Steroidal compounds, methods of making and using the same

Abstract

The application discloses a kind of steroid compounds, its preparation method and application.The steroid compound can have the structure shown in formula II and the like.The compound of the application has SREBP pathway inhibitory activity, and can be used for preventing and / or treating obesity, hyperlipidemia, fatty liver, diabetes, atherosclerosis, cardiovascular and cerebrovascular diseases, liver cancer, skin damage and the like.

Description

Technical Field

[0001] This invention relates to a steroidal compound, its preparation method, and its application. Background Technology

[0002] With changing lifestyles, including increased consumption of high-calorie foods and sugary drinks, and lack of exercise and physical activity, metabolic diseases such as hyperlipidemia, obesity, type 2 diabetes, and fatty liver have become increasingly serious health problems worldwide. Fatty liver, in particular, has become a significant cause of chronic liver disease in Europe, America, and affluent regions of my country. The prevalence of simple hepatic lipid accumulation in the general adult population is 10%–30%, of which 10%–20% are steatohepatitis, with a 10-year incidence of cirrhosis and liver cancer reaching 25%. However, the pathophysiological mechanisms of fatty liver are not yet fully understood, and effective and specific treatments are still lacking in clinical practice. It is known that the accumulation of lipids such as cholesterol and triglycerides in the blood and liver is a major cause of hyperlipidemia, which in turn is a significant contributing factor to atherosclerosis, stroke, and fatty liver disease. Therefore, developing novel drugs targeting lipid metabolism regulatory pathways, with a focus on reducing lipids, is increasingly becoming an important direction in the research and development of new drugs for metabolic diseases.

[0003] Lipid synthesis pathways in mammalian cells are known to be crucial for regulating lipid homeostasis. A key factor regulating cholesterol and fatty acid synthesis is the sterol-regulatory element-binding protein (SREBP), a class of transcription factor proteins. The precursors of this protein are first synthesized in the endoplasmic reticulum (ER). These precursors are then transported to the Golgi apparatus via SREBP cleavage-activating protein (SCAP). After further cleavage by two proteases (Site-1 protease (S1P) and Site-2 protease (S2P)), their N-terminal active domain is released, allowing them to enter the nucleus and function as transcription factors. They bind to the SREBP response element (SRE) in the promoter region of target genes, initiating the expression of downstream genes. The cleavage and maturation of SREBP proteins are strictly regulated by intracellular sterol levels (such as cholesterol and 25-hydroxycholesterol). When cells accumulate sufficient cholesterol in the endoplasmic reticulum, cholesterol binds to SCAP and alters the conformation of SCAP, causing the SCAP-SREBP complex to bind to the protein Insig (an insulin-induced gene), thereby blocking the transport of SREBP to the Golgi apparatus and subsequent SREBP activation. Conversely, an increase in the intranuclear active form of SREBP promotes cellular lipid synthesis. Besides cholesterol, 25-hydroxylcholesterol (25-HC) is another potent endogenous inhibitor of the SREBP pathway. Unlike cholesterol binding to SCAP, 25-HC directly binds to Insig and induces the binding of SCAP to Insig.

[0004] Previous studies have found that inhibiting the SREBP pathway is an effective strategy and method for preventing and / or treating metabolic diseases such as obesity, hyperlipidemia, fatty liver, atherosclerosis, and diabetes, as well as cardiovascular and cerebrovascular diseases, skin lesions, and liver cancer.

[0005] Hyperlipidemia's pathogenesis primarily involves increased lipid synthesis or abnormal lipid transport caused by factors such as diet or gene mutations, leading to excessive accumulation of lipids like cholesterol and fatty acids in the blood. Currently, statins and fibrates are the main lipid-lowering drugs used clinically. Statins, in particular, work by inhibiting cellular cholesterol synthesis while simultaneously promoting reverse cholesterol transport in the blood. This indicates that targeting key factors in cellular lipid synthesis is an important means of effectively reducing lipid levels.

[0006] Currently, there are no approved treatments for fatty liver disease, making it crucial to identify therapeutic targets and develop new, effective therapies. The pathogenesis of fatty liver disease involves multiple risk factors, such as the accumulation of triglycerides in the form of lipid droplets, which may trigger steatosis; and abnormally increased cholesterol and fatty acids in cells, which can cause endoplasmic reticulum stress and mitochondrial dysfunction, leading to cell death, inflammation, and fibrosis. Free cholesterol accumulation has been reported as a key driver in the transformation from simple steatosis to invasive steatohepatitis. Furthermore, establishing a mouse model of fatty liver disease has shown that a simple cholesterol-free, high-fat diet, even after prolonged feeding, only induces steatosis, while adding 1-2% cholesterol to the diet is necessary to induce inflammation and fibrosis. Therefore, lowering cholesterol may be a novel treatment strategy for fatty liver disease. Previous studies have shown that abnormal activation of SREBPs has been found in both patients and mouse models of fatty liver disease; the deletion or knockout of liver-specific Scap in mice can eliminate the activation of all SREBPs, thereby preventing the development of fatty liver and hyperlipidemia. Furthermore, recent studies have shown that endoplasmic reticulum stress-induced aberrant SREBP activation promotes adipogenesis and fatty liver. Therefore, this evidence suggests that reducing hepatic triglyceride and cholesterol levels by inhibiting the SREBP pathway is an effective strategy for preventing and / or treating metabolic disorders, including fatty liver. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to provide new compounds with inhibitory activity against the SREBP pathway.

[0008] This invention provides a compound of formula II or a pharmaceutically acceptable salt thereof:

[0009]

[0010] Among them, R 3d For H;

[0011] R 4a It can be H, F, CH3, CF3 or OH;

[0012] R 4b It can be H, F, OH, or CF3;

[0013] Or, R 4a and R 4b And the carbon atoms connected to them together form

[0014] R 22 for Or R 21 ;

[0015] R 21 -L 1-C(O)R A -L 1 -S(O)2R A -L 1 -R B , -L 2 -R c or -L 3 -R D ;

[0016] Each L 1 and L 2 Independently, it is a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one of the -CH2- portions of -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is optionally replaced by -X-;

[0017] L 3 It is -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5- or -(CH2)6-, wherein one of the -CH2- portions of -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5- and -(CH2)6- is replaced by -Y-;

[0018] Each X is independently -CHR 5a -、-CR 5a R 5b -or-Y-;

[0019] Each R 5a and R 5b Each is independently F and C 1-4 Alkyl or fluorinated C 1-4 alkyl;

[0020] Each Y is independently -O-, -NH-, -N(C) 1-4 alkyl)-, -CH=CH-、 or -CHR g -;

[0021] R g -C 1-4 alkylene-OH;

[0022] Each R A Independently for -NR 21a R 21b ;

[0023] R B -N(R) 21a )-C(O)R 21b or -N(R) 21a )-S(O)2R 21b ;

[0024] R D -OH, -CH2OH, R C -CH(CH3)-OH or -C(CH3)2-OH;

[0025] Each R C Independently for -C(R) 21c (R) 21d -OH, CN, -C(O)R E NH2, C 1-4 Alkoxy, 3-6-membered heterocyclic alkyl or 5-10-membered heteroaryl; the 5-10-membered heteroaryl is unsubstituted or substituted with p Rc;

[0026] R E C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl; wherein the C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl and 3-6 membered heterocyclic alkyl groups are independently unsubstituted or substituted by m R groups. a replace;

[0027] Each R 21a H and C independently 1-6 Alkyl, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl, wherein the C 1-6 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocyclic alkyl groups are independently unsubstituted or substituted by m R groups. a replace;

[0028] Each R 21b Independently for H and C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic alkyl, C 6-10 Aryl or 5-10 heteroaryl, wherein the C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic alkyl, C 6-10 The aryl and 5-10 heteroaryl groups are independently unsubstituted or substituted by q R groups. b replace;

[0029] Or, in -NR 21a R 21b In the middle, R 21a and R 21b Together with the nitrogen atoms connecting them, they form 3-10 membered heterocyclic alkyl groups, wherein the 3-10 membered heterocyclic alkyl groups are unsubstituted or p-type R c replace;

[0030] Each m, p, and q is independently 1, 2, 3, 4, or 5;

[0031] Each R a R b and R c Each of the following is independently F, Cl, OH, COOH, CN, NO2, C 1-4 Alkyl, fluorinated C 1-4 Alkyl, C 1-4 Alkoxy,

[0032] R 21c For H, F, C 1-4 Alkyl, fluorinated C 1-4 Alkyl, C 2-4 alkenyl or C 3-6 cycloalkyl;

[0033] R 21d For F, C 2-4 Alkyl, fluorinated C 1-4 Alkyl, C 2-4 alkenyl, C 3-6 Cycloalkyl, phenyl, 5-6 heteroaryl or -L4-R 21e The phenyl group and the 5-6 heteroaryl group are independently unsubstituted or substituted by j R groups. d replace;

[0034] Or, R 21c and R 21d Together with the carbon atoms that connect them, they form C 3-6 cycloalkyl or 3-6 membered heterocyclic alkyl;

[0035] L 4 C 1-4 Alkylene;

[0036] R 21e For OH, CN, C 1-4 Alkyl, phenyl, or 5-6 heteroaryl, wherein the phenyl and 5-6 heteroaryl groups are independently unsubstituted or substituted by j R d replace;

[0037] Each j is independently 1, 2, 3 or 4;

[0038] Each r is independently 0, 1, 2, 3 or 4;

[0039] Each R d and R f Independently, F, Cl, OH, CN, NO2, C 1-4 Alkyl, fluorinated C 1-4 Alkyl or C 1-4 Alkoxy;

[0040] The heterocyclic alkyl and heteroaryl groups have 1, 2, 3 or 4 heteroatoms independently, and each heteroatom is independently N, O or S;

[0041] In Formula II, the carbon atom marked with * is in the R configuration, S configuration, or a mixture of both; when the carbon atom marked with # is a chiral carbon atom, it is in the R configuration, S configuration, or a mixture of both.

[0042] The compound shown in Formula II is not

[0043] In certain preferred embodiments of the present invention, certain groups in the compounds represented by formula II, XX, XXII, IX, XV, XVII or XVIII or their pharmaceutically acceptable salts are defined as follows, and groups not mentioned are the same as those described in any embodiment of the present invention (hereinafter referred to as "in some embodiments" or "in some preferred embodiments").

[0044] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 3d For H;

[0045] R 4a It can be H, F, CH3, CF3 or OH;

[0046] R 4b It can be H, F, OH or CF3;

[0047] Or, R 4a and R 4b And the carbon atoms connected to them together form

[0048] R 22 for Or R 21 ;

[0049] R 21 -L 1 -C(O)R A -L 1 -S(O)2R A -L 1 -R B , -L 2 -R C or -L 3 -R D ;

[0050] Each L 1 and L 2 Independently, it is a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one of the -CH2- portions of -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is optionally replaced by -X-;

[0051] L 3 It is -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5- or -(CH2)6-, wherein one of the -CH2- portions of -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5- and -(CH2)6- is replaced by -Y-;

[0052] Each X is independently -CHR 5a -、-CR 5a R 5b -or-Y-;

[0053] Each R 5a and R 5b Each is independently F and C 1-4 Alkyl or fluorinated C 1-4 alkyl;

[0054] Each Y is independently -O-, -NH-, -N(C) 1-4 alkyl)-, -CH=CH-、 or -CHR g -;

[0055] R g -C 1-4 alkylene-OH;

[0056] Each R A Independently for -NR 21a R 21b ;

[0057] R B -N(R) 21a )-C(O)R 21b or -N(R) 21a )-S(O)2R 21b ;

[0058] R D -OH, -CH2OH, R C -CH(CH3)-OH or -C(CH3)2-OH;

[0059] Each R C Independently for -C(R) 21c (R) 21d -OH, CN, -C(O)R E NH2, C 1-4 Alkoxy, 3-6-membered heterocyclic alkyl or 5-10-membered heteroaryl; said 5-10-membered heteroaryl is unsubstituted or p-shaped. c replace;

[0060] R E C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl; wherein the C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl and 3-6 membered heterocyclic alkyl groups are independently unsubstituted or substituted by m R groups. a replace;

[0061] Each R 21a Independently for H and C 1-6 Alkyl, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl, wherein the C 1-6 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl groups are either unsubstituted or substituted with m Ra groups;

[0062] Each R 21b Independently for H and C 1-6 Alkyl, C 340 Cycloalkyl, 3-10 membered heterocyclic alkyl, C 6-10 Aryl or 5-10 heteroaryl, wherein the C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic alkyl, C 6-10 The aryl and 5-10 heteroaryl groups are independently unsubstituted or substituted by q R groups. b replace;

[0063] Or, in -NR 21a R 21b In the middle, R 21a and R 21bTogether with the nitrogen atoms connecting them, they form 3-10 membered heterocyclic alkyl groups, wherein the 3-10 membered heterocyclic alkyl groups are unsubstituted or substituted by p Rc atoms;

[0064] Each m, p, and q is independently 1, 2, 3, 4, or 5;

[0065] Each Ra, Rb, and Rc is independently F, Cl, OH, COOH, CN, NO2, C 1-4 Alkyl, fluorinated C 1-4 Alkyl, C1.4 alkoxy,

[0066] R 21c For H, F, C 1-4 Alkyl, fluorinated C 1-4 Alkyl, C 2-4 alkenyl or C 3-6 cycloalkyl;

[0067] R 21d For F, C 2-4 Alkyl, fluorinated C 1-4 Alkyl, C 2-4 alkenyl, C 3-6 Cycloalkyl, phenyl, 5-6 heteroaryl or -L 4 -R 21e The phenyl group and the 5-6 heteroaryl group are independently unsubstituted or substituted by j R groups. d replace;

[0068] Or, R 21c and R 21d Together with the carbon atoms that connect them, they form C 3-6 cycloalkyl or 3-6 membered heterocyclic alkyl;

[0069] L 4 C 1-4 Alkylene;

[0070] R 21e For OH, CN, C 1-4 Alkyl, phenyl, or 5-6 heteroaryl, wherein the phenyl and 5-6 heteroaryl groups are independently unsubstituted or substituted by j R d replace;

[0071] Each j is independently 1, 2, 3 or 4;

[0072] Each r is independently 0, 1, 2, 3 or 4;

[0073] Each R d and R f Independently, F, Cl, OH, CN, NO2, C 1-4 Alkyl, fluorinated C1-4 Alkyl or C 1-4 Alkoxy;

[0074] The heterocyclic alkyl and heteroaryl groups have 1, 2, 3 or 4 heteroatoms independently, and each heteroatom is independently N, O or S;

[0075] In Formula II, the carbon atom marked with * is in the R configuration, S configuration, or a mixture of both; when the carbon atom marked with # is a chiral carbon atom, it is in the R configuration, S configuration, or a mixture of both.

[0076] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 3d For H;

[0077] R 4a It can be H, F, CH3 or OH;

[0078] R 4b It can be H, F, OH or CF3;

[0079] Or, R 4a and R 4b And the carbon atoms connected to them together form

[0080] R 22 for Or R 21 ;

[0081] R 21 -L 1 -C(O)R A -L 1 -S(O)2R A -L 1 -R B , -L 2 -R C or -L 3 -R D ;

[0082] Each L 1 and L 2 Independently, it is a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one of the -CH2- portions of -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is optionally replaced by -X-;

[0083] L 3It is -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5- or -(CH2)6-, wherein one of the -CH2- portions of -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5- and -(CH2)6- is replaced by -Y-;

[0084] Each X is independently -CHR 5a -、-CR 5a R 5b -or-Y-;

[0085] Each R 5a and R 5b Each is independently F and C 1-4 Alkyl or fluorinated C 1-4 alkyl;

[0086] Each Y is independently -O-, -NH-, -N(C) 1-4 alkyl)-, -CH=CH-、 or -CHR g -;

[0087] R g -C 1-4 alkylene-OH;

[0088] Each R A Independently for -NR 21a R 21b ;

[0089] R B -N(R) 21a )-C(O)R 21b ;

[0090] R D -OH, -CH2OH, R C -CH(CH3)-OH or -C(CH3)2-OH;

[0091] Each R C Independently for -C(R) 21c (R) 21d -OH, CN, -C(O)R E NH2, C 1-4 Alkoxy, 3-6-membered heterocyclic alkyl or 5-10-membered heteroaryl; said 5-10-membered heteroaryl is unsubstituted or p-shaped. c replace;

[0092] R E C1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl; wherein the C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl and 3-6 membered heterocyclic alkyl groups are independently unsubstituted or substituted by m R groups. a replace;

[0093] Each R 21a Independently for H and C 1-6 Alkyl, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl, wherein the C 1-6 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocyclic alkyl groups are independently unsubstituted or substituted by m R groups. a replace;

[0094] Each R 21b Independently for H and C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic alkyl, C 6-10 Aryl or 5-10 heteroaryl, wherein the C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic alkyl, C 6-10 The aryl and 5-10 heteroaryl groups are independently unsubstituted or substituted by q R groups. b replace;

[0095] Or, in -NR 21a R 21b In the middle, R 21a and R 21b Together with the nitrogen atoms connecting them, they form 3-10 membered heterocyclic alkyl groups, wherein the 3-10 membered heterocyclic alkyl groups are unsubstituted or p-type R c replace;

[0096] Each m, p, and q is independently 1, 2, 3, 4, or 5;

[0097] Each R a R b and R c Each of the following is independently F, Cl, OH, COOH, CN, NO2, C 1-4 Alkyl, fluorinated C 1-4 Alkyl, C 1-4 Alkoxy,

[0098] R 21c For H, F, C 1-4 Alkyl, fluorinated C 1-4 Alkyl, C2-4 alkenyl or C 3-6 cycloalkyl;

[0099] R 21d For F, C 2-4 Alkyl, fluorinated C 1-4 Alkyl, C 2-4 alkenyl, C 3-6 Cycloalkyl, phenyl, 5-6 heteroaryl or -L 4 -R 21e The phenyl group and the 5-6 heteroaryl group are independently unsubstituted or substituted by j R groups. d replace;

[0100] Or, R 21c and R 21d Together with the carbon atoms that connect them, they form C 3-6 cycloalkyl or 3-6 membered heterocyclic alkyl;

[0101] L 4 C 1-4 Alkylene;

[0102] R 21e For OH, CN, C 1-4 Alkyl, phenyl, or 5-6 heteroaryl, wherein the phenyl and 5-6 heteroaryl groups are independently unsubstituted or substituted by j R d replace;

[0103] Each j is independently 1, 2, 3 or 4;

[0104] Each r is independently 0, 1, 2, 3 or 4;

[0105] Each R d and R f Independently, F, Cl, OH, CN, NO2, C 1-4 Alkyl, fluorinated C 1-4 Alkyl or C 1-4 Alkoxy;

[0106] The heterocyclic alkyl and heteroaryl groups have 1, 2, 3 or 4 heteroatoms independently, and each heteroatom is independently N, O or S;

[0107] In Formula II, the carbon atom marked with * is in the R configuration, S configuration, or a mixture of both; when the carbon atom marked with # is a chiral carbon atom, it is in the R configuration, S configuration, or a mixture of both.

[0108] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a For H or CH3, R 4b For H or OH, or R 4a and R4b And the carbon atoms connected to them together form

[0109] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a For H, R 4b It is OH.

[0110] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a CH3, R 4b It is F.

[0111] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a For F, R 4b It is F.

[0112] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a For H, R 4b It is CF3.

[0113] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a CH3, R 4b It is OH.

[0114] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a For OH, R 4b It is CF3.

[0115] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a and R 4b And the carbon atoms connected to them together form

[0116] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a For H, R 4b It is F.

[0117] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a For CF3, R 4b It is CF3.

[0118] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a and R 4b And the carbon atoms connected to them together form

[0119] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 4a For H, R 4b For H.

[0120] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 22 for

[0121] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 22 For R 21 .

[0122] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 -L 1 -C(O)R A .

[0123] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 -L 1 -S(O)2R A .

[0124] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 -L 1 -R B .

[0125] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R 21a Independently for H and C 1-6 Alkyl or C 3-6 cycloalkyl, wherein the C 1-6 Alkyl and C 3-6 The cycloalkyl group is either unsubstituted or substituted with m Ra.

[0126] In some embodiments, m is 1, 2, or 3 in the compound of formula II as described in any of the preceding embodiments.

[0127] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each Ra is independently F, OH, CN or methoxy.

[0128] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R 21a Independently for H and C 1-4 Alkyl, -C 1-4 alkylene -OH, -C 1-4 Alkylene-OC 1-4 Alkyl, fluorinated C1-4 Alkyl or cyclopropyl.

[0129] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R 21a Independently, H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, Cyclopropyl or trifluoromethyl.

[0130] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R 21b Independently for H and C 1-6 Alkyl, C 6-10 Aryl or 5-10 heteroaryl, wherein the C 1-6 Alkyl, C 6-10 The aryl and 5-10 heteroaryl groups are independently unsubstituted or substituted by q R groups. b replace.

[0131] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R 21b Independent of H, -C 1-4 alkylene -OH, -C 1-4 Alkylene-OC 1-4 Alkyl, fluorinated C 1-4 Alkyl, tetrahydromorpholino, phenyl, pyridyl, thiophene, furanyl, pyrrolyl, pyrazolyl, imidazoleyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiazolyl, thiadiazolyl, pyrimidyl, or benzopyrazolyl, wherein the phenyl, pyridyl, thiophene, furanyl, pyrrolyl, pyrazolyl, imidazoleyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiazolyl, thiadiazolyl, pyrimidyl, and benzopyrazolyl groups are independently unsubstituted or substituted by q R groups. b replace.

[0132] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R 21b Independent of H, -C 1-4 alkylene -OH, -C 1-4 Alkylene-OC 1-4 Alkyl, fluorinated C 1-4 Alkyl, tetrahydromorpholino, phenyl The phenyl, It is either unsubstituted or replaced by q Rb.

[0133] In some embodiments, q is 1, 2, or 3 in the compound of formula II as described in any of the preceding embodiments.

[0134] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each Rb is independently F, Cl, OH, COOH, CN, NO2, methyl, trifluoromethyl, methoxy,

[0135] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R 21b Independently, H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, phenyl,

[0136]

[0137] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R 21b Independently

[0138] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, in -NR 21a R 21b In the middle, R 21a and R 21b Together with the nitrogen atoms that connect them, they form The above It is unreplaced or p R c replace.

[0139] In some embodiments, p is 1, 2, or 3 in the compound of formula II as described in any of the preceding embodiments.

[0140] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R c Each is independently F, OH, CN, C 1-4 Alkyl or fluorinated C 1-4 alkyl.

[0141] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R c Each can be independently F, OH, CN, methyl, or trifluoromethyl.

[0142] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, in -NR 21a R 21b In the middle, R 21a and R 21b Together with the nitrogen atoms that connect them, they form

[0143] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R A Independently for -NH2,

[0144]

[0145] In some preferred embodiments, in the compound of formula II as described in any of the preceding embodiments, each R A Independently

[0146]

[0147]

[0148] In some preferred embodiments, in the compound of formula II as described in any of the preceding embodiments, each R A Independently

[0149] In some preferred embodiments, in the compound of formula II as described in any of the preceding embodiments, -L 1 -C(O)R A for

[0150] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R B for

[0151] In some preferred embodiments, in the compound of formula II as described in any of the preceding embodiments, RB is

[0152] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 -L 2 -R C .

[0153] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, L 2 It is a single key.

[0154] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, L 2 It is -CH2-.

[0155] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, L 2 It is -(CH2)2-.

[0156] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, L 2 It is -(CH2)3-.

[0157] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, L 2 It is -(CH2)4-.

[0158] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, L 2 It is -(CH2)5-.

[0159] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, L 2 It is -(CH2)6-.

[0160] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, each R C Independently for -C(R) 21c (R) 21d )-OH.

[0161] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, when R C -C(R) 21c (R) 21d When )-OH, R 21c For H, R 21d C 2-4 Alkyl, fluorinated C 1-4 Alkyl, C 3-6 Cycloalkyl, phenyl, fluorophenyl, 5-6 membered heteroaryl or -L 4 -R 21e .

[0162] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, when R C -C(R) 21c (R) 21d When )-OH, R 21c For H, R 21d C 2-4 Alkyl, fluorinated C 1-4 Alkyl, C 3-6 Cycloalkyl, phenyl, 5-6 heteroaryl or -L 4 -R 21e In which the phenyl group is optionally surrounded by one or two R atoms. d Replace, each R d Independently, it can be F, OH, CN, or C. 1-4 Alkyl group.

[0163] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, L 4 It is methylene or ethylene.

[0164] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21e For OH, CN, C 1.4 alkoxy, phenyl or

[0165] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C -C(R) 21c (R) 21d )-OH is

[0166] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, when R C -C(R) 21c (R) 21d When )-OH, R 21c For F, R 21d C 2-4 Alkyl (e.g., ethyl).

[0167] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C -C(R) 21c (R) 21d )-OH is

[0168] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, when R C -C(R) 21c (R) 21d When )-OH, R 21c It is CH3 or trifluoromethyl, R 21d C 2-4 Alkyl (e.g., ethyl) or trifluoromethyl.

[0169] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C -C(R) 21c (R) 21d )-OH is

[0170] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, when R C -C(R) 21c (R) 21d When )-OH, R21c C 2-4 Alkyl, C 2-4 alkenyl (e.g.) ) or C 3-6 Cycloalkyl (e.g., cyclopropyl), R 21d C 2-4 Alkyl, C 2-4 alkenyl (e.g.) ) or C 3-6 Cycloalkyl (e.g., cyclopropyl).

[0171] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, when R C -C(R) 21c (R) 21d When )-OH, R 21c Ethyl, n-propyl, isopropyl, Or cyclopropyl, R 21d Ethyl, n-propyl, isopropyl, Or cyclopropyl.

[0172] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C -C(R) 21c (R) 21d )-OH is

[0173]

[0174] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, when R C -C(R) 21c (R) 21d When )-OH, R 21c and R 21d Together with the carbon atoms that connect them, they form C 3-6 Cycloalkyl or 3-6 membered heterocyclic alkyl (e.g. ).

[0175] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, when R C -C(R) 21c (R) 21d When )-OH, R 21c and R 21d And together with the carbon atoms that connect them, they form cyclopropyl, cyclobutyl, or...

[0176] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C -C(R) 21c (R) 21d )-OH is

[0177]

[0178] In some preferred embodiments, in the compound of formula II as described in any of the preceding embodiments, R C -C(R) 21c (R) 21d )-OH is

[0179]

[0180] In some preferred embodiments, in the compound of formula II as described in any of the preceding embodiments, R C -C(R) 21c (R) 21d )-OH is

[0181]

[0182]

[0183] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C For CN.

[0184] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C -C(O)R E .

[0185] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R E C 1-4 Alkyl or C 3-6 Cycloalkyl.

[0186] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R E It can be methyl, ethyl, or cyclohexyl.

[0187] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C It is NH2.

[0188] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C C 1-4 Alkyl groups, such as methoxy groups.

[0189] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C for For example

[0190] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C for For example

[0191] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R c for For example

[0192] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C It is a 5-10 member heteroaryl group, wherein the 5-10 member heteroaryl group is unsubstituted or substituted by p Rc groups.

[0193] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C The 5-10 member heteroaryl in the definition is

[0194] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C for In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R C It is a 3-6 membered heterocyclic alkyl group.

[0195] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R c The 3-6 membered heterocyclic alkyl group in the definition is

[0196] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, -L 2 -R C for

[0197] In some preferred embodiments, in the compound of formula II as described in any of the preceding embodiments, -L 2 -R C for

[0198] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 -L 3 -R D .

[0199] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, L 3 for

[0200] The b end of the above structure and R D connect.

[0201] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R D -OH, -CH2OH, CH(CH3)-OH or -C(CH3)2-OH.

[0202] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 -L 3 -R D for

[0203] In some preferred embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 -L 3 -R D for

[0204]

[0205] In some preferred embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 -L 3 -R D for

[0206] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 for

[0207] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 for

[0208] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 for

[0209] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 for

[0210] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 for

[0211] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 for

[0212] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 -L 1 -C(O)R A -L 1 -R B -L 2 -R C or -L 3 -R D .

[0213] In some implementations, L 1 It is -(CH2)2-.

[0214] In some implementations, each R 21 a independently for C 1-6 Alkyl; each R 21b Independently for C 6-10 Aryl or 5-10 heteroaryl, wherein the C 6-10 The aryl and 5-10 heteroaryl groups are independently unsubstituted or substituted by q Rb.

[0215] In some implementations, q is 1.

[0216] In some implementations, each R b Each is independently represented by F.

[0217] In some implementation schemes, R B -N(R) 21a )-C(O)R 21b .

[0218] In some implementations, L 2 It is -(CH2)3-.

[0219] In some implementation schemes, R C -C(R) 21c (R) 21d )-OH or

[0220] In some implementations, r is 0.

[0221] In some implementation schemes, R 21c For H, R 21d fluorinated C 1-4 Alkyl, C 3-6Cycloalkyl or phenyl, wherein the phenyl group is unsubstituted or has a denoted α-aryl group. d Replace; or, R 21c and R 21d Together with the carbon atoms that connect them, they form C 3-6 Cycloalkyl.

[0222] In some implementations, j is 1.

[0223] In some implementation schemes, R d Independently for F, C1 - 4-alkyl or C 1-4 Alkyl group.

[0224] In some implementations, L 3 It is -(CH2)3-. In some implementations, Y is -CH=CH-.

[0225] In some implementation schemes, R D It is -C(CH3)2-OH.

[0226] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, R 21 for

[0227]

[0228] In some embodiments, in the compound of formula II as described in any of the preceding embodiments, when R 4a Let H be the number of 'R', and R be the number of 'R'. 4b When it is H, R 22 For R 21 .

[0229] In some embodiments, the compound represented by Formula II is a compound represented by Formula II-1, II-2, or II-3.

[0230]

[0231] Among them, R 4a For H or CH3, R 22 for Or R 21 R 21 The definition is as described in any embodiment of this invention.

[0232] In some embodiments, in the compound of Formula II as described in any of the preceding embodiments, the carbon atom marked with * in Formula II has an R configuration.

[0233] In some embodiments, in the compound of Formula II as described in any of the preceding embodiments, the carbon atom marked with * in Formula II has an S configuration.

[0234] In some embodiments, in the compound of Formula II as described in any of the preceding embodiments, the carbon atom marked with * in Formula II is a mixture of S-configuration and R-configuration, for example, S-configuration:R-configuration = 1:1.

[0235] In some embodiments, in the compound of Formula II as described in any of the preceding embodiments, when the carbon atom marked with # in Formula II is a chiral carbon atom, it has the R configuration.

[0236] In some embodiments, in the compound of Formula II as described in any of the preceding embodiments, when the carbon atom marked with # in Formula II is a chiral carbon atom, it has an S configuration.

[0237] In some embodiments, in the compound of Formula II as described in any of the preceding embodiments, the carbon atom marked with # in Formula II is a mixture of S configuration and R configuration, for example, S configuration:R configuration = 1:1.

[0238] In some embodiments, the compound represented by Formula II is any of the following compounds:

[0239]

[0240]

[0241] The present invention also provides a compound of formula IX or a pharmaceutically acceptable salt thereof:

[0242]

[0243] Among them, R 3d For H;

[0244] R 6a For H;

[0245] R 6b It can be H, F, or OH;

[0246] Or, R 6a and R 6b Together with the carbon atoms they are connected to form

[0247] R 22 for Or R 21 ;R 21 The definition is as described in any of the schemes of Equation II;

[0248] In Formula IX, the carbon atom marked with * is in the R configuration, S configuration, or a mixture of both; when the carbon atom marked with # is a chiral carbon atom, it is in the R configuration, S configuration, or a mixture of both.

[0249] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, R 22 for

[0250] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, R 22 For R 21 ;R 21 The definition is as described in any of the schemes of Equation II.

[0251] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, R 22 for

[0252] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, R 6a For H, R 6b For H.

[0253] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, R 6a For H, R 6b It is F.

[0254] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, R 6a For H, R 6b It is OH.

[0255] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, the carbon atom marked with * has the R configuration.

[0256] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, the carbon atom marked with * has an S configuration.

[0257] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, the carbon atom marked with * is a mixture of S and R configurations, for example, S configuration:R configuration = 1:1.

[0258] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, the carbon atom marked with # has an R configuration.

[0259] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, the carbon atom marked with # has an S configuration.

[0260] In some embodiments, in the compound of formula IX as described in any of the preceding embodiments, the carbon atom marked with # is a mixture of S configuration and R configuration, for example, S configuration:R configuration = 1:1.

[0261] In some embodiments, the compound represented by Formula IX is the compound represented by Formula IX-1:

[0262]

[0263] Among them, R 6a and R 6b As defined in any embodiment of the present invention, preferably, R 6a For H, R 6b For H.

[0264] In some embodiments, the compound represented by Formula IX is any of the following compounds:

[0265]

[0266] The present invention also provides a compound of formula XV or a pharmaceutically acceptable salt thereof:

[0267]

[0268] Among them, R 3d For H;

[0269] R 22 for Or R 21 ;R 21 The definition is as described in Equation II;

[0270] R 7a It can be H, F, CH3 or CF3;

[0271] In formula XV, the carbon atom marked with * is in the R configuration, S configuration, or a mixture of both.

[0272] In some embodiments, in the compound represented by formula XV as described in any of the preceding embodiments, R 7a It is CH3.

[0273] In some embodiments, in the compound represented by formula XV as described in any of the preceding embodiments, R 22 for

[0274] In some embodiments, in the compound represented by formula XV as described in any of the preceding embodiments, R 22 For R 21 ;R 21 The definition is as described in any of the schemes of Equation II.

[0275] In some embodiments, in the compound represented by formula XV as described in any of the preceding embodiments, R 22 for

[0276] In some embodiments, in the compound represented by formula XV as described in any of the preceding embodiments, the carbon atom marked with * has the R configuration.

[0277] In some embodiments, in the compound represented by formula XV as described in any of the preceding embodiments, the carbon atom marked with * has an S configuration.

[0278] In some embodiments, in the compound represented by formula XV as described in any of the preceding embodiments, the carbon atom marked with * is a mixture of S and R configurations, for example, S configuration:R configuration = 1:1.

[0279] In some embodiments, the compound represented by formula XV is the compound represented by formula XV-1:

[0280]

[0281] Among them, R 7a The definition is as described in any embodiment of this invention.

[0282] In some embodiments, the compound represented by formula XV is a compound that is:

[0283]

[0284] The present invention also provides a compound of formula XVII or a pharmaceutically acceptable salt thereof:

[0285]

[0286] Among them, R 3d For H;

[0287] R 4a The definition is as described in any of the schemes of Equation II;

[0288] R 4b The definition is as described in any of the schemes of Equation II;

[0289] R 22 for

[0290] In Formula XVII, the carbon atom marked with * is in the R configuration, S configuration, or a mixture of both; when the carbon atom marked with # is a chiral carbon atom, it is in the R configuration, S configuration, or a mixture of both.

[0291] In some embodiments, in the compound represented by formula XVII as described in any of the preceding embodiments, R 4a It can be H, F, CH3 or OH;

[0292] R 4b It can be H, F, OH or CF3;

[0293] Or, R 4a and R 4b And the carbon atoms connected to them together form

[0294] In some embodiments, in the compound represented by formula XVII as described in any of the preceding embodiments, R 4a For H; R 4b For H.

[0295] In some embodiments, in the compound of formula XVII as described in any of the preceding embodiments, the carbon atom marked with * has the R configuration.

[0296] In some embodiments, in the compound represented by formula XVII as described in any of the preceding embodiments, the carbon atom marked with * has an S configuration.

[0297] In some embodiments, in the compound represented by formula XVII as described in any of the preceding embodiments, the carbon atom marked with * is a mixture of S and R configurations, for example, S configuration:R configuration = 1:1.

[0298] In some embodiments, in the compound represented by formula XVII as described in any of the preceding embodiments, when the carbon atom marked with # in formula XVII is a chiral carbon atom, it has the R configuration.

[0299] In some embodiments, in the compound represented by formula XVII as described in any of the preceding embodiments, when the carbon atom marked with # in formula XVII is a chiral carbon atom, it has an S configuration.

[0300] In some embodiments, in the compound represented by formula XVII as described in any of the preceding embodiments, the carbon atom marked with # in formula XVII is a mixture of S configuration and R configuration, for example, S configuration:R configuration = 1:1.

[0301] In some embodiments, the compound represented by formula XVII is a compound that is:

[0302]

[0303] The present invention also provides a compound of formula XVIII or a pharmaceutically acceptable salt thereof:

[0304]

[0305] Among them, R 3d For H;

[0306] R 22 for

[0307] In Formula XVIII, the carbon atom marked with * is in the R configuration, S configuration, or a mixture of both.

[0308] In some embodiments, in the compound of formula XVIII as described in any of the preceding embodiments, the carbon atom marked with * has the R configuration.

[0309] In some embodiments, in the compound of formula XVIII as described in any of the preceding embodiments, the carbon atom marked with * has an S configuration.

[0310] In some embodiments, in the compound represented by formula XVIII as described in any of the preceding embodiments, the carbon atom marked with * is a mixture of S and R configurations, for example, S configuration:R configuration = 1:1.

