WO2020177240A1 - 鹅去氧胆酸及其制备方法 - Google Patents

鹅去氧胆酸及其制备方法 Download PDF

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WO2020177240A1
WO2020177240A1 PCT/CN2019/091965 CN2019091965W WO2020177240A1 WO 2020177240 A1 WO2020177240 A1 WO 2020177240A1 CN 2019091965 W CN2019091965 W CN 2019091965W WO 2020177240 A1 WO2020177240 A1 WO 2020177240A1
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reaction
acid
chenodeoxycholic acid
preparation
molar ratio
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French (fr)
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张雷
黄欢
李杉
张和平
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Zhongshan Bailing Biotechnology Co Ltd
South China University of Technology SCUT
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Zhongshan Bailing Biotechnology Co Ltd
South China University of Technology SCUT
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane

Definitions

  • This application relates to the field of drug synthesis, specifically, to chenodeoxycholic acid and a preparation method thereof.
  • Bile acids are important endogenous molecules related to countless biological functions, including the absorption and excretion of cholesterol, and play an important role in fat metabolism.
  • Chenodeoxycholic acid is a natural primary bile acid, which is widely present in the bile of humans, livestock and poultry, and is the main component in the bile of poultry such as chickens, ducks and geese. Since Thistle and Schoenfirld discovered that chenodeoxycholic acid can treat gallstones, the clinical application of chenodeoxycholic acid has been continuously discovered. Since the 1970s, chenodeoxycholic acid has been mainly used for the treatment of gallstone diseases and other liver and gallbladder diseases.
  • chenodeoxycholic acid has antibacterial, anti-inflammatory and anti-asthmatic effects in addition to various liver and gallbladder diseases. Antitussive, promotion of digestion in the digestive system, treatment of tendon xanthomas and insulin resistance and other medicinal values, at the same time, chenodeoxycholic acid is also an important raw material for the synthesis of ursodeoxycholic acid and obeticholic acid. Therefore, with the expansion of medicinal value and the expansion of demand for ursodeoxycholic acid and obeticholic acid, the demand for chenodeoxycholic acid is also increasing.
  • the current chemical synthesis pathways mainly use cholic acid and hyodeoxycholic acid as raw materials for synthesis.
  • cholic acid There are two ways to synthesize with cholic acid as raw materials.
  • the yield can reach 40%.
  • the current yield of the synthetic route using hyodeoxycholic acid as raw material is 26%, but this route uses chloranil, which has a greater impact on the environment.
  • This application provides a method for preparing chenodeoxycholic acid, which can use extraction by-products as raw materials to rapidly synthesize chenodeoxycholic acid, with mild conditions in each step of the route and high yield, which is suitable for mass production.
  • the application also provides a chenodeoxycholic acid, which is prepared by the above method.
  • a preparation method of chenodeoxycholic acid includes the following steps:
  • the intermediate E is formed after a chemical reaction, and then the intermediate E undergoes a chemical reaction to form chenodeoxycholic acid;
  • the structural formula of the intermediate E is as follows: Among them, R1 is an alkyl group, an alkenyl group or an aromatic group, and R2 is an acyl group.
  • a chenodeoxycholic acid prepared by the above-mentioned preparation method of chenodeoxycholic acid.
  • this application uses duck gall, goose gall and other by-products obtained after extraction as reaction raw materials, which can achieve waste utilization, reduce synthesis costs, and have a wide range of raw materials.
  • the yield of chenodeoxycholic acid obtained through the above steps is high, up to 32%, which is suitable for mass production.
  • the preparation method has simple operation, strong reproducibility, and extremely strong practicability.
  • Fig. 1 is a synthetic route diagram of chenodeoxycholic acid provided in an embodiment of the application.
  • a method for preparing chenodeoxycholic acid includes the following steps:
  • 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholanoic acid (abbreviation: allochodeoxycholic acid) (A) is used as a raw material to obtain 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholanoic acid ester ( B), 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholanoic acid ester (B) is selectively oxidized at position 3 to obtain 3-keto-7 ⁇ -hydroxy-5 ⁇ -cholanoic acid ester (C), 3-keto-7 ⁇ -hydroxy -5 ⁇ -cholanoic acid ester (C) is protected at the 7 position to obtain 3-keto-7 ⁇ -acyloxy-5 ⁇ -cholanoic acid ester (D), 3-keto-7 ⁇ -acyloxy-5 ⁇ -cholanoic acid The ester (D) undergoes oxidation reaction to obtain ⁇ 1,4-3-one-7 ⁇ -acyloxy-unsaturated cholanoic acid ester (E), ⁇ 1,4-3-one-7 ⁇ -acyloxy-unsatur
  • the raw material used is 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholanoic acid, which is a by-product obtained by extraction of duck gall and goose gall.
  • goose Oxide can be quickly prepared Cholic acid; using waste as a raw material, a wide range of sources, sufficient supply can also reduce synthesis costs.
  • the yield of chenodeoxycholic acid obtained through the above steps is high, up to 32%, which is suitable for mass production.
  • R1 of the intermediate E is an alkyl group, an alkenyl group or an aromatic group
  • R2 is an acyl group.
  • the intermediate E is an intermediate obtained by sequentially subjecting the 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholanoic acid to the first esterification reaction, oxidation reaction, second esterification reaction and dehydrogenation reaction.
  • the first esterification reaction is the esterification reaction of 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholanoic acid with alcohol.
  • the 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholanoic acid Intermediate B can be obtained by reacting with alcohol at 50-90°C; the temperature is preferably 60-70°C, and the reaction temperature can be within the above range, and can be adjusted according to actual needs.
  • the reaction temperature can be 60°C, 61 °C, 62°C, 63°C, 64°C, 65°C, 66°C, 67°C, 68°C, 69°C, 70°C and so on.
  • the reaction time is 2-6h.
  • the reaction time can be adjusted according to the progress of the esterification reaction to complete the esterification reaction, such as 2h, 2.5h, 3h, 3.5h, 4h, 4.5h, 5h, 5.5h, 6h, etc. .
  • the catalyst is an acidic substance, more preferably any one of concentrated hydrochloric acid, concentrated sulfuric acid, benzenesulfonic acid, methanesulfonic acid, and p-toluenesulfonic acid, and more preferably concentrated hydrochloric acid or concentrated sulfuric acid.
  • the use of the above catalyst can ensure its catalytic effect and the formation of intermediate B.
  • the molar ratio of 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholanoic acid, the alcohol and the catalyst is 1:2-30:0.1-1, which can guarantee 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholesterol within this range
  • the alkanoic acid is smoothly esterified.
  • the alcohol is a monohydric alcohol, more preferably a C1-C10 monohydric alcohol, further preferably methanol, ethanol, isopropanol, propenol and benzyl alcohol, and most preferably methanol or isopropanol.
  • the use of the above alcohol can ensure the yield of chenodeoxycholic acid prepared.
  • the reaction solution is purified, and the purification is to reduce the impurity content in the intermediate B, and then to ensure the purity of the prepared chenodeoxycholic acid and to ensure its yield.
  • the purification method is: after the reaction, the reaction solution obtained from the first esterification reaction is cooled to room temperature, methanol is removed, ethyl acetate is added, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution, dried, and the solvent is removed. , Get Intermediate B.
  • the added quality of ethyl acetate is preferably 10-20 times that of the raw material 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholanoic acid, so as to ensure the purification effect.
  • intermediate B prepared above is reacted with the first oxidant for 2-36 h to form intermediate C, which is an oxidation reaction, which is a 3-position selective oxidation.
  • compound B, the first oxidant, diatomaceous earth, and the first organic solvent are mixed, and reacted under reflux to obtain compound C.
  • the above method ensures that the yield of intermediate C is high.
