CN116444597A - Method for preparing deoxycholic acid intermediate and deoxycholic acid - Google Patents

Method for preparing deoxycholic acid intermediate and deoxycholic acid Download PDF

Info

Publication number
CN116444597A
CN116444597A CN202310323916.XA CN202310323916A CN116444597A CN 116444597 A CN116444597 A CN 116444597A CN 202310323916 A CN202310323916 A CN 202310323916A CN 116444597 A CN116444597 A CN 116444597A
Authority
CN
China
Prior art keywords
compound
reaction
oxidation reaction
oxidizing agent
acid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202310323916.XA
Other languages
Chinese (zh)
Inventor
林成刚
周春燕
张翠霞
吴刚
赵欣
姜伟化
孙小燕
刘飞
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing Minoway Medical Technology Co ltd
Original Assignee
Nanjing Minoway Medical Technology Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nanjing Minoway Medical Technology Co ltd filed Critical Nanjing Minoway Medical Technology Co ltd
Priority to CN202310323916.XA priority Critical patent/CN116444597A/en
Publication of CN116444597A publication Critical patent/CN116444597A/en
Pending legal-status Critical Current

Links

Classifications

    • 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
    • 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
    • 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

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Steroid Compounds (AREA)

Abstract

The present invention provides a process for preparing compound N comprising: the compound M is subjected to oxidation reaction to obtain a compound N, wherein the reaction formula is shown as follows. Wherein: p is a hydroxyl protecting group; r is alkyl; in steroid ringsRepresents a single bond or a double bond; with or without substituents on the steroid ring; n represents the number of methylene groups, and n is a natural number of 1-5; the methylene group may have a substituent thereon; the oxidizing agent of the oxidation reaction is selected from the group consisting of iodobenzene diacetic acid, 2-iodoxybenzoic acid, and dessert-martin reagent. The method can be used for preparing key intermediates of deoxycholic acid, does not use chromium-containing oxidizing reagent, is environment-friendly, has low production cost and good yield, and is suitable for industrial application.

