precipitation. The solid was collected, suspended in EtOH/H.sub.2O (20 mL/20 mL) and warmed to 50 C. resulting in a clear solution. The volume of solution was reduced to around 8 mL resulting in precipitation. Additional EtOH (50 mL) was added to complete the precipitation, the solid was collected by filtration and dried to afford Exp-87 (578 mg, 72%) as yellow-brown solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.50 (m, 4H), 7.28-7.27 (m, 4H), 4.19 (br s, 2H), 3.84-3.34 (m, 20H), 2.98 (t, J=6.0 Hz, 2H), 2.73 (t, J=6.0 Hz, 2H), 2.57 (t, J=6.0 Hz, 2H), 1.79-1.75 (m, 4H). ESI-MS m/z [C.sub.39H.sub.58ClN.sub.9O.sub.12+H].sup.+ 880; HPLC, AUC=98.4%; t.sub.R=8.97 min, Method A.

Examples 88-97

[0789] Preparation of Examples 88-97: In a scheme analogous to the one used for preparation of Example 87, the intermediates designated below were converted to corresponding Examples 88-97. In examples with unsaturation in the side chain it was reduced when the Cbz protecting group was removed. In examples with a Boc protecting group, it was removed with TFA/C.sub.2H.sub.2 or HCl/MeOH:

TABLE-US-00036 Exp-# Int-# Structure Spectra 88 Int-8 [00457] .sup.1H NMR (500 MHz, D.sub.2O) 7.71 (d, J = 8.5 Hz, 2H), 7.58-7.56 (m, 4H), 7.31 (d, J = 8.0 Hz, 2H), 4.21 (br s, 2H), 3.87-3.40 (m, 18H), 3.13 (t, J = 6.0 Hz, 2H), 2.61 (t, J = 6.0 Hz, 2H), 1.66-1.63 (m, 4H); ESI (m/z) [C.sub.37H.sub.54ClN.sub.9O.sub.12 + H].sup.+ 852; HPLC, AUC = 99.1%; t.sub.R = 7.89 min, Method A. Exp-88 89 Int-9 [00458] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.56-7.52 (m, 4H), 7.36-7.28 (m, 4H), 4.13 (br s, 2H), 3.81-3.59 (m, 14H), 3.50-3.40 (m, 2H), 3.35-3.34 (m, 5H), 2.73 (t, J = 7.0 Hz, 3H), 1.81-1.72 (m, 4H); ESI (m/z) [C.sub.38H.sub.56ClN.sub.9O.sub.12 + H].sup.+ 866; HPLC, AUC = 94.7%; t.sub.R = 8.73 min, Method A. Exp-89 90 Int-11 [00459] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.28-7.27 (m, 4H), 4.18-4.13 (m, 2H), 3.81- 3.34 (m, 20H), 2.74-2.66 (m, 4H), 2.30-2.28 (m, 2H), 2.02-1.93 (m, 2H), 1.79-1.73 (m, 4H); ESI (m/z) [C.sub.40H.sub.60ClN.sub.9O.sub.12 + H].sup.+ 894; HPLC, AUC = 96.6%; t.sub.R = 9.01 min, Method A. Exp-90 91 Int-12 [00460] .sup.1H NMR(400 MHz, MeOH-d.sub.4) 7.46-7.42 (d, 2H), 7.36-7.31(d, 2H), 7.25-7.18 (m, 3H) 7.10- 7.07 (d, 1H), 4.12-4.06 (m, 2H), 3.75-3.75 (m, 2H), 3.70-3.67 (m, 2H), 3.61-3.54 (m, 6H), 3.48- 3.41 (m, 4H), 3.37-3.33 (m, 3H), 3.29-3.266 (m, 3H), 2.92-2.87 (m, 2H), 2.66-2.62 (m, 2H) 2.52- 2.47 (m, 2H), 1.74-1.65 (m, 4H).); [. ESI (m/z) [C.sub.39H.sub.58ClN.sub.9O.sub.12 + H].sup.+ 880; HPLC, AUC = 99.3%; t.sub.R = 6.21 min, Method I. Exp-91 92 Int-13 [00461] .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.16 (d, J = 8.0 Hz, 2H), 7.10 (d, J = 16 Hz, 2H), 7.05 (s, 1H), 6.91(d, 2H), 3.80-3.75(m, 2H), 3.70-3.66(M, 4H), 3.63-3.58 (m, 2H)), 3.56- 3.50(m, 4H), 3.28-3.23 (m, 3H), 3.18-3.12 (m, 1H), 2.83-2.77 (t, 2H), 2.67-2.52 (m, 8H), 2.44- 2.41 (t, 2H), 2.11 (s, 3H) 1.85-1.81 (m, 5H), 1.72- 1.62 (m, 4H); ESI (m/z) [C.sub.40H.sub.60ClN.sub.9O.sub.12+ H].sup.+ 894; HPLC, AUC = 96.5%; t.sub.R = 6.78 min, Method G. Exp-92 93 Int-14 [00462] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.49-7.42 (m, 4H), 7.26-7.21 (m, 4H), 3.81-3.65 (m, 11H), 3.50- 3.40 (m, 11H), 3.00-2.93 (m, 2H), 2.73-2.68 (m, 2H), 2.58-2.52 (m, 2H), 1.77-1.69 (m, 4H), 1.45- 1.42 (m, 2H); ESI (m/z) [C.sub.40H.sub.60ClN.sub.9O.sub.12 + H].sup.+ 894; UPLC: 98.1% AUC, t.sub.R = 2.78 min, Method B. Exp-93 94 Int-15 [00463] .sup.1H NMR (500 MHz, CD.sub.3OD) 7.49-7.42 (m, 4H), 7.26-7.21 (m, 4H), 3.81-3.65 (m, 11H), 3.50- 3.40 (m, 11H), 3.00-2.93 (m, 2H), 2.73-2.68 (m, 2H), 2.58-2.52 (m, 2H), 1.77-1.69 (m, 4H), 1.45- 1.42 (m, 2H); ESI (m/z) [C.sub.40H.sub.60ClN.sub.9O.sub.12 + H].sup.+ 894; UPLC: 98.1% AUC, t.sub.R = 2.78 min, Method B. Exp-94 95 Int-19 [00464] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.30-7.40 (m, 4H), 7.13 (d, J = 8.0 Hz, 2H), 6.78 (d, J = 8.0 Hz, 2H), 4.47 (s, 2H), 4.10 (m, 2H), 3.75-3.60 (m, 8H), 3.56-3.42 (m, 6H), 3.38- 3.24 (m, 7H), 2.96 (m, 2H), 2.48 (m, 2H), 1.60- 1.40 (m, 4H); ESI (m/z) [C.sub.38H.sub.56ClN.sub.9O.sub.13 + H].sup.+ 882; HPLC, AUC = 99.8%; t.sub.R = 6.19 min, Method J. Exp-95 96 Int-52 [00465] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.42 (d, J = 8.0 Hz, 2H), 7.35-7.31 (m, 2H), 7.26-7.22 (m, 1H), 7.17 (d, J = 8.0 Hz, 2H), 7.09 (d, J = 7.6 Hz, 1H), 4.08-4.07 (m, 2H), 3.74-3.67 (m, 4H), 3.63- 3.53 (m, 6H) 3.47-3.3.29 (m, 10H), 2.85 (t, J = 8 Hz, 2H), 2.67 (t, J = 6.8 Hz, 2H), 2.47 (t, J = 8 Hz, 2H), 1.72-1.64(m, 4H); ESI (m/z) [C.sub.36H.sub.55ClN.sub.10O.sub.12S + H].sup.+ 880; HPLC, AUC = 97.52%; t.sub.R = 6.25 min, Method I. Exp-96 97 Int-47 [00466] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.26-7.11 (m, 7H), 4.00-3.95 (m, 2H), 3.72-3.63 (m, 4H), 3.61- 3.52 (m, 6H), 3.42-3.41 (m, 2H), 3.28-3.09 (m, 8H), 2.86 (t, J = 7.6 Hz, 2H), 2.65 (t, J = 7.2 Hz, 2H), 2.47 (t, J = 7.6 Hz, 2H), 1.73-1.58 (m, 4H); ESI (m/z) [C.sub.39H.sub.57Cl.sub.2N.sub.9O.sub.12 + H].sup.+ 914; HPLC, AUC = 96.1%; tR = 6.23 min, Method J. Exp-97

Alternative Synthesis of Example 92

Preparation of 3,5-diamino-N(N-(4-(4-(3-((2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)amino)-3-oxopropyl)-2-methyl-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-92)

##STR00467##

[0790] Preparation of Exp-92a: To a stirred solution of Int-13 (2.0 g, 6.5 mmol) in THF:water (6:4, 50 mL) was added LiOH (414 mg, 9.8 mmol) followed by stirring for 16 h at room temperature. After completion of the reaction, the reaction mass was acidified with 2N HCl and extracted with EtOAc (350 mL). The combined organic layer was washed with water (50 mL), brine solution (50 mL) and dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to get desire Exp-92a (1.80 g, 63%) as an off white solid. ESI (m/z) [C.sub.28H.sub.31NO.sub.4+H].sup.+ 446.

[0791] Preparation of Exp-92b: To a stirred solution of Exp-92a (0.80 g, 2.5 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added TEA (1.30 mL, 8.98 mmol) and T.sub.3P (3.50 g, 11.2 mmol). The reaction mixture was stirred for 10 min, tert-butyl (2-aminoethyl)carbamate was added and stirring continued for 16 h at room temperature. The reaction mass was quenched with NaHCO.sub.3 solution (100 mL) and extracted with CH.sub.2Cl.sub.2 (350 mL). The combined organic extracts were concentrated under reduced pressure to get crude material. The obtained crude was purified by combi-flash (40 g column) chromatography eluted with 10% MeOH:dichloromethane to afford Exp-92b (1.60 g, 60%) as an off white solid. ESI (m/z) [C.sub.35H.sub.45N.sub.3O.sub.5+H].sup.+ 588.

[0792] Preparation of Exp-92c: To a stirred solution of Exp-92b (0.80 g, 2.5 mmol) in dichloromethane (20 mL) was added 4M HCl (3 mL) at 0 C. followed by stirring for 3 h at room temperature. After completion of the reaction, the reaction mass was concentrated under reduced pressure to afford Exp-92c (1.5 g, crude) as an off-white solid. ESI (m/z) [C.sub.30H.sub.37N.sub.3O.sub.3+H].sup.+ 488.

[0793] Preparation of Exp-92d: To a stirred solution of Exp-92c (1.60 g, 3.2 mmol) in MeOH (15 mL) was added D-Glucose (1.80 g, 9.8 mmol), NaCNBH.sub.3 (0.60 g, 9.8 mmol) and acetic acid (0.59 mg, 9.8 mmol) at room temperature. The reaction mixture was warmed to 60 C. and stirred for 16 h. After completion of the reaction, the reaction mass was concentrated under reduced pressure to get crude material. The obtained crude was purified by reverse phase combiflash (220 g column) chromatography eluted with 50% ACN:water (0.1% HCl) to afford compound Exp-92d (1.50 g, 57%) as an off-white solid. ESI (m/z) [C.sub.42H.sub.61N.sub.3O.sub.13+H].sup.+ 816.

[0794] Preparation of Exp-92e: To a stirred solution of Exp-92d (1.6 g 1.96 mmol) in IPA:water (7:3, 20 mL) was added a catalytic amount of AcOH and Pd(OH).sub.2 (500 mg). The reaction mixture was stirred under H.sub.2 (balloon) for 16 h at room temperature. After completion of the reaction, the reaction mass was filtered through celite and the solids were washed with IPA:water (75 mL). The filtrate was concentrated under reduced pressure to get crude material. The obtained crude was purified by reverse phase combiflash (220 g column) chromatography eluting with 23% ACN:water (0.1% AcOH) to afford Exp-92e (1.10 g, 82%) as an off-white solid. ESI (m/z) [C.sub.34H.sub.55N.sub.3O.sub.11+H].sup.+ 682.

[0795] Preparation of Exp-92: To a stirred solution of Exp-92e (0.50 g, 0.73 mmol) in DMF (5 mL) was added DIPEA (0.7 mL, 4.4 mmol) and 1 (191 mg, 0.73 mmol) at room temperature. The reaction mixture was warmed to 55 C. and stirred for 5 h. After completion of the reaction, reaction mass was concentrated under reduced pressure to get crude material. The obtained crude was purified by reverse phase combi flash (220 g column) chromatography eluting with 20% ACN:water (0.1 M AcOH) to afford Exp-92 (0.22 g, 33%) as an off-white solid. .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.16 (d, J=8.0 Hz, 2H), 7.10 (d, J=16 Hz, 2H), 7.05 (s, 1H), 6.91 (d, 2H), 3.80-3.75 (m, 2H), 3.70-3.66 (m, 4H), 3.63-3.58 (m, 2H), 3.56-3.50 (m, 4H), 3.28-3.23 (m, 3H), 3.18-3.12 (m, 1H), 2.83-2.77 (t, 2H), 2.67-2.52 (m, 8H), 2.44-2.41 (t, 2H), 2.11 (s, 3H), 1.85-1.81 (m, 5H), 1.72-1.62 (m, 4H); ESI (m/z) [C.sub.40H.sub.60ClN.sub.9O.sub.12+H].sup.+ 894; HPLC, AUC=96.5%; t.sub.R=6.78 min, Method G.

Example 98

Preparation of 3,5-diamino-N(N-(4-(4-(3-((3-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)propyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-98)

##STR00468##

[0796] Preparation of Exp-98: In a three-step procedure Exp-98 was synthesized beginning with Int-10 and Int-21 using a sequence analogous to the one used to prepare Exp-87: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.29-7.27 (m, 4H), 4.07 (br s, 2H), 3.81-3.62 (m, 10H), 3.50-3.34 (m, 10H), 2.95 (d, J=7.5 Hz, 2H), 2.73 (d, J=7.0 Hz, 2H), 2.54 (d, J=7.5 Hz, 2H), 1.85-1.72 (m, 6H); ESI (m/z) [C.sub.40H.sub.60ClN.sub.9O.sub.12+H].sup.+ 894; HPLC, AUC=97.5%; t.sub.R=8.46 min, Method A.

Example 99

Preparation of 3,5-diamino-N(N-(4-(4-(3-((4-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)butyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-99)

##STR00469##

[0797] Preparation of Exp-99: In a five-step procedure Exp-99 was synthesized beginning with Int-23 and tert-butyl (2-aminobutyl)carbamate using a sequence analogous to the one used in the alternate synthesis of Exp-92: .sup.1H NMR (500 MHz, D.sub.2O) 7.53-7.50 (m, 4H), 7.29-7.27 (m, 4H), 4.18-4.14 (m, 2H), 3.82-3.64 (m, 10H), 3.50-3.40 (m, 2H), 3.38-3.20 (m, 8H), 2.95 (d, J=7.5 Hz, 2H), 2.73 (d, J=7.0 Hz, 2H), 2.53 (d, J=7.5 Hz, 2H), 1.79-1.72 (m, 6H), 1.57-1.54 (m, 2H); ESI (m/z) [C.sub.41H.sub.62ClN.sub.9O.sub.12+H].sup.+ 908; HPLC, AUC=95.2%; t.sub.R=8.53 min, Method A.

Example 100

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(3-(((6S,7S,8R,9R,10R)-2-((4S,5S,6R,7R,8R)-4,5,6,7,8,9-hexahydroxynonyl)-6,7,8,9,10,11-hexahydroxy undecyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-100)

##STR00470##

[0798] Preparation of Exp-100a: To a stirred solution of compound Int-43 (0.20 g, 0.46 mmol in dichloromethane (10 mL) was added TEA (0.10 mL, 0.92 mmol), compound Int-23 (0.49 g, 0.69 mmol) and HATU (0.26 g, 0.69 mmol) at 0 C. The reaction mixture was warmed to room temperature and stirred for 2 h. After completion of the reaction, the reaction mixture was evaporated under reduced pressure to get crude material which was purified by combi flash (120 g column) chromatography eluting with 50% EtOAc in hexanes to afford compound Exp-100a (0.50 g, 95%) as an off-white solid. ESI (m/z) [C.sub.65H.sub.94N.sub.2O.sub.15+H].sup.+ 1143.

[0799] Preparation of Exp-100b: To a stirred solution of Exp-100a (0.50 g 0.43 mmol) in a mixture of IPA:water (7:3) was added a catalytic amount of acetic acid and 20% Pd(OH).sub.2 on carbon 50% wet (100 mg, 50% weight substrate) at room temperature. The reaction mixture was stirred under H.sub.2 (balloon) at room temperature for 16 h. After completion of the reaction, the reaction mass was filtered through celite and the solids were washed with (1:1) IPA:H.sub.2O (20 mL). The combined filtrate was evaporated under reduced pressure to afford Exp-100b (0.20 g, 45%) as an off white solid. ESI (m/z) [C.sub.57H.sub.88N.sub.2O.sub.13+H].sup.+ 1009.

[0800] Preparation of Exp-100c: To a stirred solution of Exp-100b (0.20 g, 0.19 mmol) in DMF (3 mL) was added DIPEA (0.2 mL, 1.14 mmol) and compound 1 (0.052 g, 0.19 mmol). The reaction mixture was warmed to 60 C. and stirred for 6 h. After completion of reaction, the reaction mass was directly evaporated under reduced pressure to get crude which was purified by reverse phase combi flash (220 g column) chromatography eluting with 33% ACN:water (0.1 M HCl) to afford Exp-100c (0.09 g, 46%). ESI (m/z) C63H93ClN8O14+H].sup.+ 1221.

[0801] Preparation of Exp-100: To a stirred solution of compound Exp-100c (0.09 g, 0.08 mmol) in MeOH:water (2:1) was added 4M HCl in methanol (1 mL). The reaction mixture was warmed to 60 C. and stirred for 2 h. After completion of the reaction, the reaction mass was directly evaporated under reduced pressure to afford desired compound Exp-100 (53 mg, 60%) as an off-white solid. .sup.1H NMR (400 MHz, CD.sub.3OD) 7.34-7.19 (m, 4H), 7.96-6.95 (m, 4H), 3.86-3.82 (m, 2H), 3.71-3.63 (m, 5H), 3.55-3.51 (m, 4H), 3.48-3.45 (m, 2H), 3.42-3.38 (m, 2H), 3.08-3.02 (m, 2H), 2.88-2.84 (m, 2H), 2.75-2.72 (m, 2H), 2.48-2.39 (m, 3H), 1.57-1.46 (m, 4H), 1.40-1.41 (m, 3H), 1.20-1.16 (m, 4H), 1.08-1.02 (m, 2H), 0.93-0.81 (m, 4H); ESI (m/z) [C.sub.45H.sub.69ClN.sub.8O.sub.14+H].sup.+ 981; HPLC, AUC=93.1%; t.sub.R=6.23 min, Method K.

Examples 101-103

[0802] Preparation of Examples 101-103: In a scheme analogous to the four-step procedure used for preparation of Example 100, intermediates Int-31, 32 and 33 were coupled to Int-23 and moved forward to Examples 101-103. However, for Examples 100 and 101 the acetonides were removed in the second step and third step (10% H.sub.2SO.sub.4) respectively.

TABLE-US-00037 Exp-# Structure Spectra 101 [00471] .sup.1H NMR (400 MHz, DMSO- d6) 7.57-7.55 (m, 4H), 7.32- 7.27 (m, 4H), 6.84 (s, 2H), 6.61 (s, 1H), 4.96 (s, 1H), 4.71-4.56 (m, 5H), 4.36-4.23 (m, 3H), 4.09-4.07 (m, 2H), 3.95-3.91 (m, 2H), 3.87-3.83 (m, 4H), 3.59-3.46 (m, 9H), 3.15-3.13 (m, 2H), 2.95-2.91 (m, 2H), 2.67-2.60 (m, 6H), 1.86-1.84 (m, 3H), 1.64-1.53 (m, 4H); ESI (m/z) [C.sub.45H.sub.61ClN.sub.8O.sub.16 + H].sup.+ 1005; HPLC, AUC = 93.5%; t.sub.R = 6.29 min, Method I. Exp-101 102 [00472] .sup.1H NMR (400 MHz, DMSO- d.sub.6) 8.34 (t, J = 6.0, 1H), 7.55-7.53 (d, J = 8.0 Hz, 4H), 7.29-7.27 (d, J = 7.2 Hz, 4H), 6.87-6.61 (m, 2H), 6.42-6.41 (d, J = 1.6 Hz, 2H), 6.39-6.22 (m, 1H), 4.93 (d, J = 5.2 Hz, 2H), 4.77-4.63 (m, 2H), 4.60- 4.54 (m, 3H), 4.43-4.29 (m, 2H), 4.2 (d, J = 7.2 Hz, 3H), 4.12 (d, J = 8.4 Hz, 2H), 3.99-3.92 (m, 2H), 3.91-3.83 (m, 4H), 3.59-3.46 (m, 8H), 3.41-3.37 (m, 2H), 3.21-3.15 (m, 2H), 2.87 (t, J = 7.2 Hz, 2H), 2.67-2.64 (m, 3H), 2.33- 2.32 (m, 2H), 1.90 (s, 5H), 1.65-1.63 (m, 2H), 1.56-1.51 (m, 2H); ESI (m/z) [C.sub.46H.sub.63ClN.sub.8O.sub.16 + H].sup.+ 1019; HPLC, AUC = 98.1%; t.sub.R = 6.27 min, Method J. Exp-101 103 [00473] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.38-7.33 (m, 4H), 7.18- 7.14 (m, 4H), 6.304 (s, 1H), 6.28 (d, J = 1.72 Hz, 2H), 4.10-4.08 (m, 4H), 3.97-3.91 (m, 4H), 3.75-3.68 (m, 3H), 3.67-3.62 (m, 5H), 3.58-3.56 (m, 2H), 3.55-3.53 (m, 2H), 3.04 (t, J = 6.59 Hz, 2H), 2.84 (t, J = 7.4 Hz, 2H), 2.62 (t, J = 7.19 Hz, 2H), 2.41 (t, J = 7.68, 2H), 2.34 (t, J = 7.5 Hz, 2H), 1.71-1.56 (m, 6H); ESI (m/z) [C.sub.48H.sub.67ClN.sub.8O.sub.16 + H].sup.+ 1047; HPLC, AUC = 98.4%; t.sub.R = 6.29 min, Method I.

Example 104

Preparation of 3,5-diamino-N(N-(4-(4-(3-((2-(bis(3-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)propyl)amino)ethyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-104)

##STR00474## ##STR00475##

[0803] Preparation of Exp-104a: To a stirred solution of compound Int-34 (850 mg, 1.669 mmol) and tert-butyl (3-oxopropyl)carbamate (866 mg, 5.009 mmol) in MeOH (20 mL), AcOH (300 mg, 5.009 mmol) and NaBH.sub.3CN (310 mg, 5.009 mmol) were added. The reaction mixture stirred at room temperature for 36 h. The reaction mass was charged with another lot of tert-butyl (3-oxopropyl)carbamate (866 mg, 5.009 mmol) and NaBH.sub.3CN (310 mg, 5.009 mmol), then stirring continued for another 24 h. The reaction mass concentrated under reduced pressure and the obtained crude was portioned between saturated NaHCO.sub.3 (50 mL) and EtOAc (50 mL). The separated EtOAc layer was washed with brine solution (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The crude was purified by combi-flash column chromatography eluting with 1%-10% MeOH in dichloromethane (compound isolated at 2% MeOH) to afford compound Exp-104a (700 mg, 53%) as a gummy liquid: ESI (m/z) [C.sub.45H.sub.65N.sub.5O.sub.7+H].sup.+ 788.

[0804] Preparation of Exp-104b: To a solution of compound Exp-104a (700 mg, 0.888 mmol) in dichloromethane (10 mL), TFA (1.0 mL) was added slowly followed by stirring at room temperature for 2 h. The reaction mass concentrated under reduced pressure and obtained crude azeotroped with dichloromethane (320 mL) to afford Exp-104b (800 mg, 97%) as a light brown liquid: ESI (m/z) [C.sub.35H.sub.49N.sub.5O.sub.3+H].sup.+ 588.

[0805] Preparation of Exp-104c: To a solution of Exp-104b (830 mg, 0.893 mmol) and D(+)-glucose (964 mg, 5.36 mmol) in MeOH (30 mL) was added NaBH.sub.3CN (288 mg, 4.59 mmol). The resultant reaction mixture was heated to 60 C. and stirred for 24 h. The reaction mass was cooled to room temperature and charged with D(+)-glucose (482 mg, 2.68 mmol) followed by NaBH.sub.3CN (144 mg, 2.29 mmol) then continued stirring at 60 C. for another 24 h. The reaction mass concentrated under reduced pressure. The obtained crude was treated with 3N HCl in CH.sub.3OH (10 mL) for 15 min and concentrated under reduced pressure. The crude was purified by reverse phase combi flash chromatography eluting with 10%-100% CH.sub.3CN in H.sub.2O (compound isolated at 40% of CH.sub.3CN) to afford Exp-104c (450 mg, 40%) as a gummy solid: ESI (m/z) [C.sub.59H.sub.97N.sub.5O.sub.23+H].sup.+ 1244.

[0806] Preparation of Exp-104d: To a solution Exp-104c (450 mg, 0.36 mmol) in IPA (20 mL) and H.sub.2O (20 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (100 mg, 50% weight substrate) under inert atmosphere. The resultant reaction mixture stirred under H.sub.2 (balloon) for 16 h. The reaction mass filtered through celite bed and the filtrate was concentrated under reduced pressure. The obtained crude was azeotroped with CH.sub.3CN (220 mL) and dried under reduced pressure to afford Exp-104d (400 mg, 70%) as a gummy liquid: ESI (m/z) [C.sub.51H.sub.91N.sub.5O.sub.21+H].sup.+ 1110.