  • the first oxidant is any one of peroxide, metal compound or bromoimide; more preferably, the peroxide is peroxybenzoic acid or hydrogen peroxide; the metal compound is sodium hypochlorite, silver carbonate , Any one of manganese dioxide or chromium trioxide; the bromoimide is N-bromosuccinimide; most preferably, the first oxidant is sodium hypochlorite, silver carbonate or manganese dioxide.
  • the use of the above-mentioned oxidant can ensure the oxidation effect of the oxidation reaction, thereby ensuring the synthesis rate and yield of intermediate C.
  • the first organic solvent includes toluene, dichloromethane, tetrahydrofuran, chloroform, acetonitrile, preferably toluene.
  • the use of the above-mentioned first organic solvent can provide a good reaction atmosphere for the oxidation reaction and ensure the smooth progress of the reaction.
  • the molar ratio of the intermediate B to the first oxidant is 1:0.5-3, preferably 1:0.8-2, such as 1:1, 1:1.5, 1:2, 1:2.5 in different embodiments , 1:3 and so on.
  • the molar ratio of intermediate B and diatomaceous earth is 1:5-15.
  • it can be 1:6, 1:8, 1:10, 1:12, 1:15, and so on.
  • the mass ratio of the intermediate B and the first organic solvent is 1:5-20. It can be adjusted within this range. Using the above ratio can ensure the smooth progress of the reaction process and ensure the yield of intermediate C.
  • the reaction solution obtained from the oxidation reaction is purified to form Intermediate C.
  • intermediate C is reacted with 4-dimethylaminopyridine, triethylamine, and acid anhydride for 1-15 hours, and then the pH of the reaction solution is adjusted to 6-7 to form intermediate D, which is the second esterification reaction, the second ester
  • the reaction is an esterification reaction at the 7 position.
  • compound C is dissolved in a second organic solvent, 4-dimethylaminopyridine, triethylamine, and acid anhydride are added, and reacted at room temperature for 1-15 hours, and then the reacted reaction solution is added to water to adjust the pH to 6-7. Separate and collect the organic phase to obtain intermediate D.
  • the second organic solvent is ethyl acetate, and the acid anhydride is acetic anhydride.
  • pH adjustment is adjusted by hydrochloric acid with a concentration of 1-3M.
  • the molar ratio of intermediate C, the 4-dimethylaminopyridine, the triethylamine, and the acid anhydride is 1:0.01-0.5:1.5-3:0.8-4.
  • the mass ratio of intermediate C to the second organic solvent is 1:10-15. Using the above ratio can ensure the smooth progress of the second esterification reaction and ensure the yield of intermediate D.
  • the organic phase obtained from the second esterification reaction is purified to form intermediate D.
  • the purification method is: washing the organic phase with a saturated sodium bicarbonate solution and a saturated aqueous salt solution, drying, and removing the solvent to obtain intermediate D .
  • Intermediate D reacts with the second oxidant at 20-90°C for 3-72h, which is a dehydrogenation reaction.
  • the dehydrogenation reaction is a reaction in which positions 1, 2, 4, and 5 are simultaneously dehydrogenated to form olefins.
  • the intermediate D is dissolved in the third organic solvent, the second oxidant and trifluoroacetic acid are added, and the reaction is heated under the protection of nitrogen to obtain the intermediate E.
  • the third organic solvent is preferably dimethyl sulfoxide.
  • the second oxidant is an iodine reagent, preferably a hypervalent iodine reagent, and more preferably 2-iodoylbenzoic acid. The use of the above-mentioned second oxidant and third organic solvent can ensure the production efficiency of the intermediate E.
  • the molar ratio of the intermediate D to the second oxidant is 1:2-5.
  • the molar ratio of intermediate D and trifluoroacetic acid is 1:0.1-0.5, and the mass ratio of intermediate D and the third organic solvent is 1:10-35.
  • the reaction solution of the dehydrogenation reaction is purified by adding the reaction solution after the reaction to water, extracting with ethyl acetate, separating and collecting the organic phase, and washing the organic phase with water and a saturated saline solution. Dry, remove the solvent, and separate by column chromatography to obtain Intermediate E. More preferably, the eluent for column chromatography separation is ethyl acetate and petroleum ether, and the volume ratio of ethyl acetate and petroleum ether is 1:10-20.
  • the use of the above purification method can further ensure the purification effect, and then ensure the purity and yield of chenodeoxycholic acid prepared subsequently.
  • intermediate E is obtained, and then intermediate E is successively subjected to catalytic hydrogenation reduction reaction, 3-position reduction reaction and hydrolysis reaction to obtain the chenodeoxycholic acid.
  • the catalytic hydrogenation reduction reaction is a simultaneous hydrogenation reduction reaction at positions 1, 2, 4, and 5.
  • the intermediate E is mixed with a catalyst and triethylamine and then hydrogenated at 20-60°C.
  • Intermediate F is formed after 0.5-4 hours.
  • intermediate E, catalyst, alcohol solvent and triethylamine are reacted, and the reaction pressure is 0.8-1.2 MPa.
  • the catalyst is palladium on carbon, preferably 5% palladium on carbon, and the alcohol solvent is methanol.
  • the molar ratio of the intermediate E to the catalyst and triethylamine is 1:2-10:0.001-0.5; the mass ratio of the intermediate E to the alcohol solvent is 1:10-20. Using the above ratio can ensure the smooth progress of the catalytic hydrogenation reduction reaction and ensure the yield of intermediate F.
  • the reaction solution obtained by the catalytic hydrogenation reduction reaction is purified.
  • the alcohol solvent is spin-dried, dissolved with ethyl acetate, washed with hydrochloric acid and saturated brine, and the solvent is removed to obtain Intermediate F.
  • the added quality of ethyl acetate is preferably 20-30 times that of intermediate F.
  • wash use similar volumes of 1-3M hydrochloric acid and saturated brine to wash, preferably the same volume.
  • the 3-position reduction reaction is to react the intermediate F with a reducing agent.
  • the 3-position reduction reaction means that the 3-position carbonyl group undergoes a catalytic hydrogenation reaction to obtain a hydroxyl group.
  • the intermediate F is dissolved in the fourth organic solvent, a reducing agent is added at 0-10° C., and the reaction is performed at room temperature to obtain the intermediate G.
  • the reducing agent is a hydride, more preferably sodium borohydride.
  • the fourth organic solvent is preferably anhydrous methanol.
  • the reaction time is monitored by thin layer chromatography TLC, and the developing agent is ethyl acetate and petroleum ether in a volume ratio of 1:6-1:2. The reaction in the above manner can ensure the smooth progress of the reaction and the yield of the prepared intermediate G.
  • the molar ratio of intermediate F to reducing agent is 1:0.5-10, and the mass ratio of intermediate F to the fourth organic solvent is 1:30-50. Using the above ratio can further ensure the preparation effect of intermediate G.
  • the reaction solution obtained by the 3-position reduction reaction is purified.
  • the purification method is: add water and ethyl acetate to the reaction solution after the reaction, fully stir, stand still for layering, collect the organic phase, and then use saturated Wash with sodium bicarbonate and saturated brine, dry, and remove the solvent to obtain Intermediate F.
  • the masses of the added water and ethyl acetate are preferably 5-10 times and 10-30 times the mass of the reaction solution, respectively.
  • the intermediate G is hydrolyzed under alkaline conditions to obtain chenodeoxycholic acid, which is 3 ⁇ ,7 ⁇ -dihydroxy-5 ⁇ -cholanoic acid.
  • the intermediate G, alcohol solvent, water, and alkali metal hydroxide are mixed, reacted under reflux, concentrated to remove the alcohol solvent, water and hydrochloric acid are added, the pH is adjusted to 3-4, the precipitate is precipitated, and the precipitate is collected by filtration to obtain a goose Deoxycholic acid. More preferably, the reaction is carried out at 60-80°C for 4-8 hours.
