A method for preparing deoxycholic acid intermediate and deoxycholic acid

Abstract

The present invention provides a method for preparing compound N, comprising: compound M is subjected to an oxidation reaction to obtain compound N, and the reaction formula is as follows. Wherein: P is a hydroxyl protecting group; R is an alkyl group; represents a single bond or a double bond in the steroid ring; the steroid ring has or does not have a substituent; n represents the number of methylene groups, and n is a natural number of 1-5; the methylene group has or does not have a substituent; the oxidant of the oxidation reaction is selected from iodophenyl diacetic acid, 2-iodoacylbenzoic acid and Dess-Martin reagent. The method can be used to prepare a key intermediate of deoxycholic acid, does not use a chromium-containing oxidizing agent, is environmentally friendly, has low production cost, good yield, and is suitable for industrial application.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical chemistry, and specifically relates to a method for preparing deoxycholic acid and its intermediates. Background Art

[0002] Deoxycholic acid (Formula I) is a bile acid lacking a hydroxyl group on C-7. It is a free bile acid derived from bile acid losing an oxygen atom. Deoxycholic acid has strong surface activity. After local injection, it can accurately destroy the cell membrane of fat cells, causing the fat cells to rupture and be engulfed by macrophages. However, tissue cells such as skin and muscle will not be lysed because their cell membranes contain a large amount of protein, thereby reducing the dissolution of local subcutaneous fat in a small area. In 2015, the deoxycholic acid injection developed by Kythera was approved by the U.S. Food and Drug Administration as a drug that can be used for injectable lipolysis. The trade name is Kybella, which is used to improve the bulge or fullness of moderate to severe submental fat in adults (double chin).

[0003]

[0004] Currently, deoxycholic acid is mainly derived from animal bodies. Although the cost is low, there is a risk that it may contain animal pathogens and other harmful factors. The prior art has disclosed a method for preparing deoxycholic acid by chemical synthesis.

[0005] CN106083969A discloses a method for the total synthesis of deoxycholic acid, as shown in the following scheme 1. In the method, chromium trioxide is used as an oxidant in the process of synthesizing intermediate 1.10. The oxidant is highly toxic, highly polluting and carcinogenic.

[0006]

[0007] Solution 1

[0008] CN107011401 discloses a method for preparing deoxycholic acid, and the synthetic route is shown in Scheme 2. This method uses an oxidation system of tert-butyl peroxide (TBHP) and sodium hypochlorite in the synthesis of intermediate 2.24, avoiding the use of chromium trioxide. Pyridinium chlorochromate (PCC) is used in the preparation of compound 2.24; by-products are generated in the preparation of 2.33, and the yield is low. In order to improve the reaction yield, it is necessary to further oxidize the by-product 2.34 with pyridinium chlorochromate to obtain 2.33.

[0009]

[0010] Solution 2

[0011] CN106146593 discloses a method for preparing deoxycholic acid, and the synthetic route is shown in Scheme 3. In the process of preparing intermediates 3.4 and 3.5, pyridinium chlorochromate is also needed to be used as an oxidant to improve the conversion of byproducts to target intermediates, so as to improve the yields of these two steps.

[0012]

[0013]

[0014] Solution 3

[0015] Chromium trioxide and pyridinium chlorochromate used in the existing technology are both oxidants containing metallic chromium. Chromium is one of the most toxic pollutants. It is carcinogenic and mutagenic. It has irritating and corrosive effects on the human respiratory tract, can cause rhinitis, tuberculosis, bronchitis and other diseases, and can also induce lung cancer and nasopharyngeal cancer. The discharge of industrial chromium-containing waste gas and wastewater is the main source of man-made chromium pollution. In this regard, my country has successively issued "Several Provisions on the Prevention and Control of Environmental Pollution in the Production and Construction of Chromium Compounds", "Emission Standards for Industrial Pollutants of Chromium and Its Compounds", "Notice on Strengthening the Prevention and Control of Pollution by Chromium-Containing Hazardous Wastes", and included chromium-containing wastes in the "National Hazardous Waste List", with strict index control standards for the discharge of chromium pollutants.

[0016] When enterprises discharge chromium-containing waste gas and wastewater, they need to go through complex and special treatment processes. For example, extraction and washing are usually used to remove chromium-containing components, which consumes a lot of water resources; or chemical precipitation is used to treat the chromium-containing solid waste, which will cause secondary pollution and needs to be entrusted to enterprises with hazardous solid waste disposal qualifications for reprocessing. Not only is the utilization value low, but it is also inconvenient to transport and store, and it also requires high sludge disposal costs. Therefore, regardless of environmental protection and production costs, it is necessary to develop a method that does not use chromium-containing oxidants. Summary of the invention

[0017] The purpose of the present invention is to overcome the above problems and provide a method for oxidizing steroid derivatives, which is particularly suitable for preparing deoxycholic acid intermediates. The method does not require the use of chromium-containing oxidants, is environmentally friendly, has low production costs, high yields, and is suitable for industrial applications.

[0018] The present invention provides a method for preparing compound N, comprising: subjecting compound M to an oxidation reaction to obtain compound N, and the reaction formula is as follows:

[0019]

[0020] in:

[0021] P is a hydroxyl protecting group, which may be selected from benzyl, p-methoxybenzyl, benzoyl, acetyl, formyl and benzyloxycarbonyl; preferably, P is benzoyl or acetyl;

[0022] The steroid ring represents a single bond or a double bond; preferably, in the oxidation reaction The single and double bond types do not change; optionally, the steroid ring has or does not have a substituent;

[0023] R is an alkyl group, preferably a C1-C4 alkyl group, such as methyl, ethyl, propyl, butyl and the like.

[0024] n represents the number of methylene groups, and n is a natural number of 1-5, such as 1, 2, 3, 4, 5; optionally, the methylene group may or may not have a substituent, such as an alkyl group, preferably a C1-C4 alkyl group;

[0025] The oxidant of the oxidation reaction is selected from iodophenyl diacetic acid, 2-iodoacylbenzoic acid (IBX) and Dess-Martin reagent (DMP). Preferably, the oxidant is 2-iodoacylbenzoic acid (IBX) or Dess-Martin reagent (DMP).

[0026] In some embodiments, compounds M and N are represented by the following formulas M1 and N1, respectively:

[0027]

[0028] Among them, P, R, The definition of is as above.

[0029] In other embodiments, compounds M and N are represented by the following formulas M2 and N2, respectively:

[0030]

[0031] Among them, P, R, Same definition as above.

[0032] In other embodiments, compounds M and N are represented by the following formulae M3 and N3, respectively:

[0033]

[0034] Among them, P, Same definition as above.

[0035] In some embodiments, the molar ratio of the oxidant to the compound M in the oxidation reaction may be (0.5-10):1, preferably (0.8-5):1, and more preferably (1-3):1.

[0036] 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.

[0037] In an exemplary embodiment, a method for preparing a deoxycholic acid intermediate compound IV is provided, comprising subjecting compound IV-a to an oxidation reaction to obtain compound IV;

[0038] The reaction formula is as follows:

[0039]

[0040] Wherein, P is a hydroxyl protecting group, for example, P is selected from benzyl, p-methoxybenzyl, benzoyl, acetyl, formyl and benzyloxycarbonyl; preferably, P is benzoyl or acetyl;

[0041] The oxidant of the oxidation reaction is selected from 2-iodoxybenzoic acid and Dess-Martin reagent.