[0807] Preparation of Exp-104: To a solution of Exp-104d (300 mg, 0.246 mmol) and compound 1 (105 mg, 0.27 mmol) in DMF (10 mL) was added DIPEA (0.2 mL, 1.23 mmol). The reaction mixture was heated to 60 C. and stirred for 16 h. The reaction mixture was concentrated under reduced pressure and the obtained crude was dissolved in water (5 mL) and the pH was adjusted to 2 with 2N HCl. The resulting solution was purified by reverse phase combiflash column chromatography eluting with 10%-100% of ACN in H.sub.2O (0.01% Conc. HCl) (compound isolated at 30% of ACN) to afford Exp-104 (120 mg, 33%) as an off-white solid: .sup.1H NMR (400 MHz, D.sub.2O) 7.55 (d, J=7.6 Hz, 2H), 7.43 (d, J=7.6 Hz, 2H), 7.30 (d, J=8.0 Hz, 2H), 7.16 (d, J=7.6 Hz, 2H), 4.16-4.08 (m, 4H), 3.76-3.65 (m, 12H), 3.57-3.52 (m, 9H), 3.42-3.35 (m, 3H), 3.32-3.25 (m, 10H), 3.18-3.08 (m, 8H), 2.85-2.81 (m, 2H), 2.58-2.53 (m, 4H), 2.12-2.08 (m, 4H), 1.75-1.68 (m, 2H), 1.64-0.8 (m, 2H); ESI (m/z) [C.sub.57H.sub.96ClN.sub.11O.sub.22+H].sup.+ 1322; HPLC, AUC=99.1%; t.sub.R=7.55 min, Method X.

Example 105

Preparation of methyl N2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N6-((2S,3R,4R,5R)-2,3,4,5,6-penta hydroxyhexyl)-L-lysinate (Exp-105)

##STR00476##

[0808] Preparation of Exp105a: To a solution of Int-35 (100 mg, 0.164 mmol) in MeOH (5.0 mL) was added D-glucose (32.5 mg, 0.180 mmol) followed by NaCNBH.sub.3 (15.4 mg, 0.246 mmol). The resulting solution was stirred at 50 C. for 6 h. The solvent was removed, and the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in H.sub.2O, product came out at 50%). The pure fractions were collected, and solvents were removed. The residue was azeotrope with MeOH to remove residual water, then 3N HCl in MeOH (0.55 mL) was added, and azeotrope with MeOH again to remove borane, to afford Exp-105a (72 mg, 56%) as a white solid: ESI (m/z) [C.sub.40H.sub.55N.sub.3O.sub.10+H].sup.+ 738.

[0809] Preparation of Exp-105b: To a solution of Exp-105a (72 mg, 0.093 mmol) in EtOH (2.0 mL) and water (2.0 mL) was added 20% Pd(OH).sub.2 (15 mg) and AcOH (33.5 mg, 0.558 mmol). The resultant mixture was stirred under hydrogen (balloon) at 40 C. for 6 h. After filtration, the solvent was removed, to afford Exp-105b (61 mg, 91%) as colorless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.29-7.27 (m, 4H), 4.44-4.41 (m, 1H), 4.02-4.01 (m, 1H), 3.83-3.43 (m, 8H), 3.11-2.91 (m, 8H), 2.72 (t, J=7.5 Hz, 2H), 2.57 (t, J=7.5 Hz, 2H), 1.96 (s, 3H), 1.77-1.66 (m, 8H), 1.40-1.28 (m, 2H); ESI (m/z) [C.sub.32H.sub.49N.sub.3O.sub.8+H].sup.+ 604.

[0810] Preparation of Exp-105: To a solution of Exp-105b (173 mg, 0.239 mmol) in DMF (4.0 mL) was added compound 1 (93 mg, 0.239 mmol) and DIPEA (185 mg, 1.43 mmol) at room temperature. Solid precipitated out. The resultant mixture was concentrated at 65 C. by rotovap over 30 min. The solubility was improved during the concentration. After concentrated, DMF (4.0 mL) was added. Some sticky solid stayed at the bottom of rbf and did not go into the solution. The mixture was concentrated at 65 C. by rotovap over 30 min again. DMF (4.0 mL) was added, and a clear solution was observed. The resultant reaction mixture was stirred at 70 C. for 1 h. After the solvent was removed, the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in water, product came out at 40%), to afford Exp-105 (170 mg, 87%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.29-7.27 (m, 4H), 4.44-4.41 (m, 1H), 4.02-4.01 (m, 1H), 3.83-3.64 (m, 8H), 3.36-3.34 (m, 2H), 3.11-3.10 (m, 2H), 2.99-2.86 (m, 4H), 2.72 (t, J=7.5 Hz, 2H), 2.59 (t, J=7.5 Hz, 2H), 1.81-1.65 (m, 8H), 1.37-1.36 (m, 2H); ESI (m/z) [C.sub.38H.sub.54ClN.sub.9O.sub.9+H].sup.+ 816; HPLC, AUC=96.0%; t.sub.R=9.78 min, Method A.

Example 106

Preparation of N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-L-lysine (Exp-106)

##STR00477##

[0811] Preparation of Exp-106: To a solution of Exp-105 (107 mg, 0.131 mmol) in THF (1.0 mL), MeOH (1.0 mL) and water (1.0 mL) was added NaOH (52.4 mg, 1.31 mmol) at room temperature. The resultant mixture was stirred at room temperature for 3 h. After the solvent was removed, the residue purified by reverse phase column (5% to 90% of CH.sub.3CN in H.sub.2O, product came out at 40%), to afford compound Exp-106 (90 mg, 83%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.29-7.27 (m, 4H), 4.26-4.23 (m, 1H), 3.96-3.95 (m, 1H), 3.80-3.63 (m, 5H), 3.50-3.40 (m, 4H), 2.97-2.94 (m, 4H), 2.78-2.71 (m, 4H), 2.58 (t, J=7.5 Hz, 2H), 1.79-1.56 (m, 8H), 1.25-1.23 (m, 2H); ESI (m/z) [C.sub.37H.sub.52ClN.sub.9O.sub.9+H].sup.+ 802; HPLC, AUC=96.7%; t.sub.R=9.82 min, Method A.

Example 107

Preparation of methyl N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.,N.SUP.6.-bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-L-lysinate (Exp-107)

##STR00478##

[0812] Preparation of Exp-107a: To a solution of Int-35 (100 mg, 0.164 mmol) in MeOH (3.0 mL) was added D-glucose (148 mg, 0.819 mmol) followed by AcOH (49.2 mg, 0.819 mmol) and NaCNBH.sub.3 (51.5 mg, 0.819 mmol). The resulting solution was stirred at 50 C. for 16 h. Additional D-glucose (29.5 mg, 0.164 mmol), AcOH (9.84 mg, 0.164 mmol) and NaCNBH.sub.3 (10.5 mg, 0.164 mmol) were added, and the reaction mixture was stirred at 50 C. for 24 h. The solvent was removed, and the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in H.sub.2O, product came out at 50% to 70%). The pure fractions were collected, and solvents were removed. The residue was azeotrope with MeOH to remove residual water, then 3N HCl in MeOH (0.55 mL) was added, and azeotroped with MeOH to remove boron, to afford Exp-107a (113 mg, 73%) as a white solid: ESI (m/z) [C.sub.46H.sub.67N.sub.3O.sub.15+H].sup.+ 902.

[0813] Preparation of Exp-107b: To a solution of Exp-107a (490 mg, 0.522 mmol) in EtOH (5.0 mL) and water (5.0 mL) was added 20% Pd(OH).sub.2 (50 mg). The resultant mixture was stirred under hydrogen (balloon) at room temperature for 8 h. Around 20% conversion was observed. After filtration, fresh 20% Pd(OH).sub.2 (50 mg) was added, and the resultant suspension was stirred under hydrogen (balloon) for 16 h. Around 60% conversion was observed. The temperature was increased to 40 C., and the suspension was stirred under hydrogen for 4 h. After filtration, the solvent was removed to afford Exp-107b (429 mg, 93%) as colorless syrup: ESI (m/z) [C.sub.36H.sub.61N.sub.3O.sub.13+H].sup.+ 768.

[0814] Preparation of Exp-107: To a solution of Exp-107b (425 mg, 0.479 mmol) in DMF (6.0 mL) was added compound 1 (205 mg, 0.526 mmol) and DIPEA (370 mg, 2.87 mmol) at room temperature. The resultant mixture was warmed to 65 C. and stirred for 8 h. After solvent removed, the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in H.sub.2O, product came out at 30% to 60%), to afford Exp-107 (272 mg, 58%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.30-7.27 (m, 4H), 4.45-4.42 (m, 1H), 4.13 (br s, 2H), 3.82-3.63 (m, 13H), 3.50-3.36 (m, 8H), 2.97-2.94 (m, 2H), 2.72 (t, J=7.5 Hz, 2H), 2.59 (t, J=7.5 Hz, 2H), 1.75-1.60 (m, 8H), 1.40-1.36 (m, 2H); ESI (m/z) [C.sub.44H.sub.66ClN.sub.9O.sub.14+H].sup.+ 980; HPLC, AUC=96.6%; t.sub.R=9.00 min, Method A.

Example 108

Preparation of N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.,N.SUP.6.-bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-L-lysine (Exp-108)

##STR00479##

[0815] Preparation of Exp-108: To a solution of Exp-107 (230 mg, 0.235 mmol) in THF (3.0 mL), MeOH (3.0 mL) and water (1.0 mL) was added NaOH (94 mg, 2.35 mmol) at room temperature. The resultant mixture was stirred at room temperature for 4 h. After the solvent was removed, the residue was dissolved in water (3.0 mL), and the pH was adjusted to 7. The precipitated solid was filtered and dried, to afford 180 mg as a yellow solid. The solid was dissolved in CH.sub.3CN/water (3.0 mL/6.0 mL) at pH=2 and the solution was purified by reverse phase column (5% to 90% of CH.sub.3CN in H.sub.2O, product came out at 40%), to afford Exp-108 (134 mg, 55%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.48 (m, 4H), 7.29-7.27 (m, 4H), 4.28-4.25 (m, 1H), 4.04 (br s, 2H), 3.81-3.63 (m, 10H), 3.50-3.40 (m, 4H), 3.16-2.94 (m, 6H), 2.73 (t, J=7.5 Hz, 2H), 2.58 (t, J=7.5 Hz, 2H), 1.81-1.62 (m, 8H), 1.26-1.25 (m, 2H); ESI (m/z) [C.sub.43H.sub.64ClN.sub.9O.sub.14+H].sup.+ 966; HPLC, AUC=98.9%; t.sub.R=8.83 min, Method A.

Example 109

Preparation of methyl N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-N6-(3-phenylpropyl)-L-lysinate (Exp-109)

##STR00480##

[0816] Preparation of Exp-109a: To a solution of Int-35 (100 mg, 0.164 mmol) in MeOH (5.0 mL) was added D-glucose (32.5 mg, 0.180 mmol) followed by NaCNBH.sub.3 (15.4 mg, 0.246 mmol). The resulting solution was stirred at 50 C. for 6 h. 3-Phenylpropanal (33.0 mg, 0.246 mmol), AcOH (29.5 mg, 0.492 mmol) and NaCNBH.sub.3 (15.4 mg, 0.246 mmol) were added to the reaction mixture and the resulting solution was stirred at 50 C. for 1 h. The solvent was removed, and the resulting residue was washed with water (1.0 mL), then azeotroped with MeOH to remove water. The residue was washed with MTBE (3.0 mL), dissolved in MeOH (5.0 mL) and 3 N HCl in MeOH (1.0 mL) was added. The solution was concentrated and azeotroped with MeOH (5.0 mL3) to afford Exp-109a (117 mg, 80%) as a white solid, which was used in the next step: ESI (m/z) [C.sub.49H.sub.65N.sub.3O.sub.10+H].sup.+ 856.

[0817] Preparation of Exp-109b: To a solution of Exp-109a (117 mg, 0.131 mmol) in EtOH (2.0 mL) and water (2.0 mL) was added 20% Pd(OH).sub.2 (15 mg) and AcOH (47.2 mg, 0.787 mmol). The resultant mixture was stirred under hydrogen (balloon) at 45 C. for 6 h. After filtration, the solvent was removed, to afford Exp-109b (104 mg, 94%) as colorless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.51-7.49 (m, 4H), 7.29-7.19 (m, 9H), 4.42-4.37 (m, 1H), 3.99-3.96 (m, 1H), 3.78-3.55 (m, 8H), 3.06-2.58 (m, 16H), 1.95 (s, 6H), 1.77-1.56 (m, 10H), 1.28-1.17 (m, 2H); ESI (m/z) [C.sub.41H.sub.59N.sub.3O.sub.8+H].sup.+ 722.

[0818] Preparation of Exp-109: To a solution of Exp-109b (413 mg, 0.490 mmol) in DMF (6.0 mL) was added compound 1 (191 mg, 0.490 mmol) and DIPEA (380 mg, 2.94 mmol) at room temperature. The resultant reaction mixture was stirred at 60 C. for 6 h. After the solvent was removed, the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in water, product came out at 30% to 40%), to afford Exp-109 (255 mg, 55%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.31-7.20 (m, 9H), 4.44-4.41 (m, 1H), 4.05-4.00 (m, 1H), 3.79-3.63 (m, 8H), 3.36-3.34 (m, 2H), 3.19-2.94 (m, 8H), 2.73-2.58 (m, 6H), 2.03-1.99 (m, 2H), 1.81-1.61 (m, 8H), 1.28-1.25 (m, 2H); ESI (m/z) [C.sub.47H.sub.64ClN.sub.9O.sub.9+H].sup.+ 934; HPLC, AUC=97.2%; t.sub.R=10.16 min, Method A.

Example 110

Preparation of N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-N.SUP.6.-(3-phenylpropyl)-L-lysine (Exp-110)

##STR00481##

[0819] Preparation of Exp-110: To a solution of Exp-109 (48 mg, 0.0510 mmol) in THF (1.0 mL), MeOH (1.0 mL) and water (1.0 mL) was added NaOH (20.5 mg, 0.514 mmol) at room temperature. The resultant mixture was stirred at room temperature for 3 h. The reaction mixture was concentrated, the resulting residue was dissolved in 2 mL of water and the pH was adjusted to 5 with AcOH. Solid precipitation was observed. To the suspension was added 1 N NaOH and the resulting solution was purified using reverse phase chromatography (5%90% of CH3CN in water, product came out at 40%), to afford compound Exp-110 (52 mg) as an off-white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.28-7.20 (m, 9H), 4.25-4.23 (m, 1H), 4.05-4.00 (m, 1H), 3.50-3.40 (m, 2H), 3.78-3.62 (m, 5H), 3.10-2.94 (m, 8H), 2.73-2.57 (m, 6H), 1.95-1.93 (m, 2H), 1.80-1.57 (m, 8H), 1.25-1.20 (m, 2H); ESI (m/z) [C.sub.46H.sub.62ClN.sub.9O.sub.9+H].sup.+ 920; HPLC, AUC=96.7%; t.sub.R=10.24 min, Method A.

Example 111

Preparation of methyl N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.-hexyl-N.SUP.6.-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-L-lysinate (Exp-111)

##STR00482##

[0820] Preparation of Exp-111: In a three-step procedure Exp-111 was synthesized beginning with Int-35 using a sequence analogous to the one used to prepare Exp-109 substituting n-hexanal for 3-phenylpropanal: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.49 (m, 4H), 7.30-7.27 (m, 4H), 4.27-4.24 (m, 1H), 3.91 (br s, 1H), 3.80-3.63 (m, 8H), 3.50-3.40 (m, 3H), 2.98-2.58 (m, 11H), 1.82-1.10 (m, 18H), 1.00-0.90 (m, 3H); ESI (m/z) [C.sub.44H.sub.66ClN.sub.9O.sub.9+H].sup.+ 900; HPLC, AUC=94.7%; t.sub.R=10.51 min, Method A.

Example 112

Preparation of N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.-hexyl-N.SUP.6.-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-L-lysinate (Exp-112)

##STR00483##

[0821] Preparation of Exp-112: In a one-step procedure Exp-112 was synthesized beginning with Exp-111 using an analogous procedure to the one used to prepare Exp-110: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.54-7.50 (m, 4H), 7.30-7.28 (m, 4H), 4.45-4.42 (m, 1H), 4.10-4.08 (m, 1H), 3.81-3.64 (m, 8H), 3.76-3.08 (m, 7H), 2.96 (t, J=7.5 Hz, 2H), 2.73 (t, J=7.5 Hz, 2H), 2.02-1.64 (m, 10H), 1.35-1.32 (m, 8H), 1.00-0.90 (m, 3H); ESI (m/z) [C.sub.43H.sub.64ClN.sub.9O.sub.9+H].sup.+ 886; HPLC, AUC=99.8%; t.sub.R=10.31 min, Method A.

Examples 113-116

[0822] Preparation of Exp-113-116: In an analogous sequence of procedures used for Examples 105-108, Examples 113-116 were synthesized beginning with Int-35 and substituting D-lysine for L-lysine:

##STR00484##

TABLE-US-00038 Exp # R.sup.9 R R.sup.1 Spectra 113 CH.sub.3 Sug H .sup.1H NMR (500 MHz, CD.sub.3OD) 7.54-7.50 (m, 4H), 7.30-7.27 (m, 4H), 4.43-4.39 (m, 1H), 4.02-4.00 (m, 1H), 3.83-3.62 (m, 8H), 3.50-3.40 (m, 2H), 3.33-3.32 (m, 2H), 3.10-3.08 (m, 2H), 2.97- 2.90 (m, 4H), 2.75-2.71 (m, 2H), 1.93-1.65 (m, 8H), 1.37-1.32 (m, 2H); ESI (m/z) [C.sub.38H.sub.54ClN.sub.9O.sub.9 + H].sup.+ 816; HPLC, AUC = 95.0%; t.sub.R = 8.86 min, Method A. 114 H Sug H .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.28-7.26 (m, 4H), 4.25-4.23 (m, 1H), 3.98-3.96 (m, 1H), 3.80-3.62 (m, 5H), 3.50-3.40 (m, 2H), 3.16-2.78 (m, 6H), 2.72 (t, J = 7.5 Hz, 2H), 2.58 (t, J = 7.5 Hz, 2H), 1.79-1.60 (m, 8H), 1.30-1.20 (m, 2H); ESI (m/z) [C.sub.37H.sub.52ClN.sub.9O.sub.9 + H].sup.+ 802; HPLC, AUC = 95.5%; t.sub.R = 8.56 min, Method A. 115 CH.sub.3 Sug Sug .sup.1H NMR (500 MHz, D.sub.2O) 7.53-7.50 (m, 4H), 7.30-7.27 (m, 4H), 4.46-4.43 (m, 1H), 4.08 (br s, 2H), 3.81-3.63 (m, 12H), 3.50-3.40 (m, 7H), 3.37-3.30 (m, 2H), 2.97-2.94 (m, 2H), 2.73 (t, J = 7.0 Hz, 2H), 2.59 (t, J = 7.0 Hz, 2H), 1.85-1.66 (m, 8H), 1.38-1.28 (m, 2H); ESI (m/z) [C.sub.44H.sub.66ClN.sub.9O.sub.14 + H].sup.+ 980; HPLC, AUC = 97.9%; t.sub.R = 8.64 min, Method A. 116 H Sug Sug .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.47 (m, 4H), 7.28-7.26 (m, 4H), 4.26-4.23 (m, 1H), 3.84-3.61 (m, 11H), 3.50-3.40 (m, 3H), 2.95 (t, J = 7.5 Hz, 2H), 2.72 (t, J = 7.0 Hz, 2H), 2.59-2.47 (m, 8H), 1.80-1.37 (m, 8H), 1.20-1.10 (m, 2H); ESI (m/z) [C.sub.43H.sub.64ClN.sub.9O.sub.14 + H].sup.+ 966; HPLC, AUC = 96.5%; t.sub.R = 8.42 min, Method A.

Example 117

Preparation of methyl (S)-3,5-diamino-6-chloro-N(N-(4-(4-(3-((1,6-diamino-1-oxohexan-2-yl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-117)

##STR00485##

[0823] Preparation of Exp-117a: To a stirred solution of Int-50a (850 mg, 1.29 mmol) in IPA:EtOAc:H.sub.2O (30 mL:30 mL:30 mL) and CH.sub.3CO.sub.2H (0.8 mL) was charged 20% Pd(OH).sub.2 on carbon 50% wet (320 mg, 50% weight substrate) at room temperature. The resultant reaction mixture was stirred under H.sub.2 (balloon) at room temperature for 16 h. The reaction mixture was filtered through celite bed and filtrate concentrated under reduced pressure. The obtained crude was azeotroped with toluene (20 mL) to afford Exp-117a (600 mg, 79%) as an off-white solid: ESI (m/z) [C.sub.30H.sub.44N.sub.4O.sub.4+H].sup.+ 525.

[0824] Preparation of Exp-117b: A stirred solution of Exp-117a (600 mg, 1.027 mmol) and compound 1 (390 mg, 1.027 mmol) in DMF (10 mL) was charged with DIPEA (0.67 mL, 4.10 mmol) at room temperature. The resultant reaction mixture was heated to 60 C. and stirred for 16 h. The reaction mixture was poured into a beaker containing cold water (50 mL), the resulting solids were collected by filtration then washed with water (10 mL) followed by hexanes (20 mL). The obtained solid was dried under reduced pressure to afford Exp-117b (630 mg, 83%) as a yellow solid: ESI (m/z) [C.sub.36H.sub.49ClN.sub.10O.sub.5+H].sup.+ 737.

[0825] Preparation of Exp-117: A stirred solution of Exp-117b (610 mg, 0.827 mmol) in 1,4-dioxane (5.0 mL), was charged with 4M HCl in 1,4-dioxane (5.0 mL) followed by water (1.0 mL) then stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure. The obtained crude compound was purified by reverse phase comb-flash chromatography eluting with (10%-100% CH.sub.3CN in water) to afford Exp-117 (300 mg, 78%) as a light brown solid: .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.42 (dd, J=5.2, 8.0 Hz, 4H), 7.18 (d, J=8.0 Hz, 4H), 4.31 (dd, J=5.2, 8.8 Hz, 1H), 3.26 (t, J=6.8 Hz, 2H), 2.88-2.84 (m, 2H), 2.79-2.74 (m, 2H), 2.63 (t, J=6.8 Hz, 2H), 2.51 (t, J=7.6 Hz, 2H), 1.71-1.64 (m, 5H), 1.55-1.1.50 (m, 3H), 1.43-1.25 (m, 2H); ESI (m/z) [C.sub.31H.sub.41ClN.sub.10O.sub.3+H].sup.+ 637; HPLC, AUC=99.7%; t.sub.R=6.33 min, Method I.

Example 118

Preparation of (R)-3,5-diamino-6-chloro-N(N-(4-(4-(3-((1,6-diamino-1-oxohexan-2-yl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-118)

##STR00486##

[0826] Preparation of Exp-118: The R-enantiomer of Exp-35a was converted to Exp-118 in a sequence of reactions analogous to those used to prepare Exp-118: .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.43-7.40 (m, 4H), 7.19 (d, J=8.0 Hz, 4H), 4.24-4.20 (m, 1H), 3.26 (t, J=6.4 Hz, 2H), 2.88-2.84 (m, 2H), 2.77 (t, J=6.4 Hz, 2H), 2.63 (t, J=6.8 Hz, 2H), 2.51 (t, J=7.6 Hz, 2H), 1.71-1.60 (m, 5H), 1.58-1.50 (m, 3H), 1.40-1.32 (m, 2H); ESI (m/z) [C.sub.31H.sub.41ClN.sub.10O.sub.3+H].sup.+ 637; HPLC, AUC=99.9%; t.sub.R=6.32 min, Method I.

Example 119

Preparation of (S)-3,5-diamino-6-chloro-N(N-(4-(4-(3-((1,6-diamino-1-oxohexan-2-yl) (methyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-119)

##STR00487##

[0827] Preparation of Exp-119: Example-119 was synthesized starting with Int-23 and Int-46 and used a sequence of step analogous to the preparation of Example 117. .sup.1H NMR (500 MHz, CD.sub.3OD) 7.51-7.50 (m, 4H), 7.31-7.26 (m, 4H), 5.09-5.06 (m, 1H), 3.16-2.66 (m, 2H), 3.16-2.66 (m, 11H), 1.92-1.91 (m, 1H), 1.79-1.49 (m, 7H), 1.25-1.20 (m, 2H); ESI (m/z) [C.sub.32H.sub.43ClN.sub.10O.sub.3+H].sup.+ 651; HPLC, AUC=97.7%; t.sub.R=9.48 min, Method A.

Example 120

Preparation of methyl N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.2.-methyl-N.SUP.6.-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-L-lysinate (Exp-120)

##STR00488##

[0828] Preparation of Exp-120a: To a solution of Exp-20a (2.30 g, 3.34 mmol) in THF (20 mL) was added 4N HCl in dioxane (20 mL). The reaction mixture was stirred at rt for 2 h. After the solvent was removed, the residue was crashed from MTBE/hexanes (20 mL/5 mL). The solvent was decanted and the residue was dried over high vacuum, to afford Exp-120a (1.94 g, 93%) as a yellow solid: ESI (m/z) [C.sub.35H.sub.45N.sub.3O.sub.5+H].sup.+ 588.

[0829] Preparation of Exp-120b: To a solution of Exp-120a (100 mg, 0.160 mmol) in MeOH (3.0 mL) was added D-glucose (31.7 mg, 0.176 mmol), NaCNBH.sub.3 (11.0 mg, 0.176 mmol) and AcOH (10.5 mg, 0.117 mmol). The resulting solution was stirred at 45 C. for 16 h. HPLC showed 80% of desired mono was formed along with 8% of bis and 12% of starting material Exp-120a. After the solvent was removed, the residue was purified by reverse phase column (10 to 90% of CH.sub.3CN in water followed by 90% of MeOH in water, product dragging from 60% of CH.sub.3CN in water till end). The pure fractions were collected, concentrated, azeotroped with MeOH (3 mL) to remove residual water. The residue was suspended in MeOH (3.0 mL), 3 N HCl in MeOH (0.15 mL) was added and the solution was concentrated to remove boron. The residue was azeotroped with MeOH (3.0 mL) and 3 HCl in MeOH (0.15 mL) again, to afford Exp-120b (64 mg, 90%) as a colorless syrup: ESI (m/z) [C.sub.41H.sub.57N.sub.3O.sub.10+H].sup.+ 752.