  • the alcohol solvent is methanol
  • the alkali metal hydroxides include sodium hydroxide and potassium hydroxide, and sodium hydroxide is preferred. Using the above preparation process ensures production efficiency and purity.
  • the mass ratio of intermediate G, alcohol solvent and water is 1:10-20:5-10.
  • the molar ratio of intermediate G to alkali metal hydroxide is 1:2-6. The use of the above ratio can ensure the full progress of the reaction and reduce the generation of by-products.
  • the amount of water added is preferably the same volume as the reaction liquid.
  • the concentration of the added hydrochloric acid is 1-3M.
  • the application also provides a chenodeoxycholic acid, which is prepared by the above-mentioned preparation method of chenodeoxycholic acid.
  • reaction conditions of each step of the present application are relatively mild, and the process is simple.
  • the product is obtained through 7-step reaction, which is easy to control, and the target compound can be obtained without complicated equipment.
  • This embodiment provides a method for preparing chenodeoxycholic acid, including the following steps:
  • This embodiment also provides a chenodeoxycholic acid, which is prepared by the above preparation method.
  • the total yield was 32 and the purity was 98.7%.
  • Example 2- The preparation method of chenodeoxycholic acid provided in Example 10 is basically similar to the preparation method of chenodeoxycholic acid provided in Example 1, except that the raw materials used and the specific conditions of each step are different.
  • the total yield is 30% and the purity is 97%.
  • the total yield is 28% and the purity is 98%.
  • reaction temperature is 50°C
  • reaction time is 36h
  • the molar ratio of intermediate D and 2-iodoylbenzoic acid is 1:3
  • the molar ratio of intermediate D and trifluoroacetic acid is 1:0.2
  • the mass ratio of the intermediate D and the third organic solvent is 1:11.
  • the volume ratio of ethyl acetate and petroleum ether during purification is 1:15.
  • the total yield is 28% and the purity is 99%.
  • reaction temperature is 20°C
  • reaction time is 12h
  • the molar ratio of intermediate D and 2-iodoylbenzoic acid is 1:5
  • the molar ratio of intermediate D and trifluoroacetic acid is 1:0.1
  • the mass ratio of intermediate D and the third organic solvent is 1:15.
  • the volume ratio of ethyl acetate and petroleum ether during purification is 1:10.
  • the total yield is 26% and the purity is 98%.
  • Example 1 The difference between the preparation method of this example and Example 1 is that when preparing intermediate D, the amount of acetic anhydride used is 0.98 mmol. The total yield is 30% and the purity is 99%.
  • Example 1 The difference between the preparation method of this example and Example 1 is that when preparing Intermediate D, the amount of acetic anhydride used is 1.72 mmol. The total yield is 31% and the purity is 98%.
  • Example 1 The difference between the preparation method of this example and Example 1 is that when preparing Intermediate E, the amount of 2-iodoylbenzoic acid used is 4.48 mmol. The total yield is 28% and the purity is 98%.
  • Example 1 The difference between the preparation method of this example and Example 1 is that when preparing Intermediate G, the amount of sodium borohydride used is 0.148 mmol. The total yield is 29% and the purity is 98%.
