SMALL MOLECULE ANTAGONIST TO PACAP RECEPTOR AND USES THEREOF

Abstract

The invention relates to compounds that demonstrate efficacy in blocking PACAP receptors (e.g. PAC1) and associated methods of treating neurological diseases or disorders using the disclosed compounds. The neurological diseases or disorders include, but are not limited to, stress-related diseases (e.g. chronic stress, post-traumatic stress disorder (PTSD), anxiety, or general anxiety disorder), pain-related diseases (e.g. chronic pain, primary headache disorders, or migraines), or addiction (e.g. addiction to a substance (e.g. addiction to cocaine, amphetamines, methamphetamine, methylphenidate, nicotine, alcohol, prescription medication, marijuana, tobacco, or an opioid selected from heroin, fentanyl, codeine, hydrocodone, morphine, oxycodone, hydromorphone, and methadone)).

Claims

1. A compound of Formula (I): ##STR00052## or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, isotopologue, stereoisomer, or prodrug thereof, wherein: R.sup.1 is independently optionally substituted aryl, optionally substituted heteroaryl, halogen, —CN, —NO.sub.2, or —C(═O)OR.sup.a, wherein each Ra is independently H, or optionally substituted C.sub.1-6 alkyl; R.sup.2 is halogen or —NO.sub.2; X.sup.1 and X.sup.2 are independently C—H or N; and n is 1, 2, or 3.

2. The compound of claim 1, wherein X.sup.1 is C—H.

3. The compound of claim 1, wherein X.sup.1 is N.

4. The compound of any one of claims 1-3 or, wherein X.sup.2 is C—H.

5. The compound of any one of claim 1 or 2, wherein X.sup.2 is N.

6. The compound of any one of claims 1-5, wherein n is 1.

7. The compound of any one of claims 1-5, wherein n is 2.

8. The compound of any one of claims 1-7, wherein at least one instance of R.sup.1 is —NO.sub.2.

9. The compound of any one of claims 1-7, wherein at least one instance of R.sup.1 is —CN.

10. The compound of any one of claims 1-7, wherein at least one instance of R.sup.1 is a halogen.

11. The compound of claim 10, wherein at least one instance of R.sup.1 is —F.

12. The compound of any one of claims 1-7, wherein at least one instance of R.sup.1 is —C(═O)OR.sup.a.

13. The compound of claim 12, wherein Ra is unsubstituted alkyl.

14. The compound of claim 13, wherein Ra is unsubstituted methyl.

15. The compound of claim 14, wherein Ra is H.

16. The compound of any one of claims 1-7, wherein R.sup.1 is optionally substituted aryl.

17. The compound of claim 16, wherein R.sup.1 is phenyl.

18. The compound of claim 16 or 17, wherein R.sup.1 is unsubstituted phenyl.

19. The compound of any one of claims 1-7, wherein R.sup.1 is optionally substituted heteroaryl.

20. The compound of claim 19, wherein R.sup.1 is optionally substituted pyrimidine.

21. The compound of claim 19, wherein R.sup.1 is unsubstituted pyrimidine.

22. The compound of any one of claims 1-21, wherein R.sup.2 is NO.sub.2.

23. The compound of any one of claims 1-22, of the formula: ##STR00053## or a salt thereof.

24. A compound of Formula (II): ##STR00054## or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, isotopologue, stereoisomer, or prodrug thereof, wherein: R.sup.7 is halogen, —CF.sub.3, —CN, —NO.sub.2, or —C(═O)OR.sup.c, wherein each R.sup.c is independently optionally substituted C.sub.1-6 alkyl.

25. The compound of claim 24, wherein R.sup.7 is —CF.sub.3.

26. The compound of any one of claim 24 or 25, of the formula: ##STR00055## or a salt thereof.

27. A compound of Formula (III): ##STR00056## or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, isotopologue, stereoisomer, or prodrug thereof, wherein: R.sup.3 is hydrogen, halogen, —CF.sub.3, —CN, —NO.sub.2, —C(═O)OR.sup.c, wherein each R.sup.c is independently optionally substituted C.sub.1-6 alkyl; R.sup.4 and R.sup.5 are independently halogen, —CF.sub.3, —CN, —NO.sub.2, or —C(═O)OR.sup.b, wherein R.sup.b is optionally substituted C.sub.1-6 alkyl; R.sup.6 is optionally substituted aryl, optionally substituted C.sub.1-6 alkyl, or optionally substituted C.sub.1-8 heteroalkyl; and X is C—H or N.

28. The compound of claim 27, wherein X is C—H.

29. The compound of claim 27, wherein X is N.

30. The compound of any one of claims 27-29, wherein R.sup.2 is —CF.sub.3.

31. The compound of claim 30, wherein R.sup.4 is —CF.sub.3.

32. The compound of any one of claims 27-29, wherein R.sup.2 is a halogen.

33. The compound of any one of claims 27-29, wherein R.sup.2 is —F.

34. The compound of claim 32 or 33, wherein R.sup.4 is a halogen.

35. The compound of any one of claims 32-34, wherein R.sup.4 is —F.

36. The compound of any one of claims 27-35, wherein R.sup.6 is optionally substituted alkyl.

37. The compound of any one of claims 27-36, wherein R.sup.6 is unsubstituted alkyl.

38. The compound of claim 37, wherein R.sup.6 is unsubstituted n-butyl.

39. The compound of any one of claims 27-35, wherein R.sup.6 is optionally substituted heteroalkyl.

40. The compound of any one of claims 27-39, of the formula: ##STR00057## or a salt thereof.

41. A compound of Formula (IV): ##STR00058## or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, isotopologue, stereoisomer, or prodrug thereof, wherein: R.sup.8 is —C(═O)OR.sup.d, —C(═O)N(R.sup.d).sub.2 wherein each R.sup.d is independently H, or optionally substituted C.sub.1-6 alkyl.

42. The compound of claim 41, wherein R.sup.8 is —C(═O)OR.sup.d.

43. The compound of claim 42, wherein R.sup.d is optionally substituted alkyl.

44. The compound of claim 42 or 43, wherein R.sup.d is ethyl.

45. A pharmaceutical composition comprising a compound of any one of claims 1-44, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, isotopologue, stereoisomer, or prodrug thereof, and optionally a pharmaceutically acceptable excipient.

46. The pharmaceutical composition of claim 45 further comprising an additional pharmaceutical agent.

47. A method of treating a subject suffering from or susceptible to a neurological disease, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00059## or a salt thereof.

48. The method of claim 47, wherein the neurological disease is a pain-related disorder.

49. The method of claim 47, wherein the neurological disorder is a stress-related disorder.

50. The method of claim 47, wherein the neurological disorder is addiction.

51. The method of claim 47, wherein the neurological disorder is a panic disorder.

52. The method of claim 47, wherein the neurological disorder is panic attacks.

53. The method of claim 47, wherein the neurological disorder is an eating disorder.

54. A method of treating a subject suffering from or susceptible to a pain-related disease, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00060## or a salt thereof.

55. The method of claim 54, wherein the pain-related disease is chronic pain.

56. The method of claim 54, wherein the pain-related disease is a primary headache disorder.

57. The method of claim 54, wherein the primary headache disorder is a migraine.

58. A method of treating a subject suffering from or susceptible to a stress-related disease or disorder, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00061## or a salt thereof.

