OXYSTEROLS AND METHODS OF USE THEREOF

Abstract

Compounds are provided according to Formula (I):

##STR00001##

and pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof; wherein R.sup.2, R.sup.3, R.sup.4, R.sup.5, and R.sup.6 are as defined herein. Compounds of the present invention are contemplated useful for the prevention and treatment of a variety of conditions.

Claims

1. A compound of Formula (I-59): ##STR00639## or a pharmaceutically acceptable salt thereof, wherein: each of R.sup.2 and R.sup.3 is independently hydrogen, substituted or unsubstituted C.sub.1-C.sub.6 alkyl, substituted or unsubstituted carbocyclyl, or substituted or unsubstituted heterocyclyl, or R.sup.2 and R.sup.3, together with the carbon atom to which they are attached form a substituted or unsubstituted 3-8 membered ring; each of R.sup.4 and R.sup.5 is independently hydrogen, halo, or —OR.sup.C, wherein R.sup.C is hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl, or R.sup.4 and R.sup.5, together with the carbon atom to which they are attached form an oxo group; R.sup.6 is absent or hydrogen; and represents a single or double bond, wherein when one of is a double bond, the other is a single bond; when both of are single bonds, then R.sup.6 is hydrogen; and when one of is a double bond, R.sup.6 is absent; provided that the following compounds are excluded: ##STR00640##

2. The compound of claim 1, wherein R.sup.2 is hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl.

3. The compound of claim 1, wherein R.sup.2 is C.sub.1-C.sub.6 haloalkyl.

4. The compound of claim 1, wherein each of R.sup.2 and R.sup.3 is independently substituted C.sub.1-C.sub.6 alkyl or hydrogen.

5. The compound of claim 1, wherein each of R.sup.2 and R.sup.3 is independently unsubstituted C.sub.1-C.sub.6 alkyl or hydrogen.

6. The compound of claim 1, wherein each of R.sup.2 and R.sup.3 is independently C.sub.1-C.sub.6 haloalkyl or hydrogen.

7. The compound of claim 1, wherein each of R.sup.2 and R.sup.3 is independently hydrogen, carbocyclyl, or heterocyclyl.

8. The compound of claim 1, wherein each of R.sup.2 and R.sup.3 is independently C.sub.2-C.sub.6 alkyl or hydrogen.

9. The compound of claim 1, wherein each of R.sup.2 and R.sup.3 is independently isopropyl or tert-butyl or hydrogen.

10. The compound of claim 1, wherein at least one of R.sup.2 and R.sup.3 is C.sub.3-C.sub.6 alkyl, carbocyclyl, or 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.

11. The compound of claim 1, wherein R.sup.2 is isopropyl or tert-butyl and R.sup.3 is methyl or hydrogen.

12. The compound of claim 1, wherein R.sup.2 is substituted isopropyl or substituted tert-butyl and R.sup.3 is unsubstituted methyl or hydrogen.

13. The compound of claim 1, wherein R.sup.2 is unsubstituted isopropyl or unsubstituted tert-butyl and R.sup.3 is unsubstituted methyl or hydrogen.

14. The compound of claim 1, wherein R.sup.2 is tert-butyl and R.sup.3 is hydrogen.

15. The compound of claim 1, wherein R.sup.2 is substituted tert-butyl and R.sup.3 is hydrogen.

16. The compound of claim 1, wherein R.sup.2 is unsubstituted tert-butyl and R.sup.3 is hydrogen.

17. The compound of claim 1, wherein R.sup.2 is trifluoromethyl and R.sup.3 is hydrogen.

18. The compound of claim 1, wherein R.sup.2 is trifluoromethyl and R.sup.3 is methyl.

19. The compound of claim 1, wherein R.sup.2 is trifluoromethyl and R.sup.3 is substituted methyl.

20. The compound of claim 1, wherein R.sup.2 is trifluoromethyl and R.sup.3 is unsubstituted methyl.

21. The compound of claim 1, wherein R.sup.2 is methyl and R.sup.3 is hydrogen.

22. The compound of claim 1, wherein R.sup.2 is substituted methyl and R.sup.3 is hydrogen.

23. The compound of claim 1, wherein R.sup.2 is unsubstituted methyl and R.sup.3 is hydrogen.

24. The compound of claim 1, wherein R.sup.4 is —OH or halo.

25. The compound of claim 1, wherein R.sup.4 and R.sup.5, together with the carbon atom to which they are attached form an oxo group.

26. The compound of claim 1, wherein R.sup.4 is hydrogen and R.sup.5 is halo.

27. The compound of claim 1, wherein R.sup.4 and R.sup.5 are halo.

28. The compound of claim 1, wherein R.sup.4 and R.sup.5 are hydrogen.

29. The compound of claim 1, wherein R.sup.2 and R.sup.3, together with the carbon atom to which they are attached form a 5-membered ring.

30. The compound of claim 1, wherein R.sup.2 is C.sub.2-C.sub.6 alkyl and R.sup.3 is C.sub.1-C.sub.6 alkyl.

31. The compound of claim 1, wherein R.sup.2 is unsubstituted C.sub.2-C.sub.6 alkyl and R.sup.3 is unsubstituted C.sub.1-C.sub.6 alkyl.

32. The compound of claim 1, wherein R.sup.2 and R.sup.3, together with the carbon atom to which they are attached form a 6-membered ring.

33. The compound of claim 1, wherein R.sup.2 is carbocyclyl or heterocyclyl and R.sup.3 is hydrogen.

34. The compound of claim 1, wherein R.sup.2 and R.sup.3 are hydrogen.

35. The compound of claim 1, wherein R.sup.2 is isopropyl and R.sup.3 is hydrogen.

36. The compound of claim 1, wherein R.sup.2 is substituted isopropyl and R.sup.3 is hydrogen.

37. The compound of claim 1, wherein R.sup.2 is substituted isopropyl and R.sup.3 is hydrogen.

38. The compound of claim 1, wherein R.sup.2 and R.sup.3, together with the carbon atom to which they are attached form a 3-8 membered carbocyclic or heterocyclic ring.

39. The compound of claim 33, wherein the carbocyclic or heterocyclic ring is substituted with 1 or 2 halo or alkyl groups.

40. The compound of claim 1, wherein R.sup.2 is cyclobutyl and R.sup.3 is hydrogen.

41. The compound of claim 1, wherein R.sup.2 is tetrahydropyranyl and R.sup.3 is hydrogen.

42. The compound of claim 1, wherein R.sup.2 is substituted cyclobutyl and R.sup.3 is hydrogen.

43. The compound of claim 1, wherein R.sup.2 is substituted tetrahydropyranyl and R.sup.3 is hydrogen.

44. The compound of claim 1, wherein R.sup.2 is unsubstituted cyclobutyl and R.sup.3 is hydrogen.

45. The compound of claim 1, wherein R.sup.2 is unsubstituted tetrahydropyranyl and R.sup.3 is hydrogen.

46. The compound of claim 1, wherein the compound of Formula (I-59) is selected from a compound of Formula (I-A59), (I-B59), or (I-C59): ##STR00641##

47. The compound of claim 1, wherein the compound of Formula (I-59) is selected from a compound of Formula (I-B59): ##STR00642##

48. The compound of claim 1, wherein the compound of Formula (I-59) is selected from a compound of Formula (I-C59): ##STR00643##

49. The compound of claim 1, wherein at least one of R.sup.2 and R.sup.3 is hydrogen, C.sub.1-C.sub.6 alkyl, carbocyclyl, or 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.

50. The compound of claim 1, wherein the compound of Formula (I) is selected from a compound of Formula (I-D59): ##STR00644##

51. The compound of claim 1, wherein the compound of Formula (I-59) is selected from a compound of Formula (I-E59): ##STR00645##

52. The compound of claim 1, wherein the compound of Formula (I-59) is selected from a compound of Formula (I-D-i59) or (I-D-ii59): ##STR00646##

53. The compound of claim 1, wherein the compound of Formula (I-59) is selected from a compound of Formula (I-E-i59) or (I-E-ii59): ##STR00647##

54. The compound of claim 1, wherein the compound is: ##STR00648## ##STR00649## ##STR00650##

55. A pharmaceutical composition comprising a compound of claim 1, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

56. A method for treating or preventing a disorder described herein, comprising administering to a subject in need thereof an effective amount of a compound of claim 1, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof.

57. The method according to claim 56, wherein the disorder is a gastrointestinal (GI) disorder, constipation, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, structural disorders affecting the GI, anal disorders, hemorrhoids, internal hemorrhoids, external hemorrhoids, anal fissures, perianal abscesses, anal fistula, colon polyps, cancer, or colitis.

58. The method according to claim 56, wherein the disorder is inflammatory bowel disease.

59. The method according to claim 56, wherein the disorder is cancer, diabetes, or a sterol synthesis disorder.

60. The method according to claim 56, wherein the disorder is a metabolic disorder.

61. The method according to claim 56, wherein the disorder is an autoimmune disorder.

62. The method according to claim 56, wherein the disorder is rheumatoid arthritis, juvenile idiopathic arthritis, ankylosing spondylitis, psoriatic arthritis, Crohn's disease, ulcerative colitis, and plaque psoriasis.

63. A method for treating or preventing a CNS-related condition comprising administering to a subject in need thereof an effective amount of a compound of claim 1, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof.

64. The method according to claim 63, wherein the CNS-related condition is an adjustment disorder, anxiety disorder (including obsessive-compulsive disorder, posttraumatic stress disorder, and social phobia), cognitive disorder (including Alzheimer's disease and other forms of dementia), dissociative disorder, eating disorder, mood disorder (including depression (e.g., postpartum depression), bipolar disorder, dysthymic disorder, suicidality), schizophrenia or other psychotic disorder (including schizoaffective disorder), sleep disorder (including insomnia), substance-related disorder, personality disorder (including obsessive-compulsive personality disorder), autism spectrum disorders (including those involving mutations to the Shank group of proteins, (Shank3)), neurodevelopmental disorder (including Rett syndrome, Tuberous Sclerosis complex), multiple sclerosis, sterol synthesis disorders, pain (including acute and chronic pain), encephalopathy secondary to a medical condition (including hepatic encephalopathy and anti-NMDA receptor encephalitis), seizure disorder (including status epilepticus and monogenic forms of epilepsy such as Dravet's disease), stroke, traumatic brain injury, movement disorder (including Huntington's disease and Parkinson's disease), vision impairment, hearing loss, or tinnitus.

65. The method according to claim 63, wherein the disorder is a sterol synthesis disorder.

66. A compound of Formula (I-66): ##STR00651## or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 is substituted or unsubstituted C.sub.1-C.sub.6 alkyl; R.sup.2 is substituted or unsubstituted aralkyl, substituted or unsubstituted heteroaralkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R.sup.3 is hydrogen, substituted or unsubstituted C.sub.1-C.sub.6 alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, aryl, or substituted or unsubstituted heteroaryl; each of R.sup.4 and R.sup.5 is independently hydrogen, halo, or —OR.sup.C, wherein R.sup.C is hydrogen or unsubstituted or substituted C.sub.1-C.sub.3 alkyl, or R.sup.4 and R.sup.5, together with the carbon atom to which they are attached form an oxo group; R.sup.6 is absent or hydrogen; and represents a single or double bond, wherein when one of is a double bond, the other is a single bond; when both of are single bonds, then R.sup.6 is hydrogen; and when one of is a double bond, R.sup.6 is absent.

67. The compound of claim 66, wherein R.sup.1 is substituted C.sub.1-C.sub.6 alkyl.

68. The compound of claim 66, wherein R.sup.1 is unsubstituted C.sub.1-C.sub.6 alkyl.

69. The compound of claim 66, wherein R.sup.1 is —CH.sub.3, —CF.sub.3, or —CH.sub.2CH.sub.3.

70. The compound of claim 66, wherein R.sup.1 is —CH.sub.2OR.sup.A, wherein R.sup.A is substituted or unsubstituted C.sub.1-C.sub.6 alkyl.

71. The compound of claim 66, wherein R.sup.2 is substituted or unsubstituted aryl, substituted or unsubstituted phenyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted pyridyl, or substituted or unsubstituted aralkyl.

72. The compound of claim 66, wherein R.sup.2 is substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl, or substituted or unsubstituted benzyl.

73. The compound of claim 66, wherein R.sup.3 is hydrogen or C.sub.1-C.sub.6 alkyl.

74. The compound of claim 66, wherein R.sup.3 is hydrogen, unsubstituted C.sub.1-C.sub.6 alkyl, or haloalkyl.

75. The compound of claim 66, wherein R.sup.4 is —OH or halo.

76. The compound of claim 66, wherein R.sup.4 and R.sup.5, together with the carbon atom to which they are attached form an oxo group.

77. The compound of claim 66, wherein R.sup.4 is hydrogen and R.sup.5 is halo.

78. The compound of claim 66, wherein R.sup.4 and R.sup.5 are halo.

79. The compound of claim 66, wherein R.sup.4 and R.sup.5 are hydrogen.

80. The compound of claim 66, wherein R.sup.2 is substituted or unsubstituted aryl, substituted or unsubstituted phenyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted pyridyl, substituted or unsubstituted aralkyl, substituted or unsubstituted benzyl, or heteroaralkyl and R.sup.3 is hydrogen or unsubstituted C.sub.1-C.sub.6 alkyl, or C.sub.1-C.sub.6 haloalkyl.

81. The compound of claim 66, wherein R.sup.2 is substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted pyridyl, substituted or unsubstituted aralkyl, substituted or unsubstituted benzyl, or heteroaralkyl and R.sup.3 is hydrogen, —CH.sub.3, or —CF.sub.3.

82. The compound of claim 66, wherein R.sup.1 is C.sub.1-C.sub.6 alkyl, R.sup.2 is substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted pyridyl, substituted or unsubstituted aralkyl, substituted or unsubstituted benzyl, or heteroaralkyl, and R.sup.3 is hydrogen, —CH.sub.3, or —CF.sub.3.

83. The compound of claim 66, wherein R.sup.1 is —CH.sub.3 or —CH.sub.2CH.sub.3, R.sup.2 is unsubstituted phenyl, unsubstituted pyridyl, or unsubstituted benzyl, and R.sup.3 is hydrogen, —CH.sub.3, or —CF.sub.3.

84. The compound of claim 66, wherein the compound of Formula (I-66) is selected from a compound of Formula (I-A66), (I-B66), or (I-C66): ##STR00652##

85. The compound of claim 66, wherein the compound of Formula (I-66) is selected from a compound of Formula (I-A66): ##STR00653##

86. The compound of claim 66, wherein the compound is: ##STR00654## ##STR00655##

87. A pharmaceutical composition comprising a compound of claim 66, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

88. A method for treating or preventing a disorder described herein, comprising administering to a subject in need thereof an effective amount of a compound of claim 66, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof.

89. The method according to claim 88, wherein the disorder is a gastrointestinal (GI) disorder constipation, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD) ulcerative colitis, Crohn's disease, structural disorders affecting the GI, anal disorders, hemorrhoids, internal hemorrhoids, external hemorrhoids, anal fissures, perianal abscesses, anal fistula, colon polyps, cancer, or colitis.

90. The method according to claim 88, wherein the disorder is inflammatory bowel disease.

91. The method according to claim 88, wherein the disorder is cancer, diabetes, or a sterol synthesis disorder.

92. The method according to claim 88, wherein the disorder is a metabolic disorder.

93. The method according to claim 88, wherein the disorder is an autoimmune disorder.

93. The method according to claim 88, wherein the disorder is rheumatoid arthritis, juvenile idiopathic arthritis, ankylosing spondylitis, psoriatic arthritis, Crohn's disease, ulcerative colitis, and plaque psoriasis.

94. A method for treating or preventing a CNS-related condition comprising administering to a subject in need thereof an effective amount of a compound of claim 66, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof.

95. The method according to claim 94, wherein the CNS-related condition is an adjustment disorder, anxiety disorder (including obsessive-compulsive disorder, posttraumatic stress disorder, and social phobia), cognitive disorder (including Alzheimer's disease and other forms of dementia), dissociative disorder, eating disorder, mood disorder (including depression (e.g., postpartum depression), bipolar disorder, dysthymic disorder, suicidality), schizophrenia or other psychotic disorder (including schizoaffective disorder), sleep disorder (including insomnia), substance-related disorder, personality disorder (including obsessive-compulsive personality disorder), autism spectrum disorders (including those involving mutations to the Shank group of proteins (e.g., Shank3)), neurodevelopmental disorder (including Rett syndrome, Tuberous Sclerosis complex), multiple sclerosis, sterol synthesis disorders, pain (including acute and chronic pain), encephalopathy secondary to a medical condition (including hepatic encephalopathy and anti-NMDA receptor encephalitis), seizure disorder (including status epilepticus and monogenic forms of epilepsy such as Dravet's disease), stroke, traumatic brain injury, movement disorder (including Huntington's disease and Parkinson's disease), vision impairment, hearing loss, or tinnitus.

96. The method according to claim 88, wherein the disorder is a sterol synthesis disorder.

97. A compound of Formula (I-61): ##STR00656## or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 is hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl; each of R.sup.2 and R.sup.3 is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted carbocyclyl, or substituted or unsubstituted heterocyclyl or R.sup.2 and R.sup.3, together with the carbon atom to which they are attached for a 3-8 substituted or unsubstituted membered ring; each of R.sup.4 and R.sup.5 is independently hydrogen, halo, or —OR.sup.C, wherein R.sup.C is hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl, or R.sup.4 and R.sup.5, together with the carbon atom to which they are attached form an oxo group; R.sup.6 is absent or hydrogen; and represents a single or double bond, wherein when one of is a double bond, the other is a single bond; when both of are single bonds, then R.sup.6 is hydrogen; and when one of is a double bond, R.sup.6 is absent; provided that the following compounds are excluded: ##STR00657## ##STR00658## ##STR00659##

98. The compound of claim 97, wherein R.sup.1 is C.sub.1-C.sub.6 alkyl or hydrogen.

99. The compound of claim 97, wherein R.sup.1 is C.sub.2-C.sub.6 alkyl or hydrogen.

100. The compound of claim 97, wherein R.sup.1 is substituted or unsubstituted substituted or unsubstituted C.sub.3-C.sub.6 alkyl or hydrogen.

101. The compound of claim 97, wherein R.sup.1 is substituted or unsubstituted methyl or substituted or unsubstituted ethyl.

102. The compound of claim 97, wherein R.sup.1 is substituted or unsubstituted methyl or substituted or unsubstituted ethyl.

103. The compound of claim 97, wherein R.sup.1 is trifluoromethyl.

104. The compound of claim 97, wherein R.sup.1 is —CH.sub.2OR.sup.A, wherein R.sup.A is C.sub.1-C.sub.6 alkyl.

105. The compound of claim 97, wherein R.sup.2 is hydrogen or C.sub.2-C.sub.6 alkyl.

106. The compound of claim 97, wherein R.sup.2 is hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl.

107. The compound of claim 97, wherein R.sup.2 is hydrogen.

108. The compound of claim 97, wherein R.sup.2 is substituted or unsubstituted isopropyl.

109. The compound of claim 97, wherein R.sup.2 is substituted or unsubstituted isopropyl.

110. The compound of claim 97, wherein R.sup.2 is C.sub.1-C.sub.6 haloalkyl.

111. The compound of claim 97, wherein each of R.sup.2 and R.sup.3 is independently C.sub.1-C.sub.6 alkyl or hydrogen.

112. The compound of claim 97, wherein each of R.sup.2 and R.sup.3 is independently substituted or unsubstituted C.sub.1-C.sub.6 alkyl or hydrogen.

113. The compound of claim 97, wherein R.sup.2 and R.sup.3, together with the carbon atom to which they are attached for a 3-8 membered ring.

114. The compound of claim 97, wherein each of R.sup.2 and R.sup.3 is independently hydrogen or C.sub.1-C.sub.6 alkyl.

115. The compound of claim 97, wherein each of R.sup.2 and R.sup.3 is independently hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl.

116. The compound of claim 97, wherein each of R.sup.2 and R.sup.3 is independently hydrogen, C.sub.3-C.sub.6 alkyl, or isopropyl.

117. The compound of claim 97, wherein each of R.sup.2 and R.sup.3 is independently hydrogen, substituted or unsubstituted C.sub.3-C.sub.6 alkyl, or substituted or unsubstituted isopropyl.

118. The compound of claim 97, wherein R.sup.4 is —OH or halo.

119. The compound of claim 97, wherein R.sup.4 and R.sup.5, together with the carbon atom to which they are attached form an oxo group.

120. The compound of claim 97, wherein R.sup.4 is hydrogen and R.sup.5 is halo.

121. The compound of claim 97, wherein R.sup.4 and R.sup.5 are halo.

122. The compound of claim 97, wherein R.sup.4 and R.sup.5 are fluorine.

123. The compound of claim 97, wherein R.sup.4 and R.sup.5 are hydrogen.

124. The compound of claim 97, wherein R.sup.2 and R.sup.3 are hydrogen.

125. The compound of claim 97, wherein R.sup.2 is C.sub.1-C.sub.6 alkyl and R.sup.3 is C.sub.2-C.sub.6 alkyl.

126. The compound of claim 97, wherein R.sup.2 is substituted or unsubstituted C.sub.1-C.sub.6 alkyl and R.sup.3 is substituted or unsubstituted C.sub.2-C.sub.6 alkyl.

127. The compound of claim 97, wherein R.sup.1 is substituted or unsubstituted ethyl and R.sup.2 and R.sup.3 are substituted or unsubstituted methyl.

128. The compound of claim 97, wherein R.sup.1 is substituted or unsubstituted ethyl and R.sup.2 and R.sup.3 are substituted or unsubstituted methyl.

129. The compound of claim 97, wherein R.sup.1 is ethyl, R.sup.2 is isopropyl, and R.sup.3 is hydrogen.

130. The compound of claim 97, wherein R.sup.1 is substituted or unsubstituted ethyl, R.sup.2 is substituted or unsubstituted isopropyl, and R.sup.3 is hydrogen.

131. The compound of claim 97, wherein R.sup.1 is ethyl, R.sup.2 is isopropyl, and R.sup.3 is methyl.

132. The compound of claim 97, wherein R.sup.1 is substituted or unsubstituted ethyl, R.sup.2 is substituted or unsubstituted isopropyl, and R.sup.3 is substituted or unsubstituted methyl.

133. The compound of claim 97, wherein the compound of Formula (I-61) is a compound of Formula (I-A61), (I-B61), or (I-C61): ##STR00660##

134. The compound of claim 97, wherein the compound of Formula (I-61) is selected from a compound of Formula (I-C61): ##STR00661##

135. The compound of claim 97, wherein the compound of Formula (I-61) is selected from a compound of Formula (I-A61): ##STR00662##

136. The compound of claim 97, wherein the compound of Formula (I-61) is selected from a compound of Formula (I-C-i61) or (I-C-ii61): ##STR00663##

137. The compound of claim 97, wherein the compound is: ##STR00664## ##STR00665##

138. A pharmaceutical composition comprising a compound of claim 97, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

139. A method for treating or preventing a disorder described herein, comprising administering to a subject in need thereof an effective amount of a compound of claim 97, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof.

140. The method according to claim 139, wherein the disorder is a gastrointestinal (GI) disorder, constipation, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, structural disorders affecting the GI, anal disorders, hemorrhoids, internal hemorrhoids, external hemorrhoids, anal fissures, perianal abscesses, anal fistula, colon polyps, cancer, or colitis.

141. The method according to claim 139, wherein the disorder is inflammatory bowel disease.

142. The method according to claim 139, wherein the disorder is cancer, diabetes, or a sterol synthesis disorder.

143. The method according to claim 139, wherein the disorder is a metabolic disorder.

144. The method according to claim 139, wherein the disorder is an autoimmune disorder.

145. The method according to claim 139, wherein the disorder is rheumatoid arthritis, juvenile idiopathic arthritis, ankylosing spondylitis, psoriatic arthritis, Crohn's disease, ulcerative colitis, and plaque psoriasis.

146. A method for treating or preventing a CNS-related condition comprising administering to a subject in need thereof an effective amount of a compound of claim 97, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof.

147. The method according to claim 139, wherein the CNS-related condition is an adjustment disorder, anxiety disorder (including obsessive-compulsive disorder, posttraumatic stress disorder, and social phobia), cognitive disorder (including Alzheimer's disease and other forms of dementia), dissociative disorder, eating disorder, mood disorder (including depression (e.g., postpartum depression), bipolar disorder, dysthymic disorder, suicidality), schizophrenia or other psychotic disorder (including schizoaffective disorder), sleep disorder (including insomnia), substance-related disorder, personality disorder (including obsessive-compulsive personality disorder), autism spectrum disorders (including those involving mutations to the Shank group of proteins (including Shank3)), neurodevelopmental disorder (including Rett syndrome, Tuberous Sclerosis complex), multiple sclerosis, sterol synthesis disorders, pain (including acute and chronic pain), encephalopathy secondary to a medical condition (including hepatic encephalopathy and anti-NMDA receptor encephalitis), seizure disorder (including status epilepticus and monogenic forms of epilepsy such as Dravet's disease), stroke, traumatic brain injury, movement disorder (including Huntington's disease and Parkinson's disease), vision impairment, hearing loss, or tinnitus.

148. The method according to claim 139, wherein the disorder is a sterol synthesis disorder.

149. A compound of Formula (I-62): ##STR00666## or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 is hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl; each of R.sup.2 and R.sup.3 is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, or 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 substituted or unsubstituted membered ring; each of R.sup.4 and R.sup.5 is independently hydrogen, halo, or —OR.sup.C, wherein R.sup.C is hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl, or R.sup.4 and R.sup.5, together with the carbon atom to which they are attached form an oxo group; R.sup.6 is absent or hydrogen; and represents a single or double bond, wherein when one of is a double bond, the other is a single bond; when both of are single bonds, then R.sup.6 is hydrogen; and when one of is a double bond, R.sup.6 is absent.

150. The compound of claim 149, wherein R.sup.1 is C.sub.1-C.sub.6 alkyl.

151. The compound of claim 149, wherein R.sup.1 is substituted or unsubstituted C.sub.2-C.sub.6 alkyl.

152. The compound of claim 149, wherein R.sup.1 is substituted or unsubstituted methyl or substituted or unsubstituted ethyl.

153. The compound of claim 149, wherein R.sup.1 is substituted or unsubstituted methyl or substituted or unsubstituted ethyl.

154. The compound of claim 149, wherein R.sup.1 is trifluoromethyl.

155. The compound of claim 149, wherein R.sup.1 is —CH.sub.2OR.sup.A, wherein R.sup.A is C.sub.1-C.sub.6 alkyl.

156. The compound of claim 149, wherein R.sup.2 is hydrogen or C.sub.1-C.sub.6 alkyl.

157. The compound of claim 149, wherein R.sup.2 is hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl.

158. The compound of claim 149, wherein R.sup.2 is C.sub.1-C.sub.6 haloalkyl.

159. The compound of claim 149, wherein each of R.sup.2 and R.sup.3 is independently hydrogen or C.sub.1-C.sub.6 alkyl.

160. The compound of claim 149, wherein each of R.sup.2 and R.sup.3 is independently hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl.

161. The compound of claim 149, wherein each of R.sup.2 and R.sup.3 is independently C.sub.1-C.sub.6 alkyl or hydrogen.

162. The compound of claim 149, wherein each of R.sup.2 and R.sup.3 is independently substituted or unsubstituted C.sub.1-C.sub.6 alkyl or hydrogen.

163. The compound of claim 149, wherein R.sup.2 and R.sup.3, together with the carbon atom to which they are attached, form a 3-8 membered ring.

164. The compound of claim 149, wherein R.sup.4 is —OH or halo.

165. The compound of claim 149, wherein R.sup.4 and R.sup.5, together with the carbon atom to which they are attached form an oxo group.

166. The compound of claim 149, wherein R.sup.4 is hydrogen and R.sup.5 is halo.

167. The compound of claim 149, wherein R.sup.4 and R.sup.5 are halo.

168. The compound of claim 149, wherein R.sup.4 and R.sup.5 are hydrogen.

169. The compound of claim 149, wherein R.sup.1 is substituted or unsubstituted ethyl and R.sup.2 and R.sup.3 are substituted or unsubstituted methyl.

170. The compound of claim 149, wherein R.sup.1 is substituted or unsubstituted ethyl and R.sup.2 and R.sup.3 are substituted or unsubstituted methyl.

171. The compound of claim 149, wherein the compound of Formula (I-62) is a compound of Formula (I-A62), (I-B62), or (I-C62): ##STR00667##

172. The compound of claim 149, wherein the compound of Formula (I-62) is selected from a compound of Formula (I-C62): ##STR00668##

173. The compound of claim 149, wherein the compound of Formula (I-62) is selected from a compound of Formula (I-A62): ##STR00669##

174. The compound of claim 149, wherein R.sup.1 is substituted or unsubstituted ethyl and R.sup.2 and R.sup.3 are substituted or unsubstituted methyl.

175. The compound of claim 149, wherein R.sup.1 is substituted or unsubstituted ethyl and R.sup.2 and R.sup.3 are substituted or unsubstituted methyl.

176. The compound of claim 149, wherein the compound of Formula (I-62) is selected from a compound of Formula (I-C-i62) or (I-C-ii62): ##STR00670##

177. The compound of claim 149, wherein the compound is ##STR00671##

178. A pharmaceutical composition comprising a compound of claim 149, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

179. A method for treating or preventing a disorder described herein, comprising administering to a subject in need thereof an effective amount of a compound of claim 149, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof.

180. The method according to claim 179, wherein the disorder is a gastrointestinal (GI) disorder, constipation, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, structural disorders affecting the GI, anal disorders, hemorrhoids, internal hemorrhoids, external hemorrhoids, anal fissures, perianal abscesses, anal fistula, colon polyps, cancer, or colitis.

181. The method according to claim 179, wherein the disorder is inflammatory bowel disease.

182. The method according to claim 179, wherein the disorder is cancer, diabetes, or a sterol synthesis disorder.

183. The method according to claim 179, wherein the disorder is a metabolic disorder.

184. The method according to claim 179, wherein the disorder is an autoimmune disorder.

185. The method according to claim 179, wherein the disorder is rheumatoid arthritis, juvenile idiopathic arthritis, ankylosing spondylitis, psoriatic arthritis, Crohn's disease, ulcerative colitis, and plaque psoriasis.

186. A method for treating or preventing a CNS-related condition comprising administering to a subject in need thereof an effective amount of a compound of claim 149, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof.

187. The method according to claim 186, wherein the CNS-related condition is an adjustment disorder, anxiety disorder (including obsessive-compulsive disorder, posttraumatic stress disorder, and social phobia), cognitive disorder (including Alzheimer's disease and other forms of dementia), dissociative disorder, eating disorder, mood disorder (including depression and postpartum depression), bipolar disorder, dysthymic disorder, suicidality), schizophrenia or other psychotic disorder (including schizoaffective disorder), sleep disorder (including insomnia), substance-related disorder, personality disorder (including obsessive-compulsive personality disorder), autism spectrum disorders (including those involving mutations to the Shank group of proteins (including Shank3)), neurodevelopmental disorder (including Rett syndrome, Tuberous Sclerosis complex), multiple sclerosis, sterol synthesis disorders, pain (including acute and chronic pain), encephalopathy secondary to a medical condition (including hepatic encephalopathy and anti-NMDA receptor encephalitis), seizure disorder (including status epilepticus and monogenic forms of epilepsy such as Dravet's disease), stroke, traumatic brain injury, movement disorder (including Huntington's disease and Parkinson's disease), vision impairment, hearing loss, or tinnitus.

188. The method according to claim 179, wherein the disorder is a sterol synthesis disorder.

189. A compound of Formula (I-60): ##STR00672## or a pharmaceutically acceptable salt thereof, wherein: each of R.sup.2 and R.sup.3 is independently hydrogen, substituted or unsubstituted C.sub.1-C.sub.6 alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, or R.sup.2 and R.sup.3, together with the carbon atom to which they are attached form a substituted or unsubstituted 3-8 membered ring; each of R.sup.4 and R.sup.5 is independently hydrogen, halo, or —OR.sup.C, wherein R.sup.C is hydrogen or substituted or unsubstituted C.sub.1-C.sub.6 alkyl, or R.sup.4 and R.sup.5, together with the carbon atom to which they are attached form an oxo group; R.sup.6 is absent or hydrogen; and represents a single or double bond, wherein when one of is a double bond, the other is a single bond; when both of are single bonds, then R.sup.6 is hydrogen; and when one of the is a double bond, R.sup.6 is absent.

190. The compound of claim 189, wherein R.sup.2 is C.sub.1-C.sub.6 alkyl or hydrogen.

191. The compound of claim 189, wherein R.sup.2 is C.sub.1-C.sub.6 haloalkyl.

192. The compound of claim 189, wherein R.sup.2 is substituted or unsubstituted C.sub.1-C.sub.6 alkyl or hydrogen.

193. The compound of claim 189, wherein R.sup.2 is aryl or heteroaryl.

194. The compound of claim 189, wherein each of R.sup.2 and R.sup.3 is independently C.sub.1-C.sub.6 alkyl or hydrogen.

195. The compound of claim 189, wherein each of R.sup.2 and R.sup.3 is independently substituted or unsubstituted C.sub.1-C.sub.6 alkyl or hydrogen.

196. The compound of claim 189, wherein each of R.sup.2 and R.sup.3 is independently unsubstituted C.sub.1-C.sub.6 alkyl) or hydrogen.

197. The compound of claim 189, wherein each of R.sup.2 and R.sup.3 is independently C.sub.1-C.sub.6 haloalkyl or hydrogen.

198. The compound of claim 189, wherein each of R.sup.2 and R.sup.3 is independently aryl or heteroaryl.

199. The compound of claim 189, wherein R.sup.2 and R.sup.3, together with the carbon atom to which they are attached form a 3-membered ring.

200. The compound of claim 199, wherein R.sup.2 and R.sup.3, together with the carbon atom to which they are attached form a cyclopropane.

201. The compound of claim 189, wherein R.sup.2 and R.sup.3, together with the carbon atom to which they are attached form a 3-8 membered carbocyclic or heterocyclic ring.

202. The compound of claim 189, wherein R.sup.2 is carbocyclyl or heterocyclyl and R.sup.3 is hydrogen.

203. The compound of claim 189, wherein R.sup.2 is trifluoromethyl and R.sup.3 is hydrogen.

204. The compound of claim 189, wherein R.sup.2 is aryl or heteroaryl and R.sup.3 is hydrogen.

205. The compound of claim 189, wherein R.sup.2 and R.sup.3 are substituted or unsubstituted methyl.

206. The compound of claim 189, wherein R.sup.2 and R.sup.3 is substituted methyl.

207. The compound of claim 189, wherein R.sup.2 and R.sup.3 is unsubstituted methyl.

208. The compound of claim 189, wherein R.sup.4 is —OH or halo

209. The compound of claim 189, wherein R.sup.4 and R.sup.5, together with the carbon atom to which they are attached form an oxo group.

210. The compound of claim 189, wherein R.sup.4 is hydrogen and R.sup.5 is halo.

211. The compound of claim 189, wherein R.sup.4 and R.sup.5 are halo.

212. The compound of claim 189, wherein R.sup.4 and R.sup.5 are hydrogen.

213. The compound of claim 189, wherein the compound of Formula (I-60) is selected from a compound of Formula (I-A60), (I-B60), or (I-C60): ##STR00673##

214. The compound of claim 189, wherein the compound of Formula (I-60) is selected from a compound of Formula (I-B60): ##STR00674##

215. The compound of claim 189, wherein at least one of R.sup.2 and R.sup.3 is C.sub.1-C.sub.6 alkyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl; or R.sup.2 and R.sup.3, together with the carbon atom to which they are attached, form a 3-8 membered ring.

216. The compound of claim 189, wherein R.sup.2 is methyl and R.sup.3 is hydrogen.

217. The compound of claim 189, wherein R.sup.2 is unsubstituted methyl and R.sup.3 is hydrogen.

218. The compound of claim 189, wherein R.sup.2 and R.sup.3 are hydrogen.

219. The compound of claim 189, wherein the compound is: ##STR00675##

220. A pharmaceutical composition comprising a compound of claim 189, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

221. A method for treating or preventing a disorder described herein, comprising administering to a subject in need thereof an effective amount of a compound of claim 189, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof.

222. The method according to claim 221, wherein the disorder is a gastrointestinal (GI) disorder e.g., constipation, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD) ulcerative colitis, Crohn's disease, structural disorders affecting the GI, anal disorders, hemorrhoids, internal hemorrhoids, external hemorrhoids, anal fissures, perianal abscesses, anal fistula, colon polyps, cancer, or colitis.

223. The method according to claim 221, wherein the disorder is inflammatory bowel disease.

224. The method according to claim 221, wherein the disorder is cancer, diabetes, or a sterol synthesis disorder.

225. The method according to claim 221, wherein the disorder is a metabolic disorder.

226. The method according to claim 221, wherein the disorder is an autoimmune disorder.

227. The method according to claim 221, wherein the disorder is rheumatoid arthritis, juvenile idiopathic arthritis, ankylosing spondylitis, psoriatic arthritis, Crohn's disease, ulcerative colitis, and plaque psoriasis.

228. A method for treating or preventing a CNS-related condition comprising administering to a subject in need thereof an effective amount of a compound of claim 189, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof.

229. The method according to claim 228, wherein the CNS-related condition is an adjustment disorder, anxiety disorder (including obsessive-compulsive disorder, posttraumatic stress disorder, and social phobia), cognitive disorder (including Alzheimer's disease and other forms of dementia), dissociative disorder, eating disorder, mood disorder (including depression and postpartum depression), bipolar disorder, dysthymic disorder, suicidality), schizophrenia or other psychotic disorder (including schizoaffective disorder), sleep disorder (including insomnia), substance-related disorder, personality disorder (including obsessive-compulsive personality disorder), autism spectrum disorders (including those involving mutations to the Shank group of proteins (including Shank3)), neurodevelopmental disorder (including Rett syndrome, Tuberous Sclerosis complex), multiple sclerosis, sterol synthesis disorders, pain (including acute and chronic pain), encephalopathy secondary to a medical condition (including hepatic encephalopathy and anti-NMDA receptor encephalitis), seizure disorder (including status epilepticus and monogenic forms of epilepsy such as Dravet's disease), stroke, traumatic brain injury, movement disorder (including Huntington's disease and Parkinson's disease), vision impairment, hearing loss, or tinnitus.

230. The method according to claim 221, wherein the disorder is a sterol synthesis disorder.

Description

EXAMPLES

[0329] In order that the invention described herein may be more fully understood, the following examples are set forth. The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions, and methods provided herein and are not to be construed in any way as limiting their scope.

[0330] Unless otherwise indicated, 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.”

[0331] The absolute configuration of an asymmetric center can be determined using methods known to one skilled in the art. In some embodiments, the absolute configuration of an asymmetric center in a compound can be elucidated from the X-ray single-crystal structure of the compound. In some embodiments, the absolute configuration of an asymmetric center elucidated by the X-ray crystal structure of a compound can be used to infer the absolute configuration of a corresponding asymmetric center in another compound obtained from the same or similar synthetic methodologies. In some embodiments, the absolute configuration of an asymmetric center elucidated by the X-ray crystal structure of a compound can be used to infer the absolute configuration of a corresponding asymmetric center in another compound coupled with a spectroscopic technique, e.g., NMR spectroscopy, e.g., .sup.1H NMR spectroscopy or .sup.19F NMR spectroscopy.

Materials and Methods

[0332] The compounds provided herein can be prepared from readily available starting materials using the following general methods and procedures. 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.

[0333] 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.

[0334] 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 following schemes are presented with details as to the preparation of representative oxysterols that have been listed herein. 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.

[0335] .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 integratations) 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.

[0336] 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.

[0337] Exemplary general method for SFC: Column: CHIRALPAK® AD CSP (250 mm 10*30 mm, 10 μm), Gradient: 45% B, A=NH.sub.3H.sub.2O, B=MeOH, flow rate: 60 mL/min. For example, AD_3_EtOH_DEA_5_40_25ML would indicate: “Column: Chiralpak AD-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp: 35° C.”.

Example 1: NMDA Potentiation

NMDA Potentiation

Whole-Cell Patch Clamp of Mammalian Cells (Ionworks Barracuda (TWB))

[0338] The whole-cell patch-clamp technique was used to investigate the effects of compounds on_GlunN1/GluN2A glutamate receptors expressed in mammalian cells.

[0339] HEK293 cells were transformed with adenovirus 5 DNA and transfected with cDNA encoding the human GRIN1/GRIN2A genes. Stable transfectants were selected using G418 and Zeocin-resistance genes incorporated into the expression plasmid and selection pressure maintained with G418 and Zeocin in the medium. Cells were cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture (D-MEM/F-12) supplemented with 10% fetal bovine serum, 100 μg/ml penicillin G sodium, 100 μg/ml streptomycin sulphate, 100 μg/ml Zeocin, 5 μg/ml blasticidin and 500 μg/ml G418.

[0340] Test article effects were evaluated in 8-point concentration-response format (4 replicate wells/concentration). All test and control solutions contained 0.3% DMSO and 0.01% Kolliphor® EL (C5135, Sigma). The test article formulations were loaded in a 384-well compound plate using an automated liquid handling system (SciClone ALH3000, Caliper LifeScienses). The measurements were performed using Ion Works Barracuda platform following this procedure:

Electrophysiological Procedures:

[0341] Intracellular solution (mM): 50 mM CsCl, 90 mM CsF, 2 mM MgCl.sub.2, 5 mM EGTA, 10 mM HEPES. Adjust to pH 7.2 with CsOH.

[0342] Extracellular solution, HB-PS (composition in mM): NaCl, 137; KCl, 1.0; CaCl.sub.2, 5; HEPES, 10; Glucose, 10; pH adjusted to 7.4 with NaOH (refrigerated until use).

[0343] Holding potential: −70 mV, potential during agonist/PAM application: −40 mV.

Recording Procedure:

[0344] Extracellular buffer is loaded into the PPC plate wells (11 μL per well). Cell suspension will be pipetted into the wells (9 μL per well) of the PPC planar electrode.

[0345] Whole-cell recording configuration is established via patch perforation with membrane currents recorded by on-board patch clamp amplifiers.

[0346] Two recordings (scans) are performed. First, during pre-application of test article alone (duration of pre-application—5 min) and second, during test articles and agonist (EC.sub.20 L-glutamate and 30 μM glycine) co-application to detect positive modulatory effects of the test article.

[0347] Test Article Administration: The first pre-application consists of the addition of μL of 2× concentrated test article solution and, second, of 20 μL of IX concentrated test article and agonist at 10 μL/s (2 second total application time).

Example 2: NAM and PAM

Whole-Cell Patch Clamp of Mammalian Cells (Ionworks Barracuda (TWB))

[0348] The whole-cell patch-clamp technique was used to investigate the effects of positive allosteric modulating activity of test compounds on_GlunN1/GluN2A and GluN2B glutamate receptors expressed in mammalian cells.

[0349] HEK293 cells were transformed with adenovirus 5 DNA and transfected with cDNA encoding the human GRIN1/GRIN2A genes. Stable transfectants were selected using G418 and Zeocin-resistance genes incorporated into the expression plasmid and selection pressure maintained with G418 and Zeocin in the medium. Cells were cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture (D-MEM/F-12) supplemented with 10% fetal bovine serum, 100 μg/ml penicillin G sodium, 100 μg/ml streptomycin sulphate, 100 μg/ml Zeocin, 5 μg/ml blasticidin and 500 μg/ml G418.

[0350] Test article effects were evaluated in 8-point concentration-response format (4 replicate wells/concentration). All test and control solutions contained 0.3% DMSO and 0.01% Kolliphor® EL (C5135, Sigma). The test article formulations were loaded in a 384-well compound plate using an automated liquid handling system (SciClone ALH3000, Caliper LifeScienses). The measurements were performed using Ion Works Barracuda platform following this procedure:

Electrophysiological Procedures:

[0351] a) Intracellular solution (mM): 50 mM CsCl, 90 mM CsF, 2 mM MgCl.sub.2, 5 mM EGTA, 10 mM HEPES. Adjust to pH 7.2 with CsOH. [0352] b) Extracellular solution, HB-PS (composition in mM): NaCl, 137; KCl, 1.0; CaCl.sub.2, 5; HEPES, 10; Glucose, 10; pH adjusted to 7.4 with NaOH (refrigerated until use). [0353] c) Holding potential: −70 mV, potential during agonist/PAM application: −40 mV.

Recording Procedure:

[0354] a) Extracellular buffer will be loaded into the PPC plate wells (11 μL per well). Cell suspension will be pipetted into the wells (9 μL per well) of the PPC planar electrode. [0355] b) Whole-cell recording configuration will be established via patch perforation with membrane currents recorded by on-board patch clamp amplifiers. [0356] c) Two recordings (scans) will be performed. First, during pre-application of test article alone (duration of pre-application—5 min) and second, during test articles and agonist (EC.sub.20 L-glutamate and 30 μM glycine) co-application to detect positive modulatory effects of the test article.

[0357] Test Article Administration: The first pre-application will consist of the addition of 20 μL of 2× concentrated test article solution and, second, of 20 μL of IX concentrated test article and agonist at 10 μL/s (2 second total application time).

Potentiating Effect of Positive Allosteric Modulators (PAM) on the Channel

[0358] Potentiating effect of positive allosteric modulators (PAM) on the channel will be calculated as

% activation=(I.sub.PAM/I.sub.EC10-30)×100%−100%

where I.sub.PAM will be the L-glutamate EC.sub.10-30—elicited current in presence of various concentrations of test articles and I.sub.EC20 will be the mean current elicited with L-glutamate EC.sub.20. PAM concentration-response data will be fitted to an equation of the form:

% Activation=% L-glutamate EC.sub.20+{(% MAX−% L-glutamate EC.sub.20)/[1+([Test]/EC.sub.50).sup.N]},

where [Test] will be the concentration of PAM (test article), EC.sub.50 will be the concentration of PAM producing half-maximal activation, N will be the Hill coefficient, % L-glutamate EC.sub.20 will be the percentage of the current Elicited with L-glutamate EC.sub.20, % MAX is the percentage of the current activated with the highest dose of PAM co-admitted with L-glutamate EC.sub.20 and % Activation will be the percentage of the current elicited with L-glutamate EC.sub.10-30 at each PAM concentration.

[0359] The maximal amplitude of the evoked currents are measured and defined as Peak Current Amplitude (PCA).

Automated Patch-Clamp System (QPatch HTX):

[0360] In this study, HEK 293 cells stably transfected with glutamate-activated channels of the GRIN1/2A subtype will be used together with submaximal NMDA concentrations (300 μM NMDA, co-application with 8 μM Glycine) to investigate the negative allosteric modulation of the test compounds.

Cell Culture

[0361] In general, cells will be passaged at a confluence of about 80% to-90%. For electrophysiological measurements cells will be harvested at a confluence of about 80% to 90% from sterile culture flasks containing culture complete medium. Cells will be transferred as suspension in PBS to the QPatch 16X or QPatch HTX system to the centrifuge/washer directly.

[0362] Standard Laboratory Conditions: Cells will be incubated at 37° C. in a humidified atmosphere with 5% CO.sub.2 (rel. humidity about 95%).

[0363] Culture Media:

[0364] The cells will be continuously maintained in and passaged in sterile culture flasks containing a 1:1 mixture of Dulbecco's modified eagle medium and nutrient mixture F-12 (D-MEM/F-12 1×, liquid, with L-Glutamine) supplemented with 10% fetal bovine serum, 1% Penicillin/Streptomycin solution, and 50 μM AP-5 blocker.

[0365] Antibiotics:

[0366] The complete medium as indicated above is supplemented with 100 μg/mL hygromycin, 15 μg/mL blasticidin and 1 μg/mL puromycin.

[0367] Induction of Expression:

[0368] 2.5 μg/mL tetracycline is added 24 h before start of experiments.

Dose Formulation

[0369] Dose levels are in terms of test compounds, as supplied. Vehicle will be added to achieve a stock concentration of 10 mM (storage at −10° C. to −30° C.). A further stock solutions of 1.0 mM will be prepared in DMSO. Details of stock solution usage (thawing, dose formulations) will be documented in the raw data. The time period of stock solution usage will be detailed in the report.

Test Compound Concentrations

[0370] Dose levels are in terms of test compounds, as supplied. Vehicle will be added to achieve a stock concentration of 10 mM (storage at −10° C. to −30° C.). A further stock solutions of 1.0 mM will be prepared in DMSO. Details of stock solution usage (thawing, dose formulations) will be documented in the raw data. The time period of stock solution usage will be detailed in the report.

[0371] One test concentration of 1.0 μM will be tested.

[0372] All test solutions will be prepared by diluting the stock solutions with either Mg-free bath solution only or Mg-free bath solution containing NMDA (300 μM) and glycine (8.0 μM) shortly prior to the electrophysiological experiments and kept at room temperature (19° C. to 30° C.) when in use. 0.1% DMSO will be used as vehicle.

[0373] Frequency of Preparation:

[0374] For each test concentration, fresh solutions of test compounds will be prepared every day.

[0375] Stability of Dose Formulation:

[0376] All preparation times will be documented in the raw data. Any observations regarding instability of test compounds will be mentioned in the raw data.

[0377] Storage of Dose Formulation:

[0378] On the day of experimentation dose formulations will be maintained at room temperature (19° C. to 30° C.) when in use.

Bath Solutions

[0379] For preparing the experiments and for formation of the giga-ohm-seal, the following standard bath solution will be used:

[0380] Sodium Chloride: 137 mM; Potassium Chloride: 4 mM; Calcium Chloride: 1.8 mM; Magnesium Chloride: 1 mM; HEPES: 10 mM; D-Glucose: 10 mM; Cremophor: 0.02%; pH (NaOH): 7.4

[0381] The 1× bath solution will be prepared by diluting 10× bath solution without Glucose and 100× Glucose solution with water at least every 7 days. Both stock solutions have been prepared prior to the experimental start of the present study and stored at 1° C. to 9° C. (10× bath solution) or −10° C. to −30° (100× Glucose solution). The batch number(s) of the bath solution(s) used in the experiments will be documented in the raw data. When in use, the 1× bath solution will be kept at room temperature (19° C. to 30° C.). When not in use, the 1× bath solution will be stored at 1° C. to 9° C.

[0382] After the giga-seal was formed the following Mg-free bath solution will be used:

[0383] Sodium Chloride: 137 mM; Potassium Chloride: 4 mM; Calcium Chloride; 2.8 mM; HEPES: 10 mM; D-Glucose: 10 mM; Cremophor: 0.02%; pH (NaOH): 7.4 This Mg-free bath solution will be prepared as a 1× solution and stored at 1° C. to 9° C. It will be prepared freshly at least every 10 days.

Intracellular Solution

[0384] The 1× intracellular solution will be thawed every day out of a frozen 1× intracellular solution, which has been prepared prior to the experimental start of the present study, aliquoted and stored at −10° C. to −30° C. When in use, the 1× intracellular solution will kept at room temperature (19° C. to 30° C.). Remaining 1× intracellular solution will be stored in the fridge (1° C. to 9° C.). The 1× intracellular solution will include the components outlined below:

[0385] Potassium Chloride: 130 mM; Magnesium Chloride: 1 mM; Mg-ATP: 5 mM; HEPES: 10 mM; EGTA: 5 mM; pH (KOH): 7.2

Cell Treatment

[0386] For this study, cells will continuously be perfused with NMDA/Glycine, Test Compound or Test Compound/NMDA/Glycin.

[0387] In every case, at least 30-second prewash steps with a test compound will be performed in between applications. For details see Table A below.

[0388] Each experiment type will be analyzed in at least n=3 isolated cells. The NMDA and Glycine stock solutions will be prepared prior to the experimental start of the present study, stored frozen (−10° C. to −30° C.) until the day of experimentation. Shortly prior to the electrophysiological experiments, frozen stock solutions will be thawed and diluted.

[0389] Control: The effect of vehicle (0.1% DMSO) and D-(−)-2-Amino-5-phosphonopentanoic acid (AP-5) (100 μM) will be measured at three cells every second week, in order to assure successful expression of NMDA receptors.

[0390] The 50 mM stock solution of AP-5 has been prepared prior to the experimental start of the present study, aliquoted and stored frozen (−10° C. to −30° C.) until the day of experimentation. Shortly prior to the electrophysiological experiments the frozen stock solution will be thawed and then diluted in Mg-free bath solution containing NMDA (300 μM) and glycine (8.0 μM), to give a final perfusion concentration of 100 μM.

Experimental Procedure

[0391] Cells are transferred as suspension in serum-free medium to the QPatch HTX system and kept in the cell storage tank/stirrer during experiments. All solutions applied to cells including the intracellular solution will be maintained at room temperature (19° C. to 30° C.).

[0392] During the sealing process standard bath solution described above will be used. All solutions applied to cells including the pipette solution will be maintained at room temperature (19° C. to 30° C.). After formation of a Gigaohm seal between the patch electrodes and transfected individual HEK293 cells only Mg-free bath solution will be perfused and the cell membrane will be ruptured to assure electrical access to the cell interior (whole-cell patch-configuration). Inward currents will be measured upon application of 300 μM NMDA (and 8.0 μM Glycine) to patch-clamped cells for 5 sec. During the entire experiment the cells will be voltage-clamped at a holding potential of −80 mV.

[0393] For the analysis of test compounds, NMDA receptors will be stimulated by 300 μM NMDA and 8.0 μM Glycine and test compound combinations described below. Thirty-second prewash steps with a test compound will be performed in between applications.

TABLE-US-00001 TABLE A Application Protocol; use dependence of test compounds Appl. # Duration (s) Application 1  4 NMDA/Glycine 2 30 Bath 3  4 NMDA/Glycine 2 repetitions 4 30 1 μM Test Compound 5  4 1 μM Test Compound + NMDA/Glycine 6 repetitions 6 30 Bath 7  4 NMDA/Glycine 2 repetitions

TABLE-US-00002 TABLE B Application Protocol; control experiments Appl. # Duration (s) Application 1  4 NMDA/Glycine 2 30 Bath 3  4 NMDA/Glycine 2 repetitions 4 30 Bath 5  4 NMDA/Glycine 6 repetitions 6 30 Bath 7  4 NMDA/Glycine + 100 μM AP-5 2 repetitions

Example 3. Synthesis of Compound 1

[0394] ##STR00078##

Step 1

[0395] To a mixture of MePPh.sub.3Br (1.28 kg, 3.6 mol) in THF (4.5 L) was added t-BuOK (404 g, 3.6 mol) at 15° C. under N.sub.2. The resulting mixture was stirred at 50° C. for 30 mins. Pregnenolone (950 g, 2.9 mol) was added in portions below 65° C. The reaction mixture was stirred at 50° C. for 1 hour. The combined mixture was quenched with saturated NH.sub.4Cl aqueous (1 L) at 15° C. THF layer was separated. The aqueous was extracted with EtOAc (2×2 L). The combined organic phase was concentrated under vacuum to give a solid. The solid was further purified by trituration with MeOH/H.sub.2O (1:1, 15 L) at reflux to give A-1 (940 g, 99%) as a solid.

[0396] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.32 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H), 3.58-3.46 (m, 1H), 2.36-2.16 (m, 2H), 2.08-1.94 (m, 2H), 1.92-1.62 (m, 9H), 1.61-1.39 (m, 6H), 1.29-1.03 (m, 4H), 1.01 (s, 3H), 0.99-0.91 (m, 1H), 0.59 (s, 3H).

Step 2

[0397] To a solution of A-1 (800 g, 2.54 mol) in DCM (8 L) was added DMP (2.14 kg, 5.08 mol) in portions at 35° C. The reaction mixture was stirred at 35° C. for 20 mins. The reaction mixture was filtered. The filtered cake was washed with DCM (3 xl L). The combined organic phase was washed with saturated Na.sub.2S.sub.2O.sub.3/saturated NaHCO.sub.3 aqueous (3:1, 2×1.5 L), brine (1.5 L), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give A-2 (794 g, crude) as a solid, which was used for next step directly.

Step 3

[0398] To a solution of TBAF (3.04 mL, 1 M in THF, 3.04 mmol, Aldrich) in THF (100 mL) was added TMSCF.sub.3 (25.8 g, 182 mmol) followed by a solution of A-2 (19 g, 60.8 mmol) in THF (100 mL) dropwise at 0° C. The mixture was stirred at 0° C. for 30 mins. To the mixture was added TBAF (200 mL, 1 M in THF, 200 mmol) at 0° C. The mixture was stirred at 0° C. for another 30 mins. To the mixture was added saturated aqueous NH.sub.4Cl (100 mL) and the mixture was concentrated in vacuum. To the residue was added PE/EtOAc (400 mL, 1:1), the organic layer was separated, which was combined with other two batches (2×10 g of A-2). The combined organic layer was washed with water (300 mL), brine (300 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give an oil. The residue was dissolved in DCM (150 mL) and diluted with PE (750 mL). The solution was poured into a silica gel column (500 g, 100˜200 mesh) and eluted with PE:DCM:EtOAc=5:1:0.05 to 5:1:0.1 to give A-4 (12 g, 17% yield) as an oil and impure A-3. The impure A-3 was re-crystallized from MeCN (250 mL) to give purified A-3 (6.5 g) as a solid. A-3 recovered from the MeCN filtrate was subjected to silica gel chromatography (PE:DCM:EtOAc=50:1:1 to 20:1:1) to give a crude product which was re-crystallized from MeCN (20 mL) to give purified A-3 (1 g, 16% total yield) as a solid.

[0399] Note: A-3 and A-4 were identified from .sup.3J.sub.H,CF, (FDCS). (J. Org. Chem. 2015, 80, 1754.).

[0400] A-3:

[0401] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.43-5.33 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H); 2.49 (s, 2H); 2.11-1.97 (m, 4H), 1.95-1.32 (m, 14H), 1.30-0.98 (m, 7H), 0.59 (s, 3H).

[0402] A-4:

[0403] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.54-5.41 (m, 1H), 4.86 (s, 1H), 4.72 (s, 1H); 2.78-2.65 (m, 1H); 2.18-1.97 (m, 3H), 1.95-1.35 (m, 16H), 1.32-0.98 (m, 7H), 0.59 (s, 3H).

Step 4

[0404] To a solution of A-3 (8 g, 20.9 mmol) in THF (80 mL) was added 9-BBN dimer (5.85 g, 24 mmol). The mixture was stirred at 40° C. for 1 h. The mixture was cooled to 0° C. To the mixture was added EtOH (12 mL), NaOH (41.8 mL, 5 M, aq.) and H.sub.2O.sub.2 (20.9 mL, 10 M, aq.) dropwise. The mixture was stirred at 50° C. for 1 h and then cooled. To the mixture was added Na.sub.2SO.sub.3 (100 mL, 25%, aq.). The mixture was extracted with EtOAc (300 mL). The organic layer was separated, purified by silica gel column (PE:EtOAc=10:1 to 5:1) to give A-5 (7.1 g, 85%) as a solid.

[0405] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.32 (m, 1H), 3.64 (dd, J=3.2, 10.4 Hz, 1H), 3.37 (dd, J=6.8, 10.4 Hz, 1H), 2.49 (s, 2H), 2.32-1.92 (m, 4H), 1.92-1.70 (m, 4H), 1.70-1.29 (m, 8H), 1.29-0.91 (m, 11H), 0.71 (s, 3H).

Step 5

[0406] To a solution of A-5 (7.1 g, 17.7 mmol) in DCM (30 mL) and pyridine (21 mL) was added TsCl (6.74 g, 35.4 mmol). The mixture was stirred at 15° C. for 2 hrs. To the mixture was added water (5 mL) and the mixture was stirred at 15° C. for 2 hrs. The mixture was concentrated in vacuum. To the residue was added water (100 mL) and EtOAc (200 mL). The organic layer was separated, washed with HCl (100 mL, 0.1 M), water (100 mL) and brine (100 mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give A-6 (9.8 g, 100%) as a solid.

[0407] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.78 (d, J=8.0 Hz, 2H), 7.34 (d, J=8.0 Hz, 2H), 5.48-5.29 (m, 1H), 3.97 (dd, J=2.4, 9.2 Hz, 1H), 3.77 (dd, 7=6.4, 9.2 Hz, 1H), 2.48 (s, 2H), 2.45 (s, 3H), 2.10-1.88 (m, 5H), 1.82-1.35 (m, 9H), 1.30-0.82 (m, 12H), 0.64 (s, 3H).

Step 6

[0408] To a solution of A-6 (1.05 g, 1.89 mmol) in DMF (5 mL) was added KI (1.25 g, 7.56 mmol). The mixture was stirred at 50° C. for 1 h. To the mixture was added PhSO.sub.2Na (0.93 g, 5.67 mmol). The mixture was stirred at 50° C. for 2 hrs. To the mixture was added water (10 mL) and DCM (30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum and triturated form PE/DCM (10 mL, 5:1) to give A-7 (600 mg, 61%) as a solid.

[0409] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.98-7.87 (m, 2H), 7.70-7.52 (m, 3H), 5.39-5.31 (m, 1H), 3.14 (d, J=14.0 Hz, 1H), 2.85 (dd, J=9.6, 14.0 Hz, 1H), 2.48 (s, 2H), 2.20-1.88 (m, 5H), 1.88-1.68 (m, 4H), 1.60-1.33 (m, 5H), 1.30-0.82 (m, 12H), 0.66 (s, 3H).

Step 7

[0410] To a solution of i-Pr.sub.2NH (576 mg, 5.70 mmol) in THF (10 mL) was added n-BuLi (1.9 mL, 2.5 M in hexane, 4.75 mmol) at −70° C. The mixture was warmed to 0° C. A solution of A-7 (1 g, 1.9 mmol) in THF (8 mL) was added at −70° C. The mixture was stirred at −70° C. for 1 h. To the mixture was added a solution of 2-isopropyloxirane (245 mg, 2.85 mmol) in THF (2 mL) at −70° C. The mixture was stirred at −70° C. for 1 h, warmed to 10° C. and stirred at 10° C. for 16 hrs. To the mixture was added NH.sub.4Cl (5 mL, sat. aq.). The mixture was extracted with EtOAc (50 mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give A-8 (1.2 g crude) as an oil.

Step 8

[0411] To a solution of A-8 (1.2 g, 1.96 mmol) in MeOH (60 mL) was added NiBr.sub.2 (5 mg, 0.023 mmol) and Mg powder (3.79 g, 156 mmol) was added in portions within 30 mins at 60° C. The mixture was stirred at 60° C. for 10 mins. The mixture was poured into HCl (160 mL, 2 M) and extracted with PE/EtOAc (2×200 mL, 1:1). The combined organic layer was washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum and purified by silica gel column (100˜200 mesh, PE:EtOAc=50:1 to 10:1) twice to give a crude product, which was purified by silica gel column (200˜300 mesh, PE:DCM:acetone=1:1:0.01) twice, re-crystallized from MeCN/water (3:1, 5 mL) to give Compound 1 (50 mg, 5%) as a solid.

[0412] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.41-5.32 (m, 1H), 3.39-3.28 (m, 1H), 2.49 (s, 2H), 2.10-1.92 (m, 4H), 1.90-1.60 (m, 5H), 1.55-1.33 (m, 8H), 1.31-1.10 (m, 6H), 1.09-0.90 (m, 15H), 0.68 (s, 3H).

[0413] LCMS Rt=1.278 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.28H.sub.44F.sub.3O [M+H−H.sub.2O].sup.+ 453, found 453.

Example 4. Syntheses of Compounds 1-A and 1-B

[0414] ##STR00079##

Step 1

[0415] To a solution of Compound 1 (100 mg, 0.212 mmol) in pyridine (3 mL) was added benzoyl chloride (59.7 mg, 0.425 mmol) at 25° C. The reaction was stirred at 25° C. for 16 hrs. The reaction was quenched by water (10 mL) and extracted with EtOAc (2×10 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give crude product. The crude product was purified by a silica gel column (PE/EtOAc=10/1) to give desired product A-9 (150 mg, crude) as a solid.

[0416] LCMS Rt=1.544 min in 2 min chromatography, 30-90 AB, MS ESI calcd. For C.sub.35H.sub.49F.sub.3O.sub.3 [M+Na]+ 597, found 597.

Step 2

[0417] A-9 (580 mg, 1.00 mmol) was purified by SFC separation (column: AD (250 mm*30 mm, 5 um), gradient: 45% B (A=NH.sub.3H.sub.2O, B=MeOH), flow rate: 60 mL/min) to give A-10-A (200 mg, 34%, 95.5% d.e. by SFC (Column: Chiralpak AD-3 100×4.6 mm I.D., 3 um, Mobile phase: A: CO.sub.2 B: iso-propanol (0.05% DEA).

[0418] Gradient: from 5% to 40% of B in 4.5 min and hold 40%, for 2.5 min, then 5% of B for 1 min.

[0419] Flow rate: 2.8 mL/min Column temperature: 40° C.)) as a solid and A-10-B (215 mg, 37%, 99.5% d.e. by SFC (Column: Chiralpak AD-3 100×4.6 mm I.D., 3 um, Mobile phase: A: CO.sub.2 B: iso-propanol (0.05% DEA).

[0420] Gradient: from 5% to 40% of B in 4.5 min and hold 40%, for 2.5 min, then 5% of B for 1 min. Flow rate: 2.8 mL/min Column temperature: 40° C.)) as a solid.

[0421] A-10-A:

[0422] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.07-8.02 (m, 2H), 7.58-7.52 (m, 1H), 7.48-7.41 (m, 2H), 5.37-5.35 (m, 1H), 4.99-4.94 (m, 1H), 2.48-2.46 (m, 2H), 2.04-1.89 (m, 4H), 1.82-1.65 (m, 5H), 1.51-1.35 (m, 7H), 1.27-1.08 (m, 4H), 1.05 (s, 4H), 1.02-0.92 (m, 13H), 0.64 (s, 3H).

[0423] A-10-B:

[0424] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.07-8.02 (m, 2H), 7.59-7.52 (m, 1H), 7.49-7.40 (m, 2H), 5.37-5.35 (m, 1H), 5.01-4.92 (m, 1H), 2.48-2.46 (m, 2H), 2.03-1.90 (m, 5H), 1.83-1.66 (m, 3H), 1.83-1.66 (m, 1H), 1.51-1.37 (m, 8H), 1.23-1.11 (m, 3H), 1.05-1.00 (m, 5H), 0.99-0.90 (m, 12H), 0.66 (s, 3H).

Step 2a

[0425] To a solution of A-10-A (215 mg, 0.374 mmol) in THF (2 mL) and MeOH (2 mL) was added NaOH (400 mg, 10 mmol) and H.sub.2O (2 mL) at 25° C. The solution was stirred at 50° C. for 48 hrs. The reaction solution was extracted with EtOAc (2×10 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give crude product which was triturated with MeCN (2×5 mL) to give desired product Compound 1-A (148 mg, 84%) as a solid.

[0426] Compound 1-A:

[0427] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.38-5.36 (m, 1H), 3.33-3.31 (m, 1H), 2.49-2.48 (m, 2H), 2.08-1.92 (m, 4H), 1.89-1.61 (m, 5H), 1.52-1.37 (m, 5H), 1.32-1.09 (m, 7H), 1.06-0.96 (m, 7H), 0.96-0.87 (m, 10H), 0.68 (s, 3H). LCMS Rt=1.497 min in 2 min chromatography, 30-90 AB, MS ESI calcd. For C.sub.28H.sub.44F.sub.3O [M+H−H.sub.2O]+ 453, found 453.

Step 2b

[0428] To a solution of A-10-B (200 mg, 0.348 mmol) in THF (2 mL) and MeOH (2 mL) was added NaOH (400 mg, 10 mmol) and H.sub.2O (2 mL) at 25° C. The solution was stirred at 50° C. for 48 hrs. The reaction solution was extracted with EtOAc (2×10 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give crude product, which was triturated with MeCN (2×5 mL) to give desired product Compound 1-B (139 mg, 85%) as a solid.

[0429] Compound 1-B:

[0430] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.38-5.36 (m, 1H), 3.33-3.31 (m, 1H), 2.49-2.48 (m, 2H), 2.12-1.92 (m, 5H), 1.89-1.40 (m, 12H), 1.29-1.11 (m, 5H), 1.09-0.98 (m, 6H), 0.95-0.89 (m, 10H), 0.69 (s, 3H)

[0431] LCMS Rt=1.500 min in 2 min chromatography, 30-90 AB, MS ESI calcd. For C.sub.28H.sub.44F.sub.3O [M+H−H.sub.2O].sup.+ 453, found 453.

Synthesis of Compound 1-A—Absolute Stereochemistry

[0432] ##STR00080## ##STR00081##

[0433] The experimental procedures of intermediate ST-200-CF3_4A or A-7 can be found in Example 3.

Synthesis of ST-200-096-004_1

[0434] ##STR00082##

[0435] To a solution of ST-200-096-004_1 (450 mg, 0.736 mmol) in methanol (30 mL) was added Mg powder (883 mg, 36.8 mmol) under N.sub.2 at 65° C. The reaction mixture was quenched with HCl (50 mL) dropwise until the solution became clear. The reaction solution was extracted with EtOAc (3×30 mL). The combined organic layer was washed with sat. NaHCO.sub.3 (50 mL), brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-12% of EtOAc in PE) to give ST-200-096-004_2 (150 mg, 43%) as a solid.

[0436] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.34 (m, 1H), 3.37-3.25 (m, 1H), 2.55-2.40 (m, 2H), 2.09-1.91 (m, 4H), 1.90-1.70 (m, 3H), 1.69-1.56 (m, 4H), 1.54-1.35 (m, 6H), 1.34-0.97 (m, 12H), 0.96-0.86 (m, 9H), 0.68 (s, 3H).

Synthesis of ST-200-096-004_3

[0437] ##STR00083##

[0438] To a solution of ST-200-096-004_2 (150 mg, 0.318 mmol) in pyridine (3 mL) was added BzCl (134 mg, 0.954 mmol) at 0° C. and the reaction was stirred at 25° C. for 2 h. The reaction mixture was diluted with water (50 mL), extracted with EtOAc (2×40 mL). The organic layer was washed with brine (5×50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The crude was purified by silica gel column (PE/EtOAc=10/1 to 4/1) to give ST-200-096-004_3 (120 mg, 66%) as a solid.

[0439] The ST-200-096-004_3 (120 mg, 0.208 mmol) was separated by SFC (column: AD (250 mm*30 mm, 5 um)), gradient: 25-25% B (0.1% NH.sub.3H.sub.2O IPA)) to give ST-200-096-004_4 (100 mg, 84%) as a solid.

[0440] .sup.1HNMR (400 MHz, CDCl3) δ 8.05 (d, J=8 Hz, 2H), 7.55 (t, J=8 Hz, 1H), 7.44 (t, J=8 Hz, 2H), 5.38-5.34 (m, 1H), 4.98-4.91 (m, 1H), 2.48 (s, 2H), 2.09-1.89 (m, 4H), 1.86-1.67 (m, 4H), 1.53-1.34 (m, 10H), 1.17-1.00 (m, 7H), 0.99-0.91 (m, 12H), 0.64 (s, 3H).

[0441] SFC Rt=3.473 min in 10 min chromatography, AD_IPA (DEA)_5_40_2, 8ML_8MIN, 100% de.

Synthesis of Compound 1-A

[0442] ##STR00084##

[0443] To a solution of ST-200-096-004_4 (100 mg, 0.173 mmol) in THF (2 mL) and MeOH (1 mL) and water(l mL) was added KOH (48.5 mg, 0.865 mmol). The mixture was stirred at 60° C. for 16 hrs. The mixture was poured into water (20 mL) and extracted with EtOAc (2×40 mL). The combined organic layer was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (PE/EtOAc=5/l to 3/1) to give Compound 1-A (48 mg, 59%) as a solid.

[0444] .sup.1HNMR (400 MHz, CDCl3) δ 5.40-5.35 (m, 1H), 3.35-3.28 (m, 1H), 2.49 (m, 2H), 2.09-1.93 (m, 4H), 1.89-1.59 (m, 6H), 1.54-1.22 (m, 10H), 1.20-0.97 (m, 9H), 0.95-0.89 (m, 9H), 0.68 (s, 3H).

[0445] LCMS Rt=1.265 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.28H.sub.44F.sub.3O [M−H.sub.2O+H]=453, found 453.

Example 5. Syntheses of Compounds 2, 2-A, and 2-B

[0446] ##STR00085## ##STR00086## ##STR00087##

Step 1

[0447] To a solution of pregnenolone (50 g, 157 mmol) in THF (750 mL) and MeOH (500 mL) was added Pd/C (20 g, 10%, dry). The mixture was stirred under H.sub.2 (25 psi) at 25° C. for 72 hrs. The mixture was filtered. The filtrate was concentrated in vacuum to give B-1 (47 g, 94%) as a solid.