  • Example 1 The difference between the preparation method of this example and Example 1 is that the amount of sodium borohydride used in the preparation of Intermediate G is 0.74 mmol. The total yield is 31% and the purity is 98%.
  • the present application uses duck gall, goose gall and other by-products discarded after extraction as reaction raw materials, which can achieve waste utilization, reduce synthesis costs, and have a wide range of raw materials and sufficient supply.
  • the yield of chenodeoxycholic acid obtained through the above steps is high, up to 32%, which is suitable for mass production.
  • the preparation method has simple operation, strong reproducibility, and extremely strong practicability.

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Abstract

本申请涉及药物合成领域，具体而言，涉及一种鹅去氧胆酸及其制备方法。鹅去氧胆酸的制备方法，包括以下步骤：利用3α，7α-二羟基-5α-胆烷酸为原料经过化学反应后形成中间体E，而后中间体E再经过化学反应形成鹅去氧胆酸；所述中间体E的结构式如下：其中，R1为烷基、烯基或者芳香基团，R2为酰基。该制备方法能够利用提取副产物作为原料，快速合成鹅去氧胆酸，路线各步骤条件温和，收率高，适合大量制备。

Description

鹅去氧胆酸及其制备方法
本申请是以申请号为201910167593.3、申请日为2019年3月6日的中国专利申请为基础，并主张其优先权，该申请的全部内容在此作为整体引入本申请中。
技术领域
本申请涉及药物合成领域，具体而言，涉及一种鹅去氧胆酸及其制备方法。
背景技术
胆汁酸是与无数生物学功能相关的重要内源性分子，包括胆固醇的吸收和排泄，在脂肪代谢中起到重要作用。鹅去氧胆酸是天然的初级胆汁酸，在人、畜、禽的胆汁中广泛存在，是鸡、鸭、鹅等家禽胆汁中主要成分。自从Thistle和Schoenfirld发现鹅去氧胆酸能治疗胆结石之后，鹅去氧胆酸在临床上的应用就不断被发现。20世纪70年代以来，鹅去氧胆酸主要用于胆结石疾病和其他肝胆疾病的治疗，随后，陆续发现鹅去氧胆酸除了能治疗各种肝胆疾病外，还具有抗菌、消炎、平喘镇咳、促进消化系统的消化、治疗脑腱性黄瘤病和胰岛素耐受性等药用价值，同时，鹅去氧胆酸还是合成熊去氧胆酸和奥贝胆酸的重要原料。因此，随着药用价值的扩大以及对熊去氧胆酸和奥贝胆酸需求的扩大，鹅去氧胆酸的需求也越来越大。
目前的化学合成途径主要是以胆酸和猪去氧胆酸为原料进行合成。以胆酸为原料进行的合成有两条途径，一是通过还原12位的羰基制备鹅去氧胆酸；二是通过C11烯烃加氢制备鹅去氧胆酸，这两条途径经过工艺优化后收率可以达到40％。以猪去氧胆酸为原料的合成 途径目前的收率在26％，但是这条路线使用到了四氯苯醌，对环境的影响较大。
申请内容
本申请提供了一种鹅去氧胆酸的制备方法，其能够利用提取副产物作为原料，快速合成鹅去氧胆酸，路线各步骤条件温和，收率高，适合大量制备。
本申请还提供一种鹅去氧胆酸，其通过上述方法制备得到。
本申请是这样实现的：
一种鹅去氧胆酸的制备方法，包括以下步骤：
利用3α，7α-二羟基-5α-胆烷酸为原料经过化学反应后形成中间体E，而后中间体E再经过化学反应形成鹅去氧胆酸；所述中间体E的结构式如下：
Figure PCTCN2019091965-appb-000001
其中，R1为烷基、烯基或者芳香基团，R2为酰基。
一种鹅去氧胆酸，其通过上述鹅去氧胆酸的制备方法制备得到。
本申请的有益效果是：本申请通过采用鸭胆、鹅胆等经过提取后得到的副产物作为反应原料，能够做到废物利用，降低合成成本，且原料来源广泛。并且，经过上述步骤得到的鹅去氧胆酸的产率高，可达32％，适合大量制备。且该制备方法操作简单，重复性强，具有极强的实用性。
附图说明
为了更清楚地说明本申请具体实施方式或现有技术中的技术方案，下面将对具体实施方式或现有技术描述中所需要使用的附图作简 单地介绍，显而易见地，下面描述中的附图是本申请的一些实施方式，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1为本申请实施例提供的鹅去氧胆酸的合成路线图。
具体实施方式
为使本申请实施例的目的、技术方案和优点更加清楚，下面将对本申请实施例中的技术方案进行清楚、完整地描述。实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市售购买获得的常规产品。
下面对本申请实施例的鹅去氧胆酸及其制备方法进行具体说明。
一种鹅去氧胆酸的制备方法，参见图1，包括以下步骤：
按照下式进行化学反应：
Figure PCTCN2019091965-appb-000002
本申请实施例以3α，7α-二羟基-5α-胆烷酸(简称：别鹅去氧胆酸)(A)为原料经酯化得到3α,7α-二羟基-5α-胆烷酸酯(B)，3α,7α-二羟基-5α-胆烷酸酯(B)3位选择性氧化得到3-酮-7α-羟基-5α-胆烷酸酯(C)，3-酮-7α-羟基-5α-胆烷酸酯(C)的7 位保护得到3-酮-7α-酰氧基-5α-胆烷酸酯(D)，3-酮-7α-酰氧基-5α-胆烷酸酯(D)进行氧化反应得到△1,4-3-酮-7α-酰氧基-不饱和胆烷酸酯(E)，△1,4-3-酮-7α-酰氧基-不饱和胆烷酸酯(E)进行催化氢化还原双键得到3-酮-7α-酰氧基-5β-胆烷酸酯(F)，3-酮-7α-酰氧基-5β-胆烷酸酯(F)的3位还原得到3α-羟基-7α-酰氧基-5β-胆烷酸酯(G)，3α-羟基-7α-酰氧基-5β-胆烷酸酯(G)水解脱保护得到鹅去氧胆酸(H)。
采用的原料为3α，7α-二羟基-5α-胆烷酸，其为鸭胆、鹅胆等经过提取得到的副产物，采用上述丢弃不用的副产物作为原料，能够快速的制备得到鹅去氧胆酸；以废物作为原料，来源广泛，供应充足还能够降低合成成本。并且，经过上述步骤得到的鹅去氧胆酸产率高，可达32％，适合大量制备。
上式步骤中的中间体E的R1为烷基、烯基或者芳香基团，R2为酰基。而中间体E是将所述3α，7α-二羟基-5α-胆烷酸依次经过第一次酯化反应、氧化反应、第二次酯化反应以及脱氢反应后得到的中间体。
S1、制备中间体B；
第一次酯化反应为3α，7α-二羟基-5α-胆烷酸与醇进行的酯化反应，具体是在催化剂的作用下，将所述3α，7α-二羟基-5α-胆烷酸与醇在50-90℃的条件下，反应得到中间体B；温度优选为60-70℃，反应温度在上述范围内均可，可根据实际需求进行调整，如反应温度可以为60℃、61℃、62℃、63℃、64℃、65℃、66℃、67℃、68℃、69℃、70℃等等。
反应时间为2-6h，可以可根据酯化反应进度调节反应时间，使酯化反应完全，如可以为2h、2.5h、3h、3.5h、4h、4.5h、5h、5.5h、6h等等。
催化剂为酸性物质，更优选为浓盐酸、浓硫酸、苯磺酸、甲磺酸、对甲苯磺酸中的任一种，更优选的，为浓盐酸或浓硫酸。采用上述催化剂能够保证其催化效果，保证中间体B的形成。
