OXYSTEROLS AND METHODS OF USE THEREOF

Abstract

Compounds are provided according to Formula (I) and pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof; wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, and R.sup.8 are as defined herein. Compounds of the present invention are contemplated useful for the prevention and treatment of a variety of conditions.

##STR00001##

Claims

1. (canceled)

2. A compound of Formula (I): ##STR00054## or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 is hydrogen or substituted or unsubstituted C.sub.1-6 alkyl; each of R.sup.2 and R.sup.3 is independently hydrogen, substituted or unsubstituted C.sub.1-6 alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, or R.sup.2 and R.sup.3, together with the carbon atom to which they are attached, form a 3-8 membered ring; each of R.sup.4 and R.sup.5 is independently hydrogen or a moiety cleavable under biological conditions; R.sup.8 is absent or hydrogen; and represents a single or double bond, wherein when one is a double bond, the other is a single bond and R.sup.8 is absent.

3. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein R.sup.4 and R.sup.5 are not both hydrogen.

4.-5. (canceled)

6. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein each of R.sup.4 and R.sup.5 is independently hydrogen, —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)OR.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or substituted or unsubstituted alkyl; each R.sup.c is independently substituted or unsubstituted alkyl; each R.sup.d is independently hydrogen or substituted or unsubstituted alkyl; each x is independently 1 or 2; and each of n, m, p is independently 1, 2, 3, or 4.

7. (canceled)

8. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein R.sup.1 is substituted or unsubstituted C.sub.1-6 alkyl.

9. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein R.sup.1 is hydrogen.

10.-11. (canceled)

12. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein each of R.sup.2 and R.sup.3 is independently hydrogen, methyl, —CF.sub.3, ethyl, isopropyl, cyclopropyl, or butyl.

13. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein R.sup.4 is a moiety cleavable under biological conditions and R.sup.5 is hydrogen.

14. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein R.sup.4 is hydrogen and R.sup.5 is a moiety cleavable under biological conditions.

15. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein each of R.sup.4 and R.sup.5 is a moiety cleavable under biological conditions.

16. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein: each of R.sup.4 and R.sup.5 is independently hydrogen, —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)OR.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or substituted or unsubstituted alkyl; each R.sup.c is independently substituted or unsubstituted alkyl; each R.sup.d is independently hydrogen or substituted or unsubstituted alkyl; each x is independently 1 or 2; and each of n, m, p is independently 1, 2, 3, or 4.

17. The compound or pharmaceutically acceptable salt thereof, of claim 16, wherein when R.sup.4 is hydrogen and R.sup.5 is —S(O).sub.xR.sup.b and x is 2, R.sup.b is not —OH.

18. The compound or pharmaceutically acceptable salt thereof, of claim 16, wherein not both of R.sup.4 or R.sup.5 are hydrogen.

19. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein R.sup.4 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)OR.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; R.sup.5 is hydrogen; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or substituted or unsubstituted alkyl; each R.sup.c is independently substituted or unsubstituted alkyl; each R.sup.d is independently hydrogen or substituted or unsubstituted alkyl; each x is independently 1 or 2; and each of n, m, p is independently 1, 2, 3, or 4.

20. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein R.sup.4 is hydrogen; R.sup.5 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)OR.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or substituted or unsubstituted alkyl; each R.sup.c is independently substituted or unsubstituted alkyl; each R.sup.d is independently hydrogen or substituted or unsubstituted alkyl; each x is independently 1 or 2; each of n, m, p is independently 1, 2, 3, or 4; wherein when R.sup.5 is —S(O).sub.xR.sup.b and x is 2, R.sup.b is not —OH.

21. The compound or pharmaceutically acceptable salt thereof, of claim 18, wherein R.sup.4 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, or —(CH.sub.2).sub.pOC(O)R.sup.c.

22. The compound or pharmaceutically acceptable salt thereof, of claim 21, wherein each of R.sup.a and R.sup.b is independently —OR.sup.d, R.sup.d is hydrogen or substituted or unsubstituted alkyl, and x is 2.

23. The compound of claim 21, wherein R.sup.c is substituted or unsubstituted alkyl; and R.sup.d is hydrogen or substituted or unsubstituted alkyl.

24. The compound or pharmaceutically acceptable salt thereof, of claim 21, wherein R.sup.c is —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2CO.sub.2H, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2.

25. The compound or pharmaceutically acceptable salt thereof, of claim 21, wherein each of n, m, and p is independently 1 or 2.

26. The compound or pharmaceutically acceptable salt thereof, of claim 21, wherein R.sup.4 is hydrogen, —P(O).sub.2OH, —S(O).sub.2OH, —CH.sub.2OP(O)(OH).sub.2, —C(O)CH.sub.3, —C(O)CH.sub.2NH.sub.2, —C(O)CH.sub.2CH.sub.2C(O)OH, —C(O)CH(CH(CH.sub.3).sub.2)NH.sub.2, —C(O)CH(CH.sub.3)NH.sub.2, or any amino acid residue.

27. The compound or pharmaceutically acceptable salt thereof, of claim 16, wherein R.sup.5 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, or —(CH.sub.2).sub.pOC(O)R.sup.c.

28. The compound or pharmaceutically acceptable salt thereof, of claim 27, wherein each of R.sup.a and R.sup.b is independently —OR.sup.d, R.sup.d is hydrogen or substituted or unsubstituted alkyl, and x is 2.

29. The compound or pharmaceutically acceptable salt thereof, of claim 27, wherein R.sup.c is substituted or unsubstituted alkyl; and R.sup.d is hydrogen or substituted or unsubstituted alkyl.

30. The compound or pharmaceutically acceptable salt thereof, of claim 27, wherein each of n, m, and p is independently 1 or 2.

31. The compound or pharmaceutically acceptable salt thereof, of claim 27, wherein R.sup.5 is hydrogen, —P(O).sub.2OH, —S(O).sub.2OH, —CH.sub.2OP(O)(OH).sub.2, —C(O)CH.sub.3, —C(O)CH.sub.2NH.sub.2, —C(O)CH.sub.2CH.sub.2C(O)OH, —C(O)CH(CH(CH.sub.3).sub.2)NH.sub.2, —C(O)CH(CH.sub.3)NH.sub.2, or any amino acid residue.

32. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein when R.sup.4 is hydrogen, R.sup.5 is not —S(O).sub.2OH.

33. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein each of is a singe bond.

34. The compound or pharmaceutically acceptable salt thereof, of claim 2, wherein the compound of Formula (I) is a compound of Formula (I-A) or Formula (I-B): ##STR00055## or a pharmaceutically acceptable salt thereof.

35-63. (canceled)

64. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of: ##STR00056## ##STR00057## ##STR00058## ##STR00059## ##STR00060##

65. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of: ##STR00061##

66. A pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof according to claim 2, and a pharmaceutically acceptable carrier.

67. A method of inducing sedation or anesthesia comprising administering to a subject an effective amount of a compound or a pharmaceutically acceptable salt thereof according to claim 2, or pharmaceutical composition thereof.

68. A method for treating or preventing a disease or disorder, comprising administering to a subject in need thereof an effective amount of a compound or a pharmaceutically acceptable salt thereof according to claim 2, or pharmaceutical composition thereof, wherein the disease or disorder is selected from the group consisting of a gastrointestinal (GI) disorder, inflammatory bowel disease (IBD), a structural disorder affecting the GL an anal disorder, a colon polyp, cancer, diabetes, a sterol synthesis disorder, and colitis.

69-71. (canceled)

72. A method for treating or preventing a CNS-related condition comprising administering to a subject in need thereof an effective amount of a compound or pharmaceutically acceptable salt thereof according to claim 2, or pharmaceutical composition thereof, wherein the CNS-related condition is an adjustment disorder, an anxiety disorder, a cognitive disorder, a dissociative disorder, an eating disorder, a mood disorder, schizophrenia or other psychotic disorder, a sleep disorder, a substance-related disorder, a personality disorder, an autism spectrum disorder, a neurodevelopmental disorder, multiple sclerosis, a sterol synthesis disorder, pain, encephalopathy secondary to a medical condition, a seizure disorder, stroke, traumatic brain injury, a movement disorder, vision impairment, hearing loss, and tinnitus.

73-74. (canceled)

Description

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION

[0113] [1] As generally described herein, the present invention provides oxysterols useful for preventing and/or treating a broad range of disorders, including, but not limited to, NMDA-mediated disorders. These compounds are expected to show improved in vivo potency, pharmacokinetic (PK) properties, oral bioavailability, formulatability, stability, and/or safety as compared to other oxysterols.
[2] The compounds described herein comprise a moiety cleavable under biological conditions, for example by enzymes (e.g., hydrolases).

Compounds

[0114] In one aspect, provided herein are compounds according to Formula (I):

##STR00016##

or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 is hydrogen or C.sub.1-6 alkyl; each of R.sup.2 and R.sup.3 is independently hydrogen, C.sub.1-6 alkyl, or carbocyclyl, or R.sup.2 and R.sup.3, together with the carbon atom to which they are attached, form a 3-8 membered ring; each of R.sup.4 and R.sup.5 is independently hydrogen; R.sup.8 is absent or hydrogen; represents a single or double bond, wherein when one is a double bond, the other is a single bond and R.sup.8 is absent; and at least one hydrogen is replaced by a moiety cleavable under biological conditions.

[0115] In some embodiments, the compound of Formula (I) is a compound of Formula (I-AA):

##STR00017##

or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 is hydrogen or C.sub.1-6 alkyl; each of R.sup.2 and R.sup.3 is independently hydrogen. C.sub.1-6 alkyl, or carbocyclyl, or R.sup.2 and R.sup.3, together with the carbon atom to which they are attached, form a 3-8 membered ring; each of R.sup.4 and R.sup.5 is independently hydrogen or a moiety cleavable under biological conditions; R.sup.8 is absent or hydrogen; and represents a single or double bond, wherein when one is a double bond, the other is a single bond and R.sup.8 is absent.

[0116] In some embodiments, R.sup.4 and R.sup.5 are not both hydrogen.

[0117] In some embodiments, R.sup.4 is not hydrogen.

[0118] In some embodiments, R.sup.5 is not hydrogen.

[0119] In some embodiments, each of R.sup.4 and R.sup.5 is independently hydrogen, —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —C(O)OR.sup.c, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or alkyl; each R.sup.c is independently alkyl (e.g., —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2CO.sub.2H, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2);

each R.sup.d is independently hydrogen or alkyl; each x is independently 1 or 2; and each of n, m, p is independently 1, 2, 3, or 4. In some embodiments, each of R.sup.4 and R.sup.5 is independently —C(O)R.sup.c, wherein R.sup.c is an amino acid (e.g., glycine, alanine, valine). In some embodiments, each of R.sup.4 and R.sup.5 is independently an amino acid ester.

[0120] In some embodiments, R.sup.4 is hydrogen, —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —C(O)OR.sup.c, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or alkyl; each R.sup.c is independently alkyl (e.g., —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2CO.sub.2H, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2);

each R.sup.d is independently hydrogen or alkyl; each x is independently 1 or 2; and each of n, m, p is independently 1, 2, 3, or 4.

[0121] In some embodiments, R.sup.1 is C.sub.1-6 alkyl (e.g., substituted or unsubstituted C.sub.1-6 alkyl). In some embodiments, R.sup.1 is hydrogen. In some embodiments, R.sup.1 is hydrogen, methyl (e.g., —CH.sub.3, —CF.sub.3, —CH.sub.2OCH.sub.3), ethyl, or isopropyl. In some embodiments, R.sup.1 is methyl or ethyl.

[0122] In some embodiments, each of R.sup.2 and R.sup.3 is independently hydrogen, methyl (e.g., —CH.sub.3, —CF.sub.3), ethyl, isopropyl, cyclopropyl, or butyl.

[0123] In some embodiments, R.sup.4 is a moiety cleavable under biological conditions and R.sup.5 is hydrogen.

[0124] In some embodiments, R.sup.4 is hydrogen and R.sup.5 is a moiety cleavable under biological conditions. In some embodiments, each of R.sup.4 and R.sup.5 is a moiety cleavable under biological conditions. In some embodiments, each of R.sup.4 and R.sup.5 is independently hydrogen, —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —C(O)OR.sup.c, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or alkyl; each R.sup.c is independently alkyl (e.g., —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2CO.sub.2H, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2);

each R.sup.d is independently hydrogen or alkyl; each x is independently 1 or 2; and
each of n, m, p is independently 1, 2, 3, or 4.

[0125] In some embodiments, when R.sup.4 is hydrogen and R.sup.5 is —S(O).sub.xR.sup.b and x is 2, R.sup.b is not —OH. In some embodiments, not both of R.sup.4 or R.sup.5 are hydrogen.