[0311] In some embodiments, the compound represented by formula XVIII is a compound that is:

[0312]

[0313] The present invention also provides a compound of formula XX or a pharmaceutically acceptable salt thereof:

[0314]

[0315] Among them, R 3d For H;

[0316] R 4a The definition is as described in any of the schemes of Equation II;

[0317] R 4b The definition is as described in any of the schemes of Equation II;

[0318] R 7a It can be H, F, CH3 or CF3;

[0319] R 7b It can be F or OH;

[0320] Or, R 7a and R 7b And together with the carbon atoms they are connected to form

[0321] R 22 for Or R 21 ;R 21 The definition is as described in any of the schemes of Equation I;

[0322] In formula XX, the carbon atom marked with * is in the R configuration, S configuration, or a mixture of both; when the carbon atom marked with # is a chiral carbon atom, it is in the R configuration, S configuration, or a mixture of both; when the carbon atom marked with & is a chiral carbon atom, it is in the R configuration, S configuration, or a mixture of both.

[0323] In some implementations, as in the compound represented by formula XX as described above, R 4a For H; R 4b For H.

[0324] In some implementations, as in the compound represented by formula XX as described above, R 7a For H; R 7b It is F.

[0325] In some implementations, as in the compound represented by formula XX as described above, R 7a For F; R 7b It is F.

[0326] In some implementations, as in the compound represented by formula XX as described above, R 7a For H; R 7b It is OH.

[0327] In some implementations, as in the compound represented by formula XX as described above, R 7a and R 7b And together with the carbon atoms they are connected to form

[0328] In some embodiments, in the compound of formula XX as described in any of the preceding embodiments, R 22 for

[0329] In some embodiments, in the compound of formula XX as described in any of the preceding embodiments, the carbon atom marked with * has the R configuration.

[0330] In some embodiments, in the compound represented by formula XX as described in any of the preceding embodiments, the carbon atom marked with * has an S configuration.

[0331] In some embodiments, in the compound represented by formula XX as described in any of the preceding embodiments, the carbon atom marked with * is a mixture of S and R configurations, for example, S configuration:R configuration = 1:1.

[0332] In some embodiments, in the compound represented by formula XX as described in any of the preceding embodiments, the carbon atom marked with # has the R configuration.

[0333] In some embodiments, in the compound represented by formula XX as described in any of the preceding embodiments, the carbon atom marked with # has an S configuration.

[0334] In some embodiments, in the compound represented by formula XX as described in any of the preceding embodiments, the carbon atom marked with # is a mixture of S and R configurations, for example, S configuration:R configuration = 1:1.

[0335] In some embodiments, in the compound represented by formula XX as described in any of the preceding embodiments, the carbon atom labeled & has an R configuration.

[0336] In some embodiments, in the compound represented by formula XX as described in any of the preceding embodiments, the carbon atom labeled & has an S configuration.

[0337] In some embodiments, in the compound represented by formula XX as described in any of the preceding embodiments, the carbon atom labeled & is a mixture of S and R configurations, for example, S configuration:R configuration = 1:1.

[0338] In some embodiments, the compound represented by formula XX is any compound represented by formula XX-1:

[0339]

[0340] Among them, & and R 7a R 7b and R 22 The definitions are as described in any embodiment of this invention.

[0341] In some embodiments, the compound represented by formula XX is any of the following compounds:

[0342]

[0343] The present invention also provides a compound of formula XXII or a pharmaceutically acceptable salt thereof:

[0344]

[0345] Among them, R 3d For H;

[0346] R 4a The definition is as described in any of the schemes of Equation II;

[0347] R 4b The definition is as described in any of the schemes of Equation II;

[0348] R 19 It can be CH2OH, CH2CN, CH2COOH, CH2F, or CHF2;

[0349] R 22 for Or R 21 ;R 21 The definition is as described in any of the schemes of Equation II;

[0350] In Formula XXII, the carbon atom marked with * is in the R configuration, S configuration, or a mixture of both; when the carbon atom marked with # is a chiral carbon atom, it is in the R configuration, S configuration, or a mixture of both.

[0351] In some embodiments, as in the compound represented by formula XXII described above, R 4a For H; R 4b It can be H or OH.

[0352] In some embodiments, as in the compound represented by formula XXII described above, R 4a For H; R 4b For H.

[0353] In some embodiments, as in the compound represented by formula XXII described above, R 4a For H; R 4b It is OH.

[0354] In some embodiments, in the compound represented by formula XXII as described in any of the preceding embodiments, R 22 for

[0355] In some embodiments, in the compound represented by formula XXII as described in any of the preceding embodiments, R 19 It is CH2OH.

[0356] In some embodiments, in the compound represented by formula XXII as described in any of the preceding embodiments, the carbon atom marked with * has the R configuration.

[0357] In some embodiments, in the compound represented by formula XXII as described in any of the preceding embodiments, the carbon atom marked with * has an S configuration.

[0358] In some embodiments, in the compound represented by formula XXII as described in any of the preceding embodiments, the carbon atom marked with * is a mixture of S and R configurations, for example, S configuration:R configuration = 1:1.

[0359] In some embodiments, in the compound represented by formula XXII as described in any of the preceding embodiments, the carbon atom marked with # has an R configuration.

[0360] In some embodiments, in the compound represented by formula XXII as described in any of the preceding embodiments, the carbon atom marked with # has an S configuration.

[0361] In some embodiments, in the compound represented by formula XXII as described in any of the preceding embodiments, the carbon atom marked with # is a mixture of S and R configurations, for example, S configuration:R configuration = 1:1.

[0362] In some embodiments, the compound represented by formula XXII is a compound as shown in XXII-1:

[0363]

[0364] Among them, # and R 4a R 4b and R 19 The definition is as described in any embodiment of this invention.

[0365] In some embodiments, the compound represented by formula XXII is any of the following compounds:

[0366]

[0367] The compounds of the present invention can be prepared from known starting materials (e.g., lanosterol) using various conventional reaction methods in the art (e.g., hydroxyl protection, double bond ozone oxidation, Wittig reaction, hydrolysis, amide condensation, Grignard reagent addition, reduction, nucleophilic substitution, epoxidation). Exemplary preparation methods are described in the preparation examples of this application.

[0368] For example, the preparation of compound 37 and its analogues can be achieved using lanosterol as a starting material, followed by hydroxyl protection, double bond ozone oxidation, Wittig reaction, hydrolysis, and condensation to obtain compound 37 and its analogues; the reaction route is shown below:

[0369]

[0370] For example, the preparation of compound 101 and its analogues can be achieved using lanosterol as a starting material, followed by hydroxyl protection, double bond ozone oxidation, Wittig reaction, hydrolysis, and Grignard reagent addition reaction; the reaction route is shown below:

[0371]

[0372] For example, compound 80 and its analogues can be prepared from lanosterol as a starting material through hydroxyl protection, double bond ozonolysis, reduction, two-step substitution, and addition reactions; the reaction route is shown below:

[0373]

[0374] For example, the preparation method of compound 125 and its analogues can use lanosterol as a starting material, and proceed through hydroxyl protection, double bond ozone oxidation, Wittig reaction, hydrolysis, epoxidation, and hydrolysis to finally obtain compound 125 and its analogues; the reaction route is shown below:

[0375]

[0376] For example, compound 193 and its analogues can be prepared by starting with lanosterol, followed by hydroxyl protection, double bond ozone oxidation, double bond transposition, double bond ozone oxidation, reductive amination, condensation, and hydrolysis to obtain compound 193. The reaction route of the preparation method is shown below:

[0377]

[0378] For example, the preparation method of compound 199 and its analogues uses (22E, 24S)-stigmaster-6(5), 22(23)-dien-3β-ol as the starting material, and proceeds through hydroxyl protection, double bond ozone oxidation, Wittig reaction, hydrolysis, Wittig reaction, double bond reduction, organolithium reagent addition, and epoxidation to obtain product 199. The reaction route of the preparation method is shown below:

[0379]

[0380] The present invention also provides a pharmaceutical composition comprising the compound as described above or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient.

[0381] The present invention also provides the use of the compound as described above or a pharmaceutically acceptable salt thereof or a pharmaceutical composition as described above in the preparation of a medicament for the prevention and / or treatment of a disease, wherein the disease is obesity, hyperlipidemia, fatty liver, diabetes, atherosclerosis, cardiovascular and cerebrovascular diseases, liver cancer or skin lesions.

[0382] The present invention also provides the use of the compound as described above or a pharmaceutically acceptable salt thereof or a pharmaceutical composition as described above in the preparation of a medicament for inhibiting the SREBP pathway.

[0383] The present invention also provides a method for inhibiting the SREBP pathway, comprising administering to a subject an effective amount of the compound as described above or a pharmaceutically acceptable salt thereof.

[0384] The present invention also provides a method for preventing and / or treating a disease, comprising administering to a subject an effective amount of the compound as described above or a pharmaceutically acceptable salt thereof, wherein the disease is obesity, hyperlipidemia, fatty liver, diabetes, atherosclerosis, cardiovascular disease, liver cancer, or skin lesions.

[0385] Definitions and Explanations

[0386] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0387] In this document, the term "substitution" or "substituent" means that a hydrogen atom in a group is replaced by a specified group. When the substitution position is not specified, substitution can occur at any position, but it is only permitted if a stable or chemically viable chemical is formed.

[0388] Examples are given below: The structure represents hydrogen atoms on the benzene ring separated by q R atoms. s Replaced when there are multiple R 8 At that time, each R 8 Same or different.

[0389] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and the Rs in each case have independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0390] When the linking groups listed in this article do not specify their linking direction, the linking direction can be arbitrary, including both left-to-right and right-to-left linking. For example, in -ALB, the linking group L is -CD-. When the linking direction of L is not specified, -ALB includes -ACDB and -ADCB.

[0391] When one of the variables is selected as a single bond, it means that the two groups it connects to are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0392] In this document, the term "alkyl" refers to a saturated straight-chain or branched monovalent hydrocarbon group. C1-C6 alkyl refers to an alkyl group having 1-6 carbon atoms. In some embodiments, C1-C6 alkyl can be C1-C4 alkyl. C1-C4 alkyl includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0393] In this document, the term "alkenyl" refers to a straight-chain or branched monovalent hydrocarbon group containing at least one carbon-carbon double bond. When an alkenyl group contains both saturated and unsaturated carbon atoms, it can be linked to other structures either through the saturated or unsaturated carbon atoms. C2-C4 alkenyl groups represent alkenyl groups having 2, 3, or 4 carbon atoms. Specific examples of alkenyl groups include, but are not limited to, vinyl and allyl groups.

[0394] In this document, the term "alkylene" refers to a saturated straight-chain or branched divalent hydrocarbon group. C1-C4 alkylene refers to alkylene having 1-4 carbon atoms, specifically methylene, ethylene (e.g., -CH2CH2-, -CH(CH3)-), propylene (e.g., -CH2CH2CH2-, -C(CH3)2-, -CH2CH(CH3)-), and butylene (e.g., -CH2CH2CH2CH2-, -CH(CH3)CH(CH3)-, -CH2CH(CH3)CH2-).

[0395] In this document, the term "fluoroalkyl" refers to a group formed by replacing one or more hydrogen atoms in an alkyl group with fluorine, wherein the definition of alkyl is as described above. Examples of fluoroalkyl groups include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl, and pentafluoroethyl.

[0396] In this document, the term "alkoxy" refers to -O-alkyl, where alkyl is defined as previously stated. C1-C4 alkoxy refers to -O-(C1-C4 alkyl), where C1-C4 alkyl is defined as previously stated, that is, C1-C4 alkoxy can specifically be methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, or tert-butoxy.

[0397] In this document, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic cyclic hydrocarbon group (e.g., fused ring, spiro ring, or bridged ring). Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. C 3-10 Specifically, cycloalkyl groups can be C3, C4, C5, C6, C7, C8, C9 ... 10 Cycloalkyl. C 3-6 The cycloalkyl group can specifically be a C3, C4, C5, or C6 cycloalkyl group. In some embodiments, the cycloalkyl group is monocyclic. In some embodiments, the cycloalkyl group is polycyclic (e.g., fused ring, spiro ring, or bridged ring).

[0398] In this document, the term "heterocyclic alkyl" refers to a saturated monocyclic or polycyclic (e.g., fused, spirocyclic, or bridged) cyclic group formed by a carbon atom and at least one heteroatom, wherein the heteroatom is independently selected from N, O, and S. Heterocyclic alkyl groups can be linked to other structures via carbon atoms and heteroatoms on the ring. Examples of heterocyclic alkyl groups include, but are not limited to, those shown below. Tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl. 3-10 membered heterocyclic alkyl groups can specifically be 3, 4, 5, 6, 7, 8, 9, or 10 membered heterocyclic alkyl groups. 3-6 membered heterocyclic alkyl groups can specifically be 3, 4, 5, or 6 membered heterocyclic alkyl groups. In some embodiments, the heterocyclic alkyl group is monocyclic. In some embodiments, the heterocyclic alkyl group is polycyclic (e.g., fused ring, spiro ring, or bridged ring).

[0399] In this article, the term "C" 6-10 "Aryl" refers to phenyl or naphthyl.

[0400] In this document, the term "heteroaryl" refers to an aromatic monocyclic or fused-ring group formed by a carbon atom and at least one heteroatom, wherein the heteroatom is independently selected from N, O, and S. Specifically, 5- to 10-membered heteroaryls can be 5, 6, 7, 8, 9, or 10-membered heteroaryls, such as 5-6-membered heteroaryls or 8- to 10-membered fused heteroaryls. 5- to 6-membered heteroaryls are monocyclic, and specific examples include, but are not limited to, pyrrole, furan, thiophene, oxazole, isoxazole, thiazole, isothiazole, pyrazole, imidazole, pyridine, pyrimidine, and pyrazine. Examples of 8- to 10-membered fused heteroaryls include, but are not limited to, benzopyrrole, benzofuran, benzothiophene, benzoxazole, benzoisoxazole, benzothiazole, benzoisothiazole, benzopyrazole, benzoimidazolium, benzopyrimidine, benzopyrazine, thiazothiazolyl, pyridopyridine, pyridopyrazine, and pyridopyrimidine.

[0401] In this article, the chemical structural formula Indicates the connection position. When Contained in cyclic groups and not specified When the ring atoms are connected, It can attach to any ring atom, but it is only permitted to form a stable or chemically viable chemical compound. For example, include Structures such as...

[0402] In this document, the term "pharmaceutically acceptable salt" refers to a salt formed from a suitable nontoxic organic acid, inorganic acid, organic base, or inorganic base with a compound, which retains the biological activity of the compound. The organic acid may be one or more of the conventional salt-forming organic acids in the art, preferably methanesulfonic acid, p-toluenesulfonic acid, maleic acid, fumaric acid, citric acid, tartaric acid, malic acid, lactic acid, formic acid, acetic acid, propionic acid, trifluoroacetic acid, oxalic acid, succinic acid, benzoic acid, hydroxyethylsulfonic acid, naphthalenesulfonic acid, and salicylic acid. The inorganic acid may be one or more of the conventional salt-forming inorganic acids in the art, preferably hydrochloric acid, sulfuric acid, and phosphoric acid. The organic base may be one or more of the conventional salt-forming organic bases in the art, preferably pyridines, imidazoles, pyrazines, indoles, purines, tertiary amines, and anilines. The tertiary amine organic base is preferably triethylamine and / or N,N-diisopropylethylamine. The aniline organic base is preferably N,N-dimethylaniline. The pyridine organic base is preferably one or more of pyridine, methylpyridine, 4-dimethylaminopyridine, and 2-methyl-5-ethylpyridine. The inorganic base can be any conventional salt-forming inorganic base in the art, preferably one or more of alkali metal hydrides, alkali metal hydroxides, alkali metal alkoxides, potassium carbonate, sodium carbonate, lithium carbonate, cesium carbonate, potassium bicarbonate, and sodium bicarbonate. The alkali metal hydrides are preferably sodium hydride and / or potassium hydride. The alkali metal hydroxides are preferably one or more of sodium hydroxide, potassium hydroxide, and lithium hydroxide. The alkali metal alkoxides are preferably one or more of sodium methoxide, sodium ethoxide, potassium tert-butoxide, and sodium tert-butoxide.

[0403] In chemical structures, wedge-shaped solid line bonds are used. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key Represents the relative configuration of the solid center. (Key) The configuration is not specified, meaning that if configurational isomerism exists in the chemical structure, the bond... It can be Or simultaneously include Two configurations (e.g.) The ratio is 1:1. When the specific configuration of the carbon-carbon double bond is not specified, it can be either E or Z configuration. Stereoisomers can be synthesized using chiral starting materials, prepared by chiral resolution, or resolved using conventional techniques such as, but not limited to, high-performance liquid chromatography (HPLC) using chiral columns.

[0404] In this document, the term "subject" includes any animal, preferably a mammal, and more preferably a human.

[0405] In this document, the term "effective amount" refers to a sufficient amount of a drug or pharmaceutical agent that is non-toxic but achieves the desired effect. The determination of the effective amount varies from person to person, depending on the recipient's age and general condition, as well as the specific active substance. The appropriate effective amount in a case can be determined by a person skilled in the art based on routine testing.

[0406] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0407] The reagents and raw materials used in this invention are all commercially available.

[0408] The positive and progressive effects of this invention are as follows: This invention provides a new class of compounds that have inhibitory activity on the SREBP pathway and can be used to prevent and / or treat diseases such as obesity, hyperlipidemia, fatty liver, diabetes, atherosclerosis, cardiovascular and cerebrovascular diseases, liver cancer, and skin damage. Attached Figure Description

[0409] Figure 1 The results show that 25-hydroxylanosterol inhibits diet-induced weight gain in AMLN-positive mice.

[0410] Figure 2 The effect of 25-hydroxylanosterol on food intake in mice.

[0411] Figure 3 25-hydroxylanosterol reduced the total cholesterol level in the blood of mice.

[0412] Figure 4 25-hydroxylanosterol reduced the total triglyceride level in the blood of mice.

[0413] Figure 5 25-hydroxylanosterol reduced the total cholesterol content in mouse liver.

[0414] Figure 6 25-hydroxylanosterol reduced the total triglyceride content in mouse liver.

[0415] Figure 7 25-hydroxylanosterol reduced the level of aspartate aminotransferase (AST) in the blood of mice.

[0416] Figure 8 25-hydroxylanosterol reduced the level of alanine aminotransferase (ALT) in the blood of mice.

[0417] Figure 9 HE staining results of mouse liver sections: C57BL / 6J CD group.

[0418] Figure 10 HE staining results for mouse liver sections: Ldlr - / -CD set.

[0419] Figure 11 HE staining results for mouse liver sections: Ldlr - / - AMLN group.

[0420] Figure 12 HE staining results for mouse liver sections: Ldlr - / - AMLN+25-HL group.

[0421] Figure 13 HE staining results and NAFLD activity score quantitative statistical results for liver sections of four groups of mice.

[0422] Figure 14 Oil Red O staining results for mouse liver sections: C57BL / 6J CD group.

[0423] Figure 15 Oil Red O staining results for mouse liver sections: Ldlr - / - CD set.

[0424] Figure 16 Oil Red O staining results for mouse liver sections: Ldlr - / - AMLN group.

[0425] Figure 17 Oil Red O staining results for mouse liver sections: Ldlr - / - AMLN+25-HL group.

[0426] Figure 18 The quantitative results of Oil Red O staining of liver sections from four groups of mice were presented.

[0427] Figure 19 Sirius red staining results for mouse liver sections: C57BL / 6J CD group.

[0428] Figure 20 Sirius red staining results for mouse liver sections: Ldlr - / - CD set.

[0429] Figure 21 Sirius red staining results for mouse liver sections: Ldlr - / - AMLN group.

[0430] Figure 22 Sirius red staining results for mouse liver sections: Ldlr - / - AMLN+25-HL group.

[0431] Figure 23 Quantitative results of Sirius red staining on liver sections from four groups of mice.

[0432] Figure 24Immunostaining results of F4 / 80 on mouse liver sections, and imaging results of cholesterol crystals indicated by polarized light imaging: C57BL / 6J CD group.

[0433] Figure 25 Results of F4 / 80 immunostaining of mouse liver sections and imaging results of cholesterol crystals indicated by polarized light: Ldlr - / - CD set.

[0434] Figure 26 Results of F4 / 80 immunostaining of mouse liver sections and imaging results of cholesterol crystals indicated by polarized light: Ldlr - / - AMLN group.

[0435] Figure 27 Results of F4 / 80 immunostaining of mouse liver sections and imaging results of cholesterol crystals indicated by polarized light: Ldlr - / - AMLN+25-HL group.

[0436] Figure 28 The results of F4 / 80 immunostaining quantitative analysis of liver sections from four groups of mice.

[0437] Figure 29 The results of polarized light imaging of liver slices from four groups of mice indicate the quantitative imaging of cholesterol crystals.

[0438] Figure 30 Results of Sudan IV staining of mouse aortic tree: C57BL / 6J CD group.

[0439] Figure 31 Results of Sudan IV staining of mouse aortic tree: Ldlr - / - CD set.

[0440] Figure 32 Results of Sudan IV staining of mouse aortic tree: Ldlr - / - AMLN group.

[0441] Figure 33 Results of Sudan IV staining of mouse aortic tree: Ldlr - / - AMLN+25-HL group.

[0442] Figure 34 Quantitative results of Sudan IV staining for the aortic tree of four groups of mice.

[0443] Figure 35 25-hydroxylanosterol reduces the expression of lipidogenesis-related genes (Hmgcs, Hmgcr, SCD1, FASN) in liver organoids.

[0444] Figure 3625-hydroxylanosterol reduces the expression of fibrosis-related genes (Colla1, αSMA) in liver organoids.

[0445] Figure 37 Bright-field imaging results of mouse liver organoids: solvent control group.

[0446] Figure 38 Bright-field imaging results for mouse liver organoids: obeticholic acid 1μM group.

[0447] Figure 39 Bright-field imaging results for mouse liver organoids: obeticholic acid 3μM group.

[0448] Figure 40 Bright-field imaging results of mouse liver organoids: 25-HL 1μM group.

[0449] Figure 41 Bright-field imaging results of mouse liver organoids: 25-HL 3μM group.

[0450] Figure 42 Nile red staining results for mouse liver organoids: solvent control group.

[0451] Figure 43 Nile red staining results for mouse liver organoids: obeticholic acid 1 μM group.

[0452] Figure 44 Nile red staining results for mouse liver organoids: obeticholic acid 3μM group.

[0453] Figure 45 Nile red staining results for mouse liver organoids: 25-HL 1μM group.

[0454] Figure 46 Nile red staining results for mouse liver organoids: 25-HL 3μM group.

[0455] Figure 47 The results of Nile Red staining quantification of liver organoids from 5 groups of mice.

[0456] Figure 48 Immunofluorescence staining results for αSMA, a marker of fibrosis in mouse liver organoids: solvent control group.

[0457] Figure 49 Immunofluorescence staining results for αSMA, a marker of fibrosis in mouse liver organoids: obeticholic acid 1μM group.

[0458] Figure 50 Immunofluorescence staining results for αSMA, a marker of fibrosis in mouse liver organoids: obeticholic acid 3μM group.

[0459] Figure 51 Immunofluorescence staining results for αSMA, a fibrosis marker protein in mouse liver organoids: 25-HL 1μM group.

[0460] Figure 52 Immunofluorescence staining results for αSMA, a fibrosis marker protein in mouse liver organoids: 25-HL 3μM group.

[0461] Figure 53 The results of immunofluorescence staining quantification of aSMA, a marker protein of fibrosis, in five groups of mouse liver organoids.

[0462] Figure 54 The full two-dimensional NMR spectra of compounds 1-7 are shown.

[0463] Figure 55 This is a magnified two-dimensional NMR spectrum of compound 151-7.

[0464] Figure 56 This is the full two-dimensional NMR spectrum of compound 151.

[0465] Figure 57 This is a magnified view of the two-dimensional NMR spectrum of compound 151.

[0466] Figure 58 Two-dimensional NMR spectrum of compound 203 Figure 2 Spectrum

[0467] Figure 59 This is a magnified view of the two-dimensional NMR spectrum of compound 203. Detailed Implementation

[0468] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0469] In the following examples, 25-hydroxylanosterol (25-HL) refers to the compound of Example 68.

[0470] Animals used in the biological testing example: Adult male C57BL / 6J mice were purchased from Shanghai Silex Biotechnology Co., Ltd., and low-density lipoprotein receptor knockout mice (Ldlr- / - mice, T001464) were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd. Mice were housed under pathogen-free conditions with 12-hour light / dark cycles and free access to water and feed. The gavage solution consisted of 0.5% Tween-80, 0.5% methylcellulose, and 0.9% sodium chloride. The AMLN diet (AMLN, Dyets) contained 40% (kcal percentage) fat (80% of which was trans fat), 22% (by weight) fructose, and 2% (by weight) cholesterol.

[0471] Reagents: Mevalonic acid (41288), paraformaldehyde (P6148), Tween-80 (P8074), methylcellulose (V900506), Oil Red O (O0625), and nuclear staining reagent DAPI were purchased from Sigma-Aldrich. Lovastatin (purity ≥98.5%, HPLC) was purchased from Shanghai Pharmaceutical Valley Co., Ltd. Dulbecco's modified eagle medium (DMEM) for cell culture was purchased from Thermo Scientific, and fetal bovine serum (S1580) was purchased from Biowest. Lipoprotein-free serum (LPDS) was prepared in our laboratory by ultracentrifugation. Obeticholic acid (OCA, CAS Registry No. 459789-99-2, purity 98% (HPLC)). Total cholesterol and total triglyceride kits were purchased from Shanghai Kehua Bioengineering Co., Ltd. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) kits were purchased from Lai Er Biotech. Hematoxylin-eosin staining kits (6765001, 6766010) were purchased from Thermo Scientific. Sirius red staining kit (ab150681) was purchased from Abcam. Sudan IV (A610914) was purchased from Sangon Biotech (Shanghai) Co., Ltd. Nile red (HY-D0718) was purchased from MCE.

[0472] Antibodies: The antibodies used for immunofluorescence staining analysis were as follows: anti-alpha smooth muscle Actin (αSMA, ab7817, 1:500) antibody was purchased from Abcam; FITC-conjugated goat anti-mouse IgG (H+L) (115-095-003) was purchased from Jackson Immunoresearch. Anti-F4 / 80 primary antibody (14-4801-85, Invitrogen, 1:100) and Alexa Fluor Plus 488-conjugated goat anti-rat IgG (A-11006, 1:500) were purchased from Invitrogen.

[0473] Biological Testing Example 1: Cell Culture

[0474] The human hepatocellular carcinoma line Huh-7 / SRE-Luc was grown in a medium containing DMEM, 10% fetal bovine serum, 100 units / ml penicillin, 100 μg / ml streptomycin and 200 μg / ml G418.

[0475] Biological Assay Example 2: SREBP Luciferase Reporter Gene System

[0476] The Huh-7 / SRE-Luc cell line is a human hepatocellular carcinoma cell line that stably expresses LDLR promoter-luciferase and green fluorescent protein (GFP). The LDLR promoter region contains a sterol-regulatory element (SRE), which can effectively and sensitively respond to the regulation of the transcription factor SREBP. GFP signaling serves as an internal control, indicating changes in cell number. Therefore, this cell line can be used to screen for small molecules that regulate the SREBP signaling pathway. In our work, we used this cell line to screen for inhibitors that effectively inhibit the SREBP signaling pathway. Cells were incubated in a sterol-deficient medium (5% delipoproteinized serum, 2 μM lovastatin, 10 μM mevalonic acid) with the corresponding concentrations of the compounds added for 16 hours. After compound treatment, cells were lysed with lysis buffer (E397A, Promega), and luciferase substrate (E1500, Promega) was added. SRE-driven luciferase activity was measured using a BioTek Synergy HTX microplate reader (including but not limited to this type of instrument). The fluorescence intensity of green fluorescent protein (EGFP) was also measured using the aforementioned BioTek microplate reader (including but not limited to this type of instrument) and used as an internal control. The ratio of SRE-driven luciferase activity to the fluorescence intensity of EGFP was used as an indicator of SREBP pathway activity. The test data for each tested compound were analyzed using Prism software to obtain the IC50 of that compound. 50 parameter.

[0477] Biological Testing Example 3: Real-time Quantitative PCR

[0478] Liver or liver organoid samples were homogenized in TRI Reagent (T9424, Sigma), and total RNA was extracted according to the manufacturer's instructions. Equal amounts of RNA template were used to synthesize cDNA using oligodT primers and MLV reverse transcriptase (Promega). Gene quantification data were acquired using a Bio-Rad CFX96 real-time PCR System, and the relative amount of gene mRNA was quantified using the relative CT method. The primer sequences used are shown in the table below:

[0479]

[0480]

[0481] Biological Assay Example 4: Measurement of Serum and Liver Metabolic Parameters

[0482] Mice were euthanized after drug treatment and starved for 4 hours, and their blood and liver were collected. Blood was coagulated and centrifuged at 1500g for 10 minutes at 4°C; the supernatant was serum. Lipids in the liver were extracted using a chloroform / methanol method. The liver was homogenized using a Precelly24 homogenizer, centrifuged at 16000g for 10 minutes at 4°C, and the organic phase was transferred to a new tube, dried under nitrogen, and then dissolved in ethanol. Total cholesterol and triglyceride levels in blood and liver were measured using cholesterol and triglyceride kits (Shanghai Kehua Bioengineering Co., Ltd.). Serum ALT (LE-M0477, Lai Er Bio-tech) and AST (LE-M0568, Lai Er Bio-tech) were measured according to the respective manufacturers' manuals using an analytical system from Sysmex Medical Electronics (Shanghai) Co., Ltd.

[0483] Biological Testing Example 5: Histological Analysis of Liver Sections

[0484] Hematoxylin-eosin staining: The harvested liver sections were fixed in 4% paraformaldehyde at 4°C, embedded in paraffin, and sectioned using a Leica RM2235 microtome to a thickness of 7 μm. After dewaxing and rehydration, the sections were stained using a hematoxylin-eosin staining kit (6765001, 6766010, Thermo Scientific). Images were taken using an Olympus VS 120 slide microscope, and quantification was performed using ImageJ software. Oil Red O staining: The liver sections were embedded in OCT embedding medium (Leica), cut into 7 μm thick sections using a Leica CM1950 cryostat, and stained with Oil Red O (O0625, Sigma). Images were taken using an Olympus VS120 slide microscope, and quantification was performed using ImageJ software. Sirius Red collagen staining: After dewaxing and rehydration, the liver paraffin sections were stained using a Sirius Red staining kit (ab150681, Abcam) according to the manufacturer's instructions. Immunofluorescence staining: Liver sections were embedded in OCT embedding medium (Leica) and cut into 7 μm thick pieces using a cryostat (Leica CM1950). Frozen sections were stained with anti-F4 / 80 rat monoclonal antibody (14-4801-85, Invitrogen, 1:100), Alexa Fluor 488-conjugated goat anti-rat IgG secondary antibody (A-11006, Invitrogen, 1:500), and DAPI (Sigma) 30 was used to specifically stain cell nuclei. After staining and mounting, images were taken using a rotating confocal microscope (Nikon CSU-W1 SoRa), and quantitative analysis was performed using ImageJ software. Polarized light imaging of cholesterol crystals: After F4 / F80 and DAPI staining, frozen sections were photographed using a rotating confocal microscope (Nikon CSU-W1 SoRa) equipped with a polarizing filter, and quantitative analysis was performed using ImageJ software.

[0485] Biological Testing Example 6: Mouse Arterial Tree Isolation and Sudan IV Staining of Atherosclerotic Plaques

[0486] After the mouse drug administration experiment, the aorta was isolated and fixed in 4% PFA. Perivascular adipose tissue was removed under a stereomicroscope using ophthalmic forceps, stained with Sudan IV, and the atherosclerotic plaques were washed with 70% ethanol. Following staining, the aortic tree was imaged using a ZEISS Axio Zoom.V16 stereomicroscope. The atherosclerotic lesions were quantified using ImageJ software.

[0487] Biological Testing Example 7: Preparation and Culture of Mouse Liver Organoids

[0488] C57BL / 6N mice were fed a fatty liver induction diet (TrophicDiet, TP2630052A, containing 10.2% kcal protein, 37.3% kcal carbohydrates, and 52.6% kcal fat) and fructose-containing drinking water (23.1 g fructose and 18.9 g glucose dissolved in 1 L of water, then filtered and sterilized) for 16 weeks at 12 weeks of age to establish a fatty liver model. To generate fatty liver organoids, liver tissue from the fatty liver mice was minced and digested in a digestion buffer at 37°C for 30–60 minutes. The digestion buffer consisted of DMEM / F-12 (Cytiva, SH30023.01), 2.5 mg / mL collagenase D (Roche, COLLD-RO), and 0.1 mg / mL DNase I (Sigma-Aldrich, DN25). Isolated single hepatocytes were filtered through a 70 μm filter and washed once. Cells were collected by centrifugation and resuspended in culture medium mixed with basement membrane extract (BME) (R&D Systems, 3533-010-02) at a ratio of 1:3. The culture medium consisted of AdDMEM / F12, 10 mM Hepes, 1 x Glutamax, 1% pen / strep, 1 x B27, 1 x N2, NAC (1 mM), NIC (10 mM), Gastrino (10 nM), EGF (50 ng / mL), FGF10 (100 ng / mL), A83-01 (5 μM), Rki (10 μM), 10% RSPO1 conditioned medium, 30% Wnt3a conditioned medium, and 5% Noggin conditioned medium. Prior to drug administration, organoids were digested with trypsin (Gibco, Cat#25200072) and resuspended. 24 hours later, using DMSO as a blank control, the organoid medium was changed to contain different concentrations of OCA or 25-HL. The fatty liver liver organoids were divided into 5 groups (1‰ DMSO, 1μM OCA, 3μM OCA, 1μM 25-HL and 3μM 25-HL), and each group was treated with the drug for 72 hours.

[0489] Biological Testing Example 8: Histochemical Staining of Mouse Liver Organoids

[0490] Immunocytochemistry: Organoids were fixed in immunostaining fixative (Beyotime Biotechnology, P0098) overnight at 4°C. They were then washed in PBS and treated with PBS containing 0.5% Triton X-100 at room temperature for 20 minutes. The organoids were then blocked with PBS containing 10% goat serum at room temperature for 1 hour and incubated overnight at 4°C with the primary antibody (Anti-alpha smooth muscle Actin, ab7817, Abcam, 1:500 dilution). The next day, the organoids were washed and co-incubated with the secondary antibody fluorescein (FITC) conjugated with goat anti-mouse IgG (H+L) (Jackson Immunoresearch, 115-095-003). The nuclei were examined using fluorescent shield T cells. M Counterstain with DAPI (Sigma-Aldrich, F6057).

[0491] To observe lipid droplets, organoids were fixed with 4% paraformaldehyde (PFA) for 1 hour and stained with 250 nmol / L Nile Red (MCE, HY-D0718) for 3 minutes at room temperature. The organoids were rinsed twice with PBS before imaging. The stained organoids were observed using a Dragonfly high-speed confocal microscope system (Andor, Dragonfly 200).

[0492] Biological Test Example 9: Inhibitory Effect of the Compounds of the Present Invention on the SREBP Pathway

[0493] The inhibitory effect of the compounds of this invention on the SREBP pathway was tested using the method described in Example 2 of the biological assay. The concentration gradients for each compound were designed as 0.01, 0.03, 0.1, 0.3, 1.0, 3.0, and 10 μM, with DMSO as the control. IC50 values ​​for some compounds were also provided. 50 The values ​​are shown in Table 1.

[0494] Table 1: Activity data of compounds from some examples

[0495]

[0496]

[0497]

[0498]

[0499]

[0500] Biological Test Example 10: 25-Hydroxylanosterol lowers blood lipid levels, alleviates fatty liver and liver damage.

[0501] Elevated blood lipid levels and hepatic lipid accumulation are known to be high-risk factors for fatty liver. This invention further analyzes whether 25-hydroxylanosterol can alleviate the typical symptoms of diet-induced fatty liver in mice: lipid accumulation, liver damage, inflammation, and fibrosis.

[0502] Eight-week-old male C57BL / 6J mice and male Ldlr- / - mice (T001464, Jiangsu Jicui Yaokang Biotechnology Co., Ltd.) were purchased, grouped, and fed different diets and administered different drugs. The mice were randomly divided into four groups of 8-9 mice each: one group consisted of C57BL / 6J wild-type mice fed a basal diet (chow diet, CD) solvent control group; the other three groups were Ldlr- / - mice. - / - Gene knockout mice were divided into three groups: a basal diet (CD) solvent control group, an AMLN diet (AMLNdiet, containing 20% ​​fat, 22% fructose, and 2% cholesterol) solvent control group, and an AMLN diet 25-hydroxylanosterol (25-hydroxylanosterol concentration of 30 mg / kg / day) group. All four groups of mice were administered the diet by gavage once daily, and changes in food intake and body weight were recorded during the treatment. After 8 weeks, blood and liver samples were collected from the mice to analyze the phenotypes of blood lipids, liver lipids, and liver damage.