3α，7α-二羟基-5α-胆烷酸、所述醇和所述催化剂的摩尔比为1:2-30：0.1-1，在这一范围内能够保证3α，7α-二羟基-5α-胆烷酸顺利酯化，如在不同实施方式中，3α，7α-二羟基-5α-胆烷酸和醇的摩尔比可以为1﹕2、1﹕4、1﹕6、1﹕8、1﹕10、1﹕12、1﹕14、1﹕16、1﹕18、1﹕20、1﹕25、1﹕30等等；3α，7α-二羟基-5α-胆烷酸和催化剂的摩尔比可以为1﹕0.1、1﹕0.3、1﹕0.5、1﹕0.7、1﹕0.9、1﹕1等等。
进一步地，醇为一元醇，更优选为C1-C10一元醇，进一步优选为甲醇、乙醇、异丙醇、丙烯醇和苄醇，最优选为，甲醇或者异丙醇。采用上述醇能够保证制备得到的鹅去氧胆酸的产率。
反应结束后对反应液进行提纯，提纯是为了降低中间体B中的杂质含量，继而保证制备得到的鹅去氧胆酸的纯度，且保证其产率。
提纯方法是：反应结束后，将第一次酯化反应得到的反应液冷却至室温，除去甲醇，加入乙酸乙酯，依次用饱和碳酸氢钠溶液和饱和氯化钠溶液洗涤，干燥，除去溶剂，得到中间体B。其中，乙酸乙酯的加入质量优选为原料3α，7α-二羟基-5α-胆烷酸的10-20倍，从而保证提纯效果。
S2、制备中间体C；
将上述制备得到的中间体B与第一氧化剂反应2-36h形成中间体C，也就是氧化反应，该氧化反应为3位选择性氧化。具体地，化合物B、第一氧化剂、硅藻土和第一有机溶剂混合，回流反应，得到化合物C。采用上述方法保证制备得到中间体C的收率高。
进一步地，第一氧化剂为过氧化物、金属化合物或者溴代酰亚胺中的任意一种；更优选，所述过氧化物为过氧苯甲酸或者双氧水；所述金属化合物为次氯酸钠、碳酸银、二氧化锰或者三氧化铬中的任意一种；所述溴代酰亚胺为N-溴代丁二酰亚胺；最优选，所述第一氧化剂为次氯酸钠、碳酸银或者二氧化锰。采用上述氧化剂，能保证氧化反应的氧化效果，从而保证中间体C的合成速率和产率。
进一步地，第一有机溶剂包括甲苯、二氯甲烷、四氢呋喃、氯仿、乙腈，优选甲苯。采用上述第一有机溶剂能够为氧化反应提供良好的反应氛围，保证反应顺利进行。
中间体B与所述第一氧化剂的摩尔比为1:0.5-3，优选为1:0.8-2，如在不同实施方式中可以为1﹕1、1﹕1.5、1﹕2、1﹕2.5、1﹕3等等。中间体B和硅藻土的摩尔比为1﹕5-15。如在不同实施方式中可以为1﹕6、1﹕8、1﹕10、1﹕12、1﹕15等等。优选的，中间体B和第一有机溶剂的质量比为1﹕5-20。可在该范围内进行调整。采用上述比例能够保证反应过程顺利进行，保证中间体C的产率。
进一步地，反应结束后对氧化反应得到的反应液进行提纯，形成中间体C，提纯方法是：反应结束后，将氧化反应的反应液过滤，收集滤液，浓缩得到残余样品，用甲醇：水＝5:1进行重结晶，过滤得到中间体C。
S3、制备中间体D；
将上述中间体C与4-二甲氨基吡啶、三乙胺、酸酐进行反应1-15h后调节反应液pH至6-7形成中间体D，也就是第二次酯化反应，第二次酯化反应为7位进行酯化反应。具体地，化合物C溶解于第二有机溶剂中，加入4-二甲氨基吡啶、三乙胺、酸酐，室温反应1-15h，而后将反应后的反应液加入水中，调节pH为6-7，分液收集有机相，得到中间体D。第二有机溶剂为乙酸乙酯，酸酐为乙酸酐。
进一步地，调节pH采用的是浓度为1-3M的盐酸进行调节。
中间体C、所述4-二甲氨基吡啶、所述三乙胺、所述酸酐的摩尔比为1﹕0.01-0.5﹕1.5-3﹕0.8-4。且中间体C与第二有机溶剂的质量比为1:10-15。采用上述比例能够保证第二酯化反应顺利进行，保证中间体D产率。
进一步地，将第二次酯化反应得到的有机相进行纯化，形成中间体D，提纯的方法是：用饱和碳酸氢钠溶液和饱和食盐水溶液洗涤有机相，干燥，除去溶剂，得到中间体D。
S4、制备中间体E；
中间体D与第二氧化剂在20-90℃条件下反应3-72h，也就是脱氢反应，该脱氢反应是1,2和4，5位同时脱氢氧化形成烯烃的反应。具体地，中间体D溶解于第三有机溶剂中，加入第二氧化剂和三氟乙酸，氮气保护下加热反应，得到中间体E。
第三有机溶剂优选二甲基亚砜。第二氧化剂为碘试剂，优选为高价碘试剂，更优选为2-碘酰基苯甲酸。采用上述第二氧化剂和第三有机溶剂能够保证中间体E的生产效率。
中间体D和所述第二氧化剂的摩尔比为1:2-5。中间体D和三氟乙酸的摩尔比为1：0.1-0.5且中间体D和第三有机溶剂的质量比为1﹕10-35。
进一步地，反应后对脱氢反应的反应液进行提纯，提纯的方法是：将反应后的反应液加入水中，用乙酸乙酯萃取，分液收集有机相，用水和饱和食盐水溶液洗涤有机相，干燥，除去溶剂，柱层析分离，得到中间体E。进一步优选的，柱层析分离的洗脱液为乙酸乙酯和石油醚，乙酸乙酯和石油醚的体积比为1﹕10-20。采用上述提纯方式能够进一步保证提纯效果，继而保证后续制备得到的鹅去氧胆酸的纯度和产率。
经过上述操作后得到中间体E，而后中间体E依次经过催化加氢还原反应、3位还原反应以及水解反应得到所述鹅去氧胆酸。
S5、制备中间体F；
催化加氢还原反应是1,2和4，5位同时加氢还原的反应，具体地，将所述中间体E与催化剂、三乙胺混合后在20-60℃的条件下进行加氢反应0.5-4h后形成中间体F，具体地，中间体E、催化剂、醇类溶剂和三乙胺进行反应，反应的压力为0.8-1.2MPa。催化剂为钯碳，优选为5％的钯碳，醇类溶剂为甲醇。
优选，中间体E与所述催化剂和三乙胺的摩尔比为1:2-10：0.001-0.5；中间体E与醇类溶剂的质量比为1:10-20。采用上述比例能够保证催化加氢还原反应顺利进行，保证中间体F的产率。
而后将催化加氢还原反应得到的反应液进行提纯，提纯的方法是反应结束后，旋干醇类溶剂，用乙酸乙酯溶解，用盐酸和饱和食盐水洗涤，除去溶剂，得到中间体F。其中，乙酸乙酯的加入质量优选为中间体F的20-30倍。洗涤时，采用相近体积的浓度为1-3M盐酸、饱和食盐水洗涤，优选相同体积。通过上述提纯方法能够保证制备得到的中间体F的产率和从纯度。
S6、制备中间体G；
3位还原反应是将所述中间体F与还原剂进行反应。3位还原反应的意思是位于3位的羰基进行催化加氢反应得到羟基。
具体地，将中间体F溶解于第四有机溶剂中，于0-10℃加入还原剂，室温反应，得到中间体G。还原剂为氢化物，更优选为硼氢化钠。第四有机溶剂优选无水甲醇。其中，反应时间通过薄层色谱法TLC监控，展开剂为体积比为1﹕6-1﹕2的乙酸乙酯和石油醚。采用上述方式进行反应，能够保证反应顺利进行，保证制备得到的中间体G的产率。
中间体F与还原剂的摩尔比为1:0.5-10，中间体F与第四有机溶剂的质量比为1﹕30-50，采用上述比例能够进一步保证中间体G的制备效果。
进一步地，反应后将3位还原反应得到的反应液进行提纯，提纯方法是：向反应后的反应液中加入水、乙酸乙酯，充分搅拌，静置分层，收集有机相，依次用饱和碳酸氢钠和饱和食盐水洗涤，干燥，除去溶剂，得到中间体F。提纯时，加入水和乙酸乙酯的质量分别优选为反应液质量的5-10倍和10-30倍。
S7、制备鹅去氧胆酸；
水解反应是在碱性条件下，中间体G水解得到鹅去氧胆酸，也就是3α，7α-二羟基-5β-胆烷酸。具体地，中间体G、醇类溶剂、水、碱金属氢氧化物混合，回流反应，浓缩除去醇类溶剂，加入水、盐酸，调节pH为3-4，析出沉淀，过滤收集沉淀，得到鹅去氧胆酸。进一步优选的，于60-80℃反应4-8h。醇类溶剂为甲醇，碱金属氢氧化物包括氢氧化钠和氢氧化钾，优选氢氧化钠。采用上述制备过程，保证生产效率和纯度。
进一步地，中间体G、醇类溶剂和水的质量比为1﹕10-20﹕5-10。中间体G与碱金属氢氧化物的摩尔比为1﹕2-6。采用上述比例能够保证反应充分进行，减少副产物的产生。
其中，加入水的量优选与反应液同体积。加入的盐酸的浓度为1-3M。
本申请还提供一种鹅去氧胆酸，其通过上述鹅去氧胆酸的制备方法制备得到。
综上，本申请各步骤的反应条件相对温和，且工艺简单，经过7步反应得到产物，易于控制，无需复杂的设备即可得到目标化合物。
以下结合实施例对本申请的特征和性能作进一步的详细描述。
实施例1
本实施例提供一种鹅去氧胆酸的制备方法，包括以下步骤：
S1、制备中间体B；