Description

Method for preparing deoxycholic acid intermediate and deoxycholic acid
Technical Field
The invention belongs to the field of pharmaceutical chemistry, and in particular relates to a method for preparing deoxycholic acid and an intermediate thereof.
Background
Deoxycholic acid (formula I) is a bile acid with a C-7 hydroxyl group, and is a free bile acid obtained by derivatization of cholic acid by losing an oxygen atom. Deoxycholic acid has stronger surface activity, and can accurately destroy the cell membrane of fat cells after local injection, so that the fat cells are broken and phagocytized by macrophages, and tissue cells such as skin, muscle and the like cannot be cracked because the cell membrane surface contains a large amount of protein, thereby reducing the dissolution of small-range local subcutaneous fat. In 2015, deoxycholic acid injection developed by Kythera was approved by the United states food and drug administration as a drug useful for injecting fat, and was marketed under the trade name Kybella for improving the protrusion or plumpness (double chin) of medium to severe submental fat in adults.
Deoxycholic acid is currently mainly derived from animal carcasses, although at low cost, with the risk of potentially containing animal pathogens and other deleterious factors. Methods for preparing deoxycholic acid by chemical synthesis have been disclosed in the prior art.
CN106083969a discloses a total synthesis method of deoxycholic acid, as shown in scheme 1 below. The method uses chromium trioxide as an oxidant in the synthesis of intermediate 1.10. The oxidant has high toxicity, high pollution and carcinogenicity.
Scheme 1
CN107011401 discloses a method for preparing deoxycholic acid, the synthetic route is shown in scheme 2. The method adopts an oxidation system of tert-butyl peroxide (TBHP) and sodium hypochlorite in the synthesis of intermediate 2.24, thereby avoiding the application of chromium trioxide. Pyridinium chlorochromate (PCC) was used in the preparation of compound 2.24; byproducts are generated in the preparation process of 2.33, and the yield is low. To increase the reaction yield, the by-product 2.34 needs to be further oxidized with pyridinium chlorochromate to give 2.33.
Scheme 2
CN106146593 discloses a method for preparing deoxycholic acid, the synthetic route is shown in scheme 3. In the preparation of intermediates 3.4 and 3.5, it is also desirable to use pyridinium chlorochromate as an oxidant to enhance the conversion of byproducts to the target intermediate to enhance the yields of these two steps.
Scheme 3
The chromium trioxide and the pyridinium chlorochromate used in the prior art are all metal-containing chromium oxidants. Chromium is one of the most toxic pollutants, has carcinogenic and mutagenic effects, has stimulation and corrosion effects on human respiratory tract, can cause rhinitis, tuberculosis, bronchitis and other diseases, and can also induce lung cancer, nasopharyngeal carcinoma and the like. The discharge of industrial chromium-containing waste gas and wastewater is a main source of artificial chromium pollution, for which, china issues a plurality of regulations on the prevention and treatment of environmental pollution in chromium compound production construction, industrial pollutant discharge standards of chromium and chromium compounds, notification on the reinforcement of the prevention and treatment of chromium-containing hazardous waste pollution, and chromium-containing waste is incorporated into the national hazardous waste directory, and strict index control standards are provided for chromium pollutant discharge.
Enterprises need to carry out complex and special treatment procedures when discharging chromium-containing waste gas and wastewater, for example, a method of extraction and washing is generally adopted to remove chromium-containing components, and a large amount of water resources are needed to be consumed; or the generated chromium-containing solid waste is treated by a chemical precipitation method, so that secondary pollution is caused, enterprises with hazardous solid waste disposal qualification are required to be entrusted to reprocess, the utilization value is low, the transportation and the storage are inconvenient, and high sludge disposal cost is required to be paid. Therefore, there is a need to develop a method that does not use chromium-containing oxidants, regardless of environmental protection and production costs.
Disclosure of Invention
The invention aims to overcome the problems and provide an oxidation method of steroid derivatives, in particular to a method for preparing deoxycholic acid intermediates, which does not need to use chromium-containing oxidants, is environment-friendly, low in production cost, high in yield and suitable for industrial application.
The present invention provides a process for preparing compound N comprising: the compound M is subjected to oxidation reaction to obtain a compound N, wherein the reaction formula is as follows:
wherein:
p is a hydroxy protecting group selected from benzyl, P-methoxybenzyl, benzoyl, acetyl, formyl and benzyloxycarbonyl; preferably, P is benzoyl or acetyl;
in steroid ringsRepresents a single bond or a double bond; preferably, in the oxidation reaction +.>The type of single double bond of (2) is not changed; optionally, the steroid ring may or may not have substituents thereon;
r is an alkyl group, preferably a C1-C4 alkyl group, such as methyl, ethyl, propyl, butyl, and the like.
n represents the number of methylene groups, n is a natural number of 1 to 5, for example 1, 2, 3, 4, 5; optionally, the methylene group may or may not have a substituent thereon, for example the substituent may be an alkyl group, preferably a C1-C4 alkyl group;
the oxidizing agent of the oxidation reaction is selected from the group consisting of iodobenzene diacetic acid, 2-iodoxybenzoic acid (IBX) and dessert-martin reagent (DMP). Preferably, the oxidizing agent is 2-iodoxybenzoic acid (IBX) or dessmartin reagent (DMP).
In some embodiments, compound M, N is represented by the following formulas M1, N1, respectively:
wherein, in the structural formula, P, R,Is defined as above.
In other embodiments, compound M, N is represented by the following formulas M2, N2, respectively:
wherein, in the structural formula, P, R,Is as defined above.
In other embodiments, compound M, N is represented by the following formulas M3, N3, respectively:
wherein, in the structural formula, P,Is as defined above.
In some embodiments, the molar ratio of oxidizing agent to compound M of the oxidation reaction may be (0.5-10): 1, preferably (0.8-5): 1, more preferably (1-3): 1.
In some embodiments, the reaction solvent of the oxidation reaction includes a dipolar aprotic solvent, such as dimethyl sulfoxide (DMSO), dimethylformamide (DMF), N-methylpyrrolidone (NMP), dimethylacetamide (DMA), acetonitrile, and the like.
In one exemplary embodiment, a process for preparing deoxycholic acid intermediate compound IV is provided, comprising subjecting compound IV-a to an oxidation reaction to obtain compound IV;
the reaction formula is as follows:
wherein P is a hydroxy protecting group, e.g., P is selected from benzyl, P-methoxybenzyl, benzoyl, acetyl, formyl and benzyloxycarbonyl; preferably, P is benzoyl or acetyl;
The oxidizing agent of the oxidation reaction is selected from the group consisting of 2-iodoxybenzoic acid and dess martin reagent.
In some embodiments, the molar ratio of compound IV-a to oxidizing agent may be (0.5-10): 1, preferably (0.8-5): 1, more preferably (1-3): 1.
In some embodiments, the reaction solvent of the oxidation reaction includes a dipolar aprotic solvent, such as dimethyl sulfoxide (DMSO), dimethylformamide (DMF), N-methylpyrrolidone (NMP), dimethylacetamide (DMA), acetonitrile, and the like.
In another exemplary embodiment, a process for preparing deoxycholic acid intermediate compound V is provided, comprising subjecting compound V-b to an oxidation reaction to obtain compound V;
the reaction formula is as follows:
p is a hydroxyl protecting group, for example, P is one selected from benzyl, P-methoxybenzyl, benzoyl, acetyl, formyl and benzyloxycarbonyl; more preferably, P is benzoyl or acetyl;
the oxidizing agent of the oxidation reaction is selected from the group consisting of 2-iodoxybenzoic acid and dess martin reagent.
In some embodiments, the molar ratio of oxidizing agent to compound V-b may be (0.5-10): 1, preferably (0.8-5): 1, more preferably (1-3): 1.
In some embodiments, the reaction solvent of the oxidation reaction includes a dipolar aprotic solvent, such as dimethyl sulfoxide (DMSO), dimethylformamide (DMF), N-methylpyrrolidone (NMP), dimethylacetamide (DMA), acetonitrile, and the like.
The invention further provides a method for preparing deoxycholic acid, comprising the following steps:
1) The compound II is subjected to hydrogenation reaction to obtain a compound III;
2) Carrying out a first oxidation reaction on the compound III to obtain a mixture containing the compound IV and the compound IV-a; the mixture is subjected to a second oxidation reaction to obtain a compound IV;
3) The compound IV is subjected to a first hydrogenation reaction to obtain a mixture containing compounds V, V-a and V-b; the mixture sequentially undergoes oxidation reaction and second hydrogenation reaction to obtain a compound V;
4) The compound V is subjected to reduction reaction to obtain a compound VI;
5) Deprotection and hydrolysis reaction of the compound VI to obtain deoxycholic acid I or salt thereof;
the reaction formula is as follows:
wherein P is a hydroxyl protecting group; can be selected from benzyl, p-methoxybenzyl, acetyl, benzoyl, formyl and benzyloxycarbonyl; preferably, P is benzoyl or acetyl.
In some embodiments, in step 1), the hydrogenation is carried out in Pd/C or PtO 2 In the presence of a catalyst; pd/C catalysts are preferred.
In some embodiments, in step 2), the first oxidation reaction is performed in the presence of an oxidizing agent and a co-oxidizing agent. The oxidizing agent is preferably t-butyl hydroperoxide (TBHP). The co-oxidant is selected from aqueous hypochlorous acid solution, pd/C, pd (OCOCF) 3 ) 2 、Pd(OAc) 2 And CuI; preferably aqueous hypochlorous acid or CuI; more preferably CuI.
In some embodiments, in step 2), the oxidizing agent used in the second oxidation reaction is selected from the group consisting of 2-iodoxybenzoic acid and dessert-martin reagent.