[0448] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.69-3.51 (m, 1H), 2.51 (t, J=8.8 Hz, 1H), 2.21-2.12 (m, 1H), 2.11 (s, 3H), 2.05-1.98 (m, 1H), 1.88-1.77 (m, 1H), 1.77-1.53 (m, 6H), 1.48-1.08 (m, 11H), 1.05-0.85 (m, 2H), 0.80 (s, 3H), 0.73-0.63 (m, 1H), 0.60 (s, 3H).

Step 2

[0449] To a suspension of MePPh.sub.3Br (78.5 g, 220 mmol) in THF (250 mL) was added t-BuOK (24.6 g, 220 mmol). The mixture was stirred at 50° C. for 1 h. To the mixture was added B-1 (47 g, 147 mmol). The mixture was stirred at 50° C. for 1 h. To the mixture was added water (100 mL) and EA (500 mL). The organic layer was separated, concentrated in vacuum to give a crude product, which was triturated from MeOH/water (1000 mL, 1:1) at 50° C. The mixture was filtered after cooled and the solid was washed with MeOH/water (2×500 mL, 1:1), dried in vacuum to give B-2 (45 g, 97%) as a solid.

[0450] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.84 (s, 1H), 4.70 (s, 1H), 3.69-3.51 (m, 1H), 2.08-1.98 (m, 1H), 1.88-1.62 (m, 10H), 1.61-1.50 (m, 2H), 1.48-0.85 (m, 13H), 0.81 (s, 3H), 0.70-0.60 (m, 1H), 0.56 (s, 3H).

Step 3

[0451] To a solution of B-2 (45 g, 142 mmol) in DCM (500 mL) was added silica gel (90 g) and PCC (45.7 g, 213 mmol). The mixture was stirred at 20° C. for 3 hrs. To the mixture was added PE (500 mL). The mixture was filtered though a pad of silica gel and the solid was washed with PE/DCM (1:1, 2 L). The combined filtrate was concentrated to give B-3 (44 g, 98%) as a solid.

[0452] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.85 (s, 1H), 4.71 (s, 1H), 2.48-2.20 (m, 3H), 2.12-1.98 (m, 3H), 1.90-1.49 (m, 10H), 1.47-1.08 (m, 8H), 1.01 (s, 3H), 0.99-0.71 (m, 2H), 0.58 (s, 3H).

Step 4

[0453] To a solution of B-3 (20 g, 63.5 mmol) in THF (300 mL) was added CsF (19.2 g, 127 mmol). To the mixture was added TMSCF.sub.3 (18.0 g, 127 mmol) dropwise at 10° C. The mixture was stirred at 10° C. for 2 hrs. To the mixture was added TBAF (127 mL, 1 M in THF, 127 mmol) at 10° C. The mixture was stirred at 20° C. for 3 hrs. To the mixture was added water (200 mL). The mixture was concentrated in vacuum to remove THF. To the residue was added EtOAc (300 mL). The organic layer was separated, washed with water (100 mL), brine (100 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum, triturated from PE:DCM (500 mL, 20:1), re-crystallized from MeCN (200 mL) to give B-4 (7.1 g) as a solid. The filtrate of trituration and re-crystallization was combined, concentrated in vacuum, purified by silica gel column (PE:EtOAc=30:1 to 10:1) twice to give impure B-4 which was re-crystallized from MeCN (200 mL) to give B-4 (7.6 g, total yield 60%) as a solid.

[0454] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.84 (s, 1H), 4.70 (s, 1H), 2.11-1.98 (m, 3H), 1.88-1.47 (m, 13H), 1.45-1.05 (m, 9H), 1.00-0.89 (m, 1H), 0.85 (s, 3H), 0.78-0.68 (m, 1H), 0.56 (s, 3H).

Step 5

[0455] To s solution of B-4 (14.7 g, 38.2 mmol) in THF (150 mL) was added 9-BBN dimer (10.7 g, 43.9 mmol). The mixture was stirred at 40° C. for 1 h. The mixture was cooled to 0° C. To the mixture was added EtOH (21.8 mL), NaOH (76.3 mL, 5 M, aq.) and H.sub.2O.sub.2 (38.1 mL, 10 M, aq.) dropwise. The mixture was stirred at 50° C. for 1 h. To the mixture was added Na.sub.2SO.sub.3 (200 mL, 25%, aq.) after cooled. The mixture was extracted with EtOAc (500 mL). The organic layer was separated, concentrated in vacuum and triturated form water (400 mL) to give B-5 (15 g, 98%) as a solid.

[0456] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.68-3.58 (m, 1H), 3.40-3.30 (m, 1H), 2.11-1.91 (m, 2H), 1.89-1.72 (m, 2H), 1.70-1.45 (m, 8H), 1.42-1.06 (m, 11H), 1.03 (d, J=6.4 Hz, 3H), 1.00-0.88 (m, 2H), 0.85 (s, 3H), 0.75-0.68 (m, 1H), 0.67 (s, 3H).

Step 6

[0457] To a solution of B-5 (15 g, 17.7 mmol) in DCM (60 mL) and pyridine (42 mL) was added TsCl (14.1 g, 74.4 mmol). The mixture was stirred at 15° C. for 2 hrs. To the mixture was added water (2 mL) and the mixture was stirred at 15° C. for 16 hrs. To the mixture was added water (100 mL). The mixture was extracted with PE/EtOAc (2:1, 300 mL). The organic layer was separated, washed with HCl (200 mL, 1 M), water (100 mL), brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give B-6 (23 g, crude) as a solid.

[0458] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.78 (d, J=8.0 Hz, 2H), 7.34 (d, J=8.0 Hz, 2H), 3.96 (dd, J=3.2, 9.2 Hz, 1H), 3.76 (dd, J=6.8, 9.2 Hz, 1H), 2.45 (s, 3H), 2.10-1.98 (m, 1H), 1.92-1.78 (m, 2H), 1.71-1.30 (m, 11H), 1.30-0.88 (m, 13H), 0.83 (s, 3H), 0.72-0.62 (m, 1H), 0.61 (s, 3H).

Step 7

[0459] To a solution of B-6 (23 g, 41.3 mmol) in DMF (100 mL) was added KI (27.3 g, 165 mmol). The mixture was stirred at 50° C. for 1 h. To the mixture was added PhSO.sub.2Na (20.1 g, 123 mmol). The mixture was stirred at 50° C. for 16 hrs. To the mixture was added DCM (200 mL), water (400 mL) and PE (2:1, 400 mL) with stirring. The organic layer was separated, washed with water (100 mL), brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to 150 mL in vacuum and a solid was formed. The mixture was filtered, washed with PE (100 mL), dried in vacuum to give B-7 (12 g, 55%) as a solid.

[0460] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.95-7.88 (m, 2H), 7.70-7.61 (m, 1H), 7.60-7.51 (m, 2H), 3.13 (d, J=13.2 Hz, 1H), 2.84 (dd, J=9.2, 14.0 Hz, 1H), 2.20-1.89 (m, 4H), 1.88-1.44 (m, 8H), 1.43-0.88 (m, 15H), 0.83 (s, 3H), 0.72-0.65 (m, 1H), 0.63 (s, 3H).

Step 8

[0461] To a solution of i-Pr.sub.2NH (573 mg, 5.67 mmol) in THF (10 mL) was added BuLi (1.88 mL, 2.5 M in hexane, 4.72 mmol) at −70° C. The mixture was warmed to 0° C. A solution of B-7 (1 g, 1.89 mmol) in THF (8 mL) was added at −70° C. The mixture was stirred at −70° C. for 1 h. To the mixture was added a solution of 2-isopropyloxirane (243 mg, 2.83 mmol) in THF (2 mL) at −70° C. The mixture was stirred at −70° C. for 1 h, 10° C. for 16 hrs and 50° C. for 2 hrs. To the mixture was added NH.sub.4Cl (5 mL, sat. aq.). The mixture was extracted with EtOAc (50 mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum and purified by silica gel column (PE:EtOAc=12:1 to 8:1) to give B-8 (0.5 g, 43%) as a solid.

[0462] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.95-7.85 (m, 2H), 7.70-7.52 (m, 3H), 3.63-3.46 (m, 1H), 3.44-3.31 (m, 1H), 2.18-1.61 (m, 8H), 1.55-1.11 (m, 13H), 1.11-0.78 (m, 18H), 0.72-0.60 (m, 2H), 0.50-0.40 (m, 3H).

Step 9

[0463] To a solution of B-8 (0.5 g, 0.815 mmol) in MeOH (50 mL) was added NiBr.sub.2 (2 mg, 0.009 mmol). Then magnesium powder (2.22 g, 91.4 mmol) was added in portions within 30 mins at 60° C. The mixture was stirred at 60° C. for 1 h. The mixture was poured into citric acid (200 mL, 10% aq.) and extracted with PE/EtOAc (2×200 mL, 1:1). The combined organic layer was washed with water (100 mL), brine (100 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum, purified by silica gel column (PE:EtOAc=20:1 to 10:1) to give Compound 2 (290 mg, 67%) as a solid.

[0464] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.37-3.28 (m, 1H), 2.12-2.01 (m, 1H), 2.00-1.91 (m, 2H), 1.87-1.77 (m, 2H), 1.71-1.58 (m, 5H), 1.50-1.00 (m, 19H), 0.96-0.88 (m, 10H), 0.85 (s, 3H), 0.72-0.67 (m, 1H), 0.67-0.64 (m, 3H). LCMS Rt=1.340 min in 2.0 min chromatography, 30-90 AB, No MS signal. HRMS ESI calcd. for C.sub.28H.sub.46F.sub.3O [M+H−H.sub.2O].sup.+ 455.3495, found 455.3489.

Step 10

[0465] Compound 2 (264 mg) was separated by silica gel column twice (300˜400 mesh, 30*250 mm, PE:EtOAc=30:1 to 15:1) to give Compound 2-A (56 mg, 21%) and Compound 2-B (101 mg, 38%) both as solids.

[0466] The diastereomeric ratio of 2-A and 2-B was assessed by conversion of the alcohol to a benzoate ester: To a solution of Compound 2-B (8 mg, 0.017 mmol) in DCM (0.5 mL) was added pyridine (132 mg, 1.68 mmol) and BzCl (23.7 mg, 0.169 mmol). The mixture was stirred at 25° C. for 20 mins. To the mixture was added PE (5 mL). The mixture was washed with NaHCO.sub.3 (2 mL, sat. aq.), HCl (2 mL, 1M, aq.), NaHCO.sub.3 (2 mL, sat. aq.), purified by prep-TLC (PE:DCM=1:1) to give 2-B-Bz for SFC analysis (98.7% de (“Column: Chiralpak AD-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO.sub.2 B: iso-propanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.”)).

[0467] To a solution of Compound 2-A (3 mg, 0.006 mmol) in DCM (0.5 mL) was added pyridine (50 mg, 0.633 mmol) and BzCl (8.9 mg, 0.063 mmol). The mixture was stirred at 25° C. for 20 mins. To the mixture was added PE (5 mL). The mixture was washed with NaHCO.sub.3 (2 mL, sat. aq.), HCl (2 mL, 1M, aq.), NaHCO.sub.3 (2 mL, sat. aq.), purified by prep-TLC (PE:DCM=1:1) to give 2-A-Bz for SFC analysis (95.0% d.e. (Column: Chiralpak AD-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO.sub.2 B: iso-propanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.)).

[0468] Compound 2-A:

[0469] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.37-3.28 (m, 1H), 2.12-2.01 (m, 1H), 2.00-1.91 (m, 2H), 1.87-1.77 (m, 2H), 1.71-1.58 (m, 5H), 1.50-1.00 (m, 19H), 0.96-0.88 (m, 10H), 0.85 (s, 3H), 0.72-0.67 (m, 1H), 0.65 (s, 3H). LCMS Rt=1.329 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.28H.sub.46F.sub.3O [M+H−H.sub.2O].sup.+ 455, found 455.

[0470] Compound 2-B:

[0471] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.37-3.28 (m, 1H), 2.12-2.01 (m, 1H), 2.00-1.91 (m, 2H), 1.87-1.77 (m, 2H), 1.71-1.58 (m, 4H), 1.50-1.30 (m, 10H), 1.30-1.00 (m, 10H), 0.96-0.88 (m, 10H), 0.85 (s, 3H), 0.72-0.67 (m, 1H), 0.66 (s, 3H). LCMS Rt=1.333 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.28H.sub.46F.sub.3O [M+H−H.sub.2O].sup.+ 455, found 455.

Synthesis of Compound 2-A—Absolute Stereochemistry

[0472] ##STR00088##

[0473] The experimental procedures of intermediate ST-200-CF3_6C can be found Example 5.

Synthesis of ST-200-096-001_1

[0474] ##STR00089##

[0475] To THF (1 mL) was added n-BuLi (0.948 mL, 2.5 M in hexane, 2.37 mmol), followed by adding a solution of ST-200-CF3_6C (500 mg, 0.949 mmol) in THF (4 mL) at −70° C. After stirring at −70° C. for 30 mins, (2R)-2-(propan-2-yl)oxirane (122 mg, 1.42 mmol) was added at −70° C. The mixture was warmed to 25° C. gradually and stirred at 25° C. for 16 hrs. The mixture was quenched with saturated NH.sub.4Cl (15 mL) and extracted with EtOAc (3×10 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered and concentrated to give ST-200-096-001_1 (560 mg, crude) as an oil, which was used directly for next step.

Synthesis of ST-200-096-001_2

[0476] ##STR00090##

[0477] To a solution of ST-200-096-001_1 (560 mg, 0.913 mmol) in methanol (30 mL) was added Mg powder (1.09 g, 45.6 mmol) under N.sub.2 at 65° C. The reaction mixture was quenched with HCl (60 mL) dropwise until the solution became clear. The reaction solution was extracted with EtOAc (3×30 mL). The combined organic layer was washed with sat. NaHCO.sub.3 (50 mL), brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-12% of EtOAc in PE) to give ST-200-096-001_2 (150 mg, 46%) as a solid.

[0478] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.35-3.26 (m, 1H), 2.10-1.91 (m, 3H), 1.88-1.76 (m, 2H), 1.71-1.62 (m, 4H), 1.52-1.35 (m, 6H), 1.32-1.20 (m, 7H), 1.17-0.98 (m, 6H), 0.95-0.87 (m, 10H), 0.86-0.80 (m, 4H), 0.72-0.61 (m, 4H).

Synthesis of ST-200-096-001_3

[0479] ##STR00091##

[0480] To a solution of ST-200-096-001_2 (200 mg, 0.423 mmol) in pyridine (3 mL) was added BzCl (177 mg, 1.26 mmol) at 0° C. and the reaction was stirred at 25° C. for 2 h. The reaction mixture was diluted with water (50 mL), extracted with EtOAc (2×40 mL). The organic layer was washed with brine (5×50 mL), dried over Na2SO4, filtered and concentrated. The crude was purified by silica gel column (PE/EtOAc=10/1 to 4/1) to give ST-200-096-001_3 (150 mg, 62%) as an oil.

[0481] The ST-200-096-001_3 (150 mg, 0.26 mmol) was separated by SFC (column: AD (250 mm*30 mm, 5 um)), gradient: 30-30% B (A=0.1% NH.sub.3H.sub.2O IPA)) to give ST-200-096-001_3 (120 mg, 81%) as a solid.

[0482] .sup.1HNMR (400 MHz, CDCl3) δ 8.05 (d, J=8 Hz, 2H), 7.55 (t, J=8 Hz, 1H), 7.44 (t, J=8 Hz, 2H), 4.98-4.91 (m, 1H), 2.09-1.89 (m, 4H), 1.86-1.61 (m, 6H), 1.53-1.34 (m, 8H), 1.27-1.03 (m, 8H), 0.99-0.95 (m, 8H), 0.92-0.83 (m, 7H), 0.71-0.61 (m, 4H).

[0483] SFC Rt=4.117 min in 10 min chromatography, AD_3_IPA_EtOH_5_40_25ML, 99% de.

Synthesis of Compound 2-A

[0484] ##STR00092##

[0485] To a solution of ST-200-096-001_3 (120 mg, 0.208 mmol) in THF (2 mL) and MeOH (1 mL) and water (1 mL) was added KOH (57.7 mg, 1.03 mmol). The mixture was stirred at 60° C. for 16 hrs, poured into water (20 mL) and extracted with EtOAc (2×40 mL). The combined organic layer was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (PE/EtOAc=5/l to 3/1) to give Compound 2-A (82 mg, 83%) as a solid.

[0486] .sup.1HNMR (400 MHz, CDCl3) δ 3.34-3.28 (m, 1H), 2.10-1.92 (m, 3H), 1.88-1.75 (m, 2H), 1.71-1.60 (m, 5H), 1.54-1.34 (m, 7H), 1.32-0.98 (m, 12H), 0.93-0.87 (m, 10H), 0.85 (s, 3H), 0.74-0.68 (m, 1H), 0.65 (s, 3H).

[0487] MS ESI calcd. For C.sub.28H.sub.47F.sub.3O.sub.2Na [M+Na.sup.+]=495, found 495.

Example 6. Synthesis of Compound 3

[0488] ##STR00093##

Step 1

[0489] To a solution of n-BuLi (568 μL, 2.5 M in hexane, 1.42 mmol) in THF (0.5 mL) at −65° C. under N.sub.2 was added a suspension of B-7 (300 mg, 0.5695 mmol) in THF (2.5 mL) drop-wise. The mixture was stirred for 30 minutes at −65° C. 2-(tert-butyl)oxirane (68.4 mg, 0.6834 mmol) was added drop-wise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually and stirred at 25° C. for 16 hours. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL), extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give C-1 (380 mg, crude) as a solid, which was used directly for the next step.

Step 2

[0490] To a solution of C-1 (380 mg, 0.6062 mmol) and NiCl.sub.2 (7.81 mg, 0.06062 mmol) in dry methanol (20 mL) was added Mg powder (580 mg, 24.2 mmol) in 4 portions under N.sub.2 with stirring at 50° C. The reaction mixture was stirred at 60° C. for 1 hour. The reaction mixture was cooled and poured into ethyl acetate (150 mL). The mixture was washed with 1 M HCl (3×200 mL), saturated NaHCO.sub.3 aqueous (200 mL), brine (200 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a solid, which was purified by silica gel chromatography (PE:EtOAc=8:1) to afford impure Compound 3 (310 mg) as a solid, which was purified by triturating in PE/DCM (15 mL/1 mL) to give Compound 3 (46 mg, 15%) as a solid.

[0491] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.16-3.05 (m, 1H), 2.09-2.01 (m, 1H), 2.01-1.92 (m, 2H), 1.89-1.76 (m, 2H), 1.73-1.60 (m, 3H), 1.52-1.33 (m, 8H), 1.32-0.93 (m, 12H), 0.93-0.87 (m, 12H), 0.85 (s, 4H), 0.73-0.61 (m, 4H).

[0492] .sup.19F NMR (400 MHz, CDCl.sub.3) δ 78.66.

[0493] LCMS Rt=1.354 min in 2 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.29H.sub.48F.sub.3O [M−H.sub.2O+H].sup.+ 469, found 469.

Example 7. Synthesis of Compound 4

[0494] ##STR00094##

Step 1

[0495] To a solution of diisopropylamine (0.2 mL) in THF (0.2 mL) was added butyllithium (0.57 mL, 2.5 M in n-hexane) at −70° C. The mixture was warmed to 25° C. and stirred at 25° C. for 30 minutes. The mixture was cooled to −70° C. and a solution of B-7 (250 mg, 16.5 mmol) in THF (3 mL) was added. After stirring at −70° C. for 1 h, (S)-3,3,3-trifluoro-2-hydroxy-2-methylpropyl 4-methylbenzenesulfonate, see Example 30. (169 mg, 0.57 mmol) was added at −70° C. The mixture was warmed to 25° C. and stirred at this temperature for 16 hours. The mixture was quenched with saturated aqueous NH.sub.4Cl (5 mL). The mixture was extracted with EtOAc (2×8 mL), washed with brine (2×20 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum to give a crude product D-1 (300 mg, crude) as an oil, which was used in the next step directly.

Step 2

[0496] To a solution of D-1 (300 mg, crude) in MeOH (15 mL) was added Mg powder (549 mg, 22.9 mmol) and NiCl.sub.2 (5 mg) at 60° C. The mixture was stirred at 60° C. for 1 h. EtOAc (20 mL) and aq. HCl (30 mL) was added. The mixture was extracted with EtOAc (2×30 mL). The combined organic layers were washed with water (3×50 mL), sat. NaHCO.sub.3 (2×50 mL), brine (2×50 mL) to give a crude product, which was purified by flash column (0-30% of EtOAc in PE) to give Compound 4 (100 mg, impure), which was triturated with CH.sub.3CN (5 mL) at 25° C. to give Compound 4 (50 mg, 50%) as a solid.

[0497] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.10-1.90 (m, 3H), 1.85-1.75 (m, 3H), 1.70-1.60 (m, 5H), 1.50-1.30 (m, 6H), 1.25-1.00 (m, 14H), 0.90-0.80 (m, 7H), 0.70-0.55 (m, 4H).

[0498] LCMS Rt=1.264 min in 2 min chromatography, 30-90 AB, MS ESI calcd. For C.sub.27H.sub.41F.sub.6O [M+H−H.sub.2O].sup.+ 495, found 495.

Example 8. Synthesis of Compound E-1

[0499] ##STR00095##

Step 1

[0500] To a solution of S,S-cat (2 g, 3.65 mmol) in anhydrous DCM (30 mL) was added a solution of cobalt(II) acetate (775 mg, 4.38 mmol) in MeOH (30 mL) under nitrogen at 20° C. The mixture was stirred for 30 mins at 20° C. and at 0° C. for 1 h. The precipitated solid was filtered, washed with cold MeOH (2×30 mL) and dried in vacuum to give Co-S,S-cat (1.6 g, 73%) as a solid.

Step 2

[0501] To a solution of Co-S,S-cat (1.07 g, 1.78 mmol) in toluene (30 mL) was added AcOH (1.12 g, 18.7 mmol). The mixture was stirred at 20° C. for 30 mins. The solution was concentrated in vacuum to give a solid. The resulting catalyst residue was dissolved in neat E-0 (100 g, 892 mmol) at 20° C., the reaction mixture was cooled to 0° C., and water (8.82 g, 490 mmol) was added dropwise. The mixture was warmed to 20° C. and stirred for 48 hrs. E-1 (44 g) was isolated by distillation from the reaction mixture.

[0502] .sup.1H NMR (400 MHz, DMSO-d6) δ 3.96 (s, 1H), 3.11-2.98 (m, 2H).

[0503] The e.e. of E-1 was determined by opening the epoxide with benzylamine. E-1 (200 mg, 1.78 mmol) was added to dry benzylamine (190 mg, 1.78 mmol), and the mixture was stirred at 20° C. for 2 hrs. A solid precipitated, which was triturated from petroleum ether to afford the product (260 mg, 67%) as a solid. The e.e. of this product was determined to be 100% by chiral HPLC (Column: CD-PH 250*4.6 mm I.D., 5 um; Mobile phase: from 10% to 80% of B in A (A: Water with 0.069% TFA B: Acetonitrile); Flow rate: 0.8 mL/min; Column Temperature: 30° C.).

Example 9. Synthesis of Compound 5

[0504] ##STR00096##

Step 1

[0505] To a solution of n-BuLi (0.704 mL, 2.5 M in hexane, 1.76 mmol) in THF (0.5 mL) at −65° C. under N2 was added a suspension of B-7 (310 mg, 0.588 mmol) in THF (2.5 mL) dropwise and the reaction was stirred for 30 minutes at −65° C. A solution of E-1 (78.9 mg, 0.705 mmol) was added dropwise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually and stirred at 25° C. for 16 hours. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL), extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give E-2 (300 mg, crude) as a solid, which was used directly for the next step.

Step 2

[0506] To a solution of E-2 (300 mg, 0.469 mmol) and nickel (II) chloride (15.1 mg, 0.117 mmol) in dry methanol (20 mL) was added magnesium powder (454 mg, 18.7 mmol) under N.sub.2 with stirring at 50° C. to initiate continuous hydrogen generation. The reaction mixture was stirred at 60° C. for 1 hour. The reaction mixture was quenched by 2M HCl (100 mL) dropwise at 10° C. until the solid was dissolved. After extracting with EtOAc (2×150 mL), the combined organic layer was washed with sat. NaHCO.sub.3 aq. (300 mL), brine (300 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a solid, which was purified by silica gel chromatography (PE:THF=12:1) to give the product. The residue was re-crystallized from MeCN (10 mL) to afford Compound 5 (41 mg, 18%) as a solid.

[0507] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.75-3.65 (m, 1H), 2.10-1.95 (m, 3H), 1.90-1.75 (m, 2H), 1.73-1.66 (m, 5H), 1.56-1.30 (m, 14H), 1.29-1.01 (m, 5H), 1.00-0.85 (m, 3H), 0.84 (s, 3H), 0.67-0.60 (m, 4H).

[0508] LCMS Rt=1.226 min in 2.0 min chromatography, 30-90 AB.

Example 10. Synthesis of Compound 6

[0509] ##STR00097##

Step 1

[0510] To a solution of n-BuLi (0.568 mL, 2.5 M in hexane, 1.42 mmol) in THF (0.5 mL) at −65° C. under N.sub.2 was added a suspension of B-7 (250 mg, 0.474 mmol) in THF (2.5 mL) dropwise. After stirring at −65° C. for 30 minutes, a solution of (2S)-2-methyloxirane (32.9 mg, 0.568 mmol) was added dropwise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually and stirred at 25° C. for 16 hours. The reaction mixture was quenched with saturated NH.sub.4Cl aqueous (30 mL), and extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give F-1 (250 mg, crude) as a solid, which was used directly for the next step.

Step 2

[0511] To a solution of F-1 (250 mg, 0.427 mmol) and nickel (II) chloride (13.7 mg, 0.106 mmol) in dry methanol (20 mL) was added magnesium powder (413 mg, 17.0 mmol) under N.sub.2 with stirring at 50° C. to initiate continuous hydrogen generation. The reaction mixture was stirred at 60° C. for 1 hour. The reaction mixture was quenched by 2M HCl (100 mL) which was added dropwise at 10° C. until the solid was dissolved. After extracting with EtOAc (2×150 mL), the combined organic layer was washed with sat. NaHCO.sub.3 aq. (300 mL), brine (300 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a solid, which was purified by silica gel chromatography (PE/THF=12/1) to give impure Compound 6 (100 mg, containing 12% of 22,23-olefin by NMR) as a solid. To a solution of the impure Compound 6 (100 mg, 0.224 mmol) in EtOAc (10 mL) was added Pd/C (26.5 mg, 0.224 mmol) under N.sub.2 to remove the undesired olefin. The mixture was degassed under vacuum and purged with H.sub.2 several times. The mixture was stirred for 2 hrs at 25° C. under H.sub.2. The mixture was filtered and the filtrate was concentrated in vacuum to give residue. The residue was purified by re-crystallization from MeCN (10 mL) to give Compound 6 (35 mg, 19%) as a solid.

[0512] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.75-3.65 (m, 1H), 2.10-1.95 (m, 3H), 1.90-1.75 (m, 2H), 1.73-1.66 (m, 4H), 1.56-1.30 (m, 8H), 1.29-1.01 (m, 14H), 1.00-0.85 (m, 4H), 0.84 (s, 3H), 0.67-0.60 (m, 4H).

[0513] LCMS Rt=1.222 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.28H.sub.42F.sub.3O [M+H−H.sub.2O].sup.− 427, found 427.

Example 11. Syntheses of Compounds 7, 7-A, and 7-B

[0514] ##STR00098## ##STR00099##

[0515] X-ray data of Compound 7 confirmed stereochemistry of Compound 7-A and Compound 7-B.

Step 1

[0516] To a solution of compound G-1 (5.0 g, 12.8 mmol) in EtOAc (150 mL) was added Pd/C (1.0 g), then the mixture was stirred under hydrogen (50 psi) at 50° C. overnight. The mixture was filtered through a pad of celite and the filtrate was evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: petroleum ether:ethyl acetate=15:1) to afford the pure product G-2 (3.7 g, 74%).

[0517] .sup.1H NMR: (400 MHz, CDCl.sub.3) δ 3.66 (s, 3H), 3.53-3.62 (m, 1H), 2.40-2.30 (m, 1H), 2.26-2.18 (m, 1H), 1.97-1.62 (m, 6H), 1.60-1.20 (m, 13H), 1.18-0.93 (m, 6H), 0.92 (d, J=6.8 Hz, 3H), 0.90-0.82 (m, 1H), 0.79 (s, 3H), 0.64-0.59 (m, 4H).

Step 2

[0518] To a solution of G-2 (10 g, 25.6 mmol) in DCM (200 mL) was added DMP (19.5 g, 46 mmol) at 25° C. The mixture was stirred at 25° C. for 30 min. Water (80 mL) was added followed by NaHCO.sub.3 (20 g) and the mixture was filtered. The filtrate was extracted with DCM (100 mL), washed with Na.sub.2SO.sub.3 (2×300 mL) and brine (2×300 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum to give a crude product G-3 (9 g) as a solid, which was used in the next step without further purification.

[0519] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.66 (s, 3H), 2.41-2.29 (m, 1H), 2.27-2.16 (m, 1H), 2.10-1.91 (m, 3H), 1.88-1.62 (m, 6H), 1.52-0.98 (m, 16H), 0.97-0.87 (m, 4H), 0.84 (s, 3H), 0.73-0.63 (m, 4H).

Step 3

[0520] To a mixture of G-3 (7 g, 18.0 mmol) and CsF (5.46 g, 36.0 mmol) in THF (70 mL) was added drop wise TMSCF.sub.3 (5.11 g, 36.0 mmol) at 0° C. The mixture was stirred and kept below 10° C. for 10 min. TBAF (45.0 mL, 1 M in THF, 45.0 mmol) was added at 10° C. and the mixture was stirred and kept below 10° C. for 10 min. After that, the mixture was treated with water (200 mL) and extracted with EtOAc (2×200 mL). The combined organic layers was washed with brine (500 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue was purified by silica gel chromatograph (PE/EtOAc=5/1) to afford G-4 (5.55 g, 67%), 4H), 0.84 (s, 3H), 0.73-0.63 (m, 4H).

Step 4

[0521] To a suspension of LiAlH.sub.4 (1.03 g, 27.4 mmol) in THF (80 mL) was added a solution of G-4 (6.3 g, 13.7 mmol) in THF (20 mL) under N.sub.2 dropwise at 0° C. The reaction was stirred at 25° C. for 2 h. The reaction was quenched with water/THF (1/10, 40 mL) followed by adding 2 M HCl (100 mL) at 0° C. The mixture was extracted with EtOAc (2×100 mL). The combined organic phase was washed with brine (300 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to afford G-5 (5 g, crude) as a solid.

[0522] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.61 (s, 2H), 2.11-1.92 (m, 4H), 1.90-1.77 (m, 2H), 1.73-1.60 (m, 5H), 1.52-0.98 (m, 17H), 0.96-0.87 (m, 4H), 0.85 (s, 3H), 0.73-0.64 (m, 4H).

Step 5

[0523] To a solution of G-5 (3 g, 6.96 mmol) in DCM (30 mL) was added DMP (5.89 g, 13.9 mmol) at 20° C. The reaction mixture was stirred at 20° C. for 20 min and quenched with saturated NaHCO.sub.3 aqueous (30 mL) at 20° C. The mixture was filtered. The DCM layer was separated and the aqueous phase was extracted with DCM (30 mL). The combined organic phase was washed with saturated Na.sub.2SO.sub.3 aqueous (3×50 mL), brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum, the residue was triturated from CH.sub.3CN (5 mL) at 20° C. to give G-6 (1.3 g, 44%) as a solid.

[0524] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.78-9.75 (t, J=2.00 Hz, 1H), 2.51-2.20 (m, 2H), 2.11-1.74 (m, 6H), 1.74-0.97 (m, 19H), 0.96-0.87 (m, 4H), 0.85 (s, 3H), 0.73-0.67 (m, 1H), 0.65 (s, 3H).

Step 6

[0525] To a suspension of Mg (2 g, 82.2 mmol) and I.sub.2 (10 mg) in THF (2 mL) was added a solution of bromocyclobutane (5 g, 37.0 mmol) in THF (8 mL) at 60° C. dropwise. The mixture was stirred at 60° C. for 1 h. The mixture was diluted with THF (10 mL) and used directly. The Grignard reagent was added to a solution of G-6 (0.6 g, 1.40 mmol) in THF (5 mL) at 0° C. The mixture was stirred at 0° C. for 1 h and quenched with NH.sub.4Cl (10 mL, sat. aq.). The mixture was extracted with EtOAc (3×20 mL). The organic layer was separated, concentrated in vacuum, purified by silica gel (PE/EtOAc=20/l to 5/1) to give a crude product, which was re-crystallized from MeCN/H.sub.2O (5/2, 15 mL) to give Compound 7 (250 mg, 37%) as a solid.

[0526] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.49-3.38 (m, 1H), 2.40-2.25 (m, 1H), 2.10-1.90 (m, 5H), 1.90-1.60 (m, 9H), 1.57-1.18 (m, 14H), 1.17-0.96 (m, 6H), 0.96-0.86 (m, 4H), 0.84 (s, 3H), 0.73-0.62 (m, 4H).

[0527] HPLC Rt=6.10 min in 8.0 min chromatography, 50-100 AB.

[0528] MS ESI calcd. for C.sub.29H.sub.46F.sub.3O [M+H−H.sub.2O].sup.+ 467, found 467.

Step 7

[0529] To a solution of Compound 7 (200 mg, 0.412 mmol) in DCM (5 mL) was added pyridine (650 mg, 8.23 mmol) and BzCl (347 mg, 2.47 mmol). The mixture was stirred at 25° C. for 1 h. The mixture was treated with H.sub.2O (5 mL) and washed with HCl (10 mL, 1 M, aq.), NaHCO.sub.3 (10 mL, sat. aq.), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum to give a crude product, which was purified by silica gel column (PE/EtOAc=10/l) to give 300 mg of impure product. The impure product was separated by SFC (column: Chiralpak AD-3 50*4.6 mm I.D., 3 um); Condition: Base-IPA; Gradient: 5-40% B; flow rate: 4 mL/min) to give G-6-A (75 mg, 31%, t.sub.R=5.282 min, 100% d.e. (“Column: Chiralpak AD-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO.sub.2 B: iso-propanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.”)) and G-6-B (88 mg, 36%, t.sub.R=4.827 min, 100% d.e. (“Column: Chiralpak AD-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO.sub.2 B: iso-propanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.”)).

Step 8a

[0530] To a solution of G-7-A (75 mg, 0.127 mmol) in THF (5 mL) and MeOH (1 mL) was added a suspension of LiOH.H.sub.2O (399 mg, 9.52 mmol) in water (1 mL). The mixture was stirred at 60° C. for 24 h. After removing the organic solvent in vacuum, the mixture was treated with H.sub.2O (5 mL) and extracted with EtOAc (3×5 mL). The organic layers were washed with brine (2×15 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum. The residue was triturated from CH.sub.3CN (2 mL) at 25° C. to give Compound 7-A (43 mg, 70%) as a solid.

[0531] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.49-3.41 (m, 1H), 2.37-2.26 (m, 1H), 2.10-1.75 (m, 10H), 1.75-1.60 (m, 4H), 1.52-1.15 (m, 16H), 1.15-0.93 (m, 4H), 0.92-0.82 (m, 7H), 0.73-0.62 (m, 4H).

[0532] HPLC Rt=6.78 min in 8.0 min chromatography, 30-90 AB.

[0533] MS ESI calcd. for C.sub.29H.sub.46F.sub.3O [M+H−H.sub.2O].sup.+ 467, found 467.

Step 8b

[0534] To a solution of G-7-B (88 mg, 0.149 mmol) in THF (5 mL) and MeOH (1 mL) was added a suspension of LiOH.H.sub.2O (406 mg, 9.68 mmol) in water (1 mL). The mixture was stirred at 60° C. for 24 h. After removing the organic solvent in vacuum, the mixture was treated with H.sub.2O (5 mL) and extracted with EtOAc (3×5 mL). The organic layers were washed with brine (2×15 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum. The residue was triturated from CH.sub.3CN (2 mL) at 25° C. to give Compound 7-B (52 mg, 72%) as a solid.

[0535] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.48-3.37 (m, 1H), 2.39-2.26 (m, 1H), 2.10-1.74 (m, 10H), 1.72-1.61 (m, 4H), 1.53-1.19 (m, 13H), 1.19-0.94 (m, 7H), 0.94-0.80 (m, 7H), 0.73-0.62 (m, 4H). HPLC Rt=6.78 min in 8.0 min chromatography, 30-90 AB MS ESI calcd. for C.sub.29H.sub.46F.sub.3O [M+H−H.sub.2O].sup.+ 467.3495, found 467.3.

Example 12. Synthesis of Compound H-1

[0536] ##STR00100##

[0537] To a suspension of Me.sub.3SI (3.93 g, 19.3 mmol) in THF (20 mL) was added a solution of t-BuOK (3.33 g, 29.8 mmol) in THF (10 mL) under N.sub.2 at 15° C. The suspension was stirred at 15° C. for 30 mins. A solution of H-0 (2 g, 14.9 mmol) in THF (5 mL) was added dropwise at 15° C. The mixture was stirred at 15° C. for 16 hrs. The mixture was quenched with Sat. NH.sub.4Cl (50 mL) and extracted with EtOAc (3×20 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, and concentrated to give H-1 (1.8 g, 82%) as a solid.

[0538] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.72 (s, 2H), 2.20-1.85 (m, 8H).

Example 13. Synthesis of Compound 8

[0539] ##STR00101##

Step 1

[0540] To a solution of THF (5 mL) and BuLi (3.78 mL, 2.5 M in hexane, 9.47 mmol) was added a solution of B-7 (2 g, 3.79 mmol) in THF (15 mL) at −70° C. After stirring at −70° C. for 1 h, a solution of H-1 (1.68 g, 5.68 mmol) in THF (5 mL) was added at −70° C. The mixture was stirred at −70° C. for another 1 h. The mixture was warmed to 25° C. and stirred for 16 hrs and quenched by adding NH.sub.4Cl (50 mL, sat. aq.). The mixture was extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, concentrated, and purified by combi-flash (0-10% of EtOAc in PE) to give H-2 (250 mg, 10%) as a solid and 1.8 g of starting material which was recycled.

[0541] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.00-7.92 (m, 2H), 7.73-7.65 (m, 1H), 7.63-7.52 (m, 2H), 3.62-3.55 (m, 1H), 2.37-2.28 (m, 1H), 2.15-1.94 (m, 4H), 1.94-1.85 (m, 6H), 1.85-1.55 (m, 5H), 1.55-1.43 (m, 6H), 1.43-1.10 (m, 10H), 1.10-0.90 (m, 3H), 0.90-0.70 (m, 6H), 0.70-0.57 (m, 1H), 0.55 (s, 3H).

Step 2

[0542] To a solution of H-2 (250 mg, 0.37 mmol) in MeOH (15 mL) was added Mg powder (355 mg, 14.8 mmol) at 55° C. The mixture was stirred at 60° C. for 16 hrs. The mixture was quenched with HCl (50 mL, 1N) until the reaction became clear and extracted with DCM (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give Compound 8 (55 mg, 28%) as a solid.

[0543] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.20-1.73 (m, 9H), 1.73-1.58 (m, 7H), 1.58-0.85 (m, 11H), 0.85-1.00 (m, 8H), 1.00-0.86 (m, 5H), 0.85 (s, 3H), 0.72-0.62 (m, 4H).

[0544] LCMS Rt=1.286 min in 2 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.30H.sub.46F.sub.5O [M−H.sub.2O+H].sup.+ 517, found 517.

Example 14. Synthesis of Compound 9

[0545] ##STR00102##

[0546] To a suspension of Mg (1.37 g, 56.5 mmol) and I.sub.2 (10 mg) in THF (2 mL) was added a solution of 4-chlorotetrahydro-2H-pyran (2.72 g, 22.6 mmol) in THF (8 mL) at 60° C. dropwise. The mixture was stirred at 60° C. for 2 h. The mixture was diluted with THF (10 mL) and used directly. The Grignard reagent was added to a solution of G-6 (0.55 g, 1.28 mmol) in THF (5 mL) at 0° C. The mixture was stirred at 0° C. for 1 h and treated with NH.sub.4Cl (10 mL, sat. aq.). The mixture was extracted with EtOAc (3×20 mL). The organic layer was separated, concentrated in vacuum, purified by silica gel column (PE/EtOAc=20/l to 5/1) to give a crude product, which was re-crystallized from CH.sub.3CN (10 mL) to give Compound 9 (180 mg, 27%) as a solid.

[0547] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.05-3.97 (m, 2H), 3.41-3.25 (m, 3H), 2.10-1.91 (m, 3H), 1.88-1.57 (m, 7H), 1.55-1.33 (m, 11H), 1.33-0.96 (m, 12H), 0.96-0.86 (m, 4H), 0.85 (s, 3H), 0.72-0.63 (m, 4H).

[0548] HPLC Rt=4.73 min in 8.0 min chromatography, 50-100 AB.

[0549] MS ESI calcd. for C.sub.30H.sub.48F.sub.3O.sub.2 [M+H−H.sub.2O].sup.+ 497, found 497.

Example 15. Synthesis of Compound J-1

[0550] ##STR00103##

[0551] To a mixture of trimethylsulfoxonium iodide (30.6 g, 150 mmol) in THF (100 mL) was added NaH (5.98 g, 60% in mineral oil, 150 mmol) in portions at 0° C. under N.sub.2. The mixture was stirred at 0° C. for 30 mins. Dihydrofuran-3(2H)-one (10 g, 116 mmol) in DMSO (100 mL) was added dropwise at 0° C. The reaction mixture was stirred at 0° C. for 2 hours. The mixture was poured into ice-water (500 mL) in portions, extracted with DCM (2×500 mL). The combined organic phase was washed with brine (500 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated at 30° C. The residue was purified by Combi-flash (EtOAc in PE, 0%˜40%) to afford J-1 (1.5 g, 13%) as an oil.

[0552] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.11-3.90 (m, 3H), 3.66 (d, J=10.0 Hz, 1H), 3.03 (d, J=4.4 Hz, 1H), 2.94 (d, J=4.0 Hz, 1H), 2.34-2.23 (m, 1H), 2.00-1.88 (m, 1H).

Example 16. Synthesis of Compound 10

[0553] ##STR00104##

Step 1

[0554] To a solution of n-BuLi (0.95 mL, 2.38 mmol, 2.5 M) in THF (2 mL) under N.sub.2 at −70° C. was added a suspension of A-7 (see Example 3) (500 mg, 0.95 mmol) in THF (5 mL) drop-wise to give a suspension. After stirring at −70° C. for 30 min, a solution of J-1 (238 mg, 2.38 mmol) in THF (3 mL) was added. Then the reaction was stirred at −70° C. for 10 min and 20° C. for 16 hours. The reaction was quenched with sat. NH.sub.4Cl (20 mL), extracted with EtOAc (3×20 mL). The combined organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product J-2 (500 mg) as a solid, which was used directly in next step.

[0555] LCMS Rt=0.925 min in 1.5 min chromatography, 5-95 AB, MS ESI calcd. for C.sub.34H.sub.47F.sub.3O.sub.5SNa [M+Na].sup.+ 647, found 647.

Step 2

[0556] To a solution of J-2 (300 mg, 0.48 mmol) in 20 mL of dry methanol under N.sub.2 was added magnesium turnings (466 mg, 19.2 mmol) (activated with 0.5% aqueous HCl, water, dry ethanol, and MTBE) and NiCl.sub.2 (12.4 mg, 0.96 mmol) with stirring at 55° C. to initiate continuous hydrogen generation. After the addition of a further two batches of 466 mg of magnesium turnings, most of the starting material was consumed. The reaction mixture was quenched by 2M HCl (100 mL) which was added dropwise at 10° C. until solid was dissolved. After extraction with DCM (3×80 mL), the combined organic phase was washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by Combi-flash (0%˜50% of EtOAc in PE) to afford Compound 10 (46 mg, 20%) as a solid.

[0557] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.43-5.32 (m, 1H), 4.08-3.98 (m, 1H), 3.95-3.85 (m, 1H), 3.75-3.66 (m, 1H), 3.59-3.51 (m, 1H), 2.53-2.45 (m, 2H), 2.11-1.87 (m, 6H), 1.82-1.65 (m, 4H), 1.54-1.38 (m, 7H), 1.33-1.12 (m, 6H), 1.08-0.92 (m, 9H), 0.79-0.61 (m, 4H).

[0558] LCMS Rt=1.121 min in 2 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.30H.sub.46F.sub.3O.sub.3NNa [M+MeCN+Na].sup.+ 548, found 548.

Example 17. Synthesis of Compound 11

[0559] ##STR00105##

Step 1

[0560] To a solution of n-BuLi (452 μL, 2.5 M in hexane, 1.13 mmol) in THF (0.5 mL) at −65° C. under N.sub.2 was added a suspension of B-7 (200 mg, 0.3797 mmol) in THF (2.5 mL) was added drop-wise and stirred for 30 minutes at −65° C. After that, diisopropylamine (114 mg, 1.13 mmol) was added at −65° C., followed by adding 1,6-dioxaspiro[2.5]octane (65.0 mg, 0.5695 mmol) was added drop-wise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually and stirred at 25° C. for 16 hour. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL), extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give K-1 (380 mg, crude) as a solid, which was used directly for the next step.

Step 2

[0561] To a solution of K-1 (0.348 g, 0.543 mmol) in MeOH (20 mL) was added Mg (0.520 g, 21.7 mmol) and NiCl.sub.2 (3.51 mg, 0.0271 mmol) at 60° C. The mixture was stirred at 60° C. for 1 hour. The reaction mixture was cooled to 25° C. The mixture was added in HCl (20 mL, 1 M in water). The mixture was extracted with EtOAc (2×20 mL), washed with NaHCO.sub.3 (2×40 mL) and brine (2×40 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum. The crude residue was purified by silica gel column (PE/EtOAc=10/l to 2/1) to give 66 mg of impure Compound 11 as a solid, which was triturated from CH.sub.3CN (5 mL) at 25° C. to give Compound 11 (30 mg, 11%) as a solid.

[0562] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.84-3.64 (m, 4H), 2.11-1.90 (m, 3H), 1.87-1.61 (m, 6H), 1.51-1.20 (m, 16H), 1.18-0.96 (m, 7H), 0.94-0.80 (m, 7H), 0.74-0.61 (m, 4H).

[0563] LCMS Rt=1.170 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.29H.sub.46F.sub.3O.sub.2 [M+H−H.sub.2O].sup.+ 483, found 483.

Example 18: Synthesis of Compound 1839

[0564] ##STR00106##

[0565] The experimental of intermediate ST-200-INT_2, or A2, can be found in Example 3.

Synthesis of ST-200-CF3_1A

[0566] ##STR00107##

[0567] A solution of ST-200-INT_2 (9.5 g, 30.4 mmol) and TMSCF.sub.3 (12.9 g, 91.2 mmol) in THF (50 mL) was added dropwise within 30 mins at 0° C. to a suspension of CsF (462 mg, 3.04 mmol) in THF (100 mL). The mixture was stirred at 10° C. for 16 hrs. TLC showed the starting material remained. The mixture was cooled to 0° C. TBAF (3 mL, 1 M in THF, 3 mmol, Aldrich) was added to the mixture at 0° C. The mixture was stirred at 10° C. for 1 h. TBAF (91.2 mL, 1 M in THF, 91.2 mmol) was added to the mixture. The mixture was stirred at 10° C. for another 1 h. The mixture was concentrated in vacuum. The residue was dissolved in EtOAc (100 mL), washed with water (3×100 mL) and concentrated in vacuum to yield a crude product, which was combined with another batch of 9.5 g ST-200-INT_2, purified by silica gel column (PE:EtOAc=30:1 to 20:1) in four parts to give ST-200-CF3_1B (2.3 g, purity 83%, yield 8%) and ST-200-CF3_1A (6.2 g, purity 32%, yield 8%). 3.0 g of impure ST-200-CF3_1A was used in the step directly and another 3.2 g was purified by silica gel column (PE:EtOAc=30:1 to 20:1) and re-crystallized form MeCN (10 mL) to give ST-200-CF3_1A (0.5 g, purity 94%).

[0568] Note: ST-200-CF3_1A and ST-200-CF3_1B were identified from .sup.3J.sub.H,CF, (FDCS). (J. Org. Chem. 2015, 80, 1754)

[0569] ST-200-CF3_1A:

[0570] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.43-5.33 (m, 1H), 4.85 (s, 1H); 4.71 (s, 1H); 2.49 (s, 2H); 2.11-1.97 (m, 4H), 1.95-1.32 (m, 14H), 1.30-0.98 (m, 7H), 0.59 (s, 3H).

[0571] ST-200-CF3_1B:

[0572] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.54-5.41 (m, 1H), 4.86 (s, 1H); 4.72 (s, 1H); 2.78-2.65 (m, 1H); 2.18-1.97 (m, 3H), 1.95-1.35 (m, 16H), 1.32-0.98 (m, 7H), 0.59 (s, 3H).

Synthesis of ST-200-CF3_2A

[0573] ##STR00108##

[0574] 9-BBN dimer (2.19 g, 9.01 mmol) was added to a solution of ST-200-CF3_1A (3 g, impure) in THF (35 mL). The mixture was stirred at 40° C. for 1 h. Next, EtOH (4.5 mL), NaOH (15.6 mL, 5 M, aq.) and H.sub.2O.sub.2 (7.83 mL, 10 M, aq.) were added dropwise and the mixture was cooled to 0° C. The mixture was stirred at 50° C. for 1 h. Na.sub.2SO.sub.3 (100 mL, 10%, aq.) was added to the mixture after cooling. The mixture was extracted with EtOAc (100 mL). The organic layer was separated, purified by silica gel column (PE:EtOAc=10:1 to 7:1) to give ST-200-CF3_2A (1.2 g, purity 79%, yield 30%) as a solid.

[0575] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.32 (m, 1H), 3.64 (dd, J=2.8, 10.4 Hz, 1H), 3.36 (dd, J=6.8, 10.4 Hz, 1H), 2.50 (s, 2H), 2.32-1.92 (m, 4H), 1.92-1.70 (m, 4H), 1.70-1.29 (m, 8H), 1.29-0.91 (m, 11H), 0.71 (s, 3H).

Synthesis of ST-200-CF3_3A

[0576] ##STR00109##

[0577] TsCl (1.14 g, 5.98 mmol) was added to a solution of ST-200-CF32A (1.2 g, 2.99 mmol) in DCM (5 mL) and py (3.5 mL). The mixture was stirred at 15° C. for 2 hrs. PE (10 mL) was added to the mixture. The mixture was washed with water (10 mL) and brine (10 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum and purified by silica gel column (PE:DCM:EtOAc=5:1:0.3 to 5:1:0.4) to give ST-200-CF3_3A (1.05 g, 64%) as a solid.

[0578] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.78 (d, J=8.4 Hz, 2H), 7.34 (d, J=8.4 Hz, 2H), 5.40-5.33 (m, 1H), 3.97 (dd, J=2.8, 9.2 Hz, 1H), 3.77 (dd, J=6.4, 9.2 Hz, 1H), 2.48 (s, 2H), 2.45 (s, 3H), 2.10-1.88 (m, 5H), 1.82-1.35 (m, 9H), 1.30-0.82 (m, 12H), 0.64 (s, 3H).

Synthesis of ST-200-CF3_4A

[0579] ##STR00110##

[0580] KI (1.25 g, 7.56 mmol) was added to a solution of ST-200-CF3_3A (1.05 g, 1.89 mmol) in DMF (5 mL). The mixture was stirred at 50° C. for 1 h. To the mixture was added PhSO.sub.2Na (0.93 g, 5.67 mmol). The mixture was stirred at 50° C. for 2 hrs. Water (10 mL) and DCM (30 mL) were added to the mixture. The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum and triturated from PE/DCM (10 mL, 5:1) to give ST-200-CF3_4A (600 mg, 61%) as a solid.

[0581] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.98-7.87 (m, 2H), 7.70-7.52 (m, 3H), 5.39-5.31 (m, 1H), 3.14 (d, J=14.4 Hz, 1H), 2.85 (dd, J=9.6, 14.0 Hz, 1H), 2.48 (s, 2H), 2.20-1.88 (m, 5H), 1.88-1.68 (m, 4H), 1.60-1.33 (m, 5H), 1.30-0.82 (m, 12H), 0.64 (s, 3H).

Synthesis of E-322_6_1

[0582] ##STR00111##

[0583] Diisopropylamine (3.76 mmol, 380 mg) was added to THF (2 mL) under N.sub.2 at −70° C., followed by an addition of n-BuLi (3.42 mmol, 1.36 mL, 2.5M in hexane 3.0 eq). The reaction was allowed to warm to 15° C. and was then re-cooled to −70° C. A suspension of ST-200-CF3_4A (1.14 mmol, 600 mg) in THF (5 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, a solution of 2,2-dimethyloxirane (2.28 mmol, 218 mg, 2.0 eq.) in THF (1 mL) was added over 5 min (slightly exothermic, keeping internal T<−70° C.). Then reaction was stirred at 15° C. for 12 hrs. The reaction was quenched with sat. NH.sub.4Cl (30 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated to give ST-200-CF3_5A (600 mg, crude) as a foam.

Synthesis of 1839

[0584] ##STR00112##

[0585] Mg powder (960 mg, 40 mmol) was added to a solution of E-322_6_1 (600 mg, 1 mmol) in MeOH (10 mL) at 55° C. The reaction mixture was stirred at 60° C. under N.sub.2 for 2 hrs. The mixture was quenched with HCl (100 mL, 2 M) until the reaction became clear and extracted with DCM (3×20 mL). The combined organic phase was washed with sat. NaHCO.sub.3 (50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-10% of EtOAc in PE) to give 170 mg impure product, which was purified again by prep-HPLC (column: DuraShell 150*25 mm*5 um), gradient: 75-100% B (A=0.05% HCl/H.sub.2Q. B=MeCN), flow rate: 30 mL/min) to give 1839 (66 mg, 14%) as a solid.

[0586] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.37-5.36 (m, 1H), 2.48 (s, 2H), 2.10-1.92 (m, 4H), 1.90-1.70 (m, 3H), 1.62-1.58 (m, 2H), 1.56-1.35 (m, 7H), 1.34-1.22 (m, 3H), 1.21-1.07 (m, 10H), 1.06 (s, 3H), 1.05-0.98 (m, 2H), 0.93 (d, J=6.8 Hz, 3H), 0.68 (s, 3H).

[0587] LCMS Rt=1.277 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.27H.sub.42F.sub.3O [M+H−H.sub.2O].sup.+ 439, found 439.

Example 19: Synthesis of 1967

[0588] ##STR00113##

[0589] The synthesis of ST-200-CF3_6C or B7 can be found in Example 5.

Synthesis of 200-DA-C24_8_2

[0590] ##STR00114##

[0591] Sodium hydride (18.0 g, 60% in mineral oil, 452 mmol) was added in portions to a mixture of trimethylsulfoxonium iodide (92.2 g, 452 mmol) in THF (300 mL) at 0° C. under N.sub.2. The mixture was stirred at 0° C. for 30 min. Dihydrofuran-3(2H)-one (30 g, 348 mmol) in DMSO (300 mL) was added drop-wise at 0° C. The reaction mixture was stirred at 25° C. for 16 hours. The mixture was poured into ice-water (500 mL) in portions, extracted with DCM (2×500 mL). The combined organic phase was washed with brine (500 mL), dried over Na.sub.2SO4, filtered and concentrated at 30° C. to give 200-DA-C24_8_2 (32 g, crude) as an oil. 3 g from the residue was purified by column (Al.sub.2O.sub.3, PE) to afford 200-DA-C24_8_2 (0.6 g) as an oil.

[0592] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.09-3.90 (m, 4H), 3.03 (d, J=4.4 Hz, 1H), 2.93 (d, J=4.4 Hz, 1H), 2.28 (td, J=8.0, 13.6 Hz, 1H), 1.93 (m, 1H).

Synthesis of ST-200-35-7_1

[0593] ##STR00115##

[0594] A suspension of ST-200-CF3_6C (500 mg, 0.9493 mmol) in THF (2.5 mL) was added dropwise to a solution of n-BuLi (1.13 mL, 2.5 M in hexane, 2.84 mmol) in THF (0.5 mL) at −65° C. under N.sub.2. The mixture was stirred for 30 minutes at −65° C. Diisopropylamine (286 mg, 2.84 mmol) was added at −65° C. Next, 200-DA-C24_8_2 (95.0 mg, 0.9493 mmol) was added drop-wise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 16 hours, quenched by saturated NH.sub.4Cl aqueous (30 mL) and extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-35-7_1 (900 mg, crude) as a solid, which was used directly for the next step.

Synthesis of 1967

[0595] ##STR00116##

[0596] Mg (686 mg, 28.6 mmol) was added to a solution of crude ST-200-35-7_1 (900 mg) in MeOH (10 mL). Next, the reaction mixture was stirred at 60° C. for 2 h under N.sub.2. Aqueous HCl (10 mL, 4 M) was added to the reaction mixture, then 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 in vacuum to give a crude product. The crude product was purified by silica gel chromatography (PE/EtOAc=30/1 to 10/1) to give impure 1967 (460 mg) as a solid. The impure 1967 (460 mg) was purified by re-crystallization from MeCN (2 mL) to give 1967 (175 mg) as a solid. The mother liquid was concentrated in vacuum to give impure ST-200-35-7 (220 mg) as a solid.

[0597] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.10-4.00 (m, 1H), 3.95-3.85 (m, 1H), 3.75-3.65 (m, 1H), 3.55-3.50 (m, 1H), 2.10-2.00 (m, 2H), 2.00-1.85 (m, 3H), 1.85-1.75 (m, 2H), 1.75-1.56 (m, 5H), 1.55-1.40 (m, 6H), 1.40-1.20 (m, 7H), 1.20-1.00 (m, 5H), 1.00-0.88 (m, 4H), 0.85 (s, 3H), 0.75-0.68 (m, 1H), 0.66 (s, 3H).

[0598] LCMS Rt=1.148 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.30H.sub.48F.sub.3NO.sub.3Na [M+MeCN+Na].sup.+ 550, found 550.

Example 20: Synthesis of 2080 and 2081

[0599] ##STR00117##

[0600] Stereochemistry confirmed by Xray data.

[0601] The experimental of Intermediate DA-35-6 can be found in Example 14.

Synthesis of DA-35-4_1A& DA-35-4_1B

[0602] ##STR00118##

[0603] Py (498 mg, 6.30 mmol) and BzCl (531 mg, 3.78 mmol) were added to a solution of DA-35-6 (130 mg, 0.252 mmol) in DCM (5 mL). The mixture was stirred at 25° C. for 6 h and quenched by adding H.sub.2O (5 mL). The mixture was washed with HCl (10 mL, 1 M, aq.), NaHCO.sub.3 (10 mL, sat. aq.), dried over Na.sub.2SO.sub.4, filtered, and concentrated in vacuum to give a crude product. The crude product was purified by silica gel column (PE:EtOAc=20:1 to 10:1) to give DA-35-4_1 (170 mg, impure). The impure DA-35-4_1 (170 mg) was separated by SFC (column: Chiralpak AD-3 50*4.6 mm I.D., 3 um); Condition: Base-IPA; Gradient: 5-40% B; flow rate: 4 mL/min) to give DA-35-41A (56 mg, 36%, Rt=4.889 min, 100% de) and DA-35-4_1B (80 mg, 51%, Rt=5.283 min, 100% de).

[0604] DA-35-4_1A:

[0605] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.04 (d, J=8.0 Hz, 2H), 7.56 (t, J=8.0 Hz, 1H), 7.45 (t, J=8.0 Hz, 2H), 5.04-4.94 (m, 1H), 4.06-3.94 (m, 2H), 3.44-3.32 (m, 2H), 2.10-1.84 (m, 4H), 1.84-1.58 (m, 8H), 1.53-1.23 (m, 12H), 1.22-0.94 (m, 8H), 0.94-0.80 (m, 7H), 0.72-0.57 (m, 4H).

[0606] DA-35-4_1B:

[0607] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.04 (d, J=8.0 Hz, 2H), 7.56 (t, J=8.0 Hz, 1H), 7.45 (t, J=8.0 Hz, 2H), 5.05-4.96 (m, 1H), 4.03-3.93 (m, 2H), 3.44-3.30 (m, 2H), 2.10-1.59 (m, 12H), 1.53-1.23 (m, 12H), 1.22-0.94 (m, 8H), 0.93-0.81 (m, 7H), 0.72-0.60 (m, 4H).

Synthesis of 2080

[0608] ##STR00119##

[0609] A solution of LiOH.H.sub.2O (284 mg, 6.78 mmol) in water (1 mL) was added to a solution of DA-35-4_1A (56 mg, 0.090 mmol) in THF (5 mL) and MeOH (1 mL). The mixture was stirred at 50° C. for 20 h. The mixture was concentrated in vacuum and treated with H.sub.2O (5 mL). The mixture was extracted with EtOAc (3×5 mL). The organic layers were washed with brine (2×15 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated in vacuum. The residue was triturated from MeCN (2 mL) at 25° C. to give 2080 (12 mg, 26%) as a solid.

[0610] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.05-3.95 (m, 2H), 3.40-3.25 (m, 3H), 2.05-1.95 (m, 2H), 1.85-1.80 (m, 2H), 1.75-1.25 (m, 17H), 1.24-0.90 (m, 16H), 0.89-0.75 (m, 3H), 0.65-0.60 (m, 4H).

[0611] LCMS Rt=1.205 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.30H.sub.48F.sub.3O.sub.2 [M+H−H.sub.2O].sup.− 497, found 497.

Synthesis of 2081

[0612] ##STR00120##

[0613] A suspension of LiOH.H.sub.2O (405 mg, 9.67 mmol) in water (1 mL) was added to a solution of DA-35-4_1B (80 mg, 0.129 mmol) in THF (5 mL) and MeOH (1 mL). The mixture was stirred at 50° C. for 20 h. The mixture was concentrated in vacuum and treated with H.sub.2O (5 mL). The mixture was extracted with EtOAc (3×5 mL). The organic layers were washed with brine (2×15 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated in vacuum. The residue was triturated from MeCN (2 mL) at 25° C. to give 2081 (32 mg, 48%) as a solid.

[0614] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.05-3.95 (m, 2H), 3.40-3.25 (m, 3H), 2.05-1.90 (m, 4H), 1.89-1.60 (m, 8H), 1.59-1.35 (m, 10H), 1.34-0.95 (m, 11H), 0.94-0.75 (m, 7H), 0.65-0.60 (m, 4H).

[0615] LCMS Rt=1.205 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.30H.sub.48F.sub.3O.sub.2 [M+H−H.sub.2O].sup.− 497, found 497.

Example 21: Synthesis of 2184

[0616] ##STR00121##

[0617] The synthesis of ST-200-CF3_6C or B7could be found in Example 5.

Synthesis of 200-TBU-E_2

[0618] ##STR00122##

[0619] 200-TBU-E_1 (131 g, 998 mmol) was dissolved in 1690 mL of 5 N hydrochloric acid. The mixture was cooled to 0° C. and a precooled solution of sodium nitrite (109 g, 1.59 mol) in 400 mL of water was added drop-wise, then the reaction mixture was kept below 5° C. After 5 hr, the mixture was stirred at 25° C. for 12 hrs. Solid sodium carbonate (100 g) was added carefully in small portions. The reaction mixture was extracted with isopropyl ether (500 mL*2). The combined organic phases was washed with brine (500 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue was isolated by distillation to afford 200-TBU-E_2 (48 g, 32%) as a solid.

[0620] .sup.1H NMR (400 MHz, CDCl3) δ 4.12 (s, 1H), 1.13 (s, 9H).

Synthesis of 200-TBU-E_2

[0621] ##STR00123##

[0622] LiAlH.sub.4 (14.4 g, 381 mmol) was added to a solution of 200-TBU-E_2 (48 g, 318 mmol) in THF (500 mL) at 0° C. The mixture was warmed to 25° C. and stirred at 25° C. for 30 mins. Water/THF (100 mL, 1/1) was added and the pH was adjusted to 2˜3 with HCl (1 mol/L). The mixture was extracted with EA (2×500 mL), washed with brine (2×200 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated in vacuum to give 200-TBU-E_3 (36 g, crude) as a solid. This product was used in the next step without further purification.

[0623] .sup.1H NMR (400 MHz, CDCl3) δ 3.92-3.86 (m, 2H), 3.68-3.63 (m, 1H), 1.04 (s, 9H).

Synthesis of 200-TBU-E_4

[0624] ##STR00124##

[0625] 200-TBU-E_3 (16 g, 117 mmol) was added to a solution of potassium hydroxide (13.1 g, 234 mmol) in water (13 ml) at 0° C. The ice bath was replaced by a water bath at 20° C. As the cyclization reaction proceeded, a precipitate of potassium chloride formed. After 10 min, the bath temperature was raised slowly to 50° C. The product was isolated by distillation to afford 200-TBU-E_4 (6 g, 51.2%) as an oil. 100% ee after protected with UV group.

[0626] .sup.1H NMR (400 MHz, CDCl3) δ 2.73-2.71 (m, 1H), 2.64-2.63 (m, 1H), 2.62-2.59 (m, 1H), 0.91 (s, 9H).

[0627] Method for ee Checking of Chiral Epoxide

##STR00125##

[0628] n-BuLi (2.5 M, 1.99 mmol, 0.8 mL) was added dropwise to a solution of (methylsulfonyl)benzene (342 mg, 2.19 mmol) in THF (5 mL) was under N.sub.2 at −70° C. After stirring at −70° C. for 30 min, a solution of 200-TBU-E_4 (100 mg, 0.998 mmol) was added. Then reaction was stirred at stirred at 25° C. for 12 hours. The mixture was poured into ice-water (100 mL) and extracted with EA (2×50 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4filtered and concentrated in vacuum. The residue was purified by silica gel chromatography (PE/EA=5/1) to afford 200-TBU-E_4A (80 mg, 31.3%) as an oil. The ee % of product was determined to be 100% by chiral HPLC.

Synthesis of DA-31-2_1

[0629] ##STR00126##

[0630] n-BuLi (0.416 mL, 2.5 M, 1.03 mmol) was added to a solution of diisopropylamine (110 mg, 1.09 mmol) in THF (1 mL) under N.sub.2 at −70° C. The resulting mixture was stirred at 0° C. for 30 min. The mixture was re-cooled to −70° C. To the mixture was added ST-200-CF3_6C (250 mg, 0.474 mmol) in THF (2 mL) at −70° C. The reaction mixture was stirred at −70° C. for 1 hour. (R)-2-(tert-butyl)oxirane (56.8 mg, 0.568 mmol) in THF (1 mL) was added at −70° C. The reaction mixture was warmed to 15° C. slowly and stirred at 15° C. for 16 h. The reaction mixture was quenched with saturated NH.sub.4Cl aqueous (20 mL) at 0° C. The mixture was extracted with EtOAc (2×20 mL). The combined organic phase was washed with brine (10 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give crude DA-31-2_1 (300 mg) as a solid.

[0631] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.95-7.85 (m, 2H), 7.68-7.63 (m, 1H), 7.60-7.50 (m, 2H), 3.45-3.35 (m, 2H), 3.25-3.15 (m, 1H), 2.60-2.55 (m, 1H), 2.10-1.60 (m, 6H), 1.55-1.20 (m, 11H), 1.20-1.00 (m, 7H), 0.93 (s, 9H), 0.90-0.80 (m, 5H), 0.70-0.50 (m, 3H), 0.45 (s, 3H).

Synthesis of DA-31-2

[0632] ##STR00127##

[0633] Mg (229 mg, 9.55 mmol) was added to a solution of DA-31-2_1 (300 mg, 0.478 mmol) in MeOH (5 mL). Next, the reaction was stirred at 60° C. for 2 h under N.sub.2. Aqueous HCl (10 mL, 4 M) was added to the reaction mixture, then 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 in vacuum to give a crude product. The crude product was purified by silica gel chromatography (PE/EtOAc=30/1 to 10/1) to give impure DA-31-2 (100 mg, impure) as a solid. Dry Pd(OH).sub.2/C (50 mg) was added to a solution of DA-31-2 (100 mg, impure, 0.205 mol) in MeOH/THF=1/1 (4 mL). Next, the reaction mixture was stirred at 50° C. for 16 h under H.sub.2 and 50 Psi. The reaction mixture was filtered through a pad of Celite and washed with THF (3×5 mL). The combined organic layer was concentrated in vacuum to give a crude DA-31-2 (85 mg) as a solid, which was purified by re-crystallization from MeCN (2 mL) to give DA-31-2 (60 mg, 71%) as a solid.

[0634] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.20-3.05 (m, 1H), 2.10-1.90 (m, 3H), 1.90-1.60 (m, 7H), 1.55-1.40 (m, 5H), 1.40-1.10 (m, 14H), 1.10-1.00 (m, 3H), 0.93 (s, 9H), 0.89 (s, 3H), 0.75-0.66 (m, 1H), 0.65 (s, 3H).

[0635] LCMS Rt=1.356 min in 2.0 min chromatography, 30-90 AB, purity 99%, MS ESI calcd. for C.sub.29H.sub.48F.sub.3O [M−H.sub.2O+H].sup.+ 469, found 469.

Example 22: Synthesis of 2285

[0636] ##STR00128##

[0637] The synthesis of ST-200-CF3_6C or B7 can be found in Example 5.

Synthesis of ST-200-3CF3-A7R_1

[0638] ##STR00129##

[0639] A suspension of ST-200-CF36C (250 mg, 0.475 mmol) in THF (2.5 mL) was added dropwise to a solution of n-BuLi (568 μL, 2.5 M in hexane, 1.42 mmol) in THF (0.5 mL) at −78° C. under N.sub.2. The mixture was stirred for 30 minutes at −78° C. A solution of 2-(methyl)oxirane (41.3 mg, 0.712 mmol) was added dropwise at −78° C. The mixture was stirred for another 30 min and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 16 hour. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL), extracted with EtOAc (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-3CF3-A7R_1 (340 mg, crude) as a solid, which was used directly for the next step.

Synthesis of 2285

[0640] ##STR00130##

[0641] Mg powder (556 mg, 23.2 mmol) was added to a solution of ST-200-3CF3-A7R_1 (340 mg, 0.581 mmol) in dry methanol (30 mL) under N.sub.2 at 60° C. The reaction mixture was quenched by 2 M HCl (50 mL) added dropwise at 10° C. until the solid was dissolved. After extraction with EtOAc (2×50 mL), the organic layer was washed with sat. NaHCO.sub.3 (50 mL), brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column, eluted with PE/EtOAc=20/l to 5/1, to give 2285 (80 mg, impure containing some 22-23 olefin) as a solid, which was used for next step without further purification.

Synthesis of ST-200-3CF3-A7R

[0642] ##STR00131##

[0643] Pd(OH).sub.2 (20%, 126 mg, 0.180 mmol) was added to a solution of 2285 (80 mg, 0.180 mmol) in MeOH/THF (10 mL/10 mL) under Ar. After degassing three times with N.sub.2 and H.sub.2, the reaction mixture was stirred for 16 h at 50° C. under H.sub.2 atmosphere (50 psi). The desired product was produced, the catalyst was removed by suction, and the filtrate was concentrated to give 2285 (50 mg, impure) as a solid, which was triturated with MeCN (3 mL) at 25° C. to give 2285 (36 mg, 45%) as a solid.

[0644] 2285

[0645] .sup.1HNMR (400 MHz, CDCl3) δ 3.74-3.72 (m, 1H), 2.08-2.06 (m, 1H), 2.00-1.91 (m, 2H), 1.88-1.75 (m, 2H), 1.74-1.59 (m, 3H), 1.52-1.22 (m, 13H), 1.21-0.96 (m, 10H), 0.95-0.86 (m, 4H), 0.85 (s, 3H), 0.73-0.62 (m, 4H)

[0646] LCMS Rt=1.199 min in 2 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.26H.sub.42F.sub.3O [M+H−H.sub.2O]+ 427, found 427.

Example 23: Synthesis of 2392

[0647] ##STR00132##

[0648] The experimental of intermediate ST-200-CF3_4A or A7 can be found in Example 3.

