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WO2025020190A1 - 甾类化合物、其制备方法和应用 - Google Patents

甾类化合物、其制备方法和应用 Download PDF

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Publication number
WO2025020190A1
WO2025020190A1 PCT/CN2023/109658 CN2023109658W WO2025020190A1 WO 2025020190 A1 WO2025020190 A1 WO 2025020190A1 CN 2023109658 W CN2023109658 W CN 2023109658W WO 2025020190 A1 WO2025020190 A1 WO 2025020190A1
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Prior art keywords
mmol
ethyl acetate
independently
cyclopenta
reaction
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PCT/CN2023/109658
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English (en)
French (fr)
Inventor
刘金镖
唐静洁
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Cholesgen Shanghai Co Ltd
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Cholesgen Shanghai Co Ltd
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Priority to PCT/CN2023/109658 priority Critical patent/WO2025020190A1/zh
Priority to TW113127742A priority patent/TW202508555A/zh
Priority to PCT/CN2024/107810 priority patent/WO2025021195A1/zh
Publication of WO2025020190A1 publication Critical patent/WO2025020190A1/zh
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane

Definitions

  • the present invention relates to a steroid compound, a preparation method and application thereof.
  • 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.
  • SCAP SREBP cleavage-activating protein
  • S1P Site-1protease
  • S2P Site-2protease
  • SREBP protein The splicing and maturation of SREBP protein is strictly regulated by the level of intracellular sterols (such as cholesterol and 25-hydroxycholesterol).
  • intracellular sterols such as cholesterol and 25-hydroxycholesterol.
  • 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.
  • Insig Insulin-induced gene
  • the active form of SREBP in the nucleus increases, promoting cellular lipid synthesis.
  • 25-hydroxycholesterol 25-hydroxylcholesterol, 25-HC
  • 25-HC directly binds to Insig and induces the binding of SCAP and Insig.
  • 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.
  • 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.
  • free cholesterol accumulation has been reported as a key driver of the transition from simple steatosis to aggressive steatohepatitis.
  • 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.
  • SREBPs 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.
  • 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.
  • 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:
  • 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
  • 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;
  • R 7a 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;
  • n1 and n3 are independently 2, 3, 4 or 5;
  • n1 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;
  • 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").
  • the halogen is independently F, Cl, Br or I; for example, Br.
  • the C 1 -C 6 haloalkyl is independently CHF 2 , CH 2 F or CF 3 .
  • the halogen is independently F, Cl, Br or I; preferably F.
  • the C 1 -C 6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.
  • the C 1 -C 6 haloalkyl is independently CHF 2 , CH 2 F or CF 3 , for example CHF 2 .
  • the C 6 -C 10 aryl group is independently phenyl or naphthyl, preferably phenyl.
  • 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. ).
  • the C 3 -C 6 cycloalkyl group is independently cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
  • the halogen is independently F, Cl, Br or I; preferably F or Cl, such as F.
  • the C 1 -C 6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.
  • the C 1 -C 6 haloalkyl is independently CHF 2 , CH 2 F or CF 3 .
  • 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.
  • the C 1 -C 6 haloalkoxy group is independently -OCHF 2 , -OCH 2 F or -OCF 3 ; preferably -OCF 3 .
  • 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.
  • the carbon atom with "*" indicates that when it is a chiral carbon atom, it is in the S configuration.
  • 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
  • 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
  • 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);
  • R 7a 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.
  • R 21 is For example
  • 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".
  • RA and RC are independently halogen, C 1 -C 6 alkoxy, or C 1 -C 6 haloalkoxy.
  • n1 is 3.
  • n3 is 3.
  • m1 is 0, 1, 2 or 3.
  • m3 is 0, 1, 2 or 3.
  • R 21 is
  • the present invention also provides a compound represented by formula II or a pharmaceutically acceptable salt thereof:
  • 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
  • n2 is 2, 3, 4 or 5;
  • n2 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 C 1 -C 6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.
  • the C 1 -C 6 haloalkyl is independently CHF 2 , CH 2 F or CF 3 , such as CHF 2 .
  • the C 6 -C 10 aryl group is phenyl or naphthyl, preferably phenyl.
  • 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 ).
  • the C 3 -C 6 cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
  • the halogen is independently F, Cl, Br or I; preferably F or Cl, such as F.
  • the C1 - C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isopropyl, butyl, sec-butyl or tert-butyl.
  • the C 1 -C 6 haloalkyl is independently CHF 2 , CH 2 F or CF 3 .
  • 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.
  • the C 1 -C 6 haloalkoxy group is independently -OCHF 2 , -OCH 2 F or -OCF 3 ; preferably -OCF 3 .
  • the carbon atom with "*" indicates that when it is a chiral carbon atom, it is in the S configuration.
  • the carbon atom with "#" indicates that when it is a chiral carbon atom, it is in the R configuration.
  • 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
  • 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
  • R22 is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • 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".
  • RB is independently halogen, C1 - C6 alkoxy, or C1 - C6 haloalkoxy.
  • n2 is 3.
  • m2 is 0, 1, 2 or 3.
  • R22 is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • the present invention also provides a compound represented by formula III or a pharmaceutically acceptable salt thereof:
  • 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 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 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 C 1 -C 6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as methyl.
  • 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.
  • the C 1 -C 6 haloalkyl group is CHF 2 , CH 2 F or CF 3 , such as CF 3 .
  • the C 1 -C 6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, such as ethyl.
  • n 1, 2, or 3.
  • R 3-2 is H and R 3-3 is -S(O) 2 R 3-2a .
  • m is 1.
  • R 3a is H
  • R 3b is Alternatively, R 3a is methyl and R 3b is OH.
  • 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 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.
  • substitution or “substituent” means that the hydrogen atom in the group is replaced by a specified group.
  • 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.
  • variable e.g, RA
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • the cycloalkyl group is monocyclic.
  • the cycloalkyl group is polycyclic (eg, cyclic, spirocyclic, or bridged).
  • C 6-10 aryl refers to phenyl or naphthyl.
  • 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,
  • 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.
  • 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.
  • the term "subject” includes any animal, preferably a mammal, and more preferably a human.
  • 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.
  • 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.
  • diseases such as obesity, hyperlipidemia, fatty liver, diabetes, atherosclerosis, cardiovascular and cerebrovascular diseases, liver cancer, skin damage, etc.
  • Step 1 Dissolve (22E, 24S)-stigmaster-6(5), 22(23)-diene-3 ⁇ -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.
  • acetic anhydride 3.4 mL, 36.35 mmol, 3.0 eq
  • 4-dimethylaminopyridine 300 mg, 2.42 mmol, 0.2 eq
  • triethylamine 8 mL, 60.57 mmol, 5.0 eq
  • 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.
  • 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.
  • 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
  • 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.
  • 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- ⁇ 5-methylphosphine) methyl acetate (51.89 g, 155.21 mmol, 6.0 eq) and stir the reaction system at 90 ° C for 18 h.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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 , stir
  • 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.
  • 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.
  • 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.
  • 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.
  • 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-te
  • 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.
  • 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.
  • 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.
  • 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.
  • 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).
  • 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.
  • 5R -5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-hydroxy-9a,11a-dimethyl-2,3,3a,3b
  • 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.
  • 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.
  • 5R -5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-9a,11a-dimethyl-7-oxoylidene hexa
  • 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.
  • 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.
  • 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.
  • Step 4 In a 50 mL round-bottom flask at room temperature, 3 ⁇ - ⁇ [dimethyl(2-methylprop-2-yl)silyl]oxy ⁇ -25-hydroxycholest-4 ⁇ -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 refluxed for 0.5 h.
  • the reaction solution was poured into water and extracted with ethyl acetate (20 mL x 2). The organic phases were combined and concentrated.
  • the reaction solution was poured into water and extracted with ethyl acetate (20 mL x 2). The organic phases were combined and concentrated.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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
  • 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:e
  • 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.
  • 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).
  • 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.
  • 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.
  • 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.).
  • 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.
  • 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.
  • 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.
  • 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-cyclopen
  • 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),
  • 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,
  • 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.
  • 5R -5-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9a,11a-dimethylhexahydro-1H-cyclopenta[
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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).
  • 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.
  • 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.
  • 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.
  • 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.
  • 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 drop
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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
  • 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,
  • 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).
  • 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 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.
  • 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.
  • 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).
  • 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).
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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
  • 4-methylbenzenesulfonyl chloride (824 mg, 4.32 mmol, 10 eq) was added thereto. The mixture was stirred at room temperature for 5 days.
  • 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.
  • 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.
  • 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
  • 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 hydroper
  • 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.
  • 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.
  • 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.
  • 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.
  • Embodiment 109 is a diagrammatic representation of Embodiment 109.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 5R -5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-acetoxy-6-hydroxy-9
  • 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.
  • 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).
  • Step 5 In a 50 mL round-bottom flask, 4-methylidene cholesteryl-3 ⁇ ,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.
  • 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
  • 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.
  • 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).
  • 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.
  • 1,2-dichloroethane 60 mL
  • Dess-Martin periodinane 1838.04 mg, 4.335 mmol
  • 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.
  • 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.
  • 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.
  • 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.
  • Embodiment 123 is a diagrammatic representation of Embodiment 123.
  • 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).
  • 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.
  • 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.
  • 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).
  • 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.
  • Embodiment 137
  • 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
  • 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.
  • Embodiment 138
  • 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).
  • Embodiment 139
  • 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.
  • 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.
  • 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.
  • 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.
  • Embodiment 143
  • 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
  • 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.
  • Embodiment 144
  • 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.
  • 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.
  • 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%).
  • 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.
  • Embodiment 158&159
  • 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).
  • 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.
  • 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.
  • 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.
  • Embodiment 160 is a diagrammatic representation of Embodiment 160.
  • 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.
  • methanol 5 mL
  • sodium borohydride 24 mg
  • 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.
  • 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.
  • 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][
  • 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.
  • 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.
  • Huh-7/SRE-luc cells were incubated in sterol-deficient medium (5% delipidated serum, 2 ⁇ M lovastatin, 10 ⁇ M mevalonic acid) and treated with corresponding concentrations of compounds for 16 hours.
  • 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.
  • 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.
  • 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 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 ⁇ M, and the control was the solvent DMSO.
  • the IC 50 values of some compounds are shown in Table 1.
  • PB solution 100mM
  • MgCl2 solution 300mM
  • NADPH solution 2.04mM
  • mouse liver microsome SHQY item number: M1000 batch number: 2110330
  • compound and testosterone positive control, half-life is 3.1 minutes
  • DMSO dimethyl sulfoxide
  • LC-MS/MS liquid chromatography-tandem mass spectrometry
  • 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), 2 ⁇ M lovastatin (-lovastatin), 10 ⁇ M 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.

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Abstract

本发明公开了一种甾类化合物、其制备方法和应用。具体公开了具有如式（I）、（II）或（III）等所示结构的甾类化合物或其药学上可接受的盐。本发明的化合物具有SREBP通路抑制活性，可用于预防和/或治疗肥胖、高脂血症、脂肪肝、糖尿病、动脉粥样硬化症、心脑血管疾病、肝癌、皮肤损伤等疾病。

Description

甾类化合物、其制备方法和应用 技术领域
本发明涉及一种甾类化合物、其制备方法和应用。
背景技术
随着生活方式的转变，包括高热量食物和高糖饮料摄入、缺乏运动和体力活动等，在全球范围内，以高脂血症、肥胖、2型糖尿病以及脂肪肝为代表的代谢性疾病已经成为日益严重的健康问题。其中脂肪肝已成为欧美和我国富裕地区慢性肝病的重要病因，普通成人单纯性肝脏脂质堆积患病率10％～30％，其中10％～20％为脂肪性肝炎，后者10年内肝硬化、肝癌发病率达25％。然而截止目前，脂肪肝病理生理机制尚未完全阐明，临床上仍缺乏有效而特异的治疗药物。已知血液和肝脏中胆固醇、甘油三酯等脂质累积是引起高脂血症的主要原因，而高脂血症又是引起动脉粥样硬化、脑中风和脂肪性肝疾病的重要致病因素。因此，以降低脂质为导向，研发靶向脂质代谢调控通路的新型药物，正日渐成为新型代谢性疾病药物研发的重要方向。
已知哺乳动物细胞的脂质合成途径是调控脂质代谢平衡的重要因素。调控胆固醇和脂肪酸合成的关键因子是一类转录因子蛋白甾醇反应元件结合蛋白SREBP(Sterol-Regulatory Element Binding Protein)。这一类蛋白质的前体首先在内质网(ER)上合成，前体通过SREBP切割激活蛋白(SCAP，SREBP cleavage-activating protein)转运到高尔基体，然后经过两种蛋白酶(Site-1protease(S1P)和Site-2protease(S2P))酶切，释放其N端的活性结构域，进入细胞核发挥转录因子作用，与靶基因启动子区的SREBP反应元件(SRE)结合，启动下游基因的表达。SREBP蛋白的剪切成熟严格受细胞内甾醇(如胆固醇、25-羟胆固醇)水平的调控。当细胞在内质网中积累到充足的胆固醇时，胆固醇和SCAP结合，并改变SCAP的构象，引起SCAP-SREBP复合物结合蛋白Insig(Insulin-induced gene)，从而阻断SREBP向高尔基体的转运和随后SREBP的激活。反之，核内活性形式的SREBP增多，促进细胞脂质合成。除了胆固醇，25-羟胆固醇(25-hydroxylcholesterol，25-HC)是另一个强效的SREBP通路的内源性抑制剂。和胆固醇结合SCAP不同的是，25-HC直接结合Insig，并诱导SCAP和Insig结合。
前期研究发现抑制SREBP通路是预防和/或治疗肥胖、高脂血症、脂肪肝、动脉粥样硬化、糖尿病等代谢性疾病和心脑血管疾病、皮肤损伤、肝癌等疾病的有效策略和方法。
针对高脂血症，其发病机制主要是饮食或基因突变等因素引起的脂质合成增加或者脂质转运异常导致血液胆固醇和脂肪酸等脂质的过量累积。目前临床以他汀类药物和贝特类药物为主要调脂药物，其中他汀类药物的作用机理即是通过抑制细胞胆固醇合成途径同时促进血液胆固醇逆向运输实现的。表明靶向细胞脂质合成途径关键因子是有效降低脂质水平的重要手段。
目前为止，针对脂肪性肝病还没有被批准的治疗药物，因此确定治疗靶点和开发新的有效疗法是非常重要的。已知脂肪性肝病的发病机制涉及多种危险因素，如甘油三酯以脂滴形式积累可能引发的脂肪变性，细胞中异常增加的胆固醇和脂肪酸，能引起内质网压力和线粒体功能紊乱，从而导致细胞 死亡、炎症和纤维化。其中，游离胆固醇积累被报道为单纯脂肪变性向侵袭性脂肪性肝炎转变的关键驱动因素。其次，建立脂肪肝小鼠模型，简单的无胆固醇高脂饮食即使在长时间喂养后也只能诱导脂肪变性，而在饮食中添加1-2％的胆固醇是实现炎症和纤维化的必要条件因此，降低胆固醇可能成为脂肪性肝病的一种新的治疗策略。已有研究显示：在脂肪肝患者和脂肪肝小鼠模型中发现SREBP异常激活；小鼠肝脏特异性Scap的缺失或敲除，可以消除所有SREBPs的激活，从而阻止脂肪肝和高脂血症的发生。此外，最近的研究表明内质网应激诱导的SREBP异常激活促进了脂肪生成和脂肪肝。因此，这些证据表明，通过抑制SREBP通路降低肝脏的甘油三酯和胆固醇水平是一种预防和/或治疗包括脂肪肝在内的代谢紊乱的有效策略。
发明内容
本发明所要解决的技术问题是提供对SREBP通路具有抑制活性的新化合物。
本发明提供了一种式I所示化合物或其药学上可接受的盐：
其中，
R4a为H；R4b为卤素、CN、-CH2-CN、-CH2-OH、-CH(CH3)-OH、-COOH、-CH2-COOH、C1-C6卤代烷基、-CH2-OAc或NH2；或者，R4a和R4b以及它们连接的碳原子一起共同形成
当7号碳原子和8号碳原子之间为单键时，R7a和R7b独立地为H、卤素、CN、-CH2-OH、-COOH、-CH2-COOH、NH2、C1-C6烷基、C1-C6卤代烷基或-CH2-CN；或者，R7a和R7b以及它们连接的碳原子一起共同形成R8a为H；
当7号碳原子和8号碳原子之间为双键时，R7a不存在，R7b为卤素；R8a不存在；
R21为
n1和n3独立地为2、3、4或5；
m1和m3独立地为0、1、2、3、4或5；
环A和环C独立地为C6-C10芳基、“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个、2个或3个的5-10元杂芳基”或C3-C6环烷基；
RA和RC独立地为卤素、C1-C6烷基、C1-C6卤代烷基、C1-C6烷氧基或C1-C6卤代烷氧基；
R1为或NR1aR1b；
R1a和R1b独立地为H或C1-C6烷基；
带“*”碳原子表示当为手性碳原子时，为R构型、S构型或者两者的混合；
带“#”碳原子表示当为手性碳原子时，为R构型、S构型或者两者的混合；
带“&”碳原子表示当为手性碳原子时，独立地为R构型、S构型或者两者的混合；
如式I所示化合物不为以下化合物：
及其异构体。
在本发明某些优选实施方案中，所述的化合物或其药学上可接受的盐中的某些基团如下定义，未提及的基团同本发明任一方案所述(简称“在一些实施方案中”)，R4b中，所述卤素独立地为F、Cl、Br或I；例如Br。
在一些实施方案中，R4b中，所述C1-C6卤代烷基独立地为CHF2、CH2F或CF3。
在一些实施方案中，R7a和R7b中，所述卤素独立地为F、Cl、Br或I；优选为F。
在一些实施方案中，R7a和R7b中，所述C1-C6烷基独立地为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基。
在一些实施方案中，R7a和R7b中，所述C1-C6卤代烷基独立地为CHF2、CH2F或CF3，例如CHF2。
在一些实施方案中，环A和环C中，所述C6-C10芳基独立地为苯基或萘基，优选为苯基。
在一些实施方案中，环A和环C中，所述“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个、2个或3个的5-10元杂芳基”独立地为“杂原子选自N、O和S中的1种、2种或3种， 杂原子数为1个或2个的5-6元或9-10杂芳基”，例如为吡啶基(例如)。
在一些实施方案中，环A和环C中，所述C3-C6环烷基独立地为环丙基、环丁基、环戊基或环己基。
在一些实施方案中，RA和RC中，所述卤素独立地为F、Cl、Br或I；优选为F或Cl，例如F。
在一些实施方案中，RA和RC中，所述C1-C6烷基独立地为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基。
在一些实施方案中，RA和RC中，所述C1-C6卤代烷基独立地为CHF2、CH2F或CF3。
在一些实施方案中，RA和RC中，所述C1-C6烷氧基独立地为甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、异丁氧基、仲丁氧基或叔丁氧基，优选为甲氧基或乙氧基。
在一些实施方案中，RA和RC中，所述C1-C6卤代烷氧基独立地为-OCHF2、-OCH2F或-OCF3；优选为-OCF3。
在一些实施方案中，R1a和R1b中，所述C1-C6烷基独立地为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基，例如甲基。
在一些实施方案中，式I中，带“*”碳原子表示当为手性碳原子时，为S构型。
在一些实施方案中，式I中，为
在一些实施方案中，R4a为H；R4b为卤素(例如Br)、CN、-CH2-CN、-CH2-OH、-CH(CH3)-OH、-COOH、-CH2-COOH、-CH2-OAc或NH2；或者，R4a和R4b以及它们连接的碳原子一起共同形成 例如，R4a为H；R4b为Br、CN、-CH2-CN、-CH2-OH、-CH(CH3)-OH、-COOH、-CH2- OAc或NH2；或者，R4a和R4b以及它们连接的碳原子一起共同形成
在一些实施方案中，当7号碳原子和8号碳原子之间为单键时，R7a和R7b独立地为H或卤素(例如F)，较佳的，R7a和R7b同时为H或卤素(例如F)；
当7号碳原子和8号碳原子之间为双键时，R7a不存在，R7b为卤素(例如F)；R8a不存在。
在一些实施方案中，R21为例如
在一些实施方案中，环A和环C独立地为C6-C10芳基或“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个、2个或3个的5-10元杂芳基”。
在一些实施方案中，RA和RC独立地为卤素、C1-C6烷氧基或C1-C6卤代烷氧基。
在一些实施方案中，n1为3。
在一些实施方案中，n3为3。
在一些实施方案中，m1为0、1、2或3。
在一些实施方案中，m3为0、1、2或3。
在一些实施方案中，为
在一些实施方案中，为
在一些实施方案中，为
在一些实施方案中，R21为
本发明还提供了一种式II所示化合物或其药学上可接受的盐：
其中，
R4c为H；R4d为H或OH；
R7c为H，R7d独立地为CN、-CH2-OH、-COOH、C1-C6烷基、C1-C6卤代烷基、-CH2-COOH、NH2、-CH2-CN或Cl；或者，R7c和R7d以及它们连接的碳原子一起共同形成
R22为
n2为2、3、4或5；
m2为0、1、2、3、4或5；
环B为C6-C10芳基、“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个、2个或3个的5-10元杂芳基”或C3-C6环烷基；
RB独立地为卤素、C1-C6烷基、C1-C6卤代烷基、C1-C6烷氧基或C1-C6卤代烷氧基；
带“*”碳原子表示当为手性碳原子时，为R构型、S构型或者两者的混合；
带“#”碳原子表示当为手性碳原子时，为R构型、S构型或者两者的混合；
带“&”碳原子表示当为手性碳原子时，为R构型、S构型或者两者的混合；
如式II所示化合物不为以下化合物：
及其异构体。
在一些实施方案中，R7c和R7d中，所述C1-C6烷基独立地为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基。
在一些实施方案中，R7c和R7d中，所述C1-C6卤代烷基独立地为CHF2、CH2F或CF3，例如CHF2。
在一些实施方案中，环B中，所述C6-C10芳基为苯基或萘基，优选为苯基。
在一些实施方案中，环B中，所述“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个、2个或3个的5-10元杂芳基”为“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个或2个的5-6元或9-10杂芳基”，例如为吡啶基(例如)。
在一些实施方案中，环B中，所述C3-C6环烷基为环丙基、环丁基、环戊基或环己基。
在一些实施方案中，RB中，所述卤素独立地为F、Cl、Br或I；优选为F或Cl，例如F。
在一些实施方案中，RB中，所述C1-C6烷基独立地为甲基、乙基、正丙基、异丙基、正丁基、异 丁基、仲丁基或叔丁基。
在一些实施方案中，RB中，所述C1-C6卤代烷基独立地为CHF2、CH2F或CF3。
在一些实施方案中，RB中，所述C1-C6烷氧基独立地为甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、异丁氧基、仲丁氧基或叔丁氧基，优选为甲氧基或乙氧基。
在一些实施方案中，RB中，所述C1-C6卤代烷氧基独立地为-OCHF2、-OCH2F或-OCF3；优选为-OCF3。
在一些实施方案中，式II中，带“*”碳原子表示当为手性碳原子时，为S构型。
在一些实施方案中，式II中，带“#”碳原子表示当为手性碳原子时，为R构型。
在一些实施方案中，式II中，为
在一些实施方案中，R7c为H，R7d独立地为CN、-CH3、-CH2-OH、-COOH、-CHF2、-CH2-COOH、NH2、-CH2-CN或Cl；或者，R7c和R7d以及它们连接的碳原子一起共同形成 例如，R7c为H，R7d独立地为CN、-CH2-OH、-CH3、NH2、-CH2-CN、-CF2H、-COOH，或者，R7c和R7d以及它们连接的碳原子一起共同形成
在一些实施方案中，R22为
在一些实施方案中，环B为C6-C10芳基或“杂原子选自N、O和S中的1种、2种或3种，杂原 子数为1个、2个或3个的5-10元杂芳基”。
在一些实施方案中，RB独立地为卤素、C1-C6烷氧基或C1-C6卤代烷氧基。
在一些实施方案中，n2为3。
在一些实施方案中，m2为0、1、2或3。
在一些实施方案中，为
在一些实施方案中，R22为
在一些实施方案中，式II中，为
本发明还提供了一种式III所示化合物或其药学上可接受的盐：
其中，
R3a为H或C1-C6烷基；
R3b为-O-(CH2)n-R3-1、NR3-2R3-3或OH；当R3b为OH时，R3a为C1-C6烷基；
n为1、2、3或4；R3-1为OH或-C(O)O-R3-1a；R3-1a为H或C1-C6烷基；
R3-2和R3-3独立地为H或-S(O)2R3-2a；R3-2a为C1-C6卤代烷基或C1-C6烷基；
R4e为H；R4f为H、OH或-O-(CH2)m-O-R4-1；或者R4e和R4f以及它们连接的碳原子一起共同形成m为1或2；R4-1为C1-C6烷基；
带“*”碳原子表示当为手性碳原子时，为R构型、S构型或者两者的混合；
带“#”碳原子表示当为手性碳原子时，为R构型、S构型或者两者的混合；
如式III所示化合物不为以下化合物：
及其异构体。
在一些实施方案中，R3a中，所述C1-C6烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基，例如甲基。
在一些实施方案中，R3-1a中，所述C1-C6烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基，例如叔丁基。
在一些实施方案中，R3-2a中，所述C1-C6卤代烷基为CHF2、CH2F或CF3，例如CF3。
在一些实施方案中，R4-1中，所述C1-C6烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基，例如乙基。
在一些实施方案中，式III中，为
在一些实施方案中，n为1、2或3。
在一些实施方案中，R3-2为H，R3-3为-S(O)2R3-2a。
在一些实施方案中，m为1。
在一些实施方案中，R3a为H，R3b为 或者，R3a为甲基，R3b为OH。
在一些实施方案中，R4e为H；R4f为H、OH或或者R4e和R4f以及它们连接的碳原子一起共同形成
在一些实施方案中，式III中，为
本发明还提供了如下任一所示的化合物或其药学上可接受的盐：