向配有干燥回流冷凝管的反应容器中加入别鹅去氧胆酸(1.0g，2.6mmol)、15mL无水甲醇、浓硫酸100μL，加入完毕后将反应温度升至67℃，搅拌反应4h。反应结束后，旋转蒸发除去溶剂甲醇，加20mL乙酸乙酯溶解残留物，依次用10mL饱和NaHCO 3溶液和水洗涤。将有机相用无水MgSO 4干燥除水，过滤除去无水MgSO 4，收集有机相，减压蒸馏除去溶剂，得到1.0g白色固体，为中间体B，收率：99％。
中间体B的结构表征数据如下：
1H NMR(600MHz,CD 3OD)δ＝4.03-3.90(m,1H),3.77(d,J＝2.1Hz,1H),3.64(s,1H),0.93(dd,J＝13.1,8.9Hz,1H),0.80(s,1H),0.68(s,1H)。
13C NMR(151MHz,CD 3OD)δ＝175.06,67.36,65.81,55.85,50.61,50.34,45.63,42.30,39.52,39.51,36.35,35.81,35.34, 35.10,31.90,31.18,30.85,30.43,28.13,27.74,23.11,20.31,17.36,10.86,9.24。
HRMS:Calcd for C 25H 42O 4[M+Na] +429.2983，Found 429.2971。
S2、制备中间体C；
将中间体B(0.5g，1.23mmol)溶于甲苯(10mL)中，加入硅藻土(0.74g，12.23mmol)，搅拌均匀，避光的情况下加入碳酸银(0.68g，2.46mmol)，升温至回流，搅拌反应24h，过滤除去沉淀，浓缩甲苯，加入3mL甲醇和水(甲醇：水＝5:1)进行重结晶，抽滤，得到中间体C，收率为83％。
中间体C的结构表征数据如下：
1H NMR(600MHz,CD 3OD)δ＝3.79(d,J＝2.6Hz,1H),3.64(s,3H),1.05(s,3H),0.94(d,J＝6.6Hz,3H),0.72(s,3H)。
13C NMR(151MHz,CD 3OD)δ＝213.32,175.02,66.81,55.88,50.62,50.24,45.08,43.69,42.30,39.46,39.30,38.23),37.56,36.53,35.46,35.33,30.82,30.43,27.76,23.13,20.84,17.35,10.86,9.30。
HRMS:Calcd for C 25H 40O 4[M+Na] +427.2827，Found 427.2822。
S3、制备中间体D；
在反应容器中加入中间体C(0.2g，0.49mmol)和乙酸乙酯(3mL)搅拌溶解；然后依次加入DMAP(6mg，0.049mmol)、三乙胺(170μL，1.23mmol)，加入乙酸酐(151μL，1.61mmol)，室温搅拌反应10.5h。加入0.5N盐酸调节pH至6-7，分液收集有机相，有机相依次用水(3×5mL)和饱和食盐水(3×5mL)洗涤，无水MgSO 4干燥除 水，抽滤除去无水MgSO 4，浓缩除去溶剂，得白色固体0.22g，为中间体D，收率为99.5％。
中间体D的结构表征数据如下：
1H NMR(600MHz,CD 3OD)δ＝4.94-4.90(m,1H),3.64(s,1H),2.03(s,1H),1.07(d,J＝7.5Hz,1H),0.94(t,J＝4.9Hz,1H),0.73(d,J＝6.8Hz,1H)。
13C NMR(151MHz,CD 3OD)δ＝212.70,174.96,171.06,70.99(s),55.79,50.61,50.31,46.53,43.32,42.46,40.09,39.34,38.01,38.00,37.46,35.32,35.26,33.01,30.76,30.40,27.58,23.22,20.96,19.73,17.31,10.76,9.33。
HRMS:Calcd for C 27H 42O 5[M+Na] +469.2932，Found 469.2920。
S4、制备中间体E；
将中间体D(1.0g，2.24mmol)和2-碘酰基苯甲酸(2.5g，8.96mmol)加入到反应容器中，在氮气的氛围下加入DMSO(30mL)搅拌溶解，加入三氟乙酸(0.1mL，0.67mmol)，升温至65℃，薄层色谱法TLC(V乙酸乙酯﹕V石油醚＝1﹕3)监控，反应结束后，加入水稀释，用乙酸乙酯(3×20mL)萃取三次，得到的有机相依次用水(3×20mL)和饱和食盐水洗涤(3×20mL)洗涤，无水MgSO 4干燥除水，过滤除去无水MgSO 4，浓缩得粗品，粗品经柱层析分离，洗脱液为体积比为1﹕10的乙酸乙酯和石油醚，得黄色固体0.5g，为中间体E，收率50％。
中间体E的结构表征数据如下：
1H NMR(600MHz,DMSO-d 6)δ＝7.21(d,J＝10.1Hz,1H),6.13(dd,J＝10.1,1.5Hz,1H),5.94(s,1H),4.96(d,J＝2.3Hz,1H), 3.57(s,1H),1.94(s,1H),1.22(s,1H),0.87(d,J＝6.5Hz,1H),0.71(s,1H)。
13C NMR(151MHz,DMSO-d 6)δ＝185.00,174.15,170.25,165.36,156.47,127.35,126.09,72.31,55.52,51.63,50.23,45.49,43.46,42.78,39.03,37.69,37.15,35.13,30.96,30.79,27.83,23.61,22.46,21.24,18.70,18.50,12.09。
HRMS:Calcd for C 27H 40O 5[M+Na] +465.2619，Found 465.2624。
S5、制备中间体F；
将中间体E(0.1g，0.22mmol)溶于甲醇(2mL)中，加入钯碳(0.2g,1.88mmol)，滴加两滴三乙胺，将反应体系置于1MPa的室温下反应1h，旋干甲醇，加入乙酸乙酯(2mL)，依次用1M盐酸(3×2mL)和饱和食盐水(3×2mL)洗涤，无水MgSO 4干燥除水，过滤除去无水MgSO 4，浓缩得中间体F，收率99％。
中间体F的结构表征数据如下：
1H NMR(600MHz,CD 3OD)δ＝4.98-4.91(m,1H),3.64(s,1H),2.02(s,1H),1.07(s,1H),0.96(d,J＝6.6Hz,1H),0.74(s,1H)。
13C NMR(151MHz,CD 3OD)δ＝213.86,174.97,170.74,71.33,55.77,50.62,50.25,44.32,42.76,42.52,39.36,37.78,36.33,36.29,35.26,34.75,34.70,30.77,30.62,30.40,27.64,23.19,20.78,20.75,19.91,17.34,10.73。
HRMS:Calcd for C 27H 44O 5[M+Na] +469.2932，Found 469.2934。
S6、制备中间体G；
将中间体F(34mg，0.076mmol)溶于甲醇(2mL)中，降温至0℃，分批加入硼氢化钠(14mg，0.368mmol)，加毕，反应体系缓慢升至室温下搅拌反应，薄层色谱法TLC(V乙酸乙酯﹕V石油醚＝1﹕2)监控。反应结束后，加入蒸馏水(5mL)，加入乙酸乙酯(20mL)，充分搅拌，静置分层，分出有机相，水相用乙酸乙酯(2×5mL)萃取，合并有机相，依次用饱和碳酸氢钠(3×15mL)和饱和食盐水(3×15mL)洗涤，无水MgSO 4干燥，过滤除去无水MgSO 4，浓缩得白色固体30mg，为中间体G，收率88％。
中间体G的结构表征数据如下：
1H NMR(600MHz,CD 3OD)δ＝4.85(d,J＝2.8Hz,1H),3.64(s,1H),3.41(tt,J＝11.1,4.3Hz,1H),2.04(d,J＝3.7Hz,1H),0.95(s,1H),0.94(d,J＝6.6Hz,1H),0.70(s,1H)。