[0503] The results are as follows Figure 1-8 As shown, where, Figure 1 The statistical results of weekly weight measurements for each group of mice are as follows: Figure 2 The cumulative food intake of mice in each group was statistically analyzed. Figure 3 25-hydroxylanosterol reduced the total cholesterol level in the blood of mice. Figure 4 25-hydroxylanosterol reduced the level of total triglycerides in the blood of mice. Figure 5 25-hydroxylanosterol reduced the total cholesterol content in mouse liver. Figure 6 25-hydroxylanosterol reduced the total triglyceride content in mouse liver. Figure 7 25-hydroxylanosterol reduced serum aspartate aminotransferase (AST) levels in mice. Figure 8 25-hydroxylanosterol reduced the level of alanine aminotransferase (ALT) in the blood of mice.

[0504] Figure 1 and Figure 2 In this study, the p-value was calculated using a two-way ANOVA (Dunnett's multiple comparisons test); * indicates p < 0.05; ns indicates no statistical difference. Figures 3-8In this study, data are expressed as mean ± standard deviation. P-values ​​were determined using one-way ANOVA. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001.

[0505] The results showed that after 8 weeks of continuous administration, Ldlr - / - Mice fed an AMLN diet and simultaneously administered 25-hydroxylanosterol showed significantly lower weight gain compared to the AMLN-sodium-based control group. This indicates that 25-hydroxylanosterol has a good inhibitory effect on AMLN diet-induced weight gain. The pathological features of fatty liver disease mainly include hepatic steatosis, liver damage, inflammatory infiltration, and fibrosis. After 8 weeks of administration to the four groups of mice, we first examined changes in blood lipid levels (total cholesterol, total triglycerides, etc.) and liver lipid levels. Figure 3-4 As shown: Compared with the control group, 25-hydroxylanosterol significantly reduced serum total cholesterol and total triglyceride levels. Meanwhile, as... Figure 5-6 The results showed that 25-hydroxylanosterol significantly reduced total cholesterol and total triglyceride levels in mouse liver. This indicates that 25-hydroxylanosterol has a good effect on lowering blood lipids and liver fat. Figure 7-8 The results showed that serum levels of liver injury markers AST and ALT were significantly reduced by 25-hydroxylanosterol. This indicates that 25-hydroxylanosterol has a good effect on improving liver injury.

[0506] Biological Test Example 11: 25-Hydroxylanosterol Reduces Fatty Liver and Atherosclerosis

[0507] Furthermore, we performed various staining or immunohistochemical operations on liver tissue sections from each group of mice to analyze the phenotypic changes of hepatic lipid accumulation and fatty liver. The results are as follows: Figure 9-29 As shown, where, Figure 9-13 HE staining results and NAFLD activity score quantification results for mouse liver sections: 25-hydroxylanosterol reduced hepatic steatosis and significantly decreased NAFLD activity score; Figure 14-18 Oil Red O staining and quantification results of mouse liver sections: 25-hydroxylanosterol reduced lipid droplets in the liver; Figure 19-23 Sirius red staining and quantitative results of mouse liver sections: 25-hydroxylanosterol reduces fibrosis in the liver; Figure 24-29 Immunostaining and quantification results of F4 / 80 on mouse liver sections, and imaging and quantification results of cholesterol crystallization indicated by polarized light imaging: 25-hydroxylanosterol reduces the aggregation of Kupffer cells and the formation of cholesterol crystals in the liver.

[0508] Figure 9-29Quantitative analysis was performed using ImageJ software, and data are expressed as mean ± standard deviation. P-values ​​were determined using one-way ANOVA: * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001.

[0509] The results are as follows Figure 9-13 As shown by hematoxylin-eosin staining: male Ldlr - / - Mice fed an AMLN diet for 8 weeks showed marked macrovesicular lipid droplets and vesicular degenerated hepatocytes in their livers. Compared with the control group, 25-hydroxylanosterol significantly reduced these phenotypes. Meanwhile, Figure 14-18 Oil Red O staining of liver tissue sections showed that, compared with the control group, 25-hydroxylanosterol significantly reduced lipid droplet accumulation of neutral lipids, including cholesterol and fatty acids, in the liver. These results and Figure 1-8 Consistent with the results showing that 25-hydroxylanosterol reduced liver lipid content, this indicates that 25-hydroxylanosterol effectively improves the liver lipid accumulation phenotype induced by AMLN diet. Furthermore, this invention further analyzed changes in the inflammatory and fibrotic phenotypes associated with fatty liver in the liver. Figure 24-29 Immunofluorescence staining results showed that in mice fed with solvent-controlled Ldlr- / -AMLN diet, F4 / 80-specific staining revealed that Kupffer cells aggregated together and formed crown-like structures around cholesterol crystals. Figure 24-27 (Enlarged image). 25-Hydroxylanosterol significantly reduced the crown-like structures formed by Kupffer cells, indicating that 25-Hydroxylanosterol can significantly reduce inflammatory infiltration in the liver. Polarized light imaging of cholesterol crystals in liver sections showed that in solvent-controlled mice, the crown-like structures formed by Kupffer cells contained a large number of cholesterol crystals. Compared with simple steatosis, these cholesterol crystals and the crown-like structures formed by Kupffer cells are hallmark features of fatty liver. Macrophages are attracted to cholesterol crystals and attempt to clear these residual lipid droplets, similar to the phenomenon described in atherosclerosis. More importantly, 25-Hydroxylanosterol significantly reduced the number of crown-like structures and cholesterol crystals. Figure 24-29 ).at the same time, Figure 19-23 Sirius red staining results showed that 25-hydroxylanosterol significantly reduced the fibrotic phenotype of collagen fiber formation in mouse liver tissue sections. These results suggest that 25-hydroxylanosterol alleviates the symptoms of hepatic steatosis and fatty liver, and may be used for the prevention and / or treatment of diseases such as hyperlipidemia and fatty liver.

[0510] AMLN-fed Ldlr - / - Mice are also a commonly used model for atherosclerosis, so we can study fatty liver and atherosclerosis simultaneously. 8-week-old male Ldlr - / -Mice were fed an AMLN diet and administered the drug via gavage once daily for 8 weeks. After sacrifice, the aorta was isolated and fixed with 4% paraformaldehyde. Perivascular adipose tissue was removed under a stereomicroscope, stained with Sudan IV, and rinsed with 70% ethanol. The aortic tree was imaged using a stereomicroscope (Zeiss Axio ZoomVl6, Germany). Quantitative analysis of atherosclerotic plaques was performed using ImageJ software.

[0511] The results are as follows Figures 30-34 As shown, Figures 30-34 Sudan IV staining and quantification results for mouse aortic tree: 25-hydroxylanosterol reduces the formation of atherosclerotic plaques.

[0512] The results showed that after 8 weeks of continuous administration, the aortic tree of mice in each group was isolated, and aortic Sudan IV lipid-specific staining was performed, such as... Figures 30-34 As shown, compared with the control group, 25-hydroxylanosterol significantly reduced the formation and number of atherosclerotic plaques. 25-hydroxylanosterol has a slowing effect on the formation of atherosclerosis.

[0513] Figure 1-8 to Figure 9-34 These data indicate that 25-hydroxylanosterol reduces elevated blood lipid levels, decreases fat accumulation, cholesterol crystallization, hepatocellular damage, inflammatory infiltration, and fibrosis in the liver, and reduces the formation and number of atherosclerotic plaques. This demonstrates that 25-hydroxylanosterol has good therapeutic effects on hyperlipidemia, fatty liver, and atherosclerosis.

[0514] Example 12 of the biological assay demonstrates, using in vitro liver organoids, that 25-hydroxylanosterol has an inhibitory effect on lipid production and fibrosis in fatty liver.

[0515] To distinguish whether the inhibitory effect of 25-hydroxylanosterol is based on a direct effect on the liver or a systemic effect, we used 3D liver organoids as an in vitro model of fatty liver to analyze the effects of 25-hydroxylanosterol on lipidogenesis and the expression of fibrosis marker molecules in liver organoids. We induced a fatty liver model in C57BL / 6N mice by feeding them a fatty liver-inducing diet (TrophicDiet, TP2630052A, containing 10.2% kcal protein, 37.3% kcal carbohydrate, and 52.6% kcal fat) and fructose-containing drinking water (23.1 g fructose and 18.9 g glucose dissolved in 1 L of water, then filtered and sterilized) for 16 weeks at age 12 weeks. Liver organoids were then isolated from the livers of this model mice and cultured in vitro. Simultaneously, organoids were treated in vitro with obeticholic acid (OCA), an agonist of the fatty X-receptor (FXR). This drug demonstrated good anti-fatty liver effects in phase III clinical trials, and was used as a control drug in this embodiment. Organoids were grouped and treated with the same concentration gradient of obeticholic acid and 25-hydroxylanosterol for 3 days. Organoid RNA was collected, and the expression differences of lipid synthesis-related genes (Hmgcs, Hmgcr; SCD1, FASN) and fibrosis marker molecules (αSMA, Collαl) were analyzed using real-time quantitative PCR.

[0516] The results are as follows Figure 35-53 As shown, where, Figure 35 25-hydroxylanosterol reduced the expression of lipidogenesis-related genes (Hmgcs, Hmgcr, SCD1, FASN) in liver organoids, and the effect was better than that of the control drug obeticholic acid at the same concentration. Figure 36 25-hydroxylanosterol reduced the expression of the liver organoid and fibrosis-associated gene (Collal, αSMA) in patients with liver tissue, and the effect was superior to that of the control drug obeticholic acid at the same concentration. Figure 37-53 Bright-field imaging results of liver organoids, Nile Red staining and quantification results, and immunofluorescence staining and quantification results of fibrosis marker protein αSMA: 25-hydroxylanosterol reduces lipid accumulation in liver organoids and simultaneously reduces the expression of fibrosis marker protein αSMA in liver organoids.

[0517] Figure 35 , Figure 36 Data are expressed as mean ± standard deviation. P-values ​​were determined using one-way ANOVA. ns indicates no statistically significant difference, * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001. Figure 47 and Figure 53Quantitative analysis was performed using ImageJ software. Data are expressed as mean ± standard deviation. P-values ​​were determined using one-way ANOVA: * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001.

[0518] The results showed that, compared to obeticholic acid, which failed to inhibit the expression of lipid synthesis genes, 25-hydroxylanosterol significantly inhibited lipid synthesis-related genes. Figure 35 ) and fibrosis-related genes ( Figure 36 The expression level of ) was measured. Simultaneously, Nile red staining assays were used to detect the accumulation of neutral lipids in organoids, such as... Figure 37-53 The results showed that 25-hydroxylanosterol and OCA reduced lipid accumulation in organoids, and staining data of the fibrosis marker protein αSMA showed a significant reduction in αSMA expression. Figure 35-53 The results indicate that 25-hydroxylanosterol directly reduces hepatic lipid accumulation and fibrosis by inhibiting the expression of adipogenesis and fibrosis genes, while OCA may indirectly inhibit hepatic lipid accumulation by promoting lipid oxidation. These data demonstrate that 25-hydroxylanosterol directly targets and regulates hepatic lipid synthesis pathways and fibrosis genes.

[0519] Example 13: Experiment on the metabolic stability of liver microsomes

[0520] Prepare PBS solution (100 mM), MgCl2 solution (100 mM), and NADPH solution (20 mM) respectively. Then prepare stock solutions of the compound and testosterone (positive control) with dimethyl sulfoxide (DMSO). Dilute with methanol to 100 μM for sample incubation and store at -10 to -30 °C. 12.5 μL of rat liver microsomes (purchased from XenoTech, catalog number: R1000, batch number: 1310030, specification: 20 mg / ml), 432.5 μL of PBS solution (100 mM), 25 μL of NADPH solution (20 mM), and 25 μL of MgCl2 solution (100 mM) were added to a 96-well plate, mixed, and pre-incubated at 37 °C for 5 minutes. 5 μL of substrate (analyte) solution was added to start the reaction. At each set time point (0, 5, 15, 30, 45, and 60 minutes, 0 and 60 minutes for the negative control group), 50 μL of the incubated sample was placed in a stop plate containing 100 μL of ice-cold stop solution, vortexed for 1 minute to inactivate, and stored at -60 to -90 °C for subsequent analysis. The analysis of the analyte and control compound, testosterone, was performed using LC-MS / MS. Analyst software was used for peak integration, calculation, and processing. Both the test compound and the control compound, testosterone, were analyzed using a semi-quantitative method, with the results calculated based on peak area ratio.

[0521] Table 2: Results of rat liver microsomal stability of compounds from some examples

[0522]

[0523]

[0524] Compared with control compound 68, the compound of the present invention exhibits significantly improved stability in rat liver microsomal metabolism, which is significantly superior to compound 68.

[0525] Preparation of key intermediates: Preparation of intermediate II, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-4-formylbutyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanohydro-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester.

[0526]

[0527] Step 1: (1R,3aR,5aR,7S,9aS,11aR)-3a,6,6,9a,11a-pentamethyl-1-[(2R)-6-methylhept-5-en-2-yl]-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (10.00 g, 23.4 mmol, 1.0 eq) was dissolved in DCM (250 mL). After complete dissolution, acetic anhydride (6.7 mL, 71.0 mmol, 3.0 eq), DMAP (0.57 g, 4.7 mmol, 0.2 eq), and TEA (16.3 mL, 117.2 mmol, 5.0 eq) were added sequentially to the reaction system. After the addition was complete, the mixture was stirred at room temperature for 2 hours. After the reaction was complete as monitored by TLC (PE∶EtOAc=5∶1, phosphomolybdic acid plate), it was extracted with methanol (10 mL). The reaction solution was washed once with saturated sodium bicarbonate (~100 mL) and once with water (~100 mL), dried over anhydrous sodium sulfate, concentrated, and methanol (~100 mL) was added when the solution was almost dry. The mixture was stirred in an ice bath for 30 minutes, filtered, and the filter cake was washed with a small amount of methanol. The filter cake was dried to obtain a white solid acetic acid (1R, 3aR, 5). aR,7S,9aS,11aR)-3a,6,6,9a,11a-pentamethyl-1-[(2R)-6-methylhept-5-en-2-yl]-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (I) (9.00 g, 17.3 mmol, purity 90.0%, yield 82.54%). 1H NMR (400MHz, CDCl3): δ4.50 (dd, J=11.5, 4.5Hz, 1H), 2.69 (s, 1H), 2.09-1.87 (m, 8H), 1.77-1 .24 (m, 26H), 1.15 (d, J = 11.4Hz, 3H), 1.00 (s, 3H), 0.93-0.85 (m, 12H), 0.69 (d, J = 2.7Hz, 3H). 13 CNMR (400MHz, CDCl3): δ203.21, 171.00, 134.37, 134.32, 99.99, 80.90, 77.34, 77.02, 76.71, 50.50, 50.30, 49.82, 44.54, 41.14, 37.81, 36. 90, 36.08, 36.03, 35.27, 30.95, 30.78, 28.24, 28.15, 27.91, 26.38, 2 4.23, 24.17, 21.33, 20.98, 19.19, 18.46, 18.40, 18.11, 16.53, 15.78.

[0528] In the second step, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-3a,6,6,9a,11a-pentamethyl-1-[(2R)-6-methylhept-5-en-2-yl]-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (I) (15.00 g, 32.0 mmol, 1.0 eq) was dissolved in DCM (dichloromethane) (300 mL), purged with nitrogen, and the system was cooled to 0 °C in an ice bath. Ozone was then introduced and the mixture was purged for 10 min. The reaction was monitored by TLC (PE∶EtOAc=5∶1, phosphomolybdic acid plate). The product spot was significantly more concentrated than the starting material spot. The gas flow was stopped, the system was replaced with nitrogen and concentrated directly to dryness. The crude product was purified by rapid chromatography (PE∶EtOAc=95∶5 to 90∶10, phosphomolybdic acid plate) to obtain a white solid acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-4-formylbutyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (II) (6.50 g, 13.2 mmol, purity 95.5%, yield 41.3%). 1H NMR (400MHz, CDC13): δ9.77 (t, J=1.9Hz, 1H), 4.50 (dd, J=11.6, 4.5Hz, 1H), 2.51-2.30 (m, 2H), 2.0 9-1.88 (m, 8H), 1.86-1.24 (m, 17H), 1.23-1.12 (m, 2H), 0.99 (d, J=11.2Hz, 3H), 0.93-0.84 (m, 12H).

[0529] Preparation of intermediate VI(5R)-5-[(1R,3aR,5aR,7S,9aS,11aR)-7-hydroxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid

[0530]

[0531] In the first step, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-4-formylbutyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoyl-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (II) (1.50 g, 3.4 mmol, 1.0 eq) was dissolved in dichloromethane (50 mL), and triethylamine (1.41 mL, 10.2 mmol, 3.0 eq) and tert-butyldimethoxytrifluoromethanesulfonate (1.30 g, 5.1 mmol, 1.5 eq) were added sequentially at 0 °C. The reaction solution was stirred at 0°C for 2 hours. TLC (PE:DCM = 3:1, phosphomolybdic acid plate) showed the disappearance of the starting material spot. The reaction solution was concentrated to dryness, and n-hexane (50 mL) was added and stirred for 10 minutes. The mixture was filtered, and the filtrate was evaporated to dryness. DCM (50 mL) was added, and the reaction solution was cooled to -78°C. Ozone was bubbled through the solution for 10 minutes. TLC (PE:DCM = 3:1) showed the disappearance of the starting material spot. PPh3 (0.89 g, 3.4 mmol, 1.0 eq) was added and stirred for 30 minutes. The reaction solution was then directly concentrated and subjected to column chromatography. Purification (PE:EtoAc = 90:10) yielded a white solid acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-1-formylpropyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanohydro-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (III) (300 mg, 0.560 mmol, 16.52%). 1H NMR (400MHz, CDC13) δ9.76 (dd, J=3.3, 1.1Hz, 1H), 4.50 (dd, J=11.6, 4.5Hz, 1H), 2.47 (dd, J=16.4, 2.5Hz, 1H), 2.17 (m, 1H), 2.04 (m, 7H), 1.92 (m, 1H), 1.43 (m, 18H), 0.99 (dd, J=9.3, 2.8Hz, 5H), 0.90 (m, 12H), 0.74 (s, 2H).

[0532] In the second step, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-1-formylpropyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (III) (300 mg, 0.700 mmol, 1.0 eq) was dissolved in dichloromethane (10 mL), and ethyl(triphenylphosphine) acetate (243.81 mg, 0.700 mmol, 1.0 eq) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC (PE:DCM = 3:1, phosphomolybdic acid plate) until complete. The reaction solution was directly concentrated and subjected to silica gel column chromatography (PE:EtOAc = 95:5) to give a white solid (2E,5R)-5-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetyloxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid ethyl ester (IV) (250 mg, 0.401 mmol, 57.30%). 1 H NMR (400MHz, CDCl3) δ6.96 (m, 1H), 5.81 (d, J=15.5Hz, 1H), 4.50 (dd, J=11.5, 4.5Hz, 1H), 4.19 (q, J=7.1Hz, 2H), 1. 99 (m, 8H), 1.63 (m, 12H), 1.31 (m, 6H), 1.17 (m, 3H), 1.00 (s, 3H), 0.92 (t, J=5.3Hz, 4H), 0.87 (m, 10H), 0.69 (m, 3H). 13CNMR (101MHz, CDC13) δ171.01, 148.45, 134.35, 122.47, 80.90, 77.33, 77.21, 77.01, 76.69, 60.12, 50.50, 50.24, 49.86, 44.60, 39.33, 37. 81, 36.91, 36.38, 35.26, 30.80, 28.22, 27.91, 26.92, 26.38, 24.25, 2 4.17, 21.33, 20.96, 19.19, 18.96, 18.11, 16.53, 15.78, 14.29, -0.01.

[0533] Step 3: (2E,5R)-5-[(1R,3aR,5aR,7S,9aS,11aR)-7-hydroxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid ethyl ester (IV) (250 mg, 0.55 mmol, 1.0 eq) was dissolved in methanol (10 mL), and palladium (palladium on carbon) (30 mg, 0.282 mmol) was added. The mixture was stirred for 3 hours under a hydrogen atmosphere. TLC (PE:EtOAc= The reaction was monitored at a ratio of 5:1 (phosphomolybdic acid baking plate). After the reaction was complete, palladium on carbon was filtered off with diatomaceous earth. The filtrate was concentrated to dryness and purified by silica gel column chromatography (PE:EtOAc = 80:20, phosphomolybdic acid baking plate) to obtain a white solid (5R)-5-[(1R,3aR,5aR,7S,9aS,11aR)-7-hydroxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]ethyl hexanoate (V) (200 mg, 0.392 mmol, 71.68%). 1 H NMR (400MHz, CDCl3) δ4.50 (dd, J=11.5, 4.5Hz, 1H), 4.13 (q, J=7.1Hz, 2H), 2.26 (dd, J=15.5, 8.6Hz, 2H), 2.04 (m, 8H), 1.68 (m, 7H), 1.50-1.37 (m, 9H), 1.26 (t, J=7.1Hz, 3H), 1.15 (m, 3H), 1.00 (s, 3H), 0.90 (m, 12H), 0.68 (s, 3H). 13C NMR (101MHz, CDC13) δ173.95, 171.03, 80.95, 77.33, 77.22, 77.02, 76.70, 60.16, 50.52, 50.26, 49.82, 44.49, 37.82, 36.91, 36.21, 35 .69, 35.28, 34.82, 30.96, 30.80, 28.16, 27.92, 26.39, 24.25, 24.18, 21.85, 21.34, 21.00, 19.19, 18.61, 18.13, 16.54, 15.75, 14.28,

[0534] Step 4: Ethyl (5R)-5-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetyloxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (V) (200 mg, 0.40 mmol) was dissolved in ethanol (2 mL), and sodium hydroxide solution (2 mL, 4 mol / L) was added. The reaction mixture was brought to a constant flow and stirred overnight. The reaction was monitored by TLC (PE∶EtOAc=5∶1, phosphomolybdic acid plate). After the reaction was complete, the reaction solution was cooled to room temperature, the pH was adjusted to 1 with 1N hydrochloric acid, and the solution was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (PE:EtOAc = 90:10 to 50:50) to give a white solid (5R)-5-[(1R,3aR,5aR,7S,9aS,11aR)-7-hydroxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid (VI) (150 mg, 0.331 mmol, 82.84%). 1 H NMR (400MHz, CDCl3) δ3.24 (dd, J=11.6, 4.5Hz, 1H), 2.33 (m, 2H), 2.04 (d, J=8.2Hz, 4H), 1.91 (dd, J=13.4, 7.5Hz, 1H), 1.7 0 (m, 6H), 1.50 (m, 7H), 1.19 (m, 7H), 0.99 (d, J=7.8Hz, 5H), 0.91 (t, J=5.9Hz, 3H), 0.87 (s, 3H), 0.81 (s, 3H), 0.69 (s, 3H).

[0535] Preparation of intermediate VII(4R)-4-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]valerate

[0536]

[0537] Acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-4-formylbutyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (II) (3.00 g, 6.78 mmol, 1.0 eq.) was dissolved in acetone (50.0 mL) (it was not completely dissolved and the solution was cloudy). After purging with nitrogen three times, the solution was cooled to 0 °C, and Jones' reagent (7.8 mL, 2.0 M, 2.0 eq.) was slowly added dropwise. After reacting for 5 minutes, the reaction was monitored by TLC (PE∶EA=5∶1, phosphomolybdic acid was used for color development, Rf1=0.72, Rf2=0.34). After the reaction was complete, 15.0 mL of isopropanol was added dropwise at low temperature to quench the reaction. The mixture was stirred for 30 minutes, concentrated, and dissolved in 50.0 mL of dichloromethane. After washing once with 30.0 mL of 1.5N NaHSO3 and 30.0 mL of H2O, and then drying and concentrating, a crude white solid (4R)-4-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetyloxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]valerate (VII) (3.00 g, 4.56 mmol, 67.56%) was obtained. 1 H NMR (400MHz, Chloroform-d) δ4.48 (dd, J=11.6, 4.6Hz, 1H), 2.40 (ddd, J=15.4, 9.9, 5.0Hz, 2H), 2.25 (ddd, J=15.9, 9.5, 6.4Hz, 1H), 2.081.92 (m, 7H), 1.8 41.56 (m, 8H), 1.45 (td, J=13.6, 12.0, 7.2Hz, 3H), 1.31 (dt, J=11.8, 4.3Hz, 2 H), 1.231.07 (m, 3H), 0.98 (s, 3H), 0.88 (q, J=4.7, 3.8Hz, 12H), 0.67 (s, 3H). 13CNMR (100MHz, Chloroform-d) δ179.93, 171.11, 134.35, 134.23, 80.94, 50.44, 50.18, 49.78, 44.49, 37.77, 36.86, 35.98, 35.22, 31.14, 30.99, 30.92, 30.74, 28.04, 27.88, 26.33, 24.20, 24.13, 21.32, 20.95, 19.17, 18.24, 18.08, 16.51, 15.75.

[0538] Preparation of intermediate VIII(4R)-4-[(1R,3aR,5aR,7S,9aS,11aR)-7-hydroxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]valerate

[0539]

[0540] Weigh out (4R)-4-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetyloxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]valeric acid (500 mg, 1.09 mmol, 1.0 eq.), add tetrahydrofuran (20.0 mL), methanol (20.0 mL), and 1M lithium hydroxide aqueous solution (20.0 mL), stir overnight at room temperature, and monitor by TLC. After the reaction was completed, the tetrahydrofuran and methanol were removed by concentration. Water (30.0 mL) was then added to the concentrate, and the mixture was extracted with ethyl acetate (30.0 mL * 3). The organic phases were combined, dried, and concentrated to obtain (4R)-4-[(1R,3aR,5aR,7S,9aS,11aR)-7-hydroxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]valerate (VIII) (350 mg, 77.1%). 1HNMR (400MHz, DMSO-d6) δ11.99 (s, 1H), 4.35 (d, J=5.1Hz, 1H), 3.072.96 (m, 1H), 2.18 (dddd, J=22.7, 15.9, 9.3, 6 .2Hz, 2H), 2.041.84 (m, 5H), 1.731.06 (m, 16H), 0.92 (ddd, J=30.4, 18.0, 4.7Hz, 12H), 0.70 (s, 3H), 0.65 (s, 3H).

[0541] Preparation of intermediate IX(3R)-3-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]butyric acid

[0542]

[0543] Compound Acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-4-formylbutyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoyl-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (III) (2.30 g, 5.2 mmol) 1.0 eq) was dissolved in t-BuOH (tert-butanol) (60 mL), and 15 mL of aqueous solutions of 2-methyl-2-butene (2.19 g, 31.2 mmol, 6.0 eq), sodium dihydrogen phosphate (2.1 g, 15.6 mmol, 3.0 eq), and sodium chlorite (1.41 g, 15.6 mmol, 3.5 eq) were added under ice bath conditions. The reaction mixture was stirred at room temperature for 2 hours. TLC (PE:EtOAc = 5:1, phosphomolybdic acid reagent) showed that the reaction was complete. After concentration, the reaction mixture was extracted with EtOAc (50 mL) and washed with saturated sodium bicarbonate (50 mL × 2). The organic layer was dried with anhydrous sodium sulfate, and the crude product was passed through a column with PE:EtOAc = 3:1 to obtain solid (3R)-3-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]butyric acid (IX) (1.6 g, purity 90%, yield 60.42%). 1H NMR (400MHz, CDCl3) δ4.50 (dd, J=11.6, 4.5Hz, 1H), 2.40 (dd, J=10.2, 5.2Hz, 1H), 2.29 (dd, J=9.5, 6.5Hz, 1H), 2.11-1.87 (m, 8H), 1.83 (dd, J= 10.0, 3.5Hz, 1H), 1.77-1.54(m, 7H), 1.53-1.44(m, 2H), 1.43-1.22(m, 4H), 1.22-1.10(m, 2H), 1.00(s, 3H), 0.97-0.81(m, 12H), 0.69(s, 3H).

[0544] Preparation Examples

[0545] Example 68

[0546] Preparation of compound 68(1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol

[0547]

[0548] In the first step, lanosterol (3.00 g, 7.03 mmol, 1.0 eq) was dissolved in THF (tetrahydrofuran) (273 mL), water (68 mL) was added, and NBS (0.73 g, 4.08 mmol, 0.58 eq) was added at room temperature. The mixture was stirred at 25 °C for 2 hours. TLC (n-hexane∶EtOAc=5∶1, phosphomolybdic acid) showed that the reaction was complete. Extracted three times with dichloromethane, dried over sodium sulfate, evaporated to dryness, and then subjected to column chromatography (PE∶EtOAc=50∶1 to 5∶1). The concentrate yielded (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-5-bromo-6-hydroxy-6-methylhept-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (68-1) (1.3 g, purity 90%, yield 35.3%). 1H NMR (400MHz, CDCl3) δ3.98 (dd, J=21.0, 12.3Hz, 1H), 3.27-3.19 (m, 1H), 2.14 (d, J=6.0Hz, 1H), 2.00 (d, J=11.7Hz, 5H), 1.84-1.61 (m, 6H), 1.58 (s, 9 H), 1.33 (t, J=10.8Hz, 9H), 1.27-1.12 (m, 3H), 1.04 (d, J=12.2Hz, 1H), 0.9 9 (d, J=7.9Hz, 7H), 0.94-0.84 (m, 7H), 0.80 (s, 3H), 0.69 (d, J=2.2Hz, 3H).

[0549] In the second step, (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-5-bromo-6-hydroxy-6-methylhept-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (68-1) (900 mg, 1.72 mmol, 1.0 eq) was dissolved in THF (tetrahydrofuran) (90 mL), and LAH (433 mg, 12.40 mmol, 7.2 eq) was added. After the addition was complete, the temperature was raised to 70°C and refluxed. The mixture was stirred for 2 hours. The reaction was detected by TLC (n-hexane∶EtOAc=5∶1, phosphomolybdic acid). After the reaction was complete, the reaction system was quenched in ice water (10 mL), extracted with DCM (dichloromethane) (30 mL x 3), dried with anhydrous sodium sulfate, and evaporated to dryness to obtain (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (68) (0.4 g, purity 90%, yield 48.7%). 1 H NMR (400MHz, CDCl3) δ3.23 (d, J=8.8Hz, 1H), 2.02 (s, 4H), 1.92 (d, J=8.4Hz, 1H), 1.69 (d, J=24.3Hz, 7H), 1.58 (d, J=11.1Hz, 2H), 1 .52-1.33 (m, 8H), 1.30 (s, 2H), 1.18 (d, J=25.9Hz, 8H), 1.01 (dd, J=22.8, 9.8Hz, 8H), 0.93-0.84 (m, 6H), 0.80 (s, 3H), 0.68 (s, 3H). 13C NMR(101 MHz, CDCl3) δ134.38, 78.97, 77.33, 77.01, 76.69, 71.13, 50.49, 50.38, 49.79, 44.46, 44.39, 38.87, 37.00, 36.72, 36.45 , 35.57, 30.97, 30.82, 29.31, 29.18, 28.23, 27.95, 27.82, 26.48, 24.26, 21.10, 20.99, 19.13, 18.67, 18.24, 15.74, 15.41

[0550] Example 102

[0551] Preparation of compound 102(1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-6-hydroxyoct-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol

[0552]

[0553] Step 1: (1R,3aR,5aR,7S,9aS,11aR)-3a,6,6,9a,11a-pentamethyl-1-[(2R)-6-methylhept-5-en-2-yl]-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (10.00 g, 23.45 mmol, 1.0 eq) was dissolved in DCM (250 mL). After complete dissolution, acetic anhydride (6.7 mL, 70.36 mmol, 3.0 eq), DMAP (0.57 g, 4.69 mmol, 0.2 eq), and TEA (16 mL, 117.25 mmol, 5.0 eq) were added sequentially to the reaction system. After the addition was complete, the mixture was stirred at room temperature for 2 hours. TLC After the reaction (petroleum ether: ethyl acetate = 5:1) was monitored to be complete, it was extracted with methanol (10 mL). The reaction solution was washed once with saturated sodium bicarbonate (~100 mL) and once with water (~100 mL), dried over anhydrous sodium sulfate, concentrated, and when almost dry, methanol (~100 mL) was added. The mixture was stirred in an ice bath for 30 minutes, filtered, and the filter cake was washed with a small amount of methanol. The filter cake was dried to obtain white solid acetic acid (1R, 3aR, 5aR). ,7S,9aS,11aR)-3a,6,6,9a,11a-pentamethyl-1-[(2R)-6-methylhept-5-en-2-yl]-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (I) (9 g, 19.2 mmol, purity 90.0%, yield 73.7%).

[0554] 1 H NMR (400MHz, CDCl3): δ5.10 (s, 1H), 4.50 (dd, J=11.5, 4.6Hz, 1H), 2.08-1.97 (m, 8H), 1.95-1.82 (m, 2H), 1.7 6-1.58(m, 10H), 1.58-1.43(m, 4H), 1.42-1.22(m, 6H), 1.20-1.14(m, 3H), 1.00(s, 3H), 0.95-0.83(m, 15H). 13CNMR (400MHz, CDCl3): δ203.21, 171.00, 134.37, 134.32, 99.99, 80.90, 77.34, 77.02, 76.71, 50.50, 50.30, 49.82, 44.54, 41.14, 37.81, 36. 90, 36.08, 36.03, 35.27, 30.95, 30.78, 28.24, 28.15, 27.91, 26.38, 2 4.23, 24.17, 21.33, 20.98, 19.19, 18.46, 18.40, 18.11, 16.53, 15.78.

[0555] In the second step, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-3a,6,6,9a,11a-pentamethyl-1-[(2R)-6-methylhept-5-en-2-yl]-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (I) (15.00 g, 32.00 mmol, 1.0 eq) was dissolved in DCM (dichloromethane) (300 mL), purged with nitrogen, and the system was cooled to 0 °C in an ice bath. Ozone was then introduced and the mixture was purged for 10 min. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). The product spot was significantly more concentrated than the starting spot. The gas flow was stopped, the system was replaced with nitrogen and concentrated directly to dryness. The crude product was purified by rapid chromatography (petroleum ether:ethyl acetate = 95:5 to 90:10) to obtain a white solid acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-4-formylbutyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (II) (6.50 g, 14.68 mmol, purity 95.5%, yield 43.8%). 1 H NMR (400MHz, CDCl3): δ9.77 (t, J=1.9Hz, 1H), 4.50 (dd, J=11.6, 4.5Hz, 1H), 2.51-2.30 (m, 2H), 2.0 9-1.88 (m, 8H), 1.86-1.24 (m, 17H), 1.23-1.12 (m, 2H), 0.99 (d, J=11.2Hz, 3H), 0.93-0.84 (m, 12H).

[0556] Step 3: (Methoxymethyl)triphenylphosphine chloride (3.87 g, 11.32 mmol, 5.0 eq) was dissolved in anhydrous tetrahydrofuran (40 mL). The system was cooled to 0 °C in an ice-water bath, and NaHMDS (11 mL, 1 mol / L, 5.0 eq) was added. After stirring for 20 minutes, acetic acid (1R, 3aR, 5aR, 7S, 9aS, 11aR)-1-[(2R)-4-formylbutyric acid] [2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (II) (1.00 g, 2.26 mmol, 1.0 eq) was dissolved in anhydrous tetrahydrofuran (10 mL) and added to the reaction solution. The system was brought to room temperature and stirred for 2 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was complete, water (20 mL) was added to the reaction solution, the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 90:10) to give a white solid acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R,4E)-5-methoxypentan-4-en-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,1la-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (37-1) (850 mg, 1.81 mmol, purity 80%, yield 64.1%). 1 H NMR (400MHz, CDCl3) δ6.28 (d, J=12.6Hz, 1H), 5.85 (d, J=6.2Hz, 1H), 4.71 (d, J=12.6Hz, 1H), 4.50 (dd, J=11.5, 4.5Hz, 1H), 4.31 (d, J=6.5Hz, 1 H), 3.58(s, 1H), 3.52-3.47(m, 2H), 2.09-1.88(m, 10H), 1.80-1.24(m, 15H), 1.21-1.11(m, 2H), 1.00(s, 3H), 0.93-0.83(m, 12H), 0.69(s, 3H).

[0557] In the fourth step, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R,4E)-5-methoxypentan-4-en-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (37-1) (700 mg, 1.49 mmol, 1.0 eq) was dissolved in a mixture of dilute hydrochloric acid (5 mol / L, 20 mL) and THF (20 mL). The reaction mixture was heated to 50 °C and stirred for 2 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was complete, the reaction solution was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sulfuric acid, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 95:5 to 85:15) to give a white solid acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-5-formylpentan-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (98-1) (500 mg, 1.09 mmol, purity 80%, yield 58.5%). 1 H NMR (400MHz, CDCl3) δ9.77 (t, J=1.7Hz, 1H), 4.50 (dd, J=11.6, 4.5Hz, 1H), 2.40 (td, J=7.9, 1.7Hz, 2H), 2.12-1.97 (m, 7H), 1.96- 1.85 (m, 1H), 1.78-1.63 (m, 7H), 1.59-1.24 (m, 9H), 1.21-1.06 (m, 3H), 1.00 (s, 3H), 0.90 (dd, J=17.5, 5.5Hz, 12H), 0.69 (s, 3H). 13 C NMR (101MHz, CDCl3) δ202.99, 171.02, 134.43, 134.30, 129.64, 115.29, 80.93, 77.33, 77.02, 76.70, 50.51, 50.24, 49.82, 44.50, 44.35, 37.81, 36.90, 36.31, 35.74, 35.27, 30.96, 30.79, 28.19, 27.92, 26.38, 24.24, 24.18, 21.33, 20.99, 19.19, 18.96, 18.57, 18.12, 16.53, 15.75, -0.00.

