WO2025020190A1 - Steroid compounds, preparation method therefor and use thereof - Google Patents

Abstract

Disclosed in the present invention are steroid compounds, a preparation method therefor and the use thereof. Particularly disclosed are steroid compounds having a structure shown as formula (I), (II) or (III), etc., or pharmaceutically acceptable salts thereof. The compounds provided by the present invention have SREBP pathway inhibitory activity, and thus can be used for preventing and/or treating obesity, hyperlipidemia, fatty liver, diabetes, atherosclerosis, cardiovascular and cerebrovascular diseases, liver cancer, skin injury and other diseases.

Description

Steroidal compounds, preparation methods and applications thereof Technical Field

The present invention relates to a steroid compound, a preparation method and application thereof.

Background Art

With the change of lifestyle, including the intake of high-calorie food and high-sugar beverages, lack of exercise and physical activity, metabolic diseases represented by hyperlipidemia, obesity, type 2 diabetes and fatty liver have become increasingly serious health problems worldwide. Among them, fatty liver has become an important cause of chronic liver disease in Europe, America and wealthy areas of my country. The prevalence of simple liver lipid accumulation in ordinary adults is 10% to 30%, of which 10% to 20% is fatty hepatitis, and the incidence of cirrhosis and liver cancer in the latter reaches 25% within 10 years. However, as of now, the pathophysiological mechanism of fatty liver has not been fully elucidated, and there is still a lack of effective and specific therapeutic drugs in clinical practice. It is known that the accumulation of lipids such as cholesterol and triglycerides in the blood and liver is the main cause of hyperlipidemia, and hyperlipidemia is an important pathogenic factor causing atherosclerosis, cerebral stroke and fatty liver disease. Therefore, the development of new drugs targeting lipid metabolism regulation pathways with the orientation of lowering lipids is becoming an important direction for the development of new metabolic disease drugs.

It is known that the lipid synthesis pathway of mammalian cells is an important factor in regulating lipid metabolism balance. The key factor regulating the synthesis of cholesterol and fatty acids is a type of transcription factor protein, sterol response element binding protein SREBP (Sterol-Regulatory Element Binding Protein). The precursors of this type of protein are first synthesized on the endoplasmic reticulum (ER), and the precursors are transported to the Golgi apparatus through SREBP cleavage-activating protein (SCAP, SREBP cleavage-activating protein), and then cleaved by two proteases (Site-1protease (S1P) and Site-2protease (S2P)) to release its N-terminal active domain, enter the cell nucleus to play the role of a transcription factor, bind to the SREBP response element (SRE) in the promoter region of the target gene, and initiate the expression of downstream genes. The splicing and maturation of SREBP protein is strictly regulated by the level of intracellular sterols (such as cholesterol and 25-hydroxycholesterol). When cells accumulate sufficient cholesterol in the endoplasmic reticulum, cholesterol binds to SCAP and changes 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 apparatus and the subsequent activation of SREBP. Conversely, the active form of SREBP in the nucleus increases, promoting cellular lipid synthesis. In addition to cholesterol, 25-hydroxycholesterol (25-hydroxylcholesterol, 25-HC) is another potent endogenous inhibitor of the SREBP pathway. Unlike cholesterol binding to SCAP, 25-HC directly binds to Insig and induces the binding of SCAP and Insig.

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.

For hyperlipidemia, its pathogenesis is mainly due to increased lipid synthesis or abnormal lipid transport caused by factors such as diet or gene mutation, leading to excessive accumulation of lipids such as blood cholesterol and fatty acids. At present, statins and fibrates are the main lipid-regulating drugs in clinical practice. The mechanism of action of statins is to inhibit the cellular cholesterol synthesis pathway and promote the reverse transport of blood cholesterol. This shows that targeting the key factors of the cellular lipid synthesis pathway is an important means to effectively reduce lipid levels.

So far, there is no approved treatment for fatty liver disease, so it is very important to identify therapeutic targets and develop new effective therapies. It is known that the pathogenesis of fatty liver disease involves multiple risk factors, such as the accumulation of triglycerides in the form of lipid droplets, which may cause fatty degeneration, and abnormal increase in cholesterol and fatty acids in cells, which can cause endoplasmic reticulum stress and mitochondrial dysfunction, leading to cell death. Death, inflammation and fibrosis. Among them, free cholesterol accumulation has been reported as a key driver of the transition from simple steatosis to aggressive steatohepatitis. Secondly, a mouse model of fatty liver was established. A simple cholesterol-free high-fat diet could only induce steatosis even after long-term feeding, while adding 1-2% cholesterol to the diet was necessary to achieve inflammation and fibrosis. Therefore, lowering cholesterol may become a new therapeutic strategy for fatty liver disease. Studies have shown that abnormal activation of SREBPs was found in patients with fatty liver and mouse models of fatty liver; the deletion or knockout of mouse liver-specific Scap can eliminate the activation of all SREBPs, thereby preventing the occurrence of fatty liver and hyperlipidemia. In addition, recent studies have shown that abnormal activation of SREBPs induced by endoplasmic reticulum stress 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 for preventing and/or treating metabolic disorders including fatty liver.

Summary of the invention

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

The present invention provides a compound represented by formula I or a pharmaceutically acceptable salt thereof:

in,

R ^4a is H; R ^4b is halogen, CN, -CH ₂ -CN, -CH ₂ -OH, -CH(CH ₃ )-OH, -COOH, -CH ₂ -COOH, C ₁ -C ₆ haloalkyl, -CH ₂ -OAc or NH ₂ ; or, R ^4a and R ^4b together with the carbon atoms to which they are attached form

When there is a single bond between carbon atom No. 7 and carbon atom No. 8, R ^7a and R ^7b are independently H, halogen, CN, -CH ₂ -OH, -COOH, -CH ₂ -COOH, NH ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl or -CH ₂ -CN; or, R ^7a and R ^7b together with the carbon atoms to which they are connected form R ^8a is H;

When there is a double bond between carbon atoms 7 and 8, R ^7a does not exist, R ^7b is a halogen; R ^8a does not exist;

R ²¹ is

n1 and n3 are independently 2, 3, 4 or 5;

m1 and m3 are independently 0, 1, 2, 3, 4 or 5;

Ring A and Ring C are independently C ₆ -C ₁₀ aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or C ₃ -C ₆ cycloalkyl;

^RA and ^RC are independently halogen, _C1 - _C6 alkyl, C1- _{C6 haloalkyl} , _C1 - _C6 alkoxy or _C1 - _C6 haloalkoxy;

^R1 is or NR ^1a R ^1b ;

R ^1a and R ^1b are independently H or C ₁ -C ₆ alkyl;

A carbon atom with an "*" indicates that when it is a chiral carbon atom, it is in R configuration, S configuration, or a mixture of the two;

A carbon atom with a "#" indicates that when it is a chiral carbon atom, it is in R configuration, S configuration, or a mixture of the two;

A carbon atom with "&" indicates that when it is a chiral carbon atom, it is independently in R configuration, S configuration, or a mixture of the two;

The compound shown in formula I is not the following compound:

and its isomers.

In certain preferred embodiments of the present invention, certain groups in the compound or its pharmaceutically acceptable salt are defined as follows, and the unmentioned groups are the same as those described in any embodiment of the present invention (referred to as "in some embodiments"). In R ^4b , the halogen is independently F, Cl, Br or I; for example, Br.

In some embodiments, in R ^4b , the C ₁ -C ₆ haloalkyl is independently CHF ₂ , CH ₂ F or CF ₃ .

In some embodiments, in R ^7a and R ^7b , the halogen is independently F, Cl, Br or I; preferably F.

In some embodiments, in R ^7a and R ^7b , the C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

In some embodiments, in R ^7a and R ^7b , the C ₁ -C ₆ haloalkyl is independently CHF ₂ , CH ₂ F or CF ₃ , for example CHF ₂ .

In some embodiments, in Ring A and Ring C, the C ₆ -C ₁₀ aryl group is independently phenyl or naphthyl, preferably phenyl.

In some embodiments, in Ring A and Ring C, the “heteroatom selected from 1, 2 or 3 of N, O and S, and the number of heteroatoms is 1, 2 or 3 5-10 membered heteroaryl” is independently “the heteroatom selected from 1, 2 or 3 of N, O and S, 5-6-membered or 9-10-membered heteroaryl having 1 or 2 heteroatoms, for example pyridyl (e.g. ).

In some embodiments, in Ring A and Ring C, the C ₃ -C ₆ cycloalkyl group is independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

In some embodiments, in ^RA and ^RC , the halogen is independently F, Cl, Br or I; preferably F or Cl, such as F.

In some embodiments, in ^RA and ^RC , the C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

In some embodiments, in ^RA and ^RC , the C ₁ -C ₆ haloalkyl is independently CHF ₂ , CH ₂ F or CF ₃ .

In some embodiments, in ^RA and ^RC , the C ₁ -C ₆ alkoxy group is independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, preferably methoxy or ethoxy.

In some embodiments, in ^RA and ^RC , the C ₁ -C ₆ haloalkoxy group is independently -OCHF ₂ , -OCH ₂ F or -OCF ₃ ; preferably -OCF ₃ .

In some embodiments, in R ^1a and R ^1b , the C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as methyl.

In some embodiments, in Formula I, the carbon atom with "*" indicates that when it is a chiral carbon atom, it is in the S configuration.

In some embodiments, in Formula I, for

In some embodiments, R ^4a is H; R ^4b is halogen (eg, Br), CN, -CH ₂ -CN, -CH ₂ -OH, -CH(CH ₃ )-OH, -COOH, -CH ₂ -COOH, -CH ₂ -OAc, or NH ₂ ; or, R ^4a and R ^4b together with the carbon atoms to which they are attached together form For example, R ^4a is H; R ^4b is Br, CN, -CH ₂ -CN, -CH ₂ -OH, -CH(CH ₃ )-OH, -COOH, -CH ₂ - OAc or NH ₂ ; or, R ^4a and R ^4b together with the carbon atom to which they are attached together form

In some embodiments, when there is a single bond between carbon atom No. 7 and carbon atom No. 8, R ^7a and R ^7b are independently H or halogen (eg, F), preferably, R ^7a and R ^7b are both H or halogen (eg, F);

When there is a double bond between carbon atom No. 7 and carbon atom No. 8, R ^7a does not exist, R ^7b is halogen (such as F); R ^8a does not exist.

In some embodiments, R ²¹ is For example

In some embodiments, Ring A and Ring C are independently C ₆ -C ₁₀ aryl or "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S, and having 1, 2 or 3 heteroatoms".

In some embodiments, ^RA and ^RC are independently halogen, C ₁ -C ₆ alkoxy, or C ₁ -C ₆ haloalkoxy.

In some embodiments, n1 is 3.

In some embodiments, n3 is 3.

In some embodiments, m1 is 0, 1, 2 or 3.

In some embodiments, m3 is 0, 1, 2 or 3.

In some embodiments, for

In some embodiments, for

In some embodiments, for

In some embodiments, R ²¹ is

The present invention also provides a compound represented by formula II or a pharmaceutically acceptable salt thereof:

in,

R ^4c is H; R ^4d is H or OH;

R ^7c is H, R ^7d is independently CN, -CH ₂ -OH, -COOH, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, -CH ₂ -COOH, NH ₂ , -CH ₂ -CN or Cl; or, R ^7c and R ^7d together with the carbon atom to which they are attached form

R ²² is

n2 is 2, 3, 4 or 5;

m2 is 0, 1, 2, 3, 4 or 5;

Ring B is C ₆ -C ₁₀ aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or C ₃ -C ₆ cycloalkyl;

^RB is independently _{halogen, C1-C6 alkyl, C1-C6 haloalkyl , C1 - C6} alkoxy or _C1 - _C6 haloalkoxy;

A carbon atom with an "*" indicates that when it is a chiral carbon atom, it is in R configuration, S configuration, or a mixture of the two;

A carbon atom with a "#" indicates that when it is a chiral carbon atom, it is in R configuration, S configuration, or a mixture of the two;

A carbon atom with an "&" indicates that it is an R configuration, an S configuration, or a mixture of the two when it is a chiral carbon atom;

The compound shown in formula II is not the following compound:

and its isomers.

In some embodiments, in R ^7c and R ^7d , the C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

In some embodiments, in R ^7c and R ^7d , the C ₁ -C ₆ haloalkyl is independently CHF ₂ , CH ₂ F or CF ₃ , such as CHF ₂ .

In some embodiments, in ring B, the C ₆ -C ₁₀ aryl group is phenyl or naphthyl, preferably phenyl.

In some embodiments, in ring B, the “heteroatom selected from 1, 2 or 3 of N, O and S, 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms” is “heteroatom selected from 1, 2 or 3 of N, O and S, 5-6 membered or 9-10 membered heteroaryl group having 1 or 2 heteroatoms”, for example, pyridyl (for example ).

In some embodiments, in ring B, the C ₃ -C ₆ cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

In some embodiments, in ^RB , the halogen is independently F, Cl, Br or I; preferably F or Cl, such as F.

In some embodiments, in ^RB , the _C1 - _C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isopropyl, butyl, sec-butyl or tert-butyl.

In some embodiments, in ^RB , the C ₁ -C ₆ haloalkyl is independently CHF ₂ , CH ₂ F or CF ₃ .

In some embodiments, in ^RB , the C ₁ -C ₆ alkoxy group is independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, preferably methoxy or ethoxy.

In some embodiments, in ^RB , the C ₁ -C ₆ haloalkoxy group is independently -OCHF ₂ , -OCH ₂ F or -OCF ₃ ; preferably -OCF ₃ .

In some embodiments, in Formula II, the carbon atom with "*" indicates that when it is a chiral carbon atom, it is in the S configuration.

In some embodiments, in Formula II, the carbon atom with "#" indicates that when it is a chiral carbon atom, it is in the R configuration.

In some embodiments, in Formula II, for

In some embodiments, R ^7c is H, R ^7d is independently CN, -CH ₃ , -CH ₂ -OH, -COOH, -CHF ₂ , -CH ₂ -COOH, NH ₂ , -CH ₂ -CN or Cl; or, R ^7c and R ^7d together with the carbon atoms to which they are attached together form For example, R ^7c is H, R ^7d is independently CN, -CH ₂ -OH, -CH ₃ , NH ₂ , -CH ₂ -CN, -CF ₂ H, -COOH, or R ^7c and R ^7d together with the carbon atoms to which they are attached form

In some embodiments, ^R22 is

In some embodiments, ring B is C ₆ -C ₁₀ aryl or "heteroatoms selected from 1, 2 or 3 of N, O and S, wherein the heteroatoms are selected from "5-10 membered heteroaryl having 1, 2 or 3 sub-members".

In some embodiments, ^RB is independently halogen, _C1 - _C6 alkoxy, or _C1 - _C6 haloalkoxy.

In some embodiments, n2 is 3.

In some embodiments, m2 is 0, 1, 2 or 3.

In some embodiments, for

In some embodiments, ^R22 is

In some embodiments, in Formula II, for

The present invention also provides a compound represented by formula III or a pharmaceutically acceptable salt thereof:

in,

R ^3a is H or C ₁ -C ₆ alkyl;

R ^3b is -O-(CH ₂ )nR ^3-1 , NR ^3-2 R ^3-3 or OH; when R ^3b is OH, R ^3a is C ₁ -C ₆ alkyl;

n is 1, 2, 3 or 4; R ^3-1 is OH or -C(O)OR ^3-1a ; R ^3-1a is H or C ₁ -C ₆ alkyl;

R ^3-2 and R ^3-3 are independently H or -S(O) ₂ R ^3-2a ; R ^3-2a is C ₁ -C ₆ haloalkyl or C ₁ -C ₆ alkyl;

R ^4e is H; R ^4f is H, OH or -O-(CH ₂ )mOR ^4-1 ; or R ^4e and R ^4f together with the carbon atoms to which they are attached form m is 1 or 2; R ^4-1 is C ₁ -C ₆ alkyl;

A carbon atom with an "*" indicates that when it is a chiral carbon atom, it is in R configuration, S configuration, or a mixture of the two;

A carbon atom with a "#" indicates that when it is a chiral carbon atom, it is in R configuration, S configuration, or a mixture of the two;

The compound shown in formula III is not the following compound:

and its isomers.

In some embodiments, in R ^3a , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as methyl.

In some embodiments, in R ^3-1a , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example tert-butyl.

In some embodiments, in R ^3-2a , the C ₁ -C ₆ haloalkyl group is CHF ₂ , CH ₂ F or CF ₃ , such as CF ₃ .

In some embodiments, in R ^4-1 , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as ethyl.

In some embodiments, in Formula III, for

In some embodiments, n is 1, 2, or 3.

In some embodiments, R ^3-2 is H and R ^3-3 is -S(O) ₂ R ^3-2a .

In some embodiments, m is 1.

In some embodiments, R ^3a is H, R ^3b is Alternatively, R ^3a is methyl and R ^3b is OH.

In some embodiments, R ^4e is H; R ^4f is H, OH or Or R ^4e and R ^4f together with the carbon atoms to which they are attached form

In some embodiments, in Formula III, for

The present invention also provides any of the following compounds or pharmaceutically acceptable salts thereof:

The present invention also provides a pharmaceutical composition, comprising the compound described in any one of the present invention or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient.

The present invention also provides a use of a compound according to any one of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to any one of the present invention in the preparation of a medicament for preventing and/or treating a disease, wherein the disease is obesity, hyperlipidemia, fatty liver, diabetes, atherosclerosis, cardiovascular and cerebrovascular diseases, liver cancer or skin damage.

The present invention also provides a use of the compound according to any one of the present invention or a pharmaceutically acceptable salt thereof or the pharmaceutical composition as described above in the preparation of a drug for inhibiting the SREBP pathway.

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

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

Definition and Description

Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient.

In this article, the term "substitution" or "substituent" means that the hydrogen atom in the group is replaced by a specified group. When the substitution position is not specified, the substitution can be in any position, but only a stable or chemically feasible chemical is formed. Examples are as follows: The structure represents that the hydrogen atoms on ring A are replaced by m1 ^RAs . When multiple ^RAs are present, each ^RA is the same or different.

When any variable (eg, ^RA ) occurs more than one time in a compound's composition or structure, its definition at each occurrence is independent.

As used herein, the term "alkyl" refers to a saturated linear or branched monovalent hydrocarbon group. C ₁ -C ₆ alkyl refers to an alkyl group having 1 to 6 carbon atoms. In some embodiments, C ₁ -C ₆ alkyl may be C ₁ -C ₄ alkyl. C ₁ -C ₄ alkyl includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl.

In this article, the term "haloalkyl" refers to a group formed by replacing one or more hydrogen atoms in an alkyl group with a halogen, wherein the definition of alkyl is as described above. Examples of haloalkyl include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, etc.

In this document, the term "alkoxy" refers to -O-alkyl, wherein the definition of alkyl is as described above. C ₁ -C ₄ alkoxy refers to -O-(C ₁ -C ₄ alkyl), wherein the definition of C ₁ -C ₄ alkyl is as described above, that is, C ₁ -C ₄ alkoxy can specifically be methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy.

As used herein, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic (e.g., cyclohexyl, cyclopentyl, cyclohexyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, cyclopentyl, cyclobutyl, cyclopentyl ... etc. The C _3-6 cycloalkyl group may specifically be C ₃ , C ₄ , C ₅ , or C ₆ cycloalkyl group. In some embodiments, the cycloalkyl group is monocyclic. In some embodiments, the cycloalkyl group is polycyclic (eg, cyclic, spirocyclic, or bridged).

Herein, the term "C _6-10 aryl" refers to phenyl or naphthyl.

In this article, the term "heteroaryl" refers to an aromatic monocyclic or fused ring group formed by carbon atoms and at least one heteroatom, wherein the heteroatoms are independently selected from 1, 2 or 3 of N, O and S. The 5-10 membered heteroaryl can specifically be a 5, 6, 7, 8, 9 or 10 membered heteroaryl, such as a 5-6 membered heteroaryl or an 8-10 membered fused heteroaryl. The 5-6 membered heteroaryl is monocyclic, and specific 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 heteroaryls include but are not limited to benzopyrrole, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazole, benzisothiazole, benzopyrazole, benzimidazole, benzopyridine, benzopyrimidine, benzopyrazine, thiazolothiazole, pyridopyridine, pyridopyrazine, Pyridopyrimidine,

In this article, the chemical structure Indicates the connection location. Included in cyclic groups and not specified When the ring atoms are connected, Attachment to any ring atom is possible, but is permitted only if it results in a stable or chemically feasible compound.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a suitable non-toxic organic acid, inorganic acid, organic base or inorganic base and a compound, which retains the biological activity of the compound. The organic acid may be any organic acid that is conventional in the art and can form a salt. The inorganic acid may be any inorganic acid that is conventional in the art and can form a salt. The organic base may be any organic base that is conventional in the art and can form a salt. The inorganic base may be any inorganic base that is conventional in the art and can form a salt.

In chemical structures, solid wedge-shaped bonds are used and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed key Indicates the relative configuration of a stereocenter. The configuration is not specified, that is, if there are configurational isomers in the chemical structure, the bond Can be or include both Two configurations (e.g. The ratio of carbon to carbon double bonds is 1:1). When the specific configuration of a carbon-carbon double bond is not specified, it may be in E or Z configuration. Stereoisomers may 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 a chiral column.

As used herein, the term "subject" includes any animal, preferably a mammal, and more preferably a human.

As used herein, the term "effective amount" refers to a sufficient amount of a drug or pharmaceutical agent that is non-toxic but can achieve the desired effect. The determination of the effective amount varies from person to person, depending on the age and general condition of the recipient, and also on the specific active substance. The appropriate effective amount in each case can be determined by a person skilled in the art based on routine experiments.

Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

The reagents and raw materials used in the present invention are commercially available.

The positive progressive effect of the present invention is that the present invention provides a new class of compounds, which have inhibitory activity on the SREBP pathway and can be used to prevent and/or treat diseases such as obesity, hyperlipidemia, fatty liver, diabetes, atherosclerosis, cardiovascular and cerebrovascular diseases, liver cancer, skin damage, etc.

DETAILED DESCRIPTION

The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

Preparation of key intermediates

Examples I & II

Intermediate I (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-difluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanal I

Preparation of Intermediate II (5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-fluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-tetradecahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-8-yl]hexanal II

Step 1: Dissolve (22E, 24S)-stigmaster-6(5), 22(23)-diene-3β-ol I-1 (5.00 g, 12.11 mmol, 1.0 eq) in dichloromethane (50 mL), place the reaction system in an ice-water bath, cool to about 5°C, add acetic anhydride (3.4 mL, 36.35 mmol, 3.0 eq), 4-dimethylaminopyridine (300 mg, 2.42 mmol, 0.2 eq) and triethylamine (8 mL, 60.57 mmol, 5.0 eq) to the reaction system in sequence, and stir the reaction system at room temperature for 2 hours. After the reaction was completed as monitored by TLC (petroleum ether: ethyl acetate = 10:1), the reaction solution was quenched with methanol (20 mL), the reaction solution was washed once with saturated sodium bicarbonate (~50 mL) and water (~50 mL), dried over anhydrous sodium sulfate, and concentrated. When the concentration was almost dry, methanol (~20 mL) was added, and the mixture was stirred for 30 minutes under ice bath, filtered, and the filter cake was rinsed with a small amount of methanol. The filter cake was dried to obtain a white solid acetic acid-(1R,3aS,3bS, 7S,9aR,9bS,11aR)-1-[(2R,3E,5S)-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-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester I-2 (5.30 g, purity 90.0%, yield 86.58%). ¹ H NMR (400MHz, CDCl ₃ )δ5.37(d,J＝4.8Hz,1H),5.09(ddd,J＝56.1,15.2,8.6Hz,2H),4.61(ddd,J＝15.9,9.0,4.2Hz,1H),2.32(d,J＝7.3Hz,2H),2.03(s,3H),2.0 1–1.92(m,2H),1.87(dd,J＝8.9,6.6Hz,2H),1.73–1.40(m,12H),1.30–1.07(m,6H),1.02(t,J＝3.3Hz,6H),0.87–0.79(m,9H),0.70(s,3H).

Step 2: Weigh 10.0 g, 22 mmol, 1 eq of acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E,5S)-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-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester I-2 and dissolve it in tetrahydrofuran (200 mL) and water (20.0 mL). Add pyridine (4.5 mL) at room temperature. L, 55mmol, 2.5eq), N-methylmorpholine oxide (10.30g, 88mmol, 4eq), potassium osmate (0.81g, 2.2mmol, 0.1eq), stirred at room temperature overnight, TLC (petroleum ether: ethyl acetate = 3:1) monitoring, some raw materials remain, there is an intermediate (vicinal diol formation), then sodium periodate (18.80g, 88mmol, 4eq) was added to the reaction solution at 0°C, stirred at room temperature for 1 hour, TLC (petroleum ether: ethyl acetate = 3:1) detection, some raw materials remain, the intermediate is converted into a product. Then, 50 mL of water was added, and 50 mL of ethyl acetate was used for extraction, dried, concentrated, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate = 60:1) to obtain acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E,5S)-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-tetradecahydro-1H-cyclopenta[1, 2-i] phenanthrene-7-yl ester I-3 (3.3 g, purity 60%, yield 19.8%) white solid 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-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (1.9 g, purity 90.0%, yield 21.20%). ¹ H NMR (400MHz, CDCl ₃ )δ9.50(d,J＝3.3Hz,1H),5.31(d,J＝5.1Hz,1H),4.54(dd,J＝6.4,4.2Hz,1H),2.34–2.23(m,3H),1.96(s,3H),1.8 9(dt,J＝6.6,3.6Hz,2H),1.83–1.72(m,3H),1.66–1.09(m,14H),1.06(d,J＝6.8Hz,3H),0.96(s,3H),0.66(s,3H).

Step 3: (Methoxymethyl)triphenylphosphonium chloride (55.21 g, 161.052 mmol, 5.0 eq) was dissolved in anhydrous tetrahydrofuran (100 mL), the system was cooled to -10 °C in a dry ice ethyl acetate bath, sodium bis(trimethylsilyl)amide (80.526 mL, 1 mol/L, 2.5 eq) was added, and the dry ice ethyl acetate bath was stirred for 30 minutes, and 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-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester I-3 (12.0 g, 13.4 mmol, 1.0 eq) was dissolved in anhydrous tetrahydrofuran (50 mL) and added to the reaction solution. The system was returned to room temperature and stirred for 30 minutes. TLC (petroleum ether: ethyl acetate) was used to obtain the reaction mixture. 100 mL of saturated sodium bicarbonate solution was slowly added to the reaction system, extracted with ethyl acetate (100 mL × 3), the organic phase was collected and washed with water (100 mL × 2), washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated to obtain yellow solid 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-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester I-4 (8 g, purity 90.0%, yield 65.0%), and the crude product was directly used for the next step reaction.

Step 4: 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-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester I-4 (8 g, 12.5 mmol, 1.0 eq) was dissolved in tetrahydrofuran (100 mL), and dilute hydrochloric acid (5 mol/L, 50 mL) was slowly added, and the reaction mixture was stirred at room temperature for 30 minutes. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was complete, 80 mL of water was added to the reaction, and the reaction solution was extracted with ethyl acetate (50 mL × 3), washed with saturated brine (20 mL × 3), dried over anhydrous sulfuric acid, and the organic phase was collected and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 50: 1 to 30: 1 to 4: 1) and concentrated to obtain a white solid acetic acid-(1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-1-[(2R)-1-formylpropan-2-yl]-9a, 11a-dimethyl-2,3,3a, 3b, 4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester I-5 (6.3 g, purity 90%, yield 48.63%). ¹ H NMR (400 MHz, CDCl ₃ )δ9.76(dd,J＝3.3,1.3Hz,1H),5.37(d,J＝4.9Hz,1H),4.65–4.55(m,1H),2.46 (dd,J＝15.6,2.8Hz,1H),2.32(d,J＝7.0Hz,2H),2.22–2.14(m,1H),2.03(s,3H ),2.03–1.93(m,2H),1.89–1.78(m,3H),1.66–1.40(m,9H),1.17(dddd,J＝16. 5,14.2,10.9,7.1Hz,6H),1.03(dd,J＝7.5,3.5Hz,6H),0.73(d,J＝5.3Hz,3H).

Step 5: (The reactant acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-formylpropan-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester I-5 (10.00 g, 25.87 mmol, 1.0 eq) was dissolved in anhydrous tetrahydrofuran (150 mL) Add (triphenyl-λ5-methylphosphine) methyl acetate (51.89 g, 155.21 mmol, 6.0 eq) and stir the reaction system at 90 ° C for 18 h. The reaction raw materials were consumed by NMR monitoring. 100 mL of water was added to the reaction system, extracted with ethyl acetate (100 mL × 3), the organic phase was collected and washed with water (100 mL × 2), washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was collected and concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 60: 1 to 30:1) to obtain a white solid (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hex-2-enoic acid methyl ester I-6 (9.0 g, purity 90%, yield 71.1%). ¹ H NMR (400 MHz, CDCl ₃ )δ6.95(ddd,J＝15.4,8.7,6.4Hz,1H),6.26(d,J＝11.6Hz,0H),5.82(d,J＝15.5Hz,1 H),5.37(d,J＝4.9Hz,1H),4.60(tdd,J＝10.9,6.6,4.2Hz,1H),3.72(d,J＝10.2Hz,3H ),2.35–2.24(m,3H),2.03(s,3H),2.01–1.91(m,3H),1.90–1.78(m,3H),1.68–1.4 0(m,8H),1.31–1.07(m,5H),1.02(s,3H),0.95(d,J=6.7Hz,3H),0.72–0.68(m,3H).

Step 6: The reactant (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-tetradecahydro-1H-cyclopenta[1, 2-a] phenanthren-1-yl] hex-2-enoic acid methyl ester I-6 (10 g, 22.59 mmol, 1.0 eq) was dissolved in a mixed solvent of tetrahydrofuran (100 mL) and methanol (50 mL), nickel chloride (2.93 g, 22.59 mmol, 1.0 eq) was added, sodium borohydride (1.28 g, 33.89 mmol) was slowly added, and the reaction system was stirred at room temperature for 1 h. The reaction was monitored by NMR. The raw material was consumed. 100 mL of water was added to the reaction system, and ethyl acetate (100 mL × 3) was used for extraction. The organic phase was collected and washed with water (100 mL × 2), washed with saturated 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 30: 1) to obtain a white solid (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-tetradecahydro-1H-cyclopenta[1, 2-a] phenanthrene-1-yl] hexanoic acid methyl ester I-7 (9 g, purity 90%, yield 80.6%). ¹ H NMR (400 MHz, CDCl ₃ )δ5.37(d,J＝4.9Hz,1H),4.66–4.54(m,1H),3.67(s,3H),2.29(ddd,J＝15.9,11. 9,6.6Hz,4H),2.03(s,3H),2.02–1.93(m,2H),1.82(ddd,J＝13.0,10.8,8.3Hz,3H ),1.62–1.38(m,11H),1.19(dddd,J＝32.6,25.0,13.7,6.9Hz,9H),1.01(d,J＝5. 8Hz, 3H), 0.93 (d, J＝6.6Hz, 3H), 0.87 (tdd, J＝10.2, 4.8, 2.0Hz, 4H), 0.67 (s, 3H).

Step 7: In a 250 mL round-bottom flask at room temperature, (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-7 (2.5 g, 5.62 mmol) was dissolved in chloroform (80 mL), N-methylmorpholine (1.71 g, 16.87 mmol) and selenium dioxide (1.56 g, 14.06 mmol) were added at room temperature, and then stirred at 70°C for 18 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was completed, the reaction mixture was quenched with water (100 mL), extracted with dichloromethane (80 mL×3), the organic phase was washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1) to obtain a white solid (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-8 (1.4 g, purity: 90%, yield: 48.65%). ¹ H NMR (400MHz, CDCl ₃ )δ＝5.75–5.65(m,1H),4.80–4.65(m,1H),4.25(d,J＝2.6,1H),3.67(s,3H),2.33–2.22(m,2H),2.10(s,3H) ,2.07–1.98(m,2H),1.92–1.36(m,17H),1.22(s,3H),1.19–1.06(m,5H),0.93(d,J=6.6,3H),0.68(s,3H).

Step 8: The reactant (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-8 (5 g, 11.5 mmol, 1.0 eq) was dissolved in methanol (150 mL), potassium carbonate (6.37 g, 46 mmol, 4.0 eq) was added, and the reaction system was stirred at room temperature for 30 min. TLC (petroleum ether: ethyl acetate = 5:1) was used to monitor the reaction. When the raw material was consumed, the reaction was stopped. Water (100 mL) was added to the reaction system, and the aqueous layer was extracted with ethyl acetate (3×50 mL). The ethyl acetate layers were combined and washed with saturated brine (3×50 mL). The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered and concentrated to give crude (5R)-5-[(1R, 3aS, 3bS, 6R, 7S, 9aR, 9bS, 11aR)-6,7-dihydroxy-9a, 11a-dimethyl-2,3,3a, 3b, 4,6,7,8,9,9a,9b, 10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-9, which was used directly in the next step.

Step 9: The reactant (5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-dihydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-9 (5 g, 11.9 mmol, 1 eq) was dissolved in acetone (150 mL), p-toluenesulfonic acid (1.59 g, 8.4 mmol, 0.7 eq) and 4A molecular sieves were added, and the reaction system was stirred at room temperature for 1 h. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 10:1). After the raw material was consumed, the reaction was stopped by quenching with saturated sodium sulfite. 100 mL of water was added to the reaction system, and ethyl acetate (50 mL × 3) was used for extraction. The organic phase was collected and dried over anhydrous sodium sulfate. The organic phase was vacuum dried to obtain a crude product. The crude product was dissolved in ethyl acetate and purified by column chromatography (petroleum ether: ethyl acetate = 95:5) to obtain a white solid. (5R)-5-[(3aR,3R,5aS,9aS,9bS)-7-[(4R)-2,2-dimethyl-1,3-dioxolan-4-yl]-3a,6,6-trimethyl-2,3,3a,4,5,5a,6,9,9a,9b-decahydro-1H-cyclopenta[1,2-a]naphthalen-3-yl]hexanoic acid methyl ester I-10 (3.8 g, purity 90%, yield 62%). 1H NMR (400MHz, CDCl3) δ5.80(d,J＝2.9Hz,1H),4.41(d,J＝5.8Hz,1H),4.10(dd ,J＝13.5,6.5Hz,1H),3.67(s,3H),2.35–2.20(m,2H),2.14–1.98(m,2H),1. 87–1.56(m,10H),1.53(s,3H),1.45–1.37(m,3H),1.35(s,3H),1.28–1.21( m,1H),1.16(s,3H),1.14–0.97(m,6H),0.93(d,J=6.5Hz,3H),0.69(s,3H).

Step 10: Compound (5R)-5-[(3aR,3R,5aS,9aS,9bS)-7-[(4R)-2,2-dimethyl-1,3-dioxolan-4-yl]-3a,6,6-trimethyl-2,3,3a,4,5,5a,6,9,9a,9b-decahydro-1H-cyclopenta[1,2-a]naphthalen-3-yl]hexanoic acid methyl ester I-10 (3.8 g, 8.2 mmol, 1.0 eq) was dissolved in acetone (50 mL), N-hydroxyphthalimide (0.54 g, 3.3 mmol, 0.8 eq), tert-butyl hydroperoxide (12 mL, 66 mmol, 8.0 eq) and anhydrous cobalt (II) acetate (0.29 g, 1.6 mmol, 0.2 eq) were added, and the resulting mixture was stirred under N ₂ , stirred at 25°C for 18 hours. TLC (petroleum ether: ethyl acetate = 10:1) monitored the reaction. The raw material was basically consumed, quenched with saturated sodium sulfite, water (10 mL) was added to the reaction system, and the aqueous layer was extracted with ethyl acetate (3×15 mL). The organic layers were combined and washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 87: 13) to give a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-11-oxyylidene-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester I-11 (2.5 g, purity 95%. Yield 60%). ¹ H NMR (400MHz, CDCl ₃ )δ5.85(s,1H),4.45(d,J＝6.4Hz,1H),4.26(dd,J＝11.3,5.5Hz,1H),3.60( s,3H),2.34–2.25(m,2H),1.86–1.70(m,2H),1.58(dddd,J＝16.6,11.8,6.3 ,2.3Hz,5H),1.50(s,3H),1.40(ddd,J＝16.5,9.3,3.9Hz,4H),1.30(s,3H) ,1.26(s,4H),1.12–0.97(m,4H),0.87(d,J＝6.6Hz,3H),0.66–0.59(m,3H).

Step 11: Compound (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-tetradecahydro-1H-cyclopenta[1, 2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-11 (2.4 g, 5.1 mmol, 1.0 eq) was dissolved in methanol (100 mL) and ethyl acetate (50 mL), and then palladium carbon (1.2 g, 11 mmol, 2.2 eq) was added and the reaction system was replaced with a hydrogen atmosphere, and the reaction system was stirred at room temperature for 1 h. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 5: 1), and the raw material was completely consumed. The reaction system was filtered, and the organic phase was collected and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 90:10) to give a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-2,2,5a,7a-tetramethyl-11-oxyylidene-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester I-12 (1.7 g, purity 95%, yield 67%). 1H NMR(400MHz, CDCl3)δ4.00–3.87(m,2H),3.60(s,3H),2.75–2.66(m,1H),2.3 6(t,J＝11.3Hz,1H),2.25–2.09(m,4H),1.85(dddd,J＝28.9,13.2,6.5,3.1Hz,3H) ,1.62(dddd,J＝20.4,14.3,6.7,3.3Hz,4H),1.46(s,3H),1.43–1.26(m,5H),1.23 (s,6H),0.98(ddt,J＝14.4,11.7,7.4Hz,5H),0.86(d,J＝6.6Hz,3H),0.59(s,3H).

Step 12: Compound (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-tetramethyl-11-oxylidene-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolane -8-yl]hexanoic acid methyl ester I-12 (1.7 g, 3.6 mmol) was dissolved in diethylaminosulfur trifluoride (10 mL), and the resulting mixture was stirred in the reaction solution at 80 ° C for 1 h. TLC plate (petroleum ether: ethyl acetate = 10: 1) monitored the reaction to be complete. The reaction solution was cooled to room temperature, dichloromethane (50 mL) was added to dilute the reaction system, and ice water was carefully added dropwise to quench the reaction. The organic phase was collected by stratification, the aqueous layer was extracted with dichloromethane (3×30 mL), and the combined organic phases were washed with saturated brine (40 mL), filtered and concentrated to obtain a crude product. The crude product was added to a silica gel column (petroleum ether: ethyl acetate = 93:7) to obtain a colorless transparent solid (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-difluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanoic acid A mixture of methyl ester I-13 and (5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-fluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-tetradecahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester II-1 (1.4 g, I-13 purity 95%, yield calculated based on I-13: 74%).