59. The method of claim 58, wherein the stress-related disorder is chronic stress.

60. The method of claim 58, wherein the stress-related disorder is PTSD.

61. The method of claim 58, wherein the stress-related disorder is an anxiety disorder.

62. The method of claim 58, wherein the stress-related disorder is anxiety.

63. The method of claim 58, wherein the anxiety disorder is generalized anxiety disorder.

64. A method of treating a subject suffering from or susceptible to addiction, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00062## ##STR00063## or a salt thereof.

65. The method of claim 64, wherein the addiction is addiction to a substance.

66. The method of claim 65, wherein the substance is a drug.

67. The method of claim 65, wherein the substance is food.

68. The method of claim 65, wherein the substance is cocaine, amphetamines, methamphetamine, methylphenidate, nicotine, alcohol, prescription medication, marijuana, tobacco, or an opioid selected from heroin, fentanyl, codeine, hydrocodone, morphine, oxycodone, hydromorphone, or methadone.

69. The method of claim 65, wherein the substance is cocaine.

70. The method of claim 65, wherein the substance is an opioid.

71. The method of claim 65. wherein the substance is an amphetamine.

72. The method of claim 65, wherein the substance is nicotine.

73. The method of claim 65, wherein the substance is alcohol.

74. The method of any one of claims 64-73, wherein the treatment prevents relapse of the substance abuse.

75. The method of any one of claims 64-73, wherein the treatment lowers the incidence of relapse of the substance abuse.

76. The method of claim 74 or 75, wherein the relapse is stress-induced relapse.

77. A method of treating a subject suffering from or susceptible to an eating disorder, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00064## ##STR00065## or a salt thereof.

78. The method of claim 77, wherein the eating disorder is orthorexia.

79. The method of claim 77, wherein the eating disorder is anorexia.

80. A method of treating a subject identified as in need of treatment for a pain-related disease or disorder, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00066## ##STR00067## or a salt thereof.

81. A method of treating a subject identified as in need of treatment for a stress-related disorder, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00068## ##STR00069## or a salt thereof.

82. A method of treating a subject identified as in need of treatment for a panic disorder, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00070## ##STR00071## or a salt thereof.

83. A method of treating a subject identified as in need of treatment for addiction, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00072## ##STR00073## or a salt thereof.

84. A method of treating a subject identified as in need of treatment for an eating disorder, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00074## ##STR00075## or a salt thereof.

85. The method of any one of claims 47-84 further comprising administering an additional pharmaceutical agent to the subject.

86. The method of claim 85, wherein the additional pharmaceutical agent is a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00076## ##STR00077## or a salt thereof.

87. The method of claim 85, wherein the additional pharmaceutical agent is the compound I-20, or a salt thereof.

88. A kit comprising: a compound of any one of claims 1-44, a pharmaceutical composition of claim 45 or 46, or a compound of the formula: ##STR00078## ##STR00079## or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or stereoisomer thereof; and instructions for administering to a subject or contacting a biological sample with the compound, or composition.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0056] FIG. 1 depicts treatment of HEK cells treated with PACAP and PACAP/compound I-1.

[0057] FIG. 2 depicts the F340/F380 kinetic ratio kinetic of the concentration of Ca2+ in HEK cells treated with PACAP or PACAP/compound I-1.

[0058] FIG. 3 depicts compound 1 blocks PAC1 receptor-mediated ERK activation in a dose-dependent manner using the HEK PAC1-EGFP receptor cell line.

[0059] FIG. 4 depicts attenuated ERK activation using compound I-1.

[0060] FIG. 5 depicts attenuated ERK activation using compound I-20.

[0061] FIG. 6 depicts attenuated ERK activation using compounds I-1 and I-20.

[0062] FIG. 7 depicts comparisons of different acyl hydrazides in blocking PACAP-stimulated ERK activation.

[0063] FIG. 8 depicts open field tests on male rats with BNST PACAP infusions, with or without compound I-1 pre-administration.

[0064] FIG. 9 depicts the effect of compound I-5 on PACAP-stimulated calcium signaling in the PAC1-EGFP HEK cell lines.

[0065] FIG. 10 depicts the effect of compound I-6 on PACAP-stimulated calcium signaling in the PAC1-EGFP HEK cell lines.

[0066] FIG. 11 depicts the effect of compound I-7 on PACAP-stimulated calcium signaling in the PAC1-EGFP HEK cell lines.

[0067] FIG. 12 depicts the effect of compound I-2 on PACAP-stimulated calcium signaling in the PAC1-EGFP HEK cell lines.

[0068] FIG. 13 depicts the effect of compound I-22 on PACAP-stimulated calcium signaling in the PAC1-EGFP HEK cell lines.

[0069] FIG. 14 depicts the effect of compound I-3 on PACAP-stimulated calcium signaling in the PAC1-EGFP HEK cell lines.

[0070] FIG. 15 depicts the effect of compound I-18 on PACAP-stimulated calcium signaling in the PAC1-EGFP HEK cell lines.

[0071] FIG. 16 depicts the effect of compound I-19 on PACAP-stimulated calcium signaling in the PAC1-EGFP HEK cell lines.

[0072] FIG. 17 depicts the effect of compound I-21 on PACAP-stimulated calcium signaling in the PAC1-EGFP HEK cell lines.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION

[0073] The present invention provides compounds that act as antagonists for a PACAP-receptor (e.g. PAC1), and pharmaceutical compositions thereof, for the prevention and treatment of a disease or disorder in a subject. In certain embodiments, the compounds act as antagonists to PACAP receptors (e.g. PAC1). In certain embodiments, the compounds are irreversible antagonist for PACAP receptors (e.g. PAC1). In certain embodiments, the compounds are reversible antagonist for PACAP receptors (e.g. PAC1). The present invention further provides methods of using the compounds described herein, e.g., as biological probes to study the antagonism of PACAP receptors (e.g. PAC1), and as therapeutics, e.g., in the prevention and treatment of diseases or disorders associated with PACAP receptor activity or internalization, or ERK activation. In certain embodiments, the diseases or disorders include, but are not limited to, neurological disorders, e.g. stress-related disorders (e.g. chronic stress, post-traumatic stress disorder (PTSD), anxiety, or general anxiety disorder), eating disorders (e.g. orthorexia, or anorexia nervosa), pain-related disorders (e.g. chronic pain, primary headache disorders, or migraines), panic disorders, panic attacks, or addiction (e.g. addiction to a substance (e.g. addiction to cocaine, amphetamines, methamphetamine, methylphenidate, nicotine, alcohol, prescription medication, marijuana, tobacco, food, or an opioid selected from heroin, fentanyl, codeine, hydrocodone, morphine, oxycodone, hydromorphone, and methadone)).