[0126] In some embodiments, R.sup.4 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —C(O)OR.sup.c, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; R.sup.5 is hydrogen; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or alkyl; each R.sup.c is independently alkyl (e.g., —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2CO.sub.2H, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2); each R.sup.d is independently hydrogen or alkyl; each x is independently 1 or 2; and each of n, m, p is independently 1, 2, 3, or 4. In some embodiments, R.sup.4 is hydrogen; R.sup.5 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —C(O)OR.sup.c, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or alkyl; each R.sup.c is independently alkyl (e.g., —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2CO.sub.2H, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2); each R.sup.d is independently hydrogen or alkyl; each x is independently 1 or 2; each of n, m, p is independently 1, 2, 3, or 4; wherein when R.sup.5 is —S(O).sub.xR.sup.b and x is 2, R.sup.b is not —OH. In some embodiments, R.sup.4 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, or —(CH.sub.2).sub.pOC(O)R.sup.c. In some embodiments, each of R.sup.a and R.sup.b is independently —OR.sup.d, R.sup.d is hydrogen or alkyl, and x is 2. In some embodiments, R.sup.c is alkyl (e.g., —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2COH, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2); and R.sup.d is hydrogen or alkyl (e.g., methyl).

[0127] In some embodiments, each of n, m, and p is independently 1 or 2.

[0128] In some embodiments, R.sup.4 is hydrogen, —P(O).sub.2OH, —S(O).sub.2OH, —CH.sub.2OP(O)(OH).sub.2, —C(O)CH.sub.3, —C(O)CH.sub.2NH.sub.2, —C(O)CH.sub.2CH.sub.2C(O)OH, —C(O)CH(CH(CH.sub.3).sub.2)NH.sub.2, —C(O)CH(CH.sub.3)NH.sub.2, or any amino acid residue.

[0129] In some embodiments, R.sup.5 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, or —(CH.sub.2).sub.pOC(O)R.sup.c. In some embodiments, each of R.sup.a and R.sup.b is independently —OR.sup.d, R.sup.d is hydrogen or alkyl, and x is 2. In some embodiments, R.sup.c is alkyl (e.g., —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2CO.sub.2H, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2); and R.sup.d is hydrogen or alkyl (e.g., methyl). In some embodiments, each of n, m, and p is independently 1 or 2. In some embodiments, R.sup.5 is hydrogen, —P(O).sub.2OH, —S(O).sub.2OH, —CH.sub.2OP(O)(OH).sub.2, —C(O)CH.sub.3, —C(O)CH.sub.2NH.sub.2, —C(O)CH.sub.2CH.sub.2C(O)OH, —C(O)CH(CH(CH.sub.3).sub.2)NH.sub.2, —C(O)CH(CH.sub.3)NH.sub.2, or any amino acid residue.

[0130] In some embodiments, when R.sup.4 is hydrogen, R.sup.5 is not —S(O).sub.2OH.

[0131] In some embodiments, each of is single bond.

[0132] In some embodiments, the compound of Formula (I) is a compound of Formula (I-A) or Formula (I-B):

##STR00018##

or a pharmaceutically acceptable salt thereof.

[0133] In some embodiments, the compound of Formula (I) is a compound of Formula (I-B):

##STR00019##

or a pharmaceutically acceptable salt thereof.

[0134] In some embodiments, R.sup.1 is alkyl (e.g., substituted or unsubstituted alkyl). In some embodiments, R.sup.1 is hydrogen. In some embodiments, R.sup.1 is hydrogen, methyl (e.g., —CH.sub.3, —CF.sub.3, —CH.sub.2OCH.sub.3), ethyl, or isopropyl.

[0135] In some embodiments, each of R.sup.2 and R.sup.3 is independently hydrogen, methyl (e.g., —CH.sub.3, —CF.sub.3), ethyl, isopropyl, cyclopropyl, or butyl.

[0136] In some embodiments, R.sup.4 is a moiety cleavable under biological conditions and R.sup.5 is hydrogen. In some embodiments, R.sup.4 is hydrogen and R.sup.5 is a moiety cleavable under biological conditions.

[0137] In some embodiments, each of R.sup.4 and R.sup.5 is a moiety cleavable under biological conditions. In some embodiments, each of R.sup.4 and R.sup.5 is independently hydrogen, —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —C(O)OR.sup.c, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or alkyl; each R.sup.c is independently alkyl (e.g., unsubstituted alkyl or substituted alkyl, e.g., —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2CO.sub.2H, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2); each R.sup.d is independently hydrogen or alkyl; each x is independently 1 or 2; and each of n, m, p is independently 1, 2, 3, or 4, wherein when R.sup.4 is hydrogen and R.sup.5 is —S(O).sub.x.sup.b and x is 2. R.sup.b is not —OH.

[0138] In some embodiments, not both of R.sup.4 and R.sup.5 are hydrogen.

[0139] In some embodiments, R.sup.4 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —C(O)OR.sup.c, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; R.sup.5 is hydrogen; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or alkyl; each R.sup.c is independently alkyl (e.g., —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2CO.sub.2H, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2); each R.sup.d is independently hydrogen or alkyl; each x is independently 1 or 2; and each of n, m, p is independently 1, 2, 3, or 4.

[0140] In some embodiments, R.sup.4 is hydrogen; R.sup.5 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —C(O)OR.sup.c, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b—(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c; each of R.sup.a and R.sup.b is independently selected from —OR.sup.d or alkyl; each R.sup.c is independently alkyl (e.g., —CH.sub.2NH.sub.2, —CH.sub.2CH.sub.2CO.sub.2H, —CH(CH(CH.sub.3).sub.2)NH.sub.2, —CH.sub.2CH.sub.2C(O)OH, or —CH(CH.sub.3)NH.sub.2); each R.sup.d is independently hydrogen or alkyl; each x is independently 1 or 2; each of n, m, p is independently 1, 2, 3, or 4; wherein when R.sup.5 is —S(O).sub.xR.sup.b and x is 2, R.sup.b is not —OH. In some embodiments, R.sup.4 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, or —(CH.sub.2).sub.pOC(O)R.sup.c. In some embodiments, R.sup.4 is hydrogen, —P(O).sub.2OH, —S(O).sub.2OH, —CH.sub.2OP(O).sub.p(OH).sub.2, —C(O)CH.sub.3, —C(O)CH.sub.2NH.sub.2, —C(O)CH.sub.2CH.sub.2C(O)OH, —C(O)CH(CH(CH.sub.3).sub.2)NH.sub.2, —C(O)CH(CH.sub.3)NH.sub.2, or any amino acid residue.

[0141] In some embodiments, R.sup.5 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, or —(CH.sub.2).sub.pOC(O)R.sup.c. In some embodiments, R.sup.5 is hydrogen, —P(O).sub.2OH, —S(O).sub.2OH, —CH.sub.2OP(O)(OH).sub.2, —C(O)CH.sub.3, —C(O)CH.sub.2NH.sub.2, —C(O)CH.sub.2CH.sub.2C(O)OH, —C(O)CH(CH(CH.sub.3).sub.2)NH.sub.2, —C(O)CH(CH.sub.3)NH.sub.2, or any amino acid residue.

[0142] In some embodiments, when R.sup.4 is hydrogen, R.sup.5 is not —S(O).sub.2OH.

[0143] In some embodiments, the compound of Formula (I-B) is a compound of Formula (I-C):

##STR00020##

or a pharmaceutically acceptable salt thereof.

[0144] In some embodiments, R.sup.1 is hydrogen, methyl (e.g., —CH.sub.3, —CF.sub.3, —CH.sub.2OCH.sub.3), ethyl, or isopropyl. In some embodiments, R.sup.1 is methyl or ethyl.

[0145] In some embodiments, R.sup.4 is a moiety cleavable under biological conditions and R.sup.5 is hydrogen. In some embodiments, R.sup.4 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, or —(CH.sub.2).sub.pOC(O)R.sup.c. In some embodiments, R.sup.4 is hydrogen, —P(O).sub.2OH, —S(O).sub.2OH, —CH.sub.2OP(O)(OH).sub.2, —C(O)CH.sub.3, —C(O)CH.sub.2NH.sub.2, —C(O)CH.sub.2CH.sub.2C(O)OH, —C(O)CH(CH(CH.sub.3).sub.2)NH.sub.2, —C(O)CH(CH.sub.3)NH.sub.2, or any amino acid residue. In some embodiments, R.sup.4 is hydrogen and R.sup.5 is a moiety cleavable under biological conditions.

[0146] In some embodiments, R.sup.5 is —P(O)(R.sup.a).sub.2, —S(O).sub.xR.sup.b, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, or —(CH.sub.2).sub.pOC(O)R.sup.c. In some embodiments, R.sup.5 is hydrogen, —P(O).sub.2OH, —S(O).sub.2OH, —CH.sub.2OP(O)(OH).sub.2, —C(O)CH.sub.3, —C(O)CH.sub.2NH.sub.2, —C(O)CH.sub.2CH.sub.2C(O)OH, —C(O)CH(CH(CH.sub.3).sub.2)NH.sub.2, —C(O)CH(CH.sub.3)NH.sub.2, or any amino acid residue.

[0147] In some embodiments, each of R.sup.4 and R.sup.5 is a moiety cleavable under biological conditions.

[0148] In some embodiments, R.sup.1 is alkyl (e.g., substituted or unsubstituted alkyl).

[0149] In some embodiments, R.sup.1 is hydrogen, and when R.sup.4 is hydrogen, then R.sup.5 is not —S(O).sub.2OH. In some embodiments, R.sup.1 is hydrogen, and when R.sup.4 is hydrogen, then R.sup.5 is —P(O)(R.sup.a).sub.2, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d).sub.2, —C(O)OR.sup.c, —(CH.sub.2).sub.nOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c. In some embodiments, R.sup.1 and R.sup.5 are hydrogen. In some embodiments, R.sup.1 is hydrogen and R.sup.5 is not —S(O).sub.2OH. In some embodiments, R.sup.1 is hydrogen and R.sup.5 is —P(O)(R.sup.a).sub.2, —C(O)R.sup.c, —C(O)N(R.sup.d).sub.2, —(CH.sub.2).sub.xC(O)N(R.sup.d), —C(O)OR.sup.c, —(CH.sub.2).sub.uOP(O)(R.sup.a).sub.2, —(CH.sub.2).sub.mOS(O).sub.xR.sup.b, —(CH.sub.2).sub.pOC(O)R.sup.c, or —(CH.sub.2).sub.pC(O)OR.sup.c.

[0150] In some embodiments, the compound of Formula (I-C) is a compound of Formula (I-D):

##STR00021##

or a pharmaceutically acceptable salt thereof.

[0151] In some embodiments, the compound is selected from the group consisting of:

##STR00022## ##STR00023## ##STR00024## ##STR00025## ##STR00026##

or a pharmaceutically acceptable salt thereof.

[0152] In some embodiments, the compound is selected from the group consisting of:

##STR00027##

Pharmaceutical Compositions

[0153] In another aspect, the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a effective amount of a compound of Formula (I).

[0154] When employed as pharmaceuticals, the compounds provided herein are typically administered in the form of a pharmaceutical composition. Such compositions can be prepared in a manner well known in the pharmaceutical art and comprise at least one active compound.

[0155] In one embodiment, with respect to the pharmaceutical composition, the carrier is a parenteral carrier, oral or topical carrier.

[0156] The present invention also relates to a compound of Formula (I) or pharmaceutical composition thereof for use as a pharmaceutical or a medicament.

[0157] Generally, the compounds provided herein are administered in a therapeutically effective amount. The amount of the compound actually administered will typically be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.

[0158] The pharmaceutical compositions provided herein can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal. Depending on the intended route of delivery, the compounds provided herein are preferably formulated as either injectable or oral compositions or as salves, as lotions or as patches all for transdermal administration.

[0159] The compositions for oral administration can take the form of bulk liquid solutions or suspensions, or bulk powders. More commonly, however, the compositions are presented in unit dosage forms to facilitate accurate dosing. The term “unit dosage forms” refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, premeasured ampules or syringes of the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions. In such compositions, the compound is usually a minor component (from about 0.1 to about 50% by weight or preferably from about 1 to about 40% by weight) with the remainder being various vehicles or carriers and processing aids helpful for forming the desired dosing form.

[0160] Liquid forms suitable for oral administration may include a suitable aqueous or nonaqueous vehicle with buffers, suspending and dispensing agents, colorants, flavors and the like. Solid forms may include, for example, any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

[0161] Injectable compositions are typically based upon injectable sterile saline or phosphate-buffered saline or other injectable carriers known in the art. As before, the active compound in such compositions is typically a minor component, often being from about 0.05 to 10% by weight with the remainder being the injectable carrier and the like.

[0162] Transdermal compositions are typically formulated as a topical ointment or cream containing the active ingredient(s), generally in an amount ranging from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably from about 0.1 to about 10% by weight, and more preferably from about 0.5 to about 15% by weight. When formulated as a ointment, the active ingredients will typically be combined with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients may be formulated in a cream with, for example an oil-in-water cream base. Such transdermal formulations are well-known in the art and generally include additional ingredients to enhance the dermal penetration of stability of the active ingredients or the formulation. All such known transdermal formulations and ingredients are included within the scope provided herein.

[0163] The compounds provided herein can also be administered by a transdermal device. Accordingly, transdermal administration can be accomplished using a patch either of the reservoir or porous membrane type, or of a solid matrix variety.

[0164] The above-described components for orally administrable, injectable or topically administrable compositions are merely representative. Other materials as well as processing techniques and the like are set forth in Part 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pa., which is incorporated herein by reference.

[0165] The above-described components for orally administrable, injectable, or topically administrable compositions are merely representative. Other materials as well as processing techniques and the like are set forth in Part 8 of Remington's The Science and Practice of Pharmacy, 21st edition, 2005, Publisher: Lippincott Williams & Wilkins, which is incorporated herein by reference.

[0166] The compounds of this invention can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.

