CN114276401A - Method for synthesizing 24-norursodeoxycholic acid - Google Patents
Method for synthesizing 24-norursodeoxycholic acid Download PDFInfo
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- CN114276401A CN114276401A CN202111617475.1A CN202111617475A CN114276401A CN 114276401 A CN114276401 A CN 114276401A CN 202111617475 A CN202111617475 A CN 202111617475A CN 114276401 A CN114276401 A CN 114276401A
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- QYYDXDSPYPOWRO-JHMCBHKWSA-N (3r)-3-[(3r,5s,7s,8r,9s,10s,13r,14s,17r)-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]butanoic acid Chemical compound C([C@H]1C[C@@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CC(O)=O)C)[C@@]2(C)CC1 QYYDXDSPYPOWRO-JHMCBHKWSA-N 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 23
- 230000002194 synthesizing effect Effects 0.000 title claims abstract description 16
- 239000002253 acid Substances 0.000 claims abstract description 17
- 239000007787 solid Substances 0.000 claims description 64
- 150000001875 compounds Chemical class 0.000 claims description 59
- 239000012071 phase Substances 0.000 claims description 59
- 239000002904 solvent Substances 0.000 claims description 58
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 50
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 48
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 45
- 238000006243 chemical reaction Methods 0.000 claims description 42
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 33
- 102000004190 Enzymes Human genes 0.000 claims description 32
- 108090000790 Enzymes Proteins 0.000 claims description 32
- LVTJOONKWUXEFR-FZRMHRINSA-N protoneodioscin Natural products O(C[C@@H](CC[C@]1(O)[C@H](C)[C@@H]2[C@]3(C)[C@H]([C@H]4[C@@H]([C@]5(C)C(=CC4)C[C@@H](O[C@@H]4[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@@H](O)[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@H](CO)O4)CC5)CC3)C[C@@H]2O1)C)[C@H]1[C@H](O)[C@H](O)[C@H](O)[C@@H](CO)O1 LVTJOONKWUXEFR-FZRMHRINSA-N 0.000 claims description 32
- 230000001590 oxidative effect Effects 0.000 claims description 31
- 238000001914 filtration Methods 0.000 claims description 24
- 239000012074 organic phase Substances 0.000 claims description 24
- 239000007800 oxidant agent Substances 0.000 claims description 22
- 238000003756 stirring Methods 0.000 claims description 22
- 239000003638 chemical reducing agent Substances 0.000 claims description 18
- 238000001816 cooling Methods 0.000 claims description 18
- 238000001035 drying Methods 0.000 claims description 18
- 238000005406 washing Methods 0.000 claims description 18
- JNGZXGGOCLZBFB-IVCQMTBJSA-N compound E Chemical compound N([C@@H](C)C(=O)N[C@@H]1C(N(C)C2=CC=CC=C2C(C=2C=CC=CC=2)=N1)=O)C(=O)CC1=CC(F)=CC(F)=C1 JNGZXGGOCLZBFB-IVCQMTBJSA-N 0.000 claims description 14
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 claims description 14
- 238000007254 oxidation reaction Methods 0.000 claims description 13
- 238000005292 vacuum distillation Methods 0.000 claims description 13
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- 238000010791 quenching Methods 0.000 claims description 12
- 238000010438 heat treatment Methods 0.000 claims description 11
- 102000003855 L-lactate dehydrogenase Human genes 0.000 claims description 10
- 108700023483 L-lactate dehydrogenases Proteins 0.000 claims description 10
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- 239000000706 filtrate Substances 0.000 claims description 10
- 229910052755 nonmetal Inorganic materials 0.000 claims description 10
- 230000000694 effects Effects 0.000 claims description 8
- -1 nicotinamide adenine dinucleotide phosphate compound Chemical class 0.000 claims description 8
- 150000003839 salts Chemical class 0.000 claims description 8
- 108010032887 7 beta-hydroxysteroid dehydrogenase Proteins 0.000 claims description 7
- 229940126062 Compound A Drugs 0.000 claims description 7
- NLDMNSXOCDLTTB-UHFFFAOYSA-N Heterophylliin A Natural products O1C2COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC2C(OC(=O)C=2C=C(O)C(O)=C(O)C=2)C(O)C1OC(=O)C1=CC(O)=C(O)C(O)=C1 NLDMNSXOCDLTTB-UHFFFAOYSA-N 0.000 claims description 7
- 108010026217 Malate Dehydrogenase Proteins 0.000 claims description 7
- 102000013460 Malate Dehydrogenase Human genes 0.000 claims description 7
- 229940054269 sodium pyruvate Drugs 0.000 claims description 7
- 101710088194 Dehydrogenase Proteins 0.000 claims description 6
- 238000004440 column chromatography Methods 0.000 claims description 6
- 238000006722 reduction reaction Methods 0.000 claims description 6
- 239000001394 sodium malate Substances 0.000 claims description 6
- 150000004706 metal oxides Chemical class 0.000 claims description 5
- 238000000926 separation method Methods 0.000 claims description 5
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000000460 chlorine Substances 0.000 claims description 4
- 229910052801 chlorine Inorganic materials 0.000 claims description 4
- JBTWLSYIZRCDFO-UHFFFAOYSA-N ethyl methyl carbonate Chemical compound CCOC(=O)OC JBTWLSYIZRCDFO-UHFFFAOYSA-N 0.000 claims description 4