Compounds

[0074] In one aspect of the present invention, provided are compounds of Formula (I):

##STR00002##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, isotopologue, stereoisomer, or prodrug thereof, wherein:

[0075] R.sup.1 is independently optionally substituted aryl, optionally substituted heteroaryl, halogen, —CN, —NO.sub.2, or —C(═O)OR.sup.a, wherein each Ra is independently H, or optionally substituted C.sub.1-6 alkyl;

[0076] R.sup.2 is halogen or —NO.sub.2;

[0077] X.sup.1 and X.sup.2 are independently C—H or N; and

[0078] n is 1, 2, or 3.

[0079] Formula (I) contains the substituent R.sup.1. Formula (I) contains n instances of R.sup.1, wherein n is an integer between 1 and 3. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments n is 3. In certain embodiments, at least one instance of R.sup.1 is a halogen. In certain embodiments, at least one instance of R.sup.1 is F. In certain embodiments, at least two instances of R.sup.1 are F. In certain embodiments n is 2, and R.sup.1 is F. In certain embodiments, at least one instance of R.sup.1 is —CN. In certain embodiments, at least one instance of R.sup.1 is —NO.sub.2. In certain embodiments, at least one instance of R.sup.1 is —C(═O)OR.sup.a, wherein Ra is optionally substituted C.sub.1-6 alkyl (e.g. Me, Et, n-Pr). In certain embodiments, at least one instance of R.sup.1 is CO.sub.2Me. In certain embodiments, at least one instance of R.sup.1 is optionally substituted aryl. In certain embodiments, at least one instance of R.sup.1 is unsubstituted aryl. In certain embodiments, at least one instance of R.sup.1 is unsubstituted phenyl. In certain embodiments, at least one instance of R.sup.1 is optionally substituted heteroaryl. In certain embodiments, R.sup.1 is optionally substituted pyridine, pyrimidine, pyridazine, pyrazine, or triazine. In certain embodiments, R.sup.1 is optionally substituted pyrimidine.

[0080] Formula (I) contains the substituent X.sup.1. In certain embodiments, X.sup.1 is C. In certain embodiments, X.sup.1 is N. In certain embodiments, X.sup.1 is N and at least one instance of R.sup.1 is —CN. In certain embodiments, X.sup.1 is C, and at least one instance of R.sup.1 is —CN. In certain embodiments, X.sup.1 is C, and at least one instance of R.sup.1 is —NO.sub.2. In certain embodiments, X.sup.1 is C, and at least one instance of R.sup.1 is a halogen. In certain embodiments, X.sup.1 is C, and at least one instance of R.sup.1 is F. In certain embodiments, X.sup.1 is C, and at least one instance of R.sup.1 is —C(═O)OR.sup.a. In certain embodiments, X.sup.1 is C, and at least one instance of R.sup.1 is F. In certain embodiments, X.sup.1 is C, and at least one instance of R.sup.1 is —CO.sub.2Me.

[0081] Formula (I) contains the substituent X.sup.2. In certain embodiments, X.sup.2 is C. In certain embodiments, X.sup.2 is N. In certain embodiments, X.sup.2 is N and at least one instance of R.sup.1 is —CN. In certain embodiments, X.sup.2 is C, and at least one instance of R.sup.1 is —CN. In certain embodiments, X.sup.2 is C, and at least one instance of R.sup.1 is —NO.sub.2. In certain embodiments, X.sup.2 is C, and at least one instance of R.sup.1 is a halogen. In certain embodiments, X.sup.2 is C, and at least one instance of R.sup.1 is F. In certain embodiments, X.sup.2 is C, and at least one instance of R.sup.1 is —C(═O)OR.sup.a. In certain embodiments, X.sup.2 is C, and at least one instance of R.sup.1 is F. In certain embodiments, X.sup.2 is C, and at least one instance of R.sup.1 is —CO.sub.2Me.

[0082] In certain embodiments, Formula (I) is of the Formula (I-a):

##STR00003##

or pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, isotopologues, stereoisomers, or prodrugs thereof, wherein R.sup.1 is independently optionally substituted aryl, optionally substituted heteroaryl, halogen, —CN, —NO.sub.2, or —C(═O)OR.sup.a, wherein each Ra is independently H, or optionally substituted C.sub.1-6 alkyl.

[0083] In certain embodiments, Formula (I) is of the Formula (I-b):

##STR00004##

[0084] In certain embodiments, Formula (I) is of the Formula (I-c):

##STR00005##

[0085] In certain embodiments, Formula (I) is of the Formula (I-d):

##STR00006##

[0086] In certain embodiments, Formula (I) is of the Formula (I-e):

##STR00007##

[0087] In certain embodiments, a compound of Formula (I) is of the formula:

##STR00008##

or a salt thereof.

[0088] In one aspect of the present invention, provided are compounds of Formula (II):

##STR00009##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, isotopologue, stereoisomer, or prodrug thereof, wherein:

[0089] R.sup.7 is halogen, —CF.sub.3, —CN, —NO.sub.2, or —C(═O)OR.sup.c, wherein each R.sup.c is independently optionally substituted C.sub.1-6 alkyl.

[0090] Formula (II) contains the substituent R.sup.7. In certain embodiments, R.sup.7 is halogen. In certain embodiments, R.sup.7 is —CF.sub.3. In certain embodiments, R.sup.7 is —NO.sub.2. In certain embodiments, R.sup.7 is —C(═O)OR.sup.c, wherein each Reis independently optionally substituted C.sub.1-6 alkyl (e.g., Me, Et, n-Pr).

[0091] In certain embodiments, Formula (II) is of the formula:

##STR00010##

or a salt thereof.

[0092] In one aspect of the present invention, provided are compounds of Formula (III):

##STR00011##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, isotopologue, stereoisomer, or prodrug thereof, wherein:

[0093] R.sup.3 is hydrogen, halogen, —CF.sub.3, —CN, —NO.sub.2, —C(═O)OR.sup.c, wherein each R.sup.c is independently optionally substituted C.sub.1-6 alkyl;

[0094] R.sup.4 and R.sup.5 are independently halogen, —CF.sub.3, —CN, —NO.sub.2, or —C(═O)OR.sup.b, wherein R.sup.b is optionally substituted C.sub.1-6 alkyl;

[0095] R.sup.6 is optionally substituted aryl, optionally substituted C.sub.1-6 alkyl, or optionally substituted C.sub.1-8 heteroalkyl; and

[0096] X is C—H or N.

[0097] Formula (III) contains the substituent R.sup.3. In certain embodiments, R.sup.3 is H. In certain embodiments, R.sup.3 is halogen. In certain embodiments, R.sup.3 is F. In certain embodiments, R.sup.3 is —CF.sub.3. In certain embodiments, R.sup.3 is —CN. In certain embodiments, R.sup.3 is —NO.sub.2. In certain embodiments, R.sup.3 is —C(═O)OR.sup.c, wherein R.sup.c is optionally substituted C.sub.1-6 alkyl (e.g., Me, Et, n-Pr). In certain embodiments, R.sup.3 is C(═O)OMe.

[0098] Formula (III) contains the substituent R.sup.4. In certain embodiments, R.sup.4 is halogen. In certain embodiments, R.sup.4 is F. In certain embodiments, R.sup.4 is —CF.sub.3. In certain embodiments, R.sup.4 is —CN. In certain embodiments, R.sup.4 is —NO.sub.2. In certain embodiments, R.sup.4 is —C(═O)OR.sup.b, wherein R.sup.b is optionally substituted C.sub.1-6 alkyl (e.g., Me, Et, n-Pr).

