WO2019020067A1 - 甾体类衍生物fxr激动剂的制备方法 - Google Patents

甾体类衍生物fxr激动剂的制备方法 Download PDF

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WO2019020067A1
WO2019020067A1 PCT/CN2018/097158 CN2018097158W WO2019020067A1 WO 2019020067 A1 WO2019020067 A1 WO 2019020067A1 CN 2018097158 W CN2018097158 W CN 2018097158W WO 2019020067 A1 WO2019020067 A1 WO 2019020067A1
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compound
formula
group
protecting group
tert
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PCT/CN2018/097158
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English (en)
French (fr)
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李小林
肖华玲
李鹏
贺海鹰
郝飞
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正大天晴药业集团股份有限公司
南京明德新药研发股份有限公司
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Priority to JP2020504103A priority Critical patent/JP7308811B2/ja
Priority to US16/633,122 priority patent/US11059854B2/en
Priority to EP18837478.9A priority patent/EP3660030A4/en
Priority to CN201880047098.2A priority patent/CN110869381B/zh
Publication of WO2019020067A1 publication Critical patent/WO2019020067A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J43/00—Normal steroids having a nitrogen-containing hetero ring spiro-condensed or not condensed with the cyclopenta(a)hydrophenanthrene skeleton
    • C07J43/003—Normal steroids having a nitrogen-containing hetero ring spiro-condensed or not condensed with the cyclopenta(a)hydrophenanthrene skeleton not condensed
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J51/00—Normal steroids with unmodified cyclopenta(a)hydrophenanthrene skeleton not provided for in groups C07J1/00 - C07J43/00
    • 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
    • C07J9/005—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 containing a carboxylic function directly attached or attached by a chain containing only carbon atoms to the cyclopenta[a]hydrophenanthrene skeleton
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the present application belongs to the field of medicinal chemistry, and in particular, the present application relates to a method for preparing a steroid derivative FXR agonist and related intermediates.
  • Farnesyl X receptor is an orphan nuclear receptor originally identified from rat liver cDNA libraries (BM. Forman, et al., Cell 81:687-693 (1995)), which is associated with insect molting. Hormone receptors are closely related. FXR is a member of the family of ligand-activated transcription factor nuclear receptors including steroids, retinoids, and thyroid hormone receptors (DJ. Mangelsdorf, et al., Cell 83: 841-850 (1995)). Northern and in situ analysis revealed extensive expression of FXR in the liver, intestine, kidney, and adrenal gland (BM. Forman et al., Cell 81:687-693 (1995) and W.
  • FXR Farnesyl X receptor
  • FXR forms a heterodimer with the 9-cis retinoic acid receptor (RXR) to bind to DNA.
  • RXR 9-cis retinoic acid receptor
  • the FXR/RXR heterodimer preferentially binds to a component consisting of a binuclear receptor half site of the consensus AG(G/T)TCA, which forms an inverted repeat and is separated by a single nucleoside (IR-1 motif) ( BM. Forman, et al., Cell 81: 687-693 (1995)).
  • IR-1 motif single nucleoside
  • the cholic acid as the FXR ligand includes chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), lithocholic acid (LCA), and taurine and glycine conjugates of these cholic acids.
  • the application provides a method of preparing a compound of formula I,
  • R 1 is selected from the group consisting of H, Cl, Br or F,
  • P 1 and P 2 are each independently selected from a hydroxy protecting group
  • M is selected from metal cations.
  • the present application provides a method of preparing a compound of formula 8, the method comprising the steps of:
  • Step g-1 the 7-position and 23-position hydroxyl groups of the compound of formula 7 are bonded to the protecting group P 2 to obtain the compound of the formula 12, and the step g-2) is carried out by removing the 23-hydroxy protecting group P 2 from the compound of the formula 12 to obtain a compound of formula 8,
  • R 2 is selected from the group consisting of C 1-6 alkyl groups
  • P 1 and P 2 are each independently selected from a hydroxy protecting group.
  • the P 1 and P 2 are each independently selected from a hydroxy protecting group that remains attached to the oxygen atom at position 3 or/and at position 7 of the above steroid compound under basic conditions, preferably at pH ⁇ 7.5, pH ⁇ 8, pH ⁇ 8.5, pH ⁇ 9, pH ⁇ 9.5, pH ⁇ 10, pH ⁇ 10.5, pH ⁇ 11, pH ⁇ 11.5 or pH ⁇ 12 under the conditions of the above steroid compound 3 or / and The 7-position oxygen atom remains attached to the hydroxy protecting group.
  • the P 1 and P 2 are each independently selected from a hydroxyl group that remains attached to the oxygen atom attached to the 3-position or/and the 7-position of the above-described steroid compound in the presence of a base or/and a protic solvent. Protection base.
  • the base is selected from a base having a pKa ⁇ 5, preferably pKa ⁇ 6, 50 ⁇ pKa ⁇ 8, 50 ⁇ pKa ⁇ 10, or 40 ⁇ pKa ⁇ 10; or the base is preferably selected from triethylamine ( TEA), diisopropylethylamine (DIPEA), 1,8-diazabicycloundec-7-ene (DBU), pyridine, 2,6-lutidine, 4-dimethylamino Pyridine (DMAP), imidazole, NaOH, KOH, LiOH, Mg(OH) 2 , Ca(OH) 2 , NaH, KH, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, sodium t-butoxide, potassium t-butoxide , KHMDS, NaHMDS, Na, K, KHCO 3 , NaHCO 3 , K 2 CO 3 or Na 2 CO 3
  • the protic solvent is selected from a C 1-6 alcohol or water, preferably from isopropanol, ethanol, methanol, n-propanol, n-butanol, tert-butanol or water.
  • the P 1 is selected from a silane ether protecting group, an alkyl ether protecting group, an alkoxy methyl ether protecting group or an ester protecting group; preferably a trimethylsilyl (TMS), Triethylsilyl (TES), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), benzyl, p-methoxybenzyl, 3,4-dimethoxy Benzyl, methyl ether, tert-butyl, trityl, 4,4'-dimethoxytrityl (DMTr), methoxymethyl (MOM), benzyloxymethyl (BnOM) , tetrahydrofuranyl (THP), formyl, acetyl (Ac), chloroacetyl (CAc), methoxyacetyl (MAc) or pivaloyl (Pv); more preferably trimethylsilyl (TMS
  • the P 2 is selected from a silane ether protecting group; preferably trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldimethylsilyl (TBDMS) or uncle Butyl diphenyl silicon (TBDPS); more preferably trimethylsilyl (TMS).
  • TMS trimethylsilyl
  • TES triethylsilyl
  • TDMS tert-butyldimethylsilyl
  • TDPS uncle Butyl diphenyl silicon
  • TMS trimethylsilyl
  • the M is selected from an alkali metal cation or an alkaline earth metal cation; preferably Li + , Na + , K + , Ca 2+ or Mg 2+ ; more preferably Na + or K + .
  • the R 2 is selected from C 1-4 alkyl, preferably methyl, ethyl, propyl or butyl.
  • the reaction is carried out in the presence of a base.
  • the base is selected from a base having a pKa ⁇ 12, preferably a base having a pKa ⁇ 15, a base having 50 ⁇ pKa ⁇ 15, a base having 40 ⁇ pKa ⁇ 15 or a base having 35 ⁇ pKa ⁇ 15;
  • the base is preferably selected from the group consisting of NaOH, KOH, LiOH, NaH, KH, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, sodium t-butoxide, potassium t-butoxide, KHMDS, NaHMDS, Na or K, more preferably sodium t-butoxide. Potassium tert-butoxide, KHMDS, NaHMDS, NaH, KH or Na, further preferably from sodium t-butoxide or potassium t-butoxide.
  • the solvent used in the reaction is selected from the group consisting of aprotic solvents; preferably ethers, ketones, nitriles, amides, sulfones, pyridine solvents; more preferably diethyl ether, isopropyl ether, Methyl tert-butyl ether, dibutyl ether, ethylene glycol diethyl ether, diethylene glycol dimethyl ether, tetrahydrofuran, 1,4-dioxane, acetone, methyl ethyl ketone, methyl isopropanone, cyclohexanone, methyl ketone , acetonitrile, propionitrile, dimethylformamide, dimethylacetamide, hexamethylphosphoramide, dimethyl sulfoxide or pyridine; further preferably tetrahydrofuran, 1,4-dioxane, dimethylformamide or Acetonitrile
  • the reaction temperature in step a), can be selected from a wide range. In some embodiments, the reaction temperature is selected from the group consisting of -30 ° C to 100 ° C, preferably -20 ° C to 80 ° C, more preferably -15 ° C to 70 ° C, further preferably -10 ° C to 60 ° C.
  • the reaction is carried out in the presence of a base.
  • the base is selected from a base having a pKa ⁇ 12, preferably a base having a pKa ⁇ 15, a base having 50 ⁇ pKa ⁇ 15, a base having 40 ⁇ pKa ⁇ 15 or a base having 35 ⁇ pKa ⁇ 15;
  • the base is preferably NaOH, KOH, LiOH, Mg(OH) 2 , Ca(OH) 2 , NaH, KH, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, sodium t-butoxide, potassium t-butoxide, KHMDS or NaHMDS is more preferably NaOH, KOH, LiOH or sodium methoxide.
  • the ratio of the compound of formula 10 to the amount of base charged is selected from 1:5 to 60, preferably 1:10 to 50, more preferably 1 : 15-40, further preferably 1:15-30; or, the ratio of the compound of the formula 10 to the amount of the base to be charged (in terms of molar amount) is preferably 1:5, 1:10, 1:15, 1: 20, 1:25, 1:35, 1:40, 1:45, 1:50, 1:55 or 1:60.
  • step b) the reaction is carried out in the presence of a protic solvent selected from the group consisting of C 1-6 alcohols or water, preferably from isopropanol, ethanol, methanol, n-propyl Alcohol, n-butanol, tert-butanol or water; preferably from ethanol or water.
