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 XXI 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 diseases.

Description

TECHNICAL FIELD

[0001] The present application relates to a steroid compound, its preparation method and application. BACKGROUND

[0002] With the change of lifestyle, including high-calorie food and high-sugar beverage intake, lack of exercise and physical activity, etc., in the global range, metabolic diseases represented by hyperlipidemia, obesity, type 2 diabetes and fatty liver have become an increasingly serious health problem. Among them, fatty liver has become an important cause of chronic liver disease in rich areas of Europe and China. The prevalence of simple liver lipid accumulation in ordinary adults is 10% to 30%, of which 10% to 20% are fatty hepatitis, and the incidence of liver cirrhosis and liver cancer within 10 years is as high as 25%. However, so far, the pathophysiological mechanism of fatty liver has not been fully elucidated, and there is still a lack of effective and specific therapeutic drugs in clinic. It is known that the accumulation of cholesterol, triglycerides and other lipids in blood and liver is the main cause of hyperlipidemia, and hyperlipidemia is an important pathogenic factor for atherosclerosis, stroke and fatty liver disease. Therefore, in order to reduce lipids, the development of new drugs targeting lipid metabolism regulation pathway is becoming an important direction of new metabolic disease drug research and development.

[0003] It is known that the lipid synthesis pathway of mammalian cells is an important factor in regulating the balance of lipid metabolism. The key factor regulating cholesterol and fatty acid synthesis is a class of transcription factor proteins, sterol regulatory element binding proteins (SREBPs). The precursors of this class of proteins are first synthesized in the endoplasmic reticulum (ER), and the precursors are transported to the Golgi by SREBP cleavage-activating protein (SCAP), and then cleaved by two proteases (Site-1 protease (S1P) and Site-2 protease (S2P)) to release the active domain of its N terminus into the nucleus to play a transcription factor role. The SREBP protein cleavage and maturation are strictly regulated by the intracellular sterol (such as cholesterol, 25-hydroxycholesterol) level. When the cell accumulates sufficient cholesterol in the endoplasmic reticulum, cholesterol and SCAP bind and change the conformation of SCAP, causing the SCAP-SREBP complex to bind to the protein Insig (Insulin-induced gene), thereby blocking the transport of SREBP to the Golgi and the subsequent activation of SREBP. On the contrary, the increase of active form of SREBP in the nucleus promotes the synthesis of cell lipids. In addition to cholesterol, 25-hydroxycholesterol (25-HC) is another potent endogenous inhibitor of the SREBP pathway. Unlike cholesterol binding to SCAP, 25-HC directly binds to Insig and induces SCAP and Insig binding.

[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, diabetes, and cardiovascular and cerebrovascular diseases, skin damage, liver cancer, and other diseases.

[0005] For hyperlipidemia, its pathogenesis is mainly caused by factors such as increased lipid synthesis or abnormal lipid transport caused by diet or gene mutation, leading to excessive accumulation of blood cholesterol and fatty acids and other lipids. Currently, statins and fibrates are the main lipid-lowering drugs in clinical practice, and the mechanism of action of statins is to inhibit the cellular cholesterol synthesis pathway while promoting the reverse transport of blood cholesterol. This indicates that targeting key factors of the cellular lipid synthesis pathway is an important means to effectively reduce lipid levels.

[0006] So far, no approved therapeutic drug for fatty liver disease, so it is very important to identify therapeutic targets and develop new effective therapies. The pathogenesis of fatty liver disease is known to involve multiple risk factors, such as steatosis that can be triggered by accumulation of triglycerides in the form of lipid droplets, abnormal increase of cholesterol and fatty acids in cells, which can cause endoplasmic reticulum stress and mitochondrial dysfunction, leading to cell death, inflammation and fibrosis. Among them, free cholesterol accumulation is reported as a key driver for the transition from simple steatosis to aggressive steatohepatitis. Second, the establishment of a fatty liver mouse model, a simple cholesterol-free high-fat diet can only induce steatosis even after a long feeding period, and the addition of 1-2% cholesterol in the diet is a necessary condition to achieve inflammation and fibrosis. Therefore, reducing cholesterol can be a new treatment strategy for fatty liver disease. Studies have shown that SREBP is abnormally activated in fatty liver patients and fatty liver mouse models; deletion or knockout of Scap in mouse liver can eliminate the activation of all SREBPs, thereby preventing the occurrence of fatty liver and hyperlipidemia. In addition, recent studies have shown that endoplasmic reticulum stress-induced SREBP abnormal activation promotes lipogenesis and fatty liver. Therefore, these evidences suggest that reducing liver triglyceride and cholesterol levels by inhibiting the SREBP pathway is an effective strategy to prevent and / or treat metabolic disorders including fatty liver. SUMMARY

[0007] The technical problem to be solved by the present application is to provide a new compound having inhibitory activity on the SREBP pathway.

[0008] The present application also provides a compound represented by formula XXI or a pharmaceutically acceptable salt thereof:

[0009]

[0010] wherein R 3d is H or R 3c ;

[0011] R 3c is -S(O)2OH, -C(O)-CH2CH2COOH, -C(O)-CH2COOH, -C(O)-CH2OH, -C(O)-COOH, -CH2CH2OH or -CH2COOH;

[0012] R 4a is H, F, CH3, CF3 or OH;

[0013] R 4b is H, F, NH2, OH or CF3;

[0014] R 4a and R 4btogether with the carbon atom to which they are attached form

[0015] R 7a is H, F, NH2, CH3, or CF3;

[0016] R 7b is F or OH;

[0017] or R 7a and R 7b together with the carbon atom to which they are attached form

[0018] R 8a is H or F;

[0019] R 14a is H;

[0020] R 22 is or R 21 ;

[0021] R 21 is -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 ;

[0022] each L 1 and L 2 is independently a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one -CH2- moiety of said -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is optionally replaced with -X-;

[0023] L 3 is -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one -CH2- moiety of said -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is replaced with -Y-;

[0024] each X is independently -CHR 5a- or -Y-; 5a R 5b - or -Y-;

[0025] each R 5a and R 5b are each independently F, C 1-4 alkyl or fluoroC 1-4 alkyl;

[0026] each Y is independently -O-, -NH-, -N(C 1-4 alkyl)-, -CH=CH-, or -CHR g -;

[0027] R g is -C 1-4 alkylene-OH;

[0028] each R A is independently -NR 21a R 21b ;

[0029] R B is -N(R 21a )-C(O)R 21b or -N(R 21a )-S(O)2R 21b ;

[0030] R D is -OH, -CH2OH, R C , -CH(CH3)-OH or -C(CH3)2-OH;

[0031] each R C is independently -C(R 21c )(R 21d )-OH, CN, -C(O)R E , NH2, C 1-4 alkoxy, 3-6 membered heterocycloalkyl or 5-10 membered heteroaryl; said 5-10 membered heteroaryl is unsubstituted or substituted by p R c ;

[0032] R E is C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl; wherein said C 1-4 alkyl, C 1-4 alkoxy, C 3-6cycloalkyl and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a substituted;

[0033] each R 21a is independently H, 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 are independently unsubstituted or substituted with m R a substituted;

[0034] each R 21b is independently H, C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl, or 5-10 membered heteroaryl, wherein the C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl are independently unsubstituted or substituted with q R b substituted;

[0035] or, -NR 21a R 21b , R 21a and R 21b together with the nitrogen atom connecting them form a 3-10 membered heterocycloalkyl, wherein the 3-10 membered heterocycloalkyl is unsubstituted or substituted with p R c substituted;

[0036] each m, p and q are each independently 1, 2, 3, 4, or 5;

[0037] each R a , R b and R c are each independently F, Cl, OH, COOH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl, C 1-4 alkoxy,

[0038] R 21c is H, F, C 1-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl, or C 3-6 cycloalkyl;

[0039] R 21d is F, C 2-4 alkyl, fluoroC 1-4 alkyl, C2-4 alkenyl, C 3-6 cycloalkyl, phenyl, 5-6 membered heteroaryl or -L 4 -R 21e wherein the phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted with j R d substituents;

[0040] or, R 21c and R 21d together with the carbon atom connecting them form a C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl;

[0041] L 4 is C 1-4 alkylene;

[0042] R 21e is OH, CN, C 1-4 alkoxy, phenyl or 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted with j R d substituents;

[0043] each j is independently 1, 2, 3 or 4;

[0044] each r is independently 0, 1, 2, 3 or 4;

[0045] each R d and R f is independently F, Cl, OH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl, C 1-4 alkoxy or fluoroC 1-4 alkoxy;

[0046] the number of heteroatoms in the heterocycloalkyl and heteroaryl groups is independently 1, 2, 3 or 4, each heteroatom is independently N, O or S; in formula XXI, the carbon atom marked with * is in the R configuration, the S configuration or a mixture of the two; the carbon atom marked with # is a chiral carbon atom when it is in the R configuration, the S configuration or a mixture of the two; the carbon atom marked with A is a chiral carbon atom when it is in the R configuration, the S configuration or a mixture of the two; the carbon atom marked with B is a chiral carbon atom when it is in the R configuration, the S configuration or a mixture of the two.

[0047] In certain preferred embodiments of the application, certain groups in the compounds of formula XXI, XVI, XIX, XXIII, XXIV or XXVI, or a pharmaceutically acceptable salt thereof, are defined as follows, and the groups not mentioned are as described in any of the embodiments of the application (simply referred to as "in some embodiments" or "in some preferred embodiments").

[0048] In some embodiments, in the compound of Formula XXI as previously described, R 3d is H or R 3c ; R 3c is -S(O)2OH, -C(O)-CH2COOH, -C(O)-CH2OH, -C(O)-COOH, -CH2CH2OH, or -CH2COOH;

[0049] R 4a is H, F, CH3, CF3, or OH;

[0050] R 4b is H, F, OH, CH2OH, CN, COOH, CF3, or cyclopropyl;

[0051] alternatively, R 4a and R 4b , together with the carbon atom to which they are attached, form

[0052] R 7a is H, F, CH3, or CF3; R 7b is F or OH; alternatively, R 7a and R 7b , together with the carbon atom to which they are attached, form

[0053] R 8a is H or F;

[0054] R 14a is H;

[0055] R 22 is or R 21

[0056] R 21 is -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 ;

[0057] each L 1 and L 2independently a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one -CH2- moiety of said -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is optionally replaced with -X-;

[0058] L 3 -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one -CH2- moiety of said -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is replaced with -Y-;

[0059] each X is independently -CHR 5a -, -CR 5a R 5b - or -Y-;

[0060] each R 5a and R 5b are each independently F, C 1-4 alkyl or fluoroC 1-4 alkyl;

[0061] each Y is independently -O-, -NH-, -N(C 1-4 alkyl)-, -CH=CH-, or -CHR g -;

[0062] R g is -C 1-4 alkylene-OH;

[0063] each R A is independently -NR 21a R 21b ;

[0064] R B is -N(R 21a )-C(O)R 21b or -N(R 21a )-S(O)2R 21b ;

[0065] R D is -OH, -CH2OH, R C , -CH(CH3)-OH or -C(CH3)2-OH;

[0066] each R Cindependently -C(R 21c )(R 21d )-OH, CN, -C(O)R E , NH2, C 1-4 alkoxy, 3-6 membered heterocycloalkyl or 5-10 membered heteroaryl; said 5-10 membered heteroaryl is unsubstituted or substituted with p R c substituents;

[0067] R E is C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl; wherein said C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a substituents;

[0068] each R 21a is independently H, C 1-6 alkyl, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein said C 1-6 alkyl, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a substituents;

[0069] each R 21b is independently H, C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl or 5-10 membered heteroaryl, wherein said C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl are independently unsubstituted or substituted with q R b substituents;

[0070] or, in -NR 21a R 21b , R 21a and R 21b together with the nitrogen atom linking them form a 3-10 membered heterocycloalkyl, wherein said 3-10 membered heterocycloalkyl is unsubstituted or substituted with p R c substituents;

[0071] each m, p and q are each independently 1, 2, 3, 4 or 5;

[0072] each Ra R b and R c are each independently F, CI, OH, COOH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl, C 1-4 alkoxy,

[0073] R 21c is H, F, C 1-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl or C 3-6 cycloalkyl;

[0074] R 21d is F, C 2-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl, C 3-6 cycloalkyl, phenyl, 5-6 membered heteroaryl or -L 4 -R 21e wherein said phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted by j R d ;

[0075] Alternatively, R 21c and R 21d together with the carbon atom connecting them form a C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl;

[0076] L 4 is C 1-4 alkylene;

[0077] R 21e is OH, CN, C 1-4 alkoxy, phenyl or 5-6 membered heteroaryl, wherein said phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted by j R d ;

[0078] each j is independently 1, 2, 3 or 4;

[0079] each r is independently 0, 1, 2, 3 or 4;

[0080] each R d and R f is independently F, CI, OH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl or C 1-4 alkoxy;

[0081] the number of heteroatoms in the heterocycloalkyl and heteroaryl groups is independently 1, 2, 3, or 4, each heteroatom is independently N, O, or S;

[0082] In Formula XXI, the carbon atom marked with * is in the R configuration, the S configuration, or a mixture of the two; the carbon atom marked with # is a chiral carbon atom when it is in the R configuration, the S configuration, or a mixture of the two; the carbon atom marked with A is a chiral carbon atom when it is in the R configuration, the S configuration, or a mixture of the two; and the carbon atom marked with B is a chiral carbon atom when it is in the R configuration, the S configuration, or a mixture of the two.

[0083] In some embodiments, in the compound of Formula XXI as previously described, R 3d is H.

[0084] In some embodiments, in the compound of Formula XXI as previously described, R 4a is H; R 4b is H or OH; or, R 4a and R 4b together with the carbon atom to which they are attached form

[0085] In some embodiments, in the compound of Formula XXI as previously described, R 4a is H; R 4b is H.

[0086] In some embodiments, in the compound of Formula XXI as previously described, R 4a is H; R 4b is OH.

[0087] In some embodiments, in the compound of Formula XXI as previously described, R 4a and R 4b together with the carbon atom to which they are attached form

[0088] In some embodiments, in the compound of Formula XXI as previously described, R 7a is H; R 7b is F.

[0089] In some embodiments, in the compound of Formula XXI as previously described, R 7a is F; R 7b is F.

[0090] In some embodiments, in the compound of Formula XXI as previously described, R 7a is H; R 7b is OH.

[0091] In some embodiments, in the compound of Formula XXI as previously described, R7a and R 7b and the carbon atoms to which they are attached form

[0092] In some embodiments, in the compound of Formula XXI as previously described, R 7a and R 7b and the carbon atoms to which they are attached form

[0093] In some embodiments, in the compound of Formula XXI as previously described, R 7a is H; R 7b is NH2.

[0094] In some embodiments, in the compound of Formula XXI as previously described, R 8a is H.

[0095] In some embodiments, in the compound of Formula XXI as previously described in any one of the preceding embodiments, R 22 is

[0096] In some embodiments, in the compound of Formula XXI as previously described in any one of the preceding embodiments, R 22 is R 21 .

[0097] In some embodiments, in the compound of Formula XXI as previously described in any one of the preceding embodiments, each X is independently -CHR 5a - or -CR 5a R 5b -, for example -CHR 5a -.

[0098] In some embodiments, in the compound of Formula XXI as previously described in any one of the preceding embodiments, L 1 is a single bond.

[0099] In some embodiments, in the compound of Formula XXI as previously described in any one of the preceding embodiments, L 1 is -(CH2)2- or -CH2-CHR 5a -.

[0100] In some embodiments, in the compound of Formula XXI as previously described in any one of the preceding embodiments, each R 5a and R 5b are each independently F, methyl, ethyl, or trifluoromethyl.

[0101] In some embodiments, in the compound of Formula XXI as previously described in any one of the preceding embodiments, L 1 is -(CH2)3-.

[0102] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, L 1 It is -(CH2)4-.

[0103] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, L 1 It is -(CH2)5-.

[0104] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, L 1 It is -(CH2)6-.

[0105] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R 21 -L 1 -C(O)R A .

[0106] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R 21 -L 1 -S(O)2R A .

[0107] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R 21 -L 1 -R B .

[0108] In some embodiments, in the compound of formula XXI as described in any of the preceding embodiments, each R 21a H and C independently 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.

[0109] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, m is 1, 2, or 3.

[0110] In some embodiments, in the compound of formula XXI as described in any of the preceding embodiments, each R a It can be F, OH, CN or methoxy group independently.

[0111] In some embodiments, in the compound of formula XXI as described in any of the preceding embodiments, each R 21a H and C independently 1-4 Alkyl, -C 1-4 alkylene -OH, -C 1-4 Alkylene-OC 1-4 Alkyl, fluorinated C1-4 alkyl or cyclopropyl.

[0112] In some embodiments, the compound according to any of the preceding embodiments of Formula XXI is one wherein each R 21a is independently H, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, cyclopropyl or trifluoromethyl.

[0113] In some embodiments, the compound according to any of the preceding embodiments of Formula XXI is one wherein each R 21b is independently H, C 1-6 alkyl, C 6-10 aryl or 5-10 membered heteroaryl, wherein said C 1-6 alkyl, C 6-10 aryl and 5-10 membered heteroaryl are independently unsubstituted or substituted with q R b substituents.

[0114] In some embodiments, the compound according to any of the preceding embodiments of Formula XXI is one wherein each R 21b is independently H, -C 1-4 alkylene-OH, -C 1-4 alkylene-OC 1-4 alkyl, fluoroC 1-4 alkyl, tetrahydro morpholinyl, phenyl, pyridyl, thienyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyrimidinyl or benzopyrazolyl, wherein said phenyl, pyridyl, thienyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyrimidinyl and benzopyrazolyl are independently unsubstituted or substituted with q R b substituents.

[0115] In some embodiments, the compound according to any of the preceding embodiments of Formula XXI is one wherein each R 21b is independently H, -C 1-4 alkylene-OH, -C 1-4 alkylene-OC 1-4 alkyl, fluoroC 1-4 alkyl, tetrahydro morpholinyl, phenyl, wherein said phenyl, are independently unsubstituted or substituted with q R b substituents.

[0116] In some embodiments, the compound according to any of the preceding embodiments of Formula XXI is one wherein q is 1, 2 or 3.

[0117] In some embodiments, the compound of Formula XXI is according to any of the preceding embodiments, wherein each R b is independently F, Cl, OH, COOH, CN, NO2, methyl, trifluoromethyl, methoxy,

[0118] In some embodiments, the compound of Formula XXI is according to any of the preceding embodiments, wherein each R 21b is independently H, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, phenyl,

[0119]

[0120] In some embodiments, the compound of Formula XXI is according to any of the preceding embodiments, wherein each R 21b is independently

[0121] In some embodiments, the compound of Formula XXI is according to any of the preceding embodiments, wherein -NR 21a R 21b is 21a and R 21b together with the nitrogen atom connecting them form wherein said is unsubstituted or substituted with p R c .

[0122] In some embodiments, the compound of Formula XXI is according to any of the preceding embodiments, wherein p is 1, 2, or 3.

[0123] In some embodiments, the compound of Formula XXI is according to any of the preceding embodiments, wherein each Rcis independently F, OH, CN, C 1-4 alkyl or fluoroC 1-4 alkyl.

[0124] In some embodiments, the compound of Formula XXI is according to any of the preceding embodiments, wherein each R c is independently F, OH, CN, methyl or trifluoromethyl.

[0125] In some embodiments, the compound of Formula XXI is according to any of the preceding embodiments, wherein -NR 21a R 21b is 21a and R 21b together with the nitrogen atom connecting them form

[0126] In some embodiments, the compound according to Formula XXI is according to any of the preceding embodiments, wherein each R A is independently -NH2,

[0127]

[0128] In some preferred embodiments, the compound according to Formula XXI is according to any of the preceding embodiments, wherein each R A is independently

[0129]

[0130] In some preferred embodiments, the compound according to Formula XXI is according to any of the preceding embodiments, wherein each R A is independently

[0131] In some preferred embodiments, the compound according to Formula XXI is according to any of the preceding embodiments, wherein -L 1 -C(O)R A is

[0132] In some embodiments, the compound according to Formula XXI is according to any of the preceding embodiments, wherein R B is

[0133] In some preferred embodiments, the compound according to Formula XXI is according to any of the preceding embodiments, wherein R B is

[0134] In some embodiments, the compound according to Formula XXI is according to any of the preceding embodiments, wherein R 21 is -L 2 -R C .

[0135] In some embodiments, the compound according to Formula XXI is according to any of the preceding embodiments, wherein L 2 is a single bond.

[0136] In some embodiments, the compound according to Formula XXI is according to any of the preceding embodiments, wherein L 2 is -CH2-.

[0137] In some embodiments, the compound according to Formula XXI is according to any of the preceding embodiments, wherein L 2 is -(CH2)2-.

[0138] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, L 2 is -(CH2)3-.

[0139] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, L 2 is -(CH2)4-.

[0140] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, L 2 is -(CH2)5-.

[0141] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, L 2 is -(CH2)6-.

[0142] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, each R C is independently -C(R 21c )(R 21d )-OH.

[0143] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, when R C is -C(R 21c )(R 21d )-OH, R 21c is H, R 21d is phenyl, or R 21c and R 21d together with the carbon atom connecting them form a C 3-6 cycloalkyl (e.g., cyclopropyl); wherein the phenyl is optionally substituted with 1 or 2 R d , each R d is independently F or C 1-4 alkoxy.

[0144] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, -C(R C )(R 21c )-OH in R 21d is

[0145] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, when R C is -C(R 21c )(R 21d )-OH, R 21c is H, R 21d is C 2-4 alkyl, fluoroC 1-4 alkyl, C 3-6cycloalkyl, phenyl, fluorophenyl, 5-6 membered heteroaryl, or -L 4 -R 21e .

[0146] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein R C is -C(R 21c )(R 21d )-OH, R 21c is H, and R 21d is C 2-4 alkyl, fluoroC 1-4 alkyl, C 3-6 cycloalkyl, phenyl, 5-6 membered heteroaryl, or -L 4 -R 21e , wherein the phenyl is optionally substituted with 1 or 2 R d , each R d is independently F, OH, CN, or C 1-4 alkoxy.

[0147] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein L 4 is methylene or ethylene.

[0148] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein R 21e is OH, CN, C 1-4 alkoxy, phenyl, or

[0149] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein -C(R C )(R 21c )-OH in R 21d is

[0150]

[0151] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein R C is -C(R 21c )(R 21d )-OH, R 21c is F, and R 21d is C 2-4 alkyl (e.g., ethyl).

[0152] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein -C(R C )(R 21c )-OH in R 21d is

[0153] In some embodiments, in the compound represented by formula XXI 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.

[0154] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, R C -C(R) 21c (R) 21d )-OH is

[0155] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, when R C -C(R) 21c (R) 21d When )-OH, R 21c 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).

[0156] In some embodiments, in the compound represented by formula XXI 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.

[0157] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C -C(R) 21c (R) 21d )-OH is

[0158] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, when R C -C(R) 21c (R) 21dR 21c and R 21d and the carbon atom attaching them together form a cyclopropyl, cyclobutyl or 3-6 cycloalkyl or 3-6 membered heterocycloalkyl (e.g. ).

[0159] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein -C(R C )(R 21c )-OH is 21d R 21c and R 21d and the carbon atom attaching them together form a cyclopropyl, cyclobutyl or

[0160] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein -C(R C )(R 21c )-OH is 21d .

[0161]

[0162] In some preferred embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein -C(R C )(R 21c )-OH is 21d .

[0163]

[0164] In some preferred embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein -C(R C )(R 21c )-OH is 21d .

[0165] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein R C is CN.

[0166] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein R C is -C(O)R E .

[0167] In some embodiments, the compound according to Formula XXI as described in any of the preceding embodiments, wherein R E is C 1-4 alkyl or C3-6 Cycloalkyl.

[0168] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R E It can be methyl, ethyl, or cyclohexyl.

[0169] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C It is NH2.

[0170] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C C 1-4 Alkyl groups, such as methoxy groups.

[0171] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C for For example

[0172] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C for For example

[0173] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C for For example

[0174] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C It is a 5-10 membered heteroaryl group, wherein the 5-10 membered heteroaryl group is unsubstituted or p-shaped. c replace.

[0175] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C The 5-10 member heteroaryl in the definition is

[0176] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C for

[0177] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C It is a 3-6 membered heterocyclic alkyl group.

[0178] In some embodiments, in the compound represented by formula XXI as described in any of the preceding embodiments, R C3-6 membered heterocycloalkyl in the definition is

[0179] In some preferred embodiments, the compound according to Formula XXI as set forth in any one of the preceding embodiments, -L 2 -R C is

[0180]

[0181] In some preferred embodiments, the compound according to Formula XXI as set forth in any one of the preceding embodiments, -L 2 -R C is

[0182] In some embodiments, the compound according to Formula XXI as set forth in any one of the preceding embodiments, R 21 is -L 3 -R D .

[0183] In some embodiments, the compound according to Formula XXI as set forth in any one of the preceding embodiments, L 3 is b is attached to R D .

[0184] In some embodiments, the compound according to Formula XXI as set forth in any one of the preceding embodiments, R D is -OH, -CH2OH, -CH(CH3)-OH or -C(CH3)2-OH.

[0185] In some embodiments, the compound according to Formula XXI as set forth in any one of the preceding embodiments, R 21 -L 3 -R D is

[0186] In some preferred embodiments, the compound according to Formula XXI as set forth in any one of the preceding embodiments, R 21 -L 3 -R D is

[0187]

[0188] In some preferred embodiments, the compound according to Formula XXI as set forth in any one of the preceding embodiments, R 21 -L3 -R D is

[0189] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the *-labeled carbon atom is in the R configuration. 21 is

[0190] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the *-labeled carbon atom is in the R configuration. 22 is

[0191] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the *-labeled carbon atom is in the S configuration.

[0192] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the *-labeled carbon atom is in the S configuration.

[0193] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the *-labeled carbon atom is in a mixture of the S and R configurations, e.g., S configuration:R configuration = 1 : 1.

[0194] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the #-labeled carbon atom is in the R configuration.

[0195] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the #-labeled carbon atom is in the S configuration.

[0196] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the #-labeled carbon atom is in a mixture of the S and R configurations, e.g., S configuration:R configuration = 1 : 1.

[0197] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the A-labeled carbon atom is in the R configuration.

[0198] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the A-labeled carbon atom is in the S configuration.

[0199] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the A-labeled carbon atom is in a mixture of the S and R configurations, e.g., S configuration:R configuration = 1 : 1.

[0200] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the B-labeled carbon atom is in the R configuration.

[0201] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the carbon atom marked by B is in the S configuration.

[0202] In some embodiments, in the compound of Formula XXI as described in any of the preceding embodiments, the carbon atom marked by B is in a mixture of S and R configurations, for example, S configuration:R configuration = 1:1.

[0203] In some embodiments, the compound of Formula XXI is a compound of Formula XXI-1:

[0204] wherein the definitions of #, B, R 4a , R 4b , R 7a , R 7b and R 22 are as described in any embodiment of the present application.

[0205] In some embodiments, the compound of Formula XXI is a compound of Formula XXI-2, XXI-3, or XXI-4:

[0206]

[0207] wherein the definitions of B, R 7a , R 7b and R 22 are as described in any embodiment of the present application.

[0208] In some embodiments, the compound of Formula XXI is any one of the following:

[0209]

[0210]

[0211]

[0212] The present application provides a compound of Formula XVI, or a pharmaceutically acceptable salt thereof:

[0213]

[0214] wherein R 3d is H or R 3c ;

[0215] R 3c is -S(O)2OH, -C(O)-CH2CH2COOH, -C(O)-CH2COOH, -C(O)-CH2OH, -C(O)-COOH, -CH2CH2OH, or -CH2COOH;

[0216] R4a H, F, CH3, CF3, or OH;

[0217] R 4b H, F, NH2, OH, or CF3;

[0218] R 4a and R 4b together with the carbon atom to which they are attached form

[0219] R 5 H or R 5c ; R 5c OH or F;

[0220] R 22 R 21 ; R 21 are as described above for Formula XXI;

[0221] In Formula XVI, the carbon atom marked with an * is in the R configuration, the S configuration, or a mixture of the two; when the carbon atom marked with a # is a chiral carbon atom, it is in the R configuration, the S configuration, or a mixture of the two; when the carbon atom marked with a & is a chiral carbon atom, it is in the R configuration, the S configuration, or a mixture of the two.

[0222] In some embodiments, the compound of Formula XVI as previously described, wherein R 3d is H or R 3c ;

[0223] R 3c -S(O)2OH, -C(O)-CH2COOH, -C(O)-CH2OH, -C(O)-COOH, -CH2CH2OH, or -CH2COOH;

[0224] R 4a H, F, CH3, CF3, or OH;

[0225] R 4b H, F, OH, or CF3;

[0226] R 4a and R 4b together with the carbon atom to which they are attached form

[0227] R 5 H or R 5c ; R 5c OH or F;

[0228] R 22 R 21 ;​​

[0229] R 21 is -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 ;

[0230] each L 1 and L 2 is independently a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one -CH2- moiety of said -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is optionally replaced with -X-;

[0231] L 3 is -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one -CH2- moiety of said -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is replaced with -Y-;

[0232] each X is independently -CHR 5a -, -CR 5a R 5b -, or -Y-;

[0233] each R 5a and R 5b is each independently F, C 1-4 alkyl, or fluoroC 1-4 alkyl;

[0234] each Y is independently -O-, -NH-, -N(C 1-4 alkyl)-, -CH=CH-, or -CHR g -;

[0235] R g is -C 1-4 alkylene-OH;

[0236] each R Aindependently -NR 21a R 21b ;

[0237] R B is -N(R 21a )-C(O)R 21b or -N(R 21a )-S(O)2R 21b ;

[0238] R D is -OH, -CH2OH, R C , -CH(CH3)-OH, or -C(CH3)2-OH;

[0239] each R C is independently -C(R 21c )(R 21d )-OH, CN, -C(O)R E , NH2, C 1-4 alkoxy, 3-6 membered heterocycloalkyl or 5-10 membered heteroaryl; said 5-10 membered heteroaryl is unsubstituted or substituted with p R c ;

[0240] R E is C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, or 3-6 membered heterocycloalkyl; wherein said C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a ;

[0241] each R 21a is independently H, C 1-6 alkyl, C 3-6 cycloalkyl, or 3-6 membered heterocycloalkyl, wherein said C 1-6 alkyl, C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a ;

[0242] each R 21b is independently H, C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl, or 5-10 membered heteroaryl, wherein said C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10Aryl and 5-10 membered heteroaryl are independently unsubstituted or substituted with q R b substituted;

[0243] or, -NR 21a R 21b R 21a and R 21b and the nitrogen atom connecting them together form a 3-10 membered heterocycloalkyl, wherein said 3-10 membered heterocycloalkyl is unsubstituted or substituted with p R c substituted;

[0244] each m, p and q are each independently 1, 2, 3, 4 or 5;

[0245] each R a , R b and R c are each independently F, Cl, OH, COOH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl, C 1-4 alkoxy,

[0246] R 21c is H, F, C 1-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl or C 3-6 cycloalkyl;

[0247] R 21d is F, C 2-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl, C 3-6 cycloalkyl, phenyl, 5-6 membered heteroaryl or -L 4 -R 21e , wherein said phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted with j R d substituted;

[0248] or, R 21c and R 21d and the carbon atom connecting them together form a C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl;

[0249] L 4 is C 1-4 alkylene;

[0250] R 21e is OH, CN, C 1-4 alkoxy, phenyl or 5-6 membered heteroaryl, wherein said phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted with j R dsubstituted;

[0251] each j is independently 1, 2, 3, or 4;

[0252] each r is independently 0, 1, 2, 3, or 4;

[0253] each R d and R f are independently F, Cl, OH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl, or C 1-4 alkoxy;

[0254] the number of heteroatoms in the heterocycloalkyl and heteroaryl groups is independently 1, 2, 3, or 4, each heteroatom being independently N, O, or S;

[0255] in Formula XVI, the carbon atom marked with * is in the R configuration, the S configuration, or a mixture of the two; when the carbon atom marked with # is a chiral carbon atom, it is in the R configuration, the S configuration, or a mixture of the two; when the carbon atom marked with & is a chiral carbon atom, it is in the R configuration, the S configuration, or a mixture of the two.