本发明还提供了一种药物组合物，其包含本发明任一项所述的化合物或其药学上可接受的盐，以及至少一种药用辅料。
本发明还提供了一种如本发明任一项所述的化合物或其药学上可接受的盐或如本发明任一项所述的药物组合物在制备用于预防和/或治疗疾病的药物中的应用，其中所述疾病为肥胖、高脂血症、脂肪肝、糖尿病、动脉粥样硬化症、心脑血管疾病、肝癌或皮肤损伤。
本发明还提供了一种如本发明任一项所述的化合物或其药学上可接受的盐或如上所述的药物组合物在制备用于抑制SREBP通路的药物中的应用。
本发明还提供了一种抑制SREBP通路的方法，其包括给予受试者有效量的如上所述的化合物或其药学上可接受的盐。
本发明还提供了一种预防和/或治疗疾病的方法，其包括给予受试者有效量的如本发明任一项所述的化合物或其药学上可接受的盐，其中所述疾病为肥胖、高脂血症、脂肪肝、糖尿病、动脉粥样硬化症、心脑血管疾病、肝癌或皮肤损伤。
定义和说明
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。
在本文中，术语“取代”或“取代基”是基团中的氢原子被指定的基团所代替。当没有指明取代位置时，取代可以在任何位置，但是只有形成一个稳定的或者是化学意义上可行的化学物才是被允许的。举例说明如下：结构表示环A上的氢原子被m1个RA所取代，当存在多个RA时，每个RA相同或不同。
当任何变量(例如RA)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。
在本文中，术语“烷基”是指饱和的直链或支链的一价烃基。C1-C6烷基表示具有1-6个碳原子的烷基。在一些实施方案中，C1-C6烷基可以为C1-C4烷基。C1-C4烷基包括甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基。
在本文中，术语“卤代烷基”是指烷基中的一个或多个氢原子被卤素所取代形成的基团，其中烷基的定义如前所述。卤代烷基的实例包括但不限于一氟甲基、二氟甲基、三氟甲基、五氟乙基等。
在本文中，术语“烷氧基”是指-O-烷基，其中烷基的定义如前所述。C1-C4烷氧基是指-O-(C1-C4烷基)，其中C1-C4烷基的定义如前所述，也即C1-C4烷氧基具体可以为甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、异丁氧基、仲丁氧基或叔丁氧基。
在本文中，术语“环烷基”是指饱和的单环或多环(例如并环、螺环或桥环)的环状烃基。环烷基的实例包括但不限于环丙基、环丁基、环戊基、环己基、等。C3-6环烷基具体可以为C3、C4、C5、C6环烷基。在一些实施方案中，环烷基为单环的。在一些实施方案中，环烷基为多环的(例如并环、螺环或桥环)。
在本文中，术语“C6-10芳基”是指苯基或萘基。
在本文中，术语“杂芳基”是指由碳原子和至少一个杂原子形成的芳香性的单环或稠环基团，其中杂原子独立地选自N、O和S中的1种、2种或3种。5-10元杂芳基具体可以为5、6、7、8、9或10元杂芳基，例如5-6元杂芳基或8-10元稠杂芳基。5-6元杂芳基是单环的，具体实例包括但不限于吡咯、呋喃、噻吩、噁唑、异噁唑、噻唑、异噻唑、吡唑、咪唑、吡啶、嘧啶、吡嗪。8-10元稠杂芳基的实例包括但不限于苯并吡咯、苯并呋喃、苯并噻吩、苯并噁唑、苯并异噁唑、苯并噻唑、苯并异噻唑、苯并吡唑、苯并咪唑、苯并吡啶、苯并嘧啶、苯并吡嗪、噻唑并噻唑、吡啶并吡啶、吡啶并吡嗪、 吡啶并嘧啶、
在本文中，化学结构式中的表示连接位置。当包含于环状基团中并且没有指明所连接的环原子时，可以连接于任何环原子，但是只有形成一个稳定的或者是化学意义上可行的化学物才是被允许的。
在本文中，术语“药学上可接受的盐”表示由适宜的非毒性有机酸、无机酸、有机碱或无机碱与化合物形成的盐，其保留化合物的生物活性。所述的有机酸可为本领域常规的能成盐的各种有机酸。所述的无机酸可为本领域常规的能成盐的各种无机酸。所述的有机碱可为本领域常规的能成盐的各种有机碱。所述的无机碱可为本领域常规的能成盐的各种无机碱。
化学结构中，用楔形实线键和楔形虚线键表示一个立体中心的绝对构型，用直形实线键和直形虚线键表示立体中心的相对构型。键并未指定构型，即如果化学结构中存在构型异构，键可以为或者同时包含两种构型(例如 的比例为1:1)。碳碳双键并未指明其具体构型时，其可以为E或Z构型。立体异构体可以使用手性原料合成、手性拆分制备或者可以使用常规技术例如但不限于使用手性柱的高效液相(HPLC)拆分。
在本文中，术语“受试者”包括任何动物，优选哺乳动物，更优选人。
在本文中，术语“有效量”是指无毒的但能达到预期效果的药物或药剂的足够用量。有效量的确定因人而异，取决于受体的年龄和一般情况，也取决于具体的活性物质，个案中合适的有效量可以由本领域技术人员根据常规试验确定。
在不违背本领域常识的基础上，上述各优选条件，可任意组合，即得本发明各较佳实例。
本发明所用试剂和原料均市售可得。
本发明的积极进步效果在于：本发明提供了一类新化合物，其对SREBP通路具有抑制活性，可用于预防和/或治疗肥胖、高脂血症、脂肪肝、糖尿病、动脉粥样硬化症、心脑血管疾病、肝癌、皮肤损伤等疾病。
具体实施方式
下面通过实施例的方式进一步说明本发明，但并不因此将本发明限制在所述的实施例范围之中。下列实施例中未注明具体条件的实验方法，按照常规方法和条件，或按照商品说明书选择。
关键中间体的制备
实施例I&II
中间体I(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-二氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己醛I
中间体II(5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-十四氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-8-基]己醛II的制备
第一步：将(22E,24S)-豆甾-6(5),22(23)-二烯-3β-醇I-1(5.00g,12.11mmol,1.0eq)溶解于二氯甲烷(50mL)中，将反应体系置于冰水浴中，冷却至5℃左右，依次向反应体系中加入乙酸酐(3.4mL,36.35mmol,3.0eq)，4-二甲氨基吡啶(300mg,2.42mmol,0.2eq)和三乙胺(8mL,60.57mmol,5.0eq)，将反应体系置于室温下搅拌2小时。TLC(石油醚:乙酸乙酯＝10:1)监测反应完全后，用甲醇(20mL)淬灭，反应液用饱和碳酸氢钠(～50mL)和水(～50mL)各洗涤一次，无水硫酸钠干燥，浓缩，在浓缩快干时，加入甲醇(～20mL)，冰浴下搅拌30分钟，抽滤，滤饼用少量甲醇淋洗，滤饼干燥得到白色固体乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E,5S)-5-乙基-6-甲基庚-3-烯-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-2(5.30g，纯度90.0％，收率86.58％)。1H NMR(400MHz,CDCl3)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),2.01–1.92(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).
第二步：称取乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E,5S)-5-乙基-6-甲基庚-3-烯-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-2(10.0g,22mmol,1eq)溶于四氢呋喃(200mL)和水(20.0mL)中，室温下加入吡啶(4.5mL,55mmol,2.5eq),N-甲基吗啉氧化物(10.30g,88mmol,4eq),锇酸钾(0.81g,2.2mmol,0.1eq),室温搅拌过夜，TLC(石油醚:乙酸乙酯＝3:1)监测，有部分原料剩余，有中间体(邻二醇生成)，随后在0℃下向该反应液中加入高碘酸钠(18.80g,88mmol,4eq),室温搅拌1小时，TLC(石油醚:乙酸乙酯＝3:1)检测，有部分原料剩余，中间体转化为产物。随后加入水50mL,乙酸乙酯50mL×3萃取，干燥，浓缩后粗品经柱层析(石油醚:乙酸乙酯＝60:1)纯化后得乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R,3E,5S)-5-乙基-6-甲基庚-3-烯-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-3(3.3g,纯度60％，收率19.8％)白色固体乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aS)-1-[(1S)-1-甲酰基乙基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯(1.9g,纯度90.0％，收率21.20％)。1H NMR(400MHz,CDCl3)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),1.89(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).
第三步：(甲氧基甲基)三苯基氯化磷(55.21g,161.052mmol,5.0eq)溶解在无水四氢呋喃(100mL)中，体系干冰乙酸乙酯浴降温到-10℃，加入二(三甲基硅基)氨基钠(80.526mL,1mol/L,2.5eq)，保持干冰乙酸乙酯浴搅拌30分钟后，乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aS)-1-[(1S)-1-甲酰基乙基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-3(12.0g,13.4mmol，1.0eq)溶解在无水四氢呋喃(50mL)中，加入到反应液中，体系恢复至室温，搅拌30分钟。TLC(石油醚:乙 酸乙酯＝10:1)监测反应完全。100mL饱和碳酸氢钠溶液缓慢加入到反应体系中，乙酸乙酯(100mL×3)萃取，收集有机相用水(100mL×2)洗，饱和盐水洗涤，无水硫酸钠干燥，有机相浓缩得到黄色固体乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2S,3E)-4-甲氧基丁-3-烯-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-4(8g,纯度90.0％，收率65.0％)，粗品直接投下一步反应。
第四步：乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2S,3E)-4-甲氧基丁-3-烯-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-4(8g,12.5mmol,1.0eq)溶于四氢呋喃(100mL)中，缓慢加入稀盐酸(5mol/L,50mL)，反应混合物室温搅拌30分钟。TLC(石油醚:乙酸乙酯＝10:1)监测反应。反应完全后，80mL水加入到反应中，反应液用乙酸乙酯(50mL×3)萃取，饱和盐水(20mL×3)洗涤，无水硫酸干燥，收集有机相浓缩得到粗品。粗品通过硅胶柱层析纯化(石油醚:乙酸乙酯＝50:1to 30:1to 4：1)浓缩得白色固体乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-甲酰基丙-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-5(6.3g,纯度90％，收率48.63％)。1H NMR(400MHz,CDCl3)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).
第五步：(将反应物乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-甲酰基丙-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-5(10.00g,25.87mmol，1.0eq)溶解在无水四氢呋喃(150mL)中，加入(三苯基-λ5-甲磷亚基)乙酸甲酯(51.89g,155.21mmol,6.0eq)，反应体系在90℃搅拌18h。核磁监测反应原料消耗完。100mL水加入到反应体系中，使用乙酸乙酯(100mL×3)萃取，收集有机相用水(100mL×2)洗，饱和盐水洗涤，无水硫酸钠干燥，收集有机相浓缩得到粗品。粗品通过柱层析(石油醚:乙酸乙酯＝60：1to 30：1)纯化，得到白色固体(2E,5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己-2-烯酸甲酯I-6(9.0g,纯度90％，收率71.1％)。1H NMR(400MHz,CDCl3)δ6.95(ddd,J＝15.4,8.7,6.4Hz,1H),6.26(d,J＝11.6Hz,0H),5.82(d,J＝15.5Hz,1H),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),1.68–1.40(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).
第六步：将反应物(2E,5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己-2-烯酸甲酯I-6(10g,22.59mmol，1.0eq)溶于四氢呋喃(100mL)和甲醇(50mL)混合溶剂中，加入氯化镍(2.93g,22.59mmol，1.0eq)，缓慢加入硼氢化钠(1.28g,33.89mmol)，反应体系在室温搅拌1h。核磁监测，反应 原料消耗完。100mL水加入到反应体系中，使用乙酸乙酯(100mL×3)萃取，收集有机相用水(100mL×2)洗，饱和盐水洗涤，无水硫酸钠干燥，收集有机相浓缩得到粗品。粗品通过柱层析(石油醚:乙酸乙酯＝60：1to 30：1)纯化，得到白色固体(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-7(9g,纯度90％，收率80.6％)。1H NMR(400MHz,CDCl3)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).
第七步：室温下在250mL圆底烧瓶中，将(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-7(2.5g,5.62mmol)溶解于三氯甲烷(80mL)，在室温下加入N-甲基吗啉(1.71g,16.87mmol)和二氧化硒(1.56g，14.06mmol)，随后在70℃下搅拌18小时。TLC(石油醚:乙酸乙酯＝10:1)监测反应。反应结束后，用水(100mL)淬灭，二氯甲烷(80mL×3)萃取，有机相饱和盐水(150mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)，得到白色固体(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-8(1.4g，纯度:90％，产率：48.65％)。1H NMR(400MHz,CDCl3)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).
第八步：将反应物(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-8(5g,11.5mmol，1.0eq)溶于甲醇(150mL)中，加入碳酸钾(6.37g,46mmol，4.0eq)，反应体系在室温搅拌30min。TLC(石油醚:乙酸乙酯＝5:1)监测反应。原料消耗完毕，停止反应。将水(100mL)加入到反应体系中，水层用乙酸乙酯(3×50mL)萃取。合并乙酸乙酯层并且用饱和食盐水(3×50mL)洗涤。乙酸乙酯层用无水硫酸钠干燥，过滤和浓缩得到粗品(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-9，粗品直接用于下一步反应。
第九步：将反应物(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-9(5g,11.9mmol,1eq)溶于丙酮(150mL)中，加入对甲苯磺酸(1.59g,8.4mmol,0.7eq)和4A分子筛，反应体系室温搅拌1h。TLC(石油醚:乙酸乙酯＝10:1)监测反应。原料消耗完毕，用饱和亚硫酸钠淬灭停止反应。将100mL水加入到反应体系中，使用乙酸乙酯(50mL×3)萃取，收集有机相，使用无水硫酸钠干燥，有机相真空旋干得到粗品。粗品溶于乙酸乙酯中通过柱层析(石油醚:乙酸乙酯＝95:5)得到白色固体 (5R)-5-[(3aR,3R,5aS,9aS,9bS)-7-[(4R)-2,2-二甲基-1,3-二氧杂环戊-4-基]-3a,6,6-三甲基-2,3,3a,4,5,5a,6,9,9a,9b-十氢-1H-环戊并[1,2-a]萘-3-基]己酸甲酯I-10(3.8g,纯度90％，收率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).
第10步：将化合物(5R)-5-[(3aR,3R,5aS,9aS,9bS)-7-[(4R)-2,2-二甲基-1,3-二氧杂环戊-4-基]-3a,6,6-三甲基-2,3,3a,4,5,5a,6,9,9a,9b-十氢-1H-环戊并[1,2-a]萘-3-基]己酸甲酯I-10(3.8g,8.2mmol,1.0eq)溶解在丙酮(50mL)中,加入N-羟基邻苯二甲酰亚胺(0.54g,3.3mmol,0.8eq),叔丁基过氧化氢(12mL,66mmol,8.0eq)和无水醋酸钴(II)(0.29g,1.6mmol,0.2eq)，所得混合液在反应液在N2，25℃中搅拌18小时。TLC(石油醚:乙酸乙酯＝10:1)监测反应。原料基本消耗完毕，用饱和亚硫酸钠淬灭,将水(10mL)加入到反应体系中，水层用乙酸乙酯(3×15mL)萃取。合并有机层并且用饱和食盐水(15mL)洗涤，无水硫酸钠干燥，过滤和浓缩得到粗品。粗品硅胶柱层析纯化(石油醚:乙酸乙酯＝87：13)，得到白色固体(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-四甲基-11-氧亚基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-十四氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己酸甲酯I-11(2.5g,纯度95％.收率60％)。1H NMR(400MHz,CDCl3)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).
第11步：将化合物(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-11(2.4g,5.1mmol，1.0eq)溶解在甲醇(100mL)和乙酸乙酯(50mL)中，然后加入钯碳(1.2g,11mmol，2.2eq)并将反应体系置换成氢气氛围，将反应体系置于室温搅拌1h。TLC(石油醚:乙酸乙酯＝5:1)监测反应，原料消耗完毕。过滤反应体系，收集有机相并浓缩，得到粗品。粗品通过硅胶柱层析(石油醚:乙酸乙酯＝90:10)得到白色固体(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-11-氧亚基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己酸甲酯I-12(1.7g,纯度95％，收率67％)。1H NMR(400MHz,CDCl3)δ4.00–3.87(m,2H),3.60(s,3H),2.75–2.66(m,1H),2.36(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).
第12步将化合物(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-11-氧亚基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳 -8-基]己酸甲酯I-12(1.7g,3.6mmol)溶解在二乙胺基三氟化硫(10mL)中,所得混合液在反应液在80℃中搅拌1h。TLC板(石油醚:乙酸乙酯＝10:1)监测反应完全。反应液降至室温，加入二氯甲烷(50mL)稀释反应体系，小心的滴加冰水淬灭反应，分层收集有机相，水层用二氯甲烷(3×30mL)萃取，合并有机相用饱和食盐水(40mL)洗涤，过滤和浓缩得到粗品。粗品加入硅胶过柱(石油醚：乙酸乙酯＝93：7)，得到无色透明固体(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-二氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己酸甲酯I-13和(5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-十四氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-8-基]己酸甲酯II-1的混合物(1.4g，I-13纯度95％,以I-13计算收率:74％)。
化合物I-13：1H NMR(400MHz,CDCl3)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),1.46–1.34(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).
第十三步将化合物(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-二氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己酸甲酯I-13和(5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-十四氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-8-基]己酸甲酯II-1的混合物(1.3g,2.6mmol,1.0eq)溶解在四氢呋喃(50mL)中,加入四氢铝锂(0.3g,7.8mmol,3eq)，所得混合液在反应液在25℃中搅拌1h。TLC(石油醚:乙酸乙酯＝10:1)监测反应完全。用十水合硫酸钠淬灭，过滤和浓缩得到白色固体(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-二氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己-1-醇I-14和(5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-十四氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-8-基]己酸甲酯II-12粗品(0.9g，纯度96％,收率:92％)，直接投下一步。
第十四步：将化合物(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-二氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己-1-醇I-14和(5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-十四氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-8-基]己酸甲酯II-2混合物(1.3g,2.7mmol,1.0eq)溶解在二氯甲烷(50mL)中,加入戴斯马丁试剂(2.3g,5.5mmol,2eq)，所得混合液在反应液在N2，25℃中搅拌1h。TLC(石油醚:乙酸乙酯＝10:1)监测反应完全。将饱和碳酸氢钠(10mL)加入到反应体系中，有机相用饱和亚硫酸钠(3×30mL)萃取， 收集浓缩有机相。硅胶柱层析纯化(石油醚:乙酸乙酯＝92：8)，得到白色固体(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-二氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己醛I和(5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-十四氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-8-基]己醛II混合物(0.95g,I的纯度90％，以I计算收率66％)。
化合物I：1H NMR(400MHz,CDCl3)δ9.70(t,J＝1.7Hz,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＝10.7,10.3,4.7Hz,1H),1.57–1.52(m,1H),1.44(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)。
实施例III
中间体III(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-十四氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己醛(III)的制备
第一步：将化合物(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-十四氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己酸甲酯I-10(1g,2.18mmol,1.0eq)溶解在四氢呋喃(50mL)中,氮气置换后加入四氢铝锂(0.12g,3.270mmol,1.5eq)，25℃中搅拌1h。TLC板(石油醚:乙酸乙酯＝5:1)监测反应进度，原料消耗完毕。用十水合硫酸钠淬灭,过滤，滤液浓缩得到粗品。粗品硅胶柱层析纯化(石油醚:乙酸乙酯＝80:20)得到白色固体(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-十四氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己-1-醇III-1(1g,纯度:90％,收率:96％).1H NMR(400MHz,CDCL3)δ5.80(d,J＝2.6Hz,1H),4.41(d,J ＝5.8Hz,1H),4.09(m,1H),3.64(t,J＝6.5Hz,2H),2.08(m,2H),1.84(tt,J＝18.5,7.8Hz,2H),1.67(m,8H),1.41(td,J＝12.7,6.2Hz,4H),1.35(s,3H),1.24(m,2H),1.17(s,4H),1.08(m,5H),0.93(d,J＝6.5Hz,4H),0.69(d,J＝4.6Hz,3H).
第二步：将化合物(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-十四氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己-1-醇III-1(1g,90％纯度，2.09mmol,)溶解在二氯甲烷(50mL)中,加入戴斯马丁氧化剂(1.18g,2.79mmol)，25℃中搅拌1h。TLC(石油醚:乙酸乙酯＝5:1)监测反应进度，原料消耗。用饱和亚硫酸钠淬灭,将水(10mL)加入到反应体系中，水层用二氯甲烷(3×50mL)萃取。合并有机相并且用饱和食盐水(10mL)洗涤，无水Na2SO4干燥，过滤，浓缩得到粗品。粗品硅胶柱层析纯化(石油醚：乙酸乙酯＝90：10)得到白色固体(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-十四氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己醛III(750mg,1.58mmol,纯度：90％，收率：68％)。1H 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).
实施例9&10
化合物9 3β-[(2-羟基乙基)氧基]-5α-胆甾-25-醇和
化合物10{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯

第一步：将乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-甲氧基-6-氧亚基己-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-7(80mg,0.180mmol)溶于四氢呋喃(2mL)和甲醇(1mL)中，加入Pd(OH)2(20mg,0.142mmol)，氢气置换三次，40℃搅拌48小时，HNMR监测。反应液过滤，滤液浓缩得乙酸-(1R,3aS,3bR,7S,9aS,9bS,11aR)-1-[(2R)-6-甲氧基-6-氧亚基己-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯9-1(80mg,0.170mmol,94.57％)类白色固体。1HNMR(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),1.48–1.40(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).
第二步：将甲基乙酸-(1R,3aS,3bR,7S,9aS,9bS,11aR)-1-[(2R)-6-甲氧基-6-氧亚基己-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯9-1(80mg,0.179mmol)溶于四氢呋喃(3mL)中，氮气置换三次，降温至-78℃，加入甲基锂(0.560mL,0.896mmol)后，升至室温搅拌3小时。TLC(石油醚:乙酸乙酯＝10:1)监测。反应液加水(15mL)淬灭，用乙酸乙酯(10mL*3)萃取，盐水洗涤一次，无水硫酸钠干燥，浓缩得到80mg粗品。柱层析纯(DCM：MeOH＝300:1-100：1)得到胆甾-3β,25-二醇9-2(20mg,0.040mmol,22.08％)。1H NMR(399MHz,CDCl3)δ5.32(s,0H),3.56(s,1H),1.93(d,J＝12.9Hz,1H),1.75(s,2H),1.70–1.60(m,2H),1.55(s,3H),1.43(s,3H),1.35(s,3H),1.28(s,3H),1.24(s,3H),1.23(s,3H),1.19(s,6H),1.11–1.01(m,4H),0.95(d,J＝10.9Hz,3H),0.89(d,J＝6.5Hz,3H),0.77(s,3H),0.62(s,3H).13CNMR(101MHz,CDCl3)δ77.31,77.20,77.00,76.68,71.35,71.10,56.45,56.16,54.30,44.81,44.39,42.58,40.00,38.19,36.97,36.39,35.73,35.47,35.43,32.06,31.50,30.95,29.33,29.18,28.71,28.25,27.19,24.19,21.23,20.74,18.61,12.31,12.06.LC-MS:[M+Na]+＝427.15.
第三步：将5α-胆甾-3β,25-二醇(10mg,0.025mmol)溶于甲苯(3mL)中，加入叔丁醇钾(14.03mg, 0.125mmol)，室温搅拌1小时。加入溴乙酸叔丁酯(24.38mg,0.125mmol)，室温搅拌2小时。TLC(石油醚：乙酸乙酯＝3:1)监测。反应液直接浓缩得到40mg粗品，柱层析纯化(乙酸乙酯：石油醚＝30％)得到{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯10(3mg,0.005mmol,22.23％)，白色固体。
化合物10：1HNMR(399MHz,Chloroform-d)δ3.98(s,2H),3.29(dt,J＝11.2,6.2Hz,1H),1.90(dd,J＝30.3,12.5Hz,2H),1.79–1.67(m,2H),1.62(d,J＝13.0Hz,4H),1.51(d,J＝7.2Hz,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).
第四步：将{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯10(20mg,0.039mmol)溶于四氢呋喃(5mL)中，加入四氢铝锂(4.39mg,0.116mmol)，室温搅拌1小时。TLC(石油醚：乙酸乙酯＝3:1)监测。反应液直接浓缩得到10mg粗品，柱层析纯化(乙酸乙酯：石油醚＝40％)得到3β-[(2-羟基乙基)氧基]-5α-胆甾-25-醇9(10mg,0.021mmol,54.92％)，白色固体。
化合物9：1H NMR(399MHz,Chloroform-d)δ3.69(t,J＝4.5Hz,2H),3.55(td,J＝4.5,2.2Hz,2H),3.29–3.20(m,1H),2.10–1.91(m,2H),1.87–1.75(m,2H),1.71(dd,J＝13.4,3.9Hz,1H),1.62(d,J＝3.3Hz,2H),1.52(s,1H),1.49–1.39(m,3H),1.37(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),1.01–0.91(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).13C NMR(101MHz,CDCl3)δ79.02,71.10,68.82,62.11,56.46,56.18,54.35,44.78,44.39,42.59,40.00,36.91,36.40,35.73,35.46,34.79,32.07,29.32,29.17,28.81,28.24,24.18,21.20,20.74,18.61,12.26,12.0.LCMS:[M+Na]+＝458.39.
实施例12
化合物12(5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-7-氨基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯

第一步：将乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-甲氧基-6-氧亚基己-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-7(1g,2.249mmol)，溶于入四氢呋喃(5mL)和甲醇(5mL)中。加入碳酸钾(3.11g,22.489mmol)。40℃搅拌5小时，TLC(石油醚：乙酸乙酯＝3：1)监测。反应液浓缩得到粗品5g。柱层析纯化(乙酸乙酯/石油醚＝40％)，得到(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯12-1(0.8g,1.888mmol,83.94％)，白色固体。1H 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).
第二步：室温下50mL圆底烧瓶中，将(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯12-1(250mg,0.62mmol)溶解于乙酸乙酯(10mL)，在室温下加入乙酸(0.2mL)和二氧化铂(125mg)，室温下将混合物在氢气氛围中搅拌16小时。TLC(乙酸乙酯/石油醚＝5:1)检测反应，有新的化合物生成。反应结束后，用乙酸乙酯(30mL)稀释反应液，通过硅藻土过滤，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝5:1)，得白色固体(5R)-5-[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-7-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯12-2(240mg，纯度：90％，产率：85.97％)。1H 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).
第三步：室温下50mL圆底烧瓶中，将(5R)-5-[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-7-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯12-2(230mg,0.57mmol)溶解于1，2-二氯乙烷(10mL)， 在室温下加入戴斯-马丁氧化剂(362mg，0.85mmol)，在45℃下搅拌3小时。TLC(乙酸乙酯/石油醚＝10：1)检测反应。反应结束后，用二氯甲烷(10mL)稀释反应液，加入饱和亚硫酸钠水溶液(20mL)和饱和碳酸氢钠水溶液(20mL)淬灭反应，二氯甲烷(20mL×3)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)，得白色固体(5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-9a,11a-二甲基-7-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯12-3(220mg，纯度：90％，产率：86.52％)。1H 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).13C NMR(101MHz,CDCl3)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.
第四步：室温下50mL圆底烧瓶中，将(5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-9a,11a-二甲基-7-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯12-3(190mg,0.47mmol)溶解于四氢呋喃(3mL)和甲醇(6mL)，在室温下加入醋酸铵(363mg，4.70mmol)和催化量的乙酸(0.20mL)，室温下搅拌2小时。随后向反应液中加入氰基硼氢化钠(297mg，4.70mmol)，室温下搅拌14小时。TLC(二氯甲烷/甲醇＝10：1)监测反应。反应结束后，用二氯甲烷(20mL)稀释反应液，加入饱和碳酸氢钠水溶液(30mL)淬灭反应，二氯甲烷(20mL×3)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(二氯甲烷：甲醇＝10：1)，得白色固体(5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-7-氨基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯12-4(160mg，纯度：90％，产率：75.59％)。1H 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).
第五步：室温下在50mL的圆底烧瓶中，将(5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-7-氨基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯12-4(160mg,0.396mmol)溶解于无水四氢呋喃(5mL)，反应氮气保护下，降温到0℃，滴加三摩尔每升的甲基溴化镁四氢呋喃溶液(1.321mL，3.963mmol)，随后转至室温搅拌2小时，TLC(二氯甲烷:甲醇＝8：1)监测反应，反应结束后，体系降温至0℃，用饱和氯化铵水溶液(20mL)淬灭，乙酸乙酯(15mL×3)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，硅胶柱层析纯化(二氯甲烷:甲醇＝8：1)，得到白色固体3-氨基-5α-胆甾-25-醇(130mg，纯度:80％，产率：64.99％)。室温下在50mL的圆底烧瓶中，将(5R)-5-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-7-氨基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯(160mg,0.396mmol)溶解于无水四氢呋喃(5mL)，反应氮气保护下，降温到0℃，滴加三摩尔每升的甲基溴化镁四氢呋喃溶液(1.321mL，3.963mmol)，随后转至室温搅拌2小时，TLC(二氯甲烷:甲醇＝8：1)监测反应，反应结束后，体系降温至0℃，用饱和氯化铵水溶液(20mL)淬灭，乙酸乙酯(15mL×3)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，硅胶柱层析纯化(二氯甲烷:甲醇＝8：1)，得到白色固体3-氨基-5α-胆甾-25- 醇12(130mg，纯度:80％，产率：64.99％)。(ESI-MS(M+H)+：404.4.1H NMR(400MHz,DMSO)δ7.61(s,2H),4.03(s,1H),2.93(m,1H),1.93(d,J＝12.0Hz,1H),1.70(s,4H),1.47(d,J＝13.4Hz,4H),1.31(dd,J＝13.5,6.9Hz,6H),1.18(m,10H),1.05(s,6H),0.99(m,4H),0.89(d,J＝6.5Hz,3H),0.82(d,J＝12.6Hz,1H),0.76(s,3H),0.63(s,3H).
实施例14
化合物14 1,1,1-三氟-N-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]甲烷磺酰胺的制备。
第一步：在50mL圆底烧瓶中，将3-氨基-5α-胆甾-25-醇12(20mg,0.050mmol)溶解于二氯甲烷(1mL)，在室温下加入三乙胺(15mg，0.15mmol)，在0℃下逐滴加入三氟甲磺酸酐(17mg，0.059mmol)，反应液在0℃下搅拌1小时。TLC(乙酸乙酯/石油醚＝3：1)监测反应。反应结束后，向反应液中加入水(10mL)，乙酸乙酯(5mL×3)萃取，有机相饱和盐水(10mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝3：1)，得白色固体1,1,1-三氟-N-[(1R,3aS,3bR,5aS,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]甲烷磺酰胺14(5mg，纯度：91.44％，产率：17.23％)。
化合物14：1H 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＝6.5Hz,3H),0.79(s,3H),0.65(s,3H).19F NMR(377MHz,CDCl3)δ-77.92(s,3F).
实施例18
化合物18 3β-[(3-羟基丙基)氧基]-5α-胆甾-25-醇的制备
第一步：将5α-胆甾-3β,25-二醇9-2(200mg,0.494mmol)溶于甲苯(10mL)中，加入叔丁醇钾(277.28mg,2.471mmol)，室温搅拌1小时。加入烯丙基溴(298.95mg,2.471mmol)，室温搅拌2小时。TLC(石油醚：乙酸乙酯＝3:1)监测。反应液直接浓缩得到40mg粗品，柱层析纯化(乙酸乙酯：石油醚＝30％)得到3β-(丙-2-烯基氧基)-5α-胆甾-25-醇18-1(70mg,0.150mmol,30.26％)，白色固体。1H 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).
第二步：将3β-(丙-2-烯基氧基)-5α-胆甾-25-醇18-1(60mg,0.135mmol)溶于四氢呋喃(3mL)中，加入硼烷四氢呋喃(1.349mL,1.349mmol)，室温搅拌1小时。加入氢氧化钠(0.899mL,2.698mmol)，双氧水(152.94mg,1.349mmol)，室温搅拌2小时。TLC(石油醚：乙酸乙酯＝1:1)监测。反应液直接浓缩得到400mg粗品，柱层析纯化(乙酸乙酯：石油醚＝50％)得到3β-[(3-羟基丙基)氧基]-5α-胆甾-25-醇18(20mg,0.041mmol,30.43％)，白色固体。
化合物18：1H NMR(400MHz,Chloroform-d)δ3.76(t,J＝5.4Hz,2H),3.65(tq,J＝5.5,3.2Hz,2H),3.21(dt,J＝11.1,6.0Hz,1H),1.94(dt,J＝12.8,3.3Hz,1H),1.86–1.82(m,1H),1.81–1.78(m,2H),1.77–1.66(m,3H),1.62(ddd,J＝12.6,6.5,3.1Hz,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).
.
实施例19
化合物19{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸的制备。
第一步：将{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯(140mg,0.270mmol)溶于二氯甲烷(3mL)中，加入甲酸(6mL,0.010mmol)，室温搅拌1小时。TLC(石油醚：乙酸乙酯＝3:1)监测。反应液直接浓缩得到粗品{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸19(140mg,0.121mmol,44.85％)，淡黄色固体。化合物19：1H 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＝20.6Hz,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.
实施例23
化合物23 3β,25-二羟基胆甾-4β-甲酸的制备
第一步室温下50mL圆底烧瓶中，将乙酸-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]甲基酯13(220mg,0.46mmol)溶解于二氯甲烷(10mL)，在室温下加入咪唑(157mg，2.31mmol)和叔丁基二甲基氯硅烷(348mg，2.31mmol)，随后在室温下搅拌16小时.TLC(石油醚:乙酸乙酯＝5:1)监测反应。反应结束后，用水(50mL)淬灭，乙酸乙酯(30mL×3)萃取，有机相饱和盐水(60mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝5：1)，得到白色固体乙酸-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]甲基酯23-1(180mg，纯度:90％，产率：59.40％)。1H 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).
第二步在50mL圆底烧瓶中，将乙酸-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]甲基酯23-1(140mg,0.24mmol)溶解于四氢呋喃(6mL)，在室温下加入1摩尔每升的四氢铝锂四氢呋喃溶液(0.28mL，0.28mmol)，在室温下搅拌0.5小时。TLC(乙酸乙酯/石油醚＝3：1)监测反应。反应结 束后，用水(50mL)淬灭，乙酸乙酯(30mL×3)萃取，有机相饱和盐水(50mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝3：1)，得白色固体3β-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-4-(羟基甲基)-5α-胆甾-25-醇23-2(100mg，纯度：90％，产率：69.21％)。1H 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),1.62–1.32(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).
第三步在50mL圆底烧瓶中，将3β-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-4-(羟基甲基)-5α-胆甾-25-醇23-2(50mg,0.091mmol)溶解于四氢呋喃(1.5mL)和二氯甲烷(1.5mL)，在室温下加入戴斯马丁试剂(58mg，0.14mmol)，在室温下搅拌2小时。TLC(乙酸乙酯/石油醚＝4：1)监测反应。反应结束后，用水(20mL)淬灭，乙酸乙酯(10mL×3)萃取，有机相饱和盐水(10mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，得白色固体3β-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-25-羟基-5α-胆甾-4-甲醛23-3(15mg，纯度：90％，产率：27.10％)。1H 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).
第四步室温下50mL圆底烧瓶中，将3β-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-25-羟基胆甾-4β-甲醛23-3(12mg,0.022mmol)溶解于叔丁醇(1mL)和四氢呋喃(0.5mL)，在0℃下加入2-甲基丁-2-烯(10mg，0.13mmol)，磷酸二氢钠(8mg,0.066mmol)，亚氯酸钠(7mg，0.077mmol)和水(0.25mL)，随后在室温下搅拌16小时。TLC(二氯甲烷:甲醇＝10:1)监测反应。反应结束后，用饱和亚硫酸钠水溶液(20mL)淬灭，二氯甲烷和甲醇(二氯甲烷:甲醇＝10:1，10mL×3)萃取，有机相饱和盐水(10mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(二氯甲烷:甲醇＝10：1)，得到白色固体3β,25-二羟基胆甾-4β-甲酸23(3.65mg，纯度:100％，产率：37.09％)。
化合物23：1H NMR(400MHz,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).13C 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-2H2O]+＝413.35
实施例24
化合物24 24,24-二氟胆甾-6(5)-烯-3β,25-二醇的制备。
第一步将锌(0.61g,9.312mmol)溶解于THF(15mL)中，然后氮气保护下回流搅拌1小时，乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-甲酰基丙-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-1-甲酰基丙-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-5(1.2g,3.104mmol)和BrCF2CO2Et(1.89g,9.312mmol)的混合物溶于THF(3mL)中，然后滴加到上述锌的溶液中，反应在回流下反应0.5小时，TLC(石油醚:乙酸乙酯＝5:1)监测原料反应完全。反应液倒入水中，乙酸乙酯萃取(20mL x 2)，有机相合并，浓缩，粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝95:5to 80:20)纯化，得到白色固体(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟-3-羟基己酸乙酯24-1(700mg,纯度:90％,收率：39.74％)。1H 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,3H),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).
第二步将(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟-3-羟基己酸乙酯24-1 (300mg,0.587mmol)溶于二氯甲烷(10mL)中,然后依次加入二(咪唑-1-基)甲硫酮(314.08mg,1.762mmol)和DMAP(14.35mg,0.117mmol)，反应在回流下反应12小时，TLC(石油醚:乙酸乙酯＝3:1)监测反应。反应液倒入水中，乙酸乙酯萃取(20mL x2)，有机相合并，浓缩，粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝95:5～60：40)纯化，得到无色油状液体(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟-3-{[(咪唑-1-基)硫亚基甲基]氧基}己酸乙酯24-2(250mg,纯度:90％,61.69％)。1H NMR(400MHz,CDCl3)δ8.47(d,J＝9.6Hz,1H),7.67(d,J＝11.0Hz,1H),7.14(d,J＝6.8Hz,1H),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).
第三步将(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟-3-{[(咪唑-1-基)硫亚基甲基]氧基}己酸乙酯24-2(200mg,0.322mmol)溶于甲苯(5mL)中,依次加入苯甲酸过氧酸酐(15.61mg,0.064mmol)和三乙基硅氢(299.69mg,2.577mmol)，反应在回流下反应18小时。TLC(石油醚:乙酸乙酯＝8:1)监测反应完全。反应液倒入水中，乙酸乙酯萃取(20mLx2)，有机相合并，浓缩，粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝95:5)纯化，得到(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟己酸乙酯24-3(90mg,纯度:90％,50.83％)，直接投下一步。
第四步将(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟己酸乙酯24-3(70mg,0.142mmol)溶解于THF(4mL)中，降温至00C,缓慢滴加3.0MMeMgBr(0.28mL)，加料完毕后在0℃下搅拌0.5小时。TLC(石油醚:乙酸乙酯＝5:1)监测反应完全后，用水(10mL)萃灭，乙酸乙酯萃取(10mLx2),浓缩，粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝95:5to 90:10)纯化，得到白色固体乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯24-4(50mg,90％,69.83％)。1H 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(dt,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).
第五步将乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯24-4(60mg,0.125mmol)溶于MeOH(5mL)和水(1mL)中,然后加入K2CO3(34.5mg,0.25mmol)，反应在室温反应1小时，TLC( 石油醚:乙酸乙酯＝3:1)监测有新点产生。反应液倒入水中，乙酸乙酯萃取(20mLx2)，有机相合并，浓缩，粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝95:5～60：40)纯化，得到白色固体24,24-二氟胆甾-6(5)-烯-3β,25-二醇24(30mg,0.065mmol,52.05％)。
化合物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
实施例25
化合物25 24,24-二氟-5α-胆甾-3β,25-二醇的制备。
第一步将得到白色固体乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯24-4(40mg,0.083mmol)溶解于甲醇(4mL)中，加入Pd/C(40mg)加料完毕后在室温下搅拌3小时。TLC(石油醚:乙酸乙酯＝2:1)监测反应完全后，过滤,乙酸-(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯25-1(40mg,90％,89.62％),粗品直接投下一步.
第二步将乙酸-(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯25-1(40mg,0.083mmol)溶于甲醇(5ml)和水(0.5ml)中,然后加入碳酸钾(34.36mg,0.249mmol)，反应在室温反应1小时，TLC(石油醚:乙酸乙酯＝3:1)监测有新点产生。反应液倒入水中，乙酸乙酯萃取(20mlx2)，有机相合并，浓缩，粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝95:5～60：40)纯化，得到白色固体24,24-二氟-5α-胆甾-3β,25-二醇25(10mg,0.020mmol,24.65％)。化合物25：1H 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
实施例26
化合物26 4β-羟基-3β-羟基-24-(羟基环丙基)-5α-胆烷-7-酮的制备。
第一步:将(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-8(300mg,0.65mmol)溶解在中二氯甲烷(20mL),加入4-二甲氨基吡啶(39.81mg,0.03mmol)，三乙胺(197.70mg,1.95mmol)和乙酸酐(79.78mg,0.78mmol)室温反应2h。TLC(石油醚：乙酸乙酯＝5:1)监测反应完全。用二氯甲烷和水洗萃后用无水硫酸钠干燥浓缩，柱层析(石油醚：乙酸乙酯～20:1)纯化得(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢- 1H-环戊并[1,2-a]菲-1-基]己酸甲酯26-1(230mg,0.441mmol,62.2％)白色固体。1H 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).
第二步:将(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯26-1(100.00mg,0.23mmol)溶解在丙酮(3.0mL)，氮气保护，N-羟基邻苯二甲酰亚胺(3.25mg,0.02mmol)，醋酸钴(0.35mg,0.01mmol)然后加入过氧叔丁醇(85mg,0.90mmol)，室温反应16h。TLC(石油醚：乙酸乙酯＝10:1)监测反应完全。加入硫代硫酸钠淬灭，乙酸乙酯萃取，无水硫酸钠干燥，浓缩，柱层析石油醚：乙酸乙酯(100％～20％),得到(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-9a,11a-二甲基-4-氧亚基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯26-2(40mg,0.07mmol,35.0％)。1H 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),2.00(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).
第三步:将(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-9a,11a-二甲基-4-氧亚基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯26-2(300mg,0.65mmol)溶解在甲醇(10.0mL)，Pd/C 10％(20mg,0.19mmol)，氢气换气三次，室温反应1h，TLC(石油醚：乙酸乙酯＝5:1)监测反应完毕。硅藻土过滤，甲醇漂洗，浓缩，柱层析石油醚：乙酸乙酯(100％～20％),得到(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-9a,11a-二甲基-4-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯26-3(50mg)白色固体，直接投下一步。1H 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＝3.7Hz,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).
第四步:将(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-9a,11a-二甲基-4-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯26-3(50mg,0.10mmol)溶解在甲苯(5mL)，加入乙二醇(0.05mL,0.96mmol)，无水对甲苯甲酸(7.3mg,0.05mmol)，加热回流搅拌1h。TLC(石油醚：乙酸乙酯＝2:1)监测反应。加入水，用乙酸乙酯萃取两次，无水硫酸钠干燥，浓缩柱层析石油醚：乙酸乙酯(100％～20％),得到(5R)-5-[(1'R,3a'S,3b'S,6'R,7'S,9a'R,9b'S,11a'R)-6',7'-二乙酰氧基-9a',11a'-二甲基-1',2',3',3a',3b',5',5a',6',7',8',9',9a',9b',10',11',11a'-十六氢螺[1,3-二氧杂环戊烷-2,4'-环戊并[1,2-a] 菲]-1'-基]己酸甲酯26-4(8mg,0.01mmol,17.2％)白色固体。1H NMR(399MHz,Chloroform-d)δ5.14(s,1H),4.76(dt,J＝12.0,4.3Hz,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＝6.5Hz,3H),0.62(s,3H)
第五步:将化合物(5R)-5-[(1'R,3a'S,3b'S,6'R,7'S,9a'R,9b'S,11a'R)-6',7'-二乙酰氧基-9a',11a'-二甲基-1',2',3',3a',3b',5',5a',6',7',8',9',9a',9b',10',11',11a'-十六氢螺[1,3-二氧杂环戊烷-2,4'-环戊并[1,2-a]菲]-1'-基]己酸甲酯26-4(20mg,0.04mmol)溶解在四氢呋喃(5mL)，室温加入钛酸四异丙脂(95.6mg,0.67mmol)，氮气换气三次，降温至0℃滴加乙基氯化镁(0.62mL)。室温搅拌1h，TLC(石油醚：乙酸乙酯＝3:1)监测。低温加入食盐水淬灭，加入乙酸乙酯稀释，无水硫酸钠干燥，浓缩，柱层析石油醚：乙酸乙酯(100％-50％)得(1'R,3a'S,3b'S,5a'R,6'R,7'S,9a'R,9b'S,11a'R)-1'-[(2R)-5-(羟基环丙基)戊-2-基]-9a',11a'-二甲基-1',2',3',3a',3b',5',5a',6',7',8',9',9a',9b',10',11',11a'-十六氢螺[1,3-二氧杂环戊烷-2,4'-环戊并[1,2-a]菲]-6',7'-二醇26-5(15.0mg,0.03mmol,33.6％)白色固体。1H 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),1.44(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)
第五步:将(1'R,3a'S,3b'S,5a'R,6'R,7'S,9a'R,9b'S,11a'R)-1'-[(2R)-5-(羟基环丙基)戊-2-基]-9a',11a'-二甲基-1',2',3',3a',3b',5',5a',6',7',8',9',9a',9b',10',11',11a'-十六氢螺[1,3-二氧杂环戊烷-2,4'-环戊并[1,2-a]菲]-6',7'-二醇26-5(10mg,0.02mmol)溶解甲醇(3.0mL)，室温加入醋酸(2mL,34.94mmol)，室温搅拌1h，TLC(石油醚：乙酸乙酯＝1:1)监测反应完全。低温加入食盐水淬灭，加入乙酸乙酯稀释，无水硫酸钠干燥，浓缩，柱层析石油醚：乙酸乙酯(100％-50％)得4β-羟基-3β-羟基-24-(羟基环丙基)-5α-胆烷-7-酮26(3mg,0.01mmol,19.3％)。
化合物26：1H NMR(399MHz,Chloroform-d)δ3.70(s,1H),3.57(d,J＝10.6Hz,1H),2.86(t,J＝13.5Hz,1H),2.37(t,J＝11.3Hz,1H),2.21(d,J＝12.1Hz,1H),2.14–2.02(m,2H),1.97(d,J＝13.1Hz,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)δ212.83,77.31,77.20,76.99,76.67,74.71,73.76,71.92,55.93,55.76,54.82,50.03,48.95,43.78,42.51,38.80,38.60,36.52,36.06,35.89,35.88,35.62,35.55,31.03,29.68,28.40,25.58,24.99,22.29,21.09,18.74,14.16,13.57,13.48,12.04,7.80,1.00.LC-MS:[M+H]+＝433.3。
实施例27
化合物27 3-{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}丙酸的制备。
第一步：将3β-[(3-羟基丙基)氧基]-5α-胆甾-25-醇18(70mg,0.151mmol)溶于二氯甲烷(5mL)中，加入戴斯-马丁氧化剂(64.16mg,0.151mmol)，室温搅拌1小时。TLC(石油醚：乙酸乙酯＝1:1)监测。反应液直接浓缩得到100mg粗品，柱层析纯化(乙酸乙酯：石油醚＝30％)得到3-{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}丙醛27-1(50mg,0.103mmol,68.15％)，白色固体。1H NMR(399MHz,Chloroform-d)δ9.77(t,J＝2.0Hz,1H),3.78(td,J＝6.3,3.1Hz,2H),3.22(dt,J＝11.5,6.2Hz,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).
第二步：将3-{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}丙醛27-1(45mg,0.098mmol)溶于叔丁醇(4mL)和水(0.5mL)中，加入磷酸二氢钠(43.19mg,0.360mmol)，2-甲基-2-丁烯(137.01mg,1.953mmol)，亚氯酸钠(26.50mg,0.293mmol)，室温搅拌1小时。TLC(二氯甲烷：甲醇＝20:1)监测。加水稀释，用乙酸乙酯萃取三次，有机相用盐水洗涤一次，无水硫酸钠干燥，浓缩得到20mg固体，柱层析纯化(甲醇/二氯甲烷＝10％)得到3-{[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}丙酸27(24mg,0.048mmol,48.97％)白色固体。化合物27：1H NMR(400MHz,Methanol-d4)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＝4.0Hz,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) δ174.242,78.790,70.026,63.304,56.451,56.228,54.382,44.646,43.843,42.334,39.970,36.684,36.350,35.704,35.429,35.378,34.873,34.432,31.875,28.548,27.897,27.806,27.780,27.664,23.804,20.894,20.416,17.736,11.265,11.060.LCMS:[Ms-H]-＝475.45.
实施例29
化合物29 3β-[(2-羟基乙基)氧基]-5α-胆甾-4β,25-二醇的制备
第一步：称取乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-甲氧基-6-氧亚基己-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯I-7(77mg,0.17mol,1.0eq.)溶于四氢呋喃(2mL)中,氮气置换后，干冰-丙酮降温至-78℃，加入甲基锂(0.54mL,0.87mmol),随后升至室温搅拌，TLC(石油醚：乙酸乙酯＝10：1)监测,反应结束后，加入6mL水淬灭反应，乙酸乙酯5mL*3萃取，有机相干燥，浓缩，粗品经柱层析(二氯甲烷：甲醇＝300：1～100：1)得胆甾-5(6)-烯-3β,25-二醇29-1(35mg,0.083mmol,47.66％)白色固体。1HNMR(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.
第二步：将胆甾-5(6)-烯-3β,25-二醇29-1(1280mg,3.179mmol,1.0eq.)溶于甲苯(50mL)中，加入叔丁醇钾(3567.00mg,31.789mmol,10.0eq.)，室温搅拌1小时。然后加入溴乙酸叔丁酯(6200.69mg,31.789mmol,10.0eq.)，室温搅拌2小时。TLC(石油醚：乙酸乙酯＝3:1)监测。反应液直接浓缩得到粗品，柱层析纯化(乙酸乙酯：石油醚＝30％)得到{[(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯29-2(950mg,1.746mmol,54.93％)白色固体。1H NMR(399MHz,Chloroform-d)δ5.33(d,J＝5.0Hz,1H),3.98(s,2H),3.27–3.15(m,1H),2.42–2.33(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).
第三步：将{[(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯29-2(950mg,1.746mmol,54.93％)(950mg,1.838mmol,1.0eq.)溶于二氯甲烷(15mL)中，加入吡啶(0.740mL,9.191mmol,5.0eq.)，乙酰氯(0.392mL,5.515mmol,3.0eq.)。40℃搅拌5小时。TLC(石油醚：乙酸乙酯＝5:1)监测。反应液直接浓缩得到粗品，柱层析纯化(乙酸乙酯：石油醚＝20％)得到{[(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-乙酰氧基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯29-3(760mg,1.292mmol,70.28％)白色固体。1H 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,2H),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).
第四步：将{[(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-乙酰氧基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯29-3(760mg,1.360mmol,1.0eq.)溶于氯仿(5mL)中，加入N-甲基吗啉(825.36mg,8.160mmol,6.0eq.)，二氧化硒(198.55mg,1.789mmol,5.0eq.)。75℃搅拌18小时。TLC(石油醚：乙酸乙酯＝5:1)监测。反应液直接浓缩得到粗品，柱层析纯化(乙酸乙酯：石油醚＝40％)得到{[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-1-[(2R)-6-乙酰氧基-6-甲基庚-2-基]-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯29-4(320mg,0.529mmol,38.89％)白色固体。1H NMR(399MHz,Chloroform-d)δ5.68(d,J＝4.5Hz,1H),4.19–4.11(m,2H),3.93(d,J＝16.8Hz,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＝8.7Hz,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).
第五步：将{[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-1-[(2R)-6-乙酰氧基-6-甲基庚-2-基]-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯29-4(320mg,0.557mmol,1.0eq.)溶于乙酸乙酯(10mL)，乙酸(2mL)中，加入二氧化铂(160mg,0.705mmol,1.3eq.)。氢气环境下40℃搅拌15min。TLC(石油醚：乙酸乙酯＝5:1)监测。反应液过滤，滤液浓缩得到粗品，柱层析纯化(乙酸乙酯：石油醚＝30％)得到{[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-1-[(2R)-6-乙酰氧基-6-甲基庚-2-基]-6-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯29-5(200mg,0.329mmol,59.17％)白色固体。1H 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),1.16–1.06(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).
第六步：将{[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-1-[(2R)-6-乙酰氧基-6-甲基庚-2-基]-6-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-基]氧基}乙酸-2-甲基丙-2-基酯29-5(40mg,0.069mmol，1.0eq.)溶于四氢呋喃(5mL)中，加入四氢铝锂(13.16mg,0.347mmol,5.0eq.)。室温搅拌1h。TLC(石油醚：乙酸乙酯＝5:1)监测。反应液加水淬灭，过滤，滤液浓缩得到粗品，柱层析纯化(乙酸乙酯：石油醚＝50％)得到3β-[(2-羟基乙基)氧基]-5α-胆甾-4β,25-二醇29(20mg,0.041mmol,58.96％)白色固体。化合物29：1H NMR(399MHz,Chloroform-d)δ3.88(s,1H),3.73(q,J＝3.8,3.3Hz,2H),3.62(t,J＝4.6Hz,2H),3.25(dt,J＝11.9,4.3Hz,1H),1.94(d,J＝12.3Hz,1H),1.83–1.73(m,5H),1.70(s,6H),1.54(d,J＝9.1Hz,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-H2O]+＝429.40；CAD:92.03％.
实施例31
化合物31 25-羟基-3β-[(2-羟基乙基)氧基]-5α-胆甾-4-酮的制备。
第一步：将3β-[(2-羟基乙基)氧基]-5α-胆甾-4β,25-二醇29(86mg,0.185mmol,1.0eq.)溶于二氯甲烷(5mL)中，加入咪唑(62.99mg,0.925mmol,5.0eq.)，叔丁基二甲基氯硅烷(83.67mg,0.555mmol,3.0eq.)。室温搅拌2h。TLC(石油醚：乙酸乙酯＝5:1)监测。反应液浓缩得到粗品，柱层析纯化(乙酸乙酯：石油醚＝20％)得到3β-[(4,4,5,5-四甲基-3-氧杂-4-硅杂己-1-基)氧基]-5α-胆甾-4β,25-二醇31-1(65mg,0.107mmol,57.63％)白色固体。1H 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).
第八步：将3β-[(4,4,5,5-四甲基-3-氧杂-4-硅杂己-1-基)氧基]-5α-胆甾-4β,25-二醇31-1(60mg,0.104mmol,1.0eq.)溶于二氯甲烷(1mL)中，加入戴斯-马丁氧化剂(43.95mg,0.104mmol,1.0eq.)。室温搅拌1h。TLC(石油醚：乙酸乙酯＝5:1)监测。反应液浓缩得到粗品，柱层析纯化(乙酸乙酯：石 油醚＝20％)得到25-羟基-3β-[(4,4,5,5-四甲基-3-氧杂-4-硅杂己-1-基)氧基]-5α-胆甾-4-酮31-2(50mg,0.082mmol,79.44％)白色固体。1H NMR(399MHz,Chloroform-d)δ4.01(dd,J＝12.1,7.1Hz,1H),3.83–3.73(m,3H),3.44(dt,J＝11.3,5.2Hz,1H),2.25(s,1H),2.02(dd,J＝38.0,12.2Hz,2H),1.83(dd,J＝37.0,11.2Hz,3H),1.72(d,J＝13.6Hz,3H),1.50(d,J＝20.7Hz,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).
第九步：将25-羟基-3β-[(4,4,5,5-四甲基-3-氧杂-4-硅杂己-1-基)氧基]-5α-胆甾-4-酮31-2(40mg,0.069mmol,1.0eq.)溶于四氢呋喃(2mL)中，加入四丁基氟化铵(0.5mL,0.500mmol,7.2eq.)。室温搅拌1h。TLC(石油醚：乙酸乙酯＝3:1)监测。反应液加水，用乙酸乙酯萃取三次，有机相干燥后浓缩得到粗品，柱层析纯化(甲醇/乙酸乙酯＝5％)得到25-羟基-3β-[(2-羟基乙基)氧基]-5α-胆甾-4-酮31(12mg,0.025mmol,35.54％)，白色固体。
化合物31：1H NMR(400MHz,Methanol-d4)δ4.08(dd,J＝11.8,7.2Hz,1H),3.70–3.59(m,3H),3.52(dd,J＝9.3,3.4Hz,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＝19.4,6.4Hz,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＝13.2Hz,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％.
实施例33
化合物33[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]乙腈的制备
第一步：室温下50mL圆底烧瓶中，将4-(羟基甲基)-5α-胆甾-3β,25-二醇13-1(120mg,0.28mmol)溶解于1,2-二氯乙烷(5mL)，在室温下加入三乙胺(170mg，1.68mmol),4-二甲氨基吡啶(34mg,0.28 mmol)和对甲基苯磺酰氯(107mg，0.56mmol)，随后在50℃下搅拌8小时。TLC(石油醚:乙酸乙酯＝2:1)监测反应，反应结束后，用水(30mL)淬灭，乙酸乙酯(20mL×3)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝2：1)，得到白色固体4-甲基苯磺酸-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]甲基酯33-1(60mg，纯度:90％，产率：36.36％)。1HNMR(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,22H),1.21(s,6H),1.13–0.93(m,6H),0.90(d,J＝6.4,3H),0.61(s,6H).
第二步：室温下50mL圆底烧瓶中，将4-甲基苯磺酸-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]甲基酯33-1(40mg,0.068mmol)溶解于N,N-二甲基甲酰胺(1mL)，在室温下加入氰化钠(20mg，0.41mmol)，随后在100℃下搅拌16小时，TLC(石油醚:乙酸乙酯＝3:1)监测反应，反应结束后，用水(20mL)淬灭，乙酸乙酯(10mL×3)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝3：1)，得到白色固体[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]乙腈33(15.71mg，纯度:77.49％，产率：40.39％)。
化合物33：1H NMR(400MHz,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).13C 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-2H2O]+＝408.3
实施例34
化合物34 24,24-二氟-5α-胆甾-3β,4β,25-三醇的制备。
第一步将(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟己酸乙酯24-3(300mg,0.606mmol,1eq)溶于氯仿(10mL)，向溶液中加入二氧化硒(201.8mg,1.82mmol,3eq)，N-甲基吗啉(153.2mg,1.52mmol,2.5eq)，室温搅拌20小时。反应完成后，加入30mL水，用乙酸乙酯(30mL*3)萃取，将得到的有机相用无水硫酸钠干燥后旋干得到粗品，粗品经硅胶过柱(石油醚:乙酸乙酯＝10:1)纯化得到(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟己酸乙酯34-1(155mg,0.304mmol,收率：50％)。1H NMR(400MHz,CDCl3)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,3H),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).
第二步(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟己酸乙酯34-1(155mg,0.304mmol,1eq)溶于乙酸乙酯(15mL)中,然后向其中加入二氧化铂(50％)(75mg)，室温搅拌12小时,反应完成后,过滤,然后旋干有机溶剂得到产物(5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟己酸乙酯34-2(70mg,0.137mmol,收率：46％)为油状化合物。1H NMR(400MHz,cdcl3)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,3H),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).
第三步将化合物(5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]-2,2-二氟己酸乙酯34-2(70mg,0.137mmol,1eq)溶于四氢呋喃(5mL)中，置换氮气后缓慢滴加甲基溴化镁(2.4M，0.6mL，10eq)，室温搅拌3小时,TLC(石油醚:乙酸乙酯＝1:1)监测,反应完成后向其中加入水摧灭,然后用乙酸乙酯(50mL)萃取,有机相用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,过滤,旋干得到粗品,然后经过硅胶分离纯化(石油醚:乙酸乙酯＝1:1)得到(1R,3aS,3bR,7S,9aS,9bS,11aR)-4,4-二氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-醇34(11mg,0.024mmol,收率：18％)。
化合物34：1H NMR(400MHz,CDCl3)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＝6.1Hz,2H),0.85(d,J＝6.6Hz,3H),0.81(s,1H),0.59(s,3H),0.56–0.49(m,1H).13C NMR(101MHz,CDCl3)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。
实施例41&65
化合物41(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4,4-二氟-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]十六氢-1H-环戊并[1,2-a]菲-6,7-二醇
化合物65(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-4-氟-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6,7-二醇的制备。
第一步：将(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-二氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1’,2’:1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己醛I和(5R)-5-[(3aS,5aR,5bR,7aR,8R,10aR,12bR)-11-氟-2,2,5a,7a-四甲基4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-十四氢-3aH-环戊并[1’,2’:7,8]菲并[1,2-d][1,3]二氧杂环戊熳-8-基]己醛II的混合物(70mg,0.150mmol,1.0eq)溶解于四氢呋喃(5mL)中，完全溶解后依次向反应体系中加入氟化铯(11.39mg,0.075mmol，0.5eq)，(三氟甲基)三甲基硅烷(106.7mg,0.750mmol,5eq)，加料完毕后在氮气置换三次，室温下搅拌1小时。TLC(石油醚:乙酸乙酯＝5:1)监测反应完全后，加入四丁基氟化铵(1mL,1mmol，1mol/L),加料完毕后在氮气置换三次，室温下搅拌1小时。TLC(石油醚:乙酸乙酯＝5:1)监测反应完全后，用水(10mL)萃灭，反应液用水(～10mL)洗涤三次，无水硫酸钠干燥，浓缩，得到粗品。粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝95:5to 85：15)纯化，得到白色固体(6R)-6-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-二氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1’,2’:1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]-1,1,1-三氟庚-2-醇41-1和(6R)-1,1,1-三氟-6-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-十四氢-3aH-环戊并[1’,2’:7,8]菲并[1,2-d][1,3]二氧杂环戊熳-8-基]庚-2-醇65-1混合物(50mg，0.089mmol，纯度80.0％，以41-1计算收率59.62％)。
化合物41-1：1HNMR(400MHz,CDCl3)δ3.94(dt,J＝8.3,6.2Hz,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＝3.4Hz,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).
第二步：将原料(6R)-6-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11,11-二氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1’,2’:1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]-1,1,1-三氟庚-2-醇41-1和(6R)-1,1,1-三氟-6-[(3aS,5aR,5bR,7aR,8R,10aR,12aR,12bR)-11-氟-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,12,12a,12b-十四氢-3aH-环戊并[1’,2’:7,8]菲并[1,2-d][1,3]二氧杂环戊熳-8-基]庚-2-醇65-1混合物(50mg,0.093mmol,1.0eq)溶于四氢呋喃(5mL)中,缓慢滴加盐酸(1mL,3mol/L),氮气置换三次,室温搅拌1hr。TLC(石油醚:乙酸乙酯＝1：1)监测反应。反应液使用饱和碳酸氢钠溶液(3×5mL)溶液洗涤。合并有机相用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，浓缩得到粗产物。粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝90：10to80：20)纯化，(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4,4-二氟-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]十六氢-1H-环戊并[1,2-a]菲-6,7-二醇41和(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-4-氟-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6,7-二醇的混合物65混合物(40mg,基于41计算收率69.16％),直接投下一步。
第三步：化合物(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4,4-二氟-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]十六氢-1H-环戊并[1,2-a]菲-6,7-二醇41和(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-4-氟-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6,7-二醇的混合物65混合物(23mg,0.046mmol,1.0eq)加入到盛有二氯甲烷(2mL)的反应瓶中,加入三乙胺(0.045mL,0.324mmol,7.0eq)和苯甲酰氯(0.027mL,0.232mmol,5.0eq)后,在室温(20℃)下搅拌18hr；TLC(二氯甲烷:甲醇＝20:1)监测反应。反应液加入水(20mL)，用乙酸乙酯(20mL*2)萃取，合并有机相，经水(20mL*2)洗涤，无水硫酸钠干燥，过滤，滤液减压旋干(水泵，45℃)，得到粗品，经prep-TLC(二氯甲烷:甲醇＝20:1)分离纯化和SFC分离(仪器：Waters Acquity UPCC；色谱柱：Daicel CHIRALPAK ID_3,3.0*150mm,3um；流动相：A/B：CO2/MeOH(0.1％DEA)＝75/25；流速：2.0毫升/分钟；柱温：37度)得到产物41-2苯甲酸-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4,4-二氟-6-羟基-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]十六氢-1H-环戊并[1,2-i]菲-7-基酯(20mg,0.030mmol,64.70％,保留时间tR＝1.096min)为白色固体。产物65-2：苯甲酸-(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-4-氟-6-羟基-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯(10mg,0.010mmol,22.38％,保留时间tR＝1.280min)为白色固体。
化合物41-2：1H NMR(400MHz,CDCl3)δ8.08(d,J＝7.6Hz,2H),7.62(t,J＝7.4Hz,1H),7.48(t,J＝7.7Hz,2H),5.61–5.48(m,1H),3.73(s,1H),3.59(d,J＝10.7Hz,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).19F NMR(377MHz,CDCl3)δ-76.92,-77.02,-88.65,-89.28,-110.63,-111.26.
化合物65-2：19F NMR(377MHz,CDCl3)δ-76.91,-77.04,-109.74.
第四步：化合物苯甲酸-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4,4-二氟-6-羟基-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]十六氢-1H-环戊并[1,2-i]菲-7-基酯41-2(20mg,0.033mmol,1.0eq)加入到盛有甲醇(0.5mL)和四氢呋喃(1mL)的反应瓶中，加入氢氧化锂(13.97mg,0.333mmol)的水(0.5mL)溶液，升温到40℃搅拌2hr；TLC(石油醚:乙酸乙酯＝1:1)监测反应显示原料消失,有极性变大的主点生成；反应液加入水(20mL)，用乙酸乙酯(20mL*3)萃取，合并有机相，经水(20mL*2)洗涤，无水硫酸钠干燥，过滤，滤液减压旋干(水泵，45℃)，得到粗品，经柱层析(石油醚:乙酸乙酯＝1:0-2:1)分离纯化，得到产物(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4,4-二氟-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]十六氢-1H-环戊并[1,2-a]菲-6,7-二醇41(16.40mg,0.033mmol,99.21％)为白色固体。
化合物41：1H NMR(400MHz,CDCl3)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).19F NMR(377MHz,CDCl3)δ-80.03,-88.64,-89.27,-110.64,-111.26.19F NMR(377MHz,CDCl3)δ-76.92,-77.02,-88.65,-89.28,-110.63,-111.26.
化合物苯甲酸-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4-氟-6-羟基-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]十六氢-1H-环戊并[1,2-i]菲-7-基酯65-2(10mg,0.017mmol,1.0eq)加入到盛有甲醇(0.5mL)和四氢呋喃(1mL)的反应瓶中，加入氢氧化锂(7.20mg,0.172mmol)的水(0.5mL)溶液，升温到40℃搅拌2hr；TLC(石油醚:乙酸乙酯＝1:1)监测反应；反应液加入水(20mL)，用乙酸乙酯(20mL*3)萃取，合并有机相，经水(20mL*2)洗涤，无水硫酸钠干燥，过滤，滤液减压旋干(水泵，45℃)，得到粗品，经柱层析(石油醚:乙酸乙酯＝1:0-2:1)分离纯化，得到产物(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-4-氟-9a,11a-二甲基-1-[(2R)-7,7,7-三氟-6-羟基庚-2-基]-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6,7-二醇65(1.89mg,0.004mmol,21.44％)为白色固体。
化合物65：1H NMR(400MHz,CDCl3)δ3.91(s,1H),3.85(s,1H),3.59(d,J＝11.0Hz,1H),2.66(t,J＝12.5Hz,1H),2.07–1.98(m,2H),1.94(d,J＝16.5Hz,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＝6.5Hz,3H),0.65(s,3H).19F NMR(376MHz,CDCl3)δ-80.03,-109.71.19F NMR(377MHz,CDCl3)δ-76.91,-77.04,-109.74.
实施例53
化合物53 4-(1-羟基乙基)-5α-胆甾-3β,25-二醇的制备