  • a protic solvent selected from the group consisting of C 1-6 alcohols or water, preferably from isopropanol, ethanol, methanol, n-propyl Alcohol, n-butanol, tert-butanol or water; preferably from ethanol or water.
  • the reaction temperature is selected from the group consisting of 20 to 180 ° C; preferably 40 to 160 ° C; more preferably 60 to 140 ° C.
  • the reaction in step c), is carried out in the presence of an acid.
  • the acid is such that the reaction is carried out under conditions of pH ⁇ 6; preferably at pH ⁇ 5.5, pH ⁇ 5, pH ⁇ 4.5 or pH ⁇ 4.
  • the acid is selected from the group consisting of protic acids; preferably hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, formic acid, acetic acid, trifluoroacetic acid or p-toluenesulfonic acid; more preferably hydrochloric acid, Formic acid, acetic acid, trifluoroacetic acid or p-toluenesulfonic acid.
  • the solvent used in the reaction may be selected from a wide range, for example, may be selected from protic or/and aprotic solvents, preferably tetrahydrofuran, dichloromethane, acetonitrile, 1,4- Dioxane or dimethylformamide, methanol, ethanol, n-propanol, isopropanol, n-butanol, tert-butanol or water.
  • protic or/and aprotic solvents preferably tetrahydrofuran, dichloromethane, acetonitrile, 1,4- Dioxane or dimethylformamide, methanol, ethanol, n-propanol, isopropanol, n-butanol, tert-butanol or water.
  • the reaction temperature is selected from the group consisting of -30 ° C to 100 ° C; preferably -15 ° C to 80 ° C.
  • the alcohol is selected from the group consisting of C1-6 alcohols; preferably C1-4 alcohols; more preferably methanol, ethanol, propanol or butanol.
  • the reaction in step d), is carried out in the presence of an acid.
  • the acid is selected from a protic acid or a Lewis acid; preferably hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, formic acid, acetic acid, trifluoroacetic acid, p-toluenesulfonic acid, AlCl 3 , FeCl 3 or BF 3 ; more preferably from hydrochloric acid, sulfuric acid or p-toluenesulfonic acid.
  • the reaction temperature is selected from the group consisting of 40 ° C to 120 ° C, preferably 60 ° C to 100 ° C.
  • the reaction in step e), is carried out in the presence of a base.
  • the base is selected from the group consisting of triethylamine (TEA), diisopropylethylamine (DIPEA), 1,8-diazabicycloundec-7-ene (DBU), pyridine , 2,6-lutidine, 4-dimethylaminopyridine (DMAP), imidazole, Na 2 CO 3 , K 2 CO 3 , Cs 2 CO 3 , NaH or KH; preferably TEA, DIPEA, DBU, pyridine, Imidazole, Na 2 CO 3 or K 2 CO 3 .
  • the ratio of the compound of formula 5 to the amount of base charged is selected from 1:0.5-10, preferably from 1:0.8-8, more preferably From 1:1-6.
  • the solvent used in the reaction is selected from the group consisting of aprotic solvents, preferably diethyl ether, diisopropyl ether, methyl tert-butyl ether, dibutyl ether, ethylene glycol diethyl ether, diethylene glycol Methyl ether, THF, 1,4-dioxane, DCM, ethyl acetate, CH 3 CN, toluene, xylene, acetone, methyl ethyl ketone, methyl isopropanone, cyclohexanone, methyl ketone, DMF or DMSO, Preference is given to THF, DCM, ethyl acetate, CH 3 CN, toluene, acetone or DMF.
  • aprotic solvents preferably diethyl ether, diisopropyl ether, methyl tert-butyl ether, dibutyl ether, ethylene glycol dieth
  • the reaction temperature is selected from -10 °C to 100 °C, preferably from 0 °C to 60 °C.
  • the reaction in step f), is carried out in the presence of a reducing agent.
  • the reducing agent is selected from the group consisting of lithium aluminum hydride, diisobutylaluminum hydride (DIBAL-H), red aluminum (Red-Al), BH 3 , NaBH 4 , KBH 4 or Zn (BH 4 2 , lithium tetrahydrogenate is preferred.
  • the solvent used in the reaction is selected from a protic or aprotic solvent, preferably THF, dioxane, methanol, ethanol or tert-butyl methyl ether.
  • the reaction temperature is selected from the group consisting of -30 ° C to 80 ° C, preferably -15 ° C to 60 ° C.
  • the reaction is carried out in the presence of a base.
  • the base is selected from a base having a pKa ⁇ 5, preferably pKa ⁇ 6, pKa ⁇ 8, pKa ⁇ 10, 30 ⁇ pKa ⁇ 5, 20 ⁇ pKa ⁇ 5, 30 ⁇ pKa ⁇ 6, 30 ⁇ a base having pKa ⁇ 8, 30 ⁇ pKa ⁇ 10, 20 ⁇ pKa ⁇ 6, 20 ⁇ pKa ⁇ 8, 20 ⁇ pKa ⁇ 10; in some embodiments, the base is selected from the group consisting of triethylamine (TEA), diiso) Propylethylamine (DIPEA), 1,8-diazabicycloundec-7-ene (DBU), pyridine, 2,6-lutidine, 4-dimethylaminopyridine (DMAP) or Imidazole, preferably triethylamine
  • the reaction in step g-2), is carried out at a pH ⁇ 7. In some embodiments, preferably, the reaction is carried out under conditions of pH ⁇ 7.5, pH ⁇ 8 or pH ⁇ 8.5.
  • the base used to adjust the pH may be selected from KHCO 3 , NaHCO 3 , K 2 CO 3 or Na 2 CO 3 , preferably K 2 CO 3 or Na 2 CO 3 .
  • the pH-adjusted base in step g-2), can be selected from a wide range, and the reaction is allowed to proceed under suitable pH conditions.
  • the ratio of the compound of formula 7 to the pH-adjusting base is selected from the group consisting of 1:0.5-10, preferably 1:1-8, more preferably 1:2-6.
  • the solvent used in the reaction is selected from the group consisting of an aprotic solvent, preferably diethyl ether, diisopropyl ether, methyl tert-butyl ether, dibutyl ether, ethylene glycol diethyl ether, diethylene glycol Alcohol, THF, 1,4-dioxane, DCM, ethyl acetate, CH 3 CN, toluene, xylene, acetone, methyl ethyl ketone, methyl isopropanone, cyclohexanone, methyl ketone, DMF, DMSO N-hexane or cyclohexane, more preferably DCM, THF, toluene, n-hexane or cyclohexane.
  • an aprotic solvent preferably diethyl ether, diisopropyl ether, methyl tert-butyl ether, dibutyl ether, ethylene glycol
  • step g-2 the reaction is carried out in the presence of a protic solvent selected from the group consisting of C 1-6 alcohols or water, preferably from isopropanol, ethanol, methanol, N-propanol, n-butanol, tert-butanol or water, more preferably methanol or ethanol.
  • a protic solvent selected from the group consisting of C 1-6 alcohols or water, preferably from isopropanol, ethanol, methanol, N-propanol, n-butanol, tert-butanol or water, more preferably methanol or ethanol.
  • the application provides a compound of formula 6, a compound of formula 7, a compound of formula 8, a compound of formula 10, and a compound of formula 11,
  • R 1 is selected from the group consisting of H, Cl, Br or F,
  • R 2 is selected from the group consisting of methyl, ethyl, propyl or butyl.
  • P 1 is selected from a silyl ether protecting group, an alkyl ether protecting group, an alkoxy methyl ether protecting group or an ester protecting group.
  • P 2 is selected from a silane ether protecting group
  • M is selected from metal cations.
  • the application provides the use of a compound of formula 6 and a compound of formula 7 in the preparation of a compound of formula 8 or a compound of formula I,
  • R 1 is selected from the group consisting of H, Cl, Br or F,
  • the R 2 is selected from the group consisting of methyl, ethyl, propyl or butyl.
  • the P 1 is selected from a silyl ether protecting group, an alkyl ether protecting group, an alkoxy methyl ether protecting group or an ester protecting group.
  • the P 2 is selected from a silane ether-based protecting group.
  • the application provides the use of a compound of formula 8, a compound of formula 10, and a compound of formula 11 in the preparation of a compound of formula I,
  • R 1 is selected from the group consisting of H, Cl, Br or F,
  • the P 1 is selected from a silyl ether protecting group, an alkyl ether protecting group, an alkoxy methyl ether protecting group or an ester protecting group.
  • the P 2 is selected from a silane ether protecting group
  • the M is selected from a metal cation.
  • the M is selected from an alkali metal cation or an alkaline earth metal cation; preferably Li + , Na + , K + , Ca 2+ or Mg 2+ ; more preferably Na + or K + .
  • the P 1 is selected from the group consisting of a silyl ether protecting group, an alkyl ether protecting group, an alkoxymethyl ether protecting group or an ester protecting group; Methylsilyl (TMS), triethylsilyl (TES), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), benzyl, p-methoxybenzyl , 3,4-dimethoxybenzyl, methyl ether, tert-butyl, trityl, 4,4'-dimethoxytrityl (DMTr), methoxymethyl (MOM) , benzyloxymethyl (BnOM), tetrahydrofuranyl (THP), formyl, acetyl (Ac), chloroacetyl (CAc), methoxyacetyl (MAc) or pivaloyl
  • TMS Methylsilyl
  • the P 2 is selected from a silane ether protecting group; preferably a trimethylsilyl (TMS), a triethylsilyl (TES), a tert-butyl dimethyl group Silicon-based (TBDMS) or tert-butyldiphenylsilyl (TBDPS); more preferably trimethylsilyl (TMS).
  • TMS trimethylsilyl
  • TES triethylsilyl
  • TDMS tert-butyl dimethyl group Silicon-based
  • TDPS tert-butyldiphenylsilyl
  • the compound of Formula 6 is selected from the group consisting of
  • the compound of Formula 7 is selected from the group consisting of
  • the compound of Formula 8 is selected from the group consisting of
  • the compound of Formula 10 is selected from the group consisting of
  • the compound of Formula 11 is selected from the group consisting of
  • the alkyl group includes all of its isomeric forms, for example, propyl includes n-propyl, isopropyl; butyl includes 1-n-butyl, 2-n-butyl, iso Butyl, tert-butyl.