In some embodiments, in step 2), the molar ratio of oxidizing agent to compound IV-a used in the second oxidation reaction may be (0.8-5): 1, e.g., (1-3): 1, e.g., 1:1, 1.5:1, 2:1, etc.
In some embodiments, in step 2), the reaction solvent of the second oxidation reaction comprises a dipolar aprotic solvent, such as dimethyl sulfoxide (DMSO), dimethylformamide (DMF), N-methylpyrrolidone (NMP), dimethylacetamide (DMA), acetonitrile, and the like.
In some embodiments, in step 3), compound IV is subjected to a first hydrogenation reaction to obtain a mixture comprising compounds V, V-a and V-b; the mixture containing the compounds V, V-a and V-b is subjected to oxidation reaction to obtain a mixture basically containing the compounds IV and V; and (3) carrying out a second hydrogenation reaction on the mixture basically containing the compounds IV and V to obtain a compound V.
In some embodiments, in step 3), the first hydrogenation reaction and the second hydrogenation reaction are carried out at Pd/C or PtO 2 In the presence of a catalyst; pd/C catalysts are preferred.
In some embodiments, in step 3), the oxidizing agent used in the oxidation reaction is selected from the group consisting of 2-iodoxybenzoic acid and dessert-martin reagent.
In some embodiments, in step 3), the reaction solvent of the oxidation reaction comprises a dipolar aprotic solvent, such as dimethyl sulfoxide (DMSO), dimethylformamide (DMF), N-methylpyrrolidone (NMP), dimethylacetamide (DMA), acetonitrile, and the like.
In some embodiments, in step 3), the oxidizing agent is oxidized with both compounds V-a and V-b. The molar ratio of oxidizing agent to the sum of the compounds V-a and V-b is from (0.5 to 10): 1, preferably from (0.8 to 5): 1, for example from (1 to 3): 1.
In some embodiments, in step 4), the reducing agent used in the reduction reaction is lithium aluminum tri-tert-butoxide (LiAl (OtBu) 3 )。
In some embodiments, in step 5), the deprotection and hydrolysis reactions may be performed according to known methods. For example, the deprotection and hydrolysis conditions comprise reacting compound VI with an alkaline earth metal hydroxide, an alkaline earth metal alkoxide or a mixture thereof. In certain embodiments, the hydrolysis conditions include an acid treatment to obtain deoxycholic acid. In certain embodiments, the acid treatment is omitted to obtain the corresponding salt. The alkaline earth metal hydroxide may be lithium hydroxide (LiOH) or sodium hydroxide (NaOH).
The method of the invention attempts to efficiently complete the oxidation reaction by adopting new oxidants (such as IBX and DMP), avoids the harm to the environment and man-made by using chromium-containing oxidants, does not need to spend a great deal of cost for treating chromium-containing waste liquid, and remarkably reduces the production cost. The method can be used for preparing the intermediate compound IV and the intermediate compound V of deoxycholic acid, is environment-friendly, convenient in post-treatment, low in cost and high in yield, and is suitable for industrial production.
Detailed Description
The present invention will be described in detail with reference to examples. It should be understood that the methods in the examples are for illustrative purposes only and are not to be construed as limiting the invention in any way. The materials used in the examples are commercially available unless otherwise specified. Unless otherwise indicated, all experimental procedures used were conventional. In the examples, the completion of the reaction was monitored by HPLC and the mass percent of the compound was determined using area normalization. Unless otherwise indicated, the percentages of solutions used in the examples are all percentages by mass of solutes contained.
The number of the steroid structure skeleton in the invention follows the general rule:
unless otherwise indicated, the backbone represents only the position of carbon atoms. The substituents on the backbone of the support structure may include the R configuration, the S configuration, or mixtures thereof, unless specifically indicated.
The Pd/C catalyst in the invention refers to a catalyst with Pd loaded on C, and can be used for catalyzing hydrogenation reaction. The "Pd/C" catalyst may be free of water or contain some amount of water. Pd/C catalysts of various sizes (e.g., 5% Pd, 10% Pd), etc., can be used in the present invention. The catalyst may be added in conventional amounts.
In the present invention, the "hydroxyl protecting group P" means a protecting group well known in the art of organic synthesis as being suitable for preventing side reactions of hydroxyl groups. For example, many protecting groups are set forth in T.W. Green (T.W. Greene) and G.M. Wu Ci (G.M.Wuts), protecting groups in organic synthesis (Protecting Groupsin Organic Synthesis), 3 rd edition, wiley Press (Wiley), new York, 1999 and references cited therein. Representative hydroxyl protecting groups include, but are not limited to: alkyl groups (such as methyl, ethyl, and t-butyl); acyl (e.g., formyl, acetyl, benzoyl); arylmethyl (e.g., benzyl Bn, p-methoxybenzyl PMB, 9-fluorenylmethyl Fm, diphenylmethyl DPM; silyl (e.g., trimethylsilyl TMS, t-butyldimethylsilyl TBS), etc. preferably, the hydroxy protecting group is acetyl or benzoyl.
"alkyl" according to the invention means a saturated monovalent hydrocarbon radical, preferably a C1-C4 alkyl radical, such as methyl, ethyl, propyl, isopropyl, butyl, etc.
The intermediate compound IV of deoxycholic acid in the invention can be prepared by taking a compound II or a compound III as a starting material. Methods for the preparation of compounds II, III are disclosed, for example, in patent applications CN106146593A, CN101711254A, CN107011401A, CN102512035A, CN103906517A or CN114805463A, etc. As an illustrative example, compound IIA in the following examples was prepared by the method of CN106146593 a. Compound IIB was prepared by the method described in connection with CN 101711254A.
Example 1: synthesis of Compound IIIA
After adding Compound IIA (50 g,0.1 mol), tetrahydrofuran (500 mL, 10V), and 10% Pd/C (50% wet,2.5g,5% wt) to a reaction vessel, the reaction was carried out sequentially with nitrogen substitution and hydrogen substitution, and the reaction was completed by HPLC monitoring at a temperature of 25.+ -. 10 ℃ and a hydrogen pressure of 1.2MPa for 24 hours (Compound IIA < 1%). After nitrogen substitution of the reaction vessel, the reaction solution was filtered, the solvent was removed from the filtrate under reduced pressure, acetonitrile (150 mL, 3V) was added to the solution, the temperature was lowered to-5.+ -. 5 ℃ and stirred for 6 hours, filtration was carried out, and washing with acetonitrile (25 mL, 0.5V) precooled to-15.+ -. 5 ℃ was carried out to obtain 47.2g of Compound IIIA, the yield was 94.0%, and the HPLC purity was 99.5%.
1 HNMR(400MHz,CDCl 3 )δ8.03(m,2H),7.51(t,J＝7.4Hz,1H),7.42(m,2H),5.36(m,1H),4.97(m,1H),3.67(s,3H),2.34-2.32(m,1H),2.23-2.21(m,1H),2.15-1.54(m,13H),1.54-1.06(m,10H),1.10(s,3H),0.94(d,J＝6.4Hz,3H),0.59(s,3H). 13 CNMR(400MHz,DMSO)δ174.2(1C,C 24 ),165.6(1C,C 26 ),140.2(1C,C 9 ),133.7(1C,C 30 ),130.5(1C,C 27 ),129.5(2C,C 28 ,C 32 ),129.1(2C,C 29 ,C 31 ),119.5(1C,C 11 ),75.2(1C,C 3 ),55.9(1C,C 17 ),53.2(1C,C 14 ),51.7(1C,C 25 ),41.9,41.5,40.9,38.7,36.4,35.2,35.1,33.9,31.0,30.8,29.8,28.2,28.0,27.0,26.7,25.3(1C,C 21 ),18.2(1C,C 19 ),12.0(1C,C 18 ) The mass spectrum (ES (+)) measures the [ M+NH ] of the product 4 ] + The mass to charge ratio of the peak was 510.6.
Example 2: synthesis of Compound IIIA
After adding compound IIA (50 g,0.1 mol), tetrahydrofuran (500 mL, 10V), and 10% Pd/C (50% wet.,2.5g,5% wt.) to the reaction vessel, the apparatus was sealed, and then subjected to nitrogen substitution and hydrogen substitution, and reacted at a temperature of 30.+ -. 10 ℃ and a hydrogen pressure of 1.3MPa for 16 hours. The reaction was determined to be complete by HPLC monitoring (compound iia < 1%). After nitrogen replacement of the reaction vessel, the reaction solution was filtered, the solvent was removed from the filtrate under reduced pressure, acetonitrile (150 mL, 3V) was added to the filtrate, the temperature was lowered to-5.+ -. 5 ℃ and stirred for 5 hours, the mixture was filtered, and the mixture was washed with acetonitrile (25 mL, 0.5V) precooled to-15.+ -. 5 ℃ to obtain 47.8g of Compound IIIA, yield 95.2% and HPLC purity 99.3%.
EXAMPLE 3 Synthesis of Compound IIIA
To the reaction vessel, compound IIA (1.0 Kg,2.0 mol), tetrahydrofuran (5L, 5V) and 10% Pd/C (50% wet,120g,12% wt) were charged, followed by nitrogen substitution and hydrogen substitution in this order, and then reacted at 25.+ -. 10 ℃ under a hydrogen pressure of 1.0MPa for 24 hours. The reaction was determined to be complete by HPLC monitoring (compound iia < 1%). After the nitrogen gas replacement of the reaction vessel, the reaction solution was filtered, the solvent was removed from the filtrate under reduced pressure, acetonitrile (2L, 2V) was added, the solvent was removed by distillation under reduced pressure continuously, then acetonitrile (12L, 12V) was added to heat the supernatant, the temperature was lowered to-5.+ -. 5 ℃ and stirred for 5 hours, the mixture was filtered, and washed with acetonitrile (500 mL, 0.5V) precooled to-15.+ -. 5 ℃ to obtain a compound IIIA963.6g, yield 96.0% and HPLC purity 99.4%.
Example 4: synthesis of Compound IIIA
After adding Compound IIA (1.0 Kg,2.0 mol), tetrahydrofuran (10L, 10V) and 10% Pd/C (50% wet,120g,5% wt) to a reaction vessel, the reaction was carried out sequentially by nitrogen substitution and hydrogen substitution, and the reaction was completed by HPLC monitoring under the conditions of 25.+ -. 10 ℃ and hydrogen pressure of 1.2MPa for 24 hours (Compound IIA < 1%). After nitrogen replacement is carried out on the reaction kettle, the reaction liquid is filtered, the solvent is removed from the filtrate under reduced pressure, acetonitrile (2L, 2V) is added, the solvent is continuously removed by distillation under reduced pressure, then acetonitrile (12L, 12V) is added, and the solution of the compound IIIA is obtained by heating and clearing, and can be directly used for the next reaction, and the yield is close to 100%.
Example 5: synthesis of Compound IVA
Step 1:
to compound IIIA (1.0 Kg,2.0 mol) was added successively cuprous iodide (309 g,1.6 mol), 70% aqueous TBHP (783 g,6.1 mol) at 50.+ -. 5 ℃ and the reaction was stirred for 24 hours and monitored by HPLC to determine the completion of the reaction (compound IIIA < 2%). The temperature of the reaction solution was reduced to 20.+ -. 5 ℃, ethyl acetate (5.0L, 5V) was added, saturated sodium sulfite solution (5.0L, 5V) was added dropwise, stirred, filtered, and saturated sodium chloride solution (3.0L, 3V) was added to the filtrate, stirred, separated, and the organic phase was concentrated under reduced pressure to give an oil, namely 878g of a mixture of the compounds IVA and IVA-a, the percentage ratio of which was shown by HPLC: IIIA/IVA-a=1.6/59.4/32.1. The yield of compound IVA was about 50.7%.
Step 2:
the product from step 1 (20.0 g containing about 6.4g of IVA-a, 12.6 mmol) was taken and methylene chloride (200 mL, 10V) was added, and Dai Sima statin (DMP, 21.4g,50.5mmol, 4.0eq based on IVA-a) was added at 25.+ -. 5 ℃ for 15 hours, and the completion of the reaction was determined by HPLC monitoring (1.6/93.9/0.3% of the compound IVA-a <2% and IIIA/IVA-a). The reaction mixture was washed with 8.4% sodium hydrogencarbonate solution (140 g), separated, the aqueous phase was extracted with methylene chloride (50 mL, 2.5V), the resulting organic phase was washed with 20% sodium sulfite solution (200 g), filtered and separated, the organic phase was washed with 20% sodium chloride solution (150 g), the solvent was removed by concentration of the organic phase, methanol (200 mL, 10V) was added, and the mixture was heated to 60℃and stirred for 0.5 hour, cooled to room temperature and stirred for 2 hours, filtered, the filter cake was washed with methanol (20 mL, 1V), and dried under vacuum at 40℃to give Compound IVA 17.1g, HPLC purity was 97.3%. The total yield of the two steps was 73.1%. The conversion of compound IVA-a to compound IVA was 81.7%.
1 H-NMR(400M,DMSO-d6)δ:7.93-7.95(d,2H),7.62-7.64(t,1H),7.48-7.52(t,3H),5.64(s,1H),4.93(s,1H),3.58(s,3H),2.41-2.49(m,1H),2.34-2.36(m,1H),2.22-2.26(m,1H),2.10-2.13(m,2H),1.86-1.88(m,2H),1.64-1.75(m,6H),1.32-1.55(m,7H),1.20(s,3H),1.24-1.26(m,2H),0.91-0.93(d,3H),0.86(s,3H).m.p.＝124.6-126.4℃.
Example 6: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product from step 1 of example 5 (20.0 g, containing about 6.4g of IVA-a, 12.6 mmol) was taken, methylene chloride (200 mL, 10V) was added, and Dai Sima statin (DMP, 10.7g,25.2mmol,2.0 eq) was added at 25.+ -. 5 ℃ and reacted for 13 hours, after which the reaction was complete (compound IVA-a <2%, percentage ratio IIIA/IVA-a 1.5/93.6/0.2). The reaction solution was washed with 8.4% sodium hydrogencarbonate solution (140 g) under stirring, the aqueous phase was separated, extracted with methylene chloride (50 mL, 2.5V), the resulting organic phase was washed with 20% sodium sulfite solution (200 g), filtered, separated, the organic phase was washed with 20% sodium chloride solution (150 g), the solvent was removed by concentration of the organic phase, methanol (200 mL, 10V) was added, and the mixture was heated to 60℃and stirred for 0.5 hours, cooled to room temperature and stirred for 2 hours, filtered, the solid was rinsed with methanol (20 mL, 1V), dried by suction, and vacuum-dried at 45.+ -. 5℃to give compound IVA 17.2g, with HPLC purity of 97.2%. The total yield of the two steps was 73.5%. The conversion of compound IVA-a to compound IVA was 83.3%.
Example 7: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product of step 1 of example 5 (20.0 g, containing about 6.4g of IVA-a, 12.6 mmol) was taken and acetonitrile (140 mL, 7V), 2-iodoxybenzoic acid (IBX, 5.29g,18.9mmol,1.5 eq) was added and reacted at 70℃for 18 hours and HPLC monitoring determined that the reaction was complete (compound IVA-a <2%, percentage ratio of IIIA/IVA-a 1.2/92.5/1.9). The reaction solution was cooled to room temperature, ethyl acetate (100 mL, 5V) was added, filtration was carried out, the cake was rinsed with ethyl acetate (100 mL, 5V), and 8.4% sodium bicarbonate solution (140 g) was added to the filtrate and stirred for washing, and the solution was separated; the aqueous phase was extracted with ethyl acetate (50 mL, 2.5V), the organic phase was washed with 20% sodium sulfite solution (200 g) with stirring, filtered, the aqueous phase was extracted with ethyl acetate (50 mL, 2.5V), the organic phase was washed with 20% sodium chloride solution (150 g) with stirring, the organic phase was concentrated to remove the solvent, methanol (200 mL, 10V) was added, heated to 65℃and stirred for 1 hour, cooled to 20℃and stirred for 2 hours, filtered, the solid was rinsed with methanol (20 mL, 1V), dried by suction at 45.+ -. 5℃under vacuum to give compound IVA 16.9g with HPLC purity of 98.2%. The total yield of the two steps was 72.2%.
Example 8: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product from step 1 of example 5 (20.0 g, containing about 6.4g of IVA-a, 12.6 mmol) was taken and acetonitrile (140 mL, 7V), 2-iodoxybenzoic acid (IBX, 10.6g,37.8mmol,3.0 eq) was added and reacted at 80℃for 18 hours and HPLC monitoring determined that the reaction was complete (compound IVA-a <2%, percentage ratio IIIA/IVA-a 1.2/92.5/1.9). The reaction mixture was cooled to room temperature, ethyl acetate (100 mL, 5V) was added, and the mixture was filtered, and the cake was rinsed with ethyl acetate (100 mL, 5V); adding 8.4% sodium bicarbonate solution (140 g) into the filtrate, stirring and washing, and separating the solution; the aqueous phase was extracted with ethyl acetate (50 mL, 2.5V), the organic phase was washed with 20% sodium sulfite solution (200 g) with stirring, filtered, the liquid separated and the aqueous phase was extracted with ethyl acetate (50 mL, 2.5V); the organic phase was washed with 20% sodium chloride solution (150 g) with stirring, the organic phase was concentrated to remove the solvent, methanol (200 mL, 10V) was added and heated to 60℃with stirring for 1 hour, cooled to room temperature with stirring for 2 hours, filtered, the solid was rinsed with methanol (20 mL, 1V), pumped down and dried under vacuum at 45.+ -. 5℃to give compound IVA 16.7g with HPLC purity of 98.5%. The total yield of the two steps was 71.4%.
Example 9: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product from step 1 of example 5 (20.0 g, containing about 6.4g,12.6mmol of IVA-a) was taken and reacted with N, N-dimethylformamide (100 mL, 5V), 2-iodoxybenzoic acid (IBX, 7.1g,25.4mmol,2.0 eq.) at room temperature (about 25.+ -. 5 ℃ C.) for 4 hours and monitored by HPLC to determine the completion of the reaction (1.6/94.2/0.1% percent ratio of IVA-a to IVA-a). Ethyl acetate (200 ml,10 v), water (200 ml,10 v) and water are added to the reaction solution, the mixture is stirred and filtered by a filter membrane, the obtained filtrate is separated to obtain an organic phase I, a filter cake is rinsed by ethyl acetate (40 ml,2 v), the rinsed ethyl acetate and the water phase are mixed and stirred, the separated solution is separated to obtain an organic phase II, and the organic phases are combined. Washing with 8.4% sodium bicarbonate solution (130 g) with stirring, and separating; the organic phase was washed with a 20% sodium sulfite solution (140 g) with stirring and separated; the organic phase was washed with 20% sodium chloride solution (140 g), concentrated to remove the solvent, methanol (200 mL, 10V) was added and heated to 55deg.C and stirred for 1 hour, cooled to room temperature and stirred for 3 hours, filtered, the solid was rinsed with methanol (20 mL, 1V), pumped dry and dried under vacuum at 45+ -5deg.C to give compound IVA 17.4g with HPLC purity of 98.4%. The total yield of the two steps was 74.5%.
Example 10: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product from step 1 of example 5 (20.0 g, containing about 6.4g,12.6mmol of IVA-a) was taken and dimethyl sulfoxide (100 mL, 5V), 2-iodoxybenzoic acid (IBX, 5.3g,18.9mmol,1.5 eq) was added and reacted at room temperature for 4 hours and HPLC monitoring determined that the reaction was complete (compound IVA-a <2%, percentage ratio IIIA/IVA-a 1.6/94.3/0.0). Ethyl acetate (200 ml,10 v) was added to the reaction solution, water (200 ml,10 v) was stirred, the obtained filtrate was filtered with a filter membrane, an organic phase I was obtained by separating the filtrate, the filter cake was rinsed with ethyl acetate (40 ml,2 v), the rinsed ethyl acetate was mixed with the water phase, the organic phase II was obtained by separating the filtrate, and the organic phases were combined. Washing with 8.4% sodium bicarbonate solution (130 g) with stirring, and separating; the organic phase was washed with a 20% sodium sulfite solution (140 g) with stirring and separated; the organic phase was washed with 20% sodium chloride solution (140 g) with stirring, then the organic phase was concentrated to remove the solvent, methanol (200 mL, 10V) was added and heated to 60.+ -. 5 ℃ with stirring for 0.5 hours, the temperature was reduced to 20.+ -. 5 ℃ with stirring for 2 hours, the filtration was carried out, the solid was rinsed with methanol (20 mL, 1V), and dried under vacuum at 45.+ -. 5 ℃ to give 17.5g of compound IVA with an HPLC purity of 98.1%. The total yield of the two steps was 74.8%.
Example 11: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product from step 1 of example 5 (20.0 g, containing about 6.4g of IVA-a, 12.6 mmol) was taken, N-dimethylformamide (50 mL, 2.5V) and acetonitrile (50 mL, 2.5V) were added, 2-iodoxybenzoic acid (IBX, 4.23g,15.1mmol,1.2 eq) was added at 60℃and then reacted at 70℃for 12 hours, after which the HPLC monitoring determined that the reaction was complete (compound IVA-a <2%, percentage ratio of IIIA/IVA-a 1.3/94.4/0.1). Ethyl acetate (200 ml,10 v) was added to the reaction solution, water (200 ml,10 v) was stirred, the obtained filtrate was filtered with a filter membrane, an organic phase I was obtained by separating the filtrate, the filter cake was rinsed with ethyl acetate (40 ml,2 v), the rinsed ethyl acetate was mixed with the water phase, the organic phase II was obtained by separating the filtrate, and the organic phases were combined. Washing with 8.4% sodium bicarbonate solution (130 g) with stirring, and separating; the organic phase was washed with a 20% sodium sulfite solution (140 g) with stirring and separated; the organic phase was then washed with 20% sodium chloride solution (140 g) with stirring, the organic phase was concentrated to remove the solvent, methanol (200 mL, 10V) was added and heated to 60.+ -. 5 ℃ with stirring for 0.5 hours, the temperature was reduced to 20.+ -. 5 ℃ with stirring for 2 hours, the filtration was carried out, the solid was rinsed with methanol (20 mL, 1V), and dried under vacuum at 45.+ -. 5 ℃ to give 17.2g of Compound IVA with an HPLC purity of 98.6%. The total yield of the two steps is 73.5%.