[0042] In some embodiments, the molar ratio of compound IV-a to the oxidant may be (0.5-10):1, preferably (0.8-5):1, and more preferably (1-3):1.

[0043] 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.

[0044] In another exemplary embodiment, a method for preparing a deoxycholic acid intermediate compound V is provided, comprising subjecting compound Vb to an oxidation reaction to obtain compound V;

[0045] The reaction formula is as follows:

[0046]

[0047] P is a hydroxyl protecting group, for example, P is selected from one of benzyl, p-methoxybenzyl, benzoyl, acetyl, formyl, and benzyloxycarbonyl; more preferably, P is benzoyl or acetyl;

[0048] The oxidant of the oxidation reaction is selected from 2-iodoxybenzoic acid and Dess-Martin reagent.

[0049] In some embodiments, the molar ratio of the oxidant to the compound Vb may be (0.5-10):1, preferably (0.8-5):1, and more preferably (1-3):1.

[0050] 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.

[0051] The present invention further provides a method for preparing deoxycholic acid, comprising the following steps:

[0052] 1) Compound II is subjected to hydrogenation reaction to obtain compound III;

[0053] 2) Compound III is subjected to a first oxidation reaction to obtain a mixture containing Compound IV and Compound IV-a; the mixture is subjected to a second oxidation reaction to obtain Compound IV;

[0054] 3) Compound IV is subjected to a first hydrogenation reaction to obtain a mixture containing compounds V, V-a and V-b; the mixture is subjected to an oxidation reaction and a second hydrogenation reaction in sequence to obtain compound V;

[0055] 4) Compound V is subjected to reduction reaction to obtain compound VI;

[0056] 5) Compound VI is subjected to deprotection and hydrolysis to obtain deoxycholic acid I or its salt;

[0057] The reaction formula is as follows:

[0058]

[0059] Wherein, P is a hydroxyl protecting group; it can be selected from benzyl, p-methoxybenzyl, acetyl, benzoyl, formyl and benzyloxycarbonyl; preferably, P is benzoyl or acetyl.

[0060] In some embodiments, in step 1), the hydrogenation reaction is carried out in the presence of a Pd / C or PtO2 catalyst; preferably a Pd / C catalyst.

[0061] In some embodiments, in step 2), the first oxidation reaction is carried out in the presence of an oxidant and a co-oxidant. The oxidant is preferably tert-butyl hydroperoxide (TBHP). The co-oxidant is selected from aqueous hypochlorous acid solution, Pd / C, Pd(OCOCF3)2, Pd(OAc)2 and CuI; preferably aqueous hypochlorous acid solution or CuI; more preferably CuI.

[0062] In some embodiments, in step 2), the oxidant used in the second oxidation reaction is selected from 2-iodoacylbenzoic acid and Dess-Martin reagent.

[0063] In some embodiments, in step 2), the molar ratio of the oxidant used in the second oxidation reaction to compound IV-a can be (0.8-5):1, for example (1-3):1, such as 1:1, 1.5:1, 2:1, etc.

[0064] In some embodiments, in step 2), the reaction solvent of the second oxidation reaction includes a dipolar aprotic solvent, such as dimethyl sulfoxide (DMSO), dimethylformamide (DMF), N-methylpyrrolidone (NMP), dimethylacetamide (DMA), acetonitrile, etc.

[0065] In some embodiments, in step 3), compound IV is subjected to a first hydrogenation reaction to obtain a mixture containing compounds V, V-a and V-b; the mixture containing compounds V, V-a and V-b is subjected to an oxidation reaction to obtain a mixture essentially containing compounds IV and V; the mixture essentially containing compounds IV and V is subjected to a second hydrogenation reaction to obtain compound V.

[0066] In some embodiments, in step 3), the first hydrogenation reaction and the second hydrogenation reaction are carried out in the presence of a Pd / C or PtO2 catalyst; preferably a Pd / C catalyst.

[0067] In some embodiments, in step 3), the oxidizing agent used in the oxidation reaction is selected from 2-iodoacylbenzoic acid and Dess-Martin reagent.

[0068] In some embodiments, in step 3), 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, etc.

[0069] In some embodiments, in step 3), the oxidant undergoes oxidation reaction with both compounds V-a and V-b. The molar ratio of the oxidant to the sum of compounds V-a and V-b is (0.5-10):1, preferably (0.8-5):1, for example (1-3):1.

[0070] In some embodiments, in step 4), the reducing agent used in the reduction reaction is lithium tri-tert-butoxyaluminum hydride (LiAl(OtBu)3).

[0071] In some embodiments, in step 5), the deprotection and hydrolysis reactions can be carried out according to known methods. For example, the deprotection and hydrolysis conditions include 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 acid treatment to obtain deoxycholic acid. In certain embodiments, the acid treatment is omitted to obtain the corresponding salt. The alkaline earth metal hydroxide can be lithium hydroxide (LiOH) or sodium hydroxide (NaOH).

[0072] The method of the present invention attempts to use a new oxidant (such as IBX and DMP) to efficiently complete the oxidation reaction, avoiding the harm to the environment and humans caused by the use of chromium-containing oxidants, and does not require a large amount of cost to treat chromium-containing waste liquid, which significantly reduces the production cost. The method can be used to prepare intermediate compounds IV and V of deoxycholic acid, is environmentally friendly, convenient for post-treatment, low in cost, high in yield, and suitable for industrial production. DETAILED DESCRIPTION

[0073] The present invention is described in detail below by way of examples. It should be understood that the methods in the examples are for illustration purposes only and do not constitute any limitation of the present invention. Unless otherwise specified, the materials used in the examples are commercially available. Unless otherwise specified, the experimental methods used are all conventional methods. In the examples, the completion of the reaction is monitored by HPLC, and the mass percentage of the compound is obtained by area normalization method. Unless otherwise specified, the percentages of the solutions used in the examples are all the mass percentages of the contained solutes.

[0074] The steroid structure skeleton numbering in the present invention follows the general rules:

[0075]

[0076] Unless otherwise indicated, the backbone only indicates the position of carbon atoms. If not specifically indicated, the substituent groups on the backbone structure of the support may include R configuration, S configuration, or a mixture thereof.

[0077] The "Pd / C" catalyst in the present invention refers to a type of catalyst in which Pd is loaded on C, which can be used for catalytic hydrogenation reactions. The "Pd / C" catalyst may be water-free or contain a certain amount of water. Various specifications of Pd / C catalysts (e.g., 5% Pd, 10% Pd) can be used in the present invention. The amount of the catalyst can be added according to conventional methods.

[0078] In the present invention, "hydroxy protecting group P" refers to a protecting group suitable for preventing hydroxyl side reactions well known in the field of organic synthesis. For example, many protecting groups are described in TW Greene and GM Wuts, Protecting Groups in Organic Synthesis, 3rd edition, Wiley, New York, 1999 and references cited therein. Representative hydroxy protecting groups include, but are not limited to: alkyl (such as methyl, ethyl and tert-butyl); acyl (such as formyl, acetyl, benzoyl); arylmethyl (such as benzyl Bn, p-methoxybenzyl PMB, 9-fluorenylmethyl Fm, diphenylmethyl DPM; silyl (such as trimethylsilyl TMS, tert-butyldimethylsilyl TBS), etc. Preferably, the hydroxy protecting group is acetyl or benzoyl.