[0256] In some embodiments, in the compound of Formula XVI as previously described, R 3d is H, -C(O)-CH2OH, -C(O)-CH2COOH, or -C(O)-CH2CH2COOH.

[0257] In some embodiments, in the compound of Formula XVI as previously described, R 3d is H.

[0258] In some embodiments, in the compound of Formula XVI as previously described, R 3d is R 3c .

[0259] In some embodiments, in the compound of Formula XVI as previously described, R 3c is -S(O)2OH.

[0260] In some embodiments, in the compound of Formula XVI as previously described, R 3c is -C(O)-CH2COOH.

[0261] In some embodiments, in the compound of Formula XVI as previously described, R 3c is -C(O)-CH2OH.

[0262] In some embodiments, in the compound of Formula XVI as previously described, R 3c is -C(O)-COOH.

[0263] In some embodiments, in the compound of Formula XVI as previously described, R 3c is -CH2CH2OH.

[0264] In some embodiments, in the compound of Formula XVI as previously described, R 3c is -CH2COOH.

[0265] In some embodiments, in the compound of Formula XVI as previously described, R 4a is H, R 4b is OH.

[0266] In some embodiments, in the compound of Formula XVI as previously described, R 4a is CH3, R 4b is F.

[0267] In some embodiments, in the compound of Formula XVI as previously described, R 4a is F, R 4b is F.

[0268] In some embodiments, in the compound of Formula XVI as previously described, R 4a is H, R 4b is CF3.

[0269] In some embodiments, in the compound of Formula XVI as previously described, R 4a is CH3, R 4b is OH.

[0270] In some embodiments, in the compound of Formula XVI as previously described, R 4a is OH, R 4b is CF3.

[0271] In some embodiments, in the compound of Formula XVI as previously described, R 4a and R 4b together with the carbon atom to which they are attached form

[0272] In some embodiments, in the compound of Formula XVI as previously described, R 4a is H, R 4b is F.

[0273] In some embodiments, in the compound of Formula XVI as previously described, R 4a is CF3, R 4b is CF3.

[0274] In some embodiments, in the compound of Formula XVI as previously described, R 4aand R 4b And the carbon atoms connected to them together form

[0275] In some embodiments, in the compound represented by formula XVI as described above, R 4a For H, R 4b For H.

[0276] In some embodiments, in the compound represented by formula XVI as described above, R 4a For H, R 4b It is NH2.

[0277] In some embodiments, in the compound represented by formula XVI as described above, R 5 For H.

[0278] In some embodiments, in the compound represented by formula XVI as described above, R 5 It is OH.

[0279] In some embodiments, in the compound represented by formula XVI as described above, R 5 It is F.

[0280] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, R 22 for

[0281] In some embodiments, in the compound of formula XVI 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 XXI.

[0282] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, R 21 -L 1 -C(O)R A -L 2 -R C or

[0283] In some embodiments, in the compound represented by formula XVI as described in any of the preceding embodiments, L 1 -(CH2)2- or -CH2-CHR 5a -; better, L 1 It is -(CH2)2-.

[0284] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, each R 21a Independently for H and C 1-4 Alkyl, -C1-4 alkylene-OH, -C 1-4 alkylene-OC 1-4 alkyl, fluoroC 1-4 alkyl or cyclopropyl; preferably, each R 21a independently C 1-4 alkyl.

[0285] In some embodiments, the compound according to Formula XVI is as in any preceding embodiment, wherein each R 21a independently H, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, cyclopropyl or trifluoromethyl; for example, methyl.

[0286] In some embodiments, the compound according to Formula XVI is as in any preceding embodiment, wherein each R 21b independently H, C 1-6 alkyl, C 6-10 aryl or 5-10 membered heteroaryl, wherein the C 1-6 alkyl, C 6-10 aryl and 5-10 membered heteroaryl are independently unsubstituted or substituted with q Rb; preferably, each R 21b independently C 6-10 aryl, wherein the C 6-10 aryl is unsubstituted or substituted with q Rb.

[0287] In some embodiments, the compound according to Formula XVI is as in any preceding embodiment, wherein each R 21b independently H, -C 1-4 alkylene-OH, -C 1-4 alkylene-OC 1-4 alkyl, fluoroC 1-4 alkyl, tetrahydro morpholinyl, phenyl, pyridyl, thienyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyrimidinyl or benzopyrazolyl, wherein the phenyl, pyridyl, thienyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyrimidinyl and benzopyrazolyl are independently unsubstituted or substituted with q R b substituted; preferably, each R 21b independently phenyl, wherein the phenyl is unsubstituted or substituted with q R b substituted.

[0288] In some embodiments, the compound according to Formula XVI is as in any preceding embodiment, wherein q is 1, 2 or 3; for example, 1.

[0289] In some embodiments, the compound of Formula XVI is according to any of the preceding embodiments, wherein each R b is independently F, CI, OH, COOH, CN, N02, methyl, trifluoromethyl, methoxy, for example F.

[0290] In some embodiments, the compound of Formula XVI is according to any of the preceding embodiments, wherein each R 21b is independently

[0291] In some preferred embodiments, the compound of Formula XVI is according to any of the preceding embodiments, wherein each R A is independently for example

[0292] In some preferred embodiments, the compound of Formula XXI is according to any of the preceding embodiments, wherein -L 1 -C(O)R A is for example

[0293] In some embodiments, the compound of Formula XVI is according to any of the preceding embodiments, wherein L 2 is -(CH2)2- or -(CH2)3.

[0294] In some embodiments, the compound of Formula XVI is according to any of the preceding embodiments, wherein each R C is independently -C(R 21c )(R 21d )-OH or

[0295] In some embodiments, the compound of Formula XVI is according to any of the preceding embodiments, wherein when R C is -C(R 21c )(R 21d )-OH, R 21c is H, R 21d is C 2-4 alkyl, fluoroC 1-4 alkyl, C 3-6 cycloalkyl, phenyl, 5-6 membered heteroaryl or -L 4 -R 21e , wherein phenyl is optionally substituted with 1 or 2 R d , each R d is independently F, OH, CN or C 1-4 alkoxy.

[0296] In some embodiments, in the compound of Formula XVI as described in any of the preceding embodiments, when R C is -C(R 21c )(R 21d )-OH, R 21c is H, R 21d is C 2-4 alkyl, fluoroC 1-4 alkyl, C 3-6 cycloalkyl, or phenyl, wherein the phenyl is optionally substituted with 1 or 2 R d , each R d is independently F, Cl, OH, fluoroC 1-4 alkyl, C 1-4 alkoxy, or fluoroC 1-4 alkoxy.

[0297] In some embodiments, in the compound of Formula XVI as described in any of the preceding embodiments, when R C is -C(R 21c )(R 21d )-OH, R 21c and R 21d , together with the carbon atom that connects them, form C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl (e.g. ).

[0298] In some embodiments, in the compound of Formula XVI as described in any of the preceding embodiments, when R C is -C(R 21c )(R 21d )-OH, R 21c and R 21d , together with the carbon atom that connects them, form cyclopropyl, cyclobutyl, or preferably cyclopropyl.

[0299] In some embodiments, in the compound of Formula XVI as described in any of the preceding embodiments, when R C is -C(R 21c )(R 21d )-OH, R 21c is CH3or trifluoromethyl, R 21d is C 2-4 alkyl (e.g., ethyl) or trifluoromethyl.

[0300] In some embodiments, in the compound of Formula XVI as described in any of the preceding embodiments, the -C(R C )(R 21c )-OH in R 21d is

[0301] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, -C(R C )-OH is -C(R 21c )-OH is -C(R 21d )-OH is -C(R 21c )-OH is -C(R 21d )-OH is -C(R d )-OH is -C(R

[0302] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, -C(R C )-OH is -C(R 21c )-OH is -C(R 21d )-OH is -C(R

[0303] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, r is 0.

[0304] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, -C(R 21 )-OH is -C(R

[0305] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, the carbon atom marked with an * is in the R configuration.

[0306] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, the carbon atom marked with an * is in the S configuration.

[0307] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, the carbon atom marked with an * is a mixture of the S and R configurations, e.g. S configuration : R configuration = 1 : 1.

[0308] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, the carbon atom marked with a # in formula XVI is in the R configuration when it is a chiral carbon atom.

[0309] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, the carbon atom marked with a # in formula XVI is in the S configuration when it is a chiral carbon atom.

[0310] In some embodiments, in the compound of formula XVI as described in any of the preceding embodiments, the carbon atom marked with a # in formula XVI is a mixture of the S and R configurations, e.g. S configuration : R configuration = 1 : 1.

[0311] In some embodiments, in the compound of Formula XVI as described in any of the preceding embodiments, the carbon atom marked with & is in the R configuration when it is a chiral carbon atom.

[0312] In some embodiments, in the compound of Formula XVI as described in any of the preceding embodiments, the carbon atom marked with & is in the S configuration when it is a chiral carbon atom.

[0313] In some embodiments, in the compound of Formula XVI as described in any of the preceding embodiments, the carbon atom marked with & is a mixture of S and R configurations, e.g., S configuration:R configuration = 1:1.

[0314] In some embodiments, the compound of Formula XVI is a compound of Formula XVI-1:

[0315]

[0316] wherein the definitions of #, &, 3d , R 4a , R 4b , R 5 and R 22 are as described in any of the embodiments of the application.

[0317] In some embodiments, the compound of Formula XVI is any one of the following compounds:

[0318]

[0319]

[0320]

[0321]

[0322]

[0323] In some embodiments, the compound of Formula XVI is a compound of Formula VII:

[0324]

[0325] wherein R 3d is H or R 3c ; and the definitions of R 3c are as described in any of the embodiments of Formula XXI;

[0326] R 22 is or R 21 ; and the definitions of R 21 are as described in any of the embodiments of Formula XXI;

[0327] In some embodiments, the compound of Formula VII is as previously described, wherein the carbon atom marked with an * is in the R configuration, the S configuration, or a mixture of both.

[0328] In some embodiments, the compound of Formula VII as previously described, wherein R 3d is H.

[0329] In some embodiments, the compound of Formula VII as previously described, wherein R 3d is R 3c ; and R 3c is as defined in any of the embodiments of Formula XXI.

[0330] In some embodiments, the compound of Formula VII as previously described in any of the embodiments, wherein R 22 is

[0331] In some embodiments, the compound of Formula VII as previously described in any of the embodiments, wherein R 22 is R 21 ; and R 21 is as defined in any of the embodiments of Formula XXI.

[0332] In some embodiments, the compound of Formula VII as previously described in any of the embodiments, wherein the carbon atom marked with an * is in the R configuration.

[0333] In some embodiments, the compound of Formula VII as previously described in any of the embodiments, wherein the carbon atom marked with an * is in the S configuration.

[0334] In some embodiments, the compound of Formula VII as previously described in any of the embodiments, wherein the carbon atom marked with an * is in a mixture of the S configuration and the R configuration, e.g., S configuration:R configuration = 1 : 1.

[0335] In some embodiments, the compound of Formula VII is a compound of:

[0336]

[0337] The present application also provides a compound of Formula XIX, or a pharmaceutically acceptable salt thereof:

[0338]

[0339] wherein R 3d is H or R 3c ; and R 3c is as defined in any of the embodiments of Formula XXI.

[0340] R 4a is as defined in any of the embodiments of Formula XXI.

[0341] R 4b is as defined in any of the embodiments of Formula XXI;

[0342] R 5 is H or R 5c ; R 5c is OH or F;

[0343] R 6c is H, F, CH3, or CF3;

[0344] R 6d is CN, NH2, F, COOH, C 1-4 alkoxy (e.g., methoxy), or OH;

[0345] Alternatively, R 6c and R 6d together with the carbon atom to which they are attached form

[0346] R 22 is or R 21 ; R 21 is as defined in Formula XXI;

[0347] In Formula XIX, the carbon atom marked with * is in the R configuration, the S configuration, or a mixture of the two; the carbon atom marked with # is a chiral carbon atom when it is in the R configuration, the S configuration, or a mixture of the two; the carbon atom marked with A is a chiral carbon atom when it is in the R configuration, the S configuration, or a mixture of the two; the carbon atom marked with B is a chiral carbon atom when it is in the R configuration, the S configuration, or a mixture of the two.

[0348] In some embodiments, the compound of Formula XIX as previously described, R 3d is H or R 3c ; R 3c is -S(O)2OH, -C(O)-CH2COOH, -C(O)-CH2OH, -C(O)-COOH, -CH2CH2OH, or -CH2COOH;

[0349] R 4a is H, F, CH3, CF3, or OH;

[0350] R 4b is H, F, OH, or CF3;

[0351] Alternatively, R 4a and R 4b together with the carbon atom to which they are attached form

[0352] R 5 is H or R5c ; R 5c is OH or F;

[0353] R 6c is H, F, CH3or CF3;

[0354] R 6d is CN, F, COOH, C 1-4 alkoxy (e.g., methoxy) or OH;

[0355] or R 6c and R 6d together with the carbon atom to which they are attached form

[0356] R 22 is or R 21 ;

[0357] R 21 is -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 ;

[0358] each L 1 and L 2 is independently a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one -CH2- moiety of said -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is optionally replaced with -X-;

[0359] L 3 is -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one -CH2- moiety of said -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is replaced with -Y-;

[0360] each X is independently -CHR 5a -, -CR 5a R 5b - or -Y-;

[0361] each R 5a and R 5b is each independently F, C 1-4 alkyl or fluoroC 1-4 alkyl;

[0362] each Y is independently -O-, -NH-, -N(C 1-4 alkyl)-, -CH=CH-, or -CHR g -;

[0363] R g is -C 1-4 alkylene-OH;

[0364] each R A is independently -NR 21a R 21b ;

[0365] R B is -N(R 21a )-C(O)R 21b or -N(R 21a )-S(O)2R 21b ;

[0366] R D is -OH, -CH2OH, R C , -CH(CH3)-OH or -C(CH3)2-OH;

[0367] each R C is independently -C(R 21c )(R 21d )-OH, CN, -C(O)R E , NH2, C 1-4 alkoxy, 3-6 membered heterocycloalkyl or 5-10 membered heteroaryl; said 5-10 membered heteroaryl is unsubstituted or substituted with p R c ;

[0368] R E is C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl; wherein said C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a ;

[0369] each R 21a independently H, C 1-6 alkyl, C 3-6 cycloalkyl, or 3-6 membered heterocycloalkyl, wherein said C 1-6 alkyl, C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a substituents;

[0370] each R 21b independently H, C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl, or 5-10 membered heteroaryl, wherein said C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl, and 5-10 membered heteroaryl are independently unsubstituted or substituted with q R b substituents;

[0371] or, in -NR 21a R 21b , R 21a and R 21b together with the nitrogen atom connecting them form a 3-10 membered heterocycloalkyl, wherein said 3-10 membered heterocycloalkyl is unsubstituted or substituted with p R c substituents;

[0372] each m, p, and q are each independently 1, 2, 3, 4, or 5;

[0373] each R a , R b , and R c are each independently F, Cl, OH, COOH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl, C 1-4 alkoxy,

[0374] R 21c is H, F, C 1-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl, or C 3-6 cycloalkyl;

[0375] R 21d is F, C 2-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl, C 3-6 cycloalkyl, phenyl, 5-6 membered heteroaryl, or -L 4 -R21e wherein the phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted with j R d substituents;

[0376] or, R 21c and R 21d together with the carbon atom attaching them form a C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl;

[0377] L 4 is C 1-4 alkylene;

[0378] R 21e is OH, CN, C 1-4 alkoxy, phenyl or 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted with j R d substituents;

[0379] each j is independently 1, 2, 3 or 4;

[0380] each r is independently 0, 1, 2, 3 or 4;

[0381] each R d and R f is independently F, Cl, OH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl or C 1-4 alkoxy;

[0382] the number of heteroatoms in the heterocycloalkyl and heteroaryl is independently 1, 2, 3 or 4, each heteroatom is independently N, O or S;

[0383] in Formula XIX, the carbon atom marked with * is in the R configuration, the S configuration or a mixture of the two; the carbon atom marked with # is a chiral carbon atom when it is in the R configuration, the S configuration or a mixture of the two; the carbon atom marked with A is a chiral carbon atom when it is in the R configuration, the S configuration or a mixture of the two; the carbon atom marked with B is a chiral carbon atom when it is in the R configuration, the S configuration or a mixture of the two.

[0384] In some embodiments, in the compound of Formula XIX as previously described, R 3d is H.

[0385] In some embodiments, in the compound of Formula XIX as previously described, R 4a is H and R 4b is OH.

[0386] In some embodiments, in the compound of Formula XIX as previously described, R 4a is H and R4b H.

[0387] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein R 5 OH.

[0388] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein R 5 F.

[0389] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein R 5 H.

[0390] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein R 6c H, R 6d CN.

[0391] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein R 6c H, R 6d COOH.

[0392] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein R 6c H, R 6d OH.

[0393] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein R 6c H, R 6d OCH3.

[0394] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein R 6c and R 6d together with the carbon atom to which they are attached form

[0395] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein R 22

[0396] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein the carbon atom marked with an * is in the R configuration.

[0397] In some embodiments, the compound according to any of the preceding embodiments of Formula XIX is one wherein the carbon atom marked with an * is in the S configuration.

[0398] ​In some embodiments, in the compound of Formula XIX as described in any of the preceding embodiments, the carbon atom marked * is in a mixture of the S and R configurations, for example S configuration : R configuration = 1 : 1.

[0399] In some embodiments, in the compound of Formula XIX as described in any of the preceding embodiments, the carbon atom marked # is in the R configuration.

[0400] In some embodiments, in the compound of Formula XIX as described in any of the preceding embodiments, the carbon atom marked # is in the S configuration.

[0401] In some embodiments, in the compound of Formula XIX as described in any of the preceding embodiments, the carbon atom marked # is in a mixture of the S and R configurations, for example S configuration : R configuration = 1 : 1.

[0402] In some embodiments, in the compound of Formula XIX as described in any of the preceding embodiments, the carbon atom marked A is in the R configuration.

[0403] In some embodiments, in the compound of Formula XIX as described in any of the preceding embodiments, the carbon atom marked A is in the S configuration.

[0404] In some embodiments, in the compound of Formula XIX as described in any of the preceding embodiments, the carbon atom marked A is in a mixture of the S and R configurations, for example S configuration : R configuration = 1 : 1.

[0405] In some embodiments, in the compound of Formula XIX as described in any of the preceding embodiments, the carbon atom marked B is in the R configuration.

[0406] In some embodiments, in the compound of Formula XIX as described in any of the preceding embodiments, the carbon atom marked B is in the S configuration.

[0407] In some embodiments, in the compound of Formula XIX as described in any of the preceding embodiments, the carbon atom marked B is in a mixture of the S and R configurations, for example S configuration : R configuration = 1 : 1.

[0408] In some embodiments, the compound of Formula XIX is a compound of Formula XIX-1:

[0409]

[0410] wherein the definitions of #, A, B, R 4a , R 4b , R 5 , R 6c and R 6d are as described in any embodiment of the present application.

[0411] In some embodiments, the compound of Formula XIX is any one of the following:

[0412]

[0413]

[0414] The present application also provides a compound of Formula XXIII:

[0415]

[0416] wherein R 3d is H or R 3c ; R 3c is as defined in any one of the aspects of Formula XXI;

[0417] R 4a is as defined in any one of the aspects of Formula XXI;

[0418] R 4b is as defined in any one of the aspects of Formula XXI;

[0419] R 19 is CH2OH, CH2CN, CH2COOH, CH2F or CHF2;

[0420] R 22 is or R 2 1; R 21 is as defined in any one of the aspects of Formula XXI;

[0421] In Formula XXIII, the carbon atom marked with * is in R configuration, S configuration or a mixture of both; the carbon atom marked with # is a chiral carbon atom when it is in R configuration, S configuration or a mixture of both.

[0422] In some embodiments, the compound of Formula XXIII as previously described, R 3d is H or R 3c ; R 3c is -S(O)2OH, -C(O)-CH2COOH, -C(O)-CH2OH, -C(O)-COOH, -CH2CH2OH or -CH2COOH;

[0423] R 4a is H, F, CH3, CF3or OH;

[0424] R 4b is H, F, OH or CF3;

[0425] Alternatively, R 4a and R 4band the carbon atoms to which they are attached collectively form

[0426] R 19 is CH2OH, CH2CN, CH2COOH, CH2F or CHF2;

[0427] R 22 is or R 21 ;

[0428] R 21 is -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 ;

[0429] each L 1 and L 2 is independently a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5- or -(CH2)6-, wherein one -CH2- moiety of said -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5- and -(CH2)6- is optionally replaced with -X-;

[0430] L 3 is -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5- or -(CH2)6-, wherein one -CH2- moiety of said -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5- and -(CH2)6- is replaced with -Y-;

[0431] each X is independently -CHR 5a -, -CR 5a R 5b - or -Y-;

[0432] each R 5a and R 5b is each independently F, C 1-4 alkyl or fluoroC 1-4 alkyl;

[0433] each Y is independently -O-, -NH-, -N(C 1-4 alkyl)-, -CH=CH-, or -CHR g -;

[0434] R g is -C 1-4 alkylene-OH;

[0435] each R A is independently -NR 21a R 21b ;

[0436] R B is -N(R 21a )-C(O)R 21b or -N(R 21a )-S(O)2R 21b ;

[0437] R D is -OH, -CH2OH, R C , -CH(CH3)-OH, or -C(CH3)2-OH;

[0438] each R C is independently -C(R 21c )(R 21d )-OH, CN, -C(O)R E , NH2, C 1-4 alkoxy, 3-6 membered heterocycloalkyl, or 5-10 membered heteroaryl; said 5-10 membered heteroaryl is unsubstituted or substituted with p R c ;

[0439] R E is C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, or 3-6 membered heterocycloalkyl; wherein said C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a ;

[0440] each R 21a is independently H, C 1-6 alkyl, C 3-6 cycloalkyl, or 3-6 membered heterocycloalkyl, wherein said C 1-6 alkyl, C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a ;

[0441] each R 21b independently H, C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl or 5-10 membered heteroaryl, wherein said C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl are independently unsubstituted or substituted by q R b substituents;

[0442] or, in case of -NR 21a R 21b , R 21a and R 21b together with the nitrogen atom linking them form a 3-10 membered heterocycloalkyl, wherein said 3-10 membered heterocycloalkyl is unsubstituted or substituted by p R c substituents;

[0443] each m, p and q are each independently 1, 2, 3, 4 or 5;

[0444] each R a , R b and R c are each independently F, CI, OH, COOH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl, C 1-4 alkoxy,

[0445] R 21c is H, F, C 1-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl or C 3-6 cycloalkyl;

[0446] R 21d is F, C 2-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl, C 3-6 cycloalkyl, phenyl, 5-6 membered heteroaryl or -L 4 -R 21e , wherein said phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted by j R d substituents;

[0447] or, R 21c and R 21d together with the carbon atom linking them form a C 3-6cycloalkyl or 3-6 membered heterocycloalkyl;

[0448] L 4 is C 1-4 alkylene;

[0449] R 21e is OH, CN, C 1-4 alkoxy, phenyl or 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted with j R d substituents;

[0450] each j is independently 1, 2, 3 or 4;

[0451] each r is independently 0, 1, 2, 3 or 4;

[0452] each R d and R f is independently F, Cl, OH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl or C 1-4 alkoxy;

[0453] the number of heteroatoms in the heterocycloalkyl and heteroaryl groups is independently 1, 2, 3 or 4, each heteroatom being independently N, O or S;

[0454] In Formula XXIII, the carbon atom marked with * is in the R configuration, the S configuration or a mixture of the two; the carbon atom marked with # is a chiral carbon atom when it is in the R configuration, the S configuration or a mixture of the two.

[0455] In some embodiments, the compound of Formula XXIII as previously described is one wherein R 3d is H.

[0456] In some embodiments, the compound of Formula XXIII as previously described is one wherein R 4a is H; R 4b is H.

[0457] In some embodiments, the compound of Formula XXIII as previously described in any of the preceding embodiments is one wherein R 22 is

[0458] In some embodiments, the compound of Formula XXIII as previously described in any of the preceding embodiments is one wherein the carbon atom marked with * is in the R configuration.

[0459] In some embodiments, the compound of Formula XXIII as previously described in any of the preceding embodiments is one wherein the carbon atom marked with * is in the S configuration.

[0460] In some embodiments, in the compound of Formula XXIII as described in any of the preceding embodiments, the *-marked carbon atom is in the mixture of S and R configurations, for example, S configuration : R configuration = 1 : 1.

[0461] In some embodiments, in the compound of Formula XXIII as described in any of the preceding embodiments, the #-marked carbon atom is in the R configuration.

[0462] In some embodiments, in the compound of Formula XXIII as described in any of the preceding embodiments, the #-marked carbon atom is in the S configuration.

[0463] In some embodiments, in the compound of Formula XXIII as described in any of the preceding embodiments, the #-marked carbon atom is in the mixture of S and R configurations, for example, S configuration : R configuration = 1 : 1.

[0464] In some embodiments, the compound of Formula XXIII is any one of the following:

[0465]

[0466] The present application also provides a compound of Formula XXIV, or a pharmaceutically acceptable salt thereof:

[0467]

[0468] wherein R 3d is H or R 3c ; R 3c is as defined in any of the embodiments of Formula XXI;

[0469] R 7a is as defined in any of the embodiments of Formula XXI;

[0470] R 7b is as defined in any of the embodiments of Formula XXI;

[0471] R 8a is H or F;

[0472] R 22 is or R 21 ; R 21 is as defined in any of the embodiments of Formula XXI;

[0473] In Formula XXIV, the *-marked carbon atom is in the R configuration, the S configuration, or a mixture of the two; the #-marked carbon atom, when it is a chiral carbon atom, is in the R configuration, the S configuration, or a mixture of the two; and the A-marked carbon atom, when it is a chiral carbon atom, is in the R configuration, the S configuration, or a mixture of the two.

[0474] In some embodiments, in the compound of Formula XXIV as previously described, R 3d is H or R 3c ; R 3c is -S(O)2OH, -C(O)-CH2COOH, -C(O)-CH2OH, -C(O)-COOH, -CH2CH2OH, or -CH2COOH;

[0475] R 7a is H, F, CH3, or CF3; R 7b is F or OH; or, R 7a and R 7b together with the carbon atom to which they are attached form

[0476] R 8a is H or F;

[0477] R 22 is or R 21 ;

[0478] R 21 is -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 ;

[0479] each L 1 and L 2 is independently a single bond, -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one -CH2- moiety of said -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is optionally replaced with -X-;

[0480] L 3 is -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, or -(CH2)6-, wherein one -CH2- moiety of said -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, and -(CH2)6- is replaced with -Y-;

[0481] each X is independently -CHR5a -、-CR 5a R 5b -or-Y-;

[0482] Each R 5a and R 5b Each is independently F and C 1-4 Alkyl or fluorinated C 1-4 alkyl;

[0483] Each Y is independently -O-, -NH-, -N(C) 1-4 alkyl)-, -CH=CH-、 or -CHR g -;

[0484] R g -C 1-4 alkylene-OH;

[0485] Each R A Independently for -NR 21a R 21b ;

[0486] R B -N(R) 21a )-C(O)R 21b or -N(R) 21a )-S(O)2R 21b ;

[0487] R D -OH, -CH2OH, R C -CH(CH3)-OH or -C(CH3)2-OH;

[0488] 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;

[0489] 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 3-6cycloalkyl and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a substituted;

[0490] each R 21a is independently H, 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 are independently unsubstituted or substituted with m R a substituted;

[0491] each R 21b is independently H, C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl, or 5-10 membered heteroaryl, wherein the C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl are independently unsubstituted or substituted with q R b substituted;

[0492] or, in -NR 21a R 21b , R 21a and R 21b together with the nitrogen atom connecting them form a 3-10 membered heterocycloalkyl, wherein the 3-10 membered heterocycloalkyl is unsubstituted or substituted with p R c substituted;

[0493] each m, p and q are each independently 1, 2, 3, 4, or 5;

[0494] each R a , R b and R c are each independently F, Cl, OH, COOH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl, C 1-4 alkoxy,

[0495] R 21c is H, F, C 1-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl, or C 3-6 cycloalkyl;

[0496] R 21d is F, C 2-4 alkyl, fluoroC 1-4 alkyl, C2-4 alkenyl, C 3-6 cycloalkyl, phenyl, 5-6 membered heteroaryl, or -L 4 -R 21e wherein the phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted with j R d substituents;

[0497] or, R 21c and R 21d together with the carbon atom connecting them form a C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl;

[0498] L 4 is C 1-4 alkylene;

[0499] R 21e is OH, CN, C 1-4 alkoxy, phenyl, or 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted with j R d substituents;

[0500] each j is independently 1, 2, 3, or 4;

[0501] each r is independently 0, 1, 2, 3, or 4;

[0502] each R d and R f is independently F, Cl, OH, CN, NO2, C 1-4 alkyl, fluoroC 1-4 alkyl, or C 1-4 alkoxy;

[0503] the number of heteroatoms in the heterocycloalkyl and heteroaryl groups is independently 1, 2, 3, or 4, each heteroatom being independently N, O, or S;

[0504] in Formula XXIV, the carbon atom marked with * is in the R configuration, the S configuration, or a mixture of the two; the carbon atom marked with # is a chiral carbon atom when it is in the R configuration, the S configuration, or a mixture of the two; the carbon atom marked with A is a chiral carbon atom when it is in the R configuration, the S configuration, or a mixture of the two.

[0505] In some embodiments, in the compound of Formula XXIV as previously described, R 3d is H.

[0506] In some embodiments, in the compound of Formula XXIV as previously described, R 7a is H; and R 7b is OH.

[0507] In some embodiments, in the compound of formula XXIV as previously described, R 7a and R 7b together with the carbon atom to which they are attached form

[0508] In some embodiments, in the compound of formula XXIV as previously described, R 8a is H.

[0509] In some embodiments, in the compound of formula XXIV as previously described in any one of the preceding embodiments, R 22 is

[0510] In some embodiments, in the compound of formula XXIV as previously described in any one of the preceding embodiments, the *-labeled carbon atom is in the R configuration.

[0511] In some embodiments, in the compound of formula XXIV as previously described in any one of the preceding embodiments, the *-labeled carbon atom is in the S configuration.

[0512] In some embodiments, in the compound of formula XXIV as previously described in any one of the preceding embodiments, the *-labeled carbon atom is a mixture of the S and R configurations, e.g., S configuration:R configuration = 1:1.

[0513] In some embodiments, in the compound of formula XXIV as previously described in any one of the preceding embodiments, the #-labeled carbon atom is in the R configuration.

[0514] In some embodiments, in the compound of formula XXIV as previously described in any one of the preceding embodiments, the #-labeled carbon atom is in the S configuration.

[0515] In some embodiments, in the compound of formula XXIV as previously described in any one of the preceding embodiments, the #-labeled carbon atom is a mixture of the S and R configurations, e.g., S configuration:R configuration = 1:1.

[0516] In some embodiments, in the compound of formula XXIV as previously described in any one of the preceding embodiments, the A-labeled carbon atom is in the R configuration.

[0517] In some embodiments, in the compound of formula XXIV as previously described in any one of the preceding embodiments, the A-labeled carbon atom is in the S configuration.

[0518] In some embodiments, in the compound of formula XXIV as previously described in any one of the preceding embodiments, the A-labeled carbon atom is a mixture of the S and R configurations, e.g., S configuration:R configuration = 1:1.