[0099] Formula (III) contains the substituent R.sup.5. In certain embodiments, R.sup.5 is halogen. In certain embodiments, R.sup.5 is F. In certain embodiments, R.sup.5 is —CF.sub.3. In certain embodiments, R.sup.5 is —CN. In certain embodiments, R.sup.5 is —NO.sub.2. In certain embodiments, R.sup.5 is —C(═O)OR.sup.b, wherein R.sup.b is optionally substituted C.sub.1-6 alkyl (e.g., Me, Et, n-Pr).

[0100] Formula (III) contains the substituent X. In certain embodiments, X is C—H. In certain embodiments, X is N. In certain embodiments, X is C—H, R.sup.5 is —CF.sub.3, and R.sup.4 is —CF.sub.3. In certain embodiments, X is C—H, R.sup.5 is a halogen, and R.sup.4 is a halogen. In certain embodiments, X is C—H, R.sup.5 is —F, and R.sup.4 is a halogen. In certain embodiments, X is C—H, R.sup.2 is a halogen, and R.sup.4 is F. In certain embodiments, X is C—H, R.sup.5 is —F, and R.sup.4 is —F.

[0101] In certain embodiments, Formula (III) is of the Formula (III-a):

##STR00012##

or pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, isotopologues, stereoisomers, or prodrugs thereof, wherein R.sup.4 and R.sup.5 are independently halogen, —CF.sub.3, —CN, —NO.sub.2, or —C(═O)OR.sup.b, wherein each R.sup.b is independently optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.4 and R.sup.5 are the same. In certain embodiments, R.sup.4 and R.sup.5 are different.

[0102] In certain embodiments, Formula (III) is of the Formula (III-b):

##STR00013##

or pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, isotopologues, stereoisomers, or prodrugs thereof, wherein R.sup.3 is hydrogen, halogen, —CF.sub.3, —CN, —NO.sub.2, or —C(═O)OR.sup.c, wherein R.sup.c is optionally substituted C.sub.1-6 alkyl (e.g. Me, Et, n-Pr).

[0103] Formula (III-b) contains the substituent R.sup.3. In certain embodiments, R.sup.3 is H. In certain embodiments, R.sup.3 is halogen. In certain embodiments, R.sup.3 is F. In certain embodiments, R.sup.3 is —CF.sub.3. In certain embodiments, R.sup.3 is —CN. In certain embodiments, R.sup.3 is —NO.sub.2. In certain embodiments, R.sup.3 is —C(═O)OR.sup.c, wherein Reis optionally substituted C.sub.1-6 alkyl (e.g., Me, Et, n-Pr). In certain embodiments, R.sup.3 is C(═O)OMe.

[0104] In certain embodiments, a compound of Formula (III) is of the formula:

##STR00014##

or a salt thereof.

[0105] In one aspect of the present invention, provided are compounds of Formula (IV):

##STR00015##

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, isotopologue, stereoisomer, or prodrug thereof, wherein:

[0106] R.sup.8 is —C(═O)OR.sup.d, —C(═O)N(R.sup.d).sub.2 wherein each R.sup.d is independently H, or optionally substituted C.sub.1-6 alkyl.

[0107] Formula (IV) contains the substituent R.sup.8. In certain embodiments, R.sup.8 is —C(═O)OR.sup.d, —C(═O)N(R.sup.d).sub.2 wherein each R.sup.d is independently H, or optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.8 is CO.sub.2Et.

[0108] In certain embodiments, Formula (IV) is of the formula:

##STR00016##

or a salt thereof.

[0109] In certain embodiments, a compound described herein is any of the compounds I-1 to I-17, found in Table 1, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopologue, or prodrug thereof. In certain embodiments, a compound described herein is any of the compounds I-1 to I-17, or a pharmaceutically acceptable salt thereof.

TABLE-US-00001 TABLE 1 Compounds I-1 to I-17 of the disclosure Compound Number: Structure: I-1 [00017] embedded image I-2 [00018] I-3 [00019] I-4 [00020] I-5 [00021] I-6 [00022] I-7 [00023] I-8 [00024] I-9 [00025] I-10 [00026] I-11 [00027] I-12 [00028] I-13 [00029] I-14 [00030] I-15 [00031] I-16 [00032] I-17 [00033]

Methods of Treatment and Uses

[0110] The compounds described herein may be useful in treating and/or preventing diseases or disorders including, but not limited to, addiction (e.g. addiction to a substance (e.g. addiction to cocaine, amphetamines, methamphetamine, methylphenidate, nicotine, alcohol, prescription medication, marijuana, tobacco, food, or an opioid selected from heroin, fentanyl, codeine, hydrocodone, morphine, oxycodone, hydromorphone, and methadone)), stress-related disorders (e.g. chronic stress, PTSD, anxiety, or general anxiety disorders), eating disorders (e.g. orthorexia, or anorexia nervosa), or pain-related diseases or disorders (e.g. chronic pain, primary headache disorders, or migraines), or diseases and disorders associated with the activity of a PACAP receptor in a subject, or modulating the activity of a PACAP receptor in a subject, biological sample, tissue, or cell.

[0111] In one aspect, the invention provides a method of treating a subject suffering from or susceptible to a neurological disorder (e.g. stress-related disorders (e.g. chronic stress, post-traumatic stress disorder (PTSD), anxiety, or general anxiety disorder), eating disorders (e.g. orthorexia, or anorexia nervosa), pain-related disorders (e.g. chronic pain, primary headache disorders, or migraines), panic disorders, panic attacks, or addiction (e.g. addiction to a substance (e.g. addiction to cocaine, amphetamines, methamphetamine, methylphenidate, nicotine, alcohol, prescription medication, marijuana, tobacco, food, or an opioid selected from heroin, fentanyl, codeine, hydrocodone, morphine, oxycodone, hydromorphone, and methadone)), the method comprising administering to the subject in need thereof a therapeutically effective amount of any one of the compounds of Formula (I), Formula (II), or compounds I-18 to I-26 (see Table 2), or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopologue, or prodrug thereof. In certain aspects, the method treats addiction by preventing relapse of an addiction. In another aspect, the method treats addiction by lowering the incidence of relapse, as compared to a control population not receiving treatment via the method. In certain aspects, the method treats addiction by preventing stress-induced relapse of an addiction.

TABLE-US-00002 TABLE 2 Compounds I-18 to I-26 Compound Number: Structure: I-18 [00034] embedded image I-19 [00035] I-20 [00036] I-21 [00037] I-22 [00038] I-23 [00039] I-24 [00040] I-25 [00041] I-26 [00042]

[0112] In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-1, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-2, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-3, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-4, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-5, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-6, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-7, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-8, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-9, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-10, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-11, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-12, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-13, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-14, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-15, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-16, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-17, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-18, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-19, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-20, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-21, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-22, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-23, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-24, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-25, or a salt thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-26, or a salt thereof.