[0830] Preparation of Exp-120c: To a solution of Exp-120b (450 mg, 0.571 mmol) in MeOH (10 mL) was added Pd(OH).sub.2/C (100 mg) and the resulting suspension was stirred under hydrogen (balloon) at rt for 3 h, then 40 C. for 2 h. After filtration, the solvent was removed and the residue was dried over high vacuum to afford Exp-120c (354 mg, 95%) as a colorless syrup: ESI (m/z) [C.sub.33H.sub.51N.sub.3O.sub.8+H].sup.+ 618.

[0831] Preparation of Exp-120: To a solution of Exp-120c (352 mg, 0.538 mmol) in DMF (8.0 mL) was added compound 1 (209 mg, 0.538 mmol) followed by DIPEA (278 mg, 2.15 mmol). The resultant solution was stirred at 55 C. for 16 h. After the solvent was removed, the residue was purified by reverse phase column (10% to 60% of CH.sub.3CN in water, product came out at 40%), to afford Exp-120 (228 mg, 51%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.31-7.28 (m, 4H), 5.04-5.02 (m, 1H), 4.04-4.02 (m, 1H), 3.84-3.63 (m, 8H), 3.37-3.32 (m, 3H), 3.14-3.13 (m, 2H), 2.99-2.95 (m, 6H), 2.80-2.72 (m, 4H), 2.03-2.00 (m, 1H), 1.81-1.70 (m, 7H), 1.40-1.32 (m, 2H); ESI (m/z) [C.sub.39H.sub.56ClN.sub.9O.sub.9+H].sup.+ 830; HPLC, AUC=97.4%; t.sub.R=9.70 min, Method A.

Example 121

Preparation of N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.2.-methyl-N.SUP.6.-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-L-lysine (Exp-121)

##STR00489##

[0832] Preparation of Exp-121: To a solution of Exp-120 (118 mg, 0.142 mmol) in THF/MeOH/H2O (0.6 mL/0.6 mL/0.2 mL) was added NaOH (28.4 mg, 0.710 mmol). The resultant solution was stirred at rt for 16 h. After pH was adjusted to 2 with 1 N HCl, the solvent was removed and the residue was purified by reverse phase column (10% to 80% of CH.sub.3CN in water, product came out at 20 to 40%), to afford Exp-121 (88 mg, 69%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.54-7.51 (m, 4H), 7.33-7.29 (m, 4H), 5.10-5.07 (m, 1H), 4.06-4.04 (m, 1H), 3.83-3.65 (m, 5H), 3.37-3.35 (m, 3H), 3.17-3.12 (m, 2H), 3.00-2.97 (m, 6H), 2.83-2.72 (m, 4H), 2.03-2.01 (m, 1H), 1.84-1.74 (m, 7H), 1.40-1.33 (m, 2H); ESI (m/z) [C.sub.38H.sub.54ClN.sub.9O.sub.9+H].sup.+ 816; HPLC, AUC=96.5%; t.sub.R=9.56 min, Method A.

Example 122

Preparation of methyl N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N2-methyl-N.SUP.6.,N.SUP.6.-bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-L-lysinate (Exp-122)

##STR00490##

[0833] Preparation of Exp-122a: To a solution of Exp-120a (100 mg, 0.160 mmol) in MeOH (3.0 mL) was added D-glucose (87.0 mg, 0.481) followed by NaCNBH.sub.3 (30.2 mg, 0.481 mmol) and AcOH (28.9 mg, 0.481 mmol). The resultant solution was stirred at 60 C. for 16 h. Additional D-glucose (29.0 mg, 0.160 mmol), NaCNBH.sub.3 (10.0 mg, 0.160 mmol) and AcOH (1.00 mg, 0.160 mmol) were added. The resultant solution was stirred at 60 C. for 6 h. After the solvent was removed, water (3.0 mL) and MeOH (1.5 mL) were added. The resulting solid was collected by filtration, which became a sticky syrup after filtration. The syrup was azeotroped with MeOH (10 mL) to remove water, then re-suspended in MeOH (10 mL) and 3 N HCl in MeOH (0.2 mL) was added resulting in a clear solution. The solution was concentrated, and the above procedure was repeated 2 times, to afford 112 mg of material. The filtrate from earlier step was purified by reverse phase column (10% to 90% of MeOH in water, product came out at 80%), to afford additional 14 mg of material. A total 126 mg of compound Exp-122a was obtained as colorless syrup with 83% yield: ESI (m/z) [C.sub.47H.sub.69N.sub.3O.sub.15+H].sup.+ 916.

[0834] Preparation of Exp-122b: To a solution of Exp-122a (985 mg, 1.034 mmol) in MeOH (20 mL) was added Pd(OH).sub.2/C (150 mg). The resulting suspension was stirred under hydrogen (balloon) at rt for 3 h, warmed to 40 C. and stirred an additional 5 h. After filtration, solvent was removed and the residue was dried over high vacuum, to afford Exp-122b (832 mg, 98%) as a colorless syrup: ESI (m/z) [C.sub.39H.sub.63N.sub.3O.sub.13+H]782.

[0835] Preparation of Exp-122: To a solution of Exp-122b (830 mg, 1.10 mmol) in DMF (12 mL) was added 1 (394 mg, 1.014 mmol) followed by DIPEA (524 mg, 4.06 mmol). The resultant solution was stirred at 55 C. for 16 h. After the solvent was removed, the residue was purified by reverse phase column (10% to 60% of CH.sub.3CN in water, product came out at 40%), to afford Exp-122 (826 mg, 82%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.51 (m, 4H), 7.32-7.28 (m, 4H), 5.07-5.04 (m, 1H), 4.17-4.13 (m, 2H), 3.82-3.63 (m, 13H), 3.50-3.40 (m, 8H), 2.98-2.95 (m, 5H), 2.80-2.72 (m, 4H), 2.03-2.00 (m, 1H), 1.87-1.72 (m, 7H), 1.40-1.35 (m, 2H); ESI (m/z) [C.sub.45H.sub.68ClN.sub.9O.sub.14+H].sup.+ 994; HPLC, AUC=98.4%; t.sub.R=9.55 min, Method A.

Example 123

Preparation of N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N2-methyl-N.SUP.6.,N.SUP.6.-bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-L-lysine (Exp-123)

##STR00491##

[0836] Preparation of Exp-123: To a solution of Exp-122 (160 mg, 0.161) in THF/MeOH/H.sub.2O (1.5 mL/1.5 mL/0.5 mL) was added NaOH (32.2 mg, 0.804 mmol). The resultant solution was stirred at rt for 3 h. After neutralization with 1 N HCl, the solvent was removed. Water (2.0 mL) was added, the resulting solid was collected by filtration (filtrate retained) and washed with water (1.0 mL). The solid was dissolved in CH.sub.3CN/water (1 mL/2 mL) and added back to the retained filtrate. The pH of the solution was adjusted to 2 with 1 N HCl, the solution was purified using reverse phase column chromatography (10% to 60% of CH.sub.3CN in water, product came out from 20% to 40%), to afford Exp-123 (123 mg, 78%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.54-7.50 (m, 4H), 7.33-7.28 (m, 4H), 5.09-5.07 (m, 1H), 4.18-4.15 (m, 2H), 3.83-3.64 (m, 10H), 3.50-3.40 (m, 5H), 3.36-3.34 (m, 4H), 2.98-2.72 (m, 8H), 2.04-2.02 (m, 1H), 1.81-1.74 (m, 7H), 1.40-1.31 (m, 2H); ESI (m/z) [C.sub.44H.sub.66ClN.sub.9O.sub.14+H].sup.+ 980; HPLC, AUC=98.7%; t.sub.R=9.43 min, Method A.

Example 124

Preparation of 3,5-diamino-N(N-(4-(4-(3-(((S)-1-amino-1-oxo-6-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)hexan-2-yl)(methyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-124)

##STR00492##

[0837] Preparation of Exp-124a: Exp-120b (328 mg, 0.436 mmol) was dissolved in 7 N NH3 in MeOH (10 mL), warmed 40 C. and stirred and stirred for 2 days. The solvent was removed to afford Exp-124a (318 mg, crude) as a yellow syrup: ESI (m/z) [C.sub.40H.sub.56N.sub.4O.sub.9+H].sup.+ 737.

[0838] Preparation of Exp-124b: To a solution of Exp-124a (367 mg) in MeOH (8.0 mL) was added Pd(OH).sub.2/C (80 mg). The resultant solution was stirred at 45 C. under hydrogen (balloon) for 4 h. After filtration, the solvent was removed, to afford Exp-124b (275 mg, crude) as a colorless syrup: ESI (m/z) [C.sub.32H.sub.50N.sub.4O.sub.7+H].sup.+ 603.

[0839] Preparation of Exp-124: To a solution of compound Exp-124b (275 mg, 0.456 mmol) in DMF (8.0 mL) was added 1 (177 mg, 0.456 mmol) followed by DIPEA (236 mg, 1.82 mmol). The resultant solution was stirred at 55 C. for 16 h. After the solvent was removed, the residue was washed with EtOH, then purified by reverse phase column (10% to 80% of CH.sub.3CN in water, product came out at 30% to 50%), to afford Exp-124 (98 mg, 25% over 3 steps) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.51 (m, 4H), 7.32-7.28 (m, 4H), 5.12-5.09 (m, 1H), 4.02-4.01 (m, 1H), 3.83-3.63 (m, 5H), 3.35-3.32 (m, 2H), 3.16-3.10 (m, 2H), 2.98-2.72 (m, 11H), 2.02-1.95 (m, 1H), 1.79-1.68 (m, 7H), 1.32-1.27 (m, 2H); ESI (m/z) [C.sub.38H.sub.55ClN.sub.10O.sub.5+H].sup.+ 815; HPLC, AUC=94.3%; t.sub.R=9.29 min, Method A.

Example 125

Preparation of 3,5-diamino-N(N-(4-(4-(3-(((S)-1-amino-6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)-1-oxohexan-2-yl)(methyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-125)

##STR00493##

[0840] Preparation of Exp-125: Exp-120a was converted to Exp-125 in an analogous sequence of steps used in Example-124: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.51 (m, 4H), 7.32-7.28 (m, 4H), 5.13-5.10 (m, 1H), 4.09 (br s, 2H), 3.81-3.63 (m, 10H), 3.50-3.20 (m, 8H), 3.00-2.72 (m, 9H), 1.96-1.95 (m, 1H), 1.78-1.73 (m, 7H), 1.28-1.27 (m, 2H); ESI (m/z) [C.sub.44H.sub.67ClN.sub.10O.sub.13+H].sup.+ 979; HPLC, AUC=98.9%; t.sub.R=9.01 min, Method A.

Example 126

Preparation of methyl N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.-(2-hydroxyethyl)-N2-methyl-L-lysinate (Exp-126)

##STR00494##

[0841] Preparation of Exp-126a: To a solution of Exp-120a (50 mg, 0.080 mmol) and benzaldehyde (8.50 mg, 0.080 mmol) in MeOH (2.0 mL) was added NaCNBH.sub.3 (5.00 mg, 0.080 mmol) and AcOH (4.98 mg, 0.080 mmol) at rt. The reaction mixture was stirred at 45 C. for 3 h. Additional benzaldehyde (4.25 mg, 0.040 mmol), NaCNBH.sub.3 (4.25 mg, 0.040 mmol) and AcOH (2.49 mg, 0.040 mmol) were added. The reaction mixture was stirred at 45 C. for 2 h. Additional benzaldehyde (4.25 mg, 0.040 mmol), NaCNBH.sub.3 (4.25 mg, 0.040 mmol) and AcOH (2.49 mg, 0.040 mmol) were added. The reaction mixture was stirred at 45 C. for 2 h. LCMS suggest 80% of mono-benzyl amide derivative was formed along with 10% of bis and 10% of starting material Exp-120a. 1,4-Dioxane-2,5-diol (3.0 equiv), NaCNBH.sub.3 (6.0 equiv), and AcOH (6.0 equiv) were added. The reaction mixture was stirred at 55 C. for 16 h. LCMS suggested 80% of desired product Exp-126a was formed. After the solvent was removed, the residue was purified by reverse phase column (10% to 90% of CH.sub.3CN in water, product came out at 40% to 60%). The pure fractions were combined, concentrated, azeotroped with 1 N HCl in MeOH (3.0 mL2) and MeOH (3.0 mL), to afford Exp-126a (36.5 mg, 60%) as a yellow syrup: ESI (m/z) [C.sub.44H.sub.55N.sub.3O.sub.6+H].sup.+ 722.

[0842] Preparation of Exp-126b: To a solution of Exp-126a (320 mg, 0.422 mmol) in MeOH (10 mL) was added Pd(OH).sub.2 (60 mg) at rt. The reaction mixture was stirred under hydrogen (balloon) at 45 C. for 3 h. After filtration, the solvent was removed, to afford Exp-126b (210 mg, 93%) as a yellow syrup: ESI (m/z) [C.sub.38H.sub.62N.sub.4O.sub.12+H].sup.+ 766.

[0843] Preparation of Exp-126: To a solution of compound Exp-126b (170 mg, 0.318 mmol) in DMF (5.0 mL) was added compound 1 (124 mg, 0.318 mmol) and DIPEA (123 mg, 0.955 mmol). The resulting solution was stirred at 55 C. for 16 hr. After the solvent was removed, the residue was crashed from IPA (3.0 mL). After solvent decanted, the residue was purified by reverse phase column (10% to 90% CH3CN in water, product came out at 30%), to afford Exp-126 (198 mg, 88%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.51 (m, 4H), 7.31-7.28 (m, 4H), 5.05-5.02 (m, 1H), 3.76-3.74 (m, 2H), 3.70 (s, 3H), 3.44-3.32 (m, 3H), 3.05-2.93 (m, 8H), 2.80-2.71 (m, 4H), 2.01-1.99 (m, 1H), 1.83-1.66 (m, 7H), 1.34-1.29 (m, 2H); ESI (m/z) [C.sub.35H.sub.48ClN.sub.9O.sub.5+H].sup.+ 710; HPLC, AUC=94.9%; t.sub.R=10.36 min, Method A.

Example 127

Preparation of N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.6.-(2-hydroxyethyl)-N2-methyl-L-lysine (Exp-127)

##STR00495##

[0844] Preparation of Exp-127: To a solution of Exp-126 (120 mg, 0.169 mmol) in MeOH (2.0 mL) and water (2.0 mL) was added NaOH (67.6 mg, 1.68 mmol). The yellow solution was stirred at rt for 1 h. The pH was adjusted to 2, then the solvent was removed and the residue was purified by reverse phase column (5% to 80% of CH.sub.3CN in water, product came out at 20%), to afford Exp-127 (98 mg, 75%) as a yellow solid: 7.52-7.47 (m, 4H), 7.31-7.27 (m, 4H), 5.05-5.01 (m, 0.71H), 4.21-4.18 (m, 0.3H), 3.76-3.72 (m, 2H), 3.40-3.30 (m, 3H), 3.03-2.88 (m, 8H), 2.77-2.71 (m, 4H), 2.02-1.98 (m, 1H), 1.81-1.64 (m, 7H), 1.28-1.24 (m, 2H); ESI (m/z) [C.sub.34H.sub.46ClN.sub.9O.sub.5+H].sup.+ 696; HPLC, AUC=96.2%; t.sub.R=10.01 min, Method A.

Examples 128-134

Preparation of Exp-128-134: In an analogous sequence of procedures used for preparation of Examples 20-21, and Examples 119-123, D-N(CH.SUB.3.)-lysine was substituted into the sequence for the preparation of Examples 128-134

##STR00496##

TABLE-US-00039 Exp # R.sup.2 R R.sup.1 Spectra 128 CO.sub.2CH.sub.3 H H .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.43-7.40 (m, 4H), 7.22- 7.178 (m, 4H), 4.97-4.88 (m, 1H), 3.61 (d, J = 8.08 Hz, 3H), 3.28-3.22 (m, 3H), 2.87 (s, 3H), 2.86-2.83 (m, 1H), 2.83-2.76 (m, 2H), 2.71-2.67 (m, 2H), 2.66-2.60 (m, 2H), 1.96-1.84 (m, 1H), 1.78-1.52 (m, 7H), 1.31-1.12 (m,2H); ESI (m/z) [C.sub.33H.sub.44ClN.sub.9O.sub.4 + H].sup.+ 666; HPLC, AUC = 99.4%; t.sub.R = 8.50 min, Method W. 129 CO.sub.2H H H .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.43-7.40 (m, 4H), 7.22- 7.178 (m, 4H), 5.03-4.97 (m, 1H), 3.26 (t, J = 6.38, 2H), 2.88 (s, 3H), 2.86-2.77 (m, 3H), 2.73-2.61 (m, 5H), 1.98-1.88 (m, 1H), 1.77-1.52 (m, 7H), 1.30-1.18 (m, 2H); ESI (m/z) [C.sub.32H.sub.42ClN.sub.9O.sub.4 + H].sup.+ 652; HPLC, AUC = 98.5%; t.sub.R = 6.40 min, Method I. 130 CONH.sub.2 H H .sup.1H NMR (400 MHz, CD.sub.3OD) 7.43-7.41 (m, 4H), 7.22-7.18 (m, 4H), 5.03-5.01 (m, 1H), 3.28-3.24 (m, 2H), 2.87-2.75 (m, 7H), 2.73-2.62 (m, 4H), 1.90-1.83 (m, 1H), 1.66-1.55 (m, 7H), 1.26-1.1.22 (m, 2H); ESI (m/z) [C.sub.32H.sub.43ClN.sub.10O.sub.3 + H].sup.+ 651; HPLC, AUC = 98.7%; t.sub.R = 7.03 min, Method Q. 131 CO.sub.2CH.sub.3 Sug H .sup.1H NMR (400 MHz, CD.sub.3OD) 7.47-7.39 (m, 4H), 7.25-7.18 (m, 4H), 5.03-4.90 (m, 1H), 4.04-3.96 (m, 1H), 3.80-3.69 (m, 2H), 3.68-3.56 (m, 5H), 3.33-3.25 (m, 2H), 3.12-3.01 (m, 2H), 2.97-2.82 (m, 7H), 2.73-2.61 (m, 5H), 2.02-1.86 (m, 1H), 1.82- 1.53 (m, 7H), 1.34-1.14 (m, 2H); ESI (m/z) [C.sub.39H.sub.56ClN.sub.9O.sub.9 + H].sup.+ 830; HPLC, AUC = 96.1%; t.sub.R = 7.07 min, Method R. 132 CO.sub.2H Sug H .sup.1H NMR (400 MHz, DMSO-d.sub.6, D.sub.2O exchange ) 7.59-7.48 (m, 4H), 7.37-7.23 (m, 4H), 4.88-4.79 (m, 0.6H), 4.14-4.03 (m, 0.3H), 3.93-3.86 (m, 1H), 3.63-3.56 (m, 2H), 3.53-3.38 (m, 3H), 3.31-3.22 (m, 2H), 3.17-3.01 (m, 2H), 2.97-2.79 (m, 6H), 2.71-2.60 (m, 5H), 1.92-1.9 (m, 1H), 1.71-1.50 (m, 7H), 1.34- 1.14 (m, 2H); ESI (m/z) [C.sub.38H.sub.54ClN.sub.9O.sub.9 + H].sup.+ 816; HPLC, AUC = 96.6%; t.sub.R = 6.97 min, Method R. 133 CO.sub.2CH.sub.3 Sug Sug .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.46-7.38 (m, 4H), 7.26-7.16 (m, 4H), 5.04-4.92 (m, 1H), 4.12-4.01 (m, 2H), 3.77-3.65 (m, 4H), 3.64-3.52 (m, 8H), 3.40-3.31 (m, 2H), 3.31-3.24 (m, 6H), 3.20-3.17 (m, 1H), 2.91-2.87 (m, 5H), 2.72 (t, J = 11 Hz, 2H), 2.63 (t, J = 6.4 Hz, 2H), 1.98-1.83 (m, 1H), 1.80-1.57 (m, 7H), 1.37-1.18 (m, 2H); ESI (m/z) [C.sub.45H.sub.68ClN.sub.9O.sub.14 + H].sup.+ 994; HPLC, AUC = 99.1%; t.sub.R = 9.99 min, Method Y. 134 CO.sub.2H Sug Sug .sup.1H NMR (400 MHz, D.sub.2O) 7.28-7.19 (m, 4H), 7.05-6.83 (m, 4H), 4.80-4.78 (m, 1H), 4.10-3.99 (m, 2H), 3.75-3.65 (m, 6H), 3.60-3.49 (m, 4H), 3.30-2.85 (m, 8H), 2.75-2.18 (m, 5H), 2.60- 2.40 (m, 4H), 1.86-1.72 (m, 1H), 1.60-1.38 (m, 7H), 0.98-0.80 (m,2H); ESI (m/z) [C.sub.44H.sub.66ClN.sub.9O.sub.14 + H].sup.+ 980; HPLC, AUC = 99.3%; t.sub.R = 9.95 min, Method Y.

Example 135

Preparation of 3,5-diamino-N(N-(4-(4-(3-((2-aminoethyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-135)

##STR00497##

[0845] Preparation of Exp-135a: To a solution of Int-34a (312 mg, 0.544 mmol) in IPA (15 mL) and water (1.0 mL) was added 20% Pd(OH).sub.2 (50 mg) and AcOH (32.7 mg, 0.544 mmol). The resultant mixture was stirred under hydrogen (balloon) at 45 C. for 4 h. After filtration, the solvent was removed, to afford Exp-135a (256 mg, 94%) as colorless syrup: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.28-7.25 (m, 4H), 3.21-3.19 (m, 2H), 3.07 (t, J=6.0 Hz, 2H), 2.95-2.90 (m, 4H), 2.71 (t, J=7.5 Hz, 2H), 2.49 (t, J=7.5 Hz, 2H), 1.89 (s, 3H), 1.75-1.67 (m, 4H), 1.40 (s, 9H); ESI (m/z) [C.sub.26H.sub.37N.sub.3O.sub.3+H].sup.+ 440.

[0846] Preparation of Exp-135b: To a solution of Exp-135a (256 mg, 0.512 mmol) in DMF (3.0 mL) and EtOH (3.0 mL) was added compound 1 (199 mg, 0.512 mmol) at room temperature. The resultant mixture was stirred at 65 C. for 6 h. After the solvent was removed, the residue was dissolved in EtOH (5.0 mL) then water (20 mL) was added. The resulting solid was filtered and dried to afford Exp-135b (308 mg, 92%) as a yellow solid: ESI (m/z) [C.sub.32H.sub.42ClN.sub.9O.sub.4+H].sup.+ 652.

[0847] Preparation of Exp-135: To a solution of Exp-135b (235 mg, 0.360 mmol) in MeOH (5.0 mL) was added 4 N HCl in dioxane (5.0 mL) at room temperature. The resultant mixture was stirred at room temperature for 3 h. After the solvent was removed to the residue was added water (2.0 mL) and CH.sub.3CN (8.0 mL). The solid was filtered, washed with CH.sub.3CN and dried to afford compound Exp-135 (195 mg, 87%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.50 (m, 4H), 7.29-7.26 (m, 4H), 3.43-3.32 (m, 4H), 3.02-2.94 (m, 4H), 2.73 (t, J=7.0 Hz, 2H), 2.56 (t, J=7.5 Hz, 2H), 1.79-1.74 (m, 4H); ESI (m/z) [C.sub.27H.sub.34ClN.sub.9O.sub.2+H].sup.+ 552; HPLC, AUC=97.2%; t.sub.R=8.66 min, Method A.

Example 136

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(3-oxo-3-((2-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)amino)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-136)

##STR00498##

[0848] Preparation of Exp-136a: To a solution of Int-34 (50 mg, 0.085 mmol) in MeOH (2.0 mL) was added D-glucose (16.1 mg, 0.089 mmol) followed by AcOH (6.13 mg, 0.102 mmol) and NaCNBH.sub.3 (6.42 mg, 0.102 mmol). The resulting solution was stirred at 50 C. for 6 h. The solvent was removed and the residue was washed satd. Na.sub.2CO.sub.3 (1.0 mL), water (1.0 mL) then azeotroped with MeOH (3 mL) to remove water. To the residue was added 3N HCl in MeOH (0.5 mL) followed by azeotroping with MeOH (3.0 mL2) to remove boron, to afford Exp-136a (51 mg, 89%) as a white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.46 (m, 4H), 7.31-7.21 (m, 9H), 5.04 (s, 2H), 4.20 (br s, 1H), 4.10 (br s, 1H), 3.85-3.42 (m, 8H), 3.20-2.93 (m, 6H), 2.65-2.54 (m, 4H), 1.66-1.51 (m, 4H); ESI (m/z) [C.sub.35H.sub.47N.sub.3O.sub.8+H].sup.+ 638.

[0849] Preparation of 136b: To a solution of Exp-136a (50 mg, 0.074 mmol) in EtOH (3.0 mL) and water (1.0 mL) was added 20% Pd(OH).sub.2 (10 mg) and AcOH (26.7 mg, 0.445 mmol). The resultant mixture was stirred under hydrogen (balloon) at 50 C. for 6 h. After filtration, the solvent was removed, to afford Exp-136b (45 mg, 97%) as colorless syrup: ESI (m/z) [C.sub.27H.sub.41N.sub.3O.sub.6+H].sup.+ 504.