[0519] In some embodiments, the compound of formula XXIV is any one of the following:

[0520]

[0521] The present application also provides a compound of Formula XXVI:

[0522]

[0523] R 3a and R 3b are defined as described in any one of the following groups:

[0524] (i) R 3a is H, R 3b is -NH2, -NHS(O)2CH3, -NHS(O)2CH2CH3, -NHS(O)2CH2CH2CH3, -NHS(O)2CF3, -NH-C(O)-CH2OH, -CH2OH, -COOH, -C(O)-OCH3, -CH(CH3)OH, -CH2-C(O)-CH2OH, or -CH(OH)-CH2CH2OH;

[0525] (ii) R 3a is CH3, R 3b is -OH; and

[0526] (iii) R 3a and R 3b together with the carbon atom to which they are attached form

[0527] R 4a is defined as described in any one of the embodiments of Formula XXI;

[0528] R 4b is defined as described in any one of the embodiments of Formula XXI;

[0529] R 5 is defined as described in any one of the embodiments of Formula XVI;

[0530] R 22 is or R 21 ; R 21 is defined as described in any one of the embodiments of Formula XXI;

[0531] In Formula XXVI, the carbon atom marked with an * is in the R configuration, the S configuration, or a mixture of the two; when the carbon atom marked with a # is a chiral carbon atom, it is in the R configuration, the S configuration, or a mixture of the two; when the carbon atom marked with a & is a chiral carbon atom, it is in the R configuration, the S configuration, or a mixture of the two.

[0532] In some embodiments, the compound of Formula XXVI as previously described, R 3a and R3b The definition is as described in any of the following groups:

[0533] (i)R 3a For H, R 3b is -NH2, -NHS(O)2CH3, -NHS(O)2CH2CH3, -NHS(O)2CH2CH2CH3, -NHS(O)2CF3, -NH-C(O)-CH2OH, -CH2OH, -COOH, -C(O)-OCH3, -CH(CH3)OH, -CH2-C(O)-CH2OH or -CH(OH)-CH2CH2OH;

[0534] (ii)R 3a CH3, R 3b -OH; and

[0535] (iii)R 3a and R 3b And the carbon atoms connected to them together form

[0536] R 4a It can be H, F, CH3, CF3 or OH;

[0537] R 4b It can be H, F, OH or CF3;

[0538] Or, R 4a and R 4b And the carbon atoms connected to them together form

[0539] R 5 For H or R 5c ;R 5c It is OH or F;

[0540] R 22 for or R 21 ;

[0541] 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 ;

[0542] Each L 1 and L2 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-;

[0543] 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-;

[0544] Each X is independently -CHR 5a -、-CR 5a R 5b -or-Y-;

[0545] Each R 5a and R 5b Each is independently F and C 1-4 Alkyl or fluorinated C 1-4 alkyl;

[0546] Each Y is independently -O-, -NH-, -N(C) 1-4 alkyl)-, -CH=CH-、 or -CHR g -;

[0547] R g -C 1-4 alkylene-OH;

[0548] Each R A Independently for -NR 21a R 21b ;

[0549] R B -N(R) 21a )-C(O)R 21b or -N(R) 21a )-S(O)2R 21b ;

[0550] R D -OH, -CH2OH, R C -CH(CH3)-OH or -C(CH3)2-OH;

[0551] Each RC independently -C(R 21c )(R 21d )-OH, CN, -C(O)R E , NH2, C 1-4 alkoxy, 3-6 membered heterocycloalkyl or 5-10 membered heteroaryl; said 5-10 membered heteroaryl is unsubstituted or substituted with p R c substituents;

[0552] R E is C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl; wherein said C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a substituents;

[0553] each R 21a is independently H, C 1-6 alkyl, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein said C 1-6 alkyl, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are independently unsubstituted or substituted with m R a substituents;

[0554] each R 21b is independently H, C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl or 5-10 membered heteroaryl, wherein said C 1-6 alkyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, C 6-10 aryl and 5-10 membered heteroaryl are independently unsubstituted or substituted with q R b substituents;

[0555] or, in -NR 21a R 21b , R 21a and R 21b together with the nitrogen atom linking them form a 3-10 membered heterocycloalkyl, wherein said 3-10 membered heterocycloalkyl is unsubstituted or substituted with p R c substituents;

[0556] each m, p and q are each independently 1, 2, 3, 4 or 5;

[0557] each R a , R b and R c is independently F, CI, OH, COOH, CN, N02, C 1-4 alkyl, fluoroC 1-4 alkyl, C 1-4 alkoxy,

[0558] R 21c is H, F, C 1-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl or C 3-6 cycloalkyl;

[0559] R 21d is F, C 2-4 alkyl, fluoroC 1-4 alkyl, C 2-4 alkenyl, C 3-6 cycloalkyl, phenyl, 5-6 membered heteroaryl or -L 4 -R 21e wherein said phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted by j R d ;

[0560] or, R 21c and R 21d together with the carbon atom connecting them form a C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl;

[0561] L 4 is C 1-4 alkylene;

[0562] R 21e is OH, CN, C 1-4 alkoxy, phenyl or 5-6 membered heteroaryl, wherein said phenyl and 5-6 membered heteroaryl are independently unsubstituted or substituted by j R d ;

[0563] each j is independently 1, 2, 3 or 4;

[0564] each r is independently 0, 1, 2, 3 or 4;

[0565] each R d and R f is independently F, CI, OH, CN, N02, C 1-4 alkyl, fluoroC 1-4 alkyl or C 1-4 alkoxy;

[0566] the number of heteroatoms in said heterocycloalkyl and heteroaryl groups is independently 1, 2, 3, or 4, each heteroatom is independently N, O, or S;

[0567] In Formula XXVI, the *-labeled carbon atom is in the R configuration, the S configuration, or a mixture of the two; when the #-labeled carbon atom is a chiral carbon atom, it is in the R configuration, the S configuration, or a mixture of the two; when the &-labeled carbon atom is a chiral carbon atom, it is in the R configuration, the S configuration, or a mixture of the two.

[0568] In some embodiments, in the compound of Formula XXVI as previously described, R 3a and R 3b are as described in any one of the following groups:

[0569] a) R 3a is H, and R 3b is -NHS(O)2CH3, -CH2OH, -COOH, -C(O)-OCH3, or -CH(CH3)OH; and

[0570] b) R 3a is CH3, and R 3b is -OH.

[0571] In some embodiments, in the compound of Formula XIX as previously described, R 4a is H, and R 4b is H.

[0572] In some embodiments, in the compound of Formula XXVI as previously described, R 5 is H.

[0573] In some embodiments, in the compound of Formula XXVI as previously described, R 22 is

[0574] In some embodiments, in the compound of Formula XXVI as previously described in any one of the preceding embodiments, the *-labeled carbon atom is in the R configuration.

[0575] In some embodiments, in the compound of Formula XXVI as previously described in any one of the preceding embodiments, the *-labeled carbon atom is in the S configuration.

[0576] In some embodiments, in the compound of Formula XXVI as previously described in any one of the preceding embodiments, the *-labeled carbon atom is in a mixture of the S configuration and the R configuration, for example, S configuration:R configuration = 1:1.

[0577] In some embodiments, in the compound of Formula XXVI as previously described in any one of the preceding embodiments, in Formula XXVI, when the #-labeled carbon atom is a chiral carbon atom, it is in the R configuration.

[0578] In some embodiments, in the compound of Formula XXVI as described in any of the preceding embodiments, the carbon atom marked with # is a chiral carbon atom, and when it is in the S configuration.

[0579] In some embodiments, in the compound of Formula XXVI as described in any of the preceding embodiments, the carbon atom marked with # is a chiral carbon atom, and when it is in the S configuration.

[0580] In some embodiments, in the compound of Formula XXVI as described in any of the preceding embodiments, the carbon atom marked with # is a chiral carbon atom, and when it is in the S configuration.

[0581] In some embodiments, in the compound of Formula XXVI as described in any of the preceding embodiments, the carbon atom marked with # is a chiral carbon atom, and when it is in the S configuration.

[0582] In some embodiments, in the compound of Formula XXVI as described in any of the preceding embodiments, the carbon atom marked with # is a chiral carbon atom, and when it is in the S configuration.

[0583] In some embodiments, the compound of Formula XXVI is a compound of Formula XXVI-1:

[0584]

[0585] wherein the definitions of *, R 3a and R 3b are as described in any embodiment of the present application.

[0586] In some embodiments, the compound of Formula XXVI is any one of the following compounds:

[0587]

[0588] The compounds of the present application can be prepared from known starting materials (e.g., lanosterol) by various conventional reaction methods (e.g., hydroxyl protection, double bond ozonolysis, wittig reaction, hydrolysis reaction, amide condensation reaction, Grignard reagent addition reaction, reduction reaction, nucleophilic substitution reaction, epoxidation reaction) in the art. Exemplary preparation methods are described in the preparation examples of the present application.

[0589] For example, the preparation method of compound 37 and its analogs can be prepared from lanosterol as the starting material, through hydroxyl protection, double bond ozonolysis, wittig reaction, hydrolysis, condensation to obtain compound 37 and its analogs; the reaction scheme is as shown below:

[0590]

[0591] For example, the preparation method of compound 101 and its analogs can take lanosterol as the starting material, and the compound 101 and its analogs can be obtained through hydroxyl protection, double bond ozone oxidation, Wittig reaction, hydrolysis and Grignard reagent addition reaction; the reaction route is shown in the following formula:

[0592]

[0593] For example, the preparation method of compound 80 and its analogs can take lanosterol as the starting material, and the compound 80 and its analogs can be obtained through hydroxyl protection, double bond ozone oxidation, reduction, two-step substitution and addition reaction; the reaction route is shown in the following formula:

[0594]

[0595] For example, the preparation method of compound 125 and its analogs can take lanosterol as the starting material, and the compound 125 and its analogs can be obtained through hydroxyl protection, double bond ozone oxidation, Wittig reaction, hydrolysis and epoxidation; the reaction route is shown in the following formula:

[0596]

[0597] For example, the preparation method of compound 193 and its analogs can take lanosterol as the starting material, and the compound 193 can be obtained through hydroxyl protection, double bond ozone oxidation, double bond shift, double bond ozone oxidation, reduction amination, condensation and hydrolysis; the reaction route of the preparation method is shown in the following formula:

[0598]

[0599] The application also provides a pharmaceutical composition comprising the compound or the pharmaceutically acceptable salt thereof as described above, and at least one pharmaceutical excipient.

[0600] The application also provides use of the compound or the pharmaceutically acceptable salt thereof as described above or the pharmaceutical composition as described above in the preparation of a drug for preventing and / or treating diseases, wherein the diseases are obesity, hyperlipidemia, fatty liver, diabetes, atherosclerosis, cardiovascular and cerebrovascular diseases, liver cancer or skin damage.

[0601] The application also provides use of the compound or the pharmaceutically acceptable salt thereof as described above or the pharmaceutical composition as described above in the preparation of a drug for inhibiting the SREBP pathway.

[0602] The application also provides a method for inhibiting the SREBP pathway, which comprises administering an effective amount of the compound or the pharmaceutically acceptable salt thereof as described above to a subject.

[0603] The present application also provides a method for preventing and / or treating a disease, comprising administering to a subject an effective amount of a compound or a pharmaceutically acceptable salt thereof as described above, wherein the disease is obesity, hyperlipemia, fatty liver, diabetes, atherosclerosis, cardiovascular and cerebrovascular diseases, liver cancer or skin damage.

[0604] Definitions and Descriptions

[0605] The following terms and phrases as used herein are intended to have the following meanings unless otherwise indicated. A particular term or phrase should not be construed as indefinite or unclear if not specifically defined, but should be understood according to its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding product or active ingredient thereof.

[0606] In the present text, the term "substituted" or "substituent" means that a hydrogen atom in a group is replaced by a designated group. When the position of substitution is not specified, the substitution can be at any position, but only a stable or chemically feasible compound is allowed. This is illustrated as follows: The structure means that a hydrogen atom on the benzene ring is replaced by q R 8 substituted, when there are multiple R 8 , each R 8 is the same or different.

[0607] When any variable (for example, R) occurs more than one time in a compound or a structure, each occurrence of that variable is independent. Thus, for example, if a group is substituted with 0-2 R, then the group can optionally be substituted up to two times with R, and each occurrence of R is selected independently. In addition, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0608] When the linking group listed herein is not specified in its connection direction, its connection direction can be either from left to right or from right to left. This is illustrated as follows, -A-L-B, where the linking group L is -C-D-, -A-L-B includes -A-C-D-B and -A-D-C-B when the connection direction of L is not specified.

[0609] When one of the variables is selected from a single bond, it means that the two groups connected by it are directly connected, such as L represents a single bond in A-L-Z, which means that the structure is actually A-Z.

[0610] In the present text, the term "alkyl" refers to a saturated straight-chain or branched-chain monovalent hydrocarbon group. C1-C6alkyl means an alkyl group having 1 to 6 carbon atoms. In some embodiments, C1-C6alkyl can be C1-C4alkyl. C1-C4alkyl includes methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, t-butyl.

[0611] In the present text, the term "alkenyl" refers to a straight-chain or branched-chain 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 attached to the remainder of the molecule through either a saturated or unsaturated carbon atom. C2-C4alkenyl means an alkenyl group having 2, 3, or 4 carbon atoms. Specific examples of alkenyl groups include, but are not limited to, ethenyl, allyl.

[0612] In the present text, the term "alkylene" refers to a saturated straight-chain or branched-chain divalent hydrocarbon group. C1-C4alkylene means an alkylene group having 1 to 4 carbon atoms, which specifically is methylene, ethylene (e.g., -CH2CH2-, -CH(CH3)-), propylene (e.g., -CH2CH2CH2-, -C(CH3)2-, -CH2CH(CH3)-), butylene (e.g., -CH2CH2CH2CH2-, -CH(CH3)CH(CH3)-, -CH2CH(CH3)CH2-).

[0613] In the present text, the term "fluoroalkyl" refers to a group in which one or more hydrogen atoms in an alkyl group, as previously defined, are replaced by fluorine. Examples of fluoroalkyl groups include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl.

[0614] In the present text, the term "alkoxy" refers to -O-alkyl, wherein alkyl is as previously defined. C1-C4alkoxy means -O-(C1-C4alkyl), wherein C1-C4alkyl is as previously defined, i.e., C1-C4alkoxy specifically can be methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, sec-butoxy, or t-butoxy.

[0615] In the present text, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic (e.g., fused, spiro, or bridged) cyclic hydrocarbon group. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, C 3-10 Cycloalkyl specifically can be C3, C4, C5, C6, C7, C8, C9, C 10 Cycloalkyl specifically can be C3, C4, C5, C6, C7, C8, C9, C 3-6 Cycloalkyl specifically can be C3, C4, C5, C6, C7, C8, C9, C

[0616] In the present text, the term "heterocycloalkyl" refers to a saturated monocyclic or polycyclic (e.g. fused, spiro or bridged) ring-like group formed by carbon atoms and at least one heteroatom, wherein the heteroatom is independently selected from N, O and S. The heterocycloalkyl group can be attached to the rest of the structure through a carbon atom and a heteroatom of the ring. Examples of heterocycloalkyl groups include, but are not limited to tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl. A 3-10 membered heterocycloalkyl group can in particular be a 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-membered heterocycloalkyl group. A 3-6 membered heterocycloalkyl group can in particular be a 3-, 4-, 5- or 6-membered heterocycloalkyl group. In some embodiments, the heterocycloalkyl group is monocyclic. In some embodiments, the heterocycloalkyl group is polycyclic (e.g. fused, spiro or bridged).

[0617] In the present text, the term "C 6-10 aryl" refers to phenyl or naphthyl.

[0618] In the present text, the term "heteroaryl" refers to an aromatic monocyclic or fused ring group formed by carbon atoms and at least one heteroatom, wherein the heteroatom is independently selected from N, O and S. A 5-10 membered heteroaryl group can in particular be a 5-, 6-, 7-, 8-, 9- or 10-membered heteroaryl group, e.g. a 5-6 membered heteroaryl group or an 8-10 membered fused heteroaryl group. A 5-6 membered heteroaryl group is monocyclic, particular examples include but are not limited to pyrrole, furan, thiophene, oxazole, isoxazole, thiazole, isothiazole, pyrazole, imidazole, pyridine, pyrimidine, pyrazine. Examples of 8-10 membered fused heteroaryl groups include but are not limited to benzopyrrole, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazole, benzoisothiazole, benzopyrazole, benzoimidazole, benzopyridine, benzopyrimidine, benzopyrazine, thiazolothiazole, pyridopyridine, pyridopyrazine, pyridopyrimidine.

[0619] In the present text, the chemical structure formulae indicate the position of attachment. When included in a cyclic group and no ring atom to which it is attached is indicated, may be attached to any ring atom, but only those that form a stable or chemically feasible compound are allowed. For example, include structures.

[0620] 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.

[0621] In chemical structures, wedge-shaped solid lines 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 Or simultaneously include and Two configurations (e.g.) and 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.

[0622] In this document, the term "subject" includes any animal, preferably a mammal, and more preferably a human.

[0623] The term "effective amount" as used herein refers to a sufficient amount of a drug or pharmaceutical agent to achieve the desired effect without being toxic to the recipient. The determination of an effective amount is dependent on the age and general condition of the recipient, as well as on the particular active ingredient, and an appropriate effective amount for a given case can be determined by a person skilled in the art according to routine experiments.

[0624] The above-mentioned preferred conditions can be combined in any manner without departing from the common general knowledge in the art, thereby obtaining various preferred embodiments of the present application.

[0625] The reagents and raw materials used in the present application are commercially available.

[0626] The positive progress effect of the present application is that the present application provides a new class of compounds which have inhibitory activity on the SREBP pathway, 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. BRIEF DESCRIPTION OF DRAWINGS

[0627] Figure 1 Results of 25-hydroxy lanosterol inhibiting weight gain of mice induced by AMLN diet.

[0628] Figure 2 Effects of 25-hydroxy lanosterol on food intake of mice.

[0629] Figure 3 25-hydroxy lanosterol reduces the content of total cholesterol in the blood of mice.

[0630] Figure 4 25-hydroxy lanosterol reduces the content of total triglyceride in the blood of mice.

[0631] Figure 5 25-hydroxy lanosterol reduces the content of total cholesterol in the liver of mice.

[0632] Figure 6 25-hydroxy lanosterol reduces the content of total triglyceride in the liver of mice.

[0633] Figure 7 25-hydroxy lanosterol reduces the level of aspartate aminotransferase (AST) in the blood of mice.

[0634] Figure 8 25-hydroxy lanosterol reduces the level of alanine aminotransferase (ALT) in the blood of mice.

[0635] Figure 9 HE staining results of liver sections of mice: C57BL / 6J CD group.

[0636] Figure 10 HE staining results of liver sections of mice: Ldlr - / -CD group.

[0637] Figure 11 HE staining results of mouse liver sections: Ldlr - / - AMLN group.

[0638] Figure 12 HE staining results of mouse liver sections: Ldlr - / - AMLN+25-HL group.

[0639] Figure 13 HE staining results and NAFLD activity score quantitative statistics of liver sections of 4 groups of mice.

[0640] Figure 14 Oil red O staining results of mouse liver sections: C57BL / 6J CD group.

[0641] Figure 15 Oil red O staining results of mouse liver sections: Ldlr - / - CD group.

[0642] Figure 16 Oil red O staining results of mouse liver sections: Ldlr - / - AMLN group.

[0643] Figure 17 Oil red O staining results of mouse liver sections: Ldlr - / - AMLN+25-HL group.

[0644] Figure 18 Quantitative results of oil red O staining of liver sections of 4 groups of mice.

[0645] Figure 19 Sirius red staining results of mouse liver sections: C57BL / 6J CD group.

[0646] Figure 20 Sirius red staining results of mouse liver sections: Ldlr - / - CD group.

[0647] Figure 21 Sirius red staining results of mouse liver sections: Ldlr - / - AMLN group.

[0648] Figure 22 Sirius red staining results of mouse liver sections: Ldlr - / - AMLN+25-HL group.

[0649] Figure 23 Quantitative results of Sirius red staining of liver sections of 4 groups of mice.

[0650] Figure 24F4 / 80 immunostaining results for mouse liver sections, and polarized light imaging results indicating cholesterol crystallization: C57BL / 6J CD group.

[0651] Figure 25 F4 / 80 immunostaining results for mouse liver sections, and polarized light imaging results indicating cholesterol crystallization: Ldlr - / - CD group.

[0652] Figure 26 F4 / 80 immunostaining results for mouse liver sections, and polarized light imaging results indicating cholesterol crystallization: Ldlr - / - AMLN group.

[0653] Figure 27 F4 / 80 immunostaining results for mouse liver sections, and polarized light imaging results indicating cholesterol crystallization: Ldlr - / - AMLN + 25-HL group.

[0654] Figure 28 Quantitative results of F4 / 80 immunostaining for liver sections of 4 groups of mice.

[0655] Figure 29 Quantitative results of polarized light imaging indicating cholesterol crystallization for 4 groups of mice.

[0656] Figure 30 Sudan IV staining results for mouse aortic tree: C57BL / 6J CD group.

[0657] Figure 31 Sudan IV staining results for mouse aortic tree: Ldlr - / - CD group.

[0658] Figure 32 Sudan IV staining results for mouse aortic tree: Ldlr - / - AMLN group.

[0659] Figure 33 Sudan IV staining results for mouse aortic tree: Ldlr - / - AMLN + 25-HL group.

[0660] Figure 34 Quantitative results of Sudan IV staining for aortic tree of 4 groups of mice.

[0661] Figure 35 25-Hydroxy lanosterol reduces the expression of genes related to lipid production (Hmgcs, Hmgcr and SCD1, FASN) in the liver organoids.

[0662] Figure 36Decreased expression of liver organoids and fibrosis-related genes (Col1a1, aSMA) for 25-hydroxy lanoestrone.

[0663] Figure 37 Brightfield imaging results for mouse liver organoids: vehicle control group.

[0664] Figure 38 Brightfield imaging results for mouse liver organoids: obeticholic acid 1 mM group.

[0665] Figure 39 Brightfield imaging results for mouse liver organoids: obeticholic acid 3 mM group.

[0666] Figure 40 Brightfield imaging results for mouse liver organoids: 25-HL 1 mM group.

[0667] Figure 41 Brightfield imaging results for mouse liver organoids: 25-HL 3 mM group.

[0668] Figure 42 Nile red staining results for mouse liver organoids: vehicle control group.

[0669] Figure 43 Nile red staining results for mouse liver organoids: obeticholic acid 1 mM group.

[0670] Figure 44 Nile red staining results for mouse liver organoids: obeticholic acid 3 mM group.

[0671] Figure 45 Nile red staining results for mouse liver organoids: 25-HL 1 mM group.

[0672] Figure 46 Nile red staining results for mouse liver organoids: 25-HL 3 mM group.

[0673] Figure 47 Nile red staining quantification results for 5 groups of mouse liver organoids.

[0674] Figure 48 Immunofluorescence staining results for fibrosis marker protein aSMA for mouse liver organoids: vehicle control group.

[0675] Figure 49 Immunofluorescence staining results for fibrosis marker protein aSMA for mouse liver organoids: obeticholic acid 1 mM group.

[0676] Figure 50 Immunofluorescence staining results for fibrosis marker protein aSMA for mouse liver organoids: obeticholic acid 3 mM group.

[0677] Figure 51 Fibrosis marker protein aSMA immunofluorescence staining results of mouse liver organoids: 25-HL 1 μΜ group.

[0678] Figure 52 Fibrosis marker protein aSMA immunofluorescence staining results of mouse liver organoids: 25-HL 3 μΜ group.

[0679] Figure 53 Fibrosis marker protein aSMA immunofluorescence staining quantitative results of 5 groups of mouse liver organoids.

[0680] Figure 54 Full spectrum of 2D NMR spectrum of compound 151-7.

[0681] Figure 55 Local magnification spectrum of 2D NMR spectrum of compound 151-7.

[0682] Figure 56 Full spectrum of 2D NMR spectrum of compound 151,

[0683] Figure 57 Local magnification spectrum of 2D NMR spectrum of compound 151.

[0684] Figure 58 Full spectrum of 2D NMR spectrum of compound 203,

[0685] Figure 59 Local magnification spectrum of 2D NMR spectrum of compound 203.

[0686] Figure 60 2D NMR spectrum of compound 206 Figure 2 Full spectrum,

[0687] Figure 61 Local magnification spectrum of 2D NMR spectrum of compound 206. DETAILED DESCRIPTION

[0688] The present application is further illustrated by the following examples without thereby limiting the present application to the examples described. The experimental methods in the following examples, if no specific conditions are indicated, are selected according to the conventional methods and conditions, or according to the product instructions.

[0689] In the following examples, 25-hydroxy lanosterol (25-HL) refers to the compound of Example 68.

[0690] Biological test examples

[0691] Animals: Adult male C57BL / 6J mice were purchased from Shanghai Slac Laboratory Animal Co., Ltd. Low-density lipoprotein receptor knockout mice (Ldlr - / -Mice were purchased from Jiangsu Jiecui Yekang Biosciences Co., Ltd. (T001464). Mice were maintained under pathogen-free conditions, under 12-hour light / dark conditions, with free access to water and chow. Gavage vehicle was 0.5% Tween-80, 0.5% methylcellulose, 0.9% sodium chloride. AMLN chow (AMLN, Dyets) contained 40% (kcal%) fat (80% of which was trans fat), 22% (mass) fructose, and 2% (mass) cholesterol.

[0692] Reagents: Methylmalonic acid (41288), paraformaldehyde (P6148), Tween-80 (P8074), methylcellulose (V900506), oil red O (O0625), and DAPI for nuclear staining were purchased from Sigma-Aldrich. Lovastatin (purity > 98.5%, HPLC) was purchased from Shanghai Pharmco. Dulbecco’s modified eagle medium (DMEM) for cell culture was purchased from Thermo Scientific, fetal bovine serum (S1580) was purchased from Biowest. Lipoprotein-depleted serum (LPDS) was prepared in our lab by ultracentrifugation. Obeticholic acid (OCA, CAS registry number 459789-99-2, purity 98% (HPLC). Total cholesterol and total triglyceride kits were purchased from Shanghai Kewei Bioengineering Co., Ltd. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) kits were purchased from Lai Er Bio-tech. Hematoxylin-eosin staining kit (6765001, 6766010) was purchased from Thermo Scientific. Sirius red staining kit (ab150681) was purchased from Abeam. Sudan red IV (A610914) was purchased from Sangon Biotech (Shanghai) Co., Ltd. Nile red (HY-D0718) was purchased from MCE.

[0693] Antibodies: The antibodies used for immunofluorescence staining analysis were as follows: anti-alpha smooth muscle Actin (aSMA, ab7817, 1:500) antibody was purchased from Abeam; secondary antibody, fluorescein (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 secondary antibody, Alexa Fluor Plus 488-conjugated goat anti-rat IgG (A-11006, 1:500) were purchased from Invitrogen.

[0694] Biological test example 1: cell culture

[0695] Human hepatocellular carcinoma cell line Huh-7 / SRE-Luc was grown in DMEM containing 10% fetal bovine serum, 100 units / ml penicillin, 100 μg / ml streptomycin and 200 μg / ml G418.

[0696] Biological test example 2: SREBP luciferase reporter system

[0697] Huh-7 / SRE-Luc cell line is a human hepatocellular carcinoma cell line Huh-7 stably expressing LDLR promotor-luciferase and green fluorescent protein (GFP). The LDLR promotor region contains sterol-regulatory-element (SRE) which can be effectively and sensitively regulated by transcription factor SREBP, and the GFP signal is used as an internal control to indicate the change of cell number. Therefore, this cell line can be used to screen active small molecules that regulate SREBP signaling pathway. In our work, we used this cell line to screen inhibitors that can effectively inhibit the SREBP signaling pathway. Cells were incubated in a culture medium lacking sterols (5% delipidated serum, 2 μΜ lovastatin, 10 μΜ mevalonic acid), and the corresponding concentration of compounds was added for 16 hours. After compound treatment, the cells were lysed with lysis buffer (E397A, Promega), and after adding luciferase substrate (E1500, Promega), the activity of SRE-driven luciferase was measured by 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 by the above BioTek microplate reader (including but not limited to this type of instrument), and was used as an internal control. The ratio of SRE-driven luciferase activity to green fluorescent protein fluorescence intensity was used as an indicator of SREBP pathway activity. The test data of each test compound was analyzed by prism software to obtain the IC 50 parameter.

[0698] Biological test example 3: real-time fluorescent quantitative PCR

[0699] Liver or liver organoid samples were homogenized in TRI Reagent (T9424, Sigma) and total RNA was extracted according to the manufacturer's protocol. Equal amounts of RNA templates were used to synthesize cDNA with oligodT primers and MLV reverse transcriptase (Promega). Quantitative data of genes were collected by Bio-Rad CFX96 real-time PCR System and the relative amount of mRNA of genes was quantified by the relative CT method. The primer sequences used are shown in the following table:

[0700]

[0701] Measurement of serum and liver metabolic parameters

[0702] After drug treatment of mice, the mice were sacrificed after removing the feed for 4 hours. Blood and liver were collected. After blood coagulation, centrifugation at 1500g for 10 minutes at 4°C, the supernatant was the serum. The lipids in the liver were extracted by the method of chloroform / methanol, first homogenized by homogenizer Precelly24, centrifuged at 16000g for 10 minutes at 4°C, the organic phase was transferred to a new tube, dried by nitrogen, then dissolved with ethanol. The total cholesterol and triglyceride levels in blood and liver were measured by cholesterol and triglyceride kits (Shanghai Kewei Bioengineering Co., Ltd.), respectively. ALT (LE-M0477, Lai Er Bio-tech), AST (LE-M0568, Lai Er Bio-tech) in serum were measured according to the corresponding manufacturer's manual, using the analysis system of Hisun Medical Electronics (Shanghai) Co., Ltd.

[0703] Biological test example 5: analysis of liver tissue sections

[0704] Hematoxylin-eosin staining: The liver after removal was fixed in 4% paraformaldehyde at 4°C, paraffin-embedded, sectioned with a paraffin microtome (Leica RM2235) at 7 pm thickness, deparaffinated and rehydrated, and then stained with hematoxylin-eosin staining kit (6765001, 6766010, Thermo Scientific). Images were taken with an Olympus VS 120 slide microscope, and quantified with ImageJ software. Oil Red O staining: The liver was embedded with OCT embedding medium (Leica), sectioned into 7 pm thickness with a freezing microtome (Leica CM1950), and stained with Oil Red O (O0625, Sigma). Images were taken with an Olympus VS 120 slide microscope, and quantified with ImageJ software. Sirius Red collagen staining: Paraffin sections of the liver were deparaffinated and rehydrated, and then stained with Sirius Red staining kit (ab150681, Abeam) according to the manufacturer’s instructions. Immunofluorescence staining: The liver was embedded with OCT embedding medium (Leica), sectioned into 7 pm thickness with a freezing microtome (Leica CM1950), and 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) for specific staining of the nucleus. After mounting, images were taken with a spinning disk confocal microscope (Nikon CSU-W1 SoRa) equipped with a polarized light filter, and quantified with ImageJ software.

[0705] Mouse arterial tree isolation and atherosclerotic plaque Sudan IV staining

[0706] After the end of the mouse dosing experiment, the aortas were isolated and fixed in 4% PFA, and after removing the perivascular adipose tissue with an ophthalmic forceps under a stereomicroscope, the atherosclerotic plaques were stained with Sudan IV and washed with 70% ethanol. After staining, the aortic tree was imaged with a ZEISS Axio Zoom.V16 stereomicroscope. Atherosclerotic lesions were quantified with ImageJ software.

[0707] Mouse liver organoid preparation and culture

[0708] 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 a 1:3 mixture of culture medium and basement membrane extract (BME) (R&D Systems, 3533-010-02). 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. After 24 hours, using DMSO as a blank control, the medium was changed to contain different concentrations of OCA or 25-HL. 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 given drug treatment for 72 hours.