Synthesis of ST-200-3CF3-C14_1

[0649] ##STR00133##

[0650] BuLi (0.476 mL, 2.5 M in hexane, 1.19 mmol) was added to THF (0.5 mL). A solution of ST-200-CF3_4A (250 mg, 0.476 mmol) in THF (3 mL) was added at −70° C. The mixture was stirred at −70° C. for 1 h. 6,6-difluoro-1-oxaspiro[2.5]octane (210 mg, 1.42 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 h. The mixture was warmed to 25° C. and stirred for 16 hrs. NH.sub.4Cl (50 mL, sat. aq.) was added to the mixture, then the mixture was extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give ST-200-3CF3-C14_1 (300 mg, crude) as a solid, which was used directly for the next step.

Synthesis of 2392

[0651] ##STR00134##

[0652] A solution of ST-200-31-15_1 (300 mg, 0.445 mmol) in MeOH (20 mL) was heated at 55° C. Mg powder (427 mg, 17.8 mmol) was added in one portion at 55° C. The mixture was refluxed at 65° C. for 1 h. The mixture was quenched with HCl (50 mL, 1N) until the reaction became clear, then was extracted with DCM (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give impure product (110 mg), which was purified again by SFC (column: AD (250 mm*30 mm, 5 um), gradient: 35-35% B (A=0.1% NH3/H2O, B=MeOH), flow rate: 60 mL/min) to give 2392 (72 mg, 30%) as a solid.

[0653] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.38-5.35 (m, 1H), 2.49 (s, 2H), 2.20-1.81 (m, 9H), 1.80-1.71 (m, 3H), 1.70-1.58 (m, 5H), 1.56-1.36 (m, 7H), 1.35-1.22 (m, 2H), 1.20-1.08 (m, 4H), 1.06 (s, 3H), 1.04-0.92 (m, 6H), 0.68 (s, 3H).

[0654] LCMS Rt=1.248 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.30H.sub.44F.sub.5O [M+H−H.sub.2O].sup.+ 515, found 515.

Example 24: Synthesis of 2499

[0655] ##STR00135##

[0656] The synthesis of DA-31-10_2 can be found in Example 11.

Synthesis of 2499

[0657] ##STR00136##

[0658] To a suspension of LiAlH.sub.4 (1.03 g, 27.4 mmol) in THF (80 mL) was added a solution of DA-31-10_2 (6.3 g, 13.7 mmol) in THF (20 mL) under N.sub.2 dropwise at 0° C. The reaction was stirred at 25° C. for 2 h. The reaction was quenched with water/THF (1/10, 40 mL). To the mixture was added 2 M HCl (100 mL) at 0° C. and extracted with EtOAc (2×100 mL). The combined organic phase was washed with brine (300 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to afford 2499 (5 g, crude) as a solid. 100 mg of the impure DA-31-10_3 was triturated with CH.sub.3CN (5 mL) at 25° C. for 3 hours to give 2499 (52 mg, 52%) as a solid.

[0659] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.70-3.50 (m, 2H), 2.10-1.90 (m, 3H), 1.85-1.75 (m, 2H), 1.70-1.60 (m, 4H), 1.50-1.20 (m, 14H), 1.15-0.80 (m, 12H), 0.70-0.60 (m, 4H).

[0660] LCMS Rt=1.179 min in 2 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. For C.sub.25H.sub.40F.sub.3O [M+H−H.sub.2O]+ 413, found 413.

Example 25: Synthesis of 2500

[0661] ##STR00137##

[0662] The experimental of intermediate ST-200-CF3_4A can be found in Example 3.

Synthesis of ST-200-31-6_1

[0663] ##STR00138##

[0664] n-BuLi (568 μL, 2.5 M in hexane, 1.42 mmol) was added to a solution of diisopropylamine (143 mg, 1.42 mmol) in THF (0.5 mL) at −78° C. under N.sub.2. A suspension of ST-200-CF3_4A (250 mg, 0.476 mmol) in THF (2.5 mL) was added dropwise. The mixture was stirred for 30 minutes at −78° C. A solution of 2-(tert-butyl)oxirane (71.5 mg, 0.715 mmol) was added dropwise at −78° C. The mixture was stirred for another 30 min and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 16 hour. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL), extracted with EtOAc (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-31-6_1 (350 mg, crude) as a solid, which was used directly for the next step.

Synthesis of 2500

[0665] ##STR00139##

[0666] A solution of ST-200-31-6_1 (350 mg, 0.6081 mmol) in MeOH (25 mL) was heated at 60° C. Mg powder (584 mg, 24.3 mmol) was added in four portions at 60° C. The mixture was stirred at 60° C. for 1 h. The mixture was quenched with HCl (50 mL, 2 M) until the reaction became clear and extracted with DCM (2×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give 112 mg of impure product as a solid, which was triturated with MeCN (3 mL) at 25° C. to give 70 mg as a solid. The 70 mg product was dissolved in THF (8 mL) and treated with Lindlar (100 mg) under N.sub.2. The mixture was degassed under vacuum and purged with H.sub.2(15 psi) several times. The mixture was stirred for 2 hrs at 25° C. under H.sub.2(15 psi). The mixture was filtered and the filter was concentrated in vacuum. The residue was purified by flash column (0-20% EtOAc in PE) to afford pure 2500 (20 mg) as a solid

[0667] .sup.1H NMR (CDCl3, 400 MHz) δ 5.40-5.30 (m, 1H), 3.20-3.00 (m, 1H), 2.50-2.45 (s, 2H), 2.05-2.00 (m, 4H), 1.96-1.33 (m, 13H), 1.33-1.20 (m, 7H), 1.20-0.80 (m, 16H), 0.68 (s, 3H).

[0668] LCMS Rt=1.404 min in 2 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.28H.sub.46F.sub.3O [M−H.sub.2O+H].sup.+ 467, found 467.

Example 26: Synthesis of 2602

[0669] ##STR00140##

[0670] The experimental of intermediate ST-200-CF3_4A or A7 can be found in Example 3.

Synthesis of ST-200-3CF3_C7S_1

[0671] ##STR00141##

[0672] A suspension of ST-200-CF34A (250 mg, 0.476 mmol) in THF (2.5 mL) was added dropwise to a solution of n-BuLi (0.568 mL, 2.5 M in hexane, 1.42 mmol) in THF (0.5 mL) at −65° C. under N.sub.2. The mixture was added diisopropylamine (143 mg, 1.42 mmol) and stirred for 30 minutes at −65° C. A solution of (S)-2-methyloxirane (33.1 mg, 0.571 mmol) was added dropwise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 16 hours. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL) and extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-3CF3-C7S_1 (250 mg, crude) as a solid, which is used directly for the next step.

Synthesis of 2602

[0673] ##STR00142##

[0674] Mg powder (415 mg, 17.1 mmol) was added to a solution of ST-200-3CF3-C7S_1 (250 mg, 0.428 mmol) and nickel (II) chloride (13.8 mg, 0.107 mmol) in dry methanol (20 mL) under N.sub.2 and the mixture was stirred at 50° C. to initiate continuous hydrogen generation. The reaction mixture was stirred at 60° C. for 1 hour. Next, the reaction mixture was quenched by 2M HCl (100 mL) which was added dropwise at 10° C. until solid was dissolved. After extracting with EtOAc (2×150 mL), the combined organic layer was washed with sat. NaHCO.sub.3 aq. (300 mL), brine (300 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a solid, which was purified by silica gel chromatography (PE:EtOAc=4:1) to give 100 mg of solid (the residue was containing 13% 22, 23 alkene). The impure residue was dissolved in THF (20 mL) was added Lindlar (15.9 mg, 0.225 mmol) under N.sub.2. The mixture was degassed under vacuum and purged with H.sub.2 several times. The mixture was stirred for 2 hrs at 25° C. under H.sub.2. The mixture was filtered and the filter was concentrated in vacuum. The residue was purified by SFC (column: C2 250 mm*30 mm, 10 um), gradient: 35-35% B (A=0.1% NH3/H2O, B=EtOH), flow rate: 50 mL/min) to give 2602 (16 mg, 54%) as a solid.

[0675] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-35 (m, 1H), 3.75-3.65 (m, 1H), 2.50-2.45 (m, 2H), 2.10-1.70 (m, 7H), 1.69-1.50 (m, 6H), 1.49-1.20 (m, 10H), 1.19-0.90 (m, 11H), 0.68 (s, 3H).

[0676] LCMS Rt=1.202 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.26H.sub.40F.sub.3O [M+H−H.sub.2O].sup.− 425, found 425.

Example 27: Synthesis of 2706 and 2707

[0677] ##STR00143##

[0678] The experimental of intermediate ST-200-CF3_4A can be found in Example 3. ST-200-35-7 can be found in Example 19. The stereochemistry of 2707 was confirmed by X-ray.

Synthesis of ST-200-35-8A/8B

[0679] ##STR00144##

[0680] BzCl (258 mg, 1.84 mmol) was added to a solution of ST-200-35-7 (300 mg, 0.616 mmol) in pyridine (5 mL) at 0° C. The mixture was stirred for 1 h at 0° C. To the mixture was added water (10 mL) at 0° C. and extracted with DCM (3×10 mL). The organic layer was washed with 1M HCl (10 mL), saturated Na.sub.2CO.sub.3 (10 mL) and brine. The mixture was dried over anhydrous Na.sub.2SO.sub.4, concentrated in vacuum to give a residue. The residue was purified by prep-TLC (PE/EA=5/1) to give a mixture. The mixture was separated by SFC twice (Instrument: MG-II; Method: Column: AD (250 mm*30 mm, 5 um); Condition: 0.1% NH3H2O ETOH; Begin B: 40%; End B: 40%; FlowRate (ml/min): 60; Injections: 90) to give peak 1 (Rt=5.134 min) as ST-200-35-8B (44 mg, 12%) and peak 2 (Rt=5.766 min) ST-200-35-8A (38 mg, 10%) both as a solid.

[0681] ST-200-35-8B:

[0682] SFC Rt=5.134 min in 10.0 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100% de.

[0683] ST-200-35-8A:

Synthesis of 2706

[0684] ##STR00145##

[0685] MeOH (0.2 mL), water (0.2 mL) and LiOH.H.sub.2O (31.2 mg, 0.744 mmol) were added to a solution of ST-200-35-8B (44 mg, 0.0744 mmol) in THF (0.4 mL). The mixture was stirred at 50° C. for 16 h. EtOAc (5 mL) and water (2 mL) were added to the mixture. The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum and triturated from MeCN (1 mL) to give 2706 (24 mg, 66%) as a solid.

[0686] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.02 (q, J=8.0 Hz, 1H), 3.93-3.83 (m, 1H), 3.69 (d, J=9.2 Hz, 1H), 3.55 (d, J=9.2 Hz, 1H), 2.10-1.79 (m, 7H), 1.75-1.59 (m, 5H), 1.55-0.99 (m, 18H), 0.98-0.88 (m, 4H), 0.85 (s, 3H), 0.75-0.60 (m, 4H).

[0687] HPLC Rt=3.97 min in 8.0 min chromatography, 50-100_AB_E, purity 100%.

[0688] MS ESI calcd. for C.sub.28H.sub.44F.sub.3O.sub.2 [M+H−H.sub.2O].sup.+ 469.3288, found 469.3244.

Synthesis of 2707

[0689] ##STR00146##

[0690] MeOH (0.2 mL), water (0.2 mL) and LiOH.H.sub.2O (26.9 mg, 0.642 mmol) were added to a solution of ST-200-35-8A (38 mg, 0.0643 mmol) in THF (0.4 mL). The mixture was stirred at 50° C. for 16 h. EtOAc (5 mL) and water (2 mL) were added to the mixture. The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum and triturated from MeCN (1 mL) to give 2707 (21 mg, 67%) as a solid.

[0691] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.02 (q, J=8.0 Hz, 1H), 3.94-3.85 (m, 1H), 3.69 (d, J=9.2 Hz, 1H), 3.54 (d, J=9.2 Hz, 1H), 2.10-1.59 (m, 13H), 1.55-0.99 (m, 17H), 0.98-0.88 (m, 4H), 0.85 (s, 3H), 0.75-0.62 (m, 4H).

[0692] HPLC Rt=3.93 min in 8.0 min chromatography, 50-100_AB_E, purity 100%.

[0693] MS ESI calcd. for C.sub.28H.sub.44F.sub.3O.sub.2 [M+H−H.sub.2O].sup.+ 469.3288, found 469.3244.

Example 28: Synthesis of E-2817

[0694] ##STR00147##

[0695] The synthesis of ST-200-CF3_6C can be found in Example 5.

The Synthesis of ST-200-43-4_2

[0696] ##STR00148##

[0697] To a suspension of t-BuOK (3.53 g, 31.6 mmol) in THF (30 mL) was added Me.sub.3SI (4.18 g, 20.5 mmol) under N.sub.2 at 15° C. The suspension was stirred at 15° C. for 30 min. To the mixture was added a solution of 200-DA-E31_1A (2 g, 15.8 mmol) in 10 ml of THF dropwise at 15° C. The mixture was stirred at 15° C. for 16 hrs. The mixture was quenched with sat. NH.sub.4Cl (100 mL) and extracted with EtOAc (3×150 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, and concentrated in vacuum to give 200-DA-E31_1 (1.8 g, 81%) as a liquid.

[0698] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.58 (s, 2H), 1.90-1.80 (m, 1H), 1.70-1.55 (m, 2H), 1.54-1.45 (m, 3H), 1.40-1.30 (m, 2H), 1.00-0.90 (m, 6H).

Synthesis of ST-200-3CF3-A18_1

[0699] ##STR00149##

[0700] First, n-BuLi (0.5 mL, 2.5 M in hexane, 1.25 mmol) was added To THF (0.5 mL). A solution of ST-200-CF3_6C (250 mg, 0.4746 mmol) in THF (3 mL) was added at −70° C. The mixture was stirred at −70° C. for 1 h. ST-200-43-4_2 (133 mg, 0.9492 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 h. The mixture was warmed to 25° C. and stirred for 16 hrs. The reaction mixture was quenched by adding NH.sub.4Cl (50 mL, sat. aq.) and extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give ST-200-3CF3-A18_1 (390 mg, crude) a solid, which was used directly for the next step.

Synthesis of E-2817

[0701] ##STR00150##

[0702] A solution of ST-200-3CF3-A18_1 (390 mg, 0.5847 mmol) in MeOH (25 mL) was heated at 60° C. Mg powder (500 mg, 20.8 mmol) was added in four portions at 60° C. The mixture was stirred at 60° C. for 1 h. The mixture was quenched with HCl (50 mL, 2 M) until the reaction became clear and extracted with DCM (2×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give 135 mg of a solid. The impure product was purified by flash column (0-20% of EtOAc in PE) to give E-2817 (101 mg, 75%) as a solid.

[0703] .sup.1H NMR (CDCl3, 400 MHz) δ 2.08-2.03 (m, 1H), 1.98-1.88 (m, 2H), 1.78-1.73 (m, 2H), 1.73-1.60 (m, 3H), 1.60-1.45 (m, 12H), 1.45-1.27 (m, 7H), 1.27-1.19 (m, 9H), 1.19-1.00 (m, 6H), 0.93-0.84 (m, 9H), 0.75-0.64 (s, 4H).

[0704] LCMS Rt=1.463 min in 2 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.32H.sub.52F.sub.3O [M+H−H.sub.2O].sup.+ 509, found 509.

Example 29: Synthesis of 2918

[0705] ##STR00151##

[0706] The synthesis of ST-200-CF3_6C can be found in Example 5.

Synthesis of ST-200-3CF3-A8_1

[0707] ##STR00152##

[0708] A suspension of ST-200-CF3_6C (250 mg, 0.475 mmol) in THF (2.5 mL) was added dropwise to a solution of n-BuLi (568 μL, 2.5 M in hexane, 1.42 mmol) in THF (0.5 mL) at −78° C. under N.sub.2. The mixture was stirred for 30 min at −78° C. A solution of 2-(trifluoromethyl)oxirane (79.7 mg, 0.712 mmol) was added dropwise at −78° C. The mixture was stirred for another 30 min and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 16 hours. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL) and extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-3CF3-A8_1 (340 mg, crude) as a solid, which was used directly for the next step.

Synthesis of 2918

[0709] ##STR00153##

[0710] A solution of ST-200-3CF3-A8_1 (340 mg, 0.5322 mmol) in MeOH (25 mL) was heated at 60° C. Mg powder (508 mg, 21.2 mmol) was added in four portions at 60° C. The mixture was stirred at 60° C. for 1 h. The mixture was quenched with HCl (50 mL, 1N) until the reaction became clear and extracted with DCM (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give 63 mg of a solid, which was triturated from DCM and hexane to give 2918 (5 mg, 2%).

[0711] .sup.1H NMR (CDCl3, 400 MHz) δ 3.90-3.80 (m, 1H), 2.20-1.70 (m, 6H), 1.70-1.50 (m, 7H), 1.50-1.25 (m, 5H), 1.25-1.10 (m, 5H), 1.10-0.80 (m, 12H), 0.70-0.65 (m, 4H).

[0712] LCMS Rt=1.219 min in 2 min chromatography, 30-90 AB, purity 100%.

Example 30: Synthesis of 3035

[0713] ##STR00154##

[0714] The experimental of intermediate ST-200-CF3_4A can be found in Example 3.

The Synthesis of the Tosylate

[0715] ##STR00155##

[0716] To a suspension of LiAlH.sub.4 (45.3 g, 1.26 mol) in THF (1 L) was added dropwise a solution of 7330_3S (100 g, 632 mmol) in THF (500 mL) at 0° C. and the inner temperature raised to about 50° C. After addition, the mixture was stirred at 70° C. for 16 hours. The mixture was quenched with HCl (1 L, 3 M aq.) to pH=2 and extracted with MTBE (3×500 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure (<40° C.) to give 7330_4S (92 g, crude) as an oil.

[0717] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.96-3.92 (m, 1H), 3.58-3.53 (m, 1H), 3.08 (s, 1H), 1.98-1.89 (m, 1H), 1.38 (s, 3H).

[0718] To a solution of 7330_4S (50 g, 346 mmol) in pyridine (300 mL) was added 4-methylbenzene-1-sulfonyl chloride (98.9 g, 519 mmol) in portions during 5 minutes at 0° C. The reaction solution was stirred at 20° C. for 16 hrs. The reaction mixture was quenched with 2N HCl (400 mL) to pH=1-2 at 0° C. The inner temperature was maintained below 30° C. and the mixture was extracted with MTBE (3×200 mL). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by column (0-10% of EtOAc in PE) to give 7330_5S (93 g, 90%, 99.42% ee) as an oil.

[0719] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.79 (d, J=7.6 Hz, 2H), 7.37 (d, J=8.0 Hz, 2H), 4.13-4.03 (m, 2H), 2.99 (s, 1H), 2.46 (s, 3H), 1.37 (s, 3H),

[0720] LCMS Rt=1.103 min in 2.0 min chromatography, 10-80 AB, purity 100%, no MS detected.

Synthesis of ST-200-3CF3-C11S_1

[0721] ##STR00156##

[0722] A suspension of ST-200-CF34A (250 mg, 0.48 mmol) in THF (4 mL) was added dropwise to a solution of n-BuLi (0.48 mL, 2.5 M in hexane, 1.19 mmol) in THF (1 mL) at −70° C. under N.sub.2. After stirring for 30 minutes at −70° C., diisopropylamine (120 mg, 1.19 mmol) was added dropwise at −70° C., followed by adding (S)-3,3,3-trifluoro-2-hydroxy-2-methylpropyl 4-methylbenzenesulfonate (212 mg, 0.71 mmol) dropwise at −70° C. The mixture was stirred for another 30 min and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 24 hour. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (5 mL), extracted with EtOAc (3×10 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-3CF3-C11S_1 (480 mg, crude), which was used directly.

Synthesis of 3035

[0723] ##STR00157##

[0724] Mg powder (705 mg, 29.4 mmol) and NiCl.sub.2 (1 mg, 0.007 mmol) were added with stirring to a solution of ST-200-3CF3-C11S_1 (480 mg, 0.74 mmol) in 50 mL of anhydrous MeOH under N.sub.2 at 60° C. The reaction mixture was quenched by 2 M HCl (10 mL) until the solid was dissolved. The mixture was extracted with EtOAc (3×20 mL). The combined organic layer was washed with sat. NaHCO.sub.3 (50 mL), brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-20% of EtOAc in PE) to give a crude product, which was further purified by re-crystallized from MeCN (10 mL) at 85° C. to give 3035 (53 mg, 21%) as a solid.

[0725] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.41-5.34 (m, 1H), 2.53-2.46 (s, 2H), 2.08-1.92 (m, 4H), 1.91-1.58 (m, 7H), 1.54-1.35 (m, 7H), 1.33-1.30 (s, 3H), 1.29-1.08 (m, 5H), 1.07-1.05 (s, 3H), 1.05-0.91 (m, 5), 0.73-0.63 (s, 3).

[0726] LCMS Rt=1.213 min in 2 min chromatography, 30-90AB_2MIN_E, purity 99%.

Example 31: Synthesis of 3149

[0727] ##STR00158##

[0728] The experimental of intermediate ST-200-CF3_4A can be found in Example 3.

Synthesis of ST-200-3CF3_C8R_1

[0729] ##STR00159##

[0730] A suspension of ST-200-CF34A (250 mg, 0.476 mmol) in THF (2.5 mL) was added dropwise to a solution of n-BuLi (0.568 mL, 2.5 M in hexane, 1.42 mmol) in THF (0.5 mL) at −65° C. under N.sub.2. After adding diisopropylamine (143 mg, 1.42 mmol) and stirring for 30 minutes at −65° C., a solution of (R)-2-(trifluoromethyl) oxirane (63.9 mg, 0.571 mmol) was added dropwise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 16 hours. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL) and extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-3CF3-C8R_1 (250 mg, crude) as a solid, which was used directly for the next step.

Synthesis of 3149

[0731] ##STR00160##

[0732] Mg powder (379 mg, 15.6 mmol) was added to a solution of ST-200-3CF3-C8R_1 (250 mg, 0.392 mmol) and nickel (II) chloride (12.7 mg, 0.098 mmol) in dry methanol (50 mL) under N.sub.2 at 50° C. While adding Mg, the mixture was stirred to initiate continuous hydrogen generation. Next, the reaction mixture was stirred at 60° C. for 1 hour. The reaction mixture was quenched by 2M HCl (100 mL) which was added dropwise at 10° C. until solid was dissolved. After extracting with EtOAc (2×150 mL), the combined organic layer was washed with sat. NaHCO.sub.3 aq. (300 mL), brine (300 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a solid, which was purified by silica gel chromatography (PE/THF=4/1) to give a crude product, which was re-crystallized from MeCN (10 mL) to give a impure product(30 mg, 15%). The impure product (30 mg, 0.068 mmol) was purified by SFC (column: AD 250 mm*30 mm, 10 um), gradient: 20-20% B (A=0.1% NH.sub.3/H2O, B=EtOH), flow rate: 60 mL/min) to give 3149 (12 mg, 40%) as a solid.

[0733] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 3.75-3.65 (m, 1H), 2.50-2.45 (m, 2H), 2.10-1.70 (m, 11H), 1.69-1.50 (m, 10H), 1.49-0.90 (m, 10H), 0.69 (s, 3H).

[0734] HPLC Rt=6.25 min in 1.2 min chromatography, 30-90 AB, purity 98%.

[0735] HRMS ESI calcd. for C.sub.26H.sub.39F.sub.6O.sub.2 [M+H].sup.− 497.2849, found 497.2842.

Example 32: Synthesis of 3266

[0736] ##STR00161##

[0737] The experimental of intermediate ST-200-CF3_4A can be found in Example 3.

Synthesis of ST-200-3CF3-C7R_1

[0738] ##STR00162##

[0739] A suspension of ST-200-CF3_4A (250 mg, 0.476 mmol) in THF (4 mL) was added dropwise to a solution of n-BuLi (568 mL, 2.5 M in hexane, 1.42 mmol) in THF (1 mL) at −65° C. under N.sub.2. After stirring at −65° C. 30 minutes, diisopropylamine (143 mg, 1.42 mmol) was added at −65° C. After that, (R)-2-methyloxirane (82.4 mg, 1.42 mmol) was added dropwise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 16 hours. The reaction was quenched with sat. NH.sub.4Cl aq. (50 mL), extracted with EtOAc (3×50 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated to give a crude product as a solid, which was used directly for the next step.

Synthesis of 3266

[0740] ##STR00163##

[0741] Mg powder (410 mg, 17.1 mmol) was added in four portions by stirring into a solution of ST-200-3CF3_C7R_1 (250 mg, 0.428 mmol) and NiCl.sub.2 (5.52 mg, 0.043 mmol) in dry methanol (20 mL) under N.sub.2 at 50° C. After stirring at 60° C. for 1 hour, the mixture was quenched with HCl (50 mL, 1N) until the reaction became clear and extracted with EtOAc (3×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated. The residue was purified by flash column (0-15% of EtOAc in PE) to give an impure product (100 mg, 0.225 mmol, impure, containing 13% 22, 23 alkene). Lindlar catalyst (200 mg, 0.225 mmol) was added to a solution of impure product in THF (20 mL) under N.sub.2. The mixture was degassed under vacuum and purged with H.sub.2 several times. The mixture was stirred for 2 hours at 25° C. The reaction mixture was filtered through a pad of Celite and washed with THF (3×10 mL). The filtrate was concentrated to give a impure product, which was triturated from n-hexane(10 mL) at 68° C. for 2 hours to give a impure product as a solid. The impure product was purified by silica gel chromatography (PE/EtOAc=0 to 5/1) to give 3266 (48 mg, impure) as a solid, which was purified by SFC(Column: AD (150×4.6 mm, 3 um), Gradient: 5%-40% B (A: CO.sub.2 B: ethanol) Flow rate: 2.5 mL/min) to afford 3266 (10 mg) as a solid.

[0742] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.33. (m, 1H), 3.78-3.65 (m, 1H), 2.52-2.45 (m, 2H), 2.08-1.65 (m, 7H), 1.58-1.32 (m, 7H), 1.32-1.23 (m, 4H), 1.23-0.75 (m, 16H), 0.68 (s, 3H).

[0743] LCMS Rt=1.149 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.26H.sub.40F.sub.3O [M+H−H.sub.2O].sup.+ 425, found 425.

Example 33: Synthesis of 3382

[0744] ##STR00164##

[0745] Stereochemistry was assigned based on synthesis with chiral epoxide, see Example 35 for synthesis.

[0746] The experimental of intermediate ST-200-CF3_4A can be found in Example 3.

Synthesis of ST-200-31-6_1

[0747] ##STR00165##

[0748] A suspension of ST-200-CF3-4A (500 mg, 0.95 mmol) in THF (4 mL) was added dropwise to a solution of n-BuLi (0.95 mL, 2.5 M in hexane, 2.38 mmol) in THF (1 mL) at −70° C. under N.sub.2. After stirring for 30 minutes at −70° C., a solution of diisopropylamine (240 mg, 2.38 mmol) was added dropwise at −70° C., followed by adding a solution of 2-(tert-butyl)oxirane (142 mg, 1.42 mmol) dropwise at −70° C. The mixture was stirred at −70° C. for another 30 min and then warmed to 25° C. gradually. After stirring for at 25° C. for 24 hour, the reaction mixture was quenched by saturated NH.sub.4Cl aqueous (5 mL), extracted with EtOAc (3×20 mL). The combined organic phase was washed with brine (40 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-31-6_1 (650 mg, crude), which was used directly.

Synthesis of ST-200-31-6

[0749] ##STR00166##

[0750] Mg powder (998 mg, 41.6 mmol) and NiCl.sub.2 (5 mg, 0.05 mmol) were added with stirring to a solution of ST-200-31-6 (650 mg, 1.04 mmol) in 100 mL of anhydrous MeOH under N.sub.2 at 60° C. The reaction mixture was quenched by 2 M HCl (50 mL) until solid was dissolved. The mixture was extracted with EtOAc (3×100 mL). The combined organic layer was washed with sat. NaHCO.sub.3 (150 mL), brine (150 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-15% of EtOAc in PE) to give impure ST-200-31-6 as a solid. Lindlar catalyst (200 mg) was added to a solution of the ST-200-31-6 in EtOAc (10 mL) under N.sub.2. The suspension was degassed under vacuum and purged with H.sub.2 for three times. Then the solution was hydrogenated under 15 psi of hydrogen at 25° C. for 4 h. The mixture was filtered through a pad of celite and washed with EtOAc (3×10 mL). The filtrate was concentrated and concentrated to give ST-200-31-6 (210 mg, 43%) as a solid.

[0751] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.39-5.34 (m, 1H), 3.18-3.06 (m, 1H), 2.49 (s, 2H), 2.17 (s, 1H), 2.02-1.58 (m, 7H), 1.53-1.29 (m, 9H), 1.22-0.97 (m, 10H), 0.95-0.84 (m, 13H), 0.72-0.65 (m, 3H).

Synthesis of 3382

[0752] ##STR00167##

[0753] ST-200-31-6 (210 mg, 0.43 mmol) was purified by SFC (column: AD (250 mm*30 mm, 10 um)), gradient: 20-20% B (A=0.1% NH.sub.3/H.sub.2O, B=EtOH), flow rate: 50 mL/min) to give 3382 (90 mg, 43%) as a solid.

[0754] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.34 (m, 1H), 3.19-3.12 (m, 1H), 2.48 (s, 2H), 2.09-1.67 (m, 8H), 1.53-1.23 (m, 12H), 1.22-0.98 (m, 8H), 0.95-0.84 (m, 12H), 0.69 (s, 3H).

[0755] LCMS Rt=1.440 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.29H.sub.46F.sub.3O [M+H−H.sub.2O].sup.+ 467, found 467.

[0756] SFC_E1 Rt=4.337 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, purity: 100%.

Example 34: Synthesis of 3495 and 3496

[0757] ##STR00168##

[0758] The stereochemistry for 3496 was determined by X-ray data. The experimental of intermediate ST-200-CF3_4A can be found in Example 3.

Synthesis of 200-DA-C24_8_2

[0759] ##STR00169##

[0760] Sodium hydride (5.98 g, 60% in mineral oil, 150 mmol) was added in portions to a mixture of trimethylsulfonium iodide (30.6 g, 150 mmol) in THF (100 mL) at 0° C. under N.sub.2. The mixture was stirred at 0° C. for 30 min. Dihydrofuran-3(2H)-one (10 g, 116 mmol) in DMSO (100 mL) was added dropwise at 0° C. The reaction mixture was stirred at 0° C. for 2 hours. The mixture was poured in portions into ice-water (500 mL) and extracted with DCM (2×500 mL). The combined organic phase was washed with brine (500 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to afford 200-DA-C24_8_2 (4 g, crude, 34%) as an oil at 18° C., which was used directly for the next step.

Synthesis of ST-200-CF3_8

[0761] ##STR00170##

[0762] Butyllithium (2.71 mL, 2.5 M in n-hexane, 6.79 mmol) was added to a solution of diisopropylamine (714 mg, 7.33 mmol) in THF (3 mL) at −70° C. The mixture was warmed to 0° C. and stirred at 0° C. for 30 minutes. The mixture was cooled to −70° C. and 200-DA-C24_8_2 (300 mg, 2.99 mmol) in THF (2 mL) was added. The mixture was stirred at −70° C. for 1 h. ST-200-CF3_4A (1.42 g, 2.71 mmol) in THF (2 mL) was added at −70° C. The mixture was warmed to 25° C. and stirred at this temperature for 16 hours. The mixture was quenched with Sat NH.sub.4Cl (10 mL). The mixture was extracted with EtOAc (2×10 mL). The organic phase was washed with brine (2×10 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum. The crude product purified by flash column (0-50% of EtOAc in PE) to give ST-200-CF3_8 (280 mg, 17%) as a solid, which was used directly for the next step.

Synthesis of Compound 10

[0763] ##STR00171##

[0764] Nickel (II) chloride (580 μg, 4.48 μmol) and Mg powder (435 mg, 17.9 mmol) were added in four portions to a solution of ST-200-CF3_8 (280 mg, 0.448 mmol) in 50 mL of dry methanol under N.sub.2 at 60° C. The reaction mixture was quenched by 1M HCl (150 mL) which was added dropwise until solid was dissolved. After extracting with EtOAc (3×50 mL), the organic layer was washed with sat. NaHCO.sub.3 (50 mL), brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-20% of EtOAc in PE) to give Compound 10 (210 mg, 97%) as a solid.

[0765] .sup.1H NMR CDCl.sub.3 400 MHz δ 5.39-5.35 (m, 1H), 3.93-3.82 (m, 1H), 3.72-3.68 (m, 1H), 3.59-3.51 (m, 1H), 2.49 (s, 2H), 2.10-1.80 (m, 8H), 1.80-1.62 (m, 4H), 1.60-1.39 (m, 7H), 1.39-1.12 (m, 6H), 1.12-0.91 (m, 9H), 0.69 (s, 3H).

Synthesis of 3495 and 3496

[0766] ##STR00172##

[0767] (280 mg, 0.577 mmol) was purified by SFC (column: AS (250 mm*30 mm, 5 um), gradient: 20-20% B (A=0.1% NH.sub.3/H.sub.2O, B=EtOH), flow rate: 60 mL/min) to give 3495 (20 mg, 7%) as a solid and 3496 (32 mg, 11%) as a solid.

[0768] 3495

[0769] .sup.1H NMR CDCl.sub.3 400 MHz δ 5.39-5.35 (m, 1H), 4.05-3.98 (m, 1H), 3.93-3.85 (m, 1H), 3.72-3.68 (m, 2H), 3.59-3.51 (m, 1H), 2.49 (s, 2H), 2.05-1.72 (m, 9H), 1.55-1.40 (m, 7H), 1.72-1.40 (m, 7H), 1.40-0.90 (m, 9H), 0.69 (s, 3H).

[0770] LCMS Rt=1.081 min in 2.0 min chromatography, 30-90AB_2MIN_E.M, purity 100%, MS ESI calcd. for C.sub.28H.sub.42F.sub.3O.sub.2 [M+H−H.sub.2O].sup.+ 467, found 467.

[0771] 3496

[0772] .sup.1H NMR CDCl.sub.3 400 MHz δ 5.39-5.35 (m, 1H), 4.05-3.98 (m, 1H), 3.90-3.85 (m, 1H), 3.72-3.68 (m, 1H), 3.59-3.51 (m, 1H), 2.49 (s, 2H), 2.05-1.72 (m, 10H), 1.68-1.1.60 (m, 2H), 1.52-1.25 (m, 8H), 1.25-0.92 (m, 13H), 0.69 (s, 3H).

[0773] LCMS Rt=1.095 min in 2.0 min chromatography, 30-90AB_2MIN_E.M, purity 100%, MS ESI calcd. for C.sub.28H.sub.42F.sub.3O.sub.2 [M+H−H.sub.2O].sup.+ 467, found 467.

Example 35: Synthesis of 3507

[0774] ##STR00173##

[0775] Stereochemistry assigned based on synthesis with chiral epoxide.

[0776] The experimental of intermediate ST-200-31-6 can be found in Example 33.

Synthesis of ST-200-31-5

[0777] ##STR00174##

[0778] ST-200-31-6 (210 mg, 0.43 mmol) was purified by SFC (column: AD (250 mm*30 mm, 10 um)), gradient: 20-20% B (A=0.1% NH.sub.3/H.sub.2O, B=EtOH), flow rate: 50 mL/min to give impure 3507 (100 mg, 45%) as a solid.

[0779] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.33 (m, 1H), 3.15-3.06 (m, 1H), 2.48 (s, 2H), 2.08-1.92 (m, 4H), 1.89-1.57 (m, 6H), 1.53-1.23 (m, 8H), 1.21-0.97 (m, 10H), 0.96-0.83 (m, 12H), 0.68 (s, 3H).

Synthesis of 3507

[0780] ##STR00175##

[0781] Lindlar catalyst (100 mg) was added to a solution of impure sample (100 mg, 0.21 mmol, 22,23-olefin included) in EtOAc (5 mL) under N.sub.2. The suspension was degassed under vacuum and purged with H.sub.2 for three times. Then the solution was hydrogenated under 15 psi of hydrogen at 25° C. for 4 h. The mixture was filtered through a pad of celite and washed with EtOAc (3×10 mL). The filtrate was concentrated to give a solid. .sup.1H NMR showed there was still contained 12.5% 22,23-olefin. The impure 3507 was dissolved in THF/MeOH (3/3 mL) and treated with Lindlar (100 mg) under N.sub.2. The suspension was degassed under vacuum and purged with H.sub.2 for three times. Then the solution was hydrogenated under 15 psi of hydrogen at 25° C. for 4 h. The mixture was filtered through a pad of celite and washed with THF (3×10 mL). The filtrate was concentrated and triturated from PE (5 mL) to give 3507 as a solid, which was triturated in n-hexane (5 mL) at 25° C. to give 3507 (40 mg, 40%) as a solid.

[0782] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.34 (m, 1H), 3.13-3.06 (m, 1H), 2.48 (s, 2H), 2.09-1.94 (m, 4H), 1.89-1.57 (m, 6H), 1.54-1.34 (m, 6H), 1.32-1.08 (m, 5H), 1.07-0.97 (m, 7H), 0.94 (d, J=6.4 Hz, 3H), 0.89 (s, 9H), 0.68 (s, 3H).

[0783] LCMS Rt=1.298 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.29H.sub.46F.sub.3O [M+H−H.sub.2O].sup.+ 467, found 467.

[0784] SFC_E1 Rt=3.887 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100% de.

Synthesis Confirming Stereochemistry for 3507 and 3634

[0785] ##STR00176##

[0786] To a solution of THF (0.5 mL) was added n-BuLi (0.8 mL, 2.5 M in hexane, 2 mmol), was added a solution of DD (420 mg, 0.8 mmol) in THF (2 mL) at −70° C. After stirring at −70° C. for 1 h, (R)-2-(tert-butyl)oxirane(120 mg, 1.2 mmol) in THF (0.5 mL) was added at −70° C. The mixture was stirred at −70° C. for another 1 h and warmed to 25° C. and stirred for 16 hours. The reaction mixture was quenched with sat. NH.sub.4Cl (10 mL) and extracted with EtOAc (2×5 mL). The organic layer was separated, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue (400 mg) was used directly for next step.

[0787] To a mixture of DDA (400 mg, crude) in MeOH (30 mL) was added NiCl.sub.2 (8.29 mg, 0.64 mmol) at 25° C. Then the mixture was warmed to 60° C., Mg powder (671 mg, 25.5 mmol) was added in three bathes. The reaction was quenched with HCl (1M, 10 mL), the mixture was extracted with EtOAc (2×30 mL). The combined organic layer was washed with brine (20 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash-combi (0-30% of EtOAc in PE) to give 3507 (110 mg, impure) as a solid, which was further purified by SFC ((column: AD (250 mm*30 mm, 10 um)), gradient: 30-30% B (A=0.1% NH.sub.3/H.sub.2O IPA, B=EtOH), flow rate: 50 mL/min) to give 3507 (100 mg) as a solid.

[0788] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.34 (m, 1H), 3.14-3.02 (m, 1H), 2.48 (s, 2H), 2.10-1.91 (m, 3H), 1.90-1.69 (m, 4H), 1.69-1.51 (m, 6H), 1.51-1.27 (m, 7H), 1.22-0.98 (m, 8H), 0.98-0.92 (m, 3H), 0.89 (s, 9H), 0.68 (s, 3H).

[0789] LCMS Rt=1.322 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.29H.sub.46F.sub.3O [M+H−H.sub.2O].sup.+ 467, found 467.

[0790] SFC Rt=3.804 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100% de. To a solution of 3507 (70 mg) in THF (10 mL) was added Pd(OH).sub.2/C (20%, dry, 100 mg). The mixture was stirred under H.sub.2 (50 psi) at 50° C. for 18 h. The mixture was filtered and concentrated in vacuum. The residue was purified by flash-combi (0-15% of EtOAc in PE) to give 3634 (13 mg, 19%) as a solid.

[0791] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.17-2.98 (m, 1H), 2.14-1.78 (m, 4H), 1.78-1.60 (m, 6H), 1.57-1.34 (m, 7H), 1.34-1.00 (m, 13H), 0.98 (s, 3H), 0.92 (m, 3H), 0.89 (s, 9H), 0.65 (s, 3H).

[0792] LCMS Rt=1.349 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, no MS signal.

[0793] MS ESI calcd. for C.sub.29H.sub.48F.sub.3O [M+H−H.sub.2O].sup.+ 469, found 469.

Example 36: Synthesis of 3634

[0794] ##STR00177##

[0795] The experimental procedures of intermediate 3507 can be found in Example 3.

Synthesis 3634

[0796] ##STR00178##

[0797] To a solution of 3507 (70 mg) in THF (10 mL) was added Pd(OH).sub.2/C (20%, dry, 100 mg). The mixture was stirred under H.sub.2 (50 psi) at 50° C. for 18 h. The mixture was filtered and concentrated in vacuum. The residue was purified by flash-combi (0-15% of EtOAc in PE) to give 3634 (13 mg, 19%) as a solid.

[0798] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.17-2.98 (m, 1H), 2.14-1.78 (m, 4H), 1.78-1.60 (m, 6H), 1.57-1.34 (m, 7H), 1.34-1.00 (m, 13H), 0.98 (s, 3H), 0.92 (m, 3H), 0.89 (s, 9H), 0.65 (s, 3H).

[0799] LCMS Rt=1.349 min in 2.0 min chromatography, 30-90_AB_E, purity 100%, no MS signal.

[0800] MS ESI calcd. for C.sub.29H.sub.48F.sub.3O [M+H−H.sub.2O].sup.+ 469, found 469.

Example 37: Synthesis of 3788

[0801] ##STR00179##

[0802] The experimental of intermediate ST-200-31-4 can be found in Example 33.

Synthesis of 3788

[0803] ##STR00180##

[0804] Pd(OH).sub.2/C (100 mg) was added to a solution of ST-200-31-4 (60 mg, 0.12 mmol) in THF/MeOH (5 mL/5 mL) and the mixture was degassed and back-filled with H.sub.2 three times. Next, the reaction was stirred at 50° C. under 50 psi of H.sub.2 for 16 h. The reaction mixture was filtered through a pad of celite washed with EtOAc (100 mL). The filtrate was concentrated to give impure ST-200-31-3B as a solid. To a solution of the impure ST-200-31-4 in THF/MeOH (3 mL/3 mL) was added Pd(OH).sub.2/C (50 mg) and the mixture was degassed and back-filled with H.sub.2 for 3 times. After that, the reaction was stirred at 50° C. under 50 psi of H.sub.2 for 72 h. The reaction mixture was filtered through a pad of celite washed with EtOAc (100 mL). The filtrate was concentrated to give 40 mg of crude product, which was triturated in n-hexane (2×3 mL) to give 3788 (7 mg, 17%) as a solid.

[0805] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.19-3.08 (m, 1H), 2.13-1.81 (m, 4H), 1.77-1.58 (m, 4H), 1.54-1.35 (m, 9H), 1.34-1.01 (m, 13H), 1.01-0.96 (m, 3H), 0.94-0.86 (m, 12H), 0.66 (s, 3H).

[0806] LCMS Rt=1.313 min in 2.0 min chromatography, 30-90AB_2MIN_E, purity 98%, MS ESI calcd. for C.sub.29H.sub.48F.sub.3O [M+H−H.sub.2O].sup.+ 469, found 469.

Example 38: Synthesis of 3877 and 3886

[0807] ##STR00181##

[0808] Stereochemistry for 3877 is shown below; assigned by NMR.

Synthesis of ST-200-74-5_1

[0809] ##STR00182##

[0810] Me.sub.3SI (4.71 g, 23.1 mmol) was added to a suspension of t-BuOK (3.98 g, 35.6 mmol) in THF (40 mL) under N.sub.2 at 35° C. After stirring at 35° C. for 30 mins, a solution of ST-200-74-5_1 (2 g, 17.8 mmol) was added dropwise at 35° C. The mixture was stirred at 35° C. for 16 hrs, quenched with sat. NH.sub.4Cl (50 mL) and extracted with EtOAc (3×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give ST-200-74-5_2 (1.8 g, crude) as liquid which was used directly for next step.

Synthesis of ST-200-74-5_3

[0811] ##STR00183##

[0812] n-BuLi (0.948 mL, 2.5 M in hexane, 2.37 mmol) was added to THF (5 mL). A solution of ST-200-CF3_6C (500 mg, 0.949 mmol) in THF (15 mL) was added at −70° C. After stirring at −70° C. for 1 h, 6-methyl-1-oxaspiro[2.5]octane(358 mg, 2.84 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 hour, then warmed to 15° C. and stirred for 16 hrs. After quenching with NH.sub.4Cl (50 mL), the mixture was extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, concentrated, and purified by combi-flash (0-20% of EtOAc in PE) to give ST-200-74-5_3 (350 mg, crude) as a solid, which was used directly for the next step.

Synthesis of 3877

[0813] ##STR00184##

[0814] A solution of ST-200-74-5_3 (350 mg, 0.536 mmol) in MeOH (30 mL) was heated at 65° C. Mg powder (513 mg, 21.4 mmol) was added in one portion at 65° C. The mixture was refluxed at 65° C. for 1 h. The mixture was quenched with HCl (40 mL, 2N) until the reaction became clear and extracted with DCM (2×30 mL). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by silica gel chromatography (0-12% of EtOAc in PE) to give 3877 (12 mg, 4%) as a solid.

[0815] 3877:

[0816] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.11-1.90 (m, 3H), 1.89-1.74 (m, 2H), 1.73-1.58 (m, 5H), 1.53-1.43 (m, 6H), 1.42-1.19 (m, 14H), 1.18-0.96 (m, 7H), 0.96-0.80 (m, 10H), 0.74-0.60 (m, 4H).

[0817] LCMS Rt=1.728 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%.

[0818] MS ESI Scan (2.939-3.092 min, 10 scans) Frag=50.0 V, 80-100_1_4min.m, MS ESI calcd. For C.sub.31H.sub.51F3Q.sub.2Na [M+Na].sup.+ 535, found 535.

##STR00185##

Synthesis of ST-200-096-011A/B

[0819] ##STR00186##

[0820] To a solution of ST-200-74-5_3 (700 mg, 1.07 mmol) in MeOH (40 mL) was added NiCl.sub.2 (27.6 mg, 0.214 mmol) and Mg powder (1.02 g, 41.8 mmol) at 65° C. in one portion. The mixture was stirred at 65° C. for 10 minutes. Another Mg powder (513 mg, 22.3 mmol) was added in one portion. After stirring at 65° C. for 10 minutes, the mixture was quenched with HCl (200 mL, 1N) and extracted with EtOAc (3×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-15% of EtOAc in PE) to give ST-200-096-011A (63 mg, 11%, Peak 1) and ST-200-096-011B (114 mg, 20%, Peak 2) as a solid. 3877

[0821] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.09-1.93 (m, 3H), 1.90-1.76 (m, 2H), 1.73-1.57 (m, 8H), 1.51-1.34 (m, 8H), 1.33-1.18 (m, 6H), 1.17-0.98 (m, 8H), 0.97-0.87 (m, 7H), 0.84 (s, 3H), 0.73-0.63 (m, 4H).

[0822] LCMS Rt=1.391 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%.

[0823] MS ESI Scan (1.955-2.16 min, 8 scans) Frag=50.0 V, 80-100_1_4min.m, MS ESI calcd. For C.sub.31H.sub.51F.sub.3O.sub.2Na [M+Na].sup.+ 535, found 535.

[0824] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.09-2.00 (m, 2H), 1.99-1.89 (m, 1H), 1.87-1.76 (m, 2H), 1.71-1.61 (m, 3H), 1.55-0.42 (m, 10H), 1.41-1.19 (m, 13H), 1.14-0.96 (m, 6H), 0.95-0.86 (m, 7H), 0.84 (s, 3H), 0.72-0.62 (m, 4H).

[0825] LCMS Rt=1.450 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%.

[0826] MS ESI Scan (1.938-2.617 min, 9 scans) Frag=50.0 V, 80-100_1_4min.m, MS ESI calcd. For C.sub.31H.sub.51F.sub.3O.sub.2Na [M+Na].sup.+ 535, found 535.

Example 39: Synthesis of 3983

[0827] ##STR00187##

[0828] See Example 5 for synthesis of ST-200-CF3_6C.

Synthesis of ST-310-15-2_2

[0829] ##STR00188##

[0830] A solution of Me.sub.3SI (13.6 g, 66.7 mmol) and t-BuOK (17.8 mL, 5M in THF, 89.0 mmol) in DMSO (100 mL) was stirred and heated at 25° C. for 30 min under N.sub.2. Cycloheptanone (5 g, 44.5 mmol) was added to the reaction mixture and stirred at 25° C. for 3 hrs. The reaction was treated with water (300 mL), extracted with EtOAc (2×100 mL). The combined organic phase was washed with water (2×300 mL), brine (2×300 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuum to afford ST-200-74-5_2 (4 g, 71%) as a liquid.

[0831] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.59 (s, 2H), 1.72-1.50 (m, 12H).

Synthesis of ST-310-15-2_3

[0832] ##STR00189##

[0833] Added n-BuLi (0.568 mL, 1.42 mmol, 2.5 M in hexane) was added to a solution of ST-200-CF3_6C (300 mg, 0.569 mmol) in THF (3 mL) at −70° C. under N.sub.2. After cooling to −70° C., 1-oxaspiro [2.6] nonane (107 mg, 0.853 mmol) was added. The reaction was allowed to warm to 25° C. and was stirred for 12 hours at 25° C. The reaction was quenched with NH.sub.4Cl (10 mL, sat. aq.), water (50 mL) and extracted with EtOAc (3×10 mL). The combined organic phase was concentrated to give a residue, which was purified by silica gel chromatography (PE/EtOAc=10/1-5/1) to give compound ST-200-74-6_3 (200 mg, impure) as an oil. The crude mixture was used directly for the next step.

Synthesis of 3983

[0834] ##STR00190##

[0835] A solution of ST-200-74-6_3 (200 mg, 306 umol) in MeOH (50 mL) was heated to 60° C. Mg powder (371 mg, 15.3 mmol) was added in four portions at 60° C. After stirring at 60° C. for 1 h, the mixture was quenched with HCl (50 mL, 2 M) until the reaction became clear and extracted with EtOAc (2×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated and purified by flash column (0-40% of EtOAc in PE) to give 3983 (15 mg, 12%) as a solid.

[0836] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.12-2.00 (s, 1H), 1.99-1.92 (m, 2H), 1.89-1.77 (m, 2H), 1.74-1.57 (m, 10H), 1.57-1.52 (m, 6H), 1.41-1.17 (m, 13H), 1.16-0.95 (m, 7H), 0.94-0.82 (m, 6H), 0.72-0.63 (m, 4H).

[0837] LCMS Rt=0.690 min in 2 min chromatography, 30-90 AB, purity 100%.

[0838] HRMS ESI calcd. for C.sub.31H.sub.50F.sub.3O [M+H−H.sub.2O].sup.+ 495, found 495.

Example 40: Synthesis of 4023

[0839] ##STR00191##

[0840] The synthesis of ST-200-CF3_6C can be found in Example 5.

Synthesis of M-1-19_3

[0841] ##STR00192##

[0842] n-BuLi (2.5 M, 1.42 mmol, 0.568 mL) was added to THF (2 mL) under N.sub.2 at −70° C. Next, a suspension of ST-200-CF3_6C (300 mg, 0.569 mmol) in THF (2 mL) was added drop-wise to give a suspension. After stirring at −70° C. for 30 min, a solution of 1-oxaspiro [2.5] octane (126 mg, 1.13 mmol) was added. The reaction was stirred at stirred at 25° C. for 16 hours. The mixture was poured into ice-water (20 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to afford M-1-19_3 (280 mg, crude) as a solid, which was used directly for the next step.

Synthesis of 4023

[0843] ##STR00193##

[0844] Mg (212 mg, 8.75 mmol) and NiCl.sub.2 (11.3 mg, 0.088 mmol) were added to a solution of M-1-19_3 (280 mg, 0.438 mmol) in 20 mL of dry methanol at 25° C. The mixture was stirred at 50° C. for 1 h. The mixture was quenched by 2M HCl (50 mL) at 10° C. until solid was dissolved. The mixture was extracted with EtOAc (50 mL). The organic layers were washed with sat. NaHCO.sub.3 (50 mL), brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel column eluted with PE/EtOAc=10/1 to afford 4023 (38 mg, 14%) as a solid.

[0845] .sup.1H NMR (400 MHz, CDCl3) δ 2.08-2.02 (m, 1H), 2.01-1.94 (m, 2H), 1.89-1.78 (m, 2H), 1.71-1.59 (m, 5H), 1.53-1.32 (m, 13H), 1.30-1.05 (m, 13H), 1.03-0.83 (m, 9H), 0.72-0.85 (m, 4H).

[0846] MS ESI calcd. for C.sub.30H.sub.48F.sub.3O [M+H−H.sub.2O].sup.+ 481, found 481.

Example 41: Synthesis of 4155 and 4156

[0847] ##STR00194##

[0848] See Example 11 for synthesis of ST-200-74-1_5.

Synthesis of ST-200-74-1_6

[0849] ##STR00195##

[0850] cyclohexylmagnesium chloride (2.55 mL, 5.1 mmol, 2M in THF) was added dropwise to a solution of ST-200-74-1_5 (440 mg, 1.02 mmol) in THF (10 mL) at 0° C. The mixture was stirred at 25° C. for 1 h. The reaction mixture was quenched with water (20 mL) and extracted with EtOAc (2×20 mL). The combined organic phase was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum. The residue was purified by silica gel column eluted with (PE/EtOAc=5/1) to afford ST-200-74-1_6 (400 mg, 77%) as a solid.

[0851] .sup.1H NMR (400 MHz, CDCl3) δ 3.32-3.28 (m, 1H), 2.28-2.23 (m, 1H), 2.08-2.02 (m, 1H), 1.98-1.79 (m, 6H), 1.58-1.34 (m, 15H), 1.30-1.00 (m, 15H), 0.95-0.83 (m, 8H), 0.72-0.65 (m, 4H).

Synthesis of ST-200-74-1_7

[0852] ##STR00196##

[0853] Benzoyl chloride (164 mg, 1.17 mmol) was added to a solution of ST-200-74-1_6 (400 mg, 0.78 mmol) in Pyridine (4 mL) at 25° C. The mixture was stirred at 25° C. for 12 hrs. The mixture was poured into water (50 mL) and extracted with ethyl acetate (2×50 mL). The combined organic layers was washed with brine (20 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue was purified by silica gel column eluted with (PE/EtOAc=10/1) to afford ST-200-74-1_7 (315 mg, 65%) as an oil.

[0854] .sup.1H NMR (400 MHz, CDCl3) δ 8.06-8.04 (m, 2H), 7.62-7.50 (m, 1H), 7.46-7.43 (m, 2H), 4.98-4.90 (m, 1H), 2.07-2.04 (m, 1H), 1.95-1.92 (m, 1H), 1.82-1.55 (m, 10H), 1.54-1.30 (m, 10H), 1.28-1.05 (m, 13H), 0.99-0.93 (m, 10H), 0.67-0.61 (m, 4H).

Synthesis of ST-200-74-1_8A, 8B

[0855] ##STR00197##

[0856] ST-200-74-1_7 (315 mg) was purified by SFC (Column: AD (250 mm*30 mm, 5 um), Condition: 0.1% NH.sub.3.H.sub.2O, IPA, Gradient: from 40% to 40%, FlowRate (ml/min): 60 mL/min, 25° C.) to afford ST-200-74-1_8A (115 mg, 37%) and ST-200-74-1_8B (108 mg, 35%) as a solid.

[0857] ST-200-74-1_8A

[0858] .sup.1H NMR (400 MHz, CDCl3) δ 8.06-8.03 (m, 2H), 7.58-7.53 (m, 1H), 7.46-7.43 (m, 2H), 4.98-4.90 (m, 1H), 2.07-2.02 (m, 1H), 1.96-1.91 (m, 2H), 1.84-1.62 (m, 12H), 1.53-1.24 (m, 11H), 1.22-0.96 (m, 12H), 0.94-0.83 (m, 7H), 0.70-0.64 (m, 1H), 0.61 (s, 3H).

[0859] ST-200-74-1_8B

[0860] .sup.1H NMR (400 MHz, CDCl3) δ 8.06-8.03 (m, 2H), 7.58-7.53 (m, 1H), 7.46-7.43 (m, 2H), 4.98-4.90 (m, 1H), 2.07-1.91 (m, 3H), 1.84-1.69 (m, 7H), 1.67-1.48 (m, 9H), 1.43-1.32 (m, 4H), 1.30-1.02 (m, 13H), 1.01-0.84 (m, 9H), 0.7-0.62 (m, 4H).

Synthesis of 4156

[0861] ##STR00198##

[0862] KOH (52.1 mg, 0.93 mmol) was added to a solution of ST-200-74-1_8A (115 mg, 0.186 mmol) in THF (2 mL), MeOH (1 mL) and water (1 mL). The mixture was stirred at 60° C. for 16 hrs. The mixture was poured into water (20 mL) and extracted with EtOAc (2×40 mL). The combined organic layer was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (PE/EtOAc=5/l to 3/1) to give 4156 (56 mg, 59%) as a solid.

[0863] .sup.1H NMR (400 MHz, CDCl3) δ 3.31-3.27 (m, 1H), 2.08-2.02 (m, 1H), 1.98-1.93 (m, 2H), 1.84-1.72 (m, 5H), 1.70-1.60 (m, 7H), 1.51-1.46 (m, 2H), 1.42-1.36 (m, 3H), 1.34-1.11 (m, 13H), 1.06-0.85 (m, 13H), 0.72-0.65 (m, 4H).

[0864] MS ESI calcd. for C.sub.31H.sub.50F.sub.3O [M+H−H.sub.2O].sup.+ 495, found 495.

Synthesis of 4155

[0865] ##STR00199##

[0866] KOH (49 mg, 0.875 mmol) was added to a solution of ST-200-74-1_8B (108 mg, 0.175 mmol) in THF (2 mL), MeOH (1 mL) and water (1 mL). The mixture was stirred at 60° C. for 16 hrs. The mixture was poured into water (20 mL) and extracted with EtOAc (2×40 mL). The combined organic layer was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (PE/EtOAc=5/l to 3/1) to give 4155 (56 mg, 62%) as a solid.

[0867] .sup.1H NMR (400 MHz, CDCl3) δ 3.31-3.27 (m, 1H), 2.08-2.02 (m, 1H), 1.98-1.93 (m, 2H), 1.84-1.72 (m, 5H), 1.70-1.60 (m, 6H), 1.51-1.34 (m, 9H), 1.31-0.97 (m, 17H), 0.95-0.85 (m, 6H), 0.72-0.65 (m, 4H).

[0868] MS ESI calcd. for C.sub.31H.sub.51F.sub.3O.sub.2Na [M+Na].sup.+ 535, found 535.

Synthesis Confirming Stereochemistry of 4155

[0869] ##STR00200## ##STR00201##

[0870] To a suspension of C.sub.3H.sub.9IS (117 g, 578 mmol) in THF (300 mL) was added a solution of t-BuOK (99.6 g, 890 mmol) in THF (400 mL) slowly under N.sub.2 at 30° C. The suspension was stirred at 30° C. for 30 min. Then ST-200-096-008_1 (50 g, 445 mmol) in 100 ml of THF was added dropwise to the mixture at 0° C. After stirring at 30° C. for 16 hrs, the mixture was poured into sat. NH.sub.4Cl (600 mL) and extracted with EtOAc (2×200 mL). The combined organic phase was washed with brine (400 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated at 40° C. under reduced pressure to give ST-200-096-008_2 (55 g, crude) as a liquid.

[0871] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.75-2.65 (m, 2H), 2.55-2.50 (m, 1H), 1.90-1.80 (m, 1H), 1.78-1.58 (m, 4H), 1.30-1.00 (m, 6H)

[0872] To a suspension of C.sub.3H.sub.9IS (117 g, 578 mmol) in THF (300 mL) was added a solution of t-BuOK (99.6 g, 890 mmol) in THF (400 mL) slowly under N.sub.2 at 30° C. The suspension was stirred at 30° C. for 30 min. Then ST-200-096-008_1 (50 g, 445 mmol) in 100 ml of THF was added dropwise to the mixture at 0° C. After stirring at 30° C. for 16 hrs, the mixture was poured into sat. NH.sub.4Cl (600 mL) and extracted with EtOAc (2×200 mL). The combined organic phase was washed with brine (400 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated at 40° C. under reduced pressure to give ST-200-096-008_2 (55 g, crude) as a liquid.

[0873] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.75-2.65 (m, 2H), 2.55-2.50 (m, 1H), 1.90-1.80 (m, 1H), 1.78-1.58 (m, 4H), 1.30-1.00 (m, 6H)

[0874] To a solution of R,R-cat (190 mg, 0.316 mmol) in toluene (3 mL) was added AcOH (189 mg, 3.16 mmol). The mixture was stirred at 25° C. under air for 30 min and concentrated in vacuum to leave a crude brown solid. The resulting catalyst residue was dissolved in 2-cyclohexyloxirane (10 g, 79.2 mmol) at 25° C. The reaction flask was cooled to 0° C., and H.sub.2O (783 g, 43.5 mmol) was added dropwise over 5 min. After stirring at 25° C. for 24 hrs, ((2R)-2-cyclohexyloxirane (2 g, 15.8 mmol, 20.0%) was isolated by distillation from the reaction mixture. To a solution of ST-200-096-008_3 (50 mg, 0.396 mmol) and TEA (39.9 mg, 0.396 mmol) in MeOH (3 mL) was added naphthalene-2-thiol (63.4 mg, 0.396 mmol) at 25° C. After stirring at 25° C. for 2 hrs, the ee % of (2R)-2-cyclohexyloxirane was determined to be 82.7% by chiral HPLC.

[0875] SFC Peak 1: Rt=2.033 min in 10 min chromatography, Chiralpak AD-3 100×4.6 mm I.D., 3 μm, 82.7% ee.

[0876] To THF (1 mL) was added BuLi (1.12 mL, 2.5 M in hexane, 2.82 mmol). A solution of ST-200-CF3_6C (600 mg, 1.13 mmol) in THF (6 mL) was added at −70° C. The mixture was stirred at −70° C. for 1 h. (2R)-2-cyclohexyloxirane (213 mg, 1.69 mmol) was added at −70° C. After stirring at 30° C. and stirred for 16 hrs, the reaction mixture was quenched with sat. NH.sub.4Cl (50 mL) and extracted with EtOAc (2×30 mL). The combined organic layer were washed with NH.sub.4Cl (50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated to give crude product (700 mg) as a foam solid, which was used for next step directly.

[0877] To a solution of ST-200-096-008_4 (700 mg, 1.07 mmol) in MeOH (60 mL) was added NiCl.sub.2 (27.6 mg, 0.214 mmol) and Mg powder (1.02 g, 42.8 mmol) at 65° C. in one portion. The mixture was stirred at 65° C. for 10 minutes. Another Mg powder (513 g, 21.4 mmol) was added in one portion. After stirring at 65° C. for 10 minutes, the mixture was quenched with HCl (120 mL, 1N) and extracted with EtOAc (3×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-15% of EtOAc in PE) to give ST-200-096-008 (300 mg, 55%) as a solid.

[0878] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.35-3.25 (m, 1H), 2.10-2.01 (m, 1H), 2.00-1.91 (m, 1H), 1.78-1.72 (m, 5H), 1.71-1.58 (m, 9H), 1.56-1.10 (m, 19H), 1.09-0.98 (m, 4H), 0.97-0.86 (m, 4H), 0.84 (s, 3H), 0.72-0.60 (m, 4H)

[0879] To a solution of ST-200-096-008 (300 mg, 0.585 mmol) in pyridine (5 mL) was added benzoyl chloride (164 mg, 1.17 mmol) at 25° C. After stirring at 25° C. for 12 hours, the mixture was poured into water (50 mL) and extracted with ethyl acetate (2×30 mL). The combined organic layers was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue was purified by silica gel column eluted with (PE/EtOAc=10/1) to afford ST-200-096-008_5 (300 mg) as a solid, which was separated by SFC (column: AD (250 mm*30 mm, 5 um), gradient: 35-35% B (A=0.05% NH.sub.3/H.sub.2O, B=MeOH), flow rate: n/a mL/min) to give 100% de product (190 mg, 52% yield for 2 steps) as a solid.

[0880] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.08-8.01 (m, 2H), 7.58-7.52 (m, 1H), 7.58-7.40 (m, 2H), 4.98-4.90 (m, 1H), 2.19-2.10 (m, 2H), 1.97-1.89 (m, 1H), 1.83-1.58 (m, 12H), 1.56-1.35 (m, 8H), 1.34-0.95 (m, 15H), 0.94-0.89 (m, 3H), 0.88-0.79 (m, 4H), 0.70-0.59 (m, 4H).

[0881] SFC Peak 1: Rt=5.105 min and Peak 2 Rt=5.644 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML (“Column: Chiralpak AD-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO2 B: iso-propanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.”).

[0882] SFC Peak 1: Rt=5.313 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100.0% de.

[0883] To a solution of ST-200-096-008_5 (190 mg, 0.308 mmol) in THF (2 mL) and MeOH (4 mL) and water (1 mL) was added NaOH (246 mg, 6.16 mmol). After stirring at 50° C. for 16 hrs, the mixture was poured into water (20 mL) and extracted with EtOAc (2×20 mL). The combined organic layer was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (PE/EtOAc=5/l to 3/1) to give ST-200-096-008 (126 mg, 80%) as a solid.

[0884] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.35-3.25 (m, 1H), 2.10-2.01 (m, 2H), 2.00-1.91 (m, 1H), 1.78-1.72 (m, 5H), 1.71-1.58 (m, 5H), 1.56-1.50 (m, 5H), 1.49-1.18 (m, 13H), 1.17-0.95 (m, 8H), 0.94-0.86 (m, 4H), 0.84 (s, 3H), 0.70-0.61 (m, 4H).

[0885] LCMS Rt=1.389 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%.

[0886] MS ESI Scan (1.981-2.144 min, 11 scans) Frag=50.0 V, 80-100_1_4min.m, MS ESI calcd. For C.sub.31H.sub.51F.sub.3O.sub.2Na [M+Na].sup.+ 535, found 535.

Example 42: Synthesis of 4258 and 4259

[0887] ##STR00202##

[0888] The stereochemistry for 4259 was confirmed by Xray data. The synthesis of ST-200-CF3_6C can be found in Example 5.

Synthesis of M-1-14_2

[0889] ##STR00203##

[0890] To a suspension of Me3SI (1.88 g, 9.26 mmol) in THF (10 mL) was slowly added a solution of t-BuOK (1.59 g, 14.2 mmol) in THF (5 mL) under N.sub.2 at 15° C. The suspension was stirred at 15° C. for 30 min. Then a solution of M-1-14_1 (1 g, 7.13 mmol) in 5 ml of THF was added dropwise to the mixture at 0° C. After addition, the mixture was stirred at 15° C. for 16 hrs. The reaction was quenched with sat. NH.sub.4Cl (60 mL) and extracted with MTBE (3×30 mL). The combined organic phase was washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated at 40° C. under reduced pressure to give M-1-14_2 (1 g, crude) as a liquid.

[0891] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.65-2.55 (m, 2H), 1.90-1.80 (m, 3H), 1.74-1.66 (m, 1H), 1.60-1.46 (m, 1H), 1.42-1.22 (m, 2H), 1.21-1.10 (m, 3H), 0.92-0.80 (m, 6H).

Synthesis of M-1-14_3

[0892] ##STR00204##

[0893] To THF (0.5 mL) was added n-BuLi (0.568 mL, 2.5 M in hexane, 1.42 mmol) at −70° C. A solution of ST-200-CF3_6C (300 mg, 0.569 mmol) in THF (2.5 mL) was added dropwise at −70° C. After stirring at −70° C. for 1 h, 6-isopropyl-1-oxaspiro[2.5]octane (131 mg, 0.853 mmol) was added. The mixture was stirred at −70° C. for another 1 h. Then the reaction mixture was warmed to 15° C. and stirred for 16 hrs. The mixture was quenched with NH.sub.4Cl (50 mL, sat. aq.) and extracted with EtOAc (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, and concentrated to give M-1-14_3 (350 mg, crude) as a solid, which was used for next step directly.

Synthesis of 4259 and 4258

[0894] ##STR00205##

[0895] To a solution of M-1-14_3 (350 mg, 0.513 mmol) in MeOH (30 mL) was added NiCl.sub.2 (13.2 mg, 0.102 mmol) and Mg powder (492 mg, 20.5 mmol) at 65° C. in one portion. After stirring at 65° C. for 10 minutes, another Mg powder (244 mg, 10.2 mmol) was added in one portion. The mixture was stirred at 65° C. for another 10 minutes. The mixture was quenched with HCl (50 mL, 2N) until the reaction became clear and extracted with EtOAc (3×20 mL). The combined organic layer was washed with sat. NH.sub.4Cl (50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by silica gel chromatography (0-15% of EtOAc in PE) to give 4258 (24 mg, 8.6%, 4258) and 4259 (50 mg, 18%, 4259) as a solid.

[0896] 4258

[0897] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.10-2.01 (m, 1H), 2.00-1.92 (m, 2H), 1.89-1.56 (m, 12H), 1.51-1.33 (m, 8H), 1.32-1.18 (m, 6H), 1.17-0.98 (m, 9H), 0.98-0.89 (m, 4H), 0.88-0.83 (m, 9H), 0.74-0.63 (m, 4H).

[0898] HPLC Rt=7.214 min in 10.0 min chromatography, 50-100AB_E, purity 98.8%.

[0899] MS 80-100_1_4min.m, MS ESI calcd. for C.sub.33H.sub.54F.sub.3O [M+H−H.sub.2O].sup.+ 523, found 523.

[0900] 4259

[0901] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.10-2.01 (m, 1H), 1.98-1.91 (m, 2H), 1.89-1.75 (m, 2H), 1.73-1.569 (m, 6H), 1.55-1.33 (m, 11H), 1.32-1.14 (m, 10H), 1.13-0.92 (m, 7H), 0.91-0.83 (m, 12H), 0.73-0.62 (m, 4H).

[0902] LCMS Rt=1.816 min in 2.0 min chromatography, 30-90AB_E, purity 100%.

[0903] MS 80-100DB_1_4min.m, MS ESI calcd. for C33H54F3O [M+H−H2O].sup.+ 523, found 523.

Example 43: Synthesis of 4360

[0904] ##STR00206##

[0905] To a suspension of C.sub.3H.sub.9IS (17.1 g, 84.2 mmol) in THF (100 mL) was added a solution of t-BuOK (14.4 g, 129 mmol) in THF (50 mL) slowly under N.sub.2 at 15° C. The suspension was stirred at 15° C. for 30 min. Then M-1-13_1 (10 g, 64.8 mmol) in 50 ml of THF was added dropwise to the mixture at 0° C. After addition, the mixture was stirred at 15° C. for 16 hrs. The mixture was poured into sat. NH.sub.4Cl (300 mL) and extracted with EtOAc (3×100 mL). The combined organic phase was washed with brine (300 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated at 40° C. under reduced pressure to give M-1-13_2 (9 g, crude) as a liquid.

[0906] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.65-2.55 (m, 2H), 1.92-1.72 (m, 4H), 1.42-1.22 (m, 3H), 1.21-1.01 (m, 2H), 0.88 (s, 9H).

Synthesis of M-1-14_3

[0907] ##STR00207##

[0908] To THF (0.5 mL) was added n-BuLi (0.568 mL, 2.5 M in hexane, 1.42 mmol). A solution of ST-200-CF3_6C (300 mg, 0.569 mmol) in THF (2.5 mL) was added at −70° C. After stirring at −70° C. for 1 h, 6-(tert-butyl)-1-oxaspiro[2.5]octane (143 mg, 0.853 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 h, then warmed to 15° C. and stirred for 16 hrs. The reaction was quenched with sat. NH.sub.4Cl (50 mL) and extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give M-1-13_3 (350 mg, crude) as a solid, which was used for next step directly.

Synthesis of 4360

[0909] ##STR00208##

[0910] To a solution of M-1-13_3 (350 mg, 0.503 mmol) in MeOH (30 mL) was added NiCl.sub.2 (12.8 mg, 0.100 mmol) and Mg powder (482 mg, 20.1 mmol) at 65° C. in one portion. The mixture was stirred at 65° C. for 10 minutes. Then another batch of Mg powder (240 mg, 10.0 mmol) was added at 65° C. in one portion. The mixture was stirred at 65° C. for another 10 minutes. The reaction was quenched with HCl (50 mL, 2N) until the reaction became clear and extracted with EtOAc (3×20 mL). The combined organic layer was washed with sat. NH.sub.4Cl (50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by silica gel chromatography (0-15% of EtOAc in PE) to give 4360 (30 mg, 11%) as a solid.