第一步室温下在50mL的圆底烧瓶中，将3β-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-25-羟基-5α-胆甾-4-甲醛23-3(23mg,0.042mmol)溶解于无水四氢呋喃(1.5mL)，反应氮气保护下，降温到0℃，滴加三摩尔每升的甲基溴化镁四氢呋喃溶液(0.07mL，0.021mmol)，随后转至室温搅拌2小时，TLC(石油醚:乙酸乙酯＝4：1)监测反应.反应结束后，体系降温至0℃，用饱和氯化铵水溶液(20mL)淬灭，乙酸乙酯(10mL×3)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，得到白色固体3β-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-4-(1-羟基乙基)-5α-胆甾-25-醇53-1(17mg，纯度:90％，产率：64.64％)。1HNMR(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).
第二步室温下在50mL的圆底烧瓶中，将3β-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-4-(1-羟基乙基)-5α-胆甾-25-醇53-1(17mg,0.03mmol)溶解于一摩尔每升的四丁基氟化铵四氢呋喃溶液(0.5mL)，在室温下搅拌2小时。TLC(石油醚:乙酸乙酯＝1：1)监测反应，反应结束后，体系降温至0℃，用饱和氯化铵水溶液(10mL)淬灭，乙酸乙酯(5mL×3)萃取，有机相饱和盐水(10mL)洗涤，无水硫酸钠干燥，硅胶柱层析纯化(石油醚:乙酸乙酯＝1：1)，得到白色固体4-(1-羟基乙基)-5α-胆甾-3β,25-二醇53(5.09mg，纯度:95.12％，产率：35.73％)。
化合物53：1H NMR(400MHz,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).13C NMR(101MHz,CDCl3)δ＝104.61,103.05,101.06,71.13,69.23,56.48,56.37,56.18,44.43,42.48,39.95,38.55,36.44,35.88,35.75,35.73,33.64,33.59,31.45,30.97,30.36,29.37,29.21,29.04,28.26,26.29,25.61,24.80,24.27,21.07,20.99,20.92,20.78,18.64,14.96,12.01.LC-MS：[M+H-2H2O]+＝413.55
实施例55
化合物55 4-溴-5α-胆甾-3β,25-二醇的制备
第一步在100mL圆底烧瓶中，将(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-8(500mg,1.09mmol)溶解于乙酸乙酯(30mL)，在室温下加入10％钯碳(250mg)和乙酸(0.3mL)，在室温下将混合物在氢气氛围中搅拌5小时。TLC(乙酸乙酯/石油醚＝8：1)监测反应。反应结束后，用乙酸乙酯(60mL)稀释反应液，通过硅藻土过滤，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝8：1)，得白色固体(5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯55-1(390mg，纯度：90％，产率：62.72％)。1H 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).
第二步室温下50mL圆底烧瓶中，将(5R)-5-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯55-1(250mg,0.54mmol)溶解于1,2-二氯乙烷(10mL)，在0℃下加入三苯基膦(142mg，0.54mmol)的1,2-二氯乙烷(0.6mL)溶液，反应液在室温搅拌10分钟，在0℃下向反应液中加入四溴化碳(180mg，0.54mmol)的1,2-二氯乙烷(0.9mL)溶液，随后将反应液升到室温，在50℃下搅拌16小时。TLC(石油醚:乙酸乙酯＝5:1)监测反应。反应结束后，用水(30mL)淬灭，乙酸乙酯(20mL×3)萃取，有机相饱和盐水(40mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝5：1)，得到白色固体(5R)-5-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-乙酰氧基-6-溴-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯55-2(150mg，纯度:90％，产率：47.57％)。1H 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).
第三步：将反应物(5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-溴-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯55-2(150mg,0.337mmol,1eq)溶于四氢呋喃(10mL)中，并将反应体系置换成氮气氛围。将反应体系冷却至0℃后，缓慢加入甲基氯化镁(1.124mL,3.373mmol,1mol/L)，并保持在室温搅拌1h。TLC(石油醚:乙酸乙酯＝5：1)监测反应。将20mL水加入到反应体系中，使用乙酸乙酯(25mL×3)萃取，收集有机相，使用无水硫酸钠干燥，有机相真空旋干得到粗品。粗品溶于乙酸乙酯硅胶柱层析纯化(石油醚:乙酸乙酯＝88:12)得到4-溴-5α-胆甾-3β,25-二醇55(16mg,纯度：90.51％,收率：10.66％)。
化合物55：1H 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).
实施例56
化合物56 4β-[(乙氧基甲基)氧基]-3-甲基-5α-胆甾-3,25-二醇的制备。

第一步：将乙酸-(1R,7S,9aR,11aR)-6-羟基-1-[(2R)-6-甲氧基-6-氧亚基己-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯55-1(100mg,0.216mmol)溶解在中N,N-二异丙基乙胺(5mL),加入[(氯甲基)氧基]乙烷(204.21mg,2.16mmol)，130℃反应1.5h，TLC(石油醚：乙酸乙酯＝5:1)监测和核磁监测，原料无剩余，用乙酸乙酯和水洗萃后用无水硫酸钠干燥浓缩，柱层析(石油醚：乙酸乙酯＝100：1～20:1)纯化得乙酸-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-[(乙氧基甲基)氧基]-1-[(2R)-6-甲氧基-6-氧亚基己-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯56-1(60mg,0.106mmol,49.04％)白色固体。1HNMR(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).
第二步：将乙酸-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-[(乙氧基甲基)氧基]-1-[(2R)-6-甲氧基-6-氧亚基己-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯56-1(60mg,0.115mmol)溶解在甲醇(5mL)，室温加入碳酸钾(158.70mg,1.150mmol)，室温搅拌5h，TLC(石油醚：乙酸乙酯＝5:1)监测；反应结束后加入水和乙酸乙酯萃取，有机相分离干燥粗品，经柱层析(石油醚：乙酸乙酯＝60:1～3：1)纯化得(5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-[(乙氧基甲基)氧基]-7-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯56-2(40mg,0.079mmol,58.89％)白色固体。1H 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).
第三步：将(5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-[(乙氧基甲基)氧基]-7-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯56-2(20mg,0.042mmol)溶解在二氯甲烷(5mL)，室温加入N-甲基吗啉氧化物(7.34mg,0.063mmol)和四丙基高钌酸铵(1.47mg,0.004mmol)，室温搅拌1h，TLC(石油醚：乙酸乙酯＝5:1)监测；反应结束后用硅藻土过滤，滤液浓缩，经柱层析(石油醚：乙酸乙酯＝60:1～3：1)纯化得(5R)-5-[(1R,3aS,3bS,5aR,6R,9aR,9bS,11aR)-6-[(乙氧基甲基)氧基]-9a,11a-二甲基-7-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯56-3(15mg,0.029mmol,69.08％)白色固体。1H 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)。
第四步：将(5R)-5-[(1R,3aS,3bS,5aR,6R,9aR,9bS,11aR)-6-[(乙氧基甲基)氧基]-9a,11a-二甲基-7-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯56-3(20mg,0.042mmol)溶解在四氢呋喃(3mL)，零度氮气保护下滴加甲基锂(9.22mg,0.42mmol)后，室温搅拌2h，TLC(石油醚：乙酸乙酯＝2:1)监测；反应结束后加入水和乙酸乙酯萃取，有机相分离干燥粗品，经柱层析(石油醚：乙酸乙酯＝50:1～1：1)纯化得4β-[(乙氧基甲基)氧基]-3-甲基-5α-胆甾-3,25-二醇56(10mg,0.019mmol,45.90％)白色固体。
化合物56：1HNMR(400MHz,Chloroform-d)δ4.78(d,J＝6.6Hz,1H),4.60(d,J＝6.6Hz,1H),3.73–3.57(m,2H),3.15(s,1H),1.94(d,J＝12.6Hz,1H),1.74(t,J＝14.7Hz,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).13CNMR(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+NH4]+＝491.42；[Ms-H2O-OH]+＝493.42.HPLC:94.97％(CAD).
实施例59
化合物59 3-甲基-5α-胆甾-4β,3,25-三醇的制备。
将4β-[(乙氧基甲基)氧基]-3-甲基-5α-胆甾-3,25-二醇56(50mg,0.101mmol)溶解在四氢呋喃(3mL)，室温加入对甲苯磺酸(174.08mg,1.014mmol)，加热40℃搅拌1h，TLC(石油醚：乙酸乙酯＝1:1)监测；反应结束后加入水和乙酸乙酯萃取，有机相分离，无水硫酸钠干燥粗品，经柱层析(石油醚：乙酸乙酯＝50:1～1：1)纯化得3-甲基-5α-胆甾-4β,3,25-三醇59(20mg,0.044mmol,45.35％)白色固体。
化合物59：1H 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,3H),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+NH4]+＝433.38；[Ms-H2O-OH]+＝435.38.HPLC:93.78％(CAD).
实施例60
化合物60 24-氟-5α-胆甾-3β,4β,25-三醇的制备。