[0709] Biological Testing Example 8: Histochemical Staining of Mouse Liver Organoids

[0710] Immunocytochemistry: The organoids were fixed in immunostaining fixative (Beyotime Biotechnology, P0098) at 4°C overnight. Then washed in PBS, and then treated with PBS containing 0.5% Triton X-100 for 20 min at room temperature. Then the organoids were blocked with PBS containing 10% goat serum for 1 hour at room temperature, and incubated with primary antibody (Anti-alpha smooth muscle Actin, ab7817, Abeam, 1:500 dilution) at 4°C overnight. The next day, the organoids were washed, and co-incubated with secondary antibody, fluorescein (FITC) conjugated goat anti-mouse IgG (H+L) (Jackson Immunoresearch, 115-095-003). The nuclei were counterstained with DAPI (Sigma-Aldrich, F6057). TM

[0711] For observation of lipid droplets, the organoids were fixed with 4% paraformaldehyde (PFA) for 1 hour, and stained with 250 nmol / L Nile Red (MCE, HY-D0718) for 3 min at room temperature. The organoids were rinsed with PBS twice before imaging. The stained organoids were observed under Dragonfly high-speed confocal microscope system (Andor, Dragonfly 200).

[0712] Bioassay Example 9 Inhibition effect of compounds of the present application on SREBP pathway

[0713] The inhibition effect of compounds of the present application on SREBP pathway was tested by the method of Bioassay Example 2, and the concentration gradient of each compound was designed as 0.01, 0.03, 0.1, 0.3, 1.0, 3.0, 10 μM, and the control was solvent DMSO. The IC 50 values of some compounds are shown in Table 1.

[0714] Table 1: Activity data of compounds of some examples

[0715]

[0716]

[0717]

[0718]

[0719]

[0720]

[0721] ​

[0722]

[0723]

[0724]

[0725]

[0726]

[0727]

[0728]

[0729]

[0730]

[0731]

[0732]

[0733]

[0734] Bioassay Example 10 25-Hydroxy lanosterol reduces blood lipid level, alleviates fatty liver and liver injury

[0735] It is known that elevated blood lipid level and liver lipid accumulation are high risk factors for fatty liver, and the present application further analyzes whether 25-hydroxy lanosterol can alleviate the typical symptoms of diet-induced fatty liver in mice: lipid accumulation, liver injury, inflammation and fibrosis.

[0736] Male C57BL / 6J mice and male Ldlr - / - mice (T001464, Jiangsu Jicui Yekang Biotechnology Co., Ltd.) were grouped and fed with different feeds and given different drug treatments, and the mice were randomly divided into 4 groups, 8-9 mice in each group: group 1 was C57BL / 6J wild-type mice fed with chow diet (CD) vehicle control group. The other 3 groups were Ldlr - / - knockout mice, respectively, as the chow diet (CD) vehicle control group, the AMLN diet (containing 20% fat, 22% fructose, 2% cholesterol) vehicle control group, and the AMLN diet 25-hydroxy lanosterol administration group (the administration concentration of 25-hydroxy lanosterol was 30 mg / kg / day). The 4 groups of mice were given intragastric administration once a day, and the food intake and body weight changes of the mice in different treatment groups were counted during the period. After 8 weeks, the blood and liver of the mice were collected, and the blood lipid, liver lipid and liver injury phenotypes were analyzed.

[0737] Results are shown in Figures 1-8 , wherein, Figure 1 is the statistical results of the weight of each group of mice per week, Figure 2 is the cumulative statistics of the food intake of each group of mice, Figure 3 is the content of total cholesterol in the blood of mice reduced by 25-hydroxy lanosterol, Figure 4 is the content of total triglyceride in the blood of mice reduced by 25-hydroxy lanosterol, Figure 5 is the content of total cholesterol in the liver of mice reduced by 25-hydroxy lanosterol, Figure 6 is the content of total triglyceride in the liver of mice reduced by 25-hydroxy lanosterol, Figure 7 is the level of aspartate aminotransferase (AST) in the blood of mice reduced by 25-hydroxy lanosterol, Figure 8 is the level of alanine aminotransferase (ALT) in the blood of mice reduced by 25-hydroxy lanosterol.

[0738] Figure 1 and Figure 2 , P value is calculated by statistical two-way ANOVA (Dunnett's multiple comparisons test) variance analysis; * indicates P<0.05; ns indicates no statistical difference. Figures 3-8 , the data is expressed as mean ± standard deviation. P value is calculated by one-way ANOVA variance analysis * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001.

[0739] The results show that after 8 weeks of continuous administration, Ldlr - / - The body weight of mice in the 25-hydroxy lanosterol group fed with AMLN feed was significantly lower than that of the AMLN feed vehicle control group. It shows that 25-hydroxy lanosterol has a good inhibitory effect on the weight gain induced by AMLN diet. The pathological features of fatty liver disease mainly include liver steatosis, liver damage, inflammatory infiltration and fibrosis. After 8 weeks of administration of the 4 groups of mice, we first detected the changes of total cholesterol, total triglyceride and other lipid levels in the blood of mice and the changes of total cholesterol and total triglyceride lipid levels in the liver. As shown in Figures 3-4 , compared with the control group, 25-hydroxy lanosterol significantly reduces the total cholesterol and total triglyceride levels in serum. At the same time, as shown in Figures 5-6 , 25-hydroxy lanosterol significantly reduces the total cholesterol and total triglyceride levels in the liver of mice. It shows that 25-hydroxy lanosterol has a good effect of reducing blood lipids and liver fat. From Figures 7-8As can be seen from Table 1, the liver injury markers, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum were significantly reduced by 25-hydroxy lanosterol. This indicates that 25-hydroxy lanosterol has a good effect of improving liver injury.

[0740] Bioassay Example 11 25-hydroxy lanosterol reduces fatty liver and atherosclerosis

[0741] Further, we performed various staining or immunohistochemistry on liver tissue sections of each group of mice to analyze the changes in liver lipid accumulation and fatty liver phenotype. The results are shown in Table 2, wherein, Figures 9-29 Figures 9-13 Table 2. HE staining and NAFLD activity score quantification of mouse liver sections: 25-hydroxy lanosterol reduces liver steatosis and significantly reduces NAFLD activity score; Figures 14-18 Table 3. Oil red O staining and quantification of mouse liver sections: 25-hydroxy lanosterol reduces lipid droplets in the liver; Figures 19-23 Table 4. Sirius red staining and quantification of mouse liver sections: 25-hydroxy lanosterol reduces fibrosis in the liver; Figures 24-29 Table 5. F4 / 80 immunostaining and quantification of mouse liver sections, and polarized light imaging indicating cholesterol crystal imaging and quantification: 25-hydroxy lanosterol reduces Kupffer cell aggregation and cholesterol crystal formation in the liver.

[0742] Figures 9-29 Quantitative analysis was performed using Image J software, and the data are expressed as mean ± standard deviation. P values were calculated using one-way ANOVA, *P < 0.05, **P < 0.01, and ***P < 0.001.

[0743] Results are shown in Table 6. Figures 9-13 Hematoxylin-eosin staining shows that 25-hydroxy lanosterol reduces the accumulation of lipid droplets in the liver of male Ldlr - / - The liver of AMLN-fed mice contained obvious large vacuolar lipid droplets and bubble-shaped degenerative hepatocytes after 8 weeks of feeding. Compared with the control group, 25-hydroxy lanosterol significantly reduced the phenotype. At the same time, Figures 14-18 Oil red O staining of liver tissue sections shows that 25-hydroxy lanosterol significantly reduces the accumulation of neutral lipid droplets, including cholesterol and fatty acids, in the liver compared with the control group. These results are consistent with Figures 1-8 The results of 25-hydroxy lanosterol reducing liver lipid content are consistent with those in Table 6, indicating that 25-hydroxy lanosterol effectively improves the liver lipid accumulation phenotype induced by AMLN diet. In addition, the present application further analyzes the changes in inflammation and fibrosis phenotypes related to fatty liver in the liver. Figures 24-29 Immunofluorescence staining shows that 25-hydroxy lanosterol reduces the accumulation of Kupffer cells in the liver of male Ldlr - / - ​In mice fed an AMLN diet, F4 / 80-specific staining showed that Kupffer cells aggregated together and formed crown-like structures around cholesterol crystals. Figures 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. Figures 24-29 ).at the same time, Figures 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.

[0744] 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 Zoom V16, Germany). Quantitative analysis of atherosclerotic plaques was performed using ImageJ software.

[0745] 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.

[0746] 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.

[0747] Figures 1-8 to Figures 9-34These 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.

[0748] 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.

[0749] 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 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 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, Col1α1) were analyzed using real-time quantitative PCR.

[0750] The results are as follows Figures 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-related gene (Col1a1, αSMA) in liver organoids, and the effect was better than that of the control drug obeticholic acid at the same concentration. Figures 37-53For bright field imaging results of liver organoids, Nile red staining and quantification results and fibrosis marker aSMA immunofluorescence staining and quantification results: 25-hydroxy lanosterol reduces lipid accumulation in liver organoids while reducing expression of fibrosis marker aSMA in liver organoids.

[0751] Figure 35 , Figure 36 Data are presented as mean ± standard deviation. P values were calculated using one-way ANOVA ns indicates no statistical difference, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001. Figure 47 and Figure 53 Quantitative analysis was performed using Image J software. Data are presented as mean ± standard deviation. P values were calculated using one-way ANOVA * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

[0752] The results show that, compared to obeticholic acid which failed to inhibit the expression of lipid synthesis genes, 25-hydroxy lanosterol can significantly inhibit the expression levels of lipid synthesis related genes ( Figure 35 ) and fibrosis related genes ( Figure 36 ). At the same time, Nile red staining experiment was used to detect the accumulation of neutral lipids in organoids, as shown in Figures 37-53 The results show that 25-hydroxy lanosterol and OCA reduce lipid accumulation in organoids, and the staining data of fibrosis marker protein aSMA show that the expression of fibrosis protein aSMA is significantly reduced. Figures 35-53 The results show that 25-hydroxy lanosterol directly reduces liver lipid accumulation and fibrosis by inhibiting the expression of lipogenesis and fibrosis genes, while OCA may indirectly inhibit liver lipid accumulation by promoting lipid oxidation. These data prove that 25-hydroxy lanosterol directly targets the regulation of liver lipid synthesis pathway and fibrosis genes.

[0753] Effect Example 13: Liver microsomal metabolic stability experiment

[0754] PBS solution (100 mM), MgCl2solution (100 mM) and NADPH solution (20 mM) were configured respectively, and then dimethyl sulfoxide (DMSO) was used to configure compound and testosterone (positive control) stock solution, and methanol was used to dilute to 100 μM for sample incubation, and stored at -10 to -30 °C. 12.5 μL of rat liver microsomes (purchased from XenoTech, item 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 MgCl2solution (100 mM) were taken respectively and added to a 96-well plate, mixed and pre-incubated at 37 °C for 5 minutes; 5 μL of substrate (test compound) solution was added to start the reaction; at each set time point 0, 5, 15, 30, 45, 60 minutes (negative group was 0, 60 minutes), 50 μL of incubation sample was taken and placed in a termination plate with 100 μL of ice-cold termination solution, vortexed for 1 minute to inactivate, and stored at -60 to -90 °C for subsequent analysis. Sample analysis of the test compound and the control compound testosterone was carried out by LC-MS / MS method. Analyst software was used to integrate, calculate and process the chromatographic peaks. Both the test compound and the control compound testosterone were semi-quantitatively analyzed, and the peak area ratio was calculated. The results are shown in Table 2 below:

[0755] Table 2: Rat liver microsomal stability results of compounds of some embodiments

[0756]

[0757]

[0758] Compared with the control compound 68, the metabolic stability of the compounds of the present application in rat liver microsomes is significantly improved, which is significantly better than that of compound 68.

[0759] Preparation of key intermediates

[0760] Preparation of examples

[0761] Example 68

[0762] Preparation of compound 68 (1R, 3aR, 5aR, 7S, 9aS, 11aR)-1-[(2R)-6-hydroxy-6-methylheptan-2-yl]-3a, 6, 6, 9a, 11a-pentamethyl-2, 3, 3a, 4, 5, 5a, 6, 7, 8, 9, 9a, 10, 11, 11a-tetradecahydro-1H-cyclopenta[1, 2-a]phenanthren-7-ol

[0763]

[0764] The first step was to dissolve the starting material lanosterol (3.00 g, 7.03 mmol, 1.0 eq) in THF (tetrahydrofuran) (273 mL), add water (68 mL) and at room temperature add NBS (0.73 g, 4.08 mmol, 0.58 eq) and stir at 25 °C for 2 hrs. TLC (n-hexane:EtOAc = 5:1, molybdenum phosphorus acid) showed that the reaction had been completed. Add dichloromethane and extract three times, dry over sodium sulfate, spin dry and then chromatograph over a column (PE:EtOAc = 50:1 to 5:1) and concentrate to give (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-5-bromo-6-hydroxy-6-methylheptan-2-yl]- 3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecahydro-1H- cyclopenta[1,2-a]phenanthren-7-ol (68-1) (1.3 g, 90% purity, 35.3% yield). 1 H NMR (400 MHz, CDC13) δ 3.98 (dd, J = 21.0, 12.3 Hz, 1H), 3.27 3.19 (m, 1H), 2.14 (d, J = 6.0 Hz, 1H), 2.00 (d, J = 11.7 Hz, 5H), 1.84 1.61 (m, 6H), 1.58 (s, 9H), 1.33 (t, J = 10.8 Hz, 9H), 1.27 1.12 (m, 3H), 1.04 (d, J = 12.2 Hz, 1H), 0.99 (d, J = 7.9 Hz, 7H), 0.94 0.84 (m, 7H), 0.80 (s, 3H), 0.69 (d, J = 2.2 Hz, 3H).

[0765] Second step (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-5-bromo-6-hydroxy-6-methylheptan-2-yl]- 3a,6,6,9a,11a-pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecahydro-1H- cyclopenta[1,2-a]phenanthrol (68-1) (900 mg, 1.72 mmol, 1.0 eq) was dissolved in THF (tetrahydrofuran) (90 mL), LAH (433 mg, 12.40 mmol, 7.2 eq) was added. After the addition was completed, it was warmed to 70 °C and refluxed, stirred for 2 hrs, TLC (n-hexane:EtOAc = 5:1, molybdenum phosphate) detection reaction was completed, the reaction system was poured into ice water (10 mL) to quench, extracted with DCM (dichloromethane) (30 mL x 3), dried over anhydrous sodium sulfate, rotary evaporation to give (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-6-hydroxy-6-methylheptan-2-yl]-3a,6,6,9a,11a- pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrol (68) (0.4 g, purity 90%, yield 48.7%). 1 H NMR (400 MHz, CDC13) δ 3.23 (d, J = 8.8 Hz, 1H), 2.02 (s, 4H), 1.92 (d, J = 8.4 Hz, 1H), 1.69 (d, J = 24.3 Hz, 7H), 1.58 (d, J = 11.1 Hz, 2H), 1.52 1.33 (m, 8H), 1.30 (s, 2H), 1.18 (d, J = 25.9 Hz, 8H), 1.01 (dd, J = 22.8, 9.8 Hz, 8H), 0.93 0.84 (m, 6H), 0.80 (s, 3H), 0.68 (s, 3H). 13 C NMR (101 MHz, CDC13) δ 134.38, 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

[0766] Example 146

[0767] Preparation of compound 146 (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-7-hydroxyheptan-2-yl]- 3a,6,6,9a,11a-pentamethyl-2,3,3a,5,5a,6,7,8,9,9a,11,11a-dodecahydro-1H- cyclopenta[1,2-a]phenanthren-7-ol

[0768]

[0769]

[0770] In the first step, (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-4-formylbutan-2-yl]-3a,6,6,9a,11a- pentamethyl-2,3,3a,4,5,5a,6,7,8,9,9a,10,11,11a-tetradecahydro-1H- cyclopenta[2,1-c]phenanthren-7-yl acetate (II) (500 mg, 1.1 mmol) was dissolved in tetrahydrofuran (30 mL), N2was bubbled for three times, cooled to 0 °C, potassium tert-butoxide (253 mg, 2.3 mmol) solid was added, and stirred at 0 °C for 30 min, (2-methoxy-2-oxoethylidene)phosphonic acid diethyl ester (500 mg, 1.1 mmol) was dissolved in THF (5 ml), added dropwise into the above reaction system, and allowed to warm to room temperature naturally, and stirred for 2 h. TLC (PE:EtOAc = 10:1) showed no starting material left, cooled to 0 °C, 1 M / HCl (2 mL) was added, separated, extracted with ethyl acetate twice, dried over anhydrous sodium sulfate, rotary evaporated, and columned to give (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthren-1-yl]hept-2-enoic acid methyl ester (67-1) (430 mg, 0.86 mmol, 76.65%) as a white solid. 1HNMR (399 MHz, CDC13) δ 7.03 - 6.85 (m, 1H), 5.80 (d, J = 15.6 Hz, 1H), 4.48 (dd, J = 11.7, 4.5 Hz, 1H), 3.773.62 (m, 3H), 2.25 (s, 1H), 2.121.92 (m, 8H), 1.89 (d, J = 21.3 Hz, 1H), 1.571.39 (m, 4H) 1.57 - 1.39 (m, 5H), 1.36 - 1.21 (m, 3H), 1.15 (dd, J = 19.7, 10.5 Hz, 3H), 0.98 (s, 3H), 0.87 (dd, J = 14.6, 4.9 Hz, 12H), 0.66 (s, 3H)

[0771] The second step, (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- tetradecahydro-1H-cyclopenta[1,2-a]phenanthren-1-yl]hept-2-enoic acid methyl ester (67-1) (380 mg, 0.762 mmol) was dissolved in ethyl acetate (380 mL), Pd / C (palladium on carbon) (38 mg, 0.762 mmol) was added, hydrogen gas was exchanged three times, stirred at room temperature for 30 min. TLC (PE:EtOAc = 10:1) no starting material left, spot test reaction was complete, filtered through celite, rinsed with ethyl acetate, spin dried, later column chromatography (PE:EtOAc = 100%-20%), concentrated to give (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-tetradecahydro-1H- cyclopenta[1,2-a]phenanthren-1-yl]heptanoic acid methyl ester (67-2) (350 mg, 0.629 mmol, 82.56%). 1 H NMR (400 MHz, CDC13) δ 4.49 (dd, J = 11.5, 4.6 Hz, 1H), 3.66 (d, J = 1.1 Hz, 3H), 2.30 (t, J = 8.0 Hz, 2H), 2.11 - 1.94 (m, 7H), 1.94 - 1.82 (m, 1H), 1.82 - 1.46 (m, 8H), 1.46 - 1.09 (m, 8H), 0.99 (s, 3H), 0.87 (q, J = 2.7, 2.2 Hz, 12H), 0.67 (s, 3H). 13CNMR (101 MHz, CDC13) δ 134.42, 134.20, 51.46, 50.38, 49.77, 44.43, 37.78, 36.86, 36.25, 35.77, 35.24, 34.18, 30.92, 30.78, 28.18, 27.90, 26.35, 25.90, 25.39, 24.23, 24.15, 21.35, 20.97, 19.17, 18.62, 18.10, 16.52, 15.73, 1.02.

[0772] The third step, (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-tetradecahydro-1H- cyclopenta[1,2-a]phenanthren-1-yl]heptanoic acid methyl ester (67-1) (100 mg, 0.2 mmol, 1.0 eq.) was dissolved in dichloromethane (10 mL), then m-chloroperbenzoic acid (77.61 mg, 0.45 mmol, 2.3 eq.) was added under ice-bath, after the addition, the reaction was continued to stir at room temperature, TLC monitoring (PE:EA = 10:1, molybdenum phosphorus acid coloration, Rf1 = 0.5, Rf2 = 0.3), after the reaction was completed, saturated aqueous sodium bicarbonate solution (5.0 mL) was added, extracted with dichloromethane (5.0 mL*3), the organic phase was dried and concentrated to give (6R)-6-((3S,5S,10S,13R,14R,17R)-3-acetyloxy-4,4,10,13,14-pentamethyltetradecahydro-11H-8,9- epoxycyclopenta[a]phenanthren-17-yl)heptanoate methyl ester (146-1) (89 mg, 0.17 mmol, 86%) white solid crude, which was used in the next step without purification.

[0773] Fourthly, methyl (6R)-6-((3S,5S,10S,13R,14R,17R)-3-acetyloxy-4,4,10,13,14-pentamethyl- 11H-8,9-cyclooxocyclopenta[a]phenanthren-17-yl)heptanoate (146-1) (89 mg, 0.17 mmol) was dissolved in acetone (10 mL), concentrated sulfuric acid (1 d) was added at room temperature, then stirred at room temperature for 5 hours, TLC monitoring (PE:EtOAc = 10:1, molybdenum phosphorus acid coloration, Rf1= 0.3, Rf2= 0.6), after the reaction was completed, the acetone was removed by concentration, saturated aqueous sodium bicarbonate solution (10.0 mL) was added, 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 (6R)-6-[(1R,3aR,5aR,7S,9aS,11aR)-7-acetyloxy-3a,6,6,9a,11a-pentamethyl-2,3,3a,5,5a,6,7,8,9,9a,11,11a-dodecahydro-1H-cyclopenta[1,2-a]phenanthren-1-yl]heptanoate (146-2) (81 mg, 0.16 mmol, 94%). 1 H NMR (399 MHz, CDC13) δ 5.43 (s, 1H), 5.29 (d, J = 5.8 Hz, 1H), 4.49 (dd, J = 11.1, 4.8 Hz, 1H), 3.65 (s, 3H), 2.29 (t, J = 7.4 Hz, 2H), 2.18 (d, J = 17.8 Hz, 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 = 4.6 Hz, 1H), 1.42 - 1.34 (m, 4H), 1.27 (d, J = 10.2 Hz, 2H), 1.20 - 1.11 (m, 2H), 1.07 - 1.00 (m, 1H), 0.98 (s, 3H), 0.93 (s, 3H), 0.86 (d, J = 6.9 Hz, 9H), 0.53 (s, 3H). 13CNMR (101MHz, CDCl3)d174.36, 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.

[0774] In the fifth step, methyl (6R)-6-[(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]heptanoate (146-2) (50 mg, 0.1 mmol) was dissolved in tetrahydrofuran (5 mL), the mixture was purged with N2, cooled to zero degrees Celsius, and lithium aluminum hydride (41 mg, 1.0 mmol) was added. The mixture was then reacted at zero degrees Celsius for 1 H. TLC (PE∶EA=10∶1) showed no residue. Methanol was added, the mixture was filtered through diatomaceous earth, concentrated, and purified by preparative HPLC to obtain a white solid (1R,3aR,5aR,7S,9aS,11aR)-1-[(2R)-7-hydroxyhept-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,5,5a,6,7,8,9,9a,11,11a-dodecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (146) (12.8 mg, 0.03 mmol, 29.8%). 1 H NMR (399MHz, CDCl3) 5.45 (s, 1H), 5.30 (s, 1H), 3.62 (t, J=6.6Hz, 2H), 3.23 (dd, J=11.2, 4.5Hz, 1H), 2.18 (d, J=17.4Hz, 1H), 2.06 (d, J=11.3Hz, 3H), 1.97 (d, J= 13.0Hz, 2H), 1.64 (m, 4H), 1.43 (m, 2H), 1.34 (dd, J=20.2, 7.5Hz, 6H), 1.25 (d, J =11.6Hz, 3H), 1.06(m, 2H), 0.98(s, 3H), 0.96(s, 3H), 0.86(s, 9H), 0.54(s, 3H). 13C NMR (101MHz, CDCl3) 120.13, 116.29, 78.95, 77.31, 77.20, 76.99, 76.67, 63.10, 50.95, 50.29, 49.08, 43.70, 38.68, 37.79, 37.34 , 36.13, 36.09, 35.68, 32.86, 31.48, 28.12, 27.90, 27.78, 26.19, 26.14, 25.56, 22.98, 22.73, 18.48, 15.78, 15.62.LC-MS: [M-17] + =411

[0775] Example 151

[0776] Preparation of compound 151 cholester-5(6)-ene-3β,4β,25-triol

[0777]

[0778] 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-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (151-1) (53.8 g, 112.4 mmol, 92.77%), a white solid. 1HNMR (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).

[0779] 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 and stirred overnight. 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 solution at 0 °C and stirred for 1 hour at room temperature. 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. 1HNMR (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).

[0780] 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. 1HNMR (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).

[0781] 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. 1HNMR (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), 0.96-0.89 (m, 1H), 0.70 (d, J=12.6Hz, 3H).

[0782] 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∶1~20∶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%). 1HNMR (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).

[0783] 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. 1HNMR (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).

[0784] 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%), white solid, chiral configuration confirmed by two-dimensional spectroscopy, results 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).

[0785] 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 of the starting material was complete as monitored by TLC (DCM:MeOH = 20:1). 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). 13CNMR (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.

[0786] The two-dimensional spectrum of compound 151 is as follows: Figures 56-57 As shown.

[0787] Example 177

[0788] Preparation of compound 177 cholester-5(6)-ene-3β,25-diol

[0789]

[0790] Weigh acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-methoxy-6-oxoylide-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 (151-6) (77 mg, 0.17 mol, 1.0 eq.) and dissolve it in tetrahydrofuran (2 mL). After purging with nitrogen, dry ice is added. Acetone was cooled to -78°C, and methyllithium (0.54 mL, 0.87 mmol) was added. The mixture was then heated to room temperature and stirred. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 10:1). After the reaction was completed, 6 mL of water was added to quench the reaction. The mixture was extracted with 5 mL of ethyl acetate three times. The organic phase was dried, concentrated, and the crude product was subjected to column chromatography (DCM: MeOH = 300:1 to 100:1) to give cholesterol-5(6)-ene-3β,25-diol (177) (35 mg, 0.083 mmol, 47.66%) as a white solid. 1H NMR (399MHz, CDCl3) δ5.32 (s, 1H), 3.50 (s, 1H), 2.24 (m, 2H), 1.98 (dd, J=18.6, 10 .9Hz, 2H), 1.82 (d, J=10.3Hz, 3H), 1.53 (d, J=10.0Hz, 3H), 1.46 (d, J=7.2Hz, 3H), 1 .42 (d, J=10.1Hz, 3H), 1.36 (d, J=12.5Hz, 4H), 1.21 (s, 3H), 1.19 (s, 6H), 1.12 (dd, J=14.4, 7.2Hz, 2H), 1.03 (m, 4H), 0.99 (s, 3H), 0.91 (d, J=6.6Hz, 3H), 0.66 (s, 3H). 13 C NMR (100MHz, CDCl3) δ140.72, 121.68, 77.31, 77.00, 76.68, 71.76, 71.11, 56.70, 56.00, 50.05, 44.38, 42.57, 42.29, 42.25, 39.72, 37 .20, 36.46, 36.39, 35.72, 31.87, 31.85, 31.61, 29.34, 29.17, 28.70, 28.23, 24.26, 21.04, 20.72, 19.38, 18.66, 11.84.LC-MS: [M-OH] + =385.30

[0791] Example 185

[0792] Preparation of compound 185 cholesterol-3β,25-diol

[0793]

[0794] In the first step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-methoxy-6-oxoylide-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 (151-6) (80 mg, 0.180 mmol) was dissolved in tetrahydrofuran (2 mL) and methanol (1 mL), and Pd(OH)2 (20 mg, 0.142 mmol) was added. The mixture was purged with hydrogen three times, stirred at 40 °C for 48 hours, and monitored by HNMR. The reaction solution was filtered, and the filtrate was concentrated to give acetic acid-(1R,3aS,3bR,7S,9aS,9bS,11aR)-1-[(2R)-6-methoxy-6-oxoylidene-2-yl]-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (185-1) (80 mg, 0.170 mmol, 94.57%) as a white solid. 1 H NMR (399MHz, CDCl3) δ4.74-4.58 (m, 1H), 3.64 (s, 3H), 2.32-2.17 (m, 2H), 2.00 (s, 3H), 1.9 2(d, J=12.7Hz, 1H), 1.78 (d, J=9.5Hz, 2H), 1.73-1.61 (m, 3H), 1.53 (d, J=10.1Hz, 3H), 1.4 8-1.40 (m, 3H), 1.33 (dd, J=23.5, 11.4Hz, 4H), 1.24 (d, J=9.9Hz, 4H), 1.20-1.12 (m, 3H), 1 .10-1.02 (m, 3H), 0.96 (d, J = 15.7Hz, 2H), 0.89 (d, J = 6.3Hz, 3H), 0.79 (s, 3H), 0.62 (s, 3H).

[0795] In the second step, methylacetic acid-(1R,3aS,3bR,7S,9aS,9bS,11aR)-1-[(2R)-6-methoxy-6-oxoylide-2-yl]-9a,11a-dimethylhexadecyl-1H-cyclopentaenoic acid-7-yl ester (185-1) (80 mg, 0.179 mmol) was dissolved in tetrahydrofuran (3 mL), purged three times with nitrogen, cooled to -78 °C, and methyllithium (0.560 mL, 0.896 mmol) was added. The mixture was then heated to room temperature and stirred for 3 hours. The reaction mixture was monitored by TLC (petroleum ether:ethyl acetate = 10:1). The reaction solution was quenched with water (15 mL), extracted with ethyl acetate (10 mL * 3), washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain 80 mg of crude product. Column chromatography purification (DCM∶MeOH=300∶1-100∶1) yielded cholesterol-3β,25-diol (185) (20 mg, 0.040 mmol, 22.08%). 1 H NMR (399MHz, CDCl3) δ5.32 (s, 0H), 3.56 (s, 1H), 1.93 (d, J=12.9Hz, 1H), 1.75 (s, 2H), 1.70-1.60 (m, 2H), 1.55 (s, 3H), 1.43 (s, 3H), 1.35 (s, 3H), 1.28(s, 3H), 1.24(s, 3H), 1.23(s, 3H), 1.19(s, 6H), 1.11-1.01(m, 4H), 0.95 (d, J=10.9Hz, 3H), 0.89 (d, J=6.5Hz, 3H), 0.77 (s, 3H), 0.62 (s, 3H). 13 C NMR (101MHz, CDCl3) δ77.31, 77.20, 77.00, 76.68, 71.35, 71.10, 56.45, 56.16, 54.30, 44.81, 44.39, 42.58, 40.00, 38.19, 36.97, 36.39, 35.73, 35.47, 35.43, 32.06, 31.50, 30.95, 29.33, 29.18, 28.71, 28.25, 27.19, 24.19, 21.23, 20.74, 18.61, 12.31, 12.06.LC-MS: [M+Na] + =427.15

[0796] Example 187

[0797] Preparation of compound 187 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

[0798]

[0799] 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.

[0800] 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 it with anhydrous sodium sulfate, and concentrate it to obtain 60 mg of crude product (187-2). Proceed directly to the next step.

[0801] 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-fluoroN-[(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. 1H 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). 13 C 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

[0802] Example 198

[0803] Preparation of compound 198 5α-cholesterol-3β,4β,25-triol

[0804]

[0805] The first step involved weighing (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) (200 mg, 0.43 mol, 1.0 eq.) and dissolving it in acetic acid (10 mL). Platinum dioxide (40 mg, 0.18 mmol) was then added, followed by stirring at 40 °C for 16 hours. The mixture was monitored by TLC (petroleum ether:ethyl acetate = 3:1). After the reaction was completed, the reaction solution was filtered and concentrated to obtain 250 mg of crude product, which was then purified by column chromatography (petroleum ether:ethyl acetate = 20:1 to 15:1) to obtain methyl (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (198-1) (130 mg, 0.28 mmol, 65.12%) as a white solid.

[0806] 1 H NMR (399MHz, Chloroform-d) δ4.70 (ddd, J=12.2, 4.7, 3.2Hz, 1H), 3.81 (t, J=3.0Hz , 1H), 3.64 (s, 3H), 2.29-2.21 (m, 2H), 2.06 (s, 3H), 1.91 (dt, J=8.5, 4.6Hz, 2H), 1. 74 (dp, J=13.0, 5.1, 4.6Hz, 5H), 1.54-1.48 (m, 3H), 1.38-1.33 (m, 6H), 1.09 (t, J=4 .7Hz, 5H), 1.03 (s, 4H), 0.90 (d, J=6.5Hz, 4H), 0.83 (d, J=3.5Hz, 5H), 0.62 (s, 3H).