- JGJLWPGRMCADHB-UHFFFAOYSA-N hypobromite Chemical compound Br[O-] JGJLWPGRMCADHB-UHFFFAOYSA-N 0.000 claims description 4
- WQYVRQLZKVEZGA-UHFFFAOYSA-N hypochlorite Chemical compound Cl[O-] WQYVRQLZKVEZGA-UHFFFAOYSA-N 0.000 claims description 4
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 claims description 4
- 150000002576 ketones Chemical class 0.000 claims description 4
- 229910044991 metal oxide Inorganic materials 0.000 claims description 4
- 150000002978 peroxides Chemical class 0.000 claims description 4
- 239000013078 crystal Substances 0.000 claims description 3
- 230000000171 quenching effect Effects 0.000 claims description 3
- 239000000126 substance Substances 0.000 claims description 3
- 238000001308 synthesis method Methods 0.000 claims description 3
- 238000002425 crystallisation Methods 0.000 claims description 2
- 230000008025 crystallization Effects 0.000 claims description 2
- 229910052751 metal Inorganic materials 0.000 claims description 2
- 239000002184 metal Substances 0.000 claims description 2
- 229910052987 metal hydride Inorganic materials 0.000 claims description 2
- 150000004681 metal hydrides Chemical class 0.000 claims description 2
- XJLXINKUBYWONI-NNYOXOHSSA-N NADP zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-NNYOXOHSSA-N 0.000 claims 3
- 102100039358 3-hydroxyacyl-CoA dehydrogenase type-2 Human genes 0.000 claims 1
- 108010014831 7-alpha-hydroxysteroid dehydrogenase Proteins 0.000 claims 1
- 150000001348 alkyl chlorides Chemical class 0.000 claims 1
- WPUMTJGUQUYPIV-JIZZDEOASA-L disodium (S)-malate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](O)CC([O-])=O WPUMTJGUQUYPIV-JIZZDEOASA-L 0.000 claims 1
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 abstract description 9
- 239000004380 Cholic acid Substances 0.000 abstract description 9
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 abstract description 9
- 229960002471 cholic acid Drugs 0.000 abstract description 9
- 235000019416 cholic acid Nutrition 0.000 abstract description 9
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 abstract description 9
- 239000002994 raw material Substances 0.000 abstract description 7
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 238000006479 redox reaction Methods 0.000 abstract description 2
- MSXVEPNJUHWQHW-UHFFFAOYSA-N 2-methylbutan-2-ol Chemical compound CCC(C)(C)O MSXVEPNJUHWQHW-UHFFFAOYSA-N 0.000 description 23
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 21
- RUDATBOHQWOJDD-UHFFFAOYSA-N (3beta,5beta,7alpha)-3,7-Dihydroxycholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 RUDATBOHQWOJDD-UHFFFAOYSA-N 0.000 description 20
- 229960001091 chenodeoxycholic acid Drugs 0.000 description 16
- 238000002360 preparation method Methods 0.000 description 16
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 15
- 238000005070 sampling Methods 0.000 description 15
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 12
- 239000005708 Sodium hypochlorite Substances 0.000 description 11
- 239000008346 aqueous phase Substances 0.000 description 11
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 11
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 description 9
- XJLXINKUBYWONI-NNYOXOHSSA-O NADP(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-NNYOXOHSSA-O 0.000 description 9
- FUZZWVXGSFPDMH-UHFFFAOYSA-M hexanoate Chemical compound CCCCCC([O-])=O FUZZWVXGSFPDMH-UHFFFAOYSA-M 0.000 description 9
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 6
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- UKLNMMHNWFDKNT-UHFFFAOYSA-M sodium chlorite Chemical compound [Na+].[O-]Cl=O UKLNMMHNWFDKNT-UHFFFAOYSA-M 0.000 description 6
- 229960002218 sodium chlorite Drugs 0.000 description 6
- JVBXVOWTABLYPX-UHFFFAOYSA-L sodium dithionite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])=O JVBXVOWTABLYPX-UHFFFAOYSA-L 0.000 description 6
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 5
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 229910001220 stainless steel Inorganic materials 0.000 description 5
- 239000010935 stainless steel Substances 0.000 description 5
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- NTIZESTWPVYFNL-UHFFFAOYSA-N Methyl isobutyl ketone Chemical compound CC(C)CC(C)=O NTIZESTWPVYFNL-UHFFFAOYSA-N 0.000 description 4
- UIHCLUNTQKBZGK-UHFFFAOYSA-N Methyl isobutyl ketone Natural products CCC(C)C(C)=O UIHCLUNTQKBZGK-UHFFFAOYSA-N 0.000 description 4
- KFSLWBXXFJQRDL-UHFFFAOYSA-N Peracetic acid Chemical compound CC(=O)OO KFSLWBXXFJQRDL-UHFFFAOYSA-N 0.000 description 4
- 229910052794 bromium Inorganic materials 0.000 description 4
- BTANRVKWQNVYAZ-UHFFFAOYSA-N butan-2-ol Chemical compound CCC(C)O BTANRVKWQNVYAZ-UHFFFAOYSA-N 0.000 description 4
- 125000003716 cholic acid group Chemical group 0.000 description 4
- 238000006114 decarboxylation reaction Methods 0.000 description 4
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- KMUONIBRACKNSN-UHFFFAOYSA-N potassium dichromate Chemical compound [K+].[K+].[O-][Cr](=O)(=O)O[Cr]([O-])(=O)=O KMUONIBRACKNSN-UHFFFAOYSA-N 0.000 description 4