[0079] The "alkyl" of the present invention refers to a saturated monovalent hydrocarbon group, preferably a C1-C4 alkyl group, such as methyl, ethyl, propyl, isopropyl, butyl and the like.

[0080] The intermediate compound IV of deoxycholic acid in the present invention can be prepared by using compound II or compound III as the starting material. The preparation methods of compound II and compound III have been disclosed, for example, see patent applications CN106146593A, CN101711254A, CN107011401A, CN102512035A, CN103906517A or CN114805463A. As an example, compound IIA in the following examples is prepared by referring to the method of CN106146593A. Compound IIB is prepared by referring to the method of CN101711254A.

[0081] Example 1: Synthesis of Compound IIIA

[0082]

[0083] Compound ⅡA (50 g, 0.1 mol), tetrahydrofuran (500 mL, 10 V), and 10% Pd / C (50% wet, 2.5 g, 5% wt) were added to the reactor, and then nitrogen and hydrogen replacement were performed in sequence. The reaction was carried out at a temperature of 25±10°C and a hydrogen pressure of 1.2 MPa for 24 hours. The reaction was completed by HPLC monitoring (Compound ⅡA <1%). After nitrogen replacement of the reactor, the reaction liquid was filtered, the filtrate was decompressed to remove the solvent, acetonitrile (150 mL, 3 V) was added, heated to dissolve, cooled to -5±5°C, stirred for 6 hours, filtered, and washed with acetonitrile (25 mL, 0.5 V) precooled to -15±5°C to obtain 47.2 g of compound ⅢA, with a yield of 94.0% and a HPLC purity of 99.5%.

[0084] 1 HNMR (400MHz, CDCl3) 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,C9),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,C3),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 ). Mass spectrometry (ES(+)) measured the product [M+NH4] + The mass-to-charge ratio of the peak is 510.6.

[0085] Example 2: Synthesis of Compound IIIA

[0086] After adding compound ⅡA (50g, 0.1mol), tetrahydrofuran (500mL, 10V), and 10% Pd / C (50% wet., 2.5g, 5% wt) to the reactor, the device was sealed, nitrogen replacement and hydrogen replacement were carried out successively, and the reaction was carried out at a temperature of 30±10℃ and a hydrogen pressure of 1.3MPa for 16 hours. The reaction was completed by HPLC monitoring (compound ⅡA<1%). After nitrogen replacement of the reactor, the reaction liquid was filtered, the filtrate was decompressed to remove the solvent, and acetonitrile (150mL, 3V) was added to heat and dissolve, cooled to -5±5℃, stirred for 5 hours, filtered, and washed with acetonitrile (25mL, 0.5V) precooled to -15±5℃ to obtain 47.8g of compound ⅢA, with a yield of 95.2% and a HPLC purity of 99.3%.

[0087] Example 3 Synthesis of Compound IIIA

[0088] Compound IIA (1.0 Kg, 2.0 mol), tetrahydrofuran (5 L, 5 V) and 10% Pd / C (50% wet, 120 g, 12% wt) were added to the reactor, and nitrogen and hydrogen replacement were performed in sequence, and then the reaction was carried out at 25±10°C and a hydrogen pressure of 1.0 MPa for 24 hours. The reaction was completed as determined by HPLC monitoring (Compound IIA <1%). After nitrogen replacement of the reactor, the reaction liquid was filtered, the filtrate was decompressed to remove the solvent, acetonitrile (2 L, 2 V) was added, and the solvent was continued to be distilled under reduced pressure, followed by the addition of acetonitrile (12 L, 12 V) to heat and dissolve, cool to -5±5°C, stir for 5 hours, filter, and wash with acetonitrile (500 mL, 0.5 V) precooled to -15±5°C to obtain 963.6 g of compound IIIA, with a yield of 96.0% and a HPLC purity of 99.4%.

[0089] Example 4: Synthesis of Compound IIIA

[0090] After adding compound ⅡA (1.0Kg, 2.0mol), tetrahydrofuran (10L, 10V) and 10% Pd / C (50% wet, 120g, 5%wt) to the reactor, nitrogen replacement and hydrogen replacement were carried out in sequence, and the reaction was carried out at 25±10°C and a hydrogen pressure of 1.2MPa for 24 hours. The reaction was completed by HPLC monitoring (compound ⅡA<1%). After nitrogen replacement of the reactor, the reaction liquid was filtered, the filtrate was decompressed to remove the solvent, acetonitrile (2L, 2V) was added, and the solvent was continued to be distilled under reduced pressure, and then acetonitrile (12L, 12V) was added and heated to dissolve, and a solution of compound ⅢA was obtained, which can be directly used in the next step reaction, and the yield is close to 100%.

[0091] Example 5: Synthesis of Compound IVA

[0092]

[0093] Step 1:

[0094] At 50±5°C, add cuprous iodide (309g, 1.6mol) and 70% TBHP aqueous solution (783g, 6.1mol) to compound IIIA (1.0Kg, 2.0mol) in sequence, stir and react for 24 hours, and monitor the reaction with HPLC to determine that the reaction is complete (compound IIIA<2%). Lower the temperature of the reaction solution to 20±5°C, add ethyl acetate (5.0L, 5V), dropwise add saturated sodium sulfite solution (5.0L, 5V), stir, filter, add saturated sodium chloride solution (3.0L, 3V) to the filtrate, stir, separate, and concentrate the organic phase under reduced pressure to obtain an oily substance, i.e., a mixture of compound IVA and IVA-a (878g). HPLC shows that the percentage ratio of the compounds is: IIIA / IVA / IVA-a=1.6 / 59.4 / 32.1. The yield of compound IVA is about 50.7%.

[0095] Step 2:

[0096] The product obtained in step 1 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol) was added with dichloromethane (200 mL, 10 V), and desmartine reagent (DMP, 21.4 g, 50.5 mmol, based on 4.0 eq of IVA-a) was added at 25±5° C. and reacted for 15 hours. The reaction was completed by HPLC monitoring (compound IVA-a<2%, and the percentage ratio of IIIA / IVA / IVA-a was 1.6 / 93.9 / 0.3). 8.4% sodium bicarbonate solution (140 g) was added to the reaction solution for washing, and the solution was separated. The aqueous phase was extracted with dichloromethane (50 mL, 2.5 V), and the obtained organic phase was washed with 20% sodium sulfite solution (200 g). After filtration, the organic phase was separated, and the organic phase was washed with 20% sodium chloride solution (150 g). The organic phase was concentrated to remove the solvent, and methanol (200 mL, 10 V) was added and heated to 60° C. and stirred for 0.5 hours, cooled to room temperature and stirred for 2 hours, filtered, and the filter cake was washed with methanol (20 mL, 1 V) and vacuum dried at 40° C. to obtain 17.1 g of compound IVA with an HPLC purity of 97.3%. The total yield of the two steps was 73.1%. The conversion rate of compound IVA-a to compound IVA was 81.7%.

[0097] 1H-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),2.22-2.2 6(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).mp=124.6-126.4℃.

[0098]

[0099] Example 6: Synthesis of Compound IVA

[0100] Step 1: Same as Example 5.