  • the protecting group and its attachment or removal method can be carried out by a conventional method in the art, and the method can be a one-step reaction or a multi-step reaction, such as, but not limited to, referring to "Greene's of Wiley Press”. Implementation of Protective Groups in Organic Synthesis-4th Edition or Chemical Industry Press.
  • the compounds may exist in specific geometric or stereoisomeric forms.
  • This application contemplates all such compounds, including the cis and trans isomers, the (-)- and (+)-p-enantiomers, the (R)- and (S)-enantiomers, diastereomeric a conformation, a (D)-isomer, a (L)-isomer, and a racemic mixture thereof, and other mixtures, such as enantiomeric or diastereomeric excess mixtures, all of which belong to the present Within the scope of the application. Additional asymmetric carbon atoms may be present in the substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the invention.
  • the atoms of the steroidal compounds can be numbered according to the general knowledge in the art, such as, but not limited to, the reference to Xing Qiyi, edited by the Higher Education Press, Basic Organic Chemistry - Third Edition.
  • the compound of formula 7 is taken as an example, and the atoms of 3, 7, and 23 are labeled:
  • reaction conditions of the production method of the present application are mild, and can be carried out, for example, under normal pressure, and the reaction temperature is easily controlled. Moreover, some steps in the preparation method described in the present application can simultaneously convert a plurality of groups, effectively shortening the steps, and is suitable for industrial production.
  • the solution was evaporated to dryness to give a crude material, which was then dissolved in ethyl acetate (30 L), followed by saturated sodium hydrogen carbonate solution (9 L), water (9 L), saturated saline (9 L) was washed. The organic phase is spin dried to give the product.
  • the product trait is a brown oily liquid.
  • the compound of the formula 5a (3100 g) was dissolved in dichloromethane (30 L), and then imidazole (529.4 g) and triethylamine (786.8 g) were added, and the temperature was lowered in the kettle (internal temperature was 5 ° C), and slowly at this temperature.
  • TBDPSCl (2140 g) was added dropwise, and the temperature during the dropwise addition did not exceed 10 ° C. After the dropwise addition was completed, the reaction was stirred at room temperature for 16 hours. TLC showed that the starting material was completely reacted, and 15 L of water was slowly added dropwise to the reaction mixture to quench the reaction. The mixture was allowed to stand, and the reaction was separated. The methylene chloride phase was separated, and the methylene chloride phase was washed with saturated brine (10 L).
  • the product trait is a brown oily liquid.
  • Tetrahydrofuran (10 L) was added to a 50 L autoclave at 15 ° C, and LiAlH 4 (235 g, 6.2 mol) was added to the reaction vessel under N 2 protection, and the reaction was cooled to an internal temperature of 5 °C. After dissolving the compound of the formula 6a (2.04 kg) in tetrahydrofuran, it was slowly added dropwise to a solution of LiAlH 4 in tetrahydrofuran for about 2.5 hours. The reaction was stirred at 15 ° C for 2 hours, and the reaction was monitored by TLC to show that material disappeared.
  • reaction mixture was slowly added dropwise to the reaction solution with H 2 O (235 mL), and then a tetrahydrofuran solution (20 L) was added to the reaction solution, and then 15% NaOH solution (235 mL) was slowly added dropwise to the reaction solution, and stirred. 12 hours.
  • the reaction mixture was filtered, and the filter cake was washed with dichloromethane (3L). After dissolving the oil in DCM (15 L), the organic phase was washed once with water (5L), brine (5L), and the filtrate was evaporated to give a white solid (1.8 kg).
  • the reaction mixture was cooled to room temperature (about 16 ° C) Then, the pH is adjusted to about 9 with a saturated sodium hydrogen carbonate solution, and the solution is dried (small amount) to give a crude product.
  • the crude product is dissolved in ethyl acetate (30 L), followed by saturated sodium bicarbonate (9 L), water ( 9L), saturated saline solution (9L) was washed.
  • the organic phase is spin dried to give the product.
  • the product trait is a brown oily liquid.
  • the organic phase was concentrated to about 5 liters and 30 liters of ethanol was added to the solution. Potassium carbonate (1.93 kg, 13.95 mol) was added to the solution at 15 °C. The system was stirred at 15 ° C for 14 hours. The TLC detection reaction was almost complete. The reaction solution was filtered. The filter cake was rinsed with 3 liters of dichloromethane. The filtrate was concentrated to give an oil. The oil was dissolved in 20 liters of dichloromethane and washed once with 10 liters of water and 10 liters of brine. The organic phase was dried over 3 kg of anhydrous sodium sulfate and filtered.
  • the compound of formula 8a (2498.0 g, 3.10 mol) was added to the reaction vessel, dissolved in THF (12.5 L), the internal temperature was controlled from 5 ° C to 10 ° C, t-BuONa (614.2 g, 6.20 mol) was slowly added, and the addition was completed in about 40 min. After stirring for 10 min, the temperature was raised to 20 ° C to 23 ° C and stirred for 1.5 h, and the temperature was lowered to 5 ° C to 10 ° C. While maintaining the internal temperature, a THF solution (12.5 L, 6.20 mol, 1073.1 g) of the compound of the formula 9a was added dropwise to the reaction mixture, and the mixture was heated to 60 °C.
  • the compound of formula 10a (2520.0 g, 2.79 mol) was added to the reaction vessel (20 L), EtOH (13.0 L) was added, stirred and dissolved, the internal temperature was controlled to 10 ° C, and an aqueous solution of NaOH (2232.0 g, 55.8 mol) (13.0 L) was added. , joined in batches. The temperature was raised to 105 ° C and stirred for 2.8 hours. The reaction was complete by TLC and HPLC. The reaction solution was cooled to 10 ° C and allowed to stand for two hours, and a solid was precipitated from the bottom of the bottle.
  • the compound of the formula 11a (2330.3 g, 2.65 mmol) was added to a reaction mixture (50 L), and THF (24.0 L) was dissolved. The internal temperature was controlled at 10 ° C, and a concentrated HCl (10.0 L, 120.00 mol) liquid was slowly added dropwise thereto, and the mixture was heated to 13 ° C (room temperature) for 2 hours under stirring for 90 hours.
  • Protein glutathione-S-transferase-labeled FXR human protein (Invitrogen)
  • Coactivator biotinylated steroid receptor coactivator (Anaspec)
  • Detection reagent homogeneous proximity luminescence amplification test (alphascreen) detection kit (PerkinElmer)
  • test compound was prepared as a 40 ⁇ M DMSO solution, and then the compound was diluted 3-fold to 10 concentration points.
  • the reference compound was prepared as a 400 ⁇ M DMSO solution, followed by dilution at 1.5 fold to 10 concentration points.
  • the diluted DMSO solution was added to the wells of a 384-well plate at a volume of 150 nL per well.
  • the glutathione-S-transferase-labeled FXR human protein and the biotin-labeled steroid receptor coactivator were formulated into a mixed solution having a concentration of 0.4 nM and 30 nM, respectively.
  • the wells of the 384-well plate were added in a volume of 15 ⁇ L per well. Incubate for 1 hour at room temperature.
  • the receptor pellet mixture in the homogeneous proximity luminescence amplification assay (alphascreen) assay kit was diluted 125-fold and added to the wells of a 384-well plate at a volume of 7.5 ⁇ L per well. The experiment process is protected from light. Incubate for 1 hour at room temperature.
  • the donor pellet mixture in the homogeneous proximity luminescence amplification assay (alphascreen) assay kit was diluted 125-fold and added to the wells of a 384-well plate at a volume of 7.5 ⁇ L per well. The experiment process is protected from light. Incubate for 1 hour at room temperature.
  • Cell culture medium 10% serum and Penicillin/Streptomycin (1 ⁇ ) were added to DMEM medium.
  • test compound was prepared as a 100 ⁇ M DMSO solution, and then the compound was diluted 3-fold to 10 concentration points.
  • the reference compound was prepared as a 100 ⁇ M DMSO solution, followed by dilution at 1.3 fold to 10 concentration points.
  • the diluted DMSO solution was added to the wells of a 384-well plate at a volume of 200 nL per well.
  • FXR HEK 293T DA cells were resuspended, resuspended in medium, diluted to a density of 5 ⁇ 10 5 /mL, and added to the wells of a 384-well plate at a volume of 40 ⁇ L per well.
  • the 384 microplate was incubated for 16 hours at 37 ° C under 5% CO 2 .
  • the mixed solution B C 1mM LiveBLAzer 6 ⁇ L of TM -FRET B / G (CCF4- AM) and the substrate with 60 ⁇ L 934 ⁇ L of volume per well was added 8 ⁇ L micropores 384 well plate.
  • Test sample FXR enzyme activity
  • FXR cell activity FXR cell activity
  • the compounds of the present application have significant agonistic effects on the FXR receptor, and the agonistic effect on the FXR receptor is also significant at the cellular level.
  • mice Twelve C57BL/6J male mice were randomly divided into two groups, 6 in each group.
  • the first group was intravenous group.
  • the solvent is 10% HPbCD aqueous solution, when the drug solubility is not ideal, the help solvent will be added
  • the second group is oral group, irrigation
  • the stomach was administered at 10 mg/kg and 10 mL/kg (the vehicle was a 0.5% HPMC aqueous solution).
  • Plasma (K 2 -EDTA is anticoagulated) samples were taken at 0.083, 0.25, 0.5, 1, 2, 4, 6 , 8 and 24 hours after intravenous administration; 0.25, 0.5, 1, 2, 4 after oral administration Plasma samples were taken at 6, 8 and 24 hours.
  • the peak concentration of the compound of formula I-H after oral administration of the same dose was similar to that of the control compound obeticholic acid, and the drug exposure was higher than that of the control compound obeticholic acid.