[0850] Preparation of Exp-136: To a solution of Exp-136b (215 mg, 0.345 mmol) in DMF (4.0 mL) and water (2.0 mL) was added compound 1 (268 mg, 0.689 mmol) at room temperature. The resultant mixture was stirred at 60 C. for 3 h. Additional compound 1 (134 mg, 0.345 mmol) was added, and the resultant solution was stirred at 60 C. for 5 h. Additional compound 7 (134 mg, 0.345 mmol) was added, and the resultant solution was stirred at 60 C. for 16 h. After solvent was removed, 1N HCl (10 mL) was added, some undissolved solid was filtered and washed with CH.sub.3CN/water (1:1, 20 mL). The combined filtrate was concentrated and purified by reverse phase column (5% to 80% of CH.sub.3CN in water, product came out at 30% to 40%). The combined fractions were concentrated to around 15 mL then solid precipitation was observed. Saturated NaHCO.sub.3 (6.0 mL) was added to the suspension and the solid was filtered out. The solid was re-dissolved in 1N HCl (3.0 mL) and purified by reverse phase column (5% to 80% of CH3CN in water, product came out at 30% to 40%), to afford Exp-136 (69 mg, 25%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.51 (m, 4H), 7.29-7.27 (m, 4H), 4.05-4.04 (m, 1H), 3.87-3.65 (m, 5H), 3.47-3.44 (m, 4H), 3.22-3.11 (m, 4H), 2.96 (t, J=7.5 Hz, 2H), 2.73 (t, J=7.5 Hz, 2H), 2.56 (t, J=7.5 Hz, 2H), 1.79-1.74 (m, 4H); ESI (m/z) [C.sub.33H.sub.46ClN.sub.9O.sub.7+H].sup.+ 716; HPLC, AUC=98.21%; t.sub.R=8.66 min, Method A.

Example 137

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(3-oxo-3-((2-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)(3-phenylpropyl)amino)ethyl)amino)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-137)

##STR00499##

[0851] Preparation of Exp-137a: To a solution of Int-34 (350 mg, 0.596 mmol) in MeOH (10 mL) was added D-glucose (107 mg, 0.596 mmol) followed by AcOH (42.9 mg, 0.715 mmol) and NaCNBH.sub.3 (44.9 mg, 0.715 mmol). The resulting solution was stirred at 50 C. for 6 h then 3-phenyl propanal (120 mg, 0.893 mmol), AcOH (53.5 mg, 0.893 mmol) and NaCNBH.sub.3 (57.1 mg, 0.893 mmol) were added and the solution was stirred at 50 C. for 1 h. The solvent was removed, and satd. NaHCO.sub.3 (10 mL) was added. The solid was filtered, washed with water (10 mL), azeotroped with MeOH (10 mL2) and 1 N HCl in MeOH to remove borane. The resulting residue was washed with MeOH/MTBE/hexanes to remove phenyl propyl aldehyde and alcohol then dried to afford Exp-137a (416 mg, 88%) as a white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.47 (m, 4H), 7.33-7.18 (m, 14H), 5.05 (s, 2H), 4.10-3.90 (m, 1H), 3.80-3.43 (m, 11H), 3.20-2.92 (m, 6H), 2.62-2.56 (m, 6H), 2.03 (br s, 2H), 1.65-1.52 (m, 4H); ESI (m/z) [C.sub.44H.sub.57N.sub.3O.sub.8+H].sup.+ 756.

[0852] Preparation of Exp-137b: To a solution of Exp-137a (50 mg, 0.063 mmol) in EtOH (3.0 mL) and water (1.0 mL) was added 20% Pd(OH).sub.2 (10 mg) and AcOH (26.7 mg, 0.445 mmol). The resultant mixture was stirred under hydrogen (balloon) at 50 C. for 6 h. After filtration, the solvent was removed, to afford compound Exp-137b (43 mg, 92%) as colorless syrup: ESI (m/z) [C.sub.36H.sub.51N.sub.3O.sub.6+H].sup.+ 622.

[0853] Preparation of Exp-137: To a solution of compound Exp-137b (300 mg, 0.404 mmol) in DMF (6.0 mL) and EtOH (3.0 mL) was added compound 1 (157 mg, 0.404 mmol) at room temperature. The resultant mixture was stirred at 60 C. for 6 h. After the solvent was removed, the residue was purified by reverse phase column (5% to 80% of CH.sub.3CN in water, product came out at 40%), to afford compound Exp-137 (213 mg, 63%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.29-7.21 (m, 9H), 4.07-4.00 (m, 1H), 3.81-3.63 (m, 5H), 3.50-3.40 (m, 4H), 3.37-3.16 (m, 6H), 2.95 (t, J=7.5 Hz, 2H), 2.75-2.68 (m, 4H), 2.57 (t, J=7.5 Hz, 2H), 2.03-2.02 (m, 2H), 1.79-1.74 (m, 4H); ESI (m/z) [C.sub.42H.sub.56ClN.sub.9O.sub.7+H].sup.+ 834; HPLC, AUC=97.2%; t.sub.R=9.50 min, Method A.

Example 138

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(3-((2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-138)

##STR00500##

[0854] Preparation of Exp-138: In a three-step procedure Exp-138 was synthesized beginning with Int-34 and n-hexanal using a sequence analogous to the one used to prepare Exp-137: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.51 (m, 4H), 7.29-7.27 (m, 4H), 4.13-4.10 (m, 1H), 3.82-3.48 (m, 5H), 3.50-3.16 (m, 10H), 2.96 (t, J=7.5 Hz, 2H), 2.73 (t, J=7.0 Hz, 2H), 2.58 (t, J=7.5 Hz, 2H), 1.79-1.72 (m, 6H), 1.34-1.30 (m, 6H), 0.92-0.89 (m, 3H); ESI (m/z) [C.sub.39H.sub.58ClN.sub.9O.sub.7+H].sup.+ 800; HPLC, AUC=97.8%; t.sub.R=9.22 min, Method A.

Example 139

Preparation of (2R,2R,3R,3R,4S,4'S,5S,5'S,6S,6'S)-6,6-(((2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)ethyl)azanediyl)bis (ethane-2,1-diyl))bis(3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid) (Exp-139)

##STR00501## ##STR00502##

[0855] Preparation of Exp-139a: To a stirred solution of compound Int-34 (350 mg, 0.73 mmol) and compound Int-30 (799 mg, 2.22 mmol) in MeOH (7.0 mL) was added AcOH (0.1 mL, 1.46 mmol) and stirred for 15 min. The reaction was cooled to 0 C., NaCNBH.sub.3 (138 mg, 2.22 mmol) was added and resulted mixture was warmed to ambient temperature and stirring continued for 15 h. The volatiles were removed under reduced pressure and the obtained crude was diluted with CH.sub.2Cl.sub.2 (30.0 mL) and water (30.0 mL). The organic layer was separated, and the aqueous layer was again washed with CH.sub.2Cl.sub.2 (30.0 mL). The combined organic layer was washed with brine solution (50.0 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and evaporated under reduced pressure. The obtained crude product was purified by flash column chromatography (silica gel 12 g column) using 5% MeOH in CH.sub.2Cl.sub.2 to afford compound Exp-139a (700 mg, 75%) as an off-white solid. ESI (m/z) [C.sub.59H.sub.75N.sub.3O.sub.21+H].sup.+ 1162.

[0856] Preparation of Exp-139b: To a stirred solution of Exp-139a (650 mg, 0.56 mmol) in IPA/water (11.0 mL, 10:1 ratio) was added 20% Pd(OH).sub.2 on carbon 50% wet (160 mg, 25% weight substrate) at room temperature under nitrogen atmosphere. The nitrogen atmosphere was replaced with H.sub.2 (balloon) and the reaction mixture was stirred for 15 h. The reaction mixture was filtered through celite pad and the solids were washed with IPA and water (20.0 mL, 1:1 ratio). The filtrate was concentrated under reduced pressure to afford Exp-139b (550 mg, 95%) as an off-white sticky solid. ESI (m/z) [C.sub.51H.sub.469N.sub.3O.sub.19+H].sup.+ 1028.

[0857] Preparation of Exp-139c: To a stirred solution of Exp-139b (550 mg, 0.53 mmol) in DMF (5.0 mL) were added compound 1 (139 mg, 0.53 mmol) and DIPEA (0.6 mL, 3.21 mmol) at room temperature under nitrogen atmosphere. The reaction mixture was stirred at 50-60 C. for 15 h. The reaction mixture was cooled, and volatiles were removed by reduced pressure. The obtained crude product was purified by C-18 reverse phase column chromatography using 60-75% CH.sub.3CN in H.sub.2O to afford Exp-139c (330 mg, 50%) as a light-yellow solid. ESI (m/z) [C.sub.57H.sub.74ClN.sub.9O.sub.24+H].sup.+ 1240.

[0858] Preparation of Exp-139d: To a stirred solution of Exp-139c (330 mg, 0.27 mmol) in MeOH (10.0 mL) was added K.sub.2CO.sub.3 (73 mg, 0.53 mmol) at 0 C. The reaction mixture was warmed to ambient temperature and stirring was continued for 15 h. The volatiles were removed under reduced pressure to afford Exp-139d (300 mg, crude) as a white sticky solid. ESI (m/z) [C.sub.45H.sub.62ClN.sub.9O.sub.14+H].sup.+ 988.

[0859] Preparation of Exp 139: To a stirred solution of Exp-139d (330 mg crude, 0.33 mmol) in MeOH/THF/water (5.0 mL, 1:1:0.5 ratio) was added LiOH.Math.H.sub.2O (28 mg, 0.66 mmol) at 0 C. The reaction mixture was warmed to ambient temperature and stirring was continued for 5 h. The volatiles were removed under reduced pressure and the resulting residue was diluted with water (8.0 mL). The pH of the solution was adjusted to 4 with 2.0N aqueous HCl then the volatiles were removed under reduced pressure. The crude product was purified by reverse phase (C-18) column chromatography using 45% CH.sub.3CN in water; 0.1% HCl as a buffer to afford Exp-139 (75 mg, 14% after two steps) as a light-yellow powder. .sup.1H NMR (400 MHz, MeOH-d.sub.4) 7.42 (d, J=7.2 Hz, 2H), 7.18 (d, J=6.8 Hz 2H), 4.35-4.31 (m, 3H), 4.09-4.01 (m, 1H), 3.90-3.84 (m, 1H), 3.81-3.75 (m, 1H), 3.59-3.48 (m, 4H), 3.46-3.42 (m, 6H), 3.33-3.30 (m, 2H), 3.22-3.13 (m, 2H), 2.85 (t, J=7.6 Hz, 2H), 2.63 (t, J=6.8 Hz, 2H), 2.64 (t, J=7.6 Hz, 2H), 2.22-2.12 (m, 2H), 1.86-1.65 (m, 6H); ESI (m/z) [C.sub.43H.sub.58ClN.sub.9O.sub.14+H].sup.+ 960; HPLC, AUC=99.7%; t.sub.R=8.93 min, Method M.

Example 140

Preparation of 3,5-diamino-N(N-(4-(4-(3-((2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxy hexyl)amino)ethyl)(methyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-140)

##STR00503##

[0860] Preparation of Exp-140a: To a stirred solution of Int-23 (1.30 g, 3.00 mmol) in DMF (25 mL) was added DIPEA (1.1 g, 9.00 mmol), followed by the addition of HATU (1.70 g, 4.50 mmol). Upon stirring for 10 min, tert-butyl (2-(methylamino)ethyl)carbamate (0.62 g, 3.60 mmol) was added and stirring was continued for 4 h under N.sub.2 atmosphere at ambient temperature. The reaction mixture was diluted with brine solution (50 mL) and extracted with EtOAc (250 mL). The combined organic layers were washed with brine solution (50 mL), dried over anhydrous Na.sub.2SO.sub.4 and evaporated under reduced pressure. The obtained crude compound was purified by combi-flash column chromatography (silica gel 40 g column) using 5% MeOH in dichloromethane to afford Exp-140a (1.40 g, 78%) as an off-white solid. ESI (m/z) [C.sub.35H.sub.45N.sub.3O.sub.5+H].sup.+ 588.

[0861] Preparation of Exp-140b: To a stirred solution of Exp-140a (1.40 g, 2.47 mmol) in dichloromethane (5.0 mL) was added to 4.0 M HCl in 1,4-dioxane (6.0 mL) at 0 C. and stirring was continued at ambient temperature for 3 h. Then, the volatiles were removed under reduced pressure to afford Exp-140b (1.30 g, crude) as a white solid. ESI (m/z) [C.sub.30H.sub.37N.sub.3O.sub.3+H].sup.+ 488.

[0862] Preparation of Exp-140c: To a stirred solution of Exp-140b (1.30 g, 2.60 mmol) and D-glucose (1.40 g, 8.00 mmol) in MeOH (60.0 mL) was added AcOH (0.48 g, 8.00 mmol). The reaction mixture was stirred for 15 min and cooled to 0 C. NaCNBH.sub.3 (0.50 g, 8.00 mmol) was added and the resultant reaction mixture was warmed to 50 C. then stirred for 15 h. The reaction mixture was cooled, and the volatiles were removed under reduced pressure. The obtained crude material was purified by C-18 reverse phase column chromatography using 40-50% CH.sub.3CN in H.sub.2O to afford Exp-140c (1.10 g, 46%) as a colourless solid. ESI (m/z) [C.sub.42H.sub.61N.sub.3O.sub.13+H].sup.+ 816.

[0863] Preparation of Exp-140d: To a stirred solution of Exp-140c (1.0 g, 1.20 mmol) in IPA/Water (20.0 mL, 7:3) was added 20% Pd(OH).sub.2 on carbon 50% wet (0.20 g, 25% weight substrate) at room temperature under nitrogen atmosphere. Then, nitrogen was replaced with H.sub.2 (balloon) and stirred for 15 h. The reaction mixture was filtered through celite pad and the solids were washed with IPA and water (50 mL, 1:1). The combined filtrate was concentrated under reduced pressure to afford Exp-140d (0.75 g, 90%) as an off-white sticky solid. ESI (m/z) [C.sub.34H.sub.55N.sub.3O.sub.11+H].sup.+ 682.

[0864] Preparation of Exp-140: To a stirred solution of Exp-140d (0.70 g, 1.00 mmol) in DMF (15.0 mL) was added compound 1 (0.29 g, 1.10 mmol) and DIPEA (0.78 g, 6.00 mmol) at room temperature under nitrogen atmosphere. The reaction mixture was stirred at 50 C. for 15 h. The reaction mixture was cooled, and the volatiles were removed by reduced pressure. The obtained crude was purified by C-18 reverse phase column chromatography using 0-26% CH.sub.3CN in H.sub.2O to afford compound Exp-140 (0.20 g, 21%) as a yellow solid. .sup.1H NMR (400 MHz, CD.sub.3OD) 7.60-7.53 (m, 4H), 7.37-7.28 (m, 4H), 3.75-3.70 (m, 2H), 3.68-3.61 (m, 3H), 3.59-3.53 (m, 3H), 3.52-3.44 (m, 5H), 3.43-3.26 (m, 7H), 3.01 (s, 3H), 2.94-2.81 (m, 4H), 2.77-2.64 (m, 4H), 1.75-1.57 (m, 4H); ESI (m/z) [C.sub.40H.sub.60ClN.sub.9O.sub.12+H].sup.+ 894; HPLC, AUC=99.0%; t.sub.R=6.21 min, Method J.

Example 141

Preparation 3,5-diamino-N(N-(4-(4-(3-((2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxy hexyl)amino)ethyl)amino)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-141)

##STR00504##

[0865] Preparation of Exp-141a: To a solution of Int-16 (100 mg, 0.239 mmol) in DCM (3.0 mL) was added Dess-Martin reagent (122 mg, 0.287 mmol) at 0 C. The resultant mixture was stirred at 0 C. for 2 h and at room temperature for 2 h. The solvent was removed and to the residue was added DCM/MTBE (5.0 mL/5.0 mL). The solid was filtered out and washed with DCM. The combined filtrates were concentrated, partitioned with DCM and satd. Na.sub.2CO.sub.3 (10 mL/10 mL). The water layer was extracted with DCM (10 mL2), and the combined DCM layer was concentrated, to afford compound Exp-141a (95 mg, 95%) as a yellow solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 9.85 (s, 1H), 7.51-7.20 (m, 13H), 5.09 (s, 2H), 4.70 (br s, 1H), 3.26-3.24 (m, 2H), 3.02-2.97 (m, 2H), 2.84-2.79 (m, 2H), 2.68-2.63 (m, 2H), 1.71-1.67 (m, 4H).

[0866] Preparation of Exp-141b: To a suspension of Exp-141a (90 mg, 0.217 mmol), Int-20 (138 mg, 0.325 mmol) in DMF (3.0 mL) was added Pic-BH.sub.3 (34.8 mg, 0.325 mmol). The reaction mixture was stirred at 40 C. for 3 h and 50 C. for 3 h. After the solvent was removed, the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in water, product came out at 30% to 40%). The pure fractions were collected, concentrated and azeotroped with 1 N HCl in MeOH, to afford Exp-141b (89 mg, 47%) as a colorless syrup: ESI (m/z) [C.sub.41H.sub.61N.sub.3O.sub.12+H].sup.+ 788.

[0867] Preparation of Exp-141c: To a solution of Exp-141b (100 mg, 0.116 mmol) in EtOH (1.0 mL) and water (2.0 mL) was added 20% Pd(OH).sub.2 (15 mg). The resultant mixture was stirred under hydrogen (balloon) at 45 C. for 4 h. After filtration, fresh 20% Pd(OH).sub.2 (15 mg) and AcOH (0.1 mL) was added. The resultant suspension was stirred at 50 C. under hydrogen for 16 h. After filtration, fresh 20% Pd(OH).sub.2 (15 mg) was added. The resulting suspension was stirred at 60 C. under hydrogen for 8 h. After filtration, the solvent was removed, to afford Exp-141c (83 mg, 86%) as colorless oil: .sup.1H NMR (500 MHz, D.sub.2O) 7.58-7.54 (m, 4H), 7.31-7.28 (m, 4H), 4.09 (br s, 2H), 3.73-2.61 (m, 28H), 2.01 (s, 10H), 1.63-1.60 (m, 4H); ESI (m/z) [C.sub.33H.sub.55N.sub.3O.sub.10+H].sup.+ 654.

[0868] Preparation of Exp-141: To a solution of Exp-141c (278 mg, 0.333 mmol) in DMF (9.0 mL) was added compound 1 (130 mg, 0.333 mmol) and DIPEA (345 mg, 2.67 mmol) at room temperature. The resultant mixture was stirred at 65 C. for 16 h. After the solvent was removed, the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in water, product came out at 30% to 50%), to afford Exp-141 (231 mg, 80%) as a yellow solid: .sup.1H NMR (500 MHz, D.sub.2O) 7.53 (d, J=8.0 Hz, 2H), 7.44 (d, J=8.0 Hz, 2H), 7.30 (d, J=8.0 Hz, 2H), 7.20 (d, J=8.0 Hz, 2H), 3.85 (br s, 2H), 3.75-3.55 (m, 10H), 3.18-2.59 (m, 16H), 2.00-1.97 (m, 2H), 1.78-1.65 (m, 4H); ESI (m/z) [C.sub.39H.sub.60ClN.sub.9O.sub.11+H].sup.+ 866; HPLC, AUC=98.3%; t.sub.R=8.15 min, Method A.

Example 142

Preparation of 3,5-diamino-N(N-(4-(4-(6-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxy hexyl)amino)hexyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-142)

##STR00505##

[0869] Preparation of Exp-142a: To a solution of Int-42 (66.0 mg, 0.144 mmol) in MeOH (3.0 mL) was added D-glucose (78.0 mg, 0.432 mmol) followed by AcOH (25.9 mg, 0.432 mmol) and NaCNBH.sub.3 (27.1 mg, 0.432 mmol). The resulting solution was stirred at 60 C. for 6 h. Additional D-glucose (78.0 mg, 0.432 mmol), AcOH (25.9 mg, 0.432 mmol) and NaCNBH.sub.3 (27.1 mg, 0.432 mmol) were added. The resulting solution was stirred at 60 C. for 16 h. The solvent was removed then satd. Na.sub.2CO.sub.3 (3.0 mL) and MeOH (0.5 mL) were added. The resulting solid was filtered out and washed with water (1.0 mL). The solid was suspended in MeOH (5.0 mL) and 3N HCl in MeOH (0.5 mL) was added. After stirred at 40 C. for 10 min, a clear solution was obtained. After the solvent was removed, the residue was azeotroped with MeOH (5.0 mL) and 3N HCl in MeOH (20.5 mL) to afford compound Exp-142a (88 mg, 74%) as yellow syrup: ESI (m/z) [C.sub.42H.sub.62N.sub.2O.sub.12+H].sup.+ 785.

[0870] Preparation of Exp-142b: To a solution of Exp-142a (87 mg, 0.106 mmol) in EtOH (1.0 mL) and water (2.0 mL) was added 20% of Pd(OH).sub.2/C (10 mg). The resultant mixture was stirred under hydrogen (balloon) at 50 C. for 6 h. After filtration, the solvent was removed, to afford Exp-142b (75 mg, 95%) as brown syrup: ESI (m/z) [C.sub.34H.sub.56N.sub.2O.sub.10+H].sup.+ 653.

[0871] Preparation of Exp-142: To a solution of Exp-142b (700 mg, 0.934 mmol) in DMF (20 mL) was added compound 1 (363 mg, 0.934 mmol) and DIPEA (724 mg, 5.60 mmol) at room temperature. The resultant mixture was stirred at 65 C. for 1 h. Some solid did not go into solution. The solvent was removed by rotovap, the residue was re-dissolved in DMF (20 mL) and EtOH (10 mL), a clear yellow solution was observed. The reaction mixture was heated at 65 C. for 3 h and 70 C. for 4 h. After the solvent removed, the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in water, product came out at 30% to 50%), to afford Exp-142 (550 mg, 68%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.47 (m, 4H), 7.28-7.23 (m, 4H), 4.17 (br s, 2H), 3.83-3.63 (m, 10H), 3.40-3.30 (m, 8H), 2.74-2.64 (m, 4H), 1.81-1.68 (m, 8H), 1.45 (br s, 4H); ESI (m/z) [C.sub.40H.sub.61ClN.sub.8O.sub.11+H].sup.+ 865; HPLC, AUC=96.6%; t.sub.R=9.13 min, Method A.

Example 143

Preparation of 3,5-diamino-N(N-(4-(4-(3-(((S)-6-amino-1-((2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)amino)-1-oxohexan-2-yl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-143)

##STR00506##

[0872] Preparation of Exp-143a: To a solution of Int-35a (350 mg, 0.519 mmol) in THF (3.0 mL), MeOH (3.0 mL) and water (1.0 mL) was added NaOH (207 mg, 5.19 mmol) at room temperature. The resultant mixture was stirred at room temperature for 3 h. After the solvent was removed, water (10 mL) was added. The pH was adjusted to 4 with 1 N HCl and solid precipitation was observed. The solid was collected by filtration, washed with water and dried in oven, to afford Exp-143a (320 mg, 93%) as a yellow solid: ESI (m/z) [C.sub.38H.sub.49N.sub.3O.sub.7+H].sup.+ 660.

[0873] Preparation of Exp-143b: To a solution of acid Exp-143a (300 mg, 0.455 mmol), amine Int-20 (232 mg, 0.546 mmol) and DIPEA (176 mg, 1.364 mmol) in DMF (1.0 mL) was added HATU (207 mg, 0.546 mmol). The reaction mixture was stirred at room temperature for 2 h. After the solvent was removed, the residue was purified by reverse phase column (5% to 90% of CH.sub.3CN in H.sub.2O, product came out at 80%), to afford Exp-143b (376 mg, 80%) as a white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.33-7.22 (m, 9H), 5.05 (s, 2H), 4.25-4.20 (m, 1H), 3.84-3.62 (m, 12H), 3.50-3.40 (m, 6H), 3.16-3.13 (m, 2H), 2.97-2.94 (m, 4H), 2.69-2.59 (m, 6H), 1.66-1.53 (m, 6H), 1.41 (m, 9H), 1.40-1.21 (m, 4H).

[0874] Preparation of Exp-143c: To a solution of Exp-143b (374 mg, 0.363 mmol) in MeOH (12 mL) was added 20% Pd(OH).sub.2 (120 mg). The resultant mixture was stirred under hydrogen (balloon) at room temperature for 16 h. After filtration, the solvent was removed, to afford Exp-143c (295 mg, 91%) as colorless oil: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.50 (m, 4H), 7.30-7.25 (m, 4H), 4.25-4.22 (m, 1H), 3.83-3.53 (m, 12H), 3.50-3.40 (m, 2H), 3.19-2.60 (m, 12H), 2.97-2.94 (m, 4H), 1.74-1.53 (m, 6H), 1.41 (m, 9H), 1.40-1.21 (m, 4H); ESI (m/z) [C.sub.44H.sub.73N.sub.5O.sub.14+H].sup.+ 896.

[0875] Preparation of Exp-143d: To a solution of Exp-143c (50.0 mg, 0.056 mmol) in EtOH (1.0 mL) and DMF (1.0 mL) was added compound 1 (23.8 mg, 0.061 mmol) at room temperature. The resultant mixture was stirred under nitrogen at 70 C. for 8 h. After the solvent was removed, EtOH (1.0 mL) and IPA (1.0 mL) were added. The precipitated solids were filtered and dried to afford Exp-143d (52 mg, 84%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.49 (m, 4H), 7.29-7.26 (m, 4H), 4.24-4.19 (m, 1H), 3.96-3.53 (m, 16H), 3.50-3.40 (m, 2H), 3.06-2.58 (m, 12H), 1.75-1.53 (m, 6H), 1.41 (m, 9H), 1.40-1.21 (m, 4H).