13C NMR(151MHz,CD 3OD)δ＝174.99,171.13,71.64,70.96,55.80,50.62,50.39,42.45,41.19,39.54,38.44,37.83,35.27,34.90,34.47,34.13,31.08,30.78,30.41,29.97,27.66,23.19,21.87,20.39,20.06,17.34,10.72。
HRMS:Calcd for C 27H 44O 5[M+Na] +471.3089，Found 471.3076。
S7、制备鹅去氧胆酸；
室温下，将中间体G(20mg，0.044mmol)加入到甲醇(0.5mL)和水(0.1mL)的混合溶液中，加入氢氧化钠(12mg，0.3mmol)，回流搅拌6h，浓缩除去甲醇，加入1mL水稀释，然后用1N盐酸调pH至3-4，析出白色沉淀，过滤，干燥，得白色固体16mg，为鹅去氧胆酸，收率90.9％，纯度98.7％。
鹅去氧胆酸的结构表征数据如下：
1H NMR(600MHz,CD 3OD)δ＝3.79(d,J＝2.4Hz,1H),3.41–3.32(m,1H),0.96(d,J＝6.5Hz,1H),0.92(d,J＝10.1Hz,1H),0.69(s,1H)。
13C NMR(151MHz,CD 3OD)δ＝176.80,71.46,67.66,55.93,50.13,42.28,41.78,39.65,39.37,39.07,35.37,35.17,34.83,34.49,32.65,30.95,30.59,29.96,27.84,23.23,22.01,20.39,17.41,10.78。
HRMS:Calcd for C 24H 40O 4[M+H] +393.2927，Found 393.2097。
本实施例还提供一种鹅去氧胆酸，其通过上述制备方法制备得到。总收率为32，纯度为98.7％。
实施例2-实施例10
实施例2-实施例10提供的鹅去氧胆酸的制备方法与实施例1提供的鹅去氧胆酸的制备方法操作基本相似，区别在于采用的原料和各个步骤的具体条件不同。
实施例2：
S1、制备中间体B：醇为异丙醇，反应温度为85℃，反应时间为6小时，催化剂为浓硫酸，3α，7α-二羟基-5α-胆烷酸、异丙醇和浓硫酸的摩尔比为1:5：0.5，提纯是乙酸乙酯的用量为3α，7α-二羟基-5α-胆烷酸的10倍。收率为99％。
表征数据为： 1H NMR(600MHz,CDCl 3)δ＝5.03–4.96(m,1H),4.06–4.03(m,1H),3.82(d,J＝2.4Hz,1H),1.23(s,1H),1.22(s,1H),0.91(t,J＝5.9Hz,1H),0.77(s,1H),0.65(s,1H)。
13C NMR(151MHz,CDCl 3)δ＝173.92,68.05,67.37,66.44,55.80,50.64,45.87,42.65,39.52,39.49,36.24,36.16,35.51, 35.35,31.98,31.63,31.51,31.00,28.83,28.09,23.57,21.88,20.55,18.27,11.86,10.15。
HRMS:Calcd for C 27H 46O 4[M+Na] +434.3396，Found 434.3300。
S2、制备中间体C：反应时间为36小时，第一氧化剂为过氧苯甲酸，第一有机溶剂为四氢呋喃，中间体B与所述第一氧化剂的摩尔比为1：0.5，中间体B和硅藻土的摩尔比为1﹕5。
S3、制备中间体D：反应时间为15h，pH为6.2-6.5，调节pH的颜色的浓度为1M,中间体C、所述4-二甲氨基吡啶、所述三乙胺、所述乙酸酐的摩尔比为1﹕0.1﹕1.5﹕2。
S4、制备中间体E：反应温度90℃，反应时间为48h，中间体D和2-碘酰基苯甲酸的摩尔比为1:2，中间体D和三氟乙酸的摩尔比为1：0.5且中间体D和第三有机溶剂的质量比为1﹕10。提纯时乙酸乙酯和石油醚的体积比为1﹕20。
S5、制备中间体F；反应温度60℃，反应时间为0.5h，反应压力0.8MPa，中间体E与5％的钯碳和三乙胺的摩尔比为1:2：0.001；中间体E与甲醇的质量比为1:10。
S6、制备中间体G；加入硼氢化钠的温度为10℃，展开剂为体积比为1﹕5的乙酸乙酯和石油醚，中间体F与硼氢化钠的摩尔比为1:10，中间体F与无水甲醇的质量比为1﹕30。
S7、制备鹅去氧胆酸：调节pH为3，反应温度为80℃，反应时间为4h，中间体G、甲醇和水的质量比为1﹕10﹕5。中间体G与氢氧化钾的摩尔比为1:2，加入的盐酸的浓度为3M。
总产率为30％，纯度为97％。
实施例3
S1、制备中间体B：醇为乙醇，反应温度为50℃，反应时间为3小时，催化剂为浓盐酸，3α，7α-二羟基-5α-胆烷酸、乙醇和浓盐酸的摩尔比为1:15：1，提纯是乙酸乙酯的用量为3α，7α-二羟基-5α-胆烷酸的20倍。
S2、制备中间体C：反应时间为15小时，第一氧化剂为三氧化铬，第一有机溶剂为二氯甲烷，中间体B与所述第一氧化剂的摩尔比为1：1.5，中间体B和硅藻土的摩尔比为1﹕9。
S3、制备中间体D：反应时间为14h，pH为6.2-6.6，调节pH的颜色的浓度为3M,中间体C、所述4-二甲氨基吡啶、所述三乙胺、所述乙酸酐的摩尔比为1﹕0.4﹕2.5﹕2.5。
S4、制备中间体E：反应温度75℃，反应时间为34h，中间体D和2-碘酰基苯甲酸的摩尔比为1:4.5，中间体D和三氟乙酸的摩尔比为1：0.4且中间体D和第三有机溶剂的质量比为1﹕13。提纯时乙酸乙酯和石油醚的体积比为1﹕12。
S5、制备中间体F；反应温度40℃，反应时间为2h，反应压力0.9MPa，中间体E与5％的钯碳和三乙胺的摩尔比为1:5：0.005；中间体E与甲醇的质量比为1:15。
S6、制备中间体G；加入硼氢化钠的温度为7℃，展开剂为体积比为1﹕3的乙酸乙酯和石油醚，中间体F与硼氢化钠的摩尔比为1:9，中间体F与无水甲醇的质量比为1﹕35。
S7、制备鹅去氧胆酸：调节pH为3.5，反应温度为75℃，反应时间为6h，中间体G、甲醇和水的质量比为1﹕18﹕6。中间体G与氢氧化钾的摩尔比为1:5，加入的盐酸的浓度为1.5M。
总产率为28％，纯度为98％。
实施例4
S1、制备中间体B：醇为丙烯醇，反应温度为90℃，反应时间为4小时，催化剂为甲磺酸，3α，7α-二羟基-5α-胆烷酸、丙烯醇和甲磺酸的摩尔比为1:10：0.7，提纯是乙酸乙酯的用量为3α，7α-二羟基-5α-胆烷酸的15倍。
S2、制备中间体C：反应时间为30小时，第一氧化剂为N-溴代丁二酰亚胺，第一有机溶剂为乙腈，中间体B与所述第一氧化剂的摩尔比为1：0.8，中间体B和硅藻土的摩尔比为1﹕12。
S3、制备中间体D：反应时间为13h，pH为6.1-6.8，调节pH的颜色的浓度为2M,中间体C、所述4-二甲氨基吡啶、所述三乙胺、所述乙酸酐的摩尔比为1﹕0.2﹕2﹕3。
S4、制备中间体E：反应温度50℃，反应时间为36h，中间体D和2-碘酰基苯甲酸的摩尔比为1:3，中间体D和三氟乙酸的摩尔比为1：0.2且中间体D和第三有机溶剂的质量比为1﹕11。提纯时乙酸乙酯和石油醚的体积比为1﹕15。
S5、制备中间体F；反应温度20℃，反应时间为4h，反应压力1.2MPa，中间体E与5％的钯碳和三乙胺的摩尔比为1:10：0.01；中间体E与甲醇的质量比为1:20。
S6、制备中间体G；加入硼氢化钠的温度为2℃，展开剂为体积比为1﹕6的乙酸乙酯和石油醚，中间体F与硼氢化钠的摩尔比为1:5，中间体F与无水甲醇的质量比为1﹕40。
S7、制备鹅去氧胆酸：调节pH为3.2，反应温度为70℃，反应时间为5h，中间体G、甲醇和水的质量比为1﹕15﹕8。中间体G与氢氧化钾的摩尔比为1:4，加入的盐酸的浓度为2M。
总产率为28％，纯度为99％。
实施例5
S1、制备中间体B：醇为苄醇，反应温度为70℃，反应时间为5小时，催化剂为苯磺酸，3α，7α-二羟基-5α-胆烷酸、苄醇和苯磺酸的摩尔比为1:7：0.55，提纯是乙酸乙酯的用量为3α，7α-二羟基-5α-胆烷酸的17倍。