- 230000035484 reaction time Effects 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 4
- 229960001661 ursodiol Drugs 0.000 description 4
- 239000012065 filter cake Substances 0.000 description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 3
- JMMWKPVZQRWMSS-UHFFFAOYSA-N isopropanol acetate Natural products CC(C)OC(C)=O JMMWKPVZQRWMSS-UHFFFAOYSA-N 0.000 description 3
- 229940011051 isopropyl acetate Drugs 0.000 description 3
- GWYFCOCPABKNJV-UHFFFAOYSA-M isovalerate Chemical compound CC(C)CC([O-])=O GWYFCOCPABKNJV-UHFFFAOYSA-M 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- CRWJEUDFKNYSBX-UHFFFAOYSA-N sodium;hypobromite Chemical compound [Na+].Br[O-] CRWJEUDFKNYSBX-UHFFFAOYSA-N 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- RUDATBOHQWOJDD-UZVSRGJWSA-N ursodeoxycholic acid Chemical compound C([C@H]1C[C@@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 RUDATBOHQWOJDD-UZVSRGJWSA-N 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- AMQJEAYHLZJPGS-UHFFFAOYSA-N N-Pentanol Chemical compound CCCCCO AMQJEAYHLZJPGS-UHFFFAOYSA-N 0.000 description 2
- PCLIMKBDDGJMGD-UHFFFAOYSA-N N-bromosuccinimide Chemical compound BrN1C(=O)CCC1=O PCLIMKBDDGJMGD-UHFFFAOYSA-N 0.000 description 2
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 2
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- KRVSOGSZCMJSLX-UHFFFAOYSA-L chromic acid Substances O[Cr](O)(=O)=O KRVSOGSZCMJSLX-UHFFFAOYSA-L 0.000 description 2
- VTIIJXUACCWYHX-UHFFFAOYSA-L disodium;carboxylatooxy carbonate Chemical compound [Na+].[Na+].[O-]C(=O)OOC([O-])=O VTIIJXUACCWYHX-UHFFFAOYSA-L 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 239000003759 ester based solvent Substances 0.000 description 2
- AWJWCTOOIBYHON-UHFFFAOYSA-N furo[3,4-b]pyrazine-5,7-dione Chemical compound C1=CN=C2C(=O)OC(=O)C2=N1 AWJWCTOOIBYHON-UHFFFAOYSA-N 0.000 description 2
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- BAZAXWOYCMUHIX-UHFFFAOYSA-M sodium perchlorate Chemical compound [Na+].[O-]Cl(=O)(=O)=O BAZAXWOYCMUHIX-UHFFFAOYSA-M 0.000 description 2
- 229910001488 sodium perchlorate Inorganic materials 0.000 description 2
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 2
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- BZSXEZOLBIJVQK-UHFFFAOYSA-N 2-methylsulfonylbenzoic acid Chemical compound CS(=O)(=O)C1=CC=CC=C1C(O)=O BZSXEZOLBIJVQK-UHFFFAOYSA-N 0.000 description 1
- ZKLFRQSZDUSMQE-UHFFFAOYSA-N 5,5-dichloroimidazolidine-2,4-dione Chemical compound ClC1(Cl)NC(=O)NC1=O ZKLFRQSZDUSMQE-UHFFFAOYSA-N 0.000 description 1
- 208000004611 Abdominal Obesity Diseases 0.000 description 1
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- 241001432959 Chernes Species 0.000 description 1
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- 206010022489 Insulin Resistance Diseases 0.000 description 1
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- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
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- 108090000340 Transaminases Proteins 0.000 description 1
- WGLPBDUCMAPZCE-UHFFFAOYSA-N Trioxochromium Chemical compound O=[Cr](=O)=O WGLPBDUCMAPZCE-UHFFFAOYSA-N 0.000 description 1
- GOPYZMJAIPBUGX-UHFFFAOYSA-N [O-2].[O-2].[Mn+4] Chemical class [O-2].[O-2].[Mn+4] GOPYZMJAIPBUGX-UHFFFAOYSA-N 0.000 description 1
- 201000000690 abdominal obesity-metabolic syndrome Diseases 0.000 description 1
- 150000001335 aliphatic alkanes Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910000435 bromine oxide Inorganic materials 0.000 description 1
- 239000012928 buffer substance Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- RUDATBOHQWOJDD-BSWAIDMHSA-N chenodeoxycholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 RUDATBOHQWOJDD-BSWAIDMHSA-N 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
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- 230000029142 excretion Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- CUILPNURFADTPE-UHFFFAOYSA-N hypobromous acid Chemical compound BrO CUILPNURFADTPE-UHFFFAOYSA-N 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- 239000012280 lithium aluminium hydride Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000005895 oxidative decarboxylation reaction Methods 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
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Abstract
The invention discloses a method for synthesizing 24-norursodeoxycholic acid. The method uses the seal cholic acid (3 alpha, 7 alpha, 23 alpha-trihydroxy-5 beta-cholanic acid) as a raw material to obtain the 24-norursodesoxycholic acid through a series of oxidation-reduction reactions, and the method has the advantages of wide raw material source, low price, mild conditions of each step of a synthetic route, high yield and suitability for mass production.
Description
Technical Field
The invention belongs to the field of organic chemical synthesis, and particularly relates to a synthesis method of 24-norursodeoxycholic acid.