Compound I-13: ¹ H NMR (400MHz, CDCl ₃ )δ4.02(d,J＝15.0Hz,2H),3.67(s,2H),2.33–2.23(m,2H),2.18–1.58(m,6H),1.51(s,2H),1.46–1.3 4(m,2H),1.30(s,2H),1.13(dd,J＝17.2,7.0Hz,1H),1.07(s,1H),0.93(d,J＝6.3Hz,2H),0.68(s ,3H).

In the thirteenth step, the compound (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-difluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester I-13 and (5R)-5-[(3aS,5aR,5bR,7aR,8R,1 A mixture of 1,2-dioxol-2,4-dihydro-1,4-dihydro-2 ...2,4-dihydro-1,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-1,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-1,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-1,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-2,4-dihydro-2, The reaction mixture was quenched with sodium sulfate decahydrate, filtered and concentrated to give a white solid (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-difluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexan-1-ol I-14 and (5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-fluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-tetradecahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester II-12 crude product (0.9 g, purity 96%, yield: 92%), directly used for the next step.

Step 14: Compound (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-difluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexan-1-ol I-14 and (5R)-5-[(3aS,5aR,5bR,7aR A mixture of 1,8R,10aR,12aR,12bR)-11-fluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-tetradecahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester II-2 (1.3 g, 2.7 mmol, 1.0 eq) was dissolved in dichloromethane (50 mL), and Dess-Martin reagent (2.3 g, 5.5 mmol, 2 eq) was added, and the resulting mixture was stirred in the reaction solution under N ₂ at 25° C. for 1 h. TLC (petroleum ether:ethyl acetate=10:1) monitored the reaction to be complete. Saturated sodium bicarbonate (10 mL) was added to the reaction system, and the organic phase was extracted with saturated sodium sulfite (3×30 mL). The organic phase was collected and concentrated. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 92: 8) gave a white solid (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-difluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolane- A mixture of (5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-fluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-tetradecahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-8-yl]hexanal II (0.95 g, purity of I 90%, yield based on I 66%).

Compound I: ¹ H NMR (400 MHz, CDCl ₃ ) δ9.70 (t, J=1.7 Hz, 1H), 3.98–3.91 (m, 2H), 2.39–2.25 (m, 2H), 1.96–1.86 (m, 1H), 1.81–1.71 (m, 2H), 1.64 (ddd, J=10.7, 10.3, 4.7 Hz, 1H), 1.57–1.52 (m, 1H), 1.4 4(s,2H),1.38–1.28(m,2H),1.23(s,1H),1.18(d,J＝9.1Hz,1H),1.08–1.02(m,1H ),1.00(s,1H),0.87(d,J＝6.5Hz,2H),0.83–0.77(m,1H),0.60(d,J＝12.0Hz,2H).

Example III

Preparation of Intermediate III (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanal (III)

Step 1: Dissolve compound (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester I-10 (1 g, 2.18 mmol, 1.0 eq) in tetrahydrofuran (50 mL), replace with nitrogen, add lithium aluminum tetrahydride (0.12 g, 3.270 mmol, 1.5 eq), and stir at 25° C. for 1 h. Monitor the reaction progress with a TLC plate (petroleum ether:ethyl acetate=5:1), and the raw material is completely consumed. The solution was quenched with sodium sulfate decahydrate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 80:20) to give a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexan-1-ol III-1 (1 g, purity: 90%, yield: 96%). ¹ H NMR (400 MHz, CDCl ³ ) δ 5.80 (d, J = 2.6 Hz, 1H), 4.41 (d, J = 2.6 Hz, 1H), 4.87 (d, J = 3.9 Hz, 2H). ＝5.8Hz,1H),4.09(m,1H),3.64(t,J＝6.5Hz,2H),2.08(m,2H),1.84(tt,J＝18.5,7.8Hz,2H),1.67(m,8H),1.41(td,J ＝12.7,6.2Hz,4H),1.35(s,3H),1.24(m,2H),1.17(s,4H),1.08(m,5H),0.93(d,J＝6.5Hz,4H),0.69(d,J＝4.6Hz,3H).

Step 2: Compound (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexan-1-ol III-1 (1 g, 90% purity, 2.09 mmol) was dissolved in dichloromethane (50 mL), Dess-Martin periodinane (1.18 g, 2.79 mmol) was added, and stirred at 25° C. for 1 h. TLC (petroleum ether:ethyl acetate=5:1) was used to monitor the reaction progress and the consumption of the starting materials. The mixture was quenched with saturated sodium sulfite, water (10 mL) was added to the reaction system, and the aqueous layer was extracted with dichloromethane (3×50 mL). The organic phases were combined and washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=90:10) to give a white solid (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanal III (750 mg, 1.58 mmol, purity: 90%, yield: 68%). ¹ H NMR (400MHz, CDCL3) δ9.76 (s, 1H), 5.80 (d, J = 2.7Hz, 1H), 4.41 (d, J = 5.8Hz, 1H ),4.10(dd,J＝13.6,6.3Hz,1H),2.39(m,2H),2.12(m,1H),2.01(m,1H),1.83(m ,1H),1.66(m,10H),1.53(s,4H),1.42(dd,J＝12.5,4.2Hz,3H),1.35(s,3H),1 .17(s,3H),1.10(m,4H),0.92(dd,J＝18.2,5.8Hz,4H),0.71(d,J＝12.1Hz,3H).

Example 9 & 10

Compound 9 3β-[(2-hydroxyethyl)oxy]-5α-cholest-25-ol and

Compound 10 {[(1R, 3aS, 3bR, 5aS, 7S, 9aS, 9bS, 11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetate-2-methylpropan-2-yl ester

Step 1: Dissolve (1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-methoxy-6-oxylidenehexan-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl acetate I-7 (80 mg, 0.180 mmol) in tetrahydrofuran (2 mL) and methanol (1 mL), add Pd(OH) ₂ (20 mg, 0.142 mmol), replace the atmosphere with hydrogen three times, stir at 40 °C for 48 hours, and monitor by HNMR. The reaction solution was filtered and the filtrate was concentrated to obtain acetic acid-(1R,3aS,3bR,7S,9aS,9bS,11aR)-1-[(2R)-6-methoxy-6-oxyylidenehexan-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 9-1 (80 mg, 0.170 mmol, 94.57%) as an ^off -white solid. HNMR(399MHz,cdcl3)δ4.74–4.58(m,1H),3.64(s,3H),2.32–2.17(m,2H),2.00(s,3H),1. 92(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).

Step 2: Dissolve methylacetic acid-(1R,3aS,3bR,7S,9aS,9bS,11aR)-1-[(2R)-6-methoxy-6-oxyylidenehexan-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 9-1 (80 mg, 0.179 mmol) in tetrahydrofuran (3 mL), replace with nitrogen three times, cool to -78 ° C, add methyl lithium (0.560 mL, 0.896 mmol), warm to room temperature and stir for 3 hours. Monitor 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 with brine once, dried over anhydrous sodium sulfate, and concentrated to obtain 80 mg of crude product. Cholesterol-3β,25-diol 9-2 (20 mg, 0.040 mmol, 22.08%) was obtained by column chromatography (DCM:MeOH=300:1-100:1). ¹ H NMR (399MHz, CDCl ₃ )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,3 H),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 CNMR (101MHz, CDCl ₃ )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.

Step 3: Dissolve 5α-cholest-3β,25-diol (10 mg, 0.025 mmol) in toluene (3 mL), add potassium tert-butoxide (14.03 mg, 0.125mmol), stirred at room temperature for 1 hour. Add tert-butyl bromoacetate (24.38mg, 0.125mmol), stir at room temperature for 2 hours. Monitor by TLC (petroleum ether: ethyl acetate = 3:1). The reaction solution was directly concentrated to obtain 40mg of crude product, which was purified by column chromatography (ethyl acetate: petroleum ether = 30%) to obtain {[(1R, 3aS, 3bR, 5aS, 7S, 9aS, 9bS, 11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetate-2-methylpropan-2-yl ester 10 (3mg, 0.005mmol, 22.23%) as a white solid.

Compound 10: ¹ HNMR (399MHz, Chloroform-d) δ3.98 (s, 2H), 3.29 (dt, J = 11.2, 6.2 Hz, 1H), 1.90 (dd, J = 30.3, 12.5 Hz, 2H), 1.79–1.67 (m, 2H), 1.62 (d, J = 13.0 Hz, 4H), 1.51 (d, J = 7.2 Hz, 5H), 1.45(s,9H),1.42(d,J＝2.1Hz,5H),1.36–1.29(m,6H),1.19(s,6H),1.04(dt,J＝20.1 ,9.4Hz,6H),0.89(d,J＝6.5Hz,3H),0.84(d,J＝14.0Hz,2H),0.77(s,3H),0.62(s,3H).

Step 4: Dissolve {[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetate-2-methylpropan-2-yl ester 10 (20 mg, 0.039 mmol) in tetrahydrofuran (5 mL), add lithium aluminum tetrahydride (4.39 mg, 0.116 mmol), and stir at room temperature for 1 hour. Monitor by TLC (petroleum ether: ethyl acetate = 3:1). The reaction solution is directly concentrated to obtain 10 mg of crude product, which is purified by column chromatography (ethyl acetate: petroleum ether = 40%) to obtain 3β-[(2-hydroxyethyl)oxy]-5α-cholest-25-ol 9 (10 mg, 0.021 mmol, 54.92%) as a white solid.

Compound 9: ¹ H NMR (399 MHz, Chloroform-d) δ3.69 (t, J = 4.5 Hz, 2H), 3.55 (td, J = 4.5, 2.2 Hz, 2H), 3.29–3.20 (m, 1H), 2.10–1.91 (m, 2H), 1.87–1.75 (m, 2H), 1.71 (dd, J = 13.4, 3.9 Hz, 1H), 1.62 (d, J = 3.3 Hz, 2H), 1.52 (s, 1H), 1.49–1.39 (m, 3H), 1.3 7(s,2H),1.31(d,J＝6.6Hz,2H),1.27(d,J＝4.6Hz,2H),1.23(d,J＝7.1Hz,3H),1.19(s,6H),1.13–1.02(m,4H),1.0 1–0.91(m,3H),0.89(d,J＝6.6Hz,3H),0.88–0.79(m,2H),0.78(s,3H),0.63(s,3H),0.58(dd,J＝10.7,4.0Hz,1H). ¹³ C NMR (101MHz, CDCl ₃ )δ79.02,71.10,68.82,62.11,56.46,56.18,54.35,44.78,44.39,42.59,40.00,36.91,36.40,35.73,3 5.46,34.79,32.07,29.32,29.17,28.81,28.24,24.18,21.20,20.74,18.61,12.26,12.0.LCMS:[M+Na] ⁺ =458.39.

Example 12

Compound 12 (5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-7-amino-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester

Step 1: Dissolve (1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-methoxy-6-oxyylidenehexan-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl acetate I-7 (1 g, 2.249 mmol) in tetrahydrofuran (5 mL) and methanol (5 mL). Add potassium carbonate (3.11 g, 22.489 mmol). Stir at 40°C for 5 hours and monitor by TLC (petroleum ether: ethyl acetate = 3:1). The reaction solution is concentrated to obtain 5 g of crude product. Column chromatography purification (ethyl acetate/petroleum ether = 40%) gave (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester ^12-1 (0.8 g, 1.888 mmol, 83.94%) as a white solid. NMR(399MHz,Chloroform-d)δ5.33(d,J＝5.1Hz,1H),3.65(s,3H),3.54–3.47(m,1H),2.33–2.17(m,5H), 2.00–1.92(m,2H),1.90–1.74(m,4H),1.67(d,J＝7.9Hz,1H),1.53–1.46(m,5H),1.43(dd,J＝11.2,4.6Hz ,3H),1.40–1.32(m,2H),1.27–1.12(m,4H),1.07(dd,J＝12.8,7.1Hz,4H),1.03(d,J＝4.8Hz,1H),0.99(s ,3H),0.95(d,J＝5.9Hz,1H),0.92(d,J＝6.6Hz,3H),0.85(dd,J＝24.1,6.0Hz,1H),0.66(d,J＝2.2Hz,3H).

Step 2: In a 50 mL round-bottom flask at room temperature, (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 12-1 (250 mg, 0.62 mmol) was dissolved in ethyl acetate (10 mL), acetic acid (0.2 mL) and platinum dioxide (125 mg) were added at room temperature, and the mixture was stirred in a hydrogen atmosphere at room temperature for 16 hours. The reaction was detected by TLC (ethyl acetate/petroleum ether = 5:1), and a new compound was generated. After the reaction, the reaction solution was diluted with ethyl acetate (30 mL), filtered through diatomaceous earth, the organic phase was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain a white solid (5R)-5-[(1R, 3aS, 3bR, 5aS, 7S, 9aS, 9bS, 11aR)-7-hydroxy-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 12-2 (240 mg, purity: 90%, yield: 85.97%). ¹ H NMR (400MHz, CDCl3) δ3.66(s,3H),3.62–3.54(m,1H),2.32–2.21(m,2H),1.84–1.75(m,3H),1.74–1.61(m,5H) ),1.58–1.46(m,5H),1.42–1.22(m,11H),1.14–1.04(m,5H),0.91(d,J＝6.5Hz,3H),0.80(s,3H),0.64(s,3H).

Step 3: In a 50 mL round-bottom flask at room temperature, (5R)-5-[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-7-hydroxy-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 12-2 (230 mg, 0.57 mmol) was dissolved in 1,2-dichloroethane (10 mL). Dess-Martin periodinane (362 mg, 0.85 mmol) was added at room temperature, and the mixture was stirred at 45° C. for 3 hours. The reaction was monitored by TLC (ethyl acetate/petroleum ether=10:1). After the reaction, the reaction solution was diluted with dichloromethane (10 mL), and saturated aqueous sodium sulfite solution (20 mL) and saturated aqueous sodium bicarbonate solution (20 mL) were added to quench the reaction. The organic phase was extracted with dichloromethane (20 mL×3), washed with saturated brine (30 mL), and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1) to obtain a white solid (5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-9a,11a-dimethyl-7-oxyylidenehexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 12-3 (220 mg, purity: 90%, yield: 86.52%). ¹ H NMR (400MHz, CDCl3) δ = 3.67 (s, 3H), 2.42–2.22 (m, 5H), 2.10–1.96 (m, 3H), 1.85–1.77 (m, 1H), 1.73–1.65 (m, 2H),1.60–1.48(m,6H),1.43–1.30(m,7H),1.27–1.04(m,6H),1.00(s,3H),0.92(d,J=6.5,3H),0.67(s,3H). ¹³ C NMR (101MHz, CDCl ₃ )δ=212.15,174.33,56.27,55.93,53.78,51.43,46.70,44.73,42.61,39.87,38.56,38.19,35.65, 35.51,35.40,35.39,34.52,31.70,29.70,28.96,28.16,24.20,21.55,21.43,18.55,12.04,11.46.

Step 4: In a 50 mL round-bottom flask at room temperature, (5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-9a,11a-dimethyl-7-oxoylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 12-3 (190 mg, 0.47 mmol) was dissolved in tetrahydrofuran (3 mL) and methanol (6 mL), and ammonium acetate (363 mg, 4.70 mmol) and a catalytic amount of acetic acid (0.20 mL) were added at room temperature and stirred at room temperature for 2 hours. Then sodium cyanoborohydride (297 mg, 4.70 mmol) was added to the reaction solution and stirred at room temperature for 14 hours. The reaction was monitored by TLC (dichloromethane/methanol = 10:1). After the reaction, the reaction solution was diluted with dichloromethane (20 mL), saturated sodium bicarbonate aqueous solution (30 mL) was added to quench the reaction, and the organic phase was extracted with dichloromethane (20 mL×3). The organic phase was washed with saturated brine (30 mL) and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to obtain a white solid (5R)-5-[(1R, 3aS, 3bR, 5aS, 9aS, 9bS, 11aR)-7-amino-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 12-4 (160 mg, purity: 90%, yield: 75.59%). ¹ H NMR (400MHz, DMSO) δ7.68 (s, 2H), 3.57 (s, 3H), 2.94 (dd, J = 25.9, 13.9Hz, 1H), 2.32–2.16 (m, 2H),1.98–1.88(m,1H),1.80–0.95(m,27H),0.88(d,J＝6.4Hz,3H),0.76(s,3H),0.62(s,3H).

Step 5: In a 50 mL round-bottom flask at room temperature, (5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-7-amino-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 12-4 (160 mg, 0.396 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL). The reaction was cooled to 0°C under nitrogen protection, and 3 mol/L methyl magnesium bromide tetrahydrofuran solution (1.321 mL, 3. 963mmol), then stirred at room temperature for 2 hours, and the reaction was monitored by TLC (dichloromethane: methanol = 8: 1). After the reaction, the system was cooled to 0°C and quenched with saturated aqueous ammonium chloride solution (20mL), extracted with ethyl acetate (15mL×3), and the organic phase was washed with saturated brine (30mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (dichloromethane: methanol = 8: 1) to obtain 3-amino-5α-cholest-25-ol (130mg, purity: 80%, yield: 64.99%) as a white solid. In a 50 mL round-bottom flask at room temperature, (5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-7-amino-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester (160 mg, 0.396 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL). The reaction was cooled to 0°C under nitrogen protection, and 3 mol/L methyl magnesium bromide tetrahydrofuran solution (1 .321mL, 3.963mmol), then stirred at room temperature for 2 hours, monitored by TLC (dichloromethane: methanol = 8: 1), after the reaction, the system was cooled to 0 ° C, quenched with saturated aqueous ammonium chloride solution (20mL), extracted with ethyl acetate (15mL × 3), the organic phase was washed with saturated brine (30mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (dichloromethane: methanol = 8: 1) to obtain a white solid 3-amino-5α-cholest-25- Alcohol 12 (130 mg, purity: 80%, yield: 64.99%). (ESI-MS (M+H)+: 404.4. ¹ H NMR (400 MHz, DMSO) δ7.61 (s, 2H), 4.03 (s, 1H), 2.93 (m, 1H), 1.93 (d, J = 12.0 Hz, 1H), 1.70 (s, 4H), 1.47 (d, J = 13.4 Hz, 4H), 1.31 (dd, J = 13.5, 6.9 Hz, 6H), 1.18 (m, 10H), 1.05 (s, 6H), 0.99 (m, 4H), 0.89 (d, J = 6.5 Hz, 3H), 0.82 (d, J = 12.6 Hz, 1H), 0.76 (s, 3H), 0.63 (s, 3H).

Embodiment 14

Compound 14 Preparation of 1,1,1-trifluoro-N-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]methanesulfonamide.

Step 1: In a 50 mL round-bottom flask, 3-amino-5α-cholest-25-ol 12 (20 mg, 0.050 mmol) was dissolved in dichloromethane (1 mL), triethylamine (15 mg, 0.15 mmol) was added at room temperature, trifluoromethanesulfonic anhydride (17 mg, 0.059 mmol) was added dropwise at 0°C, and the reaction solution was stirred at 0°C for 1 hour. The reaction was monitored by TLC (ethyl acetate/petroleum ether = 3:1). After the reaction, water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (5 mL×3). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated. The organic phase was purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1) to obtain a white solid 1,1,1-trifluoro-N-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]methanesulfonamide 14 (5 mg, purity: 91.44%, yield: 17.23%).

Compound 14: ¹ H NMR(400MHz,)δ4.53(d,J＝8.7Hz,1H),3.46(d,J＝8.1Hz,1H),1.94(dd,J＝17.4,13.9Hz,2H),1.77(dd,J ＝14.2,11.3Hz,2H),1.66( d,J＝10.3Hz,2H),1.45(d,J＝4.9Hz,3H),1.33(m,15H),1.21(s,6H),1.05(m,7H),0.91(d,J＝6.5 Hz,3H),0.79(s,3H),0.65(s,3H). ¹⁹ F NMR (377MHz, CDCl3) δ-77.92 (s, 3F).

Embodiment 18

Preparation of Compound 18 3β-[(3-hydroxypropyl)oxy]-5α-cholest-25-ol

Step 1: Dissolve 5α-cholestane-3β,25-diol 9-2 (200 mg, 0.494 mmol) in toluene (10 mL), add potassium tert-butoxide (277.28 mg, 2.471 mmol), and stir at room temperature for 1 hour. Add allyl bromide (298.95 mg, 2.471 mmol) and stir at room temperature for 2 hours. Monitor by TLC (petroleum ether: ethyl acetate = 3:1). The reaction solution is directly concentrated to obtain 40 mg of crude product, which is purified by column chromatography (ethyl acetate: petroleum ether = 30%) to obtain 3β-(prop-2-enyloxy)-5α-cholestane-25-ol 18-1 (70 mg, 0.150 mmol, 30.26%) as a white solid. ¹ H NMR(399MHz,Chloroform-d)δ5.91(ddt,J＝16.2,10.7,5.6Hz,1H),5.29–5.09(m,2H),3.99(dd,J＝5.6,1.7Hz,2H), 3.25(dt,J＝11.2,6.1Hz,1H),1.94(d,J＝12.4Hz,1H),1.87–1.74(m,2H),1.74–1.57(m,4H),1.50(d,J＝15.1Hz,4H) ,1.47–1.39(m,4H),1.36(d,J＝9.0Hz,4H),1.31(s,3H),1.19(s,6H),1.08(d,J＝8.3Hz,2H),1.03(d,J＝9.1Hz,3H), 0.95(d,J＝10.6Hz,2H),0.89(d,J＝6.4Hz,3H),0.87–0.79(m,2H),0.77(s,3H),0.62(s,3H),0.57(d,J＝13.1Hz,1H).

Step 2: Dissolve 3β-(prop-2-enyloxy)-5α-cholest-25-ol 18-1 (60 mg, 0.135 mmol) in tetrahydrofuran (3 mL), add borane tetrahydrofuran (1.349 mL, 1.349 mmol), and stir at room temperature for 1 hour. Add sodium hydroxide (0.899 mL, 2.698 mmol) and hydrogen peroxide (152.94 mg, 1.349 mmol), and stir at room temperature for 2 hours. Monitor by TLC (petroleum ether: ethyl acetate = 1:1). The reaction solution is directly concentrated to obtain 400 mg of crude product, which is purified by column chromatography (ethyl acetate: petroleum ether = 50%) to obtain 3β-[(3-hydroxypropyl)oxy]-5α-cholest-25-ol 18 (20 mg, 0.041 mmol, 30.43%) as a white solid.

Compound 18: ¹ H NMR (400 MHz, Chloroform-d) δ 3.76 (t, J = 5.4 Hz, 2H), 3.65 (tq, J = 5.5, 3.2 Hz, 2H), 3.21 (dt, J = 11.1, 6.0 Hz, 1H), 1.94 (dt, J = 12.8, 3.3 Hz, 1H), 1.86–1.82 (m, 1H), 1.81–1.78 (m, 2H), 1.77–1.66 (m, 3H), 1.62 (ddd, J = 12.6, 6.5, 3.1 Hz ,2H),1.57–1.46(m,2H),1.42(d,J＝11.5Hz,2H),1.38–1.30(m,5H),1.27–1.22(m,4H),1.19(s,6H),1.13–1.06(m, 2H),1.05–0.91(m,5H),0.89(d,J＝6.4Hz,3H),0.82(d,J＝8.4Hz,1H),0.77(s,3H),0.62(s,3H),0.60–0.54(m,1H). ^13C NMR (100MHz, cdcl3) δ79.053,77.192,71.095,67.771,62.813,56.447,56.162,54.332,44.758,44.385,42.577,39.988,36.886,36.389,35. 721,35.441,34.722,32.068,29.327,29.168,28.789,28.240,28.199,24.174,21.191,20.738,18.609,12.267,12.043. HPLC: 92.13% (CAD).

.

Embodiment 19

Preparation of compound 19 {[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetic acid.

Step 1: Dissolve {[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetate-2-methylpropan-2-yl ester (140 mg, 0.270 mmol) in dichloromethane (3 mL), add formic acid (6 mL, 0.010 mmol), and stir at room temperature for 1 hour. Monitor by TLC (petroleum ether:ethyl acetate=3:1). The reaction solution was directly concentrated to obtain crude {[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetic acid 19 (140 mg, 0.121 mmol, 44.85%) as a light yellow solid. Compound 19: ¹ H NMR (400MHz, Methanol-d4) δ4.08 (s, 2H), 3.33 (t, J = 5.0Hz, 1H), 2.01–1.96 (m, 1H), 1.84 (d, J＝13.6Hz,2H),1.76–1.61(m,3H),1.58–1.48(m,2H),1.42(d,J＝8.9Hz,3H),1.35(d,J＝20.6H z,4H),1.29(d,J＝9.8Hz,5H),1.21(s,1H),1.14(s,6H),1.13–1.06(m,4H),1.04–0.97(m,3H ),0.92(d,J＝6.6Hz,3H),0.87(s,1H),0.82(s,3H),0.67(s,3H),0.65–0.59(m,1H).LCMS:[M] ⁺ =462.37.

Embodiment 23

Preparation of Compound 23 3β,25-dihydroxycholest-4β-carboxylic acid

Step 1: In a 50 mL round-bottom flask at room temperature, acetic acid-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methyl ester 13 (220 mg, 0.46 mmol) was dissolved in dichloromethane (10 mL), and imidazole (157 mg, 2.31 mmol) and tert-butyldimethylsilyl chloride (348 mg, 2.31 mmol) were added at room temperature, followed by stirring at room temperature for 16 hours. The reaction was monitored by TLC (petroleum ether:ethyl acetate=5:1). After the reaction was completed, the reaction was quenched with water (50 mL), extracted with ethyl acetate (30 mL×3), the organic phase was washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain white solid acetic acid-[(1R, 3aS, 3bS, 5aS, 7S, 9aR, 9bS, 11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methyl ester 23-1 (180 mg, purity: 90%, yield: 59.40%). ¹ H NMR(400MHz, CDCl3)δ=4.24–4.05(m,2H),3.71–3.59(m,1H),1.95(s,3H),1.82–1.20(m,25H),1.17(s, 6H),1.10–0.93(m,5H),0.87(d,J＝6.5,3H),0.84(s,9H),0.69(s,3H),0.59(s,3H),0.00(d,J＝1.3,6H).

Step 2: In a 50 mL round-bottom flask, acetic acid-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methyl ester 23-1 (140 mg, 0.24 mmol) was dissolved in tetrahydrofuran (6 mL), and 1 mol/L lithium aluminum tetrahydride tetrahydrofuran solution (0.28 mL, 0.28 mmol) was added at room temperature, and stirred at room temperature for 0.5 hours. The reaction was monitored by TLC (ethyl acetate/petroleum ether = 3:1). Reaction results After completion, the reaction was quenched with water (50 mL), extracted with ethyl acetate (30 mL×3), the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give a white solid 3β-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-4-(hydroxymethyl)-5α-cholest-25-ol 23-2 (100 mg, purity: 90%, yield: 69.21%). ¹ H NMR(400MHz, CDCl3)δ=4.02(t,J=10.1,1H),3.95–3.87(m,1H),3.58(d,J=10.8,1H),2.15–1.63(m,16H),1.62–1.3 2(m,14H),1.25(s,9H),1.21(s,6H),1.13–0.94(m,7H),0.91(d,J＝6.5,3H),0.68(s,3H),0.63(s,3H),0.00(s,6H).

Step 3 In a 50 mL round-bottom flask, 3β-{[dimethyl(2-methylpropan-2-yl)silyl]oxy}-4-(hydroxymethyl)-5α-cholest-25-ol 23-2 (50 mg, 0.091 mmol) was dissolved in tetrahydrofuran (1.5 mL) and dichloromethane (1.5 mL), and Dess-Martin reagent (58 mg, 0.14 mmol) was added at room temperature, and stirred at room temperature for 2 hours. The reaction was monitored by TLC (ethyl acetate/petroleum ether = 4:1). After the reaction, the reaction was quenched with water (20 mL), extracted with ethyl acetate (10 mL×3), the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1) to give a white solid 3β-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-25-hydroxy-5α-cholestane-4-carboxaldehyde 23-3 (15 mg, purity: 90%, yield: 27.10%). ¹ H NMR(400MHz, CDCl3)δ=9.99(s,1H),3.60–3.53(m,1H),2.61–2.56(m,1H),2.03–1.93(m,2H),1.91–1.72(m,8H),1.69– 1.40(m,12H),1.26(s,9H),1.21(s,6H),1.13–0.99(m,8H),0.91(d,J＝6.3,3H),0.76(s,3H),0.64(s,3H),0.00(s,6H).

Step 4: In a 50 mL round-bottom flask at room temperature, 3β-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-25-hydroxycholest-4β-carboxaldehyde 23-3 (12 mg, 0.022 mmol) was dissolved in tert-butyl alcohol (1 mL) and tetrahydrofuran (0.5 mL), and 2-methylbut-2-ene (10 mg, 0.13 mmol), sodium dihydrogen phosphate (8 mg, 0.066 mmol), sodium chlorite (7 mg, 0.077 mmol) and water (0.25 mL) were added at 0°C, followed by stirring at room temperature for 16 hours. The reaction was monitored by TLC (dichloromethane: methanol = 10: 1). After the reaction, the reaction mixture was quenched with saturated aqueous sodium sulfite solution (20 mL), extracted with dichloromethane and methanol (dichloromethane: methanol = 10: 1, 10 mL × 3), and the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to give a white solid 3β,25-dihydroxycholest-4β-carboxylic acid 23 (3.65 mg, purity: 100%, yield: 37.09%).

Compound 23: ¹ H NMR (400 MHz, DMSO) δ＝11.86 (s, 1H), 4.49 (s, 1H), 4.03 (s, 1H), 3.50–3.43 (m, 1H), 2.55 (d, J＝5.2, 1H), 2.25–2.03 (m, 2H), 1.92 (d, J＝12.0, 1H), 1.87–1.69 (m, 2H), 1.65 (d, J＝13.0, 2H), 1.58–1.19 (m, 21H), 1.05 (s, 6H), 0.88 (d, J＝6.4, 3H), 0.83 (s, 3H), 0.61 (s, 1H). ¹³ C NMR (101MHz, DMSO) δ=71.37,69.23,56.19,55.54,54.29,52.79,50.66,47.76,44.60,42.52,38.01,36.64,36.13,35.69,35. 46,32.81,31.76,29.88,29.70,28.28,28.01,27.88,24.33,23.18,21.75,20.94,20.72,18.98,13.22,12.32.LC-MS:[M+H-2H ₂ O] ⁺ = 413.35

Embodiment 24

Compound 24 Preparation of 24,24-difluorocholest-6(5)-ene-3β,25-diol.

In the first step, zinc (0.61 g, 9.312 mmol) was dissolved in THF (15 mL), and then refluxed and stirred for 1 hour under nitrogen protection. Acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-formylpropan-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopentane A mixture of (1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-formylpropan-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2 _{-i]phenanthrene-7-yl acetate I-5 (1.2 g, 3.104 mmol) and BrCF2CO2Et (} 1.89 g, 9.312 mmol) was dissolved in THF (3 mL) and then added dropwise to the above zinc solution. The reaction was refluxed for 0.5 h. The reaction of the raw material was completed as monitored by TLC (petroleum ether:ethyl acetate = 5:1). The reaction solution was poured into water and extracted with ethyl acetate (20 mL x 2). The organic phases were combined and concentrated. The crude product was separated and purified by flash chromatography (petroleum ether: ethyl acetate = 95:5 to 80:20) to obtain a white solid (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluoro-3-hydroxyhexanoic acid ethyl ester 24-1 (700 ^mg , purity: 90%, yield: 39.74%). NMR(400MHz, CDCl3)δ5.37(d,J＝4.5Hz,1H),4.68–4.52(m,1H),4.47–4.28(m,2H),4 .12(d,J＝7.0Hz,1H),2.32(d,J＝7.1Hz,2H),2.08–1.94(m,5H),1.86(d,J＝9.6Hz,3H ),1.69(d,J＝13.0Hz,2H),1.60(td,J＝16.9,6.6Hz,4H),1.47(dd,J＝15.7,11.6Hz,3 H),1.37(t,J＝7.1Hz,3H),1.29–1.09(m,6H),1.07–0.94(m,8H),0.75–0.67(m,3H).

In the second step, (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluoro-3-hydroxyhexanoic acid ethyl ester 24-1 (300 mg, 0.587 mmol) was dissolved in dichloromethane (10 mL), and then di(imidazole-1-yl)methanone (314.08 mg, 1.762 mmol) and DMAP (14.35 mg, 0.117 mmol) were added in sequence. The reaction was refluxed for 12 hours and monitored by TLC (petroleum ether: ethyl acetate = 3:1). The reaction solution was poured into water and extracted with ethyl acetate (20 mL x 2). The organic phases were combined and concentrated. The crude product was separated and purified by flash chromatography (petroleum ether: ethyl acetate = 95:5 to 60:40) to obtain a colorless oily liquid (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluoro-3-{[(imidazol-1- ^yl )sulfanylidenemethyl]oxy}hexanoic acid ethyl ester 24-2 (250 mg, purity: 90%, 61.69%). NMR (400MHz, CDCl3) δ8.47(d,J＝9.6Hz,1H),7.67(d,J＝11.0Hz,1H),7.14(d,J＝6.8Hz,1 H),6.32–6.12(m,1H),5.37(d,J＝4.8Hz,1H),5.30(s,2H),4.68–4.54(m,1H),4.32(q,J ＝7.2Hz,2H),2.32(d,J＝6.8Hz,2H),2.11(s,1H),2.06–1.93(m,5H),1.85(d,J＝10.4Hz, 3H),1.73–1.39(m,9H),1.35–1.24(m,5H),1.22–0.94(m,13H),0.68(t,J=18.8Hz,3H).

Step 3: (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluoro-3-{[(imidazol-1-yl)sulfanylidenemethyl]oxy}hexanoic acid ethyl ester 24-2 (200 mg, 0.322 mmol) was dissolved in toluene (5 mL), and benzoic acid peroxyanhydride (15.61 mg, 0.064 mmol) and triethylsilyl hydrochloride (299.69 mg, 2.577 mmol) were added in sequence, and the reaction was reacted under reflux for 18 hours. TLC (petroleum ether:ethyl acetate = 8:1) monitored the reaction to be complete. The reaction solution was poured into water and extracted with ethyl acetate (20 mL x 2). The organic phases were combined and concentrated. The crude product was separated and purified by flash chromatography (petroleum ether: ethyl acetate = 95:5) to obtain (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluorohexanoic acid ethyl ester 24-3 (90 mg, purity: 90%, 50.83%), which was directly used for the next step.

Step 4: Dissolve (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluorohexanoic acid ethyl ester 24-3 (70 mg, 0.142 mmol) in THF (4 mL), cool to 0 ^° C, slowly add 3.0 M MeMgBr (0.28 mL) dropwise, and stir at 0°C for 0.5 h after the addition is complete. After the reaction was completed as monitored by TLC (petroleum ether: ethyl acetate = 5:1), the mixture was quenched with water (10 mL), extracted with ethyl acetate (10 mL x 2), and concentrated. The crude product was separated and purified by flash chromatography (petroleum ether: ethyl acetate = 95:5 to 90:10) to give white solid acetate-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 24-4 (50 mg, 90%, 69.83%). ¹ H NMR (400MHz, CDCl3) δ5.46–5.27(m,1H),4.76–4.47(m,1H),2.33(s,2H),2.08–1.95(m,5H),1.86(d,J＝12.4Hz,3H),1.51(d t,J＝12.4,7.9Hz,13H),1.31(s,8H),1.14(s,3H),1.02(s,4H),0.94(d,J＝6.6Hz,3H),0.87(d,J＝11.6Hz,1H),0.69(s,3H).

Step 5: Acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 24-4 (60 mg, 0.125 mmol) was dissolved in MeOH (5 mL) and water (1 mL), and then K ₂ CO ₃ (34.5 mg, 0.25 mmol) was added and the reaction was allowed to react at room temperature for 1 hour. TLC ( The reaction solution was poured into water and extracted with ethyl acetate (20 mL x 2). The organic phases were combined and concentrated. The crude product was separated and purified by flash chromatography (petroleum ether: ethyl acetate = 95:5-60:40) to obtain a white solid 24,24-difluorocholest-6(5)-ene-3β,25-diol 24 (30 mg, 0.065 mmol, 52.05%).

Compound 24: ^1H NMR (400MHz, CDCl3) δ5.35(d,J＝5.2Hz,1H),3.52(dd,J＝10.3,5.6Hz,1H),2.34–2.18(m,2H),2.07–1.92(m,3H ),1.86(dt,J＝13.9,4.8Hz,3H),1.74–1 .62(m,2H),1.49(ddt,J＝18.0,14.7,8.8Hz,7H),1.37–1.24(m,9H),1.18– 1.04(m,4H),1.05–0.98(m,4H),0.97–0.89(m,4H),0.67(d,J=12.8Hz,3H). ^13C NMR (101MHz, CDCl3) δ140.76,121.70,71.81,56.72,55.71,50.01,42.36,42.30,39.75,37.21,36.46,35. 37,31.90,31.89,31.66,28.03,27.32,26.87,24.12,23.60,21.08,19.40,18.46,11.79.LC-MS: [M+H-H2O] ⁺ =421.55

Embodiment 25

Compound 25 Preparation of 24,24-difluoro-5α-cholest-3β,25-diol.

In the first step, the obtained white solid acetate-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 24-4 (40 mg, 0.083 mmol) was dissolved in methanol (4 mL), and Pd/C (40 mg) was added. After the addition was completed, the mixture was stirred at room temperature for 3 hours. After the reaction was complete as monitored by TLC (petroleum ether: ethyl acetate = 2:1), the mixture was filtered and the crude product was directly used for the next step.

In the second step, (1R, 3aS, 3bR, 5aS, 7S, 9aS, 9bS, 11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl acetate 25-1 (40 mg, 0.083 mmol) was dissolved in methanol (5 ml) and water (0.5 ml), and then potassium carbonate (34.36 mg, 0.249 mmol) was added. The reaction was allowed to react at room temperature for 1 hour, and new spots were generated by monitoring TLC (petroleum ether: ethyl acetate = 3:1). The reaction solution was poured into water and extracted with ethyl acetate (20 ml x 2). The organic phases were combined and concentrated. The crude product was separated and purified by flash chromatography (petroleum ether: ethyl acetate = 95:5-60:40) to obtain a white solid 24,24-difluoro-5α-cholest-3β,25-diol 25 (10 mg, 0.020 mmol, 24.65%). Compound 25: ¹ H NMR(400MHz, CDCl3) δ3.59(s,1H),1.95(d,J＝12.6Hz,2H),1.71(ddd,J＝27.4,24.6,6.5Hz,6H),1.53–1.42(m, 4H),1.41–1.22(m,14H),1.17–0.95(m,6H),0.91(t,J＝8.9Hz,4H),0.80(s,3H),0.66(s,4H).LC-MS: [M+H-H2O] ⁺ =423.55

Embodiment 26

Compound 26 Preparation of 4β-hydroxy-3β-hydroxy-24-(hydroxycyclopropyl)-5α-cholan-7-one.