[0911] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.10-2.02 (m, 1H), 1.99-1.91 (m, 2H), 1.87-1.77 (m, 2H), 1.72-1.56 (m, 7H), 1.53-1.43 (m, 4H), 1.42-1.19 (m, 13H), 1.19-0.97 (m, 7H), 0.96-0.88 (m, 5H), 0.88-0.82 (m, 12H), 0.72-0.64 (m, 4H).

[0912] HPLC Rt=7.685 min in 10.0 min chromatography, 50-100AB_E, purity 98.3%.

[0913] MS 80-100_1_4min.m, MS ESI calcd. for C.sub.34H.sub.56F.sub.3O [M+H−H.sub.2O].sup.+ 537, found 537.

Example 44: Synthesis of 4475 and 4476

[0914] ##STR00209##

[0915] The stereochemistry for 4476 was confirmed by X-ray data.

[0916] The synthesis of ST-200-CF3_6C can be found in Example 5.

Synthesis of M-1-16_2

[0917] ##STR00210##

[0918] To a suspension of Me.sub.3SI (2.08 g, 10.2 mmol) in THF (10 mL) was added a solution of t-BuOK (1.76 g, 15.8 mmol) in THF (5 mL) slowly under N.sub.2 at 15° C. The suspension was stirred at 15° C. for 30 mins. Then M-1-16_1 (1 g, 7.13 mmol) in THF (5 ml) was added dropwise to the mixture at 0° C. After the addition, the mixture was stirred at 15° C. for 16 hrs. The mixture was poured into sat. NH.sub.4Cl (60 mL), extracted with EtOAc (3×30 mL). The combined organic phase was washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated at 40° C. under reduced pressure to give M-1-16_2 (800 mg, crude) as a liquid.

[0919] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.67-2.53 (m, 2H), 1.94-1.70 (m, 4H), 1.64-1.00 (m, 8H), 0.97-0.83 (m, 3H).

Synthesis of M-1-16_3

[0920] ##STR00211##

[0921] n-BuLi (0.756 mL, 2.5 M in hexane, 1.89 mmol) was diluted with THF (0.5 mL). A solution of ST-200-CF3_6C (400 mg, 0.759 mmol) in THF (2.5 mL) was added dropwise at −70° C. The mixture was stirred at −70° C. for 1 h. 6-methoxy-1-oxaspiro[2.5]octane (158 mg, 1.13 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 h. The mixture was warmed to 15° C. and stirred for 16 hrs. To the mixture was added NH.sub.4Cl (15 mL.). The mixture was extracted with EtOAc (3×15 mL). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated to give M-1-12_3 (350 mg, crude) as an oil, which was used for the next step directly.

Synthesis of 4476 and 4475

[0922] ##STR00212##

[0923] To a solution of M-1-16_3 (350 mg, 0.524 mmol) in MeOH (30 mL) was added NiCl.sub.2 (20 mg, 0.157 mmol) and Mg powder (626 mg, 26.1 mmol) at 65° C. in one portion. The mixture was stirred at 65° C. for 1 hr. The mixture was quenched with HCl (30 mL, 2N) until the reaction became clear and extracted with EtOAc (3×20 mL). The combined organic layer was washed with sat. NH.sub.4Cl (50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by silica gel chromatography (0-15% of EtOAc in PE) to give 34 mg, 12.3%, 4476 as a solid and 57 mg, 21%, 4475 as a solid.

[0924] 4476

[0925] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.11-2.02 (m, 1H), 2.01-1.92 (m, 2H), 1.90-1.76 (m, 3H), 1.75-1.60 (m, 7H), 1.59-1.55 (m, 2H), 1.52-1.43 (m, 3H), 1.42-1.19 (m, 13H), 1.17-0.96 (m, 9H), 0.94-0.90 (m, 3H), 0.89-0.83 (m, 6H), 0.74-0.62 (m, 4H).

[0926] LCMS Rt=1.591 min in 2 min chromatography, 30-90 AB, purity 100%, no MS signal.

[0927] MS: MS ESI calcd. For C.sub.32H.sub.52F.sub.3O [M+H−H.sub.2O].sup.+ 509, found 509.

[0928] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.11-2.02 (m, 1H), 2.00-1.91 (m, 2H), 1.89-1.75 (m, 2H), 1.73-1.52 (m, 10H), 1.51-1.33 (m, 7H), 1.32-1.18 (m, 10H), 1.17-0.96 (m, 7H), 0.95-0.81 (m, 10H), 0.73-0.62 (m, 4H).

[0929] LCMS Rt=1.679 min in 2 min chromatography, 30-90 AB, purity 100%, no MS signal.

[0930] MS: MS ESI calcd. For C.sub.32H.sub.52F.sub.3O [M+H−H.sub.2O].sup.+ 509, found 509.

Example 45: Synthesis of 4555 and 4585

[0931] ##STR00213##

[0932] The synthesis of ST-200-CF3_6C can be found in Example 5.

Synthesis of M-1-12_2

[0933] ##STR00214##

[0934] A solution of t-BuOK (1.74 g, 15.6 mmol) in THF (5 mL) was slowly added to a suspension of C.sub.3H.sub.9IS (2.06 g, 10.1 mmol) in THF (10 mL) under N.sub.2 at 15° C. The suspension was stirred at 15° C. for 30 min. Then M-1-12_1 (1 g, 7.80 mmol) in 5 ml of THF was added dropwise to the mixture at 0° C. After stirring at 15° C. for 16 hours, the mixture was poured into sat. NH.sub.4Cl (60 mL) and extracted with EtOAc (3×30 mL). The combined organic phase was washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated at 40° C. under reduced pressure to give M-1-12_2 (1 g, crude) as liquid.

[0935] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.48-3.25 (m, 4H), 2.68-2.26 (m, 2H), 1.98-1.85 (m, 2H), 1.84-1.66 (m, 3H), 1.65-1.55 (m, 2H), 1.48-1.41 (m, 1H).

Synthesis of A-1-12_3

[0936] ##STR00215##

[0937] n-BuLi (0.568 mL, 2.5 M in hexane, 1.42 mmol) was added to THF (0.5 mL). A solution of ST-200-CF3_6C (300 mg, 0.569 mmol) in THF (2.5 mL) was added at −70° C. After stirring at −70° C. for 1 h, 6-methoxy-1-oxaspiro[2.5]octane (121 mg, 0.853 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 h. The mixture was warmed to 15° C. and stirred for 16 hrs. The reaction mixture was quenched with NH.sub.4Cl (50 mL, sat. aq.) and extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give M-1-12_3 (350 mg, crude) as a solid, which was used for next step directly.

Synthesis of 4555

[0938] ##STR00216##

[0939] NiCl.sub.2 (13.4 mg, 0.104 mmol) and Mg powder (501 mg, 20.9 mmol) were added in one portion to a solution of M-1-12_3 (350 mg, 0.523 mmol) in MeOH (40 mL) at 65° C. The mixture was stirred at 65° C. for 10 minutes. Another portion of Mg powder (250 mg, 10.4 mmol) was added. After stirring at 65° C. for 10 minutes, the mixture was quenched with HCl (60 mL, 1N) and extracted with EtOAc (3×20 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-15% of EtOAc in impure M-12(100 mg) as a solid, which was hydrogenated (dry Pd(OH).sub.2 (40 mg), MeOH (10 mL), 50° C., 50si for 48 hrs). The suspension was filtered and the filtrate was concentrated and purified by combi-flash (0-30% of EtOAc in PE) to give pure 4555 (6 mg, 15%, 4555) as a solid.

[0940] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.34 (s, 3H), 3.17-3.07 (m, 1H), 2.10-2.01 (m, 1H), 2.00-1.91 (m, 2H), 1.88-1.75 (m, 4H), 1.71-1.57 (m, 6H), 1.52-1.43 (m, 4H), 1.42-1.32 (m, 5H), 1.31-1.18 (m, 7H), 1.17-1.06 (m, 5H), 1.05-0.96 (m, 2H), 0.95-0.87 (m, 4H), 0.84 (s, 3H), 0.73-0.63 (m, 4H).

[0941] LCMS Rt=1.321 min in 2.0 min chromatography, 30-90AB_E, purity 100%.

[0942] MS 50-100_1_4min.m, MS ESI calcd. for C.sub.31H.sub.51F.sub.3O.sub.3Na [M+Na].sup.+ 551, found 551.

##STR00217##

Synthesis of ST-200-096-012A/B

[0943] ##STR00218##

[0944] To a solution of M-1-12_3 (500 mg, 0.747 mmol) in MeOH (40 mL) was added NiCl.sub.2 (19.2 mg, 0.149 mmol) and Mg powder (715 mg, 29.8 mmol) at 65° C. in one portion. The mixture was stirred at 65° C. for 10 minutes. Another Mg powder (355 mg, 14.9 mmol) was added in one portion. The mixture was stirred at 65° C. for 10 minutes again, quenched with HCl (200 mL, 1N) and extracted with EtOAc (3×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by combi-flash (0-15% of EtOAc in PE) to give ST-200-096-012A (57 mg, 14%, Peak 1) and ST-200-096-012B (26 mg, 6.6%, Peak 2) as a solid. 4555

[0945] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.34 (s, 3H), 3.17-3.07 (m, 1H), 2.09-1.91 (m, 3H), 1.88-1.76 (m, 4H), 1.70-1.61 (m, 5H), 1.56-1.44 (m, 6H), 1.43-1.19 (m, 11H), 1.17-0.95 (m, 7H), 0.95-0.85 (m, 4H), 0.84 (s, 3H), 0.72-0.61 (m, 4H).

[0946] LCMS Rt=1.269 min in 2.0 min chromatography, 30-90AB_E, purity 100%.

[0947] MS 50-100_1_4min.m, MS ESI calcd. for C.sub.31H.sub.51F.sub.3O.sub.3Na [M+Na].sup.+ 551, found 551.

[0948] 4585

[0949] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.39-3.34 (m, 1H), 3.31 (s, 3H), 2.11-2.02 (m, 2H), 1.98-1.91 (m, 1H), 1.87-1.75 (m, 4H), 1.73-1.60 (m, 6H), 1.56-1.33 (m, 10H), 1.32-1.05 (m, 10H), 1.04-0.93 (m, 3H), 0.93-0.86 (m, 4H), 0.84 (s, 3H), 0.72-0.63 (m, 4H).

[0950] LCMS Rt=1.257 min in 2.0 min chromatography, 30-90AB_E, purity 100%.

[0951] MS 50-100_1_4min.m, MS ESI calcd. for C.sub.31H.sub.51F.sub.3O.sub.3Na [M+Na].sup.+ 551, found 551.

Example 46: Synthesis of 4656 and 4657

[0952] ##STR00219## ##STR00220##

[0953] The synthesis of ST-200-CF3_6C can be found in Example 5.

Synthesis of M-1-15_6B

[0954] ##STR00221##

[0955] Tert-butyllithium (44.3 mL, 12.9 mmol, 1.3 M in n-hexane) was added to a solution of chloro(methoxymethyl)triphenylphosphorane (21.9 g, 64 mmol) in THF (100 mL) at 0° C. After stirring for 1 hour at 0° C., M-1-15_6A (5 g, 32.0 mmol) in THF (30 mL) was added at 0° C. and the reaction mixture was stirred at 15° C. for 12 hours. The reaction mixture was quenched with water (60 mL) and extracted with EtOAc (3×150 mL). The combined organic layers were dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the crude. The crude was purified by column chromatography with PE/EA=20/1-3/1 to give M-1-15_6B (5.55 g, 94%) as an oil.

[0956] .sup.1HNMR (400 MHz, CDCl.sub.3) δ 5.78 (s, 1H), 3.97-3.91 (m, 4H), 3.53 (s, 3H), 2.31 (t, J=6.4 Hz, 2H), 2.1-2.06 (m, 2H), 1.68-1.59 (m, 4H).

Synthesis of M-1-15_6C

[0957] ##STR00222##

[0958] Pd/C (1 g) under N.sub.2 at 10° C. was added to solution of M-1-15_6B (5.55 g, 30.1 mmol) in methanol (60 mL). The suspension was degassed under vacuum and purged with H.sub.2 three times. The mixture was stirred under H.sub.2 (50 psi) at 25° C. for 16 hours to give a suspension. The reaction mixture was filtered through a pad of Celite (2 cm) and the filter cake was washed with methanol (3×20 mL). The filtrate was concentrated to give M-1-15_6C (4.95 g, 88%) as an oil.

[0959] .sup.1HNMR (400 MHz, CDCl.sub.3) δ=3.97-3.88 (m, 4H), 3.32 (s, 3H), 3.20 (d, J=6.5 Hz, 2H), 1.75 (br d, J=9.3 Hz, 4H), 1.68-1.47 (m, 3H), 1.31-1.17 (m, 2H).

Synthesis of M-1-15_6

[0960] ##STR00223##

[0961] HCl (13.0 mL, 5 M) was added to a solution of M-1-156C (2 g, 10.7 mmol) in THF (20 mL). The reaction mixture was stirred at 10° C. for 48 hours, then 25° C. for 2 hours. The reaction mixture was concentrated to give a residue, which was basified to pH˜10 with NaOH (2 M) and extracted with EtOAc (2×20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give M-1-15_6 (1.25 g, 82%) as a liquid.

[0962] .sup.1HNMR (400 MHz, CDCl.sub.3) δ 3.35 (s, 3H), 3.30-3.25 (m, 2H), 2.44-2.28 (m, 4H), 2.15-2.06 (m, 2H), 2.05-1.95 (m, 1H), 1.50-1.37 (m, 2H).

Synthesis of M-1-15_7

[0963] ##STR00224##

[0964] A solution of t-BuOK (787 mg, 7.02 mmol) in THF (2.5 mL) was slowly added to a suspension of Me.sub.3IS (930 mg, 4.56 mmol) in THF (5 mL) was added under N2 at 15° C. After stirring at 15° C. for 30 min, then M-1-15_6 (0.5 g, 3.51 mmol) in 2.5 ml of THF was added drop wise to the mixture at 0° C. After the addition, the mixture was stirred at 15° C. for 16 hours, quenched with sat. NH.sub.4Cl (10 mL) and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give M-1-15_7 (410 mg, crude) as a liquid.

[0965] .sup.1HNMR (400 MHz, CDCl.sub.3) δ 3.34 (s, 3H), 3.30-3.25 (m, 2H), 2.65-2.55 (m, 2H), 1.95-1.80 (m, 4H), 1.75-1.60 (m, 1H), 1.40-1.15 (m, 4H).

Synthesis of M-1-15_8

[0966] ##STR00225##

[0967] A solution of n-BuLi (0.472 mL, 2.5 M in hexane, 1.18 mmol) was added to THF (0.5 mL). A solution of ST-200-CF3_6C (250 mg, 0.474 mmol) in THF (2.5 mL) was added to the mixture at −70° C. The mixture was stirred at −70° C. for 1 hour. M-1-15_7 (111 mg, 0.711 mmol) was added at −70° C. After stirring at −70° C. for another 1 hour, the mixture was warmed to 15° C. and stirred for 16 hrs. The reaction mixture was quenched with NH.sub.4Cl (50 mL, sat. aq) and extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give M-1-15_8 (250 mg, crude) as a solid, which was used for next step directly.

Synthesis of M-1-15A & M-1-15B

[0968] ##STR00226##

[0969] NiCl.sub.2 (9.48 mg, 0.0732 mmol) and Mg powder (350 mg, 14.6 mmol) were added in one portion to a solution of M-1-15_8 (350 mg, 0.513 mmol) in MeOH (30 mL) at 65° C. The mixture was stirred at 65° C. for 10 minutes. Then another portion of Mg powder (175 mg, 7.32 mmol) was added at 65° C. After stirring at 65° C. for another 10 minutes, the mixture was quenched with HCl (50 mL, 2N) until the reaction became clear and extracted with EtOAc (3×20 mL). The combined organic layer was washed with sat. NH.sub.4Cl (50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by silica gel chromatography (0-15% of EtOAc in PE) to give M-1-15A (22 mg, 11%, 4656,) and M-1-15B (54 mg, 27%, 4657) as a solid.

[0970] NMA-1-15A (4656)

[0971] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.27 (s, 3H), 3.24-3.19 (m, 2H), 2.10-2.02 (m, 1H), 1.99-1.92 (m, 2H), 1.88-1.77 (m, 2H), 1.76-1.55 (m, 10H), 1.52-1.34 (m, 8H), 1.32-1.21 (m, 5H), 1.19-0.98 (m, 9H), 0.96-0.87 (m, 4H), 0.84 (s, 3H), 0.72-0.62 (m, 4H).

[0972] LCMS Rt=1.327 min in 2.0 min chromatography, 30-90AB_E, purity 100%.

[0973] MS 50-100_1_4min.m, MS ESI calcd. for C.sub.32H.sub.52F.sub.3O.sub.2 [M+H−H.sub.2O].sup.+ 525, found 525.

[0974] NMA-1-15B (4657)

[0975] .sup.1HNMR (400 MHz, CDCl.sub.3) δ 3.33 (s, 3H), 3.23-3.19 (m, 2H), 2.10-2.01 (m, 1H), 1.99-1.92 (m, 2H), 1.88-1.77 (m, 2H), 1.73-1.55 (m, 8H), 1.53-1.43 (m, 5H), 1.41-1.20 (m, 12H), 1.19-1.07 (m, 4H), 1.06-0.96 (m, 3H), 0.96-0.86 (m, 4H), 0.84 (s, 3H), 0.72-0.62 (m, 4H).

[0976] LCMS Rt=1.377 min in 2.0 min chromatography, 30-90AB_E, purity 100%.

[0977] MS 50-100_1_4min.m, MS ESI calcd. for C.sub.32H.sub.52F.sub.3O.sub.2 [M+H−H2O].sup.+ 525, found 525.

Example 47: Synthesis of 4799

[0978] ##STR00227##

Synthesis of M-2-4_1

[0979] ##STR00228##

[0980] A solution of n-BuLi (0.476 mL, 2.5 M in hexane, 1.19 mmol) was added to THF (0.5 mL). A solution of ST-200-CF.sub.3_4A (250 mg, 0.476 mmol) in THF (2.5 mL) was added at −70° C. After stirring at −70° C. for 1 hour, ST-200-43-4_2 (100 mg, 0.714 mmol) was added to the mixture at −70° C. The mixture was stirred at −70° C. for another 1 hour. The mixture was warmed to 15° C., stirred for 16 hrs, quenched with NH.sub.4Cl (50 mL, sat. aq) and extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give M-2-4_1 (250 mg, crude) as a solid, which was used for next step directly.

Synthesis of 4799

[0981] ##STR00229##

[0982] NiCl.sub.2 (9.71 mg, 0.075 mmol) and Mg powder (360 mg, 15.0 mmol) were added in one portion to a solution of M-2-4_1 (250 mg, 0.375 mmol) in MeOH (30 mL) at 65° C. The mixture was stirred at 65° C. for 10 minutes. Then another portion of Mg powder (180 mg, 7.5 mmol) was added at 65° C. After stirring at 65° C. for another 10 minutes, the mixture was quenched with HCl (50 mL, 2N) until the reaction became clear and extracted with EtOAc (3×20 mL). The combined organic layer was washed with sat. NH.sub.4Cl (50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by silica gel chromatography (0-15% of EtOAc in PE) to give 4799 (56 mg, 28%) as a solid.

[0983] .sup.1HNMR (400 MHz, CDCl.sub.3) δ 5.40-5.33 (m, 1H), 2.48 (s, 2H), 2.08-1.92 (m, 4H), 1.91-1.69 (m, 3H), 1.62-1.56 (m, 2H), 1.53-1.45 (m, 9H), 1.44-1.35 (m, 4H), 1.34-1.23 (m, 2H), 1.22-1.04 (m, 10H), 1.04-0.97 (m, 2H), 0.96-0.90 (m, 6H), 0.87 (s, 3H), 0.68 (s, 3H).

[0984] LCMS Rt=1.439 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. for C.sub.32H.sub.50F.sub.3O [M+H−H.sub.2O].sup.+ 507, found 507.

Example 48: Synthesis of 4805

[0985] ##STR00230##

[0986] The synthesis of ST-200-CF3_6C can be found in Example 5.

Synthesis of M-1-20_2

[0987] ##STR00231##

[0988] A solution of t-BuOK (902 mg, 8.04 mmol) in THF (4 mL) was slowly added to a suspension of C.sub.3H.sub.9IS (1.06 g, 5.22 mmol) in THF (5 mL) under N.sub.2 at 15° C. After stirring at 20° C. for 30 min, M-1-20_1 (500 mg, 4.02 mmol) in 1 ml of THF was added dropwise to the mixture at 0° C. After addition, the mixture was stirred at 20° C. for 16 hours, quenched with sat. NH.sub.4Cl (40 mL) and extracted with MTBE (3×20 mL). The combined organic phase was washed with brine (2×60 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated at 40° C. under reduced pressure to give M-1-20_2 (390 mg, crude) as a liquid.

[0989] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.63 (s, 2H), 1.74-1.53 (m, 6H), 1.41-1.27 (m, 2H), 0.37-0.26 (m, 4H).

Synthesis of M-1-20_3

[0990] ##STR00232##

[0991] A solution of n-BuLi (0.472 mL, 2.5 M in hexane, 1.18 mmol) was added to THF (0.5 mL). A solution of ST-200-CF3_6C (250 mg, 0.474 mmol) in THF (2.5 mL) was added at −70° C. After stirring at −70° C. for 1 h, M-1-20_2 (98.2 mg, 0.711 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 h, warmed to 15° C. for 16 hours, quenched with NH.sub.4Cl (50 mL, sat. aq) and extracted with EtOAc (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, and concentrated to give M-1-20_3 (250 mg, crude) as a solid, which was used for next step directly.

Synthesis of 4805

[0992] ##STR00233##

[0993] NiCl.sub.2 (9.71 mg, 0.075 mmol) and Mg powder (360 mg, 15.0 mmol) were added in one portion to a solution of M-1-20_3 (250 mg, 0.375 mmol) in MeOH (30 mL) at 65° C. The mixture was stirred at 65° C. for 10 minutes. Then another portion of Mg powder (180 mg, 7.5 mmol) was added at 65° C. After stirring at 65° C. for another 10 minutes, the mixture was quenched with HCl (50 mL, 2N) until the reaction became clear and extracted with EtOAc (3×20 mL). The combined organic layer was washed with sat. NH.sub.4Cl (50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by silica gel chromatography (0-15% of EtOAc in PE) to give 4805 (150 mg, 76%) as a solid.

[0994] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.11-1.94 (m, 3H), 1.89-1.60 (m, 9H), 1.54-1.35 (m, 9H), 1.34-1.19 (m, 6H), 1.18-1.06 (m, 5H), 1.05-0.88 (m, 8H), 0.85 (s, 3H), 0.74-0.62 (m, 4H), 0.33-0.15 (m, 4H).

[0995] LCMS Rt=1.414 min in 2.0 min chromatography, 30-90AB_E, purity 100%.

[0996] MS 80-100_1_4min.m, MS ESI calcd. for C.sub.32H.sub.51F.sub.3O.sub.2Na [M+Na].sup.+ 547, found 547.

Example 49: Synthesis of 4906

[0997] ##STR00234## ##STR00235##

Synthesis of M-1-17_1

[0998] ##STR00236##

[0999] A solution of n-butyllithium (64 mL, 160 mmol, 2.5 M in hexane) was added to a solution of diisopropylamine (17.6 g, 174 mmol) in THF (30 mL) at −70° C. The mixture was warmed to 0° C. and stirred at 0° C. for 30 minutes. The mixture was cooled to −70° C. and M-1-15_2 (20 g, 69.8 mmol) in THF (20 mL) was added. The mixture was stirred at −70° C. for 1 h. Ethyl iodide (43.5 g, 279 mmol) was added. The mixture was warmed to 15° C. and stirred at 15° C. for 5 hours. The mixture was quenched with Sat NH.sub.4Cl (30 mL). The mixture was extracted with EtOAc (2×30 mL). The combined organic phase was washed with brine (2×20 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give a crude M-1-17_1 (21, crude) as an oil.

Synthesis of M-1-17_2

[1000] ##STR00237##

[1001] LiAlH.sub.4 (5.05 g, 133 mmol) was added in five portions to a solution of M-1-17_1 (21 g, 66.7 mmol) in THF (100 mL) at 0° C. under N.sub.2. The mixture was stirred at 20° C. for 4 hours. To the mixture water (20 mL) was added at 0° C. HCl (100 mL, 1 mol/L) was added. The aqueous phase was extracted with EtOAc (50 mL×2). The combined organic phase was washed with saturated brine (2×30 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-20% of EtOAc in PE) to give M-1-17_2 (16.5 g, 91%) as an oil.

[1002] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.70-3.55 (m, 1H), 3.55-3.45 (m, 2H), 1.70-1.55 (m, 5H), 1.55-1.10 (m, 9H), 0.95-0.88 (m, 9H), 0.041 (s, 6H).

Synthesis of M-1-17_3

[1003] ##STR00238##

[1004] 1-methyl-1H-imidazole (7.44 g, 90.7 mmol) and TEA (12.2 g, 121 mmol) were added to a solution of M-1-17_2 (16.5 g, 60.5 mmol) in DCM (100 mL) at 15° C. TsCl (23.0 g, 121 mmol) was added to the solution. The reaction mixture was stirred at 15° C. for 2 hours. The mixture was washed with water (2×100 mL), brine (150 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give M-1-17_3 (24 g, crude) as an oil.

Synthesis of M-1-17_4

[1005] ##STR00239##

[1006] LiAlH.sub.4 (5.32 g, 140 mmol) was added in five portions to a solution of M-1-17_3 (24 g, 56.2 mmol) in THF (100 mL) at 0° C. under N.sub.2. The mixture was stirred at 20° C. for 4 hours. Water (20 mL) was added to the mixture at 0° C. HCl (100 mL, 1 mol/L) was added. The aqueous phase was extracted with EtOAc (2×50 mL). The combined organic phase was washed with saturated brine (2×30 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give M-1-17_4 (13 g, crude) as an oil.

[1007] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.60-3.50 (m, 1H), 1.95-1.70 (m, 3H), 1.70-1.01 (m, 12H), 1.01-0.68 (m, 10H), 0.043 (s, 6H).

Synthesis of M-1-17_5

[1008] ##STR00240##

[1009] p-TsOH (7.23 g, 38.9 mmol) was added to a solution M-1-17_4 (10 g, 38.9 mmol) in acetone (50 mL). The reaction mixture was stirred at 15° C. for 2 h. Water was added and was extracted with EtOAc (2×30 mL). The combined organics were washed with NaHCO.sub.3 (20 mL, 10%) and brine (30 mL) and dried over Na.sub.2SO.sub.4. The solvent was removed under reduced pressure. The residue was purified by flash column (0-20% of EtOAc in PE) to give M-1-17_5 (6 g, crude) as a an oil.

[1010] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.70-3.50 (m, 1H), 1.80-1.60 (m, 4H), 1.60-1.40 (m, 5H), 1.40-1.01 (m, 4H), 0.91-0.75 (m, 4H).

Synthesis of M-1-17_6

[1011] ##STR00241##

[1012] DMP (17.8 g, 42 mmol) was added to a solution of M-1-17_5 (3 g, 21 mmol) in DCM (20 mL). Next, H.sub.2O (7.55 mg, 0.42 mmol) was added to the solution. The reaction was stirred at 15° C. for 30 min. Aqueous saturated NaHCO.sub.3 (10 mL) solution, aqueous saturated Na.sub.2S.sub.2O.sub.3 (10 mL) solution were added to the reaction mixture. The mixture was extracted with DCM (2×20 mL). The combined organic layer was washed with aqueous saturated NaHCO.sub.3 (2×20 mL) solution and brine (20 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum and purified by flash column (0-10% of EtOAc in PE) to give M-1-17_6 (2.5 g, 85%) as an oil.

[1013] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.40-2.28 (m, 4H), 1.72-1.60 (m, 4H), 1.49-1.40 (m, 2H), 1.02 (s, 3H), 0.92-0.85 (m, 3H).

Synthesis of M-1-17_7

[1014] ##STR00242##

[1015] M-1-17_6 (1 g, 7.13 mmol) was added to a stirred solution of trimethyl sulfoxonium iodide (3.12 g, 14.2 mmol) and t-BuOK (1.75 g, 15.6 mmol) in THF (10 mL) at 0° C. The reaction mixture was stirred at 10° C. for 16 hours. The reaction mixture was poured into saturated aqueous NH.sub.4Cl (15 mL). The resulting mixture was extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give M-1-17_7 (360 mg, crude) as an oil.

Synthesis of M-1-17_8

[1016] ##STR00243##

[1017] A solution of n-BuLi (0.568 mL, 2.5 M in hexane, 1.42 mmol) was added to THF (0.5 mL). A solution of ST-200-CF3_6C (300 mg, 0.474 mmol) in THF (2.5 mL) was added at −70° C. After stirring at −70° C. for 1 h, M-1-17_7 (131 mg, 0.853 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 h, warmed to 15° C. and stirred for 16 hours. The reaction mixture was quenched with NH.sub.4Cl (10 mL, sat. aq) and extracted with EtOAc (2×20 mL). The organic layer was separated, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give M-1-15_8 (387 mg, crude) as an oil, which was used for the next step directly.

Synthesis of 4906

[1018] ##STR00244##

[1019] NiCl.sub.2 (14.6 mg, 0.113 moml) and Mg powder (550 mg, 22.9 mmol) were added in one portion to a solution of M-1-17_8 (387 mg, 568 μmol) in MeOH (40 mL) at 65° C. After stirring at 65° C. for 10 minutes, another batch of Mg powder (266 mg, 11.1 mmol) was added in one portion at 65° C. The mixture was stirred at 65° C. for another 10 minutes. The reaction mixture was cooled to 20° C. and quenched by HCl (30 mL, 2 M). The resulting mixture was extracted with EtOAc (3×70 mL). The combined organic layers were washed with saturated aqueous NH.sub.4Cl (70 mL), brine (70 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the crude. The crude was purified by column chromatography with PE/EtOAc=0/1-5/1. The solvent was removed to give 4906 (56 mg, 18%) as a solid.

[1020] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.12-2.00 (m, 1H), 2.00-1.93 (m, 1H), 1.90-1.80 (m, 1H), 1.78-1.60 (m, 2H), 1.59-1.50 (m, 7H), 1.49-1.33 (m, 12H), 1.32-1.15 (m, 9H), 1.14-1.0 (m, 7H), 0.99-0.90 (m, 3H), 0.89-0.80 (m, 8H), 0.75-0.70 (m, 1H), 0.70-0.60 (s, 3H).

[1021] LCMS Rt=1.550 min in 2.0 min chromatography, 30-90AB_E, purity 100%; special MS ESI calcd. for C.sub.33H.sub.56F.sub.3O.sub.2 [M+H].sup.+ 541, found 541.

Example 50: Synthesis of 5009

[1022] ##STR00245##

[1023] The synthesis of ST-200-CF3_6C can be found in Example 5.

Synthesis of M-1-21_2

[1024] ##STR00246##

[1025] A solution of t-BuOK (797 mg, 7.12 mmol) in THF (3 mL) was added slowly by stirring into a suspension of C.sub.3H.sub.9IS (942 mg, 4.62 mmol) in THF (5 mL) under N.sub.2 at 15° C. After stirring at 20° C. for 30 min, M-1-21_1 (500 mg, 3.56 mmol) in 2 ml of THF was added dropwise to the mixture at 0° C. After addition, the mixture was stirred at 20° C. for 16 hrs, quenched with sat. NH.sub.4Cl (40 mL) without monitor and extracted with MTBE (3×20 mL). The combined organic phase was washed with brine (2×60 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated at 40° C. under reduced pressure to give M-1-21_2 (360 mg, crude) as a liquid.

[1026] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.47-4.42 (m, 4H), 2.61 (s, 2H), 2.06-1.90 (m, 4H), 1.68-1.58 (m, 2H), 1.51-1.41 (m, 2H).

Synthesis of M-1-21_3

[1027] ##STR00247##

[1028] A solution of n-BuLi (0.472 mL, 2.5 M in hexane, 1.18 mmol) was added to THF (0.5 mL). A solution of ST-200-CF3_6C (250 mg, 0.474 mmol) in THF (2.5 mL) was added at −70° C. After stirring at −70° C. for 1 h, M-1-21_2 (109 mg, 0.711 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 h and then warmed to 15° C. for 16 hrs. The reaction mixture was quenched with NH.sub.4Cl (50 mL, sat. aq) and extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give M-1-21_3 (250 mg, crude) as a solid, which was used for next step directly.

Synthesis of 5009

[1029] ##STR00248##

[1030] NiCl.sub.2 (9.49 mg, 0.0733 mmol) and Mg powder (350 mg, 14.6 mmol) were added in one portion to a solution of M-1-21_3 (250 mg, 0.367 mmol) in MeOH (30 mL) at 65° C. The mixture was stirred at 65° C. for 10 minutes. Then another Mg powder (178 mg, 7.34 mmol) was added at 65° C. in one portion. After stirring at 65° C. for another 10 minutes, the mixture was quenched with HCl (50 mL, 2N) until the reaction became clear and extracted with EtOAc (3×20 mL). The combined organic layer was washed with sat. NH.sub.4Cl (50 mL), NaHCO.sub.3 (50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by silica gel chromatography (0-15% of EtOAc in PE) to give impure 5009 (200 mg,), which was further purified by combi-flash (0-10% of Acetone in DCM) then recrystallized to give 120 mg still impure product. DMAP (13.4 mg, 0.11 mmol) and BzCl (77.3 mg, 0.550 mmol) were added to a solution of impure 5009 (60 mg, 0.110 mmol) in Py (5 mL). The reaction mixture was stirred at 20° C. for 2 hrs. The reaction was quenched with sat. NH.sub.4Cl (30 mL) and extracted with MTBE (2×15 mL). The combined organic phase was washed with brine (40 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated, and purified by prep-TLC (PE:EtOAc=5:1) to give desired product (40 mg, 56%) as a solid. To a solution of above (40 mg, 0.062 mmol) in MeOH (3 mL), THF (1 mL) and H.sub.2O (1 mL) was added NaOH (49.5 mg, 1.24 mmol). After stirring at 50° C. for 1 h, the reaction mixture was quenched with water (5 mL) and extracted with EtOAc (2×3 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) to give 5009 (17 mg, 50%) as a solid.

[1031] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.73 (s, 2H), 3.31 (s, 2H), 2.10-1.90 (m, 3H), 1.88-1.72 (m, 4H), 1.71-1.58 (m, 5H), 1.56-1.41 (m, 7H), 1.40-1.31 (m, 5H), 1.30-1.15 (m, 7H), 1.14-0.93 (m, 5H), 0.92-0.86 (m, 4H), 0.85 (s, 3H), 0.70-0.60 (m, 4H).

[1032] LCMS Rt=1.282 min in 2.0 min chromatography, 30-90AB_E, purity 100%.

[1033] MS 50-100_1_4min.m, MS ESI calcd. for C.sub.32H.sub.50F.sub.3O.sub.2 [M+H−H.sub.2O].sup.+ 523, found 523.

Example 51: Synthesis of 5131

[1034] ##STR00249##

[1035] The synthesis of ST-200-CF3_6C can be found in Example 5.

Synthesis of M-1-22_2

[1036] ##STR00250##

[1037] A solution of FSO.sub.2CF.sub.2COOH (18.3 g, 103 mmol) in CH.sub.3CN (30 mL) was added dropwise over a period of 1 hour to a mixture of M-1-22_1 (10 g, 86 mmol) and Na2SO.sub.4 (6.1 g, 43 mmol) in CH.sub.3CN (120 mL) at 40 to 45° C. After addition, the solution was poured into water (200 mL). The aqueous phase was extracted with DCM (3×100 mL). The combined organic phase was washed with saturated brine (2×200 mL), dried over anhydrous Na2SO.sub.4, filtered, concentrated and purified by flash column (0-100% of EtOAc in PE) to give M-1-22_2 (6 g, crude) as an oil.

[1038] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.42-6.02 (m, 1H), 4.30-4.10 (m, 1H), 3.80-3.60 (m, 1H), 2.20-1.30 (m, 8H).

Synthesis of M-1-22_3

[1039] ##STR00251##

[1040] Silica gel (5 g) and PCC (15.5 g, 72.2 mmol) were added to a suspension of M-1-22_2 (6 g, 36.1 mmol) in DCM (100 mL) at 20° C. After stirring at 20° C. for 2 hours, the mixture was filtered and the filter cake was washed with DCM (100 mL). The combined filtrate was concentrated in vacuum and purified by flash column (0-100% of EtOAc in PE) to give crude product M-1-22_3 (3.5 g, 59%) as a oil.

[1041] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.52-6.10 (m, 1H), 4.65-4.55 (m, 1H), 2.70-2.55 (m, 2H), 2.40-2.25 (m, 2H), 2.23-2.11 (m, 2H), 2.10-1.98 (m, 2H).

Synthesis of M-1-22_4

[1042] ##STR00252##

[1043] M-1-22_3 (3.1 g, 18.8 mmol) was added to a stirred solution of trimethylsulfoxonium iodide (4.97 g, 24.4 mmol) and t-BuOK (4.21 g, 37.6 mmol) in THF (60 mL) at 0° C. After stirring at 20° C. for 16 hours, the reaction mixture was poured into saturated aqueous NH.sub.4Cl (90 mL) and extracted with EtOAc (3×120 mL). The combined organic layers were washed with brine (120 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give M-1-22_4 (1.9 g, crude) as an oil, which was purified by combi-flash (0-10% of EtOAc in PE) to give pure M-1-22_4 (300 mg, 23%) as an oil.

[1044] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.50-6.02 (m, 1H), 4.46-4.35 (m, 0.6H), 4.33-4.23 (m, 0.4H), 2.65 (s, 2H), 2.07-1.86 (m, 5H), 1.74-1.58 (m, 2H), 1.47-1.36 (m, 1H).

Synthesis of M-1-22_5

[1045] ##STR00253##

[1046] A solution of n-BuLi (378 μL, 2.5 M in hexane, 0.947 mmol) was added to THF (0.5 mL). A solution of ST-200-CF3_6C (200 mg, 0.379 mmol) in THF (2 mL) was added at −70° C. The mixture was stirred at −70° C. for 1 h. M-1-22_4 (101 mg, 0.568 μmol) was added. After stirring at −70° C. for 1 h and 15° C. for 16 hours, the reaction mixture was quenched with sat. NH.sub.4Cl (10 mL) and extracted with EtOAc (2×5 mL). The organic layer was separated, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give M-1-22_5 (200 mg, crude) as a solid, which was used for the next step directly.

Synthesis of 5131

[1047] ##STR00254##

[1048] NiCl.sub.2 (7.13 mg, 0.0566 mmol) and Mg powder (270 mg, 11.3 mmol) were added in one portion to a solution of M-1-22_5 (200 mg, 0.283 mmol) in MeOH (30 mL) at 65° C. After stirring at 65° C. for 10 minutes, another batch of Mg powder (135 mg, 5.66 mmol) was added at 65° C. in one portion. The mixture was stirred at 65° C. for another 10 minutes, cooled to 20° C., quenched by HCl (20 mL, 2 M) and extracted with EtOAc (3×15 mL). The combined organic layers were washed with saturated aqueous NH.sub.4Cl (50 mL), brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give a crude, which was purified by combi-flash (0-15% of EtOAc in PE) to give 5131 (2 mg, 1.25%, 5131) as a solid.

[1049] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.45-6.02 (m, 1H), 4.12-4.00 (m, 1H), 2.11-2.02 (m, 1H), 1.97-1.92 (m, 2H), 1.85-1.77 (m, 6H), 1.71-1.62 (m, 5H), 1.51-1.43 (m, 4H), 1.41-1.34 (m, 5H), 1.33-1.25 (m, 4H), 1.24-1.19 (m, 2H), 1.16-0.99 (m, 7H), 0.93-0.87 (m, 4H), 0.84 (s, 3H), 0.68-0.63 (m, 4H).

[1050] LCMS Rt=5.826 min in 10.0 min chromatography, 50-100AB_E, purity 96.3%.

[1051] MS 50-100_1_4min.m, MS ESI calcd. for C.sub.31H.sub.48F.sub.5O.sub.2 [M+H−H.sub.2O].sup.+ 547, found 547.

Example 52: Synthesis of 5294

[1052] ##STR00255## ##STR00256##

[1053] The synthesis of M-2-13_6:

##STR00257## ##STR00258##

[1054] To a solution of M-2-131 (50 g, 165 mmol) in THF (500 mL) was added Pd/C (5 g, 10%) and pyridine (2.5 mL). Then the solution was hydrogenated under H.sub.2 balloon at 25° C. for 16 hrs. The mixture was filtered through a pad of celite and the filtrate was concentrated. The residue was dissolved in CH.sub.2Cl.sub.2 (500 mL), washed with aq HCl (100 mL, 1M), brine (300 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuum to afford M-2-13_2 (63 g, crude) as an oil.

[1055] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.08-3.95 (m, 1H), 2.82-2.72 (m, 1H), 2.71-2.58 (m, 2H), 2.52-2.31 (m, 1H), 2.31-2.21 (m, 1H), 2.21-2.03 (m, 4H), 2.02-1.87 (m, 2H), 1.70-1.60 (m, 3H), 1.58-1.45 (m, 3H), 1.43-1.22 (m, 4H), 1.14 (s, 3H), 0.88 (s, 3H).

[1056] To a suspension of M-2-13_2 (43 g, 141 mmol) in MeOH (200 mL) was added 4-methylbenzenesulfonic acid (2.42 g, 14.1 mmol) at 25° C. under N.sub.2. The mixture was stirred at 60° C. for 16 hrs. The reaction mixture was quenched with TEA (2 mL) and concentrated in vacuum to give M-2-13_3 (50 g, crude) as an oil, which was used directly for next step without further purification.

[1057] To a suspension of EtPPh.sub.3Br (158 g, 426 mmol) in THF (300 mL) was added t-BuOK (47.8 g, 426 mmol) at 25° C. under N.sub.2. The mixture was stirred at 60° C. for 30 mins. To the mixture was added M-2-13_3 (50 g, 142 mmol) in THF (300 mL) at 60° C. The mixture was stirred at 60° C. for 16 hrs. The mixture was added sat. NH.sub.4Cl solution (200 mL) and extracted with EtOAc (2×200 mL). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give crude product (200 g), which was used directly in next step without further purification. To a solution of the 200 g crude product in THF (500 mL) was added HCl (137.0 mL, 2 M in THF) at 25° C. The reaction was stirred at 25° C. for 1 h. The reaction was quenched with sat. NaHCO.sub.3 solution (200 mL) and extracted with EtOAc (2×200 mL). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give crude product (220 g). The crude product was purified by a silica gel column (PE/EtOAc=10/1-6/1) to give M-2-13_3A (43 g, impure), which was triturated with (PE/EtOAc=1/1) to give M-2-13_3A (18 g, pure, 42%) as a solid.

[1058] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.19-5.08 (m, 1H), 4.06-3.96 (m, 1H), 2.80-2.57 (m, 4H), 2.44-2.33 (m, 1H), 2.28-2.14 (m, 2H), 2.03-1.96 (m, 1H), 1.94-1.76 (m, 3H), 1.68-1.64 (m, 4H), 1.61-1.50 (m, 3H), 1.44-1.19 (m, 5H), 1.14 (s, 3H), 1.03-1.00 (m, 1H), 0.90 (s, 3H). To a solution of M-2-13_3A (8.7 g, 27.4 mmol) in THF (100 mL) was added TBAF (2.05 mL, 2.05 mmol, 1M in THF) and TMSCF.sub.3 (7.79 g, 54.8 mmol) under N.sub.2 at 10° C. The mixture was stirred at 10° C. for 1 h. To the mixture was added TBAF solution (82.1 mL, 82.1 mmol, 1M in THF). The mixture was stirred at 25° C. for another 1 h. The mixture was concentrated in vacuum. The residue was dissolved in EtOAc (100 mL), washed with water (2×100 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum to afford crude product (10 g), which was combined with the batch of 6.9 g of crude product (prepared from 5.8 g of M-2-13_3A) and purified by a silica gel column (PE/EtOAc=8/1-3/1) to give M-2-13_4 (1.1 g, 6%) as colorless oil and M-2-13_4A (9.3 g, 53%) as a solid

[1059] M-2-13_4:

[1060] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.17-5.08 (m, 1H), 4.01-3.89 (m, 1H), 2.62-2.56 (m, 1H), 2.44-2.32 (m, 2H), 2.28-2.15 (m, 1H), 2.00-1.88 (m, 3H), 1.87-1.73 (m, 3H), 1.69-1.59 (m, 6H), 1.55-1.25 (m, 6H), 1.23-1.13 (m, 2H), 1.09 (s, 3H), 0.95-0.89 (m, 1H), 0.87 (s, 3H).

[1061] M-2-13_4A:

[1062] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.18-5.07 (m, 1H), 4.02-3.88 (m, 1H), 2.59 (dd, J=11.8 Hz, J=5.0 Hz, 1H), 2.46-2.30 (m, 2H), 2.29-2.13 (m, 1H), 2.02 (s, 1H), 1.99-1.73 (m, 5H), 1.70-1.61 (m, 4H), 1.55-1.26 (m, 8H), 1.23-1.13 (m, 2H), 1.09 (s, 3H), 0.94 (d, J=6.4 Hz, 1H), 0.87 (s, 3H).

[1063] To a solution of M-2-13_4 (1.1 g, 2.84 mmol) in THF (30 mL) was added Borane-tetrahydrofuran complex (11.3 mL, 11.3 mmol, 1 M in THF) at 25° C. under N.sub.2. The solution was stirred at 25° C. for 1 h. After cooling to 0° C., a solution of EtOH (30 mL) and NaOH (5.67 mL, 5M in H.sub.2O, 28.4 mmol) was added very slowly. After addition, H.sub.2O.sub.2 (2.84 mL, 28.4 mmol, 30% in water) was added slowly and the inner temperature was maintained below 10° C. The mixture was stirred at 25° C. under N.sub.2 for 1 h. Water (100 mL) was added to the solution and extracted with EtOAc (2×50 mL). The combined organic layer was washed sat. Na.sub.2S.sub.2O.sub.3 solution (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give M-2-13_5 (1 g, crude) as colorless oil, which was directly used for next step.

[1064] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.95-3.78 (m, 1H), 3.72-3.65 (m, 1H), 2.71-2.62 (m, 1H), 2.47-2.40 (m, 1H), 2.19-2.11 (m, 1H), 2.10-2.05 (m, 1H), 2.04-2.01 (m, 1H), 2.00-1.84 (m, 3H), 1.77-1.62 (m, 5H), 1.50-1.27 (m, 10H), 1.21-1.14 (m, 3H), 1.11 (s, 3H), 0.98-0.93 (m, 1H), 0.67 (s, 3H).

[1065] To a solution of M-2-13_5 (1 g, 2.47 mmol) in DCM (30 mL) was added silica gel (3 g) and PCC (2.65 g, 12.3 mmol) at 25° C. The reaction was stirred at 25° C. for 16 hrs. The reaction mixture was filtered and the filtrate was concentrated in vacuum to give crude product which was purified by a silica gel column (PE/EtOAc=5/1) to give M-2-13_6 (720 mg, 73%) as a solid. The solid was triturated with MeCN (10 mL) to afford M-2-13_6 (12 mg) as a solid and organic layer was concentrated in vacuum to afford M-2-13_6 (700 mg, 99%) as a solid which was used directly for next step.

[1066] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.75 (t, J=9.0 Hz, 1H), 2.63-2.43 (m, 3H), 2.33-2.17 (m, 2H), 2.10 (s, 3H), 2.02-1.75 (m, 6H), 1.74-1.70 (m, 1H), 1.68 (s, 1H), 1.66-1.58 (m, 1H), 1.53-1.22 (m, 7H), 1.21 (s, 3H), 0.58 (s, 3H).

[1067] LCMS Rt=0.911 min in 2 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.22H.sub.32F.sub.3O.sub.3 [M+H].sup.+ 401, found 401.

Synthesis of M-2-13_7

[1068] ##STR00259##

[1069] t-BuOK (835 mg, 7.45 mmol) was added to a suspension of MePPh.sub.3Br (2.66 g, 7.45 mmol) in THF (30 mL) at 25° C. under N.sub.2. After stirring at 50° C. for 30 mins, the mixture was added to a solution of M-2-136 (600 mg, 1.49 mmol) in THF (30 mL) at 25° C. The mixture was stirred at 25° C. for 16 hrs. The mixture was quenched with sat. NH.sub.4Cl solution (100 mL) and extracted with EtOAc (2×100 mL). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to afford M-2-13_7 (4 g, crude) as an oil, which was purified by combi-flash (EtOAc in PE, 10%) to afford M-2-13_7 (540 mg, 12%) as a solid.

[1070] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.89 (s, 1H), 4.71 (s, 1H), 2.53-2.20 (m, 6H), 2.02-1.73 (m, 7H), 1.72-1.60 (m, 5H), 1.56-1.23 (m, 7H), 1.21 (s, 3H), 0.53 (s, 3H).

Synthesis of M-2-13_8

[1071] ##STR00260##

[1072] 9-BBN dimer (988 mg, 4.05 mmol) was added to a solution of M-2-13_7 (540 mg, 1.35 mmol) in THF (20 mL) at 0° C. under N.sub.2. The solution was stirred at 25° C. for 16 hrs. After cooling to 0° C., a solution of EtOH (10 mL) and NaOH (2.70 mL, 5M in H.sub.2O, 13.5 mmol) were added very slowly. After addition, H.sub.2O.sub.2 (1.35 mL, 13.5 mmol, 30% in water) was added slowly and the inner temperature was maintained below 10° C. The mixture was stirred at 50° C. under N.sub.2 for 1 h. The mixture was cooled to 30° C., diluted with water (30 mL) and extracted with EtOAc (2×50 mL). The combined organic layer was washed with sat. Na.sub.2S.sub.2O.sub.3 (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give M-2-13_8 (2 g, crude) as an oil, which was directly used in next step without further purification.

Synthesis of M-2-13_9

[1073] ##STR00261##

[1074] TsCl (4.55 g, 23.9 mmol) was added to a solution of M-2-138 (2 g, crude) in DCM/TEA (16 mL/2.3 mL) at 25° C. The mixture was stirred at 40° C. for 2 hrs. The reaction was quenched with water (20 mL) and extracted with DCM (2×30 mL). The combined organic layer was washed with brine (2×50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to afford crude product, which was purified by combi-flash column (EtOAc in PE, 12%˜15%) to afford M-2-13_9 (580 mg, 21%) as an oil.

[1075] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.78 (d, J=8.4 Hz, 2H), 7.34 (d, J=8.0 Hz, 2H), 4.15-4.13 (m, 1H), 3.98-3.94 (m, 1H), 3.81-3.76 (m, 1H), 2.45 (s, 3H), 2.12-2.06 (m, 1H), 2.02-1.89 (m, 3H), 1.85-1.63 (m, 8H), 1.53-1.29 (m, 6H), 1.22 (s, 3H), 1.19-1.02 (m, 6H), 0.99 (d, J=6.8 Hz, 3H), 0.85 (s, 3H).

Synthesis of M-2-13_10A

[1076] ##STR00262##

[1077] KI (838 mg, 5.05 mmol) was added to a solution of M-2-139 (580 mg, 1.01 mmol) in DMF (6 mL) at 25° C. under N.sub.2. After stirring at 50° C. for 2 hours under N.sub.2, the reaction mixture was treated with PhSO.sub.2Na (820 mg, 5.00 mmol) and stirred at 50° C. for 16 hrs. The reaction mixture was cooled to 25° C. and treated with water (50 mL). The aqueous phase was extracted with EtOAc (2×50 mL). The combined organic phase was washed with saturated brine (2×100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated and purified by a silica gel column (PE/EtOAc=8/1-5/1) to afford M-2-13_10A (460 mg, 85%) as a solid.

[1078] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.93-7.88 (m, 2H), 7.68-7.62 (m, 1H), 7.60-7.53 (m, 2H), 4.15-4.12 (m, 1H), 3.15-3.09 (m, 1H), 2.87-2.80 (m, 1H), 2.13-2.07 (m, 2H), 2.03-1.89 (m, 3H), 1.83-1.64 (m, 6H), 1.47-1.27 (m, 5H), 1.23-1.18 (m, 7H), 1.18-0.95 (m, 7H), 0.87 (s, 3H).

Synthesis of M-2-13_10

[1079] ##STR00263##

[1080] Silica gel (600 mg) and PCC (571 mg, 2.65 mmol) were added to a solution of M-2-1310A (480 mg, 0.884 mmol) in DCM (15 mL) at 25° C. After stirring at 25° C. for 16 hrs, the reaction mixture was filtered and the filtrate was concentrated in vacuum to afford crude product, which was purified by combi-flash column (EtOAc in PE, 15%) to afford M-2-13_10 (280 mg, 59%) as a solid.

[1081] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.93-7.88 (m, 2H), 7.69-7.62 (m, 1H), 7.60-7.54 (m, 2H), 3.13-3.08 (m, 1H), 2.92-2.82 (m, 1H), 2.61-2.54 (m, 1H), 2.32-2.28 (m, 1H), 2.23-2.16 (m, 1H), 2.14-2.01 (m, 1H), 1.98-1.84 (m, 4H), 1.78-1.65 (m, 5H), 1.53-1.32 (m, 6H), 1.27-1.24 (m, 4H), 1.19 (s, 3H), 1.14 (d, 7=6.4 Hz, 3H), 0.62 (s, 3H).

Synthesis of M-2-13_11

[1082] ##STR00264##

[1083] DIPA (233 mg, 2.31 mmol) and n-BuLi (0.852 mL, 2.5 M in hexane, 2.13 mmol) were added to THF (0.5 mL) at −78° C. under N.sub.2. The mixture was warmed to 0° C. After re-cooling to −78° C., a solution of M-2-13_10 (330 mg, 0.610 mmol) in THF (2.5 mL) was added. The mixture was stirred at −78° C. for 1 h and treated with 6,6-dimethyl-1-oxaspiro[2.5]octane (128 mg, 0.915 mmol). The mixture was stirred at −78° C. for another 1 h, warmed to 25° C. and stirred at 25° C. for 16 hrs. The reaction mixture was quenched with NH.sub.4Cl (50 mL, sat. aq) and extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give crude product (380 mg) as an oil, which was used directly for next step without further purification.

Synthesis of 5294

[1084] ##STR00265##

[1085] Mg powder (1.07 g, 44.6 mmol) was added to a solution of M-002-013_11 (380 mg, 0.558 mmol) in 40 mL of dry methanol under N.sub.2 at 60° C. The reaction mixture was quenched with HCl (50 mL, 1 M in H.sub.2O) dropwise at 10° C. until solid was dissolved. After extracting with DCM (2×100 mL), the organic layer was washed with sat. NaHCO.sub.3 (50 mL), brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column eluted with PE/EtOAc=10/1-8/1 to give 5294 (35 mg, impure) as an oil. 5294 (35 mg, impure) was purified by a silica gel column (PE/EtOAc=10/1-8/1) to give 5294 (20 mg, impure) as an oil. 5294 (20 mg, 0.037 mmol) was purified by a silica gel column (DCM/acetone=40/1) for the third time to give 5294 (18 mg, impure) as an oil. The impure 5294 (18 mg, impure) was purified by prep. HPLC (column: Xtimate C18 150*25 mm*5 um, gradient: 90-100% B (A=0.1% TFA-ACN, B=acetonitrile), flow rate: 30 mL/min) to give 5294 (1.9 mg, 11%) as a solid. 1H NMR (400 MHz, CDCl3) δ 2.59-2.43 (m, 2H), 2.37-2.16 (m, 2H), 2.05-1.84 (m, 3H), 1.76-1.65 (m, 3H), 1.51-1.38 (m, 14H), 1.26-1.20 (m, 14H), 0.93 (s, 3H), 0.91-0.86 (m, 7H), 0.63 (s, 3H).

[1086] LCMS Rt=1.284 min in 2 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.32H.sub.50F.sub.3O.sub.2 [M+H−H.sub.2O].sup.+ 523, found 523.

Example 53. Biological Data

[1087] Experiments were conducted as described in Example 2. The results are shown in Table 2-59.

TABLE-US-00003 TABLE 2-59 Avg Avg Avg EC50 Avg Emax EC50 2B Emax Compound 2A (nM) 2A (%) (nM) 2B (%) [00266] 1839 295.4 493.3 148.2 568.5 [00267] 1 144.9 887.0 70.9 441.9 [00268] 2 162.7 708.9 147.1 605.2 [00269] 2A 157.9 652.6 339.1 1239.9 [00270] 2B 130.7 624.6 142.8 965.3 [00271] 10 125.4 631.7 101.6 362.3 [00272] 1-A 109.4 274.6 81.2 250.1 [00273] 1-B 31.6 262.5 33.6 284.6 [00274] 7 299.5 516.3 286.9 483.6 [00275] 9 316.6 180.6 234.0 271.4 [00276] 3 185.2 555.6 338.0 531.8 [00277] 1967 170.8 347.4 113.3 373.9 [00278] 7-B 346.8 355.7 305.8 352.1 [00279] 7-A 247.5 544.2 187.3 431.0 [00280] 8 446.1 401.7 372.4 321.7 [00281] 4 201.2 663.0 212.5 462.9 [00282] 5 181.8 619.0 260.4 454.8 [00283] 6 247.7 136.6 733.2 162.7 [00284] 11 116.9 151.2 104.3 157.4 [00285] 2080 165.1 77.2 186.9 146.9 [00286] 2081 128.8 125.8 174.7 214.9 [00287] 2184 65.8 233.7 80.0 257.5 [00288] 2285 763.6 204.7 494.3 219.6 [00289] 2392 104.4 230.1 47.5 166.2 [00290] 2499 768.1 179.8 750.7 237.6 [00291] 2500 63.3 302.6 91.1 300.1 [00292] 2602 288.6 162.6 279.0 366.6 [00293] 2706 >10000 96.8 124.0 139.3 [00294] 2707 119.0 112.8 102.7 259.0 [00295] E-2817 5065.7 29.9 5373.5 20.4 [00296] 2918 298.0 532.0 338.9 506.1 [00297] 3035 61.0 323.4 75.7 611.3 [00298] 3149 69.7 292.1 60.7 471.7 [00299] 3266 147.5 82.6 377.4 134.7 [00300] 3382 67.6 309.2 125.8 460.6 [00301] 3495 96.7 107.2 168.4 145.9 [00302] 3496 35.11 319.0 47.6 369.3 [00303] 3507 165.0 165.5 190.7 241.1 [00304] 3634 422.6 220.6 404.5 378.7 [00305] 3788 1317.9 229.4 1121.4 410.2 [00306] 3877 >10000 42.4 512.1 71.1 [00307] 3983 88.1 175.9 248.0 300.4 [00308] 4023 515.9 322.1 405.7 418.3 [00309] 4155 441.7 163.5 611.9 198.1 [00310] 4156 >10000 10.6 >10000 20.1 [00311] 4258 1025.3 183.9 640.5 241.7 [00312] 4259 >10000 −13.3 >10000 24.3 [00313] 4360 >10000 38.1 >10000 14.6 [00314] 4475 461.3 363.5 265.7 354.6 [00315] 4476 >10000 −3.3 >10000 10.4 [00316] 4555 >10000 22.8 >10000 31.9 [00317] 4656 1601.0 134.4 2164.5 122.0 [00318] 4657 422.2 74.0 129.5 62.9 [00319] 4799 >10000 36.1 760.2 32.8 [00320] 480.5 356.1 175.3 362.3 129.4 [00321] 4906 >10000 8.8 >10000 14.8 [00322] 5009 >10000 27.5 >10000 14.5 [00323] 5131 33.6 52.9 >10000 39.0 [00324] 5294 >10000 32.4 >10000 13.9 [00325] 4585 214.6 72.2 166.3 123.0 [00326] 3886 53.4 106.0 50.1 164.1

Example 54: Synthesis of Compound 154

[1088] ##STR00327##

Step 1

[1089] To a solution of A154 (2 g, 5.01 mmol) (see WO2014/160480 for synthesis) 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 crude A254 (1.8 g) as a solid.

Step 2

[1090] To a solution of A254 (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.4Cl (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 crude A354 (1.6 g) as a solid.

Step 3

[1091] A mixture of A354 (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 A454 (0.9 g, 54%) as a solid.

Step 4

Step 4a: Generation of 4-Pyridylmagnesium Chloride Solution

[1092] To a suspension of 4-bromopyridine hydrochloride (1 g, 5.14 mmol) in THF (4 mL) was added isopropylmagnesium chloride (5.1 mL, 2 M in THF, 10.2 mmol) at 0° C. The mixture was stirred at 15° C. for 1 h. The 4-pyridylmagnesium chloride solution (ca. 0.5 M in THF) was used directly.

Step 4b

[1093] To a solution of A454 (100 mg, 0.268 mmol) in THF (1 mL) was added freshly prepared 4-pyridylmagnesium chloride (5.36 mL, ca. 0.5 M in THF, 2.68 mmol) at 0° C. The mixture was stirred at 15° C. for 1 h. To the mixture was added NH.sub.4Cl (2 mL, 10% aq.). The mixture was extracted with EtOAc (10 mL). The organic layer was separated, purified by prep-TLC (DCM:MeOH=15:1), re-crystallized from MeCN (2 mL) and dried in vacuum to give Compound 154 (31 mg, 26%) as a solid.

[1094] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.57 (d, J=4.4 Hz, 2H), 7.28-7.26 (m, 2H), 5.34-5.26 (m, 1H), 4.71-4.58 (m, 1H), 2.47-2.37 (m, 1H), 2.03-1.90 (m, 4H), 1.85-1.61 (m, 5H), 1.56-1.46 (m, 8H), 1.39-1.04 (m, 9H), 1.04-0.85 (m, 9H), 0.70-0.62 (m, 3H).

[1095] LCMS Rt=0.806 min in 2.0 min chromatography, 30-90AB, MS ESI calcd. for C.sub.30H.sub.46NO.sub.2 [M+H].sup.+ 452, found 452.

Example 55. Synthesis of Compound 255

[1096] ##STR00328##

[1097] To a solution of 3-bromopyridine (423 mg, 2.68 mmol) in THF (10 mL) under N.sub.2 was added i-PrMgCl (1.34 mL, 2.68 mmol, 2M) dropwise at 15° C. The reaction was stirred at 15° C. for 30 min. A solution of A455 (100 mg, 0.268 mmol) was added. The reaction was stirred at 15° C. for 2 h. The reaction was quenched with saturated NH.sub.4Cl (20 mL), extracted with EtOAc (3×20 mL). The combined organic phase was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by column chromatography (MeOH in DCM gradient, 0%-10%) to afford crude product (50 mg), which was then recrystallized from MeCN (15 mL) to afford Compound 255 (8 mg, 7% yield) as a solid.

[1098] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.60-8.55 (m, 1H), 8.55-8.51 (m, 1H), 7.73-7.68 (m, 1H), 7.32-7.28 (m, 1H), 5.32-5.27 (m, 1H), 4.73-4.63 (m, 1H), 2.45-2.37 (m, 1H), 2.01-1.65 (m, 9H), 1.56-1.33 (m, 9H), 1.28-1.04 (m, 8H), 1.03-0.86 (m, 9H), 0.66 (s, 3H).

Example 56. Syntheses of Compounds 356 and 456

[1099] ##STR00329##

Step 1

[1100] To a solution of A456 (300 mg, 0.8 mmol) in THF (5 mL) was added phenylmagnesium bromide (2.01 mL, 1 M in ether, 2.01 mmol) dropwise at −60° C. The mixture was stirred at 25° C. for 1 h. The mixture was poured into water (50 mL) and extracted with EtOAc (2×50 mL). The combined organic layer was washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to afford Compound 356 (315 mg, crude) as a solid. 115 mg of Compound 356 was purified by prep-HPLC separation (column: Phenomenex Synergi C18 150*30 mm*4 um, gradient: 95% B (water (0.05% HCl)-ACN), flow rate: 25 mL/min) to give Compound 356 (31 mg) as a solid.

[1101] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.34-7.28 (m, 4H), 7.27-7.24 (m, 1H), 5.31-5.30 (m, 1H), 4.57-4.51 (m, 1H), 2.45-2.42 (m, 1H), 2.02-1.96 (m, 3H), 1.95-1.78 (m, 5H), 1.60-1.52 (m, 9H), 1.20-0.72 (m, 18H), 0.72-0.71 (m, 3H).

[1102] LCMS Rt=1.248 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.31H.sub.43 [M+H−2H.sub.2O].sup.+ 415, found 415.

Step 2

[1103] A mixture of Compound 356 (200 mg, 0.44 mmol) in DCM (3 mL) was added PCC (190 mg, 0.89 mmol) at 25° C. for 1 h. The solution was filtered and the filtered cake was washed with DCM (2×10 mL). The combined filtrate was concentrated in vacuum. The residue was purified by silica gel column eluted with (PE/EtOAc=10/1) to give B156 (150 mg, 72%) as a solid.

[1104] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.01-7.95 (m, 2H), 7.58-7.54 (m, 1H), 7.50-7.45 (m, 2H), 5.32-5.30 (m, 1H), 3.03-2.90 (m, 2H), 2.41-2.40 (m, 1H), 2.05-1.96 (m, 7H), 1.52-1.48 (m, 9H), 1.17-0.94 (m, 16H), 0.70 (s, 3H).

Step 3

[1105] To a solution of B156 (80 mg, 0.18 mmol) in THF (5 mL) was added dropwise MeLi (0.28 mL, 1.6 M in ether, 0.4 mmol) at −60° C. The mixture was stirred at 25° C. for 1 h. The mixture was poured into water (50 mL) and extracted with EtOAc (2×50 mL). The combined organic layer was washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by prep-HPLC separation (column: Phenomenex Synergi C18 150*30 mm*4 um, gradient: 65-95% B (water (0.05% HCl)-ACN), flow rate: 25 mL/min) to give Compound 456 (24 mg, 29%) as a solid.

[1106] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.42-7.40 (m, 2H), 7.32-7.28 (m, 2H), 7.20-7.17 (m, 1H), 5.29-5.28 (m, 1H), 2.43-2.40 (m, 1H), 1.98-1.93 (m, 4H), 1.75-1.50 (m, 16H), 1.48-0.88 (m, 18H), 0.67-0.65 (m, 3H).

[1107] LCMS Rt=1.289 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.32H.sub.45 [M+H−2H.sub.2O].sup.+ 429, found 429.

Example 57. Synthesis of Compound C0

[1108] ##STR00330## ##STR00331##

Step 1

[1109] To a mixture of MePPh.sub.3Br (1.28 kg, 3.6 mol) in THF (4.5 L) was added t-BuOK (404 g, 3.6 mol) at 15° C. under N.sub.2. The resulting mixture was stirred at 50° C. for 30 min. Pregnenolone (950 g, 2.9 mol) was added in portions below 65° C. The reaction mixture was stirred at 50° C. for 1 hour. The combined mixture was quenched with saturated NH.sub.4Cl aqueous (1 L) at 15° C. and the THF layer was separated. The aqueous layer was extracted with EtOAc (2×2 L). The combined organic phase was concentrated under vacuum to give a solid. The solid was further purified by trituration with MeOH/H.sub.2O (1:1, 15 L) at reflux to give C.sub.0-1 (940 g, 99%) as a solid.

[1110] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.32 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H), 3.58-3.46 (m, 1H), 2.36-2.16 (m, 2H), 2.08-1.94 (m, 2H), 1.92-1.62 (m, 9H), 1.61-1.39 (m, 6H), 1.29-1.03 (m, 4H), 1.01 (s, 3H), 0.99-0.91 (m, 1H), 0.59 (s, 3H).

Step 2

[1111] To a solution of C0-1 (800 g, 2.54 mol) in DCM (8 L) was added DMP (2.14 kg, 5.08 mol) in portions at 35° C. The reaction mixture was stirred at 35° C. for 20 mins. The reaction mixture was filtered. The filtered cake was washed with DCM (3 xl L). The combined organic phase was washed with saturated Na.sub.2S.sub.2O.sub.3/saturated NaHCO.sub.3 aqueous (3:1, 2×1.5 L), brine (1.5 L), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give C0-2 (794 g, crude) as a solid, which was used for next step directly.

Step 3

[1112] To a solution of BHT (1.97 kg, 8.94 mol) in toluene (1 L) was added AlMe.sub.3 (2.14 L, 2.0 M in toluene, 4.28 mol) drop-wise below 25° C. under N.sub.2 atmosphere. The resulting mixture was stirred at 25° C. for 1 hour. C0-2 (794 g, 2.16 mol) in DCM (3 L) was added at −70° C. The mixture was stirred at −70° C. for 1 hour. MeMgBr (862 mL, 3.0 M in diethyl ether, 2.59 mol) was added at −70° C. The reaction mixture was stirred at −70° C. for 10 min. The mixture was quenched by saturated critic acid (3 L), extracted with EtOAc (2×2 L). The combined organic phase was washed with brine (2 L), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a residue, which was triturated from MeCN (3 L) at 25° C. to give C0-3 (340 g, 43%) as a solid.

[1113] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.34-5.26 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H), 2.50-2.35 (m, 1H), 2.07-1.94 (m, 3H), 1.91-1.84 (m, 1H), 1.83-1.63 (m, 8H), 1.58-1.33 (m, 6H), 1.27-1.13 (m, 3H), 1.12 (s, 3H), 1.10-1.05 (m, 1H), 1.02 (s, 3H), 1.00-0.92 (m, 1H), 0.58 (s, 3H).

Step 4

[1114] To a mixture of C0-3 (149 g, 453 mmol) and 9-BBN dimer (127 g, 520 mmol) was added THF (1 L) at 15° C. under N.sub.2. The reaction mixture was stirred at 60° C. for 1 hour. The mixture was cooled to 15° C. EtOH (208 g, 4.53 mol) was added at 15° C. NaOH aqueous (906 mL, 5 M, 4.53 mol) was added drop-wise at 15° C. H.sub.2O.sub.2 (514 g, 30%, 4.53 mol) was added dropwise at 15° C. The obtained mixture was stirred at 60° C. for 1 hour. A solid was produced. The solid was washed with ethanol (200 mL) to give a solid, which was triturated with EtOH (2.3 L) at reflux and water (2.5 L) at 80° C. successively to give C0-4 (131 g, 84%) as a solid.

[1115] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.35-5.24 (m, 1H), 3.67-3.61 (m, 1H), 3.42-3.33 (m, 1H), 2.50-2.35 (m, 1H), 2.07-1.92 (m, 3H), 1.88-1.65 (m, 3H), 1.60-1.38 (m, 9H), 1.37-1.26 (m, 1H), 1.26-1.12 (m, 4H), 1.11 (s, 3H), 1.08 (s, 1H), 1.05 (d, J=6.8 Hz, 3H), 1.01 (s, 3H), 1.00-0.91 (m, 1H), 0.70 (s, 3H).

Step 5

[1116] To a solution of C0-4 (131 g, 378 mmol) in CHCl.sub.3 (600 mL) and pyridine (420 mL) was added TsCl (187 g, 982 mmol) at 15° C. The mixture was stirred at 15° C. for 2 hrs. The reaction mixture was concentrated under vacuum to remove most of CHCl.sub.3. Water (3 L) was added. A solid was produced and filtered. The solid was washed with water (6×4 L) and dissolved in DCM (3.5 L), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give C0-5 (177 g, 94%) as a solid.

[1117] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.78 (d, J=8.4 Hz, 2H), 7.34 (d, 7=8.0 Hz, 2H), 5.34-5.25 (m, 1H), 3.96 (dd, 7=3.2, 9.6 Hz, 1H), 3.79 (dd, J=6.4, 9.2 Hz, 1H), 2.45 (s, 3H), 2.50-2.35 (m, 1H), 2.02-1.88 (m, 3H), 1.81-1.61 (m, 4H), 1.58-1.33 (m, 8H), 1.24-1.12 (m, 4H), 1.11 (s, 3H), 1.09-1.01 (m, 2H), 1.00 (s, 3H), 0.98-0.86 (m, 3H), 0.64 (s, 3H).