Background
The 24-norursodeoxycholic acid is a homolog of ursodeoxycholic acid, has one methylene group less than the side chain of ursodeoxycholic acid, and has the following chemical structural formula:
the 24-norursodeoxycholic acid has relatively weak affinity to taurine and glycine, so that the taurine and glycine are easy to be reabsorbed by bile duct cells to return hepatic sinusoid and hepatic cells, and the excretion of the bile duct cells is promoted.
At present, basic treatment strategies of non-alcoholic fatty liver disease patients mainly comprise life style change, weight loss, abdominal obesity improvement, insulin resistance improvement, drug treatment aiming at metabolic syndrome components and the like; whereas simple basal therapy rarely reverses steatohepatitis. In the annual meeting of liver disease in the united states summoned by 19 to 23 months 10 and 2017, a phase iia randomized double blind control study from the team of austria and germany reported that a large dose of 24-norursodeoxycholic acid can significantly improve serum transaminase, triglyceride and liver imaging indices of nonalcoholic fatty liver disease patients.
However, few methods for synthesizing 24-norursodeoxycholic acid have been reported so far, and almost all methods are synthesized by decarboxylation using ursodeoxycholic acid as a raw material, but the raw material is expensive and the route is long. In contrast, the seal cholic acid is cheap and easily available as a byproduct in chenodeoxycholic acid extraction. And oxidative decarboxylation of seal cholic acid has been reported as early as 1986 (Collection czechlosovak chern. Commun.1986, 51, 1722), and recently also patents have been reported for the synthesis of 24-nor cholestanic acid (CN113135971A, 2021); however, in these syntheses decarboxylation and oxidation of the hydroxyl groups at the 3, 7 positions compete, especially if the hydroxyl group at the 3 position is of the same configuration as the hydroxyl group in the final product, which, if oxidized, would result in the need for an additional step for stereoselective reduction.
Disclosure of Invention
The invention aims to: the 24-norursodeoxycholic acid is obtained by a series of oxidation-reduction reactions by taking seal cholic acid (3 alpha, 7 alpha, 23 alpha-trihydroxy-5 beta-cholanic acid) as a raw material, and has the advantages of wide raw material source, low price, mild conditions of each step of a synthetic route, high yield and suitability for mass production.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for synthesizing 24-norursodeoxycholic acid comprises the following specific steps:
a. dissolving a compound A (seal cholic acid) in a first solvent, and oxidizing by an oxidant 1 in a microchannel reactor to obtain a compound B (24-demethylaldehyde); adding a reducing agent to quench the reaction, standing for layering, discarding the water phase, washing the organic phase with a saturated sodium bicarbonate solution, removing the solvent, and drying to obtain a solid compound B;
b. dissolving a solid compound B (24-demethylaldehyde) in a second solvent, and oxidizing by an oxidant 2 to obtain a compound C (24-demethylchenodeoxycholic acid); adding a reducing agent for quenching reaction, standing for layering, removing a water phase, washing an organic phase by using a saturated sodium chloride solution, removing a solvent to obtain a white solid, dissolving the white solid in ethyl acetate, performing column chromatography separation, and distilling to remove the ethyl acetate to obtain a solid compound C;
c. dissolving the solid compound C (24-demethyl chenodeoxycholic acid) in a third solvent, converting the solid compound C into a compound D (24-demethyl-7-ketolithocholic acid) through oxidation reaction, and purifying and drying the compound D to obtain a solid compound D;
the oxidation reaction is as follows: dissolving the solid compound C in a third solvent, and oxidizing by an oxidant 3 or an enzyme component 1 to obtain a compound D;
d. dissolving the solid compound D (24-demethyl-7-ketolithocholic acid) in a third solvent, and converting the solid compound D (24-demethyl ursodeoxycholic acid) into a compound E through a reduction reaction; purifying and drying the obtained compound E;
the reduction reaction is as follows: dissolving the solid compound D in a third solvent, and reducing by using a reducing agent 1 or an enzyme component 2 to obtain a compound E;
wherein, the reaction process of the synthesis method is shown as a scheme (I):
in step a of the invention, the first solvent is selected from one of acetonitrile, ester solvents (such as ethyl acetate, butyl acetate and isopropyl acetate), ketone solvents (such as acetone and butanone), alcohol solvents (such as tert-butyl alcohol) and chlorinated alkanes (such as dichloromethane and chloroform), and the volume-to-weight ratio of the first solvent to the compound A is 3-50 ml:1 g; the oxidant 1 is one of chlorine, hypochlorous acid, hypochlorite (such as sodium hypochlorite) and hypobromite (such as sodium hypobromite) or any one or more of equivalents (such as N-bromosuccinimide and dichlorohydantoin) obtained through simple reaction, the molar ratio of the oxidant 1 to the compound A is 1-2.5: 1, and the oxidation reaction temperature is-10-50 ℃.
In the step a, the pressure of the microchannel reactor is 0.1-0.3 MPa, and the reactor is made of 1/16-inch stainless steel tubes.
In step B, the second solvent is selected from any one of acetonitrile, ester solvents (such as ethyl acetate, butyl acetate and isopropyl acetate), ketone solvents (such as acetone and butanone) and alcohol solvents (such as tert-butyl alcohol), and the volume weight ratio of the second solvent to the compound B is 3-20 ml:1 g; the oxidant 2 is any one or more of active nonmetal (such as chlorine and bromine), high-valence metal oxide and acid or salt thereof (such as ferric trioxide, activated manganese dioxide, chromic acid, potassium dichromate and potassium permanganate), peroxide (such as hydrogen peroxide, peracetic acid and sodium percarbonate) and acid or salt of non-metal oxide (such as nitric acid, concentrated sulfuric acid, sodium hypochlorite, sodium chlorite and sodium chlorate), the molar ratio of the oxidant 2 to the compound B is 1-3: 1, and the oxidation reaction temperature is 5-40 ℃.