[0876] Preparation of Exp-143: To a solution of Exp-143d (258 mg, 0.233 mmol) in EtOH (5.0 mL) was added 4 N HCl (10 mL) at room temperature. The resultant mixture was stirred at room temperature for 4 h. After the solvent was removed, the residue was re-dissolved in water (2.0 mL) and EtOH (10 mL) added. Solid precipitation was observed. The solvent was decanted and the solid was dried, to afford Exp-143 (239 mg, 87%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.51 (m, 4H), 7.30-7.28 (m, 4H), 4.30-4.10 (m, 3H), 3.85-3.36 (m, 20H), 2.98-2.89 (m, 4H), 2.73 (t, J=7.5 Hz, 2H), 2.64 (t, J=7.5 Hz, 2H), 1.79-1.30 (m, 10H); ESI (m/z) [C.sub.45H.sub.70ClN.sub.11O.sub.13+H].sup.2+/2 504; HPLC, AUC=94.6%; t.sub.R=8.36 min, Method A.

Example 144

Preparation of 3,5-diamino-N(N-(4-(4-(3-((S)-2-((2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)carbamoyl)pyrrolidin-1-yl)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-144)

##STR00507##

[0877] Preparation of Exp-144a: To a stirred solution of Int-23 (1.50 g, 3.40 mmol) in DMF (15.0 mL) was added DIPEA (0.90 g, 6.96 mmol) followed by HATU (1.32 g, 3.48 mmol), The reaction mixture was stirred for 15 mins, (L)-proline methyl ester (580 mg, 3.48 mmol) was added followed by stirring at rt for 6 h. The reaction mixture was diluted with water (220 mL) and extracted with EtOAc (3150 mL). The combined organic layer was washed with brine solution (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure to afford crude. The obtained crude was purified by combi-flash chromatography eluted with (70% EtOAc in hexanes) to afford Exp-144a (1.50 g, 80%) as an off-white solid. ESI (m/z) [C.sub.33H.sub.38N.sub.2O.sub.5+H].sup.+ 543.

[0878] Preparation of Exp-144b: To a stirred solution of Exp-144a (1.60 g, 2.95 mmol) in THF (15 mL) and H.sub.2O (4 mL), was added LiOH.Math.H.sub.2O (247 mg, 5.90 mmol) at room temperature. The reaction mixture was stirred at room temperature for 5 h then the reaction solvent was concentrated under reduced pressure. The resulting residue was taken into water (10 mL) and neutralized with 2 N HCl (4 mL). The resulting solid was filtered, washed with water (10 mL) and dried to give Exp-144b (1.45 g, 96%) as a white solid. ESI (m/z) [C.sub.32H.sub.36N.sub.2O.sub.5+H].sup.+ 529

[0879] Preparation of Exp-144c: To a stirred solution of Exp-144b (1.45 g, 2.74 mmol) in DMF (25.0 mL) was added DIPEA (700 mg, 5.48 mmol) followed by HATU (1.25 g, 3.29 mmol). The reaction mixture was stirred for 15 mins, compound Int-20 (1.06 g, 2.74 mmol) was added, and the reaction mixture stirred at rt for 6 h. After completion of the reaction, the reaction mixture was completely concentrated under reduced pressure and residue was washed with MTBE followed by reverse phase purification using 40% acetonitrile and water to afford Exp-144c (900 mg, 36%) as off-white gummy solid. ESI (m/z) [C.sub.46H.sub.66N.sub.4O.sub.14+H].sup.+ 899.

[0880] Preparation of Exp-144d: To a stirred solution of Exp-144c (800 mg, 0.88 mmol) in a mixture of IPA (15.0 mL), water (2 mL) and AcOH (1 mL) was charged with 20% Pd(OH).sub.2 on carbon 50% wet (100 mg, 50% weight substrate). The reaction mixture was stirred under H.sub.2 (balloon) for 12 h. The reaction mixture was filtered through celite and filtrate was concentrated under reduced pressure. The obtained crude was purified by reverse phase chromatography using 40% acetonitrile and water to afford Exp-144d (500 mg, 68%) as an off-white gummy solid. ESI (m/z) [C.sub.38H.sub.60N.sub.4O.sub.12+H].sup.+ 765.

[0881] Preparation of Exp-144: To a stirred solution of Exp-144d (500 mg, 0.65 mmol) and compound 1 (249 mg, 0.65 mmol) in DMF (20.0 mL), was charged DIPEA (500 mg, 3.92 mmol) at room temperature. The resultant reaction mixture was heated to 65 C. and stirred for 6 h. The reaction mixture was concentrated and washed with CH.sub.2Cl.sub.2 (20 mL) followed by reverse phase purification using 40% acetonitrile and water to afford Exp-144 (134 mg, 21%) as yellowish brown solid: .sup.1H NMR (400 MHz, DMSO-d.sub.6, D.sub.2O exchange) 7.59-7.50 (m, 4H), 7.36-7.23 (m, 4H), 4.31-4.18 (m, 1H), 3.85-3.68 (m, 2H), 3.67-3.57 (m, 4H), 3.57-3.52 (m, 3H) 3.49-3.39 (m, 7H) 3.39-3.26 (m, 3H) 3.17-3.11 (m, 1H) 2.92-2.80 (m, 3H) 2.73-2.59 (m, 6H) 2.34-2.14 (m, 1H) 2.10-1.97 (m, 1H), 1.92-1.79 (m, 3H), 1.70-1.56 (m, 3H) 1.50-1.40 (m, 1H); ESI (m/z) [C.sub.44H.sub.65ClN.sub.10O.sub.13+H].sup.+ 977; HPLC, AUC=97.7%; t.sub.R=6.26 min, Method J.

Example 145

Preparation of (R)-1-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino) butyl)-[1,1-biphenyl]-4-yl)propanoyl)piperazine-2-carboxylic acid (Exp-145)

##STR00508##

[0882] Preparation of Exp-145a: To a solution of Int-23 (1.40 g, 3.24 mmol) in DMF (30 mL) was added 1-(tert-butyl) 3-methyl (R)-piperazine-1,3-dicarboxylate (793 mg, 3.23 mmol), HATU (1.48 g, 3.89 mmol) and DIPEA (1.25 g, 9.73 mmol). The resulting solution was stirred at rt for 1 h. The reaction mixture was partitioned with water (150 mL) and EtOAc (150 mL) then separated. The water layer was washed with EtOAc (150 mL). The organic layers were combined, dried and concentrated, to afford Exp-145a (2.02 g, 95%) as a white solid: ESI (m/z) [C.sub.38H.sub.47N.sub.3O.sub.7+H].sup.+ 658.

[0883] Preparation of Exp-145b: To a solution of Exp-145a (970 mg, 1.47 mmol) in IPA/MeOH/H.sub.2O (10 mL/10 mL/10 mL) was added Pd(OH).sub.2/C (20% on carbon, 120 mg, 50% wet) under nitrogen. The suspension was stirred at 45 C. for 2 h with a continuous hydrogen purge. After filtration, fresh Pd(OH).sub.2/C (20% on carbon, 120 mg, 50% wet) was added. The suspension was stirred at 45 C. for 2 h under hydrogen. After filtration, the solvent was removed, to afford Exp-145b (810 mg, 94%) as a white solid: ESI (m/z) [C.sub.30H.sub.41N.sub.3O.sub.5+H].sup.+ 524.

[0884] Preparation of Exp-145c: To a solution of compound Exp-145b (810 mg, 1.38 mmol) in DMF (10 mL) and EtOH (10 mL) was added compound 1 (539 mg, 1.38 mmol) and DIPEA (897 mg, 6.94 mmol). The resulting solution was stirred at 55 C. for 16 hr. After the solvent was removed, water (30 mL) was added. The solid was filtered and dried, to afford Exp-145c (938 mg, 92%) as a yellow solid: ESI (m/z) [C.sub.36H.sub.46ClN.sub.9O.sub.6+H].sup.+ 736.

[0885] Preparation of Exp-145d: To a solution of Exp-145c (938 mg, 1.27 mmol) in MeOH (5.0 mL) was added 3 N HCl in MeOH (10 mL). The resulting solution was stirred at rt for 4 hr. After the solvent was removed, the residue was purified by reverse phase column (10% to 90% of CH.sub.3CN in water, product came out at 40% to 80%), to afford compound Exp-145d (482 mg, 53%, contaminated with 10% of ethyl ester) as a yellow solid.

[0886] Preparation of Exp-145: To a suspension of Exp-145d (180 mg, 0.254 mmol) in THF/MeOH/H.sub.2O (4.0 mL/4.0 mL/4.0 mL) was added NaOH (102 mg, 2.54 mmol). The reaction mixture was stirred at rt for 4 h. The pH of reaction mixture was adjusted to 6. The solvent was removed, and water (3 mL) was added. The solid was collected by filtration, to afford Exp-145 (80.0 mg, 63%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.46 (m, 4H), 7.31-7.26 (m, 4H), 5.02 (br s, 0.5H), 4.85 (br s, 0.5H), 4.39-4.37 (m, 0.5H), 4.30 (br s, 0.5H), 3.69-3.38 (m, 3H), 3.07-2.43 (m, 10H), 1.81-1.70 (m, 4H); ESI (m/z) [C.sub.30H.sub.36ClN.sub.9O.sub.4+H].sup.+ 622; HPLC, AUC=96.2%; t.sub.R=9.47 min, Method A.

Example 146

Preparation of methyl (R)-1-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl) guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)piperazine-2-carboxylate (Exp-146)

##STR00509##

[0887] Preparation Exp-146: To a solution of Exp-145 (220 mg, 0.317 mmol) in MeOH (15 mL) was added SOCl.sub.2 (0.5 mL). The reaction mixture was stirred at 35 C. for 16 h. After the solvent was removed, the residue was purified by reverse phase column (10% to 80% of CH.sub.3CN in water, product came out at 40%), to afford Exp-146 (86 mg, 38%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.51 (m, 4H), 7.32-7.27 (m, 4H), 5.55 (br s, 0.5H), 5.20 (br s, 0.5H), 4.70-4.65 (m, 0.5H), 4.14-4.13 (m, 0.5H), 3.90-3.67 (m, 5H), 3.44-3.16 (m, 3H), 3.02-2.71 (m, 8H), 1.81-1.72 (m, 4H); ESI (m/z) [C.sub.31H.sub.38ClN.sub.9O.sub.4+H].sup.+ 636; HPLC, AUC=95.5%; t.sub.R=9.72 min, Method A.

Example 147

Preparation of methyl (R)-4-(3-aminopropyl)-1-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)piperazine-2-carboxylate (Exp-147)

##STR00510##

[0888] Preparation of Exp-147a: To a solution of Exp-145a (1.50 g, 2.28 mmol) in MeOH (15 mL) was added 3 N HCl in MeOH (15 mL). The resulting solution was stirred at rt for 8 h. The solvent was removed, to afford compound Exp-147a (1.36 g, 100%) as a yellow oil: ESI (m/z) [C.sub.33H.sub.39N.sub.3O.sub.5+H].sup.+ 558.

[0889] Preparation of Exp-147b: To a solution of Exp-147a (900 mg, 1.61 mmol) in MeOH (30 mL) was added tert-butyl (3-oxopropyl)carbamate (419 mg, 2.42 mmol), AcOH (145 mg, 2.42 mmol) and NaCNBH.sub.3 (152 mg, 2.42 mmol). The resulting solution was stirred at 50 C. for 2 h. Additional tert-butyl (3-oxopropyl)carbamate (279 mg, 1.61 mmol), AcOH (96.6 mg, 1.61 mmol) and NaCNBH.sub.3 (101 mg, 1.61 mmol) were added. The resultant solution was stirred at 50 C. for 3 h. After the solvent was removed, the residue was partitioned with NaHCO.sub.3 (60 mL) and EtOAc (100 mL). The organic layer was collected, dried and concentrated, to afford Exp-147b (1.40 g, 121%, crude) as yellow solid: ESI (m/z) [C.sub.41H.sub.54N.sub.4O.sub.7+H].sup.+ 715.

[0890] Preparation of Exp-147c: To a solution of Exp-147b (600 mg, 0.839 mmol) in IPA/MeOH/H.sub.2O (6.0 mL/12 mL/6.0 mL) was added AcOH (3.02 mg, 5.04 mmol) and Pd(OH).sub.2/C (20% on carbon, 100 mg, 50% wet) under nitrogen. The suspension was stirred at 45 C. for 4 h with a continuous hydrogen purge. After filtration, fresh Pd(OH).sub.2/C (20% on carbon, 100 mg, 50% wet) was added. The suspension was stirred at 45 C. for 4 h under hydrogen. After filtration, the solvent was removed to afford Exp-147c (558 mg, 95%) as a white solid: ESI (m/z) [C.sub.33H.sub.48N.sub.4O.sub.5+H].sup.+ 581.

[0891] Preparation of Exp-147d: To a solution of Exp-147c (100 mg, 0.143 mmol) in DMF (3.0 mL) was added compound 1 (55.4 mg, 0.143 mmol) and DIPEA (55.3 mg, 0.428 mmol). The resulting solution was stirred at 55 C. for 8 hr. After the solvent was removed, water (10 mL) was added. The water phase was decanted, and the residue was azeotroped with MeOH (20 mL), to afford Exp-147d (104 mg, 92%, crude) as yellow oil: ESI (m/z) [C.sub.39H.sub.53ClN.sub.10O.sub.6+H].sup.+ 793.

[0892] Preparation of Exp-147: To a solution of Exp-147d (376 mg, 0.474 mmol) in MeOH (3.0 mL) was 3 N HCl in MeOH (6.0 mL). The resulting solution was stirred at rt for 4 hr. After the solvent was removed, the residue was purified by reverse phase column (10% to 90% of CH.sub.3CN in water, product came out at 40% to 80%), to afford Exp-147 (149 mg, 39%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.31-7.27 (m, 4H), 5.22 (br s, 1H), 3.86-3.83 (m, 1H), 3.75 (s, 3H), 3.43-3.34 (m, 4H), 3.02-2.80 (m, 4H), 2.76-2.70 (m, 5H), 2.61-2.42 (m, 2H), 2.12-1.72 (m, 8H); ESI (m/z) [C.sub.34H.sub.45ClN.sub.10O.sub.4+H].sup.+ 693; UPLC, AUC=97.5%; t.sub.R=3.10 min, Method B.

Example 148

Preparation of (R)-4-(3-aminopropyl)-1-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)piperazine-2-carboxylic acid (Exp-148)

##STR00511##

[0893] Preparation of Exp-148: To a solution of Exp-147 (102 mg, 0.147 mmol) in THF/MeOH/H.sub.2O (2.0 mL/2.0 mL/2.0 mL) was added NaOH (63.8 mg, 1.59 mmol). The resulting solution was stirred at rt for 4 hr. After pH was adjusted to 6 with 1 N HCl, the solvent was removed. The residue was purified by reverse phase column (10% to 90% of CH3CN in 0.01 M HCl, product came out at 40% to 80%), to afford Exp-148 (80.0 mg, 63%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.50 (m, 0.5H), 7.33-7.27 (m, 4H), 5.62 (br s, 0.5H), 5.28 (br s, 0.5H), 4.83-4.77 (m, 0.5H), 4.24-4.21 (m, 0.5H), 4.13-4.07 (m, 1H), 3.55-3.50 (m, 2H), 3.33-3.29 (m, 4H), 3.07-2.71 (m, 10H), 2.20-2.12 (m, 2H), 1.81-1.72 (m, 4H); ESI (m/z) [C.sub.33H.sub.43ClN.sub.10O.sub.4+H].sup.+ 679; UPLC, AUC=98.26%; t.sub.R=2.99 min, Method B.

Example 149

Preparation of methyl (R)-4-(3-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino) propyl)-1-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)piperazine-2-carboxylate (Exp-149)

##STR00512##

[0894] Preparation of Exp-149a: To a solution of Exp-147b (1.25 g, 1.958 mmol) in MeOH (10 mL) was added 3 N HCl in MeOH (10 mL). The resulting solution was stirred at rt for 4 h. The solvent was removed, to afford Exp-149a (1.35 g, 100%) as a yellow solid: ESI (m/z) [C.sub.36H.sub.46N.sub.4O.sub.5+H].sup.+ 615.

[0895] Preparation of Exp-149b: To a solution of Exp-149a (1.35 g, 2.19 mmol) in MeOH (30 mL) was added glucose (1.18 g, 6.59 mmol), AcOH (396 mg g, 6.59 mmol) and NaCNBH.sub.3 (414 mg, 6.59 mmol). The resultant solution was stirred at 50 C. for 16 h. Additional glucose (393 mg, 2.19 mmol), AcOH (131 mg, 2.19 mmol) and NaCNBH.sub.3 (137 mg, 2.19 mmol) were added and the resultant solution was stirred at 50 C. for 8 hr. Additional glucose (393 mg, 2.19 mmol), AcOH (131 mg, 2.19 mmol) and NaCNBH.sub.3 (137 mg, 2.19 mmol) were added and the resultant solution was stirred at 50 C. for 16 hr. After the solvent was removed, water (20 mL) and MeOH (10 mL) were added. The resulting solid was collected by filtration, washed with water (10 mL), azeotroped with MeOH (100 mL) to remove water, then azeotroped with 1 N HCl in MeOH (50 mL2) to remove boron, to afford 1.01 g of Exp-149b. The filtrate was purified by reverse phase column (10% to 80% of CH.sub.3CN in water, product came out at 40% to 80%; followed by 50% to 80% of CH.sub.3CN in 0.01 M HCl, more product washed out), the pure fractions were collected, concentrated and azeotroped with 1 N HCl in MeOH to remove boron, to afford 240 mg of compound Exp-149b. Total Exp-149b (1.25 g, 82%) was obtained as a yellow syrup: ESI (m/z) [C.sub.48H.sub.70N.sub.4O.sub.15+H].sup.+ 943.

[0896] Preparation of Exp-149c: To a solution of Exp-149b (1.01 g, 1.07 mmol) in IPA/MeOH/H.sub.2O (10 mL/20 mL/10 mL) was added AcOH (386 mg, 6.43 mmol) and Pd(OH).sub.2/C (20% on carbon, 100 mg, 50% wet). The suspension was stirred at 45 C. for 2 h with a continuous hydrogen purge. After filtration, fresh Pd(OH).sub.2/C (20% on carbon, 100 mg, 50% wet). The suspension was stirred at 45 C. for 2 h under hydrogen. After filtration, the solvent was removed, to afford Ep-149c (985 mg, 93%) as yellow oil: ESI (m/z) [C.sub.40H.sub.64N.sub.4O.sub.13+H].sup.+ 809.

[0897] Preparation of Exp-149: To a solution of Exp-149c (238 mg, 0.241 mmol) in DMF (5.0 mL) was added compound 1 (94.0 mg, 0.241 mmol) and DIPEA (155 mg, 1.20 mmol). The resulting solution was stirred at 55 C. for 8 hr. After the solvent was removed, the residue the residue was purified by reverse phase column (10% to 90% of CH.sub.3CN in water, product came out at 30% to 70%), to afford 46 mg of material as freebase. Some material stuck in column, which was washed with 30% of CH.sub.3CN in 0.01M HCl, to afford an additional 38 mg of material as HCl salt. Total Exp-149 (84 mg, 30%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.51 (m, 4H), 7.32-7.27 (m, 4H), 5.53 (br s, 0.5H), 4.63 (br s, 0.5H), 4.21-4.20 (m, 2H), 3.85-3.64 (m, 14H), 3.46-3.30 (m, 12H), 3.01-2.71 (m, 9H), 2.22 (br s, 2H), 1.81-1.72 (m, 4H); ESI (m/z) [C.sub.46H.sub.69ClN.sub.10O.sub.4+H].sup.+ 1022; HPLC, AUC=94.4%; t.sub.R=9.01 min, Method A.

Example 150

Preparation of (R)-4-(3-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino) propyl)-1-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl) propanoyl)piperazine-2-carboxylic acid (Exp-150)

##STR00513##

[0898] Preparation of Exp-150: To a solution of Exp-149 (369 mg, 0.361 mmol) in THF/MeOH/H.sub.2O (5.0 mL/5.0 mL/5.0 mL) was added NaOH (144 mg, 3.61 mmol). The resulting solution was stirred at rt for 4 hr. After the solvent was removed and water (10 mL) was added. The pH was adjusted to 2 with 1 N HCl. The solution was purified by reverse phase column (10% to 90% of CH3CN in 0.01 M HCl, product came out at 30% to 70%), to afford Exp-150 (212 mg, 52%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.51 (m, 4H), 7.33-7.27 (m, 4H), 5.51 (br s, 0.5H), 5.25 (br s, 0.5H), 4.23-4.20 (m, 4H), 3.87-3.37 (m, 22H), 3.36-2.72 (m, 8H), 2.40-2.30 (m, 2H), 1.81-1.72 (m, 4H); ESI (m/z) [C.sub.45H.sub.67ClN.sub.10O.sub.4+H].sup.+ 1007; HPLC, AUC=98.2%; t.sub.R=8.82 min, Method A.

Examples 151-154

Preparation of Exp-151-154: Examples 151-154 were prepared from Int-23 and 1-(tert-butyl) 3-methyl (S)-piperazine-1,3-dicarboxylate in an analogous sequence of procedures used for synthesis of Examples 147-150

##STR00514##

TABLE-US-00040 Exp # R.sup.0 R R.sup.1 Spectra 151 CH.sub.3 H H .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.50 (m, 4H), 7.31-7.27 (m, 4H), 5.35 (br s, 0.6H), 4.47 (br s, 0.3H), 3.97 (br s, 1H), 3.78-3.77 (m, 3H), 3.45-3.32 (m, 7H), 3.03-2.71 (m, 10H), 1.93-1.74 (m, 6 H); ESI (m/z) [C.sub.34H.sub.45ClN.sub.10O.sub.4 + H].sup.+ 693; HPLC, AUC = 99.5%; t.sub.R = 9.26 min, Method A. 152 H H H .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.50 (m, 4H), 7.33-7.27 (m, 4H), 5.61 (br s, 0.5H), 5.26 (br s, 0.4H), 4.72-4.69 (m, 0.5H), 4.24-4.21 (m, 0.6H), 4.12-4.07 (m, 1H), 3.56-3.50 (m, 1.5H), 3.37-3.20 (m, 5.5 H), 3.05-2.81 (m, 7H), 2.73 (t, J = 7.0 Hz, 2H), 2.19-2.13 (m, 2H), 1.95-1.72 (m, 4H); ESI (m/z) [C.sub.33H.sub.43ClN.sub.10O.sub.4 + H].sup.+ 679; HPLC, AUC = 96.8%; t.sub.R = 8.83 min, Method A. 153 CH.sub.3 Sug Sug .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.31-7.27 (m, 4H), 5.20 (br s, 0.7H), 4.30 (br s, 0.3H), 4.19-4.15 (m, 2H), 3.83-3.63 (m, 14H), 3.48-3.34 (m, 10H), 2.97-2.40 (m, 9H), 2.10-1.90 (m, 4H), 1.81-1.72 (m, 4H); ESI (m/z) [C.sub.46H.sub.69ClN.sub.10O.sub.14 + H].sup.+ 1021; HPLC, AUC = 99.0%; t.sub.R = 8.96 min, Method A. 154 H Sug Sug .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.50 (m, 4H), 7.32-7.27 (m, 4H), 5.52 (br s, 0.6H), 5.18 (br s, 0.4H), 4.21-4.20 (m, 2H), 4.15-4.13 (m, 2H), 3.86-3.64 (m, 10H), 3.60-3.30 (m, 10H), 3.20-2.71 (m, 10H), 2.25-2.23 (m, 2H), 1.79-1.74 (m, 4H); ESI (m/z) [C.sub.45H.sub.67ClN.sub.10O.sub.14 + H].sup.+ 1007; HPLC, AUC = 94.9%; t.sub.R = 8.77 min, Method A.

Example 155

Preparation of 3-amino-N(N-(4-(4-(3-((2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-5H-pyrrolo[2,3-b]pyrazine-2-carboxamide (Exp-155)

##STR00515##

[0899] Preparation of Exp-155a: To a stirred solution of Exp-87b (200 mg, 0.284 mmol) in DMF (5 mL) was added Int-44 (110 mg, 0.312 mmol) and DIPEA (74 mg, 0.568 mmol) at room temperature under nitrogen. The reaction mixture was stirred at 60 C. for 6 h. The reaction mixture was concentrated and the obtained crude product was purified by C-18 reverse phase column chromatography (5% CH.sub.3CN/0.01% AcOH in H.sub.2O) to afford compound Exp-155a (140 mg, 48%) as a brown solid; ESI (m/z) [C.sub.46H.sub.67N.sub.9O.sub.14+H].sup.+ 970.

[0900] Preparation of Exp-155: To a stirred solution of Exp-155a (140 mg, 0.144 mmol) in dichloromethane (5 mL) was added 4 M HCl in 1,4-dioxane (5 mL) at 0 C. The resulted mixture was warmed to ambient temperature and stirring continued for 4 h. After that, the reaction mixture was concentrated and twice purified by C-18 reverse phase column chromatography (5% CH.sub.3CN in H.sub.2O) to afford Int-155 (100 mg, 73%) as a pale brown solid. .sup.1H NMR (400 MHz, D.sub.2O) 7.60-7.58 (d, J=8.0 Hz, 1H), 7.48-7.42 (m, 2H), 7.34-7.29 (m, 1H), 7.28-7.21 (m, 3H), 6.93-6.91 (d, J=8.0 Hz, 2H), 6.52-6.51 (m, 1H), 4.15-4.04 (m, 2H), 3.76-3.65 (m, 7H), 3.61-3.48 (m, 5H), 3.42-3.31 (m, 5H), 3.29-3.26 (m, 1H), 3.25-3.22 (m, 1H), 3.21-3.17 (m, 2H), 2.79-2.71 (m, 1H), 2.66-2.57 (m, 2H), 2.53-2.45 (m, 2H), 1.79-1.59 (in, 4H); ESI (m/z) [C.sub.41H.sub.59N.sub.9O.sub.12+H].sup.+ 870; HPLC, AUC=93.26%; t.sub.R=8.84 min, Method N.

Examples 156-158

[0901] Preparation of Examples 156-158: In a scheme analogous to the four-step procedure used for preparation of Example 155, intermediates Int-31, 32 and 33 were coupled to Int-23 and moved forward to Examples 156-158.