第一步：称取乙酸-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-羟基-1-[(2R)-6-甲氧基-6-氧亚基己-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯IV-1(300mg,0.65mol,1.0eq.)溶于无水甲醇(5mL)中,加入碳酸钾(897mg,6.5mmol),接着升至25℃搅拌16小时，TLC(石油醚：乙酸乙酯＝2:1)监测,反应结束后。将反应液过滤后浓缩，所得粗品经柱层析(石油醚：乙酸乙酯＝0～50％)纯化得(5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-二羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯IV-2(210mg,0.5mmol,76.9％)白色固体。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).
第二步：称取(5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-二羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯IV-2(250mg,0.59mmol)溶于丙酮(20mL)中，加入4A分子筛(250mg)，对甲苯磺酸(203mg,1.18mmol)后室温搅拌，TLC(石油醚：乙酸乙酯＝2:1)监测。反应结束后，过滤除去不溶物，滤液浓缩后经柱层析(石油醚：乙酸乙酯＝0～30％)纯化后得(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己酸甲酯IV-3(220mg,0.48mmol,81.3％)白色固体。1H 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,2H),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).
第三步在三口瓶中将(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1’,2’:1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己酸甲酯IV-3(370mg,0.803mmol,1.0eq.)溶于四氢呋喃(7mL)，加入三甲基氯化硅(0.440mL,2.409mmol,3.0eq.)。将三口瓶内置换至氮气环境，然后降温至-78℃，加入二异丙基氨基锂(172.08mg,1.606mmol,2.0eq.)。反应液随后升温至0℃搅拌15分钟。加入N-氟代双苯磺酰胺(506.50mg,1.606mmol,2.0eq.)的四氢呋喃(1mL)溶液。反应液升温至室温搅拌1h。TLC(石油醚：乙酸乙酯＝10:1)监测。待原料完全反应完，向反应液中加水淬灭。得到的混合液用乙酸乙酯萃取三次，合并的有机相用盐水洗涤一次，无水硫酸钠干燥，浓缩得到粗品，柱层析纯化(乙酸乙酯/石油醚＝0–20％)得到(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1’,2’:1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]-2-氟己酸甲酯白色固体60-1(200mg,0.397mmol,49.42％)。1H 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),1.37(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＝8.7,4.3Hz,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；
第四步将(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1’,2’:1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]-2-氟己酸甲酯60-1(100mg,0.209mmol,1eq.)溶于四氢呋喃(5mL)中。在氮气环境下降温至0℃，加入甲基锂(0.653mL,1.045mmol,5.0eq.)。升温至室温搅拌1小时。TLC(石油醚：乙酸乙酯＝10:1)监测。反应完全后，加入少量水淬灭，混合液直接浓缩得到粗品，柱层析纯化(乙酸乙酯/石油醚＝0–30％)得到(6R)-6-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1’,2’:1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]-3-氟-2-甲基庚-2-醇白色固体60-2(75mg,0.149mmol,71.24％)。1H 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),1.02(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).
第五步将(6R)-6-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1’,2’:1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]-3-氟-2-甲基庚-2-醇60-2(75mg,0.157mmol)溶于四氢呋喃(5mL)中，加入盐酸(2mL,4.000mmol,25.5eq.)。室温搅拌1小时。TLC(石油醚：乙酸乙酯＝5:1)监测。反应液加水稀释，用乙酸乙酯萃取三次，有 机相合并，用盐水洗涤一次，无水硫酸钠干燥。浓缩得到粗品，柱层析纯化(乙酸乙酯/石油醚＝0-60％)得到24-氟-5α-胆甾-3β,4β,25-三醇白色固体60(40mg,0.087mmol,55.29％)。
化合物60：1H 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),1.37(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＝8.7,4.3Hz,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+Na]+＝461.45。
实施例61
化合物61,3,25-二羟基-3-甲基-5α-胆甾-4-酮的制备。
将3-甲基-5α-胆甾-4β,3,25-三醇59(20mg,0.046mmol)溶解二氯甲烷(3mL)，室温加入N-甲基吗啉氧化物(8.08mg,0.069mmol)和四丙基高钌酸铵(1.61mg,0.005mmol)，室温搅拌1h，TLC(石油醚：乙酸乙酯＝2:1)监测；反应结束后用硅藻土过滤，滤液浓缩，经柱层析(石油醚：乙酸乙酯＝50:1～1：1)纯化得3,25-二羟基-3-甲基-5α-胆甾-4-酮61(10mg,0.020mmol,85.85％)白色固体。
化合物61：1HNMR(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+NH4]+＝433.38；[M-H2O-OH]+＝435.38.
实施例66
化合物66：(1R,3aR,7S,9aS,9bR,11aR)-4-氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-7-醇的制备。
第一步：将(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-7(1g,2.25mmol,1eq)溶于四氢呋喃(10mL)，向溶液中加入乙酸钴(80mg,0.45mmol,0.2eq)，N-羟基临苯二甲酰亚胺(0.15g,0.90mmol,0.4eq)，过氧化叔丁醇(1.01g,11.25mmol,5eq)，室温搅拌20小时。反应完成后，加入30mL水，用乙酸乙酯(30mL*3)萃取，将得到的有机相用无水硫酸钠干燥后旋干得到粗品，粗品经硅胶过柱(石油醚/乙酸乙酯＝10/1)纯化得到(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-4-氧亚基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯66-1(0.6g,1.31mmol,收率：58％).1H NMR(400MHz,CDCl3)δ5.70(d,J＝1.5Hz,1H),4.80–4.64(m,1H),3.67(s,3H),2.48(dddd,J＝15.6,12.1,9.1,2.4Hz,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).
第二步：将化合物(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-4-氧亚基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯66-1(0.6g,1.31mmol,1eq)溶于乙酸乙酯(15mL)中,然后向其中加入钯碳(10％)(120mg)，室温搅拌12小时,反应完成后,过滤,然后旋干有机溶剂得到产物(5R)-5-[(1R,3aS,3bR,7S,9aS,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-4-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯66-2(450mg,0.977mmol,收率：75％)为油状化合物。1H NMR(400MHz,cdcl3)δ5.36–5.24(m,1H),4.30(s,3H),3.00–2.81(m,5H),2.70–2.58(m,5H),2.55–2.47(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).
第三步：将化合物(5R)-5-[(1R,3aS,3bR,7S,9aS,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-4-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯66-2(150mg,0.326mmol,1.0eq),DAST(157mg,0.977mmol,3eq)溶于四氢呋喃(5mL)中，80℃搅拌3小时,TLC检测,反应完成后向其中加入水摧灭,然后用乙酸乙酯(50mL)萃取,有机相用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,过滤,旋干得到粗品,然后经过硅胶分离纯化(石油醚:乙酸乙酯＝1:1)得到(R)-5-((3S，5R，9R，10S，13R，14R，17R)-3-乙酰氧基-7-氟-10,13-二甲基-2,3,4,5,6,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊并[a]菲17-基)己酸甲酯66-3和(R)-5-((3S，5R，8R，9S，10S，13R，14S，17R)-3-乙酰氧基-7,7-二氟-10,13-二甲基十六氢-1H-环戊基[a]菲17-基)己酸甲酯66-3’的混合物(70mg,0.145mmol,以66-3’计算收率：45％)。
化合物66-3’:1H NMR(400MHz,CDCl3)δ4.75–4.63(m,1H),3.66(s,3H),2.34–2.22(m,2H),2.10–1.91(m,5H),1.84(ddd,J＝9.3,7.6,3.7Hz,3H),1.78–1.65(m,5H),1.57–1.49(m,3H),1.45–1.22(m,7H),1.09(ddd,J＝28.8,14.4,9.4Hz,5H),0.94(t,J＝5.4Hz,3H),0.85(d,J＝11.0Hz,3H),0.71–0.62(m,3H).
第四步：将(R)-5-((3S，5R，9R，10S，13R，14R，17R)-3-乙酰氧基-7-氟-10,13-二甲基-2,3,4,5,6,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊并[a]菲17-基)己酸甲酯66-3和(R)-5-((3S，5R，8R，9S，10S，13R，14S，17R)-3-乙酰氧基-7,7-二氟-10,13-二甲基十六氢-1H-环戊基[a]菲17-基)己酸甲酯66-3’的混合物(70mg,0.145mmol,1eq)溶于四氢呋喃(5mL)中，置换氮气后缓慢滴加甲基溴化镁(2.4M)(0.6mL，10eq)，室温搅拌3小时,TLC检测,反应完成后向其中加入水摧灭,然后用乙酸乙酯(50mL)萃取,有机相用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,过滤,旋干得到粗品,然后经过硅胶分离纯化(石油醚:乙酸乙酯＝1:1)得到(3S，5R，9R，10S，13R，14R，17R)-7-氟-17-((R)-6-羟基-6-甲基庚烷-2-基)-10,13-二甲基-2,3,4,5,6,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊二烯[a]菲-3-醇66-4和(3S，5R，8R，9S，10S，13R，14S，17R)-7,7-二氟-17-((R)-6-羟基-6-甲基庚烷-2-基)-10,13-二甲基六氢-1H-环戊并[a]菲-3-醇66-4‘混合物(22mg,0.05mmol,以66-4’计算收率：34％),直接投下一步。
第五步：将化合物(3S，5R，9R，10S，13R，14R，17R)-7-氟-17-((R)-6-羟基-6-甲基庚烷-2-基)-10,13-二甲基-2,3,4,5,6,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊二烯[a]菲-3-醇66-4和(3S，5R，8R，9S，10S，13R，14S，17R)-7,7-二氟-17-((R)-6-羟基-6-甲基庚烷-2-基)-10,13-二甲基六氢-1H-环戊并[a]菲-3-醇66-4‘混合物(1.3g,2.95mmol,1eq)溶于二氯甲烷(25mL)中，缓慢滴加三乙胺(1.19g，11.80mmol，4eq)，加入DMAP(0.29g，2.36mmol，0.8eq)后，室温下缓慢滴加苯甲酰氯(1.04g，7.38mmol，2.5eq)，室温搅拌1小时,TLC检测,反应完成后向其中加入水摧灭,然后用乙酸乙酯(50mL)萃取,有机相用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,过滤,旋干得到粗品,然后经过硅胶分离纯化(石油醚:乙酸乙酯＝1:1)得到66-5和66-5’混合物(1.5g,2.82mmol,收率：95％)。经过SFC拆分(仪器：Waters Acquity UPCC；柱：Daicel CHIRALPAK OZ3，3*150mm，3um；流动相：A/B:CO2/MeOH(0.1％DEA)＝70/30；流速：2.0毫升/分钟；柱温度：37度)得到(6R)-6-[(1R,3aS,3bR,7S,9aS,9bS,11aR)-7-(苄基氧基)-4,4-二氟-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-醇66-5’(1g，保留时间tR＝1.178min)和(1R,3aR,7S,9aS,9bR,11aR)-4-氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-7-醇66-5(200mg，保留时间tR＝1.601min)。
化合物66-5：1HNMR(400MHz,CDCl3)δ8.07–7.99(m,2H),7.55(t,J＝7.4Hz,1H),7.43(t,J＝7.6Hz,2H),4.96(d,J＝5.0Hz,1H),1.22(s,6H),0.96–0.85(m,6H),0.72–0.64(m,3H).19F NMR(377MHz,CDCl3)δ-110.61.
化合物66-5‘：19F NMR(377MHz,CDCl3)δ-88.95,-89.58,-110.84,-111.47.
第六步：将化合物(1R,3aR,7S,9aS,9bR,11aR)-4-氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-7-醇66-5(100mg,0.2mmol,1eq)溶于四氢呋喃(4mL)和甲醇(2ml)中，加入氢氧化锂一水合物(82mg,2mmol,10eq)，40度下搅拌3小时,TLC检测,反应完成后向其中加入水摧灭,然后用乙酸乙酯(50mL)萃取,有机相用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,过滤,旋干得到粗品,然后经过硅胶分离纯化(石油醚:乙酸乙酯＝1:1)得到(1R,3aR,7S,9aS,9bR,11aR)-4-氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-7-醇66(70mg,0.16mmol,收率：85％)。
化合物66：1H NMR(400MHz,CDCl3)δ3.60(t,J＝4.4Hz,1H),2.08–1.90(m,3H),1.83(dddd,J＝19.8,15.3,9.9,5.8Hz,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＝6.5Hz,3H),0.82(s,3H),0.64(s,3H).
实施例79
化合物79(1R,3aR,5aR,7S,9aR,9bR,11aR)-4-氟-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6-酮的制备。
第一步室温下在100mL的圆底烧瓶中将(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-8(1g,2.17mmol)溶解于无水四氢呋喃(20mL)，反应氮气保护下，降温到0℃，滴加甲基溴化镁(7.23mL， 21.70mmol)，随后转至室温搅拌2小时，TLC(石油醚:乙酸乙酯＝2：1)监测反应，反应结束后，体系降温至0℃，用饱和氯化铵水溶液(60mL)淬灭，乙酸乙酯(50mL×3)萃取，有机相饱和盐水(80mL)洗涤，无水硫酸钠干燥，硅胶柱层析纯化(石油醚:乙酸乙酯＝2：1)，得到白色固体胆甾-6(5)-烯-3β,4β,25-三醇79-1(800mg，纯度:90％，产率：79.38％)。1H NMR(400MHz,CDCl3)δ5.68(d,J＝3.3Hz,1H),4.14(s,1H),3.56(d,J＝11.1Hz,1H),2.22–1.72(m,7H),1.72–1.31(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).
第二步室温下在100mL的圆底烧瓶中，将胆甾-6(5)-烯-3β,4β,25-三醇79-1(400mg,0.96mmol)溶解于四氢呋喃(15mL)，在室温下加入三乙胺(2.9g，28.8mmol)，4-二甲氨基吡啶(117mg,0.96mmol)和乙酸酐(980mg，9.6mmol)，随后在80℃下搅拌24小时，TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，用水(50mL)淬灭，乙酸乙酯(30mL×3)萃取，有机相饱和盐水(50mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)，得到白色固体乙酸-(6R)-6-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-基酯79-2(415mg，纯度:90％，产率：71.76％)。1H 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).
第三步室温下100mL圆底烧瓶中，将乙酸-(6R)-6-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-基酯79-2(400mg,0.73mmol)溶解于丙酮(15mL)，在室温下加入N-羟基邻苯二甲酰亚(24mg，0.15mmol)，醋酸钴(13mg,0.073mmol)和叔丁基过氧化氢(265mg，2.94mmol)，随后在室温下搅拌36小时，TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，用水(50mL)淬灭，乙酸乙酯(40mL×3)萃取，有机相饱和盐水(60mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)，得到白色固体乙酸-(6R)-6-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-9a,11a-二甲基-4-氧亚基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-基酯79-3(300mg，纯度:90％，产率：65.81％)。1H 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).13C NMR(101MHz,CDCl3)δ202.20,170.06,169.47,158.31,131.14,73.61,71.51,71.12,54.68,50.80,49.91,45.95,44.37,43.06,38.53,37.93,36.42,35.80,35.69,29.31,29.27,28.52,26.20,22.21,21.15,20.99,20.78,18.83,17.91,11.92.
第四步室温下50mL圆底烧瓶中，将乙酸-(6R)-6-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基- 9a,11a-二甲基-4-氧亚基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-基酯79-3(300mg,0.54mmol)溶解于乙酸乙酯(10mL)和四氢呋喃(10mL)，在室温下加入10％Pd/C(150mg)，室温下将混合物在氢气氛围中搅拌1小时。TLC(乙酸乙酯/石油醚＝15:1)监测反应。反应结束后，用乙酸乙酯(20mL)稀释反应液，通过硅藻土过滤，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝15:1)，得白色固体乙酸-(6R)-6-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-9a,11a-二甲基-4-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-基酯79-4(200mg，纯度：90％，产率：59.60％)。1H 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,CDCl3)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.
第五步室温下50mL圆底烧瓶中，将乙酸-(6R)-6-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-9a,11a-二甲基-4-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-基酯79-4(200mg,0.36mmol)溶解于二乙胺基三氟化硫(3mL)，在40℃下搅拌6小时，TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，将反应液加入冰水(30mL)中淬灭，乙酸乙酯(20mL×3)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)，得到白色固体乙酸-(6R)-6-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6,7-二乙酰氧基-4,4-二氟-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-基酯79-5A和乙酸-(6R)-6-[(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-6,7-二乙酰氧基-4-氟-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-基酯混合物79-5B混合物(120mg，纯度:90％。以79-5A计算产率：51.55％)。
化合物79-5A:1H 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.
第六步室温下50mL圆底烧瓶中，将乙酸-(6R)-6-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-4,4-二氟-6,7-二羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-基酯79-5A和乙酸-(6R)-6-[(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-6,7-二乙酰氧基-4-氟-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-基酯混合物79-5B混合物(385mg,0.77mmol)溶解于四氢呋喃(5mL)和甲醇(5mL)，在室温下加入氢氧化锂一水合物(324mg，7.72mmol)和水(3mL)，随后在45℃下搅拌16小时，TLC(石油醚:乙酸乙酯＝2:1)监测反应，反应结束后，用水(50mL)稀释，乙酸乙酯(40mL×3)萃取，有机相饱和盐水(80mL)洗涤，无水硫 酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝2:1)，得到白色固体(1R,3aS,3bS,7S,9aR,9bS,11aR)-4,4-二氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6,7-二醇79-6A和(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-4-氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6,7-二醇79-6B的混合物(340mg，纯度:90.18％，以79-6A计算产率：86.97％)。
化合物79-6A：1H NMR(400MHz,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).
第七步室温下50mL圆底烧瓶中，将(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-4,4-二氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6,7-二醇79-6A和(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-4-氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6,7-二醇79-6B的混合物(1.41g,3.09mmol)溶解于二氯甲烷(30mL)，在室温下加入三乙胺(0.94g，9.26mmol)，4-二甲氨基吡啶(0.3g,2.47mmol)和苯甲酰氯(0.65g，4.63mmol)，随后在室温下搅拌20分钟，TLC(石油醚:乙酸乙酯＝4:1)监测反应，反应结束后，用水(100mL)淬灭，乙酸乙酯(60mL×3)萃取，有机相饱和盐水(80mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，拿到白色固体(1.5g)经手性拆分(仪器：Waters Acquity UPCC；柱：REGIS CHIRAL(S，S)-乳清O1 4.6*150mm，3.5um；流动相：A/B:CO2/MeOH(0.1％DEA)＝50/50；流速：1.5毫升/分钟；柱温度：37度)得到白色固体(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-(苄基氧基)-4,4-二氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-醇79-7A(1.04g，纯度:95％，产率：58.52％，保留时间tR＝2.597min)和(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-7-(苄基氧基)-4-氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6-醇79-7B(0.24g，纯度:95％，产率：14.02％，保留时间tR＝2.963min)。
化合物79-7A：1H NMR(400MHz,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).19F NMR(376MHz,CDCl3)δ＝-89.02(d,J＝236.3,1F),-110.79(d,J＝236.2,1F).
化合物79-7B：1H NMR(400MHz,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).19F NMR(376MHz,CDCl3)δ＝-109.82(s,1F).
第八步室温下50mL圆底烧瓶中，将(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-7-(苄基氧基)-4-氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6-醇79-7B(100mg,0.19mmol)溶解于二氯甲烷(10mL)，在室温下加入戴斯-马丁氧化剂(242mg，0.57 mmol)，在室温下搅拌2小时。TLC(乙酸乙酯/石油醚＝4：1)监测反应。反应结束后，加入饱和亚硫酸钠水溶液(5mL)和饱和碳酸氢钠水溶液(5mL)淬灭反应，二氯甲烷(10mL×3)萃取，有机相饱和盐水(10mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，得白色固体(1R,3aR,5aR,7S,9aR,9bR,11aR)-7-(苄基氧基)-4-氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6-酮79-8(81mg，纯度：80％，产率：65.05％)。1H 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＝203.65,165.72,133.24,129.90,128.38,71.12,54.36,53.16,52.49,48.80,44.37,43.60,41.83,41.17,39.43,36.44,36.19,35.61,29.41,29.37,29.22,28.49,28.34,25.12,25.03,23.37,22.28,20.84,18.97,14.97,12.61.19F NMR(376MHz,CDCl3)δ＝-110.02(s,1F)。
第九步室温下50mL圆底烧瓶中，将(1R,3aR,5aR,7S,9aR,9bR,11aR)-7-(苄基氧基)-4-氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6-酮79-8(80mg,0.15mmol)溶解于四氢呋喃(2mL)和甲醇(2mL)，在室温下加入碳酸钾(105mg，0.76mmol)和水(1mL)，随后在室温下搅拌4小时，TLC(石油醚:乙酸乙酯＝3：1)检测反应，反应结束后，用水(20mL)稀释，乙酸乙酯(10mL×3)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝3：1)，得到白色固体(1R,3aR,5aR,7S,9aR,9bR,11aR)-4-氟-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6-酮79(15.55mg，纯度：100％，产率：23.47％)。
化合物79：1H NMR(400MHz,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).13C 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
实施例85&86
化合物85或86 7β-氟-5α-胆甾-3β,25-二醇
化合物86或85 7α-氟-5α-胆甾-3β,25-二醇的制备。

第一步将(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-7(1g,2.25mmol,1eq)溶于四氢呋喃(10mL)，向溶液中加入乙酸钴(80mg,0.45mmol,0.2eq)，N-羟基临苯二甲酰亚胺(0.15g,0.90mmol,0.4eq)，过氧化叔丁醇(1.01g,11.25mmol,5eq)，室温搅拌20小时。反应完成后，加入30mL水，用乙酸乙酯(30mL*3)萃取，将得到的有机相用无水硫酸钠干燥后旋干得到粗品，粗品经硅胶过柱(石油醚/乙酸乙酯＝10/1)纯化得到(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-4-氧亚基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯72-1(0.6g,1.31mmol,收率：58％)。1H NMR(400MHz,CDCl3)δ5.70(d,J＝1.5Hz,1H),4.80–4.64(m,1H),3.67(s,3H),2.48(dddd,J＝15.6,12.1,9.1,2.4Hz,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).
第二步将化合物(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-4-氧亚基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯72-1(0.6g,1.31mmol,1eq)溶于乙酸乙酯(15mL)中,然后向其中加入钯碳(10％)(120mg),室温搅拌12小时后。过滤,旋干有机溶剂得到产物(5R)-5-[(1R,3aS,3bR,7S,9aS,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-4-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯72-2(450mg,0.977mmol,收率：75％)为油状化合物。1H NMR(400MHz,cdcl3)δ5.36–5.24(m,1H),4.30(s,3H),3.00–2.81(m,5H),2.70–2.58(m,5H),2.55–2.47(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).
第三步将(5R)-5-[(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-4-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯72-2(300mg,0.65mmol,1eq)溶于甲醇(10mL)，向溶液中加入硼氢化钠(30mg,0.78mmol,1.2eq)，室温搅拌1小时。反应完成后，加入30mL水，用乙酸乙酯(30mL*3)萃取，将得到的有机相用无水硫酸钠干燥后旋干得到粗品为(5R)-5-[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-7-乙酰氧基-4-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯85-1(240mg,0.52mmol,收率：80％)，直接投下一步。
第四步将化合物(5R)-5-[(1R,3aS,3bR,5aS,7S,9aS,9bS,11aR)-7-乙酰氧基-4-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯85-1(240mg,0.52mmol,1eq)溶于超干吡啶(5mL)中,然后向其中加入4-甲基苯磺酰氯(824mg,4.32mmol,10eq)，室温搅拌5天,反应完成后，加入30ml 3M盐酸水溶液，用乙酸乙酯(30mL*3)萃取，然后旋干有机溶剂得到粗品，粗品经过硅胶分离纯化(石油醚:乙酸乙酯＝10:1)得到产物经过SFC拆分(仪器：Waters Acquity UPCC；色谱柱：Daicel CHIRALPAK IB 4.6*250mm，5um；流动相：A/B:CO2/MeOH(0.1％DEA)＝60/40；流速：1.5毫升/分钟；柱温度：37度)得到85-2(60mg,0.1mmol,收率：22.5％，保留时间tR＝1.769min)和86-1(70mg,0.13mmol,收率：30％，保留时间tR＝2.792min)为油状化合物。
化合物85-2：1H NMR(400MHz,CDCl3)δ7.76(d,J＝8.1Hz,1H),7.31(d,J＝8.1Hz,1H),4.67–4.57(m,0H),4.46(td,J＝10.4,5.1Hz,0H),3.67(s,3H),2.44(s,3H),2.26(dd,J＝15.3,8.2Hz,2H),2.00(s,3H),1.96(d,J＝13.0Hz,1H),1.81(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).
化合物86-1：1H NMR(400MHz,CDCl3)δ7.78(d,J＝8.1Hz,2H),7.35(d,J＝8.0Hz,2H),4.76(s,1H),4.62–4.46(m,1H),3.68(s,3H),2.45(s,3H),2.26(dt,J＝15.7,7.7Hz,2H),2.00(s,3H),1.90(d,J＝13.0Hz,1H),1.79(d,J＝13.2Hz,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).
第五步将85-2(60mg,0.1mmol,1.0eq),TBAF(38mg,0.146mmol,1.5eq)溶于四氢呋喃(2mL)中，65℃搅拌12小时,TLC检测,反应完成后向其中加入水摧灭,然后用乙酸乙酯(50mL)萃取,有机相用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,过滤,旋干得到粗品,然后经过硅胶分离纯化(石油醚:乙酸乙酯＝1:1)得到85-3(20mg,0.145mmol,收率：44％)，直接往后投下一步
第六步将85-3(20mg,0.043mmol,1eq)溶于四氢呋喃(2mL)中，置换氮气后缓慢滴加甲基溴化镁(2.4M)(0.6mL，10eq)，室温搅拌3小时,TLC检测,反应完成后向其中加入水摧灭,然后用乙酸乙酯(50mL)萃取,有机相用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,过滤,旋干得到粗品,然后经过硅胶分离纯化(石油醚:乙酸乙酯＝4:1)得到85(6mg,0.014mmol,收率：33％)
化合物85：1H NMR(400MHz,CDCl3)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,6H),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).19F NMR(376MHz,CDCl3)δ-196.28.
类似的操作将原料85-2换成86-1得到86。
化合物86：1H NMR(400MHz,CDCl3)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,CDCl3)δ-167.50.
实施例94
化合物94(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4,4-二氟-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6,7-二醇的制备

第一步室温下，将(R)-5-((3S，4R，8S，9S，10R，13R，14S，17R)-3-乙酰氧基-4-羟基-10,13-二甲基-2,3,4,7,8,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊烷[a]菲17-基)-2,2-二氟己酸乙酯34-1(450mg,0.88mmol)溶解于无水四氢呋喃(12mL)，氮气保护下，降温到0℃，滴加甲基溴化镁(3M in 2-Me-THF，2.9mL，8.8mmol)，随后转至室温搅拌2小时，TLC(石油醚:乙酸乙酯＝2：1)监测反应，反应结束后，体系降温至0℃，用饱和氯化铵水溶液(30mL)淬灭，乙酸乙酯(30mL×2)萃取，有机相饱和盐水(50mL)洗涤，无水硫酸钠干燥，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，得到白色固体(3S，4R，8S，9S，10R，13R，14S，17R)-17-((R)-5,5-二氟-6-羟基-6-甲基庚烷-2-基)-10,13-二甲基-2,3,4,7,8,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊二烯[a]菲-3,4-二醇94-1(195mg，纯度:95％，产率：49％)。1H NMR(400MHz,CDCl3)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),1.68–1.53(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).
第二步室温下将3S，4R，8S，9S，10R，13R，14S，17R)-17-((R)-5,5-二氟-6-羟基-6-甲基庚烷-2-基)-10,13-二甲基-2,3,4,7,8,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊二烯[a]菲-3,4-二醇94-1(193mg,0.42mmol，1.0eq)溶解于四氢呋喃(10mL)，在室温下加入三乙胺(6.37g，63.0mmol,150eq)，4-二甲氨基吡啶(154mg,1.26mmol,3.0eq)和乙酸酐(2.14g，21.0mmol,50eq)，随后在80℃下搅拌24小时，TLC(石油醚:乙酸乙酯＝1:1)监测反应。反应结束后，用冰水(50mL)淬灭，乙酸乙酯(30mL×2) 萃取，有机相饱和盐水(50mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝20：1)，得到白色固体(3S，4R，8S，9S，10R，13R，14S，17R)-17-((R)-6-乙酰氧基-5,5-二氟-6-甲基庚烷-2-基)-10,13-二甲基-2,3,4,7,8,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊二烯[a]菲-3,4-二基二乙酸酯94-2(182mg，纯度:90％，产率：74.59％)。
1H NMR(400MHz,CDCl3)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).
第三步室温下，将(3S，4R，8S，9S，10R，13R，14S，17R)-17-((R)-6-乙酰氧基-5,5-二氟-6-甲基庚烷-2-基)-10,13-二甲基-2,3,4,7,8,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊二烯[a]菲-3,4-二基二乙酸酯94-2(182mg,0.31mmol，1.0eq)溶解于丙酮(8mL)，在室温下加入N-羟基邻苯二甲酰亚(10mg，0.062mmol，0.2eq)，醋酸钴(5.5mg,0.031mmol，0.1eq)和叔丁基过氧化氢(5M in hexane，0.25mL，1.24mmol，4.0eq)，随后在室温下搅拌36小时，TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，用水(30mL)淬灭，乙酸乙酯(20mL×2)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝15：1)，得到白色固体(3S，4R，8S，9S，10R，13R，14S，17R)-17-((R)-6-乙酰氧基-5,5-二氟-6-甲基庚烷-2-基)-10,13-二甲基-7-氧基-2,3,4,7,8,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊[a]菲-3,4-二基二乙酸酯94-3(120mg，纯度:90％，产率：65.22％)。1H NMR(400MHz,CDCl3)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).19F NMR(376MHz,CDCl3)δ-113.92–-117.67(m,2F).
第四步室温下，将(3S，4R，8S，9S，10R，13R，14S，17R)-17-((R)-6-乙酰氧基-5,5-二氟-6-甲基庚烷-2-基)-10,13-二甲基-7-氧基-2,3,4,7,8,9,10,11,12,13,14,15,16,17-十四氢-1H-环戊[a]菲-3,4-二基二乙酸酯94-3(120mg,0.20mmol)溶解于乙酸乙酯(3mL)和四氢呋喃(3mL)，在室温下加入10％Pd/C(60mg)，室温下将混合物在氢气氛围中搅拌6小时。TLC(乙酸乙酯/石油醚＝1:4)检测反应。反应结束后，用乙酸乙酯(20mL)稀释反应液，通过硅藻土过滤，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝15:1)，得到白色固体(3S，4R，5R，8S，9S，10R，13R，14S，17R)-17-((R)-6-乙酰氧基-5,5-二氟-6-甲基庚烷-2-基)-10,13-二甲基-7-氧代十六氢-1H-环戊二烯[a]菲-3,4-二基二乙酸酯94-4(53mg，纯度：90％，产率：44.54％)。1H NMR(400MHz,CDCl3)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,3H),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).
第五步室温下，将(3S，4R，5R，8S，9S，10R，13R，14S，17R)-17-((R)-6-乙酰氧基-5,5-二氟-6-甲基庚烷-2-基)-10,13-二甲基-7-氧代十六氢-1H-环戊二烯[a]菲-3,4-二基二乙酸酯94-4(53mg,0.089mmol)溶解于二乙胺基三氟化硫(2mL)，在70℃下搅拌2小时，TLC(石油醚:乙酸乙酯＝4:1)监测反应，反应结束后，将反应液乙酸乙酯稀释，并慢慢滴加到冰水(20mL)中淬灭，乙酸乙酯(15mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)，得到白色固体(3S，4R，5R，8S，9S，10R，13R，14S，17R)-17-((R)-6-乙酰氧基-5,5-二氟-6-甲基庚烷-2-基)-7,7-二氟-10,13-二甲基十六氢-1H-环戊二烯[a]菲-3,4-二基二乙酸酯94-5(45mg，纯度:90％，产率：81.82％)。1H NMR(400MHz,CDCl3)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,CDCl3)δ-89.25(d,J＝238.2Hz,1F),-110.61(d,J＝237.8Hz,1F),-114.63–-116.99(m,2F).
第六步室温下，将(3S，4R，5R，8S，9S，10R，13R，14S，17R)-17-((R)-6-乙酰氧基-5,5-二氟-6-甲基庚烷-2-基)-7,7-二氟-10,13-二甲基十六氢-1H-环戊二烯[a]菲-3,4-二基二乙酸酯94-5(45mg,0.073mmol，1.0eq)溶解于四氢呋喃(1mL)和甲醇(1mL)，在室温下加入氢氧化锂一水合物(15mg，0.365mmol，5.0eq)和水(0.5mL)，随后在室温下搅拌30min，TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，用水(10mL)稀释，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝2:1)，得到白色固体(3S，4R，5R，8S，9S，10R，13R，14S，17R)-17-((R)-5,5-二氟-6-羟基-6-甲基庚烷-2-基)-7,7-二氟-10,13-二甲基十六氢-1H-环戊二烯[a]菲-3,4-二醇94-6(28mg，纯度:90％，产率：77.78％)。1H NMR(400MHz,CDCl3)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).
第七步室温下，将(3S，4R，5R，8S，9S，10R，13R，14S，17R)-17-((R)-5,5-二氟-6-羟基-6-甲基庚烷-2-基)-7,7-二氟-10,13-二甲基十六氢-1H-环戊二烯[a]菲-3,4-二醇94-6(28mg,0.057mmol，1.0eq)溶解于二氯甲烷(2mL)和四氢呋喃(1mL)，在室温下加入三乙胺(35mg，0.34mmol，6.0eq)，4-二甲氨基吡啶(7mg,0.057mmol，1.0eq)和苯甲酰氯(24mg，0.17mmol，3.0eq)，随后在室温下搅拌8小时，TLC(石油醚:乙酸乙酯＝1:1)监测反应，反应结束后，用水(10mL)淬灭，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)和SFC拆分(仪器：Waters Acquity UPCC；色谱柱：Daicel CHIRALPAK IB 4.6*250mm，5um；流动相：A/B:CO2/MeOH(0.1％DEA)＝60/40；流速：1.5毫升/分钟；柱温度：37 度)得到白色固体(3S，4R，5R，8S，9S，10R，13R，14S，17R)-17-((R)-5,5-二氟-6-羟基-6-甲基庚烷-2-基)-7,7-二氟-4-羟基-10,13-二甲基十六氢-1H-环戊基[a]菲-3-基苯甲酸酯94-7(15mg，纯度:95％，产率：44.12％,保留时间tR＝1.225min)。1H NMR(400MHz,CDCl3)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).
第八步室温下，将(3S，4R，5R，8S，9S，10R，13R，14S，17R)-17-((R)-5,5-二氟-6-羟基-6-甲基庚烷-2-基)-7,7-二氟-4-羟基-10,13-二甲基十六氢-1H-环戊基[a]菲-3-基苯甲酸酯94-7(15mg,0.025mmol，1.0eq)溶解于四氢呋喃(0.5mL)和甲醇(0.5mL)，在室温下加入氢氧化锂一水合物(11mg，0.25mmol，10.0eq)和水(0.2mL)，随后在室温下搅拌30min，TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，用水(10mL)稀释，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝4:1)，得到白色固体(3S，4R，5R，8S，9S，10R，13R，14S，17R)-17-((R)-5,5-二氟-6-羟基-6-甲基庚烷-2-基)-7,7-二氟-10,13-二甲基十六氢-1H-环戊二烯[a]菲-3,4-二醇94(6.23mg，纯度:100％，产率：50.89％)。
化合物94：1H NMR(400MHz,CDCl3)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)。19F NMR(377MHz,CDCl3)δ-88.97(d,J＝235.8Hz,1F),-110.95(d,J＝235.7Hz,1F),-114.05–-117.90(m,2F).
实施例103
化合物103 3β,25-二羟基-5α-胆甾-7-甲腈的制备