[0113] In another aspect, the method comprises administering to a subject a therapeutically effective amount of a combination of any one of the compounds of Formula (I), Formula (II), or compounds I-18 to I-26, or salts thereof. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compound I-20 with one or more other compounds of Formula (I), Formula (II), or compounds I-18 to I-26. In one aspect, the method comprises administering to a subject a therapeutically effective amount of compounds I-20 and I-1, or salts thereof. In another aspect, the neurological disorder is a stress-related disorder (e.g. chronic stress, post-traumatic stress disorder (PTSD), anxiety, or general anxiety disorder). In another aspect, the neurological disorder is an eating disorder (e.g. orthorexia, or anorexia nervosa). In another aspect, the neurological disorder is a pain-related disorder (e.g. chronic pain, primary headache disorder, or migraine). In another aspect, the neurological disorder is a panic disorder (e.g., panic attacks). In another aspect, the neurological disorder is addiction (e.g. addiction to a substance (e.g. addiction to cocaine, amphetamines, methamphetamine, methylphenidate, nicotine, alcohol, prescription medication, marijuana, tobacco, food, or an opioid selected from heroin, fentanyl, codeine, hydrocodone, morphine, oxycodone, hydromorphone, and methadone)).

[0114] In another aspect, the invention provides a method of modulating the activity of a PACAP-receptor (e.g. PAC1) comprising administering an effective amount of any one of the compounds of Formula (I), Formula (II), or compounds I-18 to I-26, or a salt thereof. In another aspect, the modulation is inhibition. In another aspect, the modulation occurs in a biological sample (e.g. tissue or cell). In another aspect, the modulation occurs in a subject. In another aspect, PAC1 receptor antagonism can prevent relapse of an addiction. In certain aspects, PAC1 receptor antagonism can prevent stress-induced relapse of an addiction.

[0115] The invention provides a method of treating a subject suffering from or susceptible to a PACAP receptor-mediated disease or disorder, the method comprising administering to the subject in need thereof a therapeutically effective amount of any one of compounds of Formula (I), Formula (II), or compounds I-18 to I-26, or a salt thereof. In another aspect, the invention provides a method of treating a subject suffering from or susceptible to a PACAP receptor-mediated disease or disorder, the method comprising administering to the subject in need thereof a therapeutically effective amount of compound I-1, or a salt thereof. In another aspect, the PACAP receptor-mediated disorder is a neurological disorder. In another aspect, the PACAP receptor-mediated disease or disorder is a stress-related disorder (e.g. chronic stress, post-traumatic stress disorder (PTSD), anxiety, or general anxiety disorder). In another aspect, the PACAP receptor-mediated disease or disorder is a pain-related disorders (e.g. chronic pain, primary headache disorders, or migraines).

[0116] In certain embodiments, a compound of Formula (I), Formula (II), or compounds I-18 to I-26, is administered in combination with one or more additional pharmaceutical agents described herein. The additional pharmaceutical agent may also be a PACAP receptor antagonist. In certain embodiments, the additional pharmaceutical agent is an additional compound of Formula (I), Formula (II), or compounds I-18 to I-26, or a salt thereof. In certain embodiments, the additional pharmaceutical agent is an inhibitor of a kinase. In certain embodiments, the additional pharmaceutical agent is an inhibitor of MEK/ERK. In certain embodiments, the additional pharmaceutical agent is an anti-inflammatory agent, pain-relieving agent, antihistamine, beta-blocker, anxiolytic, anti-depressant, selective serotonin reuptake inhibitor (SSRI), serotonin and norepinephrine reuptake inhibitor (SNRI), sympatholytic, monoamine oxidase inhibitor (MAOI), opioid, carbamate, benzodiazepine, barbiturate, GABAergic drug, antipsychotic, or other agent.

[0117] The additional pharmaceutical agents include, but are not limited to, anti-anxiety agents, anti-inflammatory agents, antihistamines, anxiolytics, antidepressants, anti-allergic agents, pain-relieving agents, and any combination thereof. In certain embodiments, a pharmaceutical composition described herein further comprises a combination of the additional pharmaceutical agents described herein.

[0118] The inventive compounds or compositions may synergistically modulate a PACAP receptor induced by the additional pharmaceutical agent(s) in the biological sample or subject. Thus, the combination of the inventive compounds or compositions and the additional pharmaceutical agent(s) may be useful in treating neurological diseases resistant to a treatment using the additional pharmaceutical agent(s) without the inventive compounds or compositions.

[0119] In another aspect, the invention provides a kit for treating a neurological disease or disorder comprising a therapeutically effective amount of a compound of Formula (I), Formula (II), or compounds I-18 to I-26, or a salt thereof, and instructions for administration of the compound, or a salt thereof, to a subject. In another aspect, the neurological disease or disorder is a stress-related disorder (e.g. chronic stress, post-traumatic stress disorder (PTSD), anxiety, or general anxiety disorder), eating disorders (e.g. orthorexia, or anorexia nervosa), pain-related disorder (e.g. chronic pain, primary headache disorders, or migraines), panic disorders, panic attacks, or addiction (e.g. addiction to a substance (e.g. addiction to cocaine, amphetamines, methamphetamine, methylphenidate, nicotine, alcohol, prescription medication, marijuana, tobacco, food, or an opioid selected from heroin, fentanyl, codeine, hydrocodone, morphine, oxycodone, hydromorphone, and methadone)).

[0120] In another aspect, the invention provides a method of ameliorating a disorder or symptom thereof in a subject, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of Formula (I), Formula (II), or compounds I-18 to I-26, or a salt thereof. In another aspect, the disease is a neurological disease or disorder.

[0121] Determination of a therapeutically effective amount or a prophylactically effective amount of the compound of the invention, can be readily made by the physician or veterinarian (the “attending clinician”), as one skilled in the art, by the use of known techniques and by observing results obtained under analogous circumstances. The dosages may be varied depending upon the requirements of the patient in the judgment of the attending clinician; the severity of the condition being treated and the particular compound being employed. In determining the therapeutically effective amount or dose, and the prophylactically effective amount or dose, a number of factors are considered by the attending clinician, including, but not limited to: the specific neurological disorder involved; pharmacodynamics characteristics of the particular agent and its mode and route of administration; the desired time course of treatment; the species of mammal; its size, age, and general health; the specific disease involved; the degree of or involvement or the severity of the disease; the response of the individual patient; the particular compound administered; the mode of administration; the bioavailability characteristics of the preparation administered; the dose regimen selected; the kind of concurrent treatment (i.e., the interaction of the compound of the invention with other co-administered therapeutics); and other relevant circumstances.

[0122] Treatment can be initiated with smaller dosages, which are less than the optimum dose of the compound. Thereafter, the dosage may be increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day if desired. A therapeutically effective amount and a prophylactically effective amount of a compound of the invention is expected to vary from about 0.1 milligram per kilogram of body weight per day (mg/kg/day) to about 100 mg/kg/day.

[0123] Compounds determined to be effective for the prevention or treatment of neurological disorders in animals, e.g., dogs, chickens, and rodents, may also be useful in treatment of neurological disorders in humans. Those skilled in the art of treating neurological disorders in humans will know, based upon the data obtained in animal studies, the dosage and route of administration of the compound to humans. In general, the dosage and route of administration in humans is expected to be similar to that in animals.