[0558] In the fifth step, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-5-formylpentan-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (98-1) (80 mg, 0.18 mmol, 1.0 eq) was dissolved in anhydrous tetrahydrofuran (5 mL), and ethyl magnesium chloride (0.9 mL, 1.80 mmol, 2.0 mol / L, 10 eq) was added under nitrogen protection at 0°. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was complete, the reaction was quenched with ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 90:10) to give a white solid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-6-hydroxyoctyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (102) (45 mg, 0.10 mmol, purity 95%, yield 52.8%). 1 H NMR (400MHz, CDCl3) δ3.52-3.40 (m, 1H), 3.17 (dd, J=11.5, 4.5Hz, 1H), 2.00-1.9 1(m, 4H), 1.88-1.81(m, 1H), 1.68-1.58(m, 5H), 1.54-1.42(m, 6H), 1.35-1.22(m, 7H), 1.20-1.05 (m, 4H), 0.98 (dd, J=12.6, 2.1Hz, 2H), 0.92 (d, J=7.8Hz, 6H), 0.8 8 (t, J=7.5Hz, 3H), 0.83 (d, J=6.4Hz, 3H), 0.81 (s, 3H), 0.74 (s, 3H), 0.62 (s, 3H). 13 C NMR(101 MHz, CDCl3) δ128.14, 128.11, 56.83, 55.93, 50.50, 47.42, 44.73, 38.67, 38.62, 37.78, 36.88, 30.83, 30.66 , 27.82, 27.78, 26.49, 26.42, 24.17, 24.05, 21.23, 21.17, 19.20, 19.16, 16.50, 13.75, 13.56.LC-MS: [M-17] + =427.35

[0559] Example 151

[0560] Preparation of compound 151 cholester-5(6)-ene-3β,4β,25-triol

[0561]

[0562]

[0563] In the first step, (22E,24S)-stigmaster-6(5),22(23)-dien-3β-ol (50.0 g, 0.12 mol, 1.0 eq.) and 4-dimethylaminopyridine (2.96 g, 24.23 mmol) were weighed and dissolved in dichloromethane (500 mL). Triethylamine (61.90 g, 605.77 mmol) was added, followed by acetic anhydride (37.11 g, 0.36 mol) under stirring at room temperature. The mixture was then stirred at room temperature for 2 hours. The reaction was monitored by TLC (PE:EtOAc = 10:1). After the reaction was completed, the reaction solution was concentrated. The crude product was then added to 300 mL of methanol and stirred. After filtration, the filter cake was washed with 20 mL of methanol twice and then dried to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E)-5-ethyl-6-methylhept-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-1) (53.8 g, 112.4 mmol, 92.77%), a white solid. 1 HNMR (399MHz, Chloroform-d) δ5.35 (d, J=4.9Hz, 1H), 5.14 (dd, J=15.1, 8.5Hz, 1H), 4.99 (dd, J=15.2, 8.5Hz, 1H), 4.67-4.48 (m, 1H), 2.30 (d, J=7.8Hz, 2H), 2.01 (s, 3H), 2.00-1. 91 (m, 2H), 1.84 (d, J = 11.3Hz, 2H), 1.68 (dd, J = 9.4, 4.8Hz, 1H), 1.48 (dt, J = 24.5, 9.2Hz, 7H), 1.28-1.07 (m, 6H), 1.00 (d, J=3.2Hz, 6H), 0.80 (dd, J=20.1, 6.4Hz, 9H), 0.68 (s, 3H).

[0564] The second step involves weighing 5.0 g of acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E)-5-ethyl-6-methylhept-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-1). 11 mmol) was dissolved in tetrahydrofuran (200 mL) and water (20.0 mL). Pyridine (2.12 mL, 27.5 mmol), N-methylmorpholine oxide (5.15 g, 43.98 mmol), and potassium osmium tetroxide (7.3 mL, 1.1 mmol) were added at room temperature. The mixture was stirred overnight at room temperature. TLC (PE:EtOAc = 3:1) showed phosphomolybdic acid color development, with some raw material remaining and an intermediate (vicinal diol formed). Sodium periodate (9.41 g, 43.98 mmol) was then added to the reaction mixture at 0 °C. The mixture was stirred at room temperature for 1 hour. TLC (PE:EtOAc = 3:1) showed phosphomolybdic acid color development, with some raw material remaining and the intermediate converted to the product. 50 mL of water and 50 mL of ethyl acetate were added for extraction. The mixture was dried, concentrated, and purified by column chromatography (PE:EtOAc = 60:1) to obtain a white solid. Acid-(1R,3aS,3bS,7S,9aR,9bS,11aS)-1-[(1S)-1-formylethyl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-2) (0.64 g, 1.6 mmol, 14.73%), white solid. 1 HNMR (399MHz, CDCl3) δ9.55 (s, 1H), 5.36 (s, 1H), 4.60 (s, 1H), 2.31 (s, 3H), 2.02 (s, 3H), 1.93 (s, 2H), 1.84 (d, J=11.2Hz, 3H), 1.63 (s, 3H), 1 .49 (d, J=10.9Hz, 4H), 1.42-1.19 (m, 3H), 1.15 (s, 1H), 1.11 (d, J=6.8Hz, 3H), 1.06 (s, 1H), 1.01 (s, 3H), 0.98 (d, J=5.8Hz, 1H), 0.71 (s, 3H).

[0565] The third step involves dissolving (methoxymethyl)triphenylphosphine chloride (8.7 g, 25.0 mmol) in tetrahydrofuran (38 mL), purging with nitrogen three times, cooling to 0 °C, and then adding sodium bis(trimethylsilyl)amino (12.8 mL, 25.0 mmol) dropwise. After the addition is complete, stirring is continued at 0 °C for half an hour. Then, a tetrahydrofuran (18 mL) solution of acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aS)-1-[(1S)-1-formylethyl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-2) (1.9 g, 5.1 mmol) is added. The mixture was stirred at room temperature for 1 hour, and then detected by TLC (PE:EtOAc = 10:1). The reaction of phosphomolybdic acid was observed, indicating that the starting material had reacted completely. Subsequently, 30 mL of water and 15 mL of ethyl acetate were added for extraction. After drying and concentration, the crude product was purified by column chromatography (PE:EtOAc = 100:1 to 30:1) to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2S,3E)-4-methoxybut-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-3) (0.75 g, 1.8 mmol, 34.87%) as a white solid. 1 HNMR (399MHz, CDCl3) δ6.22 (d, J=12.6Hz, 1H), 5.71 (d, J=6.4Hz, 0H), 5.35 (d, J=4.1Hz, 1H), 4.57 (m, 2H), 4.15 (m, 0H), 3.52 (d, J = 10.6Hz, 1H), 3.46 (d, J = 8.7Hz, 3H), 2.59 (d, J = 6.6Hz, 0H ), 2.30 (d, J = 7.6Hz, 3H), 2.01 (s, 3H), 1.95 (d, J = 5.8Hz, 3H), 1.84 (d, J = 10.9Hz, 3H), 1.69 (d, J=4.6Hz, 1H), 1.52(s, 2H), 1.48(s, 2H), 1.14(m, 3H), 1.03(s, 3H), 1.00(s, 3H), 0.67(s, 3H).

[0566] In the fourth step, 0.75 g (1.87 mmol) of acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2S,3E)-4-methoxybut-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-3) was dissolved in tetrahydrofuran (7.5 mL), and 5N hydrochloric acid (2.25 mL, 11.2 mmol) was added. The mixture was then heated to 50 °C and stirred for 1 hour. The mixture was then analyzed by TLC (PE∶EtOAc=10∶1). The reaction was confirmed by colorimetric analysis of phosphomolybdic acid, indicating that the reaction was complete. Then, 15 mL of water and 10 mL of ethyl acetate were added for extraction (3 times). After drying and concentration, the crude product was purified by column chromatography (PE∶EtOAc=50∶1~20∶1) to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-formylpropyl-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-4) (0.52 g, 1.3 mmol, 68.26%), a white solid. 1 HNMR (399MHz, CDCl3) δ9.74 (s, 1H), 5.36 (d, J = 4.4Hz, 1H), 4.60 (s, 1H), 2.44 (d, J = 16.5Hz , 1H), 2.30 (d, J=6.7Hz, 2H), 2.20-2.10 (m, 1H), 2.01 (s, 3H), 1.95 (d, J=14.4Hz, 2H), 1.84 ( d, J=11.5Hz, 3H), 1.57 (dd, J=19.7, 8.6Hz, 4H), 1.46 (dd, J=14.2, 10.3Hz, 3H), 1.29-1.16( m, 3H), 1.15-1.03 (m, 3H), 1.01 (d, J = 4.5Hz, 6H), 0.960.89 (m, 1H), 0.70 (d, J = 12.6Hz, 3H).

[0567] In the fifth step, 0.52 g (1.34 mmol) of acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-formylpropyl-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-4) was dissolved in tetrahydrofuran (30 mL), and 2.25 g (6.73 mmol) of (triphenyl-λ5-methylphosphine)acetic acid methyl ester was added. The reaction system was heated to 90 °C and stirred for 3 hours. HPLC analysis showed no reaction proceeds. The remainder was then cooled to room temperature, and 15 mL of water and 10 mL of ethyl acetate were added for extraction. After drying and concentration, the crude product was purified by column chromatography (PE:EtOAc = 50:1 to 20:1) to obtain methyl (2E,5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (151-5) (0.44 g, 0.94 mmol, 70.20%). 1 HNMR (399MHz, CDCl3) δ6.99-6.86 (m, 1H), 5.80 (d, J=15.6Hz, 1H), 5.36 (s, 1H) , 4.59 (s, 1H), 3.71 (s, 3H), 2.30 (d, J = 6.7Hz, 3H), 2.01 (s, 3H), 2.00-1.90 (m, 3 H), 1.84 (d, J=10.9Hz, 3H), 1.54 (d, J=16.6Hz, 5H), 1.50-1.37 (m, 3H), 1.33-1. 17(m, 2H), 1.16-1.02(m, 4H), 1.00(s, 3H), 0.93(d, J=6.6Hz, 3H), 0.67(s, 3H).

[0568] Step 6: Weigh (2E, 5R)-5-[(1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b, 4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (151-5) (0.44 g, 1.0 mmol) and dissolve it in a mixed solvent of tetrahydrofuran (9.0 mL) and methanol (4.5 mL). Add nickel chloride (130 mg, 1.0 mmol), and then add sodium borohydride (56.92 mg, 1.5 mmol) under stirring at room temperature. The reaction solution contains a large amount of... After bubble generation and stirring at room temperature for 1 hour, a sample was taken, concentrated, and monitored by NMR. The reaction of the raw materials was complete. 15 mL of water and 10 mL of ethyl acetate were added for extraction. After drying and concentration, the crude product was purified by column chromatography (PE:EtOAc = 60:1~20:1) to obtain methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (151-6) (0.36 g, 0.78 mmol, 77.74%), a white solid. 1 HNMR (399MHz, CDCl3) δ5.36 (s, 1H), 4.59 (s, 1H), 3.65 (s, 3H), 2.35-2.18 (m, 4H), 2.01 (s, 3H) , 1.94 (d, J = 15.3Hz, 2H), 1.84 (d, J = 10.9Hz, 3H), 1.68 (s, 1H), 1.55 (s, 2H), 1.53-1.44 (m, 4H) , 1.39 (d, J=10.2Hz, 3H), 1.22 (dd, J=22.8, 12.0Hz, 2H), 1.15-1.09 (m, 2H), 1.08-1.03 (m, 2H) , 1.00 (s, 3H), 0.96 (s, 1H), 0.91 (d, J = 6.3Hz, 3H), 0.89-0.79 (m, 1H), 0.64 (d, J = 12.6Hz, 3H).

[0569] Step 7: Weigh out methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (151-6) (0.15 g, 0.34 mmol) and dissolve it in a chloroform (6.0 mL) mixed solvent. Add... Selenium dioxide (52.4 mg, 0.47 mmol) was added, followed by N-methylmorpholine (102.36 mg, 1.01 mmol) with stirring. The reaction system was heated to 70 °C and refluxed with stirring at this temperature for 64 hours. TLC (PE:EtOAc = 3:1) monitoring showed the formation of highly polar substances. The raw material remained. Subsequently, 10 mL of water was added to quench the reaction, and the mixture was extracted with 10 mL of ethyl acetate three times. After drying and concentration, the crude product was subjected to column chromatography (PE:EtOAc = 1). After purification, the following raw materials were obtained: acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-methoxy-6-oxylidenehex-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (20 mg) and product: (5R)-5-[(1 R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (151-7) (66 mg, 0.14 mmol, 40.35%) was a white solid. Its chiral configuration was confirmed by two-dimensional spectroscopy, and the results are as follows: Figures 54-55 As shown. 1 HNMR (399MHz, CDCl3) δ5.69 (s, 1H), 4.70 (d, J = 11.7Hz, 1H), 4.23 (s, 1H), 3.65 ( s, 3H), 2.25 (dd, J=16.6, 8.5Hz, 2H), 2.09 (s, 3H), 2.05-1.94 (m, 2H), 1.83 (s, 3H ), 1.67(s, 3H), 1.56(s, 3H), 1.45(s, 2H), 1.39(s, 3H), 1.24(s, 2H), 1.20(s, 3H ), 1.17-1.08 (m, 3H), 0.99 (d, J = 9.5Hz, 3H), 0.91 (d, J = 6.4Hz, 3H), 0.65 (s, 3H).

[0570] Step 8: Weigh out methyl (5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (151-7) (30 mg, 0.06 mmol), dissolve it in a mixed solvent of tetrahydrofuran (2.0 mL), purge with nitrogen, and cool to - At 78℃, methyllithium (0.2 mL, 0.33 mmol) was added dropwise, then the mixture was brought to room temperature and stirred for 3 hours. The reaction was monitored by TLC (DCM:MeOH = 20:1) and the reaction of the starting material was complete. Then, 5 mL of water was added to quench the reaction, and 3 mL of ethyl acetate was added for extraction. After drying and concentration, the crude product was purified by column chromatography (DCM:MeOH = 500:1~100:1) to obtain cholester-5(6)-ene-3β,4β,25-triol (151) (13 mg, 0.03 mmol, 45.3%) as a white solid. 1 HNMR (399MHz, Chloroform-d) δ5.71-5.61 (m, 1H), 4.12 (s, 1H), 3.54 (s, 1H), 2.16-1.9 6 (m, 4H), 1.85 (dd, J=28.6, 13.7Hz, 3H), 1.64 (s, 2H), 1.55 (d, J=20.5Hz, 3H), 1.42 (s, 4 H), 1.36 (d, J=11.0Hz, 4H), 1.23 (s, 2H), 1.19 (s, 6H), 1.16 (s, 3H), 1.10 (d, J=10.0Hz, 2 H), 1.05 (d, J=10.5Hz, 2H), 0.99 (d, J=13.0Hz, 2H), 0.91 (d, J=6.5Hz, 3H), 0.66 (s, 3H). 13 CNMR (100MHz, CDCl3) δ142.69, 134.69, 128.79, 77.31, 77.24, 77.00, 76.68, 72.46, 71.11, 56.86, 55.96, 50.12, 44.38, 42.2 9, 39.64, 36.86, 36.40, 35.96, 35.71, 32.05, 31.78, 29.35, 29.18, 28.22, 25.38, 24.22, 21.03, 20.72, 20.50, 18.65, 11.85.

[0571] The two-dimensional spectrum of compound 151 is as follows: Figures 56-57 As shown.

[0572] Example 160

[0573] Preparation of compound 160(1R,3aR,5aR,7S,9aR,11aR)-1-[(2R)-6-hydroxyoct-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol

[0574]

[0575] (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-6-hydroxyoctyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (102) (200 mg, 0.38 mmol) was dissolved in chloroform. Hydrogen chloride gas (prepared on-site: concentrated sulfuric acid was added dropwise to solid sodium chloride at a gas flow rate of approximately 5 bubbles / min) was introduced. After 3 hours of purging, the reaction was stopped, and the reaction solution was treated with saturated sodium bicarbonate. Washed with saturated brine, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (PE:EtOAc = 90:10) to obtain crude product with a purity of 40%. The crude product was further purified by chirality to obtain a white solid (1R,3aR,5aR,7S,9aR,11aR)-1-[(2R)-6-hydroxyoct-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (160) (22 mg, 0.05 mmol, 27%). 1 HNMR (400MHz, CDCl3) δ5.24-5.18 (m, 1H), 3.56-3.48 (m, 1H), 3.25 (dd, J=10.9, 4.7Hz, 1H), 2.06-1.88 (m, 4H), 1.83-1.76 (m, 1H) , 1.61 (m, 5H), 1.51-1.33 (m, 10H), 1.29-1.03 (m, 6H), 0.99 (s, 3H), 0.98-0.92 (m, 6H), 0.89 (dd, J=7.6, 4.7Hz, 9H), 0.64 (s, 3H). 13CNMR (101MHz, CDCl3) δ144.93, 116.66, 79.35, 77.34, 77.02, 76.70, 51.98, 50.72, 50.19, 47.22, 44.36, 38.63, 38.11, 37.52, 37.39, 36.43, 3 6.38, 36.35, 36.28, 35.60, 32.16, 32.08, 28.24, 27.63, 27.49, 24.83, 22.95, 22.35, 19.98, 18.94, 16.07, 16.04, 15.48, 14.13.LC-MS: [M-17] + =427.4

[0576] Example 172

[0577] Synthesis of compound 172(1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-6-hydroxyoct-2-yl]-3a,4,6,6,9a,11a-hexamethyl-2,3,3a,5a,6,7,8,9,9a,10,11,11a-dodecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol

[0578]

[0579] Step 1: (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-6-hydroxyoct-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (102) (2 g, 4.50 mmol, 1.0 eq) was dissolved in dichloromethane (100 mL), followed by the addition of acetic anhydride (2.1 mL, 22.49 mmol, 5.0 eq), DMAP (0.11 g, 0.90 mmol, 0.2 eq), and triethylamine (3.13 mL, 22.49 mmol, 5.0 eq). The mixture was stirred at room temperature for 2 hours, and the reaction was monitored at TC (petroleum ether:ethyl acetate = 10:1). After the reaction was completed, the reaction solution was quenched with methanol, washed with saturated sodium bicarbonate and saturated brine respectively, dried over anhydrous sodium sulfate, purified by silica gel column chromatography, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain a white waxy substance, acetic acid-(7R)-7-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]octyl-3-yl ester (172-1) (2.5 g, 4.02 mmol, 89.4%). 1 H NMR (400MHz, CDCl3) δ4.82 (d, J=5.5Hz, 1H), 4.50 (dd, J=11.5, 4.5Hz, 1H), 2.08-1.97 (m, 10H), 1.94-1.85 (m, 1H), 1.78-1 .59 (m, 7H), 1.55-1.23 (m, 11H), 1.16 (dd, J=17.5, 8.2Hz, 3H), 1.0l (d, J=7.9Hz, 4H), 0.88 (t, J=6.6Hz, 15H), 0.68 (s, 3H).

[0580] In the second step, acetic acid-(7R)-7-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]octyl-3-yl ester (172-1) (2.5 g, 4.73 mmol, 1.0 eq) was dissolved in dichloromethane (100 mL). Under nitrogen protection and an ice-water bath, m-CPBA (0.82 g, 4.73 mmol, 1.0 eq) was added in portions, followed by reaction at room temperature. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was complete, the phase was quenched with saturated sodium bisulfite, washed with saturated sodium bicarbonate, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 95:5 to 85:15) to give a white waxy compound 2-[(2R,5R,6R,11S)-5-[(2R)-6-acetoxyoctyl-2-yl]-2,6,10,10-tetramethyl-14-oxatetracyclo[7.4.1.01,9.02,6]tetradecane-11-yl]-2-methylpropyl ester (172-2) (2 g, 2.94 mmol, 62.1%). 1 H NMR (400MHz, CDCl3) δ4.81 (s, 1H), 4.46 (s, 1H), 2.04 (d, J = 6.9Hz, 7H), 1.99-1.81 (m, 5H), 1.76-1.67 (m, 4H), 1. 66-1.45 (m, 11H), 1.34 (m, 11H), 0.97 (d, J=9.5Hz, 2H), 0.87 (dd, J=11.7, 7.0Hz, 13H), 0.82 (s, 3H), 0.76 (s, 3H).

[0581] In the third step, acetic acid-2-[(2R,5R,6R,11S)-5-[(2R)-6-acetoxyoctyl-2-yl]-2,6,10,10-tetramethyl-14-oxatetracyclo[7.4.1.01,9.02,6]tetradecane-11-yl]-2-methylpropyl ester (172-2) (1.9 g, 3.49 mmol, 1.0 eq) was dissolved in acetone (100 mL), and 10 drops of concentrated sulfuric acid were added. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was complete, acetone was evaporated by rotary evaporation, 100 mL of ethyl acetate was added, and the mixture was washed with saturated bicarbonate, then with saturated brine, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 90:10 to 80:20) to give a white solid acetic acid-(7R)-7-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,5,5a,6,7,8,9,9a,11,11a-dodecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]octyl-3-yl ester (172-3) (1 g, 1.71 mmol, 49.0%). 1 H NMR (400MHz, CDCl3) δ5.53 (s, 1H), 5.39 (d, J = 6.0Hz, 1H), 4.89 (t, J = 6.1Hz, 1H), 4.59 (dd, J = 11.1, 4.8Hz, 1H), 2.20-2.10 (m, 9H), 2.10-2.03 (m, 2H), 1 .81(ddd, J=11.7, 7.5, 3.5Hz, 2H), 1.75-1.53(m, 9H), 1.49-1.32(m, 8H), 1. 29-1.22(m, 2H), 1.08(s, 3H), 1.03(s, 3H), 0.980.93(m, 15H), 0.63(s, 3H). 13C NMR (101MHz, CDCl3) δ170.99, 170.96, 145.60, 142.78, 119.87, 116.58, 80.83, 77.33, 77.01, 7 6.70, 75.58, 75.51, 50.99, 50.32, 49.25, 43.72, 37.82, 37.61, 37.23, 36.20, 36.17, 36.04, 36 .00, 35.40, 34.03, 31.88, 31.59, 31.48, 28.09, 27.91, 27.07, 26.91, 26.88, 25.52, 24.26, 22.82, 22.78, 22.66, 22.19, 22.14, 21.33, 21.28, 18.42, 18.40, 16.93, 15.65, 14.12, 9.62, 9.59.

[0582] In the fourth step, acetic acid-(7R)-7-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,5,5a,6,7,8,9,9a,11,11a-dodecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]octyl-3-yl ester (172-3) (0.9 g, 1.71 mmol, 1.0 eq) was dissolved in glacial acetic acid (30 mL). Under nitrogen protection and in an ice-water bath, 30% hydrogen peroxide (1.7 mL, 17.1 mmol, 10.0 eq) was added dropwise. After stirring for 10 minutes, the reaction was brought to room temperature and stirred. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1, phosphomolybdic acid plate). After the reaction was completed, the product was diluted with ethyl acetate, washed twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain crude acetic acid-(7R)-7-[(1R,3aS,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-4-oxonyl-2,3,3a,3b,4,5,5a,6,7,8,9,9a,11,11a-tetradecanoyl-1H-cyclopenta[1,2-a]phenanthrene-1-yl]octyl-3-yl ester (172-4) (350 mg, 0.52 mmol, 30.2%). The crude product was directly fed into the next step.

[0583] Step 5: Acetic acid-(7R)-7-[(1R,3aS,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-4-oxoylide-2,3,3a,3b,4,5,5a,6,7,8,9,9a,11,11a-tetradecanoyl-1H-cyclopenta[1,2-a]phenanthrene-1-yl]octyl-3-yl ester (172-4) (200 mg, 0.37 mmol) was dissolved in a mixed solvent of chloroform (3 mL) and methanol (1.5 mL). Under nitrogen protection, concentrated hydrochloric acid (36%-38%) (0.18 mL) was added dropwise in an ice-water bath. After stirring for 10 minutes, the mixture was allowed to cool to room temperature. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1, phosphomolybdic acid plate). After the reaction was completed, the reaction mixture was diluted with water, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 80:20) to give acetate-(7R)-7-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-4-oxoylide-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]octyl-3-yl ester (172-5) (100 mg, 0.12 mmol, 32.5%). 1 H NMR (400MHz, CDCl3) δ4.85-4.76 (m, 1H), 4.51 (dd, J=11.6, 4.3Hz, 1H), 2.42 (t , J=8.6Hz, 1H), 2.29 (dt, J=17.0, 6.9Hz, 1H), 2.06 (d, J=5.6Hz, 6H), 2.00-1.90 (m, 1H), 1.84-1.69 (m, 5H), 1.60-1.46 (m, 5H), 1.41-1.20 (m, 8H), 1.19-1.13 (m , 3H), 1.02 (m, 2H), 0.96 (d, J=8.2Hz, 3H), 0.86 (m, 14H), 0.63 (d, J=8.5Hz, 2H). 13C NMR (101MHz, CDCl3) δ198.73, 171.01, 170.83, 79.62, 77.34, 77.02, 76.70, 75.53, 49.86, 49.03, 47.79, 44.90, 39.62, 37.76, 36.45, 36. 27, 34.50, 34.05, 31.99, 30.09, 27.38, 27.10, 26.91, 25.02, 23.85, 23.70, 22.12, 21.29, 21.24, 18.72, 18.44, 16.37, 15.79, 9.62, 9.59.

[0584] Step 6: Acetic acid-(7R)-7-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetoxy-3a,6,6,9a,11a-pentamethyl-4-oxonyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]octyl-3-yl ester (172-5) (100 mg, 0.18 mmol, 1.0 eq) was dissolved in anhydrous tetrahydrofuran (1 mL) under nitrogen protection. Methyl magnesium chloride solution (1.0 mL, 3.0 mol / L, 20.0 eq) was added dropwise at 0 °C, and then the reaction was allowed to proceed to room temperature. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was completed, the mixture was quenched with saturated ammonium chloride solution in an ice-water bath, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 90:10) to give a white solid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-6-hydroxyoctyl-2-yl]-3a,4,6,6,9a,)1a-hexamethyl-2,3,3a,5a,6,7,8,9,9a,10,11,11a-dodecylhydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol (172) (5.88 mg, 0.013 mmol, 7.2%). 1HNMR (400MHz, CDCl3) δ5.25 (d, J=5.6Hz, 1H), 3.57-3.49 (m, 1H), 3.23 (dd, J=11.4 , 4.3Hz, 1H), 2.26-2.09(m, 3H), 2.09-1.94(m, 2H), 1.94-1.87(m, 2H), 1.87(s, 3H), 1.75-1.60 (m, 3H), 1.56-1.46 (m, 3H), 1.41-1.15 (m, 8H), 1.09 (dd, J=12.4, 3.9Hz, 2H), 1.00 (s, 3H), 0.94 (dd, J=13.6, 5.4Hz, 9H), 0.88 (d, J=6.0Hz, 6H), 0.64 (s, 3H).13 C NMR(101 MHz, CDCl3) δ147.04, 135.31, 129.36, 114.44, 78.96, 73.46, 73.36, 52.14, 49.51, 48.02, 45.05, 38.51, 37.53, 37.42, 37.29, 36.53, 36.33, 36 .24, 36.17, 35.82, 33.04, 30.26, 30.13, 28.58, 27.96, 27.85, 23.36, 22 .45, 22.36, 22.23, 22.08, 18.75, 16.27, 15.41, 9.90, 9.84.LCMS[M-OH] + =439.7

[0585] Example 184

[0586] Preparation of compound 184(23E)-cholest-5(6),23(24)-diene-3β,25-diol

[0587]

[0588]

[0589] Acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,4E)-6-methoxy-6-oxoylide-4-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-5) (80 mg, 0.181 mmol) was dissolved in tetrahydrofuran (3 mL), cooled to -78 °C, and methyllithium (0.565 mL, 0.904 mmol) was added. The mixture was then heated to room temperature and stirred for three hours. TLC (petroleum ether:ethyl acetate = 10:1) was used for monitoring. Water (10 mL) and ethyl acetate (5 mL * 3) were added to the reaction solution for extraction. After combining the organic phases, the mixture was washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain 90 mg of crude product. The crude product was purified by column chromatography (dichloromethane:methanol = 1:0-100:1) to obtain a white solid (23E)-cholest-5(6),23(24)-diene-3β,25-diol (184) (50 mg, 0.119 mmol, 65.60%). 1HNMR (399MHz, CDCl3) δ5.56 (s, 2H), 5.34 (s, 1H), 3.51 (s, 1H), 2.25 (dd, J=18 .7, 12.7Hz, 2H), 2.16-2.09 (m, 1H), 1.97 (d, J=12.2Hz, 2H), 1.82 (d, J=9.7Hz, 3H), 1.74 (d, J=14.0Hz, 1H), 1.58 (s, 3H), 1.47 (s, 4H), 1.44-1.37 (m, 3H), 1.2 9(s, 6H), 1.18-1.04(m, 4H), 0.98(s, 3H), 0.88(d, J=6.6Hz, 3H), 0.66(s, 3H). 13 CNMR (100MHz, CDCl3) δ140.72, 139.34, 125.44, 121.64, 77.31, 77.19, 76.99, 76.67, 71.75, 70.73, 56.66, 55.74, 50.04, 42.32, 42. 25, 39.63, 38.78, 37.20, 36.46, 36.06, 31.85, 31.61, 30.93, 29.88, 29.87, 28.18, 24.28, 21.02, 19.37, 18.63, 11.88.LCMS: [M-OH] + =383.30

[0590] Example 187

[0591] Preparation of compound 1872-fluoro-N-[(3R)-3-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]butyl]-N-methylbenzamide

[0592]

[0593]

[0594] In the first step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-formylpropyl-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-4) (100 mg, 0.259 mmol) was dissolved in THF, and methylamine (32.18 mg, 1.036 mmol) and zinc chloride (17.65 mg, 0.130 mmol) were added. After stirring for 1 hour, sodium cyanoborohydride (81.38 mg, 1.295 mmol) was added, and the reaction system was stirred overnight at room temperature. TLC monitoring (petroleum ether:ethyl acetate = 10:1) showed no remaining starting material. Water (10 mL) and ethyl acetate (8 mL * 3) were added to the reaction solution for extraction. The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-9a,11a-dimethyl-1-[(2R)-4-(methylamino)but-2-yl]-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (187-1) (120 mg, 0.209 mmol, 80.85%), which was directly proceeded to the next step without purification.

[0595] In the second step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-9a,11a-dimethyl-1-[(2R)-4-(methylamino)but-2-yl]-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (187-1) (120 mg, 0.209 mmol) and triethylamine (0.087 mL, 0.627 mmol) were dissolved in dichloromethane (3 mL), and o-fluorobenzoyl chloride (66.32 mg, 0.418 mmol) was added. The mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 3:1) and LC-MS. Add water (15 mL) and ethyl acetate (10 mL * 3) to the reaction solution for extraction. Wash the organic phase once with saturated brine, dry with anhydrous sodium sulfate, and concentrate to obtain 60 mg of crude product (187-2). Proceed directly to the next step.

[0596] In the third step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-4-{[(2-fluorophenyl)carbonyl](methyl)amino}but-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (187-2) (60 mg, 0.115 mmol) was dissolved in tetrahydrofuran (2 mL) and methanol (1 mL), and sodium hydroxide aqueous solution (1 mL, 1.000 mmol) was added. The mixture was stirred at room temperature for 8 hours. TLC (petroleum ether:ethyl acetate = 3:1) monitoring. Water (10 mL) and ethyl acetate (8 mL * 3) were added to the reaction solution and extracted three times. The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain 60 mg of crude product. The crude product was purified by preparative liquid chromatography to obtain 2-fluoro-N-[(3R)-3-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]butyl]-N-methylbenzamide (187) (30 mg, 0.059 mmol, 51.64%), a white solid. 1 H NMR (399MHz, CDCl3) δ7.39-7.28 (m, 2H), 7.17 (t, J=7.4Hz, 1H), 7.07 (q, J=8.4Hz, 1H), 5.33 (s, 1H), 3.68 (s, 0.5H), 3.55-3.47 (m, 1H), 3.46-3.37 (m, 0.5H), 3.17 (d, J=10.5Hz, 1H), 3.07 (s, 1.5H), 2.86 (s, 1.5H), 2.23 (d, J=20.6Hz, 2H), 1.94 (d, J=40.2Hz, 3H), 1.82 (t, J= 9.7Hz, 3H), 1.63 (s, 2H), 1.53-1.45 (m, 4H), 1.40 (d, J = 11.5Hz, 2H), 1.32 (s, 2H), 1.15 (d, J = 9.3Hz, 2H), 1.08 (d, J = 9.7Hz, 2H), 1.03 (d , J=6.5Hz, 1.5H), 0.99 (s, 1.5H), 0.97 (s, 1.5H), 0.91 (dd, J=17.3, 7.5Hz, 2H), 0.69 (s, 1.5H), 0.63 (d, J=5.8Hz, 1.5H), 0.59 (s, 1.5H). 13C NMR (101MHz, CDCl3) δ140.71, 121.66, 121.57, 77.32, 77.20, 77.00, 76.68 ,71.75,56.72,56.61,55.88,55.52,50.04,49.97,44.89,42.23,39.72,3 9.58, 37.19, 36.44, 33.77, 33.42, 32.74, 31.87, 31.80, 31.63, 30.95, 28. 29, 28.06, 24.26, 24.16, 21.05, 20.97, 19.39, 18.79, 18.49, 11.78, 11.70. 19 FNMR (376MHz, CDCl3) δ-115.21, -115.37.LC-MS: [M+H] + =482.50

[0597] Example 195

[0598] Preparation of compound 195(3S,8S,9S,10R,13R,14S,17R,20R,22E)-N-(2-fluorophenyl)-3-hydroxy-N-methylcholan-5(6),22(23)-diene-24-amide

[0599]

[0600] The first step involved weighing acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aS)-1-[(1S)-1-formylethyl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-2) (400 mg, 1.1 mol, 1.0 eq.) and dissolving it in tetraoxofuran (24 mL). Then, methyl (triphenyl-λ5-methylphosphine)acetate (1.8 g, 5.37 mmol) was added, and the system was heated to 90 °C and stirred for 3 hours. The reaction solution was concentrated, and water (30 mL) and ethyl acetate (10 mL * 3) were added to the reaction solution for extraction. The mixture was washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain 1.9 g of crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 0%–1%) to give acetic acid-(1R,3aS,3bS,7S,9aR,9bS,)1aR)-1-[(2R,3E)-5-methoxy-5-oxoylidene-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (195-1) (350 mg, 0.776 mmol, 72.25%) as a white solid. 1 H NMR (399MHz, CDCl3) δ6.82 (dd, J=15.6, 9.0Hz, 1H), 5.72 (d, J=15.6Hz, 1H), 5.35 (s, 1H), 4.57 (s, 1H), 3.70 (s, 3H), 2.34-2.16 (m, 4H), 2.02 (s, 3H), 1.99-1.90 (m, 3H), 1.84 (d, J= 11.4Hz, 2H), 1.65 (dd, J=19.5, 8.0Hz, 2H), 1.49 (s, 2H), 1.46-1.41 (m, 2H), 1.23 (d, J=11 .7Hz, 3H), 1.13 (d, J=10.7Hz, 3H), 1.07 (d, J=6.6Hz, 3H), 1.00 (s, 3H), 0.72-0.65 (m, 3H).

[0601] In the second step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E)-5-methoxy-5-oxoylidene-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (195-1) (380 mg, 0.88 mmol) was dissolved in tetrahydrofuran (10 mL) and methanol (5 mL). NaOH (3 mL, 3.000 mmol) was added to the above solution, and the mixture was stirred overnight at room temperature. TLC monitoring (DCM∶MeOH=10∶1) showed that the reaction of the raw materials was complete. The reaction solution was concentrated to obtain 280 mg of solid. THF (10 mL) was added and the mixture was stirred for 2 hours. After filtration, the filtrate was concentrated to obtain (22E)-3β-hydroxycholan-5(6),22(23)-diene-24-acid (195-2) (319 mg, 0.813 mmol, 91.75%) white solid.