Step 1: Dissolve (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-8 (300 mg, 0.65 mmol) in dichloromethane (20 mL), add 4-dimethylaminopyridine (39.81 mg, 0.03 mmol), triethylamine (197.70 mg, 1.95 mmol) and acetic anhydride (79.78 mg, 0.78 mmol) and react at room temperature for 2 h. TLC (petroleum ether: ethyl acetate = 5:1) monitors the reaction to be complete. The residue was washed with dichloromethane and water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate ~ 20:1) to obtain (5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-diacetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro- 1H-Cyclopenta[1,2-a]phenanthren-1-yl]hexanoic acid methyl ester 26-1 (230 mg, 0.441 mmol, 62.2%) white solid. ¹ H NMR(399MHz,Chloroform-d)δ5.81–5.76(m,1H),5.48(d,J＝3.4Hz,1H),3.64(s,3H),2.35–2.18(m,2H),2.04(s,3H),1.99(s,3H),1.87(dt,J＝13.8 ,3.7Hz,2H),1.56(dd,J＝6.7,4.2Hz,2H),1.50–1.42(m,4H),1.39–1.33(m ,2H),1.24–1.17(m,2H),1.11(s,3H),0.91(d,J＝6.4Hz,3H),0.65(s,3H).

Step 2: Dissolve (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 26-1 (100.00 mg, 0.23 mmol) in acetone (3.0 mL), nitrogen protection, N-hydroxyphthalimide (3.25 mg, 0.02 mmol), cobalt acetate (0.35 mg, 0.01 mmol) and then add tert-butyl peroxide (85 mg, 0.90 mmol), and react at room temperature for 16 h. TLC (petroleum ether: ethyl acetate = 10:1) monitored the reaction to be complete. Sodium thiosulfate was added to quench, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and column chromatographed with petroleum ether:ethyl acetate (100% to 20%) to obtain (5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-diacetoxy-9a,11a-dimethyl-4-oxyylidene-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl] ^hexanoic acid methyl ester 26-2 (40 mg, 0.07 mmol, 35.0%). NMR(399MHz,Chloroform-d)δ5.90(s,1H),5.60–5.56(m,1H),4.80–4.74(m,1H),3. 64(d,J＝0.8Hz,3H),2.38–2.22(m,4H),2.07(d,J＝0.8Hz,3H),2.06–2.02(m,1H),2.0 0(s,3H),1.98–1.85(m,2H),1.77(dd,J＝13.0,4.0Hz,1H),1.47–1.32(m,3H),1.29( s,3H),1.23(d,J＝8.5Hz,2H),1.14–1.04(m,3H),0.92(d,J＝6.5Hz,3H),0.65(s,3H).

Step 3: Dissolve (5R)-5-[(1R, 3aS, 3bS, 6R, 7S, 9aR, 9bS, 11aR)-6,7-diacetoxy-9a, 11a-dimethyl-4-oxyylidene-2,3,3a, 3b, 4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 26-2 (300 mg, 0.65 mmol) in methanol (10.0 mL), Pd/C 10% (20 mg, 0.19 mmol), purify with hydrogen three times, react at room temperature for 1 h, and monitor the completion of the reaction by TLC (petroleum ether: ethyl acetate = 5:1). Filtered with celite, rinsed with methanol, concentrated, and column chromatographed with petroleum ether:ethyl acetate (100% to 20%), to obtain (5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-diacetoxy-9a,11a-dimethyl-4-oxyylidenehexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 26-3 (50 mg) as a white solid, which was directly used in the ^next step. NMR (399MHz, Chloroform-d) δ5.15 (s, 1H), 4.76 (dt, J = 12.3, 4.2Hz, 1H), 3.65 (s, 3H), 2.49 (t, J = 13. 5Hz,1H),2.33–2.16(m,4H),2.09(s,3H),2.06(dd,J＝13.0,3.1Hz,1H),1.96(s,3H),1.92(d,J＝3.7H z,1H),1.90–1.80(m,3H),1.74–1.63(m,3H),1.49(dd,J＝9.8,6.4Hz,3H),1.37(td,J＝11.7,11.3,7. 0Hz,3H),1.26(s,3H),1.17(d,J＝12.5Hz,2H),1.12–1.03(m,5H),0.91(d,J＝6.4Hz,3H),0.62(s,3H).

Step 4: Dissolve (5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-diacetoxy-9a,11a-dimethyl-4-oxyylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 26-3 (50 mg, 0.10 mmol) in toluene (5 mL), add ethylene glycol (0.05 mL, 0.96 mmol), anhydrous p-toluic acid (7.3 mg, 0.05 mmol), heat under reflux and stir for 1 h. Monitor the reaction by TLC (petroleum ether: ethyl acetate = 2:1). Water was added, extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, concentrated and column chromatographed with petroleum ether:ethyl acetate (100% to 20%) to obtain (5R)-5-[(1'R,3a'S,3b'S,6'R,7'S,9a'R,9b'S,11a'R)-6',7'-diacetoxy-9a',11a'-dimethyl-1',2',3',3a',3b',5',5a',6',7',8',9',9a',9b',10',11',11a'-hexahydrospiro[1,3-dioxolane-2,4'-cyclopenta[1,2-a] [phenanthrenyl]-1'-yl] hexanoic acid methyl ester 26-4 (8 mg, 0.01 mmol, 17.2%) white solid. ¹ H NMR (399 MHz, Chloroform-d) δ 5.14 (s, 1H), 4.76 (dt, J = 12.0, 4.3 Hz, 1H), 3.97–3.84 (m, 5H), 3.63 (s, 3H), 2.05 (s, 3H), 1.94 (s, 2H), 1.02 (s, 3H), 0.90 (d, J = 6.5 Hz, 3H), 0.62 (s, 3H)

Step 5: Dissolve compound (5R)-5-[(1'R,3a'S,3b'S,6'R,7'S,9a'R,9b'S,11a'R)-6',7'-diacetoxy-9a',11a'-dimethyl-1',2',3',3a',3b',5',5a',6',7',8',9',9a',9b',10',11',11a'-hexahydrospiro[1,3-dioxolane-2,4'-cyclopenta[1,2-a]phenanthrene]-1'-yl]hexanoic acid methyl ester 26-4 (20 mg, 0.04 mmol) in tetrahydrofuran (5 mL), add tetraisopropyl titanate (95.6 mg, 0.67 mmol) at room temperature, purify with nitrogen three times, cool to 0°C and add ethyl magnesium chloride (0.62 mL) dropwise. The mixture was stirred at room temperature for 1 h and monitored by TLC (petroleum ether:ethyl acetate=3:1). The mixture was quenched by adding brine at low temperature, diluted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography with petroleum ether:ethyl acetate (100%-50%) to give (1'R,3a'S,3b'S,5a'R,6'R,7'S,9a'R,9b'S,11a'R)-1'-[(2R)-5-(hydroxycyclopropyl)pentan-2-yl]-9a',11a'-dimethyl-1',2',3',3a',3b',5',5a',6',7',8',9',9a',9b',10',11',11a'-hexahydrospiro[1,3-dioxolane-2,4'-cyclopenta[1,2-a]phenanthrene]-6',7'-diol 26-5 (15.0 mg, 0.03 mmol, 33.6%) as a white solid. ¹ H NMR(399MHz,Chloroform-d)δ3.96(d,J＝5.9Hz,4H),3.67(s,1H),1.94(d,J＝12.9Hz,2H),1.80(d,J＝13.0Hz,5H),1.73(d,J＝10.9Hz,5H),1.4 4(d,J＝7.8Hz,6H),1.35(s,8H),1.24(s,3H),1.04(s,3H),0.92(d,J＝6 .2Hz,3H),0.84(t,J＝7.8Hz,4H),0.71(s,2H),0.64(s,3H),0.42(s,2H)

Step 5: Dissolve (1'R,3a'S,3b'S,5a'R,6'R,7'S,9a'R,9b'S,11a'R)-1'-[(2R)-5-(hydroxycyclopropyl)pentan-2-yl]-9a',11a'-dimethyl-1',2',3',3a',3b',5',5a',6',7',8',9',9a',9b',10',11',11a'-hexahydrospiro[1,3-dioxolane-2,4'-cyclopenta[1,2-a]phenanthrene]-6',7'-diol 26-5 (10 mg, 0.02 mmol) in methanol (3.0 mL), add acetic acid (2 mL, 34.94 mmol) at room temperature, stir at room temperature for 1 h, and monitor the reaction completion by TLC (petroleum ether:ethyl acetate = 1:1). The mixture was quenched by adding brine at low temperature, diluted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography with petroleum ether:ethyl acetate (100%-50%) to give 4β-hydroxy-3β-hydroxy-24-(hydroxycyclopropyl)-5α-cholan-7-one 26 (3 mg, 0.01 mmol, 19.3%).

Compound 26: ¹ H NMR (399 MHz, Chloroform-d) δ 3.70 (s, 1H), 3.57 (d, J = 10.6 Hz, 1H), 2.86 (t, J = 13.5 Hz, 1H), 2.37 (t, J = 11.3 Hz, 1H), 2.21 (d, J = 12.1 Hz, 1H), 2.14–2.02 (m, 2H), 1.97 (d, J = 13.1 Hz, 1H), 1.79 ( ddt,J＝45.8,32.3,14.7Hz,4H),1.58–1.38(m,9H),1.27(s,6H),1.11–0.96(m,6iH),0.91(t,J =6.0Hz,3H),0.84(t,J＝7.4Hz,2H),0.71(d,J＝5.4Hz,2H),0.63(s,3H),0.43(t,J＝3.3Hz,2H). ^13C NMR(101MHz,cdcl3)δ212.83,77.31,77.20,76.99,76.67,74.71,73.76,7 1.92,55.93,55.76,54.82,50.03,48.95,43.78,42.51,38.80,38.60,36.5 2,36.06,35.89,35.88,35.62,35.55,31.03,29.68,28.40,25.58,24.99, 22.29,21.09,18.74,14.16,13.57,13.48,12.04,7.80,1.00.LC-MS:[M+H] ⁺ =433.3.

Embodiment 27

Compound 27 Preparation of 3-{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}propanoic acid.

Step 1: Dissolve 3β-[(3-hydroxypropyl)oxy]-5α-cholest-25-ol 18 (70 mg, 0.151 mmol) in dichloromethane (5 mL), add Dess-Martin periodinane (64.16 mg, 0.151 mmol), and stir at room temperature for 1 hour. Monitor by TLC (petroleum ether: ethyl acetate = 1:1). The reaction solution is directly concentrated to obtain 100 mg of crude product, which is purified by column chromatography (ethyl acetate: petroleum ether = 30%) to obtain 3-{[(1R, 3aS, 3bR, 5aS, 7S, 9aS, 9bS, 11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}propanal 27-1 (50 mg, 0.103 mmol, 68.15%) as a white solid. ¹ H NMR(399MHz,Chloroform-d)δ9.77(t,J＝2.0Hz,1H),3.78(td,J＝6.3,3.1Hz,2H),3.22(dt,J＝11.5,6.2H z,1H),2.63(td,J＝6.2,2.0Hz,2H),1.94(d,J＝12.5Hz,1H),1.81(d,J＝13.9Hz,2H),1.73–1.60(m,3H),1. 58–1.44(m,3H),1.36(q,J＝13.9,12.9Hz,5H),1.12(d,J＝26.3Hz,2H),1.04(t,J＝9.5Hz,3H),0.96(d,J＝ 20.8Hz,2H),0.89(d,J＝6.3Hz,3H),0.82(dd,J＝16.9,8.3Hz,2H),0.76(s,3H),0.62(s,3H),0.59(s,1H).

Step 2: 3-{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}propanal 27-1 (45 mg, 0.098 mmol) was dissolved in tert-butyl alcohol (4 mL) and water (0.5 mL), sodium dihydrogen phosphate (43.19 mg, 0.360 mmol), 2-methyl-2-butene (137.01 mg, 1.953 mmol), sodium chlorite (26.50 mg, 0.293 mmol) were added, and the mixture was stirred at room temperature for 1 hour. TLC (dichloromethane: methanol = 20:1) was used for monitoring. The mixture was diluted with water and extracted three times with ethyl acetate. The organic phase was washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain 20 mg of solid. The solid was purified by column chromatography (methanol/dichloromethane = 10%) to obtain 3-{[(1R, 3aS, 3bR, 5aS, 7S, 9aS, 9bS, 11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}propanoic acid 27 (24 mg, 0.048 mmol, 48.97%) as a white solid. Compound 27: ¹ H NMR (400 MHz, Methanol-d ₄ )δ3.71(t,J＝6.3Hz,2H),3.25(d,J＝10.9Hz,1H),2.48(t,J＝6.3Hz,2H),2.02–1.95(m,1H),1.88–1.77(m,2H ),1.76–1.64(m,2H),1.64–1.55(m,2H),1.50(dd,J＝13.2,4.1Hz,1H),1.42(d,J＝8.9Hz,2H),1.38(t,J＝4.0H z,2H),1.35–1.32(m,2H),1.27(t,J＝12.4Hz,5H),1.19(d,J＝10.9Hz,2H),1.14(s,6H),1.13–1.07(m,3H),1. 07–0.94(m,4H),0.92(d,J＝6.6Hz,3H),0.90–0.83(m,1H),0.80(s,3H),0.67(s,3H),0.62(d,J＝16.2Hz,1H). ^13C NMR(101MHz,cd3od) δ174.242,78.790,70.026,63.304,56.451,56.228,54.382,44.646,43.843,42.334,39.970,36.684,36.350,35.704,35.429,35.3 78,34.873,34.432,31.875,28.548,27.897,27.806,27.780,27.664,23.804,20.894,20.416,17.736,11.265,11.060.LCMS:[Ms-H] ^- =475.45.

Embodiment 29

Preparation of Compound 29 3β-[(2-hydroxyethyl)oxy]-5α-cholest-4β,25-diol

Step 1: Weigh acetic acid-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-methoxy-6-oxydehex-2-yl]-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester I-7 (77 mg, 0.17 mol, 1.0 eq.) and dissolve it in tetrahydrofuran (2 mL). After nitrogen replacement, dry ice -Acetone was cooled to -78°C, methyl lithium (0.54 mL, 0.87 mmol) was added, and then the temperature was raised to room temperature and stirred. TLC (petroleum ether: ethyl acetate = 10:1) was used for monitoring. After the reaction was completed, 6 mL of water was added to quench the reaction, and 5 mL of ethyl acetate was used for extraction. The organic phase was dried and concentrated. The crude product was purified by column chromatography (dichloromethane: methanol = 300: ¹ to 100:1) to obtain cholest-5(6)-ene-3β,25-diol 29-1 (35 mg, 0.083 mmol, 47.66%) as a white solid. HNMR(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). ^13C 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.

Step 2: Cholest-5(6)-ene-3β,25-diol 29-1 (1280 mg, 3.179 mmol, 1.0 eq.) was dissolved in toluene (50 mL), potassium tert-butoxide (3567.00 mg, 31.789 mmol, 10.0 eq.) was added, and stirred at room temperature for 1 hour. Then tert-butyl bromoacetate (6200.69 mg, 31.789 mmol, 10.0 eq.) was added, and stirred at room temperature for 2 hours. TLC (petroleum ether: ethyl acetate = 3:1) was monitored. The reaction solution was directly concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate: petroleum ether = 30%) to obtain {[(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-tetradecahydro ^- 1H-cyclopenta[1, 2-a] phenanthren-7-yl] oxy} acetate-2-methylpropan-2-yl ester 29-2 (950 mg, 1.746 mmol, 54.93%) as a white solid. NMR(399MHz,Chloroform-d)δ5.33(d,J＝5.0Hz,1H),3.98(s,2H),3.27–3.15(m,1H),2.4 2–2.33(m,1H),2.24(t,J＝12.2Hz,1H),1.95(q,J＝12.5Hz,3H),1.86–1.73(m,2H),1.62– 1.50(m,3H),1.48(s,2H),1.45(s,9H),1.43–1.41(m,3H),1.39–1.33(m,5H),1.19(s,7H ),1.15–1.08(m,2H),1.07–1.00(m,3H),0.98(s,3H),0.91(d,J＝6.5Hz,3H),0.65(s,3H).

Step 3: 2-methylpropan-2-yl {[(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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetate 29-2 (950 mg, 1.746 mmol, 54.93%) (950 mg, 1.838 mmol, 1.0 eq.) was dissolved in dichloromethane (15 mL), and pyridine (0.740 mL, 9.191 mmol, 5.0 eq.) and acetyl chloride (0.392 mL, 5.515 mmol, 3.0 eq.) were added. The mixture was stirred at 40°C for 5 hours. TLC (petroleum ether: ethyl acetate = 5:1) was used for monitoring. The reaction solution was directly concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate: petroleum ether = 20%) to obtain {[(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 -tetradecahydro-1H-cyclopenta[1, 2-a] phenanthren-7-yl] oxy}acetate-2-methylpropan-2-yl ester 29-3 (760 mg, 1.292 mmol, 70.28%) as a white solid. NMR(399MHz,Chloroform-d)δ5.33(s,1H),3.98(s,2H),3.26–3.16(m,1H),2.41–2.32(m,1H),2.24(t ,J＝12.4Hz,1H),2.00(d,J＝3.9Hz,1H),1.95(s,3H),1.88(d,J＝15.1Hz,1H),1.83–1.70(m,2H),1.68–1 .61(m,1H),1.59–1.49(m,4H),1.45(d,J＝1.8Hz,9H),1.40(s,6H),1.37–1.30(m,3H),1.29–1.17(m,2 H),1.17–1.09(m,2H),1.04(dd,J＝12.3,10.5Hz,3H),0.98(s,3H),0.89(d,J＝6.3Hz,3H),0.65(s,3H).

Step 4: 2-methylpropan-2-yl {[(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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetate 29-3 (760 mg, 1.360 mmol, 1.0 eq.) was dissolved in chloroform (5 mL), and N-methylmorpholine (825.36 mg, 8.160 mmol, 6.0 eq.) and selenium dioxide (198.55 mg, 1.789 mmol, 5.0 eq.) were added. The mixture was stirred at 75°C for 18 hours. The reaction solution was directly concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate: petroleum ether = 40%) to obtain {[(1R, 3aS, 3bS, 6R, 7S, 9aR, 9bS, 11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-6-hydroxy-9a, 11a-dimethyl-2, 3, 3a, 3b, 4, 6, 7, 8, 9, 9a, 9b, 10, 11, 11a-tetradecahydro-1H-cyclopenta[1, 2-a]phenanthrene-7-yl]oxy}acetate-2-methylpropan-2-yl ester 29-4 (320 mg, 0.529 mmol, 38.89%) as a white solid. ¹ H NMR(399MHz,Chloroform-d)δ5.68(d,J＝4.5Hz,1H),4.19–4.11(m,2H),3.93(d,J＝16.8 Hz,1H),3.26(d,J＝11.6Hz,1H),2.10–2.03(m,1H),1.99(d,J＝12.5Hz,2H),1.95(s,3H), 1.83(d,J＝14.2Hz,2H),1.66(d,J＝14.3Hz,3H),1.52(d,J＝11.1Hz,3H),1.46(s,9H),1.40(s,6H),1.34(d,J＝6.7Hz,2H),1.24(d,J＝8.7H z,1H),1.18(s,3H),1.12(t,J＝14.3Hz,2H),1.06–1.01(m,2H),1.00–0.93(m,2H),0.89(d,J＝6.4Hz,3H),0.88–0.79(m,1H),0.65(s,3H).

Step 5: Dissolve {[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-6-hydroxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetate-2-methylpropan-2-yl ester 29-4 (320 mg, 0.557 mmol, 1.0 eq.) in ethyl acetate (10 mL) and acetic acid (2 mL), and add platinum dioxide (160 mg, 0.705 mmol, 1.3 eq.). Stir at 40°C for 15 min under hydrogen atmosphere. Monitor by TLC (petroleum ether:ethyl acetate = 5:1). The reaction solution was filtered, and the filtrate was concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate: petroleum ether = 30%) to obtain {[(1R, 3aS, 3bS, 6R, 7S, 9aR, 9bS, 11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-6-hydroxy- ^9a , 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetate-2-methylpropan-2-yl ester 29-5 (200 mg, 0.329 mmol, 59.17%) as a white solid. NMR(400MHz,Chloroform-d)δ4.15(d,J＝16.6Hz,1H),3.90(d,J＝16.8Hz,1H),3.74(s,1H),3.22(d,J＝11.4Hz,1H),1.95 (d,J＝1.1Hz,3H),1.90(d,J＝12.8Hz,1H),1.80(d,J＝13.4Hz,2H),1.73(dd,J＝13.3,8.3Hz,4H),1.63(dd,J＝11.8,7.8Hz, 3H),1.59–1.48(m,2H),1.45(d,J＝1.2Hz,9H),1.39(d,J＝1.2Hz,6H),1.34(d,J＝3.7Hz,5H),1.27–1.21(m,3H),1.16–1.0 6(m,2H),1.02(s,3H),0.99–0.91(m,3H),0.88(d,J＝6.4Hz,3H),0.80(d,J＝14.0Hz,1H),0.63(s,3H),0.57–0.49(m,1H).

Step 6: Dissolve {[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-1-[(2R)-6-acetoxy-6-methylhept-2-yl]-6-hydroxy-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}acetate-2-methylpropan-2-yl ester 29-5 (40 mg, 0.069 mmol, 1.0 eq.) in tetrahydrofuran (5 mL), add lithium aluminum tetrahydride (13.16 mg, 0.347 mmol, 5.0 eq.). Stir at room temperature for 1 h. Monitor by TLC (petroleum ether: ethyl acetate = 5:1). The reaction solution was quenched with water, filtered, and the filtrate was concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate: petroleum ether = 50%) to obtain 3β-[(2-hydroxyethyl)oxy]-5α-cholest-4β,25-diol 29 (20 mg, 0.041 mmol, 58.96%) as a white solid. Compound 29: ¹ H NMR (399 MHz, Chloroform-d) δ 3.88 (s, 1H), 3.73 (q, J = 3.8, 3.3 Hz, 2H), 3.62 (t, J = 4.6 Hz, 2H), 3.25 (dt, J = 11.9, 4.3 Hz, 1H), 1.94 (d, J = 12.3 Hz, 1H), 1.83–1.73 (m, 5H), 1.70 (s, 6H), 1.54 (d, J = 9.1 Hz, 1H), 1.41 (s ,2H),1.36(d,J＝13.0Hz,5H),1.30–1.22(m,2H),1.19(s,6H),1.07(d,J＝5.7Hz,2H),1.02(s,3H),0.95(d ,J＝17.5Hz,2H),0.90(d,J＝6.5Hz,3H),0.88–0.80(m,1H),0.63(s,3H),0.60–0.51(m,1H).LCMS:[M-OH-H ₂ O] ⁺ = 429.40; CAD: 92.03%.

Embodiment 31

Compound 31 Preparation of 25-hydroxy-3β-[(2-hydroxyethyl)oxy]-5α-cholest-4-one.

Step 1: Dissolve 3β-[(2-hydroxyethyl)oxy]-5α-cholestane-4β,25-diol 29 (86 mg, 0.185 mmol, 1.0 eq.) in dichloromethane (5 mL), add imidazole (62.99 mg, 0.925 mmol, 5.0 eq.) and tert-butyldimethylsilyl chloride (83.67 mg, 0.555 mmol, 3.0 eq.). Stir at room temperature for 2 h. Monitor by TLC (petroleum ether: ethyl acetate = 5:1). The reaction solution was concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate: petroleum ether = 20%) to obtain 3β-[(4,4,5,5-tetramethyl-3-oxa-4-silanhexyl)oxy]-5α-cholestane-4β,25-diol 31-1 (65 mg, 0.107 mmol, 57.63%) as a white solid. ¹ H NMR(400MHz,Chloroform-d)δ3.84(s,1H),3.72(dq,J＝11.5,6.8,6.4Hz,2H),3.55(d ,J＝10.4Hz,2H),3.22(d,J＝11.1Hz,1H),1.93(d,J＝12.3Hz,1H),1.74(dt,J＝24.6,12 .8Hz,5H),1.36(s,5H),1.23(s,2H),1.19(s,6H),1.06(s,3H),1.01(s,3H),0.95(d, J＝14.0Hz,2H),0.91–0.87(m,12H),0.63(s,3H),0.54(d,J＝3.1Hz,1H),0.05(s,6H).

Step 8: Dissolve 3β-[(4,4,5,5-tetramethyl-3-oxa-4-silanhex-1-yl)oxy]-5α-cholest-4β,25-diol 31-1 (60 mg, 0.104 mmol, 1.0 eq.) in dichloromethane (1 mL), add Dess-Martin periodinane (43.95 mg, 0.104 mmol, 1.0 eq.). Stir at room temperature for 1 h. Monitor by TLC (petroleum ether: ethyl acetate = 5:1). The reaction solution is concentrated to obtain a crude product, which is purified by column chromatography (ethyl acetate: petroleum ether: ethyl acetate = 5:1). Oil ether = 20%) to give 25-hydroxy-3β-[(4,4,5,5-tetramethyl-3-oxa-4-silanahex-1-yl)oxy]-5α-cholest-4-one 31-2 (50 mg, 0.082 mmol, 79.44%) as a white solid. ¹ H NMR (399 MHz, Chloroform-d) δ 4.01 (dd, J = 12.1, 7.1 Hz, 1H), 3.83–3.73 (m, 3H), 3.44 (dt, J = 11.3, 5.2 Hz, 1H), 2.25 (s, 1H), 2.02 (dd, J = 38.0, 12.2 Hz, 2H), 1.83 (dd, J = 37.0, 11.2 Hz, 3H), 1.72 (d, J = 13.6 Hz, 3H), 1.50 (d, J = 2 0.7Hz,4H),1.38(dd,J＝18.2,9.9Hz,4H),1.25(d,J＝10.6Hz,3H),1.20(s,6H),1.15–1.06(m,2H),1.06–0.95( m,3H),0.91–0.88(m,3H),0.87(s,9H),0.78(d,J＝3.9Hz,1H),0.70(s,3H),0.63(s,3H),0.04(d,J＝2.8Hz,6H).

Step 9: Dissolve 25-hydroxy-3β-[(4,4,5,5-tetramethyl-3-oxa-4-silanhex-1-yl)oxy]-5α-cholest-4-one 31-2 (40 mg, 0.069 mmol, 1.0 eq.) in tetrahydrofuran (2 mL), add tetrabutylammonium fluoride (0.5 mL, 0.500 mmol, 7.2 eq.). Stir at room temperature for 1 h. Monitor by TLC (petroleum ether: ethyl acetate = 3:1). Add water to the reaction solution, extract with ethyl acetate three times, dry the organic phase and concentrate to obtain a crude product, which is purified by column chromatography (methanol/ethyl acetate = 5%) to obtain 25-hydroxy-3β-[(2-hydroxyethyl)oxy]-5α-cholest-4-one 31 (12 mg, 0.025 mmol, 35.54%) as a white solid.

Compound 31: ¹ H NMR (400MHz, Methanol-d4) δ4.08 (dd, J = 11.8, 7.2 Hz, 1H), 3.70–3.59 (m, 3H), 3.52 (dd, J = 9.3, 3.4 Hz, 1H), 2.35–2.22 (m, 2H), 2.20–1.96 (m, 2H), 1.95–1.77 (m, 2H), 1.76–1.61 (m, 2H), 1.55 (dq, J = 19.4, 6.4 Hz, 4H), 1.40 (d,J＝9.3Hz,3H),1.34–1.30(m,3H),1.27(d,J＝4.0Hz,3H),1.19(d,J＝12.7Hz,1H),1.15(s,6H),1.08(d,J＝1 3.2Hz,2H),1.03–0.96(m,2H),0.93(d,J＝6.4Hz,3H),0.90–0.80(m,2H),0.69(d,J＝11.8Hz,6H).LCMS:[M+H] ⁺ = 463.40; CAD: 80.15%.

Embodiment 33

Preparation of Compound 33 [(1R, 3aS, 3bS, 5aS, 7S, 9aR, 9bS, 11aR)-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]acetonitrile

Step 1: In a 50 mL round-bottom flask at room temperature, 4-(hydroxymethyl)-5α-cholest-3β,25-diol 13-1 (120 mg, 0.28 mmol) was dissolved in 1,2-dichloroethane (5 mL), and triethylamine (170 mg, 1.68 mmol) and 4-dimethylaminopyridine (34 mg, 0.28 mmol) and p-toluenesulfonyl chloride (107 mg, 0.56 mmol), and then stirred at 50°C for 8 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 2: 1). After the reaction was completed, it was quenched with water (30 mL), extracted with ethyl acetate (20 mL × 3), the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain a white solid 4-methylbenzenesulfonic acid-[(1R, 3aS, 3bS, 5aS, 7S, 9aR, 9bS, 11aR)-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methyl ester 33-1 (60 mg, purity: 90%, yield: 36.36%). ¹ HNMR (400MHz, CDCl3) δ = 7.80 (d, J = 8.2, 2H), 7.34 (d, J = 8.1, 2H), 4.33–4.22 (m, 1H), 4.19–4.11 (m, 1H), 3.79–3.71 (m, 1H), 2.45 (s, 3H), 2. 15(d,J＝4.7,1H),2.04(s,1H),1.94(d,J＝12.4,1H),1.84–1.24(m,22 H),1.21(s,6H),1.13–0.93(m,6H),0.90(d,J=6.4,3H),0.61(s,6H).

Step 2: In a 50 mL round-bottom flask at room temperature, 4-methylbenzenesulfonic acid-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methyl ester 33-1 (40 mg, 0.068 mmol) was dissolved in N,N-dimethylformamide (1 mL), sodium cyanide (20 mg, 0.41 mmol) was added at room temperature, and then stirred at 100 ° C for 16 hours, and monitored by TLC (petroleum ether: ethyl acetate = 3:1). The reaction was continued for 2-3 hr. After the reaction was completed, 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=3:1) to obtain a white solid [(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]acetonitrile 33 (15.71 mg, purity: 77.49%, yield: 40.39%).

Compound 33: ¹ H NMR (400 MHz, CDCl3) δ = 4.97 (s, 1H), 4.56 (s, 1H), 3.92 (dd, J = 11.4, 5.2, 1H), 1.95–1.88 (m, 2H), 1.81–1.24 (m, 23H), 1.14 (s, 6H), 1.08–0.93 (m, 6H), 0.85 (d, J = 6.5, 3H), 0.62 (s, 3H), 0.58 (s, 3H). ¹³ C NMR(101MHz, CDCl3)δ=153.33,102.36,73.37,71.13,56.28,56.23,54.49,49.90,44.42,42.62,40.08,38.62,37.27,36 .43,35.76,35.26,33.05,31.61,29.36,29.21,28.28,24.27,24.21,21.72,20.78,18.64,12.94,12.07.LC-MS: [M+H-2H ₂ O] ⁺ = 408.3

Embodiment 34

Compound 34 Preparation of 24,24-difluoro-5α-cholest-3β,4β,25-triol.

In the first step, (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluorohexanoic acid ethyl ester 24-3 (300 mg, 0.606 mmol, 1 eq) was dissolved in chloroform (10 mL), and selenium dioxide (201.8 mg, 1.82 mmol, 3 eq) and N-methylmorpholine (153.2 mg, 1.52 mmol, 2.5 eq) were added to the solution, and the mixture was stirred at room temperature for 20 hours. After the reaction was completed, 30 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL*3). The obtained organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product. The crude product was purified by silica gel column (petroleum ether: ethyl acetate = 10:1) to obtain (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluorohexanoic acid ethyl ester 34-1 (155 mg, 0.304 mmol, yield: 50%). ¹ H NMR (400 MHz, CDCl ₃ )δ5.70(d,J＝3.4Hz,1H),4.72(dt,J＝12.1,4.0Hz,1H),4.36–4.30(m,2H),4.25(d,J＝2.7Hz,1H),2 .10(s,3H),2.04–1.92(m,3H),1.89–1.83(m,2H),1.72–1.64(m,2H),1.57(dd,J＝10.2,4.9Hz,4H), 1.49–1.43(m,3H),1.36(d,J＝7.1Hz,3H),1.27(dd,J＝9.3,2.5Hz,2H),1.22(s,3H),1.17–1.09(m,3 H),1.07–0.99(m,2H),0.94(d,J＝6.5Hz,3H),0.87(dd,J＝19.1,8.6Hz,1H),0.70(d,J＝12.6Hz,3H).

Step 2 (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluorohexanoic acid ethyl ester 34-1 (155 mg, 0.304 mmol, 1 eq) was dissolved in ethyl acetate (15 mL), and then added thereto. Platinum dioxide (50%) (75 mg) was added, stirred at room temperature for 12 hours, filtered after the reaction was completed, and then the organic solvent was dried to obtain the product (5R)-5-[(1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-7-acetoxy-6-hydroxy-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluorohexanoic acid ethyl ester 34-2 (70 mg, 0.137 mmol, yield: 46%) as an oily compound. ¹ H NMR (400 MHz, cdcl ₃ )δ4.69–4.61(m,1H),4.25(q,J＝7.1Hz,2H),3.76(s,1H),2.02(s,3H),1.89–1.76(m,3 H),1.72–1.66(m,3H),1.59(dd,J＝12.6,3.7Hz,1H),1.46(dd,J＝32.8,19.9Hz,5H),1. 28(s,3H),1.16(dd,J＝22.6,9.6Hz,3H),1.08–1.01(m,3H),0.98(s,3H),0.96–0.87(m ,2H),0.85(d,J＝6.6Hz,3H),0.81(d,J＝4.7Hz,1H),0.58(s,3H),0.54(d,J＝3.0Hz,1H).

In the third step, compound (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2,2-difluorohexanoic acid ethyl ester 34-2 (70 mg, 0.137 mmol, 1 eq) was dissolved in tetrahydrofuran (5 mL), and methylmagnesium bromide (2.4 M, 0.6 mL, 10 eq) was slowly added dropwise after replacing nitrogen. The mixture was stirred at room temperature for 3 hours and monitored by TLC (petroleum ether:ethyl acetate = 1:1). The reaction was completed. After the reaction was completed, water was added to quench the reaction mixture, followed by extraction with ethyl acetate (50 mL). The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product, which was then separated and purified on silica gel (petroleum ether: ethyl acetate = 1:1) to obtain (1R, 3aS, 3bR, 7S, 9aS, 9bS, 11aR)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol 34 (11 mg, 0.024 mmol, yield: 18%).

Compound 34: ¹ H NMR (400 MHz, CDCl ₃ ) δ3.67 (s, 1H), 3.54–3.44 (m, 1H), 1.94–1.85 (m, 2H), 1.79–1.52 (m, 16H), 1.43–1.30 (m, 5H), 1.23 (s, 6H), 1.07–0.98 (m, 4H), 0.95 (s, 3H), 0.91 (d, J＝6.1 Hz, 2H), 0.85 (d, J＝6.6 Hz, 3H), 0.81 (s, 1H), 0.59 (s, 3H), 0.56–0.49 (m, 1H). ¹³ C NMR (101 MHz, CDCl ₃ ) δ74.84, 73.69,72.30,64.38,59.40,59.37,58.07,56.53,55.82,48.83,42.65,42.63,39.88,36.88, 35.50,35.40,35.38,34.55,28.09,25.86,24.18,23.60,20.59,18.78,18.48,14.67,12.09.

Example 41 & 65

Compound 41 (1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-4,4-difluoro-9a, 11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl] hexadecahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol

Preparation of compound 65 (1R, 3aR, 5aR, 6R, 7S, 9aR, 9bR, 11aR)-4-fluoro-9a, 11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl]-2,3,3a, 5,5a, 6,7,8,9,9a,9b, 10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol.

Step 1: (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-difluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanal I and (5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12bR)-11-fluoro-2,2,5a,7a-tetramethyl A mixture of 4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-tetradecahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-8-yl]hexanal II (70 mg, 0.150 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL). After complete dissolution, cesium fluoride (11.39 mg, 0.075 mmol, 0.5 eq) and (trifluoromethyl)trimethylsilane (106.7 mg, 0.750 mmol, 5 eq) were added to the reaction system in sequence. After the addition was completed, the atmosphere was replaced with nitrogen three times and stirred at room temperature for 1 hour. After the reaction was complete as monitored by TLC (petroleum ether: ethyl acetate = 5:1), tetrabutylammonium fluoride (1 mL, 1 mmol, 1 mol/L) was added, and nitrogen was replaced three times after the addition was completed, and the mixture was stirred at room temperature for 1 hour. After the reaction was complete as monitored by TLC (petroleum ether: ethyl acetate = 5:1), the mixture was quenched with water (10 mL), and the reaction solution was washed three times with water (~10 mL), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was separated and purified by flash chromatography (petroleum ether: ethyl acetate = 95:5 to 85:15) to obtain a white solid (6R)-6-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-difluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]-1,1,1-trifluoroheptyl]- -2-ol 41-1 and (6R)-1,1,1-trifluoro-6-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-fluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-tetradecahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-8-yl]heptan-2-ol 65-1 mixture (50 mg, 0.089 mmol, purity 80.0%, yield 59.62% calculated based on 41-1).

Compound 41-1: ¹ HNMR (400 MHz, CDCl ₃ )δ3.94 (dt, J＝8.3, 6.2 Hz, 2H), 2.11–1.95 (m, 1H), 1.93–1.85 (m, 2H), 1.83–1.71 (m, 4H), 1.66–1.50 (m, 6H), 1.49–1.43 (m, 5H), 1.41–1.27 (m, 5H), 1.23 (d, J＝3.4 Hz, 3H) ,1.19(dd,J＝9.0,5.3Hz,2H),1.05(dd,J＝10.5,7.0Hz,2H),1.01(s,3H),0.98–0.90 (m,1H),0.87(t,J＝5.8Hz,3H),0.82(dd,J＝13.7,3.1Hz,1H),0.60(d,J＝12.1Hz,3H).

Step 2: Resuspend the raw materials (6R)-6-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-difluoro-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]-1,1,1-trifluoroheptan-2-ol 41-1 and (6R)-1,1,1-trifluoro-6-[(3aS,5a The mixture of (50 mg, 0.093 mmol, 1.0 eq) and (50 mg, 0.093 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL), and hydrochloric acid (1 mL, 3 mol/L) was slowly added dropwise. The atmosphere was replaced with nitrogen three times and stirred at room temperature for 1 hr. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1: 1). The reaction solution was washed with saturated sodium bicarbonate solution (3 × 5 mL). The combined organic phases were washed with saturated brine (10 mL), then dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated and purified by flash chromatography (petroleum ether: ethyl acetate = 90: 10 to 80: 20), (1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-4,4-difluoro-9a, 11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl] hexahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol 41 and (1R, 3aR, 5aR, 6 A mixture of 65 (40 mg, yield 69.16% based on 41) of a mixture of (R,7S,9aR,9bR,11aR)-4-fluoro-9a,11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl]-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol was directly used for the next step.