[0124] The identification of those patients who are in need of prophylactic treatment for neurological disorders is well within the ability and knowledge of one skilled in the art. Certain of the methods for identification of patients which are at risk of developing neurological disorders which can be treated by the subject method are appreciated in the medical arts, such as family history, and the presence of risk factors associated with the development of that disease state in the subject patient. A clinician skilled in the art can readily identify such candidate patients, by the use of, for example, clinical tests, physical examination and medical/family history.

[0125] A method of assessing the efficacy of a treatment in a subject includes determining the pre-treatment extent of a neurological disorder by methods well known in the art and then administering a therapeutically effective amount of a compound of Formula (I), Formula (II), or compounds I-18 to I-26, according to the invention to the subject. After an appropriate period of time after the administration of the compound (e.g., 1 day, 1 week, 2 weeks, one month, six months), the extent of the neurological disorder is determined again. The modulation (e.g., decrease) of the extent of the neurological disorder indicates efficacy of the treatment. The extent of the neurological disorder may be determined periodically throughout treatment. For example, the extent of the neurological disorder may be checked every few hours, days or weeks to assess the further efficacy of the treatment. A decrease in extent of the neurological disorder indicates that the treatment is efficacious. The method described may be used to screen or select patients that may benefit from treatment with a PACAP receptor antagonist for a neurological disorder.

[0126] As used herein, “obtaining a biological sample from a subject,” includes obtaining a sample for use in the methods described herein. A biological sample is described above.

[0127] In another aspect, a compound of the invention or pharmaceutical composition thereof is packaged in a therapeutically effective amount with a pharmaceutically acceptable carrier or diluent. The composition may be formulated for treating a subject suffering from or susceptible to a neurological disorder, and packaged with instructions to treat a subject suffering from or susceptible to a neurological disorder.

[0128] In another aspect, methods of treating a neurological disorder in a subject include administering an effective amount of a compound of the invention to the subject. The administration may be by any route of administering known in the pharmaceutical arts. The subject may have a neurological disorder, may be at risk of developing a neurological disorder, or may need prophylactic treatment prior to anticipated or unanticipated exposure to conditions capable of increasing susceptibility to neurological disorder.

[0129] In one aspect, a method of monitoring the progress of a subject being treated includes determining the pre-treatment status of the neurological disorder, administering a therapeutically effective amount of compound of the invention to the subject, and determining the status of the neurological disorder after an initial period of treatment, wherein the modulation of the status indicates efficacy of the treatment.

[0130] The subject may be at risk of a neurological disorder, may be exhibiting symptoms of a neurological disorder, may be susceptible to a neurological disorder and/or may have been diagnosed with a neurological disorder (e.g. stress-related disorders (e.g. chronic stress, post-traumatic stress disorder (PTSD), anxiety, or general anxiety disorder), eating disorders (e.g. orthorexia, or anorexia nervosa), pain-related disorders (e.g. chronic pain, primary headache disorders, or migraines), panic disorders, panic attacks, or addiction (e.g. addiction to a substance (e.g. addiction to cocaine, amphetamines, methamphetamine, methylphenidate, nicotine, alcohol, prescription medication, marijuana, tobacco, food, or an opioid selected from heroin, fentanyl, codeine, hydrocodone, morphine, oxycodone, hydromorphone, and methadone)).

[0131] The initial period of treatment may be the time in which it takes to establish a stable and/or therapeutically effective blood serum level of any of the compounds, compound combinations, dosing regimens, or pharmaceutical compositions delineated herein, or the time in which it take for the subject to clear a substantial portion of the compound, or any period of time selected by the subject or healthcare professional that is relevant to the treatment.

[0132] If the modulation of the status indicates that the subject may have a favorable clinical response to the treatment, the subject may be treated with the compound. For example, the subject can be administered a therapeutically effective dose or doses of the compound.

[0133] In another aspect, the invention provides antagonists for PACAP-receptors (e.g. PAC1) and methods for their use in a biological sample (e.g. tissue or cell). The methods include contacting the biological sample (e.g. tissue or cell) with an effective amount of any of the compounds of Formula (I), Formula (II), or compounds I-18 to I-26, compound combinations, dosing regimens, or pharmaceutical compositions delineated herein, such that the signaling of the PACAP-receptor is reduced. The contacting may be in vitro, e.g., by addition of the compound to a fluid surrounding the cells, for example, to the growth media in which the cells are living or existing. The contacting may also be by directly contacting the compound to the cells. Alternately, the contacting may be in vivo, e.g., by passage of the compound through a subject; for example, after administration, depending on the route of administration, the compound may travel through the digestive tract or the blood stream or may be applied or administered directly to cells in need of treatment.

[0134] The PACAP-receptor may be within a cell, isolated from a cell, recombinantly expressed, purified or isolated from a cell or recombinant expression system or partially purified or isolated from a cell or recombinant expression system.

[0135] The contacting may be in vitro, e.g., by addition of the compound to a solution containing purified PACAP-receptor, or, if the PACAP-receptor is present in cells, by adding the compound to a fluid surrounding the cells, for example, to the growth media in which the cells are living or existing. The contacting may also be by directly contacting the compound to the cells. Alternately, the contacting may be in vivo, e.g., by passage of the compound through a subject; for example, after administration, depending on the route of administration, the compound may travel through the digestive tract or the blood stream or may be applied or administered directly to cells in need of treatment.

[0136] Kits of the invention include kits for treating a neurological disorder in a subject. The invention also includes kits for regulation of PACAP-receptor function, assessing the efficacy of a treatment for a neurological disorder in a subject, monitoring the progress of a subject being treated for a neurological disorder, selecting a subject with a neurological disorder for treatment according to the invention, and/or treating a subject suffering from or susceptible to a neurological disorder. The kit may include any of the compounds, compound combinations, dosing regimens, or pharmaceutical compositions delineated herein and instructions for use. The instructions for use may include information on dosage, method of delivery, storage of the kit, etc. The kits may also include reagents, for example, test compounds, buffers, media (e.g., cell growth media), cells, etc. Test compounds may include known compounds or newly discovered compounds, for example, combinatorial libraries of compounds. One or more of the kits of the invention may be packaged together, for example, a kit for assessing the efficacy of a treatment for a neurological disorder may be packaged with a kit for monitoring the progress of a subject being treated for a neurological disorder according to the invention.

[0137] The present methods can be performed on cells in culture, e.g. in vitro or ex vivo, or on cells present in an animal subject, e.g., in vivo. Compounds of the inventions can be initially tested in vitro using primary cultures.

[0138] Alternatively, the effects of compound of the invention can be characterized in vivo using animals models. One skilled in the art, provided with the present disclosure, will readily understand that analogous screening methods for the same or a similar library of compounds could now be usefully applied to different neurological conditions in which the same or similar underlying pathologic mechanism is known or suspected to be a factor in the development or manifestation of the disease.