In step C, the third solvent is any one of an ester solvent (such as ethyl acetate, butyl acetate and isopropyl acetate), a ketone solvent (such as acetone and butanone) and an alcohol solvent (such as butanol and pentanol), and the volume-to-weight ratio of the third solvent to the compound C is 5-20 ml:1 g.
In step C of the present invention, the solid compound C is oxidized by an oxidant 3 to obtain a compound D, and the solid compound D is obtained by purification, which comprises the following specific steps:
dissolving the solid compound C in a third solvent, and adding an oxidant 3 to oxidize the compound C into a compound D; adding a reducing agent to quench the reaction, standing for layering, discarding the water phase, washing the organic phase with water, carrying out vacuum distillation to remove water in the organic phase, cooling for crystallization, filtering and drying to obtain a solid compound D.
The oxidant 3 is any one of active nonmetal (such as chlorine and bromine), high valence metal oxide and acid and salt thereof (such as chromic oxide, chromic acid, potassium dichromate and potassium permanganate), peroxide (such as hydrogen peroxide, peracetic acid and sodium percarbonate), acid and salt of nonmetal oxide (such as hypochlorous acid, sodium hypochlorite and sodium chlorite); the molar ratio of the oxidant 3 to the compound C is 1-3: 1; the oxidation reaction temperature is-15 to 20 ℃.
As another embodiment of the present invention, in step C, the solid compound C may also be oxidized by the enzyme component 1 and then purified to obtain the solid compound D, which comprises the following specific steps:
dissolving the solid compound C in a third solvent, adding an enzyme component 1, reacting for 1-12 h at the temperature of 20-40 ℃ and the pH value of the solution of 6.0-8.0, and oxidizing to obtain a compound D; heating the reaction solution to 70-75 ℃, stirring for 2h, cooling to 30 ℃, adjusting the pH value of the solution to 4-4.5 with hydrochloric acid, filtering to remove enzyme, standing and layering the filtrate, discarding the water phase, adding 10% sodium hydroxide solution into the organic phase, adjusting the pH value of the solution to 8.0-8.5, removing the solvent by vacuum distillation, adjusting the pH value of the solution to 3-4 with hydrochloric acid, filtering and drying to obtain a solid compound D.
The enzyme component 1 consists of nicotinamide adenine dinucleotide phosphate, sodium pyruvate, lactate dehydrogenase and 7-alpha hydroxyl steroid dehydrogenase, wherein the content of each component is as follows:
the weight ratio of the nicotinamide adenine dinucleotide phosphate compound C is 2-5: 1000;
the weight ratio of the sodium pyruvate to the compound C is 0.3-0.8: 1;
the activity weight ratio of the lactate dehydrogenase to the compound C is 200-2000U: 1 g;
the activity weight ratio of the alpha-hydroxysteroid dehydrogenase to the compound C is 300-1000U:1 g.
In step D of the invention, the specific operation steps of reducing the solid compound D by the reducing agent 1 and then purifying to obtain the solid compound E are as follows:
dissolving a solid compound D in a third solvent, and adding a reducing agent 1 to reduce the compound D into a crude compound E; and adding hydrochloric acid to adjust the pH value of the solution to 4-4.5, layering, washing an organic phase with water, distilling in vacuum until crystals are separated out, cooling, filtering and drying.
The reducing agent 1 is one of active metal (such as lithium, sodium and potassium), active metal hydride (such as sodium borohydride and lithium aluminum hydride) and nonmetal simple substance (such as hydrogen), the molar ratio of the reducing agent 4 to the compound D is 1-10: 1, and the reduction reaction temperature is 10-105 ℃.
As another embodiment of the present invention, in step D, the solid compound D may also be reduced by the enzyme component 2 and then purified to obtain the solid compound E, which comprises the following specific steps:
dissolving a solid compound D in a third solvent, adding an enzyme component 2, reacting for 1-10 hours at the temperature of 10-40 ℃ and the pH value of the solution of 6.0-8.0, and reducing the compound D into a compound E; heating the reaction solution to 70-75 ℃, stirring for 2h, cooling to 30 ℃, adjusting the pH value of the solution to 4-4.5 with hydrochloric acid, filtering to remove enzyme, standing and layering the filtrate, discarding the water phase, adding 10% sodium hydroxide solution into the organic phase, adjusting the pH value of the solution to 8.0-8.5, removing the solvent by vacuum distillation, adjusting the pH value of the solution to 3-4 with hydrochloric acid, filtering, and drying.
The enzyme component 2 consists of nicotinamide adenine dinucleotide phosphate, L-sodium malate, malate dehydrogenase and 7-beta hydroxysteroid dehydrogenase, wherein the content of each component is as follows:
the weight ratio of the nicotinamide adenine dinucleotide phosphate compound D is 2-5: 1000;
the weight ratio of the L-sodium malate to the compound D is 0.5-1.0: 1;
the activity weight ratio of the malate dehydrogenase to the compound D is 150-1800U: 1 g;
the activity weight ratio of the 7-beta hydroxysteroid dehydrogenase to the compound D is 200-3000U: 1 g.
In the invention, the hydrochloric acid is hydrochloric acid with the concentration of 6M.