[0101] Step 2: Take the product obtained in Step 1 of Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add dichloromethane (200 mL, 10 V), add desmartine reagent (DMP, 10.7 g, 25.2 mmol, 2.0 eq) at 25±5°C, react for 13 hours, and monitor the completion of the reaction by HPLC (compound IVA-a <2%, the percentage ratio of IIIA / IVA / IVA-a is 1.5 / 93.6 / 0.2). The reaction solution was added with 8.4% sodium bicarbonate solution (140 g) and stirred and washed. The aqueous phase was extracted with dichloromethane (50 mL, 2.5 V), and the obtained organic phase was washed with 20% sodium sulfite solution (200 g). After filtration, the organic phase was separated and washed with 20% sodium chloride solution (150 g). The organic phase was concentrated to remove the solvent, methanol (200 mL, 10 V) was added, heated to 60 ° C and stirred for 0.5 hours, cooled to room temperature and stirred for 2 hours, filtered, and the solid was rinsed with methanol (20 mL, 1 V), dried, and vacuum dried at 45 ± 5 ° C to obtain 17.2 g of compound IVA with an HPLC purity of 97.2%. The total yield of the two steps was 73.5%. The conversion rate of compound IVA-a to compound IVA was 83.3%.

[0102] Example 7: Synthesis of Compound IVA

[0103] Step 1: Same as Example 5.

[0104] Step 2: Take the product of Step 1 in Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add acetonitrile (140 mL, 7V), 2-iodoacylbenzoic acid (IBX, 5.29 g, 18.9 mmol, 1.5 eq), react at 70°C for 18 hours, and the reaction is determined to be complete by HPLC monitoring (compound IVA-a <2%, the percentage ratio of IIIA / IVA / IVA-a is 1.2 / 92.5 / 1.9). The reaction solution was cooled to room temperature, ethyl acetate (100 mL, 5V) was added, filtered, the filter cake was rinsed with ethyl acetate (100 mL, 5V), 8.4% sodium bicarbonate solution (140 g) was added to the filtrate, stirred and washed, and the liquid was separated; the aqueous phase was extracted with ethyl acetate (50 mL, 2.5V), the organic phase was stirred and washed with 20% sodium sulfite solution (200 g), filtered, separated, the aqueous phase was extracted with ethyl acetate (50 mL, 2.5V), the organic phase was stirred and washed with 20% sodium chloride solution (150 g), the organic phase was concentrated to remove the solvent, methanol (200 mL, 10V) was added, heated to 65°C and stirred for 1 hour, cooled to 20°C and stirred for 2 hours, filtered, the solid was rinsed with methanol (20 mL, 1V), dried, and vacuum dried at 45±5°C to obtain 16.9 g of compound IVA with an HPLC purity of 98.2%. The total yield of the two steps was 72.2%.

[0105] Example 8: Synthesis of Compound IVA

[0106] Step 1: Same as Example 5.

[0107] Step 2: Take the product obtained in Step 1 of Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add acetonitrile (140 mL, 7V), 2-iodoacylbenzoic acid (IBX, 10.6 g, 37.8 mmol, 3.0 eq), react at 80°C for 18 hours, and monitor the completion of the reaction by HPLC (compound IVA-a <2%, percentage ratio of IIIA / IVA / IVA-a is 1.2 / 92.5 / 1.9). The reaction solution was cooled to room temperature, ethyl acetate (100 mL, 5V) was added, filtered, and the filter cake was rinsed with ethyl acetate (100 mL, 5V); 8.4% sodium bicarbonate solution (140 g) was added to the filtrate, stirred and washed, and the liquids were separated; the aqueous phase was extracted with ethyl acetate (50 mL, 2.5V), the organic phase was stirred and washed with 20% sodium sulfite solution (200 g), filtered, separated, and the aqueous phase was extracted with ethyl acetate (50 mL, 2.5V); the organic phase was stirred and washed with 20% sodium chloride solution (150 g), the organic phase was concentrated to remove the solvent, methanol (200 mL, 10V) was added, heated to 60°C and stirred for 1 hour, cooled to room temperature and stirred for 2 hours, filtered, the solid was rinsed with methanol (20 mL, 1V), dried, and vacuum dried at 45±5°C to obtain 16.7 g of compound IVA with an HPLC purity of 98.5%. The total yield of the two steps was 71.4%.

[0108] Example 9: Synthesis of Compound IVA

[0109] Step 1: Same as Example 5.

[0110] Step 2: Take the product obtained in step 1 of Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add N,N-dimethylformamide (100 mL, 5 V), 2-iodoacylbenzoic acid (IBX, 7.1 g, 25.4 mmol, 2.0 eq), react at room temperature (about 25±5°C) for 4 hours, and HPLC monitoring determines that the reaction is complete (compound IVA-a <2%, IIIA / IVA / IVA-a percentage ratio is 1.6 / 94.2 / 0.1). Add ethyl acetate (200 mL, 10 V) and water (200 mL, 10 V) to the reaction solution, stir, filter with a filter membrane, and separate the filtrate to obtain an organic phase I. Rinse the filter cake with ethyl acetate (40 mL, 2 V), mix the rinsed ethyl acetate with the water phase, stir, and separate to obtain an organic phase II, and combine the organic phases. 8.4% sodium bicarbonate solution (130 g) was used for stirring and washing, and the liquid was separated; the organic phase was stirred and washed with 20% sodium sulfite solution (140 g), and the liquid was separated; the organic phase was washed with 20% sodium chloride solution (140 g), concentrated to remove the solvent, methanol (200 mL, 10 V) was added, heated to 55° C. and stirred for 1 hour, cooled to room temperature and stirred for 3 hours, filtered, and the solid was rinsed with methanol (20 mL, 1 V), dried, and vacuum dried at 45±5° C. to obtain 17.4 g of compound IVA, with an HPLC purity of 98.4%. The total yield of the two steps was 74.5%.

[0111] Example 10: Synthesis of Compound IVA

[0112] Step 1: Same as Example 5.

[0113] Step 2: Take the product obtained in step 1 of Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add dimethyl sulfoxide (100 mL, 5 V), 2-iodoacylbenzoic acid (IBX, 5.3 g, 18.9 mmol, 1.5 eq), react at room temperature for 4 hours, and HPLC monitoring determines that the reaction is complete (compound IVA-a <2%, IIIA / IVA / IVA-a percentage ratio is 1.6 / 94.3 / 0.0). Add ethyl acetate (200 mL, 10 V) and water (200 mL, 10 V) to the reaction solution, stir, filter with a filter membrane, and separate the filtrate to obtain an organic phase I. Rinse the filter cake with ethyl acetate (40 mL, 2 V), mix the rinsed ethyl acetate with the aqueous phase, stir, and separate to obtain an organic phase II, and combine the organic phases. 8.4% sodium bicarbonate solution (130 g) was used for stirring and washing, and the liquid was separated; the organic phase was stirred and washed with 20% sodium sulfite solution (140 g), and the liquid was separated; the organic phase was stirred and washed with 20% sodium chloride solution (140 g), and then the organic phase was concentrated to remove the solvent, methanol (200 mL, 10 V) was added, heated to 60±5°C and stirred for 0.5 hours, cooled to 20±5°C and stirred for 2 hours, filtered, and the solid was rinsed with methanol (20 mL, 1 V), dried, and vacuum dried at 45±5°C to obtain 17.5 g of compound IVA, with an HPLC purity of 98.1%. The total yield of the two steps was 74.8%.