S2、制备中间体C：反应时间为2小时，第一氧化剂为二氧化锰，第一有机溶剂为氯仿，中间体B与所述第一氧化剂的摩尔比为1：3，中间体B和硅藻土的摩尔比为1﹕15。
S3、制备中间体D：反应时间为10h，pH为6.7-6.9，调节pH的颜色的浓度为1.5M,中间体C、所述4-二甲氨基吡啶、所述三乙胺、所述乙酸酐的摩尔比为1﹕0.5﹕3﹕3.5。
S4、制备中间体E：反应温度20℃，反应时间为12h，中间体D和2-碘酰基苯甲酸的摩尔比为1:5，中间体D和三氟乙酸的摩尔比为1：0.1且中间体D和第三有机溶剂的质量比为1﹕15。提纯时乙酸乙酯和石油醚的体积比为1﹕10。
S5、制备中间体F；反应温度50℃，反应时间为3h，反应压力1.1MPa，中间体E与5％的钯碳和三乙胺的摩尔比为1:7：0.008；中间体E与甲醇的质量比为1:17。
S6、制备中间体G；加入硼氢化钠的温度为0℃，展开剂为体积比为1﹕2的乙酸乙酯和石油醚，中间体F与硼氢化钠的摩尔比为1:2，中间体F与无水甲醇的质量比为1﹕50。
S7、制备鹅去氧胆酸：调节pH为4，反应温度为60℃，反应时间为8h，中间体G、甲醇和水的质量比为1﹕20﹕10。中间体G与氢氧化钾的摩尔比为1:6，加入的盐酸的浓度为1M。
总产率为26％，纯度为98％。
实施例6
本实施例与实施例1的制备方法的区别在于中间体C的制备：将中间体B(0.5g，1.23mmol)溶于二氯甲烷(10mL)中，加入活化好的二氧化锰(1.6g，12.3mmol)，室温下反应，薄层色谱法TLC(V 乙酸乙酯﹕V 石油醚＝1﹕1)监控，到原料不再减少停止反应，抽滤，浓缩得粗品，粗品经柱层析分离，洗脱液为体积比为1﹕1的乙酸乙酯和石油醚，得白色固体0.3g，为中间体C，收率60％。总产率为24％，纯度为98％。
实施例7
本实施例与实施例1的制备方法的区别在于，制备中间体D时，乙酸酐的用量为0.98mmol。总产率为30％，纯度为99％。
实施例8
本实施例与实施例1的制备方法的区别在于，制备中间体D时，乙酸酐的用量为1.72mmol。总产率为31％，纯度为98％。
实施例9
本实施例与实施例1的制备方法的区别在于，制备中间体E时，2-碘酰基苯甲酸的用量为4.48mmol。总产率为28％，纯度为98％。
实施例10
本实施例与实施例1的制备方法的区别在于，制备中间体G时，硼氢化钠的用量为0.148mmol。总产率为29％，纯度为98％。
实施例11
本实施例与实施例1的制备方法的区别在于，制备中间体G时，硼氢化钠的用量为0.74mmol。总产率为31％，纯度为98％。
综上所述，本申请的通过采用鸭胆、鹅胆等经过提取后被丢弃的副产物作为反应原料，能够做到废物利用，降低合成成本，且原料来 源广泛，供应充足。并且，经过上述步骤得到的鹅去氧胆酸的产率高，可达32％，适合大量制备。且该制备方法操作简单，重复性强，具有极强的实用性。
以上所述仅为本申请的优选实施方式而已，并不用于限制本申请，对于本领域的技术人员来说，本申请可以有各种更改和变化。凡在本申请的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本申请的保护范围之内。

Claims (10)

  1. 一种鹅去氧胆酸的制备方法，其特征在于，包括以下步骤：
    利用3α，7α-二羟基-5α-胆烷酸为原料经过化学反应后形成中间体E，而后中间体E再经过化学反应形成鹅去氧胆酸；所述中间体E的结构式如下：
    Figure PCTCN2019091965-appb-100001
    其中，R1为烷基、烯基或者芳香基团，R2为酰基。
  2. 根据权利要求1所述的鹅去氧胆酸的制备方法，其特征在于，所述中间体E是将所述3α，7α-二羟基-5α-胆烷酸依次经过第一次酯化反应、氧化反应、第二次酯化反应以及脱氢反应后得到的中间体；
    优选地，第一次酯化反应为3α，7α-二羟基-5α-胆烷酸与醇进行的酯化反应；
    优选地，氧化反应为3位选择性氧化；
    优选地，第二次酯化反应为7位进行酯化反应；
    优选，脱氢反应是1,2和4，5位同时脱氢形成烯烃的反应。
  3. 根据权利要求2所述的鹅去氧胆酸的制备方法，其特征在于，第一次酯化反应是在催化剂的作用下，将所述3α，7α-二羟基-5α-胆烷酸与醇在50-90℃的条件下，反应得到中间体B；
    优选，所述3α，7α-二羟基-5α-胆烷酸、所述醇和所述催化剂的摩尔比为1:2-30:0.1-1；
    优选，所述醇为一元醇，更优选为C1-C10一元醇，进一步优选为甲醇、乙醇、异丙醇、丙烯醇和苄醇，最优选为，甲醇或者异丙醇；
    优选，所述催化剂为酸性物质，更优选为浓盐酸、浓硫酸、甲磺酸、苯磺酸、对甲苯磺酸中的任一种，更优选的，为浓盐酸或者浓硫酸。
  4. 根据权利要求3所述的鹅去氧胆酸的制备方法，其特征在于，氧化反应是将所述中间体B与第一氧化剂反应2-36h形成中间体C；
    优选，所述中间体B与所述第一氧化剂的摩尔比为1:0.5-3，优选为1:0.8-2；
    优选，所述第一氧化剂为过氧化物、金属化合物或者溴代酰亚胺中的任意一种；更优选，所述过氧化物为过氧苯甲酸或者双氧水；所述金属化合物为次氯酸钠、碳酸银、二氧化锰或者三氧化铬中的任意一种；所述溴代酰亚胺为N-溴代丁二酰亚胺；最优选，所述第一氧化剂为碳酸银、次氯酸钠或者二氧化锰。
  5. 根据权利要求4所述的鹅去氧胆酸的制备方法，其特征在于，第二酯化反应是将所述中间体C与4-二甲氨基吡啶、三乙胺、酸酐进行反应1-15h后调节反应液pH至6-7形成中间体D；
    优选，所述中间体C、所述4-二甲氨基吡啶、所述三乙胺、所述酸酐的摩尔比为1﹕0.01-0.5﹕1.5-3﹕0.8-4。
  6. 根据权利要求5所述的鹅去氧胆酸的制备方法，其特征在于，脱氢反应是将所述中间体D与第二氧化剂在20-90℃条件下反应3-72h；
    优选，所述第二氧化剂为碘试剂，优选为高价碘试剂，更优选为2-碘酰基苯甲酸；
    优选，所述中间体D和所述第二氧化剂的摩尔比为1:2-5。
  7. 根据权利要求1所述的鹅去氧胆酸的制备方法，其特征在于，所述中间体E依次经过催化加氢还原反应、3位还原反应以及水解反应得到所述鹅去氧胆酸；
    优选，催化加氢还原反应是1,2和4，5位同时加氢还原的反应。
  8. 根据权利要求7所述的鹅去氧胆酸的制备方法，其特征在于，催化加氢还原反应是将所述中间体E与催化剂、三乙胺混合后在20-60℃的条件下进行加氢反应0.5-4h后形成中间体F；
    优选，所述中间体E与所述催化剂和三乙胺的摩尔比为1:0.02-10：0.001-0.5；
    优选，所述催化剂为钯碳催化剂，优选为5％-10％的钯碳。
  9. 根据权利要求8所述的鹅去氧胆酸的制备方法，其特征在于，3位还原反应是将所述中间体F与还原剂进行反应；
    优选，所述中间体F与还原剂的摩尔比为1:0.5-10；
    优选，所述还原剂为氢化物，更优选为，硼氢化钠。
  10. 一种鹅去氧胆酸，其特征在于，其通过权利要求1所述的鹅去氧胆酸的制备方法制备得到。
PCT/CN2019/091965 2019-03-06 2019-06-20 鹅去氧胆酸及其制备方法 Ceased WO2020177240A1 (zh)

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CN115466300A (zh) * 2022-10-18 2022-12-13 湖南科瑞生物制药股份有限公司 一种胆酸中间体a7及其合成方法
CN115611961A (zh) * 2022-10-18 2023-01-17 湖南科瑞生物制药股份有限公司 胆酸中间体a2及其制备方法
CN115637281A (zh) * 2022-10-31 2023-01-24 湖南科瑞生物制药股份有限公司 制3β,7β-二羟基-5α-H的UDCA异构体方法
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