TABLE-US-00041 Exp-# Structure Spectra 156 [00516] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.44-7.40 (m, 5H), 7.21-7.18 (m, 4H), 6.39-6.38 (d, 1H), 3.33-3.29 (t, J = 8 Hz, 2H), 2.88-2.84 (t, J = 12.0 Hz, 2H), 2.68-2.63 (m, 4H), 1.95-1.85 (m, 2H), 1.70-1.69 (m, 4H); ESI (m/z)) [C.sub.27H.sub.32N.sub.8OH].sup. 483; HPLC, AUC = 97.1%; t.sub.R = 6.35 min, Method J. Exp-155 157 [00517] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.44-7.40 (m, 5H), 7.21-7.15 (m, 4H), 6.37 (d, J = 4 Hz, 1H), 3.80-3.77 (m, J = 6.8 Hz, 1H), 3.32-3.26 (m, 2H), 3.17-3.14 (m, 2H), 2.86-2.82 (m, 2H), 2.67-2.65(m, 4H), 1.86-1.58(m, 10H), 1.44-1.31(m, 2H); ESI (m/z) [C.sub.33H.sub.44N.sub.10O.sub.2 + H].sup.+ 613; HPLC, AUC = 94.7%; t.sub.R = 6.29 min, Method J. Exp-156 158 [00518] .sup.1H NMR (400 MHz, CD.sub.3OD) 7.43-7.39 (m, 5 H), 7.20-7.15 (m, 4H), 6.39-6.37 (s, 1H), 4.10-4.06 (m, 2H), 3.83-3.80 (m, 1H), 3.68-3.66 (m, 2 H), 3.62-3.53 (m, 7H), 3.39- 3.29(m, 8H), 2.66-2.58 (m, 5H), 1.88-1.67 (m, 10H), 1.45- 1.40 (m, 2H), 1.24-1.20 (m, 2H), ESI (m/z) [C.sub.45H.sub.68N.sub.10O.sub.12 H].sup. 939; HPLC, AUC = 91.0%; t.sub.R = 8.87 min, Method N. Exp-157

Example 159

Preparation of (S)-3,5-diamino-N(N-(4-(4-(3-((5-amino-1-(1H-tetrazol-5-yl)pentyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-159)

##STR00519##

[0902] Preparation of Exp-159a: To a solution Int-50 (500 mg, 0.73 mmol) in IPA (20 mL) and EtOAc (20 mL), was added 20% Pd(OH).sub.2 on carbon 50% wet (200 mg, 50% weight substrate) under inert atmosphere. The resultant reaction mixture stirred under H.sub.2 (balloon pressure) for 16 h. The reaction mass filtered through celite bed and filtrate concentrated under reduced pressure to afford Exp-159a (380 mg, 97%) as an off-white solid; ESI (m/z) [C.sub.30H.sub.43N.sub.7O.sub.3+H].sup.+ 550.

[0903] Preparation of Exp-159b: A stirred solution of Exp-159a (380 mg, 0.69 mmol) and compound 1 (295 mg, 0.76 mmol) in DMF (10 mL), was charged with DIPEA (0.34 mL, 2.07 mmol) at room temperature. The resultant reaction mixture was heated to 60 C. and stirred for 8 h. The reaction mixture poured in a beaker containing cold water (20 mL), the precipitated solids were filtered and was washed with water (20 mL). The obtained solid was azeotroped with CH.sub.3CN (25 mL) and dried under reduced pressure to afford Exp-159b (400 mg, 75%) as a brown solid; ESI (m/z) [C.sub.36H.sub.48ClN.sub.13O.sub.4+H].sup.+ 762.

[0904] Preparation of Exp-159: A stirred solution of Exp-159b (350 mg, 0.46 mmol) in MeOH (5.0 mL) was charged 4M HCl in 1,4-dioxane (5.0 mL). The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure. The obtained crude material was purified by reverse phase combi-flash chromatography eluted with 10%-100% CH.sub.3CN in water (compound isolated at 36% of CH.sub.3CN) to afford Exp-159 (120 mg, 35%) as an off-white solid. .sup.1H NMR (400 MHz, CD.sub.3OD) 7.40 (d, J=7.6 Hz, 2H), 7.35 (d, J=8.0 Hz, 2H), 7.18-7.13 (m, 4H), 5.22-5.18 (m, 1H), 3.26-3.23 (m, 2H), 2.88-2.81 (m, 2H), 2.75 (t, J=7.6 Hz, 2H), 2.62 (t, J=6.8 Hz, 2H), 2.47 (t, J=7.6 Hz, 2H), 1.93-1.74 (m, 2H), 1.69-1.63 (m, 4H), 1.58-1.50 (m, 2H), 1.28-1.15 (m, 2H); ESI (m/z) [C.sub.31H.sub.40ClN.sub.13O.sub.2+H].sup.+ 662; HPLC, AUC=97.1%; t.sub.R=6.99 min, Method R.

Example 160

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(3-oxo-3-(((S)-5-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)-1-(1H-tetrazol-5-yl)pentyl)amino)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-160)

##STR00520##

[0905] Preparation of Int-160a: To a stirred solution of Int-50 (1.7 g, 2.48 mmol) in MeOH (5.0 mL) was slowly added 4M HCl in MeOH (10.0 mL) followed by stirring at room temperature for 5 h. The reaction mass was concentrated under reduced pressure, the resulting residue was washed with MTBE yielding a solid which was dried under reduced pressure to afford Exp-160a (1.45 g, 96%) as an off-white solid; ESI (m/z) [C.sub.33H.sub.41N.sub.7O.sub.3+H].sup.+ 584.

[0906] Preparation of Int-160b: To a stirred solution of compound Exp-160a (650 mg, 1.049 mmol) in MeOH (10 mL) was added D(+) glucose (189 mg, 1.049 mmol), AcOH (0.06 mL, 1.049 mmol) followed by NaBH.sub.3CN (65.9 mg, 1.049 mmol). The resultant reaction mixture was stirred at 45 C. for 24 h. The reaction mixture was cooled to room temperature and charged with D(+) glucose (94 mg, 0.524 mmol) followed by NaBH.sub.3CN (32.9 mg, 0.524 mmol) then continued stirring at 45 C. for another 36 h. The reaction mixture was concentrated under reduced pressure and crude material was purified by reverse phase combi flash purification eluted with 10-40% CH.sub.3CN in H.sub.2O (0.1% HCl as buffer) to afford Exp-160b (250 mg, 31%) as an off-white gummy solid: ESI (m/z) [C.sub.39H.sub.53N.sub.7O.sub.5+H].sup.+ 748.

[0907] Preparation of Int-160c: To a stirred solution of Exp-160b (250 mg, 0.334 mmol) in IPA (10 mL) and water (5 mL), EtOAc (10 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (100 mg, 50% weight substrate) under an inert atmosphere. The resultant reaction mixture was stirred under H.sub.2 (balloon) for 5 h. The reaction mixture was filtered through celite bed and the solids were washed with IPA:H.sub.2O (1:1). The combined filtrate was concentrated under reduced pressure. The obtained crude material was azeotroped with toluene (220 mL) and dried under reduced pressure to afford Exp-160c (200 mg, 97%) as an off-white gummy solid; ESI (m/z) [C.sub.31H.sub.47N.sub.7O.sub.6+H].sup.+ 614.

[0908] Preparation of Int-160: To a stirred solution of Exp-160c (200 mg, 0.325 mmol) and compound 1 (93.5 mg, 0.3589 mmol) in DMF (10 mL) was added DIPEA (0.3 mL, 1.625 mmol). The reaction mixture was warmed to 60 C. and stirred for 16 h. The reaction mixture was concentrated under reduced pressure and obtained crude material was dissolved in water (5 mL) and the pH=was adjusted to 2 with 2N HCl solution. The water was evaporated under reduced pressure and residue was purified by reverse phase combi flash column chromatography eluting with 10%-25% of ACN in H.sub.2O (0.01% HCl as buffer) (compound isolated at 20% of ACN) to afford Exp-160; .sup.1H NMR (400 MHz, CD.sub.3OD) 7.46-7.34 (m, 4H), 7.23-7.11 (m, 4H), 5.22-5.13 (m, 1H), 3.95-3.90 (m, 1H), 3.75-3.65 (m, 2H), 3.62-3.52 (m, 3H), 3.30-3.24 (m, 2H), 3.03-2.97 (m, 2H), 2.89-2.80 (m, 4H), 2.64 (t, J=6.92 Hz, 2H), 2.51 (t, J=7.48 Hz, 2H), 1.98-1.81 (m, 2H), 1.72-1.1.58 (m, 6H), 1.32-1.20 (m, 2H); ESI (m/z) [C.sub.37H.sub.52ClN.sub.13O.sub.7+H].sup.+ 826; HPLC AUC 97.8% (R.sub.t=7.61 min); HPLC, AUC=97.8%; t.sub.R=7.61 min, Method W.

Example 161

Preparation of 3,5-diamino-N(N-(4-(4-(3-(((S)-5-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)-1-(1H-tetrazol-5-yl)pentyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-161)

##STR00521##

[0909] Preparation of Exp-161a: To a stirred solution of Exp-160a (600 mg, 1.02 mmol) in MeOH (20 mL) was added D-(+) glucose (741 mg, 4.11 mmol), AcOH (0.18 mL, 3.08 mmoL) and NaBH.sub.3CN (258 mg, 4.11 mmol). The resultant reaction mixture was stirred at 60 C. for 24 h. The reaction mass was cooled to room temperature and charged with D-(+) glucose (370.8 mg, 2.05 mmol) followed by NaBH.sub.3CN (129.2 mg, 2.05 mmol) then continued stirring at 60 C. for another 36 h. The reaction mass concentrated under reduced pressure. The obtained crude material treated with 4N HCl in CH.sub.3OH (5 mL) stirred for 15 min and concentrated under reduced pressure. Crude material was purified by reverse phase combi flash purification eluting with 10-40% CH.sub.3CN in H.sub.2O (0.1% HCO.sub.2H as buffer) to afford Exp-161a (430 mg, 48%) as an off white gummy solid: ESI (m/z) [C.sub.45H.sub.65N.sub.7O.sub.13+H].sup.+ 912.

[0910] Preparation of Exp-161b: To a stirred solution of Exp-161a (430 mg, 0.47 mmol) in IPA (20 mL) and water (7 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (100 mg, 50% weight substrate) under inert atmosphere. The resultant reaction mixture stirred under H.sub.2 (balloon) for 6 h. The reaction mass filtered through a celite bed, and the filtrate was concentrated under reduced pressure. The obtained crude was azeotroped with toluene (220 mL) and dried under reduced pressure to afford Exp-161b (330 mg, 90%) as an off-white gummy solid; ESI (m/z) [C.sub.37H.sub.59N.sub.7O.sub.11+H].sup.+ 778.

[0911] Preparation of Exp-161: To a stirred solution of Exp-161b (330 mg, 0.42 mmol) and 1 (133 mg, 0.50 mmol) in DMF (5 mL) was added DIPEA (0.3 mL, 1.69 mmol). The reaction heated to 60 C. and stirred for 16 h. The reaction mass concentrated under reduced pressure and crude was purified by reverse phase combi flash column chromatography eluting with 10%-100% of ACN in H.sub.2O (0.1% Conc HCl) (compound isolated at 35% of ACN in H.sub.2O) to afford compound Exp-161 (200 mg, 47%) as a light yellow solid; .sup.1H NMR (400 MHz, CD.sub.3OD) 7.44-7.38 (m, 4H), 7.21-7.10 (m, 4H), 5.23-5.16 (m, 1H), 4.09-4.00 (m, 2H), 3.75-3.65 (m, 4H), 3.64-3.3.52 (m, 6H), 3.38-3.23 (m, 5H), 3.19-3.06 (m, 3H), 2.86 (t, J=7.42 Hz, 2H), 2.64 (t, J=6.8 Hz, 2H), 2.52 (t, J=7.34 Hz, 2H), 2.01-1.81 (m, 2H), 1.75-1.58 (m, 6H), 1.30-1.20 (m, 2H); ESI (m/z) [C.sub.43H.sub.64ClN.sub.13O.sub.12+H].sup.1 990; HPLC, AUC=99.5%; t.sub.R=6.12 min, Method P.

Example 162

Preparation of (S)-3,5-diamino-N(N-(4-(4-(3-((5-amino-1-(1H-tetrazol-5-yl)pentyl) (methyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-162)

##STR00522##

[0912] Preparation of Exp-162a: To a solution Int-51 (400 mg, 0.57 mmol) in IPA (10 mL) and EtOAc (10 mL), was added 20% Pd(OH).sub.2 on carbon 50% wet (200 mg, 50% weight substrate) under inert atmosphere. The resultant reaction mixture stirred under H.sub.2 (balloon) for 16 h. The reaction mass filtered through celite bed and filtrate concentrated under reduced pressure to afford Exp-162a (310 mg, 95%) as a light brown solid; ESI (m/z) [C.sub.31H.sub.45N.sub.7O.sub.3+H].sup.+ 564.

[0913] Preparation of Exp-162b: A stirred solution of Exp-162a (310 mg, 0.55 mmol) and compound 1 (235 mg, 0.60 mmol) in DMF (10 mL), was charged with DIPEA (0.27 mL, 1.65 mmol) at room temperature. The resultant reaction mixture was heated to 60 C. and stirred for 8 h. The reaction mixture concentrated under reduced pressure, water (50 mL) added, and the resulting suspension was stirred for 30 minutes. The solids were collected by filtration and washed with water (20 mL). Obtained solid azeotroped with CH.sub.3CN (25 mL) and dried under reduced pressure to afford Exp-162b (350 mg, 82%) as a brown solid; ESI (m/z) [C.sub.37H.sub.50ClN.sub.13O.sub.4+H].sup.+ 776.

[0914] Preparation of Exp-162: A stirred solution of Exp-162b (300 mg, 0.39 mmol) in MeOH (5.0 mL) was charged with 4M HCl in MeOH (5.0 mL) at room temperature then stirred for 6 h. The reaction mixture was concentrated under reduced pressure. The obtained crude material was purified by reverse phase combi-flash chromatography eluted with 10%-100% CH.sub.3CN in water (compound isolated at 35% of CH.sub.3CN) to afford Exp-162 (115 mg, 39%) as a light-yellow solid. .sup.1H NMR (400 MHz, CD.sub.3OD) 7.52-7.47 (m, 4H), 7.29-7.27 (m, 4H), 6.07 (dd, J=9.6, 6.0 Hz, 1H), 3.38-3.26 (m, 3H), 2.99 (t, J=7.6 Hz, 2H) 2.93 (t, J=7.2 Hz, 2H), 2.83 (s, 3H), 2.82-2.72 (m, 3H), 2.25-2.04 (m, 2H), 1.79-1.69 (m, 6H), 1.47-1.37 (m, 2H); ESI (m/z) [C.sub.32H.sub.42ClN.sub.13O.sub.2+H].sup.+ 676; HPLC, AUC=97.7%; t.sub.R=7.09 min, Method Q.

Example 163

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(3-(methyl((S)-5-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)-1-(1H-tetrazol-5-yl)pentyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl) butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-163)

##STR00523##

[0915] Preparation of Exp-163a: A stirred solution of Int-51 (900 mg, 1.29 mmol) in MeOH (5.0 mL) was added 4M HCl in MeOH (5.0 mL) slowly then stirred at room temperature for 3 h. The reaction mass was concentrated under reduced pressure, the residue was azeotroped with dichloromethane (25 mL) and dried under reduced pressure to afford Exp-163a (810 mg, 98%) as a yellow solid; ESI (m/z) [C.sub.34H.sub.43N.sub.7O.sub.3+H].sup.+ 598.

[0916] Preparation of Exp-163b: To a stirred solution of Exp-163a (800 mg, 1.26 mmol) in MeOH (15 mL), was added D-(+) glucose (249 mg, 1.38 mmol), AcOH (0.079 mL, 1.38 mmol) and NaBH.sub.3CN (86 mg, 1.38 mmol). The reaction mixture was warmed to 45 C. and stirred for 16 h. The reaction mass cool to room temperature and charged with D-(+) glucose (113 mg, 0.63 mmol) followed by NaBH.sub.3CN (39 mg, 0.63 mmol) then heated to 45 C. and stirred for another 16 h. The reaction mass concentrated under reduced pressure. The obtained crude compound treated with 4N HCl in CH.sub.3OH (2 mL) stirred for 15 min and concentrated under reduced pressure. Crude material was purified by reverse phase combi flash chromatography eluted with 10%-100% CH.sub.3CN in H.sub.2O (0.01% Conc. HCl) (compound isolated at 30% of CH.sub.3CN) to afford Exp-163b (300 mg, 31%) as a colorless gummy solid: ESI (m/z) [C.sub.40H.sub.55N.sub.7O.sub.8+H].sup.+ 762.

[0917] Preparation of Exp-163c: A stirred solution of Exp-163b (300 mg, 0.39 mmol) in IPA (10 mL) and water (10 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (100 mg, 50% weight substrate) under inert atmosphere. The resultant reaction mixture stirred under H.sub.2 (balloon) for 3 h. The reaction mass filtered through celite bed and filtrate concentrated under reduced pressure. The obtained crude was azeotroped with ACN (210 mL) and dried under reduced pressure to afford Exp-163c (210 mg, 85%) as a colorless gum; ESI (m/z) [C.sub.32H.sub.49N.sub.7O.sub.6+H].sup.+ 628.

[0918] Preparation of Exp-163: To a stirred solution of compound Exp-163c (210 mg, 0.33 mmol) and 1 (142 mg, 0.368 mmol) in DMF (5 mL) was added DIPEA (0.16 mL, 1.00 mmol). The reaction mixture was warmed to 60 C. and stirred for 16 h. The reaction mass was concentrated under reduced pressure then purified by reverse phase combi flash column chromatography eluted with 10%-100% of CH.sub.3CN in H.sub.2O (0.1% Conc. HCl) (compound isolated at 35% of CH.sub.3CN) to afford compound Exp-163 (95 mg, 31%) as a light yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.43-7.40 (m, 4H), 7.20-7.18 (m, 4H), 5.97 (dd, J=9.6, 6.0 Hz, 1H), 3.95-3.93 (m, 1H), 3.75-3.53 (m, 5H), 3.28-3.25 (m, 2H), 3.05-3.03 (m, 2H), 2.91-2.87 (m, 4H), 2.73-2.62 (m, 7H), 2.12-1.95 (m, 2H), 1.72-1.63 (m, 6H), 1.35-1.21 (m, 2H); ESI (m/z) [C.sub.38H.sub.54ClN.sub.13O.sub.7+H].sup.+ 840; HPLC, AUC=95.7%; t.sub.R=9.40 min, Method V.

Example 164

Preparation of 3,5-diamino-N(N-(4-(4-(3-(((S)-5-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxy hexyl)amino)-1-(1H-tetrazol-5-yl)pentyl)(methyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl) carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-164)

##STR00524##

[0919] Preparation of Exp-164a: To a solution of Exp-163a (0.60 g, 1.00 mmol) in MeOH (15.0 mL) was charged with D(+) glucose (0.54 g, 3.01 mmol) followed by AcOH (0.18 mL, 3.01 mmol) and NaBH.sub.3CN (0.19 g, 3.01 mmol). The resultant reaction mixture was stirred at 60 C. for 16 h. After 16 h, reaction mixture was cooled to ambient temperature and charged with D(+) glucose (0.36 g, 2.01 mmol) followed by NaBH.sub.3CN (0.13 g, 2.01 mmol) then stirred for another 16 h at 60 C. The reaction mixture was concentrated under reduced pressure and the obtained crude was treated with 4N HCl in CH.sub.3OH (5.0 mL), stirred for 15 minutes and concentrated. Crude material was purified by reverse phase combi-flash chromatography eluted with 10%-100% CH.sub.3CN in H.sub.2O (0.05% Conc. HCl) (compound isolated at 40% of CH.sub.3CN) to afford Exp-164a (0.65 g, 69%) as a light yellow solid: ESI (m/z) [C.sub.46H.sub.67N.sub.7O.sub.13+H].sup.+ 926.

[0920] Preparation of Exp-164b: To a solution of Exp-164a (0.65 g, 0.70 mmol) in IPA (10.0 mL) and water (10.0 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (0.20 g, 50% weight substrate) under inert atmosphere. The resultant reaction mixture stirred under H.sub.2 (balloon) for 3 h. The reaction mass filtered through celite bed and filtrate concentrated under reduced pressure to afford Exp-164b (0.50 g, 90%) as an colourless liquid; ESI (m/z) [C.sub.38H.sub.61N.sub.7O.sub.11+H].sup.+ 792.

[0921] Preparation of Exp-164: To a solution of compound Exp-164b (0.50 g, 0.63 mmol) and compound 1 (0.27 g, 0.69 mmol) in DMF (6.0 mL) was added DIPEA (0.12 mL, 0.94 mmol) at ambient temperature. The resultant reaction mixture was heated to 60 C. and stirred for 8 h. The reaction mixture concentrated under reduced pressure and the crude was purified by reverse phase combi flash chromatography eluting with 10%-100% CH.sub.3CN in H.sub.2O (0.01% Conc. HCl) (compound isolated at 30% of CH.sub.3CN) to afford Exp-164 (0.11 g, 16%) as a light-yellow solid. .sup.1H NMR (400 MHz, CD.sub.3OD) 7.45-7.37 (m, 4H), 7.22-7.18 (m, 4H), 6.03-5.95 (m, 1H), 4.09-4.02 (m, 2H), 3.86-3.78 (i, 1H), 3.76-3.66 (m, 4H), 3.63-3.53 (m, 6H), 3.40-3.30 (m, 2H), 3.29-3.24 (m, 7H), 2.93-2.87 (i, 2H), 2.75-2.61 (i, 7H), 2.15-1.98 (m, 2H), 1.79-1.61 (i, 6H), 1.36-1.22 (m, 2H); ESI (m/z) [CH.sub.66ClN.sub.13O.sub.2+H].sup.+ 1004; HPLC, AUC=99.3%; t.sub.R=6.16 min, Method P.

Examples 165-170

[0922] Preparation of Exp-165-170: In an analogous sequence of procedures used for Examples 159-164, Examples 165-170 were synthesized substituting D-N(CH.sub.3)-lysine for L-N(CH.sub.3)-lysine:

##STR00525##

TABLE-US-00042 Exp # R.sup.8 R R.sup.1 Spectra 165 H H H .sup.1H NMR (400 MHz, CD.sub.3OD) 7.40 (d, J = 8.4 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 7.18-7.13 (m, 4H), 5.20 (t, J = 7.2 Hz, 1H), 4.78-4.72 (m, 2H), 2.87-2.82 (m, 2H), 2.75 (t, J = 7.6 Hz, 2H), 2.62 (t, J = 7.2 Hz, 2H), 2.46 (t, J = 7.6 Hz, 2H), 1.77-1.52 (m, 8H), 1.22-1.16 (m, 2H) ESI (m/z) [C.sub.31H.sub.40ClN.sub.13O.sub.2 + H].sup.+ 662; HPLC; AUC = 97.7%; t.sub.R = 7.06 min; Method R. 166 H Sug H .sup.1H NMR (400 MHz, CD.sub.3OD) 7.42 (d, J = 8.4 Hz, 2H), 7.38 (d, J = 8.0 Hz, 2H), 7.20 (d, J = 8.4 Hz, 2H), 7.14 (d, J = 8.0 Hz, 2H), 5.16- 5.15 (m, 1H), 3.94-3.92 (m, 1H), 3.74-3.52 (m, 5H), 3.28-3.25 (m, 2H), 3.01-2.95 (m, 2H), 2.88-2.80 (m, 4H), 2.64 (t, J = 7.2 Hz, 2H), 2.53-2.50 (m, 2H), 1.98-1.80 (m, 2H), 1.73-1.60 (m, 6H), 1.30-1.19 (m, 2H); ESI (m/z) [C.sub.37H.sub.52ClN.sub.13O.sub.7 + H].sup.+ 826; HPLC, AUC = 95.7%; t.sub.R = 6.18 min, Method P. 167 H Sug Sug .sup.1H NMR (400 MHz, CD.sub.3OD) 7.43 (d, J = 8.4 Hz, 2H), 7.39 (d, J = 8.4 Hz, 2H), 7.21-7.14 (m, 4H), 5.21-5.18 (m, 1H), 4.06-4.02 (m, 2H), 3.74-3.54 (m, 10H), 3.35-3.23 (m, 7H), 3.12-3.02 (m, 1H), 2.87 (t, J = 7.2 Hz, 2H), 2.64 (t, J = 7.2 Hz, 2H), 2.54-2.50 (m, 2H), 2.00-1.82 (m, 2H), 1.72-1.62 (m, 6H), 1.30-1.19 (m, 2H); ESI (m/z) [C.sub.43H.sub.64ClN.sub.13O.sub.12 + H].sup.+ 990; HPLC, AUC = 93.2%; t.sub.R = 9.69 min, Method U. 168 CH.sub.3 H H .sup.1H NMR (400 MHz, CD.sub.3OD) 7.42-7.37 (m, 4H), 7.23-7.18 (m, 4H), 5.97 (dd, J = 6.0, 9.6 Hz, 1H), 3.28-3.25 (m, 2H), 2.91-2.81 (m, 4H), 2.73 (s, 3H), 2.70-2.62 (m, 4H), 2.11-1.96 (m, 2H), 1.73-1.61 (m, 6H), 1.37-1.19 (m, 2H); ESI (m/z) [C.sub.32H.sub.42ClN.sub.13O.sub.2 + H].sup.+ 676; HPLC, AUC = 99.5%; t.sub.R = 7.08 min, Method R. 169 CH.sub.3 Sug H .sup.1H NMR (400 MHz, CD.sub.3OD) 7.43-7.38 (m, 4H), 7.24-7.19 (m, 4H), 5.99-5.96 (m, 1H), 3.96-3.92 (m, 1H), 3.74-3.67 (m, 2H), 3.62-3.55 (m, 3 H), 3.28-3.25 (m, 2 H), 3.05-3.04 (m, 2 H), 2.91-2.88 (m, 4 H), 2.73-2.62 (m, 7H), 2.09-1.98 (m, 2H), 1.72-62 (m, 6H), 1.32-1.28 (m, 2H); ESI (m/z) [C.sub.38H.sub.54ClN.sub.13O.sub.7 + H].sup.+ 840; HPLC AUC 91.6% (R.sub.t = 7.79 min); HPLC, AUC = 91.7%; t.sub.R = 7.79 min, Method W. 170 CH.sub.3 Sug Sug .sup.1H NMR (400 MHz, CD.sub.3OD) 7.45-7.37 (m, 4H), 7.23-7.17 (m, 4H), 6.04-5.95 (m, 1H), 4.10-4.03 (m, 2H), 3.76-3.66 (m, 4H), 3.64-3.53 (m, 6H), 3.40-3.33 (m, 1H), 3.30-3.23 (m, 7H), 2.93-2.87 (m, 2H), 2.75-2.60 (m, 7H), 2.15-1.98 (m, 2H), 1.79- 1.61 (m, 6H), 1.36-1.22 (m, 2H); ESI (m/z) [C.sub.46H.sub.66ClN.sub.13O.sub.12 + H].sup.+ 1005; HPLC, AUC = 97.9%; t.sub.R = 7.63 min, Method W.