Comparative example 1: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product of example 5 (20.0 g, containing about 6.4g of IVA-a, 12.6 mmol) was taken and methylene chloride (200 ml, 10V), 20% aqueous NaClO (12.28 g, containing 33.0mmol of NaClO, 2.6eq based on IVA-a), KBr (0.30 g,2.54mmol, 0.2eq based on IVA-a), TEMPO (0.04 g,0.254mmol,0.02 eq based on IVA-a) was added and the reaction was carried out at 20℃for 20 hours.
Comparative example 2: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product of example 5 (20.0 g containing about 6.4g of IVA-a, 12.6 mmol) was taken and methylene chloride (200 ml, 10V), 20% aqueous NaClO (140.5 g containing 30eq based on IVA-a) KBr (0.30 g,2.54mmol, 0.2eq based on IVA-a), TEMPO (0.04 g,0.254mmol,0.02 times the equivalent based on IVA-a) was added and the reaction was carried out at 30℃for 20 hours, HPLC monitoring showing that the compound IVA-a did not react. Indicating that NaClO is not suitable as an oxidant for this reaction.
Comparative example 3: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product of example 5 (20.0 g containing about 6.4g,12.6mmol of IVA-a) was taken and methylene chloride (200 mL, 10V) was added and iodobenzene diacetic acid (PIDA, 6.1g,18.94mmol, 1.5eq based on IVA-a) was reacted at 30℃for 18 hours, and HPLC monitoring showed that compound IVA-a did not react.
Comparative example 4: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: taking the product of example 5 (20.0 g containing about 6.4g,12.6mmol of IVA-a), adding acetonitrile (200 mL, 10V), controlling the temperature of the system to 50 ℃, adding iodobenzene diacetic acid (PIDA, 6.1g,18.94mmol, 1.5eq based on IVA-a), and reacting at 80 ℃ for 24 hours, wherein HPLC monitoring shows that the content of IVA-a is 20% and the end point of the reaction is not reached; the reaction was prolonged to 62 hours and HPLC monitoring showed the percentage ratio of compound IIIA/IVA-a to be: 1.5/73.5/18.6. IVA-a hardly reacted.
Comparative example 5: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product of example 5 (20.0 g, containing about 6.4g,12.6mmol of IVA-a) was taken and methylene chloride (200 mL, 10V) was added, iodobenzene diacetic acid (PIDA, 8.2g,25.4mmol, 2.0eq based on IVA-a), TEMPO (0.4 g,2.54mmol, 0.2eq based on IVA-a) was reacted at 30℃for 18 hours, and HPLC monitoring showed that compound IVA-a did not react. It is explained that iodobenzene diacetic acid is not suitable as an oxidizing agent for this reaction.
Comparative example 6: synthesis of Compound IVA
Step 1: same as in example 5.
Step 2: the product from step 1 of example 5 (20.0 g, containing about 6.4g,12.6mmol of IVA-a) was taken, methylene chloride (200 mL, 10V) was added, and Dai Sima statin (DMP, 3.5g,8.2mmol,0.65 eq) was added at 25.+ -. 5 ℃ and reacted for 6 hours, the percentage ratio of IIIA/IVA-a in the reaction solution was monitored by HPLC to be 1.6/76.5/18.6. Probably because of insufficient oxidant equivalents, the reactants are not completely converted.
Example 12: synthesis of Compound VA
A solution of compound IVA (500 g,987 mmol), tetrahydrofuran (5.0L, 10V) and 10% Pd/C (dry, 125g,25% wt) were added to the reaction vessel, followed by nitrogen substitution and hydrogen substitution, the system pressure was maintained at 1.2MPa, the reaction system was heated to 55℃and stirred for 24 hours, and HPLC monitoring was carried out to determine that the reaction was complete (compound IVA.ltoreq.3%, percentage ratio of compound IVA/VA/(enol compound VA-a+hydroxy compound VA-b) was 0.015/0.71/0.23.
The reaction solution was filtered, the solvent was removed from the filtrate under reduced pressure, N-dimethylformamide (2.5L, 5V) was added, 2-iodoxybenzoic acid (IBX, 160.3g,572.5mmol, about 2.0eq based on the total amount of VA-a and VA-b) was added, and the reaction was carried out at 25℃for 4 hours, and HPLC monitoring confirmed that the reaction was complete (VA-a and VA-b were both < 1.0%). Ethyl acetate (5.0 l,10 v) was added to the reaction solution, water (5.0 l,10 v) was stirred, the obtained filtrate was filtered with a filter membrane, an organic phase I was obtained by separating the solution, the filter cake was rinsed with ethyl acetate (40 ml,2 v), the rinsed ethyl acetate was mixed with the aqueous phase, and the organic phase II was obtained by separating the solution, and the organic phases were combined. Washing with 8.4% sodium bicarbonate solution (3.25 Kg) with stirring, and separating; the organic phase was washed with 20% sodium sulfite solution (3.5 Kg) under stirring and separated; the organic phase was then washed with 20% sodium chloride solution (3.5 Kg) under stirring, the solvent was removed by concentrating the organic phase, and the organic phase was concentrated to dryness as much as possible. Dissolved in tetrahydrofuran (5.0L, 10V) to obtain a solution.
Adding the solution into a reaction kettle, adding Pd/C recovered before adding, keeping the hydrogen pressure of the system at 1.2MPa and the temperature of 55+/-10 ℃, reacting for 24 hours, and monitoring by HPLC to determine that the reaction is finished (the compound IVA is less than or equal to 1.0%). The reaction body fluid was filtered to remove Pd/C, the filtrate was depressurized to remove the solvent, methylene chloride (1.5L, 3V) and methanol (5L, 10V) were added, methylene chloride was removed under reduced pressure, the system was maintained at 7-8V until methylene chloride was 3% or less, white solid was precipitated during the course, the temperature was lowered to devitrify, the system temperature was maintained at about 0.+ -. 5 ℃ and stirred for 2 hours, filtration was carried out, the filter cake was washed with cold methanol (0.5L, 1V), and air-dried at 50 ℃ to give compound VA427g in 85.1% yield and HPLC purity: 96.2% of VA-a, the main impurity of which is about 3.0%, does not affect the quality of the product.
Compound VA data: 1 H-NMR(400M,DMSO-d6)δ:7.95-7.96(d,2H),7.62-7.66(m,1H),7.49-7.52(t,2H),4.84-4.89(m,1H),3.57(s,3H),2.44-2.54(m,1H),2.30-2.37(m,1H),2.17-2.25(m,1H),1.58-1.92(m,13H),1.22-1.49(m,8H),1.05-1.17(m,2H),1.02(s,3H),0.97(s,3H),0.75-0.77(m,3H).m.p.＝123.2-125.2℃.
example 13: synthesis of Compound VA
A solution of compound IVA (50 g,99 mmol), tetrahydrofuran (500 mL, 10V) and 10% Pd/C (dry, 7.5g,15% wt) were added to the reaction vessel, followed by nitrogen substitution and hydrogen substitution, the system pressure was maintained at 1.0MPa, stirring was carried out at 55.+ -. 10 ℃ for 24 hours, HPLC monitoring was carried out to determine the completion of the reaction (compound IVA.ltoreq.3%, compound VA 70%, enol compound (VA-a) and hydroxy compound (VA-b) 24%, compound IVA 1.5%).
The reaction solution was filtered, the solvent was removed from the filtrate under reduced pressure, N-dimethylformamide (2.5L, 5V) was added, 2-iodoxybenzoic acid (IBX, 25.0g,89.1mmol, about 3.0eq based on total VA-a and VA-b) was added at 25.+ -. 5 ℃ and then reacted at 20-30 ℃ for 4 hours, and HPLC monitoring confirmed the completion of the reaction (VA-a and VA-b were both < 1.0%). Ethyl acetate (500 ml,10 v), water (500 ml,10 v), stirring, filtering with a filter membrane, separating to obtain an organic phase I, washing a filter cake with ethyl acetate (40 ml,2 v), mixing the washed ethyl acetate with an aqueous phase, stirring, separating to obtain an organic phase II, and combining the organic phases. Washing with 8.4% sodium bicarbonate solution (325 g) with stirring, and separating; the organic phase was washed with a 20% sodium sulfite solution (350 g) with stirring and separated; the organic phase was then washed with a 20% sodium chloride solution (350 g) with stirring, the solvent was removed by concentrating the organic phase, and the organic phase was concentrated to dryness as much as possible. Dissolved in tetrahydrofuran (500 mL, 10V) to give a solution.
Adding the solution into a reaction kettle, adding Pd/C recovered before adding, keeping the hydrogen pressure of the system at 1.0MPa and the temperature of 55+/-10 ℃, reacting for 24 hours, and monitoring by HPLC to determine that the reaction is finished (the compound IVA is less than or equal to 1.0%). The reaction was filtered through a filter to remove Pd/C, the solvent was removed from the filtrate under reduced pressure, and methylene chloride (150 mL, 3V) and methanol (500 mL, 10V) were added. Removing dichloromethane under reduced pressure, maintaining the system at 7-8V until dichloromethane is less than or equal to 3%, precipitating white solid, cooling, crystallizing, maintaining the system temperature at-5-0deg.C, stirring for 2 hr, stopping stirring, filtering, washing filter cake with cold methanol (50 mL, 1V), pumping, drying with air at 50deg.C to obtain compound VA42.8g, yield 85.3%, HPLC:96.4% of VA-a, the main impurity, is approximately 2.9%.
Example 14: synthesis of Compound VIA
Compound VA (200 g,393 mmol) was dissolved in tetrahydrofuran (1.2L, 6V) under nitrogen and 1mol/L lithium aluminum tri-tert-butoxide tetrahydrofuran solution (393 mL,393 mmol) was added dropwise at 0.+ -. 5 ℃ and stirring was continued at this temperature for 24 hours. HPLC monitoring confirms that the reaction is completed (compound VA < 3%), dropwise adding 2mol/L hydrochloric acid (600 mL), controlling the temperature within 20 ℃, stirring and separating liquid, extracting and separating organic phase by saturated sodium chloride (200 mL, 1V) and water phase I by tetrahydrofuran (400 mL, 2V) to obtain organic phase II, combining organic phases and adding n-heptane (400 mL, 2V), washing by saturated sodium chloride (400 mL, 2V), separating liquid to obtain organic phase, then decompressing and removing part of solvent, removing water if water liquid is added when about 1400mL (7V) is left, adding n-heptane (2.0L, 10V) for stirring, continuously decompressing and distilling the organic phase to remove tetrahydrofuran (judged by the distilled liquid density being consistent with that of the n-heptane), simultaneously ensuring that the system volume is about 2000mL (10V), stirring for 2 hours at 15+/-10 ℃, filtering, washing filter cake by n-heptane (1V), and drying to obtain the compound A172.6g.