第一步将乙酸-(1R,3aS,3bR,7S,9aS,9bS,11aR)-4-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯(200mg,0.432mmol,1eq)溶于超干吡啶(5mL)中,然后向其中加入4-甲基苯磺酰氯(824mg,4.32mmol,10eq)，室温搅拌5天,反应完成后，加入30ml 3M盐酸水溶液，用乙酸乙酯(30mL*3)萃取，然后旋干有机溶剂得到粗品，粗品经过硅胶分离纯化(石油醚:乙酸乙酯＝10:1)得到产物4-甲基苯磺酸-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-7-乙酰氧基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-4-基酯(30mg,0.049mmol,收率：11％)为白色化合物。
1H NMR(400MHz,CDCl3)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,5H),0.90(d,J＝6.4Hz,3H),0.79(d,J＝15.5Hz,3H),0.62(d,J＝7.6Hz,3H).
第二步将化合物4-甲基苯磺酸-(1R,3aS,3bR,5aR,7S,9aS,9bS,11aR)-7-乙酰氧基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-4-基酯103-1(60mg,0.1mmol,1.0eq),氰化钠(20mg,0.4mmol,4eq)溶于DMF(2mL)中，65℃搅拌12小时,TLC(石油醚:乙酸乙酯＝1:1)监测,反应完成后向其中加入水淬灭,然后用乙酸乙酯(50mL)萃取,有机相用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,过滤,旋干得到粗品,然后经过硅胶分离纯化(石油醚:乙酸乙酯＝1:1)得到乙酸-(1R,3aS,3bR,7S,9aS,9bS,11aR)-4-氰基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯103-2(25mg,0.05mmol,收率：55％)，粗品直接投入下一步。
第三步将化合物乙酸-(1R,3aS,3bR,7S,9aS,9bS,11aR)-4-氰基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯103-2(25mg,0.05mmol,1eq)溶于甲醇(2mL)中，加入碳酸钾(29mg,0.212mmol,4eq)，室温搅拌3小时,TLC(石油醚:乙酸乙酯＝4:1)监测,反应完成后向其中加入水淬灭,然后用乙酸乙酯(50mL)萃取,有机相用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,过滤,旋干得到粗品,然后经过硅胶分离纯化(石油醚:乙酸乙酯＝4:1)得到3β,25-二羟基胆甾-7-甲腈103(16mg,0.037mmol,收率：70％)。
化合物103：1H NMR(400MHz,CDCl3)δ3.67(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＝6.5Hz,3H),0.81(d,J＝4.2Hz,3H),0.66(s,3H).13C NMR(101MHz,CDCl3)δ121.18,71.10,70.63,55.76,52.83,49.11,44.38,42.74,40.40,39.01,37.16,36.36,36.28,35.61,35.49,32.36,31.14,30.80,29.29,29.27,28.04,23.60,20.89,20.66,18.57,12.03,11.92.
实施例106
化合物106 24-[羟基(2-甲氧基苯基)甲基]-7-甲亚基-5α-胆烷-3β,4β-二醇的制备
第一步：将原料1-溴-2-甲氧基苯(7.64g,40.82mmol,3.5eq)溶解在无水四氢呋喃(50mL)中，体系冷却至-78℃，加入正丁基锂(14.0mL,34.99mmol,3.0eq)，并保持温度搅拌30min。将化合物(R)-5-(((3aS，5aR，5bS，7aR，8R，10aS，10bS，12bR)-2,2,5a，7a-四甲基-3a，5,5a，5b，6,7,7a，8,9,10,10a，10b，11,12b-四氢-4H-环戊基[7,8]菲[1,2-d][1,3]二氧基-8-基)己醛III(5g,11.66mmol，1.0eq)溶解在四氢呋喃(5mL)中加入到上述反应液中，反应体系在-78℃搅拌30min。TLC(石油醚:乙酸乙酯＝5:1)监测反应进度，原料消耗完毕。将10mL水加入到反应体系中，使用乙酸乙酯(100mL×3)萃取，合并有机相用饱和氯化钠(100mL)洗涤，合并有机相干燥，浓缩得到粗品。粗品通过硅胶柱层析(石油醚:乙酸乙酯＝90：10)得到白色固体(5R)-5-[(3aS，5aR，5bS，7aR，8R，10aS，10bS，12bR) -2，2，5a，7a-四甲基-4，5，5a，5b，6，7，7a，8，9，10，10a，10b，11，12b-四氢-3aH-环戊二烯[1’，2’：1,2]菲[7，8-d][1,3]二氧杂环-8-基]-1-(2-甲氧基苯基)己-1-醇106-1(4.9g,纯度90％，收率70.43％)。1H NMR(400MHz,CDCl3)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).
第二步：室温下在200mL的圆底烧瓶中，将(5R)-5-[(3aS，5aR，5bS，7aR，8R，10aS，10bS，12bR)-2，2，5a，7a-四甲基-4，5，5a，5b，6，7，7a，8，9，10，10a，10b，11，12b-四氢-3aH-环戊二烯[1’，2’：1,2]菲[7，8-d][1,3]二氧杂环-8-基]-1-(2-甲氧基苯基)己-1-醇106-1(4.9g,9.13mmol)溶解于二氯甲烷(70mL)，在室温下加入叔丁基二甲基氯硅烷(4.13g，27.39mmol)，4-二甲氨基吡啶(450mg,3.65mmol)和咪唑(2.49g，36.51mmol)，随后在室温下搅拌2小时。TLC(石油醚:乙酸乙酯＝5:1)监测反应，反应结束后，用水(100mL)稀释，乙酸乙酯(100mL×3)萃取，有机相饱和盐水(50mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)，得到白色固体(9R)-9-[(3aS，5aR，5bS，7aR，8R，10aS，10bS，12bR)-2，2，5a，7a-四甲基-4，5，5a，5b，6，7，7a，8，9，10，10a，10b，11，12b-四氢-3aH-环戊二烯[1’，2’：1,2]菲[7，8-d][1,3]二氧杂环-8-基]-5-(2-甲氧基苯基)-2，3，3-四甲基-4-氧-3-硅癸烷106-2(4.9g，纯度:90％，产率：74％)。1H 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,3H),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).
第三步：室温下250mL圆底烧瓶中，将(9R)-9-[(3aS，5aR，5bS，7aR，8R，10aS，10bS，12bR)-2，2，5a，7a-四甲基-4，5，5a，5b，6，7，7a，8，9，10，10a，10b，11，12b-四氢-3aH-环戊二烯[1’，2’：1,2]菲[7，8-d][1,3]二氧杂环-8-基]-5-(2-甲氧基苯基)-2，3，3-四甲基-4-氧-3-硅癸烷106-2(4.9g,7.53mmol)溶解于丙酮(100mL)，在室温下加入N-羟基邻苯二甲酰亚(370mg，2.26mmol)，醋酸钴(200mg,1.13mmol)和叔丁基过氧化氢(3.39g，37.63mmol)，随后在室温下搅拌36小时.TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，用水(50mL)淬灭，乙酸乙酯(100mL×3)萃取，有机相饱和盐水(60mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)，得到白色固体(3aS，5aR，5bS，7aR，8R，10aS，10bS，12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧-3-硅烷基]-2,2,5a，7a，7a-四甲基-4,5,5a，5b，6,7,8,9,10,10a，10b，11,12b-四氢-3aH-环戊烯[1':1,2]菲[7,8-d][1,3]二氧基-11,5-酮106-3(2.6g，纯度:90％，产率：46.75％)。1H NMR(400MHz, 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).
第四步：将(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12b-十四氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-11-酮106-3(2.1g,3.158mmol)溶于甲醇(30mL)，加入Pd/C(1.08g,5.052mmol)，将体系置换成氢气，反应液在室温下搅拌2小时。反应液经硅藻土过滤，滤液旋干经硅胶柱层析纯化，得到(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-酮106-4(1.1g,1.402mmol,44.39％)为白色固体。1H 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＝11.1,8.1Hz,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).
第五步：将甲基三苯基溴化磷(696.19mg,1.949mmol)置于两颈烧瓶(50mL)，置换成氮气后加入超干四氢呋喃(15mL)，在0℃下搅拌并逐滴加入在四氢呋喃中的叔丁醇钾(1.949mL,1.949mmol)0℃下继续搅拌10分钟。将(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-酮106-4(260mg,0.390mmol)溶在超干四氢呋喃(5mL)中，逐滴加入到反应瓶中，反应混合液在室温下搅拌1小时。用5％的磷酸二氢钠溶液猝灭反应，加乙酸乙酯萃取，有机相经硅胶柱层析(石油醚/二氯甲烷＝2/1)纯化，得到(9R)-9-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-11-甲亚基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸烷106-5(210mg,0.253mmol,64.81％)为无色透明油。1H 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,4H),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).
第六步：将(9R)-9-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-11-甲亚基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸烷106-5(18mg,0.027mmol)溶于四氢呋喃(1mL)，加入盐酸(2.520mL,7.560mmol)，40℃下搅拌3小时，TLC(石油醚:乙酸乙酯＝1:1)监测反应，原料反应完。向反应液加入水，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，旋干。粗品经硅胶柱层析(PE/EA＝1/1)纯化，得到24-[羟基(2-甲氧基苯基)甲基]-7-甲亚基-5α-胆烷-3β,4β-二醇106(6mg,0.011mmol,41.24％)为白色固体。
化合物106：1H NMR(400MHz,CDCl3)δ7.30(d,J＝5.8Hz,1H),7.24(d,J＝7.8Hz,1H),6.97(dd,J＝15.5,8.1Hz,1H),6.88(d,J＝8.3Hz,1H),4.91–4.80(m,1H),4.65(d,J＝42.3Hz,2H),3.86(s,3H),3.79(s,1H),3.61–3.49(m,1H),2.49(t,J＝12.8Hz,1H),2.09–1.92(m,5H),1.84(dd,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
实施例109
化合物109 3β,25-二羟基-5α-胆甾-4-甲腈的制备

第一步：室温下50mL圆底烧瓶中，将3β-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-25-羟基胆甾-6(5)-烯-4-甲醛23-3(90mg,0.17mmol)溶解于乙腈(6mL)，在室温下加入乙酸钠(230mg，2.8mmol)，盐酸羟胺(115mg，1.7mmol)和水(2ml)，随后在室温下搅拌30小时，TLC(石油醚:乙酸乙酯＝4:1)监测反应，反应结束后，用水(20mL)淬灭，乙酸乙酯(20mL×3)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，得到白色固体[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6-基]甲烷醛肟109-1(53mg，纯度:90％，产率：51.58％)。1H NMR(400MHz,CDCl3)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).
第二步：室温下50mL圆底烧瓶中，将[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6-基]甲烷醛肟109-1(43mg,0.077mmol)溶解于四氢呋喃(2.5mL)，在室温下加入二苯二硫醚(67mg，0.31mmol)，三丁基膦(62mg，0.31mmol)，随后在室温下搅拌6小时，TLC(石油醚:乙酸乙酯＝8:1)监测反应，反应结束后，用水(10mL)淬灭，乙酸乙酯(10mL×3)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝8：1)，得到白色固体3β-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-25-羟基胆甾-4-甲腈109-2(31mg，纯度:90％，产率：67.04％)。1H NMR(400MHz,CDCl3)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).13C NMR(101MHz,CDCl3)δ142.57,119.68,71.12,70.74,56.45,56.12,55.12,45.76,44.41,42.55,40.84,39.80,36.60,36.41,36.14,35.71,35.19,31.90,31.52,29.71,29.40,29.20,29.07,28.22,27.08,25.71,24.17,20.72,18.64,18.02,13.09,12.05.
第三步：室温下50mL圆底烧瓶中，将3β-{[二甲基(2-甲基丙-2-基)甲硅基]氧基}-25-羟基胆甾-4-甲腈109-2(31mg,0.057mmol)溶解于一摩尔每升的四丁基氟化铵四氢呋喃溶液(2mL)，在室温下搅拌2小时，TLC(石油醚:乙酸乙酯＝4:1)监测反应，反应结束后，用水(10mL)淬灭，乙酸乙酯(10mL×3)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，得到白色固体3β,25-二羟基-5α-胆甾-4-甲腈109(18.02mg，纯度:100％，产率：73.59％)化合物109：1H NMR(400MHz,CDCl3)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).13C NMR(101MHz,CDCl3)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+H-H2O]+＝412.5。
实施例114
化合物114 24-[羟基(2-甲氧基苯基)甲基]-7-(羟基甲基)-5α-胆烷-3β,4β-二醇的制备
第一步：将(9R)-9-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-11-甲亚基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸烷106-5(180mg,0.271mmol)溶于超干四氢呋喃(5mL)，置换成氮气，在冰水浴下加入硼烷二甲基硫醚(0.406mL,0.812mmol)。混合溶液在室温下搅拌2小时。随后向反应液中滴加氢氧化钠水溶液(0.162mL,1.624mmol)和过氧化氢(184.09mg,1.624mmol)。混合液在室温下搅拌过夜。反应液用亚硫酸氢钠水溶液猝灭，加乙酸乙酯萃取，有机相用无水硫酸钠干燥，旋干。粗品经硅胶柱层析(石油醚/乙酸乙酯＝3/1)纯化，得到 [(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-11-基]甲醇114-1(90mg,0.112mmol,41.38％)为白色固体。1H 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,2H),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).
第二步：将[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-11-基]甲醇114-1(20mg,0.029mmol)溶于四氢呋喃(1mL)中，加入盐酸(2mL,6.000mmol)，混合液在室温下搅拌2小时。向反应液加入水，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，旋干。粗品经硅胶柱层析(石油醚:乙酸乙酯＝1:4)纯化，得到24-[羟基(2-甲氧基苯基)甲基]-7-(羟基甲基)-5α-胆烷-3β,4β-二醇白色固体114(4mg,0.007mmol,23.26％)
化合物114：1H NMR(400MHz,CDCl3)δ7.27(s,1H),7.25(s,1H),6.96(s,1H),6.89(d,J＝8.2Hz,1H),4.83(d,J＝7.0Hz,1H),3.86(s,3H),3.77(d,J＝6.9Hz,1H),3.73(s,1H),3.67(t,J＝10.6Hz,1H),3.58(d,J＝10.9Hz,1H),1.88(d,J＝12.5Hz,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,3H),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-2H2O]+＝493.45。
实施例118
化合物118 4-(羟基甲基)-5α-胆甾-3β,25-二醇的制备