Step 6

[1118] To a solution of C0-5 (177 g, 353 mmol) in DMF (1.8 L) was added KI (281 g, 1694 mmol) at 15° C. The mixture was stirred at 60° C. for 1 h. To the DMF mixture was added PhSO.sub.2Na (211 g, 1.06 mol). The mixture was stirred at 60° C. for 2 hrs. The reaction mixture was cooled to 25° C. The mixture was poured into water (20 L) and some solid was produced. The mixture was filtered. The filtered cake was washed with water (3×2 L) and dissolved in DCM (5 L). The solution was washed with water (2×1 L), brine (2×1 L), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum to give a crude product as a solid, which was re-crystallized from toluene (2.5 L) to give CO (121 g, 73%) as a solid. The re-crystallization filtrate was concentrated under vacuum to give a crude CO (20 g) as a solid.

[1119] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.91 (d, J=7.5 Hz, 2H), 7.69-7.61 (m, 1H), 7.61-7.53 (m, 2H), 5.31-5.24 (m, 1H), 3.14 (d, 7=14.0 Hz, 1H), 2.85 (dd, 7=9.2, 14.0 Hz, 1H), 2.50-2.35 (m, 1H), 2.16-2.03 (m, 1H), 2.01-1.88 (m, 3H), 1.80-1.64 (m, 3H), 1.56-1.34 (m, 7H), 1.20 (d, 7=6.8 Hz, 3H), 1.17-1.11 (m, 3H), 1.10 (s, 3H), 1.08-1.01 (m, 2H), 1.00 (s, 3H), 0.98-0.87 (m, 2H), 0.65 (s, 3H).

Example 58. Syntheses of Compound C3-1 and C3-2

[1120] ##STR00332##

Step 1

[1121] To a solution of t-BuOK (3.22 g, 28.7 mmol) in DMSO (30 mL) was added trimethylsulfonium iodide (6.42 g, 31.5 mmol) at 20° C. and stirred for 30 min under N.sub.2. A solution of C1 (5 g, 28.7 mmol) in DMSO (8 mL) was added and stirred for 16 h at 20° C. The reaction mixture was diluted with EtOAc (50 mL) and water (50 mL). The aqueous layer was back-extracted with EtOAc (50 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give crude product which was purified by a silica gel column (PE/EtOAc=3/1) to give C2 (1.9 g, 35%) as an oil.

[1122] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.55-7.52 (m, 2H), 7.43-7.35 (m, 3H), 3.43-3.40 (m, 1H), 2.95-2.92 (m, 1H).

Step 2

[1123] To THF (2 mL) under N.sub.2 at −70° C. was added n-BuLi (1.7 mL, 4.24 mmol). After that, a suspension of CO (500 mg, 1.06 mmol) in THF (5 mL) was added drop-wise to give a suspension. After stirring at −70° C. for 30 min, a solution of C2 (398 mg, 2.12 mmol) in THF (2 mL) was added. Then reaction was stirred at −70° C. for 10 min and then stirred at 20° C. for 16 hours. The reaction mixture was quenched with water (10 mL). The mixture was extracted with EtOAc (3×50 mL). The combined organic phases were washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give crude product which was purified by a silica gel column (PE/EtOAc=10/1) to give C3-1 (320 mg, 46%) as a solid and C.sub.3-2 (50 mg, impure). C3-1 was used directly for the next step. C.sub.3-2 was purified by HPLC separation (column: Phenomenex Synergi C18 150*30 mm*4 um, gradient: 82-95% B (A=0.05% HCl-ACN, B=acetonitrile), flow rate: 25 mL/min) to give Compound C3-1 (25 mg, 5%) as a solid.

Example 59. Synthesis of Compound 559

[1124] ##STR00333##

[1125] To a solution of C3-1 (320 mg, 0.486 mmol) in MeOH (10 mL) was added Mg powder (349 mg, 14.5 mmol) was added at 60° C. The mixture was stirred at 60° C. for 2 h. Then another portion of Mg powder (349 mg, 14.5 mmol) was added. The final reaction was stirred at 60° C. for 16 hours. The mixture was quenched with HCl (100 mL, 1 M) until the reaction became clear and extracted with DCM (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by a silica gel column (PE/EtOAc=10/1 to 8/1) to give Compound 559 (20 mg, 6%) as a solid.

[1126] .sup.1H NMR (400 MHz, CDCl3) δ 7.52-7.50 (m, 2H), 7.43-7.31 (m, 3H), 5.29-5.28 (m, 1H), 2.43-2.40 (m, 1H), 2.29-2.28 (m, 1H), 2.10-1.60 (m, 7H), 1.52-1.21 (m, 8H), 1.19-0.94 (m, 14H), 0.93-0.91 (m, 5H), 0.63 (d, J=10.8 Hz, 3H).

[1127] LCMS Rt=1.465 min in 2 min chromatography, 10-80 AB, MS ESI calcd. for C.sub.32H.sub.44F.sub.3O [M−H.sub.2O+H].sup.+ 501, found 501.

Example 60. Synthesis of Compound 660, 6051, and 6052

[1128] ##STR00334##

Step 1

[1129] To THF (2 mL) under N.sub.2 at −70° C. was added n-BuLi (1.7 mL, 4.24 mmol). After that, a suspension of CO (500 mg, 1.06 mmol) in THF (5 mL) was added drop-wise to give a suspension. After stirring at −70° C. for 30 min, a solution of D1 (284 mg, 2.12 mmol) in THF (2 mL) was added. Then reaction was stirred at −70° C. for 10 min and then stirred at 20° C. for 16 hours. The reaction mixture was quenched with water (10 mL). The mixture was extracted with EtOAc (3×50 mL). The combined organic phases were washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give crude product which was purified by a silica gel column (PE/EtOAc=10/1) to give D2 (60 mg, 9%) as a solid.

[1130] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.91-7.81 (m, 2H), 7.69-7.49 (m, 3H), 7.33-7.31 (m, 2H), 7.24-7.20 (m, 2H), 7.12-6.98 (m, 1H), 5.32-5.28 (m, 1H), 3.26-3.23 (m, 1H), 2.88-2.65 (m, 2H), 2.57-2.47 (m, 1H), 2.42-2.38 (m, 1H), 2.27-2.07 (m, 1H), 2.04-1.61 (m, 9H), 1.53-1.30 (m, 9H), 1.19-0.93 (m, 11H), 0.93-0.60 (m, 4H), 0.43 (s, 2H).

Step 2

[1131] To a solution of D2 (118 mg, 0.195 mmol) in MeOH (3 mL) was added Mg powder (280 mg, 11.7 mmol) at 60° C. The final reaction was stirred at 60° C. for 16 hours. The mixture was quenched with HCl (100 mL, 1M) until the reaction became clear and extracted with DCM (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by a silica gel column (PE/EtOAc=10/1 to 8/1) to give Compound 660 (32 mg, 35%) as a solid.

[1132] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.32-7.30 (m, 2H), 7.25-7.21 (m, 3H), 5.31-5.29 (m, 1H), 3.77-3.75 (m, 1H), 2.89-2.81 (m, 1H), 2.63-2.57 (m, 1H), 2.44-2.40 (m, 1H), 2.06-1.92 (m, 3H), 1.91-1.60 (m, 5H), 1.58-1.21 (m, 12H), 1.20-0.88 (m, 15H), 0.68 (s, 3H).

[1133] LCMS Rt=1.466 min in 2 min chromatography, 10-80 AB, MS ESI calcd. for C.sub.32H.sub.47O [M−H.sub.2O+H].sup.+ 447, found 447.

Synthesis of 6010, 6051, and 6052

[1134] ##STR00335##

Synthesis of 6010

[1135] ##STR00336##

[1136] To a solution of DA-23-5_1 (400 mg, 1.03 mmol) in THF (6 mL) was added benzylmagnesium bromide (10.3 mL, 10.3 mmol, 1M in THF) at −70° C. under N.sub.2. Then the mixture was stirred at 25° C. for 1 h. The reaction was treated with Sat. NH.sub.4Cl (10 mL), EtOAc (10 mL) and H.sub.2O (5 mL). The mixture was extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine (3×30 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated. The residue was purified by flash column (PE/EA=100/1 to 12/1) to give DA-23-5_2 (6010) (222 mg, 45%) as a solid.

[1137] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.40-7.28 (m, 2H), 7.25-7.15 (m, 3H), 5.30-5.20 (m, 1H), 3.80-3.70 (m, 1H), 2.90-2.30 (m, 3H), 2.05-1.60 (m, 9H), 1.50-1.30 (m, 9H), 1.30-0.90 (m, 16H), 0.90-0.80 (m, 3H), 0.68 (s, 3H).

[1138] LCMS Rt=1.356 min in 2 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. For C.sub.33H.sub.49O [M+H−H.sub.2O].sup.+ 461, found 461.

Synthesis of 6051 and 6052

[1139] ##STR00337##

[1140] DA-23-5_2 (180 mg) was purified by SFC (Column: AD (250 mm*30 mm, 10 um); Condition: 0.1% NH.sub.3H.sub.2O ETOH, 40% B; FlowRate(ml/min): 60) to give impure 6051 (75 mg, 42%) and impure 6052 (80 mg, 45%). The impure 6051(75 mg, 0.156 mmol) was triturated from MeCN (5 mL) at 25° C. to give 6051 (40 mg, 54%) as a solid. The impure 6052 (80 mg) was triturated from MeCN (5 mL) at 25° C. to give 6052 (48 mg, 60%) as a solid. 6051

[1141] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.40-7.28 (m, 2H), 7.25-7.15 (m, 3H), 5.30-5.20 (m, 1H), 3.80-3.70 (m, 1H), 2.90-2.30 (m, 3H), 2.05-1.60 (m, 9H), 1.50-1.30 (m, 9H), 1.30-0.90 (m, 16H), 0.90-0.80 (m, 3H), 0.68 (s, 3H).

[1142] LCMS Rt=1.444 min in 2 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. For C.sub.33H.sub.49O [M+H−H.sub.2O]+ 461, found 461.

[1143] 6052

[1144] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.40-7.28 (m, 2H), 7.25-7.15 (m, 3H), 5.30-5.20 (m, 1H), 3.80-3.70 (m, 1H), 2.90-2.30 (m, 3H), 2.05-1.60 (m, 9H), 1.50-1.30 (m, 9H), 1.30-0.90 (m, 16H), 0.90-0.80 (m, 3H), 0.68 (s, 3H).

[1145] LCMS Rt=1.446 min in 2 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. For C.sub.33H.sub.49O [M+H−H.sub.2O]+ 461, found 461.

Example 61. Syntheses of Compounds 761, 861, and 961

[1146] ##STR00338## ##STR00339##

Step 1

[1147] To a solution of 2,6-di-tert-butyl-4-methylphenol (220 g, 1.0 mol) in toluene (250 mL) was added AlMe.sub.3 (250 mL, 501 mmol, 2 M in toluene) dropwise below 25° C. The solution was stirred at 25° C. for 1 h. Then a solution of E1, synthesis can be found in WO2017007840 (50 g, 167 mmol) in DCM (400 mL) was added dropwise at −78° C. After stirring at −78° C. for 1 h, EtMgBr (167 mL, 501 mmol, 3M in ethyl ether) was added dropwise at −78° C. The resulting solution was stirred at −78° C. to −50° C. for 3 h. The reaction was quenched with saturated citric acid (100 mL) at −78° C. After stirring at 25° C. for 30 min, the resulting mixture was filtered and the filtrate was extracted with DCM (3×100 mL). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude product was combined and purified by a silica gel column (PE/EtOAc=5/1) to give 38 g of the crude product as a solid, which was recrystallized from PE to give E2 as a solid (13.5 g, 13%).

[1148] .sup.1H NMR (CDCl.sub.3) 400 MHz δ 5.33-5.26 (m, 1H), 5.23-5.10 (m, 1H), 2.45-1.90 (m, 6H), 1.78-0.70 (m, 28H).

Step 2

[1149] To a solution of E2 (13 g, 39.5 mmol) and methyl propiolate (8.29 g, 98.7 mmol) in anhydrous DCM (100 mL) under N.sub.2 at 0° C. was added diethylaluminum chloride (1 M in hexane, 158 mL, 158 mmol) dropwise. The mixture was stirred at 20° C. for 16 hours. The reaction mixture was poured into ice-water, extracted with DCM (3×300 mL). The extracts were dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel (PE/EtOAc=5/1) to give E3 (14 g, 86%) as a solid.

[1150] .sup.1H NMR (CDCl.sub.3) 400 MHz δ 6.93 (dd, J=15.6 Hz, 8.0 Hz, 1H), 5.81 (d, J=8.0 Hz, 1H), 5.42-5.38 (m, 1H), 5.33-5.24 (m, 1H), 3.73 (s, 3H), 3.05-2.95 (m, 1H), 2.40-2.30 (m, 1H), 2.10-1.95 (m, 3H), 1.90-1.65 (m, 4H), 1.60-1.25 (m, 9H), 1.88 (d, J=7.2 Hz, 3H), 1.15-0.95 (m, 6H), 0.84 (t, J=7.6 Hz, 3H), 0.78 (s, 3H).

Step 3

[1151] To a solution of E3 (9 g, 21.8 mmol) in THF (100 mL) was added Pd/C (2 g, wet 10%) at 15° C. After degassing and back-fill with H.sub.2 for three times, the reaction mixture was stirred for 16 h at 15° C. with H.sub.2 balloon. The mixture was filtered through a pad of celite. The filtrate was concentrated in vacuo to give crude E4 (8.7 g, crude) as a solid.

[1152] .sup.1H NMR (CDCl.sub.3) 400 MHz δ 5.35-5.25 (m, 1H), 3.69 (s, 3H), 2.40-2.15 (m, 4H), 2.10-1.40 (m, 17H), 2.15-0.80 (m, 16H), 0.70 (s, 3H).

Step 4

[1153] To a solution of E4 (5 g, 12.0 mmol) in THF (100 mL) was added lithium aluminium hydride (1.13 g, 30.0 mmol) at 0° C. Then the reaction was stirred at 25° C. for 5 min. Then the reaction was quenched by aqueous NH.sub.4Cl solution (50 mL) and aqueous citric acid (30 mL) to pH=4-5. Then the reaction solution was extracted with EtOAc (3×100 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give crude product E5 (4 g, 80%) as a solid.

[1154] .sup.1H NMR (CDCl.sub.3) 400 MHz δ 5.35-5.25 (m, 1H), 3.18-3.05 (m, 2H), 2.40-2.32 (m, 1H), 2.08-1.80 (m, 18H), 1.80-0.80 (m, 19H), 0.68 (s, 3H).

Step 5

[1155] To a solution of E5 (1 g, 2.57 mmol) in DCM (15 mL) and THF (15 mL) was added PCC (1.10 g, 5.14 mmol). The resulting reaction mixture was stirred at 25° C. for 2 hours. The combined organic phase was dried, concentrated and purified by flash chromatography (0-15% of EtOAc in PE) to give E6 (700 mg, 70%) as a solid.

[1156] .sup.1H NMR (CDCl.sub.3) 400 MHz δ 9.77 (s, 1H), 5.30-5.26 (m, 1H), 2.46-2.35 (m, 2H), 2.04-1.57 (m, 12H), 1.50-0.83 (m, 23H), 0.68 (m, 3H).

Step 6

[1157] To a solution of E6 (400 mg, 1.03 mmol) in THF (10 mL) was added phenylmagnesium chloride (3.5 mL, 1.5 mmol, 3M in ether) at −70° C. under N.sub.2. Then the mixture was stirred at 20° C. for 20 min. The reaction was treated with saturated NH.sub.4Cl (4 mL), EtOAc (5 mL), and H.sub.2O (3 mL). The mixture was extracted with EtOAc (3×6 mL). The combined organic layers were washed with brine (2×15 mL), dried over Na.sub.2SO4, filtered, concentrated in vacuum, purified by column chromatography on silica gel (PE/EA=30/1 to 10/1) to give Compound 761 (300 mg, 62%) as a solid.

[1158] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.40-7.30 (m, 4H), 7.30-7.20 (m, 1H), 5.32-5.26 (m, 1H), 4.64-4.52 (m, 1H), 2.38-2.34 (m, 1H), 2.06-1.88 (m, 3H), 1.86-1.15 (m, 24H), 1.12-1.00 (m, 3H), 1.00-0.78 (m, 7H), 0.66 (s, 3H).

[1159] LCMS Rt=1.303 min in 2 min chromatography, 30-90 AB, MS ESI calcd. For C.sub.32H.sub.45 [M+H−2H.sub.2O].sup.+429, found 429.

Step 7

[1160] Compound 761 (300 mg) was purified by SFC (Column: OD (250 mm*30 mm, 10 um); Condition: 0.1% NH.sub.3—H.sub.2O ETOH, 40% B; Flow Rate (ml/min): 60) to give Compound 861 (40 mg, 22%) as a solid and Compound 10 (59 mg, 33%) as a solid.

[1161] Compound 861:

[1162] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.39-7.30 (m, 4H), 7.30-7.20 (m, 1H), 5.32-5.26 (m, 1H), 4.64-4.52 (m, 1H), 2.39-2.34 (m, 1H), 2.06-1.90 (m, 3H), 1.86-1.15 (m, 24H), 1.12-1.00 (m, 3H), 1.00-0.80 (m, 7H), 0.66 (s, 3H).

[1163] LCMS Rt=1.413 min in 2 min chromatography, 30-90AB 2MIN_E, MS ESI calcd. For C.sub.32H.sub.45 [M+H−2H.sub.2O].sup.+429, found 429.

[1164] Compound 961:

[1165] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.39-7.30 (m, 4H), 7.30-7.20 (m, 1H), 5.31-5.26 (m, 1H), 4.64-4.52 (m, 1H), 2.39-2.34 (m, 1H), 2.06-1.90 (m, 3H), 1.86-1.35 (m, 17H), 1.28-1.20 (m, 2H), 1.20-0.80 (m, 13H), 0.66 (s, 3H).

[1166] LCMS Rt=1.425 min in 2 min chromatography, 30-90AB, MS ESI calcd. For C.sub.32H.sub.45 [M+H−2H.sub.2O].sup.+429, found 429.

Synthesis of 861 to Determine Stereochemistry

[1167] ##STR00340##

Synthesis of ST-200-095-001_1

[1168] ##STR00341##

[1169] To a freshly distilled anhydrous THF (1 mL) was added n-BuLi (2.47 mL 6.18 mmol, 2.5 M in n-hexane) drop-wise at −70° C. Then a solution of ST-200-550_15 (1 g, 2.06 mmol) in anhydrous THF (10 mL) was drop-wise. After stirring at −70° C. for 1 h, (2S)-2-phenyloxirane (371 mg, 3.09 mmol) was added at −70° C. and the reaction was stirred for another 1 h. The reaction mixture was stirred at 25° C. (room temperature) for 12 h. The reaction was quenched by saturated NH.sub.4C1.aq (100 mL). The aqueous phase was extracted with EtOAc (3×50 mL). The combine organic phase was washed with saturated brine (2×50 mL), drive over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give ST-200-095-001_1 (1.1 g, crude) as an oil, which was used directly for the next step.

Synthesis of ST-200-095-001

[1170] ##STR00342##

[1171] To a solution of ST-200-095-001_1 (1.1 g, crude) in MeOH (100 mL) was added Mg powder (2.17 g, 90.5 mmol) and NiCl.sub.2 (40 mg) at 25° C. under N.sub.2. After stirring at 60° C. for 1 h under N.sub.2, the reaction mixture was quenched with HCl (300 mL, 1 M) until the reaction became clear. The aqueous phase was extracted with EtOAc (3×100 mL). The combined organic phase was washed with saturated brine (2×100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=8/1 to 5/1) to afford ST-200-095-001 (0.65 g, 77%, impure) as a solid.

[1172] The ST-200-095-001(650 mg, 1.39 mmol) was re-crystallized from MeCN (40 mL) to give ST-200-095-001 (580 mg, 69%, impure) as a solid.

[1173] The ST-200-095-001 (300 mg, 0.6455 mmol) was purified by SFC (Instrument: SFC-16, Column: OD (250 mm*30 mm, 5 um), Condition: 0.1% NH.sub.3H.sub.2O ETOH, Begin B: 45%, End B: 45%, FlowRate(ml/min): 60, Injections: 70) to give ST-200-095-001 (256 mg, 59%) as a solid.

[1174] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.88-7.81 (m, 4H), 7.80-7.23 (m, 3H), 5.32-5.25 (m, 1H), 4.65-4.61 (m, 1H), 2.40-2.30 (m, 1H), 2.06-1.90 (m, 3H), 1.88-1.64 (m, 5H), 1.51-1.05 (m, 15H), 1.04-0.79 (m, 12H), 0.66 (s, 3H).

[1175] LCMS Rt=1.287 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.32H.sub.45 [M+H−2H.sub.2O].sup.+ 429, found 429.

Example 63: Synthesis of 6347 and 6348

[1176] ##STR00343##

Synthesis of DA-23-3_1 (aka DA-28-1)

[1177] ##STR00344##

[1178] To a solution of 001-4 (50 g, 128 mmol) in DCM (800 mL) was added DMP (108 g, 256 mmol) at 30° C. The reaction mixture was stirred at 30° C. for 10 minutes. And H.sub.2O (2.3 g, 128 mmol) was added dropwise. The reaction mixture was quenched with Saturated NaHCO.sub.3 aqueous (500 mL) until pH of the aqueous layer became about 9. The mixture was filtered. The DCM layer was separated and the aqueous phase was extracted with DCM (100 mL). The combined organic phase was washed with saturated Na.sub.2S.sub.2O.sub.3 aqueous (600 mL), brine (500 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to give DA-28-1_1 (108 g, crude) as an oil. The reaction was conducted in parallel for 2 times.

[1179] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.30-5.26 (m, 1H), 3.67 (s, 3H), 3.30-3.22 (m, 1H), 2.85-2.79 (m, 1H), 2.50-2.15 (m, 4H), 2.08-1.96 (m, 3H), 1.90-1.71 (m, 2H), 1.56-1.45 (m, 6H), 144-1.19 (m, 3H), 1.17 (s, 3H), 1.15-0.97 (m, 5H), 0.96-0.88 (m, 3H), 0.70 (s, 3H).

[1180] To a solution of BHT (367 g, 1.67 mmol) in toluene (1000 mL) under nitrogen at 0° C. was added trimethylaluminum (2 M in toluene, 418 mL, 837 mmol) dropwise. The mixture was stirred at 0° C. for 30 min and used directly as a solution of MAD (0.59 M in toluene) without further purification. To the solution of MAD (0.59 M in toluene, 1410 mL, 837 mmol) under nitrogen at −78° C. was added a solution of DA-28-1_1 (108 g, 279 mmol) in toluene (500 mL) dropwise. The mixture was stirred at −78° C. for 30 min. EtMgBr (3 M in diethyl ether, 278 mL, 837 mmol, 3M in ether) was added dropwise. The resulting mixture was stirred at −78° C. for 1 hr. The reaction mixture was poured to ice-cooled aqueous citric acid (1000 mL), extracted with EtOAc (2×500 mL). The combined organic layer was washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel (0-20% of EtOAc in PE) to give DA-28-1_2 (95 g, impure) as an oil. The reaction was conducted in parallel for 2 times.

[1181] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.30-5.26 (m, 1H), 3.65 (s, 3H), 2.48-2.18 (m, 4H), 2.08-1.91 (m, 2H), 1.90-1.76 (m, 4H), 1.75-1.61 (m, 4H), 1.60-1.48 (m, 5H), 1.47-1.22 (m, 5H), 1.17 (s, 1H), 1.16-1.02 (m, 3H), 1.01-0.96 (m, 2H), 0.95-0.90 (m, 1H), 0.89-0.82 (m, 4H), 0.81-0.76 (m, 2H), 0.67 (s, 3H).

[1182] To a solution of DA-28-1_2 (60 g, 144 mmol) in THF (1200 mL) under N.sub.2 at 0° C. was added LiAlH.sub.4 (8.19 g, 216 mmol) in portions. The reaction was stirred at 20° C. for 30 min. The reaction was quenched with 2 M HCl (600 mL) at 0° C. till pH of the aqueous phase was about 2. The mixture was filtered. The solid was collected and the aqueous layer was extracted with EtOAc (2×400 mL). The combined organic phase was washed with sat. NaHCO.sub.3 (600 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude product, which was purified with the solid by trituration with MeCN (800 mL) to give DA-28-1_3 (45 g, 81%) as a solid.

[1183] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.30-5.26 (m, 1H), 3.65-3.56 (m, 2H), 2.40-2.32 (m, 1H), 2.10-2.00 (m, 1H), 1.99-1.90 (m, 2H), 1.89-1.58 (m, 5H), 1.56-1.31 (m, 10H), 1.30-1.19 (m, 3H), 1.18-1.03 (m, 8H), 1.02-0.88 (m, 5H), 0.87-0.78 (m, 3H), 0.68 (s, 3H). To a solution of DA-28-1_3 (45 g, 115 mmol) in DCM (800 mL) was added DMP (97.5 g, 230 mmol) at 20° C. The reaction mixture was stirred at 20° C. for 10 min. The reaction mixture was quenched with Saturated NaHCO.sub.3 aqueous (500 mL) at 20° C. The mixture was filtered. The DCM layer was separated and the aqueous phase was extracted with DCM (200 mL). The combined organic phase was washed with saturated Na.sub.2S.sub.2O.sub.3 aqueous (3×500 mL), sat. NaHCO.sub.3 (500 mL), water (500 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated. Combined with another batch from 25 g of DA-28-13, the crude product was triturated with MeCN (500 mL) to give DA-028-1 (48 g) as a solid. 500 mg of impure DA-28-1 was purified by flash column (0-15% of EtOAc in PE) to give DA-28-1 (194 mg, 39%) as a solid.

[1184] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.77 (s, 1H), 5.30-5.27 (m, 1H), 2.50-2.32 (m, 3H), 2.05-1.94 (m, 3H), 1.93-1.67 (m, 3H), 1.66-1.58 (m, 3H), 1.56-1.22 (m, 10H), 1.20-1.04 (m, 4H), 1.02 (s, 3H), 1.00-0.90 (m, 5H), 0.89-0.80 (m, 3H), 0.68 (s, 3H).

[1185] LCMS Rt=1.229 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.26H.sub.41O [M+H−H.sub.2O].sup.+ 369, found 369.

Synthesis of DA-23-7_1

[1186] ##STR00345##

[1187] Pyridin-2-yllithium (437 mg, 5.15 mmol) was added to a solution of DA-23-3_1 (400 mg, 1.03 mmol) in THF (3 mL) at −70° C. The mixture was stirred at 25° C. for 1 hr. The mixture was quenched with Sat. NH.sub.4Cl (20 mL), extracted with EtOAc (3×15 mL). The combined organic layers were washed with brine (30 mL×3), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum to give a crude product, which was purified by silica gel chromatography (PE/EtOAc=30/1 to 6/1) to afford DA-23-7_1 (200 mg, 42%) as a solid.

[1188] .sup.1H NMR CDCl.sub.3 Bruker_P_400 MHz δ 8.59-8.55 (m, 1H), 7.72-7.64 (m, 1H), 7.25-7.16 (m, 2H), 5.32-5.24 (m, 1H), 4.75-4.62 (m, 1H), 4.10-4.00 (m, 1H), 2.40-2.30 (m, 1H), 2.07-1.30 (m, 16H), 1.30-0.80 (m, 20H), 0.65 (s, 3H).

Synthesis of 6347 and 6348

[1189] ##STR00346##

(Stereochemistry Randomly Assigned).

[1190] The compound DA-23-7_1 (510 mg, 1.08 mmol) was purified by SFC (Column: Chiralpak AS-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO.sub.2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.) to afford 6348 (60 mg, 30%) as solid and 6347 (60 mg, 30%) as a solid.

[1191] 6348

[1192] .sup.1H NMR CDCl.sub.3 Bruker_P_400 MHz δ 8.59-8.51 (m, 1H), 7.72-7.64 (m, 1H), 7.25-7.17 (m, 2H), 5.32-5.24 (m, 1H), 4.73-4.62 (m, 1H), 4.05-4.01 (m, 1H), 2.40-2.31 (m, 1H), 2.07-1.67 (m, 6H), 1.66-1.57 (m, 3H), 1.50-1.29 (m, 9H), 1.28-1.19 (m, 2H), 1.18-1.03 (m, 5H), 1.03-1.00 (m, 3H), 1.00-0.94 (m, 1H), 0.94-0.88 (m, 4H), 0.88-0.82 (m, 3H), 0.68 (s, 3H).

[1193] LCMS Rt=0.839 min in 2 min chromatography, 30-90AB, purity 99%, MS ESI calcd. For C.sub.21H.sub.48NO.sub.2 [M+H].sup.+ 466, found 466.

[1194] 6347

[1195] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.57-8.51 (m, 1H), 7.71-7.65 (m, 1H), 7.25-7.17 (m, 2H), 5.35-5.23 (m, 1H), 4.75-4.67 (m, 1H), 4.12-4.06 (m, 1H), 2.41-2.31 (m, 1H), 2.08-1.91 (m, 3H), 1.88-1.67 (m, 4H), 1.67-1.58 (m, 1H), 1.50-1.32 (m, 6H), 1.31-1.14 (m, 4H), 1.13-1.03 (m, 4H), 1.03-1.01 (m, 2H), 1.01-0.93 (m, 2H), 0.93-0.89 (m, 4H), 0.89-0.81 (m, 4H) 0.68 (s, 3H).

[1196] LCMS Rt=0.825 min in 2 min chromatography, 30-90AB, purity 99%, MS ESI calcd. For C.sub.21H.sub.48NO.sub.2 [M+H].sup.+ 466, found 466.

Example 64: Synthesis of 6457, 6458, and 6459

[1197] ##STR00347##

[1198] Stereochemistry assigned based on NMR data.

[1199] The experimental of intermediate DA-23-3_1 could be found in Example 63.

Synthesis of DA-23-3_2 (6457)

[1200] ##STR00348##

[1201] Isopropylmagnesium chloride (20.6 mL, 41.2 mmol, 2 M in THF) was added to a suspension of 3-bromopyridine (6.50 g, 41.2 mmol) in THF (10 mL) at 0° C. The mixture was stirred at 25° C. for 1 h. After cooling to 0° C., a solution of DA-23-3_1 (800 mg, 2.06 mmol) in THF (10 mL) was added. The mixture was stirred at 25° C. for 2 h. To the mixture was added NH.sub.4Cl (50 mL, 10% aq.). Combined with another batch from 100 mg of DA-23-31, the mixture was extracted with EtOAc (2×50 mL). The organic layer was separated. The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated. The residue was purified by flash column (0-100% of EtOAc in PE) to afford impure DA-23-3_2 (560 mg, 58%) as a solid. The impure DA-23-3_2 (560 mg, 1.20 mmol) was triturated from MeCN (20 mL) at 25° C. to give DA-23-3_2 (406 mg, 73%) as a solid.

[1202] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.60-8.50 (m, 2H), 7.72-7.68 (m, 1H), 7.28-7.17 (m, 1H), 5.32-5.25 (m, 1H), 4.22-4.10 (m, 1H), 2.40-2.30 (m, 1H), 2.10-1.92 (m, 4H), 1.92-1.68 (m, 4H), 1.68-1.53 (m, 6H), 1.53-1.35 (m, 4H), 1.35-1.01 (m, 10H), 1.01-0.80 (m, 9H), 0.66 (s, 3H).

[1203] LCMS Rt=0.921 min in 2 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. For C.sub.31H.sub.48NO.sub.2 [M+H].sup.+ 466, found 466.

Synthesis of 6458 and 6459

[1204] ##STR00349##

[1205] DA-23-3_2 (378 mg, 0.811 mmol) was purified by SFC (column: AD (250 mm*30 mm, 10 um), gradient: 40-40% B (A=0.1% NH.sub.3H.sub.2O IPA), flow rate: 60 mL/min) to give DA-23-9 (110 mg, 29%) as a solid and DA-23-10 (120 mg, 31.8%) as a solid.

[1206] 6458

[1207] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.55-8.45 (m, 2H), 7.70-7.60 (m, 1H), 7.25-7.18 (m, 1H), 5.25-5.18 (m, 1H), 4.15-4.05 (m, 1H), 2.30-2.20 (m, 1H), 2.10-1.85 (m, 4H), 1.90-1.62 (m, 5H), 1.52-1.28 (m, 10H), 1.28-0.95 (m, 9H), 0.95-0.80 (m, 6H), 0.80-0.75 (m, 3H), 0.59 (s, 3H).

[1208] LCMS Rt=0.926 min in 2 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. For C.sub.31H.sub.48NO.sub.2 [M+H].sup.+ 466, found 466.

[1209] SFC_E1 Rt=1.603 min in 10 min chromatography, AD_3_IPA_DEA_40_25ML, 100% de. 6459

[1210] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.55-8.45 (m, 2H), 7.70-7.60 (m, 1H), 7.25-7.18 (m, 1H), 5.25-5.18 (m, 1H), 4.15-4.05 (m, 1H), 2.30-2.20 (m, 1H), 2.01-1.65 (m, 7H), 1.61-1.53 (m, 3H), 1.53-1.29 (m, 8H), 1.25-1.12 (m, 4H), 1.12-0.88 (m, 12H), 0.88-0.75 (m, 3H), 0.59 (s, 3H).

[1211] LCMS Rt=0.928 min in 2 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. For C.sub.31H.sub.48NO.sub.2 [M+H].sup.+ 466, found 466.

[1212] SFC_E1 Rt=2.037 min in 10 min chromatography, AD_3_IPA_DEA_40_25ML, 98% de.

Example 65: Synthesis of 6544, 6571, and 6572

[1213] ##STR00350##

[1214] The experimental of intermediate DA-23-3_4 can be found in Example 67.

Synthesis of DA-23-3_5

[1215] ##STR00351##

[1216] PhLi (1.71 mL, 1.5 M in ether, 2.57 mmol) was added to a solution of DA-23-3_4 (200 mg, 0.514 mmol) in THF (5 mL). The mixture was stirred at 25° C. for 4 h. After cooling, the mixture was treated with NH.sub.4Cl (10 mL, sat.). The mixture was extracted with EtOAc (20 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give an oil. The mixture was purified by silica gel chromatography (PE/EtOAc=0 to 4/1) to afford DA-23-3_5 (110 mg, 46.0%) as a solid. DA-23-3_5 (20 mg) was used to delivery.

[1217] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 5H), 4.61 (brs, 1H), 1.95-1.88 (m, 1H), 1.80-1.50 (m, 8H), 1.50-1.15 (m, 13H), 1.15-0.83 (m, 15H), 0.82 (s, 3H), 0.68-0.55 (m, 4H).

[1218] LCMS Rt=1.286 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.47[M+H−2H.sub.2O].sup.+ 431, found 431.

Synthesis of 6571 and 6572

[1219] ##STR00352##

[1220] DA-23-3_5 (90 mg, 192 umol) was purified by SFC (Column: AD (150×4.6 mm, 3 um), Gradient: 5%-40% B (A: CO.sub.2 B: ethanol) Flow rate: 2.5 mL/min) to afford DA-23-13 (6 mg, 7%) as a solid and DA-23-14 (8 mg, 9%) as a solid.

[1221] 6571

[1222] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 5H), 4.62 (brs, 1H), 2.00-1.75 (m, 8H), 1.50-1.30 (m, 8H), 1.30-1.05 (m, 10H), 1.05-0.83 (m, 10H), 0.82 (s, 3H), 0.68-0.55 (m, 4H).

[1223] LCMS Rt=1.262 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.47[M+H−2H.sub.2O].sup.+ 431, found 431.

[1224] 6572

[1225] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 5H), 4.62 (brs, 1H), 1.95-1.88 (m, 1H), 1.80-1.50 (m, 9H), 1.50-1.15 (m, 13H), 1.15-0.83 (m, 14H), 0.81 (s, 3H), 0.68-0.55 (m, 4H).

[1226] LCMS Rt=1.258 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.47 [M+H−2H.sub.2O].sup.+ 431, found 431.

Synthesis of 6571 to Confirm Stereochemistry at C24

[1227] ##STR00353##

[1228] To a solution of DA-27-5 (400 mg, 0.8217 mmol) in anhydrous THF (3 mL) was added n-BuLi (0.984 mL, 2.46 mmol, 2.5M in n-hexane) drop-wise at −70° C. under N.sub.2. After stirring at −70° C. for 30 min. a solution of (2S)-2-phenyloxirane (147 mg, 1.23 mmol) in anhydrous THF (0.5 mL) was added drop-wise at −70° C. The reaction mixture was stirred at −70° C. for another 1 h and then stirred at 25° C. (room temperature) for 12 h. The reaction was quenched by saturated NH.sub.4C1.aq (50 mL). The aqueous phase was extracted with EtOAc (3×50 mL). The combine organic phase was washed with saturated brine (2×50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give ST-200-095-008_1 (0.5 g, crude) as an oil. The crude residue was directly used the next step.

[1229] To a solution of ST-200-095-008_1 (0.5 g, crude) in MeOH (50 mL) was added Mg powder (0.986 g, 41.1 mmol) and NiCl.sub.2 (20 mg) at 25° C. under N.sub.2. After stirring at 60° C. for 1 h, the reaction mixture was quenched with HCl (100 mL, 1 M) until the reaction became clear. The aqueous phase was extracted with EtOAc (2×100 mL). The combined organic phase was washed with saturated NaHCO.sub.3.aq (2×50 mL) and washed whit brine (2×100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=8/1 to 5/1) to afford ST-200-095-008 (250 mg, impure) as a solid, which was re-crystallized from MeCN (20 mL) at 82° C. reflux for 30 mins. The mixture stirred was cool to 25° C. (room temperature). The suspension was filtration in vacuum to get ST-200-095-008 (200 mg, impure) as a solid. The ST-200-095-008 (200 mg, 0.428 mmol) was by SFC (Instrument: SFC-16, Column: OD (250 mm*30 mm, 5 um), Condition: 0.1% NH.sub.3H.sub.2O ETOH, Begin B: 45%, End B: 45%, FlowRate(ml/min): 60, Injections: 70) to give ST-200-095-008 (148 mg, 38%) as a solid.

[1230] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.38-7.31 (m, 4H), 7.30-7.24 (m, 2H), 4.58-4.54 (m, 1H), 1.96-1.73 (m, 4H), 1.67-1.59 (m, 4H), 1.54-1.14 (m, 15H), 1.13-0.86 (m, 13H), 0.84-0.80 (m, 3H), 0.68-0.59 (m, 4H).

[1231] LCMS Rt=1.306 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.32H.sub.47 [M+H−2H.sub.2O].sup.+ 431, found 431.

[1232] SFC Rt=5.523 min in 10 min chromatography, OD_3_EtOH_DEA_5_40_25ML (“Column: Chiralcel OD-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.”), 100% de.

Example 66: Synthesis of 6680 and 6681

[1233] ##STR00354##

[1234] The stereochemistry at C24 is randomly assigned.

[1235] The experimental of intermediate E-2761 can be found in Example 63 or Example 60.

Synthesis of DA-62-4_1

[1236] ##STR00355##

[1237] Isopropylmagnesium chloride (2 M, 2.59 mL) was added dropwise to a solution of l-bromo-4-cyanobenzene (940 mg, 5.19 mmol) in THF (10 mL) at 0° C. under N.sub.2. After stirring at 0° C. for 2 h, a solution of E-2761 (200 mg, 0.517 mmol) in THF (10 mL) was added at 0° C. under N.sub.2. The mixture was stirred at 0° C. for 2 h and treated with saturated aqueous NH.sub.4Cl (30 mL). The aqueous phase was extracted with EtOAc (3×20 mL). The combined organic phase was washed with saturated brine (2×200 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give an oil, which was purified by flash column (0˜20% of EtOAc in PE) to give DA-62-4_1 (140 mg, 55%) as a solid.

Synthesis of 6680 and 6681

[1238] ##STR00356##

[1239] DA-62-4_1 (140 mg, 0.285 mmol) was purified by SFC (Column: AD (150×4.6 mm, 3 um), Gradient: 5%-40% B (A: CO.sub.2 B: ethanol) Flow rate: 2.5 mL/min) to afford DA-62-4(32.0 mg, 23%) as a solid and DA-62-10 (33.0 mg, 24%) as a solid. The chiral center at C24 was assigned randomly.

[1240] 6680:

[1241] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.68-7.60 (m, 2H), 7.50-7.40 (m, 2H), 5.30-5.25 (m, 1H), 4.72-4.62 (m, 1H), 2.40-2.30 (m, 1H), 2.08-1.85 (m, 4H), 1.80-1.55 (m, 6H), 1.50-1.32 (m, 8H), 1.25-1.00 (m, 10H), 1.00-0.75 (m, 9H), 0.66 (s, 3H).

[1242] LCMS Rt=1.170 min in 2.0 min chromatography, 30-90 AB, purity 98%, MS ESI calcd. for C.sub.33H.sub.46NO [M+H−H.sub.2O].sup.+ 472, found 472.

[1243] 6681

[1244] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.68-7.60 (m, 2H), 7.50-7.40 (m, 2H), 5.30-5.25 (m, 1H), 4.72-4.62 (m, 1H), 2.40-2.30 (m, 1H), 2.08-1.85 (m, 4H), 1.80-1.55 (m, 6H), 1.50-1.32 (m, 8H), 1.25-1.00 (m, 10H), 1.00-0.75 (m, 9H), 0.66 (s, 3H).

[1245] LCMS Rt=1.174 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.33H.sub.46NO [M+H−H.sub.2O].sup.+ 472, found 472.

Example 67: Synthesis of 6754 and 6755

[1246] ##STR00357## ##STR00358##

[1247] The stereochemistry for 6754 and 6755 has been assigned based on NMR data.

[1248] The synthesis of ST-200-3ET-B12_1 can be found in Example 125.

Synthesis of DA-23-3_3

[1249] ##STR00359##

[1250] LiAlH.sub.4 (198 mg, 2.54 mmol) was added in three portions to a solution of ST-200-3ET-B12_1 (1.1 g, 2.62 mmol) in THF (10 mL) at 0° C. under N.sub.2. After stirring at 20° C. for 1 hour, the mixture was quenched with water (10 mL) at 0° C., followed by adding HCl (10 mL, 1 mol/L). The aqueous phase was extracted with EtOAc (2×10 mL). The combined organic phase was washed with saturated brine (2×10 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-50% of EtOAc in PE) to give DA-23-3_3 (1 g, 98%) as a solid.

[1251] .sup.1H NMR CDCl.sub.3 400 MHz δ 3.65-3.55 (m, 2H), 1.98-1.92 (m, 1H), 1.88-1.75 (m, 1H), 1.70-1.40 (m, 13H), 1.40-1.19 (m, 7H), 1.19-0.98 (m, 7H), 0.98-0.80 (m, 11H), 0.66-0.61 (m, 4H).

Synthesis of DA-23-3_4

[1252] ##STR00360##

[1253] To a solution of DA-23-3_3 (1 g, 2.55 mmol) in anhydrous DCM (30 mL) was added silica gel (1 g) and PCC (1.09 g, 5.10 mmol). After stirring at 20° C. for 1 hours, the reaction mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography on silica gel (PE/EA=50/1 to 10/1) to give DA-23-3_4 (600 mg, 60%) as a solid. NMR CDCl.sub.3 400 MHz δ 9.98-9.97 (m, 1H), 2.50-2.20 (m, 2H), 2.05-1.50 (m, 3H), 1.50-1.19 (m, 15H), 1.19-0.99 (m, 7H), 0.99-0.82 (m, 12H), 0.70-0.55 (m, 4H).

Synthesis of DA-23-3_5

[1254] ##STR00361##

[1255] Isopropylmagnesium chloride (7.70 mL, 15.4 mmol, 2 M in THF) was added to a suspension of 3-bromopyridine (2.43 g, 15.4 mmol) in THF (10 mL) at 0° C. The mixture was stirred at 25° C. for 1 hour. To the fresh prepared pyridin-3-ylmagnesium chloride (2.12 g, 15.4 mmol) was added DA-23-3_4 (300 mg, 0.771 mmol) in THF (10 mL) at 0° C. The mixture was stirred at 25° C. for 2 hours and quenched with NH.sub.4Cl (20 mL, 10% aq.). The mixture was extracted with EtOAc (2×20 mL). The organic layer was separated. The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated to afford DA-23-3_5 (280 mg, crude) as a solid.

Synthesis of 6754 and 6755

[1256] ##STR00362##

[1257] 190 mg of DA-23-3_5 was separated with SFC (Column: AD (250 mm*30 mm, 10 um); Condition: 0.1% NH.sub.3H.sub.2O IPA, 40% B; FlowRate(ml/min): 60;) to give 6754 (34 mg, impure) and 6755 (35 mg, impure) as a solid.

[1258] 34 mg of impure 6754 was re-crystallized from MeCN (5 mL) at 70° C. to give 6754 (14 mg) as a solid.

[1259] 35 mg of impure 6755 was re-crystallized from MeCN (5 mL) at 70° C. to give 6755 (19 mg) as a solid.

[1260] 6457:

[1261] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.65-8.50 (m, 2H), 7.75-7.60 (m, 1H), 7.35-7.27 (m, 1H), 4.75-4.60 (m, 1H), 2.00-1.60 (m, 8H), 1.55-1.15 (m, 14H), 1.10-0.75 (m, 18H), 0.70-0.50 (m, 4H).

[1262] LCMS Rt=0.846 min in 2 min chromatography, 30-90AB_E, purity 99.8%, MS ESI calcd. For C.sub.31H.sub.50NO.sub.2 [M+H].sup.+ 468, found 468.

[1263] 6755

[1264] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.65-8.50 (m, 2H), 7.75-7.60 (m, 1H), 7.35-7.27 (m, 1H), 4.75-4.60 (m, 1H), 2.00-1.60 (m, 8H), 1.55-1.15 (m, 15H), 1.10-0.75 (m, 17H), 0.70-0.50 (m, 4H).

[1265] LCMS Rt=0.836 min in 2 min chromatography, 30-90AB_E, purity 98.3%, MS ESI calcd. For C.sub.31H.sub.50NO.sub.2 [M+H].sup.+ 468, found 468.

Example 68: Synthesis of 6895 and 6896

[1266] ##STR00363##

[1267] The experimental of intermediate E-2761 can be found in Example 63.

Synthesis of DA-62-2_1

[1268] ##STR00364##

[1269] 1-bromo-3-fluorobenzene (900 mg, 5.14 mmol) was added to a suspension of magnesium (124 mg, 5.14 mmol) and a small amount of iodine (130 mg, 0.514 mmol) in tetrahydrofuran (3 mL). After stirring for 2 h at 50° C., a solution E-2761 (200 mg, 0.517 mmol) in THF (10 mL) was added at 15° C. under N.sub.2. The mixture was stirred at 15° C. for 2 h and quenched with saturated aqueous NH.sub.4Cl (30 mL). The aqueous phase was extracted with EtOAc (3×20 mL). The combined organic phase was washed with saturated brine (2×20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give an oil. The mixture was purified by flash column (0-20% of EtOAc in PE) to give DA-62-2_1 (120 mg, 48%) as a solid.

[1270] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 1H), 7.10-7.02 (m, 2H), 7.00-6.90 (m, 1H), 5.30-5.25 (m, 1H), 4.68-4.55 (m, 1H), 2.40-2.30 (m, 1H), 2.05-1.90 (m, 3H), 1.90-1.50 (m, 7H), 1.50-1.30 (m, 8H), 1.30-1.15 (m, 3H), 1.15-0.86 (m, 13H), 0.86-0.80 (m, 3H), 0.66 (s, 3H).

Synthesis of 6895 and 6896

[1271] ##STR00365##

[1272] DA-62-2_1 (120 mg, 0.248 mmol) was purified by SFC (Column: AD (150×4.6 mm, 3 um), Gradient: 5%-40% B (A: CO.sub.2 B: ethanol) Flow rate: 2.5 mL/min) to afford 6895(32.0 mg, 27%) as a solid and 6896 (40.0 mg, 34%) as a solid.

[1273] 6895:

[1274] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 1H), 7.10-7.02 (m, 2H), 7.00-6.90 (m, 1H), 5.30-5.25 (m, 1H), 4.68-4.55 (m, 1H), 2.40-2.30 (m, 1H), 2.05-1.90 (m, 3H), 1.90-1.50 (m, 7H), 1.50-1.30 (m, 8H), 1.30-1.13 (m, 4H), 1.13-0.86 (m, 12H), 0.86-0.78 (m, 3H), 0.66 (s, 3H).

[1275] LCMS Rt=1.263 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.44F [M+H−2H.sub.2O].sup.+ 447, found 447.

[1276] 6896:

[1277] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 1H), 7.10-7.02 (m, 2H), 7.00-6.90 (m, 1H), 5.30-5.25 (m, 1H), 4.65-4.53 (m, 1H), 2.40-2.30 (m, 1H), 2.05-1.90 (m, 3H), 1.90-1.50 (m, 7H), 1.50-1.30 (m, 8H), 1.30-0.86 (m, 16H), 0.86-0.80 (m, 3H), 0.66 (s, 3H).

[1278] LCMS Rt=1.261 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.44F [M+H−2H.sub.2O].sup.+ 447, found 447.

Synthesis of 6896 to Confirm Stereochemistry

[1279] ##STR00366##

[1280] To a mixture of trimethylsulfoxonium iodide (106 g, 482 mmol) in DMSO (200 mL) and THF (150 mL) was added t-BuOK (53.9 g, 482 mmol). The mixture was stirred at 40° C. for 1 hour. The solution was then cooled to 0° C. and ST-200-095-005_1M (30 g, 241 mmol) in THF (50 mL) was added at 0° C. The reaction mixture was stirred 30 minutes and poured into H.sub.2O (300 mL). The resulting mixture was extracted with EtOAc (3×200 mL). The combined organic layers were washed with H.sub.2O (2×100 mL), brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0˜1% of EtOAc in PE) to give product ST-200-095-005_2M (32 g, 96%) as an oil.

[1281] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 1H), 7.10-7.01 (m, 1H), 7.01-6.80 (m, 2H), 3.80-3.78 (m, 1H), 3.20-3.10 (m, 1H), 2.75-2.70 (m, 1H).

[1282] To a solution of R,R-cat (86.9 mg, 0.144 mmol) in toluene (5 mL) was added AcOH (88.8 mg, 1.48 mmol). The mixture was stirred at 25° C. open to air for 30 min and concentrated in vacuum to leave a crude solid. The resulting catalyst residue was dissolved in ST-200-095-005_2M (5 g, 36.1 mmol) at 25° C. The reaction flask was cooled to 0° C., and H.sub.2O (356 mg, 19.8 mmol) was added dropwise over 5 min. The reaction was allowed to warm to 25° C. and stirred 16 hrs. The reaction mixture was purified directly by silica gel chromatography (PE %=100%) to afford ST-200-095-005_3M (2 g, 40%) as an oil. The ee % was 100%.

[1283] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 1H), 7.10-7.01 (m, 1H), 7.01-6.80 (m, 2H), 3.80-3.78 (m, 1H), 3.20-3.10 (m, 1H), 2.75-2.70 (m, 1H).

[1284] To a THF (1 mL) under N.sub.2 at −70° C. was added n-BuLi (2.5 M, 3.09 mmol, 1.23 mL). After that, a solution of ST-200-095-005_3 (300 mg, 0.618 mmol) in THF (2 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, a solution of ST-200-095-005_3M (341 mg, 2.47 mmol) in THF (2 mL) was added. Then reaction was stirred at stirred at 25° C. for 16 hours. The mixture was poured into ice-water (20 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to afford ST-200-095-005_4M (500 mg, crude) as a solid, which was used directly for the next step.

[1285] To a solution of ST-200-095-005_4M (500 mg, 0.802 mmol) in MeOH (100 mL) was added NiCl.sub.2 (5 mg) and Mg powder (768 mg, 32.0 mmol) at 65° C. in four portions. The reaction mixture was cooled to 25° C. and quenched by saturated aqueous NH.sub.4Cl (100 mL). The mixture was stirred for 1 hour. The resulting mixture was extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-10% of EtOAc in PE) and re-crystallized from DCM/n-hexane (0.5 mL/10 mL) at 25° C. to give ST-200-095-005_5M (20 mg, 29%) as a solid.

[1286] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.27 (m, 1H), 7.15-7.01 (m, 1H), 7.01-6.80 (m, 2H), 5.27-5.25 (m, 1H), 4.65-4.60 (m, 1H), 2.40-2.30 (m, 1H), 2.02-1.85 (m, 3H), 1.75-1.65 (m, 7H), 1.65-1.25 (m, 7H), 1.25-1.01 (m, 10H), 0.66 (s, 3H).

[1287] LCMS Rt=1.287 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.32H.sub.44FO [M+H−2H.sub.2O].sup.+ 447, found 447.

Example 69: Synthesis of 6997 and 6998

[1288] ##STR00367##

[1289] Stereochemistry at C24 was assigned based on NMR data.

[1290] The experimental of intermediate E-2761 can be found in Example 63.

Synthesis of DA-62-5_1

[1291] ##STR00368##

[1292] Isopropylmagnesium chloride (2 M, 2.58 mL) was added dropwise to a solution of l-bromo-3-cyanobenzene (936 mg, 5.17 mmol) in THF (10 mL) at 0° C. under N.sub.2. After stirring at 0° C. for 2 h, a solution of E-2761(200 mg, 0.517 mmol) in THF (10 mL) was added at 0° C. under N.sub.2. The mixture was stirred at 0° C. for 2 h and quenched with saturated aqueous NH.sub.4Cl (30 mL) was added. The aqueous phase was extracted with EtOAc (3×20 mL). The combined organic phase was washed with saturated brine (2×20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give an oil. The mixture was purified by flash column (0-20% of EtOAc in PE) to give DA-62-5_1 (140 mg, 55%) as a solid.

[1293] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.70-7.65 (m, 1H), 7.60-7.50 (m, 2H), 7.50-7.40 (m, 1H), 5.30-5.28 (m, 1H), 4.72-4.52 (m, 1H), 2.40-2.30 (m, 1H), 2.05-1.50 (m, 5H), 1.50-1.30 (m, 9H), 1.SO-LIS (m, 4H), 1.15-0.88 (m, 15H), 0.88-0.78 (m, 4H), 0.66 (s, 3H).

Synthesis of 6997 and 6998

[1294] ##STR00369##

[1295] DA-62-5_1 (140 mg, 0.285 mmol) was purified by SFC (Column: AD (150×4.6 mm, 3 um), Gradient: 5%-40% B (A: CO.sub.2 B: ethanol) Flow rate: 2.5 mL/min) to afford DA-62-5(30.0 mg, 22%) as a solid and DA-62-11 (38.0 mg, 27%) as a solid.

[1296] 6997:

[1297] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 1H), 7.10-7.02 (m, 2H), 7.00-6.90 (m, 1H), 5.30-5.25 (m, 1H), 4.68-4.55 (m, 1H), 2.40-2.30 (m, 1H), 2.05-1.90 (m, 3H), 1.90-1.50 (m, 7H), 1.50-1.30 (m, 8H), 1.30-0.86 (m, 16H), 0.86-0.76 (m, 3H), 0.66 (s, 3H).

[1298] LCMS Rt=1.202 min in 2.0 min chromatography, 30-90 AB, purity 99%, MS ESI calcd. for C.sub.33H.sub.46NO [M+H−H.sub.2O].sup.+ 472, found 472.

[1299] 6998:

[1300] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 1H), 7.10-7.02 (m, 2H), 7.00-6.90 (m, 1H), 5.30-5.25 (m, 1H), 4.65-4.53 (m, 1H), 2.40-2.30 (m, 1H), 2.05-1.90 (m, 4H), 1.90-1.50 (m, 7H), 1.50-1.30 (m, 7H), 1.30-0.86 (m, 16H), 0.86-0.80 (m, 3H), 0.66 (s, 3H).

[1301] LCMS Rt=1.194 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.33H.sub.46NO [M+H−H.sub.2O].sup.+ 472, found 472.

Example 70: Synthesis of 7030 and 7032

[1302] ##STR00370##

[1303] Stereochemistry assigned based on NMR data.

[1304] The experimental of intermediate DA-023-3 can be found in Example 126.

Synthesis of DA-023

[1305] ##STR00371##

[1306] Isopropylmagnesium chloride (1.29 mL, 2 M in THF, 2.58 mmol) was added to a suspension of 2-bromopyridine (407 mg, 2.58 mmol) in THF (4 mL) at 0° C. The mixture was stirred at 25° C. for 2 h. 5.35 mL of the freshly prepared pyridin-2-ylmagnesium bromide (5.35 mL, ca. 0.48 M in THF, 2.57 mmol) was added to a solution of DA-023-3 (200 mg, 514 umol) in THF (5 mL) at 0° C. The mixture was stirred at 25° C. for 16 hrs. The mixture was poured into water (20 mL) and extracted with EtOAc (2×30 mL). The combined organic layer was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (PE/EtOAc=5/1 to 3/1) to give DA-023 (150 mg, 63%) as a solid.

Synthesis of DA-023-15, 16

[1307] ##STR00372##

[1308] DA-023 (150 mg) was purified by SFC (Column: AS (250 mm*30 mm, 5 um), Condition: 0.1% NH.sub.3H.sub.2O IPA, Gradient: from 25% to 25%, Flow Rate (ml/min): 50 mL/min, 25° C.) to afford DA-023-15 (31 mg, 21%) and DA-023-16 (55 mg, 37%) as a solid.

[1309] 7030:

[1310] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.53 (d, J=4 Hz, 1H), 7.70-7.65 (m, 1H), 7.25-7.24 (m, 1H), 7.20-7.17 (m, 1H), 4.71-4.68 (m, 1H), 1.95-1.90 (m, 1H), 1.84-1.51 (m, 12H), 1.47-1.13 (m, 11H), 1.12-0.81 (m, 16H), 0.63-0.58 (m, 4H).

[1311] LCMS Rt=0.923 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.31H.sub.50NO.sub.2 [M+H].sup.+ 468, found 468.

[1312] 7032:

[1313] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.53 (d, J=4 Hz, 1H), 7.70-7.65 (m, 1H), 7.25-7.24 (m, 1H), 7.20-7.17 (m, 1H), 4.71-4.68 (m, 1H), 1.95-1.83 (m, 2H), 1.81-1.31 (m, 16H), 1.28-1.03 (m, 9H), 1.00-0.81 (m, 13H), 0.63-0.58 (m, 4H).

[1314] LCMS Rt=0.914 min in 2.0 min chromatography, 30-90 AB, purity 99.5%, MS ESI calcd. For C.sub.31H.sub.50NO.sub.2 [M+H].sup.+ 468, found 468.

Example 71: Synthesis of 7147 and 7146

[1315] ##STR00373##

[1316] The experimental of intermediate E-02761 can be found in Example 63.

Synthesis of DA-62-3_1

[1317] ##STR00374##

[1318] 1-bromo-4-fluorobenzene (900 mg, 5.14 mmol) was added to a suspension of magnesium (124 mg, 5.14 mmol) and a small amount of iodine (130 mg, 0.514 mmol) in tetrahydrofuran (10 mL). The mixture was stirred for 2 h at 50° C. A solution of E-2761 (200 mg, 0.517 mmol) in THF (10 mL) was added to the Grignard mixture at 15° C. under N.sub.2. The mixture was stirred at 15° C. for 2 h and quenched with saturated aqueous NH.sub.4Cl(30 mL). The aqueous phase was extracted with EtOAc (3×20 mL). The combined organic phase was washed with saturated brine (2×20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give an oil, which was purified by flash column (0-20% of EtOAc in PE) to give DA-62-3_1 (120 mg, 48%) as a solid.

[1319] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 2H), 7.08-6.95 (m, 2H), 5.28 (brs, 1H), 4.63-4.53 (m, 1H), 2.38-2.30 (m, 1H), 2.00-1.50 (m, 10H), 1.50-1.28 (m, 8H), 1.28-1.00 (m, 10H), 1.00-0.80 (m, 9H), 0.66 (s, 3H).

[1320] LCMS Rt=1.453 min in 2.0 min chromatography, 30-90 AB, purity 84%, MS ESI calcd. for C.sub.32H.sub.44F [M+H−2H.sub.2O].sup.+ 447, found 447.

Synthesis of 7146 and 7147

[1321] ##STR00375##

[1322] DA-62-3_1 (120 mg, 248 umol) was purified by SFC (Column: AD (150×4.6 mm, 3 um), Gradient: 5%-40% B (A: CO.sub.2 B: ethanol) Flow rate: 2.5 mL/min) to afford 7146 (30 mg, 25%) as a solid and 7147 (27 mg, 23%) as a solid.

[1323] 7146

[1324] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 2H), 7.08-6.95 (m, 2H), 5.28 (brs, 1H), 4.63-4.53 (m, 1H), 2.38-2.30 (m, 1H), 2.05-1.85 (m, 3H), 1.75-1.50 (m, 7H), 1.50-1.28 (m, 7H), 1.28-0.86 (m, 15H), 0.85-0.80 (m, 3H), 0.66 (s, 3H).

[1325] LCMS Rt=1.434 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.44F [M+H−2H.sub.2O].sup.+ 447, found 447.

[1326] 7147

[1327] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.25 (m, 1H), 7.10-7.02 (m, 2H), 7.00-6.90 (m, 1H), 5.30-5.25 (m, 1H), 4.65-4.53 (m, 1H), 2.40-2.30 (m, 1H), 2.05-1.90 (m, 3H), 1.90-1.50 (m, 7H), 1.50-1.28 (m, 8H), 1.28-0.86 (m, 16H), 0.86-0.80 (m, 3H), 0.66 (s, 3H).

[1328] LCMS Rt=1.437 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.44F [M+H−2H.sub.2O].sup.+ 447, found 447.

Synthesis of 7146 to Determine Stereochemistry

[1329] ##STR00376##

[1330] To a solution of Me.sub.3SI (32.8 g, 161 mmol) in DMSO (100 mL) and THF (50 mL) was added NaH (6.43 g, 60%, 161 mmol) at 25° C. The reaction mixture was stirred for 20 mins at 25° C., then was cooled to 0° C. and added 4-fluorobenzaldehyde (10 g, 80.5 mmol) in THF (50 mL). The reaction mixture was stirred for 1 h, treated with water (200 mL) and extracted with EtOAc (2×200 mL). The organic phase was washed with water (2×200 mL), brine (200 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude product, which was purified by combi flash (EtOAc/PE=0-5%) to give ST-200-095-005_2P (5.5 g, impure) as colorless oil, which was purified by combi flash (EtOAc/PE=0-1%) to give ST-200-095-005_2P (4.0 g, 73%) as an oil.

[1331] LCMS Rt=1.192 min in 7 min chromatography, 30-90CD_7MIN_E, purity 99%, MS ESI calcd.

[1332] SFC Peak 1: Rt=2.209 min and Peak 2 Rt=2.407 min in 10 min chromatography, O1_EtOH_DEA_5_40_25ML (“Column: (S,S)Whelk-O1 250*4.6 mm, 5 um, Mobile phase: A: CO.sub.2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.”).

[1333] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.20-7.16 (m, 2H), 6.99-6.94 (m, 2H), 3.79-3.76 (m, 1H), 3.08-3.05 (m, 1H), 2.71-2.68 (m, 1H).

[1334] To a solution of R,R-cat (69.4 mg, 0.115 mmol) in toluene (5 mL) was added AcOH (70.8 mg, 1.18 mmol). The mixture was stirred at 25° C. open to air for 30 min and concentrated in vacuo to leave a crude solid. The resulting catalyst residue was dissolved in ST-200-095-005_2P (4 g, 28.9 mmol) at 25° C. The reaction flask was cooled to 0° C., followed by adding H.sub.2O (284 mg, 15.8 mmol) dropwise over 5 min. The reaction was allowed to warm to 25° C. and stirred for 16 hrs. The reaction mixture was purified directly by silica gel chromatography (PE %=100%) to afford ST-200-095-005_3P (460 mg, 11%) as an oil. The ee % was 97%.

[1335] LCMS Rt=1.605 min in 2 min chromatography, 10-80CD_3MIN_E, purity 94%, DAD1 A, Sig=220.

[1336] SFC Peak 1: Rt=2.202 min and Peak 2 Rt=2.398 min in 10 min chromatography, O1_EtOH_DEA_5_40_25ML (“Column: (S,S)Whelk-O1 250*4.6 mm, 5 um, Mobile phase: A: CO.sub.2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.”).

[1337] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.20-7.16 (m, 2H), 6.99-6.94 (m, 2H), 3.79-3.76 (m, 1H), 3.08-3.05 (m, 1H), 2.71-2.68 (m, 1H).

[1338] To a THF (5 mL) under N.sub.2 at −70° C. was added n-BuLi (2.5 M, 4.12 mmol, 1.64 mL). After that, a suspension of ST-200-550_15-1 (800 mg, 1.65 mmol) in THF (5 mL) was added drop-wise to give a suspension. After stirring at −70° C. for 30 min, a solution of ST-200-095-005_3P (455 mg, 3.30 mmol) was added. Then reaction was stirred at stirred at 25° C. for 16 hours. The mixture was poured into ice-water (20 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to afford ST-200-095-005_4P (1 g, crude) as an oil, which was used directly for the next step.

[1339] To a solution of ST-200-095-005_4P (1 g, crude) in MeOH (50 mL) was added NiCl.sub.2 (5 mg) and Mg powder (1.533 g, 64.0 mmol) at 65° C. in four portions. The reaction mixture was cooled to 20° C. and quenched by HCl (1M, 100 mL). The mixture was extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the crude, which was purified by flash column (0˜5% of EtOAc in PE) to give ST-200-095-005_5P (41 mg, 5%) as a solid.

[1340] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.35-7.27 (m, 2H), 7.05-6.98 (m, 2H), 5.35-5.31 (m, 1H), 4.62-4.58 (m, 1H), 2.41-2.31 (m, 1H), 2.10-1.90 (m, 4H), 1.75-1.58 (m, 8H), 1.52-1.05 (m, 14H), 1.02 (s, 3H), 1.00-0.76 (m, 8H), 0.66 (s, 3H).

[1341] LCMS Rt=1.272 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.32H.sub.44F [M+H−2H.sub.2O].sup.+ 447, found 447.

[1342] SFC Peak 1: Rt=2.124 min in 10 min chromatography, AD_3_EtOH_DEA_40_25ML (“Column: (S,S)Whelk-O1 250*4.6 mm, 5 um, Mobile phase: A: CO.sub.2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.”).

Example 72: Synthesis of 7281 and 7282

[1343] ##STR00377##

[1344] The experimental of intermediate E-2761 can be found in Example 63.

Synthesis of DA-62-1_1

[1345] ##STR00378##

[1346] n-BuLi(2.5 M, 2.05 mL) was added dropwise to a solution of l-bromo-2-fluorobenz(900 mg, 5.14 mmol) in THF (10 mL) at −78° C. under N.sub.2. After stirring at −78° C. for 30 min, a solution of E-2761 (200 mg, 0.517 mmol) in THF (10 mL) was added at −78° C. under N.sub.2. The mixture was stirred at −78° C. for 30 min and quenched with saturated aqueous NH.sub.4Cl(30 mL). The aqueous phase was extracted with EtOAc (3×20 mL). The combined organic phase was washed with saturated brine (2×20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give an oil, which was purified by flash column (0-20% of EtOAc in PE) to give DA-62-1_1 (110 mg, 44%) as a solid.

[1347] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.50-7.42 (m, 1H), 7.25-7.10 (m, 2H), 7.10-6.96 (m, 1H), 5.32-5.25 (m, 1H), 5.02-4.92 (m, 1H), 2.40-2.30 (m, 1H), 2.05-1.50 (m, 9H), 1.50-1.32 (m, 8H), 1.32-1.18 (m, 3H), 1.18-0.75 (m, 17H), 0.66 (s, 3H).

[1348] LCMS Rt=1.380 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.44F [M+H−2H.sub.2O].sup.+ 447, found 447.

Synthesis of 7281 and 7282

[1349] ##STR00379##

[1350] DA-62-11 (110 mg, 227 umol) was purified by SFC (Column: AD (150×4.6 mm, 3 um), Gradient: 5%-40% B (A: CO.sub.2 B: ethanol) Flow rate: 2.5 mL/min) to afford DA-62-1 (12 mg, 11%) as a solid and DA-62-7 (30 mg, impure) as a solid. The impure DA-62-7 (30 mg, impure) was purified by SFC (Column: AD (150×4.6 mm, 3 um), Gradient: 5%-40% B (A: CO.sub.2 B: ethanol) Flow rate: 2.5 mL/min) to afford DA-62-7(6 mg, 6%) as a solid.

[1351] 7281

[1352] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.48-7.42 (m, 1H), 7.26-7.10 (m, 2H), 7.06-6.96 (m, 1H), 5.32-5.25 (m, 1H), 5.02-4.92 (m, 1H), 2.40-2.30 (m, 1H), 2.05-1.50 (m, 11H), 1.50-1.30 (m, 7H), 1.30-1.18 (m, 2H), 1.18-0.75 (m, 17H), 0.66 (s, 3H).

[1353] LCMS Rt=1.362 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.44F [M+H−2H.sub.2O].sup.+ 447, found 447.

[1354] 7282

[1355] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.48-7.42 (m, 1H), 7.26-7.10 (m, 2H), 7.06-6.96 (m, 1H), 5.32-5.25 (m, 1H), 5.02-4.92 (m, 1H), 2.40-2.30 (m, 1H), 2.05-1.50 (m, 10H), 1.50-1.32 (m, 8H), 1.32-0.86 (m, 16H), 0.86-0.78 (m, 3H), 0.66 (s, 3H).

[1356] HPLC Rt=5.32 min in 7 min chromatography, 50-100 AB, purity 98%,

[1357] MS ESI calcd. for C.sub.32H.sub.44F [M+H−2H.sub.2O].sup.+ 447, found 447.

Synthesis to Determine Stereochemistry

[1358] ##STR00380##

[1359] To a solution of Me.sub.3SOI (123 g, 562 mmol) in DMSO (150 mL) and THF (75 mL) was added t-BuOK (63 g, 562 mmol) in portions at 25° C. The mixture was stirred at 40° C. for 30 min. Then ST-200-095-005_1 (35 g, 281 mmol) in 75 ml of THF was added dropwise to the mixture at 0° C. After stirring at 25° C. for 1 h, the mixture was poured into ice-water (100 mL) and extracted with EtOAc (2×50 mL). The combined organic phase was washed with brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated in vacuum. The residue was purified by silica gel column eluted with (PE/EtOAc=20/1) to afford ST-200-095-005_2 (24 g, 62%) as an oil.

[1360] .sup.1HNMR (400 MHz, CDCl3) δ 7.22-7.16 (m, 1H), 7.13-7.02 (m, 2H), 6.99-6.95 (m, 1H), 4.07 (t, J=4 Hz, 1H), 3.11-3.08 (m, 1H), 2.71-2.70 (m, 1H).

[1361] To a solution of R, R-cat (86.9 mg, 0.144 mmol) in toluene (5 mL) was added AcOH (86.4 mg, 1.44 mmol). The mixture was stirred at 25° C. for 30 mins. The solution was concentrated in vacuum to give a crude solid. The resulting catalyst residue was dissolved in ST-200-095-005_2 (5 g, 36.1 mmol) at 25° C., the reaction mixture was cooled to 0° C., and water (356 mg, 19.8 mmol) was added dropwise. The mixture was warmed to 25° C. and stirred for 16 hrs. The product was purified by silica gel column (PE/EtOAc=12/l to 8/1) to give ST-200-095-005_3 (1 g, 20%) as an oil.

[1362] .sup.1HNMR (400 MHz, CDCl3) δ 7.22-7.16 (m, 1H), 7.13-7.08 (m, 1H), 7.06-7.02 (m, 1H), 6.99-6.95 (m, 1H), 4.07 (t, J=4 Hz, 1H), 3.11-3.08 (m, 1H), 2.71-2.70 (m, 1H).

[1363] SFC Peak 1: Rt=2.015 min in 10 min chromatography, SS Whelk O1_EtOH_DEA_5_40_25ML (“Column: (S,S)Whelk-O1 250*4.6 mm, 5 um, Mobile phase: A: CO2 B: ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.”), 97.1% ee. To a THF (2 mL) under N.sub.2 at −70° C. was added n-BuLi (2.5 M, 1.54 mmol, 0.616 mL). After that, a suspension of ST-200-095-005_3 (300 mg, 0.618 mmol) in THF (2 mL) was added drop-wise to give a suspension. After stirring at −70° C. for 30 min, a solution of (2R)-2-(2-fluorophenyl)oxirane (127 mg, 0.926 mmol) was added. The reaction was stirred at stirred at 25° C. for 16 hours. The mixture was poured into ice-water (20 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to afford ST-200-095-005_4 (350 mg, crude) as a solid, which was used directly for the next step.