Example 15: synthesis of deoxycholic acid (DCA)
Compound IVA (102.1 g,200 mmol) was dissolved in tetrahydrofuran (715 mL, 7V) with stirring, 4mol/L sodium hydroxide solution (200 mL,800 mmol) was added dropwise at room temperature, and then the reaction was stirred at 60.+ -. 5 ℃ for 36 hours. After the reaction was completed, the pH was adjusted to 9-10 with 7% hydrochloric acid, the organic solvent was removed under reduced pressure, water was added to the system to 2042mL (20V), the system was washed with ethyl acetate (200 mL. Times.2), the pH was adjusted to 1-2 with about 7% hydrochloric acid, stirred for 2 hours at 70.+ -. 10 ℃, cooled to 20.+ -. 10 ℃, filtered, the filter cake was washed with water (510 mL, 5V) 2-3 times, dried to give a solid, the solid was dissolved in methyl t-butyl ether (10200 mL, 10V) at 20.+ -. 5 ℃, stirred for about 12 hours, filtered, the filter cake was washed with methyl t-butyl ether (408 mL, 4V), dried to give deoxycholic acid 72.2g, yield 92.0%.
Example 16: synthesis of Compound IIIB
To the reaction vessel was added compound IIB (50 g,116.7 mmol), tetrahydrofuran (500 mL, 10V) and 10% Pd/C (50% wet,2.5g,5% wt), followed by nitrogen substitution and hydrogen substitution, and then reacted at 20.+ -. 5 ℃ and hydrogen pressure of 1.5MPa for 24 hours, and HPLC monitoring showed that the reaction was completed. After nitrogen gas replacement in the reaction vessel, the reaction solution was filtered, the cake was washed with tetrahydrofuran (50 mL, 1V), and the filtrate was collected and the solvent was removed under reduced pressure to give a white solid compound IIIB
48.2g and 96.0% yield.
Example 17: synthesis of Compound IVB
Step 1:
to the reaction vessel was added compound IIIB (43.1 g,100 mmol) and acetonitrile (430 mL, 10V), and at 45-50℃was added successively cuprous iodide (11.4 g,60 mmol) and 70% aqueous TBHP (51.5 g,400 mmol), followed by stirring for 8 hours and monitoring the completion of the reaction by HPLC (compound IIIB < 2%). The reaction system was cooled to 20.+ -. 5 ℃ and then ethyl acetate (5.0L, 5V), saturated sodium sulfite solution (215 mL, 5V), stirred, filtered, filtrate was added to saturated sodium chloride solution (129 mL, 3V), stirred, separated, and the organic phase was decompressed to remove the solvent to give a mixture of oily substances IVB and IVB-a 36.6g, HPLC showed percentage ratio of compounds: IIIB/IVB-a=1.2/69.5/23.6.
Step 2:
the mixture of step 1 (18.3 g, about 4.3g containing IVB-a, 9.7 mmol) was charged into the reactor, N-dimethylformamide (91.5 mL, 5V) was added, 2-iodoxybenzoic acid (5.4 g, about 19.4mmol, 2.0eq based on IVB-a) was added at 20.+ -. 10 ℃ and then reacted at 20.+ -. 10 ℃ for 4 hours. HPLC monitoring confirmed that the reaction was complete (IVB-a <2%, IIIB/IVB-a% 1.1/95.1/0.0). Ethyl acetate (183 mL, 10V), water (183 mL, 10V) and water were added to the reaction solution, the mixture was stirred, the filtrate was filtered with a filter membrane, an organic phase I was obtained by separating the filtrate, a cake was rinsed with ethyl acetate (36.6 mL, 2V), the rinsed ethyl acetate and the water phase were mixed and stirred, the separated solution was obtained, an organic phase II was obtained, and the organic phases were combined. Washing with 8.4% sodium bicarbonate solution (70 g) with stirring, and separating; the organic phase was washed with 20% sodium sulfite solution (75 g) with stirring and separated; the organic phase was washed with 20% sodium chloride solution (75 g) under stirring, the organic phase was concentrated to remove the solvent, methanol (137.2 mL, 7.5V) was added and heated to 60.+ -. 5 ℃ under stirring to dissolve, pure water (45.7 mL, 2.5V) was added dropwise, the temperature was reduced to 10.+ -. 5 ℃ under stirring for 2 hours, the filtration was carried out, the solid was rinsed with 30% v/v aqueous methanol solution (18.3 mL, 1V), and dried under vacuum at 45.+ -. 5 ℃ to give the compound
17.9g IVB, and the total yield of the two steps is about 80.6%.
Example 18: synthesis of Compound IVB
Step 1: same as in example 17.
Step 2:
the product of step 1 of example 17 (18.3 g, about 4.3g containing IVB-a, 9.7 mmol) was taken and placed in a reaction flask, acetonitrile (183 mL, 10V) was added, 2-iodoxybenzoic acid (8.1 g,29.0mmol, about 3.0 eq) was added at 50.+ -. 10 ℃ and reacted at 75.+ -. 5 ℃ for 24 hours, HPLC monitoring showed that the percentage ratio IIIB/IVB-a was 1.0/94.1/0.9 to reach the end point of the reaction [ (]
IVB-a < 2%). Ethyl acetate (183 mL, 10V) was added, water (183 mL, 10V) was stirred, the filtrate was filtered with a filter membrane, an organic phase I was obtained by separating the filtrate, the cake was rinsed with ethyl acetate (36.6 mL, 2V), the rinsed ethyl acetate was mixed with the water phase, the organic phase II was obtained by separating the filtrate, and the organic phases were combined. Washing with 8.4% sodium bicarbonate solution (70 g) with stirring, and separating; the organic phase was washed with 20% sodium sulfite solution (75 g) with stirring and separated; the organic phase was then washed with 20% sodium chloride solution (75 g) under stirring, the organic phase was concentrated to remove the solvent, methanol (137.2 mL, 7.5V) was added and heated to 60.+ -. 5 ℃ under stirring to dissolve, pure water (45.7 mL, 2.5V) was added dropwise, the temperature was reduced to 10.+ -. 5 ℃ under stirring for 2 hours, the solid was rinsed with 30% v/v aqueous methanol solution (18.3 mL, 1V), and dried under vacuum at 45.+ -. 5 ℃ to give 17.7g of compound IVB, the total yield of the two steps was about 79.7%.
Example 19: synthesis of Compound VB
A solution of compound IVB (25.0 g,56.2 mmol) in tetrahydrofuran (250 mL, 10V) and 10% Pb/C (dry, 5.0g,20% wt) were added to the reaction vessel, followed by nitrogen substitution and hydrogen substitution, and stirring was carried out under a pressure of 1.3MPa at 55.+ -. 10 ℃ for 24 hours, followed by HPLC monitoring to determine the completion of the reaction (compound IVB.ltoreq.3%, compound IVB/compound VB/enol form compound VB-a/hydroxy compound VB-B in a percentage ratio of 0.02/0.69/0.22/0.05).
The reaction mixture was filtered, and after the solvent had been removed from the filtrate under reduced pressure (containing 12.6mmol and 2.8mmol of VB-a and VB-b, respectively, and about 15.4mmol in total), N-dimethylformamide (125 mL, 5V) was added thereto, and 2-iodoxybenzoic acid (12.9 g, about 46.2mmol, 3.0eq based on the sum of VB-a and VB-b) was added at 25.+ -. 5 ℃ for 4 hours, and HPLC monitoring confirmed that the reaction was completed (VB-a and VB-b, respectively < 1.0%). Ethyl acetate (250 mL, 10V) was added, water (250 mL, 10V) was filtered, stirred, filtered with a filter membrane, the resulting filtrate was separated, the cake was rinsed with ethyl acetate (50 mL, 2V), the rinse was mixed with the aqueous phase, extracted, separated, and the organic phases were combined. Washing with 8.4% sodium bicarbonate solution (160 g) with stirring, and separating; the organic phase was washed with a 20% sodium sulfite solution (165 g) with stirring and separated; the organic phase was then washed with 20% sodium chloride solution (165 g) with stirring, the solvent was removed by concentrating the organic phase, and the organic phase was concentrated to dryness as much as possible. Dissolved in tetrahydrofuran (250 mL, 10V) to give a solution.
The solution was added to the reaction vessel, 10%/C (dry, 2.5g,10% wt.) was added and reacted at 55.+ -. 10 ℃ and a hydrogen pressure of 1.3MPa for 24 hours, and the completion of the reaction was monitored by HPLC (compound IVB. Ltoreq.1.0%). The reaction solution was filtered to remove Pd/C, the solvent was removed from the filtrate under reduced pressure, methanol (150 mL, 6V) was added, the solution was heated to dissolve, pure water (75 mL, 3V) was added dropwise, the temperature was lowered to crystallize, stirring was carried out at 15.+ -. 5 ℃ for 2-4 hours, filtration was carried out, the cake was washed with 50% v/v aqueous methanol (25 mL, 1V), suction was carried out, and drying was carried out by air blast at 50℃to give 19.1g of Compound VB in 76.4% yield.
Example 20: synthesis of Compound VIB
Compound VB (20 g,44.8 mmol) was dissolved in tetrahydrofuran (160 mL, 8V) under nitrogen, 1mol/L lithium aluminum tri-tert-butoxide tetrahydrofuran solution (67.2 mL,67.2mmol,1.5 eq) was added dropwise at 0.+ -. 5 ℃ and stirring was continued at that temperature for 24 hours after the addition. HPLC monitoring confirms that the reaction is complete (compound VB < 3%), the reaction solution is cooled, at a temperature not higher than 10 ℃, 2mol/L hydrochloric acid solution (70 mL) is added dropwise, the solution is separated, the organic phase is washed with saturated sodium chloride (20 mL, 1V) and 2mol/L hydrochloric acid solution (20 mL, 1V) in turn, the solution is separated, the organic phase is washed with saturated sodium chloride (40 mL, 2V), then part of the solvent is removed under reduced pressure, n-heptane (200 mL, 10V) is added when about 40mL of volume is left, stirring is carried out, the solution is separated, the tetrahydrofuran is removed by continuous reduced pressure distillation (the density of the distilled liquid is consistent with that of n-heptane), the system volume is about 100mL, stirring is carried out at 10-25 ℃ for 2 hours, filtering is carried out, the filter cake is washed with a small amount of n-heptane, and drying is carried out, thus obtaining 15.8g of compound VI, and the yield is 78.6%.
Example 21: synthesis of deoxycholic acid
Compound VI B (15.0 g,33.5 mmol) was dissolved in tetrahydrofuran (150 mL, 10V) with stirring, 4mol/L sodium hydroxide solution (33.5 mL,134.0 mmol) was added dropwise at room temperature, and the reaction was stirred at 60.+ -. 5 ℃ for 24 hours. After completion of the reaction, tetrahydrofuran was removed under reduced pressure by HPLC, water (120 ml,8 v) was added to the residue, 1mol/L hydrochloric acid was added dropwise to adjust pH to 9-10 at 10-15 ℃, the organic solvent was removed under reduced pressure, the residue was cooled, washed with ethyl acetate (30 ml x 2), adjusted pH to 1-2 with 1mol/L hydrochloric acid, stirred for 2 hours, filtered, the filter cake was washed with water (75 ml,5 v), and air-dried to give deoxycholic acid solid (12.2 g, 84.0%).