[0114] Example 11: Synthesis of Compound IVA

[0115] Step 1: Same as Example 5.

[0116] Step 2: Take the product obtained in step 1 of Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add N,N-dimethylformamide (50 mL, 2.5 V) and acetonitrile (50 mL, 2.5 V), add 2-iodoacylbenzoic acid (IBX, 4.23 g, 15.1 mmol, 1.2 eq) at 60°C, then react at 70°C for 12 hours, and HPLC monitoring determines that the reaction is complete (compound IVA-a <2%, IIIA / IVA / IVA-a percentage ratio is 1.3 / 94.4 / 0.1). Add ethyl acetate (200 mL, 10 V) and water (200 mL, 10 V) to the reaction solution, stir, filter with a filter membrane, and separate the filtrate to obtain an organic phase I. Rinse the filter cake with ethyl acetate (40 mL, 2 V), mix the rinsed ethyl acetate with the water phase, stir, and separate to obtain an organic phase II, and combine the organic phases. 8.4% sodium bicarbonate solution (130 g) was used for stirring and washing, and the liquid was separated; the organic phase was stirred and washed with 20% sodium sulfite solution (140 g), and the liquid was separated; the organic phase was stirred and washed with 20% sodium chloride solution (140 g), the organic phase was concentrated to remove the solvent, methanol (200 mL, 10 V) was added, heated to 60±5°C and stirred for 0.5 hours, cooled to 20±5°C and stirred for 2 hours, filtered, the solid was rinsed with methanol (20 mL, 1 V), dried, and vacuum dried at 45±5°C to obtain 17.2 g of compound IVA with an HPLC purity of 98.6%. The total yield of the two steps was 73.5%.

[0117] Comparative Example 1: Synthesis of Compound IVA

[0118] Step 1: Same as Example 5.

[0119] Step 2: Take the product in Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add dichloromethane (200 ml, 10 V), 20% NaClO aqueous solution (12.28 g, containing 33.0 mmol of NaClO, based on 2.6 eq of IVA-a), KBr (0.30 g, 2.54 mmol, based on 0.2 eq of IVA-a), TEMPO (0.04 g, 0.254 mmol, based on 0.02 eq of IVA-a), and react at 20° C. for 20 hours. HPLC monitoring shows that compound IVA-a does not react.

[0120] Comparative Example 2: Synthesis of Compound IVA

[0121] Step 1: Same as Example 5.

[0122] Step 2: Take the product in Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add dichloromethane (200 ml, 10 V), 20% NaClO aqueous solution (140.5 g, containing 377 mmol of NaClO, based on 30 eq of IVA-a), KBr (0.30 g, 2.54 mmol, based on 0.2 eq of IVA-a), TEMPO (0.04 g, 0.254 mmol, 0.02 equivalents, based on the equivalent of IVA-a), react at 30° C. for 20 hours, HPLC monitoring shows that compound IVA-a does not react. This indicates that NaClO is not suitable as an oxidant for this reaction.

[0123] Comparative Example 3: Synthesis of Compound IVA

[0124] Step 1: Same as Example 5.

[0125] Step 2: Take the product in Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add dichloromethane (200 mL, 10 V), iodophenyldiacetic acid (PIDA, 6.1 g, 18.94 mmol, based on 1.5 eq of IVA-a), and react at 30° C. for 18 hours. HPLC monitoring shows that compound IVA-a does not react.

[0126] Comparative Example 4: Synthesis of Compound IVA

[0127] Step 1: Same as Example 5.

[0128] Step 2: Take the product in Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add acetonitrile (200 mL, 10 V), control the system temperature to 50° C., add iodophenyl diacetic acid (PIDA, 6.1 g, 18.94 mmol, based on 1.5 eq of IVA-a), react at 80° C. for 24 hours, HPLC monitoring shows that the content of IVA-a is 20%, and the reaction end point has not been reached; extend the reaction to 62 hours, HPLC monitoring shows that the percentage ratio of compound IIIA / IVA / IVA-a is: 1.5 / 73.5 / 18.6. IVA-a hardly reacts.

[0129] Comparative Example 5: Synthesis of Compound IVA

[0130] Step 1: Same as Example 5.

[0131] Step 2: Take the product in Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add dichloromethane (200 mL, 10 V), iodophenyldiacetic acid (PIDA, 8.2 g, 25.4 mmol, based on 2.0 eq of IVA-a), TEMPO (0.4 g, 2.54 mmol, based on 0.2 eq of IVA-a), and react at 30° C. for 18 hours. HPLC monitoring shows that compound IVA-a does not react. This indicates that iodophenyldiacetic acid is not suitable as an oxidant for this reaction.

[0132] Comparative Example 6: Synthesis of Compound IVA

[0133] Step 1: Same as Example 5.

[0134] Step 2: Take the product obtained in step 1 of Example 5 (20.0 g, containing about 6.4 g of IVA-a, 12.6 mmol), add dichloromethane (200 mL, 10 V), add Desmartine reagent (DMP, 3.5 g, 8.2 mmol, 0.65 eq) at 25±5°C, react for 6 hours, and the percentage ratio of compound IIIA / IVA / IVA-a in the reaction solution monitored by HPLC is 1.6 / 76.5 / 18.6. It may be due to insufficient oxidant equivalents that the reactants were not completely converted.

[0135] Example 12: Synthesis of Compound VA

[0136]

[0137] Compound IVA (500 g, 987 mmol), a solution of tetrahydrofuran (5.0 L, 10 V), and 10% Pd / C (dry, 125 g, 25% wt) were added to a reaction kettle, and nitrogen and hydrogen replacement were performed successively. The system pressure was maintained at 1.2 MPa, and the reaction system was heated to 55° C. and stirred for 24 hours. The completion of the reaction was determined by HPLC monitoring (compound IVA ≤ 3%, and the percentage ratio of compound IVA / VA / (enol compound VA-a + hydroxy compound VA-b) was 0.015 / 0.71 / 0.23).

[0138] The reaction solution was filtered, the filtrate was decompressed to remove the solvent, N,N-dimethylformamide (2.5L, 5V) was added, 2-iodoacylbenzoic 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°C for 4 hours. HPLC monitoring confirmed that the reaction was complete (VA-a and VA-b were both <1.0%). Ethyl acetate (5.0L, 10V) and water (5.0L, 10V) were added to the reaction solution, stirred, filtered with a filter membrane, and the obtained filtrate was separated to obtain an organic phase I. The filter cake was rinsed with ethyl acetate (40mL, 2V), and the rinsed ethyl acetate was mixed with the aqueous phase, stirred, and separated to obtain an organic phase II, and the organic phases were combined. Use 8.4% sodium bicarbonate solution (3.25Kg) to stir and wash, and separate the liquids; stir and wash the organic phase with 20% sodium sulfite solution (3.5Kg), and separate the liquids; stir and wash the organic phase with 20% sodium chloride solution (3.5Kg), and concentrate the organic phase to remove the solvent and concentrate it to dryness as much as possible. Dissolve it with tetrahydrofuran (5.0L, 10V) to obtain a solution.