[0602] In the third step, (22E)-3β-hydroxycholan-5(6),22(23)-diene-24-acid (195-2) (319 mg, 0.813 mmol) was weighed and dissolved in tetrahydrofuran (10 mL). Triethylamine (0.357 mL, 2.569 mmol) and 4-dimethylaminopyridine (10.46 mg, 0.086 mmol) were added, followed by acetic anhydride (0.161 mL, 1.713 mmol). The mixture was then kept at room temperature. The mixture was stirred for 3 hours and monitored by TLC (DCM:MeOH = 10:1). The reaction of the starting material was complete. Then, 15 mL of water was added, and the mixture was extracted with ethyl acetate (10 mE*). After drying and concentration, the crude product was purified by column chromatography (DCM:MeOH = 0%~1%) to obtain (22E)-3β-acetoxycholan-5(6),22(23)-diene-24-acid (195-3) (137 mg, 0.314 mmol, 36.66%) as a white solid. 1 H NMR (399MHz, Chloroform-d) δ5.35 (d, J=5.5Hz, 1H), 4.58 (d, J=9.8Hz, 1H), 2.4 4-2.20(m, 4H), 2.01(s, 3H), 1.95(t, J=12.2Hz, 2H), 1.88-1.76(m, 4H), 1.56(q , J=11.4, 10.7Hz, 3H), 1.49-1.39 (m, 4H), 1.38-1.25 (m, 3H), 1.17-1.03 (m, 4H) , 0.99 (s, 3H), 0.91 (d, J = 6.5Hz, 3H), 0.85 (dd, J = 18.9, 11.7Hz, 1H), 0.65 (s, 3H)

[0603] In the fourth step, (22E)-3β-acetoxycholan-5(6),22(23)-diene-24-acid (110 mg, 0.26 mmol) (195-3) was weighed and dissolved in dichloromethane (10 mL). N,N'-dimethylformamide (0.001 mL, 0.014 mmol) and oxaloyl chloride (0.11 mL, 1.33 mmol) were added. After stirring at room temperature for 1 hour, the mixture was concentrated to obtain a solid. The solid was dissolved in dichloromethane (2 mL) and added to a dichloromethane (3 mL) solution of o-fluoroaniline (58.97 mg, 0.53 mmol) and triethylamine (0.11 mL, 0.80 mmol). The mixture was stirred at room temperature for 1 hour and monitored by TLC (petroleum ether: ethyl acetate = 5:1). After the reaction was completed, 15 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL * 3). After drying and concentration, the crude product was purified by preparative TLC to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E)-5-[(2-fluorophenyl)amino]-5-oxoylidenepent-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (195-4) (80 mg, 0.145 mmol, 54.64%) as a white solid.

[0604] In the fifth step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E)-5-[(2-fluorophenyl)amino]-5-oxoylidenepent-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (195-4) (70 mg, 0.14 mmol) was dissolved in N,N'-dimethylformamide (5 mL), purged with nitrogen three times, sodium hydride (16.55 mg, 0.69 mmol) was added, and after stirring for half an hour, a solution of iodomethane (29.36 mg, 0.21 mmol) in N,N'-dimethylformamide (2 mL) was added, and the mixture was stirred at room temperature for 1 hour. TLC (petroleum ether: ethyl acetate = 5:1) monitoring was performed. After the reaction was completed, 15 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL * 3). After drying and concentration, the crude product, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E)-5-[(2-fluorophenyl)(methyl)amino]-5-oxoylidenepent-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (195-5) (80 mg), was directly added to the next reaction without further purification.

[0605] In step six, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E)-5-[(2-fluorophenyl)(methyl)amino]-5-oxoylidenepent-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (195-5) (40 mg, 0.077 mmol) was dissolved in tetrahydrofuran (4 mL) and methanol (2 mL), and sodium hydroxide (1 mL, 1.000 mmol) was added. The mixture was stirred at room temperature for 16 h. TLC (petroleum ether: ethyl acetate = 5:1) detection. After the reaction was completed, 10 mL of water was added to quench the reaction. Ethyl acetate (10 mL * 30) was extracted, dried, concentrated and the crude product was purified by HPLC to obtain (3S, 8S, 9S, 10R, 13R, 14S, 17R, 20R, 22E)N-(2-fluorophenyl)-3-hydroxy-N-methylcholan-5(6),22(23)-diene-24-amide (195) (20 mg, 0.040 mmol, 51.66%) white solid. 1H NMR (399MHz, CDCl3) δ7.32 (d, J=7.6Hz, 1H), 7.20-7.10 (m, 3H), 6.73 (dd, J=14.8, 9.2Hz, 1H), 5. 55 (d, J=15.3Hz, 1H), 5.32 (d, J=5.0Hz, 1H), 3.50 (s, 1H), 3.26 (s, 3H), 2.31-2.18 (m, 2H), 2.11-1 .85 (m, 4H), 1.81 (d, J = 10.2Hz, 2H), 1.61 (s, 1H), 1.51 (s, 2H), 1.40 (d, J = 16.4Hz, 2H), 1.17 (d, J = 8.7Hz, 3H), 1.03 (d, J=12.2Hz, 3H), 0.96 (d, J=10.2Hz, 6H), 0.86 (s, 2H), 0.59 (d, J=12.5Hz, 3H). 13 C NMR (100MHz, CDCl3) δ166.75, 152.11, 140.67, 129.88, 129.37, 124.75, 121.58, 118.16, 116.82, 116.71, 77.30, 77.19, 76.99, 76.67, 71.71, 56.46, 54.83, 50.00, 42.50, 42.22, 39.86, 39.50, 37.19, 36.44, 31.83 , 31.77, 31.59, 30.93, 24.26, 20.98, 19.41, 19.36, 12.02.LC-MS: [M+H] + =480.35.

[0606] Example 196

[0607] Preparation of compound 196(3S,8S,9S,10R,13R,14S,17R,20R)-N-(2-fluorophenyl)-3-hydroxy-N-methylcholan-5(6)-ene-24-amide

[0608]

[0609]

[0610] In the first step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E)-5-methoxy-5-oxomylidene-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (195-1) (356 mg, 0.831 mmol) and nickel chloride (107.64 mg, 0.83 mmol) were dissolved in tetrahydrofuran (7 mL) and methanol (3.5 mL). Then, sodium borohydride (56.92 mg, 1.5 mmol) was added under stirring at room temperature. The reaction solution produced a large amount of gas. After foaming and stirring at room temperature for 1 hour, a sample was taken, concentrated, and monitored by NMR. The reaction of the raw materials was complete. 15 mL of water and 10 mL of ethyl acetate were added for extraction. After drying and concentration, the crude product was purified by column chromatography (petroleum ether:ethyl acetate = 60:1 to 20:1) to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5-methoxy-5-oxonylpentan-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (196-1) (270 mg, 0.596 mmol, 71.71) as a white solid. 1 H NMR (399MHz, CDCl3) δ5.35 (d, J=4.6Hz, 1H), 4.58 (d, J=9.6Hz, 1H), 3.64 (s, 3H), 2.3 8-2.15(m, 4H), 2.01(d, J=5.4Hz, 3H), 1.99-1.90(m, 2H), 1.88-1.74(m, 4H), 1.59(d, J=12.6Hz, 2H), 1.49 (dd, J=30.3, 12.1Hz, 4H), 1.42-1.29 (m, 3H), 1.29-1.16 (m, 2H), 1.08 (ddd, J=21.5, 15.4, 7.4Hz, 4H), 0.99 (s, 3H), 0.90 (d, J=6.4Hz, 3H), 0.65 (s, 3H)

[0611] In the second step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5-methoxy-5-oxonylpentan-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (196-1) (270 mg, 0.627 mmol) was dissolved in tetrahydrofuran (10 mL) and methanol (5 mL). NaOH (3 mL, 3.000 mmol) was added to the above solution and stirred overnight at room temperature. TLC monitoring (DCM∶MeOH=10∶1) showed that the raw materials reacted completely. The pH was adjusted to 6-7, and the reaction solution was concentrated to obtain 280 mg of solid. THF (10 mL) was added and the mixture was stirred for 2 hours. After filtration, the filtrate was concentrated to obtain 3β-hydroxycholan-5(6)-en-24-acid (196-2) (215 mg, 0.545 mmol, 86.97%) as a white solid. 1 H NMR (400MHz, cd3od) δ5.33 (d, J=5.4Hz, 1H), 3.43-3.33 (m, 1H), 2.32 (ddd, J=15. 1, 9.7, 5.3Hz, 1H), 2.20 (d, J=9.5Hz, 3H), 2.07-1.93 (m, 2H), 1.90-1.73 (m, 4H), 1.57 (d, J=26.6Hz, 3H), 1.51-1.42 (m, 4H), 1.36-1.25 (m, 3H), 1.17 (dd, J=17.1, 7.1Hz, 2H), 1.12-1.03 (m, 3H), 1.01 (s, 3H), 0.95 (d, J=6.5Hz, 3H), 0.72 (s, 3H).

[0612] In the third step, 3β-hydroxycholan-5(6)-en-24-acid (196-2) (80 mg, 0.214 mmol) was weighed and dissolved in tetrahydrofuran (3 mL). Triethylamine (0.089 mL, 0.642 mmol) and 4-dimethylaminopyridine (2.61 mg, 0.021 mmol) were added, followed by acetic anhydride (0.040 mL, 0.427 mmol). The mixture was stirred at room temperature for 3 hours. TLC (DCM:MeOH = 10:1) was used for detection. Phosphomolybdic acid was observed, indicating that the reaction of the starting material was complete. Then, 15 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL * 3). After drying and concentration, the crude product was purified by column chromatography (DCM:MeOH = 0% ~ 1%) to obtain 3β-acetoxycholan-5(6)-en-24-acid (196-3) (68 mg, 0.155 mmol, 72.46%) as a white solid. 1H NMR (399MHz, Chloroform-d) δ5.35 (d, J=5.5Hz, 1H), 4.58 (d, J=9.8Hz, 1H), 2.4 4-2.20(m, 4H), 2.01(s, 3H), 1.95(t, J=12.2Hz, 2H), 1.88-1.76(m, 4H), 1.56(q, J=11.4, 10.7Hz, 3H), 1.49-1.39(m, 4H), 1.38-1.25(m, 3H), 1.17-1.03(m, 4H), 0.99 (s, 3H), 0.91 (d, J = 6.5Hz, 3H), 0.85 (dd, J = 18.9, 11.7Hz, 1H), 0.65 (s, 3H).

[0613] In the fourth step, 60 mg (0.144 mmol) of 3β-acetoxycholan-5(6)-en-24-acid (196-3) was weighed and dissolved in 3 mL of dichloromethane. N,N'-dimethylformamide (0.001 mL, 0.014 mmol) and oxaloyl chloride (0.061 mL, 0.720 mmol) were added. After stirring at room temperature for 1 hour, the mixture was concentrated to obtain a solid. The solid was dissolved in 2 mL of dichloromethane and added to a 3 mL solution of o-fluoroaniline (32.01 mg, 0.288 mmol) and triethylamine (0.060 mL, 0.432 mmol). The mixture was stirred at room temperature for 1 hour and monitored by TLC (petroleum ether: ethyl acetate = 5:1). After the reaction was completed, 15 mL of water and 10 mL of ethyl acetate were added for extraction. The mixture was dried, concentrated, and the crude product was purified by preparative TLC to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5-[(2-fluorophenyl)amino]-5-oxoylidenepent-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (196-4) (30 mg, 0.056 mmol, 38.82%) as a white solid.

[0614] In the fifth step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5-[(2-fluorophenyl)amino]-5-oxoylidenepent-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (196-4) (50 mg, 0.098 mmol) was dissolved in N,N'-dimethylformamide (3 mL), purged with nitrogen three times, sodium hydride (11.77 mg, 0.490 mmol) was added, and after stirring for half an hour, iodomethane (20.89 mg, 0.147 mmol) in N,N'-dimethylformamide (2 mL) solution was added, and the mixture was stirred at room temperature for 1 hour. TLC (petroleum ether: ethyl acetate = 5:1) monitoring, after the reaction was completed, 15 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL * 3), dried, and concentrated. The crude product, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5-[(2-fluorophenyl)(methyl)amino]-5-oxoylidenepent-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (196)-5 (80 mg), was directly added to the next reaction without further purification.

[0615] In step six, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5-[(2-fluorophenyl)(methyl)amino]-5-oxoylidenepent-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (80 mg, 0.076 mmol) was dissolved in tetrahydrofuran (4 mL) and methanol (2 mL), and sodium hydroxide (1 mL, 1.000 mmol) was added. The mixture was stirred at room temperature for 16 h. TLC (PE∶EtOAc=5∶1) detection. After the reaction was completed, 10 mL of water was added to quench the reaction. Ethyl acetate was extracted with 10 mL*3. After drying and concentration, the crude product was purified by preparative HPLC to obtain (3S,8S,9S,10R,13R,14S,17R,20R)-N-(2-fluorophenyl)-3-hydroxy-N-methylcholan-5(6)-en-24-amide (196) (20 mg, 0.039 mmol, 51.90%) white solid. 1H NMR (399MHz, CDCl3) δ7.33 (d, J=7.9Hz, 1H), 7.22-7.14 (m, 3H), 5.32 (d, J=5.2Hz, 1H), 3.50 (s, 1H), 3.20 (s, 3H), 2.31-2.18 (m, 2H), 2.08 (s, 1H), 1.91 (t, J=13.9Hz, 3H), 1.80 (d, J=9.3 Hz, 2H), 1.76-1.67 (m, 2H), 1.50 (d, J=14.6Hz, 3H), 1.46-1.41 (m, 3H), 1.18 (s, 2H), 1.08-0 .99 (m, 3H), 0.96 (s, 3H), 0.88 (dd, J=16.1, 11.3Hz, 3H), 0.69 (d, J=6.0Hz, 3H), 0.59 (s, 3H). 13 C NMR (101MHz, CDCl3) δ173.95, 145.18, 140.68, 129.70, 125.02, 121.67, 117.01, 116.81, 112.93, 77.31, 77.20, 77.00, 76.68, 71.75, 5 6.61, 56.28, 50.01, 42.25, 39.63, 37.20, 36.44, 35.42, 31.82, 31.60, 30.94, 27.96, 24.22, 21.01, 19.36, 18.21, 11.80.LC-MS: [M+H] + =482.35

[0616] Example 197

[0617] Preparation of compound 1972-[(3R)-3-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopentano[1,2-a]phenanthrene-1-yl]butyl]-1,2,3,4-tetrahydroisoquinoline-1-one

[0618]

[0619] In the first step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-formylpropyl-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-4) (100 mg, 0.26 mmol) was dissolved in tetrahydrofuran (6.0 mL) and methanol (3.0 mL), cooled in an ice bath, and sodium borohydride (19.7 mg, 0.52 mmol) was added at 0 °C. The mixture was then brought to room temperature and stirred at room temperature for 1 hour. The mixture was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, 15 mL of water and ethyl acetate (10 mL * 3) were added for extraction. The organic phase was separated, dried, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 30:1 to 10:1) to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-4-hydroxybutyl-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (197-1) (100 mg, 0.26 mmol, 99% yield) as a white solid.

[0620] 1 H NMR (399MHz, Chloroform-d) δ5.35 (d, J=5.2Hz, 1H), 4.63-4.53 (m, 1H), 3.74-3.5 6(m, 2H), 2.34-2.25(m, 2H), 2.01(s, 3H), 1.95(d, J=18.4Hz, 2H), 1.86-1.82(m, 2H ), 1.72 (ddt, J=10.6, 8.0, 3.9Hz, 1H), 1.62-1.51 (m, 4H), 1.49-1.40 (m, 4H), 1.31 -1.18(m, 5H), 1.16-1.06(m, 4H), 1.00(s, 3H), 0.94(d, J=6.6Hz, 3H), 0.67(s, 3H).

[0621] In the second step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-4-hydroxybutyl-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (197-1) (100 mg, 0.26 mmol) was dissolved in dichloromethane (6.0 mL), and triethylamine (0.09 mL) and 4-dimethylaminopyridine (0.24 mg, 0.002 mmol) were added. Then, p-toluenesulfonyl chloride (148.7 mg, 0.78 mmol) was added while stirring at room temperature. The mixture was stirred at room temperature for 16 hours and monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, 15 mL of water and ethyl acetate (10 mL * 3) were added for extraction. The organic phase was separated, dried, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 30:1 ~ 10:1) to obtain 4-methylbenzenesulfonic acid-(3R)-3-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-a]phenanthrene-1-yl]butyl ester (197-2) (60 mg, 0.11 mmol, 42.31% yield) white solid.

[0622] 1 H NMR (399MHz, Chloroform-d) δ7.76 (d, J=8.1Hz, 2H), 7.32 (d, J=8.0Hz, 2H), 5.33 (d, J=5.0Hz, 1H), 4 .58 (t, J=8.6Hz, 1H), 4.04 (dp, J=11.8, 4.1, 2.9Hz, 2H), 2.42 (s, 3H), 2.28 (d, J=7.6Hz, 2H), 2.00 (s , 3H), 1.96-1.88(m, 2H), 1.86-1.75(m, 3H), 1.73-1.65(m, 1H), 1.57-1.36(m, 7H), 1.31-1.19(m, 2H ), 1.18-1.02 (m, 4H), 0.98 (s, 3H), 0.91 (dq, J=11.5, 6.2Hz, 2H), 0.81 (d, J=6.5Hz, 3H), 0.61 (s, 3H).

[0623] In the third step, 1,2,3,4-tetrahydroisoquinoline-1-one (32.4 mg, 0.22 mmol) was weighed and dissolved in N,N'-dimethylformamide (5.0 mL). The solution was cooled to 0°C in an ice bath, and sodium hydride (22 mg, 0.55 mmol) was added. The solution was then brought to room temperature and stirred for 30 minutes. Finally, 4-methylbenzenesulfonic acid-(3R)-3-[(1R,3aS,3bS,7S,9aR,9] was added. bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]butyl ester (197-2) (60 mg, 0.11 mmol), and then the system was heated to 45 °C and monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, the mixture was cooled to room temperature, quenched with water (10.0 mL), and then extracted with ethyl acetate (6.0 mL * 3). The organic phases were combined, dried, concentrated, and purified by thin-layer chromatography to obtain 2-[(3R)-3-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetrahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]butyl]-1,2,3,4-tetrahydroisoquinoline-1-one (197) (19.7 mg, 0.04 mmol, 36.36%).

[0624] 1 H NMR (399MHz, Chloroform-d) δ8.11-7.99 (m, 1H), 7.35 (dt, J=27.0, 7.5Hz, 2H), 7.14 (d, J=7.3Hz , 1H), 5.33 (d, J=4.9Hz, 1H), 3.66-3.56 (m, 1H), 3.51 (t, J=6.1Hz, 4H), 2.96 (t, J=6.6Hz, 2H), 2. 34-2.13 (m, 2H), 2.04-1.87 (m, 3H), 1.86-1.78 (m, 2H), 1.73 (d, J=3.6Hz, 1H), 1.56 (s, 3H), 1.48 -1.41 (m, 3H), 1.32-1.22 (m, 4H), 1.16 (s, 3H), 1.03 (d, J=6.6Hz, 3H), 0.98 (s, 3H), 0.66 (s, 3H).

[0625] 13CNMR (101MHz, Chloroform-d) δ164.13, 140.76, 137.90, 131.39, 129.72, 128. 16, 127.01, 126.76, 121.64, 77.33, 77.22, 77.02, 76.92, 76.70, 71.77, 56.70, 55.86, 50.05, 45.88, 44.96, 42.38, 42.26, 39.72, 37.22, 36.47, 34.04, 33.56 , 31.86, 31.63, 28.32, 28.23, 24.24, 21.05, 19.39, 18.85, 11.83.LC-MS: [M+H] + =476.30

[0626] Example 199

[0627] Preparation of compound 199(3S,6aS,6bS,9aR,9R,11aS,11bR)-9-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11b-dimethyl-1,2,3,4,5a,6,6a,6b,7,8,9,9a,10,11,11a,11b-hexadecylhydrocyclopenta[1′,2′∶1,2]phenanthro[8a,9-b]oxacyclopropane-3-ol

[0628]

[0629] Cholesterol-6(5)-ene-3β,25-diol (177) (80 mg, 0.179 mmol) was dissolved in dichloromethane (5 mL), and m-chloroperoxybenzoic acid (32.14 mg, 0.19 mmol) was added at room temperature. The mixture was stirred at room temperature for 2 hours and monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was completed, water (10 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (10 mL * 3). The mixture was washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 60:1 ~ 5:1) to obtain (3S, 6aS, 6bS, 9aR, 9R, 11aS, 11bR)-9-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11b-dimethyl-1,2,3,4,5a,6,6a,6b,7,8,9,9a,10,11,11a,11b-hexadecylhydrocyclopenta[1′,2′:1,2]phenanthro[8a,9-b]oxacyclopropane-3-ol (199) (20 mg, 0.043 mmol, 34.78%).

[0630] 1H NMR (399MHz, Chloroform-d) δ3.88 (d, J=13.4Hz, 1H), 2.88 (d, J=4.3Hz, 1H), 2.15-1.99 (m, 2H), 1.91 (s, 3H), 1.73 (d, J=38.2Hz, 2H), 1.51 (d .

[0631] 13 C NMR (100MHz, Chloroform-d) δ109.99, 77.32, 77.20, 77.00, 76.68, 71.11, 68.73, 65 .66, 59.27, 56.82, 56.19, 55.77, 44.36, 42.52, 42.33, 39.84, 39.38, 37.21, 36.37, 3 5.71, 34.83, 32.58, 32.37, 31.07, 29.87, 29.74, 29.30, 29.19, 28.80, 28.08, 24.15, 24.02, 20.78, 20.61, 18.58, 17.03, 15.90, 11.84, 1.00.LCMS: [M-H2O-OH]+=383.30.

[0632] Example 203

[0633] Preparation of compound 203 cholester-5(6)-ene-3β,4a,25-triol

[0634]

[0635] In the first step, (5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (151-7) (780 mg, 1.69 mmol, 1.0 eq.) was weighed and dissolved in dichloromethane (24 mL). Tetrapropylammonium perruthenate (30 mg, 0.09 mmol), N-methylmorpholine oxide (290 mg, 2.46 mmol), and 3A molecular sieve (3.9 g) were added and stirred at room temperature. The mixture was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, the reaction solution was filtered and concentrated. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 20:1 to 10:1) to obtain methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-6-oxoylide-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (203-1) (540 mg, 1.12 mmol, 66.06%), a white solid.

[0636] 1 H NMR (399MHz, CDCl3) δ6.40-6.31 (m, 1H), 5.19 (dd, J=12.6, 7.1Hz, 1H), 3.65 (d, J =0.8Hz, 3H), 2.33-2.18 (m, 3H), 2.15 (d, J = 0.7Hz, 4H), 2.13-2.06 (m, 1H), 2.05-1 .97 (m, 3H), 1.83 (s, 1H), 1.70 (dd, J=19.2, 10.0Hz, 2H), 1.53-1.29 (m, 5H), 1.27 1.18(m, 2H), 1.17-1.00(m, 5H), 0.97(s, 3H), 0.92(d, J=6.5Hz, 3H), 0.66(s, 3H).

[0637] Second step: weighingMethyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-6-oxoylide-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (203-1) (80 mg, 0.17 mmol) was dissolved in tetrahydrofuran (10 mL), and sodium borohydride (13.2 mg, 0.35 mmol, 2.0 eq.) was added at room temperature. The mixture was stirred at room temperature for 16 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, 10 mL of water and 10 mL of ethyl acetate were added for extraction. The mixture was dried, concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate = 20:1~10:1~5:1) to obtain methyl (5R)-5-[(1R,3aS,3bS,6S,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (203-2) (41 mg, 0.085 mmol, 48.48%), a white solid.

[0638] 1 H NMR (399MHz, CDCl3) δ5.87-5.80 (m, 1H), 4.48 (dt, J=10.7, 5.3Hz, 1H), 4.21 (d, J=10.0Hz, 1H), 3.65 (s, 3H), 2.30-2.18 (m, 2H), 2.09 (d, J=1.4Hz, 3H), 1.98 (d, J=12.4Hz, 1H), 1.92- 1.78 (m, 3H), 1.72-1.62 (m, 2H), 1.52 (s, 1H), 1.43 (dt, J=20.6, 5.6Hz, 6H), 1.25 (d, J=11. 9Hz, 2H), 1.17-1.02 (m, 5H), 1.00 (s, 3H), 0.91 (d, J=6.5Hz, 3H), 0.81 (s, 1H), 0.66 (s, 3H).

[0639] In the third step, methyl (5R)-5-[(1R,3aS,3bS,6S,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (203-2) (12 mg, 0.029 mmol) was weighed and dissolved in tetrahydrofuran (5 mL). The solution was cooled with petroleum ether and dry ice, and then a tetrahydrofuran solution of methyl lithium (C: 1.6 M, 0.29 mmol, 10.0 eq.) was added dropwise at low temperature. The solution was slowly heated to room temperature and stirred for 16 hours. The reaction was detected by TLC (dichloromethane:methanol = 20:1). After the reaction was completed, 10 mL of water and 10 mL of ethyl acetate were added for extraction. After drying and concentration, the crude product was purified by column chromatography (dichloromethane:methanol = 300:1 to 100:1) to obtain cholester-5(6)-ene-3β,4α,25-triol (203) (5.3 mg, 0.011 mmol, 36.58%) as a white solid. 1 H NMR (399MHz, CDCl3) δ5.71 (dt, J=5.0, 2.2Hz, 1H), 4.00-3.92 (m, 1H), 3.18 (td, J= 10.4, 9.6, 4.4Hz, 1H), 2.07-1.92 (m, 2H), 1.80 (ddd, J=18.9, 11.0, 4.2Hz, 3H), 1. 62-1.46 (m, 3H), 1.45-1.25 (m, 9H), 1.22 (d, J=12.1Hz, 3H), 1.15 (s, 6H), 1.09-0. 99 (m, 4H), 0.97 (s, 3H), 0.92 (d, J = 6.2Hz, 1H), 0.88 (d, J = 6.5Hz, 3H), 0.63 (s, 3H). 13 C NMR (100MHz, Chloroform-d) δ141.91, 117.78, 71.04, 56.66, 56.01, 49.87, 49.66, 49.44, 49.23, 49.02, 48.8 1, 44.25, 42.20, 39.66, 37.89, 36.39, 35.70, 31.50, 31.35, 28.20, 27.89, 24.21, 20.84, 20.73.LC-MS: [M+Na] + =441.5.

[0640] The two-dimensional spectrum of compound 203 is as follows: Figures 58-59 As shown.

[0641] Example 213

[0642] Preparation of compound 21324-(hydroxycyclopropyl)cholan-6(5)-en-3β-ol

[0643]

[0644]

[0645] Step 1: (1R,3aR,5aR,7S,9aS,11aR)-3a,6,6,9a,11a-pentamethyl-1-[(2R)-6-methylhept-5-en-2-yl]-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (10.00 g, 23.4 mmol, 1.0 eq) was dissolved in dichloromethane (250 mL). After complete dissolution, acetic anhydride (6.7 mL, 71.0 mmol, 3.0 eq), DMAP (0.57 g, 4.7 mmol, 0.2 eq), and TEA (16.3 mL, 117.2 mmol, 5.0 eq) were added sequentially to the reaction system. After the addition was complete, the mixture was stirred at room temperature for 2 hours. After the reaction was monitored to be complete by TLC (petroleum ether:ethyl acetate = 5:1), it was extracted with methanol (10 mL). The reaction solution was washed once with saturated sodium bicarbonate (~100 mL) and once with water (~100 mL), dried over anhydrous sodium sulfate, concentrated, and methanol (~100 mL) was added when the solution was almost dry. The mixture was stirred in an ice bath for 30 minutes, filtered, and the filter cake was washed with a small amount of methanol. The filter cake was dried to give a white solid acetic acid (1R, 3aR, 5aR). 7S,9aS,11aR)-3a,6,6,9a,11a-pentamethyl-1-[(2R)-6-methylhept-5-en-2-yl]-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (I) (9.00 g, 17.3 mmol, purity 90.0%, yield 82.54%). 1 H NMR (400MHz, Chloroform-d): δ4.50 (dd, J=11.5, 4.5Hz, 1H), 2.69 (s, 1H), 2.09-1.87 (m, 8H), 1.7 7-1.24 (m, 26H), 1.15 (d, J=11.4Hz, 3H), 1.00 (s, 3H), 0.93-0.85 (m, 12H), 0.69 (d, J=2.7Hz, 3H). 13C NMR (400MHz, Chloroform-d): δ203.21, 171.00, 134.37, 134.32, 99.99, 80.90, 77.34, 77.02, 76.71, 50.50, 50.30, 49.82, 44.54, 41.14, 37.81, 36.90, 36.08, 36.03, 35.27, 30.95, 30.78, 28.24, 28.15, 27.91, 26.38, 24.23, 24.17, 21.33, 20.98, 19.19, 18.46, 18.40, 18.11, 16.53, 15.78.

[0646] In the second step, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-3a,6,6,9a,11a-pentamethyl-1-[(2R)-6-methylhept-5-en-2-yl]-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (I) (15.00 g, 32.0 mmol, 1.0 ea) was dissolved in dichloromethane (300 mL), purged with nitrogen, and cooled to 0 °C in an ice bath. Ozone was then introduced and the mixture was purged for 10 min. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, the system was purged with nitrogen and concentrated to dryness. The crude product was purified by rapid chromatography (petroleum ether: ethyl acetate = 95:5 to 90:10) to obtain a white solid acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-4-formylbutyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoyl-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (II) (6.50 g, 13.2 mmol, purity 95.5%, yield 41.3%). 1 H NMR (400MHz, Chloroform-d): δ9.77 (t, J=1.9Hz, 1H), 4.50 (dd, J=11.6, 4.5Hz, 1H), 2.51-2.30 (m, 2H), 2.09-1.88 (m, 8H), 1.86-1.24 (m, 17H), 1.23-1.12 (m, 2H), 0.99 (d, J=11.2Hz, 3H), 0.93-0.84 (m, 12H).

[0647] In the third step, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-4-formylbutyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoyl-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (II) (1.50 g, 3.4 mmol, 1.0 eq) was dissolved in dichloromethane (50 mL), and triethylamine (1.41 mL, 10.2 mmol, 3.0 eq) and tert-butyldimethoxytrifluoromethanesulfonate (1.30 g, 5.1 mmol, 1.5 eq) were added sequentially at 0 °C. The reaction solution was stirred at 0°C for 2 hours. TLC (petroleum ether:dichloromethane = 3:1) was used to monitor the reaction until complete. The reaction solution was concentrated to dryness, and 50 mL of n-hexane was added. The mixture was stirred for 10 minutes, filtered, and the filtrate was evaporated to dryness. 50 mL of dichloromethane was added, and the reaction solution was cooled to -78°C. Ozone was bubbled through the solution for 10 minutes. TLC (petroleum ether:dichloromethane = 3:1) was used to monitor the disappearance of the starting material spot. PPh3 (0.89 g, 3.4 mmol, 1.0 eq) was added, and the mixture was stirred for 30 minutes. The reaction solution was directly concentrated and purified by column chromatography. The reaction was carried out in petroleum ether:ethyl acetate = 90:10 to give a white solid acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-1-formylpropyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanohydro-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (III) (300 mg, 0.560 mmol, 16.52%). 1 H NMR (400MHz, Chloroform-d) δ9.76 (dd, J=3.3, 1.1Hz, 1H), 4.50 (dd, J=11.6, 4.5Hz, 1H), 2.47 (dd, J=16.4, 2.5Hz, 1H), 2.17(m, 1H), 2.04(m, 7H), 1.92(m, 1H), 1.43(m, 18H), 0.99(dd, J=9.3, 2.8Hz, 5H), 0.90(m, 12H), 0.74(s, 2H).

[0648] In the fourth step, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-1-formylpropyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (III) (300 mg, 0.700 mmol, 1.0 eq) was dissolved in dichloromethane (10 mL), and ethyl(triphenylphosphine) acetate (243.81 mg, 0.700 mmol, 1.0 eq) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored to be complete by TLC (petroleum ether: dichloromethane = 3:1). The reaction solution was directly concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 95:5) to give a white solid (2E,5R)-5-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetyloxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid ethyl ester (IV) (250 mg, 0.401 mmol, 57.30%). 1 H NMR (400MHz, Chloroform-d) δ6.96 (m, 1H), 5.81 (d, J=15.5Hz, 1H), 4.50 (dd, J=11.5, 4.5Hz, 1H), 4.19 (q, J=7.1Hz, 2H) , 1.99 (m, 8H), 1.63 (m, 12H), 1.31 (m, 6H), 1.17 (m, 3H), 1.00 (s, 3H), 0.92 (t, J=5.3Hz, 4H), 0.87 (m, 10H), 0.69 (m, 3H). 13 C NMR (101MHz, Chloroform-d) δ171.01, 148.45, 134.35, 122.47, 80.90, 77.33, 77.21, 77.01, 76.69, 60.12, 50.50, 50.24, 49.86, 44.60, 39.33, 37.81, 36.91, 36.38, 35.26, 30.80, 28.22, 27.91, 26.92, 26.38, 24.25, 24.17, 21.33, 20.96, 19.19, 18.96, 18.11, 16.53, 15.78, 14.29, -0.01.

[0649] Step 5: (2E,5R)-5-[(1R,3aR,5aR,7S,9aS,11aR)-7-hydroxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-2-enoic acid ethyl ester (IV) (250 mg, 0.55 mmol, 1.0 eq) was dissolved in methanol (10 mL), and palladium (palladium on carbon) (30 mg, 0.282 mmol) was added. The mixture was stirred for 3 hours under a hydrogen atmosphere, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was complete, palladium on carbon was filtered off with diatomaceous earth, the filtrate was concentrated to dryness, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 80:20, phosphomolybdic acid plate) to give a white solid (5R)-5-[(1R,3aR,5aR,7S,9aS,11aR)-7-hydroxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid-1H-cyclopenta[12-a]phenanthrene-1-yl]hexanoic acid ethyl ester (V) (200 mg, 0.392 mmol, 71.68%). 1 H NMR (400MHz, Chloroform-d) δ4.50 (dd, J=11.5, 4.5Hz, 1H), 4.13 (q, J=7.1Hz, 2H), 2.26 (dd, J=15.5, 8.6Hz, 2H), 2.04 (m, 8H), 1.68 (m, 7H), 1.50-1.37 (m, 9H), 1.26 (t, J=7.1Hz, 3H), 1.15 (m, 3H), 1.00 (s, 3H), 0.90 (m, 12H), 0.68 (s, 3H). 13 C NMR (101MHz, Chloroform-d) δ173.95, 171.03, 80.95, 77.33, 77.22, 77.02, 76.70, 60.16, 50.52, 50.26, 49.82, 44.49, 37.82, 36.91, 36.2 1, 35.69, 35.28, 34.82, 30.96, 30.80, 28.16, 27.92, 26.39, 24.25, 24 .18, 21.85, 21.34, 21.00, 19.19, 18.61, 18.13, 16.54, 15.75, 14.28,

[0650] In step six, compound (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (V) (50 mg, 0.103 mmol) was dissolved in THF (50 mL), and tetraisopropyl titanate (48 mg, 0.164 mmol) was added at room temperature. Nitrogen gas was exchanged three times, and ethyl magnesium bromide (1M, 1.03mL, 1.03mmol) was added dropwise at room temperature. TLC (petroleum ether: ethyl acetate = 10:1) was used for monitoring. A small amount of raw material remained. Salt water was added to quench the residue, and ethyl acetate was added for extraction. After separation of the organic phase, the product was dried with anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (petroleum ether: ethyl acetate = 30:1 to 1:1) to obtain 24-(hydroxycyclopropyl)cholan-6(5)-en-3β-ol (213) (3.0mg, 0.006mmol, 6.02%). 1 H NMR (399MHz, Chloroform-d) δ5.33 (d, J=5.1Hz, 1H), 3.51 (dt, J=11.3, 6.4Hz, 1H), 2.43-2.30 (m , 1H), 2.30-2.17(m, 2H), 1.97(t, J=16.1Hz, 2H), 1.85-1.77(m, 3H), 1.58-1.44(m, 8H), 1.39(dd, J=17.3, 7.2Hz, 4H), 1.25 (d, J=10.0Hz, 3H), 1.15-1.03 (m, 4H), 1.03-0.96 (m, 3H), 0.92 (d, J=6. 4Hz, 4H), 0.84-0.76 (m, 1H), 0.71 (q, J=4.5Hz, 2H), 0.66 (d, J=2.8Hz, 3H), 0.42 (q, J=3.8Hz, 2H). 13 C NMR (100MHz, Chloroform-d) δ212.10, 140.74, 121.67, 77.36, 77.32, 77.04, 77.00, 76.71, 76.68, 71.78, 56.70, 55.75, 50.05, 42.88, 42.29, 42.27, 39.70, 37.21, 36.47, 35.85, 35.61, 35.51, 31.86, 31.64, 29.68, 28.18, 24.24, 21.04, 20.43, 19.38, 18.58, 11.81, 7.85LC-MS: [M-OH] + =383.30

[0651] Example 214

[0652] Preparation of compound 214 cholester-6(5)-ene-3β,19,25-triol

[0653]

[0654]

[0655] In the first step, methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (151-6) (1 g, 2.249 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL), and perchloric acid (0.727 mL, 9.00 mmol, 75%, 4.0 eq) and water (1 mL) were added. The reaction system was cooled to below 10 °C, and a tetrahydrofuran solution of N-bromosuccinimide (1.60 g, 9.00 mmol, 4.0 eq) was added. The reaction system was protected from light and stirred in an ice-water bath for 1.5 h. The reaction was then cooled to room temperature, and the completeness of the reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). The reaction was quenched with saturated sodium sulfite solution, and the product was extracted with water (15 mL) and ethyl acetate (10 mL × 3). The product was dried and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 90:10 to 88:12) to give a white solid (5R)-5-[(1R,3aS,3bS,5aR,5R,7S,9aR,9bS,11aR)-7-acetoxy-5a-bromo-5-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopentano[1,2.a]phenanthrene-1-yl]hexanoate (214-1) (400 mg, purity 90%, yield 29.6%). 1 H NMR (400MHz, CDCl3) δ5.47 (dt, J=16.2, 5.5Hz, 1H), 4.21-4.16 (m, 1H), 3.67 (s , 3H), 2.50 (dd, J=13.5, 10.5Hz, 1H), 2.35-2.16 (m, 4H), 2.03 (d, J=3.5Hz, 3H) , 2.01-1.94(m, 2H), 1.88-1.65(m, 6H), 1.60-1.53(m, 5H), 1.46-1.39(m, 2H), 1.30 (d, J=12.2Hz, 4H), 1.22-1.04 (m, 5H), 0.93 (d, J=6.5Hz, 3H), 0.67 (s, 3H).