Claims (10)

1. A process for preparing compound N comprising: the compound M is oxidized to obtain a compound N,
the reaction formula is as follows:
wherein: p is a hydroxyl protecting group; r is alkyl;
in steroid ringsRepresents a single bond or a double bond; with or without substituents on the steroid ring;
n represents the number of methylene groups, and n is a natural number of 1-5; the methylene group may have a substituent thereon;
the oxidizing agent of the oxidation reaction is selected from the group consisting of iodobenzene diacetic acid, 2-iodoxybenzoic acid, and dessert-martin reagent.
2. The method of claim 1, wherein compound M, N is represented by the following formulas M1, N1, respectively:
Wherein, in the structural formula, P, R,Is as defined in claim 1.
3. The method of claim 1, wherein compound M, N is represented by the following formulas M2, N2, respectively:
wherein, in the structural formula, P,Is as defined in claim 1.
4. A process according to any one of claims 1 to 3, wherein the molar ratio of oxidizing agent to compound M of the oxidation reaction is from (0.5 to 10): 1, preferably from (0.8 to 5): 1.
5. The process according to claim 4, wherein the molar ratio of oxidizing agent to compound M in the oxidation reaction is (1-3): 1.
6. A process as claimed in any one of claims 1 to 3, wherein the reaction solvent of the oxidation reaction comprises a dipolar aprotic solvent.
7. A process as claimed in any one of claims 1 to 3, wherein the reaction solvent for the oxidation reaction is selected from dimethyl sulfoxide, dimethylformamide, N-methylpyrrolidone, dimethylacetamide and acetonitrile.
8. A method for preparing deoxycholic acid intermediate compound IV comprises the steps of oxidizing compound IV-a to obtain compound IV;
the reaction formula is as follows:
wherein P is a hydroxyl protecting group; the oxidizing agent of the oxidation reaction is selected from the group consisting of 2-iodoxybenzoic acid and dess martin reagent.
9. A preparation method of deoxycholic acid intermediate compound V comprises the steps of oxidizing compound V-b to obtain compound V; the reaction formula is as follows:
Wherein P is a hydroxyl protecting group; the oxidizing agent of the oxidation reaction is selected from the group consisting of 2-iodoxybenzoic acid and dess martin reagent.
10. A method for preparing deoxycholic acid, comprising the steps of:
1) The compound II is subjected to hydrogenation reaction to obtain a compound III;
2) Carrying out a first oxidation reaction on the compound III to obtain a mixture containing the compound IV and the compound IV-a; the mixture is subjected to a second oxidation reaction to obtain a compound IV;
3) Carrying out first hydrogenation on the compound IV to obtain a mixture containing compounds V, V-a and V-b; the mixture sequentially undergoes oxidation reaction and second hydrogenation reaction to obtain a compound V;
4) The compound V is subjected to reduction reaction to obtain a compound VI;
5) Deprotection and hydrolysis reaction of the compound VI to obtain deoxycholic acid I or salt thereof;
the reaction formula is as follows:
wherein: p is selected from one of benzyl, P-methoxybenzyl, acetyl, benzoyl, formyl and benzyloxycarbonyl; preferably, P is benzoyl or acetyl;
the method includes one or more of the following features:
in step 1), the hydrogenation is carried out in Pd/C or PtO 2 In the presence of a catalyst;
in step 2), the first oxidation reaction is carried out in the presence of an oxidizing agent and a co-oxidizing agent; the oxidant is tert-butyl hydroperoxide; the co-oxidant is selected from aqueous hypochlorous acid solution, pd/C, pd (OCOCF) 3 ) 2 、Pd(OAc) 2 And CuI;
in step 2), the oxidizing agent used in the second oxidation reaction is selected from the group consisting of 2-iodoxybenzoic acid and dessert-martin oxidizing agent;
in step 3), the first hydrogenation and the second hydrogenation are carried out in Pd/C or PtO 2 In the presence of a catalyst; the oxidizing agent of the oxidation reaction is selected from the group consisting of a dessert-martin oxidizing agent and 2-iodoxybenzoic acid;
in step 4), the reduction reaction is carried out in the presence of lithium aluminum tri-tert-butoxide as a reducing agent;
in step 5), the reaction conditions include an alkali metal hydroxide, an alkali metal alkoxide, or a mixture thereof.
CN202310323916.XA 2023-03-30 2023-03-30 Method for preparing deoxycholic acid intermediate and deoxycholic acid Pending CN116444597A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202310323916.XA CN116444597A (en) 2023-03-30 2023-03-30 Method for preparing deoxycholic acid intermediate and deoxycholic acid