[0139] The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

[0140] The present disclosure provides methods of modulating (e.g., inhibiting or increasing) the activity (e.g., aberrant activity, or undesired activity, such as increased or decreased activity) of a PACAP-receptor. The present disclosure provides methods of modulating (e.g., inhibiting or increasing) the activity (e.g., aberrant activity, such as increased or decreased activity) of a PACAP-receptor in a subject. The present disclosure also provides methods for the treatment of a wide range of diseases, such as diseases associated with the aberrant activity (e.g., increased or decreased activity) of a PACAP-receptor, e.g., neurological diseases or disorders (e.g. stress-related disorders (e.g. chronic stress, post-traumatic stress disorder (PTSD), anxiety, or general anxiety disorder), eating disorders (e.g. orthorexia, or anorexia nervosa), pain-related disorders (e.g. chronic pain, primary headache disorders, or migraines), panic disorders, panic attacks, or addiction (e.g. addiction to a substance (e.g. addiction to cocaine, amphetamines, methamphetamine, methylphenidate, nicotine, alcohol, prescription medication, marijuana, tobacco, food, or an opioid selected from heroin, fentanyl, codeine, hydrocodone, morphine, oxycodone, hydromorphone, and methadone)).

EXAMPLES

[0141] In order that the present disclosure may be more fully understood, the following examples are set forth. The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions, and methods provided herein and are not to be construed in any way as limiting their scope.

Example 1. Compound 1 Blocks Pac1 Receptor Internalization and Calcium Signaling

[0142] Compound I-1 blocks PAC1 receptor internalization and calcium signaling. FIG. 1 shows that PAC1-EGFP receptor is preferentially expressed at the plasma membrane in a stable HEK cell line (Control). Upon 25 nM PACAP application (15 min), the tagged receptor is internalized into intracellular endosomes for ERK signaling (PACAP, middle panel). However, pretreatment of the culture (15 min) with 10-25 μM I-1 blocked PACAP-stimulated receptor internalization (PACAP+I-1); the PAC1-EGFP receptors remained on the cell surface. In FIG. 2, using the same HEK PAC1-EGFP cell line, PACAP rapidly stimulated calcium signaling in Fura-2 assays. The response was blocked approximately 50% with 2.5 μM compound I-1, and was abolished with 25 μM I-1.

Example 2. Compound 1 Blocks PAC1 Receptor-Mediated ERK Activation

[0143] In FIG. 3, using the stably expressing HEK PAC1-EGFP receptor cell line, compound I-1 blocks 25 nM PACAP-stimulated ERK phosphorylation and activation (p42 and p44 ERK bands) in a dose-dependent manner. Compound I-1 at a concentration of 25 μM can block 25 nM PACAP-stimulated ERK activation to near control levels. In FIG. 4, normalized to total ERK levels, I-1 demonstrates an approximate IC.sub.50 of 8-10 μM in inhibiting PACAP-stimulated ERK activation. In similar assays, the other compounds (I-2 to I-26) can block PACAP-stimulated ERK signaling with an apparent IC.sub.50 of 25 μM.

Example 3. Compounds 1 and 10 can Work Synergistically in Blocking ERK Signaling

[0144] In FIG. 5, the compound I-20 acts as a glucagon receptor (GCGR) antagonist by binding to an extra-helical allosteric site on the receptor. I-20 is a weak antagonist at the PAC1-receptor compared to I-1; ERK activation is attenuated approximately 50% with 25 μM I-20. In FIG. 6, the addition of compound I-20 can increase the efficacy and potency of compound I-20 in inhibiting PACAP-stimulated ERK activation. Compared to FIG. 2, compound I-20+10 μM I-1 can block ERK activation to basal or near-basal levels.

Example 4. Comparison of Different Compounds for PAC1 Receptor-Mediated ERK Activation

[0145] In FIG. 7, all samples were pretreated with 25 μM acyl hydrazide before 25 nM PACAP application. Glucagon antagonist is I-20. Data normalized to total cellular ERK not shown.

Example 5. Compound 1 Blocks PACAP-Induced Anxiety-Related Behavior

[0146] Compound I-1 blocks PACAP-induced anxiety-related behavior. In FIG. 8, male rats were cannulated bilaterally into the BNST. In the left panel, in open field tests, 5-10 μM I-1 infusions into the BNST had no apparent effects compared to control. PACAP infusions into the BNST produced stress/anxiety-like responses as reflected by decreased center field times (duration in seconds). However, BNST preinjection with compound I-1 completely blocked the PACAP-induced stress/anxiety responses (I-1+PACAP). In the right panel, the open field entry data was not affected by changes in animal locomotion. Asterisk, statistically different from other groups. Note: I-1+PACAP returned open field times to untreated control levels.

Example 6. Compounds of Formula Blocks PACAP-Stimulated Calcium Signaling

[0147] FIG. 9-17 show the compounds of Formula (I), Formula (II), or compounds I-18 to I-26, demonstrate varying abilities to block PACAP-stimulated calcium signaling. The PAC1-EGFP HEK cell lines were preloaded with Fura-2 for calcium imaging. PACAP alone produced robust calcium signals. At 2.5 μM, most compounds were able to block the PACAP-induced calcium signal approximately 50%; 25 μM compounds blocked calcium flux completely.

Example 7. Characterization Data for Exemplary Compounds

(E)-3-chloro-N′-((1-(3,5-difluorobenzyl)-1H-indol-4-yl)methylene)-4-hydroxybenzohydrazide (I-1)

[0148] ##STR00043##

[0149] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 11.72 (s, 1H), 10.94 (s, 1H), 8.67 (s, 1H), 8.02 (d, J=2.15 Hz, 1H), 7.81 (d, J=8.78 Hz, 1H), 7.69 (d, J=2.54 Hz, 1H), 7.58 (d, J=8.0 Hz, 1H), 7.32 (d, J=7.45 Hz, 1H), 7.29 (d, J=2.17 Hz, 1H), 7.21 (t, J=7.57 Hz, 1H), 7.13, (tt, J=9.47, 2.04 Hz, 1H), 7.10 (d, J=8.45 Hz, 1H), 6.92-6.86 (m, 2H), 5.53 (s, 2H). HRMS (ESI) m/z: [M+H].sup.+ calcd for [C.sub.23H.sub.17ClF.sub.2N.sub.3O.sub.2].sup.+: 440.0977, observed: 440.0973.

(E)-3-chloro-4-hydroxy-N′-((1-(3-nitrobenzyl)-1H-indol-4-yl)methylene)benzohydrazide (I-2)

[0150] ##STR00044##

[0151] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 11.71 (s, 1H), 10.94 (s, 1H), 8.67 (s, 1H), 8.14-8.10 (m, 1H), 8.07 (s, 1H), 8.00 (s, 1H), 7.80 (d, J=7.94 Hz, 1H), 7.72 (s, 1H), 7.64-7.58 (m, 3H), 7.33-7.27 (m, 2H), 7.20 (t, J=7.94 Hz, 1H) 7.08 (d, J=7.94 Hz, 1H), 5.67, (s, 2H). HRMS (ESI) m/z: [M+H].sup.+ calcd for [C.sub.23H.sub.18ClN.sub.4O.sub.4].sup.+: 449.1017, observed: 449.1026.