[0167] The present invention also relates to the pharmaceutically acceptable formulations of a compound of Formula (I). In one embodiment, the formulation comprises water. In another embodiment, the formulation comprises a cyclodextrin derivative. The most common cyclodextrins are α-, β- and γ-cyclodextrins consisting of 6, 7 and 8α-1,4-linked glucose units, respectively, optionally comprising one or more substituents on the linked sugar moieties, which include, but are not limited to, methylated, hydroxyalkylated, acylated, and sulfoalkylether substitution. In certain embodiments, the cyclodextrin is a sulfoalkyl ether β-cyclodextrin, e.g., for example, sulfobutyl ether β-cyclodextrin, also known as Captisol®. Sec. e.g., U.S. Pat. No. 5,376,645. In certain embodiments, the formulation comprises hexapropyl-β-cyclodextrin. In a more particular embodiment, the formulation comprises hexapropyl-β-cyclodextrin (10-50% in water).

[0168] The present invention also relates to the pharmaceutically acceptable acid addition salt of a compound of Formula (I). The acid which may be used to prepare the pharmaceutically acceptable salt is that which forms a non-toxic acid addition salt, i.e., a salt containing pharmacologically acceptable anions such as the hydrochloride, hydroiodide, hydrobromide, nitrate, sulfate, bisulfate, phosphate, acetate, lactate, citrate, tartrate, succinate, maleate, fumarate, benzoate, para-toluenesulfonate, and the like.

[0169] The following formulation examples illustrate representative pharmaceutical compositions that may be prepared in accordance with this invention. The present invention, however, is not limited to the following pharmaceutical compositions.

[0170] Exemplary Formulation 1—Tablets: A compound of Formula (I), or pharmaceutically acceptable salt thereof, may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 240-270 mg tablets (80-90 mg of active compound per tablet) in a tablet press.

[0171] Exemplary Formulation 2—Capsules: A compound of Formula (I), or pharmaceutically acceptable salt thereof, may be admixed as a dry powder with a starch diluent in an approximate 1:1 weight ratio. The mixture is filled into 250 mg capsules (125 mg of active compound per capsule).

[0172] Exemplary Formulation 3—Liquid: A compound of Formula (I), or pharmaceutically acceptable salt thereof, (125 mg) may be admixed with sucrose (1.75 g) and xanthan gum (4 mg) and the resultant mixture may be blended, passed through a No. 10 mesh U.S. sieve, and then mixed with a previously made solution of microcrystalline cellulose and sodium carboxymethyl cellulose (11:89, 50 mg) in water. Sodium benzoate (10 mg), flavor, and color are diluted with water and added with stirring. Sufficient water may then be added to produce a total volume of 5 mL.

[0173] Exemplary Formulation 4—Tablets: A compound of Formula (I), or pharmaceutically acceptable salt thereof, may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 450-900 mg tablets (150-300 mg of active compound) in a tablet press.

[0174] Exemplary Formulation 5—Injection: A compound of Formula (I), or pharmaceutically acceptable salt thereof, may be dissolved or suspended in a buffered sterile saline injectable aqueous medium to a concentration of approximately 5 mg/mL.

[0175] Exemplary Formulation 6—Tablets: A compound of Formula (I), or pharmaceutically acceptable salt thereof, may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 90-150 mg tablets (30-50 mg of active compound per tablet) in a tablet press.

[0176] Exemplary Formulation 7—Tablets: A compound of Formula (I), or pharmaceutically acceptable salt thereof, may be may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 30-90 mg tablets (10-30 mg of active compound per tablet) in a tablet press.

[0177] Exemplary Formulation 8—Tablets: A compound of Formula (I), or pharmaceutically acceptable salt thereof, may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 0.3-30 mg tablets (0.1-10 mg of active compound per tablet) in a tablet press.

[0178] Exemplary Formulation 9—Tablets: A compound of Formula (I), or pharmaceutically acceptable salt thereof, may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 150-240 mg tablets (50-80 mg of active compound per tablet) in a tablet press.

[0179] Exemplary Formulation 10—Tablets: A compound of Formula (I), or pharmaceutically acceptable salt thereof, may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate is added as a lubricant. The mixture is formed into 270-450 mg tablets (90-150 mg of active compound per tablet) in a tablet press.

[0180] Injection dose levels range from about 0.1 mg/kg/hour to at least 10 mg/kg/hour, all for from about 1 to about 120 hours and especially 24 to 96 hours. A preloading bolus of from about 0.1 mg/kg to about 10 mg/kg or more may also be administered to achieve adequate steady state levels. The maximum total dose is not expected to exceed about 2 g/day for a 40 to 80 kg human patient.

[0181] For the prevention and/or treatment of long-term conditions the regimen for treatment usually stretches over many months or years so oral dosing is preferred for patient convenience and tolerance. With oral dosing, one to five and especially two to four and typically three oral doses per day are representative regimens. Using these dosing patterns, each dose provides from about 0.01 to about 20 mg/kg of the compound provided herein, with preferred doses each providing from about 0.1 to about 10 mg/kg, and especially about 1 to about 5 mg/kg.

[0182] Transdermal doses are generally selected to provide similar or lower blood levels than are achieved using injection doses.

[0183] When used to prevent the onset of a CNS-disorder, the compounds provided herein will be administered to a subject at risk for developing the condition, typically on the advice and under the supervision of a physician, at the dosage levels described above. Subjects at risk for developing a particular condition generally include those that have a family history of the condition, or those who have been identified by genetic testing or screening to be particularly susceptible to developing the condition.

Methods of Treatment and Use

[0184] Compounds of the present invention, e.g., a compound of Formula (I), and pharmaceutically acceptable salts thereof, as described herein, are generally designed to modulate NMDA function, and therefore to act as oxysterols for the treatment and prevention of, e.g., CNS-related conditions in a subject. In some embodiments, the compounds described herein, e.g., a compound of Formula (I), and pharmaceutically acceptable salts thereof, as described herein, are generally designed to penetrate the blood brain barrier (e.g., designed to be transported across the blood brain barrier). Modulation, as used herein, refers to, for example, the inhibition or potentiation of NMDA receptor function. In certain embodiments, the compound of Formula (D, or pharmaceutically acceptable salt thereof, may act as a negative allosteric modulator (NAM) of NMDA, and inhibit NMDA receptor function. In certain embodiments, the present invention, e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof, may act as positive allosteric modulators (PAM) of NMDA, and potentiate NMDA receptor function. In certain embodiments, the compound of Formula (I), or pharmaceutically acceptable salt thereof, modulates NMDA function, but does not act as a negative allosteric modulator (NAM) or positive allosteric modulator (PAM) of NMDA.

[0185] In some embodiments, the disorder is cancer. In some embodiments, the disorder is diabetes. In some embodiments, the disorder is a sterol synthesis disorder. In some embodiments, the disorder is a gastrointestinal (GI) disorder. e.g., constipation, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD) (e.g., ulcerative colitis. Crohn's disease), structural disorders affecting the GI, anal disorders (e.g., hemorrhoids, internal hemorrhoids, external hemorrhoids, anal fissures, perianal abscesses, anal fistula), colon polyps, cancer, colitis. In some embodiments, the disorder is inflammatory bowel disease.

[0186] In some embodiments, the disorder is Smith-Lemli-Opitz Syndrome (SLOS). In some embodiments, the disorder is desmosterolosis. In some embodiments, the disorder is sitosterolemia. In some embodiments, the disorder is cerebrotendinous xanthomatosis (CTX). In some embodiments, the disorder is Mevalonate Kinase Deficiency (MKD). In some embodiments, the disorder is SC4MOL gene mutation (SMO Deficiency). In some embodiments, the disorder is Niemann-Pick disease. In some embodiments, the disorder is autism spectrum disorder (ASD). In some embodiments, the disorder is associated with phenylketomuria.

[0187] Exemplary conditions related to NMDA-modulation includes, but are not limited to, gastrointestinal (GI) disorder, e.g., constipation, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD) (e.g., ulcerative colitis. Crohn's disease), structural disorders affecting the GI, anal disorders (e.g., hemorrhoids, internal hemorrhoids, external hemorrhoids, anal fissures, perianal abscesses, anal fistula), colon polyps, cancer, colitis, and CNS conditions. e.g., as described herein.

[0188] Exemplary CNS conditions related to NMDA-modulation include, but are not limited to, adjustment disorders, anxiety disorders (including obsessive-compulsive disorder, posttraumatic stress disorder, social phobia, generalized anxiety disorder), cognitive disorders (including Alzheimer's disease and other forms of dementia (e.g., frontotemporal dementia)), dissociative disorders, eating disorders, mood disorders (including depression (e.g., postpartum depression), bipolar disorder, dysthymic disorder, suicidality), schizophrenia or other psychotic disorders (including schizoaffective disorder), sleep disorders (including insomnia), substance abuse-related disorders, personality disorders (including obsessive-compulsive personality disorder), autism spectrum disorders (including those involving mutations to the Shank group of proteins (e.g., Shank3)), neurodevelopmental disorders (including Rett syndrome), multiple sclerosis, sterol synthesis disorders, pain (including acute and chronic pain; headaches. e.g., migraine headaches), seizure disorders (including status epilepticus and monogenic forms of epilepsy such as Dravet's disease, and Tuberous Sclerosis Complex (TSC)), stroke, traumatic brain injury, movement disorders (including Huntington's disease and Parkinson's disease) and tinnitus. In certain embodiments, the compound of the present invention. e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof, can be used to induce sedation or anesthesia. In certain embodiments, the compound of Formula (I), or pharmaceutically acceptable salt thereof, is useful in the treatment or prevention of adjustment disorders, anxiety disorders, cognitive disorders, dissociative disorders, eating disorders, mood disorders, schizophrenia or other psychotic disorders, sleep disorders, substance-related disorders, personality disorders, autism spectrum disorders, neurodevelopmental disorders, sterol synthesis disorders, pain, seizure disorders, stroke, traumatic brain injury, movement disorders and vision impairment, hearing loss, and tinnitus. In some embodiments, the disorder is Huntington's disease. In some embodiments, the disorder is Parkinson's disease. In some embodiments, the disorder is an inflammatory disease (e.g., lupus).

[0189] In another aspect, provided is a method of treating or preventing brain excitability in a subject susceptible to or afflicted with a condition associated with brain excitability, comprising administering to the subject an effective amount of a compound of the present invention. e.g., a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

[0190] In yet another aspect, the present invention provides a combination of a compound of the present invention. e.g., a compound of Formula (I), or pharmaceutically acceptable salt thereof, and another pharmacologically active agent. The compounds provided herein can be administered as the sole active agent or they can be administered in combination with other agents. Administration in combination can proceed by any technique apparent to those of skill in the art including, for example, separate, sequential, concurrent and alternating administration.

Diseases and Disorders

[0191] Described herein are methods of treating a sterol synthesis disorder. Exemplary disorders are described herein. The methods include administering to a subject. e.g., a subject suffering from a sterol synthesis disorder such as SLOS, a NMDA receptor modulating compound. Exemplary compounds are described herein.

Sterol Synthesis Disorders

[0192] In one aspect, described herein are methods for treating a sterol synthesis disorder. Cholesterol has an essential rule in growth and development. It is a membrance lipid and a precursor to many molecules that play important roles in cellular growth and differentiation, protein glycosylation, and signaling pathways. Biosynthesis of cholesterol involves a number of enzymes and intermediates. Disorders resulting from a deficiency in any of the enzymes involved in cholesterol biosynthesis lead to the accumulation of intermediates and imbalance in biomolecules, resulting in disorders including congenital skeletal malformations, dysmorphic facial features, psychomotor retardation, and failure to thrive. In an embodiment, a sterol synthesis disorder or symptom of a sterol synthesis disorder can be treated by administering to a subject suffering from a sterol synthesis disorder a compound described herein, such as a NMDA receptor modulating compound as described herein. Additional disorders are described below.

Smith-Lemli-Opitz Syndrome

[0193] In one aspect, described herein are methods for treating Smith-Lemli-Opitz Syndrome (or SLOS, or 7-dehydrocholesterol reductase deficiency). SLOS is an inborn error of cholesterol synthesis. In addition to microcephaly, moderate to severe intellectual disability, sensory hypersensitivity, stereotyped behaviors, dysmorphic facial features, and syndactyly of the second/third toes, a feature of the disease is reduced cerebrosterol (24(S)-hydroxycholesterol) levels. SLOS is an autosomal recessive genetic condition resulting from deficiency in the final enzyme of the cholesterol synthesis pathway, and causes low or low-normal plasma cholesterol levels and increased 7- and 8-dehydrocholesterol (DHC; 7DHC and 8DHC) levels. Common therapies currently used include dietary cholesterol supplementation, treatment with 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (HMG CoA reductase inhibitors, also known as statins), and treatment with agents that enhance cholesterol production and/or accretion; and to decrease the accumulation of 7DHC and 8DHC, the potentially toxic precursors of cholesterol.

Desmosterolosis

[0194] Desmosterolosis is a deficiency in desmosterol reductase and has a similar phenotype to SLOS. In one aspect, described herein are methods for treating desmosterolosis with compounds described herein.