[0807] In the second step, methyl (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (198-1) (100 mg, 0.22 mmol) was weighed and dissolved in tetrahydrofuran (10 mL). The solution was cooled with petroleum ether and dry ice, and then a tetrahydrofuran solution of methyl lithium (C: 1.6 M, 2.2 mmol, 10.0 eq.) was added dropwise at low temperature. The solution was then brought to room temperature and stirred for 16 hours. The solution was detected by TLC (DCM:MeOH = 20:1). After the reaction was completed, 10 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 = 300:1 ~ 100:1) to obtain 5α-cholesterol (198) (25 mg, 0.06 mmol, 27.47%) as a white solid.

[0808] 1 H NMR (399MHz, Chloroform-d) δ3.72 (s, 1H), 3.52 (s, 1H), 1.94 (d, J = 12.4Hz, 1H), 1.73 (s, 4H), 1.68 (s, 3H), 1.53 (s, 3H), 1.42 (d, J = 11.3Hz, 3H), 1.33 ( d, J=11.6Hz, 4H), 1.26 (s, 3H), 1.19 (s, 6H), 1.09-1.02 (m, 4H), 0.99 (s, 3H) , 0.97-0.92 (m, 2H), 0.89 (d, J=6.5Hz, 3H), 0.63 (s, 3H), 0.61-0.52 (m, 1H).

[0809] 13 C NMR (101MHz, Chloroform-d) δ77.31, 76.99, 76.68, 74.80, 72.26, 71.13, 56.53, 56.13, 55.21, 50.87, 48.79, 44.38, 42.58, 39.87, 36.8 4, 36.39, 35.72, 35.46, 35.36, 32.37, 29.32, 29.16, 28.23, 25.95, 25.83, 24.18, 20.74, 20.56, 18.61, 14.65, 12.05.LCMS: [M-OH-H2O] + =385.3.

[0810] Example 203

[0811] Preparation of compound 203 cholester-5(6)-ene-3β,4α,25-triol

[0812]

[0813] 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 the mixture was stirred at room temperature. TLC (petroleum ether:ethyl acetate = 5:1) was used for monitoring. 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.

[0814] 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).

[0815] In the second step, 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) (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.

[0816] 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).

[0817] 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.

[0818] The two-dimensional spectrum of compound 203 is as follows: Figures 58-59 As shown.

[0819] Example 204

[0820] Preparation of compound 204 5α-cholesterol-3β,4a,25-triol

[0821]

[0822] In the first step, acetic acid-(1R,3aS,3bS,6S,7S,9aR,9bS,11aR)-6-hydroxy-1-[(2R)-6-hydroxyhex-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 (203-2) (64 mg, 0.14 mmol, 1.0 eq.) was dissolved in glacial acetic acid (10 mL), and platinum dioxide (6.3 mg, 0.03 mmol) was added. The mixture was then purged with hydrogen gas, and the temperature was raised to 40 °C with stirring. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, the reaction solution was filtered and concentrated to obtain crude (5R)-5-[(1R,3aS,3bS,5aR,6S,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate methyl ester (204-1) (80 mg, 0.14 mmol, 99.56%), a white solid. This crude product was added directly to the next reaction without purification.

[0823] In the second step, methyl (5R)-5-[(1R,3aS,3bS,5aR,6S,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (204-1) (60 mg, 0.14 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, 1.3 mmol, 10.0 eq.) was added dropwise at low temperature. The solution was then brought to room temperature and stirred for 16 hours. The reaction was detected by TLC (dichloromethane:M methanol = 20:1). After the reaction was completed, 10 mL of saturated sodium bicarbonate aqueous solution 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 5α-cholesterol-3β,4α,25-triol (204) (12 mg, 0.026 mmol, 19.98%) as a white solid. 1H NMR (399MHz, CDCl3) δ3.38-3.23 (m, 2H), 1.95 (d, J=12.8Hz, 1H), 1.84 (d, J=13.1Hz, 2H), 1. 78-1.73(m, 1H), 1.73-1.64(m, 2H), 1.49-1.39(m, 5H), 1.36(t, J=6.7Hz, 4H), 1.30-1.25(m, 3H), 1.24 (d, J=3.2Hz, 4H), 1.19 (s, 6H), 1.15-1.09 (m, 2H), 1.07-1.03 (m, 3H), 0.98 (d, J=1 0.6Hz, 2H), 0.89 (d, J=6.5Hz, 3H), 0.84 (s, 1H), 0.82 (s, 3H), 0.69-0.64 (m, 1H), 0.63 (s, 3H) 13 C NMR (100MHz, CDCl3+10%CD3OD) δ76.05, 75.07, 70.93, 56.30, 56.12, 54.34, 50.60, 49.67, 49.46, 49.24, 49.03, 48.82, 48.6 0, 48.39, 44.16, 42.42, 39.85, 37.07, 36.12, 35.68, 34.92, 28.15, 28.03, 24.04, 22.54, 20.85, 18.46, 11.91.LCMS: [M+Na] + =443.55.

[0824] Example 205

[0825] Preparation of the compound cholesterol-3β,5,25-triol

[0826]

[0827] In the first step, acetic acid-(6R,7S,9aR,11aR)-6-hydroxy-1-[(2R)-6-methoxy-6-oxoylide-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 (151-6) (800 mg, 1.8 mmol, 1.0 eq.) was dissolved in dichloromethane (50 mL). Under ice bath conditions, m-chloroperoxybenzoic acid (541.41 mg, 2.7 mmol, 1.5 eq.) was added, followed by stirring at room temperature for 4 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was complete, [the following was added]... Extracted with 30 mL of saturated sodium bicarbonate aqueous solution and 3 x 40 mL of dichloromethane, the organic phase was dried and concentrated. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 100:1 to 5:1) to obtain methyl (5R)-5-[(3S,6aS,6bS,9aR,9R,11aS,11bR)-3-acetoxy-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]oxetane-9-yl]hexanoate (205-1) (600 mg, 1.34 mmol, 74.54%), a white solid. 1 H NMR (399MHz, CDCl3) δ4.97-4.87 (m, 1H), 2.87 (d, J=4.4Hz, 1H), 2.31-2.17 (m, 2H), 2 .00 (d, J=6.4Hz, 3H), 1.961.86 (m, 2H), 1.77 (ddd, J=14.8, 9.3, 5.6Hz, 1H), 1.70-1.5 8(m, 2H), 1.52-1.41(m, 3H), 1.33(ddd, J=12.8, 9.8, 6.1Hz, 4H), 1.25-1.14(m, 3H), 1.04 (s, 3H), 0.98-0.92 (m, 2H), 0.88 (dd, J=6.5, 1.6Hz, 3H), 0.59 (d, J=11.7Hz, 3H).

[0828] The second step involves weighing methyl (5R)-5-[(3S,6aS,6bS,9aR,9R,11aS,11bR)-3-acetoxy-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-9-yl]hexanoate (100 mg, 0.22 mmol, 1.0 eq). Dissolved in tetrahydrofuran (10 mL), bromocresol green (2.4 mg, 0.22 mmol) and sodium cyanoborohydride (66 mg, 1.06 mmol, 5.0 eq.) were added at room temperature. Under nitrogen protection, the temperature was raised to 50 °C, and boron trifluoride diethyl ether (0.11 mmol) was added with stirring. The solution color changed from green to yellow. Sodium cyanoborohydride was added dropwise to ensure slow consumption. The mixture was heated under reflux overnight, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 5:1). After the reaction was completed, the mixture was cooled to room temperature, and 10 mL of water and 10 mL of ethyl acetate were added three times for extraction. The mixture was dried, concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate = 100:1~30:1~5:1) to obtain methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-5a-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate. 205-2 (component 1) (30 mg, 0.058 mmol, 26.88%) white solid and (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-5-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate methyl ester (205-3, component 2) (30 mg, 0.058 mmol, 26.88%). 205-2 (component 1): 1 H NMR (400MHz, CDCl3) δ4.77-4.66 (m, 1H), 3.78 (d, J=3.1Hz, 1H), 3.66 (s, 3H), 2.37- 2.18(m, 2H), 2.02(s, 3H), 1.97(d, J=12.6Hz, 1H), 1.84-1.75(m, 4H), 1.72-1.64(m, 4H), 1.52-1.45(m, 3H), 1.43-1.31(m, 4H), 1.29-1.20(m, 6H), 1.15-1.07(m, 4H), 1. 03(s, 3H), 1.01-0.95(m, 2H), 0.91(d, J=6.5Hz, 3H), 0.89-0.82(m, 1H), 0.67(s, 3H)

[0829] 205-3 (Second Component):1 H NMR (400MHz, CDCl3) δ5.15 (s, 1H), 3.65 (s, 3H), 2.28 (ddt, J=25.9, 16.3, 7.8 Hz, 2H), 2.01 (s, 3H), 1.95 (d, J=12.2Hz, 1H), 1.90-1.75 (m, 2H), 1.68 (d, J=8 .0Hz, 4H), 1.64-1.52(m, 7H), 1.48-1.33(m, 8H), 1.24(s, 10H), 1.06(q, J=9. 8Hz, 4H), 0.98 (s, 3H), 0.91 (d, J=6.4Hz, 3H), 0.88-0.79 (m, 1H), 0.63 (s, 3H).

[0830] In the third step, methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-5a-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (205-2) (30 mg, 0.06 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, 0.6 mmol, 10.0 eq.) was added dropwise at low temperature. The solution was then brought to room temperature and stirred for 16 hours. The reaction was monitored 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. The mixture was dried, concentrated, and then purified by column chromatography (dichloromethane:methanol = 300:1 to 100:1) to obtain cholesterol-3β,5,25-triol (205) (6.7 mg, 0.015 mmol, 23.06%) as a white solid.

[0831] 1H NMR (399MHz, CDCl3) δ4.08 (dd, J=10.9, 5.9Hz, 1H), 1.96 (d, J=12.4Hz, 1H), 1.83 (d, J=11.4Hz, 2H), 1.69-1.52 (m, 5H), 1.42 (dd, J=25.4, 14.2Hz, 12H), 1.30-1.21 (m, 7H), 1.19 (s, 6H), 1.06 (d, J=21.4Hz, 5H), 0.97 (s, 3H), 0.89 (d, J=6.4Hz, 3H), 0.63 (s, 3H). 13C NMR (100MHz, CDCl3) δ77.31, 76.99, 76.68, 75.32, 71.10, 67.34, 56.14, 56.13, 45.87, 44.38, 43.85, 42.70, 39.98, 38.76, 36.39, 3 5.74, 34.68, 34.36, 30.85, 30.77, 29.31, 29.19, 28.24, 25.91, 24.06, 21.32, 20.79, 18.59, 16.23, 12.11, 0.99.LCMS: [M-H2O-OH] + =385.

[0832] Example 206

[0833] Preparation of the compound cholesterol-3β,6,25-triol

[0834]

[0835]

[0836] Weigh out methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-5-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (205-3) (50 mg, 0.11 mmol) and dissolve it in tetrahydrofuran (5 mL). Cool the solution with petroleum ether and dry ice, then add a solution of methyl lithium in tetrahydrofuran (C: 1.6 M, 1.1 mmol, 10.0 eq.) dropwise at low temperature. Then raise the solution to room temperature and stir for 16 hours. Monitor the reaction 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. The mixture was dried, concentrated, and the crude product was purified by column chromatography (dichloromethane:methanol = 300:1 to 100:1) to obtain cholesterol-3β,6,25-triol (206) (8.2 mg, 0.019 mmol, 17.59%) as a white solid. 1 H NMR (399MHz, CDCl3) δ3.78 (s, 1H), 3.63 (s, 1H), 1.97 (d, J = 12.6Hz, 1H), 1.78 (d, J = 14.2Hz, 3H), 1.66 (dd, J = 23.5, 12.0Hz, 5H), 1 .50 (d, J=28.4Hz, 6H), 1.421.29 (m, 8H), 1.23 (s, 5H), 1.19 (s, 6H), 1.10 (s, 4H), 1.01 (s, 3H), 0.90 (d, J=6.2Hz, 3H), 0.67 (s, 3H). 13C NMR (101MHz, CDCl3) δ77.32, 77.00, 76.68, 72.07, 71.71, 71.12, 56.17, 56.15, 54.20, 47.33, 44.39, 42.67, 39.90, 39.59, 38.49, 3 6.40, 35.72, 35.36, 31.49, 30.35, 29.70, 29.36, 29.17, 28.21, 24.21, 21.04, 20.74, 18.64, 15.79, 12.10, 1.01.LCMS: [M-H2O-OH] + =385.

[0837] The two-dimensional spectrum of compound 206 is as follows: Figures 60-61 As shown.

[0838] Example 207

[0839] Preparation of the compound cholesterol-3β,5,6,25-tetraol

[0840]

[0841]

[0842] The first step is to weigh out methyl (5R)-5-[(3S,6aS,6bS,9aR,9R,11aS,11bR)-3-acetoxy-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]oxetane-9-yl]hexanoate (205-1). (100 mg, 0.22 mmol, 1.0 eq.) was dissolved in a mixed solvent of methanol (8 mL) and water (4.0 mL), and Tween 80 (50 mg, 0.22 mmol) was added. The system was then heated to 100 °C and stirred overnight. The reaction was monitored by TLC (dichloromethane:methanol = 10:1). After the reaction was completed, the mixture was cooled to room temperature, filtered, concentrated, diluted with 10 mL of water, and extracted with ethyl acetate (10 mL x 3). The organic phase was... The crude product obtained after drying and concentration was purified by column chromatography (dichloromethane:methanol = 100:1 to 10:1) to obtain methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-5,5a-dihydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (207-1, second component) (20 mg, 0.04 mmol). 17.33%) white solid and (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-5a-hydroxy-5-methoxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate methyl ester (207-2, component 1) (30 mg, 0.06 mmol, 25.25%) 207-1 (component 2): 1 H NMR (399MHz, CDCl3) δ5.14 (q, J=5.6Hz, 1H), 3.64 (s, 3H), 3.50 (s, 1H), 2.25 (ddd, J=9.9, 8.6, 6.7 Hz, 2H), 2.15 (dd, J=12.9, 11.3Hz, 1H), 2.00 (s, 3H), 1.96 (d, J=12.1Hz, 1H), 1.81 (dd, J=13.6, 7. 8Hz, 2H), 1.71-1.58(m, 7H), 1.57-1.44(m, 4H), 1.42-1.31(m, 5H), 1.25(dt, J=18.8, 4.2Hz, 4H), 1.16 (s, 3H), 1.06 (q, J=10.4, 10.0Hz, 4H), 0.90 (d, J=6.5Hz, 3H), 0.86-0.77 (m, 1H), 0.65 (s, 3H).

[0843] 207-2 (Component 1): 1H NMR (399MHz, CDCl3) δ5.10 (dt, J=11.0, 5.6Hz, 1H), 3.64 (s, 3H), 3.24 (s, 3H), 2.91 (d, J=2.8Hz, 1H), 2.332.12 (m, 3H), 2.00 (s, 3H), 1.94 (d, J=12.5Hz, 1H), 1.86-1.66(m, 5H), 1.61-1.49(m, 5H), 1.46-1.30(m, 7H), 1.30-1.16(m, 5H), 1 .05 (d, J=11.3Hz, 6H), 0.89 (d, J=6.4Hz, 3H), 0.85-0.74 (m, 1H), 0.63 (s, 3H).

[0844] In the second step, methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-5,5a-dihydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (207-1) (50 mg, 0.11 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, 1.1 mmol, 10.0 eq.) was added dropwise at low temperature. The solution was then brought 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. The mixture was dried, concentrated, and then purified by column chromatography (dichloromethane:methanol = 300:1 to 100:1) to obtain cholesterol-3β,5,6,25-tetraol (207) (8.2 mg, 0.017 mmol, 16.05%) as a white solid. 1 H NMR (399MHz, CD3OD) δ3.98 (dd, J=11.3, 5.6Hz, 1H), 3.43 (s, 1H), 2.11-1.96 (m, 2H), 1.83 (s, 1H), 1.71 (dd, J=19.9, 11.4Hz, 3H), 1.63-1.47 (m, 5H), 1.3 7 (dd, J=22.6, 10.3Hz, 9H), 1.27 (q, J=9.5, 7.9Hz, 5H), 1.14 (d, J=3.7Hz, 10 H), 1.06 (d, J=21.4Hz, 4H), 0.93 (d, J=6.4Hz, 3H), 0.88 (s, 1H), 0.70 (s, 3H). 13C NMR (101MHz, CD3OD) δ141.04, 75.38, 75.07, 70.05, 56.04, 48.20, 48.06, 47.99, 47.84, 47.78, 47.63, 47.56, 47.42, 47.35, 47.26, 47.13, 4 7.05, 46.92, 45.13, 43.87, 42.50, 40.07, 37.86, 35.72, 33.83, 30.18, 27.82, 27.67, 23.78, 20.86, 17.77, 15.89, 11.16.LC-MS: [M-OH-OH] + =402.

[0845] Example 208

[0846] Preparation of compound 6-methoxycholest-3β,5,25-triol

[0847]

[0848] 50 mg (0.06 mmol) of methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-5a-hydroxy-5-methoxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (207-2) was dissolved in tetrahydrofuran (5 mL). The solution was cooled with petroleum ether and dry ice, and then a tetrahydrofuran solution of methyllithium (C: 1.6 M, 0.6 mmol, 10.0 eq.) was added dropwise at low temperature. The solution was then brought to room temperature and stirred for 16 hours. The reaction was monitored by TLC (DCM:MeOH = 20:1). 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 (dichloromethane:methanol = 300:1 to 100:1) to obtain 6-methoxycholest-3β,5,25-triol (208) (8.2 mg, 0.018 mmol, 16.36%) as a white solid.

[0849] 1H NMR (399MHz, CDCl3) δ4.05 (dd, J=11.4, 5.9Hz, 1H), 3.65 (d, J=6.0Hz, 1H), 3 .26 (s, 3H), 2.93 (s, 1H), 2.10 (t, J = 12.3Hz, 1H), 1.96 (d, J = 12.9Hz, 1H), 1.8 2-1.64(m, 4H), 1.62-1.45(m, 8H), 1.41-1.30(m, 7H), 1.26(d, J=2.5Hz, 3H) , 1.19 (s, 9H), 1.07 (s, 3H), 0.90 (d, J = 6.5Hz, 3H), 0.82 (s, 2H), 0.65 (s, 3H). 13 C NMR (100MHz, CDCl3) δ85.56, 77.30, 77.16, 76.99, 76.86, 76.67, 76.30, 71 .11, 70.67, 67.68, 57.98, 57.94, 56.13, 55.94, 45.94, 44.38, 42.72, 40.9 8, 39.92, 38.47, 36.39, 35.74, 32.08, 30.87, 30.50, 29.68, 29.30, 28.22, 27.48, 24.13, 21.10, 20.78, 18.60, 16.34, 12.14, 1.01.LC-MS: [M-H2O-OH] + =415.

[0850] Example 210

[0851] Preparation of compound 2105α-cholesterol-3β,7,25-triol

[0852]

[0853] 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) (500 mg, 1.12 mmol, 1.0 eq.) was weighed and dissolved in acetone (50 mL). Cobalt acetate (9.95 mg, 0.056 mmol), N-hydroxyphthalimide (18.34 mg, 0.11 mmol), and tert-butyl hydroperoxide (405.34 mg, 4.5 mmol) were added. The mixture was then stirred at room temperature and 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 = 50:1~30:1~10:1) to obtain methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-4-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 (210-1) (240 mg, 0.47 mmol, 41.88%), a white solid. 1 H NMR (399MHz, Chloroform-d) δ5.68 (d, J=1.8Hz, 1H), 4.76-4.63 (m, 1H), 3.65 (s, 3H), 2.58-2.32(m, 3H), 2.23(dq, J=18.1, 10.4, 9.2Hz, 3H), 2.03(s, 3H), 1.97(d, J=10.9Hz, 2H), 1.66 (d, J=12.2Hz, 2H), 1.51 (s, 3H), 1.35 (d, J=16.7Hz, 3H), 1.2 3(s, 5H), 1.19(s, 3H), 1.07(q, J=8.9Hz, 4H), 0.92(d, J=6.5Hz, 3H), 0.66(s, 3H).

[0854] In the second step, methyl (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-4-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 (210-1) (64 mg, 0.14 mmol) was dissolved in methanol (15 mL), and palladium / carbon (1.49 mg, 0.014 mmol, 0.1 eq.) was added at room temperature. The mixture was then purged with hydrogen and stirred at room temperature for 18 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, palladium on carbon was removed by filtration, and the organic phase was dried and purified by column chromatography (petroleum ether: ethyl acetate = 30:1~20:1~10:1) to obtain methyl (5R)-5-[(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-4-oxoylidenehexadecyl-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (210-2) (57 mg, 0.1 mmol, 70.94%) as a white solid. 1 H NMR (400MHz, CDCl3) δ4.66 (dt, J=11.1, 5.9Hz, 1H), 3.66 (s, 3H), 2.39-2.22 (m, 4H), 2.22-2.15 ( m, 1H), 2.03 (d, J = 3.0Hz, 1H), 2.01 (s, 3H), 1.97 (d, J = 12.9Hz, 1H), 1.86 (s, 2H), 1.78 (d, J = 13.4H z, 1H), 1.71-1.58 (m, 3H), 1.55-1.44 (m, 5H), 1.39 (dd, J=11.6, 6.8Hz, 3H), 1.22 (d, J=17.8Hz, 2H ), 1.12 (d, J=6.9Hz, 1H), 1.08 (s, 5H), 1.05 (d, J=4.0Hz, 2H), 0.92 (d, J=6.5Hz, 3H), 0.64 (s, 3H).

[0855] In the third step, methyl (5R)-5-[(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-4-oxoylideceta-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (210-2) (50 mg, 0.11 mmol) was dissolved in a mixed solvent of tetrahydrofuran (8 mL) and methanol (8 mL). The mixture was cooled in an ice bath, and then sodium borohydride (8.21 mg, 0.22 mmol) was added at 0 °C. The mixture was then brought to room temperature and stirred. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, saturated sodium bicarbonate aqueous solution was added to quench the reaction, followed by extraction with ethyl acetate, separation of the organic phase, drying, and concentration to obtain methyl (5R)-5-[(1R,3aS,3bS,5aR,6S,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (210-3) (40 mg, 0.07 mmol, 63.72%) as a white solid. This unpurified solid was directly added to the next reaction step.

[0856] Step 4: Weigh (5R)-5-[(1R,3aS,3bS,5aR,6S,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (210-3) (40 mg, 0.09 mmol) and dissolve it in tetrahydrofuran (10 mL). Cool with dry ice and acetone, and add methyllithium (1.6 M, 18.9 mg) dropwise at -78 °C. 0.86 mmol), after the addition was complete, the mixture was brought to room temperature and monitored by TLC (dichloromethane:methanol = 20:1). After the reaction was complete, saturated sodium bicarbonate aqueous solution was added to quench the reaction, followed by extraction with ethyl acetate. The organic phase was separated, dried, and concentrated to obtain a crude product. The crude product was purified by column chromatography (dichloromethane:methanol = 100:1-30:1-10:1) to obtain 5α-cholesterol-3β,7,25-triol (210) (10.54 mg, 0.023 mmol, 26.44%) as a white solid. 1H NMR (399MHz, CDCl3) δ3.70-3.52(m, 1H), 2.02-1.90(m, 1H), 1.79(d, J=11.9Hz, 2H), 1.69(d , J=13.8Hz, 3H), 1.49 (s, 2H), 1.46-1.35 (m, 7H), 1.33-1.22 (m, 9H), 1.19 (s, 6H), 1.16-0.98 (m, 5H), 0.95-0.87 (m, 3H), 0.80 (d, J = 10.2Hz, 3H), 0.65 (d, J = 8.5Hz, 3H).13CNMR (100MHz, C DCl3) δ71.10, 50.55, 43.60, 42.65, 39.54, 37.69, 35.53, 34.91, 31.38.LC-MS: [M-2H2O-OH] + =367.5.

[0857] Example 211

[0858] Preparation of compound 211(1R,3aS,5aS,7S,9aS,11aR)-1-[(2R)-6-hydroxyoct-2-yl]-3a,9a,11a-trimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol

[0859]

[0860]

[0861] In the first step, cuprous iodide (355 mg, 2.98 mmol, 1.0 eq) was dissolved in THF (tetrahydrofuran) (7 mL). The reaction system was cooled to approximately -30 °C, and methyl magnesium bromide (355.35 mg, 2.980 mmol, 1.5 eq, 3 M inether) was slowly added while stirring for 20 min. Then, 2-(2-bromoethyl)oxetine (300 mg, 1.987 mmol, 0.2 eq) was slowly added, and the reaction system was stirred at 0 °C for 2 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was complete, the product was extracted with ethyl acetate (25 mL * 3), the organic phase was collected, dried with anhydrous sodium sulfate, and the organic phase was dried under vacuum to obtain the crude product. The crude product was separated and purified by rapid chromatography (petroleum ether: ethyl acetate = 92:8 to 90:10) to give a yellow oily 1-bromopent-3-ol (211-1) (200 mg, purity 90.0%, yield 54.23%). 1H NMR (400MHz, CDCl3) δ3.80-3.71 (m, 1H), 3.62-3.51 (m, 2H), 2.08-1.89 (m, 2H), 1.59-1.45 (m, 2H), 0.97 (t, J=7.4Hz, 3H).

[0862] In the second step, 1-bromopentyl-3-ol (211-1) (200 mg, 1.197 mmol, 1.0 eq) was dissolved in DCM (dichloromethane) (20 mL). The reaction system was placed in an ice-water bath and cooled to about 5 °C. Acetic anhydride (0.04 mL, 3.60 mmol, 3.0 eq), DMAP (29.25 mg, 0.24 mmol, 0.2 eq), and TEA (triethylamine) (0.5 mL, 3.60 mmol, 5.0 eq) were added sequentially to the reaction system. The reaction system was stirred at room temperature for 2 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was complete, it was extracted with methanol (20 mL). The reaction solution was washed once with saturated sodium bicarbonate (20 mL) and once with water (20 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was separated and purified by rapid chromatography (petroleum ether: ethyl acetate = 97:3 to 95:5) to give colorless oily 1-bromopent-3-yl acetate (211-2) (15 mg, purity 90%, yield 20.75%). 1 HNMR (400MHz, CDCl3) δ4.95 (dq, J=12.4, 6.2Hz, 1H), 3.45-3.28 (m, 2H), 2.12 (dt, J =7.8, 5.7Hz, 2H), 2.07 (s, 3H), 1.61 (dt, J = 13.6, 6.9Hz, 2H), 0.91 (t, J = 7.5Hz, 3H)

[0863] In the third step, 1-bromopent-3-yl acetate (211-2) (180 mg, 0.86 mmol) (200 mg, 1.20 mmol, 1.0 eq) was dissolved in acetonitrile (10 mL), and triphenylphosphine (677 mg, 2.583 mmol, 3 eq) was added. The reaction system was stirred at 90 °C for 18 h under a nitrogen atmosphere. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 10:1). After the reaction was complete, the reaction system was concentrated to obtain the crude product. The crude product was separated and purified by rapid chromatography (dichloromethane: methanol = 95:5 to 90:10) to obtain a white solid 1-(bromotriphenyl-λ5-methylphospho)pent-3-yl acetate (211-3) (132 mg, purity 90%, yield 29.27%). 1H NMR(400MHz,CDCl3)δ7.94-7.63(m,15H),4.83-4.74(m,1H),3.95-3.88(m,2H),2.06(s,3H),2.03-1.92(m,2H),1.80-1.75(m,2H),0.84(t,J=7.4Hz,3H).

[0864] In the fourth step, the raw materials 1-(bromotriphenyl-λ5-methylphospho)pent-3-yl acetate (211-3) (126 mg, 0.27 mmol, 2.0 eq), 18-crown ether-6 (35 mg, 0.13 mmol, 1.0 eq) and potassium carbonate (55 mg, 0.40 mmol, 3.0 eq) were dissolved in DCM (dichloromethane) (5 mL), and the reaction mixture was reacted at 50 °C for 1 h under a nitrogen atmosphere. A 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) (50 mg, 0.13 mmol, 1.0 eq) dissolved in dichloromethane (5 mL) was added to the reaction mixture and stirred under reflux for 48 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After cooling to room temperature, the reaction mixture was concentrated to give the crude product. The crude product was separated and purified by rapid chromatography (petroleum ether:ethyl acetate = 93:7 to 94:4) to give a white solid acetic acid-(5E,7R)-7-[(1R,3aS,7S,9aR,11aR)-7-acetoxy-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]oct-5-en-3-yl ester (211-4) (15 mg, purity 90%, yield 20.75%). 1HNMR (400MHz, CDCl3) δ5.37 (d, J=4.9Hz, 1H), 5.29-5.11 (m, 2H), 4.85-4.78 (m, 1H), 4.61 (ddd, J=15. 9, 8.9, 4.2Hz, 1H), 2.40 (ddd, J=16.1, 9.3, 1.8Hz, 1H), 2.32 (d, J=6.5Hz, 2H), 2.28-2.16 (m, 1H), 2.03 (t, J=3.3Hz, 6H), 1.97 (ddd, J=15.1, 9.2, 4.1Hz, 2H), 1.86 (d, J=10.2Hz, 2H), 1.70-1.43 (m, 10H), 1.2 6-1.07 (m, 5H), 1.03 (s, 4H), 0.97 (dd, J=6.6, 2.0Hz, 3H), 0.90 (t, J=7.4Hz, 3H), 0.71 (t, J=8.4Hz, 3H).

[0865] In the fifth step, the starting material, acetic acid-(5E,7R)-7-[(1R,3aS,7S,9aR,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]oct-5-en-3-yl ester (211-4) (20 mg, 0.04 mmol, 1.0 eq), was dissolved in ethanol (3 mL). Then, palladium / carbon (5 mg, 0.04 mmol, 45%) was added, and the reaction system was replaced with a hydrogen atmosphere. The reaction system was stirred at room temperature for 18 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1, phosphomolybdic acid plate). After the reaction was completed, the reaction solution was filtered with diatomaceous earth, and the filtrate was dried under vacuum to obtain a white solid acetic acid-(7R)-7-[(1R,3aS,7S,9aS,11aR)-7-acetoxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]octyl-3-yl ester (211-5) (15 mg, purity 85%, yield 63.22%). 1 H NMR (400MHz, CDCl3) δ4.84-4.76 (m, 1H), 4.734.63 (m, 1H), 2.05 (s, 3H), 2.02 (s, 3H), 1.95 (dd, J=9.3, 3.2Hz, 1H) , 1.85-1.77(m, 2H), 1.76-1.70(m, 1H), 1.68-0.95(m, 29H), 0.89(s, 2H), 0.88(s, 3H), 0.81(s, 3H), 0.64(s, 3H).

[0866] Step 6: Acetic acid-(7R)-7-[(1R,3aS,7S,9aS,11aR)-7-acetoxy-9a,11a-dimethylhexadecylhydro-1H-cyclopentano[1,2-a]phenanthrene-1-yl]octyl-3-yl ester (211-5) (50 mg, 0.10 mmol, 1 eq) was dissolved in THF (tetrahydrofuran) (1 mL) and MeOH (methanol) (1 mL), and 1N lithium hydroxide (0.6 mL) was added dropwise. The reaction mixture was stirred at 50 °C for 2 hours. The reaction was monitored by TLC (dichloromethane:methanol = 20:1). After the reaction was complete, the reaction solution was cooled to room temperature, acidified to pH 3-4 with 1N HCl, concentrated to remove ethanol, and extracted with EtOAc (ethyl acetate) (20 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was separated and purified by rapid chromatography (dichloromethane:methanol = 95:5 to 94:6) to give a white solid (1R,3aS,5aS,7S,9aS,11aR)-1-[(2R)-6-hydroxyoct-2-yl]-3a,9a,11a-trimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol (211) (21 mg, yield 50.43%). 1 H NMR (400MHz, CDCl3) δ3.59 (s, 1H), 3.54-3.46 (m, 1H), 1.99-1.92 (m, 1H), 1.79 (d, J=9.5Hz, 2H), 1.69 (s, 2H ), 1.61-1.53(m, 2H), 1.51-1.20(m, 20H), 1.13-0.98(m, 5H), 0.97-0.89(m, 7H), 0.80(s, 3H), 0.64(s, 3H).