Example 171

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(3-oxo-3-((2-(((2R,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl) amino)propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-171)

##STR00526## ##STR00527##

[0923] Preparation of Exp-171a: To a stirred solution of Int-34 (2.80 g, 5.49 mmol) in MeOH (100 mL) was added D(+)-glucose (1.48 g, 5.4 mmol) followed by pic-BH.sub.3 (872 mg, 8.23 mmol). Then resultant reaction mixture was stirred at 60 C. for 24 h. The reaction mixture was cooled to room temperature, charged with D(+)-glucose (494 mg, 2.74 mmol) and pic-BH.sub.3 (582 mg, 5.49 mmol). The resultant reaction heated to 60 C. and stirred for 16 h. The reaction mass was concentrated under reduced pressure and to the obtained crude was added 50% CH.sub.3CN in H.sub.2O (20 mL). The resulting suspension was filtered the solids were dried to yield compound Exp-171a (2.10 g, 60%) as a brown solid: ESI (m/z) [C.sub.35H.sub.47N.sub.3O.sub.8+H].sup.+ 638.

[0924] Preparation of Exp-171b: To a solution of Exp-171a (2.10 g, 3.29 mmol) and L()Mannose (888 mg, 4.93 mmol) in MeOH (100 mL) was charged AcOH (0.29 mL, 4.93) followed by NaBH.sub.3CN (306 mg, 4.93 mmol). The resultant reaction mixture was stirred at 60 C. for 24 h. The reaction mixture was cooled to room temperature, charged with L()Mannose (590 mg, 3.29 mmol) and NaBH.sub.3CN (203 mg, 3.29 mmol). The resultant reaction heated to 60 C. and stirred for 24 h. The reaction mixture was charged with L()Mannose (590 mg, 3.29 mmol), NaBH.sub.3CN (203 mg, 3.29 mmol) and AcOH (0.19 mL, 3.29) once a day for 8 days (with stirring at 60 C. between charges). The reaction mass concentrated under reduced pressure, the crude treated with 4M HCl in MeOH (1.0 mL) in MeOH (10 mL), concentrated and azeotroped with MeOH (10 mL). The obtained crude was purified by C-18 reverse phase combi flash purification eluting with 10-100% CH.sub.3CN in H.sub.2O (0.05% HCl) (compound isolated at 40% of CH.sub.3CN to afford Exp-171b (450 mg, 25%) as a colourless gum: ESI (m/z) [C.sub.41H.sub.59N.sub.3O.sub.13+H].sup.+ 802.

[0925] Preparation of Exp-171c: To a solution Exp-171b (1.40 g, 1.74 mmol) in IPA (20 mL) and H.sub.2O (20 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (300 mg, 50% weight substrate) followed by AcOH (0.3 mL) under inert atmosphere. The resultant reaction mixture stirred under H.sub.2 (balloon) for 4 h. The reaction mass was filtered through celite bed and filtrate was concentrated under reduced pressure to afford Exp-171c (800 mg, 63%) as a colourless gum; ESI (m/z) [C.sub.33H.sub.53N.sub.3O.sub.11+H].sup.+ 668.

[0926] Preparation of Exp-171: To a solution of compound Exp-171c (0.80 g, 1.1 mmol) and compound 1 (0.469 g, 1.21 mmol) in DMF (15.0 mL) was charged with DIPEA (0.72 mL, 4.4 mmol) at ambient temperature. The resultant reaction mixture was warmed to 60 C. and stirred for 16 h. The reaction mixture was concentrated under reduced pressure and the crude was purified by reverse phase combi-flash chromatography eluting with 10%-100% CH.sub.3CN in H.sub.2O (0.01% Conc. HCl) (compound isolated at 35% of CH.sub.3CN) to afford Exp-171 (560 mg, 57%) as a light yellow solid; .sup.1H NMR (400 MHz, CD.sub.3OD) 7.43-7.40 (m, 4H), 7.19-7.17 (m, 4H), 4.19-4.10 (m, 1H), 4.02-3.98 (m, 1H), 3.78-3.69 (m, 4H), 3.61-3.38 (m, 11H), 3.35-3.22 (m, 5H), 2.86 (t, J=8.0 Hz, 2H), 2.63 (t, J=7.2 Hz, 2H), 2.48 (t, J=8.0 Hz, 2H), 1.79-1.62 (m, 4H); ESI (m/z) [C.sub.39H.sub.58ClN.sub.9O.sub.12+H].sup.+ 880; HPLC AUC>99% (R.sub.t=7.44 min); HPLC, AUC=99.7%; t.sub.R=7.44 min, Method T.

Example 172

Preparation of (S)-3,5-diamino-N(N-(4-(4-(3-(2-(4-aminobutyl)-3-oxopiperazin-1-yl)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-172)

##STR00528##

[0927] Preparation of Exp-172a: To compound Int-54 (1.26 g, 1.542 mmol) in 250 mL rbf was added IPA/MeOH/H.sub.2O (60 mL/20 mL/10 mL) under nitrogen and the suspension was stirred at 50 C. for 2 h resulting in a clear solution. Pd(OH).sub.2/C (20% on carbon, 200 mg, 50% wet) was added, and the nitrogen balloon was switched to hydrogen balloon. The suspension was stirred at 50 C. for 4 h. After filtration, the solvent was removed, to afford Exp-172a (972 mg, 85%) as colorless oil: ESI (m/z) [C.sub.38H.sub.58N.sub.4O.sub.7+H].sup.+ 683.

[0928] Preparation of Exp-172b: To a solution of Exp-172a (972 mg, 1.31 mmol) in DMF (20 mL) was added compound 1 (508 mg, 1.31 mmol) and DIPEA (845 mg, 6.54 mmol). The resulting solution was warmed to 55 C. and stirred for 16 hr. After the solvent was removed, the residue was purified by reverse phase column (10% to 90% of CH.sub.3CN in water, product came out from 60% to 90%), to afford Exp-172b (250 mg, 21%) as a yellow solid: ESI (m/z) [C.sub.44H-63ClN.sub.10O.sub.8+H].sup.+ 895.

[0929] Preparation of Exp-172c: To a solution of Exp-172b (150 mg, 0.168 mmol) in MeOH (2.0 mL) was added 3 N HCl in MeOH (2.0 mL). The resulting solution was stirred at rt for 3 hr. The solvent was removed, to afford Exp-172c (135 mg, 100%) as a yellow solid: ESI (m/z) [C.sub.34H.sub.47ClN.sub.10O.sub.4+H].sup.+ 695.

[0930] Preparation of Exp-172: To a solution of Exp-172c (139 mg, 0.056 mmol) in CH.sub.3CN/water (6.0 mL/2.0 mL) was added DIPEA (151 mg, 1.16 mmol). The resulting solution was stirred at rt for 1 hr. After the solvent was removed, the residue was suspended in CH.sub.3CN/water (1.0 mL/2.0 mL) and the pH was adjusted to 2 with 1 N HCl. A clear yellow solution was observed. The solution was purified by reverse phase column (5% to 70% of CH.sub.3CN in water, product came out at 20%), to afford Exp-172 (56 mg, 39%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.50 (m, 4H), 7.31-7.27 (m, 4H), 4.93-4.90 (m, 1H), 4.00-3.90 (m, 1H), 3.44-3.15 (m, 5H), 2.98-2.71 (m, 8H), 1.91-1.66 (m, 8H), 1.45-1.41 (m, 2H); ESI (m/z) [C.sub.33H.sub.43ClN.sub.10O.sub.3+H].sup.+ 663; HPLC purity 94.2%; HPLC, AUC=94.2%; t.sub.R=9.38 min, Method A.

Example 173

Preparation of N.SUP.2.-(2-aminoethyl)-N2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-L-lysine (Exp-173)

##STR00529##

[0931] Preparation of Exp-173a: To a solution of Exp-172b (50.0 mg, 0.056 mmol) in THF/MeOH/water (0.9 mL/0.9 mL/0.3 mL) was added NaOH (22.3 mg, 0.558 mmol). The resulting solution was stirred at rt for 3 hr. The solvent was removed, to afford Exp-173a (72 mg, crude) as a yellow solid: ESI (m/z) [C.sub.43H.sub.61ClN.sub.10O.sub.8+H].sup.+ 881.

[0932] Preparation of Exp-173: Exp-173a (72 mg, crude) was dissolved in 4 N HCl in dioxane (0.5 mL) and 4N HCl (0.5 mL). The resulting solution was stirred at rt for 3 hr. After the solvent was removed, the residue was re-suspended in CH.sub.3CN/water (1.0 mL/2.0 mL) and purified by reverse phase column (5% to 60% of CH.sub.3CN in water, product came out at 20%), to afford Exp-173 (31 mg, 69%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.49 (m, 4H), 7.33-7.27 (m, 4H), 4.45 (br s, 0.6H), 4.20 (br s, 0.3H), 3.95-3.50 (m, 2H), 3.40-3.10 (m, 5H), 2.99-2.71 (m, 8H), 2.13-2.10 (m, 1H), 1.81-1.60 (m, 6H), 1.45-1.37 (m, 2H); ESI (m/z) [C.sub.33H.sub.45ClN.sub.10O.sub.4+H].sup.+ 681; HPLC purity 95.5%; HPLC, AUC=95.5%; t.sub.R=8.72 min, Method A.

Example 174

Preparation of (S)-methyl 6-amino-2-(N-(2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxy hexyl)amino)ethyl)-3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)hexanoate (Exp-174)

##STR00530## ##STR00531##

[0933] Preparation of Exp-174a: To a solution of Int-55 (150 mg, 0.153 mmol) in CH.sub.2Cl.sub.2 (3.0 mL) was added piperidine (0.5 mL) at rt. The reaction mixture was stirred at rt for 16 h. After the solvent removed, the residue was precipitated from MTBE/hexanes (5.0 mL/5.0 mL), the solvent was decanted and the precipitate was dried in high vacuum, to afford Exp-174a (102 mg, 88%) as yellow oil: ESI (m/z) [C.sub.44H.sub.62N.sub.4O.sub.7+H].sup.+ 759.

[0934] Preparation of Exp-174b: To a solution of Exp-174a (100 mg, 0.132 mmol) and glucose (71.2 mg, 0.395 mmol) in MeOH (2.0 mL) was added NaCNBH.sub.3 (24.8 mg, 0.395 mmol) and AcOH (23.7 mg, 0.395 mmol) at rt. The reaction mixture was stirred at 60 C. for 16 h. The solvent was removed and water (2.0 mL) was added resulting in precipitation. The water phase was decanted, and the residue was azeotroped with MeOH, to afford 108 mg of crude product, which was further purified by reverse phase column (10% to 90% of CH.sub.3CN in water, product came out from 60% to 80%), to afford Exp-174b (46.0 mg, 32%) as colorless syrup: ESI (m/z) [C.sub.56H.sub.86N.sub.4O.sub.17+H].sup.+ 1087.

[0935] Preparation of Exp-174c: To a solution of compound Exp-174b (190 mg, 0.175 mmol) in IPA/MeOH/H.sub.2O (1.0 mL/1.0 mL/1.0 mL) was added Pd(OH).sub.2/C (30 mg) at rt. The reaction mixture was stirred under hydrogen (balloon) at 45 C. for 6 h. Almost no reaction was observed. After filtration, fresh Pd(OH).sub.2/C was added and the reaction mixture was stirred under hydrogen (balloon) at 45 C. for 8 h. After filtration, fresh Pd(OH).sub.2/C and AcOH (6.0 equiv) were added and the reaction mixture was stirred under hydrogen (balloon) at 45 C. for 8 h. After filtration, fresh Pd(OH).sub.2/C was added and the reaction mixture was stirred under hydrogen (balloon) at 50 C. for 4 h. After filtration, the solvent was removed, to afford compound Exp-174c (168 mg, 90%) as colorless syrup: ESI (m/z) [C.sub.48H.sub.80N.sub.4O.sub.15+H].sup.+ 953.

[0936] Preparation of Exp-174d: To a solution of compound Exp-174c (168 mg, 0.157 mmol) in DMF (3.0 mL) was added compound 1 (60.8 mg, 0.157 mmol) and DIPEA (60.7 mg, 0.470 mmol). The resulting solution was stirred at 55 C. for 16 hr. After the solvent was removed, the residue was precipitated from IPA (5.0 mL). The IPA phase was decanted and the solids were dried to afford compound Exp-174d (164 mg, 90%) as a yellow solid: ESI (m/z) [C.sub.54H.sub.85ClN.sub.10O.sub.16+H].sup.+ 1165.

[0937] Preparation of Exp-174: To a solution of Exp-174d (234 mg, 0.201 mmol) in MeOH (3.0 mL) was added 3 N HCl in MeOH (3.0 mL). The resulting solution was stirred at 40 C. for 16 hr. After the solvent was removed and azeotroped with 1 N HCl in MeOH (4.0 mL2) to remove boron, the residue was purified by reverse phase column (10% to 70% of CH.sub.3CN in water, product came out at 20% to 30%), to afford Exp-174 (15 mg, 6%) as a yellow solid. Additional 186 mg of acid/ester mixture was obtained: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.54-7.51 (m, 4H), 7.33-7.28 (m, 4H), 4.72-4.70 (m, 1H), 4.22-4.20 (m, 2H), 3.86-3.40 (m, 23H), 2.99-2.71 (m, 8H), 2.15-2.05 (m, 1H), 1.79-1.68 (m, 7H), 1.46-1.38 (m, 2H); ESI (m/z) [C.sub.46H.sub.71ClN.sub.10O.sub.14+H].sup.+ 1023; HPLC, AUC=95.4%; t.sub.R=8.68 min, Method A.

Example 175

Preparation of (S)-6-amino-2-(N-(2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) amino)ethyl)-3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)hexanoic acid (Exp-175)

##STR00532##

[0938] Preparation of Exp-175: To a solution of Exp-174 (186 mg, 0.164 mmol, mixture from previous experiment) in MeOH (2.0 mL) and water (2.0 mL) was added NaOH (65.7 mg, 1.64 mmol). The resulting solution was stirred at rt for 3 hr. The pH was adjusted to 2, and the solvent was removed. The residue was purified by reverse phase column (5% to 70% of CH.sub.3CN in water, product came out at 30%), to afford Exp-175 (142 mg, 77%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.48 (m, 4H), 7.30-7.27 (m, 4H), 4.22-4.20 (m, 3H), 3.90-3.60 (m, 14H), 3.50-3.32 (m, 6H), 2.98-2.90 (m, 4H), 2.75-2.71 (m, 4H), 2.10-2.00 (m, 1H), 1.79-1.66 (m, 7H), 1.45-1.30 (m, 2H); ESI (m/z) [C.sub.45H.sub.69ClN.sub.10O.sub.14+H].sup.+ 1009; HPLC, AUC=98.5%; t.sub.R=8.63 min, Method A.

Example 176

Preparation of 3,5-diamino-N(N-(4-(4-(3-(((S)-5-amino-1-(1H-tetrazol-5-yl)pentyl)(2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-176)

##STR00533## ##STR00534##

[0939] Preparation of Exp-176a: A solution of Int-56 (400 mg, 0.417 mmol) in CH.sub.2Cl.sub.2 (10 mL) was charged with piperidine (0.5 mL) then stirred at room temperature for 3 h. The reaction mass was concentrated under reduced pressure and the obtained crude solid was washed with 30% MTBE in hexanes (310 mL). The solid was dried under reduced pressure to afford compound Exp-176a (280 mg, 59%) as a light yellow gum; ESI (m/z) [C.sub.40H.sub.54N.sub.8O.sub.5+H].sup.+ 727

[0940] Preparation of Exp-176b: To a solution of Exp-176a (280 mg, 0.385 mmol) and D(+)-glucose (277 mg, 1.54 mmol) in MeOH (15 mL) was charged AcOH (92.4 mg, 1.54 mmol) followed by NaBH.sub.3CN (95.5 mg, 1.54 mmol). The resultant reaction mixture was stirred at 60 C. for 16 h. The reaction mixture was concentrated under reduced pressure and crude was purified by C-18 reverse phase combi flash purification eluted with 10-100% CH.sub.3CN in H.sub.2O (0.05% HCOOH) (compound isolated at 45% of CH.sub.3CN) to afford Exp-176b (200 mg, 49%) as a colourless gum: ESI (m/z) [C.sub.52H.sub.78N.sub.8O.sub.15+H].sup.+ 1055.

[0941] Preparation of Exp-176c: To a solution of Exp-176b (200 mg, 0.189 mmol) in IPA (5 mL) and H.sub.2O (5 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (50 mg, 50% weight substrate) under inert atmosphere. The resultant reaction mixture stirred under H.sub.2 (balloon) for 3 h. The reaction mass was filtered through a celite bed and the filtrate was concentrated under reduced pressure. The obtained crude was azeotroped with CH.sub.3CN (10 mL) and dried under reduced pressure to afford Exp-176c (130 mg, 75%) as a colorless gum; ESI (m/z) [C.sub.44H.sub.72N.sub.8O.sub.13+H].sup.+ 921.

[0942] Preparation of Exp-176d: To a solution of Exp-176c (130 mg, 0.141 mmol) and compound 1 (54.8 mg, 0.141 mmol) in DMF (5 mL) was added DIPEA (0.069 mL, 0.423 mmol). The reaction was heated to 60 C. and stirred for 16 h. The reaction mass was concentrated under reduced pressure, the obtained crude was washed with MTBE (10 mL) and dried under reduced pressure to afford Exp-176d (200 mg (rude)) as a yellow gum: ESI (m/z) [C.sub.50H.sub.77ClN.sub.14O.sub.14+H].sup.+ 1133.

[0943] Preparation of Exp-176: To a stirred solution of compound Exp-176d (200 mg (crude)) in MeOH (5 mL) was charged with 4M HCl in MeOH (2.0 mL). The reaction mixture was stirred at room temperature for 8 h. The reaction mixture was concentrated under reduced pressure. The obtained crude was purified by C-18 reverse phase combi-flash chromatography eluting with 10%-100% CH.sub.3CN in water (0.5% HCl) (compound isolated at 35% of CH.sub.3CN) to afford compound Exp-176 (95 mg, 55% over two steps) as an off-white solid: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.43-7.40 (m, 4H), 7.24-7.18 (m, 4H), 5.42 (t, J=7.2 Hz, 1H), 4.12-4.03 (m, 2H), 3.72-3.52 (m, 12H), 3.40-3.32 (m, 5H), 3.25-3.18 (m, 3H), 2.94-2.90 (m, 6H), 2.65-2.61 (m, 2H), 2.22-2.18 (m, 1H), 2.01-1.95 (m, 1H), 2.72-2.65 (m, 6H), 1.38-1.30 (m, 2H); ESI (m/z) [C.sub.45H.sub.69ClN.sub.14O.sub.12+H].sup.+ 1033; HPLC AUC 98.2% (R.sub.t=7.15 min); HPLC, AUC=98.3%; t.sub.R=7.15 min, Method W.

Example 177-178

[0944] Preparation of Exp-177-178: In an analogous sequence of procedures used for Examples 174-175, Examples 177-178 were synthesized substituting D-lysine for L-lysine at the beginning of the reaction scheme:

##STR00535##

TABLE-US-00043 Exp # R.sup.9 Spectra 177 CH.sub.3 .sup.1H NMR (400 MHz, CD.sub.3OD) 7.45-7.41 (m, 4H), 7.24-7.18 (m, 4H), 4.65- 4.61 (m, 1H), 4.15-4.02 (m, 3H), 3.78-3.75 (m, 2H), 3.64 (s, 3H), 3.62-3.52 (m, 10H), 3.48-3.38 (m, 5H), 3.28-3.22 (m, 2H), 2.88-2.80 (m, 4H), 2.79- 2.62 (m, 4H), 2.01-1.98 (m, 1H), 1.89-1.55 (m, 7H), 1.40-1.22 (m, 2H); ESI (m/z) [C.sub.46H.sub.71ClN.sub.10O.sub.14 + H].sup.+ 1023; HPLC AUC 95.8 % (R.sub.t = 7.19 min); HPLC, AUC = 95.7%; t.sub.R = 7.19 min, Method W. 178 H .sup.1H NMR (400 MHz, CD.sub.3OD) 7.41 (t, J = 8.4 Hz, 4H), 7.20-7.18 (m, 4H), 4.18-4.02 (m, 3H), 3.74-3.67 (m, 5H), 3.62-3.52 (m, 7H), 3.49-3.31 (m, 5H), 3.29-3.20 (m, 3H), 2.89-2.80 (m, 4H), 2.72-2.63 (m, 4H), 2.02-1.93 (m, 1H), 1.70-1.50 (m, 7H), 1.32-1.22 (m, 2H); ESI (m/z) [C.sub.45H.sub.69ClN.sub.10O.sub.14 + H].sup.+ 1009; HPLC AUC 95.8 % (R.sub.t = 7.15 min); HPLC, AUC = 95.8%; t.sub.R = 7.15 min, Method W.

Example 179

Preparation of methyl N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N.SUP.2.-(3-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)propyl)-L-lysinate (Exp-179)

##STR00536## ##STR00537##

[0945] Preparation of Exp-179a: A solution of Int-56 (2.0 g, 2.09 mmol) in CH.sub.2Cl.sub.2 (25 mL) was charged with piperidine (8.0 mL) then stirred at room temperature for 2 h. The reaction mixture was concentrated at 25 C. under reduced pressure and the obtained crude solid washed with 1:1 MTBE and hexanes (320 mL). The resulting solid was dried under reduced pressure to afford Exp-179a (1.38 g, 85%) as an off white semi solid; ESI (m/z) [C.sub.42H.sub.58N.sub.4O.sub.7+H].sup.+ 731

[0946] Preparation of Exp-179b: To a solution of compound Exp-179a (1.3 g, 1.78 mmol) and D (+) glucose (320 mg, 1.78 mmol) in MeOH (30 mL) was charged AcOH (181 mg, 1.783.69) followed by NaBH.sub.3CN (111 mg, 1.78 mmol). The resultant reaction mixture was stirred at 45 C. for 24 h. The reaction mixture was again charged D (+) glucose (160 mg, 0.89 mmol) and NaBH.sub.3CN (56 mg, 0.859 mmol) at room temperature and heated to 45 C. and stirred for 36 h. The reaction mixture was concentrated under reduced pressure and the obtained crude material was purified by reverse phase combi flash purification eluting with 10-100% CH.sub.3CN in H.sub.2O (0.05% HCOOH) (compound isolated at 45% of CH.sub.3CN) to afford compound Exp-179b (550 mg, 37%) as a colourless gum: ESI (m/z) [C.sub.48H.sub.70N.sub.4O.sub.12.sup.+ H].sup.+ 895.

[0947] Preparation of Exp-179c: To a solution of compound Exp-179b (550 mg, 0.614 mmol) in a mixture of IPA (15 mL), EtOAc (10 mL) and H.sub.2O (5 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (200 mg, 50% weight substrate). The resultant reaction mixture was stirred under H.sub.2 (balloon) for 7 h. The reaction mixture was filtered through a celite bed and the filtrate was concentrated under reduced pressure. The obtained crude was azeotroped with toluene (220 mL) and dried under reduced pressure to afford compound Exp-179c (420 mg, crude) as a colourless gum; ESI (m/z) [C.sub.40H.sub.64N.sub.4O.sub.10+H].sup.+ 761.

[0948] Preparation of Exp-179d: To a solution of Exp-179c (420 mg, 0.469 mmol) and compound 1 (200 mg, 0.516 mmol) in DMF (10 mL), was added DIPEA (0.33 mL, 1.876 mmol). The reaction mixture was heated to 60 C. and stirred for 16 h. The reaction mixture was concentrated under reduced pressure and the obtained crude was purified by reverse phase combi flash purification eluting with 10-100% CH.sub.3CN in H.sub.2O (0.05% HCOOH) (compound isolated at 30% of CH.sub.3CN) to afford Exp-179d (380 mg, 70%) as a pale yellow solid: ESI (m/z) [C.sub.46H.sub.69ClN.sub.10O.sub.11+H].sup.+ 973.