Sitosterolemia

[0195] Sitosterolemia is a rare autosomal recessive disorder caused by mutations in two ATP-binding cassette (ABC) transporter genes (ABCG5 and ABCG8). Sitosterolemia enhances the absorption of plant sterols and cholesterol from the intestines. Patients typically present with tendon and tuberous xanthomas and premature coronary artery disease. In one aspect, described herein are methods for treating sitosterolemia with compounds described herein.

Cerebrotendinous Xanthomatosis (CTX)

[0196] In one aspect, described herein are methods for treating cerebrotendinous xanthomatosis (also referred to as cerebral cholesterosis, or Van Bogaert-Scherer-Epstein syndrome) with compounds described herein. CTX can be caused by a mutation in the CYP27A1 gene, which produces the sterol 27-hydroxylase enzyme. Sterol 27-hydroxylase metabolizes cholesterol into bile acids (e.g., chenodeoxycholic acid) that are important in the absorption of fats in the intestine. Enzyme dysfunction can lead to cholesterol accumulation in tissues. CTX is characterized by childhood diarrhea, cataracts, tendon xanthomas, reduced mental capability and abnormal movements in adults.

Mevalonate Kinase Deficiency Syndromes (MKD)

[0197] Mevalonate Kinase Deficiency (also referred to as mevalonic aciduria (a more severe form of MKD), or Hyper IgD Syndrome (HIDS, or hyperimmunoglobulinemia D) with period fever syndrome (a more benign form of MKD)) causes an accumulation of mevalonic acid in the urine as a result of insufficient activity of mevalonate kinase. MKD can result in developmental delay, hypotonia, anemia, hepatosplenomegaly, dysmorphic features, mental retardation, and overall failure to thrive. Mevalonic aciduria is characterized by delayed physical and mental development, failure to thrive, recurrent episodes of fever with vomiting and diarrhea, enlarged liver, spleen and lymph nodes, microcephaly (small head size), cataract, low muscle tone, short statute, distinct facial features, ataxia, and anemia. HIDS is characterized by recurrent episodes of fever associated with swollen lymph nodes, joint pain, gastrointestinal issues and skin rash. In one aspect, described herein are methods for treating MKD with the compounds described herein.

SC4MOL Gene Mutation (SMO Deficiency)

[0198] SC4MOL gene deficiency is a genetic disorder in the cholesterol biosynthesis pathway (e.g., mutations in the SC4MOL gene encoding a novel sterol oxidase). SC$MOL deficiency is characterized by the accumulation of dimethyl and monomethyl sterols that can be detected in blood, skin flakes or primary skin fibroblasts. In one aspect, described herein are methods for treating SMO deficiency with compounds described herein.

Niemann-Pick Disease

[0199] Niemann-Pick disease is a lysosomal storage disease resulting from a genetic mutation that affects metabolism. Niemann-Pick disease leads to abnormal accumulation of cholesterol and other fatty substances (lipids) due to an inability of the body to transport the substances. The accumulation damages the affected areas.

Autism

[0200] In one aspect, described herein are methods for treating autism spectrum disorder or autism. Autism spectrum disorder (ASD) and autism refer to a group of complex disorders of brain development. Autism is typically characterized by difficulties in social interaction, for example in verbal and nonverbal communication. Repetitive behaviors are also often seen in individuals having autism. Autism can be associated with intellectual disability, difficulties in motor coordination and attention and physical health issues. e.g., sleep and gastrointestinal disturbances. Individuals having autism can also excel in visual skills, music, math and art. Autism can refer to autistic disorder, childhood disintegrative disorder, pervasive developmental disorder—not otherwise specified (PDD-NOS), and Asperger syndrome. Autism also refers to monogenetic causes of autism such as synaptophathy's, e.g., Rett syndrome, Fragile X syndrome, Angelman syndrome.

Disorders Associated with Phenylketonuria

[0201] In one aspect, described herein are methods for treating disorders associated with phenylketonuria (e.g., cognitive disorders) with compounds described herein. Phenylketonuria can lead to hypochesterolemia and lowered vitamin D status. Total and low-density cholesterols and 25-hydroxy vitamin D have been found to be decreased in subjects suffering from phenylketonuria as compared with subjects not suffering from phenylketonuria (Clin. Chim. Acta 2013, 416: 54-59), 24S-hydroxycholesterol and 27S-hydroxycholesterol and 7α-hydroxycholesterol (e.g., representing peripheral and hepatic cholesterol elimination, respectively) have been shown to be significantly decreased in subjects suffering from phenylketonuria, while 7β-hydroxycholesterol (e.g., reflecting oxidative stress) was increased significantly in subjects suffering from phenylketonuria. Changes in the levels of 24S-OHC and 7β-hydroxycholesterol correlate with phenylalanine level, and 27S-hydroxycholesterol levels may correlate with the 25-hydroxy vitamin D level in subjects suffering from phenylketonuria.

Abbreviation

[0202] DCC: dicyclohexylcarbodiimide; DMAP: 4-dimethylaminopyridine; TEA: triethylamine; AlaOH: alanine; Boc: t-butoxycarbonyl. Py: pydidine; THF: tetrahydrofuran; TMS: trimethylsilyl; TBS (TBDMS): t-butyldimethylsilyl; Na.sub.2SO.sub.4: sodium sulfate; PE: petroleum ether, DCM: dichloromethane; EtOAc: ethylacetate, MeOH: methanol; Py: pyridine, Boc: t-butoxycarbonyl; MTBE: methyl tert-butyl ether.

EXAMPLES

[0203] In order that the invention described herein may be more fully understood, the following examples are set forth. Synthetic methods or intermediates may be found, for example in WO2014/160480*. 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.

[0204] The stereochemistry assigned herein (e.g., the assignment of “R” or “S” to the C24 position of the steroid) may be tentatively (e.g., randomly) assigned. For example, a C24 position may be drawn in the “R” configuration when the absolute configuration is “S.” A C24 position may also be drawn in the “S” configuration when the absolute configuration is “R.”

Example 1. Synthesis of Compound 1

[0205] ##STR00028##

[0206] Synthesis of Compound 1-2. To a solution of Compound 1-1 (0.3 g, 0.58 mmol; synthesized as described in Takahashi et al., Tetrahedron Letters, 2003, 44(2), 341-344) in DCM (10 mL) was added DMAP (7.08 mg, 0.058 mmol). TEA (95.7 mg, 0.87 mmol). DCC (179 mg, 0.87 mmol) and 2-((tert-butoxycarbonyl)amino)acetic acid (203 mg, 1.16 mmol). The mixture was stirred at 15° C. for 16 hours, at which point an additional aliquot of DMAP (7.08 mg, 0.058 mmol), TEA (95.7 mg, 0.87 mmol), DCC (179 mg, 0.87 mmol) and 2-((tert-butoxycarbonyl)amino)acetic acid (203 mg, 1.16 mmol) were added. The mixture was stirred at 25° C. for 16 hours, then the mixture was filtered and the filtrate was diluted with aqueous sat. NH.sub.4Cl (30 mL) and extracted with DCM (10 mL×2). The combined organic phases were dried over Na.sub.2SO.sub.4, filtered, and concentrated to give the crude product Compound 1-2, which was used in the next step directly without purification. .sup.1H NMR indicated an estimated yield of 45%.

[0207] Synthesis of Compound 1-3. Compound 1-2 (350 mg, 0.519 mmol) was dissolved in TBAF (5.18 mL, 5.18 mmol, 1M in THF) and the mixture was stirred at 15° C. for 16 hours. The mixture was quenched with saturated NH.sub.4Cl (10 mL) and extracted with EtOAc (5 mL×2). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated, and purified by silica gel (PE: EtOAc=10:1) to give the crude product Compound 1-3 (250 mg. Compound 1-2/Compound 1−3=0.55/0.45) product as a colorless oil.

[0208] Synthesis of Compound 1. Compound 1-3 was dissolved in HC/dioxane (4 N, 5 mL), and the mixture was stirred at 15° C. for 30 minutes. Then MTBE (5 mL) was then added to the mixture to form a precipitate, which was filtered and purified by prep-HPLC (column: Phenomenex Synergi C18 150*30 mm*4 um, gradient: 36-66% B (A=0.05% HCl-ACN, B=acetonitrile) flow rate: 30 mL/min) to afford the HCl salt of Compound 1 (6 mg, 2.92%) as an off white solid. .sup.1H NMR (400 MHz, MeOD) δ 5.36-5.35 (m, 1H), 3.91-3.81 (m, 2H), 3.43-3.38 (m, 1H), 2.68-2.25 (m, 2H), 2.10-0.94 (m, 37H), 0.74 (s, 3H). LCMS MS ESI calcd. for C.sub.29H.sub.49NO.sub.3Na [M+Na].sup.+482, found 482.

Example 2. Synthesis of Compound 2

[0209] ##STR00029##

[0210] Synthesis of Compound 2-2. To a solution of Compound 1-1 (0.3 g, 0.58 mmol) in DCM (10 mL) was added DMAP (21.2 mg, 0.174 mmol), TEA (191 mg, 1.74 mmol), DCC (452 mg, 1.74 mmol) and (S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid (378 mg, 1.74 mmol). The mixture was stirred at 25° C. for 16 hours. The mixture was filtered, the filtrate was concentrated and purified by combi-flash (PE: EA=100%-95%) to give Compound 2-2 (380 mg, 92%) as a colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.31-5.19 (m, 1H), 5.03-4.99 (m, 1H), 4.74-4.72 (m, 1H), 4.23-4.20 (m, 1H), 3.50-3.44 (m, 1H), 2.29-2.17 (m, 3H), 1.85-0.85 (m, 60H), 0.66 (s, 3H), 0.05 (s, 6H).

[0211] Synthesis of Compound 2-3. Compound 2-2 (380 mg, 0.53 mmol) in TBAF (15.9 mL, 15.9 mmol, 1M in THF) was stirred at 15° C. for 16 hours. The mixture was quenched with aqueous sat. NH.sub.4Cl (20 mL) and extracted with EtOAc (10 mL×2). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated, and purified by combi-flash (PE: EA=100%-90%) to give Compound 2-3 as a colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.35-5.29 (m, 1H), 5.01-4.99 (m, 1H), 4.75-4.73 (m, 1H), 4.23-4.20 (m, 1H), 3.52-3.50 (m, 1H), 2.29-2.17 (m, 3H), 1.85-0.85 (m, 52H), 0.66 (s, 3H).

[0212] Synthesis of Compound 2. Compound 2-3 (50 mg, 0.083 mmol) was dissolved in HCl/dioxane (1 mL) and stirred at 15° C. for 30 minutes. Then MTBE (1 mL) was then added to the mixture and a precipitate was formed, which was filtered to afford the HCl salt of Compound 2 (5 mg, 12.0%) as an off-white solid. .sup.1H NMR (400 MHz, MeOD) δ 5.36-5.30 (m, 1H), 3.97-3.80 (m, 1H), 3.50-3.40 (m, 1H), 2.50-2.40 (m, 3H), 2.35-0.85 (m, 46H), 0.74 (s, 3H). LCMS ESI calcd. for C.sub.32H.sub.56NO.sub.3 [M+H].sup.+ 502, found 502.

Example 3. Synthesis of Compound 3

[0213] ##STR00030##

[0214] Synthesis of 3-2. Iodine (1.17 g, 4.63 mmol) was added to a solution of trimethyl phosphite (526 mg, 4.24 mmol) in CH.sub.2Cl.sub.2 (20 mL) at 0° C. After stirring for 5 minutes, the clear colorless solution was allowed to warm to 15° C. The phosphorylating agent was added dropwise to a solution of Compound 1-1 (2.0 g, 3.86 mmol) and pyridine (1.21 g, 15.4 mmol) in CH.sub.2Cl.sub.2 (30 mL) at 0° C. The resulting mixture was stirred at 0° C. for 1 h, then cooled to 15° C. and stirred for another 1 h. The reaction mixture was treated water (50 mL), extracted with CH.sub.2Cl.sub.2 (50 mL×2). The combined organic phase was washed with brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. The residue was purified by column chromatography on silica gel (PE/EtOAc=10/1 to 3/1) to afford Compound 3-2 (1.8 g, 75%) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.35-5.30 (m, 1H), 4.20-4.17 (m, 1H), 3.76 (s, 3H), 3.73 (s, 3H), 3.48-3.46 (m, 1H), 2.26-2.15 (m, 2H), 1.93-1.61 (m, 7H), 1.55-0.88 (m, 38H), 0.66 (s, 3H), 0.05 (s, 6H).

[0215] Synthesis of Compound 3. To a solution of Compound 3-2 (300 mg, 480 μmol) in CH.sub.2Cl.sub.2 (5 mL) was added bromotrimethylsilane (220 mg, 1.44 mmol) at 15° C. and the reactions was stirred for 12 h. The reaction mixture was adjusted to pH=8 with aq. NaOH (1.44 mL, 1.44 mmol, 1 M in H.sub.2O) and a precipitate was formed, which was then filtered, washed with CH.sub.2Cl.sub.2 (2 mL) and water (2 mL), and dried under vacuum to afford Compound 3 (30 mg, 12%) as an off-white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 5.24-5.23 (m, 1H), 3.87-3.85 (m, 1H), 3.24-3.21 (m, 1H), 2.13-2.06 (m, 2H), 1.91-1.60 (m, 6H), 1.50-0.80 (m, 33H), 0.62 (s, 3H). LCMS MS ESI calcd. for C.sub.27H.sub.44 [M+H—H.sub.2O—Na.sub.2O—Na.sub.2O.sub.4P].sup.+ 367, found 367. HRMS MS ESI calcd. for C.sub.27H.sub.46O.sub.5P [M−H].sup.− 481.3088, found 481.3105.