  • the peak concentration of the compound of formula I-Cl after oral administration of the same dose was higher than that of the control compound obeticholic acid, and the drug exposure was also higher than that of the control compound obeticholic acid.
  • mice Six C57BL/6J male mice were grouped into the oral group.
  • the preparation contained 5 kinds of research and development drugs, and 2 mg/kg/compound was administered by intragastric administration (the solvent was 0.5% HPMC aqueous solution).
  • the five compounds were first dissolved in a solvent, respectively, and formed into a 1 mg/mL solution (clear solution or suspension) by ultrasonic or vortexing, and then the five compound solutions were mixed in equal volumes (1:1:1:1:1). v:v:v:v:v:v) in a glass bottle.
  • 3 animals were collected for plasma and liver tissue samples 0.5 hours after administration; the other 3 animals were collected corresponding samples 3 hours after administration.

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Abstract

属于药物化学领域，涉及甾体类衍生物FXR激动剂的制备方法及相关中间体。具体而言，涉及一种制备式I化合物的方法，其包括使式8化合物和式9化合物反应以获得式10化合物，从式10化合物反应以获得式11化合物，从式11化合物反应以获得式I化合物，以及所用中间体、中间体的制备方法和中间体的用途。该制备方法反应条件温和，部分步骤可以同时转变多个基团，有效缩短步骤，适合工业化生产。 (I)

Description

甾体类衍生物FXR激动剂的制备方法 技术领域
本申请属于药物化学领域，具体而言，本申请涉及甾体类衍生物FXR激动剂的制备方法及相关中间体。
背景技术
法尼酯X受体(FXR)是一种最初从大鼠肝cDNA文库中鉴定的孤儿核受体(BM.Forman，et al.，Cell 81：687-693(1995))，其与昆虫蜕皮激素受体密切相关。FXR是包括类固醇、类维生素A、和甲状腺激素受体的配基-激活转录因子核受体家族的成员(DJ.Mangelsdorf，et al.，Cell 83：841-850(1995))。Northern和原位分析显示FXR在肝、肠、肾和肾上腺中大量表达(BM.Forman et al.，Cell 81：687-693(1995)和W.Seol et al.，Mol.Endocrinnol,9：72-85(1995))。FXR与9-顺式维生素A酸受体(RXR)形成异源二聚体与DNA结合。FXR/RXR异源二聚体优先与由共有AG(G/T)TCA的双核受体半位点组成的成分结合，其形成反向重复并被单一核苷分离(IR-1模体)(BM.Forman，et al.，Cell 81：687-693(1995))。然而，这些化合物无法激活小鼠和人类FXR，使得内源性FXR配基的自然性还不确定。一些自然发生的胆酸在生理浓度下结合并激活FXR(PCTWO 2000/37077，2000年6月29日出版))。如此所述，作为FXR配基的胆酸包括鹅脱氧胆酸(CDCA)、脱氧胆酸(DCA)、石胆酸(LCA)和这些胆酸的牛磺酸及氨基乙酸共轭物。
发明内容
一方面，本申请提供一种制备式I化合物的方法，
Figure PCTCN2018097158-appb-000001
包括如下步骤：
a)使式8化合物和式9化合物反应以获得式10化合物，
Figure PCTCN2018097158-appb-000002
b)从式10化合物反应以获得式11化合物，
Figure PCTCN2018097158-appb-000003
c)从式11化合物反应以获得式I化合物，
Figure PCTCN2018097158-appb-000004
其中，R 1选自H、Cl、Br或F，
P 1和P 2各自独立地选自羟基保护基，
M选自金属阳离子。
另一方面，本申请提供一种制备式8化合物的制备方法，所述方法包括如下步骤：
d)以式4化合物为原料，与醇发生酯化反应以获得式5化合物，
Figure PCTCN2018097158-appb-000005
e)式5化合物3位羟基连接保护基以获得式6化合物，
Figure PCTCN2018097158-appb-000006
f)从式6化合物反应以获得式7化合物，
Figure PCTCN2018097158-appb-000007
g)步骤g-1)，式7化合物7位和23位羟基连接保护基P 2以获得式12化合物，步骤g-2)，从式12化合物反应脱除23位羟基保护基P 2以获得式8化合物，
Figure PCTCN2018097158-appb-000008
其中，R 2选自C 1-6烷基，
P 1和P 2各自独立地选自羟基保护基。
在一些实施方案中，所述P 1和P 2各自独立地选自在碱性条件下与上述甾体化合物3位或/和7位所连氧原子保持连接的羟基保护基，优选在pH≥7.5、pH≥8、pH≥8.5、pH≥9、pH≥9.5、pH≥10、pH≥10.5、pH≥11、pH≥11.5或pH≥12的条件下与上述甾体化合物3位或/和7位所连氧原子保持连接的羟基保护基。
在一些实施方案中，所述P 1和P 2各自独立地选自在碱或/和质子性溶剂存在的条件下与上述甾体化合物3位或/和7位所连氧原子保持连接的羟基保护基。在一些实施方案中，所述碱选自pKa≥5，优选pKa≥6、50≥pKa≥8、50≥pKa≥10或40≥pKa≥10的碱；或者所述碱优选自三乙胺(TEA)、二异丙基乙基胺(DIPEA)、1,8-二氮杂二环十一碳-7-烯(DBU)、吡啶、2,6-二甲基吡啶、4-二甲氨基吡啶(DMAP)、咪唑、NaOH、KOH、LiOH、Mg(OH) 2、Ca(OH) 2、NaH、KH、甲醇钠、甲醇钾、乙醇钠、乙醇钾、叔丁醇钠、叔丁醇钾、KHMDS、NaHMDS、Na、K、KHCO 3、NaHCO 3、K 2CO 3或Na 2CO 3。在一些实施方案中，所述质子性溶剂选自C 1-6醇或水，优选自异丙醇、乙醇、甲醇、正丙醇、正丁醇、叔丁醇或水。
在一些实施方案中，所述P 1选自硅烷醚类保护基、烷基醚类保护基、烷氧基甲基醚类保护基或酯类保护基；优选三甲基硅基(TMS)、三乙基硅基(TES)、叔丁基二甲基硅基(TBDMS)、叔丁基二苯基硅基(TBDPS)、苄基、对甲氧基苄基、3,4-二甲氧基苄基、甲基醚、叔丁基、三苯甲基、4,4’-二甲氧基三苯甲基(DMTr)、甲氧基甲基(MOM)、苄氧基甲基(BnOM)、四氢呋喃基(THP)、甲酰基、乙酰基(Ac)、氯乙酰基(CAc)、甲氧基乙酰基(MAc)或新戊酰基(Pv)；更优选三甲基硅基(TMS)、三乙基硅基(TES)、叔丁基二甲基硅基(TBDMS)、叔丁基二苯基硅基(TBDPS)或甲氧基甲基(MOM)。
在一些实施方案中，所述P 2选自硅烷醚类保护基；优选三甲基硅基(TMS)、三乙基硅基(TES)、叔丁基二甲基硅基(TBDMS)或叔丁基二苯基硅基(TBDPS)；更优选三甲基硅基(TMS)。
在一些实施方案中，所述M选自碱金属阳离子或碱土金属阳离子；优选Li +、Na +、K +、Ca 2+或Mg 2+；更优选Na +或K +。
在一些实施方案中，所述R 2选自C 1-4烷基，优选甲基、乙基、丙基或丁基。
在一些实施方案中，在步骤a)中，反应在碱的存在下进行。在一些实施方案中，所述碱选自pKa≥12的碱，优选pKa≥15的碱、50≥pKa≥15的碱、40≥pKa≥15的碱或35≥pKa≥15的碱；或者所述碱优选自NaOH、KOH、LiOH、NaH、KH、甲醇钠、甲醇钾、乙醇钠、乙醇钾、叔丁醇钠、叔丁醇钾、KHMDS、NaHMDS、Na或K，更优选叔丁醇钠、叔丁醇钾、KHMDS、NaHMDS、NaH、KH或Na，进一步优选自叔丁醇钠或叔丁醇钾。
在一些实施方案中，在步骤a)中，反应所用溶剂选自非质子性溶剂；优选醚类、酮类、腈类、酰胺类、砜类、吡啶类溶剂；更优选乙醚、异丙醚、甲基叔丁基醚、丁醚、乙二醇二乙醚、二乙二醇二甲醚、四氢呋喃、1,4-二氧六环、丙酮、甲乙酮、甲异丙酮、环己酮、甲己酮、乙腈、丙腈、二甲基甲酰胺、二甲基乙酰胺、六甲基磷酰胺、二甲亚砜或吡啶；进一步优选四氢呋喃、1,4-二氧六环、二甲基甲酰胺或乙腈。
在一些实施方案中，在步骤a)中，反应温度可选自较宽的范围。在一些实施方案中，反应温度选自-30℃至100℃，优选-20℃至80℃，更优选-15℃至70℃，进一步优选-10℃至60℃。
在一些实施方案中，在步骤b)中，反应在碱的存在下进行。在一些实施方案中，所述碱选自pKa≥12的碱，优选pKa≥15的碱、50≥pKa≥15的碱、40≥pKa≥15的碱或35≥pKa≥15的碱；或者，所述碱优选NaOH、KOH、LiOH、Mg(OH) 2、Ca(OH) 2、NaH、KH、甲醇钠、甲醇钾、乙醇钠、乙醇钾、叔丁醇钠、叔丁醇钾、KHMDS或NaHMDS，更优选NaOH、KOH、LiOH或甲醇钠。在一些实施方案中，在步骤b)中，所述式10化合物与所述碱的投料量(以摩尔量计)之比选自1:5-60，优选1:10-50，更优选1:15-40，进一步优选1:15-30；或者，所述式10化合物与所述碱的投料量(以摩尔量计)之比优选1:5、1:10、1:15、1:20、1:25、1:35、1:40、1:45、1:50、1:55或1:60。