[1364] To a solution of ST-200-095-005_4 (350 mg, 0.561 mmol) and nickel (II) chloride (7 mg, 0.056 mmol) in MeOH (30 mL) was added Mg (294 mg, 11.2 mmol) was added at 25° C. The mixture was stirred at 50° C. for 1 h. After cooling, the mixture was quenched with HCl (100 mL, 2M) until the reaction became clear and extracted with EtOAc (2×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered concentrated and purified by a silica gel column (PE/EtOAc=10/1 to 3/1) to give ST-200-095-006 (150 mg, 56%) as a solid.

[1365] .sup.1HNMR (400 MHz, CDCl3) 7.44 (t, J=8 Hz, 1H), 7.25-7.21 (m, 1H), 7.15 (t, J=8 Hz, 1H), 7.02 (t, J=8 Hz, 1H), 5.28-5.26 (m, 1H), 5.00-4.90 (m, 1H), 2.39-2.32 (m, 1H), 2.04-1.86 (m, 4H), 1.84-1.60 (m, 7H), 1.55-1.32 (m, 9H), 1.29-0.99 (m, 10H), 0.96-0.90 (m, 4H), 0.84 (t, J=8 Hz, 3H), 0.66 (s, 3H).

[1366] LCMS Rt=1.290 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.32H.sub.44F [M−2H.sub.2O+H.sup.+]=447, found 447.

[1367] SFC Rt=5.494 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML (“Column: Chiralpak AD-3 150×4.6 mm I.D., 3 um Mobile phase: A: CO2B: ethanol (0.05% DEA) Gradient: from 5% to 40% of B in 5 min and hold 40% for 2.5 min, then 5% of B for 2.5 min Flow rate: 2.5 mL/min Column temp.: 35° C.”), 98.86% de.

Example 73: Synthesis of 7300 and 7399

[1368] ##STR00381##

[1369] The stereochemistry has been randomly assigned. The experimental of intermediate E-2761 can be found in Example 63.

Synthesis of DA-62-6_1

[1370] ##STR00382##

[1371] n-BuLi (2.5 M, 3.09 mL) was added dropwise to a solution of 2-bromobenzonitrile (1.41 g, 7.75 mmol) in THF (10 mL) at −78° C. under N.sub.2. The mixture was stirred at −78° C. for 30 min. E-2761 (600 mg, 1.55 mmol) in THF (10 mL) was added at −78° C. under N.sub.2. The mixture was stirred at −78° C. for 30 min and quenched with aqueous NH.sub.4Cl (30 mL). The aqueous phase was extracted with EtOAc (3×50 mL). The combined organic phase was washed with saturated brine (2×50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give an oil, which was purified by flash column (0-20% of EtOAc in PE) to give DA-62-6_1 (300 mg, impure) as a solid.

[1372] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.90-7.80 (m, 1H), 7.60-7.40 (m, 2H), 7.40-7.26 (m, 1H), 5.50-5.40 (m, 1H), 5.30 (brs, 1H), 2.42-2.30 (m, 1H), 2.10-1.50 (m, 9H), 1.60-1.30 (m, 4H), 1.30-0.90 (m, 20H), 0.90-0.75 (m, 4H), 0.66 (s, 3H).

Synthesis of 7399 and 7300

[1373] ##STR00383##

[1374] DA-62-6_1 (300 mg, 0.612 mmol) was purified by SFC (Column: AD (150×4.6 mm, 3 um), Gradient: 5%-40% B (A: CO.sub.2 B: ethanol) Flow rate: 2.5 mL/min) to afford DA-62-6 (30.0 mg, impure) as a solid and DA-62-12 (80 mg, impure) as solid. Impure DA-62-6 (30 mg, 0.0612 mmol) was purified by SFC (Column: AS (250 mm*30 mm, 5 um), Gradient: 25%-25% B (A: CO.sub.2 B: 0.1% NH.sub.3H.sub.2O ETOH) Flow rate: 60 mL/min) to afford DA-62-6 (4 mg, 13%) as solid. Impure DA-62-12 (80 mg, 0.163) was purified by SFC (Column: AS (250 mm*30 mm, 5 um), Gradient: 25%-25% B (A: CO.sub.2 B: 0.1% NH.sub.3H.sub.2O ETOH) Flow rate: 60 mL/min) to afford an impure solid, which was triturated from H.sub.2O (10 mL) at 90° C. to give DA-62-12 (3 mg, 4%) as a solid.

[1375] 7399:

[1376] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.88-7.80 (m, 1H), 7.58-7.42 (m, 2H), 7.35-7.30 (m, 1H), 5.45-5.36 (m, 1H), 5.30-5.27 (m, 1H), 2.40-2.30 (m, 1H), 2.10-1.90 (m, 4H), 1.85-1.50 (m, 8H), 1.50-1.30 (m, 5H), 1.30-1.88 (m, 17H), 0.86-0.76 (m, 3H), 0.66 (s, 3H).

[1377] LCMS Rt=0.908 min in 2.0 min chromatography, 30-90 AB, purity 95%, MS ESI calcd. for C.sub.33H.sub.48NO.sub.2 [M+H].sup.+ 490, found 490.

[1378] 7300:

[1379] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.88-7.75 (m, 1H), 7.58-7.40 (m, 2H), 7.30-7.26 (m, 1H), 5.50-5.40 (m, 1H), 5.30-5.25 (m, 1H), 2.40-2.37 (m, 1H), 2.10-1.50 (m, 13H), 1.50-1.10 (m, 14H), 1.10-0.76 (m, 10H), 0.66 (s, 3H).

[1380] LCMS Rt=0.923 min in 2.0 min chromatography, 30-90 AB, purity 94%, MS ESI calcd. for C.sub.33H.sub.48NO.sub.2 [M+H].sup.+ 490, found 490.

Example 74: Synthesis of 7467 and 7468

[1381] ##STR00384##

[1382] The stereochemistry was randomly assigned. The experimental of intermediate ST-200-081-001_4 could be found in Example 127.

Synthesis of ST-200-081-001_5

[1383] ##STR00385##

[1384] To a solution of ST-200-081-001_4 (1.3 g, 3.01 mmol) in THF (10 mL) was added PhLi (7.5 mL, 2 M in ether, 15 mmol) at 0° C. under N.sub.2 and the mixture was stirred at 25° C. for 30 minutes. After quenching with saturated NH.sub.4Cl (40 mL), the mixture was extracted with EtOAc (3×20 mL). The combined organic layer was washed with brine dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum to give ST-200-081-001_5 (1.6 g, crude) as a solid, which was used directly without further purification.

[1385] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.01-7.93 (m, 2H), 7.63-7.29 (m, 7H), 5.34-5.27 (m, 1H), 3.08-2.83 (m, 2H), 2.47-2.38 (m, 1H), 2.01-1.67 (m, 7H), 1.52-1.14 (m, 11H), 1.11 (s, 3H), 1.03-0.97 (m, 7H), 0.69 (s, 3H).

Synthesis of ST-200-081-001_6

[1386] ##STR00386##

[1387] To a solution of ST-200-081-001_5 (1.6 g, 3.56 mmol) in THF (10 mL) was added MeLi (11.1 mL, 2 M in ether, 17.8 mmol) at 0° C. under N.sub.2 and the mixture was stirred at 25° C. for 30 minutes. The reaction mixture was quenched by saturated NH.sub.4Cl (10 mL) and extracted with ethyl acetate (3×20 mL). The organic layer was washed with brine (60 mL), dried over Na.sub.2SO.sub.4 and filtered, concentrated in vacuum and purified by flash column (0-30% of EtOAc in PE) to give ST-200-081-001_6 (1 g, 57%) as a solid.

[1388] LCMS Rt=1.374 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.32H.sub.45 [M+H−2H.sub.2O].sup.+ 429, found 429.

Synthesis of 7467 and 7468

[1389] ##STR00387##

[1390] ST-200-081-001_6 (1 g, 2.15 mmol) was purified by SFC (column: AD (250 mm*30 mm, 5 um)), gradient: 40-40% B (A=0.1% NH.sub.3/H.sub.2O, B=EtOH), flow rate: 60 mL/min) to give impure ST-200-081-001 (Peak 1, 390 mg, 39%) and impure ST-200-081-002 (Peak 2, 220 mg, 22%) as a solid. To a solution of impure ST-200-081-001 (390 mg) in THF (15 mL) was added Pd(OH).sub.2/C (wet, 300 mg) and the mixture was degassed and back-filled with H.sub.2 for 3 times. After that, the reaction was stirred at 15° C. under 15 psi of H.sub.2 for 4 h. The reaction mixture was filtered through a pad of celite washed with THF (100 mL). The filtrate was concentrated and purified by SFC (column: AD (250 mm*30 mm, 5 um)), gradient: 40-40% B (A=0.1% NH.sub.3/H.sub.2O, B=EtOH), flow rate: 60 mL/min) to give ST-200-081-001 (270 mg, 71%) as a solid. Impure ST-200-081-002 was triturated in boiling MeCN (200 mL) concentrated in vacuum to give ST-200-081-002 (208 mg, 94%) as a solid.

[1391] ST-200-081-001 (7468):

[1392] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.46-7.39 (m, 2H), 7.37-7.29 (m, 2H), 7.26-7.21 (m, 1H), 5.33-5.26 (m, 1H), 2.46-2.37 (m, 1H), 2.01-1.64 (m, 9H), 1.56-1.53 (m, 5H), 1.52-1.12 (m, 10H), 1.10 (s, 3H), 1.09-1.01 (m, 3H), 0.99 (s, 3H), 0.97-0.86 (m, 5H), 0.62 (s, 3H).

[1393] LCMS Rt=1.359 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.32H.sub.45 [M+H−2H.sub.2O].sup.+ 429, found 429.

[1394] SFC Rt=5.916 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100% de.

[1395] ST-200-081-002 (6467):

[1396] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.45-7.39 (m, 2H), 7.36-7.31 (m, 2H), 7.25-7.19 (m, 1H), 5.33-5.26 (m, 1H), 2.47-2.37 (m, 1H), 2.01-1.57 (m, 10H), 1.55-1.31 (m, 12H), 1.21-1.12 (m, 2H), 1.10 (s, 3H), 1.08-1.02 (m, 2H), 0.99 (s, 3H), 0.98-0.85 (m, 6H), 0.63 (s, 3H).

[1397] LCMS Rt=1.367 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.32H.sub.45 [M+H−2H.sub.2O].sup.+ 429, found 429.

[1398] SFC Rt=6.397 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 97.26% de.

Example 74B. Biological Data

[1399] The Experiments were conducted as described in Example 2 and the results are reported in Table 2-66.

TABLE-US-00004 TABLE 2-66 Avg Avg Emax Avg Emax Avg EC50 2A EC50 2B Compound 2A (nM) (%) 2B (nM) (%) [00388] 154 >10000 18.2 >10000 56.8 [00389] 356 414.6 59.6 923.4 156.3 [00390] 456 6710.0 53.4 6746.8 74.8 [00391] 559 1571.3 204.3 1564.3 373.2 [00392] 255 646.4 159.4 153.0 131.8 [00393] 761 475.9 301.6 249.1 197.3 [00394] 861 199.6 183.1 246.9 389.8 [00395] 961 616.7 138.5 179.8 97.4 [00396] 6347 268.3 64.7 310.7 131.9 [00397] 6348 259.61 79.65 663.21 132.91 [00398] 6457 731.9 198.3 523.6 168.1 [00399] 6458 >10000 30.9 928.9 79.4 [00400] 6459 1327.1 156.2 909.0 184.2 [00401] 6544 558.8 253.2 497.7 414.6 [00402] 6754 >10000 22.3 >10000 38.8 [00403] 6755 >10000 22.8 >10000 44.2 [00404] 6571 189.8 157.5 266.6 241.0 [00405] 6572 378.5 128.9 1341.5 250.6 [00406] 6680 1370.7 175.4 464.4 123.4 [00407] 6681 365.3 171.6 359.9 165.3 [00408] 6895 174.1 240.9 328.8 354.3 [00409] 6896 518.9 215.9 1676.0 402.2 [00410] 6997 373.4 327.2 414.3 314.7 [00411] 6998 357.6 337.9 304.3 303.4 [00412] 7030 >10000 28.3 >10000 74.7 [00413] 7032 >10000 28.3 874.3 63.8 [00414] 7146 119.8 103.7 215.9 167.4 [00415] 7147 147.0 200.5 91.7 241.5 [00416] 7281 248.37 98.1 302.7 125.7 [00417] 7282 95.9 208.3 173.7 289.7 [00418] 7399 696.1 150.1 1581.2 179.5 [00419] 7300 >10000 37.5 >10000 46.9 [00420] 7467 >10000 20.5 >10000 8.8 [00421] 7468 61.4 273.4 54.2 317.4 [00422] 660 700.6 195.2 516.8 281.1 [00423] 6010 1104.9 238.7 1573.5 184.0 [00424] 6051 500.2 292.7 1262.1 292.4 [00425] 6052 425.9 101.8 232.3 92.2

Example 75: Syntheses of Compounds 175, 1A75, and 1B75

[1400] ##STR00426## ##STR00427## ##STR00428##

Step 1

[1401] To freshly prepared liquid ammonia (1.0 L) was added lithium (12.7 g, 1.82 mol) in portions at −70° C. The mixture became deep blue. After stirring at −70° C. for 1 h, a solution of A175 (50 g, 183 mmol) and t-butanol (26.9 g, 364 mmol) in dry THF (600 mL) was added to this mixture with strong stirring, and the temperature was maintained below −60° C. The resultant mixture was stirred at −70° C. for 2 hrs. Ammonium chloride (150 g) was added to reaction mixture. The mixture was warmed to 25° C. and stirred for 16 hrs. The reaction mixture was neutralized with aq. HCl (2.5 M, 1000 mL) and filtered. The filtrate was extracted with EtOAc (1 L×2), washed with brine (1 L), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford A2 (45 g, crude) as a solid.

[1402] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.65-3.57 (m, 1H), 2.06-1.66 (m, 5H), 1.43-0.75 (m, 16H), 0.74 (s, 3H), 0.73-0.59 (m, 3H).

Step 2

[1403] To a solution of A275 (43 g, 155 mmol) in CH.sub.2Cl.sub.2 (600 mL) was added silica gel (75 g, w/w=1/1.5) and pyridinium chlorochromate (52.3 g, 243 mmol) at 25° C. The mixture was stirred at 25° C. for 2 hrs. The mixture was filtered and the filtrate was concentrated in vacuum. The residue was purified by column chromatography on silica gel (PE/EtOAc=20/l to 5/1) to afford A375 (22.0 g, 50%) as a solid.

[1404] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.52-2.38 (m, 2H), 2.38-2.28 (m, 3H), 2.15-2.05 (m, 2H), 2.05-1.65 (m, 5H), 1.55-1.40 (m, 2H), 1.40-1.15 (m, 6H), 1.15-0.92 (m, 1H), 0.90 (s, 3H), 0.89-0.80 (m, 1H), 0.80-0.65 (m, 1H).

Step 3

[1405] To a solution of BHT (48 g, 218 mmol) in toluene (120 mL) was added AlMe.sub.3 (2 M in toluene, 120 mL, 218 mmol) at 0° C. and stirred at 10° C. for 1 h. To the MAD solution (109 mmol in 120 mL toluene) was added a solution of A375 (10 g, 36.4 mmol) in DCM (30 mL) at −78° C. After stirring at −78° C. for 1 h, MeMgBr (36.3 mL, 109 mmol) was added at −78° C. The mixture was stirred at −78° C. for 20 mins. The reaction mixture was treated with saturated citric acid (50 mL). The organic phase was separated and the aqueous phase extracted with EtOAc (80 mL). The organic phase was washed with brine (100 mL), dried over Na.sub.2SO.sub.4, concentrated in vacuum to give a crude product, which was purified by flash column (0-30% of EtOAc in PE) to give A476 (6 g, 57%) as a solid.

[1406] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.50-2.45 (m, 1H), 2.13-1.96 (m, 1H), 1.95-1.70 (m, 6H), 1.70-1.60 (m, 2H), 1.58-1.45 (m, 1H), 1.45-0.95 (m, 13H), 0.95-0.83 (m, 4H), 0.80-0.65 (m, 2H).

Step 4

[1407] To a suspension of PPh.sub.3EtBr (22.9 g, 61.8 mmol) in THF (30 mL) was added t-BuOK (6.93 g, 61.8 mmol) at 20° C. After stirring at 40° C. for 30 min, a solution of A475 (6 g, 20.6 mmol) in THF (20 mL) was added at 40° C. and the reaction mixture was stirred at 40° C. for 1 h. The reaction mixture was poured into 50 g of crushed ice and stirred for 15 minutes. The organic layer was separated and the water phase was extracted with EtOAc (30 mL). The combined organic phase was concentrated in vacuo to give a thick oil. The residue was dissolved in 90 mL of MeOH at 60° C., following by treating with 90 mL of water. A precipitate formed. After stirring at 60° C. for 1 h, the precipitate was collected by filtration and washed with a solution of MeOH/water (15 mL/15 mL) and dried under vacuum to give the product A575 as a solid.

[1408] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.18-5.06 (m, 1H), 2.42-2.29 (m, 1H), 2.29-2.10 (m, 2H), 1.95-1.84 (m, 1H), 1.84-1.46 (m, 10H), 1.46-1.25 (m, 2H), 1.25-1.05 (m, 10H), 1.05-0.80 (m, 5H), 0.75-0.60 (m, 2H).

Step 5

[1409] To a solution of A575 (4.8 g, 3.30 mmol) and methyl propiolate (4.21 mL, 47.4 mmol) in anhydrous dichloromethane (100 mL) under N.sub.2 at 0° C. was added dropwise diethylaluminum chloride (1.0 M in toluene, 63.2 mL, 63.2 mmol). The mixture was stirred at 20° C. for 16 hours. The reaction mixture was quenched with aqueous citric acid (100 mL) at 0° C. carefully, and the inner temperature was maintained below 10° C. The resultant mixture was filtered through a pad of celite and extracted with DCM (2×200 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated to give crude A675 as a solid. Combined with another batch, the crude product was purified by flash column (0-30% of EtOAc in PE) to give 6 g of impure product as a solid. The solid was recrystallized from DCM/PE (20 mL/80 mL) to give 3.1 g of pure product as a solid. The mother liquid was concentrated to give 2.8 g of the product as a solid. A total of 5.9 g of the product was obtained (79% yield).

[1410] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.96 (dd, J=8.0, 15.6 Hz, 1H), 5.83 (dd, J=0.8, 15.6 Hz, 1H), 5.45-5.38 (m, 1H), 3.75 (s, 3H), 3.10-2.96 (m, 1H), 2.10-2.03 (m, 1H), 1.95-1.55 (m, 9H), 1.45-0.85 (m, 18H), 0.80-0.60 (m, 3H).

Step 6

[1411] To a solution of A675 (3 g, 7.76 mmol) in EtOAc (100 mL) was added Pd/C (100 mg, 10%, wet) and the mixture was degassed and back-filled with H.sub.2 for 3 times. The reaction was stirred at 15° C. under 15 psi of H.sub.2 for 16 hrs. The reaction mixture was filtered through a pad of celite washed with EtOAc (20 mL). The filtrate was concentrated to give 3 g of A775 as an oil, which was used directly for the next step.

[1412] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.66 (s, 3H), 2.43-2.30 (m, 1H), 2.30-2.15 (m, 1H), 1.95-1.50 (m, 10H), 1.50-0.96 (m, 17H), 0.96-0.80 (m, 5H), 0.75-0.050 (m, 5H).

Step 7

[1413] To a solution of A775 (3 g, 7.68 mmol) in THF (100 mL) was added LiAlH.sub.4 (580 mg, 15.3 mmol) at 0° C. under N.sub.2. After stirring at this temperature for 1 h, the reaction mixture was treated with water (2 mL) and then the pH was adjusted to 1-2 with saturated citric acid. The water phase was extracted with EtOAc (2×50 mL). The combined organic layers were washed with brine (2×100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to give 2.5 g of crude A875, which was used directly for the next step.

[1414] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.70-3.50 (m, 2H), 2.00-1.78 (m, 3H), 1.78-1.52 (m, 8H), 1.52-1.30 (m, 4H), 1.30-0.98 (m, 15H), 0.98-0.80 (m, 5H), 0.75-0.55 (m, 5H).

Step 8

[1415] To a solution of A875 (2.5 g, 6.89 mmol) in DCM (200 mL) was added PCC (2.94 g, 13.7 mmol) and silica gel (5 g). After stirring at 20° C. for 2 h, the reaction was filtered through a pad of celite washed with DCM (2×20 mL). The filtrate was concentrated to give 3 g of crude mixture, which was purified by flash column (0-30% of EtOAc in PE) to give 1.5 g of the product as a solid.

[1416] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.76 (t, J=1.2 Hz, 1H), 2.53-2.28 (m, 2H), 2.00-1.50 (m, 12H), 1.50-0.98 (m, 13H), 0.98-0.80 (m, 6H), 0.75-0.55 (m, 6H).

Step 9

[1417] To a solution of A975 (1.5 g, 4.16 mmol) in THF (100 mL) was added iPrMgCl (6.20 mL, 12.4 mmol) at 0° C. The reaction was stirred at this temperature for 1 h and quenched by adding water (50 mL) and saturated citric acid solution (100 mL). The mixture was then extracted with EtOAc (2×100 mL). The organic layers were combined and washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrate. The crude residue was purified by flash column (0-20% of EtOAc in DCM) to 1.4 g of the product as a solid. Compound 175 (1.3 g) was triturated with MeCN (50 mL) at 80° C. to give 175 (350 mg) as a solid.

[1418] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.38-3.25 (m, 1H), 1.98-1.92 (m, 1H), 1.92-1.76 (m, 2H), 1.76-1.50 (m, 9H), 1.50-1.40 (m, 4H), 1.40-0.97 (m, 14H), 0.97-0.80 (m, 12H), 0.73-0.53 (m, 5H).

[1419] LCMS Rt=1.309 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.27H.sub.45 [M+H−2H.sub.2O].sup.+ 369, found 369.

Step 10

[1420] To a solution of 175 (700 mg, 1.72 mmol) in pyridine (5 mL) was added BzCl (723 mg, 5.15 mmol) at 0° C. and the reaction was stirred at 20° C. for 18 h to give a solution. The reaction mixture was diluted with water (20 mL), extracted with EtOAc (2×20 mL). The organic layer was washed with brine (5×100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The crude residue was purified by flash column (0-10% of EtOAc in PE) to give 920 mg of an oil. The crude product was separated by SFC (AD (250 mm*30 mm, 5 um)), 0.1% NH.sub.3H.sub.2O-EtOH) to give 280 mg of peak 1 as A10-A75 as a solid and 285 mg of peak 2 as A10-B75 as a solid.

[1421] A10-A75:

[1422] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.07 (d, J=8.4 Hz, 2H), 8.01 (d, J=8.4 Hz, 2H), 7.58-7.48 (m, 2H), 7.48-7.36 (m, 4H), 5.00-4.90 (m, 1H), 2.40-2.30 (m, 1H), 2.30-2.20 (m, 1H), 2.00-1.85 (m, 3H), 1.80-1.30 (m, 13H), 1.30-0.88 (m, 22H), 0.88-0.70 (m, 1H), 0.70-0.50 (m, 4H).

[1423] A10-B75:

[1424] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.07 (d, J=8.4 Hz, 2H), 8.01 (d, J=8.4 Hz, 2H), 7.58-7.48 (m, 2H), 7.48-7.36 (m, 4H), 5.00-4.92 (m, 1H), 2.47-2.28 (m, 1H), 2.28-2.20 (m, 1H), 2.00-1.85 (m, 3H), 1.80-1.45 (m, 13H), 1.30-0.85 (m, 22H), 0.85-0.70 (m, 1H), 0.70-0.50 (m, 4H).

Step 11

[1425] To a solution of A10-A (280 mg, 0.46 mmol) in THF (5 mL) and MeOH (5 mL) was added NaOH (200 mg, 5.00 mmol) and H.sub.2O (2 mL) at 25° C. Then the solution was stirred at 50° C. for 16 h. The reaction solution was extracted with EtOAc (2×10 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give crude product which was purified by a silica gel column (PE/EtOAc=3:1) to give a solid, which was then triturated in hot MeCN (5 mL) to give the desired product Compound 1-A75 (102 mg, 55%) as a solid.

[1426] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.38-3.25 (m, 1H), 1.98-1.92 (m, 1H), 1.92-1.76 (m, 2H), 1.76-1.50 (m, 9H), 1.50-1.40 (m, 2H), 1.40-0.97 (m, 17H), 0.97-0.80 (m, 11H), 0.73-0.55 (m, 5H).

[1427] LCMS Rt=1.323 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.27H.sub.45 [M+H−2H.sub.2O].sup.+ 369, found 369.

Step 12

[1428] To a solution of A10-B75 (285 mg, 0.47 mmol) in THF (5 mL) and MeOH (5 mL) was added NaOH (200 mg, 5.00 mmol) and H.sub.2O (2 mL) at 25° C. Then the solution was stirred at 50° C. for 16 h. The reaction solution was extracted with EtOAc (2×10 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give crude product which was purified by a silica gel column (PE/EtOAc=3:1) to give a solid, which was then triturated in hot MeCN (2×1 mL) to give desired product Compound 1-B75 (18 mg, 10%) as a solid.

[1429] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.38-3.25 (m, 1H), 1.98-1.92 (m, 1H), 1.92-1.77 (m, 2H), 1.77-1.50 (m, 4H), 1.50-1.30 (m, 6H), 1.30-0.97 (m, 17H), 0.97-0.80 (m, 12H), 0.73-0.55 (m, 5H).

[1430] LCMS Rt=1.320 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.27H.sub.45 [M+H−2H.sub.2O].sup.+ 369, found 369.

Synthesis of 1A75 to Confirm Stereochemistry

[1431] ##STR00429##

[1432] To THF (0.5 mL) under N.sub.2 at −70° C. was added n-BuLi (2.5 M, 2.18 mmol, 0.872 mL). After that, a suspension of E-26781 (400 mg, 0.875 mmol) in THF (3 mL) was added drop-wise to give a suspension. After stirring at −70° C. for 30 min, a solution of (2R)-2-(propan-2-yl)oxirane (112 mg, 1.31 mmol) in THF (0.5 mL) was added. Then reaction was stirred at stirred at 25° C. for 16 hours. The mixture was poured into ice-water (20 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered and concentrated in vacuum to afford E-2678_2 (380 mg, crude) as a solid, which was used directly in the next step.

[1433] To a solution of E-2678_2 (380 mg, 0.7 mmol) and nickel (II) chloride (4.53 mg, 0.035 mmol) in MeOH (30 mL) was added Mg powder (336 mg, 14 mmol) was added at 25° C. The mixture was stirred at 50° C. for 1 h. After cooling, the mixture was quenched with HCl (100 mL, 2M) until the reaction became clear and extracted with EtOAc (2×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered concentrated and purified by a silica gel column (PE/EtOAc=10/1 to 3/1) to give E-2678_3 (140 mg, 50%) as a solid.

[1434] The E-2678_3 (140 mg, 0.347 mmol) was separated by SFC (column: AD (250 mm*30 mm, um)), gradient: 40-40% B (A=0.1% NH.sub.3H.sub.2O ETOH), flow rate: 50 mL/min) to give E-2678_3 (95 mg, 34%) as a solid.

[1435] .sup.1HNMR (400 MHz, CDCl.sub.3) δ.sub.H 5.42-5.38 (m, 1H), 3.35-3.26 (m, 1H), 2.20-2.14 (m, 1H), 2.10-1.89 (m, 4H), 1.87-1.59 (m, 6H), 1.54-1.16 (m, 12H), 1.13-0.97 (m, 7H), 0.95-0.75 (m, 11H), 0.68 (s, 3H).

[1436] SFC Rt=5.305 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 98.5% de. To a solution of E-26783 (95 mg, 0.235 mmol) in MeOH (10 mL) was added Pd/C (0.1 g, <1% water). Then the solution was hydrogenated under 50 psi of hydrogen at 50° C. for 16 hrs. The mixture was filtered through a pad of celite and the filtrate was concentrated in vacuum. The residue was purified by combi-flash (0-15% of EtOAc in PE) to afford DA ST-200-094-006 (23 mg, 24%) as a solid.

[1437] .sup.1HNMR (400 MHz, CDCl.sub.3) δ.sub.H 3.35-3.28 (m, 1H), 1.99-1.62 (m, 7H), 1.55-1.33 (m, 7H), 1.31-1.20 (m, 7H), 1.17-0.97 (m, 10H), 0.93-0.75 (m, 11H), 0.73-0.65 (m, 5H).

[1438] LCMS Rt=1.269 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.27H.sub.45 [M−2H.sub.2O+H]=369, found 369.

Example 76. Syntheses of Compounds 276, 376, and 476

[1439] ##STR00430## ##STR00431##

Step 1

[1440] To a solution of BHT (41.9 g, 190.58 mmol) in toluene (100 mL) under N.sub.2 at 0° C. was added AlMe.sub.3 (47.6 mL, 2 M in toluene, 95.2 mmol) drop-wise. The mixture was stirred at 25° C. for 1 h. To the mixture was added a solution of A376 (9.21 g, 33.6 mmol) in DCM (30 mL) at −78° C. After stirring at −78° C. for 1 h, EtMgBr (33.3 mL, 100 mmol) was added at −78° C. The mixture was stirred at −78° C. for 1.5 h. The reaction mixture was treated with saturated citric acid (50 mL). The organic phase was separated, extracted with EtOAc (80 mL). The organic phase was washed with brine (2×100 mL), dried over Na.sub.2SO.sub.4, concentrated in vacuum to give A1176 (5.2 g, 51%) as a solid.

[1441] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.47-2.40 (m, 1H), 2.11-2.00 (m, 1H), 1.96-1.77 (m, 6H), 1.69-1.63 (m, 2H), 1.58-1.50 (m, 3H), 1.38-1.20 (m, 6H), 1.56-0.97 (m, 5H), 0.90-0.87 (m, 6H), 0.80-0.64 (m, 2H).

Step 2

[1442] To a suspension of PPh.sub.3EtBr (18.2 g, 49.1 mmol) in THF (20 mL) was added t-BuOK (5.50 g, 49.1 mmol) at 20° C. After stirring at 40° C. for 30 mins, a solution of A1176 (5 g, 16.4 mmol) in THF (20 mL) was added at 40° C. and the reaction mixture was stirred at 40° C. for 1 h. The reaction mixture was poured into 50 g of crushed ice and stirred for 15 minutes. The organic layer was separated and the water phase was extracted with EtOAc (30 mL). The combined organic phase was concentrated in vacuum to give thick oil. The residue was dissolved in 90 mL of MeOH at 60° C., following by treating with 90 mL of water and a large amount of a precipitate appeared. After stirring at 60° C. for 1 h, the precipitate was collected by filtration and washed with a solution of MeOH/water (15 mL/15 mL), dried in vacuum to give A1276 (5.0 g, 96%) as a solid.

[1443] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.14-5.09 (m, 1H), 2.39-2.33 (m, 1H), 2.25-2.14 (m, 2H), 1.87-1.75 (m, 3H), 1.66-1.63 (m, 5H), 1.58-1.54 (m, 5H), 1.34-1.25 (m, 2H), 1.19-1.03 (m, 7H), 0.90-0.84 (m, 8H), 0.73-0.64 (m, 2H).

Step 3

[1444] To a solution of A1276 (4.5 g, 14.2 mmol) and methyl propiolate (3.78 mL, 42.6 mmol) in anhydrous dichloromethane (100 mL) under N.sub.2 at 0° C. was added dropwise Et.sub.2AlCl (1.0 M in toluene, 56.8 mL, 56.8 mmol). The mixture was stirred at 20° C. for 16 hrs. The reaction mixture was quenched with aqueous saturated citric acid (100 mL) at 0° C. carefully, and the inner temperature was maintained below 10° C. The resultant mixture was filtered through a pad of celite and washed with DCM (2×200 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated to give impure A1376 as a solid. The crude product was combined with another batch of impure product from 0.5 g of A1276, purified by flash column (0-30% of EtOAc in PE) to give A1376 (6 g, 95%) as a solid.

[1445] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.93 (dd, J=8.0, 15.6 Hz, 1H), 5.80 (dd, J=1.2, 15.2 Hz, 1H), 5.39-5.39 (m, 1H), 3.73 (s, 3H), 3.02-2.97 (m, 1H), 2.07-2.01 (m, 1H), 1.85-1.52 (m, 8H), 1.35-0.96 (m, 10H), 0.90-0.84 (m, 8H), 0.76-0.66 (m, 6H).

Step 4

[1446] To a solution of A1376 (6 g, 14.9 mmol) in EtOAc (100 mL) was added Pd/C (1 g, 10%, wet) and the mixture was degassed with H.sub.2 for 3 times. After that, the reaction was stirred at 15° C. under 15 psi of H.sub.2 for 16 hrs. The reaction mixture was filtered through a pad of celite and washed with EtOAc (30 mL). The filtrate was concentrated to give Compound 276 (5.8 g, 96%) as a solid.

[1447] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.66 (s, 3H), 2.39-2.31 (m, 1H), 2.25-2.17 (m, 1H), 1.95-1.92 (m, 1H), 1.85-1.78 (m, 4H), 1.68-1.61 (m, 3H), 1.57-1.52 (m, 5H), 1.42-1.40 (m, 2H), 1.35-1.26 (m, 3H), 1.15-1.02 (m, 8H), 0.92-0.82 (m, 8H), 0.81-0.79 (m, 1H), 0.72-0.59 (m, 4H). LCMS R.sub.t=1.334 min in 2 min chromatography, 30-90AB, MS ESI calcd. For C.sub.26H.sub.43O.sub.2 [M+H−H.sub.2O].sup.+ 387, found 387.

Step 5

[1448] To a solution of Compound 276 (also 175 in Example 75) (200 mg, 0.494 mmol) in THF (40 mL) was added LAH (56.1 mg, 1.48 mmol) at 0° C. under N.sub.2. After stirring at this temperature for 1 h, the reaction mixture was treated with water (2 mL), adjusted to pH=1-2 with saturated citric acid. The water phase was extracted with EtOAc (2×50 mL). The combined organic layers were washed with brine (2×30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to give Compound 376 (120 mg, 65%) as a solid.

[1449] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.63-3.60 (m, 2H), 1.96-1.93 (m, 1H), 1.85-1.78 (m, 3H), 1.68-1.52 (m, 10H), 1.48-1.38 (m, 3H), 1.34-1.25 (m, 4H), 1.13-1.07 (m, 9H), 0.93-0.83 (m, 8H), 0.73-0.59 (m, 4H).

[1450] LCMS Rt=3.826 min in 2 min chromatography, 30-90AB, MS ESI calcd. For C.sub.25H.sub.43O [M+H−H.sub.2O].sup.+ 359, found 359.

Step 6

[1451] To a suspension of DMP (2.24 g, 5.30 mmol) in DCM (18 mL) was added a solution of Compound 376 (1 g, 2.65 mmol) in DCM (10 mL) at 20° C. The reaction was stirred for 1 h at 20° C. The mixture was quenched with saturated NaHCO.sub.3 aqueous (20 mL) at 20° C. The mixture was filtered and the organic layers were separated and the aqueous was extracted with DCM (2×20 mL). The combined phase was washed with saturated Na.sub.2S.sub.2O.sub.3 aqueous (50 mL), brine (40 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give the crude product, which was purified by flash column (0-30% of EtOAc in PE) to give A1476 (920 mg, 93%) as a solid.

[1452] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 9.76 (s, 1H), 2.47-2.41 (m, 1H), 2.40-2.32 (m, 1H), 1.97-1.92 (m, 1H), 1.86-1.78 (m, 3H), 1.68-1.52 (m, 6H), 1.44-1.41 (m, 1H), 1.35-1.22 (m, 6H), 1.12-1.03 (m, 8H), 0.92-0.79 (m, 10H), 0.78-0.57 (m, 4H).

Step 7

[1453] To a solution of A1476 (910 mg, 2.42 mmol) in THF (60 mL) was added iPrMgCl (12.1 mL, 24.2 mmol) at 0° C. The reaction was stirred at this temperature for 1 h. The reaction was quenched by adding water (30 mL) and saturated citric acid solution (30 mL). The mixture was extracted with EtOAc (2×30 mL). The combined organic layer was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrate to give an oil, which was purified by silica gel column (PE:EtOAc=50:1 to 4:1) to give Compound 476 (900 mg, 89%) as a solid.

[1454] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.32-3.30 (m, 1H), 1.96-1.93 (m, 1H), 1.85-1.78 (m, 3H), 1.68-1.52 (m, 12H), 1.43-1.02 (m, 16H), 0.93-0.83 (m, 13H), 0.70-0.61 (m, 4H). LCMS R.sub.t=1.371 min in 2 min chromatography, 30-90AB, MS ESI calcd. For C.sub.28H.sub.47 [M+H−2H.sub.2O].sup.+ 383, found 383.

Example 77. Syntheses of Compounds 4A77 and 4B77

[1455] ##STR00432## ##STR00433##

Step 1

[1456] To a solution of Compound 476 (800 mg, 1.91 mmol) in pyridine (20 mL) was added BzCl (402 mg, 2.86 mmol) at 0° C. and the reaction was stirred at 20° C. for 18 h. The reaction mixture was diluted with water (50 mL), extracted with EtOAc (2×40 mL). The organic layer was washed with brine (5×50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The crude was purified by silica gel column (PE/EtOAc=50/1 to 4/1) to give A15 (600 mg, 60%) as an oil.

[1457] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.05 (d, J=8.0 Hz, 2H), 7.55 (t, J=7.2 Hz, 1H), 7.45 (d, J=8.0 Hz, 2H), 5.00-4.92 (m, 1H), 2.05-1.20 (m, 13H), 1.20-0.75 (m, 30H), 0.75-0.50 (m, 5H). A15 was purified by SFC (column: AD (250 mm*30 mm, 5 um)), gradient: 40-40% B (A=NH3/H2O, B=MeOH), flow rate: 60 mL/min) to give A16-A (116 mg, 19.4%) and impure A16-B (230 mg).

[1458] A16-A:

[1459] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.05 (d, J=8.0 Hz, 2H), 7.55 (t, J=7.2 Hz, 1H), 7.45 (d, J=8.0 Hz, 2H), 5.00-4.92 (m, 1H), 2.05-1.83 (m, 2H), 1.83-1.20 (m, 11H) 1.20-0.75 (m, 3OH), 0.75-0.50 (m, 5H).

[1460] A16-B:

[1461] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.04 (d, J=8 Hz, 2H), 7.55 (t, J=7.2 Hz, 1H), 7.45 (d, J=8.0 Hz, 2H), 5.00-4.92 (m, 1H), 2.05-1.42 (m, 15H), 1.40-1.15 (m, 4H) 1.14-0.75 (m, 24H), 0.73-0.50 (m, 5H).

Step 2

[1462] To a solution of A16-A (116 mg, 221 μmol) in THF (2 mL) and MeOH (2 mL) was added LiOH (52.6 mg, 2.20 mmol) and H.sub.2O (1 mL) at 25° C. Then the solution was stirred at 50° C. for 24 h. The reaction solution was extracted with EtOAc (2×10 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give crude product (91 mg), which was purified by flash column (0˜5% of acetone in DCM, 25° C.) to give Compound 4A77 (45 mg, 50%) as a solid.

[1463] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.33-3.31 (m, 1H), 1.96-1.85 (m, 1H), 1.84-1.77 (m, 3H), 1.68-1.62 (m, 5H), 1.56-1.52 (m, 5H), 1.44-1.33 (m, 4H), 1.31-1.17 (m, 5H), 1.14-0.99 (m, 8H), 0.92-0.79 (m, 13H), 0.73-0.56 (m, 5H). LCMS Rt=1.347 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.28H.sub.47 [M+H−2H.sub.2O].sup.+ 383, found 383.

Step 3

[1464] To a solution of A16-B (130 mg, 248 μmol) in THF (2 mL) and MeOH (2 mL) and H.sub.2O (1 mL) was added lithium hydroxide hydrate (104 mg, 2.48 mmol) at 25° C. Then the solution was stirred at 50° C. for 16 h. The reaction was dilute with water (10 mL) and extracted with EtOAc (2×30 mL). The combined organic layers was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue was purified by silica gel chromatograph (PE/EtOAc=10/1) to afford Compound 4B77 (59 mg, 57%) as a solid.

[1465] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.33-3.31 (m, 1H), 1.97-1.88 (m, 1H), 1.84-1.77 (m, 3H), 1.68-1.59 (m, 6H), 1.55-1.52 (m, 5H), 1.46-1.37 (m, 1H), 1.33-1.15 (m, 6H), 1.14-0.99 (m, 9H), 0.94-0.79 (m, 12H), 0.85-0.78 (m, 1H), 0.73-0.56 (m, 5H). LCMS Rt=1.357 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.28H.sub.46 [M+H−2H.sub.2O].sup.+ 383, found 383.

Synthesis to Confirm Stereochemistry

[1466] ##STR00434##

[1467] To THF (0.5 mL) was added n-BuLi (2.5 M, 2.12 mmol, 0.848 mL) under N.sub.2 at −70° C. After that, a suspension of E-2863_1 (400 mg, 0.849 mmol) in THF (3 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, a solution of (2R)-2-(propan-2-yl)oxirane (86.9 mg, 1.01 mmol) in THF (0.5 mL) was added. The reaction was stirred at stirred at 25° C. for 16 hours. The mixture was poured into ice-water (20 mL) and extracted with EA (2×30 mL). The combined organic layers were washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to afford E-2863_2 (430 mg, crude) as a solid, which was used directly for the next step.

[1468] To a solution of E-2863_2 (430 mg, 0.772 mmol) and nickel (II) chloride (5 mg, 0.0386 mmol) in MeOH (30 mL) was added Mg powder (369 mg, 15.4 mmol) at 25° C. The mixture was stirred at 50° C. for 1 h. After cooling, the mixture was quenched with HCl (100 mL, 2M) until the reaction became clear and extracted with EtOAc (2×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by a silica gel column (PE/EtOAc=10/1 to 3/1) to give DA ST-200-094-002 (160 mg, 50%) as a solid, which was separated by SFC (column: AD (250 mm*30 mm, 5 um)), gradient: 40-40% B (A=0.1% NH.sub.3H.sub.2O ETOH), flow rate: 50 mL/min) to give DA ST-200-094-002 (85 mg, 53%, 50 mg for delivery) as a solid.

[1469] .sup.1HNMR (400 MHz, CDCl3) δ 5.42-5.38 (m, 1H), 3.35-3.26 (m, 1H), 2.25-2.21 (m, 1H), 2.07-1.77 (m, 7H), 1.70-1.59 (m, 3H), 1.54-1.36 (m, 7H), 1.32-0.99 (m, 11H), 0.96-0.75 (m, 14H), 0.68 (s, 3H).

[1470] LCMS Rt=1.291 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.28H.sub.47O [M−H.sub.2O+H]=399, found 399.

[1471] SFC Rt=5.654 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 96.8% de

[1472] To a solution of DA ST-200-094-002 (35 mg, 0.0839 mmol) in MeOH (6 mL) was added Pd/C (0.1 g, <1% water). Then the suspension was hydrogenated under 50 psi of hydrogen at 50° C. for 16 hrs. The mixture was filtered through a pad of celite and the filtrate was concentrated in vacuum. The residue was purified by flash column (PE/EtOAc=10/l to 5/1) to give DA ST-200-094-004 (7 mg, 20%) as a solid.

[1473] .sup.1HNMR (400 MHz, CDCl3) δ 3.35-3.28 (m, 1H), 2.00-1.91 (m, 1H), 1.87-1.74 (m, 3H), 1.71-1.56 (m, 6H), 1.54-1.35 (m, 8H), 1.32-1.17 (m, 5H), 1.14-0.94 (m, 9H), 0.93-0.76 (m, 13H), 0.73-0.62 (m, 4H).

[1474] LCMS Rt=1.350 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.28H.sub.47 [M−2H2O+H]=383, found 383.

Example 78. Synthesis of Compound 561

[1475] ##STR00435##

Step 1

[1476] t-BuOH (300 mL) was charged into a three-neck round bottom flask under nitrogen at 35° C. and stirred under nitrogen gas bubbling for 10 mins. t-BuOK (45.2 g, 403 mmol) was added to the mixture and stirred under nitrogen gas bubbling for 15 mins. A178 (10 g, 36.7 mmol) was added to the above mixture and stirred under nitrogen gas bubbling at 35° C. for 1.5 hrs. The reaction mixture was poured to 10% aqueous acetic acid (500 mL) and stirred for 15 mins. Water (200 mL) was added to the aqueous and stirred for 30 mins. The pH of the mixture was adjusted to 7˜8 with sodium bicarbonate (60 g). The mixture was stirred for 30 mins. The mixture was extracted with PE (3×400 mL). The organic layer was separated, washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered and concentrated below 40° C. to give A1878 (11 g, crude) as an oil.

[1477] .sup.1H NMR CDCl.sub.3 (400 MHz, CDCl.sub.3) δ 5.55-5.47 (m, 1H), 3.16-2.94 (m, 2H), 2.52-2.33 (m, 4H), 2.19-1.93 (m, 6H), 1.75-1.61 (m, 2H), 1.56-1.48 (m, 1H), 1.40-1.33 (m, 3H), 1.29-1.22 (m, 1H), 1.01-0.92 (m, 4H).

Step 2

[1478] To solution of BHT (52.3 g, 238 mmol) in anhydrous toluene (150 mL) under N.sub.2 at 0° C. was added trimethylaluminum (2 M in toluene, 55.0 mL, 110 mmol) drop-wise. The mixture was stirred at 15° C. for 1 hour and cooled to −70° C. Then A1878 (10 g, 36.7 mmol) in toluene (50 mL) was added below −60° C. The resulting mixture was stirred at −70° C. for 1 hour. Ethylmagnesium bromide (36.6 mL, 3.0 M in diethyl ether, 110 mmol) was added drop-wise below −60° C. The reaction mixture was stirred at −70° C. for another 1 hour. The reaction mixture was quenched with saturated citric acid (400 mL) at −70° C. The mixture was warmed to 15° C. slowly and extracted with ethyl acetate (3×400 mL). The combined organic layer was washed with brine (500 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by Combi-flash (0%˜30% of EtOAc in PE) to afford A1978 (7.6 g, 69%) as a solid.

[1479] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.39-5.32 (m, 1H), 2.43-2.33 (m, 1H), 2.22-2.15 (m, 1H), 2.05-1.86 (m, 6H), 1.80-1.70 (m, 2H), 1.65-1.52 (m, 2H), 1.47-1.29 (m, 5H), 1.26-1.13 (m, 4H), 0.85-0.76 (m, 8H).

Step 3

[1480] To a suspension of PPh.sub.3EtBr (38.9 g, 105 mmol) in THF (200 mL) under N.sub.2 was added t-BuOK (11.7 g, 105 mmol) at 40° C. After stirring at 20° C. for 10 min, A19 (8 g, 26.4 mmol) was added. The reaction mixture was stirred at 40° C. for 1 h. The reaction was quenched with aqueous NH.sub.4Cl (250 mL) at 0° C. and extracted with EtOAc (3×200 mL). The combined organic phase was washed with brine (500 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by Combi-flash (0%˜30% of EtOAc in PE) to afford A2078 (7.2 g, 87%) as a solid.

[1481] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.36-5.29 (m, 1H), 5.12-5.01 (m, 1H), 2.36-2.25 (m, 1H), 2.23-2.05 (m, 3H), 2.00-1.73 (m, 5H), 1.62-1.48 (m, 7H), 1.43-1.32 (m, 3H), 1.28-1.06 (m, 5H), 0.86-0.73 (m, 8H).

Step 4

[1482] To a solution of A2078 (7 g, 22.2 mmol) and methyl propiolate (4.66 g, 55.5 mmol) in DCM (200 mL) were added diethyl aluminum chloride (88.8 mL, 88.8 mmol, 1 M in hexane) at 0° C. under N.sub.2 drop-wise. The reaction mixture was stirred at 25° C. for 16 h. The reaction mixture was quenched with saturated aqueous NaHCO.sub.3 (100 mL) solution, acidified with saturated aqueous citric acid solution to pH=5, extracted with DCM (2×200 mL). The combined organic layer was washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give a crude product. The crude product was purified by silica gel column (PE/EtOAc=4/1) to give A2178 (6.20 g, 70%) as a solid.

[1483] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.00-6.90 (m, 1H), 5.85-5.75 (m, 1H), 5.40-5.30 (m, 2H), 3.73 (s, 3H), 3.05-2.95 (m, 1H), 2.30-2.20 (m, 1H), 2.10-1.75 (m, 9H), 1.75-1.50 (m, 3H), 1.50-1.20 (m, 9H), 0.95-0.80 (m, 5H), 0.78 (s, 3H).

Step 5

[1484] To a solution of A2178 (800 mg, 2.00 mmol) in EtOAc (50 mL) was added lindlar catalyst (500 mg) and the reaction mixture was stirred at 20° C. for 4 h under H.sub.2. The reaction mixture was filtered with filter paper and concentrated in vacuum to give a crude product. The crude product was purified by silica gel column (PE/EtOAc=10/1) to give A2278 (650 mg, crude).

[1485] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.35 (m, 2H), 3.66 (s, 3H), 2.50-2.40 (m, 1H), 2.35-2.25 (m, 2H), 2.15-2.05 (m, 1H), 2.05-1.95 (m, 3H), 1.95-1.75 (m, 3H), 1.75-1.55 (m, 3H), 1.55-1.40 (m, 7H), 1.40-1.25 (m, 3H), 1.10-1.00 (m, 4H), 1.00-0.85 (m, 4H), 0.85-0.80 (m, 1H), 0.75 (s, 3H).

Step 6

[1486] To a solution of A22 (300 mg, 0.748 mmol) in THF (10 mL) was added lindlar catalyst (500 mg) and the reaction mixture was stirred at 20° C. for 4 h under H.sub.2. The reaction mixture was filtered with filter paper and concentrated in vacuum to give a crude product. The crude product was purified by silica gel column (PE/EtOAc=10/1) to give an impure product. The impure product was purified by prep-HPLC (0.1% TFA as additive). Most of MeCN was removed by concentration and the remaining solvent was removed by lyophilization to give 561 (27 mg, 9%) as a solid.

[1487] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 3.66 (s, 3H), 2.40-2.30 (m, 1H), 2.30-2.20 (m, 2H), 2.10-1.80 (m, 8H), 1.55-1.40 (m, 6H), 1.40-1.20 (m, 5H), 1.20-1.00 (m, 5H), 1.00-0.90 (m, 3H), 0.90-0.75 (m, 4H), 0.75-0.70 (m, 1H), 0.68 (s, 3H). LCMS Rt=1.299 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.26H.sub.41O.sub.2[M+H−H.sub.2O].sup.+ 385, found 385.

Example 79. Synthesis of Compound 679

[1488] ##STR00436##

[1489] To a solution of 276 (150 mg, 0.37 mmol) in THF (5 mL) was added MeMgBr (616 μL, 3 M in ether) drop-wise at 0° C. under N.sub.2. After that, the reaction mixture was stirred at 20° C. for 1 h. The reaction mixture was quenched with saturated aqueous NH.sub.4Cl (15 mL) solution, extracted with EtOAc (2×20 mL). The combined organic layer was washed with brine (20 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give a crude product. The crude product was re-crystallized from MeCN (10 mL) to give 679 (32 mg, 21%) as a solid.

[1490] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.05-1.95 (m, 1H), 1.90-1.80 (m, 3H), 1.65-1.60 (m, 3H), 1.60-1.55 (m, 2H), 1.45-1.25 (m, 8H), 1.25-1.15 (m, 8H), 1.15-1.00 (m, 10H), 0.95-0.80 (m, 8H), 0.75-0.55 (m, 5H). LCMS Rt=1.282 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.27H.sub.45 [M+H−2H.sub.2O].sup.+ 369, found 369.

Example 80. Synthesis of Compound 780

[1491] ##STR00437##

[1492] To a solution of 561 (300 mg, impure, 0.745 mmol) in THF (10 mL) was added MeLi (2.32 mL, 3.72 mmol, 1.6M in THF). The mixture was stirred at 25° C. for 30 minutes. The mixture was quenched with sat. NH.sub.4Cl (30 mL) and extracted with EtOAc (3×15 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-15% of EtOAc in PE) to give 780 (37 mg, 12%) as a solid.

[1493] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 2.26-2.20 (m, 1H), 2.10-1.75 (m, 7H), 1.68-1.58 (m, 2H), 1.56-1.37 (m, 7H), 1.36-1.24 (m, 4H), 1.23-1.17 (m, 8H), 1.16-0.99 (m, 5H), 0.96-0.90 (m, 3H), 0.89-0.76 (m, 5H), 0.68 (s, 3H).

[1494] LCMS Rt=1.222 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.27H.sub.43 [M+H−2H.sub.2O].sup.+ 367, found 367.

Example 81: Synthesis of 8127

[1495] ##STR00438##

[1496] The experimental of intermediate 200-N19-2_4 or 276 can be found in Example 76.

Synthesis of 8127

[1497] ##STR00439##

[1498] Ti(i-PrO).sub.4 (140 mg, 0.5 mmol) and EtMgBr (0.6 mL, 3 M in Et.sub.2O, 1.72 mmol) were added to a solution of 200-N19-2_4 (200 mg, 0.5 mmol) in THF (2 mL) at 25° C. After that, the reaction mixture was stirred at 25° C. for 15 min under N.sub.2. The reaction mixture was quenched with saturated aqueous NH.sub.4Cl (10 mL) solution and extracted with EtOAc (3×20 mL). The combined organic layer was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give a crude product. The crude product was purified by silica gel column (EtOAc/PE=5/1) to afford an impure product, which was triturated from n-hexane (5 mL) at 25° C. to give 8127 (58 mg, 46%).

[1499] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.99-1.91 (m, 1H), 1.88-1.59 (m, 10H), 1.48-1.21 (m, 6H), 1.18-0.97 (m, 10H), 0.94-0.81 (m, 9H), 0.79-0.56 (m, 8H), 0.47-0.38 (m, 2H).

[1500] LCMS Rt=1.356 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. For C.sub.27H.sub.43 [M+H−2H.sub.2O].sup.+ 367, found 367.

Example 82: Synthesis of 8245

[1501] ##STR00440##

Synthesis of ST-200-6-16_1

[1502] ##STR00441##

[1503] The synthesis for ST-200-N19-3_5A can be found in Example 78. Ti(i-PrO).sub.4 (212 mg, 0.75 mmol) was added to a solution of 200-N19-3_5A (300 mg, 0.75 mmol) in THF (2.5 mL), followed by adding EtMgBr (0.9 mL, 3 M in Et.sub.2O, 2.6 mmol) at 25° C. Next, the reaction mixture was stirred at 25° C. for 15 min under N.sub.2. The reaction mixture was quenched with saturated aqueous NH.sub.4Cl (10 mL) solution and extracted with EtOAc (20 mL×3). The combined organic layer was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give a crude product. The crude product was purified by silica gel column (EtOAc/PE=5/1) to afford a crude product as a solid, which was purified by re-crystallized from MeCN (5 mL) at 85° C. to give impure product as a solid. The impure sample was further purification by SFC (column: OD (250 mm*30 mm, 10 um)), gradient: 25-25% B (0.1% NH.sub.3H.sub.2O ETOH), flow rate: 60 mL/min) to give ST-200-6-16_1 (110 mg, 44%).

[1504] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.44-5.38 (m, 1H), 5.33-5.28 (m, 1H), 2.29-2.21 (m, 1H), 2.15-1.94 (m, 5H), 1.93-1.79 (m, 3H), 1.78-1.57 (m, 6H), 1.52-1.21 (m, 10H), 1.06-0.98 (m, 3H), 0.92-0.82 (m, 5H), 0.77 (s, 3H), 0.75-0.69 (m, 2H), 0.47-0.39 (m, 2H).

[1505] LCMS Rt=1.219 min in 2.0 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.41O [M+H−H.sub.2O].sup.+ 381, found 381.

Synthesis of 8245

[1506] ##STR00442##

[1507] Lindlar catalyst (100 mg) was added to a solution of ST-200-6-16_1 (78 mg, 0.2 mmol) in THF (5 mL) and the mixture was degassed and backed-filled with H.sub.2 3 times. After that, the reaction mixture was stirred at 25° C. for 4 h under H.sub.2. The reaction mixture was filtered through a pad of celite washed with THF (100 mL) and concentrated in vacuum to give a crude product, which was re-crystallized from MeCN (5 mL) at 85° C. to give ST-200-6-16 (32 mg, 41%).

[1508] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.43-5.34 (m, 1H), 2.27-2.19 (m, 1H), 2.07-1.72 (m, 8H), 1.67-1.59 (m, 3H), 1.55-1.37 (m, 7H), 1.34-0.96 (m, 10H), 0.94-0.91 (m, 3H), 0.89-0.82 (m, 4H), 0.76-0.70 (m, 2H), 0.68 (s, 3H), 0.48-0.37 (m, 2H).

[1509] LCMS Rt=1.186 min in 2.0 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.43O [M+H−H.sub.2O].sup.+ 383, found 383.

Example 83: Synthesis of 8361, 8378, and 8379

[1510] ##STR00443## ##STR00444##

[1511] The synthesis of 200-N19-4_5 can be found in Example 94.

Synthesis of 200-N19-4_6

[1512] ##STR00445##

[1513] KI (28.0 g, 169 mmol) was added to a solution of 200-N19-4_5 (17 g, 33.9 mmol) in DMF (200 mL) at 25° C. The mixture was stirred at 50° C. for 2 hours. Half of the reaction mixture was poured into water (500 mL). The suspension was extracted with PE (700 mL). The combined organic phase was washed with saturated brine (2×500 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give 200-N19-4_6 (8.5 g, crude) as an oil. The other half of the reaction mixture was used directly for the next step.

[1514] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 3.35-3.30 (m, 1H), 3.20-3.10 (m, 1H), 2.25-2.15 (m, 1H), 2.05-1.76 (m, 8H), 1.69-1.34 (m, 9H), 1.30-1.13 (m, 7H). 0.92-0.75 (m, 6H). 0.71 (s, 3H).

Synthesis of 200-N19-4_7

[1515] ##STR00446##

[1516] PhSO.sub.2Na (9.15 g, 55.8 mmol) was added to the reaction mixture from the previous step at 25° C. The mixture was stirred at 50° C. for 3 hours. The reaction mixture was poured into water (500 ml) and some solid was produced. The mixture was filtered. The filter cake was washed with water (2×500 ml). The resulting filter cake was dissolved in DCM (500 mL), washed with water (2×500 mL). The combined organic phase was washed with saturated brine (2×500 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give 200-N19-4_7 (8.5 g, crude) as a solid, which was re-crystallized from MeCN (50 mL) at reflux (82° C.). After cooling to 25° C., the mixture was filtered and concentrated in vacuum to get 200-N19-4_7 (5 g, 59%) as a solid. Mother liquor filtered and concentrated to give another 2 g of solid.

[1517] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.92-7.88 (m, 2H), 7.65-7.53 (m, 3H), 5.38-5.33 (m, 1H), 3.18-3.10 (m, 1H), 2.90-2.80 (m, 1H), 2.25-2.16 (m, 1H), 1.88-1.60 (m, 9H), 1.59-1.35 (m, 5H), 1.29-1.05 (m, 11H), 0.88-0.77 (m, 5H), 0.65 (s, 3H).

Synthesis of 200-N19-6-14_1

[1518] ##STR00447##

[1519] n-BuLi (2 mL, 2.5 M, 5.08 mmol) was added to a solution of diisopropylamine (0.73 mL, 5.08 mmol) in THF (1 mL) under N.sub.2 at −70° C. The resulting mixture was warmed to 25° C. and stirred at 25° C. for 30 min. After re-cooling to −70° C., a solution of 200-N19-4_6 (0.6 g, 1.27 mmol) in THF (3 mL) was added at −70° C. The reaction mixture was stirred at −70° C. for 1 hour. 2-(tert-butyl)oxirane (152 mg, 1.52 mmol) was added at −70° C. The reaction mixture was warmed to 25° C. and stirred at 25° C. for 18 hours. The reaction mixture was quenched with saturated NH.sub.4Cl aqueous (10 mL) at 0° C., extracted with EtOAc (2×10 mL). The combined organic phase was washed with brine (2×10 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a crude product, which was purified by flash column (0-30% of PE in EtOAc, 50 mins) to give 200-N19-6-14_1 (550 mg) as a solid, which was used directly for the next step.

Synthesis of 200-N19-6-14_2 (8361)

[1520] ##STR00448##

[1521] Mg (1.16 g, powder) was added to a solution of 200-N19-6-14_1 (550 mg, crude) in MeOH (40 mL) at 65° C. The mixture was stirred at 65° C. for 3 hours and quenched by adding HCl (50 mL, 2 M in water). The mixture was extracted with EtOAc (2×50 mL). The organic layers were washed with water (2×100 mL), sat. NaHCO.sub.3 (2×100 mL), brine (2×80 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to give a crude product, which was purified by flash column (0-20% of PE in EtOAc, 60 mins) to give 200-N19-6-14_2 (190 mg) as a solid.

[1522] 200-N19-6-14_2 (45 mg) was re-crystallized from MeCN at 70° C. to give 200-N19-6-14_2 (35 mg) as a solid.

[1523] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.30 (m, 1H), 3.40-3.00 (m, 1H), 2.30-2.15 (m, 1H), 2.10-1.60 (m, 9H), 1.55-1.40 (m, 6H), 1.25-1.00 (m, 11H), 0.95-0.75 (m, 18H), 0.68 (s, 3H).

[1524] LCMS Rt=1.338 min in 2 min chromatography, 30-90AB_E, purity 98%, MS ESI calcd. For C.sub.29H.sub.49O [M+H−H.sub.2O].sup.+ 413.

Synthesis of 8378 and 8379

[1525] ##STR00449##

[1526] 200-N19-6-14_2 (145 mg) was separated by SFC (Column: AD (250 mm*30 mm, 10 um); Condition: 0.1% NH.sub.3H.sub.2O ETOH, 40% B; FlowRate(ml/min): 60) to give impure DA-6-14 (70 mg) and impure DA-6-15 (60 mg) both as a solid. The impure DA-6-15 (60 mg) was triturated with MeCN (5 mL) at 25° C. to give DA-6-15 (27 mg, pure) as a solid. The impure DA-6-14 (70 mg) was triturated with MeCN (5 mL) at 25° C. to give DA-6-14 (27 mg, pure) as a solid.

[1527] 8378:

[1528] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.30 (m, 1H), 3.40-3.00 (m, 1H), 2.30-2.15 (m, 1H), 2.10-1.60 (m, 10H), 1.55-1.40 (m, 5H), 1.25-1.00 (m, 11H), 0.95-0.75 (m, 18H), 0.68 (s, 3H).

[1529] LCMS Rt=1.334 min in 2 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. For C.sub.29H.sub.47 [M+H−2H.sub.2O].sup.+ 395.

[1530] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.30 (m, 1H), 3.40-3.25 (m, 1H), 2.25-2.15 (m, 1H), 2.05-1.75 (m, 7H), 1.70-1.60 (m, 2H), 1.50-1.30 (m, 8H), 1.20-1.05 (m, 11H), 1.00-0.75 (m, 16H), 0.68 (s, 3H).

[1531] LCMS Rt=1.327 min in 2 min chromatography, 30-90AB_E, purity 99%, MS ESI calcd. For C.sub.28H.sub.47 [M+H−2H.sub.2O].sup.+ 395.

Synthesis of 8378 to Determine Stereochemistry

[1532] ##STR00450##

[1533] To a solution of diisopropylamine (0.262 mL, 1.82 mmol) in THF (0.5 mL) under N.sub.2 at −78° C. was added n-BuLi (0.676 mL, 2.5 M, 1.69 mmol). The resulting mixture was warmed to 0° C. and stirred at 0° C. for 10 mins. After re-cooling to −78° C., a solution of E-28781 (200 mg, 0.425 mmol) in THF (1.5 mL) was added at −78° C. The reaction mixture was stirred at −78° C. for 1 hr. (R)-2-(tert-butyl)oxirane (51.0 mg, 0.510 mmol) was added at −78° C. The reaction mixture was warmed to 25° C. and stirred at 25° C. for 16 hrs. The reaction mixture was quenched with saturated NH.sub.4Cl aqueous (10 mL) at 0° C., extracted with EtOAc (2×10 mL). The combined organic phase was washed with brine (2×10 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give E-2878_2 (250 mg, crude) as a solid, which was used directly for the next step.

[1534] To a solution of E-2878_2 (250 mg, crude) in MeOH (30 mL) was added Mg powder (840 mg, 35.0 mmol) and NiCl.sub.2 (20 mg) at 60° C. The mixture was stirred at 60° C. for 5 hrs. The reaction was quenched by HCl (50 mL, 2 M in water). The mixture was extracted with EtOAc (2×50 mL). The organic layers were washed with water (2×100 mL), sat. NaHCO.sub.3 (2×100 mL), brine (2×80 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude product which was purified by flash column (0-20% of EtOAc in PE) to give ST-200-094-007 (100 mg, 53%, impure, 96% de) as a solid. The impure product was re-purified by SFC separation (column: AD (250 mm*30 mm, 5 um), condition: 0.1% NH.sub.3H.sub.2O EtOH, Begin B: 40%, End B: 40%) to give ST-200-094-007 (70 mg, 100% de, impure) as a solid. The ST-200-094-007 (70 mg) was purified by recrystallization (n-BuOH/H.sub.2O=4/l) to give ST-200-094-007 (9 mg, pure,) as a solid and ST-200-094-007 (60 mg, impure) as a solid.

[1535] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.30 (m, 1H), 3.40-3.00 (m, 1H), 2.30-2.15 (m, 1H), 2.10-1.60 (m, 10H), 1.55-1.40 (m, 5H), 1.25-1.00 (m, 11H), 0.95-0.92 (m, 4H), 0.90 (s, 9H), 0.88-0.82 (m, 5H), 0.68 (s, 3H).

[1536] LCMS Rt=1.334 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.29H.sub.47 [M+H−2H.sub.2O].sup.+ 395, found 395.

[1537] SFC Rt=5.182 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100% de.

Example 84: Synthesis of 8462 and 8463

[1538] ##STR00451##

[1539] The synthesis of 200-N19-4_6 can be found in Example 83.

[1540] During the synthesis of 8462 from chiral epoxide, 8462 was proved to be S-configuration at C24 and 8463 was proved to be R-configuration at C24. See below.

Synthesis of 200-N19-4_6_1

[1541] ##STR00452##

[1542] To THF (5 mL) under N.sub.2 at −70° C. was added n-BuLi (4.23 mL, 2.5 M in n-hexane, 10.6 mmol). To the mixture was added dropwise a solution of ST-200-N19-4_6 (2 g, 4.24 mmol) in THF (15 mL) at −70° C. The reaction mixture was stirred at −70° C. for 1 hour. 2-isopropyloxirane (437 mg, 5.08 mmol) was added at −70° C. The reaction mixture was warmed to 25° C. slowly and stirred at 25° C. for 16 hours. The reaction mixture was quenched with saturated NH.sub.4Cl aqueous (50 mL). The mixture was extracted with EtOAc (2×30 mL). The combined organic phase was washed with brine (2×30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give st-200-N19-4_6_1 (2 g, crude) as an oil, which was used for the next step directly.

Synthesis of DA-6-6

[1543] ##STR00453##

[1544] To a solution of 200-N19-4_6_1 (2 g, 3.59 mmol) in MeOH (100 mL) was added Mg (4.35 g, 179 mmol) at 65° C. The mixture was stirred at 65° C. for 1 h. Aq. HCl (70 mL, 2 M in water) was added. The mixture was extracted with EtOAc (3×50 mL), washed with sat. NaHCO.sub.3 (2×150 mL), brine (2×100 mL) to give a crude product, which was purified by flash column (0-10% of EtOAc in PE, 60 mins) to give DA-6-6 (600 mg, 40%) as a solid.

Synthesis of 8462 and 8463

[1545] ##STR00454##

[1546] DA-6-6 (600 mg) was separated by SFC (Column: AD (250 mm*30 mm, 10 um); Condition: 0.1% NH.sub.3H.sub.2O ETOH, 40% B; FlowRate(ml/min): 60) to give 8462 (152 mg, 25%) and 8463 (137 mg, 23%) as a solid.

[1547] 8462

[1548] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.35 (m, 1H), 3.37-3.25 (m, 1H), 2.27-2.19 (m, 1H), 2.08-1.87 (m, 4H), 1.86-1.75 (m, 3H), 1.71-1.58 (m, 3H), 1.52-1.35 (m, 6H), 1.34-1.17 (m, 7H), 1.16-0.97 (m, 5H), 0.97-0.90 (m, 8H), 0.89-0.82 (m, 5H), 0.81-0.75 (m, 1H), 0.68 (s, 3H).

[1549] LCMS Rt=1.285 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. for C.sub.28H.sub.47O [M+H−H.sub.2O].sup.+ 399, found 399.

[1550] 8463

[1551] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.41-5.35 (m, 1H), 3.37-3.25 (m, 1H), 2.27-2.19 (m, 1H), 2.07-1.93 (m, 3H), 1.90-1.78 (m, 3H), 1.63-1.50 (m, 5H), 1.48-1.34 (m, 7H), 1.33-1.15 (m, 7H), 1.14-0.98 (m, 3H), 0.98-0.89 (m, 9H), 0.88-0.82 (m, 4H), 0.81-0.75 (m, 1H), 0.68 (s, 3H).

[1552] LCMS Rt=1.278 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. for C.sub.28H.sub.47O [M+H−H.sub.2O].sup.+ 399, found 399.

Synthesis to Confirm Stereochemistry

[1553] ##STR00455##

[1554] To THF (0.5 mL) was added n-BuLi (2.5 M, 2.12 mmol, 0.848 mL) under N.sub.2 at −70° C. After that, a suspension of E-2863_1 (400 mg, 0.849 mmol) in THF (3 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, a solution of (2R)-2-(propan-2-yl)oxirane (86.9 mg, 1.01 mmol) in THF (0.5 mL) was added. The reaction was stirred at stirred at 25° C. for 16 hours. The mixture was poured into ice-water (20 mL) and extracted with EA (2×30 mL). The combined organic layers were washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to afford E-2863_2 (430 mg, crude) as a solid, which was used directly for the next step.

[1555] To a solution of E-2863_2 (430 mg, 0.772 mmol) and nickel (II) chloride (5 mg, 0.0386 mmol) in MeOH (30 mL) was added Mg powder (369 mg, 15.4 mmol) at 25° C. The mixture was stirred at 50° C. for 1 h. After cooling, the mixture was quenched with HCl (100 mL, 2M) until the reaction became clear and extracted with EtOAc (2×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by a silica gel column (PE/EtOAc=10/1 to 3/1) to give DA ST-200-094-002 (160 mg, 50%) as a solid, which was separated by SFC (column: AD (250 mm*30 mm, 5 um)), gradient: 40-40% B (A=0.1% NH.sub.3H.sub.2O ETOH), flow rate: 50 mL/min) to give DA ST-200-094-002 (85 mg, 53%, 50 mg for delivery) as a solid.

[1556] .sup.1HNMR (400 MHz, CDCl3) δ 5.42-5.38 (m, 1H), 3.35-3.26 (m, 1H), 2.25-2.21 (m, 1H), 2.07-1.77 (m, 7H), 1.70-1.59 (m, 3H), 1.54-1.36 (m, 7H), 1.32-0.99 (m, 11H), 0.96-0.75 (m, 14H), 0.68 (s, 3H).

[1557] LCMS Rt=1.291 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.28H.sub.47O [M−H.sub.2O+H]=399, found 399.

Example 85: Synthesis of 8564, 8584, and 8585

[1558] ##STR00456##

[1559] The experimental of intermediate ST-200-N19-4_6 can be found in Example 83.

Synthesis of ST-200-N19-L5_1

[1560] ##STR00457##

[1561] A suspension of ST-200-N19-4_6 (500 mg, 1.06 mmol) in THF (4 mL) was added dropwise to a solution of n-BuLi (1.05 mL, 2.5 M in hexane, 2.65 mmol) in THF (1 mL) at −70° C. under N.sub.2. After stirring for 30 minutes at −70° C., a solution of diisopropylamine (267 mg, 2.65 mmol) was added dropwise at −70° C., followed by adding a solution of 2-ethyloxirane (114 mg, 1.59 mmol) dropwise at −70° C. The mixture was stirred at −70° C. for another 30 min and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 24 hours, quenched by saturated NH.sub.4Cl aqueous (5 mL), extracted with EtOAc (3×10 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-N19-L5_1 (610 mg, crude) as a solid, which was used directly.