Step 3: Compounds (1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4,4-difluoro-9a,11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl]hexahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol 41 and (1R,3aR,5aR,6R,7S,9aR,9bS,11aR)-4-fluoro-9a,11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl]-2,3,3a,5,5 A mixture of a, 6, 7, 8, 9, 9a, 9b, 10, 11, 11a-tetrahydro-1H-cyclopenta[1, 2-a]phenanthrene-6, 7-diol (23 mg, 0.046 mmol, 1.0 eq) was added to a reaction bottle containing dichloromethane (2 mL), and triethylamine (0.045 mL, 0.324 mmol, 7.0 eq) and benzoyl chloride (0.027 mL, 0.232 mmol, 5.0 eq) were added, and then stirred at room temperature (20°C) for 18 hr; the reaction was monitored by TLC (dichloromethane: methanol = 20: 1). Water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL*2). The organic phases were combined, washed with water (20 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45°C) to obtain a crude product, which was separated and purified by prep-TLC (dichloromethane: methanol = 20:1) and SFC separation (instrument: Waters Acquity UPCC; chromatographic column: Daicel CHIRALPAK ID_3, 3.0*150 mm, 3 um; mobile phase: A/B: CO ₂ /MeOH (0.1% DEA) = 75/25; flow rate: 2.0 ml/min; column temperature: 37 degrees) to give the product 41-2-benzoic acid-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4,4-difluoro-6-hydroxy-9a,11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl]hexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (20 mg, 0.030 mmol, 64.70%, retention time t _R = 1.096 min) as a white solid. Product 65-2: Benzoic acid-(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-4-fluoro-6-hydroxy-9a,11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl]-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (10 mg, 0.010 mmol, 22.38%, retention time t _R =1.280 min) was a white solid.

Compound 41-2: ¹ H NMR (400 MHz, CDCl3) δ8.08 (d, J = 7.6 Hz, 2H), 7.62 (t, J = 7.4 Hz, 1H), 7.48 (t, J = 7.7 Hz, 2H), 5.61–5.48 (m, 1H), 3.73 (s, 1H), 3.59 (d, J = 10.7 Hz, 1H), 2.28–2.13 (m, 1H), 2.12–2.07 (m, 1H), 1.94 (m, 1H), 1.88 (m, 1H), 1.83 ( m,2H),1.80(m,3H),1.75–1.69(m,2H),1.69–1.63(m,1H),1.56(m,1H),1.46(m,3H),1.40–1.34(m,3H),1.33 –1.29(m,2H),1.21(m,1H),1.13–1.06(m,2H),1.05(s,3H),0.98(m,2H),0.87(d,J＝6.4Hz,3H),0.64(s,3H). ¹⁹ F NMR (377MHz, CDCl3) δ-76.92,-77.02,-88.65,-89.28,-110.63,-111.26.

Compound 65-2: ¹⁹ F NMR (377 MHz, CDCl3) δ -76.91, -77.04, -109.74.

Step 4: Compound benzoic acid-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4,4-difluoro-6-hydroxy-9a,11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl]hexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 41-2 (20 mg, 0.033 mmol, 1.0 eq) was added to a reaction bottle containing methanol (0.5 mL) and tetrahydrofuran (1 mL), and a solution of lithium hydroxide (13.97 mg, 0.333 mmol) in water (0.5 mL) was added, and the temperature was raised to 40°C and stirred for 2 hr; TLC (petroleum ether:ethyl acetate = 1:1) monitoring of the reaction showed that the starting material disappeared and there was a polar change. A large main point was generated; water (20 mL) was added to the reaction solution, extracted with ethyl acetate (20 mL*3), the organic phases were combined, washed with water (20 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was decompressed and dried (water pump, 45°C) to obtain a crude product, which was separated and purified by column chromatography (petroleum ether: ethyl acetate = 1:0-2:1) to obtain the product (1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-4,4-difluoro-9a, 11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl] hexahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol 41 (16.40 mg, 0.033 mmol, 99.21%) as a white solid.

Compound 41: ¹ H NMR (400 MHz, CDCl ₃ )δ3.91(m,1H),3.74(s,1H),3.60(dt,J＝11.3,4.1Hz,1H),2.21(m,1H),1.98( m,2H),1.86(m,3H),1.81–1.75(m,2H),1.74–1.71(m,1H),1.70–1.62(m,3H),1 .57(m,2H),1.46(m,3H),1.41–1.37(m,2H),1.35–1.28(m,2H),1.25(m,1H),1. 11(m,2H),1.06(s,3H),1.04–0.95(m,2H),0.93(d,J＝6.5Hz,3H),0.67(s,3H). ¹⁹ F NMR (377MHz, CDCl ₃ ) δ -80.03, -88.64, -89.27, -110.64, -111.26. ¹⁹ F NMR (377MHz, CDCl 3 ) δ -76.92, -77.02, -88.65, -89.28, -110.63, -111.26.

Compound benzoic acid-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4-fluoro-6-hydroxy-9a,11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl]hexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 65-2 (10 mg, 0.017 mmol, 1.0 eq) was added to a reaction bottle containing methanol (0.5 mL) and tetrahydrofuran (1 mL), and a solution of lithium hydroxide (7.20 mg, 0.172 mmol) in water (0.5 mL) was added, and the temperature was raised to 40°C and stirred for 2 hr; the reaction was monitored by TLC (petroleum ether:ethyl acetate = 1:1); water (20 mL) was added to the reaction solution, and ethyl acetate (2 The organic phases were combined, washed with water (20 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45°C) to give a crude product, which was separated and purified by column chromatography (petroleum ether:ethyl acetate=1:0-2:1) to give the product (1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-4-fluoro-9a,11a-dimethyl-1-[(2R)-7,7,7-trifluoro-6-hydroxyhept-2-yl]-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol 65 (1.89 mg, 0.004 mmol, 21.44%) as a white solid.

Compound 65: ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.91 (s, 1H), 3.85 (s, 1H), 3.59 (d, J = 11.0 Hz, 1H), 2.66 (t, J = 12.5 Hz, 1H), 2.07–1.98 (m, 2H), 1.94 (d, J = 16.5 Hz, 2H), 1.88–1.82 (m, 3H), 1.80–1.71 (m, 3H), 1.68 (m, 1H), 1.63–1.59 (m, 3H), 1.55 (m, 2H), 1.48 (m, 2H), 1.45 (m, 3H), 1.13 (m, 4H), 1.05 (s, 3H), 0.94 (d, J = 6.5 Hz, 3H), 0.65 (s, 3H). ¹⁹ F NMR (376MHz, CDCl ₃ ) δ -80.03, -109.71. ¹⁹ F NMR (377MHz, CDCl ₃ ) δ -76.91, -77.04, -109.74.

Embodiment 53

Preparation of Compound 53 4-(1-hydroxyethyl)-5α-cholest-3β,25-diol

Step 1: In a 50 mL round-bottom flask at room temperature, 3β-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-25-hydroxy-5α-cholestane-4-carboxaldehyde 23-3 (23 mg, 0.042 mmol) was dissolved in anhydrous tetrahydrofuran (1.5 mL). The reaction was cooled to 0°C under nitrogen protection, and 3 mol/L methylmagnesium bromide tetrahydrofuran solution (0.07 mL, 0.021 mmol) was added dropwise. Then, the mixture was stirred at room temperature for 2 hours. TLC (petroleum ether: ethyl acetate = 4:1) was performed. Monitor the reaction. After the reaction is completed, the system is cooled to 0°C, quenched with saturated aqueous ammonium chloride solution (20 mL), extracted with ethyl acetate (10 mL×3), and the organic phase is washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain a white solid 3β-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-4-(1-hydroxyethyl)-5α-cholest-25-ol 53-1 (17 mg, purity: 90%, yield: 64.64%). ¹ HNMR(400MHz, CDCl3)δ=4.20-4.11(m,1H),3.85–3.70(m,1H),2.04–1.65(m,7H),1.60–1.46(m,6H) ,1.39–1.29(m,5H),1.21(s,6H),0.94(s,3H),0.91(dd,J＝6.9,4.1,12H),0.64(s,3H),0.06(s,6H).

Step 2: In a 50 mL round-bottom flask at room temperature, 3β-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-4-(1-hydroxyethyl)-5α-cholest-25-ol 53-1 (17 mg, 0.03 mmol) was dissolved in a 1 mol tetrabutylammonium fluoride tetrahydrofuran solution (0.5 mL) and 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 system was cooled to 0°C, quenched with saturated aqueous ammonium chloride solution (10 mL), extracted with ethyl acetate (5 mL × 3), and the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1: 1) to obtain a white solid 4-(1-hydroxyethyl)-5α-cholest-3β,25-diol 53 (5.09 mg, purity: 95.12%, yield: 35.73%).

Compound 53: ¹ H NMR (400 MHz, CDCl3) δ＝4.19–4.08 (m, 1H), 3.83–3.71 (m, 1H), 1.96–1.65 (m, 7H), 1.56–1.47 (m, 5H), 1.42–1.17 (m, 18H), 1.14 (s, 6H), 1.07–0.92 (m, 5H), 0.88 (s, 3H), 0.84 (d, J＝6.5, 3H), 0.57 (s, 3H). ¹³ C NMR(101MHz, CDCl3)δ=104.61,103.05,101.06,71.13,69.23,56.48,56.3 7,56.18,44.43,42.48,39.95,38.55,36.44,35.88,35.75,35.73,33.64,3 3.59,31.45,30.97,30.36,29.37,29.21,29.04,28.26,26.29,25.61,24.8 0,24.27,21.07,20.99,20.92,20.78,18.64,14.96,12.01.LC-MS: [M+H-2H ₂ O] ⁺ =413.55

Embodiment 55

Compound 55 Preparation of 4-bromo-5α-cholest-3β,25-diol

First step In a 100 mL round-bottom flask, (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-8 (500 mg, 1.09 mmol) was dissolved in ethyl acetate (30 mL), 10% palladium carbon (250 mg) and acetic acid (0.3 mL) were added at room temperature, and the mixture was stirred under hydrogen atmosphere at room temperature for 5 hours. The reaction was monitored by TLC (ethyl acetate/petroleum ether=8:1). After the reaction, the reaction solution was diluted with ethyl acetate (60 mL), filtered through diatomaceous earth, the organic phase was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8: 1) to obtain a white solid (5R)-5-[(1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-7-acetoxy-6-hydroxy-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 55-1 (390 mg, purity: 90%, yield: 62.72%). ¹ H NMR (400MHz, CDCl3) δ4.75–4.69(m,1H),3.83(s,1H),3.66(s,3H),2.35–2.20(m,2H ),2.08(s,3H),2.03–1.05(m,27H),1.05(s,3H),0.92(d,J=6.5Hz,3H),0.64(s,3H).

Step 2: In a 50 mL round-bottom flask at room temperature, (5R)-5-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 55-1 (250 mg, 0.54 mmol) was dissolved in 1,2-dichloroethane (10 mL), and a solution of triphenylphosphine (142 mg, 0.54 mmol) in 1,2-dichloroethane (0.6 mL) was added at 0°C. The reaction solution was stirred at room temperature for 10 minutes, and a solution of carbon tetrabromide (180 mg, 0.54 mmol) in 1,2-dichloroethane (0.9 mL) was added to the reaction solution at 0°C. The reaction solution was then warmed to room temperature and stirred at 50°C for 16 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 5: 1). After the reaction was completed, the mixture was quenched with water (30 mL), extracted with ethyl acetate (20 mL × 3), washed with saturated brine (40 mL) and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain a white solid (5R)-5-[(1R, 3aS, 3bS, 5aR, 7S, 9aR, 9bS, 11aR)-7-acetoxy-6-bromo-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 55-2 (150 mg, purity: 90%, yield: 47.57%). ¹ H NMR (400MHz, CDCl3) δ = 4.97 (s, 1H), 4.32 (s, 1H), 3.67 (s, 3H), 2.34–2.18 (m, 3H), 2.05 (s, 3H), 1.97 (dd ,J＝8.0,4.4,2H),1.89–1.36(m,19H),1.18–1.03(m,5H),0.98(s,3H),0.92(d,J＝6.5,3H),0.64(s,3H).

Step 3: The reactant (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-acetoxy-6-bromo-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 55-2 (150 mg, 0.337 mmol, 1 eq) was dissolved in tetrahydrofuran (10 mL), and the reaction system was replaced with a nitrogen atmosphere. After the reaction system was cooled to 0°C, methyl magnesium chloride (1.124 mL, 3.373 mmol, 1 mol/L) was slowly added and stirred at room temperature for 1 h. TLC (petroleum ether: ethyl acetate = 5:1) was used to monitor the reaction. 20 mL of water was added to the reaction system, extracted with ethyl acetate (25 mL×3), the organic phase was collected, dried with anhydrous sodium sulfate, and the organic phase was vacuum dried to obtain a crude product. The crude product was dissolved in ethyl acetate and purified by silica gel column chromatography (petroleum ether:ethyl acetate=88:12) to obtain 4-bromo-5α-cholest-3β,25-diol 55 (16 mg, purity: 90.51%, yield: 10.66%).

Compound 55: ¹ H NMR(400MHz,cdcl3)δ4.37(d,J＝2.6Hz,1H),3.87(s,1H),2.41–2.30(m,1H),2 .00–1.94(m,1H),1.87–1.73(m,4H),1.64(dd,J＝12.9,4.0Hz,1H),1.54(dd,J ＝9.0,3.4Hz,6H),1.49–1.34(m,9H),1.22(s,6H),1.17–1.04(m,4H),1.02(s, 3H),0.99–0.95(m,1H),0.92(d,J＝6.5Hz,3H),0.87–0.72(m,2H),0.65(s,3H).

Embodiment 56

Compound 56 Preparation of 4β-[(ethoxymethyl)oxy]-3-methyl-5α-cholest-3,25-diol.

Step 1: Dissolve (1R,7S,9aR,11aR)-6-hydroxy-1-[(2R)-6-methoxy-6-oxyylidenehexan-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl acetate 55-1 (100 mg, 0.216 mmol) in N,N-diisopropylethylamine (5 mL), add [(chloromethyl)oxy]ethane (204.21 mg, 2.16 mmol), react at 130 ° C for 1.5 h, monitor by TLC (petroleum ether: ethyl acetate = 5:1) and NMR The raw material was no longer left. The product was washed with ethyl acetate and water, dried over anhydrous sodium sulfate, and concentrated. The product was purified by column chromatography (petroleum ether:ethyl acetate = 100:1 to 20:1) to obtain acetic acid-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-[(ethoxymethyl)oxy]-1-[(2R)-6-methoxy-6-oxyylidenehexan-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 56-1 (60 mg, 0.106 mmol, 49.04%) as a white solid. ¹ HNMR(399MHz,Chloroform-d)δ4.71–4.60(m,3H),3.70(d,J＝8.7Hz,1H),3.65( s,3H),2.23(tq,J＝16.8,9.6,9.1Hz,2H),2.03(s,3H),1.91(t,J＝12.5Hz,2H),1 .53(s,3H),1.39–1.30(m,4H),1.26(d,J＝14.6Hz,2H),1.17(d,J＝7.3Hz,3H),1 .13(s,1H),1.10–1.01(m,4H),0.99(s,4H),0.90(d,J=6.6Hz,3H),0.62(s,3H).

Step 2: Dissolve (1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-6-[(ethoxymethyl)oxy]-1-[(2R)-6-methoxy-6-oxyylidenehexan-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl acetate 56-1 (60 mg, 0.115 mmol) in methanol (5 mL), add potassium carbonate (158.70 mg, 1.150 mmol) at room temperature, stir at room temperature for 5 h, and TLC (petroleum ether: acetic acid) was performed. After the reaction, water and ethyl acetate were added for extraction, the organic phase was separated and the crude product was dried, and purified by column chromatography (petroleum ether:ethyl acetate=60:1-3:1) to obtain (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-[(ethoxymethyl)oxy]-7-hydroxy-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 56-2 (40 mg, 0.079 mmol, 58.89%) as a white solid. ¹ H NMR(399MHz,Chloroform-d)δ4.80(d,J＝6.7Hz,1H),4.53(d,J＝6.7Hz,1H),3.76(dq,J＝9.5,7.1Hz,1H),3.65( s,3H),3.60–3.49(m,2H),3.39(s,1H),2.31–2.19(m,2H),1.96–1.90(m,1H),1.68(s,2H),1.57–1.44(m,2H), 1.42–1.32(m,3H),1.31–1.25(m,2H),1.22(t,J＝7.1Hz,5H),1.14(d,J＝9.5Hz,1H),1 .11–1.02(m,4H),1.01–0.95(m,1H),0.93(s,3H),0.90(d,J=6.6Hz,3H),0.62(s,3H).

Step 3: Dissolve (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-[(ethoxymethyl)oxy]-7-hydroxy-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 56-2 (20 mg, 0.042 mmol) in dichloromethane (5 mL), add N-methylmorpholine oxide (7.34 mg, 0.063 mmol) and tetrapropylammonium perruthenate (1.47 mg, 0.004 mmol) at room temperature, and heat for 2 h. The mixture was stirred for 1 h and monitored by TLC (petroleum ether:ethyl acetate=5:1). After the reaction, the mixture was filtered through diatomaceous earth, the filtrate was concentrated, and the mixture was purified by column chromatography (petroleum ether:ethyl acetate=60:1-3:1) to obtain (5R)-5-[(1R,3aS,3bS,5aR,6R,9aR,9bS,11aR)-6-[(ethoxymethyl)oxy]-9a,11a-dimethyl-7-oxyylidenehexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 56-3 (15 mg, 0.029 mmol, 69.08%) as a white solid. ¹ H NMR(399MHz,Chloroform-d)δ4.65–4.52(m,2H),3.65(s,3H),3.60–3.48(m,3H),2.29–2.13(m,3H),1.37(d,J＝19.8Hz,7H),1 .24(s,2H),1.18(d,J＝4.7Hz,3H),1.15(d,J＝7.3Hz,3H),1.11–1.03(m,3H),0.96(s,1H),0.90(d,J＝6.6Hz,3H),0.65(s,4H).

Step 4: Dissolve (5R)-5-[(1R,3aS,3bS,5aR,6R,9aR,9bS,11aR)-6-[(ethoxymethyl)oxy]-9a,11a-dimethyl-7-oxoylidenehexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 56-3 (20 mg, 0.042 mmol) in tetrahydrofuran (3 mL), and add methyl lithium (9.22 m g, 0.42 mmol), stirred at room temperature for 2 h, monitored by TLC (petroleum ether: ethyl acetate = 2:1); after the reaction, water and ethyl acetate were added for extraction, the organic phase was separated and the crude product was dried, and purified by column chromatography (petroleum ether: ethyl acetate = 50:1 to 1:1) to obtain 4β-[(ethoxymethyl)oxy]-3-methyl-5α-cholest-3,25-diol 56 (10 mg, 0.019 mmol, 45.90%) as a white solid.

Compound 56: ¹ H NMR (400 MHz, Chloroform-d) δ 4.78 (d, J = 6.6 Hz, 1H), 4.60 (d, J = 6.6 Hz, 1H), 3.73–3.57 (m, 2H), 3.15 (s, 1H), 1.94 (d, J = 12.6 Hz, 1H), 1.74 (t, J = 14.7 Hz, 3H), 1.65 (m, 2H), 1.53-1.61 (m, 5H), 1.43 (d ,J＝15.2Hz,3H),1.36(d,J＝10.6Hz,3H),1.29(d,J＝22.8Hz,3H),1.23(s,3H),1.22–1.16(m,12H), 1.12–1.05(m,2H),1.01(dd,J＝19.5,8.8Hz,3H),0.94(s,3H),0.89(d,J＝6.5Hz,3H),0.62(s,3H). ¹³ CNMR(101MHz,Chloroform-d)δ98.16,90.81,71.10,70.96,64.32,56.48,56.12,47.42,44.37,42.57,39.86,36. 13,35.73,35.47,32.89,32.31,29.18,28.23,26.75,24.55,24.18,20.75,18.60,15.09,12.02.ELSD-MS:[Ms+NH ₄ ] ⁺ =491.42; [Ms-H ₂ O-OH] ⁺ =493.42. HPLC: 94.97% (CAD).

Embodiment 59

Compound 59 Preparation of 3-methyl-5α-cholest-4β,3,25-triol.

4β-[(Ethoxymethyl)oxy]-3-methyl-5α-cholestane-3,25-diol 56 (50 mg, 0.101 mmol) was dissolved in tetrahydrofuran (3 mL), p-toluenesulfonic acid (174.08 mg, 1.014 mmol) was added at room temperature, and the mixture was heated at 40°C with stirring for 1 h and monitored by TLC (petroleum ether:ethyl acetate = 1:1). After the reaction, water and ethyl acetate were added for extraction, the organic phase was separated, the crude product was dried over anhydrous sodium sulfate, and purified by column chromatography (petroleum ether:ethyl acetate = 50:1 to 1:1) to obtain 3-methyl-5α-cholestane-4β,3,25-triol 59 (20 mg, 0.044 mmol, 45.35%) as a white solid.

Compound 59: ¹ H NMR(399MHz,Chloroform-d)δ3.32(s,1H),2.18(s,1H),1.94(d,J＝12.8Hz,1H),1.63(s,1H),1.76(s,3H), 1.53(s,4H),1.42(d,J＝13.9Hz,3 H),1.39–1.32(m,5H),1.31(s,3H),1.23(s,3H),1.19(s,6H),1.04(dd,J＝11.3,6.8Hz,4H),1.00(s ,3H),0.89(d,J＝6.5Hz,3H),0.62(s,3H). ^13C NMR (100MHz, Chloroform-d) δ72.14,71.09,56.54,56.13,55.50,46.96,44.38,42.58,39.86,36.39,36.23,35.78,35. 72,35.45,32.41,31.57,29.34,29.17,28.23,26.06,24.80,24.18,20.74,20.54,18.61,14.64,12.05.ELSD-MS:[Ms+NH ₄ ] ⁺ =433.38; [Ms-H ₂ O-OH] ⁺ =435.38. HPLC: 93.78% (CAD).

Embodiment 60

Compound 60 Preparation of 24-fluoro-5α-cholest-3β,4β,25-triol.

Step 1: Weigh acetic acid-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-hydroxy-1-[(2R)-6-methoxy-6-oxyhexan-2-yl]-9a,11a-dimethylhexadecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester IV-1 (300 mg, 0.65 mol, 1.0 eq.) and dissolve it in anhydrous methanol (5 mL), add potassium carbonate (897 mg, 6.5 mmol), then raise the temperature to 25 °C and stir for 16 hours, monitor by TLC (petroleum ether: ethyl acetate = 2:1), and then the reaction is completed. The reaction solution was filtered and concentrated, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate = 0-50%) to give (5R)-5-[(1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-6,7-dihydroxy-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester IV-2 (210 mg, 0.5 mmol, 76.9%) as a white solid. 1H NMR(399MHz,Chloroform-d)δ3.71(d,J＝3.1Hz,1H),3.65(s,3H),3.53(dt,J＝11.3,4.4Hz,1H),2.33–2.18(m,2H),1.97–1.84(m,2H),1.83–1. 60(m,8H),1.42–1.28(m,5H),1.28–1.15(m,3H),1.11–1.01(m,4H),1.00 (s,3H),0.90(d,J＝6.5Hz,4H),0.62(s,3H),0.57(td,J＝11.4,4.2Hz,1H).

Step 2: Weigh (5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-dihydroxy-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester IV-2 (250 mg, 0.59 mmol) and dissolve it in acetone (20 mL). Add 4A molecular sieves (250 mg) and p-toluenesulfonic acid (203 mg, 1.18 mmol) and stir at room temperature. Monitor by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction was completed, the insoluble matter was removed by filtration, and the filtrate was concentrated and purified by column chromatography (petroleum ether: ethyl acetate = 0-30%) to obtain (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-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][ ^1,3 ]dioxolan-8-yl]hexanoic acid methyl ester IV-3 (220 mg, 0.48 mmol, 81.3%) as a white solid. NMR(399MHz,Chloroform-d)δ3.97(dt,J＝8.8,5.7Hz,2H),3.65(s,3H),2.25(ddd,J＝10.5,8.5,6.7Hz,2 H),1.97–1.90(m,1H),1.76(td,J＝11.5,6.6Hz,3H),1.71–1.63(m,3H),1.58–1.52(m,1H),1.49(s,4H), 1.47–1.32(m,6H),1.28(s,3H),1.23(ddd,J＝14.1,8.9,5.6Hz,3H),1.08(td,J＝10.7,9.2,3.7Hz,3H),1 .02(s,3H),0.98–0.93(m,1H),0.90(d,J＝6.4Hz,3H),0.86–0.80(m,2H),0.64(s,3H),0.61–0.54(m,1H).

Step 3: In a three-necked flask, (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester IV-3 (370 mg, 0.803 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (7 mL), and trimethylsilyl chloride (0.440 mL, 2.409 mmol, 3.0 eq.) was added. Replace the three-necked flask with nitrogen atmosphere, then cool to -78°C and add lithium diisopropylamide (172.08 mg, 1.606 mmol, 2.0 eq.). The reaction solution is then heated to 0°C and stirred for 15 minutes. Add a tetrahydrofuran (1 mL) solution of N-fluorobisbenzenesulfonamide (506.50 mg, 1.606 mmol, 2.0 eq.). The reaction solution is heated to room temperature and stirred for 1 hour. Monitor by TLC (petroleum ether: ethyl acetate = 10:1). When the raw materials are completely reacted, add water to the reaction solution to quench. The obtained mixed solution was extracted three times with ethyl acetate, and the combined organic phase was washed once with brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate/petroleum ether = 0-20%) to obtain (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro- ^3aH -cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]-2-fluorohexanoic acid methyl ester as a white solid 60-1 (200 mg, 0.397 mmol, 49.42%). NMR(399MHz,Chloroform-d)δ4.14(ddd,J＝48.5,18.2,10.2Hz,1H),3.71(d,J＝3.0Hz,1H),3.53(dt,J＝11.4,4.3Hz,1H) ,1.94(dt,J＝12.5,3.5Hz,1H),1.82(dd,J＝9.0,5.3Hz,2H),1.76–1.71(m,3H),1.70–1.62(m,3H),1.59–1.46(m,2H),1.3 7(ddd,J＝19.3,12.0,4.8Hz,4H),1.30–1.21(m,3H),1.19(d,J＝5.0Hz,6H),1.13–1.02(m,4H),0.99(s,3H),0.94(dd,J＝8 .7,4.3Hz,2H),0.90(dd,J＝6.5,2.0Hz,3H),0.87–0.79(m,1H),0.63(s,3H),0.57(td,J＝11.4,4.2Hz,1H).LC-MS:[M-HF] ⁺ =458.45;

Step 4: Dissolve (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-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]-2-fluorohexanoic acid methyl ester 60-1 (100 mg, 0.209 mmol, 1 eq.) in tetrahydrofuran (5 mL). Cool to 0°C under nitrogen atmosphere, add methyl lithium (0.653 mL, 1.045 mmol, 5.0 eq.). Heat to room temperature and stir for 1 hour. TLC (petroleum ether: ethyl acetate = 10:1) was used for monitoring. After the reaction was completed, a small amount of water was added to quench the reaction, and the mixture was directly concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate/petroleum ether = 0-30%) to obtain (6R)-6-[(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-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]-3-fluoro-2-methylheptan-2-ol as a white solid 60-2 (75 mg, 0.149 mmol, 71.24%). ¹ H NMR(399MHz,Chloroform-d)δ4.14(ddd,J＝48.4,18.5,10.3Hz,1H),4.01–3.92(m,2H),1.94(d,J ＝12.2Hz,1H),1.79(s,2H),1.64(dd,J＝15.6,12.1Hz,4H),1.54(s,5H),1.49(s,3H),1.44(d,J＝14 .1Hz,6H),1.28(s,3H),1.24(d,J＝11.6Hz,3H),1.19(d,J＝4.9Hz,6H),1.07(d,J＝11.1Hz,4H),1.0 2(s,3H),0.97(s,1H),0.92–0.88(m,3H),0.87–0.79(m,2H),0.65(s,3H),0.58(t,J＝10.2Hz,1H).

Step 5: (6R)-6-[(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-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]-3-fluoro-2-methylheptan-2-ol 60-2 (75 mg, 0.157 mmol) was dissolved in tetrahydrofuran (5 mL), and hydrochloric acid (2 mL, 4.000 mmol, 25.5 eq.) was added. The mixture was stirred at room temperature for 1 hour. TLC (petroleum ether:ethyl acetate = 5:1) was used for monitoring. The reaction solution was diluted with water and extracted three times with ethyl acetate. The organic phases were combined, washed once with brine, dried over anhydrous sodium sulfate, concentrated to give a crude product, and purified by column chromatography (ethyl acetate/petroleum ether = 0-60%) to give 24-fluoro-5α-cholest-3β,4β,25-triol as a white solid 60 (40 mg, 0.087 mmol, 55.29%).

Compound 60: ¹ H NMR (399 MHz, Chloroform-d) δ 4.14 (ddd, J = 48.5, 18.2, 10.2 Hz, 1H), 3.71 (d, J = 3.0 Hz, 1H), 3.53 (dt, J = 11.4, 4.3 Hz, 1H), 1.94 (dt, J = 12.5, 3.5 Hz, 1H), 1.82 (dd, J = 9.0, 5.3 Hz, 2H), 1.76–1.71 (m, 3H), 1.70–1.62 (m, 3H), 1.59–1.46 (m, 2H), 1.37 (ddd ,J＝19.3,12.0,4.8Hz,4H),1.30–1.21(m,3H),1.19(d,J＝5.0Hz,6H),1.13–1.02(m,4H),0.99(s,3H),0.94(dd,J＝8.7,4.3H z, 2H), 0.90 (dd, J＝6.5, 2.0Hz, 3H), 0.87–0.79 (m, 1H), 0.63 (s, 3H), 0.57 (td, J＝11.4, 4.2Hz, 1H). LC-MS: [M+Na]+ = 461.45.

Embodiment 61

Preparation of compound 61,3,25-dihydroxy-3-methyl-5α-cholest-4-one.

3-Methyl-5α-cholest-4β,3,25-triol 59 (20 mg, 0.046 mmol) was dissolved in dichloromethane (3 mL), and N-methylmorpholine oxide (8.08 mg, 0.069 mmol) and tetrapropylammonium perruthenate (1.61 mg, 0.005 mmol) were added at room temperature. The mixture was stirred at room temperature for 1 h and monitored by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction, the mixture was filtered with diatomaceous earth, and the filtrate was concentrated and purified by column chromatography (petroleum ether: ethyl acetate = 50:1 to 1:1) to obtain 3,25-dihydroxy-3-methyl-5α-cholest-4-one 61 (10 mg, 0.020 mmol, 85.85%) as a white solid.

Compound 61: ¹ HNMR (399MHz, Chloroform-d) δ2.33–2.37 (m, 1H), 2.07–1.96 (m, 2H), 1.85–1.71 (m, 4H), 1.46–1.40 (m, 4H), 1.38 (s, 3H), 1.37–1.31 (m, 3H), 1.24 (d, J=12.6Hz, 5H), 1.19 (s, 6H) ,1.17–1.10(m,2H),1.10–1.04(m,2H),1.01(d,J＝8.9Hz,2H),0.97–0.93(m,1H),0.90( d,J＝6.5Hz,3H),0.80(dt,J＝13.1,8.9Hz,1H),0.69(s,3H),0.63(s,3H).ELSD-MS:[M+NH ₄ ] ⁺ =433.38; [MH ₂ O-OH] ⁺ =435.38.

Embodiment 66

Compound 66: Preparation of (1R,3aR,7S,9aS,9bR,11aR)-4-fluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol.

Step 1: Dissolve (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-7 (1 g, 2.25 mmol, 1 eq) in tetrahydrofuran (10 mL), add cobalt acetate (80 mg, 0.45 mmol, 0.2 eq), N-hydroxyphthalimide (0.15 g, 0.90 mmol, 0.4 eq), tert-butyl peroxide (1.01 g, 11.25 mmol, 5 eq) to the solution, and stir at room temperature for 20 hours. After the reaction was completed, 30 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL*3). The obtained organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product. The crude product was purified by silica gel column (petroleum ether/ethyl acetate = 10/1) to obtain (5R)-5-[(1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-7-acetoxy-9a, 11a-dimethyl-4-oxyylidene-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 66-1 (0.6 g, 1.31 mmol, yield: 58%). ¹ H NMR (400 MHz, CDCl ₃ )δ5.70(d,J＝1.5Hz,1H),4.80–4.64(m,1H),3.67(s,3H),2.48(dddd,J＝15.6,12.1,9.1,2.4 Hz,3H),2.25(ddd,J＝22.3,12.9,5.3Hz,3H),2.05(s,3H),2.01–1.95(m,2H),1.93–1.84(m, 1H),1.79–1.65(m,2H),1.65–1.46(m,5H),1.40(ddd,J＝11.6,8.9,4.3Hz,2H),1.28(ddd,J＝ 10.9, 10.5, 5.9Hz, 3H), 1.21 (s, 3H), 1.18–1.05 (m, 3H), 0.94 (d, J = 6.5Hz, 3H), 0.68 (s, 3H).

Step 2: Compound (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-4-oxyde-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 66-1 (0.6 g, 1.31 mmol, 1 eq) was dissolved in ethyl acetate (15 mL), and then Palladium carbon (10%) (120 mg) was added thereto, and the mixture was stirred at room temperature for 12 hours. After the reaction was completed, the mixture was filtered, and then the organic solvent was dried by spin drying to obtain the product (5R)-5-[(1R, 3aS, 3bR, 7S, 9aS, 9bS, 11aR)-7-acetoxy-9a, 11a-dimethyl-4-oxyylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 66-2 (450 mg, 0.977 mmol, yield: 75%) as an oily compound. ¹ H NMR (400 MHz, cdcl ₃ )δ5.36–5.24(m,1H),4.30(s,3H),3.00–2.81(m,5H),2.70–2.58(m,5H),2.55–2.4 7(m,2H),2.42(dt,J＝13.4,3.5Hz,1H),2.35–2.26(m,2H),2.22–2.10(m,6H),2.09– 1.99(m,3H),1.86(d,J＝12.4Hz,1H),1.76(s,1H),1.73(s,1H),1.73(s,3H),1.70(s ,1H),1.69(d,J＝3.3Hz,1H),1.67–1.60(m,1H),1.56(d,J＝6.5Hz,3H),1.28(s,3H).

Step 3: Compound (5R)-5-[(1R, 3aS, 3bR, 7S, 9aS, 9bS, 11aR)-7-acetoxy-9a, 11a-dimethyl-4-oxyylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 66-2 (150 mg, 0.326 mmol, 1.0 eq) and DAST (157 mg, 0.977 mmol, 3 eq) were dissolved in tetrahydrofuran (5 mL), stirred at 80 ° C for 3 hours, and detected by TLC. After the reaction was completed, water was added to destroy it, and then extracted with ethyl acetate (50 mL). The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried to obtain a crude product, which was then separated and purified by silica gel (petroleum The reaction mixture was stirred for 2 h (40 ℃ of drying) and the mixture was stirred for 30 min. The mixture was stirred for 40 min. The mixture was stirred for 20 min. The mixture was stirred for 30 min. The mixture was stirred for 40 min. The mixture was stirred for 20 min. The mixture was stirred for 30 min. The mixture was stirred for 20 min. The mixture was stirred for 30 min. The mixture was stirred for 40 min. The mixture was stirred for 30 min.

Compound 66-3': ¹ H NMR (400 MHz, CDCl ₃ ) δ 4.75–4.63 (m, 1H), 3.66 (s, 3H), 2.34–2.22 (m, 2H), 2.10–1.91 (m, 5H), 1.84 (ddd, J = 9.3, 7.6, 3.7 Hz, 3H), 1.78–1.65 (m, 5H), 1.57–1.49 (m, 3H), 1.45–1.22 (m, 7H), 1.09 (ddd, J = 28.8, 14.4, 9.4 Hz, 5H), 0.94 (t, J = 5.4 Hz, 3H), 0.85 (d, J = 11.0 Hz, 3H), 0.71–0.62 (m, 3H).

Step 4: (R)-5-((3S, 5R, 9R, 10S, 13R, 14R, 17R)-3-acetoxy-7-fluoro-10,13-dimethyl-2,3,4,5,6,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl)hexanoic acid methyl ester 66-3 and (R)-5-((3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R)-3 A mixture of methyl 7,7-difluoro-10,13-dimethylhexahydro-1H-cyclopentyl[a]phenanthrene-17-yl)hexanoate 66-3' (70 mg, 0.145 mmol, 1 eq) was dissolved in tetrahydrofuran (5 mL), and methylmagnesium bromide (2.4 M) (0.6 mL, 10 eq) was slowly added dropwise after replacing nitrogen. The mixture was stirred at room temperature for 3 hours and detected by TLC. After the reaction was completed, water was added to quench the reaction, and then ethyl acetate (50 mL) was used for extraction. The organic phase was taken, washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product, which was then separated and purified on silica gel (petroleum ether: ethyl acetate = 1:1) to obtain (3S, 5R, 9R, 10S, 13R, 14R, 17R)-7-fluoro-17-((R)-6-hydroxy-6-methylheptane-2-yl)-10,13-dimethyl-2,3,4,5,6,9,10,11,12,13,14,15,16 A mixture of 17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-ol 66-4 and (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R)-7,7-difluoro-17-((R)-6-hydroxy-6-methylheptane-2-yl)-10,13-dimethylhexahydro-1H-cyclopenta[a]phenanthrene-3-ol 66-4' (22 mg, 0.05 mmol, yield calculated based on 66-4': 34%) was directly used for the next step.

Step 5: A mixture of (3S, 5R, 9R, 10S, 13R, 14R, 17R)-7-fluoro-17-((R)-6-hydroxy-6-methylheptane-2-yl)-10,13-dimethyl-2,3,4,5,6,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopentadien[a]phenanthrene-3-ol 66-4 and (3S, 5R, 8R, 9S, 10S, 13R, 14S, 17R)-7,7-difluoro-17-((R)-6-hydroxy-6-methylheptane-2-yl)-10,13-dimethylhexahydro-1H-cyclopenta[a]phenanthrene-3-ol 66-4' (1.3 g, 2.95 mmol, 1 eq) was added. Dissolved in dichloromethane (25 mL), slowly added triethylamine (1.19 g, 11.80 mmol, 4 eq), added DMAP (0.29 g, 2.36 mmol, 0.8 eq), slowly added benzoyl chloride (1.04 g, 7.38 mmol, 2.5 eq) at room temperature, stirred at room temperature for 1 hour, detected by TLC, after the reaction was completed, water was added to quench, and then extracted with ethyl acetate (50 mL), the organic phase was washed twice with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered, and dried to give a crude product, which was then separated and purified on silica gel (petroleum ether: ethyl acetate = 1:1) to give a mixture of 66-5 and 66-5' (1.5 g, 2.82 mmol, yield: 95%). After SFC separation (instrument: Waters Acquity UPCC; column: Daicel CHIRALPAK OZ3, 3*150 mm, 3 um; mobile phase: A/B: CO2/MeOH (0.1% DEA) = 70/30; flow rate: 2.0 ml/min; column temperature: 37 degrees), (6R)-6-[(1R, 3aS, 3bR, 7S, 9aS, 9bS, 11aR)-7-(benzyloxy)-4,4-difluoro-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylheptan-2-ol 66-5' (1 g, retention time t _R =1.178 min) and (1R,3aR,7S,9aS,9bR,11aR)-4-fluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol 66-5 (200 mg, retention time t _R =1.601 min).