In the method for synthesizing 24-norursodeoxycholic acid, the reducing agents for quenching in steps a-c include, but are not limited to, sodium sulfite, sodium bisulfite and sodium hydrosulfite (sodium hydrosulfite), and the reducing agents used in steps a-c can be selected from the same compound or different compounds.
Compared with the prior art, the invention has the beneficial effects that:
the raw materials of the invention are cheap and easily available, and the production cost is reduced;
the invention utilizes the carbon dioxide generated in the decarboxylation reaction as a buffer substance, prevents the overflow of the carbon dioxide by pressurizing the microchannel reactor, and solves the problem of poor selectivity of the oxidation and decarboxylation reaction;
the invention selects hypochlorous acid and hypobromous acid as oxidation reagents, and has the advantages of low price, easy obtaining, safe use, green and environmental protection;
the method has the advantages of mild reaction conditions, simple process, easy amplification, good reaction selectivity and environmental friendliness, and the obtained 24-norursodeoxycholic acid has high yield and good quality and is suitable for mass production.
Detailed Description
The present invention will be further described with reference to the following examples, but is not limited thereto.
Example 1
a. Preparation of 24-demethylaldehyde
Adding 300mL of butyl acetate and 20g of fully crushed seal cholic acid into a 500mL reaction bottle, mixing the obtained micro-turbid liquid with 30mL of sodium hypochlorite with the concentration of 1.5N in a micro mixer, and then feeding the mixture into a micro-channel reactor, wherein the temperature of the micro-channel reactor is 10 ℃, the pressure of the micro-channel reactor is 0.2MPa, the reactor adopts an 1/16-inch stainless steel tube, the volume of the reactor is 1mL, the reaction time is 2-5 min, and the content of seal cholic acid residue is preferably less than 0.5% by sampling after the reaction. Adding a proper amount of sodium hydrosulfite into the reaction solution to quench excessive sodium hypochlorite, standing for layering, and discarding the water phase; adding 400ml of saturated sodium bicarbonate solution into the butyl acetate phase, stirring for 30min, standing for layering, and removing the water phase; repeatedly washing the butyl acetate phase with 400ml of saturated sodium bicarbonate solution once, and discarding the water phase; the butyl acetate phase was distilled under vacuum until the distillate flow off to give about 18g of a white solid (24-demethylaldehyde).
b. Preparation of 24-demethylchenodeoxycholic acid
And (3) completely dissolving 18g of 24-demethyl aldehyde in 360ml of ethyl acetate, slowly adding 90ml of 10% sodium hypochlorite solution in a flowing manner, continuously stirring and reacting for 1h after the sodium hypochlorite is added, and sampling to detect that the substrate residue is 0.331%. Standing for layering, and removing a water phase; adding 400ml of saturated sodium chloride solution into the ethyl acetate phase, stirring for 30min, standing for layering, and removing the water phase; the ethyl acetate phase was washed once more with 400ml of saturated sodium chloride solution and the aqueous phase was discarded. The ethyl acetate phase was vacuum distilled until the distillate flow off to give a white solid. And (3) re-dissolving the white solid in ethyl acetate, performing column chromatography separation, collecting a solvent section containing 24-demethyl chenodeoxycholic acid, and performing vacuum distillation until the flow is cut off to obtain about 10g of 24-demethyl chenodeoxycholic acid.
c. Preparation of 24-demethyl-7-ketolithocholic acid
Dissolving the 24-demethyl chenodeoxycholic acid in 200ml butyl acetate, adding 50mg of Nicotinamide Adenine Dinucleotide Phosphate (NADP), 8g of sodium pyruvate, 20000U of Lactate Dehydrogenase (LDH) and 10000U of 7-alpha hydroxyl steroid dehydrogenase (7-alpha HSDH), reacting for 6h, and sampling to determine that the residue of the 24-demethyl chenodeoxycholic acid is 0.562%. Heating the reaction solution to 70-75 ℃, stirring for 2h to denature the enzyme in the reaction solution, cooling to 30 ℃, adding 5ml of 6M hydrochloric acid, adjusting the pH value of the solution to 4.38, and filtering to remove the enzyme; the filtrate was allowed to stand for separation, the aqueous phase was discarded, 12ml of a 10% sodium hydroxide solution was added to the organic phase, the pH of the solution was adjusted to 8.13, the solvent was distilled off in vacuo, 6.5ml of 6M hydrochloric acid was added to adjust the pH of the solution to 3.76, and the mixture was filtered to obtain about 14.3g of 24-demethyl-7-ketolithocholic acid.
d. Preparation of 24-norursodeoxycholic acid
Dissolving 24-demethyl-7-ketolithocholic acid in 200ml butyl acetate, adding Nicotinamide Adenine Dinucleotide Phosphate (NADP)50mg, L-sodium malate 10g, Malate Dehydrogenase (MDH)18000U, 7-beta hydroxysteroid dehydrogenase (7-beta HSDH)30000U, reacting for 3h, and sampling to determine that the residue of 24-demethyl-7-ketolithocholic acid is 0.047%. Heating the reaction solution to 70-75 ℃, stirring for 2h to denature the enzyme in the reaction solution, cooling to 30 ℃, adding 5ml of 6M hydrochloric acid, adjusting the pH value of the solution to 4.26, and filtering to remove the enzyme. Standing the filtrate for layering, discarding an aqueous phase, adding 12ml of 10% sodium hydroxide solution into an organic phase, adjusting the pH of the solution to 8.27, removing the solvent by vacuum distillation, adding 6.5ml of 6M hydrochloric acid to adjust the pH of the solution to 3.46, filtering, washing a filter cake with a large amount of water until the pH of washing water is neutral, and drying to obtain about 8.5g of 24-norursodeoxycholic acid.