[0656] In the second step, methyl (5R)-5-[(1R,3aS,3bS,5aR,5R,7S,9aR,9bS,11aR)-7-acetoxy-5a-bromo-5-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (214-1) (400 mg, 0.74 mmol, 1.0 eq) was dissolved in cyclohexane (12 mL) and benzene (3 mL). Diethyl iodophenyl ester (359.08 mg, 1.11 mmol, 1.5 eq) and iodine (224.95 mg, 0.89 mmol, 1.2 eq) were added. The reaction system was sonicated at 45 °C and 40 kHz for 1 h. The reaction progress was monitored by TLC (petroleum ether:ethyl acetate), and the starting materials were consumed. The reaction was quenched with saturated sodium sulfite (100 mL), extracted with ethyl acetate (50 mL × 3), and the combined organic phases were washed with water (30 mL). The combined organic phases were dried and concentrated to obtain the crude product. The crude product was subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 92:8) to give a white solid (5R)-5-[(1R,2S,5R,6R,9S,10S,12R,13R,15S)-15-acetoxy-13-bromo-5-methyl-19-oxapentane[10.5.2.01,13.05,9.02,10]nonadecan-6-yl]hexanoate methyl ester (214-2) (240 mg, purity 95%, yield 57.2%). 1 H NMR (400MHz, CDCl3) δ5.19 (dt, J=15.8, 5.4Hz, 1H), 4.06 (d, J=4.5Hz, 1H), 3.92 (d, J=8.4Hz, 1H), 3.74 (d, J=8.3Hz, 1H), 3.66 (s, 3H), 2.37-2.18 (m, 5H), 2.03 (s, 4H), 2 .01-1.95 (m, 3H), 1.89-1.77 (m, 2H), 1.65 (tdd, J=21.4, 10.5, 4.9Hz, 6H), 1.55-1.4 5(m, 8H), 1.41-1.36(m, 3H), 1.28-1.01(m, 9H), 0.91(d, J=6.6Hz, 3H), 0.69(s, 3H).

[0657] In the third step, the reactant (5R)-5-[(1R,2S,5R,6R,9S,10S,12R,13R,15S)-15-acetoxy-13-bromo-5-methyl-19-oxapentane[10.5.2.01,13.05,9.02,10]nonadecan-6-yl]hexanoate (214-2) (240 mg, 0.445 mmol, 1.0 eq) was dissolved in isopropanol (10 mL), and zinc powder (145.41 mg, 2.224 mmol, 5.0 eq) and acetic acid (0.356 mL, 6.227 mmol, 14 eq) were added. The reaction system was stirred at 90 °C for 3 h. The reaction progress was monitored by TLC plate (petroleum ether:ethyl acetate = 5:1). The reaction was stopped when the reactants were completely consumed. The zinc powder was removed by filtration, the organic phase was collected, and evaporated to dryness. The obtained oily substance was dissolved in ethyl acetate (10 mL), water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phase was collected to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 90:10 to 89:11) to give a white solid (5R)-5-[(1R,3aS,3bS,7S,9aS,9bS,11aR)-7-acetoxy-9a-(hydroxymethyl)-11a-methyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (214-3) (170 mg, purity 95%, yield 76.5%). 1 H NMR (400MHz, CDC13) δ5.80-5.75 (m, 1H), 4.70-4.59 (m, 1H), 3.83 (d, J=11.5Hz, 1H), 3.66 (s, 3H) , 3.62 (d, J=11.4Hz, 1H), 2.42 (ddd, J=12.9, 4.8, 2.1Hz, 1H), 2.33-2.21 (m, 3H), 2.03 (s, 3H), 1. 95(dt, J=13.8, 3.5Hz, 1H), 1.87-1.76(m, 3H), 1.73-1.43(m, 9H), 1.42-1.32(m, 3H), 1.26(dd, J =14.5, 7.1Hz, 3H), 1.18-1.02 (m, 5H), 0.93 (d, J = 6.6Hz, 3H), 0.87 (d, J = 6.1Hz, 1H), 0.73 (s, 3H).

[0658] In the fourth step, the reactant (5R)-5-[(1R,3aS,3bS,7S,9aS,9bS,11aR)-7-acetoxy-9a-(hydroxymethyl)-11a-methyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (214-3) (30 mg, 0.065 mmol)] was dissolved in tetrahydrofuran (10 mL). After cooling the reaction system to 0 °C, methyl magnesium chloride (0.130 mL, 0.391 mmol, 3 mol / L) was slowly added, and the reaction system was stirred at room temperature for 2 h. The reaction progress was monitored by TLC plate (petroleum ether:ethyl acetate = 5:1). The reaction was stopped when the reactants were completely consumed. 20 mL of water was added to the reaction system, and the mixture was extracted with ethyl acetate (25 mL × 3). The organic phase was collected, dried with anhydrous sodium sulfate, and then evaporated under vacuum to obtain the crude product. The crude product was dissolved in EA and subjected to column chromatography (petroleum ether: ethyl acetate = 55:45 to 53:47) to give cholesterol-6(5)-ene-3β,19,25-triol (214) (8.70 mg, purity 97%, yield 31.2%). 1 H NMR (400MHz, CDCl3) δ5.75 (d, J=5.0Hz, 1H), 3.82 (d, J=11.4Hz, 1H), 3.64-3.53 (m, 2H), 2.38 (ddd, J= 12.9, 4.6, 2.3Hz, 1H), 2.19 (t, J=11.8Hz, 1H), 2.07-1.99 (m, 2H), 1.93 (dd, J=13.8, 3.4Hz, 1H), 1.83 (dt, J=10.3, 7.8Hz, 3H), 1.63 (dd, J=12.9, 3.7Hz, 1H), 1.59-1.51 (m, 4H), 1.45 (d, J=4.1Hz, 4H), 1.3 8 (d, J=13.4Hz, 3H), 1.25 (s, 3H), 1.21 (s, 6H), 1.13-1.03 (m, 4H), 0.93 (d, J=6.5Hz, 3H), 0.73 (s, 3H). 13 C NMR (101MHz, CDCl3) δ135.51, 127.45, 71.40, 71.13, 62.75, 57.66, 56.04, 50.38, 44.42, 42.56, 42.33, 41.53, 40.04, 3 6.44, 35.73, 33.43, 31.99, 31.24, 29.70, 29.38, 29.19, 28.26, 24.08, 21.80, 20.76, 18.67, 12.23.LC-MS: [M-OH-H2O] + =383.30.

[0659] Example 216

[0660] Preparation of compound 216 3β,25-dihydroxy-5α-cholesterol-7-one

[0661]

[0662] In the first step, cholesterol-6(5)-ene-3β,25-diol (177) (300 mg, 0.745 mmol) was dissolved in dichloromethane (30 mL) under nitrogen protection. Triethylamine (0.52 mL, 3.73 mmol), 4-dimethylaminopyridine (18.2 mg, 0.15 mmol), and acetic anhydride (0.084 mL, 0.89 mmol) were added and reacted at room temperature for 2 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1:1). After the reaction was completed, saturated ammonium chloride was added to quench the reaction solution, followed by extraction with dichloromethane and separation of the organic phase. The solution was dried with anhydrous sodium sulfate, and the crude product obtained by concentrating the reaction solution was purified by column chromatography (petroleum ether: ethyl acetate = 60:1 to 5:1) to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (216-1) (200 mg, 0.41 mmol, 54.33%), a white solid. 1 H NMR (399MHz, Chloroform-d) δ5.35 (d, J=5.0Hz, 1H), 4.57 (q, J=6.4, 5.3Hz, 1H), 2.29 (d, J=7.9Hz, 2H), 2.01 (s, 3H), 1.95 (d, J=18.2Hz, 2H), 1.84 (d, J=11.6Hz, 2H), 1.61-1.51 (m, 4H), 1.491.30 (m, 8H), 1.19 (s, 6H), 1.13-1.04 (m, 3H), 0.91 (d, J=6.5Hz, 3H), 0.66. (s, 3H).

[0663] In the second step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (216-1) (280 mg, 0.63 mmol) was dissolved in acetone (10 mL), and cobalt acetate (1.11 mg, 0.006 mmol), N-hydroxyphthalimide (10.27 mg, 0.063 mmol) and tert-butyl hydroperoxide (226.97 mg, 2.52 mmol) were added at room temperature. The mixture was stirred at room temperature for 40 hours and monitored by TLC (petroleum ether:ethyl acetate = 3:1). After the reaction was completed, water (10 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (10 mL * 3). The mixture was washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 60:1 to 4:1) to give acetate-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-oxonyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (216-2). (180 mg, 0.37 mmol, 59.21%). 1 H NMR (399MHz, Chloroform-d) δ5.68 (s, 1H), 4.69 (s, 1H), 2.57-2.34 (m, 3H), 2.21 (s, 1H), 2.03 (s, 3H), 1.94 (d, J=13.6Hz, 3H), 1.86 (t, J=8.9Hz, 1H), 1.73-1.62 (m, 1H), 1.57 (s, 4H ), 1.51 (dd, J=13.4, 6.2Hz, 2H), 1.44-1.39 (m, 3H), 1.35 (dd, J=11.9, 6.3Hz, 3H), 1.28-1. 25 (m, 2H), 1.19 (s, 9H), 1.06 (dd, J=15.2, 9.2Hz, 2H), 0.92 (d, J=6.5Hz, 3H), 0.66 (s, 3H).

[0664] In the third step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-oxonyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (216-2) (140 mg, 0.31 mmol) was dissolved in ethyl acetate (10 mL), palladium / carbon (30 mg, 0.28 mmol) was added at room temperature, hydrogen was purged, and the mixture was stirred at room temperature for 1 hour. The mixture was monitored by TLC (petroleum ether:ethyl acetate = 2:1). After the reaction was completed, the product was filtered and evaporated to dryness to obtain 150 mg of crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 60:1 to 5:1) to give acetic acid-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-oxoylidenehexadecyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (216-3) (100 mg, 0.21 mmol, 67.56%). 1 H NMR (399MHz, Chloroform-d) δ4.67 (s, 1H), 2.29 (d, J=15.5Hz, 2H), 2.16 (s, 1H), 2.00 (s, 3H), 1. 94 (s, 1H), 1.88 (d, J = 19.1Hz, 2H), 1.77 (d, J = 13.7Hz, 1H), 1.64 (d, J = 16.8Hz, 1H), 1.58 (s, 4H), 1 .56-1.44(m, 5H), 1.44-1.39(m, 2H), 1.36(d, J=10.8Hz, 4H), 1.25(d, J=11.1Hz, 2H), 1.19(s, 6H) , 1.11 (d, J=7.2Hz, 1H), 1.08 (s, 3H), 1.03 (d, J=12.5Hz, 2H), 0.90 (d, J=6.5Hz, 3H), 0.63 (s, 3H).

[0665] In the fourth step, acetic acid-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-oxoylhexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (216-3) (30 mg, 0.065 mmol) was dissolved in a mixed solvent of tetrahydrofuran (3 mL) and methanol (1.5 mL), and lithium hydroxide (0.5 mL, 0.5 mmol) was added at room temperature. The mixture was stirred at room temperature for 2 hours and monitored by TLC (petroleum ether:ethyl acetate = 2:1). After the reaction was completed, the crude product was concentrated to obtain 50 mg of crude product, which was purified by column chromatography (dichloromethane:methanol = 80:1) to obtain 3β,25-dihydroxy-5α-cholest-7-one (216) (13.8 mg, 0.031 mmol, 48.09%) as a white solid. 1 H NMR (399MHz, Chloroform-d) δ3.63-3.54 (m, 1H), 2.40-2.26 (m, 2H), 2.18 (s, 1H), 2.03-1.94 (m, 2H), 1.85 (d, J=5.9Hz, 2H), 1.75 (d, J=13.4Hz, 1H), 1.52(d, J=11.7Hz, 7H), 1.48-1.39(m, 6H), 1.37(d, J=10.2Hz, 5H), 1.19(s , 6H), 1.06 (s, 3H), 1.01-0.93 (m, 2H), 0.90 (d, J=6.5Hz, 3H), 0.63 (s, 3H). 13 C NMR (100MHz, cdcl3) δ212.08, 77.30, 77.17, 76.98, 76.66, 71.08, 70.63, 55.19, 54.91, 49.97, 48.85, 46.80, 46.06, 44 .35, 42.48, 38.71, 37.85, 36.36, 35.95, 35.59, 31.02, 29.29, 29.20, 28.40, 21.82, 20.70, 18.75, 11.99.LC-MS: [M-OH] + =401.35.

[0666] Example 217

[0667] Preparation of compound 217 24-[cyclopropyl(hydroxy)methyl]-5α-cholan-3β-ol

[0668]

[0669] In the first step, the reactant (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate methyl ester (151-6) (2 g, 4.50 mmol, 1.0 eq) was dissolved in methanol (50 mL), and concentrated sulfuric acid (98%) (1 mL, 18.76 mmol) was added dropwise. The mixture was then heated to reflux and stirred for 1 hour. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was complete, most of the methanol was concentrated off the reaction solution, ethyl acetate was added, and the mixture was washed twice with saturated sodium bicarbonate and once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and the crude product was concentrated to dryness. The crude product was then purified by rapid silica gel column chromatography (petroleum ether:ethyl acetate = 90:10 to 80:20, phosphomolybdic acid plate) to give a white solid (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate methyl ester (217-1) (1.8 g, 4.024 mmol, 89.46%). 1 H NMR (400MHz, CDCl3) δ5.37-5.33 (m, 1H), 3.67 (s, 3H), 3.52 (dd, J=7.7, 3.3Hz, 1H), 2.35-2.20 (m, 4H), 2.03-1.93 (m, 2H), 1.84 (ddd, J= 12.5, 7.5, 4.3Hz, 3H), 1.75-1.60 (m, 4H), 1.59-1.35 (m, 9H), 1.33-1.04 (m, 7H), 1.03-0.99 (m, 3H), 0.93 (d, J=6.6Hz, 3H), 0.68 (s, 3H).

[0670] In the second step, the reactant (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (217-1) (1.7 g, 4.38 mmol) was dissolved in DMF (50 mL), and TBSCl (3.3 g, 21.9 mmol, 5.0 eq) and imidazole (0.30 g, 4.38 mmol, 1.0 eq) were added sequentially. The reaction was heated to 100 °C, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was completed, the reaction solution was cooled to room temperature, ethyl acetate (100 mL) was added, and the mixture was washed three times with water. The organic phase was then dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by rapid column chromatography (petroleum ether: ethyl acetate = 90:10) to give a colorless waxy compound (5R)-5-[(1R,3aR,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-3b,9a,11a-trimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate methyl ester (217-2) (1.9 g, 3.40 mmol, 77.73%). 1 H NMR (400MHz, CDCl3) δ5.26-5.18 (m, 1H), 3.57 (s, 3H), 3.43-3.33 (m, 1H), 2.16 (d, J=1.9Hz, 4H), 1.93-1.82 (m, 2H), 1.65-1.55 (m, 2H), 1.39 (d, J=3.0Hz, 9H), 0.99 (s, 7H), 0.90 (s, 5H), 0.85-0.79 (m, 21H), 0.57 (s, 3H), -0.00 (s, 6H).

[0671] In the third step, the reactant (5R)-5-[(1R,3aR,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-3b,9a,11a-trimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (217-2) (1.8 g, 3.58 mmol, 1.0 eq) was dissolved in tetrahydrofuran (50 mL), and lithium aluminum hydride (0.168 g, 4.269 mmol, 1.2 eq) was added. After stirring at room temperature for 30 min, the reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was completed, the reaction solution was quenched with sodium sulfate decahydrate, filtered, and the filtrate was concentrated to obtain the crude product. Subsequently, the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 80:20 to 70:30) to give a white solid (5R)-5-[(1R,3aR,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-3b,9a,11a-trimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-1-ol (217-3) (1.7 g, 3.22 mmol, 90.01%). 1 H NMR (400MHz, CDCl3) δ5.26 (d, J=5.3Hz, 1H), 3.59 (t, J=6.6Hz, 2H), 3.473.37 (m, 1H), 2.27-2 .17 (m, 1H), 2.11 (ddd, J=13.4, 5.0, 2.2Hz, 1H), 1.93 (s, 2H), 1.77 (s, 2H), 1.66 (d, J=12.2Hz, 1H), 1.47(ddd, J=13.9, 8.7, 4.5Hz, 7H), 1.37-1.29(m, 3H), 1.25-1.15(m, 2H), 1.13-0.97(m , 5H), 0.94 (s, 3H), 0.87 (d, J=6.3Hz, 4H), 0.840.81 (m, 10H), 0.62 (s, 3H), 0.02-0.02 (m, 6H).

[0672] In the fourth step, compound (5R)-5-[(1R,3aR,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-3b,9a,11a-trimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-1-ol (217-3) (1.0 g, 2.11 mmol, 1.0 eq) was dissolved in DCM (100 mL), and Dysmartin oxidant (1.068 g, 2.532 mmol, 1.2 eq) was added. The mixture was stirred at room temperature, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, the reaction solution was quenched with saturated sodium sulfite, washed with saturated sodium bicarbonate, dried with anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 90:10 to 80:20) to give a white solid (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanal (217-4) (0.6 g, 1.142 mmol, 54.23%). 1 H NMR (400MHz, CDCl3) δ9.71 (t, J=1.8Hz, 1H), 5.26 (d, J=5.3Hz, 1H), 3.49-3. 36(m, 1H), 2.34(d, J=6.6Hz, 2H), 2.25-2.07(m, 2H), 1.93(s, 2H), 1.81-1.60 (m, 4H), 1.481.30 (m, 8H), 1.03 (ddd, J=14.6, 12.0, 6.3Hz, 7H), 0.94 (s, 4H) , 0.89 (d, J=6.5Hz, 4H), 0.85-0.81 (m, 9H), 0.62 (s, 3H), 0.02--0.02 (m, 7H).

[0673] In the fifth step, (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanal (217-4) (100 mg, 0.205 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL), cooled to 0 °C in an ice bath, and after complete dissolution, cyclopropylmagnesium bromide (2.054 mL, 1 mol / L, 2.054 mmol, 10 eq) was added to the reaction system. After the addition was complete, the mixture was stirred at room temperature for 2 hours. After the reaction was monitored to be complete by TLC (petroleum ether:ethyl acetate = 5:1), the reaction mixture was extracted with saturated ammonium chloride solution (10 mL) under ice bath conditions. The reaction solution was washed with water (10 mL x 3) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 100:0). Purification of t094∶6) yielded (5R)-1-cyclopropyl-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-1-ol (217-5) (72 mg, 0.109 mmol, purity 80.0%, yield 53.02%). 1 H NMR (400MHz, CDCl3) δ5.31 (d, J=5.3Hz, 1H), 3.47 (td, J=10.9, 5.4Hz, 1H), 2.88-2.82 (m, 1H), 2.27 (t, J=11.1Hz, 1H), 2.18-2.13 (m, 1H) 1.99 (s, 2H), 1.82 (s, 2H), 1.71 (d, J=12.3Hz, 1H), 1 .61-1.51 (m, 11H), 141 (dd, J=12.4, 6.8Hz, 3H), 1.27 (d, J=12.0Hz, 3H), 1.16-1.01 (m, 5H), 0. 96-0.84 (m, 15H), 0.67 (s, 3H), 0.58-0.44 (m, 2H), 0.24 (d, J=4.7Hz, 2H), 0.02--0.02 (m, 5H).

[0674] The sixth compound, (5R)-1-cyclopropyl-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-1-ol (217-5) (72 mg, 0.136 mmol, 1.0 eq), was dissolved in methanol (5 mL) and tetrahydrofuran (2 mL), and palladium on carbon was added. The mixture was then purged three times with hydrogen. The reaction was monitored by ¹H NMR. After the reaction was complete, palladium carbon was filtered through diatomaceous earth, the organic phases were combined, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 90:10 to 75:25) to give a white solid 24-[cyclopropyl(hydroxy)methyl]-5α-cholan-3β-ol (217) (17.88 mg, 0.122 mmol, purity 94.86%, yield 89%). 1 H NMR (400MHz, CDCl3) δ3.63-3.54 (m, 1H), 2.85 (td, J=7.8, 4.9Hz, 1H), 1.96 (dt, J=12.6, 3.4Hz, 1H), 1.86-1.75 (m, 2H), 1.74-1.58 (m, 3H), 1.49 -1.19 (m, 14H), 1.03 (ddd, J=22.5, 17.2, 8.7Hz, 7H), 0.90 (t, J=7.6Hz, 5H), 0.80 (s, 3H), 0.65 (s, 4H), 0.57-0.44 (m, 2H), 0.24 (d, J=4.7Hz, 2H). 13 C NMR (101MHz, CDCl3) δ71.40, 64.92, 63.38, 56.51, 56.09, 54.40, 45.03, 42.62, 39.97, 38.29, 37.56, 36.91, 35.76, 3 5.39, 32.09, 31.44, 28.75, 28.10, 24.22, 21.26, 19.64, 18.67, 18.01, 12.28, 11.99, 3.35, 2.27.LC-MS: [M-OH-H2O] + =381

[0675] Example 219

[0676] Preparation of compound 21924-(2,2,2-trifluoro-1-hydroxyethyl)cholan-6(5)-en-3β-ol

[0677]

[0678] (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanal (217-4) (100 mg, 0.205 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL). After complete dissolution, cesium fluoride (15.6 mg, 0.103 mmol, 0.5 eq) and (trifluoromethyl)trimethylsilane (146.03 mg, 0.513 mmol, 5.0 eq) were added sequentially to the reaction system. After the addition was complete, the mixture was stirred at room temperature for 2 hours. After a large polarity point was detected by TLC (petroleum ether: ethyl acetate = 10:1), tetrabutylammonium fluoride (2 ml, 1 mol / L, 10.0 eq) was added and stirred at room temperature for 1 hour. A large polarity point was detected by TLC (petroleum ether: ethyl acetate = 2:1). The reaction mixture was quenched with saturated ammonium chloride solution (50 mL) under ice bath conditions. The reaction mixture was washed with water (10 ml x 3) and extracted with ethyl acetate (10 ml x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 90:10 to 75:25) to give 24-(2,2,2-trifluoro-1-hydroxyethyl)cholan-6(5)-en-3β-ol (219) (10.82 mg, 0.075 mmol, purity 91.82%, yield 36.37%). 1 HNMR (400MHz, CDCl3) δ5.37-5.33 (m, 1H), 3.96-3.87 (m, 1H), 3.52 (ddd, J=15.7, 11.0, 4.5Hz, 1H), 2.27 (tdd, J=12.8, 8.9, 6.6Hz, 2H), 2.00 (d, J=3.2Hz, 3H), 1.84 (dd, J=14.1, 3.4Hz, 3H), 1.61-1.42 (m, 15H), 1.35-1.19 (m, 5H), 1.10 (dd, J=18.5, 5.5Hz, 4H), 0.94 (d, J=6.6Hz, 3H). 13 C NMR (101MHz, CDCl3) δ155.09, 121.69, 115.18, 71.82, 69.47, 56.76, 55.91, 50.11, 42.35, 37.25, 35.58, 31.90, 28.24, 19.40, 11.86 19 F(376MHz, CDCl3)δ-80.04LC-MS: [M-OH] + =425

[0679] Example 220

[0680] Preparation of compound 220 24-[cyclopropyl(hydroxy)methyl]-5α-cholan-3β-ol

[0681]

[0682]

[0683] In the first step, (22E, 24S)-stigmaster-6(5),22(23)-dien-3β-ol (50.0 g, 0.12 mol, 1.0 eq.) and 4-dimethylaminopyridine (2.96 g, 24.23 mmol) were weighed and dissolved in dichloromethane (500 mL). Triethylamine (61.90 g, 605.77 mmol) was added, followed by acetic anhydride (37.11 g, 0.36 mol) under stirring at room temperature. The mixture was then stirred at room temperature for 2 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was completed, the reaction solution was concentrated. The crude product was then added to 300 mL of methanol and stirred. After filtration, the filter cake was washed with 20 mL of methanol twice and then dried to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E)-5-ethyl-6-methylhept-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-1) (53.8 g, 112.4 mmol, 92.77%), a white solid. 1 H NMR (399MHz, Chloroform-d) δ5.35 (d, J=4.9Hz, 1H), 5.14 (dd, J=15.1, 8.5Hz, 1H), 4.99 ( dd, J=15.2, 8.5Hz, 1H), 4.67-4.48 (m, 1H), 2.30 (d, J=7.8Hz, 2H), 2.01 (s, 3H), 2.00-1.9 1 (m, 2H), 1.84 (d, J=11.3Hz, 2H), 1.68 (dd, J=9.4, 4.8Hz, 1H), 1.48 (dt, J=24.5, 9.2Hz, 7 H), 1.28-1.07 (m, 6H), 1.00 (d, J=3.2Hz, 6H), 0.80 (dd, J=20.1, 6.4Hz, 9H), 0.68 (s, 3H).

[0684] The second step involves weighing acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E)-5-ethyl-6-methylhept-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-1) 5.0 g (11 mmol) was dissolved in tetrahydrofuran (200 mL) and water (20.0 mL). Pyridine (2.12 mL, 27.5 mmol), N-methylmorpholine oxide (5.15 g, 43.98 mmol), and potassium osmium tetroxide (7.3 mL, 1.1 mmol) were added at room temperature. The mixture was stirred overnight at room temperature. TLC (petroleum ether:ethyl acetate = 3:1) showed the presence of an intermediate (vicinal diol). Sodium periodate (9.41 g, 43.98 mmol) was then added to the reaction solution at 0 °C, and the mixture was stirred at room temperature for 1 hour. The intermediate was converted to the product by TLC (petroleum ether: ethyl acetate = 3:1). Subsequently, 50 mL of water was added, and the mixture was extracted with 50 mL of ethyl acetate three times. After drying and concentration, the crude product was purified by column chromatography (petroleum ether: ethyl acetate = 60:1) to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aS)-1-[(1S)-1-formylethyl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-2) (0.64 g, 1.6 mmol, 14.73%) as a white solid. 1 HNMR (399MHz, Chloroform-d) δ9.55 (s, 1H), 5.36 (s, 1H), 4.60 (s, 1H), 2.31 (s, 3H), 2.02 (s, 3H), 1.93 (s, 2H), 1.84 (d, J=11.2Hz, 3H), 1.63 (s, 3 H), 1.49 (d, J=10.9Hz, 4H), 1.42-1.19 (m, 3H), 1.15 (s, 1H), 1.11 (d, J=6 .8Hz, 3H), 1.06 (s, 1H), 1.01 (s, 3H), 0.98 (d, J=5.8Hz, 1H), 0.71 (s, 3H).

[0685] The third step involves weighing (methoxymethyl)triphenylphosphine chloride (8.7 g, 25.0 mmol) and dissolving it in tetrahydrofuran (38 mL). The solution is purged with nitrogen three times, cooled to 0 °C, and bis(trimethylsilyl)aminosodium (12.8 mL, 25.0 mmol) is added dropwise. After the addition is complete, the mixture is stirred at 0 °C for half an hour. Then, acetic acid-(1R, 3aS, 3bS, 7S, 9aR, 9bS, l1aS)-1-[(1S)- A solution of 1-formylethyl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-2) (1.9 g, 5.1 mmol) in tetrahydrofuran (18 mL) was prepared, and then stirred at room temperature for 1 hour. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1), and the reaction was complete. Subsequently, 30 mL of water and 15 mL of ethyl acetate were added for extraction, dried, and concentrated. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 100:1 to 30:1) to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2S,3E)-4-methoxybut-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-3) (0.75 g, 1.8 mmol, 34.87%) as a white solid. 1 H NMR (399MHz, Chloroform-d) δ6.22 (d, J=12.6Hz, 1H), 5.71 (d, J=6.4Hz, 0H), 5.35 (d, J=4.1Hz, 1H), 4.57 (m, 2H), 4.15 (m, 0H), 3.52 (d, J = 10.6Hz, 1H), 3.46 (d, J = 8.7Hz, 3H), 2.59 (d, J = 6.6Hz , 0H), 2.30 (d, J=7.6Hz, 3H), 2.01 (s, 3H), 1.95 (d, J=5.8Hz, 3H), 1.84 (d, J=10.9Hz, 3H), 1.69 ( d, J=4.6Hz, 1H), 1.52 (s, 2H), 1.48 (s, 2H), 1.14 (m, 3H), 1.03 (s, 3H), 1.00 (s, 3H), 0.67 (s, 3H).

[0686] In the fourth step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2S,3E)-4-methoxybut-3-en-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-3) (0.75 g, 1.87 mmol) was dissolved in tetrahydrofuran (7.5 mL), and 5N hydrochloric acid (2.25 mL, 11.2 mmol) was added. The mixture was then heated to 50 °C and stirred for 1 hour. The mixture was monitored by TLC (petroleum ether:ethyl acetate = 10:1). Subsequently, 15 mL of water and 10 mL of ethyl acetate were added for extraction, dried, and concentrated. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 50:1 to 20:1) to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-formylpropyl-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-4) (0.52 g, 1.3 mmol, 68.26%) as a white solid. 1 H NMR (399MHz, Chloroform-d) δ9.74 (s, 1H), 5.36 (d, J = 4.4Hz, 1H), 4.60 (s, 1H), 2.44 (d, J = 16 .5Hz, 1H), 2.30 (d, J=6.7Hz, 2H), 2.20-2.10 (m, 1H), 2.01 (s, 3H), 1.95 (d, J=14.4Hz, 2H), 1.8 4(d, J=11.5Hz, 3H), 1.57 (dd, J=19.7, 8.6Hz, 4H), 1.46 (dd, J=14.2, 10.3Hz, 3H), 1.29-1.16 (m, 3H), 1.15-1.03 (m, 3H), 1.01 (d, J = 4.5Hz, 6H), 0.96-0.89 (m, 1H), 0.70 (d, J = 12.6Hz, 3H).

[0687] In the fifth step, 0.52 g (1.34 mmol) of acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-formylpropyl-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-4) was dissolved in tetrahydrofuran (30 mL), and 2.25 g (6.73 mmol) of (triphenyl-λ5-methylphosphine)acetic acid methyl ester was added. The reaction system was heated to 90 °C and stirred for 3 hours. The reaction was monitored by HPLC until it was complete. The sample was then cooled to room temperature, and 15 mL of water and 10 mL of ethyl acetate were added for extraction. After drying and concentration, the crude product was purified by column chromatography (petroleum ether:ethyl acetate = 50:1 to 20:1) to obtain (2E,5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (151-5) (0.44 g, 0.94 mmol, 70.20%) as a white solid. 1 H NMR (399MHz, Chloroform-d) δ6.99-6.86 (m, 1H), 5.80 (d, J=15.6Hz, 1H), 5.36 (s , 1H), 4.59 (s, 1H), 3.71 (s, 3H), 2.30 (d, J=6.7Hz, 3H), 2.01 (s, 3H), 2.00-1.90 ( m, 3H), 1.84 (d, J = 10.9Hz, 3H), 1.54 (d, J = 16.6Hz, 5H), 1.50-1.37 (m, 3H), 1.33- 1.17 (m, 2H), 1.16-1.02 (m, 4H), 1.00 (s, 3H), 0.93 (d, J=6.6Hz, 3H), 0.67 (s, 3H).

[0688] Step 6: Weigh (2E, 5R)-5-[(1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b, 4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (151-5) (0.44 g, 1.0 mmol) and dissolve it in a mixed solvent of tetrahydrofuran (9.0 mL) and methanol (4.5 mL). Add nickel chloride (130 mg, 1.0 mmol), and then add sodium borohydride (56.92 mg, 1.5 mmol) under stirring at room temperature. The reaction solution contains a large amount of... After bubble generation and stirring at room temperature for 1 hour, a sample was taken, concentrated, and monitored by NMR. The reaction of the raw materials was complete. 15 mL of water and 10 mL of ethyl acetate were added for extraction. After drying and concentration, the crude product was purified by column chromatography (petroleum ether:ethyl acetate = 60:1 to 20:1) to obtain methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (151-6) (0.36 g, 0.78 mmol, 77.74%), a white solid. 1 H NMR (399MHz, Chloroform-d) δ5.36 (s, 1H), 4.59 (s, 1H), 3.65 (s, 3H), 2.35-2.18 (m, 4H), 2.01 ( s, 3H), 1.94 (d, J = 15.3Hz, 2H), 1.84 (d, J = 10.9Hz, 3H), 1.68 (s, 1H), 1.55 (s, 2H), 1.53-1.44 (m, 4H), 1.39 (d, J=10.2Hz, 3H), 1.22 (dd, J=22.8, 12.0Hz, 2H), 1.15-1.09 (m, 2H), 1.08-1.03 (m, 2 H), 1.00 (s, 3H), 0.96 (s, 1H), 0.91 (d, J = 6.3Hz, 3H), 0.89-0.79 (m, 1H), 0.64 (d, J = 12.6Hz, 3H).

[0689] In step seven, the reactant (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (151-6) (2 g, 4.50 mmol, 1.0 eq) was dissolved in methanol (50 mL), and concentrated sulfuric acid (98%) (1 mL, 18.76 mmol) was added dropwise. The mixture was then heated to reflux and stirred for 1 hour. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1 plate). After the reaction was complete, The reaction solution was concentrated to remove most of the methanol, ethyl acetate was added, and the mixture was washed twice with saturated sodium bicarbonate and once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was then purified by rapid silica gel column chromatography (PE:EtOAc = 90:10 to 80:20, phosphomolybdic acid plate) to give a white solid (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate methyl ester (220-1) (1.8 g, 4.024 mmol, 89.46%). 1 H NMR (400MHz, Chloroform-d) δ5.37-5.33 (m, 1H), 3.67 (s, 3H), 3.52 (dd, J=7.7, 3.3Hz, 1H), 2.35-2.20 (m, 4H), 2.03-1.93 (m, 2H), 1.84 (ddd .

[0690] In step 8, the reactant (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (220-1) (1.7 g, 4.38 mmol) was dissolved in DMF (50 mL), and TBSCI (3.3 g, 21.9 mmol, 5.0 eq) and imidazole (0.30 g, 4.38 mmol, 1.0 eq) were added sequentially. The reaction was heated to 100 °C, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was completed, the reaction solution was cooled to room temperature, ethyl acetate (100 mL) was added, and the mixture was washed three times with water. The organic phase was then dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by rapid column chromatography (petroleum ether: ethyl acetate = 90:10) to give a colorless waxy compound (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (220-2) (1.9 g, 3.40 mmol, 77.73%). 1 H NMR (400MHz, Chloroform-d) δ5.26-5.18 (m, 1H), 3.57 (s, 3H), 3.43-3.33 (m, 1H), 2.16 (d, J=19Hz, 4H), 1.93-1.82 (m, 2 H), 1.65-1.55 (m, 2H), 1.39 (d, J=3.0Hz, 9H), 0.99 (s, 7H), 0.90 (s, 5H), 0.850.79 (m, 21H), 0.57 (s, 3H), -0.00 (s, 6H).