在一些实施方案中，在步骤b)中，反应在质子性溶剂存在的条件下进行，所述质子性溶剂选自C 1-6醇或水，优选自异丙醇、乙醇、甲醇、正丙醇、正丁醇、叔丁醇或水；优选自乙醇或水。
在一些实施方案中，在步骤b)中，反应温度选自20至180℃；优选40至160℃；更优选60至140℃。
在一些实施方案中，在步骤c)中，反应在酸的存在下进行。在一些实施方案中，所述酸使得反应在pH≤6的条件下进行；优选在pH≤5.5、pH≤5、pH≤4.5或pH≤4的条件下进行。在一些实施方案中，在步骤c)中，所述酸选自质子酸；优选盐酸、氢溴酸、氢氟酸、硫酸、甲酸、乙酸、三氟乙酸或对甲苯磺酸；更优选盐酸、甲酸、乙酸、三氟乙酸或对甲苯磺酸。
在一些实施方案中，在步骤c)中，反应所用溶剂可选自较宽的范围，例如可选自质子性或/和非质子性溶剂，优选四氢呋喃、二氯甲烷、乙腈、1,4-二氧六环或二甲基甲酰胺、甲醇、乙醇、正丙醇、异丙醇、正丁醇、叔丁醇或水。
在一些实施方案中，在步骤c)中，反应温度选自-30℃至100℃；优选-15℃至80℃。
在一些实施方案中，在步骤d)中，所述醇选自C 1-6的醇；优选C 1-4的醇；更优选甲醇、乙醇、丙醇或丁醇。
在一些实施方案中，在步骤d)中，反应在酸的存在下进行。在一些实施方案中，所述酸选自质子酸或路易斯酸；优选盐酸、氢溴酸、氢氟酸、硫酸、甲酸、乙酸、三氟乙酸、对甲苯磺酸、AlCl 3、FeCl 3或BF 3；更优选自盐酸、硫酸或对甲苯磺酸。
在一些实施方案中，在步骤d)中，反应温度选自40℃至120℃，优选60℃至100℃。
在一些实施方案中，在步骤e)中，反应在碱的存在下进行。在一些实施方案中，所述碱选自三乙胺(TEA)、二异丙基乙基胺(DIPEA)、1,8-二氮杂二环十一碳-7-烯(DBU)、吡啶、2,6-二甲基吡啶、4-二甲氨基吡啶(DMAP)、咪唑、Na 2CO 3、K 2CO 3、Cs 2CO 3、NaH或KH；优选TEA、DIPEA、DBU、吡啶、咪唑、Na 2CO 3或K 2CO 3。在一些实施方案中，在步骤e)中，所述式5化合物与所述碱的投料量(以摩尔量计)之比选自1:0.5-10，优选自1:0.8-8，更优选自1:1-6。
在一些实施方案中，在步骤e)中，反应所用溶剂选自非质子性溶剂，优选乙醚、异丙醚、甲基叔丁基醚、丁醚、乙二醇二乙醚、二乙二醇二甲醚、THF、1,4-二氧六环、DCM、乙酸乙酯、CH 3CN、甲苯、二甲苯、丙酮、甲乙酮、甲异丙酮、环己酮、甲己酮、DMF或DMSO，更优选THF、DCM、乙酸乙酯、CH 3CN、甲苯、丙酮或DMF。
在一些实施方案中，在步骤e)中，反应温度选自-10℃至100℃，优选0℃至60℃。
在一些实施方案中，在步骤f)中，反应在还原剂的存在下进行。在一些实施方案中，所述还原剂选自四氢铝锂、二异丁基氢化铝(DIBAL-H)、红铝(Red-Al)、BH 3、NaBH 4、KBH 4或Zn(BH 4) 2，优选四氢铝锂。
在一些实施方案中，在步骤f)中，反应所用溶剂选自质子性或非质子性溶剂，优选THF、二氧六环、甲醇、乙醇或叔丁基甲醚。
在一些实施方案中，在步骤f)中，反应温度选自-30℃至80℃，优选-15℃至60℃。
在一些实施方案中，在步骤g-1)中，反应在碱的存在下进行。在一些实施方案中，所述碱选自pKa≥5的碱，优选pKa≥6、pKa≥8、pKa≥10、30≥pKa≥5、20≥pKa≥5、30≥pKa≥6、30≥pKa≥8、30≥pKa≥10、20≥pKa≥6、20≥pKa≥8、20≥pKa≥10的碱；在部分实施方式中，所述碱选自三乙胺(TEA)、二异丙基乙基胺(DIPEA)、1,8-二氮杂二环十一碳-7-烯(DBU)、吡啶、2,6-二甲基吡啶、4-二甲氨基吡啶(DMAP)或咪唑，优选三乙胺(TEA)、二异丙基乙基胺(DIPEA)、吡啶或咪唑。
在一些实施方案中，在步骤g-2)中，反应在pH≥7的条件下进行。在一些实施方案中，优选地，所述反应在pH≥7.5、pH≥8或pH≥8.5的条件下进行。在一些实施方案中，用于调节pH的碱可选自KHCO 3、NaHCO 3、K 2CO 3或Na 2CO 3，优选K 2CO 3或Na 2CO 3。在一些实施方案中，在步骤g-2)中，所述调节pH的碱的投料量可选自较宽的范围，满足反应在合适的pH条件下进行即可。在一些实施方案中，所述式7化合物与调节pH的碱的投料量(以摩尔量计)之比选自1:0.5-10，优选1:1-8，更优选1:2-6。
在一些实施方案中，在步骤g-1)中，反应所用溶剂选自非质子性溶剂，优选乙醚、异丙醚、甲基叔丁基醚、丁醚、乙二醇二乙醚、二乙二醇二甲醚、THF、1,4-二氧六环、DCM、乙酸乙酯、CH 3CN、甲苯、二甲苯、丙酮、甲乙酮、甲异丙酮、环己酮、甲己酮、DMF、DMSO、正己烷或环己烷，更优选DCM、THF、甲苯、正己烷或环己烷。
在一些实施方案中，在步骤g-2)中，反应在质子性溶剂存在的条件下进行，所述质子性溶剂选自C 1-6醇或水，优选自异丙醇、乙醇、甲醇、正丙醇、正丁醇、叔丁醇或水，更优选甲醇或乙醇。
中间体
另一方面，本申请提供如下式6化合物、式7化合物、式8化合物、式10化合物和式11化合物，
Figure PCTCN2018097158-appb-000009
其中，R 1选自H、Cl、Br或F，
R 2选自甲基、乙基、丙基或丁基，
P 1选自硅烷醚类保护基、烷基醚类保护基、烷氧基甲基醚类保护基或酯类保护基，
P 2选自硅烷醚类保护基，
M选自金属阳离子。
另一方面，本申请提供如下式6化合物和式7化合物在制备式8化合物或式I化合物中的用途，
Figure PCTCN2018097158-appb-000010
其中，所述R 1选自H、Cl、Br或F，
所述R 2选自甲基、乙基、丙基或丁基，
所述P 1选自硅烷醚类保护基、烷基醚类保护基、烷氧基甲基醚类保护基或酯类保护基，
所述P 2选自硅烷醚类保护基。
另一方面，本申请提供如下式8化合物、式10化合物和式11化合物在制备式I化合物中的用途，
Figure PCTCN2018097158-appb-000011
其中，所述R 1选自H、Cl、Br或F，
所述P 1选自硅烷醚类保护基、烷基醚类保护基、烷氧基甲基醚类保护基或酯类保护基，
所述P 2选自硅烷醚类保护基，
所述M选自金属阳离子。
在一些实施方案中，上述中间体或其用途中，所述M选自碱金属阳离子或碱土金属阳离子；优选Li +、Na +、K +、Ca 2+或Mg 2+；更优选Na +或K +。
在一些实施方案中，上述中间体或其用途中，所述P 1选自硅烷醚类保护基、烷基醚类保护基、烷氧基甲基醚类保护基或酯类保护基；优选三甲基硅基(TMS)、三乙基硅基(TES)、叔丁基二甲基硅基(TBDMS)、叔丁基二苯基硅基(TBDPS)、苄基、对甲氧基苄基、3,4-二甲氧基苄基、甲基醚、叔丁基、三苯甲基、4,4’-二甲氧基三苯甲基(DMTr)、甲氧基甲基(MOM)、苄氧基甲基(BnOM)、四氢呋喃基(THP)、甲酰基、乙酰基(Ac)、氯乙酰基(CAc)、甲氧基乙酰基(MAc)或新戊酰基(Pv)；更优选三甲基硅基(TMS)、三乙基硅基(TES)、叔丁基二甲基硅基(TBDMS)、叔丁基二苯基硅基(TBDPS)或甲氧基甲基(MOM)。
在一些实施方案中，上述中间体或其用途中，所述P 2选自硅烷醚类保护基；优选三甲基硅基(TMS)、三乙基硅基(TES)、叔丁基二甲基硅基(TBDMS)或叔丁基二苯基硅基(TBDPS)；更优选三甲基硅基(TMS)。
在本申请的一些实施方案中，所述式6化合物选自：
Figure PCTCN2018097158-appb-000012
在本申请的一些实施方案中，所述式7化合物选自：
Figure PCTCN2018097158-appb-000013
在本申请的一些实施方案中，所述式8化合物选自：
Figure PCTCN2018097158-appb-000014
在本申请的一些实施方案中，所述式10化合物选自：
Figure PCTCN2018097158-appb-000015
在本申请的一些实施方案中，所述式11化合物选自：
Figure PCTCN2018097158-appb-000016
定义和术语
除非另有说明，本文所用的术语和短语具有下文所列的含义。特定的术语或短语在没有特别定义的情况下不应该认为是不确定的或不清楚的，而应该按照本领域技术人员通常理解的含义进行解释。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。
在本申请中，所述的烷基包括其所有的同分异构体形式，例如，丙基包括正丙基、异丙基；丁基包括1-正丁基、2-正丁基、异丁基、叔丁基。
在本申请中，所述保护基及其连接或脱除方法可以采用本领域的常规方法实现，该方法可以是一步反应，也可以是多步反应，例如但不限于参照Wiley出版社的《Greene's Protective Groups in Organic Synthesis-4th Edition》或化学工业出版社的《保护基化学》实现。
在本申请中，化合物可以存在特定的几何或立体异构体形式。本申请设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体过量的混合物，所有这些混合物都属于本申请的范围之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。
在本申请中，所述pKa是酸度系数，代表酸离解氢离子的能力。按照本领域的通常理解，碱的pKa数值由其共轭酸所测得。
在本申请中，可以依据本领域的通常知识对甾体化合物的原子进行编号，例如但不限于参照邢其毅主编，高等教育出版社的《基础有机化学-第三版》。本申请中，以式7化合物为例，标注3位、7位和23位原子：
Figure PCTCN2018097158-appb-000017
有益效果
本申请的制备方法的反应条件温和，例如可以在常压下进行，反应温度易于控制。而且本申请所述的制备方法中部分步骤可以同时转变多个基团，有效缩短步骤，适合工业化生产。
具体实施方式
为了更好的理解本发明的内容，下面结合具体实施例来做进一步的说明，但具体的实施方式并不是对本发明的内容所做的限制。
制备实施例1：式8a化合物的制备