Example 4. Synthesis of Compound 4

[0216] ##STR00031##

[0217] Synthesis of Compound 4-2. To a solution of Compound 4-1* (2 g, 5.01 mmol) and Pd/C (200 mg, 10%) in THF (30 mL) was hydrogenated under 15 psi of hydrogen at 25° C. for 3 h. The mixture was filtered through a pad of celite and the filtrate was concentrated in vacuum to afford Compound 4-2 (1.8 g. crude) as an off-white solid.

[0218] Synthesis of Compound 4-3. To a solution of Compound 4-2 (1.8 g, 4.47 mmol) in THF (25 mL) was added a solution LiAlH.sub.4 (339 mg, 8.94 mmol) in THF (5 mL) drop wise below 15° C. The solution was stirred at 15° C. for 2 h. The reaction was quenched by the addition of saturated aqueous NH.sub.4C (20 mL) at 0° C. The resulting mixture was extracted with EtOAc (2×50 mL). The combined organic layer was washed with brine (2×30 mL) and concentrated in vacuum to afford Compound 4-3 (1.6 g, crude) as a light yellow solid.

[0219] Synthesis of Compound 4-4. A mixture of Compound 4-3 (1.6 g, 4.27 mmol) in DCM (10 mL) and THF (10 mL) was added PCC (2.27 g, 10.6 mmol) at 25° C. The reaction was stirred at 25° C. for 3 hrs. The solution was filtered and the filter cake was washed with DCM (25 mL). The combined filtrate was concentrated in vacuum. The residue was purified by silica gel column, eluting with PE/EtOAc=8/1 to give Compound 4-4 (0.9 g, 54%) as an off-white solid.

[0220] Synthesis of Compound 4-5. To a solution of Compound 4-4 (0.9 g, 2.41 mmol) in THF (30 mL) was added drop wise isopropyl magnesium chloride (3.61 mL, 7.23 mmol, 2M in THF) at −78° C. The mixture was stirred at −78° C. for 2 hrs. Then, the mixture was allowed to warm up to 25° C. and stirred for 3 hrs. The reaction was poured into water (100 mL) and extracted with EtOAc (2×30 mL). The combined organic layer was washed with brine (50 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuum. The residue was purified by silica gel column, eluting with PE/EtOAc=5/1 to afford Compound 4-5 (0.6 g, 57%) as an off-white solid.

[0221] Synthesis of Compound 4. To a solution of Compound 4-5 (200 mg, 479 μmol) in pyridine (3 mL) was added SO.sub.3—Py (76.1 mg, 4.79 mmol), and the mixture was stirred at 40° C. for 16 hours. Pyridine was removed under reduced pressure and the residue was diluted with NaOH (3% in water, 20 mL), then extracted with BuOH (10 mL×3). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated, and purified by combi-flash (DCMin MeOH=100%-75%) to give Compound 4 (6 mg, 2%) as an off-white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 5.25-5.19 (m, 1H), 4.35 (brs, 1H), 3.85-3.75 (m, 1H), 1.95-0.79 (m, 41H), 0.65 (m, 3H). LCMS MS ESI calcd. for C.sub.28H.sub.47O.sub.4S [M+H—H.sub.2O].sup.+ 479, found 479.

Example 5. Synthesis of Compound 5

[0222] ##STR00032##

[0223] Synthesis of Compound 5-2A and Compound 5-2B. Molecular iodine (1.10 g, 4.34 mmol) was added to a solution of trimethyl phosphite (584 mg, 4.71 mmol) in CH.sub.2Cl.sub.2 (3 mL) at 0° C. After stirring for 5 minutes, the clear, colorless solution was warmed to 20° C. and added dropwise to a solution of Compound 5-1 (500 mg, 1.24 mmol) and pyridine (783 mg, 9.92 mmol) in CH.sub.2Cl.sub.2 (2 mL) at 0° C. The resulting mixture was stirred at 0° C. for 1 h, then warmed to 20° C. and stirred for another 1 h. The reaction mixture was treated with water (20 mL) and extracted with CH.sub.2Cl.sub.2 (20 mL×2). The combined organic phase was washed with brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated under vacuum, and purified on silica gel (PE/EtOAc=8/1 to 3/1) to afford Compound 5-2A (200 mg, 32%) as an off-white solid and Compound 5-2B (50 mg, 7%) as a light yellow oil. Compound 5-2A: .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.39-5.35 (m, 1H), 4.30-4.18 (m, 2H), 3.77 (s, 6H), 3.74 (s 6H), 2.44-2.40 (m, 2H), 1.98-1.57 (m, 8H), 1.48-0.92 (m, 28H), 0.67 (s, 3H). Compound 5-2B: .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.39-5.36 (m, 1H), 4.25-4.18 (m, 1H), 3.77 (s, 3H), 3.74 (s, 3H), 3.35-3.25 (m, 1H), 2.44-2.40 (m, 2H), 1.99-1.57 (m, 8H), 1.57-0.88 (m, 29H), 0.68 (s, 3H).

[0224] Synthesis of Compound 5. To a solution of Compound 5-2A (50 mg, 80.8 μmolμmol) in CH.sub.2Cl.sub.2 (3 mL) was added TMSBr (74.1 mg, 484.8 μmolμmol) at 20° C. and the mixture stirred for 12 his, at which point TLC analysis indicated the starting material was consumed completely. The reaction mixture was adjusted to pH=8 with aq. NaOH (484 uL, 484 μmol, 1 M in H.sub.2O) and a precipitate was formed. The white solid was filtered and washed with CH.sub.2Cl.sub.2 (2 mL) and water (2 mL), and dried under vacuum to afford Compound 5 (9.8 mg, 19%) as an off-white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 5.30-5.25 (m, 1H), 3.95-3.80 (m, 2H), 2.37-2.26 (m, 1H), 2.26-2.15 (m, 1H), 1.96-1.77 (m, 6H), 1.55-0.79 (m, 30H), 0.62 (s, 3H). LCMS MS ESI calcd. for C.sub.27H.sub.43 [M+H-2H.sub.3PO.sub.4].sup.+ 367, found 367. HRMS MS ESI calcd. for C.sub.27H.sub.47O.sub.8P.sub.2 [M−H].sup.− 561.2752, found 561.2760.

Example 6. Synthesis of Compound 6

[0225] ##STR00033##

[0226] Synthesis of Compound 3-2. Iodine (1.17 g, 4.63 mmol) was added to a solution of trimethyl phosphite (526 mg, 4.24 mmol) in CH.sub.2Cl.sub.2 (20 mL) at 0° C. After stirring for 5 minutes, the clear, colorless solution was allowed to warm to 15° C. after which it was added dropwise to a solution of Compound 1-1 (1.21 g, 15.4 mmol) in CH.sub.2Cl.sub.2 (30 mL) at 0° C. The resulting mixture was stirred at 0° C. for 1 h and then warmed to 15° C. and stirred for another 1 h. The reaction mixture was treated water (50 mL), extracted with CH.sub.2Cl.sub.2 (50 mL×2), and the combined organic phase was washed with brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. The residue was purified on silica gel (PE/EtOAc=10/1 to 3/1) to afford Compound 3-2 (1.8 g, 75%) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.35-5.30 (m, 1H), 4.20-4.18 (m, 1H), 3.76 (s, 3H), 3.73 (s, 3H), 3.48-3.46 (m, 1H), 2.26-2.17 (m, 2H), 1.95-1.61 (m, 7H), 1.48-0.88 (m, 41H), 0.66 (s, 3H), 0.05 (s, 6H).

[0227] Synthesis of Compound 6-3A and Compound 6-3B. Compound 3-2 (1.5 g, 2.40 mmol) was dissolved in MeOH (20 mL) and separated by supercritical fluid chromatography (SFC) to afford Compound 6-3A (490 mg, 33%) and Compound 6-3B (400 mg, 27%) as off-white solids. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.30-5.25 (m, 1H), 4.20-4.18 (m, 1H), 3.76 (s, 3H), 3.73 (s, 3H), 3.48-3.44 (m, 1H), 2.30-2.10 (m, 2H), 1.98-1.57 (m, 7H), 1.53-0.88 (m, 38H), 0.67 (s, 3H), 0.05 (s, 6H). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.30-5.25 (m, 1H), 4.20-4.17 (m, 1H), 3.76 (s, 3H), 3.73 (s, 3H), 3.50-3.44 (m, 1H), 2.26-2.10 (m, 2H), 1.98-1.62 (m, 11H), 1.48-0.88 (m, 34H), 0.67 (s, 3H), 0.05 (s, 6H). For 6-3B, 91% de was obtained that was not subjected to deprotection.

[0228] Synthesis of 6. To a solution of Compound 6-3A (100 mg, 160 mol) in CH.sub.2Cl.sub.2 (3 mL) was added TMSBr (97.9 mg, 640 μmol) and the reaction was stirred f at 20° C., or 12 h. The reaction mixture was adjusted to pH=8 with aq. NaOH (640 uL, 640 μmol, 1 M in H.sub.2O) and the solid was precipitated. The white solid was filtered and washed with CH.sub.2Cl.sub.2 (2 mL), water (2 mL), dried by vacuum to provide Compound 6 (45.7 mg, 54%) as an off-white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 5.30-5.20 (m, 1H), 3.89-3.80 (m, 1H), 3.26-3.20 (m, 1H), 2.15-1.60 (m, 8H), 1.55-0.80 (m, 30H), 0.63 (s, 3H). LCMS Rt=1.330 min in 2.0 min chromatography, 10-80 AB, MS ESI calcd. for C.sub.27H.sub.43 [M+H—H.sub.3PO.sub.4—H.sub.2O].sup.+ 367, found 367. HRMS MS ESI calcd. for C.sub.27H.sub.46O.sub.5P [M−H].sup.− 481.3088, found 481.3085.

Example 7. Synthesis of Compound 7

[0229] ##STR00034##

[0230] To a solution of Compound 4-1 (0.1 g, 0.239 mmol) in pyridine (2 mL) was added dihydrofuran-2,5-dione (71.7 mg, 0.717 mmol) and DMAP (14.5 mg, 0.119 mmol). The mixture was stirred at 15° C. for 16 hours. To the reaction was added aqueous sat. NH.sub.4Cl (10 mL) and extracted with EtOAc (2×5 mL). The combined organic phase was dried over Na.sub.2SO.sub.4 filtered, concentrated and purified by combi-flash (PE: EA=100%-70%. DCM: MeOH=100%-95%) to give Compound 7 (40 mg) as colorless oil, which was washed with PE (5 mL) and filtered to give Compound 7 (10 mg, yield 8%) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.30-5.29 (m, 1H), 4.73-7.70 (m, 1H), 2.67-2.63 (m, 4H), 2.45-2.35 (m, 1H), 1.99-1.95 (m, 3H), 1.85-0.80 (m, 39H), 0.66 (s, 3H). LCMS t.sub.R=1.591 min in 2 min chromatography, 10-80AB_ELSD, MS ESI calcd. for C.sub.32H.sub.52O.sub.5Na [M+Na].sup.+ 539, found 539.

Example 10. Synthesis of Compound 10

[0231] ##STR00035##

[0232] To a solution of Compound 10-1 (50 mg, 0.124 mmol; synthesized as described in Upasani et al., WO2013/36835, [00375]) in pyridine (2 mL) was added dihydrofuran-2,5-dione (37.2 mg, 0.372 mmol) and DMAP (7.57 mg, 0.062 mmol). The mixture was stirred at 15° C. for 16 hours and then another batch of dihydrofuran-2,5-dione (37.2 mg, 0.372 mmol) and DMAP (7.57 mg, 0.062 mmol) was added. The mixture was stirred at 15° C. for 16 hours. The reaction solution was quenched with aqueous sat. NH.sub.4Cl (5 mL) and extracted with EtOAc (3 mL×2). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated. The residue was purified by combi-flash (0˜30% of EtOAc in PE) to give 50 mg of impure Compound 10. The mixture was further purified by prep-HPLC (column: Gemini 150*25 5 u, gradient: 65 B to give Compound 10 (5 mg, 8% yield) as an off-white solid. .sup.1H NMR (400 MHz, CDCl3) δ 5.35-5.25 (m, 1H), 2.60-2.55 (m, 5H), 2.45-2.35 (m, 2H), 2.10-0.80 (m, 39H), 0.67 (s, 3H). LCMS t.sub.R=1.523 min in 2 min chromatography, 10-80AB_ELSD, MS ESI calcd. for C.sub.31H.sub.50O.sub.5Na [M+Na].sup.+ 525, found 525.

Example 11. Synthesis of Compound 11

[0233] ##STR00036##

[0234] Synthesis of Compound 11-2. To a solution of Compound 5-1 (300 mg, 745 μmol) and (S)-2-((tert-butoxycarbonyl)amino)propanoic acid (423 mg, 2.41 mmol) in DCM (5 mL) was added DCC (767 mg, 3.72 mmol), DMAP (45 mg, 368 μmol), TEA (452 mg, 4.47 mmol). The mixture was stirred at 30° C. for 3 hours. The mixture was washed by brine (10 mL) and extracted by EtOAc (20 mL×2). The combined organic layer was dried by Na.sub.2SO.sub.4, filtered and evaporated in vacuum. The residue was purified by silica gel chromatography (PE/EtOAc=5/1) to afford Compound 11-2 (300 mg, 54% yield) as a yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.37 (m, 1H), 5.17-5.04 (m, 2H), 4.76-4.61 (m, 2H), 4.30-4.17 (m, 2H), 2.36-2.30 (m, 2H), 2.00-1.95 (m, 2H), 1.88-1.77 (m, 4H), 1.61-0.88 (m 54H), 0.67 (s, 3H).