第一步：将化合物(5R)-5-[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-8(150mg,0.326mmol,1.0eq)溶解在乙酸乙酯(5mL)中,加入二氧化铂(36.97mg,0.163mmol,0.5eq)，所得混合液在反应液在H2，25℃中搅拌3h。TLC(石油醚:乙酸乙酯＝3:1)监测反应进度，原料消耗完毕。过滤，滤液浓缩得到粗品，粗品硅胶柱层析纯化(石油醚：乙酸乙酯：80：20)，得到白色固体(5R)-5-[(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯IV-1(100mg,0.195mmol,59.74％)。1H 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).
第二步：在50mL圆底烧瓶中，将(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-6-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯IV-1(200mg,0.43mmol)溶解于二氯甲烷(10mL)，在室温下加入N-甲基吗啉氧化物(76mg，0.65mmol),四丙基高钌酸铵(30mg,0.086mmol)和四A分子筛(190mg,0.43mmol),将混合物在室温下搅拌24小时。TLC(石油醚/乙酸乙酯＝10/1)监测反应。反应结束后，向反应液加入水(50mL)中，用乙酸乙酯(30mL×3)萃取，用饱和食盐水(50mL)洗涤萃取的有机溶液，用无水硫酸钠干燥，过滤，浓缩后得粗产品。粗产品经硅胶柱层析纯化分离(洗脱剂：石油醚/乙酸乙酯＝10/1)，得到白色固体(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-6-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯118-1(170mg，纯度：90％，产率：76.83％)。1H NMR(400MHz,CDCl3)δ5.16(dd,J＝12.0,7.5Hz,1H),3.67(s,3H),2.31–2.18(m,4H),2.15(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＝205.71,174.32,170.18,76.20,57.39,56.06,55.90,54.24,51.44,42.67,42.55,39.80,35.93,35.48,35.36,34.91,34.50,30.23,28.50,28.12,24.10,21.70,21.52,20.75,20.17,18.54,13.69,12.03.
第三步：在50mL圆底烧瓶中，室温下将甲基三苯基溴化膦(388mg，1.09mmol)悬浮于四氢呋喃(4mL)，在0℃下逐滴加入1M/L的叔丁醇钾四氢呋喃溶液(1.09mL，1.09mmol),将混合物在室温下搅拌2小时后,在0℃下向反应液中逐滴加入(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-6-氧亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯118-1(100mg,0.22mmol)的四氢呋喃(1mL)溶液,将混合物在室温下搅拌5小时。TLC(石油醚/乙酸乙酯＝10/1)监测反应。反应结束后，向反应液加入水(30mL)中，用乙酸乙酯(20mL×3)萃取，用饱和食盐水(40mL)洗涤萃取的有机溶液，用无水硫酸钠干燥，过滤，浓缩后得粗产品。粗产品经硅胶柱层析纯化分离(洗脱剂：石油醚/乙酸乙酯＝10/1)，得到白色固体(5R)-5-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-6-甲亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯118-2(64mg，纯度：90％，产率：57.85％)。1H 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.
第四步：室温下在50mL的圆底烧瓶中，将(5R)-5-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-6-甲亚基十六氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯118-2(68mg,0.15mmol)溶解于无水四氢呋喃(4mL)，反应氮气保护下，降温到0℃，滴加三摩尔每升的甲基溴化镁四氢呋喃溶液(0.50mL，1.50mmol)，随后转至室温搅拌2小时，TLC(石油醚:乙酸乙酯＝4：1)监测反应，反应结束后，体系降温至0℃，用饱和氯化铵水溶液(15mL)淬灭，乙酸乙酯(10mL×3)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，得到白色固体4-甲亚基胆甾-3β,25-二醇118-3(34mg，纯度:90％，产率：46.46％)。1H 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)δ152.30,151.31,102.19,101.34,72.35,71.24,70.11,55.38,55.24,55.20,55.13,54.36,53.48,53.45,48.86,43.40,41.58,41.50,39.04,38.94,38.63,37.58,36.75,36.23,35.39,34.72,34.22,33.99,32.02,30.56,29.46,28.68,28.34,28.32,28.19,27.25,23.23,23.18,23.13,22.44,21.67,20.69,19.98,19.75,17.61,13.11,12.30,11.91,11.05,11.04.
第五步：在50mL圆底烧瓶中，室温下将4-甲亚基胆甾-3β,25-二醇118-3(34mg,0.082mmol)溶于四氢呋喃(4mL)，在0℃下逐滴加入二摩尔每升硼烷二甲硫醚络合物的四氢呋喃溶液(0.12mL，0.24mmol),将混合物在室温下搅拌3小时后,在0℃下向反应液中逐滴加入十摩尔每升氢氧化钠水溶液(0.049mL,0.49mmol)和百分之三十的过氧化氢水溶液(56mg,0.49mmol),将混合物在室温下搅拌16小时.TLC(石油醚/乙酸乙酯＝2/1)监测反应。反应结束后，向反应液加入水(10mL)中，用乙酸乙酯(10mL×3)萃取，用饱和食盐水(20mL)洗涤萃取的有机溶液，用无水硫酸钠干燥，过滤，浓缩后得粗产品。 粗产品经硅胶柱层析纯化分离(洗脱剂：石油醚/乙酸乙酯＝2/1)，得到白色固体4-(羟基甲基)-5α-胆甾-3β,25-二醇118(22.27mg，纯度：73.65％，产率：46.05％)。1H NMR(400MHz,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).13C 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-2H2O]+＝399
实施例13
化合物13乙酸-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]甲基酯和乙酸-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-乙酰氧基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]甲基酯的制备
第一步室温下50mL圆底烧瓶中，将4-(羟基甲基)-5α-胆甾-3β,25-二醇118(180mg,0.41mmol)溶解于二氯甲烷(2mL)和四氢呋喃(2mL)，在室温下加入三乙胺(126mg，1.24mmol)，4-二甲氨基吡啶(5mg,0.041mmol)和乙酸酐(42mg，0.41mmol)，随后在室温下搅拌1小时，TLC(石油醚:乙酸乙酯＝3:1)监测反应。反应结束后，用水(30mL)淬灭，乙酸乙酯(20mL×3)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝3：1)，得到白色固体乙酸-[(1R,3aS,3bS,5aS,7S,9aR,9bS,11aR)-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]甲基酯13(180mg，纯度:90％，产率：82.06％)。
化合物13：1H NMR(400MHz,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.
实施例119
化合物119 4β-羟基-3β-羟基-24-[羟基(2-甲氧基苯基)甲基]-5α-胆烷-7-甲酸的制备
第一步：将[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-11-基]甲醇114-1(70mg,0.102mmol)溶于二氯甲烷99.9％(3mL)，加入N-甲基吗啉氧化物(21mg,0.179mmol)、四丙基高钌酸铵(14mg,0.040mmol)和分子筛。混合物在室温下搅拌30分钟。加入水，用乙酸乙酯萃取，有机相用饱和食盐水洗涤并用无水硫酸钠干燥，旋干。粗品经硅胶柱层析(石油醚/乙酸乙酯＝4/1)纯化，得到(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-甲醛119-1(60mg,0.075mmol,73.08％)为白色固体。1H NMR(400MHz,CDCl3)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),2.59(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).
第二步：将(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-甲醛119-1(55mg,0.081mmol)溶于叔丁醇(1mL)中，在冰水浴下加入2-甲基-2-丁烯(33.98mg,0.485mmol)，搅拌1分钟。随后加入亚氯酸钠(25.56mg,0.283mmol)水溶液、磷酸二氢钠(2.62mg,0.022mmol)水溶液。混合液移至室温搅拌1小时。向反应液中加入水，用乙酸乙酯萃取，有机相用饱和氯化钠溶液洗涤并用无水硫酸钠干燥，旋干。粗品经硅胶柱层析(石油醚/乙酸乙酯＝3/1)纯化得到(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-甲酸119-2(50mg,0.062mmol,76.39％)为白色固体。1H NMR(400MHz,CDCl3)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),3.99(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＝13.9,11.3Hz,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).
第三步：将(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-甲酸119-2(50mg,0.072mmol)溶于四氢呋喃(2mL)，加入盐酸(1mL,3.000mmol)。混合液在40℃下搅拌2.5小时。向反应液中加入水，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，旋干。粗品经快速色谱仪(石油醚/乙酸乙酯＝90％-100％)纯化得到4β-羟基-3β-羟基-24-[羟基(2-甲氧基苯基)甲基]-5α-胆烷-7-甲酸119(11.5mg,0.015mmol,20.68％)为白色固体。
化合物119：1H 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＝11.1Hz,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+H-H2O]+＝525.40.
实施例121&122&125
化合物121：[(1R,3aS,3bS,5aS,7S,9aS,9bS,11aR)-4,4-二氟-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]- 9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]乙腈，
化合物122：(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-4,4-二氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-6-甲亚基十六氢-1H-环戊并[1,2-a]菲-7-醇和化合物125：(1R,3aS,3bS,5aS,7S,9aS,9bS,11aR)-4,4-二氟-6-(羟基甲基)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-醇的制备
第一步：室温下50mL圆底烧瓶中，将(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-7-(苄基氧基)-4,4-二氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-醇79-7A(790mg,1.445mmol)溶解于1,2-二氯乙烷(60mL)，在室温下加入戴斯-马丁氧化剂(1838.04mg，4.335mmol)，在室温下搅拌2小时。TLC(乙酸乙酯/石油醚＝4：1)监测反应。反应结束后，加入饱和亚硫酸钠水溶液(30mL)和饱和碳酸氢钠水溶液(30mL)淬灭反应，二氯甲烷(50mL×3)萃取，有机相饱和盐水(80mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，得白色固体 (1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-(苄基氧基)-4,4-二氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-酮121-1(750mg，纯度：90％，产率：85.76％)。1H 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)。19F NMR(376MHz,CDCl3)δ＝-90.57(d,J＝240.1,1F),-111.72(d,J＝240.0,1F).
第二步：在50mL圆底烧瓶中，室温下将甲基三苯基溴化膦(1.3g，3.67mmol)溶解于四氢呋喃(20mL)中，在0℃下逐滴加入1M/L的叔丁醇钾四氢呋喃溶液(3.67mL，3.67mmol)，将混合物在室温下搅拌2小时。在室温下向反应液中逐滴加入(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-(苄基氧基)-4,4-二氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-酮121-1(400mg,0.73mmol)的四氢呋喃(5mL)溶液，将混合物在室温下搅拌3小时。TLC(石油醚/乙酸乙酯＝8/1，)监测反应。反应结束后，向反应液加入水(30mL)中，用乙酸乙酯(20mL×3)萃取，用饱和食盐水(40mL)洗涤萃取的有机溶液，用无水硫酸钠干燥，过滤，浓缩后得粗产品。粗产品经硅胶柱层析纯化分离(石油醚/乙酸乙酯＝8/1)，得到白色固体(6R)-6-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-(苄基氧基)-4,4-二氟-9a,11a-二甲基-6-甲亚基十六氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-醇121-2(158mg，纯度：90％，产率：35.68％)。1H 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).19F NMR(376MHz,CDCl3)δ＝-89.23(d,J＝238.5,1F),-110.87(d,J＝238.4,1F).
第三步：室温下50mL圆底烧瓶中，将(6R)-6-[(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-7-(苄基氧基)-4,4-二氟-9a,11a-二甲基-6-甲亚基十六氢-1H-环戊并[1,2-a]菲-1-基]-2-甲基庚-2-醇121-2(158mg,0.29mmol)溶解于四氢呋喃(2mL)和甲醇(2mL)，在室温下加入氢氧化锂，一水合物(73mg，1.74mmol)和水(1mL)，随后在室温下搅拌2小时，TLC(石油醚:乙酸乙酯＝5:1监测反应，反应结束后，用水(10mL)稀释，乙酸乙酯(10mL×3)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝5:1)，得到白色固体(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-4,4-二氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-6-甲亚基十六氢-1H-环戊并[1,2-a]菲-7-醇122(120mg，纯度:95％，产率：86.52％)。
化合物122：1H NMR(400MHz,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).13C NMR(101MHz,CDCl3)δ＝151.10,104.32,71.25,71.13,55.20,50.57,50.22,48.51,44.39,43.05,41.65,39.31,38.82,37.29,36.40,35.70,30.17,29.31,29.24,28.49,25.37,20.95,20.75,18.73,12.37,11.87.19F NMR(376MHz,CDCl3)δ＝-89.00(d,J＝237.7,1F),-110.95(d,J＝237.6,1F).LC-MS：[M+H-2H2O]+＝417.35.
第四步：在50mL圆底烧瓶中，室温下将(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-4,4-二氟-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-6-甲亚基十六氢-1H-环戊并[1,2-a]菲-7-醇122(213mg,0.47mmol)溶于四氢呋喃(12mL)，在0℃下逐滴加入二摩尔每升硼烷二甲硫醚络合物的四氢呋喃溶液(0.71mL，1.41mmol),将混合物在室温下搅拌3小时后,在0℃下向反应液中逐滴加入十摩尔每升氢氧化钠水溶液(0.28mL,2.82mmol)和百分之三十的过氧化氢水溶液(320mg,2.82mmol),将混合物在室温下搅拌16小时.TLC(石油醚/乙酸乙酯＝2/1)监测反应。反应结束后，向反应液加入水(20mL)中，用乙酸乙酯(20mL×3)萃取，用饱和食盐水(30mL)洗涤萃取的有机溶液，用无水硫酸钠干燥，过滤，浓缩后得粗产品。粗产品经硅胶柱层析纯化分离(洗脱剂：石油醚/乙酸乙酯＝2/1)，得到白色固体(1R,3aS,3bS,5aS,7S,9aS,9bS,11aR)-4,4-二氟-6-(羟基甲基)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-醇125(184mg，纯度：95％，产率：78.92％)。化合物125：1H NMR(400MHz,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).19F NMR(376MHz,CDCl3)δ＝-88.57(d,J＝237.7,1F),-110.59(d,J＝237.6,1F).13C 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,
第五步：室温下50mL圆底烧瓶中，将(1R,3aS,3bS,5aS,7S,9aS,9bS,11aR)-4,4-二氟-6-(羟基甲基)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-7-醇125(159mg,0.34mmol)溶解于1,2-二氯乙烷(8mL)，在室温下加入三乙胺(205mg，2.04mmol),4-二甲氨基吡啶(41mg,0.34mmol)和对甲基苯磺酰氯(129mg，0.68mmol)，随后在50℃下搅拌16小时，TLC(石油醚:乙酸乙酯＝3:1)监测反应。反应结束后，用水(20mL)淬灭，乙酸乙酯(20mL×3)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝3：1)，得到白色固体4-甲基苯磺酸-[(1R,3aS,3bS,5aS,7S,9aS,9bS,11aR)-4,4-二氟-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]甲基酯121-3(155mg，纯度:90％，产率：66.09％)。1H 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＝11.8,5.1,1H),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).
第六步：室温下50mL圆底烧瓶中，将4-甲基苯磺酸-[(1R,3aS,7S,9aS,11aR)-4,4-二氟-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]甲基酯121-3(45mg,0.072mmol)溶解于N,N-二甲基甲酰胺(2mL)，在室温下加入氰化钠(18mg，0.36mmol)，随后在100℃下 搅拌16小时，TLC(石油醚:乙酸乙酯＝4:1)监测反应。反应结束后，用水(10mL)淬灭，乙酸乙酯(10mL×3)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，得到白色固体[(1R,3aS,3bS,5aS,7S,9aS,9bS,11aR)-4,4-二氟-7-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-基]乙腈121(7.27mg，纯度:100％，产率：21.04％)。
化合物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).13C 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.
实施例123
化合物123：4-甲亚基-5α-胆甾-3β,25-二醇的制备
第一步：将反应物(5R)-5-[(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-乙酰氧基-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]己酸甲酯I-7(500mg,1.124 mmol,1.0eq)溶于四氢呋喃(40mL)中，将反应体系冷却至0℃后，缓慢加入甲基氯化镁(3.75mL,11.24mmol,3mol/L,10eq)，反应体系室温搅拌2h。TLC板(石油醚:乙酸乙酯＝5:1)监测反应。原料消耗完毕，停止反应。将20mL水加入到反应体系中，使用乙酸乙酯(50mL×3)萃取，收集有机相，使用无水硫酸钠干燥，有机相真空旋干得到粗品。粗品溶于乙酸乙酯中通过柱层析(石油醚:乙酸乙酯＝80：20)得到胆甾-6(5)-烯-3β,4β,19,25-四醇123-1(450mg，纯度:90％，收率:89.45％)。1H NMR(400MHz,CDCl3)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)。
第二步：胆甾-6(5)-烯-3β,4β,19,25-四醇123-1(450mg,1.118mmol,1.0eq)溶解在二氯甲烷(20mL)中,依次加入乙酸酐(0.315mL,3.354mmol,3.0eq)，4-二甲氨基吡啶(27.32mg,0.224mmol,0.2eq)和三乙胺(0.777mL,5.590mmol，5.0eq)，室温搅拌3小时，TLC(石油醚:乙酸乙酯＝5:1)监测反应。反应结束后，反应液用甲醇淬灭，反应液用饱和碳酸氢钠(10mL)和水(10mL)各洗涤一次，无水硫酸钠干燥，浓缩，在浓缩至干。柱层析(石油醚:乙酸乙酯＝90：10)得到白色固体乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯123-2(450mg,0.911mmol,纯度：90％，收率：81.49％)。1H NMR(400MHz,CDCl3)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)。
第三步：将乙酸-(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯123-2(450mg,1.012mmol,1.0eq)溶解在氯仿(20mL)中，氮气保护，加入二氧化硒(281mg,2.53mmol，2.5eq)和N-甲基吗啉(307.07mg,3.036mmol)，反应加热到75度搅拌48小时，TLC(石油醚:乙酸乙酯＝3:1)监测反应，原料基本消失，反应降至室温，加水淬灭，二氯甲烷萃取3次，合并有机相，无水硫酸钠干燥，过滤，滤液浓缩干，粗品快速柱层析纯化(石油醚:乙酸乙酯＝70：30)得到白色固体乙酸-(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯123-3(310mg,0.606mmol,纯度：90％，收率：59.85％)。1H NMR(400MHz,CDCl3)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)。
第四步：将化合物乙酸-(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a- 二甲基-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯123-3(40mg,0.087mmol，1.0eq)溶解在乙酸乙酯(25mL)中，然后加入二氧化铂(30mg,0.13mmol，1.5eq)并将反应体系置换成氢气氛围，将反应体系置于室温搅拌24h。TLC(石油醚:乙酸乙酯＝3:1)监测反应进度，原料消耗完毕。过滤反应体系，收集有机相并浓缩，得到粗品。粗品通过硅胶柱层析(石油醚:乙酸乙酯＝82:18to 81：19)得到白色固体乙酸-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯123-4(35mg,纯度90％，收率78.4％)。1H NMR(400MHz,CDCl3)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)。
第五步：将化合物乙酸-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-6-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯123-4(25mg,0.054mmol)溶解在二氯甲烷(5mL)中，氮气保护下，依次加入四丙基高钌酸铵(5.69mg,0.016mmol)，N-甲基吗啉氧化物(9.49mg,0.081mmol)和4A分子筛，室温搅拌，TLC(石油醚:乙酸乙酯＝3:1)监测反应。将水(10mL)加入到反应体系中，水层用EA(3×25mL)萃取。合并乙酸乙酯层并且用饱和食盐水(3×10mL)洗涤。乙酸乙酯层层用Na2SO4干燥，过滤和浓缩得到粗品。反应结束，反应液过滤，滤液水洗，有机相干燥，浓缩，粗品快速柱层析纯化(石油醚:乙酸乙酯＝81:19)得到白色固体乙酸-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-6-氧亚基十六氢-1H-环戊并[1,2-i]菲-7-基酯123-5(25mg,纯度90％,收率90％)。1H NMR(400MHz,CDCl3)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＝18.1,14.5Hz,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).
第六步：在50mL圆底烧瓶中，室温下将甲基三苯基溴化膦(4.65g,13.02mmol)悬浮于四氢呋喃(50mL)，在0℃下逐滴加入1M/L的叔丁醇钾四氢呋喃溶液(13.02mL,13.02mmol)，将混合物在室温下搅拌2小时后，在室温下向反应液中逐滴加入乙酸-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-6-氧亚基十六氢-1H-环戊并[1,2-i]菲-7-基酯123-5(1.2g,2.60mmol)的四氢呋喃(10mL)溶液，将混合物在室温下搅拌3小时。TLC(石油醚/乙酸乙酯＝10/1)监测反应。反应结束后，向反应液加入水(50mL)中，用乙酸乙酯(40mL×3)萃取，用饱和食盐水(80mL)洗涤萃取的有机溶液，用无水硫酸钠干燥，过滤，浓缩后得粗产品。粗产品经硅胶柱层析纯化分离(洗脱剂：石油醚/乙酸乙酯＝10/1)，得到白色固体乙酸-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-6-甲亚基十六氢-1H-环戊并[1,2-i]菲-7-基酯123-6(612mg，纯度：95％，产率：48.66％)。1H 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).
第七步：室温下50mL圆底烧瓶中，将乙酸-(1R,3aS,3bS,5aR,7S,9aR,9bS,11aR)-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基-6-甲亚基十六氢-1H-环戊并[1,2-i]菲-7-基酯123-6(700g,1.53mmol)溶解于四氢呋喃(10mL)和甲醇(10mL)，在室温下加入氢氧化锂，一水合物(385mg，9.16mmol)和水(5mL)，随后在室温下搅拌1小时，TLC(石油醚:乙酸乙酯＝4:1)监测反应。反应结束后，用水(30mL)稀释，乙酸乙酯(30mL×3)萃取，有机相饱和盐水(60mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝4:1)，得到白色固体4-甲亚基-5α-胆甾-3β,25-二醇123(614mg，纯度:97％，产率：93.67％)。
化合物123：1H NMR(400MHz,CDCl3)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).13C 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-2H2O]+＝381.3.
实施例137：
化合物137：24-[羟基(2-甲氧基苯基)甲基]-7-甲基-5α-胆烷-3β,4β-二醇的制备
第一步：将(9R)-9-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-11-甲亚基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳 -8-基]-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸烷106-5(25mg,0.038mmol)溶于甲醇(5mL)，加入Pd/C(13mg,0.061mmol)，将体系置换成氢气，混合液在40℃下搅拌1小时。经硅藻土过夜滤，旋干，得到粗品137-1(20mg)，直接投下一步。
第二步：将(9R)-9-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a,11-五甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸烷137-1(20mg,0.030mmol)溶于四氢呋喃(2mL)，加入盐酸(1mL,3.000mmol)，混合液在40℃下反应2小时。向反应液加入水，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，旋干。粗品经硅胶柱层析(石油醚/乙酸乙酯＝1/1)纯化得到24-[羟基(2-甲氧基苯基)甲基]-7-甲基-5α-胆烷-3β,4β-二醇白色固体137(4.5mg,0.007mmol,17.32％，两步的产率)。
化合物137：1H NMR(400MHz,CDCl3)δ7.32–7.27(m,1H),7.25–7.21(m,1H),6.95(t,J＝7.4Hz,1H),6.88(d,J＝8.2Hz,1H),4.85(dt,J＝9.6,5.8Hz,1H),3.86(s,3H),3.71(d,J＝2.2Hz,1H),3.68–3.64(m,0H),3.60–3.51(m,1H),1.96(d,J＝13.0Hz,1H),1.74(dd,J＝15.3,5.3Hz,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+H-H2O]+＝477.50.
实施例138：
化合物138：(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4,4-二氟-7-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-酮的制备

第一步：将化合物(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-7-(苄基氧基)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4,4-二氟-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-醇94-7(110mg,0.189mmol,1eq)溶于二氯甲烷(5mL)中，缓慢加入戴斯-马丁氧化剂(160mg,0.378mmol,2eq)，室温搅拌3小时,TLC(石油醚:乙酸乙酯＝4：1)监测。反应完成后向其中加入水摧灭,然后用乙酸乙酯(50mL)萃取,有机相用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,过滤,旋干得到粗品,粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝100:0to 3:1)得到(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-(苄基氧基)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4,4-二氟-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-酮白色固体138-1(109mg,纯度：97％,收率：73％)。1H NMR(400MHz,CDCl3)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).
第二步：室温下，将(1R,3aS,3bS,7S,9aR,9bS,11aR)-7-(苄基氧基)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4,4-二氟-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-酮138-1(15mg,0.025mmol，1.0eq)溶解于四氢呋喃(0.5mL)和甲醇(0.5mL)，在室温下加入氢氧化锂一水合物(11mg，0.25mmol，10.0eq)和水(0.2mL)，随后在室温下搅拌30min，TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，用水(10mL)稀释，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝100:0to 3:1)得到(1R,3aS,3bS,7S,9aR,9bS,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4,4-二氟-7-羟基-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-6-酮138(6.2mg，纯度100％,收率52％)为白色固体。
化合物138：1H NMR(400MHz,CDCl3)δ4.15(t,J＝8.2Hz,1H),3.49(d,J＝3.4Hz,1H),2.55–2.42(m,2H),2.11(d,J＝8.9Hz,1H),2.02(d,J＝12.5Hz,2H),1.91(d,J＝13.0Hz,2H),1.84(d,J＝5.5Hz,1H),1.73(s,2H),1.67(s,1H),1.62(d,J＝11.8Hz,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).19F NMR(377MHz,CDCl3)δ-90.22,-90.85,-111.49,-112.12,-115.16,-115.81,-116.09,-116.74.LC-MS:[M-H]+＝489.35。
实施例139：
化合物139：(1R,3aR,7S,9aR,9bR,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4-氟-9a,11a-二甲基- 2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6,7-二醇的制备
第一步：室温下，将(3S，4R，5R，8S，9S，10R，13R，14S，17R)-17-((R)-6-乙酰氧基-5,5-二氟-6-甲基庚烷-2-基)-10,13-二甲基-7-氧代十六氢-1H-环戊二烯[a]菲-3,4-二基二乙酸酯94-4(53mg,0.089mmol)溶解于二乙胺基三氟化硫(2mL)，在70℃下搅拌2小时，TLC(石油醚:乙酸乙酯＝4:1)监测反应，反应结束后，将反应液乙酸乙酯稀释，并慢慢滴加到冰水(20mL)中淬灭，乙酸乙酯(15mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)，得到白色固体乙酸-(6R)-6-[(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-6,7-二乙酰氧基-4-氟-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-3,3-二氟-2-甲基庚-2-基酯139-1(4.5mg，纯度:90％，产率：8.2％)。
第二步：室温下，将得到白色固体乙酸-(6R)-6-[(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-6,7-二乙酰氧基-4-氟-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-1-基]-3,3-二氟-2-甲基庚-2-基酯139-1(45mg,0.073mmol，1.0eq)溶解于四氢呋喃(1mL)和甲醇(1mL)，在室温下加入氢氧化锂一水合物(15mg，0.365mmol，5.0eq)和水(0.5mL)，随后在室温下搅拌30min，TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，用水(10mL)稀释，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝2:1)， 得到白色固体(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4-氟-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6,7-二醇139-2(28mg，纯度:90％，产率：77.78％)。
第三步：室温下，将(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4-氟-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6,7-二醇139-2(28mg,0.057mmol，1.0eq)溶解于二氯甲烷(2mL)和四氢呋喃(1mL)，在室温下加入三乙胺(35mg，0.34mmol，6.0eq)，4-二甲氨基吡啶(7mg,0.057mmol，1.0eq)和苯甲酰氯(24mg，0.17mmol，3.0eq)，随后在室温下搅拌8小时，TLC(石油醚:乙酸乙酯＝1:1)监测反应，反应结束后，用水(10mL)淬灭，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)和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:37degree)拆分，得到白色固体(苯甲酸-(1R,3aR,5aR,6R,7S,9aR,9bR,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4-氟-6-羟基-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-i]菲-7-基酯139-3(15mg，纯度:95％，产率：44.12％，保留时间tR＝1.624min)。1H NMR(400MHz,CDCl3)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＝13.4Hz,2H),1.14(s,3H),1.02(t,J＝10.2Hz,1H),0.95(d,J＝6.5Hz,3H),0.69(s,3H).
第四步：室温下，将(1R,3aR,7S,9aR,9bR,11aR)-7-(苄基氧基)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4-氟-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6-醇139-3(15mg,0.025mmol，1.0eq)溶解于四氢呋喃(0.5mL)和甲醇(0.5mL)，在室温下加入氢氧化锂一水合物(11mg，0.25mmol，10.0eq)和水(0.2mL)，随后在室温下搅拌30min，TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，用水(10mL)稀释，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，粗产物用快速色谱法分离纯化(石油醚:乙酸乙酯＝100:0to 3:1)得到(1R,3aR,7S,9aR,9bR,11aR)-1-[(2R)-5,5-二氟-6-羟基-6-甲基庚-2-基]-4-氟-9a,11a-二甲基-2,3,3a,5,5a,6,7,8,9,9a,9b,10,11,11a-十四氢-1H-环戊并[1,2-a]菲-6,7-二醇白色固体139(10mg，纯度100％,收率79％)。
化合物139：1H NMR(400MHz,CDCl3)δ3.85(s,1H),3.60(d,J＝11.0Hz,1H),2.66(t,J＝12.4Hz,1H),2.14(s,1H),2.01(t,J＝11.5Hz,3H),1.89(s,2H),1.85(d,J＝4.1Hz,1H),1.77(s,2H),1.71(s,1H),1.63–1.55(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).19F NMR(377MHz,CDCl3)δ-109.66,-115.15,-115.80,-116.12,-116.77.LC-MS:[M-H]+＝471.35。
实施例143：
化合物143：7-氨基-24-[羟基(2-甲氧基苯基)甲基]-5α-胆烷-3β,4β-二醇的制备
第一步：将化合物(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-酮106-4(50mg,0.075mmol，1.0eq)溶解在四氢呋喃(2mL)和甲醇(1mL)的混合溶剂中,加入催化量乙酸(0.001mL,0.014mmol),乙酸铵(57.81mg,0.750mmol，10eq)，所得混合液在反应液在N2，25℃中搅拌1h，随后加入氰基硼氢化钠[还原剂](47.13mg,0.750mmol，10eq)，继续搅拌2小时，通过TLC板(二氯甲烷:甲醇＝10:1)监测反应进度。加水淬灭，水层二氯甲烷(3×10mL)萃取。合并二氯甲烷层并且用饱和食盐水(5mL)洗涤，无水Na2SO4干燥，过滤和浓缩得到粗品,粗品硅胶柱层析纯化(二氯甲烷:甲醇＝10:1)得到白色固体(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-胺143-1(30mg,0.043mmol,纯度：90％，收率：56.87％)。LC-MS：[M+H]+＝668.4。
第二步：将化合物(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-胺143-1(30mg,0.05mmol,1.0eq)溶解在四氢呋喃(5mL)中,加入稀盐酸(0.05mL)，25℃中搅拌1小时，继续搅拌2小时，通过TLC(二氯甲烷:甲醇＝5:1)监测反应进度。加水淬灭，水层二氯甲烷(3×10mL)萃取。合并二氯甲烷层并且用饱和食盐水(5mL)洗涤，无水Na2SO4干燥，过滤和浓缩得到粗品,粗品硅胶柱层析纯化(二氯甲烷:甲醇＝10:1)得到白色 固体7-氨基-24-[羟基(2-甲氧基苯基)甲基]-5α-胆烷-3β,4β-二醇143(17.79mg,0.034mmol,纯度:97.15％,收率:74.92％)。
化合物143：1H NMR(400MHz,DMSO)δ7.55(s,1H),7.38(m,1H),7.18(t,J＝7.8Hz,1H),6.92(m,2H),4.86(s,2H),4.42(d,J＝5.8Hz,1H),4.17(t,J＝6.3Hz,1H),3.75(d,J＝3.0Hz,3H),3.44(d,J＝22.4Hz,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。
实施例144：
化合物144：4-甲基-5α-胆甾-3β,25-二醇的制备
第一步：室温下，将(3S，5R，8S，9S，10R，13R，14S，17R)-17-((R)-6-羟基-6-甲基庚烷-2-基)-10,13-二甲基-4-亚甲基六氢-1H-环戊并[a]菲-3-醇123(25mg，0.060mmol,1eq)溶于二氯甲烷(2mL),加入DMAP(2mg，0.016mmol,0.3eq)和三乙胺(36mg，0.36mmol,6eq)，在冰浴下冷却至0℃，慢慢滴加乙酸酐(12mg，0.12mmol,2eq)的二氯甲烷(0.5mL)溶液。10min后升到室温并搅拌6小时。反应液用水淬灭，用二氯甲烷萃取(10mL x 2)。有机相合并后用饱和食盐水洗(20mL),经无水硫酸钠干燥，过滤，浓缩后得到粗品。经层析柱分离(4g,0～10％EtOAc/PE，20mL/min)得到(R)-6-((3S，5R，8S，9S，10R，13R，14S，17R)-3-乙酰氧基-10,13-二甲基-4-亚甲基杂十氢-1H-环戊基菲17-基)-2-甲基庚-2-基乙酸酯144-1(10mg,纯度：90％,产率43.3％)。1H 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).
第二步：室温下，将(R)-6-((3S，5R，8S，9S，10R，13R，14S，17R)-3-乙酰氧基-10,13-二甲基-4-亚甲基杂十氢-1H-环戊基菲17-基)-2-甲基庚-2-基乙酸酯144-1(10mg,0.020mmol,1eq)溶解于 乙酸乙酯(3mL)，加入10％Pd/C(20mg)和乙酸(0.1mL)，室温下将混合物在氢气氛围中搅拌16小时。HNMR检测反应。反应结束后，用乙酸乙酯(10mL)稀释反应液，通过硅藻土过滤，滤液依次用饱和碳酸氢钠溶液(10mL)和饱和食盐水洗(10mL),经无水硫酸钠干燥，过滤，浓缩后得到白色固体(6R)-6-((3S，5S，8S，9S，10R，13R，14S，17R)-3-乙酰氧基-4,10,13-三甲基十六氢-1H-环戊二烯菲-17-基)-2-甲基庚烷-2-基乙酸酯144-2(5mg，纯度：90％，产率：44.8％)。1H NMR(400MHz,CDCl3)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).
第三步:室温下，将(6R)-6-((3S，5S，8S，9S，10R，13R，14S，17R)-3-乙酰氧基-4,10,13-三甲基十六氢-1H-环戊二烯菲-17-基)-2-甲基庚烷-2-基乙酸酯144-2(5mg,0.010mmol，1.0eq)溶解于四氢呋喃(0.5mL)和甲醇(0.5mL)，在室温下加入氢氧化锂一水合物(8mg，0.19mmol，19.2eq)和水(0.2mL)，随后在室温下搅拌2小时和60℃油浴中搅拌3小时，通过TLC板(石油醚:乙酸乙酯＝4:1，磷钼酸烤板)检测反应完全。反应结束后，用水(10mL)稀释，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10:1)，得到白色固体4-甲基-5α-胆甾-3β,25-二醇144(3.07mg，纯度:99.6％，产率：38.3％)。
化合物144:1H NMR(400MHz,CDCl3)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).
实施例158&159：
化合物158[(1R,3aS,3bS,4Z,6R,7S,9aR,9bS,11aR)-6,7-二羟基-1-[(2R)-6-羟基-6-(2-甲氧基苯基)己-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-4-亚基]乙腈和化合物159[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二羟基-1-[(2R)-6-羟基-6-(2-甲氧基苯基)己-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-4-基]乙腈的制备。