Compound 66-5: ¹ H NMR (400 MHz, CDCl ₃ ) δ8.07–7.99 (m, 2H), 7.55 (t, J=7.4 Hz, 1H), 7.43 (t, J=7.6 Hz, 2H), 4.96 (d, J=5.0 Hz, 1H), 1.22 (s, 6H), 0.96–0.85 (m, 6H), 0.72–0.64 (m, 3H). ¹⁹ F NMR (377 MHz, CDCl 3 ) δ-110.61.

Compound 66-5': ¹⁹ F NMR (377 MHz, CDCl3) δ -88.95, -89.58, -110.84, -111.47.

Step 6: Dissolve compound (1R, 3aR, 7S, 9aS, 9bR, 11aR)-4-fluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethyl-2, 3, 3a, 5, 5a, 6, 7, 8, 9, 9a, 9b, 10, 11, 11a-tetradecahydro-1H-cyclopenta[1, 2-a]phenanthrene-7-ol 66-5 (100 mg, 0.2 mmol, 1 eq) in tetrahydrofuran (4 mL) and methanol (2 ml), add lithium hydroxide monohydrate (82 mg, 2 mmol, 10 eq), stir at 40 degrees for 3 hours, detect by TLC, and add after the reaction is completed. The reaction mixture was quenched with water, extracted with ethyl acetate (50 mL), and the organic phase was washed twice with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and dried to give a crude product, which was then separated and purified on silica gel (petroleum ether: ethyl acetate = 1:1) to give (1R, 3aR, 7S, 9aS, 9bR, 11aR)-4-fluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol 66 (70 mg, 0.16 mmol, yield: 85%).

Compound 66: ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.60 (t, J = 4.4 Hz, 1H), 2.08-1.90 (m, 3H), 1.83 (dddd, J = 19.8, 15.3, 9.9, 5.8 Hz, 6H), 1.63 (s, 1H), 1.56-1.38 (m, 10H), 1.35-1.25 (m, 3H), 1.22 (s, 6H), 1.18-1.03 (m, 4H), 0.94 (d, J = 6.5 Hz, 3H), 0.82 (s, 3H), 0.64 (s, 3H).

Embodiment 79

Preparation of compound 79 (1R, 3aR, 5aR, 7S, 9aR, 9bR, 11aR)-4-fluoro-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethyl-2,3,3a, 5,5a, 6,7,8,9,9a,9b, 10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6-one.

Step 1: Dissolve (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-8 (1 g, 2.17 mmol) in anhydrous tetrahydrofuran (20 mL) in a 100 mL round-bottom flask at room temperature. The reaction mixture was cooled to 0°C under nitrogen protection, and methylmagnesium bromide (7.23 mL, 21.70mmol), then stirred at room temperature for 2 hours, monitored by TLC (petroleum ether: ethyl acetate = 2: 1), after the reaction, the system was cooled to 0 ° C, quenched with saturated aqueous ammonium chloride solution (60mL), extracted with ethyl acetate (50mL×3), the organic phase was washed with saturated brine (80mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain cholest-6(5)-ene-3β,4β,25-triol 79-1 (800mg, purity: 90%, yield: 79.38%) as a white solid. ¹ H NMR (400MHz, CDCl ₃ )δ5.68(d,J＝3.3Hz,1H),4.14(s,1H),3.56(d,J＝11.1Hz,1H),2.22–1.72(m,7H),1.72–1.3 1(m,13H),1.21(s,6H),1.18(s,3H),1.15–1.00(m,5H),0.93(d,J=6.5Hz,3H),0.68(s,3H).

Step 2: Cholest-6(5)-ene-3β,4β,25-triol 79-1 (400 mg, 0.96 mmol) was dissolved in tetrahydrofuran (15 mL) in a 100 mL round-bottom flask at room temperature. Triethylamine (2.9 g, 28.8 mmol), 4-dimethylaminopyridine (117 mg, 0.96 mmol) and acetic anhydride (980 mg, 9.6 mmol) were added at room temperature. The mixture was stirred at 80 °C for 24 hours and monitored by TLC (petroleum ether: ethyl acetate = 10:1). After the reaction was completed, the mixture was quenched with water (50 mL), extracted with ethyl acetate (30 mL × 3), and the organic phase was saturated with water. The reaction mixture was washed with brine (50 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1) to give white solid acetate-(6R)-6-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-diacetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylhept-2-yl ester 79-2 (415 mg, purity: 90%, yield: 71.76%). ¹ H NMR(400MHz, CDCl3)δ＝5.84–5.79(m,1H),5.50(d,J＝2.5,1H),4.74(dt,J＝7.8,4.1,1H),2.07(s,3H),2.05–2.02(m,1H),2.01(s,3H),1.97(s, 3H),1.93–1.54(m,10H),1.54–1.44(m,3H),1.42(s,6H),1.39–1.15(m ,7H),1.13(s,3H),1.10–0.96(m,4H),0.92(d,J＝6.5,3H),0.67(s,3H).

Step 3: In a 100 mL round-bottom flask at room temperature, (6R)-6-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-diacetoxy-9a,11a-dimethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylhept-2-yl acetate 79-2 (400 mg, 0.73 mmol) was dissolved in acetone (15 mL), and N-hydroxyphthalimide (24 mg, 0.15 mmol), cobalt acetate (13 mg, 0.073 mmol) and tert-butyl hydroperoxide (265 mg, 2.94 mmol) were added at room temperature, followed by stirring at room temperature for 36 hours. TLC (stone) showed a clear color. After the reaction was completed, the reaction was quenched with water (50 mL), extracted with ethyl acetate (40 mL×3), the organic phase was washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1) to obtain white solid acetate-(6R)-6-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-diacetoxy-9a,11a-dimethyl-4-oxyylidene-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylhept-2-yl ester 79-3 (300 mg, purity: 90%, yield: 65.81%). ¹ H NMR (400MHz, CDCl3) δ5.92(s,1H),5.61(d,J＝2.3Hz,1H),4.80(dt,J＝7.8,4.2Hz,1H),2.34(dt,J＝22.4,9.8Hz,2H),2.10(s,3H),2.03(s, 3H),1.97(s,3H),1.92–1.52(m,10H),1.42(s,6H),1.39–1.34(m,2H),1.32(s,3H),1.29–1.00(m,8H),0.93(d,J＝6.5Hz,3H),0.68(s,3H). ¹³ C NMR (101MHz, CDCl ₃ )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.

Step 4: In a 50 mL round-bottom flask at room temperature, add acetic acid-(6R)-6-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-diacetoxy- 9a,11a-Dimethyl-4-oxydeoxy-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylhept-2-yl ester 79-3 (300 mg, 0.54 mmol) was dissolved in ethyl acetate (10 mL) and tetrahydrofuran (10 mL), 10% Pd/C (150 mg) was added at room temperature, and the mixture was stirred in a hydrogen atmosphere at room temperature for 1 hour. The reaction was monitored by TLC (ethyl acetate/petroleum ether = 15:1). After the reaction, 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 = 15:1) to obtain white solid acetate-(6R)-6-[(1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-6,7-diacetoxy-9a, 11a-dimethyl-4-oxyylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylhept-2-yl ester 79-4 (200 mg, purity: 90%, yield: 59.60%). ¹ H NMR(400MHz, CDCl3)δ5.17(s,1H),4.83–4.74(m,1H),2.58–2.46(m,1H),2. 37–2.28(m,1H),2.25–2.17(m,1H),2.11(s,3H),2.07–2.02(m,1H),1.97(d, J＝4.3Hz,6H),1.94–1.81(m,3H),1.80–1.46(m,9H),1.40–1.31(m,5H),1.28 (s,3H),1.26(s,6H),1.16–1.01(m,5H),0.91(d,J＝6.5Hz,3H),0.64(s,3H). ^13C NMR (101MHz, CDCl ₃ )δ=210.87,170.51,170.19,82.52,72.07,71.97,55.16,54.96,49.99,48.91,48.58,42.77,42.53,41.15,38.50,36.17, 35.78,35.52,31.93,29.70,28.38,26.05,25.06,22.69,22.51,21.13,20.97,20.91,20.32,18.70,14.12,13.65,12.04.

Step 5: In a 50 mL round-bottom flask at room temperature, (6R)-6-[(1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-6,7-diacetoxy-9a, 11a-dimethyl-4-oxyylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylheptyl-2-yl acetate 79-4 (200 mg, 0.36 mmol) was dissolved in diethylaminosulfur trifluoride (3 mL), stirred at 40 °C for 6 hours, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 10: 1). After the reaction was completed, the reaction solution was added to ice water (30 mL) to quench, extracted with ethyl acetate (20 mL × 3), the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain a white solid ethyl acetate. Acetic acid-(6R)-6-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-diacetoxy-4,4-difluoro-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylheptyl ester 79-5A and acetate-(6R)-6-[(1R,3aR,5aR,6R,7S,9aR,9bS,11aR)-6,7-diacetoxy-4,4-difluoro-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylheptyl ester bR,11aR)-6,7-diacetoxy-4-fluoro-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylhept-2-yl ester mixture 79-5B mixture (120 mg, purity: 90%. Yield calculated based on 79-5A: 51.55%).

Compound 79-5A: ¹ H NMR(400MHz, CDCl3)δ5.20(s,1H),4.79(dd,J＝8.3,4.0Hz,1H),2.10(s,3H),1.97(d,J＝3.6Hz,6H),1 .92–1.59(m,21H),1.42(s,6H),1.19–1.09(m,6H),1.05(s,3H),0.91(d,J＝6.5Hz,3H),0.66(s,3H ). ^13C NMR (101MHz, CDCl3) δ170.25,82.53,72.01,71.84,55.25,43.05,41.15,39.11,36.14 ,35.61,29.70,26.05,22.67,22.50,20.98,20.92,20.55,20.36,18.66,13.25,11.83.

Step 6: Acetic acid-(6R)-6-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-4,4-difluoro-6,7-dihydroxy-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylhept-2-yl ester 79-5A and acetate-(6R)-6-[(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-6,7-diacetoxy-4-fluoro-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9 b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylheptyl-2-yl ester mixture 79-5B mixture (385 mg, 0.77 mmol) was dissolved in tetrahydrofuran (5 mL) and methanol (5 mL), lithium hydroxide monohydrate (324 mg, 7.72 mmol) and water (3 mL) were added at room temperature, and then stirred at 45°C for 16 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction was completed, it was diluted with water (50 mL), extracted with ethyl acetate (40 mL×3), and the organic phase was washed with saturated brine (80 mL), and anhydrous sulfuric acid was added. The mixture was dried over sodium sulfate, the organic phase was concentrated, and the mixture was purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1) to obtain white solids (1R,3aS,3bS,7S,9aR,9bS,11aR)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol 79-6A and (1R,3aR,5aR,6R,7S, A mixture of (9aR,9bR,11aR)-4-fluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol 79-6B (340 mg, purity: 90.18%, yield calculated based on 79-6A: 86.97%).

Compound 79-6A: ¹ H NMR (400 MHz, CDCl3) δ = 3.74 (s, 1H), 3.63–3.57 (m, 1H), 2.32–1.72 (m, 15H), 1.57–1.25 (m, 14H), 1.21 (s, 6H), 1.06 (s, 3H), 0.93 (d, J = 6.5, 3H), 0.67 (s, 3H).

Step 7: In a 50 mL round-bottom flask at room temperature, (1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol 79-6A and (1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-4-fluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol 79 -6B mixture (1.41 g, 3.09 mmol) was dissolved in dichloromethane (30 mL), triethylamine (0.94 g, 9.26 mmol), 4-dimethylaminopyridine (0.3 g, 2.47 mmol) and benzoyl chloride (0.65 g, 4.63 mmol) were added at room temperature, and then stirred at room temperature for 20 minutes. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 4:1). After the reaction was completed, it was quenched with water (100 mL), extracted with ethyl acetate (60 mL×3), the organic phase was washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) was performed to obtain a white solid (1.5 g) which was subjected to chiral separation (instrument: Waters Acquity UPCC; column: REGIS CHIRAL (S, S)-whey O1 4.6*150 mm, 3.5 um; mobile phase: A/B: CO2/MeOH (0.1% DEA) = 50/50; flow rate: 1.5 ml/min; column temperature: 37 degrees) to obtain a white solid (1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-7-(benzyloxy)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-ol 79-7A (1.04 g, purity: 95%, yield: 58.52%, retention time t _R =2.597 min) and (1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-7-(benzyloxy)-4-fluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6-ol 79-7B (0.24 g, purity: 95%, yield: 14.02%, retention time t _R =2.963 min).

Compound 79-7A: ¹ H NMR (400 MHz, CDCl3) δ = 8.11–7.97 (m, 2H), 7.58 (t, J = 7.4, 1H), 7.46 (t, J = 7.7, 2H), 4.99 (dt, J = 7.3, 3.9, 1H), 3.98 (s, 1H), 2.46–1.23 (m, 25H), 1.22 (s, 6H), 1.14 (s, 3H), 1.12–0.97 (m, 3H), 0.94 (d, J = 6.5, 3H), 0.68 (s, 3H). ¹⁹ F NMR (376 MHz, CDCl3) δ = -89.02 (d, J = 236.3, 1F), -110.79 (d, J = 236.2, 1F).

Compound 79-7B: ¹ H NMR (400 MHz, CDCl3) δ = 8.05 (t, J = 6.1, 2H), 7.58 (t, J = 7.4, 1H), 7.46 (t, J = 7.7, 2H), 5.03-4.95 (m, 1H), 4.10 (d, J = 15.7, 1H), 2.73 (dd, J = 16.8, 8.5, 1H), 2.40-1.29 (m, 26H), 1.22 (s, 6H), 1.13 (s, 3H), 0.95 (d, J = 6.4, 3H), 0.65 (s, 3H). ¹⁹ F NMR (376 MHz, CDCl3) δ = -109.82 (s, 1F).

Step 8: In a 50 mL round-bottom flask at room temperature, (1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-7-(benzyloxy)-4-fluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6-ol 79-7B (100 mg, 0.19 mmol) was dissolved in dichloromethane (10 mL). Dess-Martin periodinane (242 mg, 0.57 mmol) was added at room temperature. mmol), stirred at room temperature for 2 hours. The reaction was monitored by TLC (ethyl acetate/petroleum ether=4:1). After the reaction, saturated aqueous sodium sulfite solution (5 mL) and saturated aqueous sodium bicarbonate solution (5 mL) were added to quench the reaction, and the mixture was extracted with dichloromethane (10 mL×3). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated. The organic phase was purified by silica gel column chromatography (petroleum ether:ethyl acetate=4:1) to obtain a white solid (1R,3aR,5aR,7S,9aR,9bR,11aR)-7-(benzyloxy)-4-fluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6-one 79-8 (81 mg, purity: 80%, yield: 65.05%). ¹ H NMR (400MHz, CDCl3) δ = 8.10 (d, J = 7.4, 2H), 7.58 (t, J = 7.4, 1H), 7.45 (t, J = 7.7, 2H), 5.48–5.41 (m, 1H ),2.64–2.33(m,3H),2.26–1.30(m,23H),1.22(s,6H),0.95(d,J=6.5,3H),0.82(s,3H),0.64(s,3H). ^13C NMR (101MHz, CDCl3) δ = 203.65, 165.72, 133.24, 129.90, 128.38, 71.12, 54.36, 53.16, 52.49, 48.80, 44.37, 43.60, 41.83, 41. 17,39.43,36.44,36.19,35.61,29.41,29.37,29.22,28.49,28.34,25.12,25.03,23.37,22.28,20.84,18.97,14.97,12.61. ¹⁹ F NMR (376MHz, CDCl3) δ = -110.02 (s, 1F).

Step 9: In a 50 mL round-bottom flask at room temperature, (1R,3aR,5aR,7S,9aR,9bR,11aR)-7-(benzyloxy)-4-fluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6-one 79-8 (80 mg, 0.15 mmol) was dissolved in tetrahydrofuran (2 mL) and methanol (2 mL), potassium carbonate (105 mg, 0.76 mmol) and water (1 mL) were added at room temperature, followed by stirring at room temperature for 4 hours. The mixture was detected by TLC (petroleum ether:ethyl acetate=3:1). The reaction was tested. After the reaction was completed, the mixture was diluted with water (20 mL), extracted with ethyl acetate (10 mL×3), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1) to give a white solid (1R, 3aR, 5aR, 7S, 9aR, 9bR, 11aR)-4-fluoro-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6-one 79 (15.55 mg, purity: 100%, yield: 23.47%).

Compound 79: ¹ H NMR (400 MHz, CDCl3) δ = 4.13 (dd, J = 17.4, 7.6, 1H), 2.60-2.35 (m, 3H), 2.19-1.27 (m, 24H), 1.22 (s, 6H), 0.94 (d, J = 6.3, 3H), 0.74 (s, 3H), 0.64 (s, 3H). ¹³ C NMR(101MHz, CDCl3)δ=210.89,74.90,71.11,54.35,52.51,52.30,52.21,48.94,48.87,44.37,43.65,41.71,39.42,36.43,36.17 ,35.03,32.02,30.13,29.71,29.43,29.21,28.48,28.46,25.10,25.02,23.40,23.10,22.24,22.23,20.82,18.96,15.04,12.60. ^19F NMR (376MHz, CDCl3) δ = -110.10 (s, 1F). LC-MS: [M+Na] ⁺ = 457.1

Embodiment 85 & 86

Compound 85 or 86 7β-fluoro-5α-cholest-3β,25-diol

Preparation of compound 86 or 85 7α-fluoro-5α-cholest-3β,25-diol.

In the first step, (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-7 (1 g, 2.25 mmol, 1 eq) was dissolved in tetrahydrofuran (10 mL), and cobalt acetate (80 mg, 0.45 mmol, 0.2 eq), N-hydroxyphthalimide (0.15 g, 0.90 mmol, 0.4 eq) and tert-butyl peroxide (1.01 g, 11.25 mmol, 5 eq) were added to the solution and stirred at room temperature for 20 hours. After the reaction was completed, 30 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL*3). The obtained organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product. The crude product was purified by silica gel column (petroleum ether/ethyl acetate = 10/1) to obtain (5R)-5-[(1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-7-acetoxy-9a, 11a-dimethyl-4-oxyylidene-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 72-1 (0.6 g, 1.31 mmol, yield: 58%). ¹ H NMR (400 MHz, CDCl ₃ )δ5.70(d,J＝1.5Hz,1H),4.80–4.64(m,1H),3.67(s,3H),2.48(dddd,J＝15.6,12.1,9.1,2.4 Hz,3H),2.25(ddd,J＝22.3,12.9,5.3Hz,3H),2.05(s,3H),2.01–1.95(m,2H),1.93–1.84(m, 1H),1.79–1.65(m,2H),1.65–1.46(m,5H),1.40(ddd,J＝11.6,8.9,4.3Hz,2H),1.28(ddd,J＝ 10.9, 10.5, 5.9Hz, 3H), 1.21 (s, 3H), 1.18–1.05 (m, 3H), 0.94 (d, J = 6.5Hz, 3H), 0.68 (s, 3H).

In the second step, compound (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-4-oxyylidene-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 72-1 (0.6 g, 1.31 mmol, 1 eq) was dissolved in ethyl acetate (15 mL), and then palladium carbon (10%) (120 mg) was added thereto and stirred at room temperature for 12 hours. Filter and spin dry the organic solvent to obtain the product (5R)-5-[(1R,3aS,3bR,7S,9aS,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-4-oxyylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 72-2 (450 mg, 0.977 mmol, yield: 75%) as an oily compound. ¹ H NMR (400 MHz, cdcl ₃ )δ5.36–5.24(m,1H),4.30(s,3H),3.00–2.81(m,5H),2.70–2.58(m,5H),2.55–2.4 7(m,2H),2.42(dt,J＝13.4,3.5Hz,1H),2.35–2.26(m,2H),2.22–2.10(m,6H),2.09– 1.99(m,3H),1.86(d,J＝12.4Hz,1H),1.76(s,1H),1.73(s,1H),1.73(s,3H),1.70(s ,1H),1.69(d,J＝3.3Hz,1H),1.67–1.60(m,1H),1.56(d,J＝6.5Hz,3H),1.28(s,3H).

The third step: (5R)-5-[(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-4-oxyylidenehexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 72-2 (300 mg, 0.65 mmol, 1 eq) was dissolved in methanol (10 mL), sodium borohydride (30 mg, 0.78 mmol, 1.2 eq) was added to the solution, and stirred at room temperature for 1 hour. After the reaction was completed, 30 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL*3). The obtained organic phase was dried over anhydrous sodium sulfate and then spin-dried to give a crude product of (5R)-5-[(1R, 3aS, 3bR, 5aS, 7S, 9aS, 9bS, 11aR)-7-acetoxy-4-hydroxy-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 85-1 (240 mg, 0.52 mmol, yield: 80%), which was directly used for the next step.

Step 4: Compound (5R)-5-[(1R, 3aS, 3bR, 5aS, 7S, 9aS, 9bS, 11aR)-7-acetoxy-4-hydroxy-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 85-1 (240 mg, 0.52 mmol, 1 eq) was dissolved in ultra-dry pyridine (5 mL), and then 4-methylbenzenesulfonyl chloride (824 mg, 4.32 mmol, 10 eq) was added thereto. The mixture was stirred at room temperature for 5 days. After the reaction was completed, 30 ml of 3M aqueous hydrochloric acid solution was added, and the mixture was extracted with ethyl acetate (30 mL*3). The organic solvent was then dried to obtain a crude product. The crude product was separated and purified by silica gel separation (petroleum ether: ethyl acetate = 10:1) to obtain the product, which was separated by SFC (instrument: Waters Acquity UPCC; chromatographic column: Daicel CHIRALPAK IB 4.6*250mm, 5um; mobile phase: A/B:CO2/MeOH (0.1% DEA) = 60/40; flow rate: 1.5 ml/min; column temperature: 37 degrees) to obtain 85-2 (60 mg, 0.1 mmol, yield: 22.5%, retention time t _R = 1.769 min) and 86-1 (70 mg, 0.13 mmol, yield: 30%, retention time t _R = 2.792 min) as oily compounds.

Compound 85-2: ¹ H NMR (400 MHz, CDCl ₃ ) δ7.76 (d, J=8.1 Hz, 1H), 7.31 (d, J=8.1 Hz, 1H), 4.67–4.57 (m, 0H), 4.46 (td, J=10.4, 5.1 Hz, 0H), 3.67 (s, 3H), 2.44 (s, 3H), 2.26 (dd, J=15.3, 8.2 Hz, 2H), 2.00 (s, 3H), 1.96 (d, J=13.0 Hz, 1H), 1.8 1(d,J＝15.1Hz,3H),1.73–1.63(m,4H),1.53(s,3H),1.47(d,J＝9.6Hz,2H),1.41–1.32(m,4H),1. 28–1.17(m,5H),1.14–0.99(m,4H),0.91(d,J＝6.4Hz,3H),0.81(s,3H),0.76(s,1H),0.62(s,3H).

Compound 86-1: ¹ H NMR (400 MHz, CDCl ₃ ) δ7.78 (d, J=8.1 Hz, 2H), 7.35 (d, J=8.0 Hz, 2H), 4.76 (s, 1H), 4.62–4.46 (m, 1H), 3.68 (s, 3H), 2.45 (s, 3H), 2.26 (dt, J=15.7, 7.7 Hz, 2H), 2.00 (s, 3H), 1.90 (d, J=13.0 Hz, 1H), 1.79 (d, J=1 3.2Hz,1H),1.69(d,J＝12.4Hz,2H),1.46(dd,J＝17.7,12.2Hz,6H),1.38(d,J＝13.0Hz,3H),1 .29–1.17(m,6H),1.08(dt,J＝22.8,12.9Hz,6H),0.90(d,J＝6.3Hz,3H),0.77(s,3H),0.60(s, 3H).

Step 5: 85-2 (60 mg, 0.1 mmol, 1.0 eq) and TBAF (38 mg, 0.146 mmol, 1.5 eq) were dissolved in tetrahydrofuran (2 mL), stirred at 65 °C for 12 hours, and detected by TLC. After the reaction was completed, water was added to quench the reaction, and then extracted with ethyl acetate (50 mL). The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried to obtain a crude product, which was then separated and purified on silica gel (petroleum ether: ethyl acetate = 1:1) to obtain 85-3 (20 mg, 0.145 mmol, yield: 44%), which was directly used for the next step.

Step 6: 85-3 (20 mg, 0.043 mmol, 1 eq) was dissolved in tetrahydrofuran (2 mL), and methylmagnesium bromide (2.4 M) (0.6 mL, 10 eq) was slowly added dropwise after replacing nitrogen. The mixture was stirred at room temperature for 3 hours and tested by TLC. After the reaction was completed, water was added to quench the mixture, and then the mixture was extracted with ethyl acetate (50 mL). The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried to obtain a crude product, which was then separated and purified on silica gel (petroleum ether: ethyl acetate = 4:1) to obtain 85 (6 mg, 0.014 mmol, yield: 33%).

Compound 85: ¹ H NMR (400 MHz, CDCl ₃ )δ4.63(d,J＝49.6Hz,1H),3.63(s,1H),1.95(d,J＝12.6Hz,1H),1.88–1.80( m,2H),1.76–1.68(m,3H),1.63(d,J＝13.2Hz,2H),1.52(dd,J＝11.0,8.1Hz,6 H),1.39(dd,J＝12.3,9.6Hz,7H),1.29–1.25(m,4H),1.21(s,6H),1.10(ddd ,J＝14.2,10.1,5.1Hz,4H),0.92(d,J＝6.5Hz,3H),0.80(s,3H),0.65(s,3H). ¹⁹ F NMR (376MHz, CDCl ₃ )δ-196.28.

Similar operation is performed to replace raw material 85-2 with 86-1 to obtain 86.

Compound 86: ¹ H NMR (400MHz, CDCl ₃ )δ4.28–4.09(m,1H),3.65–3.54(m,1H),2.00(dd,J＝9.4,3.4Hz,1H),1. 86–1.70(m,5H),1.59(d,J＝11.5Hz,2H),1.52(d,J＝12.9Hz,2H),1.45(d, J＝10.9Hz,4H),1.35(dd,J＝12.4,4.5Hz,5H),1.30–1.23(m,3H),1.21(s, 6H),1.17–1.03(m,5H),0.92(d,J＝6.5Hz,3H),0.86(s,3H),0.68(s,3H). ^19F NMR (376MHz, CDCl ₃ ) δ-167.50.

Embodiment 94

Preparation of Compound 94 (1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4,4-difluoro-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol

Step 1: At room temperature, (R)-5-((3S, 4R, 8S, 9S, 10R, 13R, 14S, 17R)-3-acetoxy-4-hydroxy-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-17-yl)-2,2-difluorohexanoic acid ethyl ester 34-1 (450 mg, 0.88 mmol) was dissolved in anhydrous tetrahydrofuran (12 mL). The temperature was lowered to 0°C under nitrogen protection, and methylmagnesium bromide (3 M in ) was added dropwise. 2-Me-THF, 2.9 mL, 8.8 mmol), then stirred at room temperature for 2 hours, monitored by TLC (petroleum ether: ethyl acetate = 2: 1), after the reaction was completed, the system was cooled to 0 ° C, quenched with saturated aqueous ammonium chloride solution (30 mL), extracted with ethyl acetate (30 mL × 2), the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1) to obtain a white solid (3S, 4R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-5,5-difluoro-6-hydroxy-6-methylheptane-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3,4-diol 94-1 (195 mg, purity: 95%, yield: 49%). ¹ H NMR (400MHz, CDCl ₃ )δ5.68(d,J＝3.4Hz,1H),4.14(d,J＝3.4Hz,1H),3.57(dd,J＝7.7,4.0Hz,1H ),2.14–1.94(m,4H),1.94–1.79(m,4H),1.74(d,J＝18.2Hz,1H),1.68–1.5 3(m,6H),1.52–1.40(m,4H),1.31(s,6H),1.18(s,3H),1.16–1.07(m,3H), 1.05–0.98(m,1H),0.94(d,J＝6.6Hz,3H),0.90–0.82(m,1H),0.69(s,3H).

In the second step, 3S, 4R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-5,5-difluoro-6-hydroxy-6-methylheptane-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopentadien[a]phenanthrene-3,4-diol 94-1 (193 mg, 0.4 2mmol, 1.0eq) was dissolved in tetrahydrofuran (10mL), triethylamine (6.37g, 63.0mmol, 150eq), 4-dimethylaminopyridine (154mg, 1.26mmol, 3.0eq) and acetic anhydride (2.14g, 21.0mmol, 50eq) were added at room temperature, and then stirred at 80°C for 24 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1:1). After the reaction was completed, it was quenched with ice water (50mL), ethyl acetate (30mL×2) was added, and the mixture was stirred for 24 hours at 80°C. After extraction, the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1) to give a white solid (3S, 4R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-6-acetoxy-5,5-difluoro-6-methylheptane-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopentadienyl[a]phenanthrene-3,4-diyl diacetate 94-2 (182 mg, purity: 90%, yield: 74.59%).

¹ H NMR (400MHz, CDCl ₃ )δ5.84–5.79(m,1H),5.50(d,J＝2.6Hz,1H),4.74(dt,J＝12.4,4.1Hz,1H),2.15–2 .08(m,1H),2.07(s,3H),2.03(s,3H),2.01(s,3H),1.97–1.65(m,6H),1.60(s,6H) ,1.48(d,J＝9.6Hz,3H),1.27(d,J＝12.1Hz,7H),1.23–1.16(m,2H),1.13(s,3H),1. 13–0.97(m,3H),0.95(d,J＝6.5Hz,3H),0.88(dd,J＝13.4,7.8Hz,1H),0.69(s,3H).

Step 3: (3S, 4R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-6-acetoxy-5,5-difluoro-6-methylheptane-2-yl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopentadienyl[a]phenanthrene-3,4-diyl diacetate 94-2 (182 mg, 0.31 mmol, 1.0 eq) was dissolved in acetone (8 mL) at room temperature, and N-hydroxyphthalimide (10 mg, 0.062 mmol, 0.2 eq), cobalt acetate (5.5 mg, 0.031 mmol, 0.1 eq) and tert-butyl hydroperoxide (5 M in ) were added at room temperature. hexane, 0.25 mL, 1.24 mmol, 4.0 eq), and then stirred at room temperature for 36 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 10: 1). After the reaction was completed, it was quenched with water (30 mL), extracted with ethyl acetate (20 mL × 2), and the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15: 1) to obtain a white solid (3S, 4 R,8S,9S,10R,13R,14S,17R)-17-((R)-6-acetoxy-5,5-difluoro-6-methylheptan-2-yl)-10,13-dimethyl-7-oxyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3,4-diyl diacetate 94-3 (120 mg, purity: 90%, yield: 65.22%). ¹ H NMR (400MHz, CDCl ₃ )δ5.93(s,1H),5.61(d,J＝2.7Hz,1H),4.80(dt,J＝12.2,4.0Hz,1H),2.43–2.26(m,2H),2.09(d,J＝5.7Hz,4H),2.04(d,J＝5.4Hz ,7H),2.00–1.87(m,3H),1.85–1.66(m,3H),1.59(s,6H),1.48(s,2H),1.39–1.04(m,12H),0.96(d,J=6.5Hz,3H),0.70(s,3H). ¹⁹ F NMR (376MHz, CDCl ₃ )δ-113.92–-117.67(m,2F).

Step 4: (3S, 4R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-6-acetoxy-5,5-difluoro-6-methylheptane-2-yl)-10,13-dimethyl-7-oxy-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3,4-diyl diacetate 94-3 (120 mg, 0.20 mmol) was dissolved in ethyl acetate (3 mL) and tetrahydrofuran (3 mL) at room temperature, 10% Pd/C (60 mg) was added at room temperature, and the mixture was stirred in a hydrogen atmosphere at room temperature for 6 hours. The reaction was monitored by TLC (ethyl acetate/petroleum ether=1:4). After the reaction, 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 = 15:1) to obtain a white solid (3S, 4R, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-6-acetoxy-5,5-difluoro-6-methylheptane-2-yl)-10,13-dimethyl-7-oxohexadecahydro-1H-cyclopentadienyl[a]phenanthrene-3,4-diyl diacetate 94-4 (53 mg, purity: 90%, yield: 44.54%). ¹ H NMR (400 MHz, CDCl ₃ )δ5.17(s,1H),4.78(dd,J＝12.1,3.6Hz,1H),2.52(t,J＝13.5Hz,1H),2.34(t,J＝11. 4Hz,1H),2.27–2.18(m,1H),2.12(s,3H),2.10–2.05(m,1H),2.03(s,3H),1.98(s,3 H),1.96–1.82(m,4H),1.71(t,J＝12.8Hz,4H),1.59(s,6H),1.44(ddd,J＝18.3,17.6 ,6.7Hz,4H),1.35–1.24(m,6H),1.19–1.04(m,4H),1.01–0.91(m,4H),0.66(s,3H).

Step 5: (3S, 4R, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-6-acetoxy-5,5-difluoro-6-methylheptane-2-yl)-10,13-dimethyl-7-oxohexadecahydro-1H-cyclopentadienyl[a]phenanthrene-3,4-diyl diacetate 94-4 (53 mg, 0.089 mmol) was dissolved in diethylaminosulfur trifluoride (2 mL) at room temperature and stirred at 70 ° C for 2 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 4:1). After the reaction was completed, the reaction solution was diluted with ethyl acetate and slowly added dropwise to ice water (20 mL ) was quenched, extracted with ethyl acetate (15 mL×2), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1) to give a white solid (3S, 4R, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-6-acetoxy-5,5-difluoro-6-methylheptane-2-yl)-7,7-difluoro-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-3,4-diyl diacetate 94-5 (45 mg, purity: 90%, yield: 81.82%). ¹ H NMR (400MHz, CDCl ₃ )δ5.21(s,1H),4.83–4.76(m,1H),2.10(s,3H),2.03(s,3H),1.98(s,3H), 1.83(dd,J＝14.1,10.0Hz,5H),1.76–1.67(m,4H),1.59(s,6H),1.46(dd,J ＝18.1,9.1Hz,4H),1.30(dd,J＝15.2,7.1Hz,4H),1.14(d,J＝12.8Hz,3H),1 .06(s,3H),0.99(s,1H),0.95(d,J＝6.5Hz,3H),0.88(s,1H),0.68(s,3H). ^19F NMR (377MHz, CDCl ₃ ) δ -89.25 (d, J = 238.2Hz, 1F), -110.61 (d, J = 237.8Hz, 1F), -114.63–-116.99 (m, 2F).

Step 6: (3S, 4R, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-6-acetoxy-5,5-difluoro-6-methylheptane-2-yl)-7,7-difluoro-10,13-dimethylhexahydro-1H-cyclopentadienyl[a]phenanthrene-3,4-diyl diacetate 94-5 (45 mg, 0.073 mmol, 1.0 eq) was dissolved in tetrahydrofuran (1 mL) and methanol (1 mL) at room temperature, and lithium hydroxide monohydrate (15 mg, 0.365 mmol, 5.0 eq) and water (0.5 mL) were added at room temperature, followed by stirring at room temperature for 30 min, and TLC (petroleum ether: acetic acid) was performed. After the reaction was completed, the mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL×2), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1) to obtain a white solid (3S, 4R, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-5,5-difluoro-6-hydroxy-6-methylheptane-2-yl)-7,7-difluoro-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-3,4-diol 94-6 (28 mg, purity: 90%, yield: 77.78%). ¹ H NMR (400MHz, CDCl ₃ )δ3.74(s,1H),3.66–3.56(m,1H),2.32–2.11(m,1H),1.98(d,J＝12.5Hz ,2H),1.91–1.77(m,6H),1.76–1.64(m,6H),1.44(dd,J＝17.3,4.5Hz,5H) ,1.30(s,6H),1.12(dd,J＝17.0,12.1Hz,3H),1.06(d,J＝5.4Hz,3H),1.00 (d,J＝4.0Hz,1H),0.94(d,J＝6.5Hz,3H),0.90–0.83(m,1H),0.68(s,3H).

Step 7: (3S, 4R, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-5,5-difluoro-6-hydroxy-6-methylheptane-2-yl)-7,7-difluoro-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-3,4-diol 94-6 (28 mg, 0.057 mmol, 1.0 eq) was dissolved in dichloromethane (2 mL) and tetrahydrofuran (1 mL) at room temperature, and triethylamine (35 mg, 0.34 mmol, 6.0 eq) was added at room temperature. , 4-dimethylaminopyridine (7 mg, 0.057 mmol, 1.0 eq) and benzoyl chloride (24 mg, 0.17 mmol, 3.0 eq), then stirred at room temperature for 8 hours, TLC (petroleum ether: ethyl acetate = 1: 1) monitored the reaction, after the reaction was completed, quenched with water (10 mL), extracted with ethyl acetate (10 mL × 2), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) and SFC separation (instrument: Waters Acquity UPCC; chromatographic column: Daicel CHIRALPAK IB 4.6*250 mm, 5 um; mobile phase: A/B: CO2/MeOH (0.1% DEA) = 60/40; flow rate: 1.5 ml/min; column temperature: 37 50% pyrrolidone) was added to give a white solid (3S,4R,5R,8S,9S,10R,13R,14S,17R)-17-((R)-5,5-difluoro-6-hydroxy-6-methylheptane-2-yl)-7,7-difluoro-4-hydroxy-10,13-dimethylhexahydro-1H-cyclopentyl[a]phenanthrene-3-ylbenzoate 94-7 (15 mg, purity: 95%, yield: 44.12%, retention time t _R =1.225 min). ¹ H NMR (400MHz, CDCl ₃ )δ8.04(d,J＝7.3Hz,2H),7.58(t,J＝7.4Hz,1H),7.46(t,J＝7.6Hz,2H),4.99 (d,J＝12.0Hz,1H),3.99(s,1H),2.37–2.16(m,1H),2.15–1.96(m,3H),1.83 (d,J＝9.5Hz,7H),1.76(s,3H),1.51–1.42(m,3H),1.32(s,3H),1.31(s,6H) ,1.14(s,2H),1.14(s,3H),1.03(s,2H),0.94(d,J=6.5Hz,3H),0.69(s,3H).

Step 8: (3S, 4R, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-5,5-difluoro-6-hydroxy-6-methylheptane-2-yl)-7,7-difluoro-4-hydroxy-10,13-dimethylhexahydro-1H-cyclopentyl[a]phenanthrene-3-ylbenzoate 94-7 (15 mg, 0.025 mmol, 1.0 eq) was dissolved in tetrahydrofuran (0.5 mL) and methanol (0.5 mL) at room temperature, and lithium hydroxide monohydrate (11 mg, 0.25 mmol, 10.0 eq) and water (0.2 mL) were added at room temperature, followed by stirring at room temperature for 30 min, and TLC (petroleum ether: ethyl acetate) was performed. After the reaction was completed, the mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL×2), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=4:1) to obtain a white solid (3S, 4R, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-5,5-difluoro-6-hydroxy-6-methylheptane-2-yl)-7,7-difluoro-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-3,4-diol 94 (6.23 mg, purity: 100%, yield: 50.89%).