[0235] Synthesis of Compound 11. Compound 11-2 (150 mg, 209 μmol) was added to HCl/dioxane (3 mL, 4M). The mixture was stirred at 25-27° C. for 1 hour. The mixture was filtered and washed with MTBE (5 mL×2), dried in vacuum to give an off-white solid. The solid was dissolved in water/MeCN (5 mL/1 mL) and lyophilized 3 times to remove residual solvent to afford Compound 11 (71 mg, yield 43% yield) as an off-white solid, 1H NMR (400 MHz, MeOD) δ5.45-5.44 (m, 1H), 4.74-4.66 (m, 1H), 4.17-4.05 (m, 2H), 2.44-2.37 (m, 2H), 2.09-1.87 (m, 6H), 1.72-0.95 (m, 37H), 0.75 (s, 3H). LCMS Rt=0.831 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.33H.sub.57N.sub.2O.sub.4 [M+H].sup.+ 545, found 545.5.

Example 12. Synthesis of Compound 12

[0236] ##STR00037##

[0237] Synthesis of Compound 12-2. To a solution of Compound 1-1 (500 mg, 967 μmol) in DCM (5 mL) was added (S)-2-((tert-butoxycarbonyl)amino)propanoic acid (586 mg, 3.10 mmol). DCC (764 mg, 3.71 mmol). TEA (376 mg, 3.72 mmol), DMAP (151 mg, 1.24 mmol). The mixture was stirred at 30° C. for 3 hours. The mixture was washed by water (10 mL) and extracted with EtOAc (20 mL×2). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated to give a residue, which was diluted with water (15 mL). The suspension was heated at 60° C. for 30 minutes. The mixture was filtered to give Compound 12-2 (579 mg, 1.00 mmol) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.31 (d, 1H), 5.19 (s, 1H), 5.08 (d, 1H), 4.79-4.62 (m, 1H), 4.38-4.06 (m, 1H), 3.56-3.39 (m, 1H), 2.35-2.10 (m, 2H), 2.08-1.90 (m, 2H), 1.90-1.66 (m, 5H), 1.65-1.32 (m, 21H), 1.30-0.79 (m, 28H), 0.66 (s, 3H), 0.17-0.01 (m, 6H).

[0238] Synthesis of Compound 12-3. To a solution of Compound 12-2 (300 mg, 435 μmol) was added TBAF/THF (5 mL, 1 M). The mixture was stirred at 23-25° C. for 30 minutes. The mixture was diluted with water (20 mL) and extracted by EtOAc (20 mL×2). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated to give a residue, which was purified by SFC ((Column: AD (250 mm*30 mm, 5 um). Condition: Base-ETOH, Begin: B 25% FlowRate (ml/min): 70) to give Compound 12-3 (392 mg) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.53-5.20 (m, 1H), 5.09 (d, 1H), 4.86-4.57 (m, 2H), 4.30 (t, 1H), 3.84-3.35 (m, 1H), 2.52-2.17 (m, 2H), 2.04-1.92 (m, 2H), 1.89-1.75 (m, 4H), 1.69-0.82 (m, 42H), 0.67 (s, 3H).

[0239] Synthesis of Compound 12. Compound 12-1 (300 mg, 522 μmol) was added to HCl/dioxane (3 ml, 4M). The mixture was stirred at 25-27° C. for 1 hour. The mixture was filtered and washed with MTBE (5 ml×2), dried in vacuum to give an off-white solid. The residue was dissolved in water/MeCN (5 mL/1 mL) and lyophilized 3 times to removal residual solvents to give Compound 12-2 (90 mg, 34% yield) as an off white solid. .sup.1H NMR (400 MHz, MeOD) δ 5.36 (d, 1H), 4.95-4.4.70 (m, 3H), 3.80-3.70 (m, 1H), 3.46-3.36 (m, 1H), 2.28-2.16 (m, 2H), 2.10-1.75 (m, 6H), 1.76-1.61 (m, 2H), 1.62-1.39 (m, 11H), 1.38-1.25 (m, 1H), 1.25-1.07 (m, 4H), 1.04 (s, 3H), 1.01-0.91 (m, 10H), 0.74 (s, 3H). LCMS Rt=0.991 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.30H.sub.51NO.sub.3Na [M+Na].sup.+ 496.39, found 496.3.

Example 13. Synthesis of Compound 13

[0240] ##STR00038##

[0241] Step 1. To a solution of 13-1 (7.0 g, 16.2 mmol) in THF (70 mL) was added dropwise ethylmagnesium bromide (26.9 mL, 80.9 mmol, 3M in Et.sub.2O) at 0° C. under nitrogen atmosphere. The reaction mixture was stirred at 25° C. for 12 h. TLC showed the starting material was consumed completely. The mixture was quenched with saturated aqueous NH.sub.4Cl (100 mL) and extracted with EtOAc (200 mL×2). The combined organic phase was washed with brine (200 mL), dried over anhydrous Na.sub.2SO.sub.4, concentrated in vacuum. The residue was purified by column chromatography on silica gel (PE/EtOAc=20/1) to afford 13-2 (500 mg, 7.7%) as white solid. .sup.1H NMR (400 MHz, CDCl3) δ 5.30-5.28 (m, 1H), 2.42-2.39 (m, 5H), 1.98-1.63 (m, 8H), 1.53-1.25 (m, 16H), 1.23-0.84 (m, 28H), 0.66 (s, 3H).

[0242] Step 2. To a solution of 13-2 (500 mg, 1.24 mmol) in MeOH (10 mL) was added NaBH. (93.8 mg, 2.48 mmol) in portions. The reaction mixture was stirred at 25° C. for 2 h. After TLC showed the starting material was consumed and a new spot was produced. The reaction mixture was quenched with saturated aqueous NH.sub.4Cl (10 mL), extracted with EtOAc (30 mL×2). The combined organic phase was washed with brine (30 mL), concentrated in vacuum. The residue was purified by column chromatography on silica gel (PE/EtOAc=20/1) to afford 13-3 (500 mg, crude), which was purified by prep-HPLC to afford the pure 13-3 (60 mg, 12%) as white solid. .sup.1H NMR (400 MHz, CDCl3) δ 5.29 (d, J=4.4 Hz, 1H), 3.48-3.47 (m, 1H), 2.42 (d, J=12.8 Hz, 1H), 2.02-1.57 (m, 12H), 1.57-0.92 (m, 26H), 0.67 (s, 3H).

[0243] Step 3. To a solution of Compound 13-3 (200 mg, 496 μmol) in pyridine (2 mL) was added DMAP (30.3 mg, 248 μmol) and dihydrofuran-2,5-dione (199 mg, 1.98 mmol). The mixture was stirred at 25-27° C. for 16 hr. The reaction mixture was washed by aqueous sat.NH.sub.4Cl (3 ml) and extracted with ethyl acetate (2 mL×2). The combined organic layer was concentrated in vacuum. The residue was purified by silica gel chromatography (PE/EtOAc=1/1) to afford Compound 13 (42 ng, 17% yield for mixture of diastereomers) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δδ 5.35-5.22 (m, 1H), 4.87-4.70 (m, 1H), 2.79-2.54 (m, 4H), 2.42 (d, 1H), 2.09-1.91 (m, 3H), 1.89-1.64 (m, 3H), 1.64-1.30 (m, 13H), 1.32-0.76 (m, 21H), 0.67 (s, 3H). LCMS Rt=2.265 min in 3.0 min chromatography, 10-80 AB, MS ESI calcd. for C.sub.31H.sub.50O.sub.5Na [M+Na].sup.+ 525.37, found 525.3.

Example 14. Synthesis of Compound 14

[0244] ##STR00039##

[0245] Synthesis of Compound 14-2. To a solution of Compound 1-1 (0.3 g, 0.58 mmol) in pyridine (2 mL) was added Py-SO.sub.3 (276 mg, 1.74 mmol). The mixture was stirred at 40° C. for 16 hours. The mixture was concentrated and the residue was diluted with NaOH (20 mL, 3% in water). The mixture was extracted with PE (10 mL) and the organic layer was separated. The aqueous layer was extracted with BuOH (2×15 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated to give Compound 14-2 (400 mg, crude) as yellow solid. .sup.1H NMR (400 MHz, MeOD) δ 5.34-5.33 (m, 1H), 4.14-4.11 (m, 1H) 3.58-3.54 (m, 1H), 2.30-0.85 (m, 48H), 0.74 (s, 3H), 0.08 (s, 6H).

[0246] Synthesis of Compound 14. To a solution of Compound 14-2 (200 mg, 0.335 mmol) in DCM (5 ml) was added HF-Py (2 mL). The mixture was stirred at 15° C. for 16 hours. The reaction mixture was diluted with NaOH solution (10 mL, 3% in water) and extracted with DCM (2×5 mL). The combined organic layer was washed with NaOH solution (10 mL, 3% in water), separated, dried over Na.sub.2SO.sub.4, filtered and concentrated to give a residue, which was purified by combi-fish (PE: EA=100%-60%, DCM: MeOH=100%-90%) to give 50 mg of Compound 14 as colorless oil. The oil was crystallized from H.sub.2O to give Compound 14 (10 mg, 6%) as an off-white solid. .sup.1H NMR (400 MHz, MeOD) δ 5.36-5.35 (m, 1H), 4.63 (s, 1H), 4.17-4.13 (m, 1H), 3.43-3.38 (m, 1H), 2.24-2.20 (m, 2H), 2.15-0.85 (m, 37H), 0.74 (s, 3H). LCMS t.sub.R=1.271 min in 2 min chromatography, 10-80AB_ELSD, MS ESI calcd. for C.sub.27H.sub.45O.sub.4S [M+H—H.sub.2O].sup.+ 465, found 465.

Example 16. Synthesis of Compounds 16 and 17

[0247] ##STR00040##

[0248] Synthesis of Compound 5-3. To a solution of Compound 5-1 (1.5 g, 3.72 mmol), Boc-Ala-OH (703 mg, 3.72 mmol), DMAP (45.4 mg, 0.37 mmol), TEA (376 mg, 3.72 mmol) in DCM (20 mL) was added DCC (767 mg, 3.72 mmol) at 15° C. The mixture was stirred at 15° C. for 20 hrs. The reaction mixture was filtered and the filtrate was concentrated in vacuum to give a crude product, which was purified by flash column (EtOAc in PE, 0-30% in 60 minutes) to give Compound 5-3 (600 mg, 28%).

[0249] Synthesis of Compounds 16 and 17. To a solution of Compound 5-3 (600 mg) in dioxane (4 mL) was added HCl (2 mL, 4 M in dioxane) at 15° C. The mixture was stirred at 15° C. for 16 hrs. MTBE (15 mL) was added and an off-white solid was produced. The mixture was filtered. The filtered cake was washed with MTBE, concentrated in vacuum to give an off-white solid (600 mg), which was purified by prep. HPLC (Column: Phenomenex Gemini 150*25 mm*10 um; condition: water (0.05% HCl)-ACN, 50-60% B in 10 mins, 100% B Hold Time (min): 4; FlowRate (ml/min): 25) to give Compound 16 (9 mg, 11%) and Compound 17 (33 mg, 40%). Compound 16: .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.40-8.30 (br, 3H), 5.45-5.30 (m, 1H), 4.70-4.50 (m, 1H), 4.23-4.10 (m, 1H), 4.10-3.98 (m, 1H), 3.10-3.00 (m, 1H), 2.40-2.20 (m, 2H), 2.03-1.71 (m, 5H), 1.67-10.88 (m, 28H), 0.86-0.78 (m, 6H), 0.66 (s, 3H). LCMS Rt=1.014 min in 2.0 min chromatography, 30-90 AB_E, MS ESI calcd. for C.sub.27H.sub.45O [M+H-AlaOH].sup.+ 385, found 385.

[0250] Compound 17: .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.40-8.30 (br, 3H), 5.45-5.30 (m, 1H), 4.70-4.50 (m, 1H), 4.23-4.10 (m, 1H), 4.10-3.98 (m, 1H), 3.10-3.00 (m 1H), 2.40-2.20 (m, 2H), 2.03-1.71 (m, 5H), 1.67-0.88 (m, 28H), 0.86-0.78 (m, 6H), 0.66 (s, 3H). LCMS Rt=1.011 min in 2.0 min chromatography, 30-90 AB_E. MS ESI calcd. C.sub.27H.sub.45O [M+H-AlaOH].sup.+ 385, found 385.