步骤1-1式5a化合物的制备
Figure PCTCN2018097158-appb-000018
在25摄氏度下，将甲醇(33L)加到50L反应釜中，将底物4(3.330kg，8.23mol)加到反应釜中，然后加入对甲苯磺酸一水合物(156.6g，0.823mol)，将反应加热到60摄氏度，在该温度下搅拌12小时。TLC监测反应，显示原料消失。HPLC显示约100％产物生成。反应液冷却至室温，然后用饱和的碳酸氢钠溶液调节pH值至9左右，旋干溶液得到粗品，粗品再用乙酸乙酯(30L)溶解，依次用饱和碳酸氢钠溶液(9L)、水(9L)、饱和食盐水(9L)洗涤。有机相旋干得到产品。产物性状为棕色油状液体。
1H NMR(400MHz,CDCl 3)δ3.66(s,3H),3.61-3.49(m,1H),2.74-2.66(m,1H),2.48-2.33(m,2H),2.24-2.15(m,1H),2.07-1.61(m,13H),1.54-1.40(m,3H),1.31-1.07(m,6H),1.02-0.77(m,9H),0.69(s,3H)。
步骤1-2式6a化合物的制备
Figure PCTCN2018097158-appb-000019
将式5a化合物(3100g)用二氯甲烷(30L)溶解，依次加入咪唑(529.4g)、三乙胺(786.8g)，在釜中降温(内温为5摄氏度)，在此温度下慢慢滴加TBDPSCl(2140g)，滴加过程中温度不超过10摄氏度，滴加完毕后反应在室温下搅拌反应16小时。TLC显示原料反应完全，在反应液中慢慢滴加15L水，淬灭反应。静置，反应分层，分出下层二氯甲烷相，将二氯甲烷相用饱和食盐水(10L)洗涤，浓缩有机相至成为产物。产物性状为棕色油状液体。
1H NMR(400MHz,CDCl 3)δ7.69-7.63(m,4H),7.45-7.34(m,6H),3.77(br t,J＝6.1Hz,1H),3.69(s,3H),3.54-3.44(m,1H),2.57(q,J＝6.1Hz,1H),2.46(br dd,J＝3.0,14.6Hz,1H),2.36-2.21(m,2H),2.08- 1.67(m,9H),1.62-1.17(m,12H),1.12-0.87(m,14H),0.70-0.62(m,6H)。
步骤1-3式7a化合物的制备
Figure PCTCN2018097158-appb-000020
在15℃下，将四氢呋喃(10L)加到50L反应釜中，在N 2保护下将LiAlH 4(235g，6.2mol)加到反应釜中，反应降温至内温为5℃。用四氢呋喃溶解式6a化合物(2.04kg)后，将其慢慢滴加到LiAlH 4的四氢呋喃溶液中，大约滴加2.5小时。反应在15℃搅拌2小时，TLC监测反应，显示原料消失。用H 2O(235mL)慢慢滴加到反应液中淬灭反应，再加入四氢呋喃溶液(20L)到反应液中，再将15％NaOH溶液(235mL)慢慢滴加到反应液中，搅拌12小时。将反应液过滤，二氯甲烷(3L)洗涤滤饼，滤液旋干，得到油状物。将油状物用DCM(15L)溶解后，有机相用水(5L)洗涤一次，饱和食盐水(5L)洗涤一次，滤液旋干，得到白色固体(1.8kg)，反应液冷却至室温(约16摄氏度)，然后用饱和的碳酸氢钠溶液调节pH值至9左右，旋干溶液(少量剩余)得到粗品，粗品用乙酸乙酯(30L)溶解，依次用饱和碳酸氢钠溶液(9L)、水(9L)、饱和食盐水(9L)洗涤。有机相旋干得到产品。产物性状为棕色油状液体。
1H NMR(400MHz,CDCl 3)δ7.64-7.58(m,4H),7.37-7.25(m,6H),3.68-3.52(m,3H),3.38-3.28(m,1H),1.91-1.03(m,25H),1.02-0.93(m,11H),0.88(d,J＝6.5Hz,3H),0.72-0.64(m,6H),0.57(s,3H)。
步骤1-4式8a化合物的制备
Figure PCTCN2018097158-appb-000021
向式7a化合物(3.52千克，5.58摩尔)的无水二氯甲烷(35升)溶液中加咪唑(1.14千克，16.73摩尔)。5℃，向体系中滴加三甲基氯硅烷(1770毫升，13.95摩尔)，两小时滴毕。体系于15℃搅拌3小时。TLC检测反应几乎完全。15℃向体系中加入10升水，搅拌，分液。有机相依次用10升水和10升饱和食盐水洗一次。
有机相浓缩至约5升，向溶液中加入30升乙醇。15℃，向溶液中加入碳酸钾(1.93千克，13.95摩尔)。体系于15℃搅拌14小时。TLC检测反应几乎完全。反应液过滤。用3升二氯甲烷淋洗滤饼。滤液浓缩，得到油状物。油状物用20升二氯甲烷溶解，依次用10升水和10升饱和食盐水洗一次。有机相用3千克无水硫酸钠干燥，过滤。硅胶柱层析(100-200目，230毫米×800毫米)纯化，正庚烷：乙酸乙酯＝30:1-20:1洗脱。得到目标化合物8a(3.20千克，82％产率，87％纯度)。
1H NMR(400MHz,CDCl 3)：δ7.77-7.64(m,4H),7.45-7.32(m,6H),3.78-3.56(m,3H),3.43-3.31(m,1H),1.98-1.13(m,24H),1.07(s,9H),0.97(d,J＝6.5Hz,3H),0.83-0.74(m,4H),0.68-0.55(m,6H),0.17-0.05(m,9H)。
制备实施例2：式I-Cl化合物的制备
步骤2-1式10a化合物的制备
Figure PCTCN2018097158-appb-000022
向反应釜中加入式8a化合物(2498.0g，3.10mol)，用THF(12.5L)溶解，控制内温5℃至10℃，t-BuONa(614.2g，6.20mol)缓慢加入，约40min加完，搅拌10min，升温至20℃至23℃下搅拌1.5h，降温至5℃至10℃。保持内温，向反应液中滴加式9a化合物的THF溶液(12.5L，6.20mol，1073.1g)，升温至60℃。搅拌1.5h，TLC和HPLC检测，反应完全，降温至20℃。加入25L水淬灭，乙酸乙酯萃取(25L×2)，合并有机相，饱和食盐水(25L×3)洗涤3次。旋干得油状粗品。粗品溶于2.5L丙酮，三个10L三口瓶中共加入6.6×3L甲醇。控制内温-10℃至-15℃，缓慢滴入粗品溶液，搅拌，析出大量固体。过滤，滤饼用3.0L甲醇洗涤，得到黄色固体(未干燥)黄色固体加入18.0L甲醇，打浆过夜。过滤，滤饼用3.0L甲醇洗涤，得黄色固体(未干燥)，黄色固体加入18.0L甲醇，打浆过夜过滤，滤饼用2.0L甲醇洗涤。真空干燥24h，得2522.0g黄色固体，即，式10a化合物(2522.0g，90％产率，纯度92.9％)。
1H NMR(400MHz,CHLOROFORM-d)δ＝8.25(d,J＝2.0Hz,1H),7.73(d,J＝2.0Hz,1H),7.63-7.51(m,4H),7.33-7.21(m,6H),4.48-4.27(m,2H),3.50(s,1H),3.31-3.18(m,1H),1.98-1.03(m,27H),0.95(s,9H),0.73-0.64(m,4H),0.58-0.46(m,6H),0.00(s,9H)。
步骤2-2式11a化合物的制备
Figure PCTCN2018097158-appb-000023
在反应釜(20L)加入式10a化合物(2520.0g，2.79mol)，加入EtOH(13.0L)，搅拌溶解，控制内温～10℃，加入NaOH(2232.0g，55.8mol)的水溶液(13.0L)，分批加入。升高温度至105℃，搅拌2.8小时。TLC和HPLC检测，反应完全。反应液冷却至10℃，静置两小时，瓶底析出固体。将上清液抽去19.5L向反应液中加入39.0L水，控制内温12摄氏度，搅拌36h。过滤，固体依次用6.0L水洗涤，6.0L乙腈洗涤。固体用10.0L乙腈打浆2小时，过滤，得固体。用12.0L丙酮打浆16小时。过滤，得固体。用12.0L丙酮再次打浆16小时。过滤，产品干燥，得到2332.3g白色固体式11a化合物(2332.3g，94.7％产率，纯度99.7％)。
1H NMR(400MHz,METHANOL-d 4)δ＝8.48(d,J＝2.0Hz,1H),8.07(d,J＝2.0Hz,1H),7.55(br dd,J＝6.5,12.5Hz,4H),7.41-7.11(m,6H),4.52-4.15(m,2H),3.54(br s,1H),3.34-3.22(m,1H),2.04-1.14(m,28H),0.93(s,9H),0.69(s,4H),0.60-0.43(m,6H),0.00(s,9H)。
步骤2-3式I-Cl化合物的制备
Figure PCTCN2018097158-appb-000024
在反应釜(50L)加入式11a化合物(2330.3g，2.65mmol)，加入THF(24.0L)溶解。控制内温10℃，缓慢滴加浓HCl(10.0L，120.00mol)液体经2h升温至13℃(室温)下搅拌90小时。TLC检测，在8℃至10℃下缓慢滴加75L氢氧化钠溶液(6000g)调pH至10，搅拌半小时用甲基叔丁基醚萃取(30L×4)，浓HCl(3000mL)调pH至5，乙酸乙酯萃取(30L×2)，有机相水洗(30L×4)，浓缩有机相，得到固体1350g。将固体用2.0L乙酸乙酯和5.0L正庚烷混合溶剂打浆过夜，过滤，得到固体1280g。用9.0L乙酸乙酯溶清后(80℃)，缓慢冷却至室温(10℃)。过滤，得到目标化合物1222g。
1H NMR(400MHz,METHANOL-d 4)δ＝8.69(d,J＝2.0Hz,1H),8.23(d,J＝2.0Hz,1H),4.67-4.30(m,2H),3.67(br s,1H),3.34-3.22(m,1H),2.10-1.11(m,25H),1.09-0.97(m,3H),0.96-0.86(m,6H),0.73(s,3H)。
将下述化合物进行药理或药代动力学评价：
Figure PCTCN2018097158-appb-000025
实验例1：体外评价
FXR生化实验
实验目的：
通过均相邻近发光放大实验(alphascreen)检测化合物对FXR结合反应的激活作用。
实验材料:
1.蛋白：谷胱甘肽-S-转移酶标记的FXR人源蛋白(Invitrogen)
2.共激活因子：生物素标记的类固醇受体辅活化子(Anaspec)