Example 2
a. Preparation of 24-demethylaldehyde
Adding 200mL of dichloromethane and 20g of fully crushed seal cholic acid into a 500mL reaction bottle to obtain a micro-turbid liquid, mixing the micro-turbid liquid with 45mL of 1.0N sodium hypobromite (prepared from bromine and sodium hydroxide) in a micro mixer, and then feeding the mixture into a microchannel reactor, wherein the temperature of the microchannel reactor is 10 ℃, the pressure of the microchannel reactor is 0.2MPa, the reactor adopts a stainless steel tube of 1/16 inches, the volume of the reactor is 1mL, the reaction time is 2-5 min, and the content of seal cholic acid residue is preferably less than 0.5% by sampling after the reaction. Adding a proper amount of sodium hydrosulfite into the reaction solution to quench excessive sodium hypobromite, standing for layering, and removing a water phase; adding 200ml of saturated sodium bicarbonate solution into the dichloromethane phase, stirring for 30min, standing for layering, and removing the water phase; the dichloromethane phase was washed once more with 200ml of saturated sodium bicarbonate solution, and the aqueous phase was discarded. The methylene chloride phase was vacuum distilled to remove the distillate to give about 18.4g of a white solid (24-demethylaldehyde).
b. Preparation of 24-demethylchenodeoxycholic acid
And (3) completely dissolving 24-demethyl aldehyde in 184ml of toluene, slowly adding 40ml of 15% sodium chlorite solution in a flowing manner, continuously stirring and reacting for 1h after the sodium chlorite is added, and sampling to detect that no substrate remains. Standing for layering, and removing a water phase; adding 200ml of saturated sodium chloride solution into the toluene phase, stirring for 30min, standing for layering, and removing the water phase; adding 200ml of saturated sodium chloride solution into the toluene phase, repeatedly washing once according to the steps, and discarding the water phase; vacuum distilling toluene phase until distillate flow out to obtain white solid, dissolving the white solid in ethyl acetate, separating by column chromatography, collecting solvent segment containing 24-demethyl chenodeoxycholic acid, vacuum distilling until flow out to obtain about 10.3g of 24-demethyl chenodeoxycholic acid.
c. Preparation of 24-demethyl-7-ketolithocholic acid
Dissolving the 24-demethyl chenodeoxycholic acid in 100ml of n-butanol, adding 30mg of Nicotinamide Adenine Dinucleotide Phosphate (NADP), 5g of sodium pyruvate, 10000U of Lactate Dehydrogenase (LDH) and 5000U of 7-alpha hydroxyl steroid dehydrogenase (7-alpha HSDH), reacting for 6 hours, sampling and measuring that the residue of the 24-demethyl chenodeoxycholic acid is 0.743 percent. Heating the reaction solution to 70-75 ℃, stirring for 2h to denature the enzyme in the reaction solution, cooling to 30 ℃, adding 5ml of 6M hydrochloric acid, adjusting the pH of the solution to 4.42, filtering to remove the enzyme, standing and layering the filtrate, discarding the water phase, adding 12ml of 10% sodium hydroxide solution into the organic phase, adjusting the pH of the solution to 8.37, removing the solvent by vacuum distillation, adding 6.5ml of 6M hydrochloric acid, adjusting the pH of the solution to 3.58, and filtering to obtain about 14.6g of 24-demethyl-7-ketolithocholic acid.
d. Preparation of 24-norursodeoxycholic acid
Dissolving 24-demethyl-7-ketolithocholic acid in 100ml of n-butanol, adding Nicotinamide Adenine Dinucleotide Phosphate (NADP)30mg, L-sodium malate 7g, Malate Dehydrogenase (MDH)9000U, 7-beta hydroxysteroid dehydrogenase (7-beta HSDH)10000U, reacting for 3h, and sampling to determine that the residue of 24-demethyl-7-ketolithocholic acid is 0.321%. Heating the reaction solution to 70-75 ℃, stirring for 2h to denature the enzyme in the reaction solution, cooling to 30 ℃, adding 5ml of 6M hydrochloric acid, adjusting the pH value of the solution to 4.33, and filtering to remove the enzyme. Standing the filtrate for layering, discarding an aqueous phase, adding 12ml of 10% sodium hydroxide solution into an organic phase, adjusting the pH of the solution to 8.06, removing the solvent by vacuum distillation, adding 6.5ml of 6M hydrochloric acid to adjust the pH of the solution to 3.74, filtering, washing a filter cake with a large amount of water until the pH of washing water is neutral, and drying to obtain about 8.7g of 24-norursodeoxycholic acid.