[0139] The solution was added to the reactor, and the previously recovered Pd / C was added. The system hydrogen pressure was maintained at 1.2MPa and the temperature was 55±10°C. The reaction was carried out for 24 hours. The reaction was completed by HPLC monitoring (Compound IVA≤1.0%). The reaction liquid was filtered to remove Pd / C, the filtrate was decompressed to remove the solvent, dichloromethane (1.5L, 3V) and methanol (5L, 10V) were added, and dichloromethane was removed under reduced pressure. The system was maintained at 7-8V until dichloromethane was less than or equal to 3%. A white solid was precipitated during the process. The temperature was lowered for crystallization. The system temperature was maintained at about 0±5°C and stirred for 2 hours. The filter cake was washed with cold methanol (0.5L, 1V) and dried with air at 50°C to obtain 427g of compound VA, with a yield of 85.1%, HPLC purity: 96.2%, and the main impurity was VA-a containing about 3.0%, which did not affect the quality of the product.

[0140] Compound VA data: 1 H-NMR(400M,DMSO-d6)δ:7.95-7.96(d,2H),7.62-7.66(m,1H),7.49-7.5 2(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),1.05-1.1 7(m,2H),1.02(s,3H),0.97(s,3H),0.75-0.77(m,3H).mp=123.2-125.2℃.

[0141] Example 13: Synthesis of Compound VA

[0142] Compound IVA (50 g, 99 mmol), a solution of tetrahydrofuran (500 mL, 10 V), and 10% Pd / C (dry, 7.5 g, 15% wt) were added to a reaction kettle, and nitrogen and hydrogen replacement were carried out successively. The system pressure was maintained at 1.0 MPa, and the mixture was stirred at 55±10°C for 24 hours. The completion of the reaction was determined by HPLC monitoring (Compound IVA ≤ 3%. Compound VA accounted for 70%, enol compound (VA-a) and hydroxy compound (VA-b) accounted for 24%, and Compound IVA accounted for 1.5%).

[0143] The reaction solution was filtered, the filtrate was decompressed to remove the solvent, N,N-dimethylformamide (2.5L, 5V) was added, 2-iodoacylbenzoic acid (IBX, 25.0g, 89.1mmol, about 3.0eq based on the total amount of VA-a and VA-b) was added at 25±5°C, and then reacted at 20-30°C for 4 hours. HPLC monitoring confirmed that the reaction was complete (VA-a and VA-b were both <1.0%). Ethyl acetate (500mL, 10V) and water (500mL, 10V) were added to the reaction solution, stirred, and filtered with a filter membrane. The obtained filtrate was separated to obtain an organic phase I. The filter cake was rinsed with ethyl acetate (40mL, 2V), and the rinsed ethyl acetate was mixed with the water phase, stirred, and separated to obtain an organic phase II, and the organic phases were combined. Use 8.4% sodium bicarbonate solution (325g) to stir and wash, and separate the liquids; stir and wash the organic phase with 20% sodium sulfite solution (350g), and separate the liquids; stir and wash the organic phase with 20% sodium chloride solution (350g), concentrate the organic phase to remove the solvent, and concentrate to dryness as much as possible. Dissolve with tetrahydrofuran (500mL, 10V) to obtain a solution.

[0144] The solution was added to the reactor, and the previously recovered Pd / C was added. The system hydrogen pressure was maintained at 1.0 MPa and the temperature was 55±10°C. The reaction was carried out for 24 hours. The reaction was completed by HPLC monitoring (Compound IVA≤1.0%). The reaction system was filtered with a filter membrane to remove Pd / C. The filtrate was decompressed to remove the solvent, and dichloromethane (150 mL, 3V) and methanol (500 mL, 10V) were added. Dichloromethane was removed under reduced pressure, and the system was maintained at 7-8V until dichloromethane was less than or equal to 3%. A white solid was precipitated during the process. The temperature was lowered for crystallization. The system temperature was maintained at -5-0°C and stirred for 2 hours. The stirring was stopped, and the filter cake was washed with cold methanol (50 mL, 1V), drained, and dried at 50°C with air blast to obtain compound VA42.8g, with a yield of 85.3%, HPLC: 96.4%, and the main impurity was VA-a containing about 2.9%.

[0145] Example 14: Synthesis of Compound VI A

[0146]

[0147] Under nitrogen protection, compound VA (200g, 393mmol) was dissolved in tetrahydrofuran (1.2L, 6V), and 1mol / L lithium tri-tert-butoxyaluminum hydride tetrahydrofuran solution (393mL, 393mmol) was added dropwise at 0±5°C. After the addition was complete, stirring was continued at this temperature for 24 hours. HPLC monitoring confirmed that the reaction was complete (compound VA<3%), and the reaction solution was added dropwise to 2mol / L hydrochloric acid (600mL), the temperature was controlled within 20°C, and the mixture was stirred and separated. The organic phase was separated with saturated sodium chloride (200mL, 1V) to obtain organic phase I and aqueous phase I. The aqueous phase I was extracted with tetrahydrofuran (400mL, 2V) to obtain organic phase II. The organic phases were combined and added with n-heptane (400mL, 2V), and then washed with saturated sodium chloride (400mL, 2V). The organic phase was separated to obtain an organic phase, and then Then, part of the solvent was removed under reduced pressure. When the volume was about 1400 mL (7V), water was removed by splitting if any. n-heptane (2.0 L, 10V) was added and stirred. The organic phase was further distilled under reduced pressure to remove tetrahydrofuran (judged by the consistency of the density of the evaporated liquid with that of n-heptane). At the same time, the volume of the system was ensured to be around 2000 mL (10V). The solution was stirred at 15±10°C for 2 hours, filtered, and the filter cake was washed with 200 mL (1V) of n-heptane and dried to obtain 172.6 g of compound VI A with a yield of 86.0%.

[0148] Example 15: Synthesis of deoxycholic acid (DCA)

[0149]

[0150] Compound IVA (102.1 g, 200 mmol) was dissolved in tetrahydrofuran (715 mL, 7V) with stirring, and 4 mol / L sodium hydroxide solution (200 mL, 800 mmol) was added dropwise at room temperature, and then stirred at 60±5°C for 36 hours. After the reaction was completed by HPLC monitoring, 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 2042 mL (20V), washed with ethyl acetate (200 mL*2), the pH was adjusted to 1-2 with about 7% hydrochloric acid, stirred at 70±10°C for 2 hours, cooled to 20±10°C, filtered, the filter cake was washed with water (510 mL, 5V) 2-3 times, and dried to obtain a solid, which was dissolved in methyl tert-butyl ether (1020 mL, 10V) at 20±5°C, stirred for about 12 hours, filtered, the filter cake was washed with methyl tert-butyl ether (408 mL, 4V), and dried to obtain 72.2 g of deoxycholic acid with a yield of 92.0%.

[0151] Example 16: Synthesis of Compound IIIB

[0152]

[0153] Compound IIB (50 g, 116.7 mmol), tetrahydrofuran (500 mL, 10 V) and 10% Pd / C (50% wet, 2.5 g, 5% wt) were added to the reactor, and nitrogen and hydrogen replacement were performed in sequence. Then, the reaction was carried out at 20±5°C and a hydrogen pressure of 1.5 MPa for 24 hours. HPLC monitoring showed that the reaction was complete. After the reactor was replaced with nitrogen, the reaction liquid was filtered, the filter cake was washed with tetrahydrofuran (50 mL, 1 V), and the filtrate was collected and the solvent was removed under reduced pressure to obtain a white solid compound IIIB.