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202310323916.XA CN116444597A (en) 2023-03-30 2023-03-30 Method for preparing deoxycholic acid intermediate and deoxycholic acid

Publications (1)

Publication Number Publication Date
CN116444597A true CN116444597A (en) 2023-07-18

Family

ID=87121231

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202310323916.XA Pending CN116444597A (en) 2023-03-30 2023-03-30 Method for preparing deoxycholic acid intermediate and deoxycholic acid

Country Status (1)

Country Link
CN (1) CN116444597A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115894594A (en) * 2022-11-17 2023-04-04 上海皓元生物医药科技有限公司 Preparation method of deoxycholic acid intermediate

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106146593A (en) * 2015-04-14 2016-11-23 南京诺瑞特医药科技有限公司 A kind of method preparing deoxycholic acid
CN107011401A (en) * 2009-12-18 2017-08-04 凯瑟拉生物制药有限公司 Deoxycholic acid compound and its purification process

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107011401A (en) * 2009-12-18 2017-08-04 凯瑟拉生物制药有限公司 Deoxycholic acid compound and its purification process
CN106146593A (en) * 2015-04-14 2016-11-23 南京诺瑞特医药科技有限公司 A kind of method preparing deoxycholic acid

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115894594A (en) * 2022-11-17 2023-04-04 上海皓元生物医药科技有限公司 Preparation method of deoxycholic acid intermediate
CN115894594B (en) * 2022-11-17 2024-08-30 上海皓元生物医药科技有限公司 Preparation method of deoxycholic acid intermediate

Similar Documents

Publication Publication Date Title
EP3166927B1 (en) Process for the preparation of 4-alkoxy-3-hydroxypicolinic acids
CN116444597A (en) Method for preparing deoxycholic acid intermediate and deoxycholic acid
US10259789B2 (en) Process for the preparation of 4-alkoxy-3-acetoxypicolinic acids
CN109851653B (en) Preparation method of 16 alpha-hydroxy prednisolone
JP2018512066A (en) Method for producing deoxycholic acid
CN102491953A (en) Method for synthesizing florfenicol midbody RT0131
EP3652158B1 (en) Method for preparing pyrimidone compound
CN113874351A (en) Synthetic method of florfenicol
EP0087298B1 (en) Process for producing benzaldehydes
CN103012176A (en) Method for preparing long-chain alkyl 4-carboxyl anionic surfactant
CN107383062A (en) The ANCE of Ceftibuten parent nucleus 7 preparation method
CN109678741B (en) Preparation method of 4-amino-3-fluorobenzoic acid
JP2000502102A (en) Method for preparing 9,11β-epoxide steroid
CN110156696B (en) Preparation method of 1, 4-dichlorophthalazine
EP1456161A1 (en) Method for producing deoxybenzoins
CN110759831B (en) Method for preparing halofuginone intermediate 2-amino-4-bromo-5-chlorobenzoic acid
CN114163309B (en) Preparation method of doravirine intermediate
CN114890902B (en) Preparation method of 2-methyl-3-trifluoromethyl aniline
CN115974809B (en) Method for preparing benzo [ d ] isothiazolin-3 (2H) -ketone through oxygen transfer reaction
JPS636072B2 (en)
CN115215835B (en) Preparation method of 2- (2, 6-dioxopiperidin-3-yl) -1-oxyisoindoline-4-yl) acetic acid
CN114956969B (en) Preparation method of 4-ethoxy-1, 1-trifluoro-3-butene-2-ketone
CN114524802B (en) Synthesis method of quinoline compound
JP3054006B2 (en) Cellobiose derivatives and cellooligomeric derivatives and methods for their production
CN115677590A (en) Preparation method of secnidazole

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
CB02 Change of applicant information
CB02 Change of applicant information

Country or region after: China

Address after: Building F6, 6th Floor, No. 9 Weidi Road, Xianlin Street, Qixia District, Nanjing City, Jiangsu Province, 210018

Applicant after: Nanjing minoway Medical Technology Co.,Ltd.

Address before: Room 637, building a, phase I, Zhongdan Ecological Life Science Industrial Park, no.3-1, xinjinhu Road, Jiangbei new district, Nanjing City, Jiangsu Province, 210018

Applicant before: Nanjing minoway Medical Technology Co.,Ltd.

Country or region before: China