Synthesis of ST-200-N19-L5

[1562] ##STR00458##

[1563] Mg powder (1.07 g, 44.8 mmol) and NiCl.sub.2 (5 mg, 0.05 mmol) were added to a solution of ST-200-N19-L5_1 (610 mg, 1.1 mmol) in 100 mL anhydrous MeOH and stirred under N.sub.2 at 60° C. The reaction mixture was quenched by 2 M HCl (50 mL) until solid was dissolved. The mixture was extracted with EtOAc (3×100 mL). The combined organic layer was washed with sat. NaHCO.sub.3 (150 mL), brine (150 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-15% of EtOAc in PE) to give ST-200-N19-L5 (240 mg, 53%) as a solid.

[1564] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.34 (m, 1H), 3.54-3.42 (m, 1H), 2.28-2.18 (m, 1H), 2.08-1.61 (m, 8H), 1.55-1.36 (m, 10H), 1.34-0.98 (m, 11H), 0.97-0.73 (m, 11H), 0.71-0.63 (m, 3H).

[1565] LCMS Rt=1.349 min in 2.0 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.45O [M+H−H.sub.2O].sup.+ 385, found 385.

Synthesis of ST-200-N19-L5R & ST-200-N19-L5S

[1566] ##STR00459##

[1567] ST-200-N19-L5 (208 mg, 0.52 mmol) was purified by SFC (column: AD (250 mm*30 mm, 10 um)), gradient: 40-40% B (A=0.1% NH.sub.3/H.sub.2O, B=EtOH), flow rate: 60 mL/min) to give ST-200-N19-L5R (80 mg, 38%) and ST-200-N19-L5S (70 mg, 33%) as a solid.

[1568] 8584

[1569] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.36 (m, 1H), 3.52-3.41 (m, 1H), 2.27-2.19 (m, 1H), 2.07-1.77 (m, 7H), 1.67-1.56 (m, 2H), 1.54-1.38 (m, 8H), 1.33-0.98 (m, 12H), 0.97-0.90 (m, 6H), 0.88-0.79 (m, 5H), 0.68 (s, 3H).

[1570] LCMS Rt=1.353 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.45O [M+H−H.sub.2O].sup.+ 385, found 385. SFC Rt=5.762 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, purity: 100%.

[1571] 8585

[1572] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.36 (m, 1H), 3.53-3.43 (m, 1H), 2.27-2.19 (m, 1H), 2.08-1.77 (m, 7H), 1.68-1.56 (m, 2H), 1.54-1.32 (m, 10H), 1.29-0.97 (m, 10H), 0.97-0.89 (m, 6H), 0.88-0.74 (m, 5H), 0.68 (s, 3H).

[1573] LCMS Rt=1.353 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.45O [M+H−H2O]+ 385, found 385.

[1574] SFC Rt=6.041 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, purity: 95%.

Synthesis to Determine Stereochemistry (8584 and 9142)

[1575] ##STR00460## ##STR00461##

[1576] To a solution of M-2-11_7 (400 mg, 0.849 mmol) in anhydrous THF (3 mL) was added n-BuLi (1.01 mL, 2.54 mmol, 2.5 M in n-hexane) drop-wise at −70° C. under N.sub.2. After stirring at −70° C. for 30 mins, a solution of (R)-2-ethyloxirane (91.5 mg, 1.27 mmol) in anhydrous THF (0.5 mL) was added drop-wise at −70° C. The reaction mixture was stirred at −70° C. for another 1 h and then stirred at 25° C. (room temperature) for 12 h. After heating at 60° C. for 2 h, the reaction was quenched by saturated aqueous NH.sub.4Cl (50 mL). The aqueous phase was extracted with EtOAc (3×50 mL). The combine organic phase was washed with saturated brine (2×50 mL). dried over anhydrous Na.sub.2SO.sub.4. filtered and concentrated in vacuum to give ST-200-94-10_1 (0.4 g, crude) as an oil, which was used directly of the next step.

[1577] To a solution of ST-200-094-010_1 (0.4 g, crude) in MeOH (50 mL) was added Mg powder (883 mg, 36.8 mmol) and NiCl.sub.2 (20 mg) at 25° C. under N.sub.2. After stirring at 60° C. for 1 h, the reaction mixture was quenched with HCl (100 mL, 1 M) until the reaction became clear. The aqueous phase was extracted with EtOAc (3×80 mL). The combined organic phase was washed with saturated NaHCO.sub.3.aq (2×50 mL), washed saturated brine (2×50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=10/1 to 8/1) to afford 8584 (180 mg, 61%) as a solid.

[1578] The ST-200-094-010 (180 mg, 0.447 mmol) was purified by SFC (Column: AD (250 mm*30 mm, 5 um), Condition: 0.1% NH.sub.3H.sub.2O IPA, Begin B: 40%, End B: 40%) to afford 8584 (120 mg, 67%) as a solid.

[1579] .sup.1H NMR (400 MHz, CDCl.sub.3) δ.sub.H 5.40-5.37 (m, 1H), 3.48-3.46 (m, 1H), 2.25-2.21 (m, 1H), 2.05-1.74 (m, 7H), 1.65-1.40 (m, 13H), 1.38-1.07 (m, 11H), 1.06-0.96 (m, 6H), 0.85 (s, 3H), 0.68 (s, 3H).

[1580] LCMS=1.277 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.45O [M+H−H.sub.2O].sup.+ 385, found 385.

[1581] SFC Rt=5.736 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 99.5% de.

[1582] A solution of 8584 (88 mg, 0.2185 mmol) and Pd(OH).sub.2 (80 mg) in MeOH (10 mL) was hydrogenated under 50 psi of hydrogen at 50° C. for 12 hours. The reaction mixture was filtered through a pad of celite and the filter cake was washed with THF (3×100 mL). The filter liquor was concentrated in vacuum. The residue was purified by flash column (10-25% of EtOAc in PE) to give 9142 (27 mg, 31%) as a solid.

[1583] .sup.1H NMR (400 MHz, CDCl.sub.3) δ.sub.H 3.50-3.41 (m, 1H), 1.99-1.91 (m, 1H), 1.87-1.74 (m, 3H), 1.70-1.60 (m, 3H), 1.53-1.19 (m, 12H), 1.18-0.97 (m, 11H), 0.96-0.78 (m, 12H), 0.75-0.54 (m, 5H).

[1584] LCMS Rt=1.292 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.45 [M+H−2H.sub.2O].sup.+ 369, found 369.

Example 86: Synthesis of 8689, 8602, and 8603

[1585] ##STR00462##

[1586] The experimental of intermediate 200-DA-C24_8_2 can be found in Example 15 and the synthesis of 200-N19-4_7 can be found in Example 83.

Synthesis of ST-200-6-18_1

[1587] ##STR00463##

[1588] A suspension of 200-N19-47 (600 mg, 1.29 mmol) in THF (8 mL) was added dropwise to a solution of n-BuLi (1.54 mL, 2.5 M in hexane, 3.87 mmol) in THF (2 mL) at −65° C. under N.sub.2. The mixture was stirred for 30 minutes at −65° C. Next, diisopropylamine (390 mg, 3.87 mmol) was added at −65° C., followed by adding 200-DA-C24_8_2 (387 mg, 3.87 mmol) dropwise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually and stirred at 25° C. for 16 hours. The reaction was quenched with saturated NH.sub.4Cl aq. (50 mL), extracted with EtOAc (3×50 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, concentrated and to give ST-200-6-18_1 (610 mg, crude) as an oil, which was used directly for the next step.

Synthesis of ST-200-6-18

[1589] ##STR00464##

[1590] Mg powder (1.01 g, 42.9 mmol) was added in 4 portions by stirring with a solution of ST-200-6-18_1 (610 mg, 1.06 mmol) and NiCl.sub.2 (13.6 mg, 0.106 mmol) in dry methanol (100 mL) under N.sub.2 at 50° C. The reaction mixture was stirred at 60° C. for 1 hour. The mixture was quenched with HCl (100 mL, 1 N) until the reaction became clear and extracted with EtOAc (3×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated. The residue was purified by flash column (0-30% of EtOAc in PE) to give 140 mg of product as a solid and 100 mg of impure product as a solid. The 140 mg product (0.325 mmol) was re-crystallized triturated from MeCN (10 mL) at 82° C. to give ST-200-6-18 (80 mg) as a solid.

[1591] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 4.05-3.95 (m, 1H), 3.95-3.83 (m, 1H), 3.72-3.62 (m, 1H), 3.55-3.45 (m, 1H), 2.50-2.40 (m, 1H), 2.05-1.55 (m, 13H), 1.50-1.32 (m, 7H), 1.32-1.03 (m, 9H), 1.03-0.75 (m, 8H), 0.68 (s, 3H).

[1592] LCMS Rt=1.086 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.28H.sub.45O.sub.2 [M+H−H.sub.2O].sup.+ 413, found 413.

Synthesis of ST-200-6-19 & ST-200-6-20

[1593] ##STR00465##

[1594] Stereochemistry was confirmed by Xray data for 8603.

[1595] 100 mg of ST-200-6-18 (0.232 mmol) was purified by SFC (Column: AD (150×4.6 mm, 3 um), Gradient: 5%-40% B (A: CO.sub.2 B: ethanol) Flow rate: 2.5 mL/min) to afford ST-200-6-19 (16.0 mg, 16%) as a solid and ST-200-6-20 (17.0 mg, 17%) as a solid.

[1596] 8602

[1597] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 4.05-3.95 (m, 1H), 3.95-3.83 (m, 1H), 3.72-3.62 (m, 1H), 3.55-3.48 (m, 1H), 2.30-2.15 (m, 1H), 2.05-1.55 (m, 12H), 1.50-1.32 (m, 7H), 1.32-1.03 (m, 10H), 1.03-0.70 (m, 8H), 0.68 (s, 3H).

[1598] LCMS Rt=1.088 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.28H.sub.45O.sub.2 [M+H−H.sub.2O].sup.+ 413, found 413.

[1599] 8603

[1600] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 4.05-3.95 (m, 1H), 3.95-3.83 (m, 1H), 3.72-3.62 (m, 1H), 3.55-3.48 (m, 1H), 2.50-2.40 (m, 1H), 2.05-1.55 (m, 13H), 1.50-1.32 (m, 6H), 1.32-1.03 (m, 10H), 1.00-0.75 (m, 8H), 0.68 (s, 3H).

[1601] LCMS Rt=1.084 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.28H.sub.45O.sub.2 [M+H−H.sub.2O].sup.+ 413, found 413.

Example 87: Synthesis of 8708

[1602] ##STR00466##

[1603] The experimental of intermediate ST-200-N19-4_6 can be found in Example 83.

The Synthesis of the Epoxide

[1604] ##STR00467##

[1605] To a suspension of Me.sub.3SI (3.93 g, 19.3 mmol) in THF (20 mL) was added a solution of t-BuOK (3.33 g, 29.8 mmol) in THF (10 mL) under N.sub.2 at 15° C. The suspension was stirred at 15° C. for 30 mins. A solution of 200-DA-E31_1 (2 g, 14.9 mmol) in THF (5 mL) was added dropwise at 15° C. The mixture was stirred at 15° C. for 16 hrs. The mixture was quenched with Sat. NH.sub.4Cl (50 mL) and extracted with EtOAc (3×20 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, and concentrated to give 200-DA-E31_2 (1.8 g, 82%) as a solid.

[1606] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.72 (s, 2H), 2.20-1.85 (m, 8H).

Synthesis of ST-200-N19-L14_1

[1607] ##STR00468##

[1608] A suspension of ST-200-N19-4_6 (200 mg, 0.42 mmol) in THF (4 mL) was added dropwise to a solution of n-BuLi (0.4 mL, 2.5 M in hexane, 1.06 mmol) in THF (1 mL) at −70° C. under N.sub.2. The mixture was stirred for 30 min at −70° C. A solution of diisopropylamine (107 mg, 1.06 mmol) was added dropwise at −70° C., then a solution of 6,6-difluoro-1-oxaspiro[2.5]octane (94.4 mg, 0.64 mmol) was added dropwise at −70° C. The mixture was stirred for another 30 min and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 24 hours. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (5 mL), extracted with EtOAc (3×10 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-N19-L14_1 (290 mg, crude) as a solid, which was used directly for the next step.

Synthesis of ST-200-N19-L14

[1609] ##STR00469##

[1610] Mg powder (448 mg, 18.7 mmol) and NiCl.sub.2 (5 mg, 0.05 mmol) were added to a solution of ST-200-N19-L14_1 (290 mg, 0.47 mmol) in 50 mL of anhydrous MeOH by stirring under N.sub.2 at 60° C. The reaction mixture was quenched by 2 M HCl (50 mL) until solid was dissolved. The mixture was extracted with EtOAc (3×100 mL). The combined organic layer was washed with Sat. NaHCO.sub.3 (150 mL), brine (150 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-15% of EtOAc in PE) to give a solid (.sup.1HNMR showed the product contained 10% 22,23-olefin). To a solution of ST-200-N19-L14 in EtOAc (5 mL) was added Lindlar catalyst (100 mg) under N.sub.2. The suspension was degassed under vacuum and purged with H.sub.2 for three times. Then the solution was hydrogenated under 15 psi of hydrogen at 25° C. for 4 h. The mixture was filtered through a pad of celite and washed with EtOAc (3×10 mL). The filtrate was concentrated and concentrated to give impure ST-200-N19-L14 as a solid (.sup.1HNMR showed the product contained 8% 22,23-olefin). Lindlar catalyst (100 mg) was added to a solution of ST-200-N19-L14 in THF/MeOH (3/3 mL) under N.sub.2. The suspension was degassed under vacuum and purged with H.sub.2 for three times. Then the solution was hydrogenated under 15 psi of hydrogen at 25° C. for 4 h. The mixture was filtered through a pad of celite and washed with THF (3×10 mL). The filtrate was concentrated and triturated from PE (5 mL) and n-hexane (5 mL) at 25° C. to give ST-200-N19-L14 (19 mg, 31%) as a solid.

[1611] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.35 (m, 1H), 2.27-2.19 (m, 1H), 2.17-1.76 (m, 11H), 1.69-1.57 (m, 6H), 1.52-1.21 (m, 11H), 1.19-0.98 (m, 6H), 0.97-0.91 (m, 4H), 0.88-0.74 (m, 5H), 0.68 (s, 3H).

[1612] LCMS Rt=1.252 min in 2.0 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.30H.sub.47F.sub.2O [M+H−H.sub.2O].sup.+ 461, found 461.

Example 88: Synthesis of 8809

[1613] ##STR00470##

[1614] The experimental of intermediate ST-200-N19-L5S can be found in Example 85.

Synthesis of ST-200-N19-L5SA

[1615] ##STR00471##

[1616] Pd(OH).sub.2/C (100 mg) was added to ST-200-N19-L5S (45 mg, 0.11 mmol) in THF/MeOH (5 mL/5 mL). The mixture was degassed and back-filled with H.sub.2 3 times. Next, the reaction was stirred at 50° C. under 50 psi of H.sub.2 for 72 h. The reaction mixture was filtered through a pad of celite washed with THF (100 mL). The filtrate was concentrated to give ST-200-N19-L5SA as a solid, which was further triturated in n-hexane (3 mL) to give ST-200-N19-L5SA (5 mg, 11%) as a solid.

[1617] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.53-3.43 (m, 1H), 2.07-1.61 (m, 13H), 1.59-1.21 (m, 13H), 1.20-0.99 (m, 8H), 0.98-0.75 (m, 9H), 0.74-0.53 (m, 4H).

[1618] LCMS Rt=1.267 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.45 [M+H−2H.sub.2O].sup.+ 369, found 369.

Example 89: Synthesis of 8946 and 8963

[1619] ##STR00472##

[1620] The synthesis of M-2-11_7 can be found in Example 83. The synthesis of the epoxide can be found in Example 28.

Synthesis of M-2-11_8

[1621] ##STR00473##

[1622] n-BuLi (1.01 mL, 2.54 mmol, 2.5 M in hexane) was added to a solution of M-2-11_7 (400 mg, 0.849 mmol) in THF (5 mL) at −70° C. under N.sub.2. After stirring at −70° C. for 1 hrs, 6, 6-dimethyl-1-oxaspiro [2.5]octane (236 mg, 1.69 mmol) was added at −70° C. The reaction was allowed to 25° C. and stirred at 25° C. for 12 hours, quenched with NH.sub.4Cl (10 mL, sat. aq) and water (30 mL) and extracted with EtOAc (3×10 mL). The organic layers were concentrated in vacuum to give M-2-11_8 (400 mg, crude) as an oil, which was used directly for the next step.

Synthesis of M-2-11_9

[1623] ##STR00474##

[1624] Mg powder (792 mg, 32.6 mmol) was added to a solution of M-2-11_8 (400 mg, crude) in MeOH (80 mL) at 50° C. The mixture was stirred at 50° C. for 1 h. After cooling to 0° C., the mixture was quenched with HCl (50 mL, 2 M) until the reaction became clear and extracted with EtOAc (2×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue was purified by flash column (0-40% of EtOAc in PE) to give 150 mg of impure a solid, which was triturated from MeCN (3 mL) at 25° C. to give M-2-11_9 (120 mg, 40%) as a solid.

[1625] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.39-5.38 (s, 1H), 2.26-2.21 (m, 1H), 2.05-1.77 (m, 7H), 1.68-1.38 (m, 16H), 1.33-1.00 (m, 14H), 0.96-0.90 (m, 6H), 0.89-0.81 (m, 7H), 0.68 (s, 3H).

[1626] LCMS Rt=1.651 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.51 [M+H−2H.sub.2O].sup.+ 435, found 435.

Synthesis of M-2-11

[1627] ##STR00475##

[1628] Dry Pd (OH).sub.2 (118 mg, 0.845 mmol) was added to a solution of M-2-11_9 (80 mg, 0.169 mmol) in THF (5 mL) under N.sub.2. The suspension was degassed under vacuum and purged with H.sub.2 3 times. The mixture was stirred under H.sub.2 (50 psi) at 50° C. for 12 hours to give a black suspension. The reaction mixture was filtered through a pad of celite and washed with THF (3×30 mL). The filtrate was concentrated in vacuum to get M-2-11 (20 mg, 25%) as a solid.

[1629] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.97-1.91 (s, 1H), 1.88-1.76 (m, 4H), 1.69-1.60 (m, 3H), 1.59-1.46 (m, 15H), 1.31-1.06 (m, 14H), 0.94-0.86 (m, 15H), 0.69-0.63 (m, 4H).

[1630] LCMS Rt=1.718 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.32H.sub.52 [M+H−2H.sub.2O].sup.+ 437, found 437.

Example 90: Synthesis of 9062

[1631] ##STR00476##

Synthesis for the Experimental of Intermediate 200-N19-M22_6 (or M-4-14-2)

Synthesis of 200-N19-3_1

[1632] ##STR00477##

[1633] t-BuOH (1.7 L) was charged into a three-neck round bottom flask under N.sub.2 at 35° C. and stirred for 10 mins. t-BuOK (292 g, 2.61 mol) was added to the mixture and stirred until the reaction became clear. After that, ST-310-B9_1 (65 g, 238 mmol) was added to the above mixture and stirred for 1.5 h at 35° C. under N.sub.2. The reaction mixture was poured into 10% aqueous acetic acid (2 L) and stirred for 30 mins, during which the temperature was maintained below 10° C. Then the mixture was treated with water (1.5 L) and the pH was adjusted to 7˜8 with NaHCO.sub.3 and stirred for 30 mins. The aqueous phase was extracted with MTBE (3 L). The organic layer was separated, washed with brine (3×1 L), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated below 35° C. to give ST-200-N19-3_1 (65 g, crude) as an oil. The crude residue was used directly for the next step.

Synthesis of 200-N19-M22_1

[1634] ##STR00478##

[1635] To a solution of 2,6-di-tert-butyl-4-methylphenol (340 g, 1.54 mol) in toluene (700 mL) was added drop-wise AlMe.sub.3 (385 mL, 770 mmol, 2 M in toluene) at 0° C. The mixture was stirred at 25° C. for 1 h and used directly as MAD solution. A solution of 200-N19-3_1 (60 g, 220 mmol) in anhydrous toluene (200 mL) and anhydrous DCM (200 mL) was added to MAD solution at −70° C. over a period of 30 mins under N.sub.2. The reaction mixture stirring at −70° C. for 1 h. Then MeMgBr (220 mL, 660 mmol, 3M in ethyl ether) was added drop wise at −70° C. and stirred for 1 h. The reaction was poured into saturated aqueous citric acid (2 L) at 0° C. and stirring for 30 min, extracted with EtOAc (2×1 L). The combined organic phase was washed with saturated brine (2×1 L), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=10/1 to 5/1) to afford 200-N19-M22_1 (33 g, 52%) as a solid.

[1636] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.46-5.42 (m, 1H), 2.25-2.40 (m, 1H), 2.21-1.60 (m, 13H), 1.35-1.21 (m, 4H), 1.13 (s, 3H), 0.98-0.83 (m, 6H).

Synthesis of M-4-14_1

[1637] ##STR00479##

[1638] To a suspension of Ph.sub.3PEtBr (102 g, 277 mmol) in Anhydrous THF (500 mL) was added t-BuOK (31.0 g, 277 mmol) in one portion at 25° C. under N.sub.2. The reaction mixture turned to dark red. After stirring at 25° C. for 30 min, 200-N19-M22_1 (20 g, 69.3 mmol) was added and stirred for 2 h at 25° C. The reaction was quenched with aq. NH.sub.4Cl (800 mL) at 0° C., extracted with EtOAc (2×500 mL). The combined organic phase was washed with brine (2×500 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=10/1 to 5/1) to afford M-4-14_1 (15 g, 72%) as a solid.

[1639] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.43-5.40 (m, 1H), 5.16-5.10 (m, 1H), 2.41-2.33 (m, 1H), 2.28-1.86 (m, 8H), 1.78-1.71 (m, 1H), 1.69-1.50 (m, 11H), 1.41-1.10 (m, 6H), 0.94-0.81 (s, 3H).

Synthesis of M-4-14_2

[1640] ##STR00480##

[1641] To a solution of M-4-14_1 (30 g, 99.8 mmol) in anhydrous THF (500 mL) was added 9-BNN dimer (66.9 g, 299 mmol) and stirred for 30 mins at 0° C. under N.sub.2. The reaction mixture was warmed to 50° C. and stirred for 1 h. after cooled to 0° C. and EtOH (100 mL) was added. NaOH. aq (99.8 mL, 5M, 499 mmol) was added very slowly. H.sub.2O.sub.2 (53.0 g, 499 mmol, 30% in water) was added slowly and the inner temperature was maintained below 30° C. The mixture was warmed to 50° C. and stirred for 1 h. The reaction mixture was cooled and ice-water (1 L) was added and stirred 30 min. filtered and concentrated in vacuum to give M-4-11_2 (30 g, crude) as a solid. The crude material was used directly for the next step.

[1642] The synthesis of the tosylate can be found in Example 30.

Synthesis of 200-N19-M22_7

[1643] ##STR00481##

[1644] TEA (21 mL) and TsCl (16.0 g, 84.0 mmol) were added to a solution of 200-N19-M22_6 (7 g, 21.0 mmol) in DCM (150 mL) at 25° C. The mixture was stirred at 40° C. for 12 h. To the reaction was added water (200 mL). The aqueous phase was extracted with DCM (2×200 mL). The combined organic phase was washed with saturated brine (2×200 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated was purified by silica gel chromatography (PE/EtOAc=10/1 to 8/1) to afford 200-N19-M22_7 (10 g, 98%) as an oil.

[1645] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.79-7.76 (m, 2H), 7.34-7.26 (m, 2H), 5.38-5.29 (m, 1H), 3.95-3.93 (m, 1H), 3.74-3.73 (m, 1H), 2.44 (s, 3H), 2.19-2.14 (m, 1H), 2.09-1.97 (m, 3H), 1.92-1.44 (m, 14H), 1.29-1.08 (m, 6H), 1.05-0.88 (m, 5H), 0.63 (s, 3H).

Synthesis of 200-N19-M22_8

[1646] ##STR00482##

[1647] KI (16.9 g, 102 mmol) was added to a solution of 200-N19-M22_7 (10 g, 20.5 mmol) in DMF (100 mL) at 25° C. under N.sub.2. The mixture was stirred at 50° C. for 12 h under N.sub.2. The residue was poured into ice-water (300 mL) and stirred for 20 min. The aqueous phase was extracted with EtOAc (2×200 mL). The combined organic phase was washed with saturated brine (2×200 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give 200-N19-M22_8 (8 g, 88%) as a solid.

[1648] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.41-5.39 (m, 1H), 3.40-3.28 (m, 1H), 3.23-3.13 (m, 1H), 2.19-2.13 (m, 1H), 2.10-1.71 (m, 9H), 1.63-1.34 (m, 7H), 1.16-0.98 (m, 10H), 0.96-0.77 (m, 2H), 0.72 (s, 3H).

Synthesis of 200-N19-M22

[1649] ##STR00483##

[1650] PhSO.sub.2Na (9.27 g, 56.5 mmol) was added to a solution of 200-N19-M22_8 (5 g, 11.3 mmol) in DMF (50 mL) and stirred at 50° C. for 6 h. The reaction mixture was cooled to 25° C. and added water (200 mL). The aqueous phase was extracted with EtOAc (2×100 mL). The combined organic phase was washed with saturated brine (2×100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by a silica gel column (PE/EtOAc=8/1-5/1) to give 200-N19-M22 (4.0 g) as a solid. The 200-N19-M22 (4.0 g, 8.75 mmol) was re-crystallized from MeCN (50 mL) at 82° C. reflux for 1 hrs. The mixture stirred was cooled to 25° C. (room temperature). The suspension was filtration in vacuum to get 200-N19-M22 (3.5 g, 68%) as a solid.

[1651] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.94-7.88 (m, 2H), 7.66-7.54 (m, 3H), 5.39-5.37 (m, 1H), 3.17-3.13 (m, 1H), 2.88-2.81 (m, 1H), 2.18-2.12 (m, 1H), 2.11-1.47 (m, 15H), 1.28-1.08 (m, 8H), 1.07-0.74 (m, 5H), 0.65 (s, 3H).

Synthesis of ST-200-087-001_1

[1652] ##STR00484##

[1653] n-BuLi (1.22 mL, 3.06 mmol, 2.5M in n-hexane) was added dropwise to a solution of 200-N19-M22 (400 mg, 0.876 mmol) in anhydrous THF (3.5 mL) at −70° C. under N.sub.2. After stirring at −70° C. for 30 mins, a solution of (S)-3,3,3-trifluoro-2-hydroxy-2-methylpropyl 4-methylbenzenesulfonate (390 mg, 1.31 mmol) in anhydrous THF (0.5 mL) was added drop-wise at −70° C. and stirred for another 1 h. The reaction mixture was stirred at 25° C. (room temperature) for 12 h. The reaction was quenched by saturated Cl. aq (20 mL). The aqueous phase was extracted with EtOAc (3×50 mL). The combine organic phase was washed whit saturated brine (2×50 mL), drive over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give ST-200-087-001_1 (0.4 g, crude) as a solid, which was used directly.

Synthesis of ST-200-087-001

[1654] ##STR00485##

[1655] Mg powder (986 mg, 41.1 mmol) and NiCl.sub.2 (20 mg) were added to a solution of ST-200-087-001_1 (0.4 g, crude) in MeOH (50 mL) at 25° C. under N.sub.2. After stirring at 50° C. for 1 h under N.sub.2, the reaction mixture was quenched with HCl (100 mL, 1 M) until the reaction became clear. The aqueous phase was extracted with EtOAc (3×80 mL). The combined organic phase was washed with saturated brine (2×50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=8/1 to 5/1) to afford ST-200-087-001 (63 mg, 20.7%) as a solid and ST-200-087-001 (80 mg, impure) as a solid.

[1656] Lindlar catalyst (300 mg) was added to a solution of ST-200-087-001 (143 mg, 0.323 mmol) in anhydrous THF (2 mL) under N.sub.2. The suspension was degassed under vacuum and purged with H.sub.2 for three times. The mixture was stirred under H.sub.2 (15 psi) at 25° C. for 4 hours to give a black suspension, The reaction mixture was filtered through a pad of Celite and washed with EtOAc (2×30 mL). The filtrate was concentrated in vacuum to give ST-200-087-001 (80 mg, 56%) as a solid.

[1657] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.41-5.39 (m, 1H), 2.19-2.14 (m, 1H), 2.10-1.61 (m, 11H), 1.54-1.16 (m, 13H), 1.14-0.73 (m, 12H), 0.69 (s, 3H).

[1658] LCMS Rt=1.217 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.26H.sub.40F.sub.3O [M+H−H.sub.2O].sup.+ 425, found 425.

Example 91: Synthesis of 9142

[1659] ##STR00486## ##STR00487##

[1660] The experimental of intermediate M-2-11_7 can be found in Example 83.

[1661] See Example 85 for stereochemistry synthesis.

Synthesis of ST-200-091-002_1

[1662] ##STR00488##

[1663] To a solution of R, R-cat (166 mg, 0.276 mmol) in toluene (5 mL) was added AcOH (173 mg, 2.89 mmol). The mixture was stirred at 25° C. open to air for 30 min and concentrated in vacuo to leave a crude solid. The resulting catalyst residue was dissolved in 2-ethyloxirane (10 g, 138 mmol) at 25° C. The reaction flask was cooled to 0° C. and treated with H.sub.2O (1.36 g, 75.9 mmol) dropwise over 5 min. The reaction was allowed to warm to 25° C. and stir 24 hrs and the reaction mixture was distilled to give (R)-2-ethyloxirane (4.4 g, 61.0 mmol).

[1664] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.91-2.88 (m, 1H), 2.76-2.71 (m, 1H), 2.49-2.47 (m, 1H), 1.62-1.54 (m, 2H), 1.03-0.97 (m, 3H).

[1665] The ee value of the epoxide was determined as following

##STR00489##

[1666] To a solution of (2R)-2-ethyloxirane (100 mg, 1.38 mmol) and naphthalene-2-thiol (221 mg, 1.38 mmol) in methanol (10 mL) was added triethylamine (139 mg, 1.38 mmol). The mixture was stirred at 30° C. for 16 hrs. The reaction mixture was used directly to determine ee % without any treatment. The ee % was determined to be 93.6%.

[1667] SFC Rt=5.287 min in 10 min chromatography, AD-3_IPA(DEA)_5_40_2.5ML, 93.6% ee.

Synthesis of ST-200-094-010_1

[1668] ##STR00490##

[1669] To a solution of M-2-11_7 (400 mg, 0.849 mmol) in anhydrous THF (3 mL) was added n-BuLi (1.01 mL, 2.54 mmol, 2.5 M in n-hexane) drop-wise at −70° C. under N.sub.2. After stirring at −70° C. for 30 mins, a solution of (R)-2-ethyloxirane (91.5 mg, 1.27 mmol) in anhydrous THF (0.5 mL) was added drop-wise at −70° C. The reaction mixture was stirred at −70° C. for another 1 h and then stirred at 25° C. (room temperature) for 12 h. After heating at 60° C. for 2 h, the reaction was quenched by saturated aqueous NH.sub.4Cl (50 mL). The aqueous phase was extracted with EtOAc (3×50 mL). The combine organic phase was washed with saturated brine (2×50 mL). dried over anhydrous Na.sub.2SO.sub.4. filtered and concentrated in vacuum to give ST-200-94-10_1 (0.4 g, crude) as an oil, which was used directly of the next step.

Synthesis of ST-200-094-010

[1670] ##STR00491##

[1671] To a solution of ST-200-094-010_1 (0.4 g, crude) in MeOH (50 mL) was added Mg powder (883 mg, 36.8 mmol) and NiCl.sub.2 (20 mg) at 25° C. under N.sub.2. After stirring at 60° C. for 1 h, the reaction mixture was quenched with HCl (100 mL, 1 M) until the reaction became clear. The aqueous phase was extracted with EtOAc (3×80 mL). The combined organic phase was washed with saturated NaHCO.sub.3.aq (2×50 mL), washed saturated brine (2×50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=10/1 to 8/1) to afford ST-200-094-010 (180 mg, 61%) as a solid. The ST-200-094-010 (180 mg, 0.447 mmol) was purified by SFC (Column: AD (250 mm*30 mm, 5 um), Condition: 0.1% NH.sub.3H.sub.2O IPA, Begin B: 40%, End B: 40%) to afford ST-200-094-010 (120 mg, 67%) as a solid.

[1672] .sup.1H NMR (400 MHz, CDCl.sub.3) δ.sub.H 5.40-5.37 (m, 1H), 3.48-3.46 (m, 1H), 2.25-2.21 (m, 1H), 2.05-1.74 (m, 7H), 1.65-1.40 (m, 13H), 1.38-1.07 (m, 11H), 1.06-0.96 (m, 6H), 0.85 (s, 3H), 0.68 (s, 3H).

[1673] LCMS=1.277 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.45O [M+H−H.sub.2O].sup.+ 385, found 385.

[1674] SFC Rt=5.736 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 99.5% de.

Synthesis of ST-200-094-012

[1675] ##STR00492##

[1676] A solution of ST-200-094-010 (88 mg, 0.2185 mmol) and Pd(OH).sub.2 (80 mg) in MeOH (10 mL) was hydrogenated under 50 psi of hydrogen at 50° C. for 12 hours. The reaction mixture was filtered through a pad of celite and the filter cake was washed with THF (3×100 mL). The filter liquor was concentrated in vacuum. The residue was purified by flash column (10-25% of EtOAc in PE) to give ST-200-094-012 (27 mg, 31%) as a solid.

[1677] .sup.1H NMR (400 MHz, CDCl.sub.3) δ.sub.H 3.50-3.41 (m, 1H), 1.99-1.91 (m, 1H), 1.87-1.74 (m, 3H), 1.70-1.60 (m, 3H), 1.53-1.19 (m, 12H), 1.18-0.97 (m, 11H), 0.96-0.78 (m, 12H), 0.75-0.54 (m, 5H).

[1678] LCMS Rt=1.292 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.45 [M+H−2H.sub.2O].sup.+ 369, found 369.

Example 93. Biological Data

[1679] The experiments were conducted as described in Example 2 and the results are provided in Table 2-61.

TABLE-US-00005 TABLE 2-61 Avg Avg Avg Avg EC50 Emax EC50 Emax Compound 2A (nM) 2A (%) 2B (nM) 2B (%) [00493] 376 >10000 49.9 >10000 59.8 [00494] 1A75 671.1 457.2 439.6 308.0 [00495] 1B75 164.9 278.8 183.4 233.1 [00496] 476 406.0 811.2 189.5 399.2 [00497] 175 574.1 366.7 287.5 279.0 [00498] 679 5113.4 104.9 280.0 171.5 [00499] 4A77 427.2 382.5 285.2 253.7 [00500] 4B77 466.8 466.6 298.2 320.5 [00501] 780 340.9 132.1 263.2 169.7 [00502] 8127 543.0 135.7 571.2 210.7 [00503] 8245 524.5 73.0 267.6 119.5 [00504] 8361 87.0 252.0 136.3 308.8 [00505] 8462 170.0 273.2 96.2 281.0 [00506] 8463 102.5 291.3 85.1 324.2 [00507] 8564 468.8 252.5 489.5 310.5 [00508] 8378 123.9 195.0 208.3 258.6 [00509] 8379 73.4 251.8 78.1 358.6 [00510] 8584 332.2 284.3 244.5 363.1 [00511] 8585 286.2 194.4 236.7 232.9 [00512] 8689 168.6 319.6 174.8 438.6 [00513] 8602 1047.1 271.7 433.7 262.3 [00514] 8603 134.9 293.5 75.1 270.6 [00515] 8708 697.2 117.4 552.7 243.2 [00516] 8809 532.9 73.6 160.4 78.2 [00517] 8946 1596.2 262.8 1352.5 231.4 [00518] 8963 >10000 8.4 >10000 55.9 [00519] 9062 34.5 66.5 179.0 52.2 [00520] 9142 >10000 41.6 70.4 44.8

Example 94. Synthesis of Compound 194

[1680] ##STR00521##

Step 1

[1681] t-BuOH (350 mL) was charged into a three-neck round bottom flask under nitrogen at 35° C. and stirred under nitrogen gas bubbling for 10 mins. t-BuOK (90.5 g, 807 mmol) was added to the mixture and stirred under nitrogen gas bubbling for 15 mins. A194 (20 g, 73.4 mmol) was added to the above mixture and stirred under nitrogen gas bubbling at 35° C. for 1.5 hrs. The reaction mixture was poured into 10% aqueous acetic acid (500 mL) and stirred for 15 mins below 35° C. Water (500 mL) was added to the reaction and stirred for 30 mins. The pH of the mixture was adjusted to 7-8 with sodium bicarbonate (500 ml). The mixture was stirred for 30 mins. The mixture was extracted with PE (2×500 mL). The organic layer was separated, washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered and concentrated below 35° C. to give A294 (17 g, crude) as an oil. The crude residue was used directly for the next step.

Step 2

[1682] To a solution of 2, 6-di-tert-butyl-4-methylphenol (100 g, 453 mmol) in toluene (300 ml) was added drop-wise AlMe.sub.3 (113 mL, 226 mmol, 2 M in toluene) at 0° C. The mixture was stirred at 25° C. for 1 h. A solution of A294 (10 g, 36.7 mmol) in toluene (50 mL) was added dropwise at −70° C. and after stirring at −70° C. for 1 h, EtMgBr (36.6 ml, 110 mmol, 3M in ethyl ether) was added drop wise at −70° C. The resulting solution was stirred at −70° C. for 1 hr. The reaction mixture was quenched by saturated citric acid (400 ml) at −70° C. After stirring at 25° C. for 10 min, the resulting mixture was filtered and washed with EtOAc (2×200 ml). The combined organic layer was separated, washed with brine (2×200 ml), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue was purified by silica gel chromatography (PE/EtOAc=10/1 to 5/1) to get A394 (7.6 g, impure) as a solid.

[1683] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.40 (m, 1H), 2.51-2.38 (m, 1H), 2.49-2.21 (m, 1H), 2.14-1.88 (m, 5H), 1.86-1.77 (m, 2H), 1.73-1.38 (m, 8H), 1.34-1.22 (m, 4H), 0.95-0.81 (m, 8H).

Step 3

[1684] To a suspension of PPh.sub.3EtBr (37.1 g, 100 mmol) in THF (200 mL) under N.sub.2 was added t-BuOK (11.2 g, 100 mmol) at 40° C. After stirring at 20° C. for 10 min, A394 (7.6 g, 25.1 mmol) was added. The reaction mixture was stirred at 40° C. for 1 h. The reaction was quenched with saturated aqueous NH.sub.4Cl (200 mL) at 0° C. and extracted with EtOAc (3×200 mL). The combined organic phase was washed with brine (200 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash chromatography (0%-30% of EtOAc in PE) to afford A494 (5 g, 63%) as a solid.

[1685] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.35 (m, 1H), 5.20-5.00 (m, 1H), 2.41-2.30 (m, 1H), 2.29-2.12 (m, 3H), 2.09-1.76 (m, 6H), 1.69-1.38 (m, 15H), 1.35-0.94 (m, 7H).

Step 4

[1686] To a solution of A494 (2 g, 6.35 mmol) in THF (20 mL) was added 9-BBN dimer (3.09 g, 12.7 mmol) at 0° C. under N.sub.2. The solution was stirred at 60° C. for 1 h. After cooling to 0° C., a solution of EtOH (20 ml) and NaOH (12.7 ml, 5M, 63.5 mmol) was added very slowly. After addition, H.sub.2O.sub.2 (2.15 mg, 6.35 mmol, 30% in water) was added slowly and the inner temperature was maintained below 10° C. The mixture was stirred at 60° C. under N.sub.2 for 1 hour. The mixture was re-cooled to 30° C., water (100 mL) was added to the solution and extracted with EtOAc (100 mL). The organic layer was washed with brine (2×100 mL) and then the combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, and purified by silica gel chromatography (PE/EtOAc=2/1) to afford impure A594 (1.6 g) as a solid.

[1687] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.35 (m, 1H), 3.75-3.62 (m, 1H), 2.28-2.19 (m, 1H), 2.10-1.75 (m, 7H), 1.71-0.97 (m, 19H), 0.92-0.75 (m, 4H), 0.68 (s, 3H).

Step 5

[1688] To a solution of A594 (1.6 g, 4.81 mmol) in DCM (20 mL) was added silica gel (2 g) and PCC (2.07 g, 9.62 mmol). The mixture was stirred at 25° C. for 3 hrs. To the mixture was added PE (50 mL) and the mixture was filtered though a pad of silica gel and the solid was washed with PE/DCM (30 mL/30 mL). The mixture was filtered and filtrate was concentrated in vacuum. The residue was purified by silica gel chromatography (PE/EtOAc=10/1 to 5/1) to afford impure A094 (1.2 g) as a solid, which was re-crystallized from MeCN (10 mL) at reflux (82° C.) to give A694 (1.0 g, 84%) as a solid.

[1689] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 2.61-2.45 (m, 1H), 2.30-2.10 (m, 5H), 2.00-1.75 (m, 6H), 1.70-1.10 (m, 14H), 0.90-0.75 (m, 4H); 0.633 (s, 3H).

[1690] LCMS Rt=1.058 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.22H.sub.33O [M+H−H.sub.2O].sup.+ 313, found 313.

Step 6

[1691] To a suspension of Ph.sub.3PMeBr (11.1 g, 31.4 mmol) in THF (50 mL) under N.sub.2 was added t-BuOK (3.51 g, 31.4 mmol) at 40° C. After stirring at 25° C. for 10 min, A6 (2.6 g, 7.86 mmol) was added. The reaction mixture was stirred at 40° C. for 1 h. The reaction was quenched with ag. NH.sub.4Cl (100 mL) at 0° C., extracted with EtOAc (2×100 mL). The combined organic phase was washed with brine (2×100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash chromatography (0%˜30%, EtOAc in PE) to afford A794 (2.4 g, 93%) as a solid.

[1692] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.35 (m, 1H), 4.86-4.83 (m, 1H), 8.70-4.65 (m, 1H), 2.27-2.20 (m, 1H), 2.10-1.90 (m, 4H), 1.89-1.50 (m, 11H), 1.49-1.30 (m, 3H), 1.28-1.00 (m, 6H), 0.80-0.60 (m, 5H), 0.59 (s, 3H).

Step 7

[1693] To a solution of A794 (2.4 g, 7.30 mmol) in THF (40 mL) was added 9-BBN dimer (4.44 g, 18.2 mmol) at 0° C. under N.sub.2. The solution was stirred at 60° C. for 1 h. After cooling to 0° C., a solution of EtOH (30 ml) and NaOH (14.5 mL, 5M, 73.0 mmol) was added very slowly. After addition, H.sub.2O.sub.2 (7.29 mL, 73.0 mmol, 30% in water) was added slowly and the inner temperature was maintained below 10° C. The mixture was stirred at 60° C. under N.sub.2 for 1 hour. The mixture was re-cooled to 30° C. and water (100 mL) was added to the solution with EtOH (50 mL). A precipitate appeared, which was collected by filtration and concentrated in vacuum to give A894 (1.8 g, 71%) as a solid.

[1694] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.35 (m, 1H), 3.69-3.60 (m, 1H), 3.40-3.30 (m, 1H), 2.25-2.00 (m, 1H), 2.08-1.75 (m, 7H), 1.68-1.60 (m, 2H), 1.55-1.38 (m, 5H). 1.36-1.09 (m, 8H), 1.08-0.93 (m, 4H), 0.89-0.76 (m, 5H), 0.70 (s, 3H).

Step 8

[1695] To a solution of A894 (1 g, 2.88 mmol) in chloroform (5.5 mL) and pyridine (3.5 mL) was added TsCl (1.42 g, 7.48 mmol) at 25° C. The mixture was stirred at 25° C. for 2 hrs. The reaction mixture was concentrated under vacuum to remove most of the chloroform. To the obtained pyridine mixture was added water (50 mL). A solid was produced, which was collected by filtration and washed with water (5×50 mL). The solid was dissolved in DCM (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give A994 (1.2 g, crude) as an oil, which was used directly for the next step.

Step 9

[1696] To a solution of A994 (800 mg, 1.55 mmol) in DMF (6 mL) was added KI (1.23 g, 7.44 mmol) at 25° C. The mixture was stirred at 50° C. for 1 hour. The reaction mixture was poured into water (50 mL) with PE (30 mL). The organic phase was washed with brine (2×30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to give A1094 (700 mg, 96%) as an oil.

[1697] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.45-5.35 (m, 1H), 3.36-3.30 (m, 1H), 3.20-3.10 (m, 1H), 2.30-2.20 (m, 1H), 2.07-1.70 (m, 7H), 1.68-1.60 (m, 2H), 1.32-1.14 (m, 8H), 1.13-0.94 (m, 6H), 0.93-0.74 (m, 7H), 0.72 (s, 3H).

Step 10

[1698] To a solution of PhSO.sub.2Me (449 mg, 2.88 mmol) in THF (10 mL) was added n-BuLi (1.04 mL, 2.62 mmol, 2.5 M in hexane) at −70° C. under N.sub.2. The mixture was warmed to 0° C. A suspension of A1094 (600 mg, 1.31 mmol) in THF (10 mL) was added drop-wise at 0° C. After addition, the reaction was allowed to 25° C. The reaction mixture was stirred at 25° C. for 1 hour. The reaction was quenched with saturated aqueous NH.sub.4Cl (30 mL). To the suspension was added water (100 mL) and extracted with EtOAc (3×50 mL). The combined organic phase was concentrated to give a residue which was purified by silica gel chromatography (PE/EtOAc=6/1) to give compound A1194 (1.5 g, impure, containing PhSO.sub.2Me) as an oil. The oil was further purified by silica gel chromatography (DCM/Acetone=50/1) to give A1194 (400 mg, 63%) as an oil.

[1699] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.90 (d, J=7.6 Hz, 2H), 7.69-7.62 (m, 1H), 7.60-7.51 (m, 2H), 5.40-5.35 (m, 1H), 3.17-3.07 (m, 1H), 3.04-2.93 (m, 1H), 2.22 (dd, J=2.4, 12.8 Hz, 1H), 2.10-1.64 (m, 9H), 1.57-1.35 (m, 7H), 1.31-1.11 (m, 5H), 1.10-0.92 (m, 3H), 0.90-0.82 (m, 7H), 0.81-0.71 (m, 1H), 0.64 (s, 3H).

Step 11

[1700] To a solution of n-BuLi (0.468 mL, 2.5 M in hexane, 1.17 mmol) in THF (0.5 mL) at −65° C. under N.sub.2 was added a suspension of A1194 (200 mg, 0.39 mmol) in THF (2.5 mL) dropwise. The mixture was stirred for 30 minutes at −65° C. Then 2,2-dimethyloxirane (42.1 mg, 0.585 mmol) was added dropwise at −65° C. The mixture was stirred for another 30 minutes and then gradually warmed to 25° C. The reaction mixture was stirred at 25° C. for 16 hours. The reaction mixture was quenched with saturated aqueous NH.sub.4Cl (30 mL) and extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give A1294 (210 mg) as an oil, which was used directly for the next step.

Step 12

[1701] To a solution of A1294 (210 mg, 0.3771 mmol) and nickel(II) chloride (5.02 mg, 0.03875 mmol) in dry methanol (100 mL) was added magnesium powder (372 mg, 15.5 mol) in 3 portions under N.sub.2 with stirring at 50° C. to initiate continuous hydrogen generation. The reaction mixture was stirred at 60° C. for 1 hour. The reaction mixture was quenched with 2 M HCl (30 mL) which was added dropwise at 10° C. until the solid was dissolved. After extracting with EtOAc (2×50 mL), the combined organic layer was washed with saturated aqueous NaHCO.sub.3 (30 mL), brine (30 mL) then dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a solid, which was purified by silica gel chromatography (PE/EtOAc=10/1) to give 100 mg of impure product, which was triturated from MeCN (5 mL) to give Compound 194 (16.5 mg, 11%) as a solid along with 80 mg of impure product.

[1702] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.36 (m, 1H), 2.23 (dd, 7=2.8, 13.2 Hz, 1H), 2.08-1.89 (m, 4H), 1.85-1.76 (m, 3H), 1.67-1.58 (m, 2H), 1.53-1.32 (m, 9H), 1.32-1.23 (m, 4H), 1.21 (s, 6H), 1.20-1.15 (m, 3H), 1.15-0.97 (m, 4H), 0.93 (d, J=6.8 Hz, 3H), 0.86 (t, J=7.2 Hz, 4H), 0.83-0.74 (m, 1H), 0.68 (s, 3H).

[1703] LCMS Rt=1.355 min in 2.0 min chromatography, 30-90AB, purity 94.6% (ELSD), MS ESI calcd. for C.sub.28H.sub.47O [M+H−H.sub.2O].sup.+ 399, found 399.

Example 95: Synthesis of 9567

[1704] ##STR00522##

[1705] The synthesis of 200-N19-4_7 can be found in Example 94.

Synthesis of ST-200-52-9_1

[1706] ##STR00523##

[1707] n-BuLi (0.99 mL, 2.5 M, 2.47 mmol) was added to a solution of diisopropylamine (0.38 mL, 2.66 mmol) in THF (1 mL) under N.sub.2 at −70° C. The resulting mixture was warmed to 25° C. and stirred at 25° C. for 30 min. After re-cooling to −70° C., a solution of 200-N19-4_7 (0.3 g, 0.62 mmol) in THF (3 mL) was added at −70° C. After stirring at −70° C. for 1 hour, (S)-2-(trifluoromethyl)oxirane (69.3 mg, 0.62 mmol) was added at −70° C. The reaction mixture was warmed to 25° C. and stirred at 25° C. for 18 hours. The reaction mixture was quenched with saturated NH.sub.4Cl aqueous (6 mL) at 0° C. The mixture was extracted with EtOAc (2×8 mL). The combined organic phase was washed with brine (2×10 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a crude product, which was used directly for the next step.

Synthesis of ST-200-52-9

[1708] ##STR00524##

[1709] Mg powder (513 mg, 21.4 mmol) was added in 4 portions under N.sub.2 with stirring at 50° C. to a solution of ST-200-52-9_1 (320 mg, 0.536 mmol) and NiCl.sub.2 (6.91 mg, 0.054 mmol) in dry methanol (50 mL). After stirring at 60° C. for 1 hour, the mixture was quenched with HCl (50 mL, 1N) until the reaction became clear and extracted with EtOAc (3×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated. The residue was purified by flash column (0-15% of EtOAc in PE) to give ST-200-52-9 (11 mg, 5%) as a solid.

[1710] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.42-5.35 (m, 1H), 3.95-3.83 (m, 1H), 2.25-2.17 (m, 1H), 2.05-1.83 (m, 5H), 1.83-1.75 (m, 3H), 1.75-1.50 (m, 3H), 1.50-1.30 (m, 6H), 1.30-0.98 (m, 11H), 0.94 (s, 3H), 0.90-0.72 (m, 6H), 0.68 (s, 3H).

[1711] LCMS Rt=1.253 min in 2.0 min chromatography, 30-90AB_2MIN_E, 100% purity, MS ESI calcd. for C.sub.27H.sub.42F.sub.3O.sub.2 [M+H−H.sub.2O].sup.+ 439, found 439.

Example 96: Synthesis of 9670

[1712] ##STR00525##

[1713] The synthesis of ST-200-52-7 can be found in Example 94.

Synthesis of ST-200-52-6

[1714] ##STR00526##

[1715] Pd(OH).sub.2/C (dry, 200 mg) was added to a solution of ST-200-52-7 (200 mg, 0.479 mmol) in MeOH (30 mL). The mixture was hydrogenated under 50 psi at 50° C. for 48 hrs. The mixture was filtered, concentrated and purified by combi-flash (0-10% of EtOAc in PE) to give ST-200-52-6 (33 mg, 16%) as a solid.

[1716] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.99-1.90 (m, 1H), 1.88-1.75 (m, 3H), 1.69-1.56 (m, 4H), 1.55-1.52 (m, 4H), 1.48-1.32 (m, 5H), 1.31-1.23 (m, 2H), 1.22-1.15 (m, 9H), 1.13-0.99 (m, 9H), 0.95-0.78 (m, 8H), 0.74-0.53 (m, 5H).

[1717] LCMS Rt=1.299 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.28H.sub.47 [M+H−2H.sub.2O].sup.+ 383, found 383.

Example 97: Synthesis of 9792

[1718] ##STR00527##

[1719] The synthesis of ST-200-N19-4_7 can be found in Example 94.

Synthesis of ST-200-N19-K7R_1

[1720] ##STR00528##

[1721] n-BuLi (0.408 mL, 2.5 M, 1.02 mmol, 2.5 eq) was added to THF (0.5 mL) under N.sub.2 at −70° C. Next, a suspension of ST-200-N19-47 (200 mg, 0.412 mmol, 1.0 eq.) in THF (1.5 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, (R)-2-methyloxirane (35.8 mg, 0.618 mmol, 1.5 eq.) was added dropwise. Then reaction was stirred at 25° C. for 12 hrs. The reaction was quenched with sat. NH.sub.4Cl (30 mL), extracted with EtOAc (3×15 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude product (250 mg) as a solid, which was used directly for the next step.

Synthesis of ST-200-N19-K7R

[1722] ##STR00529##

[1723] Mg powder (441 mg, 18.4 mmol) was added in one portion to a solution of ST-200-N19-K7R_1 (250 mg, 0.46 mmol) in MeOH (30 mL) at 65° C. The mixture was stirred at 65° C. for 1 h. The mixture was quenched with HCl (50 mL, 2 M) until the reaction became clear and extracted with DCM (3×15 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give ST-200-N19-K7R (27 mg, 14%) as a solid.

[1724] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.36 (m, 1H), 3.87-3.71 (m, 1H), 2.27-2.18 (m, 1H), 2.08-1.88 (m, 4H), 1.87-1.73 (m, 3H), 1.67-1.58 (m, 1H), 1.53-1.34 (m, 9H), 1.31-1.22 (m, 5H), 1.21-1.11 (m, 7H), 1.10-0.98 (m, 3H), 0.95-0.89 (m, 3H), 0.88-0.83 (m, 4H), 0.82-0.74 (m, 1H), 0.67 (s, 3H).

[1725] LCMS Rt=1.246 min in 2.0 min chromatography, 30-90 AB_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.45O [M+H−H.sub.2O].sup.+ 385, found 385.

Example 98: Synthesis of 9810 and 9813

[1726] ##STR00530##

[1727] The synthesis of ST-200-N19-4_7 can be found in Example 94. The synthesis of 6,6-difluoro-1-oxaspiro[2.5]octane can be found in Example 87.

Synthesis of ST-200-N19-K14_1

[1728] ##STR00531##

[1729] n-BuLi (0.656 mL, 2.5 M in hexane, 1.64 mmol) was added to a solution of diisopropylamine (179 mg, 0.442 mmol) in THF (1 mL) under N.sub.2 at −70° C. The mixture was warmed to 25° C. After re-cooling to −70° C., a suspension of ST-200-N19-4_7 (200 mg, 0.412 mmol) in THF (5 mL) was added dropwise under N.sub.2. After stirring at −70° C. for 30 min, 6,6-difluoro-1-oxaspiro[2.5]octane (91.5 mg, 0.618 mmol) was added at −70° C. The reaction mixture was warmed to 25° C. slowly and stirred at 25° C. for 16 hours. The reaction mixture was quenched with saturated NH.sub.4Cl aqueous (15 mL). The mixture was extracted with EtOAc (2×15 mL). The combined organic phase was washed with brine (2×20 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-N19-K14_1 (200 mg, crude) as an oil, which was used directly for the next step.

Synthesis of ST-200-N19-K14

[1730] ##STR00532##

[1731] A solution of ST-200-N19-K14_1 (200 mg, 0.316 umol) in MeOH (30 mL) was heated at 65° C. Mg powder (302 mg, 12.6 mmol) and NiCl.sub.2 (12.1 mg, 0.0948 umol) were added in one portion at 65° C. The mixture was reflux at 65° C. for 1 h and quenched with HCl (50 mL, 2N) until the reaction became clear. The mixture was extracted with DCM (3×20 mL). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by silica gel chromatography (0-15% of EtOAc in PE) to give ST-200-N19-K14 (100 mg, 64%) as a solid.

[1732] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.41-5.36 (m, 1H), 2.27-2.20 (m, 1H), 2.19-2.03 (m, 3H), 2.02-1.88 (m, 5H), 1.86-1.73 (m, 3H), 1.71-1.59 (m, 5H), 1.54-1.33 (m, 10H), 1.32-1.15 (m, 6H), 1.14-1.00 (m, 4H), 0.99-0.97 (m, 1H), 0.96-0.89 (m, 3H), 0.88-0.74 (m, 5H), 0.68 (s, 3H).

[1733] LCMS Rt=1.322 min in 2.0 min chromatography, 30-90 AB_E, purity 100%, MS ESI calcd. for C.sub.31H.sub.47F.sub.2 [M+H−2H.sub.2O].sup.+ 457, found 457.

Synthesis of ST-200-N19-K14A

[1734] ##STR00533##

[1735] Pd(OH).sub.2 (70 mg, dry) was added to a solution of ST-200-N19-K14 (70 mg, 0.142 mmol) in MeOH (20 mL). The mixture was hydrogenated at 50° C. (50 Psi) for 48 hrs. The mixture was filtered, concentrated and purified by combi-flash (0-10% of EtOAc in PE) to give ST-200-N19-K14A (20 mg, 28%) as a solid.

[1736] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.24-1.99 (m, 2H), 1.97-1.73 (m, 6H), 1.71-1.57 (m, 8H), 1.55-1.32 (m, 8H), 1.30-1.16 (m, 4H), 1.13-0.97 (m, 11H), 0.94-0.78 (m, 8H), 0.74-0.55 (m, 5H).

[1737] LCMS Rt=1.344 min in 2.0 min chromatography, 30-90 AB_E, purity 100%, MS ESI calcd. for C.sub.31H.sub.49F.sub.2 [M+H−2H.sub.2O].sup.+ 459, found 459.

Example 99: Synthesis of 9911

[1738] ##STR00534##

[1739] The experimental of intermediate ST-200-N19-4_7 can be found in Example 94.

Synthesis of ST-200-N19-K7S_1

[1740] ##STR00535##

[1741] n-BuLi (0.408 mL, 2.5 M, 1.02 mmol) was added to THF (0.5 mL) under N.sub.2 at −70° C. After that, a suspension of ST-200-N19-47 (200 mg, 0.412 mmol, 1.0 eq.) in THF (1.5 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, (S)-2-methyloxirane (35.8 mg, 0.618 mmol) was added dropwise. Then reaction was stirred at 25° C. for 12 hrs. The reaction was quenched with sat. NH.sub.4Cl (30 mL), extracted with EtOAc (3×15 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude product (250 mg) as a solid, which was used directly for the next step.

Synthesis of ST-200-N19-K7S

[1742] ##STR00536##

[1743] Mg powder (357 mg, 14.7 mmol) was added to a solution of ST-200-N19-K7S_1 (200 mg, 0.368 mmol) and nickel (II) chloride (9.53 mg, 0.074 mmol) in dry methanol (50 mL) under N.sub.2 and the resulting mixture was stirred at 50° C. to initiate continuous hydrogen generation. The reaction mixture was stirred at 60° C. for 1 hour. The reaction mixture was quenched by 2M HCl (100 mL) which was added dropwise at 10° C. until solid was dissolved. After extracting with EtOAc (2×150 mL), the combined organic layer was washed with sat. NaHCO.sub.3 aq. (300 mL), brine (300 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a solid, which was purified by silica gel chromatography (PE/THF=4/1) to give a crude residue, which was purified by re-crystallization from MeCN (10 mL) to afford ST-200-N19-K7S (52 mg, 35%) as a solid.

[1744] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 3.80-3.70 (m, 1H), 2.25-2.20 (m, 1H), 2.05-1.75 (m, 7H), 1.69-1.15 (m, 20H), 1.14-0.70 (m, 13H), 0.67 (s, 3H).

[1745] LCMS Rt=1.241 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.27H.sub.45O [M+H−H.sub.2O].sup.− 385, found 385.

Example 100: Synthesis for 10012, 10042, and 10043

[1746] ##STR00537##

[1747] The experimental of intermediate ST-200-N19-4_7 can be found in Example 94.

Synthesis of ST-200-N19-K9_1

[1748] ##STR00538##

[1749] n-BuLi (0.824 mL, 2.5 M, 2.06 mmol) was added to THF (0.5 mL) under N.sub.2 at −70° C. Next, a suspension of ST-200-N19-4_7 (400 mg, 0.825 mmol) in THF (3.5 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, 2-(tert-butyl) oxirane (123 mg, 1.23 mmol) was added dropwise. Then reaction was stirred at 25° C. for 12 hrs and quenched with sat. NH.sub.4C1 (30 mL), extracted with EtOAc (3×15 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude product (500 mg) as a solid, which was used directly for the next step.

Synthesis of ST-200-N19-K9 (10012)

[1750] ##STR00539##

[1751] Mg powder (663 mg, 27.3 mmol) was added to a solution of ST-200-N19-K9_1 (400 mg, 0.683 mmol) and nickel (II) chloride (17.6 mg, 0.136 mmol) in dry methanol (50 mL) under N.sub.2 and the resulting mixture was stirred at 50° C. to initiate continuous hydrogen generation. The reaction mixture was stirred at 60° C. for 1 hour. The reaction mixture was quenched by 2M HCl (100 mL) which was added dropwise at 10° C. until solid was dissolved. After extracting with EtOAc (2×150 mL), the combined organic layer was washed with sat. NaHCO.sub.3 aq. (300 mL), brine (300 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a solid, which was purified by silica gel chromatography (PE/THF=4/1) to give a crude product, which was re-crystallized from MeCN (10 mL) to afford ST-200-N19-K9 (160 mg, 53%) as a solid.

[1752] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 3.25-3.15 (m, 1H), 2.25-2.20 (m, 1H), 2.10-1.60 (m, 9H), 1.50-1.00 (m, 19H), 0.90-0.75 (m, 18H), 0.67 (s, 3H).

[1753] LCMS Rt=1.412 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.30H.sub.51O [M+H−H.sub.2O].sup.+ 427, found 427.

Synthesis of 10042 and 10043

[1754] ##STR00540##

[1755] ST-200-N19-K9 (120 mg, 0.269 mmol) was purified by SFC (AD (250 mm*30 mm, 10 um), gradient: 40-40% B (A=0.1% NH3/H2O IPA, B=MeOH), flow rate: 60 mL/min) to give ST-200-N19-K9R (Peak 1, 44 mg, 37%) and ST-200-N19-K9S (Peak 2, 45 mg, 38%) as a solid. The stereochemistry at C25 of 10042 and 10043 was confirmed by asymmetric synthesis of 10042 and 10043 from chiral epoxides.

[1756] ST-200-N19-K9R (10042):

[1757] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 3.25-3.15 (m, 1H), 2.25-2.20 (m, 1H), 2.10-1.55 (m, 9H), 1.50-1.30 (m, 8H), 1.29-1.00 (m, 11H), 0.95-0.75 (m, 18H), 0.67 (s, 3H).

[1758] LCMS Rt=1.402 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. for C.sub.30H.sub.51O [M+H−H.sub.2O].sup.+ 427, found 427.

[1759] SFC Rt=6.347 min in 10 min chromatography, AD_3_IPA_DEA_5_40_25ML, 100% de.

[1760] ST-200-N19-K9S (10043):

[1761] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.35 (m, 1H), 3.25-3.15 (m, 1H), 2.25-2.20 (m, 1H), 2.10-1.75 (m, 7H), 1.74-1.35 (m, 7H), 1.34-1.00 (m, 13H), 0.99-0.75 (m, 19H), 0.67 (s, 3H).

[1762] LCMS Rt=1.402 min in 2.0 min chromatography, 30-90 AB, purity 99.2%, MS ESI calcd. for C.sub.30H.sub.51O [M+H−H.sub.2O].sup.− 427, found 427.

[1763] SFC Rt=6.940 min in 10 min chromatography, AD_3_IPA_DEA_5_40_25ML, 99.5% de.

Synthesis for Confirming Stereochemistry

[1764] ##STR00541##

[1765] ST-200-009-001_1 (300 mg, 0.618 mmol) in THF (2 mL, dry) was added to a solution of n-BuLi (0.616 mL, 2.5 M in hexane) in THF (0.5 mL, dry) at −70° C. After stirring at −70° C. for 0.5 h, a solution of (S)-2-(tert-butyl)oxirane (92.7 mg, 0.926 mmol) in THF (0.5 mL, dry) was added dropwise. After stirring at 25° C. for another 16 hrs, the mixture was quenched with sat. NH.sub.4Cl (5 mL). The mixture was extracted with EtOAc (2×10 mL). The combined organic layer was washed with brine (5 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated to give ST-200-089-001_2 (360 mg, crude) as a solid, which was used directly for next step.

[1766] NiCl.sub.2 (8.85 mg, 0.683 mmol) was added to a mixture of ST-200-089-001_2 (360 mg, crude) in MeOH (20 mL) at 25° C. Then the mixture was stirred at 60° C. and Mg powder (828 mg, 34.1 mmol) was added in three bathes for 2 hrs. The reaction was quenched with HCl (1M, 10 mL), the mixture was extracted with EtOAc (2×15 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 flash-combi (0-30% of EtOAc in PE) to give ST-200-089-001 (115 mg, impure) as a solid, which was further purified by SFC (AD (250 mm*30 mm, 10 um), gradient: 40-40% B (A=0.1% NH3/H2O IPA, B=MeOH), flow rate: 50 mL/min) to give ST-200-089-001 (77 mg, 28% yield for 2 steps) as a solid.

[1767] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.41-5.35 (m, 1H), 3.25-3.15 (m, 1H), 2.25-2.20 (m, 1H), 2.10-1.90 (m, 4H), 1.90-1.75 (m, 3H), 1.69-1.50 (m, 3H), 1.50-1.33 (m, 8H), 1.33-1.13 (m, 7H), 1.13-0.98 (m, 4H), 0.98-0.90 (m, 3H), 0.90-0.78 (m, 14H), 0.67 (s, 3H).

[1768] LCMS Rt=1.467 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.30H.sub.51O [M+H−H.sub.2O].sup.+ 427, found 427.

[1769] SFC Rt=6.135 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 100% de.

Example 101: Synthesis for 10114 and 10115

[1770] ##STR00542##

[1771] The synthesis of ST-200-N19-4_7 can be found in Example 94.

Synthesis of ST-200-N19-K4R_1

[1772] ##STR00543##

[1773] n-BuLi (0.408 mL, 2.5 M, 1.02 mmol, 2.5 eq) was added to THF (0.5 mL) under N.sub.2 at −70° C. Next, a suspension of ST-200-N19-4_7 (200 mg, 0.412 mmol, 1.0 eq.) in THF (1.5 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, (S)-2-isopropyloxirane (53.2 mg, 0.618 mmol, 1.5 eq.) was added dropwise. Then reaction was stirred at 25° C. for 12 hrs. The reaction was quenched with sat. NH.sub.4Cl (30 mL), extracted with EtOAc (3×15 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude product (250 mg) as a solid, which was used directly for the next step.

Synthesis of ST-200-N19-K4R

[1774] ##STR00544##

[1775] NiCl.sub.2 (11.3 mg, 0.0874 mmol) and Mg powder (417 mg, 17.4 mmol) were added in one portion to a solution of ST-200-N19-K4R_1 (250 mg, 0.437 mmol) in MeOH (30 mL) at 65° C. The mixture was stirred at 65° C. for 1 h. The mixture was quenched with HCl (50 mL, 2 N) until the reaction became clear and extracted with DCM (3×20 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give ST-200-N19-K4R (38 mg, 20%) as a solid.

[1776] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.41-5.36 (m, 1H), 3.41-3.31 (m, 1H), 2.27-2.20 (m, 1H), 2.07-1.89 (m, 4H), 1.88-1.75 (m, 3H), 1.70-1.59 (m, 3H), 1.54-1.33 (m, 10H), 1.30-1.16 (m, 6H), 1.15-0.97 (m, 4H), 0.95-0.88 (m, 9H), 0.88-0.82 (m, 4H), 0.82-0.74 (m, 1H), 0.68 (s, 3H).

[1777] LCMS Rt=1.362 min in 2.0 min chromatography, 30-90 AB_E, purity 100%, MS ESI calcd. for C.sub.29H.sub.49O [M+H−H.sub.2O].sup.+ 413, found 413.

Synthesis of ST-200-N19-K4S_1

[1778] ##STR00545##

[1779] n-BuLi (0.408 mL, 2.5 M, 1.02 mmol, 2.5 eq) was added to THF (0.5 mL) under N.sub.2 at −70° C. Next, a suspension of ST-200-N19-4_7 (200 mg, 0.412 mmol, 1.0 eq.) in THF (1.5 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, (R)-2-isopropyloxirane (53.2 mg, 0.618 mmol, 1.5 eq.) was added dropwise. Then reaction was stirred at 25° C. for 12 hrs. The reaction was quenched with sat. NH.sub.4Cl (30 mL), extracted with EtOAc (3×15 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude product (250 mg) as a solid, which was used directly for the next step.

Synthesis of ST-200-N19-K4S

[1780] ##STR00546##

[1781] NiCl.sub.2 (11.3 mg, 0.0874 mmol) and Mg powder (417 mg, 17.4 mmol) were added in one portion to a solution of ST-200-N19-K4S_1 (250 mg, 0.437 mmol) in MeOH (30 mL) at 65° C. The mixture was stirred at 65° C. for 1 h. The mixture was quenched with HCl (50 mL, 2 N) until the reaction became clear and extracted with DCM (3×20 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give ST-200-N19-K4S (36 mg, 19%) as a solid.

[1782] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.41-5.36 (m, 1H), 3.41-3.31 (m, 1H), 2.27-2.20 (m, 1H), 2.07-1.89 (m, 4H), 1.88-1.75 (m, 3H), 1.71-1.57 (m, 3H), 1.55-1.35 (m, 9H), 1.34-1.23 (m, 4H), 1.22-0.99 (m, 7H), 0.95-0.89 (m, 9H), 0.88-0.82 (m, 4H), 0.82-0.74 (m, 1H), 0.68 (s, 3H).

[1783] LCMS Rt=1.356 min in 2.0 min chromatography, 30-90 AB_E, purity 100%, MS ESI calcd. for C.sub.29H.sub.49O [M+H−H.sub.2O].sup.+ 413, found 413.

Example 102: Synthesis for 10216

[1784] ##STR00547##

[1785] The synthesis of ST-200-N19-4_7 can be found in Example 94.

Synthesis of ST-200-N19-K15_1

[1786] ##STR00548##

[1787] n-BuLi (0.408 mL, 2.5 M, 1.02 mmol, 2.5 eq) was added to THF (0.5 mL) under N.sub.2 at −70° C. Next, a suspension of ST-200-N19-4_7 (200 mg, 0.412 mmol, 1.0 eq.) in THF (1.5 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, 1,6-dioxaspiro[2.5]octane (70.5 mg, 0.618 mmol, 1.5 eq.) was added dropwise. Then reaction was stirred at 25° C. for 12 hrs. The reaction was quenched with sat. NH.sub.4Cl (30 mL), extracted with EtOAc (3×15 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude product (250 mg) as a solid, which was used directly for the next step.

[1788] C3

Synthesis of ST-200-N19-K15

[1789] ##STR00549##

[1790] NiCl.sub.2 (10.8 mg, 0.0834 mmol) and Mg powder (398 mg, 16.6 mmol) were added in one portion to a solution of ST-200-N19-K15_1 (250 mg, 0.417 mmol) in MeOH (30 mL) at 65° C. The mixture was stirred at 65° C. for 1 h. The mixture was quenched with HCl (50 mL, 2 N) until the reaction became clear and extracted with DCM (3×20 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give ST-200-N19-K15 (43 mg, 22%) as a solid.

[1791] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.44-5.33 (m, 1H), 3.81-3.69 (m, 4H), 2.27-2.19 (m, 1H), 2.07-1.88 (m, 4H), 1.87-1.76 (m, 3H), 1.71-1.58 (m, 3H), 1.53-1.32 (m, 11H), 1.31-1.13 (m, 7H), 1.12-1.07 (m, 2H), 1.06-0.95 (m, 3H), 0.94-0.89 (m, 3H), 0.88-0.82 (m, 4H), 0.82-0.75 (m, 1H), 0.68 (s, 3H).