[0949] Preparation of Exp-179: A stirred solution of Exp-179d (350 mg, 0.395 mmol) in MeOH (10 mL) was charged with 4M HCl in MeOH (5.0 mL) then stirred at room temperature for 6 h. The reaction mixture was concentrated under reduced pressure and the obtained crude was purified by reverse phase combi-flash chromatography eluted with 10%-100% CH.sub.3CN in water (0.05% HCl) (compound isolated at 36% of CH.sub.3CN) to afford 100 mg of compound Exp-179 with HPLC-95.2% and 160 mg of a mixture of Exp-179 and Exp-180 (acid). Further, to a stirred solution of the mixture of acid and ester in methanol (1.0 mL) was added 4M HCl in MeOH (4 mL) and stirred at 45 C. for 6 h. The reaction mixture was concentrated under reduced pressure. The obtained crude was washed with MTBE (10 mL) and dried to afford compound Exp-179 (75 mg) as an light yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.46-7.40 (m, 4H), 7.25-7.16 (m, 4H), 4.61-4.52 (m, 0.2H), 4.08-4.00 (m, 0.6H), 4.00-3.93 (m, 1H), 3.78-3.72 (m, 1H), 3.71-3.62 (m, 2H), 3.62-3.51 (m, 5H), 3.31-3.24 (m, 4H), 3.13-2.98 (m, 3H), 2.93-2.78 (m, 5H), 2.74-2.68 (m, 1H), 2.67-2.58 (m, 3H), 2.08-1.90 (m, 2H), 1.88-1.52 (m, 8H), 1.43-1.23 (m, 2H); ESI (m/z) [C.sub.41H.sub.61ClN.sub.10O.sub.9+H].sup.+ 873; HPLC AUC 92.6% (R.sub.t=7.40 min); HPLC, AUC=92.6%; t.sub.R=7.40 min, Method W.

Example 180

Preparation of N.SUP.2.-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanoyl)-N2-(3-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)propyl)-L-lysine (Exp-180)

##STR00538##

[0950] Preparation of Exp-180: A stirred solution of compound Exp-179 (160 mg, 0.244 mmol) in water (5 mL) was charged with NaOH (73.2 mg, 1.831 mmol) then stirred at room temperature for 1 h. The reaction mixture acidified to pH=2 with 2N HCl solution and concentrated under reduced pressure. The obtained crude was purified by reverse phase combi-flash chromatography eluting with 10%-100% CH.sub.3CN in water (compound isolated at 20% of CH.sub.3CN) to afford compound Exp-180 (75 mg, 47%) as an off-white solid: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.45-7.38 (m, 4H), 7.25-7.16 (m, 4H), 4.31-4.23 (m, 0.5H), 4.15-4.09 (m, 0.3H), 3.99-3.92 (m, 1H), 3.76-3.71 (m, 1H), 3.70-3.63 (m, 1H), 3.62-3.51 (m, 5H), 3.29-3.23 (m, 2H), 3.11-2.77 (m, 8H), 2.75-2.57 (m, 4H), 2.07-1.82 (m, 3H), 1.74 (m, 7H), 1.42-1.20 (m, 2H); ESI (m/z) [C.sub.40H.sub.59ClN.sub.10O.sub.9+H].sup.+ 859; HPLC AUC 98.5% (R.sub.t=7.17 min); HPLC, AUC=98.5%; t.sub.R=7.17 min, Method W.

Example 181

Preparation of (S)-methyl 6-amino-2-(N-(3-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxy hexyl)amino)propyl)-3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)hexanoate (Exp-181)

##STR00539##

[0951] Preparation of Exp-181a: To a solution of Exp-179a (50.0 mg, 0.068 mmol) and glucose (37.0 mg, 0.205 mmol) in MeOH (2.0 mL) was added NaCNBH.sub.3 (12.9 mg, 0.205 mmol) and AcOH (12.3 mg, 0.205 mmol) at rt. The reaction mixture was stirred at 60 C. for 16 h. After the solvent was removed, the residue was purified by reverse phase column (10% to 90% of CH.sub.3CN in water, product came out at 50% to 90%), to afford compound Exp-181a (37.0 mg, 51%) as colorless syrup: ESI (m/z) [C.sub.54H.sub.82N.sub.4O.sub.17+H].sup.+ 1059.

[0952] Preparation of Exp-181: In steps analogous to those used to convert Exp-179c, to Exp-179, Exp-181a was converted to Exp-181: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.53-7.51 (m, 4H), 7.32-7.28 (m, 4H), 4.19-4.13 (m, 3H), 3.84-3.64 (m, 14H), 3.44-3.40 (m, 9H), 2.99-2.71 (m, 8H), 2.16-2.09 (m, 3H), 1.82-1.64 (m, 7H), 1.44-1.38 (m, 2H); ESI (m/z) [C.sub.47H.sub.73ClN.sub.10O.sub.14+H].sup.+ 1037; HPLC, AUC=100%; t.sub.R=9.03 min, Method A.

Example 182

Preparation of (S)-6-amino-2-(N-(3-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) amino)propyl)-3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)hexanoic acid (Exp-182)

##STR00540##

[0953] Preparation of Exp-182: In a procedure analogous to the synthesis of Exp-180, Exp-182 was prepared using similar conditions and reagents: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.52-7.49 (m, 4H), 7.31-7.27 (m, 4H), 4.20-4.17 (m, 3H), 3.83-3.64 (m, 11H), 3.44-3.30 (m, 9H), 2.98-2.90 (m, 5H), 2.73 (t, J=7.0 Hz, 3H), 2.08-2.02 (m, 3H), 1.79-1.65 (m, 7H), 1.40-1.39 (m, 2H); ESI (m/z) [C.sub.46H.sub.71ClN.sub.10O.sub.14+H].sup.+ 1023; HPLC purity 99.4%; HPLC, AUC=99.5%; t.sub.R=8.68 min, Method A.

Example 183

Preparation of 3,5-diamino-N(N-(4-(4-(3-(((S)-5-amino-1-(1H-tetrazol-5-yl)pentyl)(3-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)propyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-183)

##STR00541## ##STR00542##

[0954] Preparation of Exp-183a: A solution of Int-58 (800 mg, 0.83 mmol) in CH.sub.2Cl.sub.2 (20 mL) was charged with piperidine (1.0 mL) then stirred at room temperature for 3 h. The reaction mass was concentrated under reduced pressure and the obtained crude solid washed with MTBE (310 mL). The solid was dried under reduced pressure to afford Exp-183a (500 mg, 81%) as a light brown solid; ESI (m/z) [C.sub.41H.sub.56N.sub.8O.sub.5+H].sup.+ 741

[0955] Preparation of Exp-183b: To a solution of Exp-183a (900 mg, 1.21 mmol) and D(+)-glucose (218 mg, 1.21 mmol) in MeOH (20 mL) was charged with AcOH (87 mg, 1.45 mmol) followed by NaBH.sub.3CN (90 mg, 1.45 mmol). The resultant reaction mixture was stirred at 45 C. for 16 h. The reaction mixture was cooled to room temperature and charged with D(+)-glucose (54 mg, 0.364 mmol), AcOH (22 mg, 0.364 mmol) and NaBH.sub.3CN (22.5 mg, 0.364 mmol) then again heated to 45 C. and stirred for 24 h. The reaction mass concentrated under reduced pressure and the crude was purified by C-18 reverse phase combi flash purification eluting with 10-100% CH.sub.3CN in H.sub.2O (0.05% HCOOH) (compound isolated at 50% of CH.sub.3CN) to afford Exp-183b (300 mg, 27%) as a colourless gum: ESI (m/z) [C.sub.47H.sub.68N.sub.8O.sub.10+H].sup.+ 905.

[0956] Preparation of Exp-183c: To a solution of Exp-183b (300 mg, 0.331 mmol) in IPA (15 mL) and H.sub.2O (15 mL) was added 20% Pd(OH).sub.2 on carbon 50% wet (90 mg, 50% weight substrate) under inert atmosphere. The resultant reaction mixture stirred under H.sub.2 (balloon) for 16 h. The reaction mass filtered through a celite bed, and the filtrate was concentrated under reduced pressure. The obtained crude was azeotrope with CH.sub.3CN (210 mL) and dried under reduced pressure to afford Exp-183c (210 mg, 82%) as a colourless gum; ESI (m/z) [C.sub.39H.sub.62N.sub.8O.sub.8+H].sup.+ 771.

[0957] Preparation of Exp-183d: To a solution of Exp-183c (210 mg, 0.272 mmol) and compound 1 (116 mg, 0.299 mmol) in DMF (5 mL) was added DIPEA (0.13 mL, 0.816 mmol). The reaction mixture was heated to 60 C. and stirred for 16 h. The reaction mass was concentrated under reduced pressure, the obtained crude was stirred in IPA (210 mL), decanted and dried under reduced pressure to afford compound Exp-183d [280 (rude)] as a yellow gum: ESI (m/z) [C.sub.45H.sub.67ClN.sub.14O.sub.9+H].sup.+ 983.

[0958] Preparation of Exp-183: A stirred solution of Exp-183d (280 mg (crude)) in MeOH (5 mL) was charged with 4M HCl in MeOH (5.0 mL) at room temperature then stirred for 3 h. The reaction mixture was concentrated under reduced pressure. Obtained crude was purified by C-18 reverse phase combi-flash chromatography eluted with 10%-100% CH.sub.3CN in water (0.05% HCl), compound isolated at 35% of CH.sub.3CN to afford compound Exp-183 (57 mg, 23% over two steps) as a light yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.44-7.40 (m, 4H), 7.25-7.18 (m, 4H), 5.43-5.40 (m, 1H), 3.94-3.92 (m, 1H), 3.71-3.65 (m, 2H), 3.58-3.51 (m, 3H), 3.42-3.38 (m, 1H), 3.29-3.22 (m, 3H), 3.08-2.90 (m, 2H), 2.90-2.79 (m, 7H), 2.70-2.62 (m, 3H), 2.30-2.15 (m, 1H), 2.05-1.90 (m, 1H), 1.72-1.52 (m, 8H), 1.37-1.30 (m, 2H); ESI (m/z) [C.sub.40H.sub.59ClN.sub.14O.sub.7+H].sup.+ 884; HPLC AUC 93.3% (R.sub.t=7.31 min); HPLC, AUC=93.4%; t.sub.R=7.31 min, Method W.

Example 184

Preparation of 3,5-diamino-N(N-(4-(4-(3-(((S)-5-amino-1-(1H-tetrazol-5-yl)pentyl)(3-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)propyl)amino)-3-oxopropyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)-6-chloropyrazine-2-carboxamide (Exp-184)

##STR00543##

[0959] Preparation of Exp-184: Exp-184 was prepared in an analogous five step procedure to Exp-176 using similar conditions and reagents: .sup.1H NMR (400 MHz, CD.sub.3OD) 7.43-7.39 (m, 4H), 7.25-7.18 (m, 4H), 5.86-5.83 (in, 0.3H), 5.28-5.26 (m, 0.7H), 4.04-3.98 (m, 2H), 3.75-3.68 (m, 4H), 3.62-3.50 (m, 7H), 3.27-3.20 (m, 4H), 3.19-3.05 (i, 4H), 2.98-2.90 (5, 4H), 2.85-2.78 (m, 3H), 2.68-2.60 (m, 2H), 2.25-2.10 (m, 2H), 1.79-1.61 (m, 6H), 1.37-1.22 (m, 4H); ESI (m/z) [C.sub.46H.sub.71ClN.sub.14O.sub.12+H]1047; HPLC AUC 98.1% (R.sub.t=7.09 mmi); HPLC, AUC=98.1%; t.sub.R=7.09 min, Method W.

Examples 185-188

[0960] Preparation of Exp-185-188: In an analogous sequence of procedures used for Examples 179-182, Examples 185-188 were synthesized substituting D-lysine for L-lysine at the beginning of the reaction scheme:

##STR00544##

TABLE-US-00044 Exp R.sup.9 R R.sup.1 Spectra 185 CH.sub.3 Sug H .sup.1H NMR (400 MHz, CD.sub.3OD) 7.46-7.32 (m, 4H), 7.24-7.10 (m, 4H), 4.07-3.89 (m, 2H), 3.72-3.58 (m, 4H), 3.57-3.50 (m, 4H), 3.45-3.40 (m, 1H), 3.27-3.22 (m, 4H), 3.09-2.95 (m, 3H), 2.89-2.74 (m, 4H), 2.64-2.54 (m, 4H), 2.05-1.90 (m, 2H), 1.84-1.76 (m, 1H), 1.71-1.50 (m, 7H), 1.41-1.19 (m, 2H); ESI (m/z) [C.sub.43H.sub.63ClN.sub.8O.sub.9 + H].sup.+ 871; HPLC AUC 92.7 % (R.sub.t = 7.39 min); HPLC, AUC = 92.8%; t.sub.R = 7.39 min, Method W. 186 H Sug H .sup.1H NMR (400 MHz, CD.sub.3OD) 7.44-7.38 (m, 4H), 7.24-7.16 (m, 4H), 4.28-4.19 (m, 0.3H), 4.17-4.08 (m, 0.8H), 4.00-3.92 (m, 1H), 3.75-3.71 (m, 1H), 3.71-3.64 (m, 1H), 3.63-3.52 (m, 3H), 3.51-3.37 (m, 2H), 3.29-3.23 (m, 2H), 3.19-3.12 (m, 1H), 3.12-2.73 (m, 9H), 2.65-2.59 (m, 2H); 1.82-1.98 (m, 3H), 1.73-1.55 (m, 7H), 1.38-1.22 (m, 2H); ESI (m/z) [C.sub.40H.sub.59ClN.sub.10O.sub.9 + H].sup.+ 859; HPLC, AUC = 96.1%; t.sub.R = 7.15 min, Method A. 187 CH.sub.3 Sug Sug .sup.1H NMR (400 MHz, CD.sub.3OD) 7.48-7.37 (m, 4H), 7.27-7.16 (m, 4H), 4.19-4.01 (m, 2H), 3.80-3.72 (m, 2H), 3.71-3.62 (m, 4H), 3.61-3.54 (m, 7H), 3.44-3.30 (m, 2H), 3.29-3.24 (m, 3H), 3.24-3.21 (m, 4H), 3.08-3.02 (m, 1H), 2.93-2.72 (m, 5H), 2.89-2.61 (m, 4H), 2.11-1.97 (m, 2H), 1.75-1.54 (m, 8H), 1.46-1.22 (m, 2H); ESI (m/z) [C.sub.47H.sub.73ClN.sub.10O.sub.15 + H].sup.+ 1037; HPLC, AUC = 91.9%; t.sub.R = 7.27 min, Method W. 188 H Sug Sug .sup.1H NMR (400 MHz, CD.sub.3OD) 7.43-7.39 (m, 4H), 7.21-7.18 (m, 4H), 4.51-4.50 (m, 1H), 4.07-4.01 (m, 2H), 3.73-3.54 (m, 10H), 3.38-3.12 (m, 10H), 2.88-2.80 (m, 4H), 2.73-2.61 (m, 4H), 1.97-1.92 (m, 3H), 1.72-1.57 (m, 7H), 1.31-1.21 (m, 2H); ESI (m/z) [C.sub.46H.sub.71ClN.sub.10O.sub.14 + H].sup.+ 1023; HPLC AUC 96.5% (R.sub.t = 7.13 min); HPLC, AUC = 96.6%; t.sub.R = 7.13 min, Method W.

Example 189

Preparation of (2S,3R,4R,5R)N-(2-(3-(4-(4-(3-(3,5-diamino-6-chloropyrazine-2-carbonyl)guanidino)butyl)-[1,1-biphenyl]-4-yl)propanamido)ethyl)-2,3,4,5,6-pentahydroxy-N-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)hexan-1-amine oxide (Exp-189)

##STR00545##

[0961] Preparation of Exp-189: A suspension of Exp-87 (1.5 g, 1.704 mmol) in 3% H.sub.2O.sub.2 in H.sub.2O (6.0 mL, 5.112 mmol) was stirred at room temperature for 48 h. The reaction mass diluted with CH.sub.3CN (30 mL), stirred for 10 minutes and the solid filtered. The isolated solid was stirred in MTBE (220 mL), the solvent was decanted, filtered and then dried under reduced pressure for 3 h to afford compound Exp-189 (1.30 g, 85%) as an off white solid; .sup.1H NMR (400 MHz, DMSO-d.sub.6, (D.sub.2O exchange) 7.56-7.54 (m, 4H), 7.30-7.28 (m, 4H), 4.29-4.24 (m, 2H), 3.96-3.94 (m, 4H), 3.68-3.41 (m, 14H), 3.21-3.18 (m, 2H), 2.86 (t, J=7.6 Hz, 2H), 2.64-2.54 (m, 2H), 2.45 (t, J=7.6 Hz, 2H), 1.68-1.52 (m, 4H); ESI (m/z) [C.sub.39H.sub.58ClN.sub.9O.sub.13+H].sup.+ 896; HPLC AUC 91.6% (R.sub.t=6.41 min); Method Z.

Example 190

Preparation of 3,5-diamino-6-chloro-N(N-(4-(4-(3-oxo-3-((2-(((2R,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl) amino) ethyl) amino) propyl)-[1,1-biphenyl]-4-yl)butyl)carbamimidoyl)pyrazine-2-carboxamide (Exp-190)

##STR00546##

[0962] Preparation of Exp-190: In a three-step procedure Exp-190 was synthesized beginning with Int-23 and Int-60 using a sequence analogous to the one used to prepare Exp-87: .sup.1H NMR (400 MHz, DMSO-d.sub.6, D.sub.2O exchange) 7.55-7.53 (m, 4H), 7.29-7.26 (m, 4H), 3.74-3.70 (m, 1H), 3.69-3.45 (m, 9H), 3.44-3.41 (m, 2H), 3.18-3.15 (m, 4H), 2.85-2.80 (m, 3H), 2.66-2.58 (m, 5H), 2.48-2.38 (m, 4H), 1.66-1.63 (m, 2H), 1.56-1.54 (m, 2H); ESI (m/z) [C.sub.39H.sub.58ClN.sub.9O.sub.12+H].sup.+ 880; HPLC AUC 96.3% (R.sub.t=2.77 min); UPLC method AB.

[0963] Several assays may be used to characterize the compounds of the present invention. Representative assays are discussed below.

In Vitro Measure of Sodium Channel Blocking Activity

[0964] One assay used to assess mechanism of action and/or potency of the compounds of the present invention involves the determination of lumenal drug inhibition of sodium current measured in airway epithelial monolayers mounted in Ussing-type chambers. Cells derived from excised human lung tissue are seeded onto porous (0.4 micron) SnapWell inserts (CoStar), cultured under air-liquid interface (ALI) conditions in hormonally defined media, and assayed for electrogenic sodium transport while submerged in Krebs bicarbonate Ringer (KBR) solution. Addition of test drug is made to the lumenal bath chamber in half-log dose increments (range: 110.sup.11 M to 410.sup.6 M), and the cumulative change in short-circuit current (Isc, mAmps/cm.sup.2) is recorded. All drugs are prepared in dimethyl sulfoxide as stock solutions at a concentration of 110.sup.2 M and stored at 20 C. until use. Six preparations are typically run in parallel. Analog Isc output from a commercial voltage-clamp amplifier (Physiologic Instruments, Reno, NV) is digitized and acquired via computer using data acquisition software (Physiologic Instruments).

[0965] The inhibitor concentration achieving 50% Isc blockade (IC5O) is obtained from model fits (SigmaPlot v13, SYSTAT, San Jose, CA) to the dose-response relation for each of the normalized ENaC blocker-sensitive Isc measurements. Selected sodium channel blocker IC50s are presented in Table E1 as an index of potency.

Plasma Protein Binding (PPB) Assay for ENaCi

[0966] The objective of this experiment is to use equilibrium dialysis to measure the free and protein-bound fractions of ENaCi in plasma. Briefly, 2 mM ENaCi stocks in DMSO are spiked into normal pooled human plasma (BioIVT). Final concentrations of 2 M ENaC blocker in plasma are transferred to a perfusate chamber of RED device (Thermofisher #89809) and dialyzed against Dulbeccos PBS (Sigma). Replicates are run in triplicate on the RED device plate at 37 C. with rotational agitation. Following incubation for 4 h, samples from both chambers of the RED device are assayed for ENaC blocker content by UPLC-FLR. Additional authentic standards are spiked into plasma between 2-0.02 M and both standards and assay samples are bioanalytically worked-up by protein precipitation with 3 acetonitrile+0.1% formic acid. Following centrifugation, supernatant is diluted 3 with water+0.1% formic acid. Finally, samples and standards are chromatographed on a Waters Acquity UPLC using a linear gradient of acetonitrile and water+0.1% trifluoroacetic acid. Data is collected and processed with Empower 3 software.

[0967] After interpolation from standard curve, the measured concentrations of ENaC blocker are used to calculate % free and % bound as follows:

[00001]$\begin{array}{c}\%\mathrm{free}=\left(\mathrm{concentration}\mathrm{buffer}\mathrm{chamber}/\mathrm{concentration}\mathrm{perfusate}\mathrm{chamber}\right)100\%\\ \%\mathrm{bound}=100\%-\%\mathrm{free}\end{array}$

[0968] Selected ENaCi PPB values are presented in Table E1.

TABLE-US-00045 TABLE E1 IC.sub.50 (nM); PPB Data Example # IC.sub.50 (nM) % Protein Binding 1 5.98 99.9 2 0.92 99.4 3 39.46 NT 4 0.48 99.3 5 9.23 NT 6 0.279 99.5 7 1.32 NT 8 23.01 NT 9 3.23 NT 10 6.11 99.3 11 4.63 NT 12 39.46 99.5 13 3.14 99.6 14 146.71 NT 15 23.496 NT 16 3.87 99.9 17 2.45 99.6 18 0.193 95.2 19 1.45 95.2 20 0.234 99.3 21 1.90 96.3 22 0.489 98.3 23 3.70 94.5 24 0.214 96.2 25 1.52 96.6 26 0.44 NT 27 1.97 96.8 28 1.78 99.8 29 38.52 99.4 30 0.52 99.7 31 8.04 96.2 32 1.06 99.8 33 4.69 99.7 34 1.96 99.0 35 7.45 98.4 36 0.46 97.5 37 5.25 95.0 38 0.251 NT 39 2.80 NT 40 0.91 NT 41 4.322 NT 42 0.40 98.5 43 2.45 94.8 44 0.907 NT 45 4.63 NT 46 0.76 NT 47 7.71 NT 48 0.25 98.6 49 2.06 91.0 50 0.864 97.3 51 1.24 97.5 52 4.98 NT 53 0.22 99.5 54 1.56 94.7 55 1.59 NT 56 1.02 NT 57 2.68 98.9 58 0.501 NT 59 27.58 96.1 60 15.97 NT 61 0.827 NT 62 44.75 NT 63 1.63 93.1 64 3.31 92.3 65 0.944 95.1 66 7.36 92.0 67 2.51 >99.9 68 7.04 99.9 69 1.425 99.3 70 1.68 98.9 71 0.80 99.2 72 1.39 97.8 73 0.807 98.7 74 0.401 NT 75 0.809 NT 76 0.499 NT 77 1.34 NT 78 0.831 NT 79 1.06 NT 80 0.414 NT 81 0.918 NT 82 0.576 NT 83 0.930 NT 84 1.11 NT 85 0.474 95.6 86 1.28 98.9 87 1.40 98.5 88 0.682 NT 89 1.46 NT 90 1.52 99.4 91 2.90 94.0 92 0.998 98.8 93 54.385 NT 94 10.67 NT 95 1.30 97.6 96 9.86 NT 97 0.96 99.4 98 1.13 96.6 99 0.69 98.5 100 1.90 NT 101 2.15 >99.9 102 2.80 98.7 103 2.80 97.1 104 2.16 93.0 105 0.203 98.1 106 3.12 90.4 107 0.361 96.4 108 4.92 87.8 109 0.395 NT 110 1.82 NT 111 0.281 NT 112 6.97 NT 113 0.448 97.5 114 5.42 92.1 115 0.68 NT 116 11.62 92.2 117 0.18 98.3 118 0.47 98.1 119 0.402 NT 120 0.35 97.8 121 2.34 91.5 122 0.637 97.0 123 7.15 93.0 124 0.506 NT 125 0.768 NT 126 0.291 98.3 127 3.14 82.1 128 0.44 NT 129 1.97 96.8 130 0.361 NT 131 0.464 NT 132 4.66 NT 133 1.35 99.9 134 6.42 90.8 135 0.209 98.5 136 0.23 98.1 137 0.406 NT 138 0.508 99.7 139 15.81 86.9 140 1.40 98.5 141 0.539 95.7 142 0.611 99.5 143 .805 95.6 144 1.45 96.7 145 1.14 97.5 146 0.69 99.1 147 0.64 NT 148 2.89 NT 149 2.00 99.4 150 5.56 95.1 151 0.68 99.2 152 1.51 96.1 153 3.07 97.7 154 1.71 96.4 155 113.18 NT 156 13.55 NT 157 15.29 NT 158 0.18 98.3 159 1.18 99.0 160 1.81 NT 161 4.73 97.6 162 2.22 98.3 163 2.82 NT 164 6.70 NT 165 2.34 97.0 166 3.54 NT 167 6.88 NT 168 1.54 98.7 169 2.29 NT 170 5.33 NT 171 1.90 93.9 172 0.17 97.1 173 0.39 93.1 174 0.672 91.3 175 4.81 86.0 176 0.715 NT 177 0.91 NT 178 5.96 NT 179 0.145 96.9 180 0.79 95.1 181 0.34 85.2 182 3.70 81.9 183 1.24 NT 184 2.35 89.05 185 0.43 NT 186 0.62 92.2 187 0.73 NT 188 2.79 NT 189 2.34 NT 190 1.19 NT NT = Not tested

EQUIVALENTS AND SCOPE

[0969] In the claims articles such as a, an, and the may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include or between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The invention includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The invention includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process.

[0970] Furthermore, the invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the invention, or aspects of the invention, is/are referred to as comprising particular elements and/or features, certain embodiments of the invention or aspects of the invention consist, or consist essentially of, such elements and/or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms comprising and containing are intended to be open and permits the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0971] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present invention that falls within the prior art may be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the invention can be excluded from any claim, for any reason, whether or not related to the existence of prior art.

[0972] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description, but rather is as set forth in the appended claims. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present invention, as defined in the following claims.