[0691] In step nine, the reactant (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (220-2) (1.8 g, 3.58 mmol, 1.0 eq) was dissolved in tetrahydrofuran (50 mL), and lithium aluminum hydride (0.168 g, 4.269 mmol, 1.2 eq) was added. After stirring at room temperature for 30 min, the reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was completed, the reaction solution was quenched with sodium sulfate decahydrate, filtered, and the filtrate was concentrated to obtain the crude product. Subsequently, the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 80:20 to 70:30) to give a white solid (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-1-ol (220-3) (1.7 g, 3.22 mmol, 90.01%). 1 H NMR (400MHz, Chloroform-d) δ5.26 (d, J=5.3Hz, 1H), 3.59 (t, J=6.6Hz, 2H), 3.47-3.37 (m, 1H), 2 .27-2.17 (m, 1H), 2.11 (ddd, J=13.4, 5.0, 2.2Hz, 1H), 1.93 (s, 2H), 1.77 (s, 2H), 1.66 (d, J=12.2 Hz, 1H), 1.47 (ddd, J=13.9, 8.7, 4.5Hz, 7H), 1.37-1.29 (m, 3H), 1.25-1.15 (m, 2H), 1.13-0.97 (m , 5H), 0.94 (s, 3H), 0.87 (d, J=6.3Hz, 4H), 0.84-0.81 (m, 10H), 0.62 (s, 3H), 0.02--0.02 (m, 6H).

[0692] Step 10 Compound (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-1-ol (220-3) (1.0 g, 2. 11 mmol (1.0 eq) was dissolved in dichloromethane (100 mL), and Dysmartin oxidant (1.068 g, 2.532 mmol, 1.2 eq) was added. The mixture was stirred at room temperature, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, the reaction solution was quenched with saturated sodium sulfite, washed with saturated sodium bicarbonate, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 90:10). (to 80∶20) yielded a white solid (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanal (220-4) (0.6 g, 1.142 mmol, 54.23%). 1 H NMR (400MHz, Chloroform-d) δ9.71 (t, J=1.8Hz, 1H), 5.26 (d, J=5.3Hz, 1H), 3. 49-3.36 (m, 1H), 2.34 (d, J=6.6Hz, 2H), 2.25-2.07 (m, 2H), 1.93 (s, 2H), 1.81- 1.60 (m, 4H), 1.481.30 (m, 8H), 1.03 (ddd, J=14.6, 12.0, 6.3Hz, 7H), 0.94 (s, 4 H), 0.89 (d, J=6.5Hz, 4H), 0.85-0.81 (m, 9H), 0.62 (s, 3H), 0.02--0.02 (m, 7H).

[0693] In step 11, (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanal (220-4) (100 mg, 0.205 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL), cooled to 0 °C in an ice bath, and after complete dissolution, cyclopropylmagnesium bromide (2.054 mL, 1 mol / L, 2.054 mmol, 10 eq) was added to the reaction system. After the addition was complete, the mixture was stirred at room temperature for 2 hours. After the reaction was monitored to completion by TLC (petroleum ether:ethyl acetate = 5:1), the reaction mixture was extracted with saturated ammonium chloride solution (10 mL) under ice bath conditions. The reaction solution was washed with water (10 mL x 3), extracted with ethyl acetate (10 mL x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (100:0 to 100:1). Purification at 94:6 yielded (5R)-1-cyclopropyl-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-1-ol (220-5) (72 mg, 0.109 mmol, purity 80.0%, yield 53.02%). 1 H NMR (400MHz, Chloroform-d) δ5.31 (d, J=5.3Hz, 1H), 3.47 (td, J=10.9, 5.4Hz, 1H), 2.88-2.82 (m , 1H), 2.27 (t, J=11.1Hz, 1H), 2.18-2.13 (m, 1H), 1.99 (s, 2H), 1.82 (s, 2H), 1.71 (d, J=12.3Hz, 1 H), 1.61-1.51 (m, 11H), 1.41 (dd, J=12.4, 6.8Hz, 3H), 1.27 (d, J=12.0Hz, 3H), 1.16-1.01 (m, 5H) , 0.96-0.84(m, 15H), 0.67(s, 3H), 0.58-0.44(m, 2H), 0.24(d, J=4.7Hz, 2H), 0.02--0.02(m, 5H).

[0694] Step 12: (5R)-1-cyclopropyl-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-1-ol (220-5) (36 mg, 0.068 mmol) was dissolved in THF (1 mL), and TBAF (1 mL, 1.000 mmol) was added dropwise at room temperature with stirring at room temperature. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1) until complete. The reaction solution was diluted with ethyl acetate (20 mL), washed with water (15 mL * 3), extracted with ethyl acetate (15 mL * 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by rapid column chromatography (petroleum ether:ethyl acetate = 90:10 to 75:25) to give a white solid 24-[cyclopropyl(hydroxy)methyl]cholan-3β-ol (220) (17.88 mg, 0.122 mmol, purity 94.86%, yield 89%). 1 H NMR (400MHz, Chloroform-d) δ3.63-3.54 (m, 1H), 2.85 (td, J=7.8, 4.9Hz, 1H), 1.96 (dt, J=12.6, 3.4Hz, 1H), 1.86-1.75 (m, 2H), 1.74-1.58 (m, 3H), 1 .49-1.19 (m, 14H), 1.03 (ddd, J=22.5, 17.2, 8.7Hz, 7H), 0.90 (t, J=7.6Hz , 5H), 0.80 (s, 3H), 0.65 (s, 4H), 0.57-0.44 (m, 2H), 0.24 (d, J=4.7Hz, 2H). 13 C NMR (101MHz, Chloroform-d) δ71.40, 64.92, 63.38, 56.51, 56.09, 54.40, 45.03, 42.62, 39.97, 38.29, 37.56, 36.91, 35. 76, 35.39, 32.09, 31.44, 28.75, 28.10, 24.22, 21.26, 19.64, 18.67, 18.01, 12.28, 11.99, 3.35, 2.27.LC-MS: [M-OH-H2O] + =381

[0695] Example 222

[0696] Preparation of compound 2223β,25-dihydroxycholest-6(5)-en-7-one

[0697]

[0698] Acetic acid-(3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-oxonyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (216-2) (20 mg, 0.044 mmol) was dissolved in a mixture of tetrahydrofuran (4 mL) and methanol (1 mL), and lithium hydroxide (1.5 mL, 1.5 mmol) was added at room temperature. The mixture was stirred at room temperature for 2 hours and monitored by TLC (dichloromethane:methanol = 20:1). After the reaction was completed, water and ethyl acetate were added for extraction, the organic phase was separated and dried, the crude product was concentrated, and purified by column chromatography (dichloromethane:methanol = 100:1 to 30:1) to obtain 3β,25-dihydroxycholest-6(5)-en-7-one (222) (12 mg, 0.029 mmol, 66.05%) as a white solid. 1 H NMR (399MHz, Chloroform-d) δ5.67 (s, 1H), 3.65 (s, 1H), 2.54-2.45 (m, 1H), 2.37 (t, J = 11.9Hz, 2H), 2.22 (t, J = 11.3Hz, 1H), 1.97 (dd, J = 33.6, 1 2.3Hz, 4H), 1.63 (d, J=15.0Hz, 2H), 1.51-1.21 (m, 13H), 1.19 (s, 6H), 1.18 (s, 3H), 1.07 (d, J=9.6Hz, 2H), 0.92 (d, J=6.4Hz, 3H), .0.66 (s, 3H). 13 CNMR (100MHz, Chloroform-d) δ202.24, 165.04, 126.09, 109.99, 71.09, 54.69, 49 .92, 49.88, 43.08, 38.67, 38.24, 36.30, 35.65, 31.16, 28.54, 21.18.LC-MS: [M+H] + =417.55.

[0699] Example 223

[0700] Preparation of compound 223 cholester-6(5)-ene-3β,7,25-triol

[0701]

[0702] In the first step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-oxonyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (216-2) (50 mg, 0.11 mmol) was dissolved in a mixture of tetrahydrofuran (5 mL) and methanol (2.5 mL). Sodium borohydride (6.19 mg, 0.16 mmol) was added at room temperature, and the mixture was stirred at room temperature for 1 hour. The mixture was monitored by TLC (petroleum ether:ethyl acetate = 2:1). After the reaction was completed, water and ethyl acetate were added for extraction. The organic phase was separated, dried, and the concentrated crude product, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-4-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (223-1) (36 mg, 0.074 mmol, 68.1%), was directly added to the next reaction step.

[0703] In the second step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-4-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (223-1) (36 mg, 0.08 mmol) was dissolved in a mixture of tetrahydrofuran (3 mL) and methanol (1.5 mL), and lithium hydroxide (0.5 mL, 0.5 mmol) was added. The mixture was stirred at room temperature for 1 hour and monitored by TLC (DCM:MeOH = 20:1). After the reaction was completed, water and ethyl acetate were added for extraction. The organic phase was separated and dried to obtain 40 mg of crude product. The crude product was purified by column chromatography (DCM:MeOH = 20:1) to obtain cholesterol-6(5)-ene-3β,7,25-triol (223) (18 mg, 0.041 mmol, 52.27%) as a white solid. 1HNMR (399MHz, Chloroform-d) δ5.27 (s, 1H), 3.84 (s, 1H), 3.58-3.49 (m, 1H), 2. 34-2.22 (m, 2H), 2.00 (d, J=11.0Hz, 1H), 1.84 (d, J=11.6Hz, 3H), 1.49 (s, 4H), 1. 40 (d, J=9.1Hz, 5H), 1.35 (s, 3H), 1.28 (d, J=7.4Hz, 3H), 1.20 (s, 6H), 1.16-1.07 (m, 3H), 1.03 (s, 3H), 0.92 (d, J = 6.5Hz, 3H), 0.85 (d, J = 7.1Hz, 1H), 0.68 (s, 3H). 13 C NMR (100MHz, CDCl3) δ143.46, 77.30, 76.98, 76.67, 71.10, 55.35, 48.22, 44.37, 42.9 2, 40.89, 39.52, 36.42, 35.67, 31.54, 29.66, 28.54, 26.34, 21.05.LCMS: [M-OH-H2O] + =383.5

[0704] Example 236

[0705] Preparation of compound 236(1R,3aR,6aR,8S,10aR,12aR)-1-((R)-6-hydroxy-6-methylheptane-2-yl)-7,7,10a,12a-tetramethylhexadecylhydro-4h-cyclopenta[1,2]phenanthrene[10,1-bc]furan-8-ol

[0706]

[0707]

[0708] In the first step, acetic acid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-4-formylbutyl-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecano-1H-cyclopenta[2,1-i]phenanthrene-7-yl ester (II) (500 mg, 1.1 mmol) was dissolved in tetrahydrofuran (30 mL), the mixture was purged with N2 three times, cooled to 0 °C, potassium tert-butoxide (253 mg, 2.3 mmol) solid was added, and the mixture was stirred at 0 °C for 30 min. Then, diethyl phosphonate (2-methoxy-2-oxoethyleneethyl) phosphonate (500 mg, 1.1 mmol) was dissolved in THF (5 mL) and added dropwise to the above reaction system. The mixture was allowed to rise naturally to room temperature and stirred for 2 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). The mixture was cooled to 0°C, and 2 mL of 1M / HCl was added. The mixture was separated, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and evaporated to dryness. The solution was then column-secreted to give a white solid (2E,6R)-6-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetyloxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecanoic acid methyl ester (67-1) (430 mg, 0.86 mmol, 76.65%). NMR (399MHz, CDCl3) δ7.03-6.85 (m, 1H), 5.80 (d, J=15.6Hz, 1H), 4.48 (dd, J=1 1.7, 4.5Hz, 1H), 3.77-3.62 (m, 3H), 2.25 (s, 1H), 2.12-1.92 (m, 8H), 1.89 (d, J =21.3Hz, 1H), 1.571.39 (m, 4H) 1.57-1.39 (m, 5H), 1.36-1.21 (m, 3H), 1.15 (dd , J=19.7, 10.5Hz, 3H), 0.98 (s, 3H), 0.87 (dd, J=14.6, 4.9Hz, 12H), 0.66 (s, 3H)

[0709] The second step involves weighing methyl(R)-6-((3S,5R,10S,13R,14R,17R)-3-acetoxy-4,4,10,13,14-pentylmethyl-2,3,4,4,5,6,7,10,11,12,13,14,15,16,17-tetradecanehydro-1H-cyclopentan[a]phenanthrene-17-yl)heptanoate (67-1) (100 mg, 0.2 mmol, 1.0 eq.), dissolving it in dichloromethane (10 mL), and then adding m-chloroperoxybenzoic acid (77.61 mg, 0.45 mmol, 2.3 eq.) under ice bath conditions. After the addition is complete, the mixture is brought back to room temperature and stirred continuously. TLC monitoring (petroleum ether:ethyl acetate = 10:1, Rf1 = 0.5, Rf2 = 0.3) was performed. After the reaction was completed, saturated sodium bicarbonate aqueous solution (5.0 mL) was added, and the mixture was extracted with dichloromethane (5.0 mL * 3). The organic phase was dried and concentrated to obtain methyl (6R)-6-((3S,5S,10S,13R,14R,17R)-3-acetoxy-4,4,10,13,14-pentamethyltetradecanehydro-11H-8,9-epoxycyclopentan[a]phenanthrene-17-yl)heptanoate (146-1) (89 mg, 0.17 mmol, 86%), a white crude solid. The unpurified product was added to the next step of the reaction.

[0710] In the third step, methyl (6R)-6-((3S,5S,10S,13R,14R,17R)-3-acetoxy-4,4,10,13,14-pentamethyltetradecanehydro-11H-8,9-epoxycyclopentan[a]phenanthrene-17-yl)heptanoate (146-1) (89 mg, 0.17 mmol) was weighed and dissolved in acetone (10 mL). Concentrated sulfuric acid (1 d) was added at room temperature, followed by stirring at room temperature for 5 hours. The mixture was monitored by TLC (petroleum ether:ethyl acetate = 10:1, Rf1 = 0.3, Rf2 = 0.6). After the reaction was completed, acetone was removed by concentration, saturated sodium bicarbonate aqueous solution (10.0 mL) was added, and then extracted with ethyl acetate (8.0 mL * 3). The organic phase was separated, dried, concentrated, and purified by column chromatography (petroleum ether: ethyl acetate = 30:1 ~ 10:1) to obtain methyl(R)-6-((3S,5R,10S,13R,14R,17R)-3-acetoxy-4,4,10,13,14-pentylmethyl-2,3,4,5,6,10,12,13,14,15,16,17-dodecylhydro-1H-cyclopentan[a]phenanthrene-17-yl)heptanoate (146-2) (81 mg, 0.16 mmol, 94%). 1H NMR(399MHz,CDCl3)δ5.43(s,1H),5.29(d,J=5.8Hz,1H),4.49(dd,J=11.1,4.8Hz,1H),3.65(s,3H),2.29(t,J=7.4Hz,2H),2.18(d,J=17.8Hz,1H),2.10-2.04(m,2H),2.03(s,4H),2.00-1.93(m,2H),1.76-1.68(m,2H),1.65-1.57(m,3H),1.50(d,J=4.6Hz,1H),1.42-1.34(m,4H),1.27(d,J=10.2Hz,2H),1.20-1.11(m,2H),1.071.00(m,1H),0.98(s,3H),0.93(s,3H),0.86(d,J=6.9Hz,9H),0.53(s,3H). 13 CNMR(101MHz,CDCl3)d 174.36,170.99,145.53,142.71,119.83,116.53,80.81,77.31,76.99,76.68,51.46,50.91,50.28,49.20,43.68,37.77,37.57,37.19,36.03,35.70,35.35,34.16,31.44,28.06,27.85,25.90,25.50,25.37,24.22,22.76,21.33,18.40,16.91,15.62,1.00.

[0711] In the fourth step, acetic acid-(1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-6-methoxy-6-oxoylide-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,5,5a,6,7,8,9,9a,11,11a-dodecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (146-2) (500 mg, 1.03 mmol) was weighed into a 100 mL single-necked flask, dissolved in acetic acid (10 mL), and then hydrogen peroxide (1.6 mL, 14.832 mmol) was added dropwise at room temperature. After the addition was complete, the mixture was stirred at room temperature, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1) to ensure complete reaction. After the reaction solution was lyophilized, the pH was adjusted to 7 with saturated sodium bicarbonate aqueous solution, ethyl acetate was added, the organic phase was separated, dried, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 50:1 to 20:1) to obtain 220 mg of crude product mixture. The crude product was slurried with ether (1 mL) and filtered to obtain acetic acid-(1R,3aS,5aR,7S,9aS,11aR)-1-[(2R)-6-methoxy-6-oxylidene-2-yl]-3a,6,6,9a,11a-pentamethyl-4-oxylidene-2,3,3a,3b,4,5,5a,6,7,8,9,9a,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (237-1) (44.9 mg, 0.09 mmol, 8.71%) as a white solid. 1 H NMR (399MHz, Chloroform-d) δ5.52-5.46 (m, 1H), 4.58 (dd, J=11.1, 4.9Hz, 1H), 3.65 (d, J=0.8Hz, 3H), 3.13 (s, 1 H), 3.02-2.90 (m, 1H), 2.40-2.33 (m, 1H), 2.30-2.20 (m, 3H), 2.05 (d, J=2.0Hz, 4H), 2.00 (d, J=3.5Hz, 2H), 1.89 (d, J=13.4Hz, 2H), 1.77-1.67 (m, 3H), 1.48 (t, J=5.2Hz, 1H), 1.46-1.34 (m, 4H), 1.241.15 (m, 1H), 1.09 (d, J=3. 8Hz, 4H), 1.05-1.00 (m, 1H), 0.92 (d, J=10.3Hz, 6H), 0.86 (d, J=6.0Hz, 3H), 0.81 (d, J=1.0Hz, 3H), 0.61 (s, 3H). 13C NMR (100MHz, CDCl3) δ209.12, 174.29, 170.87, 142.50, 121.71, 80.12, 77.33, 77.01, 76.70, 56. 19, 51.42, 50.46, 47.58, 47.15, 47.12, 46.62, 44.57, 42.49, 38.93, 38.54, 38.31, 37.96, 37.83, 37.34, 36.99, 35.83, 35.67, 35.61, 35.57, 35.33, 34.47, 34.34, 28.45, 28.34, 27.52, 27.25, 26.01, 24.77, 24.60, 23.88, 21.70, 21.23, 20.37, 18.36, 18.25, 17.71, 16.99, 16.71, 15.67, 15.14.

[0712] In the fifth step, acetic acid-(1R,3aS,5aR,7S,9aS,11aR)-1-[(2R)-6-methoxy-6-oxylidene-2-yl]-3a,6,6,9a,11a-pentamethyl-4-oxylidene-2,3,3a,3b,4,5,5a,6,7,8,9,9a,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (237-1) (91 mg, 0.182 mmol) was dissolved in a mixed solvent of methanol (2 mL) and tetrahydrofuran (4 mL). Sodium borohydride (6.88 mg, 0.182 mmol) was added at room temperature, and the mixture was stirred overnight. The reaction was detected by TLC (petroleum ether:ethyl acetate = 3:1) to confirm its completeness. Saturated sodium bicarbonate aqueous solution and ethyl acetate were added, the organic phase was separated, dried, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 10:1 to 5:1) to give acetic acid-(1R,3aS,5aR,7S,9aS,11aR)-4-hydroxy-1-[(2R)-6-methoxy-6-oxoylide-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,3b,4,5,5a,6,7,8,9,9a,11,11a-tetradecanohydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (237-2) (45 mg, 0.081 mmol, 44.33%) as a white solid. 1H NMR (399MHz, Chloroform-d) δ5.29 (d, J=5.9Hz, 1H), 4.46 (dd, J=11.5, 4.2Hz, 1H), 3.67 (s , 1H), 3.65 (s, 3H), 2.26 (q, J=8.2Hz, 2H), 2.16-2.07 (m, 1H), 2.03 (s, 4H), 1.93 (dd, J=19. 6, 8.4Hz, 3H), 1.82-1.59 (m, 6H), 1.49 (d, J=13.7Hz, 4H), 1.44-1.25 (m, 7H), 1.07 (s, 3H), 1.06-0.91 (m, 2H), 0.89 (t, J=3.3Hz, 6H), 0.85 (d, J=3.0Hz, 3H), 0.81 (s, 3H), 0.64 (s, 3H).

[0713] Step 6: Weigh acetic acid-(1R,3aS,5aR,7S,9aS,11aR)-4-hydroxy-1-[(2R)-6-methoxy-6-oxoylide-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,3b,4,5,5a,6,7,8,9,9a,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (237-2) (20 mg, 0.040 mmol) and dissolve it in a mixed solvent of ethyl acetate (3 mL) and acetic acid (0.3 mL). Add palladium on carbon (10% Pd), water containing 40-60% water (30 mg, 0.282 mmol), and platinum dioxide (3 mg, 0.013 mmol). Then, replace with hydrogen, raise the temperature to 80 °C, and raise the pressure to 200 psi with stirring. The reaction was monitored by HNMR. After stirring overnight (~48 hours * 3), heating was stopped, and the mixture was cooled to room temperature and filtered. The filtrate was added to a saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and the organic phase was separated. After drying and concentration, the product was given as acetic acid-(1R,3aS,5aR,7S,9aR,11aR)-4-hydroxy-1-[(2R)-6-methoxy-6-oxoylidene-2-yl]-3a,6,6,9a,11a-pentamethylhexadecyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (237-3) (15.6 mg, 0.029 mmol, 70%). 1H NMR (400MHz, Chloroform-d) δ4.50-4.40 (m, 1H), 3.65 (s, 3H), 3.59 (d, J=4.2Hz, 1H), 2.26 (q, J=8.4Hz, 2H), 2.03 (s, 3H), 1.90-1.82 (m, 1H), 1.63 (d, J=6.7Hz, 6H), 1.43 (s, 2H), 1.3 8(s, 2H), 1.35(s, 3H), 1.31(s, 1H), 1.26(s, 3H), 1.23(s, 3H), 1.09-1.02(m, 2H), 0.92(s, 3H), 0.88 (d, J=4.9Hz, 3H), 0.86 (d, J=1.2Hz, 6H), 0.84 (s, 3H), 0.81 (s, 1H), 0.75 (s, 3H). 13 C NMR (100MHz, Chloroform-d) δ174.33, 170.98, 110.00, 47.26, 46.18, 45.96, 37.61, 36.57, 35.73, 34.49, 31.71, 31.49, 23.97, 21.71.

[0714] In step seven, acetic acid-(1R,3aS,7S,9aR,11aR)-4-hydroxy-1-[(2R)-6-methoxy-6-oxylidene-2-yl]-3a,6,6,9a,11a-pentamethylhexadecyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (237-3) (70 mg, 0.139 mmol) was dissolved in benzene (5 mL), and high-lead acetate (1476.94 mg, 1.387 mmol) was added at room temperature. After nitrogen purging, the temperature was raised to 90 °C, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1) to ensure complete reaction. Cool to room temperature, add 10% potassium iodide solution. The reaction solution turns yellow and turbid. After stirring for 5 minutes, add saturated sodium thiosulfate solution until the system is colorless, clear and transparent. Then extract with ethyl acetate, separate the organic phase, dry and concentrate the crude product. Purify by column chromatography (petroleum ether:ethyl acetate = 20:1~10:1~5:1) to give methyl (5R)-5-((1R,3aR,6aR,8S,10aR,12aR)-8-acetoxy-7,7,10a,12a-tetramethylhexadecylhydro-4h-cyclopenta[1,2]phenanthrene[10,1-bc]furan-1-yl)hexanoate (236-1) (25 mg, 0.040 mmol, 28.62%) white solid.

[0715] 1H NMR (399MHz, Chloroform-d) δ4.47 (dd, J=11.6, 4.5Hz, 1H), 4.15 (s, 1H), 3.95 (d, J=7.5Hz, 1H), 3 .65 (s, 3H), 3.31 (d, J=7.5Hz, 1H), 2.26 (q, J=7.4Hz, 2H), 2.02 (s, 3H), 1.94 (s, 1H), 1.72 (td, J=23 .0, 19.8, 13.7Hz, 6H), 1.51 (d, J=11.6Hz, 3H), 1.37 (tt, J=13.4, 6.9Hz, 6H), 1.24 (s, 3H), 1.20-1 .12 (m, 1H), 1.060.96 (m, 2H), 0.88 (d, J = 6.4Hz, 3H), 0.84 (d, J = 7.1Hz, 6H), 0.80 (d, J = 6.7Hz, 6H).

[0716] In step 8, methyl (5R)-5-((1R,3aR,6aR,8S,10aR,12aR)-8-acetoxy-7,7,10a-12a-tetramethylhexadecylhydro-4h-cyclopenta[1,2]phenanthrene[10,1-bc]furan-1-yl)hexanoate (236-1) (25 mg, 0.050 mmol) was weighed and dissolved in THF (3 mL). The solution was cooled in an ice bath, and methyllithium (0.466 mL, 0.746 mmol) was added dropwise. After the addition was complete, the reaction was detected by TLC (dichloromethane:methanol = 10:1). After extraction, saturated sodium bicarbonate aqueous solution was added, followed by extraction with ethyl acetate. The organic phase was separated, dried, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 10:1~5:1~1:1) to obtain (1R,3aR,6aR,8S,10aR,12aR)-1-((R)-6-hydroxy-6-methylheptane-2-yl)-7,7,10a,12a-tetramethylhexadecylhydro-4h-cyclopenta[1,2]phenanthrene[10,1-bc]furan-8-ol (236) (9.2 mg, 0.017 mmol, 34.12%), a white solid. 1H NMR (400MHz, Methanol-d4) δ4.17 (d, J=3.1Hz, 1H), 4.01 (d, J=7.4Hz, 1H), 3.34 (d, J=7.4H z, 1H), 3.14 (dd, J=9.9, 6.3Hz, 1H), 2.07-1.96 (m, 1H), 1.93-1.78 (m, 3H), 1.60 (ddq, J=21. 7, 10.7, 6.3, 5.7Hz, 6H), 1.47 (dd, J=6.2, 2.9Hz, 2H), 1.37 (d, J=15.1Hz, 3H), 1.32-1.19 (m , 7H), 1.14 (s, 6H), 1.02 (q, J=8.8Hz, 3H), 0.91 (t, J=5.4Hz, 9H), 0.81 (s, 3H), 0.76 (s, 3H).

[0717] 13 C NMR (101MHz, Methanol-d4) δ78.20, 77.72, 73.61, 70.00, 58.46, 52.25, 49.17, 43.80, 43.77, 42.01, 38.36, 36.59, 36.33, 35.5 4,34.68,34.63,30.71,27.85,27.65,27.32,26.85,26.47,24.46,20.49,19.39,17.79,17.50,13.88,12.61.LCMS: [M-H2O-OH] + =425.45.

[0718] Example 258

[0719] Preparation of compound 258 24-[(2-fluorophenyl)(hydroxy)methyl]cholan-6(5)-ene-3β,4β-diol

[0720]

[0721]

[0722] The first step compound (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-1-(2-fluorophenyl)hex-1-ol (269-5) (100 mg, 0.17 mmol, 1.0 eq) was dissolved in chloroform (10 mL), and selenium dioxide (46.47 mg, 0.42 mmol, 2.5 eq) was added. Then, N-methylmorpholine (50.83 mg, 0.50 mmol, 3.0 eq) was added with stirring. The reaction system was heated to 75 °C and refluxed and stirred at this temperature for 60 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 3:1), and a highly polar substance was generated. The reaction was then quenched with 15 mL of water, extracted three times with 30 mL of ethyl acetate, dried over anhydrous sodium sulfate on the organic phase, filtered and concentrated, and the crude product was purified by rapid column chromatography (petroleum ether: ethyl acetate = 90:10 to 80:20) to obtain the product 3β-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-24-[(2-fluorophenyl)(hydroxy)methyl]-14α-methylcholan-6(5)-en-4β-ol (258-1) (30 mg, 0.034 mmol, yield 20.5%). 1 H NMR (400MHz, CDCl3) δ7.46 (t, J=7.6Hz, 1H), 7.22 (d, J=8.0Hz, 1H), 7.15 (t, J=7. 5Hz, 1H), 7.02 (m, 1H), 5.31 (d, J=5.3Hz, 1H), 4.95 (m, 1H), 3.48 (m, 1H), 2.22 (dt , J=13.2, 11.8Hz, 2H), 1.96 (d, J=14.1Hz, 2H), 1.75 (m, 5H), 1.37 (d, J=26.9Hz, 3 H), 1.14 (m, 9H), 0.99 (s, 5H), 0.90 (m, 13H), 0.66 (d, J=2.1Hz, 3H), 0.05 (s, 6H).

[0723] In the second step, compound 3β-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-24-[(2-fluorophenyl)(hydroxy)methyl]-14α-methylcholan-6(5)-en-4β-ol (258-1) (30 mg, 0.05 mmol) was dissolved in tetrahydrofuran (5 mL), and 1 mol / L tetrabutylammonium fluoride solution (0.5 mL) was added. The mixture was heated to 50 °C and stirred. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 3:1) until complete. The reaction solution was diluted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by rapid column chromatography (petroleum ether: ethyl acetate = 60:40) to obtain a white solid, which was then prepared by chirality to obtain a white solid 24-[(2-fluorophenyl)(hydroxy)methyl]cholan-6(5)-ene-3β,4β-diol (258) (6.4 mg, 0.0072 mmol, yield 8.27%). 1 H NMR (400MHz, CDCl3) δ7.50-7.41(m, 1H), 7.25-7.20(m, 1H), 7.17-7.11(m 1H), 7.05-6.98(m, 1H), 5.68(d, J=3.5Hz, 1H), 5.00(dt, J=11.5, 5.9Hz, 1H), 4.15(d , J=2.6Hz, 1H), 3.57 (d, J=11.4Hz, 1H), 2.00 (d, J=12.5Hz, 3H), 1.91 (d, J=12.4Hz, 1H ), 1.82 (d, J = 13.8Hz, 3H), 1.66 (d, J = 11.5Hz, 3H), 1.57 (d, J = 4.3Hz, 3H), 1.48-1.34 ( m, 8H), 1.18 (s, 3H), 1.08 (d, J = 9.7Hz, 3H), 0.93-0.89 (m, 4H), 0.67 (d, J = 3.5Hz, 3H). 13 C NMR (101MHz, CDCl3) δ168.72, 160.30, 142.76, 127.29, 127.18, 124.26, 117.60, 115.34, 72.50, 56.90, 56.06, 56.02, 50.18, 42.33, 39.68, 38.62, 38.44, 36.90, 36.00, 35.70, 28.21, 25.42, 24.24, 22.34, 22.24, 21.06, 20.54, 18.63, 14.09, 11.86.LCMS: [M+H-3OH] + =431

[0724] Example 259

[0725] Preparation of compound 259, 4-methylcholest-5(6)-ene-3β,4,25-triol

[0726]

[0727] Compound (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-6-oxoylide-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (203-1) (50 mg, 0.11 mmol, 1.0 eq) was dissolved in anhydrous tetrahydrofuran (10 mL). Under nitrogen protection, the mixture was cooled in an ice bath, and methyl lithium (0.681 mL, 1.09 mmol) was carefully added using a syringe. The mixture was then stirred at room temperature, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 1:1) until complete. The reaction was cooled to 0°C, quenched with ice water, extracted with ethyl acetate, washed with saturated brine of the organic phase, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by rapid column chromatography to obtain a white solid 4-methylcholest-5(6)-ene-3β,4,25-triol (259) (18 mg, 0.04 mmol, yield 35.2%). 1 H NMR (400MHz, DMSO) δ5.93 (d, J=29.5Hz, 1H), 4.34 (d, J=3.6Hz, 1H), 4.11 (s, 1 H), 4.02 (s, 1H), 3.18 (dd, J=7.7, 3.7Hz, 1H), 2.08-1.93 (m, 3H), 1.79 (s, 1H), 1.68-1.52(m, 5H), 1.32(d, J=14.7Hz, 11H), 1.24(s, 4H), 1.14(s, 3H), 1.05(s , 6H), 0.97 (s, 3H), 0.90 (d, J=6.4Hz, 3H), 0.84 (d, J=7.2Hz, 1H), 0.65 (s, 3H). 13 C NMR (101MHz, CDCl3) δ153.63, 124.27, 82.26, 80.41, 74.21, 61.95, 60.68, 55.61, 49.33, 46.95, 41.93, 41.35, 40. 44, 36.80, 35.77, 34.38, 34.19, 33.02, 32.16, 29.66, 28.98, 25.80, 25.49, 25.36, 23.64, 16.84.LCMS: [M+H-H2O] + =415

[0728] Example 260

[0729] Preparation of compound 260 ster-6(5)-ene-3β,4β,19,25-tetraol

[0730]

[0731] In the first step, methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (151-6) (1 g, 2.249 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL), and perchloric acid (0.727 mL, 9.00 mmol, 75%, 4.0 eq) and water (1 mL) were added. The reaction system was cooled to below 10 °C, and a tetrahydrofuran solution of N-bromosuccinimide (1.60 g, 9.00 mmol, 4.0 eq) was added. The reaction system was protected from light and stirred in an ice-water bath for 1.5 h. The reaction was then cooled to room temperature, and the completeness of the reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). The reaction was quenched with saturated sodium sulfite solution, and the product was extracted with water (15 mL) and ethyl acetate (10 mL × 3). The product was dried and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 90:10 to 88:12) to give a white solid (5R)-5-[(1R,3aS,3bS,5aR,5R,7S,9aR,9bS,11aR)-7-acetoxy-5a-bromo-5-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (260-1) (400 mg, purity 90%, yield 29.6%).

[0732] 1 H NMR (400MHz, CDCl3) δ5.47 (dt, J=16.2, 5.5Hz, 1H), 4.21-4.16 (m, 1H), 3.67 (s , 3H), 2.50 (dd, J=13.5, 10.5Hz, 1H), 2.35-2.16 (m, 4H), 2.03 (d, J=3.5Hz, 3H) , 2.01-1.94(m, 2H), 1.88-1.65(m, 6H), 1.60-1.53(m, 5H), 1.46-1.39(m, 2H), 1.30 (d, J=12.2Hz, 4H), 1.22-1.04 (m, 5H), 0.93 (d, J=6.5Hz, 3H), 0.67 (s, 3H).

[0733] In the second step, methyl (5R)-5-[(1R,3aS,3bS,5aR,5R,7S,9aR,9bS,11aR)-7-acetoxy-5a-bromo-5-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (260-1) (400 mg, 0.74 mmol, 1.0 eq) was dissolved in cyclohexane (12 mL) and benzene (3 mL). Diethyl iodophenyl ester (359.08 mg, 1.11 mmol, 1.5 eq) and iodine (224.95 mg, 0.89 mmol, 1.2 eq) were added. The reaction system was sonicated at 45 °C and 40 kHz for 1 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1) to ensure complete reaction. The reaction was quenched with saturated sodium sulfite (100 mL), extracted with ethyl acetate (50 mL × 3), the combined organic phases were washed with water (30 mL), dried, and concentrated to give the crude product. The crude product was subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 92:8) to give a white solid (5R)-5-[(1R,2S,5R,6R,9S,10S,12R,13R,15S)-15-acetoxy-13-bromo-5-methyl-19-oxapentane[10.5.2.01,13.05,9.02,10]nonadecan-6-yl]hexanoate methyl ester (260-2) (240 mg, purity 95%, yield 57.2%). 1 H NMR (400MHz, CDCl3) δ5.19 (dt, J=15.8, 5.4Hz, 1H), 4.06 (d, J=4.5Hz, 1H), 3.92 (d, J=8.4Hz, 1H), 3.74 (d, J=8.3Hz, 1H), 3.66 (s, 3H), 2.37-2.18 (m, 5H), 2.03 (s, 4H), 2 .01-1.95 (m, 3H), 1.89-1.77 (m, 2H), 1.65 (tdd, J=21.4, 10.5, 4.9Hz, 6H), 1.55-1.4 5(m, 8H), 1.41-1.36(m, 3H), 1.28-1.01(m, 9H), 0.91(d, J=6.6Hz, 3H), 0.69(s, 3H).

[0734] In the third step, the reactant (5R)-5-[(1R,2S,5R,6R,9S,10S,12R,13R,15S)-15-acetoxy-13-bromo-5-methyl-19-oxapentane[10.5.2.01,13.05,9.02,10]nonadecan-6-yl]hexanoate (260-2) (240 mg, 0.445 mmol, 1.0 eq) was dissolved in isopropanol (10 mL), and zinc powder (145.41 mg, 2.224 mmol, 5.0 eq) and acetic acid (0.356 mL, 6.227 mmol, 14 eq) were added. The reaction system was stirred at 90 °C for 3 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1) to ensure complete reaction. The zinc powder was removed by filtration, the organic phase was collected, and evaporated to dryness. The obtained oily substance was dissolved in ethyl acetate (10 mL), water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phase was collected to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 90:10 to 89:11) to give a white solid (5R)-5-[(1R,3aS,3bS,7S,9aS,9bS,11aR)-7-acetoxy-9a-(hydroxymethyl)-11a-methyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (260-3) (170 mg, purity 95%, yield 76.5%). 1 H NMR (400MHz, CDCl3) δ5.80-5.75 (m, 1H), 4.70-4.59 (m, 1H), 3.83 (d, J=11.5Hz, 1H), 3.66 (s, 3H) , 3.62 (d, J=11.4Hz, 1H), 2.42 (ddd, J=12.9, 4.8, 2.1Hz, 1H), 2.33-2.21 (m, 3H), 2.03 (s, 3H), 1. 95(dt, J=13.8, 3.5Hz, 1H), 1.87-1.76(m, 3H), 1.73-1.43(m, 9H), 1.42-1.32(m, 3H), 1.26(dd, J =14.5, 7.1Hz, 3H), 1.18-1.02 (m, 5H), 0.93 (d, J = 6.6Hz, 3H), 0.87 (d, J = 6.1Hz, 1H), 0.73 (s, 3H).