3.检测试剂：均相邻近发光放大实验(alphascreen)检测试剂盒(PerkinElmer)
实验方法：
1.化合物稀释：将待测化合物制备为40μM的DMSO溶液，随后将化合物3倍稀释至10个浓度点。参照化合物制备为400μM的DMSO溶液，随后以1.5倍稀释至10个浓度点。将稀释好的DMSO溶液以每孔150nL的体积加入384孔板的微孔中。
2.将谷胱甘肽-S-转移酶标记的FXR人源蛋白和生物素标记的类固醇受体辅活化子配制成浓度分别为0.4nM和30nM的混合溶液。以每孔15μL的体积加入384孔板的微孔中。室温孵育1小时。
4.将均相邻近发光放大实验(alphascreen)检测试剂盒中的受体小球混合液稀释125倍，以每孔7.5μL体积加入384孔板的微孔中。实验过程避光操作。室温孵育1小时。
5.将均相邻近发光放大实验(alphascreen)检测试剂盒中的供体小球混合液稀释125倍，以每孔7.5μL体积加入384孔板的微孔中。实验过程避光操作。室温孵育1小时。
6.EC 50测试：采用Envision于680nm波长处激发，读取520-620nm处的吸收信号。
7.分析数据:用Prism 5.0来分析数据，计算化合物的激活作用EC 50值。再将化合物的最高信号值与参照化合物的最高信号值作比值得出化合物的激活效力百分数(Efficacy)。
FXR细胞实验
实验目的：
通过β-内酰胺酶报告基因技术检测化合物对细胞功能活性的影响。
实验材料:
1.细胞系：FXR HEK 293T DA
2.细胞培养基：DMEM培养基添加10％血清和Penicillin/Streptomycin(1×)
3.检测试剂：
Figure PCTCN2018097158-appb-000026
报告基因检测试剂盒(Invitrogen)
实验方法：
1.化合物稀释：将待测化合物制备为100μM的DMSO溶液，随后将化合物3倍稀释至10个浓度点。参照化合物制备为100μM的DMSO溶液，随后以1.3倍稀释至10个浓度点。将稀释好的DMSO溶液以每孔200nL的体积加入384孔板的微孔中。
2.细胞接种：将FXR HEK 293T DA细胞复苏，用培养基重悬，稀释至密度为5×10 5个/mL，以每孔40μL的体积加入384孔板的微孔中。
3.将384微孔板于37℃、5％CO 2条件下培养16小时。
4.将6μL的1mM LiveBLAzer TM-FRET B/G(CCF4-AM)底物与60μL的B溶液和934μL的C溶液混合，以每孔8μL的体积加入384孔板的微孔中。
5.将384微孔板室温避光孵育2小时。
6.EC 50测试：采用Envision于409nm波长处激发，读取460nm和530nm处的吸收信号。
7.分析数据:用Prism 5.0来分析数据，计算化合物的激活作用EC 50值。再将测试化合物的最高信号值与参照化合物(鹅去氧胆酸，CDCA)的最高信号值作比值得出化合物的激活效力百分数(Efficacy)。
表1 检测生化实验测试结果以及细胞实验测试结果EC 50
供试样品 FXR酶活性 FXR细胞活性
EC 50(μM) Efficacy EC 50(μM) Efficacy
鹅去氧胆酸，CDCA 12.14 100％ 10.22 100％
式I-H化合物 0.006 249％
式I-Cl化合物 0.0025 248％ 0.003 150％
式I-F化合物 0.0025 138％
结论：本申请化合物对FXR受体的激动作用显著，在细胞水平上对FXR受体的激动作用也比较显著。
实验例2：体内研究
单独给药小鼠药代：
12只C57BL/6J雄性小鼠随机分为两组，6只每组。第一组为静脉组，为尾静脉脉注射给药2mg/kg、2mL/kg(溶媒为10％HPbCD水溶液，药物溶解度不理想时，则会加入助溶剂)；第二组为口服组，灌胃给药10mg/kg、10mL/kg(溶媒为0.5％HPMC水溶液)。静脉组给药后0.083、0.25、0.5、1、2、4、6、8和24小时采集血浆(K 2-EDTA为抗凝)样品；口服组给药后0.25、0.5、1、2、4、6、8和24小时采集血浆样品。每组6只动物，一个时间点采集3个动物血样，第1批3只动物与第2批3只动物交错采样。使用LC-MS/MS进行血浆样品分析。获得血浆浓度与时间作图，并使用Phoenix WinNonlin 6.3计算PK参数。
表2
Figure PCTCN2018097158-appb-000027
结论：如表2中所示，相同剂量口服给药后式I-H化合物的峰值浓度与对照化合物obeticholic acid接近，药物暴露量高于对照化合物obeticholic acid。相同剂量口服给药后式I-Cl化合物的峰值浓度高于对照化合物obeticholic acid，药物暴露量也高于对照化合物obeticholic acid。
盒式给药小鼠肝血比实验：
6只C57BL/6J雄性小鼠为一组，为口服组，制剂中含有5种研发药物，灌胃给药2mg/kg/化合物(溶媒为0.5％HPMC水溶液)。5个化合物首先分别溶于溶媒中，通过超声或者涡旋，分别形成1mg/mL溶液(澄清溶液或者混悬液)，然后五种化合物溶液等体积混合(1:1:1:1:1，v:v:v:v:v)于一个玻璃瓶中。灌胃口服给药后，3只动物于给药后0.5小时采集血浆和肝组织样品；另外3只动物于给药后3小时采集相应样品。肝组织采集后，使用冰冷匀浆缓冲液(甲醇:15mM PBS缓冲液(pH 7.4)＝1:2，v:v)按照肝重量:匀浆缓冲液体积＝1:3进行匀浆。使用提前开发的五合一的LC-MS/MS分析方法进行血浆和肝组织样品分析。获得血浆浓度以及肝组织匀浆液浓度，并使用Excel计算肝组织与血浆浓度比值。
表3
Figure PCTCN2018097158-appb-000028
结论：如表3中所示，口服给予相同剂量本申请化合物后，式I-H化合物在0.5小时以及3小时于肝脏中的药物浓度均高于对照化合物，0.5小时以及3小时肝/血浓度比也高于对照化合物。

Claims (20)

  1. 一种制备式I化合物的方法，
    Figure PCTCN2018097158-appb-100001
    包括：
    步骤a)使式8化合物和式9化合物反应以获得式10化合物，
    Figure PCTCN2018097158-appb-100002
    步骤b)从式10化合物反应以获得式11化合物，
    Figure PCTCN2018097158-appb-100003
    步骤c)从式11化合物反应以获得式I化合物，
    Figure PCTCN2018097158-appb-100004
    其中，R 1选自H、Cl、Br或F，
    P 1和P 2各自独立地选自羟基保护基，
    M选自金属阳离子。
  2. 如权利要求1所述的制备方法，其中，所述P 1和P 2各自独立地选自在碱性条件下与上述甾体化合物3位或/和7位所连氧原子保持连接的羟基保护基；优选地，所述P 1和P 2各自独立地选自在pH≥7.5的条件下与上述甾体化合物3位或/和7位所连氧原子保持连接的羟基保护基。
  3. 如权利要求1所述的制备方法，所述P 1选自硅烷醚类保护基、烷基醚类保护基、烷氧基甲基醚类保护基或酯类保护基；优选地，所述P 1选自三甲基硅基、三乙基硅基、叔丁基二甲基硅基、叔丁基二苯基硅基、苄基、对甲氧基苄基、3,4-二甲氧基苄基、甲基醚、叔丁基、三苯甲基、4,4’-二甲氧基三苯甲基、甲氧基甲基、苄氧基甲基、四氢呋喃基、甲酰基、乙酰基、氯乙酰基、甲氧基乙酰基或新戊酰基。
  4. 如权利要求1所述的制备方法，所述P 2选自硅烷醚类保护基；优选地，所述P 2选自三甲基硅基、三乙基硅基、叔丁基二甲基硅基或叔丁基二苯基硅基。
  5. 如权利要求1所述的制备方法，所述M选自碱金属阳离子或碱土金属阳离子；优选地，所述M选自Li +、Na +、K +、Ca 2+或Mg 2+。
  6. 如权利要求1所述的制备方法，在步骤a)中，反应在碱的存在下进行；优选地，所述碱选自pKa≥12的碱。
  7. 如权利要求1所述的制备方法，在步骤b)中，反应在碱的存在下进行；优选地，所述碱选自pKa≥12的碱。
  8. 如权利要求1所述的制备方法，在步骤c)中，反应在酸的存在下进行；优选地，所述酸使得反应在pH≤6的条件下进行。
  9. 如权利要求1-8中任一项所述的制备方法，其还包括：
    步骤d)以式4化合物为原料，与醇发生酯化反应以获得式5化合物，
    Figure PCTCN2018097158-appb-100005
    步骤e)式5化合物3位羟基连接保护基以获得式6化合物，
    Figure PCTCN2018097158-appb-100006
    步骤f)从式6化合物反应以获得式7化合物，
    Figure PCTCN2018097158-appb-100007
    步骤g-1)，式7化合物7位和23位羟基连接保护基P 2以获得式12化合物，步骤g-2)，从式12化合物反应脱除23位羟基保护基P 2以获得式8化合物，
    Figure PCTCN2018097158-appb-100008
    其中，R 2选自C 1-6烷基。
  10. 如权利要求9所述的制备方法，所述R 2选自C 1-4烷基，优选甲基、乙基、丙基或丁基。
  11. 一种制备式8化合物的制备方法，包括如下步骤：
    步骤d)以式4化合物为原料，与醇发生酯化反应以获得式5化合物，
    Figure PCTCN2018097158-appb-100009
    步骤e)式5化合物3位羟基连接保护基以获得式6化合物，
    Figure PCTCN2018097158-appb-100010
    步骤f)从式6化合物反应以获得式7化合物，
    Figure PCTCN2018097158-appb-100011
    步骤g-1)，式7化合物7位和23位羟基连接保护基P 2以获得式12化合物，步骤g-2)从式12化合物反应脱除23位羟基保护基P 2以获得式8化合物，
    Figure PCTCN2018097158-appb-100012
    其中，P 1和P 2各自独立地选自羟基保护基，
    R 2选自C 1-6烷基。