[0867] Example 216

[0868] Preparation of compound 216 3β,25-dihydroxy-5α-cholesterol-7-one

[0869]

[0870] 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), 1.49-1.30 (m, 8H), 1.19 (s, 6H), 1.13-1.04 (m, 3H), 0.91 (d, J=6.5Hz, 3H), 0.66 (s, 3H).

[0871] 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 HNMR (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).

[0872] 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 HNMR (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 .561.44(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).

[0873] 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-oxoylideceta-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.

[0874] Example 217

[0875] Preparation of compound 217 24-[cyclopropyl(hydroxy)methyl]-5α-cholan-3β-ol

[0876]

[0877] 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 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). 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 and 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) δ5.375.33 (m, 1H), 3.67 (s, 3H), 3.52 (dd, J=7.7, 3.3Hz, 1H), 2.352.20 (m, 4H), 2.03-1.93 (m, 2H), 1.84 (ddd, J=1 2.5, 7.5, 4.3Hz, 3H), 1.75-1.60 (m, 4H), 1.59-1.35 (m, 9H), 1.331.04 (m, 7H), 1.03-0.99 (m, 3H), 0.93 (d, J=6.6Hz, 3H), 0.68 (s, 3H).

[0878] 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 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).

[0879] 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, it 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), 3.47-3.37 (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).

[0880] 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), 1.48-1.30 (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).

[0881] 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), it 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 purified by silica gel column chromatography (petroleum ether:ethyl acetate = 100:0 to 94:6) to give (5R)-1-cyclopropyl-5-[(1R, 3aS, 3bS, 7S, 9] (aR,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 (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), 2.882.82 (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),1.41(dd,J=12.4,6.8Hz,3H),1.27(d,J=12.0Hz,3H),1.16-1.01(m,5H),0 .960.84(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).

[0882] Step 6: (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), palladium on carbon was added, and hydrogen was used to replace the precipitate three times. The reaction was monitored by HNMR. After the reaction was complete, the diatomaceous earth was filtered through palladium on carbon, 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 HNMR (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

[0883] Example 218

[0884] Preparation of compound 218 24-(2,2,2-trifluoro-1-hydroxyethyl)-5α-cholan-3β-ol

[0885]

[0886] 24-(2,2,2-trifluoro-1-hydroxyethyl)cholan-6(5)-en-3β-ol (219) (52 mg, 0.093 mmol, 1.0 eq) was dissolved in methanol (5 mL) and tetrahydrofuran (2 mL), and palladium on carbon was added. The mixture was purged three times with hydrogen. The reaction was monitored by 1H NMR. After the reaction was complete, palladium on 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-(2,2,2-trifluoro-1-hydroxyethyl)-5α-cholan-3β-ol (218) (32 mg, 0.066 mmol, purity 94.86%, yield 89%). 1 H NMR (400MHz, CDCl3) δ3.95-3.87(m, 1H), 3.64-3.54(m, 1H), 1.96(m, 1H), 1.79(m, 2H), 1.73-1.62(m, 4H), 1.47(m, 4H), 1.43-1.32 (m, 5H), 1.30-1.19 (m, 6H), 1.15-0.96 (m, 7H), 0.92 (d, J=6.5Hz, 3H), 0.85 (m, 1H), 0.80 (s, 3H), 0.65 (s, 3H) 19 F(376MHz, CDCl3)δ-80.04.

[0887] Example 219

[0888] Preparation of compound 219 24-(2,2,2-trifluoro-1-hydroxyethyl)cholan-6(5)-en-3β-ol

[0889]

[0890] (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

[0891] Example 225

[0892] Preparation of compound 225 3β,5,25-trihydroxycholest-6-one

[0893]

[0894] In the first step, acetic acid-(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 (216-2) (280 mg, 0.63 mmol) was dissolved in dichloromethane (20 mL), and m-chloroperoxybenzoic acid (249.9 mg, 1.45 mmol) was added in portions at room temperature. The mixture was then stirred at room temperature and monitored by TLC (petroleum ether:ethyl acetate = 3:1). After the reaction was completed, the reaction solution was quenched with saturated sodium bicarbonate aqueous solution, and extracted with dichloromethane. The organic phase was dried with anhydrous sodium sulfate. The crude product obtained by concentrating the reaction solution was purified by column chromatography (petroleum ether: ethyl acetate = 30:1~10:1~5:1) to give acetic acid-(3S,6aS,6bS,9aR,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-i]oxacyclopropano[2,3-m]phenanthrene-3-yl ester (225-1) (220 mg, 0.41 mmol, 64.47%), a white solid. 1 H NMR (399MHz, Chloroform-d) δ4.92 (dd, J=10.8, 5.8Hz, 1H), 2.87 (d, J=4.3Hz, 1H), 2.19- 2.05(m, 1H), 2.01(s, 1H), 1.99(s, 3H), 1.93(d, J=16.2Hz, 1H), 1.70-1.59(m, 1H), 1.53( d, J=15.7Hz, 3H), 1.47-1.40(m, 3H), 1.38-1.30(m, 7H), 1.29-1.20(m, 4H), 1.19(s, 6H), 1.05 (s, 3H), 0.98 (s, 1H), 0.96 (s, 1H), 0.89 (d, J = 6.5Hz, 3H), 0.62 (s, 1H), 0.59 (s, 3H).

[0895] In the second step, acetic acid-(3S,6aS,6bS,9aR,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-i]oxacyclopropano[2,3-m]phenanthrene-3-yl ester (225-1) (100 mg, 0.22 mmol) was dissolved in acetone (10 mL), cooled to 0 °C in an ice bath, and chromium trioxide (81.39 mg, 0.81 mmol) was added. After the addition was complete, the temperature was raised to 30 °C, and the mixture was stirred for 2 hours. The mixture was monitored by TLC (petroleum ether:ethyl acetate = 5:1). 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), washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain crude acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-5a-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-5-oxoylhexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (225-2) (65 mg, 0.12 mmol, 53.4%). 1 H NMR (399MHz, Chloroform-d) δ5.00 (dt, J=11.5, 6.0Hz, 1H), 2.98 (s, 1H), 2.73 (t, J=12.6Hz, 1H), 2.08 (dd , J=13.0, 4.5Hz, 1H), 1.99 (s, 4H), 1.95-1.86 (m, 2H), 1.84-1.76 (m, 4H), 1.75-1.60 (m, 3H), 1.51 (d, J=12. 2Hz, 3H), 1.42 (d, J=9.6Hz, 3H), 1.37-1.29 (m, 4H), 1.26 (d, J=3.0Hz, 3H), 1.22 (d, J=6.1Hz, 5H), 1.18 (s, 6H), 1.12 (q, J=9.6Hz, 2H), 1.03 (dd, J=12.1, 6.5Hz, 2H), 0.90 (d, J=6.4Hz, 3H), 0.79 (s, 3H), 0.62 (s, 3H). 13C NMR (100MHz, Chloroform-d) δ110.00, 80.14, 71.16, 70.74, 56.24, 56.09, 44.26, 44.18, 43.10, 42.45, 41.69, 39.53, 37 .26, 36.34, 35.71, 32.29, 29.67, 29.49, 29.29, 29.13, 28.06, 26.24, 23.88, 21.35, 21.31, 20.81, 18.55, 13.85, 11.98.

[0896] In the third step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-5a-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-5-oxoylhexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (225-2) (65 mg, 0.14 mmol) was dissolved in a mixed solution of methanol (3 mL) and tetrahydrofuran (3 mL). Lithium hydroxide aqueous solution (1 M, 0.21 mL, 0.21 mmol) was added at room temperature. After the addition was complete, the mixture was stirred at room temperature and monitored by TLC (petroleum ether:ethyl acetate = 1:1). After the reaction was completed, water and dichloromethane (10 mL * 3) were added for extraction, the mixture was washed once with brine, dried over anhydrous sodium sulfate, concentrated and then subjected to column chromatography (petroleum ether: ethyl acetate = 10:1 ~ 5:1 ~ 2:1) to give 3β,5,25-trihydroxycholest-6-one (225) (26 mg, 0.06 mmol, 41.93%). 1 H NMR (400MHz, Methanol-d4) δ3.88 (dt, J=11.4, 6.1Hz, 1H), 2.74 (t, J=12.5Hz, 1H) , 2.04 (d, J=11.7Hz, 1H), 1.92 (dtd, J=25.2, 13.0, 5.0Hz, 3H), 1.81-1.59 (m, 5H), 1 .54(d, J=9.2Hz, 1H), 1.48-1.37(m, 6H), 1.33-1.21(m, 6H), 1.18(d, J=9.8Hz, 1H) , 1.15 (s, 6H), 1.12-1.00 (m, 2H), 0.94 (d, J=6.4Hz, 3H), 0.77 (s, 3H), 0.67 (s, 3H). 13C NMR (101MHz, cd3od) δ213.88, 79.75, 70.03, 66.42, 56.38, 56.06, 48.19, 47.98, 47.77, 47.55, 47.34, 47.13, 46.92, 44.30, 4 3.83, 42.89, 42.16, 41.37, 37.48, 36.30, 35.63, 35.13, 29.78, 29.55, 27.82, 23.53, 20.41, 12.92, 10.98.LCMS: [M-OH-H2O] + =399.25.

[0897] Example 231

[0898] Preparation of compound 231 cholesterol-3β,4β,7,25-tetraol

[0899]

[0900]

[0901] In a 50 mL round-bottom flask, 25-hydroxy-4β-hydroxy-3β-hydroxycholest-7-one (242) (40 mg, 0.092 mmol) was dissolved in tetrahydrofuran (2 mL). Heptahydrate, cerium chloride (86 mg, 0.23 mmol), sodium borohydride (8.7 mg, 0.23 mmol), and methanol (0.2 mL) were added at room temperature. The mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC (ethyl acetate / petroleum ether = 2 / 1) to indicate completion. After the reaction was complete, the reaction mixture was added to ice water (30 mL), extracted with ethyl acetate (15 mL × 3), washed with saturated brine (20 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 (eluent: ethyl acetate / petroleum ether = 2 / 1) to give a white solid cholesterol-3β,4β,7,25-tetraol (231) (28 mg, purity: 92.41%, yield: 64.38%). 1H NMR (400MHz, DMSO) δ4.30-4.28 (m, 1H), 4.03 (s, 1H), 4.00-3.95 (m, 2H), 3. 44-3.38(m, 1H), 3.29-3.25(m, 1H), 3.13-3.02(m, 1H), 1.92-1.54(m, 8H), 1 .41-1.27(m, 10H), 1.19-1.14(m, 4H), 1.05(s, 6H), 1.01-0.97(m, 3H), 0.94 -0.92(m, 3H), 0.88(d, J=6.4Hz, 3H), 0.84-0.80(m, 1H), 0.61-0.59(m, 3H). 13 C NMR (101MHz, DMSO) δ74.55, 73.98, 72.09, 71.90, 69.23, 56.43, 56.22, 55 .43, 53.40, 50.54, 46.16, 44.61, 43.41, 43.20, 42.37, 37.47, 37.16, 36.7 2, 35.75, 35.64, 35.02, 29.89, 29.70, 29.46, 28.83, 27.20, 26.23, 22.57, 20.91, 20.77, 19.17, 19.03, 15.28, 14.20, 12.55, 12.17.LCMS[M+H-3H2O] + =401

[0902] Example 232

[0903] Preparation of compound 232 24-(2,2,2-trifluoro-1-hydroxyethyl)-5α-cholene-3β,4β-diol

[0904]

[0905]

[0906] In the first step, compound 24-(2,2,2-trifluoro-1-hydroxyethyl)cholan-5(6)-en-3β-olhexanal (219) (89 mg, 0.20 mmol, 1 eq) was dissolved in chloroform (10 mL), and selenium dioxide (33.46 mg, 0.30 mmol, 1.50 eq) and N-methylmorpholine (60.99 mg, 0.60 mmol, 3.0 eq) were added. The reaction system was stirred at 80 °C for 18 hours. The reaction was monitored for completeness by TLC (petroleum ether:ethyl acetate = 5:1). 20 mL of water was added to the reaction system, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, and then evaporated under vacuum to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 76:24) to give a white solid 24-(2,2,2-trifluoro-1-hydroxyethyl)-5α-cholan-3β,4β-diol (232-1) (40 mg, purity 90%, yield 41%). 1 H NMR (400MHz, DMSO) δ6.04 (d, J=6.7Hz, 1H), 5.49 (s, 1H), 4.42 (d, J=6.2Hz, 1H), 4. 28(d, J=2.3Hz, 1H), 3.91-3.81(m, 2H), 3.25(d, J=9.6Hz, 1H), 2.03-1.91(m, 3H), 1.74(dd, J=22.2, 13.5Hz, 3H), 1.58-1.34(m, 12H), 1.27-1.18(m, 4H), 1.11(s, 3H ), 0.98 (d, J = 10.4Hz, 2H), 0.90 (d, J = 6.3Hz, 3H), 0.86-0.79 (m, 1H), 0.65 (s, 3H).

[0907] In the second step, the reactant 24-(2,2,2-trifluoro-1-hydroxyethyl)-5α-cholan-3β,4β-diol (232-1) (40 mg, 0.087 mmol, 1 eq) was dissolved in acetone (8 mL), and p-toluenesulfonic acid (11.61 mg, 0.061 mmol, 0.7 eq) and 4A molecular sieve were added. The reaction system was stirred at room temperature for 1 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1) to ensure complete reaction, and the reaction was stopped by quenching with saturated sodium sulfite. 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 over 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 = 82:18) to give the crude white solid (6R)-6-[(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]dioxacyclopentazo-8-yl]-1,1,1-trifluoroheptane-2-ol (232-2) (35 mg, purity 90%, yield 72%).

[0908] In the third step, compound (6R)-6-[(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]dioxacyclopentazo-8-yl]-1,1,1-trifluorohept-2-ol (232-2) (16 mg, 0.032 mmol, 1.0 eq) was dissolved in ethyl acetate (5 mL), then platinum dioxide (10 mg, 0.044 mmol, 1.0 eq) was added, and the reaction system was replaced with a hydrogen atmosphere. The reaction system was stirred at room temperature for 1 h. The reaction was monitored to be complete by TLC plate (petroleum ether:ethyl acetate = 5:1). The reaction system was filtered, the organic phase was collected and concentrated to give a crude white solid (6R)-6-[(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]dioxacyclopentazo-8-yl]-1,1,1-trifluoroheptane-2-ol (232-3) (16 mg). The crude product was used directly in the next step.

[0909] In the fourth step, the reactant (6R)-6-[(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]dioxacyclopentazo-8-yl]-1,1,1-trifluoroheptane-2-ol (232-3) (16 mg, 0.03 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL), and dilute hydrochloric acid (0.2 mL, 3 mol / L) was added. The reaction system was stirred at room temperature for 40 min. The reaction was monitored for completeness by TLC (petroleum ether:ethyl acetate = 2:1). Water (10 mL) was added to the reaction system, and the aqueous layer was extracted with ethyl acetate (3 × 25 mL). The ethyl acetate layers were combined and washed with saturated brine (3 × 10 mL). The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 73:27) to give a white solid 24-(2,2,2-trifluoro-1-hydroxyethyl)-5α-cholan-3β,4β-diol (232) (12.01 mg, purity 96.9%, yield 79.06%).

[0910] 1 H NMR (400MHz, DMSO) δ6.02 (d, J=6.8Hz, 1H), 4.29 (d, J=6.0Hz, 1H), 3.90 (d, J=3 .1Hz, 1H), 3.85 (s, 1H), 3.47 (d, J=2.4Hz, 1H), 3.27 (dd, J=6.2, 4.5Hz, 1H), 1.9 1(d, J=12.2Hz, 1H), 1.81-1.49(m, 7H), 1.44-1.15(m, 15H), 1.11-0.99(m, 4H), 0.94 (s, 3H), 0.88 (d, J=6.4Hz, 3H), 0.61 (s, 3H), 0.56 (dd, J=15.2, 7.3Hz, 1H).

[0911] 13 C NMR (101MHz, DMSO) δ124.67, 82.56, 74.28, 71.92, 56.59, 56.14, 56.00, 55.16, 42.61, 37.48, 35.57, 35.55, 35. 49, 32.67, 28.22, 26.43, 26.19, 24.29, 21.43, 20.64, 19.54, 18.95, 18.90, 15.18, 12.34, 0.57.LCMS[M+H-2H2O]+ =425.5

[0912] Example 233

[0913] Preparation of compound 233 24-[cyclopropyl(hydroxy)methyl]-5α-cholan-3β,4β-diol

[0914]

[0915]

[0916] In the first step, the reactant (5R)-5-[(1R,3aS,3bR,5aR,6R,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-3a,9a,11a-trimethylhexadecylhydro-1H-cyclopentano[1,2-a]phenanthrene-1-yl]hexanoate (198-1) (200 mg, 0.43 mmol, 1.0 eq) was dissolved in methanol (30 mL), and potassium carbonate (597.45 mg, 4.323 mmol, 10.0 eq) was added. The reaction mixture was stirred at room temperature for 30 min. The reaction was monitored for completeness by TLC (petroleum ether:ethyl acetate = 5:1). Water (10 mL) was added to the reaction mixture, and the aqueous layer was extracted with ethyl acetate (3 × 25 mL). The ethyl acetate layers were combined and washed with saturated brine (3 × 10 mL). The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 77:23 to 75:25) to give a white solid (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-dihydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (233-1) (150 mg, purity 80%, yield 66%). 1 H NMR (400MHz, CDCl3) δ3.74 (s, 1H), 3.66 (s, 3H), 3.56 (s, 1H), 2.27 (dd, J=16.8, 8.6Hz, 2H), 1.95 (d, J=12.5Hz, 1H), 1.84-1.67 (m, 8H), 1.53 (s, 4H), 1.43-1.31(m, 6H), 1.29-1.19(m, 3H), 1.08(dd, J=18.6, 8.9Hz, 5H), 1.02 (s, 3H), 0.92 (d, J=6.5Hz, 3H), 0.64 (s, 3H), 0.60 (dd, J=15.1, 7.3Hz, 1H).

[0917] In the second step, the reactant (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-dihydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (233-1) (150 mg, 0.36 mmol, 1 eq) was dissolved in acetone (8 mL), and p-toluenesulfonic acid (47.48 mg, 0.250 mmol, 0.7 eq) and 4A molecular sieve were added. The reaction system was stirred at room temperature for 1 h. The reaction was monitored for completeness by TLC (petroleum ether:ethyl acetate = 10:1), and the reaction was stopped by quenching with saturated sodium sulfite. 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 over 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 = 95:5) to give a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′:1,2]phenanthrene[7,8-d][1,3]dioxane-8-yl]hexanoate (233-2) (126 mg, purity 90%, yield 69%).

[0918] 1 H NMR (400MHz, CDCl3) δ3.98 (d, J=15.0Hz, 2H), 3.67 (s, 3H), 2.27 (dd, J=17.0, 8.5Hz, 1H), 1.95 (d, J=12.9Hz, 1H), 1.831.65 (m, 3H), 1.51 (s, 2H), 1.42 (dd, J=24.1, 13.0Hz, 3H), 1.30 (s, 1H), 1.24 (t, J=12.4Hz, 2H), 1.04 (s, 2H ), 0.92 (d, J = 6.4Hz, 2H), 0.85 (d, J = 12.5Hz, 1H), 0.66 (s, 1H), 0.59 (s, 1H).

[0919] The third step involves compound (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecano-3aH-cyclopentano[1′,2′∶1,2]phenanthrene. [7,8-d][1,3]dioxane-8-yl]hexanoate methyl ester (233-2) (158 mg, 0.34 mmol, 1.0 eq) was dissolved in diethyl ether (20 mL). The reaction system was cooled to -78°C, and diisobutylaluminum hydride (1.029 mL, 1.02 mmol, 1.0 eq, 1 mol / L) was added. The reaction system was stirred at -78°C for 30 min. The reaction was monitored for completeness by TLC (petroleum ether:ethyl acetate = 10:1). The reaction system was diluted with ethyl acetate (20 mL), the reaction was quenched with saturated sodium tartrate (20 mL), and extracted with ethyl acetate (10 mL * 3). The organic phase was collected, washed twice with 20 mL of water, dried over anhydrous sodium sulfate, and evaporated under vacuum to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 96:4 to 95:5) to give a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxane-8-yl]hexanal (233-3) (125 mg, purity 90%, yield 76%).

[0920] 1 H NMR (400MHz, CDCl3) δ9.76 (s, 1H), 4.12 (q, J=7.2Hz, 0H), 3.99 (d, J=14.3Hz, 2H), 2. 47-2.32 (m, 2H), 1.95 (d, J=12.1Hz, 1H), 1.85-1.61 (m, 4H), 1.51 (s, 3H), 1.43 (dd, J =20.7, 12.2Hz, 3H), 1.30 (s, 3H), 1.28-1.20 (m, 2H), 1.09 (dd, J = 15.8, 6.7Hz, 2H), 1 .04 (s, 3H), 1.02 (s, 1H), 0.93 (d, J = 6.5Hz, 3H), 0.66 (s, 3H), 0.61 (d, J = 9.0Hz, 1H).

[0921] In the fourth step, compound (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′∶1,2]phenanthro[7,8-d][1,3]dioxane-8-yl]hexanal (233-3) (50 mg, 0.116 mmol, 1.0 eq) was dissolved in tetrahydrofuran (8 mL). After cooling the reaction system to 0 °C, cyclopropylmagnesium bromide (0.580 mL, 0.580 mmol, 5 eq) was slowly added. The reaction system was stirred at room temperature for 1 h. The reaction was monitored for completeness by TLC plate (petroleum ether: ethyl acetate = 5:1). 20 mL of water 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 = 92:8) to give a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′:1,2]phenanthrene[7,8-d][1,3]dioxane-8-yl]-1-cyclopropylhex-1-ol (233-4) (49 mg, purity 90%, yield 80%). 1 H NMR (400MHz, CDCl3) δ4.04-3.93 (m, 2H), 2.89-2.82 (m, 1H), 1.96 (d, J=12.4Hz, 1H), 1.84- 1.73 (m, 2H), 1.66 (t, J=13.9Hz, 1H), 1.51 (s, 2H), 1.42 (ddd, J=25.8, 16.4, 9.2Hz, 3H), 1.3 1(s, 1H), 1.29-1.20(m, 4H), 1.14-1.06(m, 2H), 1.04(s, 2H), 0.91(d, J=6.5Hz, 2H), 0.66(s , 1H), 0.61 (dd, J=15.3, 7.5Hz, 1H), 0.56-0.44 (m, 1H), 0.24 (dtd, J=13.1, 9.0, 4.4Hz, 1H).

[0922] In the fifth step, the reactant (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′∶1,2]phenanthro[7,8-d][1,3]dioxane-8-yl]-1-cyclopropylhex-1-ol (233-4) (25 mg, 0.05 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL), and dilute hydrochloric acid (0.2 mL, 3 mol / L) was added. The reaction system was stirred at room temperature for 40 min. The reaction was monitored for completeness by TLC plate (petroleum ether:ethyl acetate = 2:1). Water (10 mL) was added to the reaction system, and the aqueous layer was extracted with ethyl acetate (3 × 25 mL). The EA layers were combined and washed with saturated brine (3 × 10 mL). The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 73:27) to give a white solid 24-[cyclopropyl(hydroxy)methyl]-5α-cholan-3β,4β-diol (233) (3.38 mg, purity 95.09%, yield 14.17%).

[0923] 1 H NMR (400MHz, DMSO) δ4.32-4.25 (m, 2H), 3.90 (d, J=2.6Hz, 1H), 3.47 (s, 1H), 3.29-3. 27 (m, 1H), 2.78 (dd, J=7.3, 4.0Hz, 1H), 1.91 (d, J=12.4Hz, 1H), 1.74 (d, J=15.2Hz, 1 H), 1.69-1.53(m, 5H), 1.40-1.22(m, 17H), 1.08-0.98(m, 4H), 0.94(s, 3H), 0.87(d, J=6.5Hz, 3H), 0.61 (s, 3H), 0.58-0.50 (m, 1H), 0.36-0.26 (m, 2H), 0.23-0.07 (m, 2H). 13 CNMR (101MHz, DMSO) δ74.28, 73.66, 71.92, 56.60, 56.19, 55.16, 48.91, 35.57, 35.55, 32.53, 30.29, 28.28, 26.44, 26.19, 24.29, 22.22, 20.64, 20.51, 18.99, 18.97, 18.08, 12.30, 0.64.LCMS[M+H-2H2O] + =397.5

[0924] Example 234

[0925] Preparation of compound 234 24-[cyclopropyl(hydroxy)methyl]-5α-cholan-3β,4β-diol

[0926]

[0927]

[0928] In the first step, compound 2-bromo-1-fluorobenzene (163 mg, 0.93 mmol, 10 eq) 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, n-butyllithium and 2.5 M n-hexane solution (0.41 mL, 1.02 mmol, 11 eq) were added dropwise using a syringe. The reaction was maintained at this temperature for 1 hour. Compound (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-2, 2,5a,7a-Tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxacyclopentanthro-8-yl]hexanal (233-3) (40 mg, 0.093 mmol, 1.0 eq) was dissolved in tetrahydrofuran (1 mL) and added to the above system. The reaction was incubated at room temperature for 30 minutes, and then allowed to rise naturally to room temperature. The reaction was monitored for completeness by TLC (petroleum ether:ethyl acetate = 5:1). 20 mL of ice water was added to the reaction system, and the mixture was extracted with ethyl acetate (10 mL × 3). The combined organic phases were washed with saturated brine (30 mL), dried, and concentrated to obtain the crude product. The crude product was subjected to rapid column chromatography (petroleum ether:ethyl acetate = 92:8) to give a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxacyclopentanthro-8-yl]-1-(2-fluorophenyl)hex-1-ol (234-1) (30 mg, purity: 80%, yield: 49%). 1H NMR (400MHz, CDCl3) δ7.45 (d, J=7.3Hz, 1H), 7.23 (d, J=7.0Hz, 1H), 7.15 (t, J=7.4Hz, 1H), 7.02 (m, 1H), 5.00 (d, J=7.0Hz, 1H), 3.98 (d, J=14.3Hz, 2H ), 1.75 (m, 5H), 1.67 (d, J = 15.3Hz, 3H), 1.51 (s, 4H), 1.41 (dd, J = 23.4, 11. 9Hz, 7H), 1.28 (m, 12H), 1.05 (d, J=13.5Hz, 6H), 0.88 (m, 5H), 0.64 (s, 3H).

[0929] In the second step, the reactant (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxacyclopentaman-8-yl]-1-(2-fluorophenyl)hex-1-ol (234-1) (25 mg, 0.05 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL), and dilute hydrochloric acid (0.3 mL, 3 mol / L) was added. The reaction system was stirred at room temperature for 60 min. The reaction was monitored for completeness by TLC plate (petroleum ether:ethyl acetate = 2:1). Water (10 mL) was added to the reaction system, and the aqueous layer was extracted with ethyl acetate (3 × 25 mL). The ethyl acetate layers were combined and washed with saturated brine (10 mL). The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 73:27) to give a white solid 24-[cyclopropyl(hydroxy)methyl]-5α-cholan-3β,4β-diol (234) (19.5 mg, purity: 93.21%, yield: 78.69%). 1H NMR (400MHz, CDCl3) δ7.47 (t, J=7.5Hz, 1H), 7.25 (m, 1H), 7.17 (t, J=7.4Hz, 1H), 7.09 (m, 1H ), 5.22 (s, 1H), 4.79 (s, 1H), 4.30 (d, J = 5.2Hz, 1H), 3.91 (s, 1H), 3.46 (s, 1H), 3.27 (m, 1H), 1.89 (d, J=12.3Hz, 1H), 1.64 (s, 3H), 1.54 (s, 5H), 1.33 (s, 6H), 1.22 (t, J=14.2Hz, 7H), 1.0 1 (d, J=9.9Hz, 3H), 0.93 (s, 3H), 0.84 (d, J=3.3Hz, 4H), 0.74 (d, J=6.2Hz, 1H), 0.59 (s, 3H). 13 C NMR (101MHz, DMSO) δ160.73, 133.32, 128.00, 124.78, 121.54, 115.12, 74.28, 71.93, 56.58, 56.15, 55.14, 49.99, 48.90, 42.60, 3 7.47, 35.55, 32.52, 29.43, 28.24, 28.23, 26.43, 26.19, 24.26, 22.04, 21.90, 20.79, 20.64, 18.87, 15.17, 12.28.LCMS[M+H-2H2O] + =451

[0930] Example 235

[0931] Preparation of compound 235, 25-hydroxy-4β-hydroxy-3β-hydroxy-5α-cholesterol-6-one

[0932]

[0933] In the first step, cholesterol-6(5)-ene-3β,4β,25-triol (151) (300 mg, 0.717 mmol) was dissolved in acetone (30 mL), and 3A molecular sieve (100 mg, 3.583 mmol) and anhydrous p-toluic acid (7.3 mg, 0.054 mmol) were added. The mixture was stirred at room temperature for 1 h, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 2:1) to ensure complete reaction. Water was added, and the mixture was extracted twice with ethyl acetate. After drying with anhydrous sodium sulfate, the extract was concentrated and column-chromatographically purified to obtain the compound (6R)-6-[(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]dioxacyclopentazo-8-yl]-2-methylhept-2-ol (235-1) (200 mg, 0.392 mmol, 54.76%) as a white solid. 1 H NMR (399MHz, Chloroform-d) δ5.78 (dd, J=4.8, 2.2Hz, 1H), 4.39 (d, J=5.7Hz, 1H), 1.98 (dd, J=15.9, 12.6Hz, 1H), 1.67-1.57 (m, 4H), 1.55 (s, 4H), 1.51 (s, 3H), 1.47-1.37(m, 5H), 1.33(s, 3H), 1.26(d, J=3.6Hz, 3H), 1.20(s, 6H), 1.15(s, 3H ), 1.08-1.00 (m, 3H), 0.92 (d, J = 6.6Hz, 3H), 0.82 (d, J = 6.7Hz, 3H), 0.68 (s, 3H).

[0934] The second step involves (6R)-6-[(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]dioxacyclopentazo-8-yl]-2-methylhept-2-ol (235-1)(50) Dissolve 49.68 mg (0.109 mmol) in anhydrous tetrahydrofuran (5 mL), under nitrogen protection, cool to 0°C, add borane tetrahydrofuran (49.68 mg, 0.654 mmol), stir at room temperature for 3 h, cool to 0°C, add sodium hydroxide (0.436 mL, 1.308 mmol), hydrogen peroxide (85%) (44.48 mg, 1.308 mmol), stir at room temperature for 16 h, and monitor the reaction for completeness by TLC (petroleum ether: ethyl acetate = 3:1). Add water, extract twice with ethyl acetate, dry to anhydrous sodium sulfate, concentrate and column chromatography to obtain (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aS, 12bR)-8-[(2R)-6-hydroxy-6-methylhept-2-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxane-12-ol (235-2) (3 mg, 0.006 mmol, 5.20%). 1 H NMR (399MHz, Chloroform-d) δ4.21 (d, J=6.1Hz, 1H), 4.16 (s, 1H), 3.82 (dd, J=10.5, 5.9Hz, 1H), 2.03-1.93 (m, 1H), 1.82-1.65 (m, 4H), 1.47 (s, 4H), 1. 43-1.33(m, 6H), 1.31(s, 5H), 1.27-1.22(m, 3H), 1.19(s, 6H), 1.13(s, 3H) , 1.06-0.99 (m, 3H), 0.90 (d, J=6.5Hz, 3H), 0.87-0.76 (m, 3H), 0.66 (s, 3H).