[0735] In the fourth step, compound (5R)-5-[(1R,3aS,3bS,7S,9aS,9bS,11aR)-7-acetoxy-9a-(hydroxymethyl)-11a-methyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (260-3) (100 mg, 0.217 mmol, 1.0 eq) was dissolved in chloroform (10 mL), and selenium dioxide (IV) (36.13 mg, 0.326 mmol) and N-methylmorpholine-N-oxide (76.29 mg, 0.651 mmol) were added. The reaction mixture was stirred at 80 °C for 18 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 1:1) to ensure complete reaction. Water (30 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with water (30 mL), dried, and concentrated to obtain the crude product. The crude product was subjected to silica gel column chromatography (petroleum ether:ethyl acetate = 68:32) to obtain a white solid (5R)-5-[(1R,3aS,3bS,6R,7S,9aS,9bS,11aR)-7-acetoxy-6-hydroxy-9a-(hydroxymethyl)-11a-methyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (260-4) (32 mg, purity 90%, yield 27.8%). 1 H NMR (400MHz, CDCl3) 65.98 (dd, J=4.5, 2.5Hz, 1H), 4.80-4.74 (m, 1H), 4.26 (d, J=2.5Hz, 1 H), 3.83 (d, J=11.2Hz, 1H), 3.70 (d, J=11.2Hz, 1H), 3.66 (s, 3H), 2.36-2.13 (m, 4H), 2.10 (s, 3H), 2.07-1.78 (m, 10H), 1.73-1.66 (m, 2H), 1.63-1.47 (m, 6H), 1.44-1.35 (m, 3H), 1. 29-1.22 (m, 4H), 1.11 (tdd, J=18.7, 12.5, 6.7Hz, 6H), 0.93 (d, J=6.5Hz, 3H), 0.71 (s, 3H).

[0736] In the fifth step, the reactant (5R)-5-[(1R,3aS,3bS,6R,7S,9aS,9bS,11aR)-7-acetoxy-6-hydroxy-9a-(hydroxymethyl)-11a-methyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (260-4) (30 mg, 0.063 mmol) was dissolved in tetrahydrofuran (10 mL). After cooling the reaction system to 0 °C, methyl magnesium chloride (0.21 mL, 0.63 mmol, 3 mol / L) was slowly added. The reaction system was stirred at room temperature for 18 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 1:2). 20 mL of water was added to the reaction system, and the mixture was extracted with ethyl acetate (25 mL × 3). The organic phase was collected, dried with anhydrous sodium sulfate, and then evaporated under vacuum to obtain the crude product. The crude product was dissolved in ethyl acetate and subjected to column chromatography (petroleum ether: ethyl acetate = 36: 64) to give cholesterol-6(5)-ene-3β,4β,19,25-tetraol (260) (8.62 mg, purity 92.8%, yield 29.2%). 1 H NMR (400MHz, DMSO) δ5.69 (d, J=2.3Hz, 1H), 4.61 (d, J=2.7Hz, 1H), 4.33 (d, J=6.5Hz, 1H), 4.02 (s, 1H), 3.99-3.94 (m, 1H), 3.92 (s, 1H), 3.65 (ddd, J=17.7, 11.0, 5.8Hz, 2H), 3.29 (s , 1H), 2.05-1.83(m, 4H), 1.82-1.68(m, 3H), 1.64-1.44(m, 4H), 1.401.28(m, 6H), 1.26-1. 16 (m, 6H), 1.05 (s, 6H), 0.89 (d, J=6.5Hz, 3H), 0.80 (dd, J=13.6, 5.3Hz, 2H), 0.67 (s, 3H). 13 C NMR (101MHz, CDCl3) δ145.53, 134.05, 81.78, 76.90, 73.96, 68.91, 62.34, 60.77, 55.60, 49.36, 47.24, 45.88 , 41.42, 40.45, 38.27, 37.30, 36.56, 36.37, 34.65, 34.45, 30.88, 29.05, 26.22, 25.69, 25.45, 23.79, 17.10.

[0737] Example 269

[0738] Preparation of compound 26924-[(2-fluorophenyl)(hydroxy)methyl]cholan-6(5)-en-3β-ol

[0739]

[0740]

[0741] The first step involved dissolving methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (151-6) (2 g, 4.50 mmol) in methanol (50 mL), adding concentrated sulfuric acid (98%) (1 mL, 18.76 mmol) dropwise, and then heating to reflux with stirring for 1 hour. After the reaction was complete as monitored by TLC (petroleum ether:ethyl acetate = 10:1), most of the solvent was concentrated off the reaction solution. Ethyl acetate (~100 mL) was added, and the solution was washed twice with saturated sodium bicarbonate solution (~50 mL x 2), and once with saturated brine (~50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. The crude product was purified by rapid column chromatography (petroleum ether:ethyl acetate = 90:10 to...). (80:20) gave a white solid (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (269-1) (1.8 g, 4.02 mmol, yield 89.5%). 1 H NMR (400MHz, CDCl3) δ5.37-5.33 (m, 1H), 3.67 (s, 3H), 3.52 (dd, J=7.7, 3.3Hz, 1H), 2.35-2.20 (m, 4H), 2.03-1.93 (m, 2H), 1.84 (ddd, J= 12.5, 7.5, 4.3Hz, 3H), 1.75-1.60 (m, 4H), 1.59-1.35 (m, 9H), 1.33-1.04 (m, 7H), 1.03-0.99 (m, 3H), 0.93 (d, J=6.6Hz, 3H), 0.68 (s, 3H).

[0742] In the second step, compound (5R)-5-[(1R,3aS3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (269-1) (1.7 g, 4.38 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (50 mL), followed by the sequential addition of tert-butyldimethylchlorosilane (3.3 g, 21.85 mmol, 5.0 eq) and imidazole (1.5 g, 21.85 mmol, 5.0 eq). The reaction was heated to 100 °C, and TLC (petroleum ether:ethyl acetate) was performed. After the reaction was completed (10:1), the reaction solution was cooled to room temperature, ethyl acetate (150 mL) was added, the organic phase was washed three times with water, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by rapid silica gel column chromatography (petroleum ether:ethyl acetate = 90:10) to give a colorless waxy substance (5R)-5-[(1R,3aS,3bR,7S,9bS,11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-11a-methyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (269-2) (1.9 g, 3.40 mmol, yield 77.7%). 1 H NMR (400MHz, CDCl3) δ5.26-5.18 (m, 1H), 3.57 (s, 3H), 3.43-3.33 (m, 1H), 2.16 (d, J=1.9Hz, 4H), 1.93-1.82 (m, 2H), 1.65-1.55 (m, 2H), 1.39 (d, J=3.0Hz, 9H), 0.99 (s, 7H), 0.90 (s, 5H), 0.85-0.79 (m, 21H), 0.57 (s, 3H), -0.00 (s, 6H).

[0743] The third step compound is (5R)-5-[(1R,3aS,3bR,7S,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-11a-methyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (269-2) (1.8 g, 3.5 g). 8 mmol (1.0 eq) was dissolved in tetrahydrofuran (50 mL), and lithium aluminum hydride (0.14 g, 3.580 mmol, 1.0 eq) was added. The mixture was stirred at room temperature for 30 min, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was complete, the reaction solution was quenched with sodium sulfate decahydrate, then filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by rapid silica gel column chromatography (petroleum ether:ethyl acetate = 80:20). (to 70:30) yielded a white solid (5R)-5-[(1R,3aS,3bR,7S,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-11a-methyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-1-ol (269-3) (1.7 g, 3.22 mmol, 90.0% yield). 1 H NMR (400MHz, CDCl3) δ5.26 (d, J=5.3Hz, 1H), 3.59 (t, J=6.6Hz, 2H), 3.47-3.37 (m, 1H), 2.27-2 .17(m, 1H), 2.11(ddd, J=13.4,5.0, 2.2Hz, 1H), 1.93(s, 2H), 1.77(s, 2H), 1.66(d, J=12.2Hz, 1H), 1.47(ddd, J=13.9, 8.7, 4.5Hz, 7H), 1.37-1.29(m, 3H), 1.25-1.15(m, 2H), 1.13-0.97(m, 5H), 0.94 (s, 3H), 0.87 (d, J=6.3Hz, 4H), 0.84-0.81 (m, 10H), 0.62 (s, 3H), 0.02--0.02 (m, 6H).

[0744] In the fourth step, compound (5R)-5-[(1R,3aS,3bR,7S,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-11a-methyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-l-ol (269-3) (800 mg, 1.69 mmol, 1.0 eq) was dissolved in dichloromethane (40 mL), and Dysmartin oxidant (833 mg, 1.96 mmol, 1.2 eq) was added. The mixture was stirred at room temperature. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, the reaction solution was quenched with saturated sodium sulfite solution, washed with saturated sodium bicarbonate, dried with anhydrous sodium sulfate, filtered, concentrated, and the crude product was obtained. The crude product was purified by rapid column chromatography (petroleum ether: ethyl acetate = 90:10) to give a white solid (5R)-5-[(1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-7-{[dimethyl(2-methylpropyl-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b, 4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanal (269-4) (500 mg, 0.924 mmol, yield 56.48%). 1 H NMR (400MHz, CDCl3) δ9.71 (t, J=1.8Hz, 1H), 5.26 (d, J=5.2Hz, 1H), 3.48-3.36 (m, 1H), 2.34 (d, J=6.7Hz, 3H), 2.11 (ddd, J=13.3, 5.0, 2.1Hz, 1H), 1.93 (s, 2H), 1.75 (d, J=13. 3Hz, 2H), 1.66 (dd, J=12.7, 4.6Hz, 2H), 1.47-1.28 (m, 8H), 1.20 (s, 2H), 1.15-0.97 (m, 6H), 0.94 (s, 4H), 0.89 (d, J=6.5Hz, 4H), 0.85-0.82 (m, 10H), 0.62 (s, 3H), 0.01 (m, 6H).

[0745] In the fifth step, compound 2-bromo-1-fluorobenzene (500 mg, 2.86 mmol) was dissolved in tetrahydrofuran (5 mL), purged with nitrogen three times, and the system was cooled to -78 °C in a dry ice bath. Then, 2.5 M n-butyllithium solution (1.26 mL, 3.14 mmol) was added dropwise using a syringe, and the reaction was maintained at this temperature for 1 hour. The reaction solution was then directly added to the next step.

[0746] The solid compound (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanal (269-4) (500 mg, 1.03 mmol) was dissolved in anhydrous tetrahydrofuran (20 mL) under nitrogen protection and cooled to -10 °C in an ice-salt bath. The prepared (2-fluorophenyl)lithium (6.26 mL) was slowly added, and the mixture was stirred at room temperature for 30 min. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 5:1). After the reaction was completed, the reaction solution was quenched with ice water, extracted twice with ethyl acetate, washed with saturated brine of the organic phase, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 90:10) to give a white solid (5R)-5-[(1R, 3aS, 3bS) ,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-1-(2-fluorophenyl)hex-1-ol (269-5) (450 mg, 0.58 mmol, yield 56.37%). 1 H NMR (400MHz, CDCl3) δ7.46 (t, J=7.5Hz, 1H), 7.26-7.19 (m, 1H), 7.15 (t, J=7.5Hz, 1H), 7.06-6. 98 (m, 1H), 5.35 (d, J=5.0Hz, 1H), 5.00 (dd, J=13.2, 6.1Hz, 1H), 3.52 (dt, J=15.8, 5.6Hz, 1H), 2 .34-2.18(m, 2H), 1.98(t, J=12.9Hz, 2H), 1.88-1.66(m, 5H), 1.60-1.31(m, 10H), 1.31-1.13(m , 3H), 1.13-1.03 (m, 4H), 1.00 (s, 3H), 0.96-0.87 (m, 16H), 0.66 (t, J=6.2Hz, 3H).0.01 (m, 6H).

[0747] The sixth-step compound was (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-1-(2-fluorophenyl)hex-1-ol (269-5) (250 mg, 0.43 mmol). Dissolved in tetrahydrofuran (10 mL), 1 mol / L tetrabutylammonium fluoride solution (1 mL) was added, the reaction was heated to 50 °C, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 5:1). After the reaction was completed, the reaction was cooled to room temperature, diluted with ethyl acetate, washed three times with saturated brine, dried with anhydrous sodium sulfate in the organic phase, filtered, concentrated to dryness to obtain crude product, and purified by rapid silica gel column chromatography (petroleum ether: ethyl acetate = 80:20) to obtain a white solid. Then, it was purified by chiral preparation (Column: Daicel CHIRALCEL IE, 250 mm / 30 mm I.D., 10 μm, Mobile phase: CO2 / MeOH [0.2% NH3 (7M Solution in MeOH)] = 60 / 40) to obtain a white solid 24-[(2-fluorophenyl)(hydroxy)methyl]cholan-6(5)-en-3β-ol (269) (50 mg, 0.09 mmol, yield 21.43%). 1 H NMR (400MHz, CDCl3) δ7.46 (t, J=7.5Hz, 1H), 7.26-7.19 (m, 1H), 7.15 (t, J=7.5Hz, 1H), 7.0 6-6.98 (m, 1H), 5.35 (d, J=5.0Hz, 1H), 5.00 (dd, J=13.2, 6.1Hz, 1H), 3.52 (dt, J=15.8, 5.6H z, 1H), 2.34-2.18 (m, 2H), 1.98 (t, J=12.9Hz, 2H), 1.88-1.66 (m, 5H), 1.60-1.31 (m, 10H), 1 .31-1.13(m, 3H), 1.131.03(m, 4H), 1.00(s, 3H), 0.96-0.87(m, 4H), 0.66(t, J=6.2Hz, 3H). 13CNMR(101 MHz, CDCl3) δ140.76, 131.84, 128.72, 127.25, 124.28, 121.73, 115.41, 115.15, 71.86, 68.67, 68.44, 68.38, 56.75, 56.06, 50.11, 42.33, 39.77, 38.63, 38.46, 37.25, 36.50, 35.72, 35.59, 31.90, 31.66, 28.24, 24.28, 22.37, 22.26, 21.08, 19.41, 18.61, 11.86.LCMS[M+H-2OH] + =433

[0748] Example 270

[0749] Preparation of compound 270N-[(3R)-3-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]butyl]-N-methyl-1,3-thiazacyclopentaman-2-carboxamide

[0750]

[0751]

[0752] In the first step, in a 50 mL round-bottom flask, 9a,11a-dimethyl-1-[(2R)-4-(methylamino)-2-yl]-2,3,3a,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetratetrahydro-1H-cyclopentane[1,2-i]phenanthrene-7-acetate (40 mg, 0.100 mmol) was dissolved in dichloromethane (51-4) (2 mL). Triethylamine (30.23 mg, 0.299 mmol) and 1,3-thiazolyl-2-carboxyl chloride (29.40 mg, 0.199 mmol) were added at room temperature. The mixture was stirred at room temperature for 3 hours. The reaction was monitored for completeness by TLC (petroleum ether / ethyl acetate = 4 / 1). After the reaction was complete, water (20 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (15 mL × 3). The extracted organic solution was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1). The solvent was removed by concentration under vacuum to give a white solid acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-9a,11a-dimethyl-1-[(2R)-4-[methyl(1,3-thiazacyclopentanylcarbonyl)amino]but-2-yl]-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (270-1) (29 mg, purity: 90%, yield: 51.11%). LCMS: [M+H + =513.3.

[0753] The second step involves preparing acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-9a,11a-dimethyl-1-[(2R)-4-[methyl(1,3-thiazacyclopentanylcarbonyl)amino]but-2-yl]-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (270-1)(31) at room temperature. Lithium hydroxide (mg, 0.060 mmol) was dissolved in a 50 mL round-bottom flask containing 1 mL of tetrahydrofuran and 1 mL of methanol. Lithium hydroxide monohydrate (12.68 mg, 0.302 mmol) and water (0.6 mL) were added at room temperature. The mixture was then stirred at room temperature for 2 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 2:1). After the reaction was complete, the mixture was quenched with 20 mL of water, extracted with 10 mL × 3 ethyl acetate solutions, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1). Ether:ethyl acetate = 2:1) yielded a white solid N-[(3R)-3-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]butyl]-N-methyl-1,3-thiazacyclopentaman-2-carboxamide (270) (17.59 mg, purity: 94.38%, yield: 58.33%).

[0754] 1 H NMR (400MHz, CDCl3) δ7.89-7.86 (m, 1H), 7.51 (t, J=2.7Hz, 1H), 5.35 (d, J=5.0 Hz, 1H), 4.10-3.97(m, 1H), 3.57-3.49(m, 3.5H), 3.11(s, 1.5H), 2.32-2.20(m, 2H), 2.02-1.95(m, 2H), 1.85-1.78(m, 3H), 1.62-1.41(m, 9H), 1.30-1.22(m, 3H ), 1.151.03 (m, 5H), 1.01 (s, 3H), 0.94 (d, J = 6.0Hz, 3H), 0.68 (d, J = 8.2Hz, 3H).

[0755] 13CNMR (101MHz, CDCl3) δ165.97, 165.16, 160.76, 160.21, 143.13, 140.77, 123.81, 123.64 ,121.67,121.64,71.79,56.72,55.81,55.77,50.07,48.57,47.53,42.41,42.38,42.29 ,39.74,39.69,37.25,36.77,36.50,35.25,34.52,34.10,33.99,32.71,31.88,31.65,2 9.71, 29.33, 28.26, 28.18, 24.27, 21.06, 19.40, 18.89, 18.79, 11.84, 11.79.LCMS[M+H] + =471.3

[0756] Examples 276 & 277

[0757] Preparation of compound 276 3β,5,25-trihydroxycholest-4-one

[0758]

[0759] Preparation of compound 277 3β,25-dihydroxycholest-6(5)-en-4-one

[0760]

[0761] In the first step, in a 50 mL round-bottom flask, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-5a,6-dihydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (248-5) (42 mg, 0.088 mmol) was dissolved in dichloromethane (1.5 mL). N-methylmorpholine oxide (15 mg, 0.13 mmol), 4A molecular sieve (21 mg), and tetrapropylammonium perruthenate (6 mg, 0.018 mmol) were added at room temperature. The mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 3:1). After the reaction was complete, saturated sodium sulfite aqueous solution (15 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (10 mL × 3). The extracted organic solution was washed with saturated brine (10 mL), dried with anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1) to obtain a white solid acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-5a-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-6-oxoylidehexadecyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (276-1) and acetic acid-(1R,3aS) A mixture of (2R)-6-hydroxy-6-methylhept-2-yl)-9a,11a-dimethyl-6-oxoylide-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanohydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (276-2) (35 mg, purity: 90%, yield: 76.90%).

[0762] LC-MS(276-1): [M+Na]+=499.4

[0763] LC-MS (276-2): [M+Na] + =481.4

[0764] In the second step, in a 50 mL round-bottom flask, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-5a-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-6-oxoylidenehexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (276-1) and acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a A mixture (276-2) of dimethyl-6-oxoylide-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl esters (35 mg, 0.074 mmol) was dissolved in tetrahydrofuran (1 mL) and methanol (1 mL). Potassium carbonate (51 mg, 0.37 mmol) and water (0.5 mL) were added at room temperature, followed by stirring at room temperature for 3 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 2:1). After the reaction was complete, the mixture was diluted with water (15 mL), extracted with ethyl acetate (10 mL × 3), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified and separated by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give a white solid 3β,5,25-trihydroxycholest-4-one (276) (11.05 mg, purity: 93.95%, yield: 54.38%) and 3β,25-dihydroxycholest-6(5)-en-4-one (277) (6.25 mg, purity: 100%, yield: 48.98%).

[0765] 276: 1 H NMR (400MHz, DMSO) δ4.21 (d, J=6.1Hz, 1H), 4.12-4.11 (m, 2H), 4.03 (s, 1H), 2.17-2.12 (m, 1H), 2.02-1.93 (m, 2H), 1.8 5-1.69 (m, 4H), 1.60-1.41 (m, 6H), 1.37-1.23 (m, 14H), 1.15 (s, 3H), 1.05 (s, 6H), 0.89 (d, J=6.4Hz, 3H), 0.65 (s, 3H). 13CNMR (101MHz, DMSO)211.02, 80.19, 77.97, 69.24, 56.28, 45.52, 44.58, 42.62, 36.65, 36.22, 35.74, 34. 53, 33.17, 29.88, 29.70, 28.51, 28.36, 26.27, 24.31, 21.09, 20.81, 18.99, 15.50, 12.35.LC-MS-MS[M+H] + =435

[0766] 277: 1 H NMR (400MHz, DMSO) δ7.71 (s, 1H), 4.03 (s, 1H), 3.18 (s, 0.5H), 3.02 (s, 0.5H), 2.93-2.85 (m, 1H), 2.33-2.26 (m, 1H), 2.12-1.69 (m, 9H), 160-1.27 (m, 15H), 1.12 (s, 3H), 1.05 (s, 6H), 0.90 (d, J=6.4Hz, 3H), 0.68 (s, 3H). 13 C NMR(101 MHz, DMSO) δ193.37, 141.85, 139.73, 69.23, 56.09, 56.00, 54.31, 44.60, 42.39, 37.73, 36.62, 35.66, 35.14 , 34.78, 32.79, 31.33, 29.88, 29.70, 28.26, 24.27, 22.79, 21.02, 20.69, 18.99, 17.26, 12.25.LC-MS[M+Na] + =439.3

[0767] Example 311

[0768] Preparation of compound 311 24-[hydroxy(2-methoxyphenyl)methyl]cholan-6(5)-en-3β-ol

[0769]

[0770] Referring to Example 269, o-fluorobromobenzene was replaced with o-methoxybromobenzene to obtain compound 24-[hydroxy(2-methoxyphenyl)methyl]cholan-6(5)-en-3β-ol(311)en-3β-ol (40 mg, purity: 98.78%, yield 78.38%). 1HNMR (400MHz, CDCl3) δ7.32-7.27 (m, 1H), 7.23 (d, J=7.8Hz, 1H), 6.95 (t, J=7.4Hz, 1H), 6.88 ( d, J=8.2Hz, 1H), 5.35 (d, J=5.1Hz, 1H), 4.85 (dt, J=9.5, 5.9Hz, 1H), 3.86 (s, 3H), 3.63-3.44 ( m, 1H), 2.35-2.18 (m, 2H), 1.98 (t, J=11.7Hz, 2H), 1.84 (d, J=10.0Hz, 5H), 1.77-1.71 (m, 1H), 1.61-1.33 (m, 9H), 1.30-1.03 (m, 7H), 1.00 (s, 3H), 0.95-0.87 (m, 4H), 0.67 (d, J=2.5Hz, 3H). 13 C NMR(101 MHz, CDCl3) δ156.59, 140.78, 132.76, 128.20, 127.05, 121.72, 120.75, 110.56, 71.82, 71.39, 70.98, 56.77, 56.15, 56.11, 55 .28, 50.12, 42.34, 42.31, 39.74, 37.83, 37.25, 35.88, 35.70, 31.90, 31.67, 28.25, 24.29, 22.63, 21.08, 19.40, 18.64, 11.66.

[0771] Example 314

[0772] Preparation of compound 314 24-[(4-fluorophenyl)(hydroxy)methyl]cholan-5(6)-ene-3β,4β-diol

[0773]

[0774]

[0775] The first step reaction involved dissolving methyl (5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (151-7) (1 g, 2.17 mmol, 1.0 eq) in methanol (50 mL), and adding potassium carbonate (3.00 g, 21.71 mmol, 10 eq). The reaction was allowed to proceed for 20 min, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 1:1). The reaction mixture was diluted with 10 mL of water, extracted with ethyl acetate (30 mL x 3), and the organic phase was collected, washed with 30 mL of water, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 73:27) to give a white solid (5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-dihydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (271-1) (760 mg, 1.725 mmol, purity: 95%, yield: 79%).

[0776] 1 H NMR (400MHz, CDCl3) δ5.71-5.63 (m, 1H), 4.14 (d, J=3.2Hz, 1H), 3.67 (s, 3H), 3 .56(dt, J=11.7, 4.1Hz, 1H), 2.36-2.19(m, 2H), 2.14-1.79(m, 6H), 1.76-1.62 (m, 6H), 1.59-1.51 (m, 3H), 1.49-1.33 (m, 5H), 1.25 (d, J=7.2Hz, 1H), 1.18 (s, 3H), 1.08 (ddd, J=17.0, 12.6, 7.8Hz, 5H), 0.93 (d, J=6.5Hz, 3H), 0.68 (s, 3H).

[0777] In the second step, methyl (5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-dihydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (271-1) (1.8 g, 4.30 mmol, 1.0 eq) was dissolved in acetone (50 mL), and p-toluenesulfonic acid (0.57 g, 3.01 mmol, 0.7 eq) and 4A molecular sieve were added. The resulting mixture was stirred at 25 °C for 1 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1) until complete. Water (10 mL) was added to the reaction system, and the aqueous layer was extracted with ethyl acetate (3 × 50 mL). The organic phases were combined and washed with saturated brine (50 mL). The mixture was dried over Na2SO4, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 90:10) to give a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecanoic acid methyl ester (271-2) (1.6 g, 3.14 mmol, purity: 90%, yield: 73%). 1 H NMR (400MHz, CDCl3) δ7.33 (d, J=7.3Hz, 1H), 7.23 (s, 1H), 7.13 (d, J=7.5Hz, 1H), 7. 03 (m, 1H), 6.02 (d, J = 5.0Hz, 1H), 5.68 (d, J = 3.4Hz, 1H), 4.14 (s, 1H), 3.58 (s, 1H), 2 .09(t, J=3.9Hz, 3H), 2.00(m, 2H), 1.78(m, 12H), 1.56(m, 4H), 1.38-1.30(m, 9H), 1 .18 (s, 3H), 1.00-0.95 (m, 7H), 0.88 (dd, J=13.7, 7.3Hz, 4H), 0.66 (d, J=3.4Hz, 3H).

[0778] In the third step, compound (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecanoic acid methyl 3aH-cyclopenta[1′,2′∶1,2]phenanthro[7,8-d][1,3]dioxane-8-yl]hexanoate (271-2) (1 g, 2.18 mmol, 1.0 eq) was dissolved in tetrahydrofuran (50 mL), purged with nitrogen, and then lithium aluminum hydride (0.12 g, 3.270 mmol, 1.5 eq) was added. The mixture was stirred at 25 °C for 1 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1) to ensure complete reaction. The product was quenched with sodium sulfate decahydrate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 80:20) to give a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecano-3aH-cyclopentazo[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxane-8-yl]hex-1-ol (271-3) (1000 mg, 2.09 mmol, purity: 90%, yield: 96%).

[0779] 1 H NMR (400MHz, CDCl3) δ5.80 (d, J=2.6Hz, 1H), 4.41 (d, J=5.8Hz, 1H), 4.09 (m, 1H), 3.64 (t, J=6.5Hz, 2H), 2.08 (m, 2H), 1.84 (tt, J=18.5, 7.8Hz, 2H ), 1.67(m, 8H), 1.41(td, J=12.7, 6.2Hz, 4H), 1.35(s, 3H), 1.24(m, 2H), 1.17 (s, 4H), 1.08 (m, 5H), 0.93 (d, J = 6.5Hz, 4H), 0.69 (d, J = 4.6Hz, 3H).

[0780] In the fourth step, compound (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecano-3aH-cyclopenta[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxane-8-yl]hex-1-ol (271-3) (1 g, 2.32 mmol, 1.0 eq) was dissolved in dichloromethane (50 mL), and Dysmartin oxidant (1.18 g, 2.79 mmol, 1.2 eq) was added. The mixture was stirred at 25 °C for 1 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1) to ensure complete reaction. The reaction mixture was quenched with saturated sodium sulfite, and water (10 mL) was added to the reaction system. The aqueous layer was extracted with dichloromethane (3 × 50 mL). The organic phases were combined and washed with saturated brine (10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 90:10) to give a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecano-3aH-cyclopentazo[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxane-8-yl]hexanal (271-4) (750 mg, 1.58 mmol, purity: 90%, yield: 68%).

[0781] 1 H NMR (400MHz, CDCl3) δ9.76 (s, 1H), 5.80 (d, J = 2.7Hz, 1H), 4.41 (d, J = 5.8Hz, 1H ), 4.10(dd, J=13.6, 6.3Hz, 1H), 2.39(m, 2H), 2.12(m, 1H), 2.01(m, 1H), 1.83(m , 1H), 1.66 (m, 10H), 1.53 (s, 4H), 1.42 (dd, J=12.5, 4.2Hz, 3H), 1.35 (s, 3H), 1 .17 (s, 3H), 1.10 (m, 4H), 0.92 (dd, J=18.2, 5.8Hz, 4H), 0.71 (d, J=12.1Hz, 3H).

[0782] Step 5: Weigh (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecano-3aH-cyclopenta[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxane-8-yl]hexanal (271-4) (100 mg, 0.23 mmol) and dissolve it in tetrahydrofuran (10 mL). Add p-fluorophenyl magnesium bromide (1.0 M, 0.6 mL) dropwise under ice bath conditions. After the addition is complete, bring the temperature to room temperature and stir. Detect by TLC (petroleum ether:ethyl acetate = 3:1). Phosphomolybdic acid is used for color development. After the reaction was completed, saturated ammonium chloride aqueous solution and ethyl acetate were added for extraction. The organic phase was separated, dried, concentrated, and purified by column chromatography (petroleum ether: ethyl acetate = 0-30%) to obtain (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecanohydro-3aH-cyclopenta[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxacyclopenta-8-yl]-1-(4-fluorophenyl)hex-1-ol (314-1) (98 mg, 0.19 mmol, 82.6%), a white solid. 1 H NMR (399MHz, Chloroform-d) δ7.29 (dd, J=8.4, 5.5Hz, 2H), 7.01 (t, J=8.6Hz, 2H), 5.78 (dd, J=4.8, 2.0Hz, 1H), 4.68-4.60 (m, 1 H), 4.39 (d, J=5.8Hz, 1H), 4.08 (q, J=6.6Hz, 1H), 2.10 (dd, J=12.6, 4.5Hz, 1H), 1.98 (d, J=12.5Hz, 1H), 1.78 (dt, J=14.6, 7. 4Hz, 2H), 1.70-1.61 (m, 4H), 1.56 (d, J=9.8Hz, 4H), 1.51 (s, 3H), 1.48 (s, 1H), 1.39 (d, J=13.6Hz, 3H), 1.33 (s, 3H), 1.25 (d, J=11.3Hz, 2H), 1.14 (s, 3H), 1.08 (dd, J=18.2, 9.7Hz, 4H), 1.03-0.90 (m, 3H), 0.87 (t, J=5.9Hz, 3H), 0.66 (d, J=2.6Hz, 3H).

[0783] Step 6: Weigh (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecanohydro-3aH-cyclopentazo[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxanecyclopentazo-8-yl]-1-(4-fluorophenyl)hex-1-ol (314-1) (98 mg, 0.19 mmol) and dissolve it in anhydrous methanol. 2M hydrochloric acid (3 mL) was added to 10 mL of alcohol at room temperature. After the addition was complete, the mixture was stirred at room temperature. The reaction was detected by TLC (petroleum ether: ethyl acetate = 3:1). Phosphomolybdic acid was observed as a colorimetric reaction. After the reaction was completed, water and ethyl acetate were added for extraction. The organic phase was separated, dried, concentrated, and purified by column chromatography (petroleum ether: ethyl acetate = 0-50%) to obtain 24-[(4-fluorophenyl)(hydroxy)methyl]cholan-5(6)-ene-3β,4β-diol (314) with a purity of 86.49% (38.5 mg, 0.08 mmol, 42.1%) as a white solid. 1 H NMR (399MHz, Chloroform-d) 67.32-7.28 (m, 2H), 7.01 (t, J=8.5Hz, 2H), 5.66 (s, 1H), 4.63 (t, J=6 .3Hz, 1H), 4.12 (s, 1H), 3.54 (s, 1H), 2.06 (d, J=13.4Hz, 1H), 1.98 (d, J=12.4Hz, 1H), 1.81 (d, J=14 .8Hz, 4H), 1.65 (t, J=7.6Hz, 3H), 1.56 (s, 3H), 1.44 (d, J=10.1Hz, 3H), 1.33 (q, J=10.8, 10.2Hz, 4H ), 1.16 (s, 3H), 1.05 (q, J = 10.0Hz, 5H), 0.96 (d, J = 8.9Hz, 1H), 0.87 (s, 3H), 0.65 (d, J = 2.6Hz, 3H). 13 C NMR (100MHz, Chloroform-d) δ128.83, 127.56, 127.49, 127.41, 115.32, 115.11, 74.18, 74.01, 72.47, 56.86, 55.96, 55.95, 50.12, 42 .29, 39.66, 39.63, 36.88, 35.97, 35.76, 35.66, 32.06, 31.78, 28.22, 25.40, 24.21, 22.38, 22.29, 21.05, 20.50, 18.61, 18.58, 11.84. 19F NMR (376MHz, Chloroform-d) δ-115.22, -115.04--115.50(m).LCMS: [M+MeCN+NH4] + =543.55.

[0784] Example 315

[0785] Preparation of compound 315 24-[hydroxy(2-methylphenyl)methyl]cholan-5(6)-ene-3β,4β-diol

[0786]

[0787] Referring to Example 314, magnesium fluorophenyl bromide was replaced with magnesium o-methylphenyl bromide to obtain compound 24-[hydroxy(2-methylphenyl)methyl]cholan-5(6)-ene-3β,4β-diol with a purity of 91.08% (315). 1 H NMR (399MHz, Chloroform-d) δ7.46 (d, J=7.8Hz, 1H), 7.22 (t, J=7.3Hz, 1H), 7.19-7.10 (m, 2H), 5.71-5.64 (m, 1H ), 4.92 (q, J=5.9, 5.2Hz, 1H), 4.13 (d, J=3.6Hz, 1H), 3.54 (dd, J=10.2, 5.8Hz, 1H), 2.33 (s, 3H), 2.10-1.98 (m, 2 H), 1.93-1.78 (m, 4H), 1.65 (d, J=7.4Hz, 4H), 1.58 (d, J=9.9Hz, 4H), 1.43 (t, J=11.7Hz, 5H), 1.20 (s, 1H), 1.17 ( s, 3H), 1.08 (q, J=10.7, 10.1Hz, 4H), 0.90 (dd, J=6.5, 4.3Hz, 3H), 0.85 (d, J=10.3Hz, 1H), 0.66 (d, J=1.6Hz, 3H). 13C NMR(100MHz,Chloroform-d)δ142.73,130.35,130.34,128.81,127.12,127.09,126.28,125.06,125.04,77.30,.77.27,77.21,72.48,70.92,70.69,56.88,56.05,56.00,50.14,42.31,39.66,38.65,38.48,36.88,35.97,35.87,35.69,32.06,31.79,28.22,25.40,24.22,22.72,21.03,20.51,19.11,18.59,11.85,1.01.LCMS:[M+Na] + =503.4。

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound has the structure shown in formula II-1 or II-2: ； in, R 4a It is H or CH3; In the compound shown in Formula II-2, R 22 For R 21 ; R 21 -L 2 -R C ; Each L 2 It is -(CH2)3-; Each R C Independently for -C(R) 21c (R) 21d )-OH; R 21c For H; R 21d The phenyl group is a phenyl group, wherein the phenyl group is independently unsubstituted or substituted by j R groups. d replace; Each j is independently 1, 2, 3 or 4; Each R d Independently F, Cl or C 1-4 Alkoxy; When the carbon atom marked with # is a chiral carbon atom, it is in the R configuration, S configuration, or a mixture of both.

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, It meets one or more of the following conditions: (1) j is 1; and (2)R d Independently F or C 1-4 Alkyl group.

3. The compound of claim 2 or a pharmaceutically acceptable salt thereof, characterized in that, R 21c For H, R 21d It is a phenyl or fluorophenyl.

4. The compound of claim 2 or a pharmaceutically acceptable salt thereof, characterized in that, -C(R 21c (R) 21d )-OH is , , , , , , or .

5. The compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, R C -C(R) 21c (R) 21d )-OH is , , , , , , , , , or .

6. The compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, R C -C(R) 21c (R) 21d )-OH is , , , , , , , or .

7. The compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-2, characterized in that, -L 2 -R C for , , , , , , , , , or .

8. The compound of claim 7 or a pharmaceutically acceptable salt thereof, characterized in that, -L 2 -R C for , , , , , , or .

9. The compound according to any one of claims 1-2 or a pharmaceutically acceptable salt thereof, characterized in that, R 21 for , or .

10. Any of the compounds described below or a pharmaceutically acceptable salt thereof, characterized in that, , , , or .

11. A pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-10, and at least one pharmaceutical excipient.

12. The use of a compound of any one of claims 1-10 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 11 in the preparation of a medicament for inhibiting the SREBP pathway.

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