  12. 如权利要求11所述的制备方法，其中，所述P 1和P 2各自独立地选自在非酸性条件下可有效保护上述甾体化合物3位或/和7位羟基的保护基；优选地，所述P 1和P 2各自独立地选自在pH≥7的条件下与上述甾体化合物3位或/和7位所连氧原子保持连接的羟基保护基。
  13. 如权利要求11所述的制备方法，所述P 1选自硅烷醚类保护基、烷基醚类保护基、烷氧基甲基醚类保护基或酯类保护基；优选地，所述P 1选自三甲基硅基、三乙基硅基、叔丁基二甲基硅基、叔丁基二苯基硅基、苄基、对甲氧基苄基、3,4-二甲氧基苄基、甲基醚、叔丁基、三苯甲基、4,4’-二甲氧基三苯甲基、甲氧基甲基、苄氧基甲基、四氢呋喃基、甲酰基、乙酰基、氯乙酰基、甲氧基乙酰基或新戊酰基。
  14. 如权利要求11所述的制备方法，所述P 2选自硅烷醚类保护基；优选地，所述P 2选自三甲基硅基、三乙基硅基、叔丁基二甲基硅基或叔丁基二苯基硅基。
  15. 如权利要求11-14中任一项所述的制备方法，所述R 2选自C 1-4烷基，优选甲基、乙基、丙基或丁基。
  16. 如下所示化合物：
    Figure PCTCN2018097158-appb-100013
    其中，R 1选自H、Cl、Br或F，
    R 2选自甲基、乙基、丙基或丁基，
    P 1选自硅烷醚类保护基、烷基醚类保护基、烷氧基甲基醚类保护基或酯类保护基，
    P 2选自硅烷醚类保护基，
    M选自金属阳离子。
  17. 如权利要求16所述的化合物，所述P 1选自三甲基硅基、三乙基硅基、叔丁基二甲基硅基、叔丁基二苯基硅基、苄基、对甲氧基苄基、3,4-二甲氧基苄基、甲基醚、叔丁基、三苯甲基、4,4’-二甲氧基三苯甲基、甲氧基甲基、苄氧基甲基、四氢呋喃基、甲酰基、乙酰基、氯乙酰基、甲氧基乙酰基或新戊酰基。
  18. 如权利要求16所述的化合物，所述P 2选自三甲基硅基、三乙基硅基、叔丁基二甲基硅基或叔丁基二苯基硅基。
  19. 如权利要求16所述的化合物，所述M选自碱金属阳离子或碱土金属阳离子；优选Li +、Na +、K +、Ca 2+或Mg 2+；更优选Na +或K +。
  20. 如权利要求16所述的化合物，选自：
    Figure PCTCN2018097158-appb-100014
    Figure PCTCN2018097158-appb-100015
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2020528067A (ja) * 2017-07-26 2020-09-17 チア タイ ティエンチン ファーマシューティカル グループ カンパニー リミテッドChia Tai Tianqing Pharmaceutical Group Co., Ltd. ステロイド系誘導体ｆｘｒアゴニストの結晶又は非晶質、その製造方法及び使用

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000037077A1 (en) 1998-12-23 2000-06-29 Glaxo Group Limited Assays for ligands for nuclear receptors
CN105102425A (zh) * 2012-10-26 2015-11-25 英特塞普特医药品公司 制备胆汁酸衍生物的方法
WO2016161003A1 (en) * 2015-03-31 2016-10-06 Enanta Phamraceuticals, Inc. Bile acid derivatives as fxr/tgr5 agonists and methods of use thereof
CN106083978A (zh) * 2015-04-28 2016-11-09 上海迪诺医药科技有限公司 磺酰基氨基羰基衍生物、其药物组合物及应用
CN106661079A (zh) * 2014-05-29 2017-05-10 巴尔制药有限公司 用于治疗和/或预防fxr和tgr5/gpbar1介导的疾病的胆烷衍生物
WO2017129125A1 (zh) * 2016-01-28 2017-08-03 正大天晴药业集团股份有限公司 甾体类衍生物fxr激动剂

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107531743B (zh) * 2015-04-29 2020-05-05 正大天晴药业集团股份有限公司 鹅去氧胆酸衍生物
EP3331896A4 (en) 2015-08-07 2019-08-14 Intercept Pharmaceuticals, Inc. PROCESSES FOR PREPARING BILIARY ACIDS AND THEIR DERIVATIVES
EP3352768B1 (en) 2015-09-21 2022-11-02 Intercept Pharmaceuticals, Inc. Methods of promoting hepatic regeneration
AU2016325619A1 (en) * 2015-09-24 2018-04-12 Intercept Pharmaceuticals, Inc. Methods and intermediates for the preparation bile acid derivatives
MX2018005520A (es) 2015-11-06 2018-08-01 Intercept Pharmaceuticals Inc Metodos para preparacion de acido obeticolico y derivados de los mismos.
US10323060B2 (en) * 2016-02-23 2019-06-18 Enanta Pharmaceuticals, Inc. Benzoic acid derivatives of bile acid as FXR/TGR5 agonists and methods of use thereof
US11059853B2 (en) 2017-07-26 2021-07-13 Chia Tai Tianqing Pharmaceutical Group Co., Ltd. Crystalline or amorphous form of steroid derivative FXR agonist, preparation method therefor and use thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000037077A1 (en) 1998-12-23 2000-06-29 Glaxo Group Limited Assays for ligands for nuclear receptors
CN105102425A (zh) * 2012-10-26 2015-11-25 英特塞普特医药品公司 制备胆汁酸衍生物的方法
CN106661079A (zh) * 2014-05-29 2017-05-10 巴尔制药有限公司 用于治疗和/或预防fxr和tgr5/gpbar1介导的疾病的胆烷衍生物
WO2016161003A1 (en) * 2015-03-31 2016-10-06 Enanta Phamraceuticals, Inc. Bile acid derivatives as fxr/tgr5 agonists and methods of use thereof
CN106083978A (zh) * 2015-04-28 2016-11-09 上海迪诺医药科技有限公司 磺酰基氨基羰基衍生物、其药物组合物及应用
WO2017129125A1 (zh) * 2016-01-28 2017-08-03 正大天晴药业集团股份有限公司 甾体类衍生物fxr激动剂

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
"Greene's Protective Groups in Organic Synthesis", WILEY PRESS, OR PROTECTING GROUPS PUBLISHED BY CHEMICAL INDUSTRY PRESS
DATABASE REGISTRY 23 June 2014 (2014-06-23), retrieved from STN Database accession no. RN 1612191-84-0 *
DJ. MANGELSDORF ET AL., CELL, vol. 83, 1995, pages 841 - 850
See also references of EP3660030A4
W. SEOL ET AL., MOL. ENDOCRINNOL, vol. 9, 1995, pages 72 - 85

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2020528067A (ja) * 2017-07-26 2020-09-17 チア タイ ティエンチン ファーマシューティカル グループ カンパニー リミテッドChia Tai Tianqing Pharmaceutical Group Co., Ltd. ステロイド系誘導体ｆｘｒアゴニストの結晶又は非晶質、その製造方法及び使用
EP3660031A4 (en) * 2017-07-26 2021-03-31 Chia Tai Tianqing Pharmaceutical Group Co., Ltd. CRYSTALLINE OR AMORPHIC FORM OF STEROID DERIVATIVE FXR AGONIST, METHOD OF MANUFACTURING THEREOF AND USE THEREOF
JP7244487B2 (ja) 2017-07-26 2023-03-22 チア タイ ティエンチン ファーマシューティカル グループ カンパニー リミテッド ステロイド系誘導体ｆｘｒアゴニストの結晶又は非晶質、その製造方法及び使用

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