[0935] The third step involves (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aS, 12bR)-8-[(2R)-6-hydroxy-6-methylhept-2-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′:1,2]phenanthrene[7,8-d][1,3]di Oxacyclopentanol-12-ol (235-2) (20 mg, 0.042 mmol) was dissolved in dichloromethane (5 mL), and the mixture was cooled to 0°C under nitrogen protection. Tetrapropylammonium perruthenate (7.37 mg, 0.021 mmol), N-methylmorpholine oxide (4.94 mg, 0.042 mmol), and 3A molecular sieve (100 mg, 0.210 mmol) were added. The mixture was stirred at room temperature for 2 h, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 3:1) to ensure complete reaction. Direct filtration, concentration, column chromatography, petroleum ether:ethyl acetate (0-80%), yielded the compound (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-8-[(2R)-6-hydroxy-6-methylhept-2-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxane-12-one (235-3) (10 mg, 0.019 mmol, 45.19%). 1 H NMR (399MHz, CDCl3) δ4.40-4.22(m, 2H), 2.31-2.21(m, 2H), 2.11(d, J=9.5Hz, 1 H), 2.02 (d, J=12.5Hz, 1H), 1.89-1.79 (m, 1H), 1.72-1.55 (m, 2H), 1.50 (s, 5H), 1.42 (s, 3H), 1.36 (dd, J=19.4, 6.7H z, 5H), 1.30 (s, 3H), 1.23 (s, 3H), 1.19 (s, 6H), 1.13-0.98 (m, 4H), 0.91 (d, J=6.5Hz, 3H), 0.85 (s, 3H), 0.63 (s, 3H).

[0936] In the fourth step, (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-8-[(2R)-6-hydroxy-6-methylhept-2-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexadecylhydro-3aH-cyclopenta[1′,2′:1,2]phenanthro[7,8-d][1,3]dioxane-12-one (235-3) (3 mg, 0.006 mmol) was dissolved in methanol (5 mL), and hydrochloric acid (0.063 mL, 0.063 mmol) was added. The mixture was stirred at room temperature for 1 h, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 1:1) to ensure complete reaction. Extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography in petroleum ether:ethyl acetate (0-80%) to give compound 25-hydroxy-4β-hydroxy-3β-hydroxy-5α-cholest-6-one (235) (10 mg, 0.002 mmol, 32.77%). 1 H NMR (399MHz), CDCl3 )δ4.00 (d, J=8.8Hz, 2H), 2.33-2.24 (m, 2H), 2.21 (d, J=12.4Hz, 1H), 2.03 (d, J=12.4Hz, 1H), 1.77 (d, J=14.7Hz, 4H), 1.67-1.51 (m, 7H), 1.38 (dt, J=27.8, 12.6Hz, 9H), 1.19 (s, 3H), 1.14-0.99 (m, 4H), 0.91 (d, J=6.5Hz, 3H), 0.87 (s, 3H), 0.83 (s, 1H), 0.64 (s, 3H). 13 C NMR (100MHz, CDCl3) δ214.09, 77.30, 76.98, 76.66, 71.11, 56.89, 56.07, 44.30, 43.10, 41.07, 40.80, 39.57 , 37.15, 36.30, 35.62, 29.16, 28.08, 25.52, 23.92, 23.52, 21.27, 20.73, 18.58, 11.96, 0.99.ELSD-MS: [M+H] + =435.0.

[0937] Example 237

[0938] Preparation of compound 237(1R,3aS,5aR,7S,9aR,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-3a,6,6,9a,11a-pentamethylhexadecyl-1H-cyclopenta[1,2-a]phenanthrene-4,7-diol

[0939]

[0940] In the first 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, extracted with ethyl acetate, 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 diethyl 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), 1.24-1.15 (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.

[0941] In the second 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.

[0942] 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).

[0943] In the third step, acetic acid-(1R,3aS,5aR,7S,9aS,11aR)-4-hydroxy-1-[(2R)-6-methoxy-6-oxylidenehex-2-yl]-3a,6,6,9a,11a-pentamethyl-2,3,3a,3b,4,5,5a,6,7,8,9,9a,11,11a-tetradecanoyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (237-2) (20 mg, 0.040 mmol) was dissolved in a mixed solvent of ethyl acetate (3 mL) and acetic acid (0.3 mL). Palladium on carbon (10% Pd), containing 40-60% water (30 mg, 0.282 mmol), and platinum dioxide (3 mg, 0.013 mmol) were added. Then, hydrogen was used for purging, the temperature was raised to 80 °C, and the pressure was raised 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.

[0944] In the fourth step, acetic acid-(1R,3aS,5aR,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) (25 mg, 0.050 mmol) was weighed and dissolved in THF (3 mL). The solution was cooled in an ice bath, and methyllithium (0.464 mL, 0.743 mmol) was added dropwise. After the addition was complete, the reaction was detected by TLC (dichloromethane:methanol = 10:1) to confirm its completeness. Add saturated sodium bicarbonate aqueous solution, extract with ethyl acetate, separate the organic phase, dry and concentrate, and then purify by column chromatography (petroleum ether: ethyl acetate = 10:1 to 1:1) to obtain: (1R,3aS,5aR,7S,9aR,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-3a,6,6,9a,11a-pentamethylhexadecyl-1H-cyclopenta[1,2-a]phenanthrene-4,7-diol (237) (12 mg, 0.021 mmol, 43.04%). 1H NMR (400MHz, Methanol-d4) δ3.45 (td, J=10.4, 4.8Hz, 1H), 3.163.08 (m, 1H), 1.94-1.78 (m, 2H), 1.68-1.49 (m, 8H), 1.41 (d, J=13.4Hz, 7H), 1.27 (q, J= 12.6, 10.3Hz, 4H), 1.15 (s, 6H), 1.050.96 (m, 3H), 0.94 (d, J=4.1Hz, 6H), 0. 90 (d, J=5.3Hz, 3H), 0.87 (s, 3H), 0.86-0.82 (m, 1H), 0.79 (d, J=7.6Hz, 6H). 13 C NMR (100MHz, Methanol-d4) δ78.12, 72.24, 70.04, 52.10, 49.88, 45.64, 45.57, 43.86, 38.35, 37.32, 36.81, 36.34, 36.00, 35 .87, 31.87, 31.78, 28.14, 27.83, 27.67, 27.23, 26.91, 20.59, 19.95, 17.95, 15.39, 14.82, 13.52, 12.97.LCMS: [M-2H2O-OH] + =409.5.

[0945] Example 238

[0946] Preparation of compound 238 3β,25-dihydroxy-5α-cholesterol-4-one

[0947]

[0948]

[0949] In the first 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-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (254-1) (1.4 g, 3.261 mmol) was dissolved in chloroform (22 mL), and selenium dioxide (455.37 mg, 4.104 mmol) and N-methylmorpholine (989.44 mg, 9.782 mmol) were added. The reaction system was stirred overnight at 75 °C, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 2:1). The reaction solution was concentrated to obtain 2 g of crude product, which was purified by column chromatography (petroleum ether: ethyl acetate = 20:1 to 10:1) to give acetic acid-(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6-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 (238-1) (750 mg, 1.547 mmol, 47.43%). 1 H NMR (399MHz, Chloroform-d) δ5.68 (d, J=4.7Hz, 1H), 4.75-4.66 (m, 1H), 4.26-4.20 (m, 1H), 2.08(s, 3H), 2.06-1.97(m, 3H), 1.89-1.79(m, 3H), 1.69-1.58(m, 3H), 1.55(t, J=7.4Hz, 6H) , 1.46-1.41 (m, 4H), 1.36 (d, J = 12.0Hz, 5H), 1.25 (d, J = 12.0Hz, 4H), 1.20 (d, J = 2.1Hz, 9H), 1 .17-1.08(m, 4H), 1.08-0.98(m, 4H), 0.91(d, J=6.5Hz, 3H), 0.90-0.80(m, 2H), 0.66(s, 3H).

[0950] In the second step, acetic acid-(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6-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 (238-1) (750 mg, 1.628 mmol) was dissolved in dichloromethane (15 mL), and Dys-Martin oxidant (1380.96 mg, 3.256 mmol) was added. The mixture was stirred at room temperature for 1 hour. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 3:1). The mixture was filtered, diluted with water, extracted three times with ethyl acetate, washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain 1 g of crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 15:1 to 5:1) to give acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-6-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 (238-2) (400 mg, 0.828 mmol, 50.89%) as a pale yellow solid. 1 H NMR (399MHz, Chloroform-d) δ6.35 (dd, J=5.0, 2.6Hz, 1H), 5.19 (dd, J=12.5, 7.1Hz, 1H), 2.24-2.1 6 (m, 1H), 2.15 (s, 3H), 2.12 (d, J = 6.8Hz, 1H), 2.06-2.00 (m, 3H), 1.84 (d, J = 11.2Hz, 1H), 1.70 (dd, J=19.3, 11.2Hz, 1H), 1.53-1.44 (m, 3H), 1.43-1.37 (m, 4H), 1.32 (d, J=6.2Hz, 1H), 1.24 (s, 2H), 1. 20(s, 6H), 1.15-1.08(m, 3H), 1.08-1.00(m, 3H), 0.97(s, 3H), 0.92(d, J=6.5Hz, 3H), 0.67(s, 3H).

[0951] In the third step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(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-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (238-2) (420 mg, 0.916 mmol) was dissolved in tetrahydrofuran (20 mL) and methanol (10 mL), and palladium (palladium on carbon) (200 mg, 1.879 mmol) was added. The mixture was stirred at room temperature under hydrogen atmosphere for 1 hour, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 3:1). The filter cake was washed with ethyl acetate, and the filtrate was concentrated to obtain 420 mg of crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 10:1) to give acetic acid-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-6-oxoylidenehexadecyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (238-3) (30 mg, 0.062 mmol, 6.76%) as a white solid. 1 H NMR (399MHz, Chloroform-d) δ5.14 (dd, J=12.2, 7.2Hz, 1H), 2.18 (d, J=10.4Hz, 2H), 2.13 ( s, 3H), 1.94 (d, J = 14.8Hz, 2H), 1.89-1.78 (m, 2H), 1.72 (d, J = 12.8Hz, 2H), 1.52-1.44 (m, 3H ), 1.42 (s, 3H), 1.34 (t, J = 7.9Hz, 3H), 1.25 (d, J = 12.1Hz, 3H), 1.19 (s, 6H), 1.15-1.06 (m, 3 H), 1.02-0.93 (m, 3H), 0.90 (d, J=6.6Hz, 3H), 0.86-0.77 (m, 2H), 0.73 (s, 3H), 0.63 (s, 3H).

[0952] In the fourth step, acetic acid-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-6-oxoylideceta-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (238-3) (30 mg, 0.065 mmol) was dissolved in tetrahydrofuran (1 mL) and methanol (2 mL), and potassium carbonate (45.00 mg, 0.326 mmol) was added. The mixture was stirred at room temperature for 1 hour, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 3:1). The reaction solution was filtered, and the filtrate was directly concentrated to obtain 30 mg of crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain 3β,25-dihydroxy-5α-cholest-4-one (238) (5 mg, 0.011 mmol, 17.42%), a white solid.

[0953] 1 H NMR (400MHz, DMSO-d6) δ4.79 (d, J=5.1Hz, 1H), 4.01 (s, 2H), 2.18 (d, J=12.1Hz, 1 H), 2.08 (d, J=9.8Hz, 1H), 1.94-1.87 (m, 1H), 1.72 (d, J=9.1Hz, 2H), 1.64-1.56 ( m, 1H), 1.54-1.36 (m, 6H), 1.30-1.25 (m, 4H), 1.16-1.07 (m, 5H), 1.01 (s, 6H), 0. 98-0.88 (m, 4H), 0.85 (d, J = 6.4Hz, 3H), 0.83-0.67 (m, 3H), 0.58 (d, J = 3.8Hz, 6H). 13 C NMR (101MHz, DMSO) δ212.35, 74.51, 69.22, 56.16, 56.11, 56.05, 53.85, 44.54, 42.61, 42.55, 36.57, 35.70, 35.64 , 34.86, 32.56, 31.13, 30.38, 29.83, 29.64, 28.23, 24.19, 21.65, 20.68, 20.62, 18.94, 13.81, 12.30.LCMS: [M+H] + =419.30.

[0954] Example 239

[0955] Preparation of compound 239: Acetic acid-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexadecyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester

[0956]

[0957]

[0958] The first step involves preparing acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-acetoxy-6-methylhept-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 (254-2)(800 Cobalt acetate (2.91 mg, 0.016 mmol), N-hydroxyphthalimide (26.81 mg, 0.164 mmol), and tert-butyl hydroperoxide (592.48 mg, 6.574 mmol) were dissolved in acetone (16 mL), and then added. The reaction mixture was stirred at room temperature for 18 hours, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, water was added to the reaction solution, and the mixture was extracted three times with ethyl acetate. The organic phase was washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain 1 g of crude product. Column chromatography purification (petroleum ether:ethyl acetate = 30:1) yielded acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-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 (239-1) (450 mg, 0.854 mmol, 51.95%), a pale yellow solid. 1 H NMR (399MHz, Chloroform-d) δ5.68 (d, J=1.8Hz, 1H), 4.76-4.62 (m, 1H), 2.56-2.34 (m, 3H), 2.21 (s, 1H), 2 .03 (s, 3H), 2.01-1.95 (m, 2H), 1.94 (d, J = 1.5Hz, 3H), 1.86 (d, J = 9.7Hz, 1H), 1.73-1.60 (m, 3H), 1.56-1.49 (m, 3H), 1.40 (s, 6H), 1.35 (d, J = 8.8Hz, 3H), 1.29 (d, J = 17.6Hz, 2H), 1.23 (t, J = 9.0Hz, 4H), 1.19 (s, 3H), 1. 16 (d, J=5.6Hz, 3H), 1.090.96 (m, 2H), 0.91 (d, J=6.5Hz, 3H), 0.82 (d, J=6.4Hz, 1H), 0.66 (d, J=2.7Hz, 3H).

[0959] In the second step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-oxoylide-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (239-1) (400 mg, 0.799 mmol) was dissolved in tetrahydrofuran (12 mL) and methanol (24 mL), and palladium on carbon (200 mg, 1.879 mmol) was added. The reaction system was stirred at room temperature under hydrogen atmosphere for 1 hour, and the reaction was monitored to be complete by TLC (petroleum ether:ethyl acetate = 5:1). The mixture was filtered, and the filtrate was concentrated to obtain 50 g of crude product. Column chromatography purification (petroleum ether:ethyl acetate = 30:1) yielded acetic acid-(1R,3aS,3bR,7S,9aS,9bS,11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-oxoylidehexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (239-2) (250 mg, 0.472 mmol, 59.14%), a pale yellow solid. 1 H NMR (399MHz, Chloroform-d) δ4.64 (d, J=11.3Hz, 1H), 2.31 (t, J=12.5Hz, 2H), 2.18 (d, J=11.9Hz, 1H), 2.00 (s , 3H), 1.94 (d, J=1.9Hz, 3H), 1.87 (d, J=9.9Hz, 2H), 1.77 (d, J=13.8Hz, 1H), 1.70 (s, 1H), 1.64-1.59 (m, 2H), 1. 50 (d, J=11.7Hz, 3H), 1.45 (d, J=10.6Hz, 1H), 1.40 (s, 6H), 1.35 (d, J=7.2Hz, 3H), 1.23 (s, 3H), 1.19-1.09 (m, 2H), 1.08 (s, 3H), 1.02 (d, J=17.7Hz, 2H), 0.89 (d, J=6.3Hz, 2H), 0.81 (d, J=6.7Hz, 1H), 0.63 (d, J=2.6Hz, 3H).

[0960] In the third step, acetic acid-(1R,3aS,3bR,7S,9aS,9bS,11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-oxoylidehexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (239-2) (250 mg, 0.497 mmol) was dissolved in bis(2-methoxyethyl)aminosulfur trifluoride (10 mL), heated to 80 °C and stirred for 2 hours. The mixture was then monitored by TLC (petroleum ether:ethyl acetate = 10:1) to ensure complete dissolution. The reaction solution was cooled to room temperature, diluted with ethyl acetate, and quenched in water. It was extracted three times with ethyl acetate, washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain 300 mg of crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 100:1) to give acetic acid-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-4,4-difluoro-9a,11a-dimethyl-1-[(2R)-6-methylhept-5-en-2-yl]hexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (239-3) (70 mg, 0.143 mmol, 28.78%) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d6) δ5.04 (d, J=7.1Hz, 1H), 4.55 (dt, J=11.2, 6.1Hz, 1H), 1.93 (s, 3H), 1.89 (d, J=12.6Hz, 2H), 1.75 (d, J=15.7Hz, 2H), 1.70 (s, 3H), 1.63 (d, J=7.3Hz, 2H), 1.60 (s, 3H), 1.5 2 (s, 3H), 1.44 (d, J = 22.1Hz, 2H), 1.30 (t, J = 11.7Hz, 4H), 1.21 (d, J = 11.5Hz, 2H), 1.09 (d, J = 6. 7Hz, 1H), 1.040.97 (m, 2H), 0.93 (s, 1H), 0.87 (d, J=6.6Hz, 3H), 0.79 (s, 3H), 0.63-0.53 (m, 3H).

[0961] In the fourth step, acetic acid-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-4,4-difluoro-9a,11a-dimethyl-1-[(2R)-6-methylhept-5-en-2-yl]hexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (239-3) (70 mg, 0.151 mmol) was dissolved in tetrahydrofuran (3 mL) and water (0.75 mL), and N-bromosuccinimide (26.8 mg, 0.15 mmol) was added. The mixture was stirred at room temperature for 2 hours and monitored by TLC (petroleum ether:ethyl acetate = 5:1). The reaction solution was diluted with water, extracted three times with ethyl acetate, washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain 100 mg of crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 20:1) to give acetic acid-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-1-[(2R)-5-bromo-6-hydroxy-6-methylhept-2-yl]-4,4-difluoro-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (239-4) (50 mg, 0.085 mmol, 56.15%) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d6) δ4.74 (s, 1H), 4.55 (dt, J=11.2, 5.9Hz, 1H), 3.78 (dd, J=22.2, 11.1Hz , 1H), 1.93 (s, 3H), 1.88 (s, 1H), 1.70 (s, 4H), 1.59 (d, J = 22.7Hz, 2H), 1.49 (d, J = 20.7Hz, 2H), 1.39 (d, J=13.0Hz, 3H), 1.30 (t, J=11.9Hz, 5H), 1.22 (s, 3H), 1.15 (d, J=1.5Hz, 3H), 1.09 (d, J=17.9Hz, 2H), 1.04-0.95 (m, 2H), 0.87 (t, J=7.2Hz, 3H), 0.79 (s, 3H), 0.61 (d, J=3.8Hz, 3H).

[0962] In the fifth step, acetic acid-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-1-[(2R)-5-bromo-6-hydroxy-6-methylhept-2-yl]-4,4-difluoro-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (239-4) (50 mg, 0.089 mmol) was dissolved in tetrahydrofuran (3 mL), and lithium aluminum hydride (33.79 mg, 0.890 mmol) was added. The mixture was stirred at 70 °C for 1 hour and monitored by TLC (petroleum ether:ethyl acetate = 2:1). The reaction solution was quenched with water, extracted three times with ethyl acetate, washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain 60 mg of crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 10:1) to give acetic acid-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (239) (10 mg, 0.020 mmol, 22.11%), a white solid.

[0963] 1 H NMR (400MHz, DMSO-d6) δ4.53 (d, J=5.1Hz, 1H), 4.03 (s, 1H), 3.30 (s, 1H), 1.92-1.85 (m, 1 H), 1.73 (d, J=8.5Hz, 2H), 1.67-1.54 (m, 5H), 1.50-1.38 (m, 2H), 1.35-1.29 (m, 3H), 1.25 ( s, 3H), 1.21 (d, J=9.6Hz, 4H), 1.15 (d, J=8.6Hz, 3H), 1.12-1.03 (m, 3H), 1.01 (s, 6H), 0.93 (d, J=17.4Hz, 2H), 0.86 (d, J=6.3Hz, 3H), 0.72 (d, J=20.4Hz, 3H), 0.58 (d, J=18.8Hz, 3H). 13 C NMR (100MHz, DMSO-d6) δ69.22, 55.15, 50.26, 48.68, 44.54, 43.11, 37.19, 36.60, 36.27, 35.6 0, 35.00, 31.45, 29.82, 28.41, 25.60, 21.16, 20.68, 19.02, 12.07, 11.48.LC-MS: [M-H2O-HF] + =403.40.

[0964] Example 240

[0965] Preparation of compound 240, cholesterol-3β,6α,25-triol

[0966]

[0967] Cholesterol-6(5)-en-3β,25-diol (177) (50 mg, 0.07 mmol) was dissolved in tetrahydrofuran (5 mL), protected with nitrogen, and cooled to -10 °C. 1 M borane dimethyl sulfide solution (0.37 mL, 0.42 mmol) was added dropwise, and the mixture was allowed to rise naturally to room temperature with stirring for 3 h. The mixture was then cooled to 0 °C in an ice bath, and sodium hydroxide (10 M, 57.6 mg, 0.84 mmol) was added, followed by hydrogen peroxide (31%, 48 mg, 0.84 mmol). The mixture was stirred at room temperature for 16 h, and the reaction was monitored for completeness by TLC (petroleum ether:ethyl acetate = 1:1). The reaction was quenched with water, washed twice with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 30:1–2:1) to obtain cholesterol-3β,6α,25-triol (240) (10 mg, 0.021 mmol, 17.23%) as a white solid. 1 H NMR (399MHz, Chloroforn-d) δ3.56 (s, 1H), 3.40 (s, 1H), 2.18 (d, J=11.5Hz, 1H), 1.96 (d, J=11.5Hz, 1H), 1.79 (s, 2H), 1.56 (s, 6H), 1. 40 (s, 7H), 1.25 (d, J = 11.1Hz, 6H), 1.19 (s, 6H), 1.03 (s, 2H), 1.03 (s, 4H), 0.90 (d, J = 6.6Hz, 2H), 0.81 (d, J = 9.5Hz, 3H), 0.63 (s, 3H). 13 C NMR (100MHz, CDCl3) δ71.22, 71.09, 69.40, 56.09, 53.74, 51.64, 43.08, 42.64, 41.64, 37 .22, 35.69, 34.25, 31.01, 29.65, 23.99, 20.95, 20.49, 18.18, 0.99.ELSD-MS: [M-H2O-OH] + =385.7.

[0968] Example 241

[0969] Preparation of compound 241, 24-(1-hydroxypropyl)-5α-cholan-3β,4β-diol

[0970]

[0971] In the first step, methyl (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-dihydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (233-1) (40 mg, 0.08 mmol, 1.0 eq) was dissolved in diethyl ether (10 mL). The reaction system was cooled to -78 °C, and diisobutylaluminum hydride (0.5 mL, 0.5 mmol, 6.0 eq, 1 mol / L) was added. The reaction system was stirred at -78 °C for 1 h. The reaction was monitored for completeness by TLC (petroleum ether:ethyl acetate = 1:1). The reaction system was diluted with ethyl acetate (20 mL), the reaction was quenched with saturated sodium tartrate (20 mL), and extracted with ethyl acetate (10 mL * 3). The organic phase was collected, washed twice with 20 mL of water, dried with anhydrous sodium sulfate, and evaporated under vacuum to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 74:26) to give a white solid (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-dihydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanal (241-1) (30 mg, purity 40%, yield 35%).

[0972] In the second step, compound (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-dihydroxy-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanal (241-1) (30 mg, 0.031 mmol, 1.0 eq) was dissolved in tetrahydrofuran (8 mL). After cooling the reaction system to 0 °C, ethyl magnesium bromide (0.077 mL, 0.154 mmol, 5 eq) was slowly added, and the reaction system was stirred at room temperature for 1 h. The reaction was confirmed to be complete by TLC (petroleum ether:ethyl acetate = 5:1). 20 mL of water 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 = 74:26) to give a white solid 24-(1-hydroxypropyl)-5α-cholan-3β,4β-diol (241) (10.55 mg, purity 93.23%, yield 76%).

[0973] 1H NMR (400MHz, DMSO) δ4.28 (d, J=6.0Hz, 1H), 4.19 (dd, J=5.2, 3.0Hz, 1H), 3.88 (d, J=2.9Hz, 1H), 3.47 (d, J=2.2Hz, 1H), 3.29-3.24 (m, 2H), 1.90 (d, J=12.7Hz, 1H), 1.70-1.54 (m, 5H), 1 .43-1.33(m, 4H), 1.30(s, 3H), 1.25(d, J=9.8Hz, 8H), 1.19(s, 3H), 1.08-0.99(m, 4H), 0.9 4 (s, 3H), 0.87 (d, J = 6.5Hz, 3H), 0.83 (t, J = 7.4Hz, 4H), 0.61 (s, 3H), 0.54 (d, J = 9.5Hz, 1H). 13 C NMR (101MHz, CDCl3) δ74.86, 73.49, 72.31, 56.57, 56.10, 55.26, 48.84, 42.61, 39.91, 37.50, 36.88, 35.50, 35.40, 32.40, 31.44, 30.20, 29.71, 28.25, 26.00, 25.87, 24.21, 22.10, 20.59, 18.65, 14.68, 12.07, 9.90.LCMS[M+H-2H2O] + =385

[0974] Example 242

[0975] Preparation of compound 242, 25-hydroxy-4β-hydroxy-3β-hydroxy-5α-cholesterol-7-one

[0976]

[0977] In the first step, cholesterol-6(5)-ene-3β,4β,25-triol (151) (363 mg, 0.87 mmol) was dissolved in a 50 mL round-bottom flask containing 10 mL of dichloromethane at room temperature. Triethylamine (702 mg, 6.94 mmol), 4-dimethylaminopyridine (106 mg, 0.87 mmol), and acetic anhydride (265 mg, 2.60 mmol) were added at room temperature. The mixture was then stirred at room temperature for 16 hours. After the reaction was complete, it was detected by TLC (petroleum ether:ethyl acetate = 8:1). The reaction was quenched with water (50 mL), extracted with ethyl acetate (30 mL × 3), and the organic phase was saturated with brine (…). Wash with 50 mL of water, dry with anhydrous sodium sulfate, concentrate the organic phase, and purify by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to give a white solid acetic acid-(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-acetoxy-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-6-yl ester (242-1) (334 mg, purity: 95%, yield: 72.79%). 1 H NMR (400MHz, CDCl3) δ5.89-5.73(m, 1H), 5.50(d, J=2.3Hz, 2H), 4.78-4.72(m ,1H),2.07(s,3H),2.05-2.01(m,1H),2.01(s,3H),1.93-1.80(m,2H),1.72- 1.64(m, 1H), 1.63-1.31(m, 15H), 1.28-1.25(m, 2H), 1.21(s, 6H), 1.13(s, 3H ), 1.09-0.96 (m, 4H), 0.93 (d, J=6.5Hz, 3H), 0.90-0.81 (m, 1H), 0.67 (s, 3H).

[0978] In the second step, acetic acid-(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-acetoxy-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-6-yl ester (242-1) (170 mg, 0.34 mmol) was dissolved in a 50 mL round-bottom flask containing 5 mL of acetone. N-hydroxyphthalimide (11 mg, 0.068 mmol), cobalt acetate (6 mg, 0.034 mmol), and tert-butyl hydroperoxide (122 mg, 1.35 mmol) were added at room temperature, followed by stirring at room temperature for 16 hours. After the reaction was complete (petroleum ether: ethyl acetate = 5:1), the mixture was quenched with water (20 mL), extracted with ethyl acetate (10 mL × 3), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give a white solid acetic acid-(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-acetoxy-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-6-yl ester (242-2) (90 mg, purity: 90%, yield: 46.36%). 1 H NMR (400MHz, CDCl3) δ5.91 (d, J=14.2Hz, 1H), 5.61 (d, J=2.4Hz, 1H), 4.80 (dt, J=12.2, 4.0Hz, 1H), 2.43-2.27 (m, 2H), 2.10 (s, 3H), 2.03 (s, 3H), 2.00-1.80 (m, 4H), 1.51-1.24 (m, 16H), 1.21 (s, 6H), 1.20-0.98 (m, 5H), 0.94 (d, J=6.5Hz, 3H), 0.68 (s, 3H). 13C NMR (101MHz, CDCl3) δ202.20, 170.06, 169.47, 158.31, 131.14, 73.61, 71.51, 71.12, 54.68, 50.80, 49.91, 45.95, 44.37, 43.06, 38.53, 37.93, 36.42, 35.80, 35.69, 29.31, 29.27, 28.52, 26.20, 22.21, 21.15, 20.99, 20.78, 18.83, 17.91, 11.92.

[0979] In the third step, in a 50 mL round-bottom flask, acetic acid-(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-acetoxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-4-oxoylide-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecano-1H-cyclopenta[1,2-i]phenanthrene-6-yl ester (242-2) (50 mg, 0.097 mmol) was dissolved in a 50 mL round-bottom flask containing tetrahydrofuran (1 mL) and methanol (1 mL). The solution was then incubated at room temperature. Potassium carbonate (67 mg, 0.48 mmol) and water (0.5 mL) were added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, it was detected by TLC (petroleum ether: ethyl acetate = 3:1). The mixture was diluted with water (20 mL), extracted with ethyl acetate (10 mL × 3), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give a white solid 2,5-hydroxy-4β-hydroxy-3β-hydroxycholest-6(5)-en-7-one (242-3) (40 mg, purity: 90%, yield: 85.99%). 1 H NMR (400MHz, CDCl3) δ5.83 (s, 1H), 4.28 (t, J=10.2Hz, 1H), 3.65 (dt, J=11.8, 4.0Hz, 1H), 2.41-2.28 (m, 2H), 2.06-1.88 (m, 5 H), 1.78-1.71 (m,, 4H), 1.58-1.35 (m, 7H), 1.37 (s, 3H), 1.21 (s, 6H), 1.19-1.00 (m, 5H), 0.94 (d, J=6.5Hz, 3H), 0.69 (s, 3H).

[0980] In the fourth step, 25-hydroxy-4β-hydroxy-3β-hydroxycholest-6(5)-en-7-one (242-3) (30 mg, 0.069 mmol) was dissolved in tetrahydrofuran (1 mL) and ethyl acetate (1 mL) in a 50 mL round-bottom flask. 10% Pd / C (30 mg) was added at room temperature, and the mixture was stirred in a hydrogen atmosphere for 3 hours at room temperature. The reaction was detected by TLC (ethyl acetate / petroleum ether = 1:1) to indicate complete reaction. After the reaction was complete, the reaction solution was diluted with ethyl acetate (20 mL), filtered through diatomaceous earth, the organic phase was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give a white solid 2,5-hydroxy-4β-hydroxy-3β-hydroxycholest-7-one (242) (23 mg, purity: 98.06%, yield: 74.78%).