Example 17. Synthesis of Compound 18

[0251] ##STR00041##

[0252] Synthesis of Compound 5-4. To a solution of Compound 5-1 (100 mg, 248 μmol) in DCM (2 mL) at 1318° C. was added N,N-dimethylpyridin-4-amine (3 mg, 24.5 μmol), triethylamine (25 mg, 247 μmol), (S)-2-((tert-butoxycarbonyl)amino)propanoic acid (47 mg, 248 μmol) and N,N′-methanediylidenedicyclohexanamine (51 mg, 247 μmol). The reaction was stirred over 16 hrs at 20°. The reaction was filtered and filtrate was concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=20/1) to give the desired product (45 mg, 24%) as a solid. Synthesis of Compound 18. To a solution of Compound 5-4 (45 mg, 78.4 μmol) in dioxane (1 mL) was added HC/dioxane (4 M, 1 mL) at 15-28° C. The reaction mixture was stirred for 3 hrs. and then 5 mL of sat. NaHCO.sub.3 was added so the reaction was at pH=9. The mixture was extracted with EtOAc (3×10 mL). The combined organic layer was washed with brine (10 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (DCM/MeOH=15/1) to give Compound 18 (3.2 mg, 8%) as an off-white solid. .sup.1H NMR (CDCl.sub.3, 400 MHz): δ 5.40-5.39 (m, 1H), 4.67-4.64 (m, 1H), 3.56-3.54 (m, 1H), 3.34-3.33 (m, 1H), 2.35-2.33 (m, 2H), 2.05-1.90 (m, 2H), 1.88 (m, 3H), 1.67-0.91 (m, 37H), 0.70 (s, 3H). LCMS Rt=1.866 min in 3.0 min chromatography, 10-80 AB, MS ESI calcd. for C.sub.27H.sub.45O [M+H-AlaOH].sup.+ 385, found 385.

Example 18. Synthesis of Compound 19

[0253] ##STR00042##

Synthesis of Compound 18-2. To a solution of Compound 18-1 (100 mg, 0.225 mmol; synthesized as described in Martinez et al., WO2014/160480) in DCM (1 mL) was added DMAP (8.2 mg, 0.0675 mmol). Boc-Vla-OH (146 mg, 0.675 mmol). TEA (68.3 mg, 0.675 mmol) and DCC (139 mg, 0.675 mmol). The mixture was stirred at 25° C. for 16 hours. The mixture was concentrated under vacuum, purified by column chromatography on silica gel (PE/EtOAc=15:1) to give Compound 18-2 (80 mg, 55%) as an off-white solid.

[0254] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.35-5.20 (m 2H), 5.00-4.90 (m, 1H), 4.38-4.25 (m, 1H), 2.48-2.37 (m, 1H), 2.25-2.15 (m, 1H), 2.02-1.90 (m, 3H), 1.85-0.85 (m, 46H), 0.67 (s, 3H). Synthesis of Compound 19. To a solution of Compound 18-2 (70 mg, 0.124 mmol) in dioxane (0.5 mL) was added HCl/dioxane (1 mL, 4 M). The mixture was stirred at 25° C. for 2 hours and an off-white solid was formed. To the reaction mixture was added MTEB (10 mL) and the reaction was filtered. The solid was washed with MTEB (10 mL) and then dissolved in MeOH (10 mL). The MeOH solution was concentrated in vacuum to give Compound 19 (55.6 mg, 77%) as an off-white solid. .sup.1H NMR (400 MHz, methanol-d4) δ 5.60-5.50 (m, 1H), 5.34-5.25 (m, 1H), 4.16 (d, J=4.0 Hz, 1H), 2.48-2.30 (m, 2H), 2.05-1.40 (m, 16H), 1.35-0.90 (m, 23H), 0.74 (s, 3H). LCMS Rt=0.970 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.31H.sub.51F.sub.3NO.sub.3 [M+H].sup.+ 542, found 542.

Example 19. Synthesis of Compound 20

[0255] ##STR00043##

[0256] To a solution of Compound 18-1 (50 mg, 0.122 mmol) in pyridine (1 mL) was added DMAP (6.8 mg, 0.056 mmol) and dihydrofuran-2,5-dione (33.6 mg, 336 mmol). The mixture was stirred at 25° C. for 16 hours. The mixture was concentrated under vacuum, purified by column chromatography on silica gel (PE/EtOAc=1:1) to give Compound 20 (14.1 mg, yield 23%) as an off-white solid. .sup.1H NMR (400 MHz, CDCs) S 5.35-5.25 (m, 2H), 2.78-2.65 (m, 4H), 2.46-2.37 (m, 1H), 2.05-1.90 (m, 4H), 1.70-0.90 (m, 30H), 0.67 (s, 3H). LCMS Rt=1.226 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.30H.sub.44F.sub.3O.sub.4 [M+H—H.sub.2O].sup.+ 525, found 525.

Example 20. Synthesis of Compound 21

[0257] ##STR00044##

[0258] Synthesis of Compound 20-2. To a solution of Compound 18-1 (100 mg, 225 μmol) in DCM (3 mL) was added DMAP (2.22 mg, 67.5 μmol), TEA (68.3 mg, 675 μmol). DCC 17.5 mg, 675 μmol) and (S)-2-((tert-butoxycarbonyl)amino)propanoic acid (127 mg, 675 μmol). The mixture was stirred at 25° C. for 16 hours. The mixture was filtered, the filtrated was concentrated and purified by combi-flash (PE: EA=100%-95%) to give Compound 20-2 (120 mg, 87%) as an off-white solid, 1H NMR CDCl.sub.3 400 MHz δ 5.33-5.30 (m, 2H), 2.46-2.43 (m, 1H), 2.07-0.87 (m, 48H), 0.70 (s, 3H).

[0259] Synthesis of Compound 21. To a solution of Compound 20-2 (120 mg, 195 μmol) was added HCl/dioxane (4N, 1 mL). The mixture was stirred at 25° C. for 0.5 hour. To the mixture was added MTBE (3 mL) and stirred at 25° C. for 10 minutes. The mixture was filtered. The solid was washed with MTBE (3 mL), dried in vacuum to give Compound 21 (60 mg, 56%). .sup.1H NMR (400 MHz, MeOD) δ 5.52-5.51 (m, 1H), 5.32-5.31 (m, 1H), 4.30-4.26 (m, 1H), 2.46-2.43 (m, 1H), 2.05-1.00 (m, 36H), 0.76 (s, 3H). LCMS R.sub.t=0.940 min in 2 min chromatography, 30-90AB, MS ESI calcd. for C.sub.29H.sub.47F.sub.3NO.sub.3 [M+H].sup.+ 514, found 514.

Example 21. Synthesis of Compound 22

[0260] ##STR00045##

[0261] Step 1. To a mixture of Compound 18-1 (50 mg, 112 μmol) and DMAP (6.84 mg, 56.0 μmol) in pyridine (2 mL) was added pivalic anhydride (104 mg, 560 μmol) in one portion at 20° C. The mixture was stirred at 60° C. for 16 hrs. The resulting mixture was concentrated to give a residue, which was diluted with water (30 mL) and extracted with EtOAc (2×15 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give a crude product, which was purified by silica gel chromatography (PE/EtOAc=8/1) to give Compound 22 (14 mg, 24%) as an off-white solid. The reaction was conducted for a second time to give 30 mg of impure product. The 2 batches of product (30 mg) were combined and triturated with hexane (5 mL) to give Compound 22 (23 mg, 52%) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.33-5.19 (m, 2H), 2.45-2.38 (m, 1H), 2.03-1.91 (m, 3H), 1.86-0.80 (m, 40H), 0.60 (s, 3H). LCMS t.sub.R=1.575 min in 2 min chromatography, 30-90AB_ELSD. MS ESI calcd. for C.sub.31H.sub.48F.sub.3O.sub.2 [M+H—H.sub.2O].sup.+ 509, found 509.

Example 21. Synthesis of Compound 23

[0262] ##STR00046##

[0263] Step 1. To a solution of Compound 18-1 (100 mg, 0.225 mmol) in THF (5 mL) was added NaH (22.3 mg, 0.562 mmol, 60%) under N.sub.2 at 0° C. The mixture was stirred at 20° C. for 30 minutes. Acetyl chloride (35.3 mg, 0.45 mmol) was added. The reaction solution was stirred at 20° C. for 30 minutes. The mixture was quenched with Sat. NH.sub.4Cl (10 mL) and extracted with MTBE (3×5 mL). The combined organic phase was washed with brine, dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-8% of EtOAc in PE) to give Compound 23 (39 mg, 35%) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.30-5.29 (m, 2H), 2.48-2.38 (m, 1H), 2.14 (s, 3H), 2.05-1.95 (m, 3H), 1.94-1.56 (m, 6H), 1.54-1.47 (m, 8H), 1.45-0.92 (m, 17H), 0.59 (s, 3H). LCMS t.sub.R=1.383 min in 2 min chromatography, 30-90AB_ELSD. MS EST calcd. for C.sub.28H.sub.42F.sub.3O.sub.2 [M+H—H.sub.2O].sup.+ 467, found 467.

Example 22. Synthesis of Compound 24

[0264] ##STR00047##

[0265] Step 1. To a solution of Compound 22-1 (500 mg, 1.09 mmol; synthesized as described in Martinez et al., WO2014/160480, [00199]) in pyridine (3 mL) was added SO.sub.3—Py (519 mg, 3.27 mmol). The mixture was stirred at 50° C. for 1 h. The mixture was diluted with water (10 mL) and extracted with DCM (2×10 mL). The combined organic phase was washed with water (10 mL). To the organic phase was added dropwise NaOH solution (2 mL, 3% in water) with stirring and some white solid appeared. The solid was collected by filtration and washed with DCM (10 mL) and water (10 mL). The solid was purified by preparative-HPLC ((column: DuraShell 150*25 mm*5 um), gradient: 3 B (water (10 mM NH.sub.4HCO.sub.3), flow rate: 30 /min). After the purification was finished, to the combined eluent was added DCM (10 mL) and added dropwise NaOH solution (2 mL, 3% in water). Compound 24 (62 mg, 10%) was obtained by collection and lyophilization to give an off-white solid. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 5.36-5.35 (m, 1H), 2.88-2.85 (m, 1H), 2.35-2.31 (m, 1H), 2.21-2.01 (m, 2H), 2.00-1.75 (m, 4H), 1.70-1.43 (m, 10H), 1.41 (s, 3H), 1.38-1.30 (m, 1H), 1.27 (s, 3H), 1.22-1.10 (m, 5H), 1.09-1.02 (m, 4H), 1.00-0.92 (m, 4H), 0.75 (s, 3H). LCMS Rt=1.683 min in 2 min chromatography, 10-80CD_ELSD, MS ESI calcd. for C.sub.27H.sub.42F.sub.3O.sub.5S [M].sup.− 535, found 535.

Example 23. Synthesis of Compound 25

[0266] ##STR00048##

[0267] Step 1. To a solution of Compound 22-1 (500 mg, 1.09 mmol) in THF (10 mL) was added NaH (108 mg, 2.72 mmol, 60%) under N.sub.2 at 0° C. The mixture was stirred at 20° C. for 30 minutes. Then acetyl chloride (171 mg, 2.18 mmol) was added. The reaction solution was stirred at 20° C. for 1 h. The mixture was quenched with Sat. NH.sub.4Cl (10 mL) and extracted with EtOAc (3×10 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-30% of EtOAc in PE, 60 mins) to give Compound 25 (26 mg, 5%) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.35-5.25 (m, 1H), 2.50-2.40 (m, 1H), 2.30-2.10 (m, 1H), 2.06 (s, 3H), 2.00-1.60 (m, 12H), 1.53-1.35 (m, 7H), 1.30-0.90 (m, 17H), 0.67 (s, 3H). LCMS Rt=3.743 min in 7.0 min chromatography, 50-100 AB_E, MS ESI calcd. for C.sub.29H.sub.44F.sub.3O.sub.2 [M+H—H.sub.2O].sup.+ 481, found 481.

Example 24. Synthesis of Compound 26

[0268] ##STR00049##

[0269] Step 1. To a solution of Compound 24-1 (100 mg, 239 μmol; synthesized as described in Martinez et al., WO2014/160480, [00210]) in pyridine (2 mL) was added DMAP (30.5 mg, 478 μmol) and dihydrofuran-2,5-dione (119 mg, 1.19 mmol). The mixture was stirred at 60° C. for 40 hrs. The reaction mixture was washed by Sat.NH.sub.4Cl (3 ml) and extracted with ethyl acetate (2×2 ml). The combined organic layer was concentrated in vacuum. The residue was purified by silica gel chromatography (petroleum PE:EtOAc=10:1) to afford Compound 26 (35 mg, yield 28%) as an off-white solid. .sup.1H NMR CDCl.sub.3 400 MHz δ 5.35-5.30 (m, 1H), 2.64-2.61 (m, 5H), 2.58-2.56 (m, 1H), 2.01-0.86 (m, 42H), 0.67 (s, 3H). LCMS R.sub.t=1.341 min in 2 min chromatography, 30-90AB, MS ESI calcd. for C.sub.28H.sub.45 [M+H—H.sub.2O—HOOCCH.sub.2CH.sub.2COOH].sup.+ 381, found 381.