Compound 94: ¹ H NMR (400 MHz, CDCl ₃ )δ3.76–3.72(m,1H),3.64–3.56(m,1H),2.27–2.12(m,1H),2.02–1.94(m, 2H),1.89–1.83(m,2H),1.82–1.78(m,2H),1.77–1.67(m,5H),1.67–1.57(m ,3H),1.48–1.39(m,3H),1.38–1.33(m,2H),1.32–1.27(m,7H),1.17–1.09( m,2H),1.06(s,3H),1.02–0.97(m,1H),0.94(d,J＝6.5Hz,3H),0.68(s,3H). ¹⁹ F NMR (377MHz, CDCl ₃ ) δ -88.97 (d, J = 235.8Hz, 1F), -110.95 (d, J = 235.7Hz, 1F), -114.05–-117.90 (m, 2F).

Embodiment 103

Compound 103 Preparation of 3β,25-dihydroxy-5α-cholestane-7-carbonitrile

In the first step, acetic acid-(1R,3aS,3bR,7S,9aS,9bS,11aR)-4-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester (200 mg, 0.432 mmol, 1 eq) was dissolved in ultra-dry pyridine (5 mL), and then 4-methylbenzenesulfonyl chloride (824 mg, 4.32 mmol, 10 eq) was added thereto. The mixture was stirred at room temperature for 5 days. After the reaction was completed, 30 ml of 3M hydrochloric acid solution was added. The mixture was extracted with ethyl acetate (30 mL * 3), and the organic solvent was then spin-dried to obtain a crude product. The crude product was separated and purified by silica gel (petroleum ether: ethyl acetate = 10: 1) to obtain the product 4-methylbenzenesulfonic acid-(1R, 3aS, 3bR, 5aR, 7S, 9aS, 9bS, 11aR)-7-acetoxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-4-yl ester (30 mg, 0.049 mmol, yield: 11%) as a white compound.

¹ H NMR (400MHz, CDCl ₃ )δ7.77(t,J＝7.9Hz,2H),7.33(dd,J＝17.9,8.2Hz,2H),4.76–4.44(m,2H),2.44(s,3H),2.00(s,3H ),1.77(ddd,J＝32.4,24.9,8.1Hz,8H),1.52– 1.32(m,13H),1.29–1.24(m,4H),1.22(d,J＝4.4Hz,6H),1.15–1.02(m,5 H), 0.90 (d, J = 6.4Hz, 3H), 0.79 (d, J = 15.5Hz, 3H), 0.62 (d, J = 7.6Hz, 3H).

In the second step, compound 4-methylbenzenesulfonic acid-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-7-acetoxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-4-yl ester 103-1 (60 mg, 0.1 mmol, 1.0 eq) and sodium cyanide (20 mg, 0.4 mmol, 4 eq) were dissolved in DMF (2 mL), stirred at 65 ° C for 12 hours, monitored by TLC (petroleum ether: ethyl acetate = 1:1), and water was added after the reaction was completed. The reaction mixture was quenched and then extracted with ethyl acetate (50 mL). The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then separated and purified on silica gel (petroleum ether: ethyl acetate = 1:1) to give acetic acid-(1R, 3aS, 3bR, 7S, 9aS, 9bS, 11aR)-4-cyano-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 103-2 (25 mg, 0.05 mmol, yield: 55%), and the crude product was directly put into the next step.

Step 3: Compound acetic acid-(1R,3aS,3bR,7S,9aS,9bS,11aR)-4-cyano-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 103-2 (25 mg, 0.05 mmol, 1 eq) was dissolved in methanol (2 mL), and potassium carbonate (29 mg, 0.212 mmol, 4 eq) was added. The mixture was stirred at room temperature for 3 hours and monitored by TLC (petroleum ether: ethyl acetate = 4:1). After the reaction was completed, water was added to quench the reaction, followed by extraction with ethyl acetate (50 mL). The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product, which was then separated and purified on silica gel (petroleum ether: ethyl acetate = 4:1) to obtain 3β,25-dihydroxycholest-7-carbonitrile 103 (16 mg, 0.037 mmol, yield: 70%).

Compound 103: ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.67 (s, 1H), 2.84 (s, 1H), 2.00–1.95 (m, 1H), 1.92–1.81 (m, 2H), 1.75–1.67 (m, 2H), 1.59 (s, 2H), 1.54 (s, 2H), 1.46–1.37 (m, 8H), 1.33–1.27 (m, 5H), 1.22 (s, 6H), 1.19 (s, 1H), 1.13–1.01 (m, 5H), 0.92 (d, J=6.5 Hz, 3H), 0.81 (d, J=4.2 Hz, 3H), 0.66 (s, 3H). ¹³ C NMR (101 MHz, CDCl ₃ )δ121.18,71.10,70.63,55.76,52.83,49.11,44.38,42.74,40.40,39.01,37.16,36.36,36.28,3 5.61,35.49,32.36,31.14,30.80,29.29,29.27,28.04,23.60,20.89,20.66,18.57,12.03,11.92.

Embodiment 106

Compound 106 Preparation of 24-[Hydroxy(2-methoxyphenyl)methyl]-7-methylidene-5α-cholane-3β,4β-diol

Step 1: Dissolve the raw material 1-bromo-2-methoxybenzene (7.64 g, 40.82 mmol, 3.5 eq) in anhydrous tetrahydrofuran (50 mL), cool the system to -78 ° C, add n-butyl lithium (14.0 mL, 34.99 mmol, 3.0 eq), and maintain the temperature and stir for 30 minutes. Compound (R)-5-(((3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-2,2,5a,7a-tetramethyl-3a,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetrahydro-4H-cyclopentyl[7,8]phenanthrene[1,2-d][1,3]dioxy-8-yl)hexanal III (5 g, 11.66 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL) and added to the above reaction solution. The system was stirred at -78°C for 30 min. TLC (petroleum ether: ethyl acetate = 5:1) was used to monitor the reaction progress, and the raw materials were completely consumed. 10 mL of water was added to the reaction system, extracted with ethyl acetate (100 mL × 3), the combined organic phases were washed with saturated sodium chloride (100 mL), the combined organic phases were dried, and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 90:10) to obtain a white solid (5R)-5-[(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR) -2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetrahydro-3aH-cyclopentadienyl[1',2':1,2]phenanthrene[7,8-d][1,3]dioxacyclo-8-yl]-1-(2-methoxyphenyl)hexan-1-ol 106-1 (4.9 g, purity 90%, yield 70.43%). ¹ H NMR (400 MHz, CDCl ₃ )δ7.32–7.28(m,1H),7.26–7.20(m,1H),7.02–6.82(m,2H),5.80(d,J＝2.7Hz,1H),4.85(dt,J＝9 .9,5.8Hz,1H),4.41(d,J＝5.8Hz,1H),4.10(dd,J＝13.6,6.4Hz,1H),3.84(d,J＝10.0Hz,3H),2.19 –1.98(m,3H),1.82–1.72(m,3H),1.63(dd,J＝13.2,8.6Hz,5H),1.53(s,4H),1.43(s,3H),1.35(s ,3H),1.28–1.23(m,1H),1.16(s,4H),1.13–1.01(m,25H),0.95–0.88(m,4H),0.72–0.64(m,3H).

Step 2: In a 200 mL round-bottom flask at room temperature, (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetrahydro-3aH-cyclopentadienyl[1',2':1,2]phenanthrene[7,8-d][1, 3] dioxacyclo-8-yl]-1-(2-methoxyphenyl)hexan-1-ol 106-1 (4.9 g, 9.13 mmol) was dissolved in dichloromethane (70 mL), tert-butyldimethylsilyl chloride (4.13 g, 27.39 mmol), 4-dimethylaminopyridine (450 mg, 3.65 mmol) and imidazole (2.49 g, 36.51 mmol) were added at room temperature, and then stirred at room temperature for 2 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 5:1). After the reaction was completed, it was diluted with water (100 mL), extracted with ethyl acetate (100 mL×3), the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain a white solid (9R)-9-[(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-tetrahydro-3aH-cyclopentadienyl[1',2':1,2]phenanthrene[7,8-d][1,3]dioxetan-8-yl]-5-( ² -methoxyphenyl)-2,3,3-tetramethyl-4-oxo-3-siladecane 106-2 (4.9 g, purity: 90%, yield: 74%). NMR (400MHz, CDCl3) δ7.60(d,J＝7.6Hz,1H),7.36–7.27(m,1H),7.11–7.01(m,1H),6.95(d,J＝8.1Hz,1H),5.94(d,J＝2. 7Hz,1H),5.21(dt,J＝7.5,3.9Hz,1H),4.55(d,J＝5.8Hz,1H),4.25(t,J＝5.8Hz,1H),3.95(s,3H),2.30–2.20(m,1H),2. 20–2.11(m,2H),1.96(ddd,J＝13.1,7.8,3.9Hz,1H),1.91–1.60(m,13H),1.58–1.45(m,7H),1.41(dd,J＝13.1,6.0Hz,3 H),1.36–1.26(m,5H),1.26–1.09(m,5H),1.08–1.01(m,12H),1.01–0.90(m,1H),0.87–0.78(m,3H),0.21–0.15(m,3H).

Step 3: In a 250 mL round-bottom flask at room temperature, (9R)-9-[(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-tetrahydro-3aH-cyclopentadienyl[1',2':1,2]phenanthrene[7,8-d][1,3]dioxacyclo[8- [4-(2-methoxyphenyl)-2,3,3-tetramethyl-4-oxo-3-siladecane 106-2 (4.9 g, 7.53 mmol) was dissolved in acetone (100 mL), and N-hydroxyphthalamide (370 mg, 2.26 mmol), cobalt acetate (200 mg, 1.13 mmol) and tert-butyl hydroperoxide (3.39 g, 37.63 mmol) were added at room temperature, followed by stirring at room temperature for 36 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 10: 1). After the reaction was completed, the mixture was quenched with water (50 mL), extracted with ethyl acetate (100 mL × 3), and the organic phase was washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain a white solid (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-8-[(9R) -5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxo-3-silyl]-2,2,5a,7a,7a-tetramethyl-4,5,5a,5b,6,7,8,9,10,10a,10b,11,12b-tetrahydro-3aH-cyclopenta[1':1,2]phenanthrene[7,8-d][1,3]dioxy-11,5-one 106-3 (2.6 g, purity: 90%, yield: 46.75%). ¹ H NMR (400 MHz, CDCl3)δ7.59(d,J＝7.5Hz,1H),7.37–7.27(m,1H),7.08(t,J＝7.4Hz,1H),6.95(d,J＝8.2Hz,1H ),6.06(s,1H),5.22(dd,J＝7.0,3.3Hz,1H),4.66(d,J＝6.4Hz,1H),4.47(q,J＝5.5Hz,1H),3.95 (s,3H),2.55–2.43(m,2H),2.24–2.14(m,1H),2.05–1.93(m,2H),1.81–1.67(m,11H),1.55–1 .39(m,15H),1.31–1.18(m,3H),1.06–0.99(m,13H),0.87–0.80(m,3H),0.16(q,J＝7.4Hz,3H).

Step 4: (3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladec-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-tetradecahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-11-one 106-3 (2.1 g, 3.158 mmol) was dissolved in methanol (30 mL), Pd/C (1.08 g, 5.052 mmol) was added, the system was replaced with hydrogen, and the reaction solution was stirred at room temperature for 2 hours. The reaction solution was filtered through diatomaceous earth, and the filtrate was dried by rotary evaporation and purified by silica gel column chromatography to obtain (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1', ² ':7,8]phenanthro[1,2-d][1,3]dioxolan-11-one 106-4 (1.1 g, 1.402 mmol, 44.39%) as a white solid. NMR (400MHz, CDCl3) δ7.45(d,J＝7.6Hz,1H),7.22–7.14(m,1H),6.94(t,J＝7.4Hz,1H),6.81(d,J＝8.0Hz,1H),5.06(s,1H),4.01(s,1H),4.00 –3.93(m,1H),3.81(s,3H),2.77(s,1H),2.42(s,1H),2.19(dd,J＝12.7,2.6Hz,2H),1.98(d,J＝12.9Hz,1H),1.93–1.82(m,2H),1.79–1.67(m ,2H),1.63(dd,J＝14.5,3.1Hz,2H),1.55(s,4H),1.48(d,J＝3.6Hz,2H),1.45–1.34(m,4H),1.30(s,6H),1.21(s,2H),1.06(dt,J＝11.1,8.1H z,4H),0.99(d,J＝5.6Hz,1H),0.96(d,J＝6.7Hz,1H),0.88(s,9H),0.64(d,J＝5.5Hz,3H),0.02(d,J＝1.8Hz,3H),-0.00(s,3H),-0.14(s,3H).

Step 5: Place methyltriphenylphosphonium bromide (696.19 mg, 1.949 mmol) in a two-necked flask (50 mL), replace with nitrogen, add ultra-dry tetrahydrofuran (15 mL), stir at 0°C and add potassium tert-butoxide (1.949 mL, 1.949 mmol) in tetrahydrofuran dropwise, and continue stirring at 0°C for 10 minutes. (3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladec-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-11-one 106-4 (260 mg, 0.390 mmol) was dissolved in ultra-dry tetrahydrofuran (5 mL) and added dropwise into the reaction bottle. The reaction mixture was stirred at room temperature for 1 hour. The reaction was quenched with 5% sodium dihydrogen phosphate solution, extracted with ethyl acetate, and the organic phase was purified by silica gel column chromatography (petroleum ether/dichloromethane = 2/1) to obtain (9R)-9-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-tetramethyl-11-methylidene-4,5,5a,5b,6,7,7a,8,9, 10,10a,10b,11,12,12a,12b-Hexadecahydro-3aH-cyclopenta[1',2':1,2]phenanthro[ ^7,8 -d][1,3]dioxolan-8-yl]-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecane 106-5 (210 mg, 0.253 mmol, 64.81%) was a colorless transparent oil. NMR (400MHz, CDCl3) δ7.60(d,J＝7.6Hz,1H),7.36–7.28(m,1H),7.08(t,J＝7.4Hz,1H),6.95(d,J＝8.1Hz,1H),5.21(dt,J＝7 .8,4.1Hz,1H),4.85(s,1H),4.73(s,1H),4.21–4.16(m,1H),4.16–4.07(m,1H),3.95(s,3H),2.53(t,J＝12.8Hz,1H),2.24 (d,J＝10.4Hz,2H),2.12(t,J＝14.2Hz,3H),2.04–1.90(m,3H),1.79(ddd,J＝14.0,11.9,3.6Hz,3H),1.72(s,2H),1.66(s,4 H),1.58–1.49(m,J＝9.1,3.9Hz,6H),1.45(s,3H),1.41–1.37(m,2H),1.31(s,3H),1.24(dd,J＝11.4,7.0Hz,2H),1.18–1.11 (m,2H),1.03(s,9H),0.93–0.86(m,1H),0.82(s,3H),0.15(dd,J＝7.9,2.2Hz,4H),-0.00(s,3H).

Step 6: (9R)-9-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-tetramethyl-11-methylidene-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8 -d][1,3]dioxolan-8-yl]-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecane 106-5 (18 mg, 0.027 mmol) was dissolved in tetrahydrofuran (1 mL), hydrochloric acid (2.520 mL, 7.560 mmol) was added, and stirred at 40°C for 3 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1:1), and the raw material was reacted. Water was added to the reaction solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and dried by spin drying. The crude product was purified by silica gel column chromatography (PE/EA = 1/1) to obtain 24-[hydroxy(2-methoxyphenyl)methyl]-7-methylidene-5α-cholane-3β,4β-diol 106 (6 mg, 0.011 mmol, 41.24%) as a white solid.

Compound 106: ¹ H NMR (400 MHz, CDCl3) δ7.30 (d, J = 5.8 Hz, 1H), 7.24 (d, J = 7.8 Hz, 1H), 6.97 (dd, J = 15.5, 8.1 Hz, 1H), 6.88 (d, J = 8.3 Hz, 1H), 4.91–4.80 (m, 1H), 4.65 (d, J = 42.3 Hz, 2H), 3.86 (s, 3H), 3.79 (s, 1H), 3.61–3.49 (m, 1H), 2.49 (t, J = 12.8 Hz, 1H), 2.09–1.92 (m, 5H), 1.84 (d d,J＝23.5,14.4Hz,2H),1.78–1.66(m,5H),1.40(d,J＝2.7Hz,3H),1.34–1.28(m,2H),1.25(s,2H),1.18(s,1H),1.15(s,3H),1.11 (d,J＝11.3Hz,3H),1.01–0.96(m,1H),0.92(dd,J＝6.4,3.9Hz,3H),0.75(t,J＝9.3Hz,1H),0.67(d,J＝2.5Hz,3H).LC-MS:[M+H-H2O] ⁺ =493.45

Embodiment 109

Compound 109 Preparation of 3β,25-dihydroxy-5α-cholestane-4-carbonitrile

Step 1: In a 50 mL round-bottom flask at room temperature, 3β-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-25-hydroxycholest-6(5)-ene-4-carbaldehyde 23-3 (90 mg, 0.17 mmol) was dissolved in acetonitrile (6 mL), and sodium acetate (230 mg, 2.8 mmol), hydroxylamine hydrochloride (115 mg, 1.7 mmol) and water (2 ml) were added at room temperature, followed by stirring at room temperature for 30 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 4:1). After the reaction was completed, the mixture was quenched with water (20 mL), extracted with ethyl acetate (20 mL × 3), and the organic phase was washed with saturated brine (30 mL). The reaction mixture was washed with water and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=4:1) to give a white solid [(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methanealdehyde oxime 109-1 (53 mg, purity: 90%, yield: 51.58%). ¹ H NMR (400MHz, CDCl ₃ )δ＝7.59(d,J＝9.2,1H),3.75(dd,J＝15.4,5.9,1H),2.61–2.54(m,1H),2.01–1.27(m,25H),1.21(s,6H ),1.11–1.00(m,5H),0.91(d,J＝6.5,3H),0.85(s,9H),0.79(s,3H),0.62(s,3H),0.03(d,J＝3.0,6H).

Step 2: In a 50 mL round-bottom flask at room temperature, [(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methanealdehyde oxime 109-1 (43 mg, 0.077 mmol) was dissolved in tetrahydrofuran (2.5 mL), and diphenyl disulfide (67 mL) was added at room temperature. g, 0.31mmol), tributylphosphine (62mg, 0.31mmol), then stirred at room temperature for 6 hours, TLC (petroleum ether: ethyl acetate = 8: 1) monitored the reaction, after the reaction was completed, quenched with water (10mL), extracted with ethyl acetate (10mL×3), the organic phase was washed with saturated brine (20mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8: 1) to give a white solid 3β-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-25-hydroxycholest-4-carbonitrile 109-2 (31mg, purity: 90%, yield: 67.04%). ¹ H NMR (400MHz, CDCl ₃ )δ＝3.61–3.50(m,1H),2.69(t,J＝4.7,1H),1.95–1.21(m,26H),1.14(s,6H),1. 01(s,3H),0.98–0.88(m,3H),0.84(d,J＝5.0,12H),0.58(s,3H),-0.00(s,6H). ¹³ C NMR (101MHz, CDCl ₃ )δ142.57,119.68,71.12,70.74,56.45,56.12,55.12,45.76,44.41,42.55,40.84,39.80,36.60,36.41,36.14,35.7 1,35.19,31.90,31.52,29.71,29.40,29.20,29.07,28.22,27.08,25.71,24.17,20.72,18.64,18.02,13.09,12.05.

Step 3: In a 50 mL round-bottom flask at room temperature, 3β-{[dimethyl(2-methylprop-2-yl)methylsilyl]oxy}-25-hydroxycholestane-4-carbonitrile 109-2 (31 mg, 0.057 mmol) was dissolved in a 1 mol tetrabutylammonium fluoride tetrahydrofuran solution (2 mL), stirred at room temperature for 2 hours, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 4:1). After the reaction was completed, it was quenched with water (10 mL), extracted with ethyl acetate (10 mL×3), and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 4:1) to obtain a white solid 3β,25-dihydroxy-5α-cholestane-4-carbonitrile 109 (18.02 mg, purity: 100%, yield: 73.59%). Compound 109: ¹ H NMR (400 MHz, CDCl ₃ )δ＝3.73–3.63(m,1H),2.95(t,J＝4.3,1H),2.03–1.25(m,26H),1.21(s ,6H),1.09(s,3H),1.07–0.95(m,4H),0.91(d,J=6.5,3H),0.65(s,3H). ¹³ C NMR (101MHz, CDCl ₃ )δ119.61,71.13,70.24,56.38,56.12,54.98,45.71,44.40,42.54,40.17,39.73,36.66,36.40,36.13 ,35.71,35.17,31.83,29.71,29.39,29.20,28.71,28.21,27.11,24.16,20.74,18.64,13.06,12.05.. LC-MS: [M+HH ₂ O] ⁺ =412.5.

Embodiment 114

Compound 114 Preparation of 24-[Hydroxy(2-methoxyphenyl)methyl]-7-(hydroxymethyl)-5α-cholane-3β,4β-diol

Step 1: (9R)-9-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-tetramethyl-11-methylidene-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1 ,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecane 106-5 (180 mg, 0.271 mmol) was dissolved in ultra-dry tetrahydrofuran (5 mL), replaced with nitrogen, and borane dimethyl sulfide (0.406 mL, 0.812 mmol) was added under an ice-water bath. The mixed solution was stirred at room temperature for 2 hours. Then, aqueous sodium hydroxide solution (0.162 mL, 1.624 mmol) and hydrogen peroxide (184.09 mg, 1.624 mmol) were added dropwise to the reaction solution. The mixed solution was stirred at room temperature overnight. The reaction solution was quenched with aqueous sodium bisulfite solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and spin-dried. The crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 3/1) to obtain [(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-11-yl]methanol 114-1 ( ⁹⁰ mg, 0.112 mmol, 41.38%) was obtained as a white solid. NMR(400MHz,cdcl3)δ7.60(d,J＝7.6Hz,1H),7.36–7.29(m,1H),7.09(d,J＝6.9Hz,1H),6.95(d,J＝8.2Hz,1H),5.21(dt,J＝7.6,3.9Hz,1H),4.14(s,2 H),3.96(d,J＝3.3Hz,3H),3.90(s,1H),3.83(s,1H),2.05(s,2H),1.93(dt ,J＝8.5,4.3Hz,5H),1.87–1.83(m,1H),1.81(d,J＝2.3Hz,1H),1.75(dd,J＝ 13.1,3.3Hz,3H),1.71–1.67(m,2H),1.66(s,4H),1.53(ddd,J＝15.3,9.1, 3.3Hz,5H),1.44(s,3H),1.42(d,J＝2.8Hz,1H),1.41–1.38(m,3H),1.23(s ,3H),1.18(d,J＝9.5Hz,2H),1.03(d,J＝2.9Hz,9H),1.01(d,J＝2.2Hz,2H), 0.98–0.91(m,1H),0.78(s,3H),0.19–0.11(m,4H),0.00(d,J＝0.9Hz,3H).

Step 2: [(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-silanadecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a ,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-11-yl]methanol 114-1 (20 mg, 0.029 mmol) was dissolved in tetrahydrofuran (1 mL), hydrochloric acid (2 mL, 6.000 mmol) was added, and the mixture was stirred at room temperature for 2 hours. Water was added to the reaction solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and dried by rotation. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:4) to obtain 24-[hydroxy(2-methoxyphenyl)methyl]-7-(hydroxymethyl)-5α-cholane-3β,4β-diol as a white solid 114 (4 mg, 0.007 mmol, 23.26%)

Compound 114: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.27 (s, 1H), 7.25 (s, 1H), 6.96 (s, 1H), 6.89 (d, J = 8.2 Hz, 1H), 4.83 (d, J = 7.0 Hz, 1H), 3.86 (s, 3H), 3.77 (d, J = 6.9 Hz, 1H), 3.73 (s, 1H), 3.67 (t, J = 10.6 Hz, 1H), 3.58 (d, J = 10.9 Hz, 1H), 1.88 (d, J = 12.5 Hz, 5H) ,1.76–1.63(m,7H),1.39(dd,J＝13.8,7.3Hz,3H),1.34–1.24(m,5H),1.07(s,3H),1.00(d,J＝13.9Hz,3 H),0.93–0.87(m,3H),0.81(dd,J＝13.8,9.3Hz,2H),0.63(d,J＝2.3Hz,3H),0.07(s,2H).LC-MS:[M+H-2H ₂ O] ⁺ = 493.45.

Embodiment 118

Preparation of Compound 118 4-(Hydroxymethyl)-5α-cholest-3β,25-diol

Step 1: Dissolve compound (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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-8 (150 mg, 0.326 mmol, 1.0 eq) in ethyl acetate (5 mL), add platinum dioxide (36.97 mg, 0.163 mmol, 0.5 eq), and stir the resulting mixture in H ₂ at 25° C. for 3 h. The reaction progress was monitored by TLC (petroleum ether:ethyl acetate=3:1), and the raw material was completely consumed. After filtration, the filtrate was concentrated to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate: 80:20) to obtain a white solid (5R)-5-[(1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-7-acetoxy-6-hydroxy-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester IV-1 (100 mg, 0.195 mmol, 59.74%). ¹ H NMR (400MHz, CDCl3) δ4.72(m,1H),3.83(s,1H),3.66(s,3H),2.27(dd,J＝16.4,8.7Hz,2H),2.10(d,J＝8. 1Hz,3H),1.79(dddd,J＝37.0,16.1,10.1,3.7Hz,8H),1.29(m,8H),0.99(m,14H),0.62(d,J＝14.3Hz,4H).

Step 2: In a 50 mL round-bottom flask, (5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester IV-1 (200 mg, 0.43 mmol) was dissolved in dichloromethane (10 mL), and N-methylmorpholine oxide (76 mg, 0.65 mmol), tetrapropylammonium perruthenate (30 mg, 0.086 mmol) and tetraA molecular sieves (190 mg, 0.43 mmol) were added at room temperature, and the mixture was stirred at room temperature for 24 hours. The reaction was monitored by TLC (petroleum ether/ethyl acetate = 10/1). After the reaction, water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL × 3). The extracted organic solution was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified and separated by silica gel column chromatography (eluent: petroleum ether/ethyl acetate = 10/1) to obtain a white solid (5R)-5-[(1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-7-acetoxy-9a, 11a-dimethyl-6-oxyylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 118-1 (170 mg, purity: 90%, yield: 76.83%). ¹ H NMR(400MHz, CDCl3)δ5.16(dd,J＝12.0,7.5Hz,1H),3.67(s,3H),2.31–2.18(m,4H),2.1 5(d,J＝3.2Hz,3H),2.01–1.00(m,25H),0.92(d,J＝6.6Hz,3H),0.75(s,3H),0.65(s,3H). ^13C NMR(101MHz, CDCl3)δ=205.71,174.32,170.18,76.20,57.39,56.06,55.90,54.24,51.44,42.67,42.55,39.80,3 5.93,35.48,35.36,34.91,34.50,30.23,28.50,28.12,24.10,21.70,21.52,20.75,20.17,18.54,13.69,12.03.

Step 3: In a 50 mL round-bottom flask, methyl triphenylphosphine bromide (388 mg, 1.09 mmol) was suspended in tetrahydrofuran (4 mL) at room temperature, and a 1 M/L solution of potassium tert-butoxide in tetrahydrofuran (1.09 mL, 1.09 mmol) was added dropwise at 0°C. After the mixture was stirred at room temperature for 2 hours, a solution of (5R)-5-[(1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-7-acetoxy-9a, 11a-dimethyl-6-oxyylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 118-1 (100 mg, 0.22 mmol) in tetrahydrofuran (1 mL) was added dropwise at 0°C, and the mixture was stirred at room temperature for 5 hours. The reaction was monitored by TLC (petroleum ether/ethyl acetate = 10/1). After the reaction, water (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL × 3). The extracted organic solution was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified and separated by silica gel column chromatography (eluent: petroleum ether/ethyl acetate = 10/1) to obtain a white solid (5R)-5-[(1R, 3aS, 3bS, 5aR, 7S, 9aR, 9bS, 11aR)-7-acetoxy-9a, 11a-dimethyl-6-methylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 118-2 (64 mg, purity: 90%, yield: 57.85%). ¹ H NMR(400MHz, CDCl3)δ5.20(d,J＝11.1Hz,1H),4.69(s,1H),4.59(s,1H),3.66(s,3H),2.31–2.25(m,3H),2.13(s, 3H),2.06–1.64(m,9H),1.60–1.29(m,13H),1.12–1.02(m,5H),0.96(s,3H),0.91(d,J＝6.5Hz,3H),0.65(s,3H). ^13C NMR(101MHz, CDCl3)δ=174.35,170.56,147.24,129.65,115.27,103.83,74.35,56.35,55.90,54.43,52.43,51.44,42.52,39 .90,39.43,37.97,35.49,35.39,34.99,34.53,31.34,30.49,28.16,24.12,23.47,21.53,21.00,20.90,18.55,13.22,12.05.

Step 4: In a 50 mL round-bottom flask at room temperature, (5R)-5-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-acetoxy-9a,11a-dimethyl-6-methylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester 118-2 (68 mg, 0.15 mmol) was dissolved in anhydrous tetrahydrofuran (4 mL). The reaction was cooled to 0°C under nitrogen protection, and 3 mol/L methyl magnesium bromide tetrahydrofuran solution (0.50 mL, 1.50mmol), then stirred at room temperature for 2 hours, monitored by TLC (petroleum ether: ethyl acetate = 4: 1), after the reaction, the system was cooled to 0 ° C, quenched with saturated aqueous ammonium chloride solution (15mL), extracted with ethyl acetate (10mL×3), the organic phase was washed with saturated brine (20mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1) to obtain a white solid 4-methylidene cholesteryl-3β,25-diol 118-3 (34mg, purity: 90%, yield: 46.46%). ¹ H NMR (400MHz, CDCl3) δ＝4.83(dd,J＝115.4,50.6,2H),4.07–3.81(m,1H),2.27(dt,J＝16.4,8.4,1H),1.97(dd,J＝10.2,3 .8,2H),1.85–1.72(m,4H),1.63–1.32(m,20H),1.26(s,3H),1.21(s,6H),1.12–1.00(m,6H),0.91(s,3H),0.66(s,3H). ^13C NMR (101MHz, CDCl3) δ152.30,151.31,102.19,101.34,72.35,71.24,70.11,55.38,55.24,55.2 0,55.13,54.36,53.48,53.45,48.86,43.40,41.58,41.50,39.04,38.94,38.63,37.58,36.75,3 6.23,35.39,34.72,34.22,33.99,32.02,30.56,29.46,28.68,28.34,28.32,28.19,27.25,23. 23,23.18,23.13,22.44,21.67,20.69,19.98,19.75,17.61,13.11,12.30,11.91,11.05,11.04.

Step 5: In a 50 mL round-bottom flask, 4-methylidene cholesteryl-3β,25-diol 118-3 (34 mg, 0.082 mmol) was dissolved in tetrahydrofuran (4 mL) at room temperature, and a tetrahydrofuran solution of 2 mol/L borane dimethyl sulfide complex (0.12 mL, 0.24 mmol) was added dropwise at 0°C. After the mixture was stirred at room temperature for 3 hours, 10 mol/L sodium hydroxide aqueous solution (0.049 mL, 0.49 mmol) and 30% hydrogen peroxide aqueous solution (56 mg, 0.49 mmol) were added dropwise to the reaction solution at 0°C, and the mixture was stirred at room temperature for 16 hours. TLC (petroleum ether/ethyl acetate = 2/1) was used to monitor the reaction. After the reaction was completed, water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL×3). The extracted organic solution was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified and separated by silica gel column chromatography (eluent: petroleum ether/ethyl acetate = 2/1) to obtain a white solid 4-(hydroxymethyl)-5α-cholest-3β,25-diol 118 (22.27 mg, purity: 73.65%, yield: 46.05%). ¹ H NMR (400 MHz, CDCl3) δ = 4.26 (t, J = 10.3, 1H), 3.84 (dd, J = 11.2, 5.1, 1H), 3.60-3.53 (m, 1H), 2.35-2.27 (m, 1H), 2.05-1.66 (m, 15H), 1.59-1.33 (m, 16H), 1.21 (s, 6H), 0.91 (d, J = 6.5, 3H), 0.81 (s, 3H), 0.65 (s, 3H). ¹³ C NMR(101MHz, DMSO)δ=99.99,68.64,66.35,59.50,51.76,51.45,49.90,42.50,39.68,37.77,36.43,35.21,33.06,31.69 ,31.01,30.32,28.83,26.77,24.96,24.60,24.47,23.51,19.38,18.86,17.78,16.05,13.89,8.16,7.33.LC-MS: [M+H-2H ₂ O] ⁺ =399

Example 13

Compound 13 Preparation of Acetic Acid-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-Hydroxy-1-[(2R)-6-Hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methyl Ester and Acetic Acid-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-Acetoxy-1-[(2R)-6-Hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methyl Ester

Step 1: 4-(Hydroxymethyl)-5α-cholest-3β,25-diol 118 (180 mg, 0.41 mmol) was dissolved in dichloromethane (2 mL) and tetrahydrofuran (2 mL) in a 50 mL round-bottom flask at room temperature. Triethylamine (126 mg, 1.24 mmol), 4-dimethylaminopyridine (5 mg, 0.041 mmol) and acetic anhydride (42 mg, 0.41 mmol) were added at room temperature, followed by stirring at room temperature for 1 hour. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 3:1). After the reaction was completed, the reaction was quenched with water (30 mL), extracted with ethyl acetate (20 mL×3), the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1) to obtain white solid acetic acid-[(1R, 3aS, 3bS, 5aS, 7S, 9aR, 9bS, 11aR)-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methyl ester 13 (180 mg, purity: 90%, yield: 82.06%).

Compound 13: ¹ H NMR (400 MHz, CDCl3) δ = 4.34–4.18 (m, 2H), 3.81–3.72 (m, 1H), 2.04 (s, 3H), 1.99– 1.93(m,1H),1.82–1.69(m,4H),1.68–1.51(m,9H),1.44–1.33(m,8H),1.29–1.24(m ,4H),1.21(s,6H),1.09–0.95(m,5H),0.91(d,J＝6.5,3H),0.73(s,3H),0.63(s,3H). ^13C NMR (101MHz, CDCl3)δ=171.05,73.11,71.12,61.73,56.46,56.14,55.32,47.67,46.37,44.42,42.51,39.84,37.20 ,36.42,35.73,35.55,32.54,29.37,29.20,28.23,26.91,26.76,24.23,21.25,20.89,20.77,18.63,14.03,12.01.

Embodiment 119

Preparation of Compound 119 4β-Hydroxy-3β-Hydroxy-24-[Hydroxy(2-methoxyphenyl)methyl]-5α-cholane-7-carboxylic acid

Step 1: [(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-silanadecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12 a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-11-yl]methanol 114-1 (70 mg, 0.102 mmol) was dissolved in dichloromethane 99.9% (3 mL), and N-methylmorpholine oxide (21 mg, 0.179 mmol), tetrapropylammonium perruthenate (14 mg, 0.040 mmol) and molecular sieves were added. The mixture was stirred at room temperature for 30 minutes. Water was added, extracted with ethyl acetate, and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate and spin-dried. The crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 4/1) to give (3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladec-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-11-carbaldehyde 119-1 (60 mg, 0.075 mmol, 73.08%) as a white solid. ¹ H NMR (400MHz, CDCl ₃ )δ9.95(s,1H),7.45(d,J＝7.7Hz,1H),7.18(s,1H),6.94(s,1H),6.81(d,J ＝8.2Hz,1H),5.07(d,J＝3.6Hz,1H),4.00(s,2H),3.81(d,J＝3.4Hz,3H),2.5 9(s,1H),1.97(d,J＝12.8Hz,1H),1.85(d,J＝5.4Hz,6H),1.63(d,J＝18.7Hz ,5H),1.50(s,3H),1.43(s,3H),1.35(d,J＝15.3Hz,3H),1.30(s,3H),1.27– 1.19(m,3H),1.18–1.10(m,3H),1.07(s,3H),1.04(s,2H),0.97(d,J＝6.6Hz, 1H), 0.89 (s, 9H), 0.67 (s, 2H), 0.02 (t, J = 2.7Hz, 3H), 0.00 (d, J = 3.3Hz, 6H).

Step 2: (3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-silanadecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10 b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolane-11-carbaldehyde 119-1 (55 mg, 0.081 mmol) was dissolved in tert-butyl alcohol (1 mL), 2-methyl-2-butene (33.98 mg, 0.485 mmol) was added under ice-water bath, and stirred for 1 minute. Then, sodium chlorite (25.56 mg, 0.283 mmol) aqueous solution and sodium dihydrogen phosphate (2.62 mg, 0.022 mmol) aqueous solution were added. The mixture was moved to room temperature and stirred for 1 hour. Water was added to the reaction solution, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride solution and dried with anhydrous sodium sulfate, and then spin-dried. The crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 3/1) to give (3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolane-11-carboxylic acid 119-2 (50 mg, 0.062 mmol, 76.39%) as a white solid. ¹ H NMR (400MHz, CDCl ₃ )δ7.45(d,J＝7.6Hz,1H),7.17(t,J＝7.7Hz,1H),6.93(t,J＝7.5Hz,1H),6.80(d,J＝8.2Hz,1H),5.06(dd,J＝7.1,3.0Hz,1H),3.9 9(ddd,J＝14.2,10.0,5.0Hz,2H),3.80(s,3H),2.79(s,1H),2.09(dd,J＝13.4,5.7Hz,1H),1.93(d,J＝11.3Hz,1H),1.83(dd,J＝1 3.9,11.3Hz,4H),1.69(dd,J＝28.9,9.6Hz,4H),1.63–1.53(m,8H),1.47(s,2H),1.37(dd,J＝21.0,11.0Hz,4H),1.29(s,3H),1. 19(d,J＝15.1Hz,2H),1.16–1.08(m,3H),1.06(s,3H),1.03–0.92(m,3H),0.88(s,9H),0.64(s,3H),0.01(s,3H),-0.15(s,3H).

Step 3: Dissolve (3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolane-11-carboxylic acid 119-2 (50 mg, 0.072 mmol) in tetrahydrofuran (2 mL) and add hydrochloric acid (1 mL, 3.000 mmol). The mixture was stirred at 40°C for 2.5 hours. Water was added to the reaction solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and spin-dried. The crude product was purified by flash chromatography (petroleum ether/ethyl acetate = 90%-100%) to obtain 4β-hydroxy-3β-hydroxy-24-[hydroxy(2-methoxyphenyl)methyl]-5α-cholane-7-carboxylic acid 119 (11.5 mg, 0.015 mmol, 20.68%) as a white solid.