[0154] 48.2 g, yield 96.0%.

[0155] Example 17: Synthesis of Compound IVB

[0156]

[0157] Step 1:

[0158] Compound IIIB (43.1 g, 100 mmol) and acetonitrile (430 mL, 10 V) were added to the reaction kettle, and cuprous iodide (11.4 g, 60 mmol) and 70% TBHP aqueous solution (51.5 g, 400 mmol) were added in sequence at 45-50° C., and the mixture was stirred for 8 hours. The reaction was completed by monitoring by HPLC (compound IIIB <2%). The temperature of the reaction system was lowered to 20±5° C., and ethyl acetate (5.0 L, 5 V) and saturated sodium sulfite solution (215 mL, 5 V) were then added, stirred, filtered, and saturated sodium chloride solution (129 mL, 3 V) was added to the filtrate, stirred, separated, and the organic phase was decompressed to remove the solvent to obtain 36.6 g of a mixture of oily substances IVB and IVB-a. HPLC showed that the percentage ratio of the compounds was: IIIB / IVB / IVB-a=1.2 / 69.5 / 23.6.

[0159] Step 2:

[0160] The mixture of step 1 (18.3 g, containing about 4.3 g of IVB-a, 9.7 mmol) was added to the reactor, and N,N-dimethylformamide (91.5 mL, 5 V) was added. 2-iodoacylbenzoic acid (5.4 g, about 19.4 mmol, based on 2.0 eq of IVB-a) was added at 20±10°C, and then reacted at 20±10°C for 4 hours. HPLC monitoring confirmed that the reaction was complete (IVB-a <2%, the percentage of IIIB / IVB / IVB-a was 1.1 / 95.1 / 0.0). Ethyl acetate (183 mL, 10 V) and water (183 mL, 10 V) were added to the reaction solution, stirred, and filtered with a filter membrane. The filtrate obtained was separated to obtain an organic phase I. The filter cake was rinsed with ethyl acetate (36.6 mL, 2 V), and the rinsed ethyl acetate was mixed with the water phase, stirred, and separated to obtain an organic phase II, and the organic phases were combined. Use 8.4% sodium bicarbonate solution (70g) to stir and wash, and separate the liquids; use 20% sodium sulfite solution (75g) to stir and wash the organic phase, and separate the liquids; the organic phase is then stirred and washed with 20% sodium chloride solution (75g), the organic phase is concentrated to remove the solvent, methanol (137.2mL, 7.5V) is added, heated to 60±5℃, stirred and dissolved, pure water (45.7mL, 2.5V) is added dropwise, the temperature is lowered to 10±5℃, stirred for 2 hours, filtered, the solid is rinsed with 30% v / v methanol aqueous solution (18.3mL, 1V), dried, and vacuum dried at 45±5℃ to obtain the compound

[0161] IVB 17.9 g, the total yield of two steps is about 80.6%.

[0162] Example 18: Synthesis of Compound IVB

[0163] Step 1: Same as Example 17.

[0164] Step 2:

[0165] The product of step 1 in Example 17 (18.3 g, containing about 4.3 g of IVB-a, 9.7 mmol) was added to a reaction bottle, acetonitrile (183 mL, 10 V) was added, 2-iodoacylbenzoic acid (8.1 g, 29.0 mmol, about 3.0 eq) was added at 50±10° C., and the reaction was carried out at 75±5° C. for 24 hours. HPLC monitoring showed that the percentage ratio of IIIB / IVB / IVB-a was 1.0 / 94.1 / 0.9, and the reaction endpoint was reached (

[0166] IVB-a<2%). Ethyl acetate (183 mL, 10V) and water (183 mL, 10V) were added, stirred, and filtered with a filter membrane to obtain a filtrate, and separated to obtain an organic phase I. The filter cake was rinsed with ethyl acetate (36.6 mL, 2V), and the rinsed ethyl acetate was mixed with the water phase, stirred, and separated to obtain an organic phase II. The organic phases were combined. 8.4% sodium bicarbonate solution (70 g) was used for stirring and washing, and the liquid was separated; the organic phase was stirred and washed with 20% sodium sulfite solution (75 g), and the liquid was separated; the organic phase was stirred and washed with 20% sodium chloride solution (75 g), the organic phase was concentrated to remove the solvent, methanol (137.2 mL, 7.5 V) was added, heated to 60±5° C. and stirred to dissolve, pure water (45.7 mL, 2.5 V) was added dropwise, the temperature was lowered to 10±5° C. and stirred for 2 hours, filtered, and the solid was rinsed with 30% v / v methanol aqueous solution (18.3 mL, 1 V), dried by suction, and dried in vacuo at 45±5° C. to obtain 17.7 g of compound IVB, with a total yield of about 79.7% in two steps.

[0167] Example 19: Synthesis of Compound VB

[0168]

[0169] A solution of compound IVB (25.0 g, 56.2 mmol) in tetrahydrofuran (250 mL, 10 V) and 10% Pb / C (dry, 5.0 g, 20% wt) were added to a reaction kettle, and nitrogen and hydrogen replacement were performed successively. The mixture was stirred for 24 hours at a pressure of 1.3 MPa and a temperature of 55±10° C., and the completion of the reaction was determined by HPLC monitoring (compound IVB≤3%, and the percentage ratios of compound IVB / compound VB / enol compound VB-a / hydroxy compound VB-b were 0.02 / 0.69 / 0.22 / 0.05).

[0170] The reaction solution was filtered, and the filtrate was decompressed to remove the solvent (containing VB-a and VB-b of 12.6 mmol and 2.8 mmol, respectively, about 15.4 mmol in total), and N,N-dimethylformamide (125 mL, 5V) was added, and 2-iodoacylbenzoic acid (12.9 g, about 46.2 mmol, 3.0 eq based on the total of VB-a and VB-b) was added at 25±5°C for 4 hours. HPLC monitoring confirmed that the reaction was complete (VB-a and VB-b were <1.0%). Ethyl acetate (250 mL, 10V) and water (250 mL, 10V) were added to filter, stirred, and filtered with a filter membrane. The obtained filtrate was separated, and the filter cake was rinsed with ethyl acetate (50 mL, 2V). The rinse solution was mixed with the aqueous phase, stirred, extracted, separated, and the organic phase was combined. Use 8.4% sodium bicarbonate solution (160g) to stir and wash, and separate the liquids; use 20% sodium sulfite solution (165g) to stir and wash the organic phase, and separate the liquids; the organic phase is then stirred and washed with 20% sodium chloride solution (165g), and the organic phase is concentrated to remove the solvent and concentrated to dryness as much as possible. Dissolve with tetrahydrofuran (250mL, 10V) to obtain a solution.

[0171] The solution was added to a reactor, and 10% / C (dry, 2.5 g, 10% wt) was added. The reaction was carried out at 55±10°C and a hydrogen pressure of 1.3 MPa for 24 hours. The reaction was completed by HPLC monitoring (Compound IVB≤1.0%). The reaction solution was filtered to remove Pd / C, the filtrate was decompressed to remove the solvent, methanol (150 mL, 6V) was added, heated to dissolve, pure water (75 mL, 3V) was added dropwise, the temperature was lowered for crystallization, stirred at 15±5°C for 2-4 hours, filtered, the filter cake was washed with 50% v / v methanol aqueous solution (25 mL, 1V), drained, and air-dried at 50°C to obtain 19.1 g of compound VB with a yield of 76.4%.