Example 25. Synthesis of Compounds 27 and 28

[0270] ##STR00050##

[0271] Step 1. To a solution of Compound 25-1 (900 mg, 2.23 mmol) in pyridine (20 mL) was added dihydrofuran-2,5-dione (1.11 g, 11.1 mmol) and DMAP (272 mg, 2.23 mmol). The mixture was stirred at 80° C. for 96 hrs. The reaction solution was quenched with sat.NH.sub.4Cl (25 mL) and extracted with EtOAc (3×10 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated. The reaction mixture was separated by combi-flash (0-10% of (MeOH: EtOAc, v: v=1:20) in DCM: PE, v: v=1:2) and the recycled starting material was reused in this experiment (this procedure was repeat for 4 times). The combined impure product was purified by preparative-HPLC (column: Boston Green ODS 150*30 5 u), gradient: 65-95% B (A=0.05% HCl/H.sub.2O, B=MeCN), flow rate: 75 mL/min) to give Compound 27 (40 mg, 34%) and Compound 28 (55 mg, 4%) as off-white solids.

[0272] 27: .sup.1H NMR (400 MHz, CDCl.sub.3) 5.30-5.29 (m, 1H), 2.65-2.55 (m, 4H), 2.45-2.35 (m, 1H), 2.05-1.92 (m, 3H), 1.90-1.76 (m, 3H), 1.75-1.53 (m, 5H), 1.52-1.46 (m, 3H), 1.45-1.40 (m, 7H), 1.39-1.30 (m, 2H), 1.29-1.20 (m, 1H), 1.19-1.15 (m, 2H), 1.14-1.10 (m, 4H), 1.09-1.02 (m, 2H), 1.01-0.94 (m, 4H), 0.93-0.86 (m, 5H), 0.67 (s, 3H). LCMS Rt=1.268 min in 2.0 min chromatography, 30-90AB_ELSD, MS ESI calcd. for C.sub.27H.sub.43 [M+H—H.sub.2—(CH.sub.2COOH).sub.2].sup.+ 367, found 367. HRMS MS ESI calcd. for C.sub.31H.sub.49O.sub.5 [M−H].sup.− 501.3585, found 501.3575. 28: .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.30-5.29 (m, 1H), 2.66-2.52 (m, 5H), 2.38-2.32 (m, 1H), 2.08-1.77 (m, 6H), 1.76-1.71 (m, 1H), 1.70-1.52 (m, 3H), 1.51-1.40 (m, 10H), 1.39 (s, 3H), 1.38-1.18 (m, 6H), 1.17-1.11 (m, 4H), 1.10 (s, 3H), 1.09-0.95 (m, 1H), 0.94-0.88 (m, 3H), 0.68 (s, 3H). LCMS Rt=1.280 min in 2.0 min chromatography, 30-90AB_ELSD, MS ESI calcd. for C.sub.27H.sub.43 [M+H—H.sub.2O—(CH.sub.2COOH).sub.2].sup.+ 367, found 367. HRMS MS ESI calcd. for C.sub.31H.sub.49O.sub.5 [M−H].sup.− 501.3585, found 501.3597.

Example 26. Synthesis of Compound 29

[0273] ##STR00051##

[0274] Step 1. To a solution of Compound 29-1 (1.5 g, 3.72 mmol) in DCM (30 mL) was added DMAP (226 mg, 1.86 mmol) and Py (8.65 g, 111 mmol), followed by adding bimethyl phosphorochloridate (2.68 g, 18.6 mmol) dropwise under N.sub.2. The reaction was stirred at 20° C. for 2 hrs. The mixture was quenched with sat.NaHCO.sub.3 (100 mL) and extracted with DCM (3×40 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-50% of EtOAc in PE/DCM (v/v=2/1)) to give Compound 29-2 (620 mg, 27%) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.37 (m, 1H), 4.35-4.15 (m, 2H), 3.77 (s, 6H), 3.74 (s, 6H), 2.45-2.35 (m, 2H), 2.05-1.87 (m, 4H), 1.86-1.78 (m, 2H), 1.77-1.58 (m, 2H), 1.56-1.33 (m, 7H), 1.30-1.15 (m, 4H), 1.14-1.04 (m, 3H), 1.02 (s, 3H), 1.01-0.88 (m, 11H), 0.67 (s, 3H).

[0275] Step 2. To a solution of Compound 29-2 (200 mg, 0.323 mmol) in DCM (5 mL) was added TMSBr (246 mg, 1.61 mmol) under N.sub.2. The mixture was stirred at 20° C. for 16 hrs. The reaction mixture was adjusted to pH=9 with aq. NaOH (1.9 mL, 1 M) and the solid precipitated. The white solid was filtered and washed with CH.sub.2C.sub.2 (2 mL), water (2 mL), dried and lyophilized to give Compound 29 (68 mg, 37%) as an off-white solid. .sup.1H NMR (400 MHz, MeOD) δ 5.40-5.37 (m, 1H), 4.10-3.90 (m, 2H), 2.48-2.30 (m, 2H), 2.08-1.85 (m, 7H), 1.70-1.58 (m, 10H), 1.35-1.25 (m, 1H), 1.20-1.04 (m, 5H), 1.02 (s, 3H), 1.00-0.96 (m, 7H), 0.95-0.88 (m, 3H), 0.71 (s, 3H). LCMS t.sub.R=0.207 min in 3 min chromatography, 10-80CD_ELSD. MS ESI calcd. for C.sub.27H.sub.47O.sub.8P.sub.2 [M-4Na+4H—H].sup.− 561, found 561.

Example 27. Synthesis of Compound 30

[0276] ##STR00052##

[0277] Step 1. To a solution of 29-1 (1.5 g, 3.72 mol) in DCM (30 L) was added DMAP (226 mg, 1.86 mmol) and Py (8.65 g, 111 mol), followed by adding bimethyl phosphorochloridate (2.68 g, 18.6 mmol) dropwise under N.sub.2. The reaction was stirred at 20° C. for 2 hrs. The mixture was quenched with sat.NaHCO.sub.3 (100 mL) and extracted with DCM (3×40 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-50% of EtOAc in PE/DCM (v/v=2/1)) to give 27-1 (700 mg, 37%) as an off-white solid.

[0278] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.37 (m, 1H), 4.28-4.15 (m, 1H), 3.80-3.70 (m, 6H), 3.35-3.25 (m, 1H), 2.45-2.40 (m, 2H), 2.02-1.90 (m, 3H), 1.89-1.76 (m, 2H), 1.75-1.58 (m, 2H), 1.56-1.33 (m, 8H), 1.30-1.15 (m, 7H), 1.14-0.96 (m, 5H), 0.95-0.88 (m, 10H), 0.67 (s, 3H).

[0279] Step 2. To a solution of 27-1 (200 mg, 0.391 mmol) in DCM (5 mL) was added TMSBr (298 mg, 1.95 mmol). The mixture was stirred at 20° C. for 16 hrs. Another TMSBr (596 mg, 3.9 mmol) was added at 0° C. The mixture was stirred at 20° C. for 16 hrs. The mixture was adjusted with NaOH (1M in H.sub.2O) to pH=9 and the solid was precipitated. The white solid was filtered and washed with CH.sub.2Cl.sub.2 (2 mL), water (2 mL), dissolved in MeOH (6 mL) and concentrated to give Compound 30 (16 mg, 8%) as an off-white solid. .sup.1H NMR (400 MHz, MeOD) δ 5.42-5.29 (m, 1H), 4.04-3.82 (m, 1H), 3.25-3.15 (m, 1H), 2.63-2.41 (m, 1H), 2.35-2.11 (m, 1H), 2.10-1.80 (m, 4H), 1.79-1.37 (m, 10H), 1.37-1.27 (m, 2H), 1.26-1.05 (m, 6H), 1.02 (s, 3H), 0.99-0.77 (m, 13H), 0.72 (s, 3H). LCMS t.sub.R=1.311 min in 3 min chromatography, 10-80CD_ELSD, MS ESI calcd. for C.sub.27H.sub.46O.sub.5P [M-2Na+2H—H].sup.− 481, found 481.

Example 28. Alternative Synthesis of Compound 6

[0280] ##STR00053##

[0281] Step 1. To a solution of 29-1 (1.4 g, 3.47 mmol) in DCM (25 mL) was added imidazole (471 mg, 6.94 mmol). A solution of TBSCl (1.39 g, 10.4 mmol) in DCM (5 mL) was added dropwise under N.sub.2. The reaction was stirred at 20° C. for 16 hrs. The mixture was quenched with water (30 mL) and extracted with DCM (2×15 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-5% of EtOAc in PE) to give 28-1 (1.5 g, 84%) as an off-white solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.33-5.30 (m, 1H), 3.52-3.43 (m, 1H), 3.34-3.27 (m, 1H), 2.31-2.13 (m, 2H), 2.04-1.92 (m, 2H), 1.90-1.76 (m, 2H), 1.75-1.57 (m, 4H), 1.54-1.37 (m, 7H), 1.34-1.19 (m, 4H), 1.18-1.01 (m, 5H), 0.99 (s, 3H), 0.96-0.89 (m, 9H), 0.88-0.85 (m, 10H), 0.67 (s, 3H), 0.05 (s, 6H).

[0282] Step 2. To a solution of 28-1 (900 g, 1.74 mmol) in THF (30 mL) was added n-BuLi (1.04 mL, 2.61 mmol, 2.5M in hexane) dropwise at −70° C. under N.sub.2. The reaction solution was stirred at −70° C. for 30 minutes. Then dimethyl phosphorochloridate (502 mg, 3.48 mmol) was added dropwise. After addition, the mixture was stirred at 20° C. for 2 hrs. The reaction solution was quenched with sat.NH.sub.4Cl (50 mL) and extracted with DCM (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-20% of EtOAc in PE) to give 6-3A (540 mg, 50%) as colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.33-5.29 (m, 1H), 4.23-4.15 (m, 1H), 3.81-3.71 (m, 6H), 3.53-3.43 (m, 1H), 2.32-2.22 (m, 1H), 2.18-1.64 (m, 9H), 1.60-1.33 (m, 11H), 1.31-1.06 (m, 5H), 0.99 (s, 3H), 0.97-0.90 (m, 10H), 0.89-0.85 (m, 8H), 0.67 (s, 3H), 0.05 (s, 6H).

[0283] Step 3. To a solution of 6-3A (180 mg, 0.288 mmol) in DCM (5 mL) was added TMSBr (220 mg, 1.44 mmol). The mixture was stirred at 20° C. for 16 hrs. The mixture was adjusted with NaOH (1M in H.sub.2O) to pH=9 and the solid was precipitated, which was filtered and washed with CH.sub.2Cl.sub.2 (2 mL), water (2 mL). The filter cake was dissolved in MeOH (6 mL) and concentrated to give Compound 6 (35 mg, 25%) as an off-white solid. .sup.1H NMR (400 MHz, MeOD) δ 5.37-5.31 (m, 1H), 4.01-3.89 (m, 1H), 3.45-3.35 (m, 1H), 2.29-2.14 (m, 2H), 2.09-1.84 (m, 5H), 1.82-1.72 (m, 1H), 1.66-1.38 (m, 10H), 1.37-1.26 (m, 1H), 1.24-1.04 (m, 5H), 1.03-1.00 (m, 4H), 0.99-0.93 (m, 7H), 0.92-0.85 (m, 4H), 0.72 (s, 3H). LCMS t.sub.R=1.370 min in 3 min chromatography, 10-80CD_ELSD. MS ESI calcd. for C.sub.27H.sub.46O.sub.5P [M-2Na+2H—H].sup.− 481, found 481.

Materials and Methods

[0284] The compounds provided herein can be prepared from readily available starting materials using the following general methods and procedures, for example, as described in WO 2013/036835 and WO 2014/160480. It will be appreciated that where typical or preferred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimization.

[0285] Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group as well as suitable conditions for protection and deprotection are well known in the art. For example, numerous protecting groups, and their introduction and removal, are described in T. W. Greene and P. G. M. Wuts, Protecting Groups in Organic Synthesis, Second Edition. Wiley, New York, 1991, and references cited therein.

[0286] The compounds provided herein may be isolated and purified by known standard procedures. Such procedures include (but are not limited to) recrystallization, column chromatography, HPLC, or supercritical fluid chromatography (SFC). The compounds provided herein may be prepared from known or commercially available starting materials and reagents by one skilled in the art of organic synthesis. Exemplary chiral columns available for use in the separation/purification of the enantiomers/diastereomers provided herein include, but are not limited to, CHIRALPAK® AD-10. CHIRALCEL® OB, CHIRALCEL® OB-H. CHIRALCEL® OD. CHIRALCEL® OD-H, CHIRALCEL® OF. CHIRALCEL® OG, CHIRALCEL® OJ and CHIRALCEL® OK.

[0287] .sup.1H-NMR reported herein (e.g., for the region between δ (ppm) of about 0.5 to about 4 ppm) will be understood to be an exemplary interpretation of the NMR spectrum (e.g., exemplary peak integrations) of a compound. Exemplary general method for preparative HPLC: Column: Waters RBridge prep 10 μm C18. 19*250 mm. Mobile phase: acetonitrile, water (NH.sub.4HCO.sub.3) (30 L water, 24 g NH.sub.4HCO.sub.3, 30 mL NH.sub.3.H.sub.2O). Flow rate: 25 mL/min

[0288] Exemplary general method for analytical HPLC: Mobile phase: A: water (10 mM NH.sub.4HCO.sub.3). B: acetonitrile Gradient: 5%-95% B in 1.6 or 2 min Flow rate: 1.8 or 2 mL/min; Column: XBridge C18, 4.6*50 mm, 3.5 μm at 45 C.

OTHER EMBODIMENTS

[0289] 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.

[0290] 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.

[0291] 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.

[0292] 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.