第一步：冰水浴下，将正丁基锂(0.180mL,0.450mmol)溶液滴加到氰甲基磷酸二乙酯(87.63mg,0.495mmol)溶在四氢呋喃(5mL)的溶液中，搅拌30分钟后，将(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-酮106-4(30mg,0.045mmol)溶在四氢呋喃中后滴加到反应瓶中，混合液在室温下搅拌2小时。反应液用饱和氯化铵溶液猝灭，加乙酸乙酯萃取，有机相用无水硫酸钠干燥，旋干得到粗品。粗品经快速色谱仪(石油醚/乙酸乙酯＝8％)纯化得到[(3aS,5aR,5bS,7aR,8R,10aS,10bS,11Z,12aR,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-11-亚基]乙腈无色透明固体159-1(45mg,0.055mmol,61.62％)。1H NMR(400MHz,CDCl3)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).
第二步：将[(3aS,5aR,5bS,7aR,8R,10aS,10bS,11Z,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-11-亚基]乙腈159-1(40mg,0.058mmol)溶于甲醇(5mL)中，加入Pd/C(20mg,0.094mmol)，置换成氢气，混合液在40℃下搅拌2小时。TLC(石油醚/乙酸乙酯＝5/1)追踪反应液发现原料反应完，有极性变大的新点。经硅藻土过滤Pd/C，旋干。粗品经快速色谱仪(石油醚/乙酸乙酯＝10％)纯化得到[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-11-基]乙腈159-2白色固体(20mg,0.025mmol,42.35％)。1H 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＝15.0Hz,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).
第三步：将[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-11-基]乙腈159-2(35mg,0.051mmol)溶于四氢呋喃(3mL)中，加入盐酸(2mL,6.000mmol)，混合液在室温下搅拌2小时。向反应液加水，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，旋干。粗品经快速色谱仪(石油醚/乙酸乙酯＝50％)纯化得到[(1R,3aS,3bS,6R,7S,9aR,9bS,11aR)-6,7-二羟基-1-[(2R)-6-羟基-6-(2-甲氧基苯基)己-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-4-基]乙腈159(5mg,0.008mmol,15.63％)为白色固体。
化合物159：1H NMR(400MHz,CDCl3)δ7.31–7.27(m,1H),7.26–7.21(m,1H),6.96(t,J＝7.4Hz,1H),6.88(d,J＝8.2Hz,1H),4.89–4.80(m,1H),3.86(s,3H),3.74(s,1H),3.56(d,J＝10.7Hz,1H),2.54(d,J＝4.5Hz,2H),1.99(d,J＝12.1Hz,2H),1.86(t,J＝10.0Hz,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+H-H2O]+＝520.45,tR＝6.941min.
第四步：将[(3aS,5aR,5bS,7aR,8R,10aS,10bS,11Z,12bR)-8-[(9R)-5-(2-甲氧基苯基)-2,2,3,3-四甲基-4-氧杂-3-硅杂癸-9-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-11-亚基]乙腈159-1(40mg,0.058mmol)溶于四氢呋喃(3mL)中，加入盐酸(2mL,6.000mmol)，混合液在室温下搅拌2小时。向反应液加水，用乙酸乙酯萃取,有机相无水硫酸钠干燥。粗品经快速色谱仪(石油醚/乙酸乙酯＝50％)纯化得到 [(1R,3aS,3bS,4Z,6R,7S,9aR,9bS,11aR)-6,7-二羟基-1-[(2R)-6-羟基-6-(2-甲氧基苯基)己-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-a]菲-4-亚基]乙腈158(15mg,0.024mmol,42.03％)为白色固体。
化合物158：1H NMR(400MHz,CDCl3)δ7.33–7.27(m,1H),7.26–7.20(m,1H),6.96(t,J＝7.5Hz,1H),6.89(d,J＝8.2Hz,1H),5.06(s,1H),4.85(dt,J＝9.6,5.8Hz,1H),3.86(s,4H),3.62–3.52(m,1H),2.73(dd,J＝12.7,2.3Hz,1H),2.55(t,J＝13.0Hz,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).13C NMR(101MHz,CDCl3)δ156.57,132.56,128.27,127.01,126.88,120.76,117.66,110.57,90.13,74.06,71.84,71.35,70.95,56.44,55.28,54.96,51.72,49.47,43.80,42.94,38.33,37.74,37.55,36.48,36.28,35.99,35.79,35.61,35.47,35.40,31.54,28.06,25.64,25.07,22.70,22.56,18.74,12.13,-0.01.LCMS(ESI)[M+H-2H2O]+＝500.37,tR＝5.860min.
实施例160：
化合物160 25-羟基-4β-羟基-3β-羟基-5α-胆甾-7-甲腈的制备。

第一步：室温下，将(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-2,2,5a,7a-四甲基-11-氧亚基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己酸甲酯I-12(200mg,0.421mmol，1eq)溶解于甲醇(5mL)，氮气保护下，降温到0℃，加入硼氢化钠(24mg，0.632mmol，1.5eq)，随后转至室温搅拌2小时，TLC(石油醚:乙酸乙酯＝2：1)监测反应，反应结束后，用水(30mL)淬灭，乙酸乙酯(30mL×2)萃取，有机相饱和盐水(50mL)洗涤，无水硫酸钠干燥，旋干有机相得到白色固体(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11-羟基-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己酸甲酯160-1(200mg，纯度:95％，产率：99％)，直接往后投。
第二步：室温下，将(5R)-5-[(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-11-羟基-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':1,2]菲并[7,8-d][1,3]二氧杂环戊熳-8-基]己酸甲酯160-1(200mg,0.42mmol)溶解于无水四氢呋喃(5mL)，氮气保护下，降温到0℃，滴加甲基溴化镁(3M in 2-Me-THF，0.86mL，4.19mmol，10eq)，随后转至室温搅拌2小时，TLC(石油醚:乙酸乙酯＝2：1)监测反应，反应结束后，体系降温至0℃，用饱和氯化铵水溶液(30mL)淬灭，乙酸乙酯(30mL×2)萃取，有机相饱和盐水(50mL)洗涤，无水硫酸钠干燥，硅胶柱层析纯化(石油醚:乙酸乙酯＝4：1)，得到白色固体(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(2R)-6-羟基-6-甲基庚-2-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-醇160-2(139mg，纯度:95％，产率：67％)。1H NMR(400MHz,CDCl3)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).
第三步：室温下将(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(2R)-6-羟基-6-甲基庚-2-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-醇160-2(130mg,0.273mmol，1.0eq)溶解于四氢呋喃(10mL)，在室温下加入戴斯马丁氧化剂(424mg，0.545mmol,2eq)，随后在室温下搅拌1小时，TLC(石油醚:乙酸乙酯＝2:1)监测反应，反应结束后，用冰水(50mL)淬灭，乙酸乙酯(30mL×2)萃取，有机相饱和盐水(50mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝5：1)，得到白色固体(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(2R)-6-羟基-6-甲基庚-2-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-酮160-3(110mg，纯度:95％，产率：85％)。1H NMR(400MHz,CDCl3)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＝6.8,4.5Hz,2H),1.21(s,6H),1.13–0.99(m,5H),0.92(d,J＝6.4Hz,3H),0.66(s,3H).
第四步：室温下，将叔丁醇钾(106mg,1.95mmol，1.0eq)溶解于二甲基甲酰胺(3mL)，在室温下加入对甲基苯磺酰甲基异腈(79mg，0.38mmol，1.5eq)，随后在室温下搅拌5分钟后加入甲醇(8.1mg，0.25mmol，1eq)，室温下搅拌10分钟后加入(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(2R)-6-羟基-6-甲基庚-2-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-酮160-3(120mg,0.253mmol，1.0eq)，室温下搅拌12小时，TLC(石油醚:乙酸乙酯＝3:1)监测反应，反应结束后，用水(30mL)淬灭，乙酸乙酯(20mL×2)萃取，有机相饱和盐水(30mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝15：1)，得到白色固体(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(2R)-6-羟基-6-甲基庚-2-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-甲腈160-4(30mg，纯度:90％，产率：25％)。化合物160-4：1H NMR(400MHz,CDCl3)δ4.05(dt,J＝8.4,5.1Hz,2H),2.93(dd,J＝6.6,4.4Hz,1H),2.00–1.95(m,2H),1.82–1.60(m,10H),1.50(s,3H),1.38(ddd,J＝15.5,11.4,5.4Hz,10H),1.30(s,3H),1.25(s,5H),1.22(s,6H),1.05(s,3H),0.93(t,J＝6.0Hz,3H),0.75–0.62(m,3H).
第五步：室温下，将(3aS,5aR,5bS,7aR,8R,10aS,10bS,12aR,12bR)-8-[(2R)-6-羟基-6-甲基庚-2-基]-2,2,5a,7a-四甲基-4,5,5a,5b,6,7,7a,8,9,10,10a,10b,11,12,12a,12b-十六氢-3aH-环戊并[1',2':7,8]菲并[1,2-d][1,3]二氧杂环戊熳-11-甲腈160-4(30mg，0.06mmol，1eq)溶解于四氢呋喃(3mL)和3M盐酸(1ml)，随后在室温下搅拌30min，TLC(石油醚:乙酸乙酯＝1:1)监测反应，反应结束后，用水(10mL)稀释，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩 有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝2:1)，得到白色固体25-羟基-4β-羟基-3β-羟基-5α-胆甾-7-甲腈160-5(15mg，纯度:80％，产率：50％)。1H NMR(400MHz,CDCl3)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),1.25(s,3H),1.22(s,3H),1.04(s,3H),0.92(d,J＝6.5Hz,3H),0.66(s,3H).
第六步：室温下，将25-羟基-4β-羟基-3β-羟基-5α-胆甾-7-甲腈160-6(10mg,0.022mmol，1.0eq)溶解于二氯甲烷(2mL)，在室温下加入三乙胺(7mg，0.07mmol，3.0eq)，4-二甲氨基吡啶(2.2mg,0.018mmol，0.8eq)和苯甲酰氯(5mg，0.034mmol，1.5eq)，随后在室温下搅拌1小时，TLC(石油醚:乙酸乙酯＝1:1)监测反应，反应结束后，用水(10mL)淬灭，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝10：1)和手性拆分(仪器:Waters Acquity UPCC，色谱柱:Daicel CHIRALPAK IB 4.6*250mm,5um，流动相:CO2/MeOH(0.1％DEA)＝60/40，流速:1.5ml/min，柱温:37degree)拆分得到白色固体苯甲酸-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4-氰基-6-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯160-6(8mg，纯度:95％，产率：80％，保留时间：1.595min)。1H NMR(400MHz,CDCl3)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).
第七步：室温下，将白色固体苯甲酸-(1R,3aS,3bS,5aR,6R,7S,9aR,9bS,11aR)-4-氰基-6-羟基-1-[(2R)-6-羟基-6-甲基庚-2-基]-9a,11a-二甲基十六氢-1H-环戊并[1,2-i]菲-7-基酯160-6(20mg，0.045mmol，1eq)溶解于四氢呋喃(0.5mL)和甲醇(0.5mL)，在室温下加入氢氧化锂一水合物(11mg，0.25mmol，10.0eq)和水(0.2mL)，随后在室温下搅拌30min，TLC(石油醚:乙酸乙酯＝10:1)监测反应，反应结束后，用水(10mL)稀释，乙酸乙酯(10mL×2)萃取，有机相饱和盐水(20mL)洗涤，无水硫酸钠干燥，浓缩有机相，硅胶柱层析纯化(石油醚:乙酸乙酯＝3:1)，得到白色固体25-羟基-4β-羟基-3β-羟基-5α-胆甾-7-甲腈160(12mg，纯度:98.73％，产率：60％)。
化合物160：1H NMR(400MHz,CDCl3)δ3.76(s,1H),3.66(s,1H),2.93(dd,J＝4.5,2.3Hz,1H),2.28(dd,J＝37.9,7.5Hz,2H),2.16–2.04(m,2H),2.04–1.94(m,3H),1.89(dd,J＝8.1,4.5Hz,1H),1.73(dd,J＝8.7,4.9Hz,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：[M-H]+＝445.36.
效果实施例1本发明化合物对SREBP通路抑制效果
1.1：SREBP荧光素酶报告基因系统
将Huh-7/SRE-luc细胞孵育于固醇缺乏的培养基中(5％去脂蛋白血清，2μM洛伐他汀，10μM甲羟戊酸)同时添加相应浓度的化合物处理16小时。其中，Huh-7/SRE-luc细胞为人肝癌细胞系Huh-7中稳定表达LDLR promotor-luciferase和绿荧光蛋白(GFP)的细胞。该细胞由Huh-7转染pLDLR-promotor-luciferase、pEGFP-N1质粒，并经过G418抗生素选择压力筛选后获得。其中Huh-7细胞购自ATCC，pLDLR-promotor-luciferase质粒经分子克隆方法获得，pEGFP-N1质粒购自Addgene。
化合物处理结束后，细胞用裂解液(E397A，Promega)裂解，添加荧光素酶底物(E1500，Promega)后，SRE驱动的荧光素酶的活性通过BioTek Synergy HTX酶标仪(包含但不限于此一类的仪器)测量。绿色荧光蛋白(EGFP)的荧光强度也是通过上述的BioTek酶标仪(包含但不限于此一类的仪器)测量，并作为内参。SRE驱动的荧光素活性除以绿色荧光蛋白的荧光强度后得到的比值作为SREBP通路活性的指标。每一个受试化合物的测试数据经prism软件分析(以DMSO处理组为参照计算出化合物抑制率，使用Prism log(inhibitor)VS.response--Variable slope(four-parameter)模型进行拟合,计算公式：Y＝Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope)))，得到该化合物的IC50参数。
1.2本发明化合物对SREBP通路抑制效果
通过生物测试实施例1.1的方法测试本发明化合物对SREBP通路的抑制效果，每个化合物浓度梯度设计为0.01、0.03、0.1、0.3、1.0、3.0、10μM，对照为溶媒DMSO。部分化合物的IC50值如表1中所示。
表1








效果实施例2：小鼠肝微粒体代谢稳定性实验
分别配制PB溶液(100mM)、MgCl2溶液(300mM)、NADPH溶液(2.04mM)和小鼠肝微粒体(SHQY货号：M1000批号：2110330)溶液(1.00mg/mL)，然后分别用二甲基亚砜(DMSO)配制化合物和睾酮(阳性对照，半衰期为3.1分钟)储备液，用甲醇稀释至100μM用于样品孵育，-10～-30℃保存。然后将100μL的小鼠肝微粒体溶液加入到96孔样品板中。对于0min的样品，首先加入含内标的乙腈溶液；其次加入2μL 100μM的工作液；然后加入98μL已预热到37℃的2.04mM NADPH。对于5min、15min、30min、45min和60min的样品，首先加入2μL 100μM的工作液；其次加入98μL已预热到37℃的2.04mM NADPH启动反应；到达相应的时间点后，加入含内标的乙腈溶液终止反应。对于NCF60的样品，首先加入2μL 100μM的工作液；其次加入98μL已预热到37℃的PB启动反应；到达相应的时间点后，加入含内标的乙腈溶液终止反应。实验过程中，样品均在37℃水浴锅中孵育。将所有样品在振荡器上以800转/分振荡10min，然后在3200g离心20min。吸取上清液，与去离子水以相应的比例进行稀释后用LC-MS/MS进样分析。采用化合物在5、15、30、45和60min的峰面积比除以0min的峰面积比得到每个时间点化合物的剩余率，然后用Excel计算化合物的半衰期(T1/2)。
表2：部分实施例的化合物的小鼠肝脏微粒体稳定性结果

效果实施例3：测试本发明化合物在禁食状态模拟肠液(FaSSIF)中的溶解度
用DMSO溶解化合物粉末，将其配制成10.0mM储备液待用。称量0.021g NaOH、0.198g NaH2PO4和0.309g NaCl，加入到50.0mL的超纯水中，混匀后用HCl或NaOH调节pH值至6.5，然后添加0.112g的FaSSIF粉末，待完全溶解后在室温静置2小时备用。取一定体积的化合物储备液加入到一定体积的FaSSIF中，配制成300μM的样品溶液，在室温以及1000rpm的条件下混匀1小时。1小时后，将样品离心并取一定体积的上清液，加入终止液，沉淀样品，混匀并离心20分钟。然后再取上清液并用稀释液进行稀释后用液相色谱-串联质谱(LC-MS/MS)方法分析。样品中待测化合物的浓度采用LC-MS/MS法进行定量测定。
表3：部分实施例的化合物的FASSIF溶解度结果
效果实施例4：化合物对FASN基因表达的抑制实验
将Huh-7细胞(购自ATCC)用DMEM培养基(含有10％胎牛血清LONSERA S711-001S)培养于37℃、5％CO2培养箱中培养。24小时后吸掉原有的培养基，换成DMEM饥饿培养基(含5％去脂蛋白血清(LPDS)，2μM洛伐他汀(-lovastatin),10μM甲羟戊酸(mevalonate))对照组加二甲亚砜(DMSO),化合物处理组加不同浓度的化合物。化合物最高浓度3000nM,3倍稀释，共5个浓度。培养16小时。进行细胞裂解，cell to cDNA逆转录，以及qPCR检测SREBP靶基因脂肪酸合成酶(Fatty AcidSynthase，FASN)基因的表达(FASN基因正向引物序列为：5’-AAGGACCTGTCTAGGTTTGATGC-3’，反向引物序列为：5’-TGGCTTCATAGGTGACTTCCA-3’)，计 算IC50。每一个受试化合物的测试数据经prism软件分析(以DMSO处理组为参照计算出化合物抑制率，使用Prism log(inhibitor)VS.response--Variable slope(four-parameter)模型进行拟合,计算公式为Y＝Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope)))，得到该化合物的IC50参数。
表4：部分实施例的化合物对FASN基因表达的抑制结果
本发明中，对比化合物1的结构为
虽然以上描述了本发明的具体实施方式，但是本领域的技术人员应当理解，这些仅是举例说明，在不背离本发明的原理和实质的前提下，可以对这些实施方式做出多种变更或修改。因此，本发明的保护范围由所附权利要求书限定。

Claims (10)

  1. 一种式I、II或III所示化合物或其药学上可接受的盐：
    其中，
    R4a为H；R4b为卤素、CN、-CH2-CN、-CH2-OH、-CH(CH3)-OH、-COOH、-CH2-COOH、C1-C6卤代烷基、-CH2-OAc或NH2；或者，R4a和R4b以及它们连接的碳原子一起共同形成
    当7号碳原子和8号碳原子之间为单键时，R7a和R7b独立地为H、卤素、CN、-CH2-OH、-COOH、-CH2-COOH、NH2、C1-C6烷基、C1-C6卤代烷基或-CH2-CN；或者，R7a和R7b以及它们连接的碳原子一起共同形成R8a为H；
    当7号碳原子和8号碳原子之间为双键时，R7a不存在，R7b为卤素；R8a不存在；
    R21为
    n1和n3独立地为2、3、4或5；
    m1和m3独立地为0、1、2、3、4或5；
    环A和环C独立地为C6-C10芳基、“杂原子选自N、O和S中的1种、2种或3种，杂原子数为 1个、2个或3个的5-10元杂芳基”或C3-C6环烷基；
    RA和RC独立地为卤素、C1-C6烷基、C1-C6卤代烷基、C1-C6烷氧基或C1-C6卤代烷氧基；
    R1为或NR1aR1b；
    R1a和R1b独立地为H或C1-C6烷基；
    R4c为H；R4d为H或OH；
    R7c为H，R7d独立地为CN、-CH2-OH、-COOH、C1-C6烷基、C1-C6卤代烷基、-CH2-COOH、NH2、-CH2-CN或Cl；或者，R7c和R7d以及它们连接的碳原子一起共同形成
    R22为
    n2为2、3、4或5；
    m2为0、1、2、3、4或5；
    环B为C6-C10芳基、“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个、2个或3个的5-10元杂芳基”或C3-C6环烷基；
    RB独立地为卤素、C1-C6烷基、C1-C6卤代烷基、C1-C6烷氧基或C1-C6卤代烷氧基；
    R3a为H或C1-C6烷基；
    R3b为-O-(CH2)n-R3-1、NR3-2R3-3或OH；当R3b为OH时，R3a为C1-C6烷基；
    n为1、2、3或4；R3-1为OH或-C(O)O-R3-1a；R3-1a为H或C1-C6烷基；
    R3-2和R3-3独立地为H或-S(O)2R3-2a；R3-2a为C1-C6卤代烷基或C1-C6烷基；
    R4e为H；R4f为H、OH或-O-(CH2)m-O-R4-1；或者R4e和R4f以及它们连接的碳原子一起共同形成m为1或2；R4-1为C1-C6烷基；
    带“*”碳原子表示当为手性碳原子时，独立地为R构型、S构型或者两者的混合；
    带“#”碳原子表示当为手性碳原子时，独立地为R构型、S构型或者两者的混合；
    带“&”碳原子表示当为手性碳原子时，独立地为R构型、S构型或者两者的混合；
    如式I所示化合物不为以下化合物：
    及其异构体；
    如式II所示化合物不为以下化合物：
    及其异构体；
    如式III所示化合物不为以下化合物：
    及其异构体。
  2. 如权利要求1所述的式I、II或III所示化合物或其药学上可接受的盐，其特征在于，满足下述条件中的一个或多个：
    (1)R4b中，所述卤素独立地为F、Cl、Br或I；例如Br；
    (2)R4b中，所述C1-C6卤代烷基独立地为CHF2、CH2F或CF3；
    (3)R7a和R7b中，所述卤素独立地为F、Cl、Br或I；优选为F；
    (4)R7a和R7b中，所述C1-C6烷基独立地为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基；
    (5)R7a和R7b中，所述C1-C6卤代烷基独立地为CHF2、CH2F或CF3，例如CHF2；
    (6)环A和环C中，所述C6-C10芳基独立地为苯基或萘基，优选为苯基；
    (7)环A和环C中，所述“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个、2个或3个的5-10元杂芳基”独立地为“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个或2个的5-6元或9-10杂芳基”，例如为吡啶基(例如)；
    (8)环A和环C中，所述C3-C6环烷基独立地为环丙基、环丁基、环戊基或环己基；
    (9)RA和RC中，所述卤素独立地为F、Cl、Br或I；优选为F或Cl，例如F；
    (10)RA和RC中，所述C1-C6烷基独立地为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基；
    (11)RA和RC中，所述C1-C6卤代烷基独立地为CHF2、CH2F或CF3；
    (12)RA和RC中，所述C1-C6烷氧基独立地为甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、异丁氧基、仲丁氧基或叔丁氧基，优选为甲氧基或乙氧基；
    (13)RA和RC中，所述C1-C6卤代烷氧基独立地为-OCHF2、-OCH2F或-OCF3；优选为-OCF3；
    (14)R1a和R1b中，所述C1-C6烷基独立地为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基，例如甲基；
    (15)式I中，带“*”碳原子表示当为手性碳原子时，为S构型；
    (16)R7c和R7d中，所述C1-C6烷基独立地为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基；
    (17)R7c和R7d中，所述C1-C6卤代烷基独立地为CHF2、CH2F或CF3，例如CHF2；
    (18)环B中，所述C6-C10芳基为苯基或萘基，优选为苯基；
    (19)环B中，所述“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个、2个或3个的5-10元杂芳基”为“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个或2个的5-6元或9-10杂芳基”，例如为吡啶基(例如)；
    (20)环B中，所述C3-C6环烷基为环丙基、环丁基、环戊基或环己基；
    (21)RB中，所述卤素独立地为F、Cl、Br或I；优选为F或Cl，例如F；
    (22)RB中，所述C1-C6烷基独立地为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基；
    (23)RB中，所述C1-C6卤代烷基独立地为CHF2、CH2F或CF3；
    (24)RB中，所述C1-C6烷氧基独立地为甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、异丁氧基、仲丁氧基或叔丁氧基，优选为甲氧基或乙氧基；
    (25)RB中，所述C1-C6卤代烷氧基独立地为-OCHF2、-OCH2F或-OCF3；优选为-OCF3；
    (26)式II中，带“*”碳原子表示当为手性碳原子时，为S构型；
    (27)式II中，带“#”碳原子表示当为手性碳原子时，为R构型；
    (28)R3a中，所述C1-C6烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基，例如甲基；
    (29)R3-1a中，所述C1-C6烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基，例如叔丁基；
    (30)R3-2a中，所述C1-C6卤代烷基为CHF2、CH2F或CF3，例如CF3；和
    (31)R4-1中，所述C1-C6烷基为甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基，例如乙基。
  3. 如权利要求1所述的式I、II或III所示化合物或其药学上可接受的盐，其特征在于，满足下述条件中的一个或多个：
    (1)式I中，为
    (2)R4a为H；R4b为卤素(例如Br)、CN、-CH2-CN、-CH2-OH、-CH(CH3)-OH、-COOH、-CH2-COOH、-CH2-OAc或NH2；或者，R4a和R4b以及它们连接的碳原子一起共同形成
    (3)当7号碳原子和8号碳原子之间为单键时，R7a和R7b独立地为H或卤素(例如F)，较佳的，R7a和R7b同时为H或卤素(例如F)；
    (4)当7号碳原子和8号碳原子之间为双键时，R7a不存在，R7b为卤素(例如F)；R8a不存在；
    (5)R21为
    (6)环A和环C独立地为C6-C10芳基或“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个、2个或3个的5-10元杂芳基”；
    (7)RA和RC独立地为卤素、C1-C6烷氧基或C1-C6卤代烷氧基；
    (8)n1为3；
    (9)n3为3；
    (10)m1为0、1、2或3；
    (11)m3为0、1、2或3；
    (12)式II中，为
    (13)R7c为H，R7d独立地为CN、-CH3、-CH2-OH、-COOH、-CHF2、-CH2-COOH、NH2、-CH2-CN或Cl；或者，R7c和R7d以及它们连接的碳原子一起共同形成
    (14)R22为
    (15)环B为C6-C10芳基或“杂原子选自N、O和S中的1种、2种或3种，杂原子数为1个、2个或3个的5-10元杂芳基”；
    (16)RB独立地为卤素、C1-C6烷氧基或C1-C6卤代烷氧基；
    (17)n2为3；
    (18)m2为0、1、2或3；
    (19)式III中，为
    (20)n为1、2或3；
    (21)R3-2为H，R3-3为-S(O)2R3-2a；和
    (22)m为1。
  4. 如权利要求1所述的式I、II或III所示化合物或其药学上可接受的盐，其特征在于，满足下述条件中的一个或多个：
    (1)R4a为H；R4b为Br、CN、-CH2-CN、-CH2-OH、-CH(CH3)-OH、-COOH、-CH2-OAc或NH2；或者，R4a和R4b以及它们连接的碳原子一起共同形成
    (2)当7号碳原子和8号碳原子之间为单键时，R7a和R7b同时为H或卤素(例如F)；
    (3)R7c为H，R7d独立地为CN、-CH2-OH、-CH3、NH2、-CH2-CN、-CF2H、-COOH，或者，R7c和R7d以及它们连接的碳原子一起共同形成
    (4)R3a为H，R3b为-NH2、 或者，R3a为甲基，R3b为OH；和
    (5)R4e为H；R4f为H、OH或或者R4e和R4f以及它们连接的碳原子一起共同形成
  5. 如权利要求1所述的式I、II或III所示化合物或其药学上可接受的盐，其特征在于，满足下述条件中的一个或多个：
    (1)为
    (2)为
    (3)式I中，为
    (4)为
    (5)式II中，为
    (6)式III中，为
  6. 如权利要求1所述的式I、II或III所示化合物或其药学上可接受的盐，其特征在于，满足下述条件中的一个或多个：
    (1)R21为
    (2)R22为
  7. 如权利要求1所述的式I、II或III所示化合物或其药学上可接受的盐，其特征在于，如下任一所示的化合物或其药学上可接受的盐：








  8. 一种药物组合物，其包含如权利要求1-7任一项所述的化合物或其药学上可接受的盐，以及至少一种药用辅料。
  9. 一种如权利要求1-7任一项所述的化合物或其药学上可接受的盐或如权利要求8所述的药物组合物在制备用于预防和/或治疗疾病的药物中的应用，其中所述疾病为肥胖、高脂血症、脂肪肝、糖尿病、动脉粥样硬化症、心脑血管疾病、肝癌或皮肤损伤。
  10. 一种如权利要求1-7任一项所述的化合物或其药学上可接受的盐或如权利要求8所述的药物组合物在制备用于抑制SREBP通路的药物中的应用。
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