Compound 119: ¹ H NMR (400MHz, DMSO) δ11.71(s,1H),7.38(d,J＝7.6Hz,1H),7.17(t,J＝7.2Hz,1H),6.92(dd,J＝7.5,3.9Hz, 2H),4.85(s,2H),4.33(d,J＝ 5.8Hz,1H),3.98(d,J＝2.7Hz,1H),3.75(s,3H),3.40(s,1H),3.28(s,1H),2.54(s,1H),2.02(td, J＝13.3,5.9Hz,1H),1.87(d,J＝11.1H z,1H),1.78–1.68(m,1H),1.58(dd,J＝11.4,5.7Hz,2H),1.48(s,3H),1.40(dd,J＝22.5,13.8Hz,8H),1.32 –1.25(m,2H),1.23(s,1H), 1.21–1.11(m,2H),1.09–0.97(m,4H),0.96(s,3H),0.90(d,J＝13.1Hz,1H),0.84(d,J＝6.2Hz,3H),0.59 (d,J=2.3Hz,3H).LC-MS: [M+HH ₂ O] ⁺ =525.40.

Embodiment 121&122&125

Compound 121: [(1R,3aS,3bS,5aS,7S,9aS,9bS,11aR)-4,4-difluoro-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]- 9a,11a-Dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]acetonitrile,

Compound 122: Preparation of (1R, 3aS, 3bS, 5aR, 7S, 9aR, 9bS, 11aR)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethyl-6-methylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol and Compound 125: Preparation of (1R, 3aS, 3bS, 5aS, 7S, 9aS, 9bS, 11aR)-4,4-difluoro-6-(hydroxymethyl)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol

Step 1: In a 50 mL round-bottom flask at room temperature, (1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-7-(benzyloxy)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-ol 79-7A (790 mg, 1.445 mmol) was dissolved in 1,2-dichloroethane (60 mL), and Dess-Martin periodinane (1838.04 mg, 4.335 mmol) was added at room temperature, and stirred at room temperature for 2 hours. The reaction was monitored by TLC (ethyl acetate/petroleum ether = 4:1). After the reaction, saturated aqueous sodium sulfite solution (30 mL) and saturated aqueous sodium bicarbonate solution (30 mL) were added to quench the reaction, and the mixture was extracted with dichloromethane (50 mL × 3). The organic phase was washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain a white solid. (1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-(Benzyloxy)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-one 121-1 (750 mg, purity: 90%, yield: 85.76%). ¹ H NMR (400MHz, CDCl3) δ = 8.09 (d, J = 7.5, 2H), 7.58 (t, J = 7.4, 1H), 7.45 (t, J = 7.7, 2H), 5.44 (dd, J = 12.0, 7.4, 1H), 2.63 (d,J＝12.3,1H),2.47–2.37(m,1H),2.23–1.23(m,25H),1.22(s,6H),0.94(d,J＝6.5,3H),0.85(s,3H),0.68(s,3H). ¹⁹ F NMR (376MHz, CDCl ₃ ) δ = -90.57 (d, J = 240.1, 1F), -111.72 (d, J = 240.0, 1F).

Step 2: In a 50 mL round-bottom flask, methyltriphenylphosphine bromide (1.3 g, 3.67 mmol) was dissolved in tetrahydrofuran (20 mL) at room temperature, and 1 M/L potassium tert-butoxide tetrahydrofuran solution (3.67 mL, 3.67 mmol) was added dropwise at 0°C, and the mixture was stirred at room temperature for 2 hours. A solution of (1R, 3aS, 3bS, 5aR, 7S, 9aR, 9bS, 11aR)-7-(benzyloxy)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-one 121-1 (400 mg, 0.73 mmol) in tetrahydrofuran (5 mL) was added dropwise to the reaction solution at room temperature, and the mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC (petroleum ether/ethyl acetate = 8/1). After the reaction was completed, water (30 mL) was added to the reaction solution, extracted with ethyl acetate (20 mL × 3), washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified and separated by silica gel column chromatography (petroleum ether/ethyl acetate = 8/1) to obtain a white solid (6R)-6-[(1R, 3aS, 3bS, 5aR, 7S, 9aR, 9bS, 11aR)-7-(benzyloxy)-4,4-difluoro-9a, 11a-dimethyl-6-methylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylheptan-2-ol 121-2 (158 mg, purity: 90%, yield: 35.68%). ¹ H NMR (400MHz, CDCl3) δ＝8.18–8.06(m,2H),7.59(q,J＝7.4,1H),7.48(t,J＝7.7,2H),5.47(d,J＝11.3,1H),4.77(s,1H ),4.73(s,1H),2.45–2.34(m,2H),2.18–1.34(m,25H),1.21(s,6H),1.06(s,3H),0.93(d,J＝6.5,3H),0.68(s,3H). ¹⁹ F NMR (376MHz, CDCl3) δ = -89.23 (d, J = 238.5, 1F), -110.87 (d, J = 238.4, 1F).

Step 3: In a 50 mL round-bottom flask at room temperature, (6R)-6-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-(benzyloxy)-4,4-difluoro-9a,11a-dimethyl-6-methylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylheptan-2-ol 121-2 (158 mg, 0.29 mmol) was dissolved in tetrahydrofuran (2 mL) and methanol (2 mL), and lithium hydroxide, monohydrate (73 mg, 1.74 mmol) and water (1 mL) were added at room temperature, followed by stirring at room temperature for 2 hours. TLC (petroleum ether:ethyl acetate) was used for the analysis. =5:1 to monitor the reaction. After the reaction was completed, the mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL×3), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1) to obtain a white solid (1R, 3aS, 3bS, 5aR, 7S, 9aR, 9bS, 11aR)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethyl-6-methylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol 122 (120 mg, purity: 95%, yield: 86.52%).

Compound 122: ¹ H NMR (400 MHz, CDCl3) δ = 4.93 (d, J = 1.2, 1H), 4.79 (d, J = 1.4, 1H), 3.91 (d, J = 11.0, 1H), 2.54–2.44 (m, 1H), 2.38–2.20 (m, 2H), 2.08–1.24 (m, 25H), 1.21 (s, 6H), 0.95 (s, 3H), 0.93 (d, J = 6.5, 3H), 0.68 (s, 3H). ¹³ C NMR(101MHz, CDCl3)δ=151.10,104.32,71.25,71.13,55.20,50.57,50.22,48.51,44.39,43.05,41.65,3 9.31,38.82,37.29,36.40,35.70,30.17,29.31,29.24,28.49,25.37,20.95,20.75,18.73,12.37,11.87. ¹⁹ F NMR (376MHz, CDCl3) δ = -89.00 (d, J = 237.7, 1F), -110.95 (d, J = 237.6, 1F). LC-MS: [M+H-2H ₂ O] ⁺ =417.35.

Step 4: In a 50 mL round-bottom flask, (1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-4,4-difluoro-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-6-methylidene hexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol 122 (213 mg, 0.47 mmol) was dissolved in tetrahydrofuran (12 mL) at room temperature, and dihydrogen sulfoxide was added dropwise at 0 °C. Borane dimethyl sulfide complex tetrahydrofuran solution (0.71mL, 1.41mmol), after the mixture was stirred at room temperature for 3 hours, ten moles of sodium hydroxide aqueous solution (0.28mL, 2.82mmol) and 30% hydrogen peroxide aqueous solution (320mg, 2.82mmol) were added dropwise to the reaction solution at 0°C, and the mixture was stirred at room temperature for 16 hours. TLC (petroleum ether/ethyl acetate = 2/1) monitored the reaction. After the reaction was completed, water (20mL) was added to the reaction solution, extracted with ethyl acetate (20mL×3), and the extracted organic solution was washed with saturated brine (30mL), dried with anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified and separated by silica gel column chromatography (eluent: petroleum ether/ethyl acetate = 2/1) to give a white solid (1R, 3aS, 3bS, 5aS, 7S, 9aS, 9bS, 11aR)-4,4-difluoro-6-(hydroxymethyl)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol 125 (184 mg, purity: 95%, yield: 78.92%). Compound 125: ¹ H NMR (400 MHz, CDCl3) δ = 4.25 (t, J = 10.6, 1H), 3.82 (dd, J = 11.1, 5.3, 1H), 3.63 (dd, J = 10.9, 4.2, 1H), 2.47-2.33 (m, 2H), 2.06-1.93 (m, 2H), 1.88-1.39 (m, 21H), 1.21 (s, 6H), 1.16-1.01 (m, 5H), 0.93 (d, J = 6.5, 3H), 0.85 (s, 3H), 0.67 (s, 3H). ¹⁹ F NMR (376MHz, CDCl3) δ = -88.57 (d, J = 237.7, 1F), -110.59 (d, J = 237.6, 1F). ¹³ C NMR (101MHz, CDCl3) δ72.09,71.11,69.65,66.63,65.39,64.38,64.05,63.39,56.85,55.25,53.89,50.87,48.88,46. 80,44.39,43.03,40.85,39.27,35.71,30.18,29.73,29.34,29.25,28.50,23.39,20.76,18.73,11.86.LC-MS: [M+Na] ⁺ =493.3,

Step 5: In a 50 mL round-bottom flask at room temperature, (1R,3aS,3bS,5aS,7S,9aS,9bS,11aR)-4,4-difluoro-6-(hydroxymethyl)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-7-ol 125 (159 mg, 0.34 mmol) was dissolved in 1,2-dichloroethane (8 mL), and triethylamine (205 mg, 2.04 mmol), 4-dimethylaminopyridine (41 mg, 0.34 mmol) and p-toluenesulfonyl chloride (129 mg, 0.68 mmol) were added at room temperature, followed by stirring at 50 °C for 16 hours, and the reaction was monitored by TLC (petroleum ether:ethyl acetate=3:1). After the reaction was completed, the reaction was quenched with water (20 mL), extracted with ethyl acetate (20 mL×3), the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain a white solid 4-methylbenzenesulfonic acid-[(1R, 3aS, 3bS, 5aS, 7S, 9aS, 9bS, 11aR)-4,4-difluoro-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methyl ester 121-3 (155 mg, purity: 90%, yield: 66.09%). ¹ H NMR (400MHz, CDCl3) δ = 7.80 (d, J = 8.3, 2H), 7.36 (d, J = 8.1, 2H), 4.29 (dd, J = 9.8, 4.2, 1H), 4.16 (t, J = 9.8, 1H), 3.72 (dd, J = 11.8, 5.1, 1 H),2.45(s,3H),2.37–2.19(m,2H),2.02–1.33(m,22H),1.22(s,6H),1.16–0.99(m,5H),0.93(d,J＝6.5,3H),0.83(s,3H),0.65(s,3H). ^19F NMR (376MHz, CDCl3) δ = -88.75 (d, J = 238.4, 1F), -110.46 (d, J = 238.3, 1F).

Step 6: In a 50 mL round-bottom flask at room temperature, 4-methylbenzenesulfonic acid-[(1R,3aS,7S,9aS,11aR)-4,4-difluoro-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]methyl ester 121-3 (45 mg, 0.072 mmol) was dissolved in N,N-dimethylformamide (2 mL), sodium cyanide (18 mg, 0.36 mmol) was added at room temperature, and then heated at 100 ° C. The mixture was stirred for 16 hours and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 4:1). After the reaction was completed, the mixture was quenched with water (10 mL), extracted with ethyl acetate (10 mL×3), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 4:1) to obtain a white solid [(1R, 3aS, 3bS, 5aS, 7S, 9aS, 9bS, 11aR)-4,4-difluoro-7-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-yl]acetonitrile 121 (7.27 mg, purity: 100%, yield: 21.04%).

Compound 121: ^1H NMR (400MHz, CDCl3) δ = 4.93 (s, 0.5H), 4.79 (s, 0.5H), 4.53 (dd, J = 8.6, 1.6, 0.5H), 4.28 (t, J = 8.0, 0.5H), 3.91(d, J＝10.5,0.5H),3.37–3.29(m,0.5H),2.55–1.33(m,29H),1.21(d,J＝1.1,6H),0.98–0.90(m,4.5H),0.68( s,4.5H). ^19F NMR (376MHz, CDCl3) δ = -88.87 (d, J = 136.8, 0.5F), -89.50 (d, J = 136.6, 0.5F), -110.96 (d, J = 237.7, 0.5F), -112.82 ( d,J＝237.5,0.5F). ¹³ C NMR (101MHz, CDCl3)δ=104.31,84.94,71.11,55.20,50.67,48.68,44.40,43.08,43.05,40.40,39.42,39.31,38.82,36.40,35.72,33.50,31.61 ,30.17,29.32,29.25,28.49,25.48,25.44,24.39,21.92,20.98,20.96 ,20.87,20.77,20.75,19.07,18.98,18.73,12.37,12.05,11.90,11.87.

Embodiment 123

Compound 123: Preparation of 4-methylidene-5α-cholest-3β,25-diol

Step 1: 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-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]hexanoic acid methyl ester I-7 (500 mg, 1.124 mmol, 1.0eq) was dissolved in tetrahydrofuran (40mL). After the reaction system was cooled to 0°C, methylmagnesium chloride (3.75mL, 11.24mmol, 3mol/L, 10eq) was slowly added and the reaction system was stirred at room temperature for 2h. The reaction was monitored by TLC plate (petroleum ether: ethyl acetate = 5:1). When the raw material was consumed, the reaction was stopped. 20mL of water was added to the reaction system, extracted with ethyl acetate (50mL×3), the organic phase was collected, dried with anhydrous sodium sulfate, and the organic phase was vacuum-dried to obtain a crude product. The crude product was dissolved in ethyl acetate and subjected to column chromatography (petroleum ether: ethyl acetate = 80:20) to obtain cholest-6(5)-ene-3β,4β,19,25-tetrol 123-1 (450mg, purity: 90%, yield: 89.45%). ¹ H NMR (400MHz, CDCl ₃ )δ5.38–5.32(m,1H),3.51(dd,J＝11.6,6.8Hz,1H),2.33–2.19(m,2H), 2.06–1.92(m,2H),1.88–1.78(m,3H),1.63–1.46(m,6H),1.42–1.31(m, 4H), 1.27 (dd, J = 7.9, 4.1Hz, 2H), 1.21 (s, 6H), 1.08 (ddd, J = 16.7, 13.5, 6.3Hz, 5H), 1.01 (s, 3H), 0.93 (d, J = 6.6Hz, 3H), 0.66 (d, J = 12.7Hz, 3H).

Step 2: Cholest-6(5)-ene-3β,4β,19,25-tetrol 123-1 (450 mg, 1.118 mmol, 1.0 eq) was dissolved in dichloromethane (20 mL), and acetic anhydride (0.315 mL, 3.354 mmol, 3.0 eq), 4-dimethylaminopyridine (27.32 mg, 0.224 mmol, 0.2 eq) and triethylamine (0.777 mL, 5.590 mmol, 5.0 eq) were added in sequence, and stirred at room temperature for 3 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 5:1). After the reaction was completed, the reaction solution was quenched with methanol, and the reaction solution was washed once with saturated sodium bicarbonate (10 mL) and water (10 mL), dried over anhydrous sodium sulfate, concentrated, and then concentrated to dryness. Column chromatography (petroleum ether:ethyl acetate=90:10) gave white solid acetate-(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-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 123-2 (450 mg, 0.911 mmol, purity: 90%, yield: 81.49%). ¹ H NMR (400MHz, CDCl ₃ )δ5.38–5.32(m,1H),4.64-4.60(m,1H),2.33–2.19(m,2H),2.06–1.92(m,3H),1.88–1.78(m,3H),1.63–1.46(m,6H),1.42–1.31(m,4H),1 .27(dd,J＝7.9,4.1Hz,2H),1.21(s,6H),1.08(ddd,J＝16.7,13.5,6.3Hz,5H),1.01(s,3H),0.93(d,J＝6.6Hz,3H),0.66(d,J＝12.7Hz,3H).

Step 3: Dissolve (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-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl acetate 123-2 (450 mg, 1.012 mmol, 1.0 eq) in chloroform (20 mL) under nitrogen protection, add selenium dioxide (281 mg, 2.53 mmol, 2.5 eq) and N-methylmorpholine (307.07 mg, 3.036 mmol), heat the reaction to 75 degrees and stir for 48 hours, TLC (petroleum ether: ethyl acetate = 3: 1) The reaction was monitored and the starting material was almost gone. The reaction mixture was cooled to room temperature and quenched with water. The mixture was extracted with dichloromethane three times. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. The crude product was purified by rapid column chromatography (petroleum ether: ethyl acetate = 70: 30) to give white solid 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-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 123-3 (310 mg, 0.606 mmol, purity: 90%, yield: 59.85%). ¹ H NMR (400MHz, CDCl ₃ )δ5.75-5.70(m,1H),4.80-4.70(m,1H),4.28-4.23(m,1H),2.33–2.19(m,2H),2.06–1.92(m,3H),1.88–1.78(m,3H),1.63–1.46(m,6H),1 .42–1.31(m,4H),1.27(dd,J＝7.9,4.1Hz,2H),1.21(s,6H),1.10-1.06(m,5H),1.01(s,3H),0.93(d,J＝6.6Hz,3H),0.66(d,J＝12.7Hz,3H).

Step 4: Compound 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-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 123-3 (40 mg, 0.087 mmol, 1.0 eq) was dissolved in ethyl acetate (25 mL), and then platinum dioxide (30 mg, 0.13 mmol, 1.5 eq) was added and the reaction system was replaced with a hydrogen atmosphere, and the reaction system was stirred at room temperature for 24 h. TLC (petroleum ether: ethyl acetate = 3: 1) was used to monitor the reaction progress, and the raw materials were consumed. The reaction system was filtered, and the organic phase was collected and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 82:18 to 81:19) to give white solid acetate-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 123-4 (35 mg, purity 90%, yield 78.4%). ¹ H NMR (400MHz, CDCl ₃ )δ4.72(dd,J＝8.2,3.9Hz,1H),3.83(s,1H),2.09(s,3H),1.93(dd,J＝25.6,12.8Hz,2H),1.78(t,J＝11.4Hz,4H),1.67(d,J＝12.4Hz,1H),1.46–1 .30(m,9H),1.29–1.24(m,2H),1.21(s,6H),1.14–1.08(m,3H),1.05(s, 4H), 1.02–0.95 (m, 2H), 0.91 (d, J = 6.5Hz, 3H), 0.65 (s, 3H), 0.61 (s, 1H).

Step 5: Compound acetic acid-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 123-4 (25 mg, 0.054 mmol) was dissolved in dichloromethane (5 mL). Under nitrogen protection, tetrapropylammonium perruthenate (5.69 mg, 0.016 mmol), N-methylmorpholine oxide (9.49 mg, 0.081 mmol) and 4A molecular sieves were added in sequence. The mixture was stirred at room temperature and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 3:1). Water (10 mL) was added to the reaction system, and the aqueous layer was extracted with EA (3×25 mL). The ethyl acetate layers were combined and washed with saturated brine (3×10 mL). The ethyl acetate layers were dried over Na ₂ SO ₄ , filtered and concentrated to obtain a crude product. After the reaction was completed, the reaction solution was filtered, the filtrate was washed with water, the organic phase was dried and concentrated, and the crude product was purified by rapid column chromatography (petroleum ether: ethyl acetate = 81:19) to obtain a white solid acetic acid-(1R, 3aS, 3bS, 5aR, 7S, 9aR, 9bS, 11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethyl-6-oxyylidene hexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 123-5 (25 mg, purity 90%, yield 90%). ¹ H NMR (400MHz, CDCl ₃ )δ5.16(dd,J＝12.2,7.4Hz,1H),2.22(dd,J＝19.4,6.6Hz,2H),2.15(s,3H),2.02–1.71(m,5H),1.60(dd,J＝18.1,1 4.5Hz,3H),1.46–1.24(m,11H),1.21(s,6H),1.14–1.00(m,4H),0.92(d,J＝6.5Hz,3H),0.75(s,3H),0.65(s,3H).

Step 6: In a 50 mL round-bottom flask, methyltriphenylphosphonium bromide (4.65 g, 13.02 mmol) was suspended in tetrahydrofuran (50 mL) at room temperature, and 1 M/L potassium tert-butoxide tetrahydrofuran solution (13.02 mL, 13.02 mmol) was added dropwise at 0°C. After the mixture was stirred at room temperature for 2 hours, a solution of acetic acid-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-6-oxoylidenehexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 123-5 (1.2 g, 2.60 mmol) in tetrahydrofuran (10 mL) was added dropwise at room temperature, and the mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC (petroleum ether/ethyl acetate = 10/1). After the reaction was completed, water (50 mL) was added to the reaction solution, extracted with ethyl acetate (40 mL × 3), washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified and separated by silica gel column chromatography (eluent: petroleum ether/ethyl acetate = 10/1) to obtain white solid acetic acid-(1R, 3aS, 3bS, 5aR, 7S, 9aR, 9bS, 11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethyl-6-methylidene hexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 123-6 (612 mg, purity: 95%, yield: 48.66%). ¹ H NMR (400MHz, CDCl3) δ = 5.11 (dd, J = 11.8, 5.3, 1H), 4.86 (s, 1H), 4.59 (s, 1H), 2.12 (s, 3H), 2.04 –1.24(m,24H),1.21(s,6H),1.16–1.00(m,5H),0.92(d,J＝6.6,3H),0.71(s,3H),0.65(s,3H).

Step 7: In a 50 mL round-bottom flask at room temperature, acetic acid-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethyl-6-methylidene hexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 123-6 (700 g, 1.53 mmol) was dissolved in tetrahydrofuran (10 mL) and methanol (10 mL), and lithium hydroxide, monohydrate (385 mg, 9.16 mmol) and water (5 mL) were added at room temperature, followed by stirring at room temperature for 1 hour, and the reaction was monitored by TLC (petroleum ether:ethyl acetate=4:1). After the reaction, the mixture was diluted with water (30 mL), extracted with ethyl acetate (30 mL×3), the organic phase was washed with saturated brine (60 mL), and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give a white solid 4-methylidene-5α-cholest-3β,25-diol 123 (614 mg, purity: 97%, yield: 93.67%).

Compound 123: ¹ H NMR (400 MHz, CDCl ₃ ) δ＝5.04 (s, 1H), 4.63 (s, 1H), 3.99 (dd, J＝11.4, 5.4, 1H), 1.99 (ddd, J＝11.8, 8.5, 3.3, 2H), 1.89–1.27 (m, 23H), 1.21 (s, 6H), 1.14–1.00 (m, 5H), 0.92 (d, J＝6.5, 3H), 0.69 (s, 3H), 0.65 (s, 3H). ¹³ C NMR(101MHz, CDCl3)δ=153.33,102.35,73.37,71.13,56.28,56.24,54.49,49.90,44.42,42.62,40.08,38.61,37.27,36 .43,35.76,35.26,33.05,31.61,29.36,29.21,28.28,24.27,24.21,21.72,20.78,18.63,12.93,12.07.LC-MS: [M+H-2H ₂ O] ⁺ =381.3.

Embodiment 137:

Compound 137: Preparation of 24-[Hydroxy(2-methoxyphenyl)methyl]-7-methyl-5α-cholane-3β,4β-diol

Step 1: (9R)-9-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-tetramethyl-11-methylidene-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolane -8-yl]-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecane 106-5 (25 mg, 0.038 mmol) was dissolved in methanol (5 mL), Pd/C (13 mg, 0.061 mmol) was added, the system was replaced with hydrogen, and the mixture was stirred at 40°C for 1 hour. Filtered through celite overnight and dried in spun-drying to obtain crude product 137-1 (20 mg), which was directly used in the next step.

Step 2: (9R)-9-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a,11-pentamethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2 ':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecane 137-1 (20 mg, 0.030 mmol) was dissolved in tetrahydrofuran (2 mL), hydrochloric acid (1 mL, 3.000 mmol) was added, and the mixture was reacted at 40 ° C for 2 hours. Water was added to the reaction solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and spin-dried. The crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/1) to obtain 24-[hydroxy(2-methoxyphenyl)methyl]-7-methyl-5α-cholane-3β,4β-diol as a white solid 137 (4.5 mg, 0.007 mmol, 17.32%, yield of two steps).

Compound 137: ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.32–7.27 (m, 1H), 7.25–7.21 (m, 1H), 6.95 (t, J=7.4 Hz, 1H), 6.88 (d, J=8.2 Hz, 1H), 4.85 (dt, J=9.6, 5.8 Hz, 1H), 3.86 (s, 3H), 3.71 (d, J=2.2 Hz, 1H), 3.68–3.64 (m, 0H), 3.60–3.51 (m, 1H), 1.96 (d, J=13.0 Hz, 1H), 1.74 (dd, J=15.3, 5.3 Hz ,4H),1.67–1.60(m,4H),1.54(dd,J＝20.4,7.7Hz,3H),1.49–1.42(m,2H),1.41–1.30(m,5H),1.27(d,J＝12.7Hz,3H),1 .13–1.05(m,4H),1.03(s,1H),1.00(d,J＝6.0Hz,3H),0.98(s,3H),0.92–0.87(m,3H),0.66–0.62(m,3H).LC-MS: [M+HH ₂ O] ⁺ =477.50.

Embodiment 138:

Compound 138: Preparation of (1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4,4-difluoro-7-hydroxy-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-one

Step 1: Dissolve compound (1R, 3aS, 3bS, 6R, 7S, 9aR, 9bS, 11aR)-7-(benzyloxy)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4,4-difluoro-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-ol 94-7 (110 mg, 0.189 mmol, 1 eq) in dichloromethane (5 mL), slowly add Dess-Martin periodinane (160 mg, 0.378 mmol, 2 eq), stir at room temperature for 3 hours, and monitor by TLC (petroleum ether: ethyl acetate = 4:1). After the reaction was completed, water was added to quench the reaction, and then the reaction mixture was extracted with ethyl acetate (50 mL). The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and dried to obtain a crude product. The crude product was separated and purified by flash chromatography (petroleum ether: ethyl acetate = 100:0 to 3:1) to obtain (1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-7-(benzyloxy)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4,4-difluoro-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-one as a white solid 138-1 (109 mg, purity: 97%, yield: 73%). ¹ H NMR (400 MHz, CDCl ₃ )δ8.12–8.05(m,2H),7.58(t,J＝7.4Hz,1H),7.45(t,J＝7.7Hz,2H),5.44(dd,J＝ 11.9,7.5Hz,1H),2.63(d,J＝13.8Hz,1H),2.42(s,1H),2.14–1.99(m,6H),1.96– 1.82(m,4H),1.79–1.60(m,7H),1.49(dd,J＝11.0,6.9Hz,3H),1.36(d,J＝5.9Hz, 5H),1.31(s,6H),1.29(s,2H),0.95(d,J＝6.5Hz,3H),0.85(s,3H),0.69(s,3H).

Step 2: At room temperature, (1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-7-(benzyloxy)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4,4-difluoro-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-one 138-1 (15 mg, 0.025 mmol, 1.0 eq) was dissolved in tetrahydrofuran (0.5 mL) and methanol (0.5 mL). Lithium hydroxide monohydrate (11 mg, 0.25 mmol, 10.0 eq) and water (0.2 mL) were added at room temperature, followed by stirring at room temperature for 30 min. TLC (petroleum ether: ethyl acetate = After the reaction was completed, the mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL × 2), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated. The crude product was separated and purified by flash chromatography (petroleum ether:ethyl acetate = 100:0 to 3:1) to obtain (1R, 3aS, 3bS, 7S, 9aR, 9bS, 11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4,4-difluoro-7-hydroxy-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-6-one 138 (6.2 mg, purity 100%, yield 52%) as a white solid.

Compound 138: ¹ H NMR (400 MHz, CDCl ₃ ) δ 4.15 (t, J = 8.2 Hz, 1H), 3.49 (d, J = 3.4 Hz, 1H), 2.55–2.42 (m, 2H), 2.11 (d, J = 8.9 Hz, 1H), 2.02 (d, J = 12.5 Hz, 2H), 1.91 (d, J = 13.0 Hz, 2H), 1.84 (d, J = 5.5 Hz, 1H), 1.73 (s, 2H), 1.67 (s, 1H), 1.62 (d, J = 11.8 Hz, 2H),1.55(d,J＝9.3Hz,2H),1.48(d,J＝7.5Hz,2H),1.37(s,1H),1.33(s,1H),1.31(s,6H),1.29(s,2H), 1.15(t,J＝9.3Hz,2H),0.94(d,J＝6.5Hz,3H),0.88(t,J＝6.7Hz,1H),0.77(s,3H),0.69(d,J＝7.5Hz,3H). ¹⁹ F NMR (377MHz, CDCl ₃ ) δ -90.22, -90.85, -111.49, -112.12, -115.16, -115.81, -116.09, -116.74. LC-MS: [MH] ⁺ = 489.35.

Embodiment 139:

Compound 139: (1R,3aR,7S,9aR,9bR,11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4-fluoro-9a,11a-dimethyl- Preparation of 2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-Tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol

Step 1: At room temperature, (3S, 4R, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-6-acetoxy-5,5-difluoro-6-methylheptane-2-yl)-10,13-dimethyl-7-oxohexadecahydro-1H-cyclopentadienyl[a]phenanthrene-3,4-diyl diacetate 94-4 (53 mg, 0.089 mmol) was dissolved in diethylaminosulfur trifluoride (2 mL), stirred at 70 ° C for 2 hours, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 4:1). After the reaction was completed, the reaction solution was diluted with ethyl acetate and slowly added dropwise to ice water (20 mL) to quench, and ethyl acetate (15 mL × 2) was extracted. The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The organic phase was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1) to give white solid acetate-(6R)-6-[(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-6,7-diacetoxy-4-fluoro-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-3,3-difluoro-2-methylhept-2-yl ester 139-1 (4.5 mg, purity: 90%, yield: 8.2%).

Step 2: At room temperature, a white solid (6R)-6-[(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-6,7-diacetoxy-4-fluoro-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-3,3-difluoro-2-methylhept-2-yl acetate 139-1 (45 mg, 0.073 mmol, 1.0 eq) was obtained. The mixture was dissolved in tetrahydrofuran (1 mL) and methanol (1 mL), and lithium hydroxide monohydrate (15 mg, 0.365 mmol, 5.0 eq) and water (0.5 mL) were added at room temperature, followed by 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 mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL × 2), and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1). A white solid (1R, 3aR, 5aR, 6R, 7S, 9aR, 9bR, 11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4-fluoro-9a, 11a-dimethyl-2,3,3a, 5,5a, 6,7,8,9,9a,9b, 10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol 139-2 (28 mg, purity: 90%, yield: 77.78%) was obtained.

Step 3: (1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4-fluoro-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol 139-2 (28 mg, 0.057 mmol, 1.0 eq) was dissolved in dichloromethane (2 mL) and tetrahydrofuran (1 mL) at room temperature, and triethylamine (35 mg, 0.057 mmol, 1.0 eq) was added. , 0.34mmol, 6.0eq), 4-dimethylaminopyridine (7mg, 0.057mmol, 1.0eq) and benzoyl chloride (24mg, 0.17mmol, 3.0eq), then stirred at room temperature for 8 hours, TLC (petroleum ether: ethyl acetate = 1: 1) monitored the reaction, after the reaction was completed, quenched with water (10mL), extracted with ethyl acetate (10mL×2), the organic phase was washed with saturated brine (20mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) and SFC (Instrument: Waters Acquity UPCC, Column: Daicel CHIRALPAK IC_3, 3.0*150mm, 3um, Mobile Phase: A/B: CO2/MeOH (0.1% DEA) = 50/50, Flow rate: 1.5ml/min, Column Temp: 37 degrees) to give a white solid (benzoic acid-(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4-fluoro-6-hydroxy-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 139-3 (15 mg, purity: 95%, yield: 44.12%, retention time t _R =1.624 min). ¹ H NMR (400 MHz, CDCl ₃ )δ8.04(d,J＝8.1Hz,2H),7.58(t,J＝7.2Hz,1H),7.45(t,J＝7.7Hz,2H),4.99(dd, J＝8.3,3.6Hz,1H),3.98(s,1H),2.25(dd,J＝34.8,4.1Hz,1H),2.08–1.95(m,3H), 1.89–1.71(m,9H),1.59–1.44(m,6H),1.31(s,6H),1.26(s,1H),1.17(d,J＝13.4 Hz,2H),1.14(s,3H),1.02(t,J＝10.2Hz,1H),0.95(d,J＝6.5Hz,3H),0.69(s,3H).

Step 4: At room temperature, (1R, 3aR, 7S, 9aR, 9bR, 11aR)-7-(benzyloxy)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4-fluoro-9a, 11a-dimethyl-2,3,3a, 5,5a, 6,7,8,9,9a, 9b, 10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6-ol 139-3 (15 mg, 0.025 mmol, 1.0 eq) was dissolved in tetrahydrofuran (0.5 mL) and methanol (0.5 mL). Lithium hydroxide monohydrate (11 mg, 0.25 mmol, 10.0 eq) and water (0.2 mL) were added at room temperature, followed by stirring at room temperature for 30 min. TLC (petroleum ether: ethyl acetate) was used to analyze the product. After the reaction was completed, the mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL×2), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The crude product was separated and purified by flash chromatography (petroleum ether:ethyl acetate=100:0 to 3:1) to give (1R,3aR,7S,9aR,9bR,11aR)-1-[(2R)-5,5-difluoro-6-hydroxy-6-methylhept-2-yl]-4-fluoro-9a,11a-dimethyl-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-6,7-diol as a white solid 139 (10 mg, purity 100%, yield 79%).

Compound 139: ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.85 (s, 1H), 3.60 (d, J = 11.0 Hz, 1H), 2.66 (t, J = 12.4 Hz, 1H), 2.14 (s, 1H), 2.01 (t, J = 11.5 Hz, 3H), 1.89 (s, 2H), 1.85 (d, J = 4.1 Hz, 1H), 1.77 (s, 2H), 1.71 (s, 1H), 1.63–1.5 5(m,4H),1.47–1.44(m,2H),1.37(s,1H),1.33(s,1H),1.30(s,6H),1.28(s,2H),1.13 –1.08(m,2H),1.05(s,3H),0.95(d,J＝6.5Hz,3H),0.88(t,J＝6.6Hz,1H),0.65(s,3H). ¹⁹ F NMR (377MHz, CDCl ₃ ) δ -109.66, -115.15, -115.80, -116.12, -116.77. LC-MS: [MH] ⁺ = 471.35.

Embodiment 143:

Compound 143: Preparation of 7-amino-24-[hydroxy(2-methoxyphenyl)methyl]-5α-cholane-3β,4β-diol

The first step: Compound (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-silanadecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-11-one 1 06-4 (50 mg, 0.075 mmol, 1.0 eq) was dissolved in a mixed solvent of tetrahydrofuran (2 mL) and methanol (1 mL), and a catalytic amount of acetic acid (0.001 mL, 0.014 mmol) and ammonium acetate (57.81 mg, 0.750 mmol, 10 eq) were added. The resulting mixture was stirred in the reaction solution at N2, 25 °C for 1 h, and then sodium cyanoborohydride [reducing agent] (47.13 mg, 0.750 mmol, 10 eq) was added. Stirring continued for 2 hours, and the reaction progress was monitored by TLC plate (dichloromethane: methanol = 10: 1). Water was added to quench, and the aqueous layer was extracted with dichloromethane (3×10 mL). The dichloromethane layers were combined and washed with saturated brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated to give a crude product, which was purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to give a white solid (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-silicon [7-nitro-1,2-d]-1,2-d]-1,2-d]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-11-amine 143-1 (30 mg, 0.043 mmol, purity: 90%, yield: 56.87%). LC-MS: [M+H] ⁺ = 668.4.

Step 2: Compound (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-silanadecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,1 2a,12b-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-11-amine 143-1 (30 mg, 0.05 mmol, 1.0 eq) was dissolved in tetrahydrofuran (5 mL), diluted hydrochloric acid (0.05 mL) was added, and the mixture was stirred at 25°C for 1 hour, and continued to stir for 2 hours. The reaction progress was monitored by TLC (dichloromethane: methanol = 5:1). Water was added to quench the mixture, and the aqueous layer was extracted with dichloromethane (3×10 mL). The dichloromethane layers were combined and washed with saturated brine (5 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography (dichloromethane: methanol = 10:1) to obtain a white Solid 7-amino-24-[hydroxy(2-methoxyphenyl)methyl]-5α-cholane-3β,4β-diol 143 (17.79 mg, 0.034 mmol, purity: 97.15%, yield: 74.92%).

Compound 143: ¹ H NMR (400 MHz, DMSO) δ7.55 (s, 1H), 7.38 (m, 1H), 7.18 (t, J = 7.8 Hz, 1H), 6.92 (m, 2H), 4.86 (s, 2H), 4.42 (d, J = 5.8 Hz, 1H), 4.17 (t, J = 6.3 Hz, 1H), 3.75 (d, J = 3.0 Hz, 3H), 3.44 (d, J = 22.4 Hz, 1H), 3. 26(m,1H),1.94(dd,J＝27.6,11.7Hz,2H),1.75(m,3H),1.59(t,J＝12.9Hz,4H),1.43(s,5H),1. 25(m,7H),1.00(s,6H),0.96(s,4H),0.86(d,J＝5.8Hz,3H),0.61(d,J＝2.5Hz,3H).LC-MS:[M+H] ⁺ =427.3.

Embodiment 144:

Compound 144: Preparation of 4-methyl-5α-cholest-3β,25-diol

Step 1: At room temperature, (3S, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-17-((R)-6-hydroxy-6-methylheptane-2-yl)-10,13-dimethyl-4-methylenehexahydro-1H-cyclopenta[a]phenanthrene-3-ol 123 (25 mg, 0.060 mmol, 1 eq) was dissolved in dichloromethane (2 mL), DMAP (2 mg, 0.016 mmol, 0.3 eq) and triethylamine (36 mg, 0.36 mmol, 6 eq) were added, cooled to 0 °C in an ice bath, and a solution of acetic anhydride (12 mg, 0.12 mmol, 2 eq) in dichloromethane (0.5 mL) was slowly added dropwise. After 10 min, the mixture was warmed to room temperature and stirred for 6 hours. The reaction solution was quenched with water and extracted with dichloromethane (10 mL x 2). The organic phases were combined and washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The product was separated by chromatography (4 g, 0-10% EtOAc/PE, 20 mL/min) to obtain (R)-6-((3S, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-3-acetoxy-10,13-dimethyl-4-methylenehexahydro-1H-cyclopentylphenanthrene-17-yl)-2-methylhept-2-yl acetate 144-1 (10 mg, purity: 90%, yield 43.3%). ¹ H NMR (400MHz, CDCl3) δ5.11(d,J＝6.7Hz,1H),4.86(s,1H),4.59(s,1H),2.12(s,3H),2.05–1.89(m,3H),1.77(dd,J＝27.1,12.6Hz,5H),1.56(d,J＝ 12.7Hz,3H),1.47–1.31(m,10H),1.21(s,6H),1.09(dd,J＝24.8,14.8Hz, 5H),0.92(d,J＝6.3Hz,3H),0.88–0.76(m,2H),0.71(s,3H),0.65(s,3H).