[0981] 1 H NMR (400MHz, DMSO) δ4.39 (d, J=6.1Hz, 1H), 4.28 (d, J=3.5Hz, 1H), 4.03 (s, 1H), 3.43 (s, 1 H), 2.80 (t, J=13.4Hz, 1H), 2.37 (t, J=11.3Hz, 1H), 2.12-2.05 (m, 1H), 1.92-1.89 (m, 1H) ,1.83-1.76(m,2H),1.72-1.55(m,3H),1.45-1.41(m,3H),1.34-1.28(m,6H),1.25-1.23 (m, 4H), 1.19 (s, 3H), 1.04 (s, 6H), 1.00-0.91 (m, 5H), 0.88 (d, J=6.5Hz, 3H), 0.60 (s, 3H). 13 C NMR (101MHz, DMSO) δ212.65, 73.62, 71.66, 69.23, 55.58, 55.04, 50.12, 49.56, 49.40, 44.60, 44.39, 42.60, 38.81 , 36.68, 36.49, 36.06, 35.60, 29.88, 29.71, 28.49, 26.02, 25.07, 21.12, 20.74, 19.12, 14.50, 12.37.LCMS(M+Na) + =457.3

[0982] Example 244

[0983] Preparation of compound 244 cholester-3β,4β,6,25-tetraol

[0984]

[0985] Cholesterol-6(5)-en-3β,4α,25-triol (151) (30 mg, 0.072 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL), under nitrogen protection, cooled to -10 °C, and borane (0.717 mL) was added dropwise. The mixture was then allowed to rise naturally to room temperature and stirred for 3 h. Water (0.2 mL) was added dropwise at 0 °C, followed by sodium peroxyborate (28 mg, 0.717 mmol). The mixture was stirred at room temperature for 16 h, and the reaction was monitored by TLC (dichloromethane:methanol = 10:1). The reaction was then quenched with water, washed twice with ethyl acetate, dried over anhydrous sodium sulfate, evaporated to dryness, and purified by column chromatography (dichloromethane:methanol = 100%–20%) to obtain the compound cholesterol-3β,4β,6,25-tetraol (244) (10 mg, 0.005 mmol, 24%). 1 H NMR (399MHz, Chloroform-d) δ4.41 (s, 0.77H), δ4.25 (s, 0.22H), 3.94 (s, 1H), 3.59-3.50 (m, 1H), 1.82 (m, 9H), 1.66 (m, 5H) 1.37 (ddd, J=26.5, 13 .5, 6.2Hz, 8H), 1.23(s, 2H), 1.19(s, 4H), 1.14(s, 3H), 1.080.96(m, 4H) , 0.90 (d, J=6.6Hz, 2H), 0.80 (d, J=6.6Hz, 1H), 0.66 (s, 2H), 0.63 (s, 1H). 13 C NMR (101MHz, CDCl3) δ, 71.14, 56.46, 56.27, 44.37, 42.65, 35.73, 29.33, 29.12, 28.31, 25.77, 20.81, 18.63, 12.07, 1.00.ELSD-MS: [M-OH] + =419.3, [M+Na] + =459.4.

[0986] Example 245

[0987] Preparation of compound 245(1R,3aS,5aR,6R,7S,9aR,11aR)-1-[(2R)-5-(hydroxycyclopropyl)pent-2-yl]-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol

[0988]

[0989]

[0990] The compound acetic acid-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-hydroxy-1-[(2R)-6-methoxy-6-oxoylide-2-yl]-9a,11a-dimethylhexadecyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (198-1) (50 mg, 0.112 mmol) was dissolved in tetrahydrofuran (5 mL), and tetraisopropyl titanate (95.6 mg, 0.670 mmol) was added at room temperature. The mixture was purged with nitrogen three times, and ethyl magnesium bromide (1.080 mL) was added dropwise at room temperature. The mixture was stirred at room temperature for 1 h, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1) to ensure complete reaction. Extracted with water and ethyl acetate, the crude product was dried after separation of the organic phase and purified by column chromatography (petroleum ether:ethyl acetate = 60:1 to 1:1) to give (1R,3aS,5aR,6R,7S,9aR,11aR)-1-[(2R)-5-(hydroxycyclopropyl)pent-2-yl]-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol (245) (10 mg, 0.022 mmol, 20.56%) as a white solid. 1 H NMR (399MHz, Chloroform-d) δ3.72(s, 1H), 3.54(s, 1H), 1.91(s, 3H), 1.74(s, 6H), 1.37(s, 7H), 1.24(s, 5H), 1 .05 (s, 4H), 1.00 (s, 4H), 0.90 (d, J = 6.4Hz, 3H), 0.84 (d, J = 15.5Hz, 2H), 0.71 (s, 2H), 0.63 (s, 3H), 0.42 (s, 2H). 13 C NMR (100MHz, CDCl3) δ, 76.66, 72.17, 55.93, 42.57, 38.81, 36.86, 35.36, 29.50, 28.37, 25.95, 24.16, 20.28, 13.43, 0.99.LC-MS: [M-H2O-OH] + =383.4.

[0991] Example 246

[0992] Preparation of compound 246 4-(trifluoromethyl)-5α-cholesterol-3β,4,25-triol

[0993]

[0994] In the first step, acetic acid-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-9a,11a-dimethyl-6-oxoylidehexadecyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (254-5) (60 mg, 0.12 mmol) was dissolved in tetrahydrofuran (5 mL). Tetrabutylammonium fluoride (3.12 mg, 0.012 mmol) and (trifluoromethyl)trimethylsilane (339.42 mg, 2.387 mmol) were added, and the mixture was stirred at room temperature for 1 hour. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). The mixture was diluted with water, extracted three times with ethyl acetate, washed once with brine, dried over anhydrous sodium sulfate, and concentrated to give 80 mg of crude product. Column chromatography purification (petroleum ether: ethyl acetate = 50:1) yielded acetic acid-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-6-hydroxy-9a,11a-dimethyl-6-(trifluoromethyl)hexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (246-1) (40 mg, 0.066 mmol, 55.59%). 1 H NMR (400MHz, DMSO-d6) δ4.89 (s, 1H), 4.85 (d, J=5.8Hz, 1H), 1.96 (s, 1H), 1.86 (s, 3H), 1.56 (td, J=32.5, 29.8, 9.7Hz, 5H), 1.32 (s, 6H), 1.27 (d, J = 10.8Hz, 3H), 1.16-0.99 (m, 4H), 0.97 (s, 3H), 0.89 (s, 1H), 0.84 (d, J = 6.2Hz, 3H), 0.76 (s, 1H), 0.58 (s, 3H).

[0995] In the second step, acetic acid-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-6-hydroxy-9a,11a-dimethyl-6-(trifluoromethyl)hexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (246-1) (40 mg, 0.070 mmol) was dissolved in methanol (1 mL) and tetrahydrofuran (2 mL), and lithium hydroxide (1 mL, 1.000 mmol) was added. The mixture was stirred at room temperature for 18 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 5:1), and the crude product was concentrated to obtain 50 mg. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 15:1) to obtain 4-(trifluoromethyl)-5α-cholest-3β,4,25-triol (246) (25 mg, 0.049 mmol, 69.59%) as a white solid.1 H NMR (400MHz, DMSO-d6) δ4.89 (s, 1H), 4.85 (d, J = 6.1Hz, 1H), 4.00 (s, 1H), 3.52 (d, J = 7.0Hz, 1H ), 1.99-1.72 (m, 2H), 1.60 (d, J = 21.0Hz, 5H), 1.48 (d, J = 12.2Hz, 2H), 1.38 (d, J = 12.8Hz, 2H), 1 .28 (d, J=14.5Hz, 4H), 1.16 (d, J=11.3Hz, 4H), 1.09 (d, J=18.4Hz, 3H), 1.01 (s, 6H), 0.97 (s, 2 H), 0.93 (s, 3H), 0.89 (d, J=7.0Hz, 2H), 0.84 (d, J=6.4Hz, 3H), 0.81-0.68 (m, 2H), 0.58 (s, 3H). 13 C NMR (101MHz, DMSO-d6) δ69.19, 69.11, 56.32, 56.09, 55.16, 47.34, 44.54, 42.41, 36.58, 36.37 , 36.08, 35.64, 34.60, 32.14, 29.85, 29.64, 28.28, 27.51, 24.10, 20.67, 18.92, 14.50, 12.29.

[0996] 19 F NMR(376MHz, DMSO-d6)δ-70.57.ELSD-MS: [M-OH] + =471.4.

[0997] Examples 248 & 249

[0998] Preparation of cholesterol isomer 248 of compound 248

[0999] Preparation of cholesterol isomer 1 of compound 249

[1000] Compounds 248 and 249 are both isomers with a single configuration, and are one of the following isomers respectively:

[1001]

[1002]

[1003] In the first step, (1R,3aS,7S,9aR,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-tetradecano-1H-cyclopenta[1,2-a]phenanthrene-7-ol (177) (1.0 g, 2.48 mmol) was dissolved in a 100 mL round-bottom flask containing anhydrous toluene (30 mL). The mixture was heated to 110 °C, and a methanol solution of cyclohexanone (10 mL) and aluminum isopropoxide (1.7 g, 2.48 mmol) was slowly added dropwise. The mixture was then stirred at 110 °C for 3 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 5:1). After the reaction was completed, the reaction was quenched with saturated ammonium chloride (50 mL), extracted with ethyl acetate (40 mL × 3), washed with saturated brine (800 mL) of organic phase, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give a white solid 2,5-hydroxycholest-5(4)-en-3-one (248-1) (767 mg, purity: 90%, yield: 69.38%).

[1004] 1 H NMR (400MHz, CDCl3) δ5.72 (s, 1H), 2.47-2.27 (m, 4H), 2.05-2.01 (m, 2H), 1.87-1.82 (m, 2H), 1.70 -1.24(m, 15H), 1.22(s, 6H), 1.18(s, 3H), 1.12-1.00(m, 5H), 0.93(d, J=6.5Hz, 3H), 0.71(s, 3H). 13 C NMR (101MHz, CDCl3) δ199.69, 171.71, 123.76, 71.09, 56.03, 55.87, 53.80, 44.39, 42.41, 39.62, 38.60, 36.3 8, 35.72, 35.69, 35.61, 33.99, 32.95, 32.04, 29.36, 29.23, 28.20, 24.17, 21.02, 20.77, 18.61, 17.39, 11.96.

[1005] In the second step, 25-hydroxycholest-5(4)-en-3-one (248-1) (600 mg, 1.50 mmol) was dissolved in methanol (25 mL) in a 50 mL round-bottom flask. Heptahydrate, cerium chloride (1.39 g, 3.75 mmol), and sodium borohydride (141.63 mg, 3.75 mmol) were added at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC (ethyl acetate / petroleum ether = 1 / 4). After the reaction was complete, the reaction mixture was added to ice water (50 mL), extracted with dichloromethane (30 mL × 3), washed with saturated brine (60 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 (eluent: ethyl acetate / petroleum ether = 20%), and the solvent was removed by concentration under vacuum to obtain a white solid cholester-5(4)-ene-3β,25-diol (248-2) (510 mg, purity: 80%, yield: 67.66%). 1 H NMR (400MHz, CDCl3) δ5.27 (d, J=1.5Hz, 1H), 4.17-4.13 (m, 1H), 2.23-2.15 (m, 2H), 2.04-1.96 (m, 4H), 1.86-1. 69 (m, 6H), 1.43-1.35 (m, 12H), 1.21 (s, 6H), 1.14-1.05 (m, 5H), 1.05 (s, 3H), 0.92 (d, J=6.6Hz, 3H), 0.68 (s, 3H)

[1006] In the third step, cholesterol-5(4)-en-3β,25-diol (248-2) (450 mg, 1.12 mmol) was dissolved in a 50 mL round-bottom flask containing 10 mL of dichloromethane at room temperature. Triethylamine (340 mg, 3.36 mmol), 4-dimethylaminopyridine (27.31 mg, 0.224 mmol), and acetic anhydride (171.14 mg, 1.68 mmol) were added at room temperature, followed by stirring at room temperature for 2 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, the mixture was quenched with water (50 mL), extracted with ethyl acetate (30 mL × 3), washed with saturated brine (50 mL) on the organic phase, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give a white solid acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,5,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (248-3) (440 mg, purity: 90%, yield: 79.68%).1 H NMR (400MHz, CDCl3) δ5.26-5.20 (m, 2H), 2.24-2.15 (m, 1H), 2.05 (s, 3H), 2.02-1.95 (1m, 2H), 1.88-1.68 (m, 4H), 1.62 -1.27 (m, 168H), 1.21 (s, 6H), 1.18-1.08 (m, 3H), 1.06 (s, 3H), 1.03-0.95 (m, 1H), 0.92 (d, J=6.5Hz, 3H), 0.68 (s, 3H).

[1007] In the fourth 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,4,5,7,8,9,9a,9b,10,11,11a-tetradecanoic-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (248-3) (250 mg, 0.562 mmol) was dissolved in a 50 mL round-bottom flask containing 5 mL of dichloromethane. Then, m-chloroperoxybenzoic acid (145.51 mg, 0.843 mmol) was added at room temperature, followed by stirring for 2 hours at room temperature. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 5:1). After the reaction was completed, the solution was quenched with saturated sodium sulfite (20 mL) and saturated sodium bicarbonate aqueous solution (20 mL), extracted with dichloromethane (20 mL × 3), washed with saturated brine (20 mL) of organic phase, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give a white solid (2S,4aR,4bS,6aR,7R,9aS,9bS)-7-((R)-6-hydroxy-6-methylheptane-2-yl)-4a,6a-dimethylhexadecylhydrocyclopentane[7,8]phenanthrene[1,10a-b]-2-acetate (248-4) (230 mg, purity: 90%, yield: 79.92%). 1H NMR (400MHz, CDCl3) δ5.14-5.10 (m, 0.57H), 4.92 (d, J=3.6Hz, 1H), 4.31-4.26 (m, 1H) , 3.16 (d, J=3.3Hz, 0.5H), 2.13 (s, 3H), 2.10 (s, 1.73H), 2.03-1.94 (m, 2H), 1.93-1.8 8(m, 1H), 1.86-1.71(m, 5H), 1.69-1.33(m, 32H), 1.21(s, 9.44H), 1.11(s, 3H), 1.04( s, 1.72H), 1.03-0.97 (m, 4H), 0.92 (d, J=6.4Hz, 4.74H), 0.68 (s, 1.74H), 0.65 (s, 3H).

[1008] In step six, (2S,4aR,4bS,6aR,7R,9aS,9bS)-7-[(2R)-6-hydroxy-6-methylhept-2-yl]-4a,6a-dimethyl-1a,2,3,4,4a,4b,5,6,6a,7,8,9,9a,9b,10,11-hexadecylhydrocyclopenta[1′,2′:1,2]phenanthro[8,8a-b]oxacyclopropane-2-ol (248-4) (40 mg, 0.087 mmol) was dissolved in acetone (2 mL) in a 50 mL round-bottom flask. A 10% aqueous solution of perchloric acid (0.087 mL, 0.087 mmol) was added dropwise to the reaction mixture, followed by stirring at room temperature for 16 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 1:1). After the reaction was complete, saturated sodium bicarbonate (10 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (5 mL × 3). The extracted organic solution was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. The organic phase was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1). Acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-5a,6-dihydroxy-1-[(2R)-6-hydroxy [6-methylhept-2-yl]-9a,11a-dimethylhexadecyl-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester isomer 1 (248-6) (Rf = 0.24) (15 mg, purity: 80%, yield: 28.87%) and isomer 2 (248-5) (Rf value 0.4) (20 mg, purity: 80%, yield: 38.50%).

[1009] 248-5 (Rf = 0.4): 1H NMR (400MHz, CDCl3) δ5.46-5.16 (m, 1H), 3.65 (d, J=3.0Hz, 1H), 2.30-2.18 (m, 1H), 2.08 (s, 3H), 2.02-1.96 (m, 2H), 1.88-1.77 (m, 3H), 1. 76-1.70 (m, 2H), 1.63-1.52 (m, 7H), 1.47-1.35 (m, 10H), 1.21 (s, 6H), 1.18 (s, 3H), 1.11-1.01 (m, 5H), 0.92 (d, J=6.4Hz, 3H), 0.66 (s, 3H).

[1010] 248-6 (Rf = 0.24): 1 H NMR (400MHz, CDCl3) δ4.92 (d, J=3.6Hz, 1H), 4.44-3.99 (m, 1H), 2.13 (s, 3H), 1.99 (d, J=12.3Hz, 1H), 1.92-1.80 (m, 2H), 1 .75-1.50 (m, 11H), 1.47-1.31 (m, 12H), 1.21 (s, 6H), 1.11 (s, 3H), 1.07-0.98 (m, 4H), 0.92 (d, J=6.5Hz, 3H), 0.65 (s, 3H).

[1011] In step seven, 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 isomer (248-5) (Rf = 0.4) (15 mg, 0.031 mmol) was dissolved in a 50 mL round-bottom flask containing tetrahydrofuran (0.5 mL) and methanol (0.5 mL). Lithium hydroxide monohydrate (6.57 mg, 0.157 mmol) and water (0.3 mL) were added at room temperature, followed by stirring at room temperature for 2 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 1:2). After the reaction was complete, the mixture was diluted with water (10 mL), extracted with ethyl acetate (5 mL × 3), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:2) to give a white solid cholesterol-3β,4,5,25-tetraol isomer 1 (248).

[1012] 248: 1H NMR (400MHz, CDCl3) δ4.14 (s, 1H), 3.54 (s, 1H), 2.24-2.16 (m, 1H), 2.05-1.93 (m, 2H), 1.80-1.58 (m, 10H ), 1.51-1.33(m, 13H), 1.21(s, 6H), 1.15(s, 3H), 1.11-0.99(m, 5H), 0.92(d, J=6.4Hz, 3H), 0.66(s, 3H). 13 C NMR (101MHz, CDCl3) δ78.14, 75.68, 71.16, 68.40, 56.28, 56.14, 46.97, 44.41, 42.77, 39.96, 38.40, 36.43, 35.76, 34.5 6, 31.51, 30.65, 29.71, 29.34, 29.20, 28.27, 25.84, 25.69, 24.10, 20.81, 20.53, 18.62, 15.59, 12.16.LCMS: [M+H-2H2O] + =401

[1013] Referring to Example 248, the 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 isomer (248-5) (Rf = 0.4) was replaced with 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 isomer (248-5) (Rf = 0.4). S,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 isomer (248-6) (Rf = 0.24) yields a white solid cholesterol-3β,4,5,25-tetraol isomer 2 (249).

[1014] 249: 1 H NMR (400MHz, CDCl3) δ4.19-4.12 (m, 1H), 3.54 (d, J=3.5Hz, 1H), 2.24-2.17 (m, 1H), 2.04-1.94 (m, 2H), 1.83-1.5 8(m, 10H), 1.49-1.33(m, 13H), 1.21(s, 6H), 1.15(s, 3H), 1.10-0.99(m, 5H), 0.92(d, J=6.5Hz, 3H), 0.66(s, 3H). 13C NMR (101MHz, CDCl3) δ78.14, 75.67, 71.15, 68.40, 56.28, 56.14, 46.97, 44.41, 42.77, 39.96, 38.40, 36.43, 35.76, 34.56, 31.5 1, 31.44, 30.65, 30.19, 29.71, 29.34, 29.21, 28.27, 25.84, 25.69, 24.10, 20.81, 20.53, 18.62, 15.59, 12.16.LCMS: [M+H-2H2O] + =401

[1015] Example 250

[1016] Preparation of compound 250 5-fluorocholest-3β,4,25-triol

[1017]

[1018]

[1019] In the first step, (2S,4aR,4bS,6aR,7R,9aS,9bS)-7-((R)-6-hydroxy-6-methylheptane-2-yl)-4a,6a-dimethylhexadecylhydrocyclopentane[7,8]phenanthrene[1,10a-b]-2-acetate (10 mg, 0.022 mmol) was dissolved in a 50 mL round-bottom flask containing triethylamine trihydrofluoride (0.301 mL, 1.845 mmol). The mixture was stirred at 100 °C for 16 hours, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 3:1). After the reaction was completed, the mixture was diluted with water (10 mL), extracted with ethyl acetate (5 mL × 3), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give a white solid acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-5a-fluoro-6-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (15 mg, purity: 80%, yield: 23.00%). 1H NMR (400MHz, CDCl3) δ5.09 (dd, J=10.1, 5.3Hz, 1H), 3.81 (d, J=4.0Hz, 1H), 2.34 (t, J=7.5Hz, 1H), 2.26-2.20 (m, 1H) , 2.09 (s, 4H), 2.05-1.98 (m, 4H), 1.82-1.46 (m, 22H), 1.21 (s, 6H), 1.15 (s, 4H), 0.92 (d, J=6.4Hz, 4H), 0.66 (s, 3H). 19 F NMR (376 MHz, CDCl3) δ-165.13 (s, 1F).

[1020] In the second step, acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-5a-fluoro-6-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexadecylhydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (250-1) (12 mg, 0.025 mmol) was dissolved in a 50 mL round-bottom flask containing 1 mL of tetrahydrofuran and 1 mL of methanol. Lithium hydroxide monohydrate (5 mg, 0.125 mmol) and water (0.5 mL) were added at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 1:1). After the reaction was completed, the solution was diluted with water (10 mL), extracted with ethyl acetate (5 mL × 3), washed with saturated brine (10 mL) on the organic phase, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give a white solid 5-fluorocholest-3β,4,25-triol (250) (3.5 mg, purity: 100%, yield: 31.96%). 1 H NMR(400 MHz, CDCl3) δ4.02-3.96 (m, 1H), 3.74-3.69 (m, 1H), 2.02-1.96 (m, 2H), 1.86-1.79 (m, 2H), 1.75-1.64 (m, 5 H), 1.46-1.31 (m, 16H), 1.21 (s, 6H), 1.12 (s, 3H), 1.11-0.98 (m, 5H), 0.92 (d, J=6.5Hz, 3H), 0.66 (s, 3H). 19 F NMR (376 MHz, CDCl3) δ-165.74 (s, 1F). 13C NMR (101MHz, CDCl3) δ75.60, 71.14, 69.65, 68.45, 57.66, 56.40, 56.10, 55 .98, 46.77, 44.41, 42.67, 39.85, 36.41, 35.75, 34.41, 30.86, 30.84, 30.0 8, 29.87, 29.70, 29.33, 29.21, 28.26, 27.21, 27.10, 26.44, 25.82, 25.79, 25.26, 24.09, 20.80, 20.36, 18.61, 15.31, 15.24, 12.10.LCMS: (M-H2O-F) + =401

[1021] Example 251

[1022] Preparation of compound 251 24-[(2-fluorophenyl)(hydroxy)methyl]-5α-cholan-3β-ol

[1023]

[1024]

[1025] 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 acid-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoate (151-6) (2 g, 4.50 mmol) in methanol (50 mL). Concentrated sulfuric acid (98%) (1 mL, 18.76 mmol) was added dropwise, followed by heating to reflux and stirring for 1 hour. After the reaction was complete, TLC (petroleum ether:ethyl acetate = 10:1) was used to monitor the reaction. Most of the solvent was then removed, and ethyl acetate (~100 mL) was added. The mixture was washed twice with saturated sodium bicarbonate solution (~50 mL x 10 mL). 2) Wash once with saturated brine (~50 mL), dry the organic phase with anhydrous sodium sulfate, filter, concentrate the filtrate to dryness, and purify the crude product by rapid 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-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecanoic acid methyl ester (269-1) (1.8 g, 4.02 mmol, yield 89.5%). 1H 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).

[1026] In the second step, compound (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 (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 (ester = 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 compound (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).

[1027] In the third 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-tetradecanoic acid methyl ester (269-2) (1.8 g, 3.58 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. After stirring at room temperature for 30 min, the mixture was monitored by TLC (petroleum ether:ethyl acetate = 10:1). The reaction was measured. After the reaction was completed, 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) to give 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, yield 90.0%). 1 H NMR (400MH...

Claims

1. A compound, or a pharmaceutically acceptable salt thereof, characterized in that, The compound has a structure represented by Formula XXI or XVI: wherein, R 3d is H; R 4a is H; R 4b is H or OH; or, R 4a and R 4b together with the carbon atom to which they are attached form R 8a is H; R 7a and R 7b are as defined in any one of the following groups: a) R 7a is F; R 7b is F; and b) R 7a and R 7b and the carbon atoms to which they are attached form R 14a is H; In the compound of formula XXI, R 22 is or R 21 ; In the compound of formula XXI, R 21 is -L 2 -R C ; In the compound of formula XXI, L 2 is -(CH2)3-, In the compound of formula XXI, R C is -C(R 21c )(R 21d )-OH; In the compound of formula XXI, R 21c is H, F, C 1-4 alkyl, fluoroC 1-4 alkyl, or C 3-6 cycloalkyl; R 21d is F, C 2-4 alkyl, fluoroC 1-4 alkyl, C 3-6 cycloalkyl, or phenyl, wherein the phenyl is independently unsubstituted or substituted with j R d groups; or, R 21c and R 21d together with the carbon atom that connects them form a C 3-6 cycloalkyl; in the compound represented by Formula XXI, j is 1, 2, 3, or 4; each R in the compound of formula XXI d independently F, Cl, C 1-4 alkyl, fluoroC 1-4 alkyl, C 1-4 alkoxy or fluoroC 1-4 alkoxy; In the compound of Formula XVI, R 3d is H or R 3c ; R 3c is -C(O)-CH2CH2COOH, -C(O)-CH2COOH or -C(O)-CH2OH; R 4a and R 4b are defined as in any one of the following groups: a) R 4a is H, R 4b is OH; b) R 4a is F, R 4b is F; c) R 4a is OH, R 4b is CF3; d) R 4a and R 4b and the carbon atoms to which they are attached collectively form e) R 4a and R 4b and the carbon atoms to which they are attached collectively form and f) R 4a is H, R 4b is H; In the compound of formula XVI, R 5 is H or R 5c ; R 5c is OH or F; In the compound of formula XVI, R 22 is R 21 ; In the compound of formula XVI, R 21 is or -L 2 -R C ; In the compound of formula XVI, L 2 is -(CH2)2- or -(CH2)3, In the compound of Formula XVI, each R C independently is -C(R 21c )(R 21d )-OH; In the compound of formula XVI, R 21c is H, F, C 1-4 alkyl, fluoroC 1-4 alkyl, or C 3-6 cycloalkyl; R 21d is F, C 2-4 alkyl, fluoroC 1-4 alkyl, C 3-6 cycloalkyl, or phenyl, wherein the phenyl is independently unsubstituted or substituted with j R d groups; or, R 21c and R 21d together with the carbon atom that connects them form a C 3-6 cycloalkyl; in the compound represented by Formula XVI, j is 1, 2, 3, or 4; each R in the compound of Formula XVI is independently F, Cl, C d independently F, Cl, C 1-4 alkyl, fluoroC 1-4 alkyl, C 1-4 alkoxy or fluoroC 1-4 alkoxy; the *-labeled carbon atom is in the R configuration, the S configuration, or a mixture of the two; the #-labeled carbon atom is a chiral carbon atom in the R configuration, the S configuration, or a mixture of the two; the &-labeled carbon atom is a chiral carbon atom in the R configuration, the S configuration, or a mixture of the two; the A-labeled carbon atom is a chiral carbon atom in the R configuration, the S configuration, or a mixture of the two; the B-labeled carbon atom is a chiral carbon atom in the R configuration, the S configuration, or a mixture of the two.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, In the compound of formula XVI, wherein R 3d is H or R 3c ; R 3c is -C(O)-CH2COOH or -C(O)-CH2OH; in Formula XVI, the *-labeled carbon atom is in the R configuration, the S configuration, or a mixture of the two; the #-labeled carbon atom is a chiral carbon atom in the R configuration, the S configuration, or a mixture of the two; the &-labeled carbon atom is a chiral carbon atom in the R configuration, the S configuration, or a mixture of the two.

3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein satisfies one or more of the following conditions: (1) in the compound of formula XXI, R C -C(R 21c )(R 21d )-OH is (2) in the compound of Formula XVI, R C -C(R 21c )(R 21d )-OH is 4. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein In the compound of formula XXI, R C -C(R 21c )(R 21d )-OH is 5. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein satisfies one or more of the following conditions: (1) in the compound of Formula XXI, R 21c is H, R 21d is C 2-4 alkyl, fluoroC 1-4 alkyl, C 3-6 cycloalkyl, or phenyl, wherein the phenyl is optionally substituted with one or two R d each R d is independently F or C 1-4 alkoxy; and (2) in the compound of Formula XVI, R 21c and R 21d are as defined in any one of the following groups: a) R 21c is H, R 21d is C 2-4 alkyl, fluoroC 1-4 alkyl, C 3-6 cycloalkyl or phenyl, wherein the phenyl is optionally substituted with 1 or 2 R d each R d is independently F or C 1-4 alkoxy; and b) R 21c is CH3or trifluoromethyl, R 21d is C 2-4 alkyl or trifluoromethyl.

6. The compound of claim 5, or a pharmaceutically acceptable salt thereof, wherein In the compound of formula XVI, -C(R 21c )(R 21d )-OH is 7. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein satisfies one or more of the following conditions: (1) in the compound of Formula XXI, R 21c is H, R 21d is phenyl, wherein the phenyl is optionally substituted with one or two R d , or R 21c and R 21d together with the carbon atom attaching them form a C 3-6 cycloalkyl; each R d is independently F or C 1-4 alkoxy; and (2) in the compound of Formula XVI, R 21c and R 21d are as defined in any one of the following groups: a) R 21c is H, R 21d is C 2-4 alkyl, fluoroC 1-4 alkyl, C 3-6 cycloalkyl or phenyl, wherein the phenyl is optionally substituted with 1 or 2 R d each R d is independently F, Cl, OH, fluoroC 1-4 alkyl, C 1-4 alkoxy or fluoroC 1-4 alkoxy; b) R 21c and R 21d and the carbon atoms attaching them together form a C 3-6 cycloalkyl; and c) R is CH3or trifluoromethyl, R 21c is CH3or trifluoromethyl, R 21d is phenyl or trifluoromethyl, wherein the phenyl is unsubstituted or substituted by j R d substituents.

8. The compound of claim 7, or a pharmaceutically acceptable salt thereof, wherein satisfies one or more of the following conditions: (1) in the compound of Formula XXI, R 21c is H, R 21d is phenyl, wherein the phenyl is optionally substituted with one or two R d ; or R 21c and R 21d together with the carbon atom that connects them form a cyclopropyl group; each R d is independently F or C 1-4 alkoxy; (2) in the compound of Formula XVI, R 21c and R 21d are defined as described in any one of the following groups: a) R 21c is H, R 21d is C 2-4 alkyl, fluoroC 1-4 alkyl, C 3-6 cycloalkyl or phenyl, wherein the phenyl is optionally substituted with 1 or 2 R d each R d is independently F, Cl, OH, fluoroC 1-4 alkyl, C 1-4 alkoxy or fluoroC 1-4 alkoxy; b) R 21c and R 21d and the carbon atoms attaching them form a cyclopropyl group; and c) R 21c is CH3or trifluoromethyl, R 21d is phenyl or trifluoromethyl, wherein the phenyl is unsubstituted or substituted by j R d substituents.

9. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein In the compound of formula XXI, -L 2 -R C is 10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein satisfies one or more of the following conditions: (1) in the compound of formula XXI, R 21 is and (2) in the compound of Formula XVI, R 21 is 11. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein satisfies one or more of the following conditions: (1) the compound represented by Formula XXI is a compound represented by Formula XXI-1: in the formula XXI-1, #, B, R 4a , R 4b , R 7a , R 7b and R 22 are as defined in any one of claims 1, 3 to 5, 7 to 10; (2) the compound represented by Formula XVI is a compound represented by Formula XVI-1: in which the definitions of #, &, R 3d , R 4a , R 4b , R 5 and R 22 are as given in any one of claims 1 to 3, 5 to 8, 10.

12. The compound of claim 11, or a pharmaceutically acceptable salt thereof, wherein, satisfies one or more of the following conditions: the compound represented by Formula XXI is a compound represented by Formula XXI-2, XXI-3, or XXI-4: In formulae XXI-2, XXI-3 or XXI-4, B, R 7a , R 7b and R 22 are as defined in any one of claims 1, 3 to 5, 7 to 10.

13. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein the compound represented by Formula XVI is a compound represented by Formula VII: wherein R 3d is H or R 3c ; R 3c is as defined in claim 1 or 2; R 22 R 21 ; R 21 as defined in any of claims 1, 3, 5-8, 10. in Formula VII, the *-labeled carbon atom is in the R configuration, the S configuration, or a mixture of the two.

14. A compound represented by any one of the following or a pharmaceutically acceptable salt thereof, characterized in that, 15. A pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, and at least one pharmaceutical excipient.

16. Use of a compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 15 for the manufacture of a medicament for inhibiting the SREBP pathway.

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