Chemical-enzymatic preparation of UDCA

A technology for the preparation of ursodeoxycholic acid and enzymatic method, which is applied in the field of chemical-enzymatic preparation of ursodeoxycholic acid, can solve the problems of difficult industrial application, stay in the laboratory research stage, low substrate concentration, etc., and achieve the reaction The effect of mild conditions, cheap raw materials, and easy availability of raw materials

Inactive Publication Date: 2017-05-24
SYNCOZYMES SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] In the enzymatic preparation step of this route, the substrate concentration is as low as 20g/L, w

Method used

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  • Chemical-enzymatic preparation of UDCA
  • Chemical-enzymatic preparation of UDCA
  • Chemical-enzymatic preparation of UDCA

Examples

Experimental program
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Embodiment 1

[0038] Example 1 Synthesis of ursodeoxycholic acid (compound I)

[0039] (1) Preparation of 12-ketodeoxycholic acid (compound III)

[0040] Add 50mM potassium phosphate buffer (10mL, pH=8.0) into a 30mL reaction vessel, add compound II (1.5g), NADP (1.5mg), acetone (1.5mL), stir well and then add 12α-HSDH101 enzyme powder ( 0.15 g) and alcohol dehydrogenase KRED195 (30 mg) were adjusted to 15 mL, and reacted with magnetic stirring at 35°C for 24 hours. After the reaction, heat at 70°C for 2 hours to denature the protein in the reaction solution, remove the protein by filtration, adjust the pH to 3 with 1M hydrochloric acid, extract three times with an equal volume of ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and distill under reduced pressure to obtain 12- Ketodeoxycholic acid (compound III, 1.38 g, yield 92%).

[0041] (2) Preparation of gallstone acid (compound IV)

[0042] Add compound III (1.38g), ethylene glycol (10ml), 85% hydrazine...

Embodiment 2

[0045] The synthesis of embodiment two ursodeoxycholic acid (compound I)

[0046] (1) Preparation of 12-ketodeoxycholic acid (compound III)

[0047]Add 60mM potassium phosphate buffer solution (10mL, pH=7.0) into a 50mL reaction vessel, add compound II (1.5g), NADP (1.5mg), acetone (1.5mL), stir well and then add 12α-HSDH101 enzyme powder ( 150mg) and alcohol dehydrogenase KRED195 (30mg) were adjusted to 30mL, and reacted with magnetic stirring at 40°C for 24 hours. After the reaction, heat at 70°C for 2 hours to denature the protein in the reaction solution, remove the protein by filtration, adjust the pH to 3 with 1M hydrochloric acid, extract three times with an equal volume of ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and distill under reduced pressure to obtain 12- Ketodeoxycholic acid (compound III, 1.43 g, yield 94%).

[0048] (2) Preparation of gallstone acid (compound IV)

[0049] Add compound III (1.43g), ethylene glycol (15ml),...

Embodiment 3

[0052] Example 3 Synthesis of ursodeoxycholic acid (compound I)

[0053] (1) Preparation of 12-ketodeoxycholic acid (compound III)

[0054] Add 40mM potassium phosphate buffer solution (10mL, pH=7.0) into a 100mL reaction vessel, add compound II (1.5g), NADP (1.0mg), acetone (1.5mL), stir well and then add 12α-HSDH101 enzyme powder ( 300mg) and alcohol dehydrogenase KRED195 (50mg) were adjusted to 50mL, and reacted with magnetic stirring at 30°C for 20 hours. After the reaction, heat at 70°C for 2 hours to denature the protein in the reaction solution, remove the protein by filtration, adjust the pH to 3 with 1M hydrochloric acid, extract three times with an equal volume of ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, and distill under reduced pressure to obtain 12- Ketodeoxycholic acid (compound III, 1.34 g, yield 90%).

[0055] (2) Preparation of gallstone acid (compound IV)

[0056] Add compound III (1.34g), ethylene glycol (15ml), 85% hy...

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Abstract

The invention discloses a chemical-enzymatic preparation of UDCA. The chemical-enzymatic preparation of UDCA is characterized by comprising the following synthesis procedures: a) oxidizing UDCA under the action of 12alpha-HSDH to obtain 12-ketodeoxycholic acid, b) carrying out a decarbonylation reduction reaction on 12-ketodeoxycholic acid to obtain lithocholic acid, and c) carrying out hydroxylation on the lithocholic acid under the action of 7alpha-LCAH to obtain UDCA. Compared with the prior art, the chemical-enzymatic preparation of UDCA disclosed by the invention is low-cost and readily available in raw materials, mild in reaction conditions, less in steps and simple in post-treatment, and the substrate concentration reaches up to 100g /L, the total yield is 73%, the product purity is more than 99%, and the industrial large-scale production has very good application prospects.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a chemical-enzymatic method for preparing ursodeoxycholic acid. Background technique [0002] Ursodeoxycholic acid (UDCA) was the first pure crystalline product isolated from bear bile by Shoda in Japan in 1927. In 1957, UDCA was successfully synthesized and used clinically as a choleretic. In 1973, Japan discovered that UDCA has the effect of dissolving gallstones, and has the advantages of low toxicity and small dose, which has attracted the attention of countries all over the world. Ursodeoxycholic acid is a non-toxic hydrophilic cholic acid, which can competitively inhibit the absorption of toxic endogenous cholic acid in the ileum, by activating the signaling network composed of calcium ions and protein kinase C, and by activating the splitting active protein Kinase to enhance the secretory ability of cholestatic liver cells, reduce the concentration of endogenous hyd...

Claims

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Application Information

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IPC IPC(8): C12P33/06C12P33/02
CPCC12P33/02C12P33/06
Inventor 竺伟高新星
Owner SYNCOZYMES SHANGHAI
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