Step 2: (R)-6-((3S, 5R, 8S, 9S, 10R, 13R, 14S, 17R)-3-acetoxy-10,13-dimethyl-4-methylenehexahydro-1H-cyclopentylphenanthrene-17-yl)-2-methylhept-2-yl acetate 144-1 (10 mg, 0.020 mmol, 1 eq) was dissolved in room temperature. Ethyl acetate (3 mL), 10% Pd/C (20 mg) and acetic acid (0.1 mL) were added, and the mixture was stirred in a hydrogen atmosphere at room temperature for 16 hours. The reaction was detected by HNMR. After the reaction was completed, the reaction solution was diluted with ethyl acetate (10 mL), filtered through diatomaceous earth, and the filtrate was washed with saturated sodium bicarbonate solution (10 mL) and saturated brine (10 mL) in turn, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a white solid (6R)-6-((3S, 5S, 8S, 9S, 10R, 13R, 14S, 17R)-3-acetoxy-4,10,13-trimethylhexahydro-1H-cyclopentadienylphenanthren-17-yl)-2-methylheptane-2-yl acetate 144-2 (5 mg, purity: 90%, yield: 44.8%). ¹ H NMR (400MHz, CDCl ₃ )δ4.89–4.27(m,1H),2.10(s,1H),2.04(d,J＝4.5Hz,1H),1.97(s,1H),1.71(d,J＝10.9Hz,3H),1.42(d,J =3.9Hz, 6H), 1.04 (dd, J = 15.6, 9.8Hz, 7H), 0.85 (ddd, J = 20.0, 11.2, 5.5Hz, 11H), 0.64 (d, J = 4.9Hz, 3H).

Step 3: At room temperature, (6R)-6-((3S, 5S, 8S, 9S, 10R, 13R, 14S, 17R)-3-acetoxy-4,10,13-trimethylhexahydro-1H-cyclopentadienylphenanthren-17-yl)-2-methylheptane-2-yl acetate 144-2 (5 mg, 0.010 mmol, 1.0 eq) was dissolved in tetrahydrofuran (0.5 mL) and methanol (0.5 mL), and lithium hydroxide monohydrate (8 mg, 0.19 mmol, 19.2 eq) and water (0.2 mL) were added at room temperature, followed by stirring at room temperature for 2 hours and in an oil bath at 60°C for 3 hours. The reaction was detected to be complete by TLC plate (petroleum ether: ethyl acetate = 4:1, phosphomolybdic acid baking plate). After the reaction, the mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL×2), the organic phase was washed with saturated brine (20 mL), and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to give 4-methyl-5α-cholest-3β,25-diol 144 as a white solid (3.07 mg, purity: 99.6%, yield: 38.3%).

Compound 144: ¹ H NMR (400MHz, CDCl ₃ )δ3.79–2.95(m,1H),1.95(d,J＝12.7Hz,1H),1.82–1.66(m,3H),1.58(d,J＝8.3Hz,4H),1.48–1.32(m ,10H),1.27(d,J＝12.2Hz,3H),1 .21(s,6H),1.12–0.98(m,5H),0.96–0.87(m,7H),0.83(d,J＝6.3Hz,3H),0.80–0.67(m,1H),0.64(d ,J＝4.9Hz,3H),0.62–0.55(m,1H).

Embodiment 158&159:

Preparation of compound 158 [(1R, 3aS, 3bS, 4Z, 6R, 7S, 9aR, 9bS, 11aR)-6,7-dihydroxy-1-[(2R)-6-hydroxy-6-(2-methoxyphenyl)hexan-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-4-ylidene]acetonitrile and compound 159 [(1R, 3aS, 3bS, 6R, 7S, 9aR, 9bS, 11aR)-6,7-dihydroxy-1-[(2R)-6-hydroxy-6-(2-methoxyphenyl)hexan-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-4-yl]acetonitrile.

Step 1: In an ice-water bath, n-butyl lithium (0.180 mL, 0.450 mmol) solution was added dropwise to a solution of diethyl cyanomethylphosphonate (87.63 mg, 0.495 mmol) in tetrahydrofuran (5 mL). After stirring for 30 minutes, (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12aR, 12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa- 3-Silanedec-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-11-one 106-4 (30 mg, 0.045 mmol) was dissolved in tetrahydrofuran and added dropwise to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The reaction solution was quenched with saturated ammonium chloride solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and spin-dried to obtain a crude product. The crude product was purified by flash chromatography (petroleum ether/ethyl acetate = 8%) to give [(3aS,5aR,5bS,7aR,8R,10aS,10bS,11Z,12aR,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-silanadecan-9-yl]-2,2,5a,7a-tetramethyl -4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-11-ylidene]acetonitrile Colorless transparent solid 159-1 (45 mg, 0.055 mmol, 61.62%). ¹ H NMR (400 MHz, CDCl ₃ )δ7.45(d,J＝7.6Hz,1H),7.18(s,1H),6.95(d,J＝7.4Hz,1H),6.81(d,J＝8.2Hz,1H),5.14–4.99(m,2H),4.12(d,J＝7.1Hz,1H),4.07– 3.97(m,1H),3.81(s,3H),2.83(dd,J＝12.8,2.4Hz,1H),2.44(s,1H),2.28(s,1H),2.02(d,J＝13.0Hz,1H),1.93–1.77(m,4H),1.73– 1.59(m,3H),1.52(s,4H),1.42(ddd,J＝23.0,18.0,9.9Hz,7H),1.32(s,3H),1.26(dd,J＝6.3,3.5Hz,3H),1.21(s,3H),1.09(d,J＝8. 8Hz,2H),1.06–0.96(m,3H),0.89(s,9H),0.82(dd,J＝12.0,7.0Hz,2H),0.67(d,J＝6.0Hz,3H),0.02(t,J＝3.0Hz,3H),-0.14(s,3H).

Step 2: [(3aS,5aR,5bS,7aR,8R,10aS,10bS,11Z,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-silanadecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b, 11,12,12a,12b-Hexadecahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-11-ylidene]acetonitrile 159-1 (40 mg, 0.058 mmol) was dissolved in methanol (5 mL), Pd/C (20 mg, 0.094 mmol) was added, replaced with hydrogen, and the mixture was stirred at 40°C for 2 hours. TLC (petroleum ether/ethyl acetate = 5/1) was used to track the reaction solution and found that the raw material had reacted completely and there was a new point with increased polarity. Pd/C was filtered through diatomaceous earth and dried by spin drying. The crude product was purified by flash chromatography (petroleum ether/ethyl acetate = 10%) to give [(3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10bS, 12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-11-yl]acetonitrile 159-2 as a white solid (20 mg, 0.025 mmol, ^42.35 %). NMR (400MHz, CDCl3) δ7.60(d,J＝7.6Hz,1H),7.32(s,1H),7.09(d,J＝7.4Hz,1H),6.95(d,J＝8.1Hz,1H),5.26–5.16 (m,1H),4.20–4.07(m,2H),3.95(s,3H),2.69(s,2H),2.14(d,J＝12.6Hz,1H),2.02–1.78(m,7H),1.70(d,J＝15.0H z,6H),1.65(s,3H),1.57(s,1H),1.55–1.46(m,5H),1.45(s,3H),1.41(d,J＝12.9Hz,3H),1.34–1.23(m,3H),1.18 (s,3H),1.13(dd,J＝16.0,8.1Hz,2H),1.03(s,9H),0.99(s,2H),0.83(d,J＝5.8Hz,3H),0.16(s,3H),-0.00(s,3H).

Step 3: [(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-siladecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-11-yl]acetonitrile 159-2 (35 mg, 0.051 mmol) was dissolved in tetrahydrofuran (3 mL), hydrochloric acid (2 mL, 6.000 mmol) was added, and the mixture was stirred at room temperature for 2 hours. Water was added to the reaction solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and spin-dried. The crude product was purified by flash chromatography (petroleum ether/ethyl acetate = 50%) to obtain [(1R, 3aS, 3bS, 6R, 7S, 9aR, 9bS, 11aR)-6,7-dihydroxy-1-[(2R)-6-hydroxy-6-(2-methoxyphenyl)hexan-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-4-yl]acetonitrile 159 (5 mg, 0.008 mmol, 15.63%) as a white solid.

Compound 159: ¹ H NMR (400 MHz, CDCl3) δ7.31–7.27 (m, 1H), 7.26–7.21 (m, 1H), 6.96 (t, J = 7.4 Hz, 1H), 6.88 (d, J = 8.2 Hz, 1H), 4.89–4.80 (m, 1H), 3.86 (s, 3H), 3.74 (s, 1H), 3.56 (d, J = 10.7 Hz, 1H), 2.54 (d, J = 4.5 Hz, 2H), 1.99 (d, J = 12.1 Hz, 2H), 1.86 (t, J = 10.0 Hz, 2H), 1.76 ( d,J＝10.3Hz,3H),1.69–1.63(m,3H),1.51(dd,J＝12.3,7.2Hz,3H),1.43–1.32(m,6H),1.27(d,J＝12.7Hz,3H),1.10(dd,J＝17. 6,8.8Hz,4H),1.03(s,3H),0.97(dd,J＝14.0,3.6Hz,1H),0.90(dd,J＝6.4,3.6Hz,3H),0.69(d,J＝2.6Hz,3H).LCMS(ESI)[M+HH ₂ O] ⁺ =520.45,t _R =6.941min.

Step 4: [(3aS,5aR,5bS,7aR,8R,10aS,10bS,11Z,12bR)-8-[(9R)-5-(2-methoxyphenyl)-2,2,3,3-tetramethyl-4-oxa-3-silanadecan-9-yl]-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a, 10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-11-ylidene]acetonitrile 159-1 (40 mg, 0.058 mmol) was dissolved in tetrahydrofuran (3 mL), hydrochloric acid (2 mL, 6.000 mmol) was added, and the mixture was stirred at room temperature for 2 hours. Water was added to the reaction solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by flash chromatography (petroleum ether/ethyl acetate = 50%) to obtain [(1R,3aS,3bS,4Z,6R,7S,9aR,9bS,11aR)-6,7-dihydroxy-1-[(2R)-6-hydroxy-6-(2-methoxyphenyl)hexan-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-a]phenanthrene-4-ylidene]acetonitrile 158 (15 mg, 0.024 mmol, 42.03%) was obtained as a white solid.

Compound 158: ¹ H NMR (400 MHz, CDCl3) δ7.33–7.27 (m, 1H), 7.26–7.20 (m, 1H), 6.96 (t, J = 7.5 Hz, 1H), 6.89 (d, J = 8.2 Hz, 1H), 5.06 (s, 1H), 4.85 (dt, J = 9.6, 5.8 Hz, 1H), 3.86 (s, 4H), 3.62–3.52 (m, 1H), 2.73 (dd, J = 12.7, 2.3 Hz, 1H), 2.55 (t, J = 13.0 Hz, 1H), 2.24 (t, J = 10 .9Hz,1H),2.01(d,J＝12.6Hz,2H),1.85(d,J＝6.6Hz,2H),1.78–1.67(m,4H),1.43(dd,J＝15.2,4.9Hz,5H),1.32–1.22(m,4H ),1.19(s,3H),1.15–1.05(m,3H),1.04–0.95(m,2H),0.92(dd,J＝6.4,4.0Hz,3H),0.84–0.78(m,1H),0.67(d,J＝2.4Hz,3H). ¹³ C NMR (101MHz, CDCl ₃ )δ156.57,132.56,128.27,127.01,126.88,120.76,117.66,110.57,90.13,7 4.06,71.84,71.35,70.95,56.44,55.28,54.96,51.72,49.47,43.80,42.94, 38.33,37.74,37.55,36.48,36.28,35.99,35.79,35.61,35.47,35.40,31.54 ,28.06,25.64,25.07,22.70,22.56,18.74,12.13,-0.01.LCMS(ESI)[M+H-2H ₂ O] ⁺ =500.37, t _R =5.860min.

Embodiment 160:

Compound 160 Preparation of 25-hydroxy-4β-hydroxy-3β-hydroxy-5α-cholestane-7-carbonitrile.

Step 1: At room temperature, (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-tetramethyl-11-oxy-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1 ',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester I-12 (200 mg, 0.421 mmol, 1 eq) was dissolved in methanol (5 mL), cooled to 0 °C under nitrogen protection, sodium borohydride (24 mg, 0.632 mmol, 1.5 eq) was added, and then stirred at room temperature for 2 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 2:1). After the reaction was completed, it was quenched with water (30 mL), extracted with ethyl acetate (30 mL × 2), and the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and the organic phase was spin-dried to obtain a white solid (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11-hydroxy-2,2,5 a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester 160-1 (200 mg, purity: 95%, yield: 99%) was directly added to the rear.

Step 2: At room temperature, (5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11-hydroxy-2,2,5a,7a-tetramethyl-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-hexahydro-3aH-cyclopenta[1',2':1,2]phenanthro[7,8-d][1,3]dioxolan-8-yl]hexanoic acid methyl ester 160-1 (200 mg, 0.42 mmol) was dissolved in anhydrous tetrahydrofuran (5 mL), cooled to 0°C under nitrogen protection, and methylmagnesium bromide (3 M in ) was added dropwise. 2-Me-THF, 0.86 mL, 4.19 mmol, 10 eq), then stirred at room temperature for 2 hours, monitored by TLC (petroleum ether: ethyl acetate = 2: 1), after the reaction, the system was cooled to 0 ° C, quenched with saturated aqueous ammonium chloride solution (30 mL), extracted with ethyl acetate (30 mL × 2), the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1) to obtain a white solid (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-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxol-11-ol 160-2 (139 mg, purity: 95%, yield: 67%). ¹ H NMR (400MHz, CDCl ₃ )δ4.05–3.92(m,3H),1.98–1.89(m,2H),1.80(dd,J＝14.1,7.2Hz,3H),1.75–1.54(m,6H),1.48–1.33 (m,13H),1.30(s,3H),1.27(dd,J＝9.8,2.6Hz,4H),1.21(s,6H),1.19–1.10(m,4H),1.05(d,J＝7.6Hz, 3H), 0.92 (d, J = 6.5Hz, 3H), 0.68 (d, J = 8.2Hz, 3H).

Step 3: (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-hexahydro-3aH-cyclopenta[ 1',2':7,8]phenanthro[1,2-d][1,3]dioxol-11-ol 160-2 (130 mg, 0.273 mmol, 1.0 eq) was dissolved in tetrahydrofuran (10 mL), and Dess-Martin periodinane (424 mg, 0.545 mmol, 2 eq) was added at room temperature, followed by stirring at room temperature for 1 hour. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 2:1). After the reaction was completed, the mixture was quenched with ice water (50 mL), extracted with ethyl acetate (30 mL×2), and the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain a white solid (3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(2R)-6-hydroxy]-pyrrolidone (160-2). -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-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-11-one 160-3 (110 mg, purity: 95%, yield: 85%). ¹ H NMR (400MHz, CDCl ₃ )δ4.06–3.95(m,2H),2.78(t,J＝13.5Hz,1H),2.43(t,J＝11.3Hz,1H),2.27 –2.17(m,2H),1.99(dd,J＝9.6,3.3Hz,1H),1.92–1.83(m,2H),1.78–1.61( m,4H),1.53(s,3H),1.50–1.35(m,8H),1.30(s,6H),1.26(dd,J＝6.8,4.5H z,2H),1.21(s,6H),1.13–0.99(m,5H),0.92(d,J＝6.4Hz,3H),0.66(s,3H).

Step 4: Potassium tert-butoxide (106 mg, 1.95 mmol, 1.0 eq) was dissolved in dimethylformamide (3 mL) at room temperature, p-toluenesulfonylmethyl isocyanide (79 mg, 0.38 mmol, 1.5 eq) was added at room temperature, and then methanol (8.1 mg, 0.25 mmol, 1 eq) was added after stirring at room temperature for 5 minutes, and (3aS, 5aR, 5bS, 7aR, 8R, 10aS, 10b S,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-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolan-11-one 160-3 (120 mg, 0.253 mmol) , 1.0eq), stirred at room temperature for 12 hours, monitored by TLC (petroleum ether: ethyl acetate = 3: 1), quenched with water (30 mL) after the reaction, extracted with ethyl acetate (20 mL × 2), washed with saturated brine (30 mL) and dried over anhydrous sodium sulfate, concentrated the organic phase, purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15: 1), and obtained a white solid (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-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1,2-d][1,3]dioxolane-11-carbonitrile 160-4 (30 mg, purity: 90%, yield: 25%). Compound 160-4: ¹ H NMR (400 MHz, CDCl ₃ ) δ 4.05 (dt, J = 8.4, 5.1 Hz, 2H), 2.93 (dd, J = 6.6, 4.4 Hz, 1H), 2.00–1.95 (m, 2H), 1.82–1.60 (m, 10H), 1.50 (s, 3H), 1.38 (ddd, J = 15.5, 11.4, 5.4 Hz, 10H), 1.30 (s, 3H), 1.25 (s, 5H), 1.22 (s, 6H), 1.05 (s, 3H), 0.93 (t, J = 6.0 Hz, 3H), 0.75–0.62 (m, 3H).

Step 5: At room temperature, (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-hexahydro-3aH-cyclopenta[1',2':7,8]phenanthro[1, 2-d][1,3]dioxolane-11-carbonitrile 160-4 (30 mg, 0.06 mmol, 1 eq) was dissolved in tetrahydrofuran (3 mL) and 3 M hydrochloric acid (1 ml), and then stirred at room temperature for 30 min. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1:1). After the reaction was completed, it was diluted with water (10 mL), extracted with ethyl acetate (10 mL × 2), and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated. The organic phase was purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1) to obtain 25-hydroxy-4β-hydroxy-3β-hydroxy-5α-cholest-7-carbonitrile 160-5 as a white solid (15 mg, purity: 80%, yield: 50%). ¹ H NMR (400MHz, CDCl ₃ )δ3.76(s,1H),3.69–3.61(m,1H),2.93(s,1H),2.27–2.18(m,1H),2.12–1.96(m,4H),1.89(d,J＝9.6Hz,2H),1.73(dd,J＝21.6,11.8Hz,6H),1. 61(dd,J＝11.3,6.6Hz,6H),1.46(s,2H),1.37(s,3H),1.28(s,2H),1.2 5(s,3H),1.22(s,3H),1.04(s,3H),0.92(d,J＝6.5Hz,3H),0.66(s,3H).

Step 6: At room temperature, 25-hydroxy-4β-hydroxy-3β-hydroxy-5α-cholest-7-carbonitrile 160-6 (10 mg, 0.022 mmol, 1.0 eq) was dissolved in dichloromethane (2 mL), and triethylamine (7 mg, 0.07 mmol, 3.0 eq), 4-dimethylaminopyridine (2.2 mg, 0.018 mmol, 0.8 eq) and benzoyl chloride (5 mg, 0.034 mmol, 1.5 eq) were added at room temperature, followed by stirring at room temperature for 1 hour. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1:1). After the reaction was completed, it was quenched with water (10 mL), extracted with ethyl acetate (10 mL×2), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) and chiral separation (instrument: Waters Acquity UPCC, chromatographic column: Daicel CHIRALPAK IB 4.6*250mm, 5um, mobile phase: CO2/MeOH (0.1% DEA) = 60/40, flow rate: 1.5ml/min, column temperature: 37degrees) was separated to obtain white solid benzoic acid-(1R, 3aS, 3bS, 5aR, 6R, 7S, 9aR, 9bS, 11aR)-4-cyano-6-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a, 11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 160-6 (8mg, purity: 95%, yield: 80%, retention time: 1.595min). ¹ H NMR (400MHz, CDCl ₃ )δ8.04(d,J＝7.3Hz,2H),7.57(d,J＝7.4Hz,1H),7.45(t,J＝7.7Hz,2H),5.11–4.94(m ,1H),4.02(s,1H),2.94(s,1H),2.24–2.09(m,2H),1.99(d,J＝12.5Hz,2H),1.87–1. 79(m,3H),1.70(d,J＝14.1Hz,3H),1.38(dd,J＝18.1,12.4Hz,7H),1.26(d,J＝2.3Hz, 6H),1.22(s,6H),1.10(d,J＝9.7Hz,4H),0.91(dd,J＝16.7,6.4Hz,4H),0.67(s,3H).

Step 7: At room temperature, white solid benzoic acid-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4-cyano-6-hydroxy-1-[(2R)-6-hydroxy-6-methylhept-2-yl]-9a,11a-dimethylhexahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester 160-6 (20 mg, 0.045 mmol, 1 eq) was dissolved in tetrahydrofuran (0.5 mL) and methanol (0.5 mL), and lithium hydroxide monohydrate (11 mg, 0.25 mmol, 1 eq) was added at room temperature. 10.0eq) and water (0.2mL), then stirred at room temperature for 30min, the reaction was monitored by TLC (petroleum ether:ethyl acetate=10:1). After the reaction was completed, it was diluted with water (10mL), extracted with ethyl acetate (10mL×2), the organic phase was washed with saturated brine (20mL), and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1) to give 25-hydroxy-4β-hydroxy-3β-hydroxy-5α-cholestan-7-carbonitrile 160 (12mg, purity: 98.73%, yield: 60%) as a white solid.

Compound 160: ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.76 (s, 1H), 3.66 (s, 1H), 2.93 (dd, J = 4.5, 2.3 Hz, 1H), 2.28 (dd, J = 37.9, 7.5 Hz, 2H), 2.16–2.04 (m, 2H), 2.04–1.94 (m, 3H), 1.89 (dd, J = 8.1, 4.5 Hz, 1H), 1.73 (dd, J = 8.7, 4.9 Hz, 3H), 1. 66–1.58(m,4H),1.49–1.40(m,6H),1.33(s,4H),1.22(s,6H),1.09(dd,J＝12.7,5.1Hz,2H),1 .04(s,3H),0.92(d,J＝6.5Hz,3H),0.89(d,J＝6.7Hz,2H),0.66(s,3H).LC-MS: [MH]+＝445.36.

Effect Example 1 Inhibitory Effect of the Compounds of the Invention on SREBP Pathway

1.1: SREBP luciferase reporter gene system

Huh-7/SRE-luc cells were incubated in sterol-deficient medium (5% delipidated serum, 2μM lovastatin, 10μM mevalonic acid) and treated with corresponding concentrations of compounds for 16 hours. Among them, Huh-7/SRE-luc cells are cells that stably express LDLR promoter-luciferase and green fluorescent protein (GFP) in the human hepatoma cell line Huh-7. The cells were obtained by transfecting Huh-7 with pLDLR-promotor-luciferase and pEGFP-N1 plasmids and screening with G418 antibiotic selection pressure. Among them, Huh-7 cells were purchased from ATCC, pLDLR-promotor-luciferase plasmids were obtained by molecular cloning methods, and pEGFP-N1 plasmids were purchased from Addgene.

After the 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 such instruments). The fluorescence intensity of green fluorescent protein (EGFP) was also measured by the above-mentioned BioTek microplate reader (including but not limited to such instruments) and used as an internal reference. The ratio of SRE-driven luciferase activity divided by the fluorescence intensity of green fluorescent protein was used as an indicator of SREBP pathway activity. The test data of each test compound was analyzed by prism software (the compound inhibition rate was calculated with the DMSO treatment group as a reference, and the Prism log (inhibitor) VS. response--Variable slope (four-parameter) model was used for fitting, and the calculation formula was: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)*HillSlope))) to obtain the IC50 parameter of the compound.

1.2 Inhibitory effect of the compounds of the present invention on the SREBP pathway

The inhibitory effect of the compounds of the present invention on the SREBP pathway was tested by the method of biological test example 1.1, and the concentration gradient of each compound was designed to be 0.01, 0.03, 0.1, 0.3, 1.0, 3.0, 10 μM, and the control was the solvent DMSO. The IC ₅₀ values of some compounds are shown in Table 1.

Table 1

Effect Example 2: Mouse liver microsome metabolic stability experiment

Prepare PB solution (100mM), MgCl2 solution (300mM), NADPH solution (2.04mM) and mouse liver microsome (SHQY item number: M1000 batch number: 2110330) solution (1.00mg/mL) respectively, then prepare compound and testosterone (positive control, half-life is 3.1 minutes) stock solution with dimethyl sulfoxide (DMSO), dilute to 100μM with methanol for sample incubation, and store at -10～-30℃. Then add 100μL of mouse liver microsome solution to the 96-well sample plate. For the sample at 0min, first add acetonitrile solution containing internal standard; secondly add 2μL 100μM working solution; then add 98μL 2.04mM NADPH preheated to 37℃. For samples of 5min, 15min, 30min, 45min and 60min, first add 2μL 100μM working solution; then add 98μL 2.04mM NADPH preheated to 37℃ to start the reaction; after reaching the corresponding time point, add acetonitrile solution containing internal standard to terminate the reaction. For samples of NCF60, first add 2μL 100μM working solution; then add 98μL PB preheated to 37℃ to start the reaction; after reaching the corresponding time point, add acetonitrile solution containing internal standard to terminate the reaction. During the experiment, the samples were incubated in a 37℃ water bath. All samples were shaken on an oscillator at 800 rpm for 10min, and then centrifuged at 3200g for 20min. The supernatant was aspirated, diluted with deionized water in the corresponding ratio, and analyzed by LC-MS/MS injection. The residual rate of the compound at each time point was obtained by dividing the peak area ratio of the compound at 5, 15, 30, 45 and 60 min by the peak area ratio at 0 min, and then the half-life (T1/2) of the compound was calculated using Excel.

Table 2: Mouse liver microsome stability results of some compounds in the examples

Effect Example 3: Testing the solubility of the compounds of the present invention in fasting state simulated intestinal fluid (FaSSIF)

Dissolve the compound powder in DMSO and prepare it into a 10.0mM stock solution for use. Weigh 0.021g NaOH, 0.198g NaH2PO4 and 0.309g NaCl, add them to 50.0mL ultrapure water, mix well, adjust the pH to 6.5 with HCl or NaOH, then add 0.112g FaSSIF powder, let stand at room temperature for 2 hours after complete dissolution. Take a certain volume of compound stock solution and add it to a certain volume of FaSSIF to prepare a 300μM sample solution, mix well at room temperature and 1000rpm for 1 hour. After 1 hour, centrifuge the sample and take a certain volume of supernatant, add stop solution, precipitate the sample, mix well and centrifuge for 20 minutes. Then take the supernatant and dilute it with diluent and analyze it by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The concentration of the compound to be tested in the sample is quantitatively determined by LC-MS/MS.

Table 3: FASSIF solubility results of some compounds in the examples

Effect Example 4: Experiment on the inhibition of FASN gene expression by compounds

Huh-7 cells (purchased from ATCC) were cultured in a 37°C, 5% CO2 incubator using DMEM medium (containing 10% fetal bovine serum LONSERA S711-001S). After 24 hours, the original medium was removed and replaced with DMEM starvation medium (containing 5% lipoprotein-free serum (LPDS), 2μM lovastatin (-lovastatin), 10μM mevalonate (mevalonate)). The control group was added with dimethyl sulfoxide (DMSO), and the compound treatment group was added with different concentrations of compounds. The highest concentration of the compound was 3000nM, 3-fold dilution, a total of 5 concentrations. Culture for 16 hours. Cell lysis, cell to cDNA reverse transcription, and qPCR detection of the expression of SREBP target gene fatty acid synthase (FASN) gene (FASN gene forward primer sequence: 5'-AAGGACCTGTCTAGGTTTGATGC-3', reverse primer sequence: 5'-TGGCTTCATAGGTGACTTCCA-3') were performed. Calculate IC50. The test data of each test compound was analyzed by prism software (the compound inhibition rate was calculated with the DMSO treatment group as a reference, and the Prism log (inhibitor) VS. response--Variable slope (four-parameter) model was used for fitting, and the calculation formula was Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)*HillSlope))) to obtain the IC50 parameters of the compound.

Table 4: Inhibition results of FASN gene expression by compounds of some examples

In the present invention, the structure of comparative compound 1 is

Although the specific embodiments of the present invention are described above, it should be understood by those skilled in the art that these are only examples, and various changes or modifications may be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the protection scope of the present invention is limited by the appended claims.

Claims (10)

A compound of formula I, II or III or a pharmaceutically acceptable salt thereof:
in, R ^4a is H; R ^4b is halogen, CN, -CH ₂ -CN, -CH ₂ -OH, -CH(CH ₃ )-OH, -COOH, -CH ₂ -COOH, C ₁ -C ₆ haloalkyl, -CH ₂ -OAc or NH ₂ ; or, R ^4a and R ^4b together with the carbon atoms to which they are attached form When there is a single bond between carbon atom No. 7 and carbon atom No. 8, R ^7a and R ^7b are independently H, halogen, CN, -CH ₂ -OH, -COOH, -CH ₂ -COOH, NH ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl or -CH ₂ -CN; or, R ^7a and R ^7b together with the carbon atoms to which they are connected form R ^8a is H; When there is a double bond between carbon atoms 7 and 8, R ^7a does not exist, R ^7b is a halogen; R ^8a does not exist; R ²¹ is n1 and n3 are independently 2, 3, 4 or 5; m1 and m3 are independently 0, 1, 2, 3, 4 or 5; Ring A and Ring C are independently C ₆ -C ₁₀ aryl groups, "heteroatoms are selected from 1, 2 or 3 of N, O and S, and the number of heteroatoms is 1, 2 or 3 5-10 membered heteroaryl" or C ₃ -C ₆ cycloalkyl; ^RA and ^RC are independently halogen, _C1 - _C6 alkyl, C1- _{C6 haloalkyl} , _C1 - _C6 alkoxy or _C1 - _C6 haloalkoxy; ^R1 is or NR ^1a R ^1b ; R ^1a and R ^1b are independently H or C ₁ -C ₆ alkyl; R ^4c is H; R ^4d is H or OH; R ^7c is H, R ^7d is independently CN, -CH ₂ -OH, -COOH, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, -CH ₂ -COOH, NH ₂ , -CH ₂ -CN or Cl; or, R ^7c and R ^7d together with the carbon atom to which they are attached form R ²² is n2 is 2, 3, 4 or 5; m2 is 0, 1, 2, 3, 4 or 5; Ring B is C ₆ -C ₁₀ aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or C ₃ -C ₆ cycloalkyl; ^RB is independently _{halogen, C1-C6 alkyl, C1-C6 haloalkyl , C1 - C6} alkoxy or _C1 - _C6 haloalkoxy; R ^3a is H or C ₁ -C ₆ alkyl; R ^3b is -O-(CH ₂ )nR ^3-1 , NR ^3-2 R ^3-3 or OH; when R ^3b is OH, R ^3a is C ₁ -C ₆ alkyl; n is 1, 2, 3 or 4; R ^3-1 is OH or -C(O)OR ^3-1a ; R ^3-1a is H or C ₁ -C ₆ alkyl; R ^3-2 and R ^3-3 are independently H or -S(O) ₂ R ^3-2a ; R ^3-2a is C ₁ -C ₆ haloalkyl or C ₁ -C ₆ alkyl; R ^4e is H; R ^4f is H, OH or -O-(CH ₂ )mOR ^4-1 ; or R ^4e and R ^4f together with the carbon atoms to which they are attached form m is 1 or 2; R ^4-1 is C ₁ -C ₆ alkyl; A carbon atom with an "*" indicates that when it is a chiral carbon atom, it is independently in the R configuration, the S configuration, or a mixture of the two; A carbon atom with a "#" indicates that when it is a chiral carbon atom, it is independently in the R configuration, the S configuration, or a mixture of the two; A carbon atom with "&" indicates that when it is a chiral carbon atom, it is independently in R configuration, S configuration, or a mixture of the two; The compound shown in formula I is not the following compound:
and its isomers; The compound shown in formula II is not the following compound: and its isomers; The compound shown in formula III is not the following compound: and its isomers. The compound of formula I, II or III or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that one or more of the following conditions are met: (1) In R ^4b , the halogen is independently F, Cl, Br or I; for example, Br; (2) In R ^4b , the C ₁ -C ₆ haloalkyl group is independently CHF ₂ , CH ₂ F or CF ₃ ; (3) In R ^7a and R ^7b , the halogen is independently F, Cl, Br or I; preferably F; (4) In R ^7a and R ^7b , the C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl; (5) In R ^7a and R ^7b , the C ₁ -C ₆ haloalkyl group is independently CHF ₂ , CH ₂ F or CF ₃ , for example CHF ₂ ; (6) In Ring A and Ring C, the C ₆ -C ₁₀ aryl group is independently phenyl or naphthyl, preferably phenyl; (7) In Ring A and Ring C, the “heteroatoms selected from 1, 2 or 3 of N, O and S, 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms” is independently “heteroatoms selected from 1, 2 or 3 of N, O and S, 5-6 membered or 9-10 membered heteroaryl group having 1 or 2 heteroatoms”, for example, pyridyl (e.g. ); (8) In Ring A and Ring C, the C ₃ -C ₆ cycloalkyl group is independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; (9) In ^RA and ^RC , the halogen is independently F, Cl, Br or I; preferably F or Cl, such as F; (10) In ^RA and ^RC , the C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl; (11) In ^RA and ^RC , the C ₁ -C ₆ haloalkyl group is independently CHF ₂ , CH ₂ F or CF ₃ ; (12) In ^RA and ^RC , the C ₁ -C ₆ alkoxy group is independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, preferably methoxy or ethoxy; (13) In ^RA and ^RC , the C ₁ -C ₆ haloalkoxy group is independently -OCHF ₂ , -OCH ₂ F or -OCF ₃ ; preferably -OCF ₃ ; (14) In R ^1a and R ^1b , the C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl; (15) In formula I, the carbon atom with "*" indicates that when it is a chiral carbon atom, it is in S configuration; (16) In R ^7c and R ^7d , the C ₁ -C ₆ alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl; (17) In R ^7c and R ^7d , the C ₁ -C ₆ haloalkyl group is independently CHF ₂ , CH ₂ F or CF ₃ , for example CHF ₂ ; (18) In ring B, the C ₆ -C ₁₀ aryl group is phenyl or naphthyl, preferably phenyl; (19) In ring B, the “heteroatoms selected from 1, 2 or 3 of N, O and S, 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms” is “heteroatoms selected from 1, 2 or 3 of N, O and S, 5-6 membered or 9-10 membered heteroaryl group having 1 or 2 heteroatoms”, for example, pyridyl (e.g. ); (20) In ring B, the C ₃ -C ₆ cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; (21) In ^RB , the halogen is independently F, Cl, Br or I; preferably F or Cl, such as F; (22) In ^RB , the _C1 - _C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl; (23) In ^RB , the C ₁ -C ₆ haloalkyl group is independently CHF ₂ , CH ₂ F or CF ₃ ; (24) In ^RB , the _C1 - _C6 alkoxy group is independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, preferably methoxy or ethoxy; (25) In ^RB , the C ₁ -C ₆ haloalkoxy group is independently -OCHF ₂ , -OCH ₂ F or -OCF ₃ ; preferably -OCF ₃ ; (26) In formula II, the carbon atom with “*” indicates that when it is a chiral carbon atom, it is in S configuration; (27) In formula II, the carbon atom with “#” indicates that when it is a chiral carbon atom, it is in R configuration; (28) In R ^3a , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, methyl; (29) In R ^3-1a , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example tert-butyl; (30) In R ^3-2a , the C ₁ -C ₆ haloalkyl group is CHF ₂ , CH ₂ F or CF ₃ , for example CF ₃ ; and (31) In R ^4-1 , the C ₁ -C ₆ alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example, ethyl. The compound of formula I, II or III or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that one or more of the following conditions are met: (1) In formula (I), for (2) R ^4a is H; R ^4b is halogen (e.g., Br), CN, -CH ₂ -CN, -CH ₂ -OH, -CH(CH ₃ )-OH, -COOH, -CH ₂ -COOH, -CH ₂ -OAc, or NH ₂ ; or, R ^4a and R ^4b together with the carbon atoms to which they are attached form (3) When there is a single bond between carbon atom No. 7 and carbon atom No. 8, R ^7a and R ^7b are independently H or halogen (e.g., F), preferably, R ^7a and R ^7b are both H or halogen (e.g., F); (4) When there is a double bond between carbon atoms 7 and 8, R ^7a does not exist, R ^7b is a halogen (e.g., F); R ^8a does not exist; (5) R ²¹ is (6) Ring A and Ring C are independently C ₆ -C ₁₀ aryl or "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S"; (7) ^RA and ^RC are independently halogen, _C1 - _C6 alkoxy or _C1 - _C6 haloalkoxy; (8) n1 is 3; (9) n3 is 3; (10) m1 is 0, 1, 2 or 3; (11) m3 is 0, 1, 2 or 3; (12) In formula II, for (13) R ^7c is H, R ^7d is independently CN, -CH ₃ , -CH ₂ -OH, -COOH, -CHF ₂ , -CH ₂ -COOH, NH ₂ , -CH ₂ -CN or Cl; or, R ^7c and R ^7d together with the carbon atoms to which they are attached form (14) R ²² is (15) Ring B is a C ₆ -C ₁₀ aryl group or a "5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S"; (16) ^RB is independently halogen, _C1 - _C6 alkoxy or _C1 - _C6 haloalkoxy; (17) n2 is 3; (18) m2 is 0, 1, 2 or 3; (19) In formula III, for (20) n is 1, 2 or 3; (21) R ^3-2 is H, R ^3-3 is -S(O) ₂ R ^3-2a ; and (22)m is 1. The compound of formula I, II or III or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that one or more of the following conditions are met: (1) R ^4a is H; R ^4b is Br, CN, -CH ₂ -CN, -CH ₂ -OH, -CH(CH ₃ )-OH, -COOH, -CH ₂ -OAc or NH ₂ ; or, R ^4a and R ^4b together with the carbon atoms to which they are attached form (2) When there is a single bond between carbon atoms 7 and 8, R ^7a and R ^7b are both H or halogen (e.g., F); (3) R ^7c is H, R ^7d is independently CN, -CH ₂ -OH, -CH ₃ , NH ₂ , -CH ₂ -CN, -CF ₂ H, -COOH, or R ^7c and R ^7d together with the carbon atom to which they are attached form (4) R ^3a is H, R ^3b is -NH ₂ , Alternatively, R ^3a is methyl and R ^3b is OH; and (5) R ^4e is H; R ^4f is H, OH or Or R ^4e and R ^4f together with the carbon atoms to which they are attached form The compound of formula I, II or III or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that one or more of the following conditions are met: (1) for (2) for (3) In formula (1), for (4) for (5) In formula II, for (6) In formula III, for The compound of formula I, II or III or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that one or more of the following conditions are met: (1) R ²¹ is (2) R ²² is The compound of formula I, II or III or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that it is any of the following compounds or a pharmaceutically acceptable salt thereof:








A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient. A use of a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 8 in the preparation of a medicament for preventing and/or treating a disease, wherein the disease is obesity, hyperlipidemia, fatty liver, diabetes, atherosclerosis, cardiovascular and cerebrovascular diseases, liver cancer or skin damage. A use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 8 in the preparation of a medicament for inhibiting the SREBP pathway.