[0172] Example 20: Synthesis of Compound VIB

[0173]

[0174] Under nitrogen protection, compound VB (20 g, 44.8 mmol) was dissolved in tetrahydrofuran (160 mL, 8 V), and 1 mol / L lithium tri-tert-butoxyaluminum hydride tetrahydrofuran solution (67.2 mL, 67.2 mmol, 1.5 eq) was added dropwise at 0±5°C. After the addition was complete, stirring was continued at this temperature for 24 hours. HPLC monitoring confirmed that the reaction was complete (compound VB <3%). The reaction solution was cooled and 2 mol / L hydrochloric acid solution (70 mL) was added dropwise at a temperature not higher than 10°C. The solution was separated and the organic phase was washed with saturated sodium chloride (20 mL, 1V) and 2 mol / L hydrochloric acid solution (20 mL, 1V) in sequence with stirring. The solution was separated and the organic phase was washed with saturated sodium chloride (40 mL, 2V). Then, part of the solvent was removed under reduced pressure. When the volume was about 40 mL, n-heptane (200 mL, 10V) was added with stirring. The solution was separated and tetrahydrofuran was removed from the organic phase by distillation under reduced pressure (determined by the consistency of the density of the evaporated liquid with that of n-heptane). The volume of the system was about 100 mL. The solution was stirred at 10-25°C for 2 hours and filtered. The filter cake was washed with a small amount of n-heptane and dried to obtain 15.8 g of compound VIB with a yield of 78.6%.

[0175] Example 21: Synthesis of deoxycholic acid

[0176]

[0177] Compound VIB (15.0 g, 33.5 mmol) was dissolved in tetrahydrofuran (150 mL, 10 V) by stirring, and 4 mol / L sodium hydroxide solution (33.5 mL, 134.0 mmol) was added dropwise at room temperature, and the mixture was stirred and reacted at 60±5° C. for 24 hours. After the reaction was completed by HPLC monitoring, tetrahydrofuran was removed under reduced pressure, water (120 mL, 8 V) was added to the residue, 1 mol / L hydrochloric acid was added dropwise at 10-15° C. to adjust the pH to 9-10, and the organic solvent was removed under reduced pressure. After the residue was cooled, it was washed with ethyl acetate (30 mL*2), and the pH was adjusted to 1-2 with 1 mol / L hydrochloric acid, stirred for 2 hours, filtered, and the filter cake was washed with water (75 mL, 5 V), and air-dried to obtain deoxycholic acid solid (12.2 g, 84.0%).

Claims

1. A method for preparing a deoxycholic acid intermediate compound IV, comprising subjecting compound IV-a to an oxidation reaction to obtain compound IV; The reaction formula is as follows: in, P is benzoyl or acetyl; the oxidant of the oxidation reaction is selected from 2-iodoacylbenzoic acid and Dess-Martin reagent; the molar ratio of the oxidant to compound IV-a is (1-3):1; the reaction solvent is selected from dimethyl sulfoxide, dimethylformamide and acetonitrile.

2. The method according to claim 1, wherein P is a benzoyl group, the oxidant of the oxidation reaction is selected from 2-iodoacylbenzoic acid; the molar ratio of the oxidant to compound IV-a is (1.5-2):1; and the reaction solvent is selected from dimethyl sulfoxide and dimethylformamide.

3. A method for preparing deoxycholic acid, comprising the following steps: 1) Compound II is subjected to hydrogenation reaction to obtain compound III; 2) Compound III is subjected to a first oxidation reaction to obtain a mixture containing Compound IV and Compound IV-a; the mixture is subjected to a second oxidation reaction to obtain Compound IV; 3) Compound IV is subjected to a first hydrogenation reaction to obtain a mixture containing compounds V, Va and Vb; the mixture is subjected to an oxidation reaction and a second hydrogenation reaction in sequence to obtain compound V; 4) Compound V is subjected to reduction reaction to obtain compound VI; 5) Compound VI is subjected to deprotection and hydrolysis to obtain deoxycholic acid I or its salt; The reaction formula is as follows: Wherein: P is benzoyl or acetyl; In step 1), the hydrogenation reaction is carried out in the presence of a Pd / C or PtO2 catalyst; In step 2), the first oxidation reaction is carried out in the presence of an oxidant and a co-oxidant; the oxidant is tert-butyl hydroperoxide; the co-oxidant is CuI; In step 2), the oxidant used in the second oxidation reaction is selected from 2-iodoacylbenzoic acid and Dess-Martin periodinane; the molar ratio of the oxidant to compound IV-a is (1-3):1; the reaction solvent is selected from dimethyl sulfoxide, dimethylformamide and acetonitrile; In step 3), the first hydrogenation reaction and the second hydrogenation reaction are carried out in the presence of a Pd / C or PtO2 catalyst; the oxidant in the oxidation reaction is selected from Dess-Martin oxidant and 2-iodoacylbenzoic acid; the molar ratio of the oxidant to the sum of compounds V-a and V-b is (1-3):1; In step 4), the reduction reaction is carried out in the presence of a reducing agent, lithium tri-tert-butoxyaluminum hydride; In step 5), the reaction conditions include alkali metal hydroxide, alkali metal alkoxide or a mixture thereof.

4. The method of claim 3, wherein in step 2), the oxidant used in the second oxidation reaction is 2-iodoacylbenzoic acid; the molar ratio of the oxidant to compound IV-a is (1.5-2):1; the reaction solvent is selected from dimethyl sulfoxide and dimethylformamide; in step 3), the first hydrogenation reaction and the second hydrogenation reaction are carried out in the presence of a Pd / C catalyst; the oxidant of the oxidation reaction is 2-iodoacylbenzoic acid.

5. A method for preparing a deoxycholic acid intermediate compound IV, comprising the following steps: 1) Compound II is subjected to hydrogenation reaction to obtain compound III; 2) Compound III is subjected to a first oxidation reaction to obtain a mixture containing Compound IV and Compound IV-a; the mixture is subjected to a second oxidation reaction to obtain Compound IV; Wherein: P is benzoyl or acetyl; In step 1), the hydrogenation reaction is carried out in the presence of a Pd / C or PtO2 catalyst; In step 2), the first oxidation reaction is carried out in the presence of an oxidant and a co-oxidant; the oxidant is tert-butyl hydroperoxide; the co-oxidant is CuI; In step 2), the oxidant used in the second oxidation reaction is selected from 2-iodoacylbenzoic acid and Dess-Martin periodinane; the molar ratio of the oxidant to compound IV-a is (1-3):1; and the reaction solvent is selected from dimethyl sulfoxide, dimethylformamide and acetonitrile.

6. The method of claim 5, wherein P is benzoyl, the hydrogenation reaction in step 1) is carried out in the presence of a Pd / C catalyst; in step 2), the oxidant used in the second oxidation reaction is 2-iodoacylbenzoic acid; the molar ratio of the oxidant to compound IV-a is (1.5-2):1; and the reaction solvent is selected from dimethyl sulfoxide and dimethylformamide.

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