[1792] LCMS Rt=1.200 min in 2.0 min chromatography, 30-90 AB_E, purity 100%, MS ESI calcd. for C.sub.30H.sub.47O [M+H−2H.sub.2O].sup.+ 423, found 423.

Example 103: Synthesis for 10317

[1793] ##STR00550##

[1794] The synthesis of ST-200-N19-4_7 can be found in Example 94. The synthesis of the epoxide can be found in Example 28.

Synthesis of ST-200-N19-K17_1

[1795] ##STR00551##

[1796] n-BuLi (0.408 mL, 2.5 M, 1.02 mmol, 2.5 eq) was added to THF (0.5 mL) under N.sub.2 at −70° C. Next, a suspension of ST-200-N19-47 (200 mg, 0.412 mmol, 1.0 eq.) in THF (1.5 mL) was added dropwise to give a suspension. After stirring at −70° C. for 30 min, 6,6-dimethyl-1-oxaspiro[2.5]octane (86.6 mg, 0.618 mmol, 1.5 eq.) was added dropwise. Then reaction was stirred at 25° C. for 12 hrs. The reaction was quenched with sat. NH.sub.4Cl (30 mL), extracted with EtOAc (3×15 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude product (250 mg) as a solid, which was used directly for the next step.

Synthesis of ST-200-N19-K17

[1797] ##STR00552##

[1798] NiCl.sub.2 (10.3 mg, 0.08 mmol) and Mg powder (384 mg, 16.0 mmol) were added in one portion to a solution of ST-200-N19-K17_1 (250 mg, 0.4 mmol) in MeOH (30 mL) at 65° C. The mixture was stirred at 65° C. for 1 h. The mixture was quenched with HCl (50 mL, 2 N) until the reaction became clear and extracted with DCM (3×20 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give impure ST-200-N19-K17 (58 mg, 30%) as a solid, which was triturated with hexane (3 mL). The mixture was filtered to give pure ST-200-N19-K17 (33 m gas a solid.

[1799] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.44-5.33 (m, 1H), 2.27-2.2 (m, 1H), 2.07-1.88 (m, 4H), 1.87-1.75 (m, 3H), 1.68-1.57 (m, 2H), 1.52-1.45 (m, 7H), 1.44-1.32 (m, 7H), 1.28-1.15 (m, 8H), 1.14-0.95 (m, 6H), 0.94-0.89 (m, 6H), 0.89-0.82 (m, 7H), 0.82-0.74 (m, 1H), 0.68 (s, 3H).

[1800] LCMS Rt=1.516 min in 2.0 min chromatography, 30-90 AB_E, purity 100%, MS ESI calcd. for C.sub.33H.sub.53 [M+H−2H.sub.2O].sup.+ 449, found 449.

Example 104: Synthesis for 10456

[1801] ##STR00553##

[1802] The synthesis of ST-200-N19-4_7 can be found in Example 94.

Synthesis of ST-200-52-8_1

[1803] ##STR00554##

[1804] n-BuLi (0.246 mL, 2.5 M, 0.617 mmol) was added to THF (0.3 mL) under N.sub.2 at −70° C. A solution of ST-200-N19-4_7 (0.12 g, 0.247 mmol) in THF (1 mL) was added at −70° C. After stirring at −70° C. for 1 hour, (R)-2-(trifluoromethyl)oxirane (41.4 mg, 0.37 mmol) was added at −70° C. The reaction mixture was warmed to 15° C. and stirred at 15° C. for 18 hrs. The reaction mixture was quenched with saturated NH.sub.4Cl aqueous (6 mL) at 0° C. The mixture was extracted with EtOAc (2×8 mL). The combined organic phase was washed with brine (2×10 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give a crude product ST-200-52-8_1 (150 mg, crude) as a solid, which was used directly for the next step.

Synthesis of ST-200-52-8

[1805] ##STR00555##

[1806] NiCl.sub.2 (6.5 mg, 0.0502 mmol) and Mg powder (240 mg, 10.0 mmol) were added in one to a solution of ST-200-52-8_1 (150 mg, 0.251 mmol) in MeOH (30 mL) at 65° C. The mixture was stirred at 65° C. for 1 h. The mixture was quenched with HCl (50 mL, 2 N) until the reaction became clear and extracted with DCM (3×20 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give ST-200-52-8 (20 mg, 17%) as a solid.

[1807] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.43-5.34 (m, 1H), 3.98-3.85 (m, 1H), 2.27-2.19 (m, 1H), 2.06-1.88 (m, 5H), 1.87-1.76 (m, 3H), 1.68-1.57 (m, 4H), 1.53-1.34 (m, 8H), 1.29-1.15 (m, 5H), 1.13-0.98 (m, 4H), 0.95-0.89 (m, 3H), 0.89-0.82 (m, 4H), 0.81-0.74 (m, 1H), 0.68 (s, 3H).

[1808] LCMS Rt=1.255 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. for C.sub.27H.sub.42F.sub.3O [M+H−H.sub.2O].sup.+ 439, found 439.

Example 105: Synthesis for 10557

[1809] ##STR00556##

[1810] The synthesis of ST-200-N19-4_7 can be found in Example 94. The synthesis of the tosylate can be found in Example 30.

Synthesis of ST-200-N19-K11_1

[1811] ##STR00557##

[1812] n-BuLi (0.656 mL, 2.5 M in hexane, 1.64 mmol) was added to a solution of diisopropylamine (179 mg, 1.77 mmol) in THF (1 mL) under N.sub.2 at −70° C. The mixture was warmed to 25° C. After re-cooling to −70° C., a suspension of ST-200-N19-47 (200 mg, 0.412 mmol) in THF (5 mL) was added dropwise under N.sub.2 at −70° C. After stirring at −70° C. for 30 min, (S)-3,3,3-trifluoro-2-hydroxy-2-methylpropyl 4-methylbenzenesulfonate (184 mg, 0.618 mmol) was added. The reaction mixture was warmed to 25° C. slowly and stirred at 25° C. for 16 hrs. The reaction mixture was quenched with saturated NH.sub.4Cl aqueous (15 mL). The mixture was extracted with EtOAc (2×15 mL). The combined organic phase was washed with brine (2×20 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-N19-K11_11 (200 mg, crude) as an oil, which was used for the next step directly.

Synthesis of ST-200-N19-K11

[1813] ##STR00558##

[1814] A solution of ST-200-N19-K11_1 (200 mg, 0.327 mmol) in MeOH (30 mL) was heated at 65° C. Mg powder (312 mg, 13.0 mmol) and NiCl.sub.2 (12.5 mg, 0.0981 mmol) were added in one portion at 65° C. The mixture was refluxed at 65° C. for 1 h. The mixture was quenched with HCl (50 mL, 2N) until the reaction became clear and extracted with DCM (3×20 mL). The combined organic layer was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by silica gel chromatography (0-15% of EtOAc in PE) to give impure ST-200-N19-K11 (26 mg, 17%) as a solid, which was triturated with hexane (3 mL) to give ST-200-N19-K11 (13 mg, 50%) as a solid.

[1815] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.41-5.36 (m, 1H), 2.27-2.19 (m, 1H), 2.08-1.95 (m, 3H), 1.93-1.75 (m, 5H), 1.69-1.57 (m, 4H), 1.55-1.37 (m, 7H), 1.34 (s, 3H), 1.30-1.15 (m, 6H), 1.14-0.97 (m, 4H), 0.96-0.89 (m, 3H), 0.88-0.74 (m, 5H), 0.68 (s, 3H).

[1816] LCMS Rt=1.261 min in 2.0 min chromatography, 30-90AB_E, purity 100%, MS ESI calcd. for C.sub.28H.sub.44F.sub.3O [M+H−H.sub.2O].sup.+ 453, found 453.

Example 106: Synthesis for 10673

[1817] ##STR00559##

[1818] The synthesis of 200-N19-4_7 can be found in Example 94.

Synthesis of ST-200-N19-K5_1

[1819] ##STR00560##

[1820] A suspension of ST-200-N19-47 (400 mg, 0.8251 mmol) in THF (4 mL) was added dropwise at −70° C. under N.sub.2 to a solution of n-BuLi (5.0 mL, 2.5 M in hexane, 6.5 mmol) in THF (2 mL) inside sealed tube (10 mL). The mixture was stirred for 30 minutes at −70° C. A solution of diisopropylamine (274 mg, 2.7 mmol) was added dropwise at −70° C., then a solution of 2-ethyloxirane(178 mg, 2.47 mmol) was added dropwise at −70° C. The mixture was stirred for another 30 min and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for hour. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL), extracted with EtOAc (3×50 mL). The combined organic phase was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-N19-K5_1 (450 mg, crude), which was used directly for the next step.

Synthesis of ST-200-N19-K5

[1821] ##STR00561##

[1822] A solution of ST-310-N19-K5_1 (0.45 g, 0.808 mmol) in MeOH (30 mL) was heated at 60° C. Mg powder (775 mg, 32.3 mmol) was added in four portions at 60° C. The mixture was stirred at 60° C. for 1 h. The mixture was quenched with HCl (30 mL, 2 M) until the reaction became clear and extracted with DCM (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-30% of EtOAc in PE) to give 330 mg of impure product, which was purified by flash column (0-20% of EtOAc in PE) to give pure 150 mg of a solid. The solid (150 mg, 0.873 mmol) in THF (5 mL) was added lindlar catalyst (100 mg). The mixture was stirred at 25° C. under H.sub.2 (15 Psi) for 16 hrs. The mixture was filtered and concentrated to give ST-200-N19-K5 (100 mg, crude) as a solid. The residue was triturated from n-hexane (2 mL) at 20° C. to give ST-200-N19-K5 (50 mg) as solid 1H NMR (400 MHz, CDCl.sub.3) δ 5.39-5.37 (m, 1H), 3.51 (s, 1H), 2.25-2.21 (m, 1H), 2.05-1.70 (m, 7H), 1.65-1.50 (m, 4H), 1.50-1.25 (m, 10H), 1.25-1.15 (m, 6H), 1.14-1.10 (m, 4H), 0.94-0.91 (m, 6H), 0.88-0.83 (m, 5H), 0.67 (s, 3H).

[1823] LCMS Rt=1.269 min in 2 min chromatography, 30-90AB_2MIN_E, purity 99%, MS ESI calcd. For C.sub.28H.sub.47O.sup.+ [M+H−H.sub.2O]+ 399, found 399.

Example 107: Synthesis for 10790

[1824] ##STR00562## ##STR00563##

[1825] The experimental of intermediate ST-200-N19-4_3 can be found in Example 94.

Synthesis of ST-200-52-3_1, ST-200-52-3_1A

[1826] ##STR00564##

[1827] 9-BBN dimer (2.22 g, 9.12 mmol) was added to a solution of 200-N19-4_3 (2 g, 6.08 mmol) in anhydrous THF (20 mL) under nitrogen atmosphere at 30° C. The mixture was stirred at 65° C. for 3 hours. The reaction mixture was cooled to 30° C., and (Z)-methyl 3-bromoacrylate (1.20 g, 7.29 mmol), CsF (1.83 g, 12.1 mmol) and Pd(t-Bu.sub.3P).sub.2(310 mg, 0.608 mmol) were added to the mixture. The resulting mixture was stirred at 65° C. for 16 hours. The reaction was cooled, quenched with water (300 mL), extracted with EtOAc (3×300 mL). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give a residue, which was purified by combi-flash (5%-15% of EtOAc in PE) to give ST-200-52-3_1 (400 mg, 16%) as a solid, ST-200-52-3_1A (340 mg, 13%) as solid, and the mixture of ST-200-52-31 and ST-200-52-3_1A (340 mg) as a solid.

[1828] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.29-6.23 (m, 1H), 5.82 (d, J=12.0 Hz, 1H), 5.39-5.34 (m, 1H), 3.70 (s, 3H), 2.65-2.55 (m, 2H), 2.23 (dd, J=3.2, 13.2 Hz, 1H), 2.08-1.78 (m, 8H), 1.72-1.58 (m, 5H), 1.53-1.38 (m, 6H), 1.37-0.99 (m, 5H), 0.95 (d, J=6.4 Hz, 3H), 0.86 (t, J=7.2 Hz, 3H), 0.70 (s, 3H).

Synthesis of ST-200-52-3_2

[1829] ##STR00565##

[1830] ST-200-52-3_1 (340 mg, 0.8200 mmol) in THF/MeOH (8 mL/8 mL) and Lindlar (600 mg) was stirred at 20° C. under H.sub.2 (15 psi) for 16 hours. The reaction mixture was filtered and the filtrate cake was washed with DCM (2×10 mL), MeOH (2×10 mL), THF (2×10 mL). The filtrate was concentrated under vacuum to give ST-200-52-3_2 (300 mg, crude) as an oil, which was triturated in MeOH/H.sub.2O (5 mL/5 mL) to give ST-200-52-3_2 as an oil. The crude oil was further purified by flash column (0-10% of EtOAc in PE) to give ST-200-52-3_2 (240 mg, 80%) as a solid.

[1831] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.44-5.35 (m, 1H), 3.66 (s, 3H), 2.38-2.17 (m, 3H), 2.02-1.76 (m, 7H), 1.61-1.37 (m, 10H), 1.28-0.99 (m, 9H), 0.96-0.76 (m, 8H), 0.74-0.65 (m, 3H).

Synthesis of 10790

[1832] ##STR00566##

[1833] Ti(i-PrO).sub.4 (163 mg, 0.58 mmol) and EtMgBr (0.7 mL, 3 M in Et.sub.2O, 2 mmol) were added to a solution of ST-200-52-3_2 (240 mg, 0.58 mmol) in THF (2 mL) at 25° C. The reaction mixture was stirred at 25° C. for 15 min under N.sub.2. The reaction mixture was quenched with saturated aqueous NH.sub.4Cl (10 mL) solution and extracted with EtOAc (3×20 mL). The combined organic layer was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give a crude product, which was purified by flash column (0-15% of EtOAc in PE) to afford a crude a solid. The solid was further triturated from MeCN (5 mL) and purified by silica gel chromatography (PE/EtOAc=10/1) to afford ST-200-52-3 (15 mg, 6%) as a solid.

[1834] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.44-5.35 (m, 1H), 2.27-2.19 (m, 1H), 2.07-1.61 (m, 10H), 1.51-1.34 (m, 8H), 1.31-0.98 (m, 11H), 0.97-0.92 (m, 3H), 0.89-0.82 (m, 5H), 0.76-0.71 (m, 2H), 0.68 (s, 3H), 0.48-0.39 (m, 2H).

[1835] LCMS Rt=1.226 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.28H.sub.45O [M+H−H.sub.2O].sup.+ 397, found 397.

Example 108: Synthesis of 10841

[1836] ##STR00567##

[1837] The experimental of intermediate ST-200-N19-4_7 can be found in Example 94.

Synthesis of ST-200-091-002_2

[1838] ##STR00568##

[1839] To a solution of ST-200-N19-4_7 (500 mg, 1.03 mmol) in anhydrous THF (3 mL) was added n-BuLi (1.23 mL, 3.09 mmol, 2.5M in n-hexane) drop-wise at −70° C. under N.sub.2. The reaction mixture was stirred at −70° C. for 30 mins. To the mixture was added a solution of (R)-2-ethyloxirane (111 mg, 1.54 mmol) in anhydrous THF (0.5 mL) drop-wise at −70° C. and stirred for another 1 h. The reaction mixture was then stirred at 25° C. for 12 h. The reaction was quenched with saturated aqueous NH.sub.4Cl (50 mL). The aqueous phase was extracted with EtOAc (3×50 mL). The combine organic phase was washed whit saturated brine (2×50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give ST-200-091-002_2 (0.5 g, crude) as an oil.

Synthesis of ST-200-091-002

[1840] ##STR00569##

[1841] To a solution of ST-200-091-002_2 (0.5 g, crude) in MeOH (50 mL) was added Mg powder (1.07 g, 44.8 mmol) and NiCl.sub.2 (20 mg) at 25° C. under N.sub.2. After stirring at 50° C. for 1 h, the reaction mixture was quenched with HCl (100 mL, 1 M) until the reaction became clear. The aqueous phase was extracted with EtOAc (3×80 mL). The combined organic phase was washed with saturated brine (2×50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=10/1 to 8/1) to afford ST-200-091-002 (120 mg, 32%) as a solid, which was purified by SFC (Column: AD (250 mm*30 mm, 5 um), Condition: 0.1% NH.sub.3H.sub.2O ETOH, Begin B: 40%, End B: 40%) to afford ST-200-091-002 (50 mg, 42.0%) as a solid.

[1842] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.38-5.36 (m, 1H), 3.55-3.49 (m, 1H), 2.29-2.20 (m, 1H), 2.05-1.75 (m, 7H), 1.65-1.31 (m, 13H), 1.30-1.15 (m, 7H), 1.14-0.76 (m, 15H), 0.68 (s, 3H).

[1843] LCMS Rt=1.326 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. for C.sub.28H.sub.47O [M+H−H.sub.2O].sup.+ 399, found 399.

Example 109: Synthesis of 10949

[1844] ##STR00570##

[1845] The experimental of intermediate ST-200-N19-4_7 can be found in Example 94.

Synthesis of ST-200-091-001_2

[1846] ##STR00571##

[1847] To a THF (0.5 mL) under N.sub.2 at −70° C. was added n-BuLi (2.5 M, 2.57 mmol, 1.02 mL). After that, a suspension of ST-200-N19-4_7 (500 mg, 1.03 mmol) in THF (3 mL) was added drop-wise to give a suspension. After stirring at −70° C. for 30 min, a solution of (S)-2-ethyloxirane (88.6 mg, 1.23 mmol) in THF (0.5 mL) was added. Then reaction was stirred at stirred at 25° C. for 16 hours. The mixture was poured into ice-water (20 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to afford ST-200-091-001_2 (580 mg, crude) as a solid, which was used directly for the next step.

Synthesis of ST-200-091-001

[1848] ##STR00572##

[1849] To a solution of ST-200-091-001_2 (580 mg, 4.41 mmol) and nickel (II) chloride (6.73 mg, 0.052 mmol) in MeOH (30 mL) was added Mg powder (499 mg, 20.8 mmol) at 25° C. The mixture was stirred at 50° C. for 1 h. After cooling, the mixture was quenched with HCl (100 mL, 2M) until the reaction became clear and extracted with EtOAc (2×50 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by a silica gel column (PE/EtOAc=10/1 to 3/1) to give ST-200-091-001 (120 mg, 28%) as a solid.

[1850] The ST-200-091-001 (120 mg, 0.287 mmol) was separated by SFC (column: AD (250 mm*30 mm, 5 um)), gradient: 50-50% B (A=0.1% NH.sub.3H.sub.2O ETOH), flow rate: 60 mL/min) to give ST-200-091-001 (58 mg, 14%) as a solid.

[1851] .sup.1HNMR (400 MHz, CDCl3) δ 5.42-5.38 (m, 1H), 3.57-3.47 (m, 1H), 2.25-2.21 (m, 1H), 2.07-1.88 (m, 4H), 1.86-1.78 (m, 3H), 1.68-1.59 (m, 2H), 1.52-1.31 (m, 12H), 1.29-0.98 (m, 10H), 0.96-0.75 (m, 11H), 0.68 (s, 3H).

[1852] LCMS Rt=1.318 min in 2.0 min chromatography, 30-90 AB, purity 100%, MS ESI calcd. For C.sub.28H.sub.47O [M−H.sub.2O+H]+ 399, found 399.

[1853] SFC Rt=6.962 min in 10 min chromatography, AD_3_EtOH_DEA_5_40_25ML, 99.4% de

Example 110. Biological Data

[1854] Experiments conducted as described in Example 2 and results are reported in Table 2-62.

TABLE-US-00006 TABLE 2-62 Avg EC50 Avg Avg Avg 2A Emax EC50 Emax Compound (nM) 2A (%) 2B (nM) 2B (%) [00573] 194 200.3 161.8 143.5 192.5 [00574] 9567 327.5 104.0 378.1 124.3 [00575] 9670 366.1 153.8 219.9 213.2 [00576] 9792 334.2 64.5 284.9 58.2 [00577] 9810 124.0 87.1 151.2 91.6 [00578] 9911 252.6 233.7 287.6 326 [00579] 10012 790.0 154.1 909.6 167.6 [00580] 9813 469.2 68.7 267.7 99.9 [00581] 10114 320.8 146.3 505.6 179.1 [00582] 10115 555.9 191.6 458.2 225.6 [00583] 10216 1196.8 79.3 445.6 120.7 [00584] 10317 >10000 56.7 >10000 59.9 [00585] 10042 >10000 65.9 1877.4 112.8 [00586] 10043 897.3 145.7 1067.0 210.6 [00587] 10456 239.6 230.6 208.6 265.7 [00588] 10557 286.8 189.7 191.7 202.5 [00589] 10673 253.6 215.5 180.1 213.5 [00590] 10790 203.8 159.8 270.5 245.8 [00591] 10841 >10000 15.6 >10000 30.1 [00592] 10949 66.4 97.4 138.1 147.4

Example 111. Synthesis of Compound A-1

[1855] ##STR00593##

Step 1

[1856] To a solution of S,S-cat (2 g, 3.65 mmol) in anhydrous DCM (30 mL) was added a solution of cobalt(II) acetate (775 mg, 4.38 mmol) in MeOH (30 mL) under nitrogen at 20° C. The mixture was stirred for 30 mins at 20° C. and at 0° C. for 1 h. The precipitated solid was filtered, washed with cold MeOH (2×30 mL) and dried in vacuum to give Co-S,S-cat (1.6 g, 73%) as a solid.

Step 2

[1857] To a solution of Co-S,S-cat (1.07 g, 1.78 mmol) in toluene (30 mL) was added AcOH (1.12 g, 18.7 mmol). The mixture was stirred at 20° C. for 30 mins. The solution was concentrated in vacuum to give a crude solid. The resulting catalyst residue was dissolved in neat A-0 (100 g, 892 mmol) at 20° C., the reaction mixture was cooled to 0° C., and water (8.82 g, 490 mmol) was added dropwise. The mixture was warmed to 20° C. and stirred for 48 hrs. A-1 (44 g) was isolated by distillation from the reaction mixture.

[1858] .sup.1H NMR (400 MHz, DMSO-d6) δ 3.96 (s, 1H), 3.11-2.98 (m, 2H).

[1859] The e.e. of A-1 was determined by opening the epoxide with benzylamine. A-1 (200 mg, 1.78 mmol) was added to dry benzylamine (190 mg, 1.78 mmol), and the mixture was stirred at 20° C. for 2 hrs. A solid precipitated, which was triturated from petroleum ether to afford the product (260 mg, 67%) as a solid. The e.e. of this product was determined to be 100% by chiral HPLC. (Column: CD-PH 250*4.6 mm I.D., 5 um; Mobile phase: from 10% to 80% of B in A (A: Water with 0.069% TFA B: Acetonitrile); Flow rate: 0.8 mL/min; Column Temperature: 30° C.).

Example 112. Synthesis of Compound 112

[1860] ##STR00594## ##STR00595## ##STR00596##

Step 1

[1861] To a mixture of MePPh.sub.3Br (1.28 kg, 3.6 mol) in THF (4.5 L) was added t-BuOK (404 g, 3.6 mol) at 15° C. under N.sub.2. The resulting mixture was stirred at 50° C. for 30 min. Pregnenolone (950 g, 2.9 mol) was added in portions below 65° C. The reaction mixture was stirred at 50° C. for 1 hour. The combined mixture was quenched with saturated NH.sub.4Cl aqueous (1 L) at 15° C. THF layer was separated. The aqueous was extracted with EtOAc (2×2 L). The combined organic phase was concentrated under vacuum to give a solid. The solid was further purified by trituration with MeOH/H.sub.2O (1:1, 15 L) at reflux to give B-1C (940 g, 99%) as a solid.

[1862] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.32 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H), 3.58-3.46 (m, 1H), 2.36-2.16 (m, 2H), 2.08-1.94 (m, 2H), 1.92-1.62 (m, 9H), 1.61-1.39 (m, 6H), 1.29-1.03 (m, 4H), 1.01 (s, 3H), 0.99-0.91 (m, 1H), 0.59 (s, 3H).

Step 2

[1863] To a solution of B-1C (4 kg, 12.7 mol) in DCM (30 L) was added imidazole (1.72 kg, 25.4 mol) and TBSCl (2.86 kg, 19.0 mol) at 25° C. The reaction mixture was stirred at 25° C. for 16 hrs. The reaction mixture was treated with water (10 L). The organic phase was concentrated to give crude product which was triturated in MeOH (15 L) at reflux to give B-2C (5.02 kg, 92%) as a solid.

[1864] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.38-5.28 (m, 1H), 4.85 (s, 1H), 4.71 (s, 1H), 3.57-3.41 (m, 1H), 2.33-2.11 (m, 2H), 2.10-1.94 (m, 2H), 1.90-1.61 (m, 8H), 1.60-1.38 (m, 6H), 1.28-1.03 (m, 4H), 1.00 (s, 3H), 0.98-0.91 (m, 1H), 0.89 (s, 9H), 0.58 (s, 3H), 0.06 (s, 6H).

Step 3

[1865] To a solution of B-2C (1.69 kg, 3.94 mol) in THF (8 L) was added 9-BBN dimer (671 g, 2.75 mol) and the reaction was stirred at 25° C. under N.sub.2 for 1 h and a solid was formed. To the reaction mixture was added ethanol (2.26 L, 39.4 mol) and NaOH (3.94 L, 5 M, 19.7 mol) and the mixture became clear. Then H.sub.2O.sub.2 (3.94 L, 10 M, 39.4 mol) was added dropwise at 25° C. and the inner temperature was raised to reflux. The mixture was cooled and stirred for 16 hrs and a solid was formed. This was followed by adding Na.sub.2SO.sub.3 (2.5 L, 20% aq.) and water (5 L) at 25° C. The mixture was stirred for 1 h. After the stirrer was turned off, a clear lower layer and another upper suspension layer were formed. The clear lower layer was discarded. The upper suspension layer was treated with water (20 L). The mixture was stirred for 15 mins. The mixture was filtered. The solid was washed with water until pH<9 to give the wet product. The wet product B-3C was dissolved in DCM (100 L). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered and concentrated to 20 L. The residue was used in the next step directly.

[1866] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.40-5.23 (m, 1H), 3.70-3.60 (m, 1H), 3.55-3.42 (m, 1H), 3.41-3.31 (m, 1H), 2.31-2.20 (m, 1H), 2.20-2.11 (m, 1H), 2.06-1.91 (m, 2H), 1.89-1.67 (m, 3H), 1.65-1.39 (m, 7H), 1.38-1.08 (m, 6H), 1.05 (d, J=6.4 Hz, 3H), 1.00 (s, 3H), 0.99-0.91 (m, 2H), 0.88 (s, 9H), 0.70 (s, 3H), 0.05 (s, 6H).

Step 4

[1867] To a solution of B-3C (theoretical mass: 5.2 kg, 11.6 mol) in DCM (15 L) was added N-methyl-imidazole (1.37 L, 17.4 mol) and TEA (3.2 L, 23.2 mol) at 25° C. Then TsCl (2.53 kg, 13.3 mol) was added into the solution in portions to keep the inner temperature between 25 to 30° C. The reaction mixture was stirred at 25° C. for 1 h. To the mixture was added water (10 L), citric acid (20%, 1 L). HCl (1 M) was added till pH=3. The organic layer was separated, washed with water (2×10 L), NaHCO.sub.3 (sat. aq. 5 L) and brine (5 L), dried over Na.sub.2SO.sub.4, filtered and concentrated to give B-4C (6.63 kg, 95% for 2 steps) as a solid.

[1868] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.78 (d, J=8.4 Hz, 2H), 7.34 (d, J=8.0 Hz, 2H), 5.37-5.25 (m, 1H), 3.96 (dd, J=2.8, 9.2 Hz, 1H), 3.79 (dd, J=6.4, 9.2 Hz, 1H), 3.53-3.41 (m, 1H), 2.45 (s, 3H), 2.32-2.20 (m, 1H), 2.20-2.11 (m, 1H), 2.01-1.88 (m, 2H), 1.84-1.61 (m, 4H), 1.56-1.31 (m, 6H), 1.23-1.02 (m, 5H), 1.02-0.95 (m, 7H), 0.93-0.90 (m, 1H), 0.88 (s, 9H), 0.63 (s, 3H), 0.05 (s, 6H).

Step 5

[1869] To a solution of B-4C (10 g, 16.6 mmol) in DMF (300 mL) was added KI (6.88 g, 41.5 mmol). The suspension was stirred at 50° C. for 2 hrs. The mixture was quenched with water (500 mL) and extracted with PE (3×150 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was triturated with EtOAc (50 mL) and filtered to give B-5C (7 g, 76%) as a solid.

[1870] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.32-5.28 (m, 1H), 3.52-3.40 (m, 1H), 3.38-3.30 (m, 1H), 3.20-3.11 (m, 1H), 2.32-2.10 (m, 2H), 2.00-1.90 (m, 1H), 1.90-1.35 (m, 13H), 1.30-1.15 (m, 4H), 1.15-0.96 (m, 6H), 0.96-0.80 (m, 10H), 0.71 (s, 3H), 0.06 (s, 6H).

Step 6

[1871] To a solution of (methylsulfonyl) benzene (24.5 g, 157 mmol) in THF (200 mL) was added n-BuLi (57.2 mL, 143 mmol) at −70° C. under N.sub.2. The mixture was stirred at −70° C. for 30 minutes. A solution of B-5C (40 g, 71.8 mmol) in THF (200 mL) was added dropwise at 25° C. After addition, the reaction was allowed to stir at 25° C. for 3h. The reaction was quenched with sat. NH.sub.4Cl (50 mL) and extracted with EtOAc (2×20 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated to give B-6C (40 g, 95%).

[1872] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.90-7.86 (m, 2H), 7.68-7.61 (m, 1H), 7.60-7.52 (m, 2H), 5.33-5.29 (m, 1H), 3.50-3.42 (m, 1H), 3.17-3.06 (m, 1H), 3.04-2.96 (m, 1H), 2.31-2.11 (m, 3H), 1.99-1.90 (m, 2H), 1.87-1.67 (m, 5H), 1.51-1.40 (m, 7H); 1.24-0.82 (m, 27H); 0.68-0.58 (m, 3H).

Step 7

[1873] To a suspension of B-6C (31.0 g, 54.7 mmol) in THF (100 mL) was added TBAF (21.4 kg, 82.0 mmol). The mixture was stirred at 65° C. for 1 h. The mixture turned clear. To the mixture was added water (300 mL) and stirred at 80° C. for 2 h. The mixture was filtered after cooling. The solid was washed with water (300 ml), dried in air to give B-7C (17 g, crude) as a solid, which was used directly for the next step.

Step 8

[1874] To a solution of B-7C (10.5 g, 22.3 mmol) in MeOH (100 mL) and THF (100 mL) was added Pd/C (2 g, <1% water). Then the solution was hydrogenated under 30 psi of hydrogen at 25° C. for 48 h. The reaction was conducted in parallel twice. The mixture was filtered through a pad of celite and the filtrate was concentrated in vacuum. The product was triturated by MeCN (100 mL) to afford B-8C (15 g, 71%) as a solid.

[1875] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.91-7.89 (m, 2H), 7.67-7.64 (m, 1H), 7.59-7.55 (m, 2H), 3.60-3.55 (m, 1H), 3.15-3.06 (m, 1H), 3.02-2.94 (m, 1H), 1.89-1.70 (m, 3H), 1.69-1.64 (m, 3H), 1.53-1.20 (m, 12H), 1.15-0.89 (m, 7H), 0.86 (m, 4H), 0.78 (s, 3H), 0.63-0.56 (m, 4H)

Step 9

[1876] To a solution of B-8C (5.0 g, 10.5 mmol) in DCM (50 mL) was added DMP (8.90 g, 21.0 mmol) at 25° C. The mixture was stirred at 25° C. for 30 mins. The reaction was quenched with saturated aqueous NaHCO.sub.3 (50 mL) and the pH was adjusted to 7˜8. Then saturated Na.sub.2S.sub.2O.sub.3 (100 mL) was added to the solution and extracted with DCM (100 mL). The combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give B-9C (4.6 g, crude) as a solid, which was used in the next step without further purification.

[1877] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.96-7.86 (m, 2H), 7.61-7.53 (m, 3H), 3.17-3.04 (m, 1H), 3.03-2.94 (m, 1H), 2.42-2.15 (m, 3H), 2.10-1.80 (m, 4H), 1.73-1.63 (m, 2H), 1.60 (s, 3H), 1.55-1.45 (m, 4H), 1.40-1.24 (m, 5H), 1.24-1.15 (m, 1H); 1.14-1.03 (m, 2H); 0.99 (s, 3H); 0.90-0.84 (m, 4H); 0.74-0.65 (m, 1H); 0.63 (s, 3H).

Step 10

[1878] To a solution of B-9C (4.6 g, 9.77 mmol) and CsF (2.96 g, 19.5 mmol) in THF (50 mL) was added dropwise TMSCF.sub.3 (2.77 g, 19.5 mmol) at 0° C. The mixture was stirred and kept below 10° C. for 30 mins. To the mixture was added TBAF (24.4 mL, 1 M in THF, 24.4 mmol) at 10° C. The mixture was stirred and kept below 10° C. for 30 mins. To the mixture was added water (20 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with brine (50 mL) and dried over Na.sub.2SO.sub.4, The residue was purified by silica gel chromatography (PE/EtOAc=10/1) to afford B-10C (2.4 g, impure) as a solid. The impure B-10C (2.4 g) was purified by preparative HPLC (column: Phenomena Luna C18 250*50 mm*10 um, gradient: 20-100% B (A=water (0.05% HCl)-ACN, B=acetonitrile), flow rate: 100 mL/min, 25° C.) to obtain B-10C (1.6 g, 30%) as a foam.

[1879] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.91-7.89 (m, 2H), 7.67-7.64 (m, 1H), 7.59-7.55 (m, 2H), 3.16-3.08 (m, 1H), 3.02-2.94 (m, 1H), 2.08-2.02 (m, 2H), 1.91-1.78 (m, 3H), 1.70-1.60 (m, 4H), 1.53-1.01 (m, 13H), 1.07-0.92 (m, 4H), 0.87-0.85 (m, 3H), 0.83 (s, 3H), 0.68-0.63 (m, 1H), 0.61 (m, 3H).

Step 11

[1880] To a solution of n-BuLi (440 μL, 2.5 M in hexane, 1.10 mmol) in THF (0.5 mL) at −65° C. under N.sub.2 was added a suspension of B-10C (200 mg, 0.3698 mmol) in THF (2.5 mL) dropwise. The mixture was stirred for 30 minutes at −65° C. Diisopropylamine (111 mg, 1.10 mmol) was added at −65° C. After that, (R)-2-methyloxirane (32.2 mg, 0.5547 mmol) was added drop wise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually and stirred at 25° C. for 16 hours. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL), extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give B-11C (250 mg, crude) as a solid, which was used directly for the next step.

Step 12

[1881] To a solution of B-11C (250 mg, 0.417 mmol) in MeOH (10 mL) was added Mg powder (398 mg, 16.6 mmol) and added NiCl.sub.2 (100 mg, 0.771 mmol) was added at 60° C. The mixture was stirred at 60° C. for 3 hrs. After cooling, the mixture was quenched with HCl (50 mL, 1M) and ice-water (50 mL) until the reaction became clear and extracted with EtOAc (2×50 mL). The organic layers were washed with brine (2×50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated. The residue was purified by a silica gel column (PE/EtOAc=10/1) to give Compound 112 (33 mg) as a solid.

[1882] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.80-3.76 (m, 1H), 2.08-1.93 (m, 3H), 1.86-1.77 (m, 2H), 1.70-1.62 (m, 3H), 1.54-1.44 (m, 4H), 1.42-1.32 (m, 6H), 1.31-1.22 (m, 5H), 1.20-1.18 (m, 4H), 1.14-0.96 (m, 6H), 0.94-0.86 (m, 4H), 0.85 (s, 3H), 0.70-0.66 (m, 1H), 0.65 (s, 3H).

[1883] LCMS Rt=1.265 min in 2.0 min chromatography, 30-90 AB

[1884] MS ESI calcd. for C.sub.27H.sub.44F.sub.3O [M+H−H.sub.2O].sup.+ 441, found 441.

Example 113. Synthesis of Compound 2113

[1885] ##STR00597##

Step 1

[1886] To a solution of n-BuLi (440 μL, 2.5 M in hexane, 1.10 mmol) in THF (0.5 mL) at −65° C. under nitrogen was added a suspension of B-10 (200 mg, 0.3698 mmol) in THF (2.5 mL) was added drop-wise. The mixture was stirred for 30 minutes at −65° C., followed by the addition of diisopropylamine (111 mg, 1.10 mmol). After that, (S)-2-methyloxirane (32.2 mg, 0.5547 mmol) was added drop-wise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually and stirred at 25° C. for 16 hours. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (30 mL), extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give C-1 (250 mg, crude) as a solid, which was used directly for the next step.

Step 2

[1887] To a solution of C-1 (250 mg, crude) in MeOH (5 mL) was added Mg (202 mg, 8.34 mmol) under nitrogen. After that, the reaction mixture was stirred at 60° C. for 1 h under N.sub.2. Aq. HCl (1M, 10 mL) was added. The mixture was extracted with EtOAc (2×10 mL). The organic layer washed with saturated aqueous NaHCO.sub.3 (10 mL) solution and brine (10 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give a crude product. The crude product was purified by silica gel column (EtOAc/PE=1/10 to 1/5) to give impure Compound 2113 (100 mg) as a solid, which was re-crystallized from DCM/acetonitrile (5 mL, 2/3) at 25° C. to give Compound 2113 (52 mg, 52%) as a solid.

[1888] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.79 (s, 1H), 2.08-1.94 (m, 3H), 1.82-1.79 (m, 2H), 1.66-1.01 (m, 28H), 0.91-0.90 (m, 4H), 0.85 (s, 3H), 0.71-0.65 (m, 4H).

[1889] LCMS Rt=1.267 min in 2.0 min chromatography, 30-90 AB, MS ESI calcd. for C.sub.27H.sub.44F.sub.3O [M+H−H.sub.2O].sup.+ 441, found 441.

Example 114. Synthesis of Compound 3114

[1890] ##STR00598##

Step 1

[1891] To a solution of diisopropylamine (156 mg, 1.55 mmol) in THF (0.5 mL) was added n-BuLi (0.552 mL, 1.38 mmol, 2.5M in hexane) under N.sub.2 at −70° C. Then a solution of B-10 (300 mg, 0.554 mmol) in THF (3 mL) was added slowly. The mixture was stirred at −70° C. for 30 minutes and then (R)-2-(trifluoromethyl)oxirane (A-1) (93.2 mg, 0.831 mmol) was added and the reaction was stirred at 25° C. for 16 hrs. The mixture was quenched with saturated NH.sub.4Cl (30 mL) and extracted with EtOAc (3×10 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated to give crude D-1 (350 mg) as a solid, which was used directly in the next step.

Step 2

[1892] To a solution of D-1 (350 mg, 0.536 moml) in MeOH (30 mL) was added Mg powder (520 mg, 21.4 mmol) at 55° C. The mixture was stirred at 60° C. for 30 minutes under N.sub.2. The mixture was quenched with HCl (50 mL, 1N) until the reaction became clear and extracted with DCM (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give Compound 3114 (65 mg, 24%) as a solid.

[1893] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.93-3.88 (m, 1H), 2.08-2.01 (m, 1H), 2.00-1.92 (m, 3H), 1.85-1.75 (m, 2H), 1.70-1.58 (m, 5H), 1.50-1.32 (m, 8H), 1.31-1.13 (m, 6H), 1.12-0.96 (m, 5H), 0.95-0.92 (m, 4H), 0.90 (s, 3H), 0.70-0.60 (m, 4H).

[1894] LCMS Rt=1.280 min in 2.0 min chromatography, 30-90 AB

[1895] MS ESI calcd. for C.sub.27H.sub.41F.sub.6O [M+H−H.sub.2O].sup.+ 495, found 495.

Example 115. Synthesis of Compound 4115

[1896] ##STR00599##

Step 1

[1897] To a solution of E-1 (200 mg, mmol, preparation described in WO9827106 and U.S. Pat. No. 5,856,535) and CsF (141 mg, 0.99 mmol) in THF (5 mL) was added drop wise TMSCF.sub.3 (150 mg, 0.99 mmol) at 0° C. The mixture was stirred and kept below 10° C. for 10 mins. Then TBAF (2.5 mL, 1 M in THF, 2.5 mmol) was added at 10° C. and the mixture was stirred and kept below 10° C. for 10 mins. The reaction mixture was quenched by adding water (20 mL) and extracted with EtOAc (2×30 mL). The combined organic layers was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum. The residue was purified by silica gel chromatograph (PE/EtOAc=5/1) to afford E-2 (146 mg, 59%) as a solid.

[1898] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.66 (s, 3H), 2.31-2.21 (m, 2H), 2.08-1.92 (m, 2H), 1.82-1.77 (m, 2H), 1.71-1.65 (m, 4H), 1.50-1.20 (m, 14H), 1.10-0.99 (m, 5H), 0.92-0.84 (m, 7H), 0.71-0.67 (m, 1H), 0.64 (s, 3H).

Step 2

[1899] To a solution of E-2 (146 mg, 0.31 mmol) in THF (1.5 mL) was added Ti(i-PrO).sub.4 (87.5 mg, 0.31 mmol) and followed by adding EtMgBr (0.36 mL, 3 M in Et.sub.2O, 1.07 mmol) dropwise at 25° C. After that, the reaction mixture was stirred at 25° C. for 15 min under N.sub.2. The reaction mixture was quenched with saturated aqueous NH.sub.4Cl (20 mL) solution and extracted with EtOAc (3×20 mL). The combined organic layer was washed with brine (50 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give a crude product, which was purified by silica gel column (EtOAc/PE=5/1) to afford Compound 4115 (45 mg, 31%) as a solid.

[1900] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.10-2.03 (m, 1H), 2.01-1.92 (m, 2H), 1.87-1.77 (m, 2H), 1.75 (s, 1H), 1.72-1.60 (m, 3H), 1.54-1.35 (m, 10H), 1.31-0.98 (m, 12H), 0.92 (d, J=6.5 Hz, 3H), 0.85 (s, 3H), 0.74-0.72 (m, 2H), 0.70-0.67 (m, 1H), 0.65 (s, 3H), 0.45-0.43 (m, 2H). MS ESI calcd. For C.sub.28H.sub.46F.sub.3O.sub.2 [M+H].sup.+ 471, found 471.

Example 116. Synthesis of Compound 5116

[1901] ##STR00600##

Step 1

[1902] To a solution of F-1 (see WO 2017007836 for its synthesis) (7 g, 17.3 mmol) in THF (70 mL) was added MeLi (54.0 mL, 86.4 mmol, 1.6 M in diethyl ether) at 0° C. under N.sub.2. The reaction was warmed to 25° C. and stirred at 25° C. for 1 h. The reaction was quenched with ice water (200 mL), filtered and extracted with EtOAc (2×200 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuum to give F-2 (8 g, crude) as a solid.

[1903] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.4-5.3 (m, 1H), 3.6-3.45 (m, 1H), 2.35-2.25 (m, 2H), 2.05-1.9 (m, 2H), 1.88-1.75 (m, 3H), 1.54-1.26 (m, 15H) 1.24-1.15 (m, 6H), 1.15-1.1 (m, 6H), 1.01-0.95 (m, 3H), 0.93-0.91 (m, 4H), 0.7-0.6 (m, 3H).

Step 2

[1904] To a solution of F-2 (2 g, 4.96 mmol) in MeOH/THF (40 mL/10 mL) was added Pd/C (dry, 10%, 0.8 g). After degassing and back-filling with H.sub.2 for three times, the reaction mixture was stirred for 48 h at 50° C. in H.sub.2 atmosphere (50 psi). The mixture was filtered and concentrated in vacuum to give F-3 (2 g, crude) as a solid.

[1905] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.65-3.50 (m, 1H), 2.01-1.85 (m, 1H), 1.84-1.75 (m, 2H), 1.73-1.57 (m, 5H), 1.55-1.50 (m, 8H), 1.49-1.3 (m, 6H) 1.29-1.15 (m, 6H), 1.15-0.95 (m, 9H), 0.94-0.8 (m, 3H), 0.79-0.75 (m, 3H), 0.7-0.5 (m, 4H).

Step 3

[1906] To the solution of F-3 (2.06 g, 5.10 mmol) in DCM (20 mL) was added PCC (2.21 g, 10.2 mmol) at 25° C., The reaction was stirred at 25° C. for 2 h, The mixture was concentrated in vacuum to give crude F-4 (1.6 g), which was purified by column chromatography on silica gel (0-20% of EtOAc in PE) to give F-4 (1.2 g) as a solid.

[1907] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.45-2.25 (m, 3H), 2.20-1.9 (m, 3H), 1.82-1.75 (m, 1H), 1.73-1.6 (m, 1H), 1.57-1.25 (m, 14H), 1.24-1.1 (m, 14H) 1.09-0.95 (m, 3H), 0.94-0.79 (m, 3H), 0.77-0.6 (m, 4H).

Step 4

[1908] To a solution of F-4 (100 mg, 248 μmol) in THF (5 mL) was added TMSCF.sub.3 (176 mg, 1.24 mmol) and TBAF (0.5 mL, 1 M in THF, 0.5 mmol). The mixture was stirred at 10° C. for 1 h. To the mixture was added TBAF (2.48 mL, 1 M in THF, 2.48 mmol). The mixture was stirred at 30° C. for another 2 h. The mixture was concentrated in vacuum. The residue was dissolved in EtOAc (50 mL), washed with water (2×50 mL), brine (50 mL), dried over Na.sub.2SO.sub.4, filtered, concentrated in vacuum to give crude Compound 5116 (80 mg) as a solid. 50 mg of crude Compound 5116 was triturated with acetonitrile (10 mL) at 50° C. Then the precipitate was collected by filtration and concentrated in vacuum to give Compound 5116 (20 mg).

[1909] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.13-1.97 (m, 2H), 1.96-1.8 (m, 1H), 1.79-1.7 (m, 2H), 1.69-1.61 (m, 4H), 1.60-1.25 (m, 15H), 1.24-1.15 (m, 6H), 1.14-0.92 (m, 7H), 0.9-0.8 (m, 3H), 0.79-0.76 (m, 3H), 0.75-0.55 (m, 4H). LCMS Rt=1.305 min in 2 min chromatography, 30-90 AB, MS ESI calcd. For C.sub.28H.sub.46F.sub.3O [M+H−H.sub.2O]+ 455, found 455.

Example 117: Synthesis for 11786

[1910] ##STR00601##

[1911] The synthesis of DA-24-44 can be found in Example 112. The synthesis of epoxide can be found in Example 87.

Synthesis of ST-200-3CF3-E14_1

[1912] ##STR00602##

[1913] To THF (0.5 mL) was added BuLi (0.368 mL, 2.5 M in hexane, 0.922 mmol). A solution of DA-24-4-4 (200 mg, 0.369 mmol) in THF (3 mL) was added at −70° C. The mixture was stirred at −70° C. for 1 h and iPr.sub.2NH (104 mg, 1.03 mmol) was added. After 10 minutes, a solution of 6,6-difluoro-1-oxaspiro[2.5]octane (163 mg, 0.553 mmol, 50% purity) was added at −70° C. The mixture was stirred at −70° C. for another 1 h. The mixture was warmed to 25° C. and stirred for 16 hrs. To the mixture was added NH.sub.4Cl (50 mL, sat. aq.). The mixture was extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give ST-200-3CF3-E14_1 (250 mg, crude) as a solid, which was used directly for the next.

Synthesis of ST-200-3CF3-E14

[1914] ##STR00603##

[1915] To a solution of ST-200-3CF3-E14_1 (250 mg, 0.362 mmol) in MeOH (20 mL) was added Mg powder (349 mg, 14.4 mmol) at 55° C. The mixture was stirred at 60° C. for 16 hrs. The mixture was quenched with HCl (50 mL, 1 N) until the reaction became clear and extracted with DCM (2 10×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give ST-200-3CF3-E14 (22 mg, 11%) as a solid.

[1916] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.10-2.01 (m, 3H), 1.99-1.88 (m, 4H), 1.86-1.72 (m, 2H), 1.70-1.58 (m, 7H), 1.56-1.32 (m, 10H), 1.31-1.13 (m, 7H), 1.11-0.97 (m, 6H), 0.95-0.86 (m, 4H), 0.84 (s, 3H), 0.65 (s, 3H).

[1917] LCMS Rt=1.329 min in 2.0 min chromatography, 30-90 AB, purity 98.9%, the MS did not show molecular ion by different methods.

Example 118: Synthesis for 11828

[1918] ##STR00604##

[1919] The synthesis of DA-24-4_4 can be found in Example 112.

Synthesis of ST-200-3CF3-25E4S_1

[1920] ##STR00605##

[1921] To a solution of n-BuLi (0.46 mL, 2.5 M in hexane, 1.15 mmol) in THF (1 mL) at −70° C. under N.sub.2 a suspension of DA-24-4_4 (250 mg, 0.46 mmol) in THF (4 mL) was added dropwise. The mixture was stirred for 30 minutes at −70° C. A solution of diisopropylamine (116 mg, 1.15 mmol) was added dropwise at −70° C., then a solution of (S)-2-isopropyloxirane(59.6 mg, 0.69 mmol) was added dropwise at −70° C. The mixture was stirred for another 30 min and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 24 hour. The reaction mixture was quenched by saturated NH.sub.4Cl aqueous (5 mL), extracted with EtOAc (3×10 mL). The combined organic phase was washed with brine (30 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to give ST-200-3CF3-25E4S_1 (500 mg, crude) as a solid, which was used directly for the next step.

Synthesis of ST-200-3CF3-25E4S

[1922] ##STR00606##

[1923] To a solution of ST-200-3CF3-25E4S_1 (500 mg, 0.8 mmol) in 50 mL of anhydrous MeOH was added Mg powder (763 mg, 31.8 mmol) and NiCl.sub.2 (1 mg, 0.008 mmol) with stirring under N.sub.2 at 60° C. The reaction mixture was quenched by 2 M HCl (50 mL) until solid was dissolved. The mixture was extracted with EtOAc (3×50 mL). The combined organic layer was washed with Sat. NaHCO.sub.3 (100 mL), brine (100 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0˜10% of EtOAc in PE) to give to give a solid, which was triturated from n-hexane (5 mL) at 25° C. to give ST-200-3CF3-25E4S (102 mg, 68%) as solid.

[1924] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.44-3.29 (m, 1H), 2.11-1.91 (m, 3H), 1.87-1.60 (m, 6H), 1.52-1.32 (m, 10H), 1.31-0.97 (m, 12H), 0.95-0.88 (m, 10H), 0.86-0.81 (m, 3H), 0.73-0.61 (m, 4H).

[1925] LCMS Rt=1.503 min in 2 min chromatography, 30-90AB_2MIN_E, purity 100%, MS ESI calcd. For C.sub.29H.sub.48F.sub.3O [M+H−H.sub.2O].sup.+ 469, found 469.

Example 119: Synthesis for 11934

[1926] ##STR00607##

[1927] The synthesis of DA-24-44 can be found in Example 112.

Synthesis of DA-24-9_1

[1928] ##STR00608##

[1929] To a solution of n-BuLi (552 μL, 2.5 M in hexane, 1.38 mmol) in THF (1 mL) at −65° C. under N.sub.2 was added a suspension of DA-24-4_4 (250 mg, 0.462 mmol) in THF (4 mL) was added dropwise. The mixture was stirred for 30 minutes at −65° C. Diisopropylamine (139 mg, 1.38 mmol) was added at −65° C. (S)-2-(trifluoromethyl)oxirane (154 mg, 1.38 mmol) was added dropwise at −65° C. The mixture was stirred for another 30 minutes and then warmed to 25° C. gradually. The reaction mixture was stirred at 25° C. for 16 hours and quenched with sat. NH.sub.4Cl aq. (50 mL), extracted with EtOAc (3×50 mL). The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated to give DA-24-9_1 (250 mg, crude) as a solid, which was used directly for the next step.

Synthesis of DA-24-9

[1930] ##STR00609##

[1931] To a solution of DA-24-9_1 (250 mg, 0.382 mmol) and NiCl.sub.2 (4.92 mg, 0.038 mmol) in dry methanol (50 mL) was added Mg powder (364 mg, 15.2 mmol) in 4 portions under N.sub.2 with stirring at 50° C. The reaction mixture was stirred at 60° C. for 1 hour. The mixture was quenched with HCl (50 mL, 1N) until the reaction became clear and extracted with EtOAc (3×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated. The residue was purified by flash column (0-15% of EtOAc in PE) to give impure DA-24-9 (65 mg) as a solid, which was triturated from n-hexane (10 mL) at 68° C. for 2 h to give DA-24-9 (23 mg, 12%) as a solid.

[1932] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.95-3.85 (m, 1H), 2.08-1.90 (m, 4H), 1.70-1.58 (m, 7H), 1.50-1.13 (m, 14H), 1.13-0.98 (m, 5H), 0.98-0.85 (m, 4H), 0.85 (s, 3H), 0.70-0.60 (m, 4H).

[1933] HPLC Rt=3.15 min in 8.0 min chromatography, 50-100_AB_1.2 m1_E, purity 100%.

[1934] HRMS ESI calcd. for C.sub.27H.sub.41F.sub.6 [M+H−H.sub.2O]+ 495.3056, found 495.3050.

Example 120: Synthesis of 12055

[1935] ##STR00610##

[1936] The synthesis of DA-24-4_4 can be found in Example 112.

Synthesis of ST-200-3CF3-E9R_1

[1937] ##STR00611##

[1938] To THF (0.5 mL) was added BuLi (0.46 mL, 2.5 M in hexane, 1.15 mmol). A solution of DA-24-4_4 (250 mg, 0.462 mmol) in THF (3 mL) was added at −70° C. After stirring at −70° C. for 1 h, diisopropylamine (130 mg, 1.29 mmol) was added. After 10 minutes, (S)-2-(tert-butyl)oxirane (69.4 mg, 0.693 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 h. The mixture was warmed to 25° C. and stirred for 16 hrs. To the mixture was added NH.sub.4Cl (50 mL, sat. aq.). The mixture was extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give ST-200-3CF3-E9R_1 (250 mg, crude) as a solid, which was used directly for the next step.

Synthesis of ST-200-3CF3-E9R

[1939] ##STR00612##

[1940] To a solution of ST-200-3CF3-E9R_1 (250 mg, 0.390 mmol) in MeOH (20 mL) was added Mg powder (374 mg, 15.6 mmol) at 55° C. The mixture was stirred at 60° C. for 16 hrs. The mixture was quenched with HCl (50 mL, 1 M) until the reaction became clear and extracted with DCM (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated. The residue was purified by flash column (0-10% of EtOAc in PE) to give ST-200-3CF3-E9R (150 mg, 77%) as a solid. This material was converted to the benzoate ester to facilitate purification of the compound as shown in the next step.

[1941] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.22-3.15 (m, 1H), 2.10-1.93 (m, 3H), 1.88-1.75 (m, 2H), 1.72-1.58 (m, 3H), 1.58-0.97 (m, 23H), 0.97-0.80 (m, 15H), 0.75-0.68 (m, 1H), 0.65 (s, 3H).

Synthesis of ST-200-3CF3-E9R_Bz

[1942] ##STR00613##

[1943] To a solution of ST-200-3CF3_E9R (180 mg, 0.359 mmol) in DCM (2 mL) was added py (567 mg, 7.18 mmol) and BzCl (251 mg, 1.79 mmol). The mixture was stirred at 25° C. for 2 h. The mixture was washed with NaHCO.sub.3 (5 mL, 10% aq), HCl (5 mL, 2 M), purified by prep-TLC (PE:EtOAc=50:1) to give ST-200-3CF3-E9R_Bz (150 mg) as a solid. The crude ST-200-3CF3-E9R_Bz (150 mg) was purified by SFC (Instrument: SFC-14; Column: AD (250 mm*30 mm, 10 um); Condition: 0.1% NH.sub.3H.sub.2O EtOH; Begin B: 30%; End B: 30%; FlowRate(ml/min): 60ML/MIN; Injections: 180) to give pure ST-200-3CF3-E9R_Bz (100 mg) as an oil.

Synthesis of ST-200-3CF3-E9R

[1944] ##STR00614##

[1945] To a solution of ST-200-3CF3-E9R_Bz (100 mg, 0.165 mmol) in THF (1 mL) was added MeOH (0.5 mL), water (0.5 mL) and LiOH.H.sub.2O (69.2 mg, 1.65 mmol), The mixture was stirred at 50° C. for 72 h. To the mixture was added water (2 mL). The mixture was extracted with EtOAc (5 mL). The organic layer was separated, concentrated in vacuum, purified by silica gel column (PE/DCM/EtOAc=40/1/1 to 20/1/1) to give ST-200-3CF3-E9R (60 mg, 73%) as a solid. The impure was re-crystallized from MeCN (0.5 mL) to give ST-200-3CF3-E9R (22 mg) as a solid.

[1946] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.18 (dd, J=5.2, 10.0 Hz, 1H), 2.10-1.93 (m, 3H), 1.88-1.75 (m, 2H), 1.72-1.58 (m, 4H), 1.55-1.28 (m, 12H), 1.28-0.95 (m, 10H), 0.91 (d, J=6.8 Hz, 3H), 0.89 (s, 9H), 0.85 (s, 3H), 0.75-0.68 (m, 1H), 0.65 (s, 3H).

[1947] HPLC Rt=6.63 min in 8.0 min chromatography, 50-100 AB_E, purity 97.3%.

Example 121: Synthesis for 12156

[1948] ##STR00615##

[1949] The synthesis of DA-24-4_4 can be found in Example 112.

Synthesis of ST-200-3CF3-E9S_1

[1950] ##STR00616##

[1951] To THF (0.5 mL) was added BuLi (0.46 mL, 2.5 M in hexane, 1.15 mmol). A solution of DA-24-4_4 (250 mg, 0.462 mmol) in THF (3 mL) was added at −70° C. After stirring at −70° C. for 1 h, diisopropylamine (130 mg, 1.29 mmol) was added. After 10 minutes. (R)-2-(tert-butyl)oxirane (69.4 mg, 0.693 mmol) was added at −70° C. The mixture was stirred at −70° C. for another 1 h. The mixture was warmed to 25° C. and stirred for 16 hrs. To the mixture was added NH.sub.4Cl (50 mL, sat. aq.). The mixture was extracted with EtOAc (2×30 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated to give ST-200-3CF3-E9S_1 (250 mg, crude) as a solid, which was used directly for the next step.

Synthesis of ST-200-3CF3-E9S

[1952] ##STR00617##

[1953] To a solution of ST-200-3CF3-E9S_1 (250 mg, 0.390 mmol) and NiCl.sub.2 (5.03 mg, 0.039 mmol) in dry methanol (20 mL) was added Mg powder (374 mg, 15.6 mmol) in 4 portions under N.sub.2 with stirring at 50° C. The reaction mixture was stirred at 60° C. for 1 hour. The mixture was quenched with HCl (50 mL, 1 M) until the reaction became clear and extracted with DCM (2×30 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered, concentrated and purified by flash column (0-10% of EtOAc in PE) to give ST-200-3CF3-E9S (120 mg, 61%) as a solid

[1954] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.22-3.12 (m, 1H), 2.10-1.90 (m, 3H), 1.90-1.76 (m, 2H), 1.76-1.58 (m, 4H), 1.58-1.32 (m, 7H), 1.32-0.94 (m, 18H), 0.94-0.88 (m, 9H), 0.85 (s, 3H), 0.65 (m, 4H).

Synthesis of ST-200-3CF3-E9S_Bz

[1955] ##STR00618##

[1956] To a solution of ST-200-3CF3_E9S (120 mg, 0.239 mmol) in DCM (2 mL) was added py (377 mg, 4.77 mmol) and BzCl (167 mg, 1.19 mmol). The mixture was stirred at 25° C. for 2 h. The mixture was washed with NaHCO.sub.3 (5 mL, 10% aq), HCl (5 mL, 2 M), purified by prep-TLC (PE:EtOAc=50:1) to give ST-200-3CF3-E9R_Bz (140 mg) as a solid. The crude ST-200-3CF3-E9S_Bz (140 mg) was purified by SFC (Instrument: SFC-14; Column: AD (250 mm*30 mm, 10 um); Condition: 0.1% NH.sub.3H.sub.2O EtOH; Begin B: 30%; End B: 30%; FlowRate(ml/min): 60ML/MIN; Injections: 60) to give pure ST-200-3CF3-E9S_Bz (100 mg) as an oil.

Synthesis of ST-200-3CF3-E9S

[1957] ##STR00619##

[1958] To a solution of ST-200-3CF3-E9S_Bz (100 mg, 0.165 mmol) in THF (1 mL) was added MeOH (0.5 mL), water (0.5 mL) and LiOH.H.sub.2O (69.2 mg, 1.65 mmol), The mixture was stirred at 50° C. for 72 h. To the mixture was added water (2 mL). The mixture was extracted with EtOAc (5 mL). The organic layer was separated, concentrated in vacuum, purified by silica gel column (PE/DCM/EtOAc=40/1/1 to 20/1/1) to give ST-200-3CF3-E9S (60 mg, 73%) as a solid. The impure was re-crystallized from MeCN (1 mL) to give ST-200-3CF3-E9S (35 mg) as a solid.

[1959] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.22-3.12 (m, 1H), 2.10-1.92 (m, 3H), 1.89-1.78 (m, 2H), 1.72-1.57 (m, 5H), 1.55-1.28 (m, 10H), 1.28-0.93 (m, 11H), 0.91 (d, J=6.4 Hz, 3H), 0.89 (s, 9H), 0.85 (s, 3H), 0.74-0.67 (m, 1H), 0.65 (s, 3H).

[1960] HPLC Rt=6.61 min in 8.0 min chromatography, 50-100 AB_E, purity 100%.

Example 123: Biological Data

[1961] The experiments were conducted as described in Example 2 and the results are shown in Table 2-60.

TABLE-US-00007 TABLE 2-60 Avg Avg Avg Avg EC50 Emax EC50 Emax Compound 2A (nM) 2A (%) 2B (nM) 2B (%) [00620] 4115 91.9 186.5 113.6 195.2 [00621] 5116 101.7 185.1 130.4 213.8 [00622] 3114 382.1 318.0 243.5 293.6 [00623] 112 480.1 92.6 478.0 150.8 [00624] 2113 380.4 402.7 412.8 285.0 [00625] 11786 583.9 152.0 1008.7 151.0 [00626] 11828 146.6 142.3 76.9 114.5 [00627] 11934 729.2 113.4 469.7 70.0 [00628] 12055 506.2 113.8 1134.5 184.7 [00629] 12156 153.9 43.2 2456.4 111.2

Example 125: Synthesis

[1962] ##STR00630##

[1963] To a solution of 2,6-di-tert-butyl-4-methylphenol (17 g, 77.1 mmol) in toluene (50 mL) was added trimethylaluminum (19.2 mL, 2M in toluene) at 10° C. The mixture was stirred at 20° C. for 1 h. This MAD solution was used in the next step directly without analysis.

[1964] To a solution of MAD (77.1 mmol) in toluene (50 mL) was added a solution of (R)-methyl 4-((5S,8R,9S,10S,13R,14S,17R)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl) pentanoate (5 g, 12.8 mmol) in toluene (20 mL) dropwise at −70° C. dropwise under N.sub.2. The mixture was stirred at −70° C. for 1 hour. A solution of EtMgBr (12.7 mL, 3M) was added dropwise at −70° C. The mixture was stirred at −70° C. for another 3 hours. When TLC showed most of starting material was consumed and a new spot was produced, the reaction mixture was quenched with citric acid (150 mL, sat. aq.). The reaction was warmed to 25° C. The organic was separated and concentrated in vacuum. The crude was purified by column chromatography on silica gel (EA:PE=200:1 to 10:1) to give (R)-methyl 4-((3S,5S,8R,9S,10S,13R,14S,17R)-3-ethyl-3-hydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoate (3.8 g) as a solid.

[1965] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.66 (s, 3H), 2.41-2.30 (m, 1H), 2.26-2.15 (m, 1H), 1.94 (td, J=3.3, 12.5 Hz, 1H), 1.90-1.73 (m, 2H), 1.69-1.58 (m, 3H), 1.56-0.84 (m, 28H), 0.82 (s, 3H), 0.64 (s, 4H).

Example 126

Synthesis of DA-23-3_3

[1966] ##STR00631##

[1967] LiAlH.sub.4 (198 mg, 2.54 mmol) was added in three portions to a solution of ST-200-3ET-B12_1 (1.1 g, 2.62 mmol) in THF (10 mL) at 0° C. under N.sub.2. After stirring at 20° C. for 1 hour, the mixture was quenched with water (10 mL) at 0° C., followed by adding HCl (10 mL, 1 mol/L). The aqueous phase was extracted with EtOAc (2×10 mL). The combined organic phase was washed with saturated brine (2×10 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by flash column (0-50% of EtOAc in PE) to give DA-23-3_3 (1 g, 98%) as a solid.

[1968] .sup.1H NMR CDCl.sub.3 400 MHz δ 3.65-3.55 (m, 2H), 1.98-1.92 (m, 1H), 1.88-1.75 (m, 1H), 1.70-1.40 (m, 13H), 1.40-1.19 (m, 7H), 1.19-0.98 (m, 7H), 0.98-0.80 (m, 11H), 0.66-0.61 (m, 4H).

Synthesis of DA-23-3_4

[1969] ##STR00632##

[1970] To a solution of DA-23-3_3 (1 g, 2.55 mmol) in anhydrous DCM (30 mL) was added silica gel (1 g) and PCC (1.09 g, 5.10 mmol). After stirring at 20° C. for 1 hours, the reaction mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography on silica gel (PE/EA=50/1 to 10/1) to give DA-23-3_4 (600 mg, 60%) as a solid.

[1971] .sup.1H NMR CDCl.sub.3 400 MHz δ 9.98-9.97 (m, 1H), 2.50-2.20 (m, 2H), 2.05-1.50 (m, 3H), 1.50-1.19 (m, 15H), 1.19-0.99 (m, 7H), 0.99-0.82 (m, 12H), 0.70-0.55 (m, 4H).

Example 127

[1972] ##STR00633##

[1973] To a solution of compound 1 (100 g, 255 mmol, 1.0 eq) in dry MeOH (500 mL) was added concentrated H.sub.2SO.sub.4 (14 mL). The mixture was heated to reflux overnight and then cooled to room temperature. The mixture was quenched with aq. saturated NaHCO.sub.3 solution (0.5 L) and then evaporated to remove MeOH. The residue mixture was extracted with EtOAc (300 mL×3). The combined organic layers were washed with brine (200 mL), dried over Na.sub.2SO.sub.4 and evaporated to give the product (100 g crude, 96%) as a powder.

[1974] .sup.1H NMR: (400 MHz, CDCl3) δ 4.09-4.02 (m, 1H), 3.66 (s, 3H), 3.63-3.58 (m, 1H), 2.39-2.31 (m, 1H), 2.25-2.15 (m, 1H), 1.97-1.91 (m, 1H), 1.91-1.55 (m, 10H), 1.52-1.02 (m, 14H), 0.95-0.88 (m, 6H), 0.62 (s, 3H).

##STR00634##

[1975] To a solution of compound 2 (250 g, 615 mmol, 1.0 eq) in dry pyridine (0.8 L) was added a solution of TsCl (352 g, 1844 mmol, 3.0 eq) in dry pyridine (200 mL). The mixture was stirred at room temperature for 18 h. Ice chips were added gradually to the mixture, and the precipitated solid was filtered, washed with aq. 10% HCl solution (400 mL×3) and water (400 mL×2), and then evaporated to dryness to give crude product (500 g, crude) as a powder, which was used to next step directly

##STR00635##

[1976] A mixture of compound 3 (250 g crude), CH.sub.3COOK (24 g, 245 mmol, 0.77 eq), water (150 mL) and DMF (900 mL) was heated at reflux for 24 h. The solution was cooled to room temperature, with ice chips added gradually. The precipitated solid was filtered off and washed with water (100 mL×2). The crude solid was purified on silica gel column (PE/EtOAc=8/1) to give compound 4 (40 g, yield 34.3% of two steps) as solid.

[1977] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.32-5.38 (m, 1H), 3.66 (s, 3H), 3.47-3.57 (m, 1H), 2.16-2.41 (m, 4H), 1.93-2.04 (m, 2H), 1.74-1.92 (m, 4H), 1.30-1.59 (m, 9H), 0.90-1.19 (m, 12H), 0.68 (s, 3H)

##STR00636##

[1978] To a solution of compound 4 (33 g, 85 mmol, 1.0 eq) in dry CH.sub.2Cl.sub.2 (700 mL) was added Dess-Martin reagent (72 g, 170 mmol, 2.0 eq) in portions at 0° C. Then the reaction mixture was stirred at room temperature for 1 h. TLC (PE:EA=3:1) showed the starting material was consumed completely. The reaction mixture were quenched with a saturated aqueous solution of NaHCO.sub.3/Na.sub.2S.sub.2O.sub.3=1:3 (250 mL). The organic phase was washed with brine (200 mL×2) and dried over Na.sub.2SO.sub.4, and the solvent was evaporated to afford desired product (35 g, crude), which was used in the next step without further purification.

##STR00637##

[1979] To a solution of MAD (0.42 mol, 3.0 eq) in toluene, freshly prepared by addition of a solution of Me.sub.3Al (210 mL, 0.42 mmol, 2 M in hexane) to a stirred solution of 2,6-di-/c/V-butyl-4-methylphenol (185 g, 0.84 mol) in toluene (200 mL) followed by stirring for 1 h at room temperature, was added dropwise a solution of compound 5 (54 g, 0.14 mol, 1.0 eq) in toluene (200 mL) at −78° C. under nitrogen. Then the reaction mixture was stirred for 30 min, a solution of MeMgBr (140 mL, 0.42 mol, 3.0 eq, 3 M in ether) was added dropwise at −78° C. The reaction mixture was warmed to −40° C. and stirred at this temperature for 3 h. TLC (PE:EA=3:1) showed that the starting material was consumed completely. The mixture was poured into aqueous saturated NH.sub.4Cl solution (100 mL) and extracted with EtOAc (300 mL×2). The combined organic phases were dried over Na.sub.2SO.sub.4, and the solvent was evaporated to afford crude product. The crude product was purified on silica gel chromatography eluted with PE:EA=10:1 to give the pure target (30 g, 53%) as powder.

[1980] .sup.1H NMR: (400 MHz, CDCl3) δ 5.31-5.29 (m, 1H), 3.66 (s, 3H), 2.39-2.33 (m, 2H), 2.24-2.22 (m, 1H), 1.99-1.95 (m, 3H), 1.85-1.68 (m, 4H), 1.59-1.40 (m, 8H), 1.31-1.26 (m, 2H), 1.17-1.01 (m, 11H), 0.93-0.91 (m, 4H), 0.67 (s, 3H).

##STR00638##

[1981] A mixture of compound 7 (32.0 g, 82.35 mmol), N,O-dimethylhydroxylamine (16.07 g, 164.70 mmol), HATU (37.57 g, 98.82 mmol) and Et.sub.3N (46.0 mL, 329.40 mmol) in 500 mL anhydrous CH.sub.2Cl.sub.2 was stirred for 18 h at room temperature. TLC showed the reaction was completed. Then CH.sub.2Cl.sub.2 was added to the mixture and the resulting solution was washed with water, 1 N HCl aqueous, saturated aqueous NaHCO.sub.3 and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated, purified by silica gel (PE:EtOAc=10:1 to 3:1) to afford the target compound 8 (17.0 g, yield: 47.8%) as a solid.

[1982] .sup.1H NMR: (400 MHz, CDCl3) δ 5.31-5.29 (m, 1H), 3.69 (s, 3H), 3.17 (s, 3H), 3.03 (s, 2H), 2.47-2.29 (m, 3H), 2.04-1.68 (m, 7H), 1.60-1.43 (m, 7H), 1.38-1.30 (m, 2H), 1.20-1.08 (m, 6H), 1.03-0.91 (m, 8H), 0.68 (s, 3H).

Other Embodiments

[1983] 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.

[1984] 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.

[1985] 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.

[1986] 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.