(E)-3-chloro-N′-((1-(3-cyanobenzyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)methylene)-4-hydroxybenzohydrazide (I-3)

[0152] ##STR00045##

[0153] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 11.99 (s, 1H), 11.01 (s, 1H), 8.67 (s, 1H), 8.35 (d, J=4.47 Hz, 1H), 8.01 (s, 1H), 7.85-7.70 (m, 4H), 7.55-7.50 (m, 2H), 7.35 (s, 1H), 7.13 (s, 1H), 7.10 (d, J=8.67 Hz, 1H), 5.59 (s, 2H).

[0154] .sup.1H NMR (500 MHz, CD.sub.3CN): δ: 10.43 (s, 1H), 8.56 (s, 1H), 8.36 (d, J=5.43 Hz, 1H), 7.99 (d, J=1.43 Hz, 1H), 7.78 (dd, J=8.53, 2.13 Hz, 1H), 7.64 (dt, J=7.63, 1.38 Hz, 1H), 7.61 (s, 1H), 7.55 (s, 1H), 7.53-7.46 (m, 2H), 7.33 (d, J=4.27 Hz, 1H), 7.18 (s, 1H), 7.10 (d, J=8.53 Hz, 1H), 5.59 (s, 2H). HRMS (ESI) m/z: [M+H].sup.+ calcd for [C.sub.23H.sub.17ClN.sub.5O.sub.2].sup.+: 430.1071, observed: 430.1077.

(E)-N′-((1-(3,5-bis(trifluoromethyl)benzyl)-1H-indol-4-yl)methylene)-3-chloro-4-hydroxybenzohydrazide (I-4)

[0155] ##STR00046##

[0156] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 11.72 (s, 1H), 10.95 (s, 1H), 8.66 (s, 1H), 8.03 (s, 1H), 8.02-8.00 (m, 1H), 7.88 (s, 2H), 7.80 (d, J=8.5 Hz, 1H), 7.77 (s, 1H), 7.65 (d, J=8.5 Hz, 1H), 7.34-7.29 (m, 2H), 7.22 (t, J=7.8 Hz, 1H), 7.09 (d, J=7.8 Hz, 1H), 5.72 (s, 2H). HRMS (ESI) m/z: [M+H].sup.+ calcd for [C.sub.25H.sub.17ClF.sub.6N.sub.3O.sub.2].sup.+: 540.0913, observed: 540.0928.

(E)-3-chloro-N′-((1-(4-cyanobenzyl)-1H-indol-4-yl)methylene)-4-hydroxybenzohydrazide (I-5)

[0157] ##STR00047##

[0158] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 11.70 (s, 1H), 10.95 (s, 1H), 8.65 (s, 1H), 8.00 (d, J=1.66 Hz, 1H), 7.79 (app d, J=8.5 Hz, 3H), 7.67 (s, 1H), 7.52 (d, J=8.5 Hz, 1H), 7.34-7.25 (m, 4H), 7.18 (t, J=7.39 Hz, 1H), 7.07 (d, J=8.31 Hz, 1H), 5.61 (s, 2H). HRMS (ESI) m/z: [M+H].sup.+ calcd for [C.sub.24H.sub.18C1N.sub.4O.sub.2].sup.+: 429.1118, observed: 429.1113.

(E)-3-chloro-N′-((1-(3-cyanobenzyl)-1H-indol-4-yl)methylene)-4-hydroxybenzohydrazide (I-6)

[0159] ##STR00048##

[0160] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 11.71 (s, 1H), 10.96 (s, 1H), 8.66 (s, 1H), 8.00 (d, J=2.02 Hz, 1H), 7.80 (d, J=7.69 Hz, 1H), 7.74 (d, J=7.55 Hz, 1H), 7.72-7.68 (m, 2H), 7.59 (d, J=8.12 Hz, 1H), 7.53, (t, J=7.69 Hz, 1H), 7.48 (d, J=8.12 Hz, 1H), 7.31 (d, J=7.27 Hz, 1H), 7.27 (s, 1H), 7.20 (t, J=7.27 Hz, 1H), 7.08 (d, J=8.55 Hz, 1H), 5.56 (s, 2H). HRMS (ESI) m/z: [M+H].sup.+ calcd for [C.sub.24H.sub.18ClN.sub.4O.sub.2].sup.+: 429.1113, observed: 429.1120.

methyl (E)-4-((4-((2-(3-chloro-4-hydroxybenzoyl)hydrazineylidene)methyl)-1H-indol-1-yl)methyl)benzoate (I-7)

[0161] ##STR00049##

[0162] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 11.70 (s, 1H), 10.96 (s, 1H), 8.67 (s, 1H), 8.00 (d, J=1.47 Hz, 1H), 7.90 (d, J=8.09 Hz, 1H), 7.80 (d, J=8.09 Hz, 1H), 7.67 (s, 1H), 7.52 (d, J=8.09 Hz, 1H), 7.33-7.24 (m, 4H), 7.18 (t, J=7.46 Hz, 1H), 7.08 (d, J=8.83 Hz, 1H), 5.60 (s, 2H), 3.82 (s, 3H). HRMS (ESI) m/z: [M+H].sup.+ calcd for [C.sub.25H.sub.21ClN.sub.3O.sub.4].sup.+: 462.1221, observed: 462.1221

(E)-N′-(4-((4-(tert-butyl)benzyl)oxy)-3,5-dimethoxybenzylidene)-3-chloro-4-hydroxybenzohydrazide (I-18)

[0163] ##STR00050##

[0164] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ11.71 (s, 1H), 10.95 (s, 1H), 8.36 (s, 1H), 7.95 (s, 1H), 7.76 (dd, J=8.6, 2.1 Hz, 1H), 7.42-7.34 (m, 4H), 7.07 (d, J=8.6 Hz, 1H), 7.02 (s, 2H), 4.91 (s, 2H), 3.83 (s, 6H), 1.28 (s, 9H). HRMS (ESI) m/z: [M+H].sup.+ calcd for [C.sub.27H.sub.30ClN.sub.2O.sub.5].sup.+: 497.1843, observed: 497.1848.

(E)-3-chloro-4-hydroxy-N′-((1-(2,3,5,6-tetramethylbenzyl)-1H-indol-4-yl)methylene)benzohydrazide (I-19)

[0165] ##STR00051##

[0166] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 11.66 (s, 1H), 10.92 (s, 1H), 8.68 (s, 1H), 7.99 (s, 1H), 7.81-7.73 (m, 2H), 7.35 (d, J=6.98 Hz, 1H), 7.27 (t, J=7.42 Hz, 1H), 7.10-7.03 (m, 3H), 6.79 (s, 1H), 5.38 (s, 2H), 2.23 (s, 6H), 2.10 (s, 6H). HRMS (ESI) m/z: [M+H].sup.+ calcd for [C.sub.27H.sub.27ClN.sub.3O.sub.2].sup.+: 460.1792, observed: 460.1794.

EQUIVALENTS AND SCOPE

[0167] 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 disclosure 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 disclosure 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.

[0168] Furthermore, the disclosure 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 disclosure, or aspects described herein, is/are referred to as comprising particular elements and/or features, certain embodiments described herein or aspects described herein 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 described herein, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0169] 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 disclosure 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 described herein can be excluded from any claim, for any reason, whether or not related to the existence of prior art.

[0170] 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 disclosure, as defined in the following claims.

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Abstract

Claims

Description