Example 3
a. Preparation of 24-demethylaldehyde
Adding 200mL of acetone and 20g of fully crushed seal cholic acid into a 500mL reaction bottle, mixing the obtained micro-turbid liquid with 30mL of sodium hypochlorite with the concentration of 1.5N in a micro mixer, and then feeding the mixture into a micro-channel reactor, wherein the temperature of the micro-channel reactor is 10 ℃, the pressure of the micro-channel reactor is 0.2MPa, the reactor is a 1/16-inch stainless steel tube, the volume of the reactor is 1mL, the reaction time is 2-5 min, and the content of seal cholic acid residues is preferably less than 0.5% by sampling after the reaction. Adding a proper amount of sodium hydrosulfite into the reaction solution to quench excessive sodium hypochlorite, standing for layering, and discarding the water phase; adding 200ml saturated sodium bicarbonate solution into acetone phase, stirring for 30min, standing for layering, and discarding water phase; the acetone phase was washed once more with 200ml of saturated sodium bicarbonate solution, and the aqueous phase was discarded. The acetone phase was vacuum distilled to remove distillate and dried to give about 17.8g of a white solid (24-demethylaldehyde).
b. Preparation of 24-demethylchenodeoxycholic acid
And (3) completely dissolving 24-demethyl aldehyde in 100ml of methyl isobutyl ketone, slowly adding 35ml of 15% sodium perchlorate solution in a flowing manner, continuously stirring and reacting for 1h after the sodium perchlorate is added, and sampling to detect that no substrate residue exists. Standing for layering, and removing a water phase; adding 100ml saturated sodium chloride solution into the methyl isobutyl ketone phase, stirring for 30min, standing for layering, and removing the water phase; the methyl isobutyl ketone phase was washed once more with 100ml of saturated sodium chloride solution and the aqueous phase was discarded. Methyl isobutyl ketone phase was vacuum distilled until the distillate was cut off to obtain a white solid. Dissolving the white solid in ethyl acetate, separating by column chromatography, collecting solvent segment containing 24-demethyl chenodeoxycholic acid, and vacuum distilling until cut-off to obtain about 9.6g of 24-demethyl chenodeoxycholic acid.
c. Preparation of 24-demethyl-7-ketolithocholic acid
Dissolving 24-demethylchenodeoxycholic acid in 50ml of isoamyl alcohol, adding 20mg of Nicotinamide Adenine Dinucleotide Phosphate (NADP), 3g of sodium pyruvate, 5000U of Lactate Dehydrogenase (LDH) and 2000U of 7-alpha hydroxyl steroid dehydrogenase (7-alpha HSDH), reacting for 6 hours, and sampling to determine that the residue of 24-demethylchenodeoxycholic acid is 0.876%. Heating the reaction solution to 70-75 ℃, stirring for 2h to denature the enzyme in the reaction solution, cooling to 30 ℃, adding 5ml of 6M hydrochloric acid, adjusting the pH value of the solution to 4.47, and filtering to remove the enzyme. The filtrate was allowed to stand for separation, the aqueous phase was discarded, 12ml of a 10% sodium hydroxide solution was added to the organic phase, the pH of the solution was adjusted to 8.26, the solvent was distilled off in vacuo, 6.5ml of 6M hydrochloric acid was added to adjust the pH of the solution to 3.74, and the mixture was filtered to obtain about 13.8g of 24-demethyl-7-ketolithocholic acid.
d. Preparation of 24-norursodeoxycholic acid
Dissolving 24-demethyl-7-ketolithocholic acid in 50ml of isoamyl alcohol, adding Nicotinamide Adenine Dinucleotide Phosphate (NADP)20mg, L-sodium malate 5g, Malate Dehydrogenase (MDH)1500U, 7-beta hydroxysteroid dehydrogenase (7-beta HSDH)2000U, reacting for 3h, and sampling to determine that the residue of 24-demethyl-7-ketolithocholic acid is 0.685%. Heating the reaction solution to 70-75 ℃, stirring for 2h to denature the enzyme in the reaction solution, cooling to 30 ℃, adding 5ml of 6M hydrochloric acid, adjusting the pH value of the solution to 4.21, and filtering to remove the enzyme. Standing the filtrate for layering, discarding an aqueous phase, adding 12ml of 10% sodium hydroxide solution into an organic phase, adjusting the pH of the solution to 8.46, removing the solvent by vacuum distillation, adding 6.5ml of 6M hydrochloric acid to adjust the pH of the solution to 3.72, filtering, washing a filter cake with a large amount of water until the pH of washing water is neutral, and drying to obtain about 8.2g of 24-norursodeoxycholic acid.
Example 4
a. Preparation of 24-demethylaldehyde
Adding 250mL of tert-amyl alcohol and 20g of fully crushed seal cholic acid into a 500mL reaction bottle, mixing the obtained micro-turbid liquid and 35mL of 1.5N sodium hypochlorite in a micro mixer, and then feeding the mixture into a microchannel reactor, wherein the temperature of the microchannel reactor is 0 ℃, the pressure of the microchannel reactor is 0.2MPa, the reactor is an 1/16-inch stainless steel tube, the volume of the reactor is 1mL, the reaction time is 2-5 min, and the content of seal cholic acid residue is preferably less than 0.5% by sampling after the reaction. Adding a proper amount of sodium hydrosulfite into the reaction solution to quench excessive sodium hypochlorite, standing for layering, and discarding the water phase; adding 300ml saturated sodium bicarbonate solution into the tertiary amyl alcohol phase, stirring for 30min, standing for layering, and removing the water phase; the tert-amyl alcohol phase was washed once more with a further 300ml of saturated sodium bicarbonate solution, and the aqueous phase was discarded. The tert-amyl alcohol phase was vacuum distilled off to remove distillate and dried to give about 18.0g of a white solid (24-demethylaldehyde).
b. Preparation of 24-demethylchenodeoxycholic acid
24-demethyl aldehyde is dissolved in 150ml of tertiary amyl alcohol, 35ml of 15% sodium chlorite solution is slowly added in a flowing mode, after the sodium chlorite is added, the stirring reaction is continuously carried out for 1h, and a sample is taken to detect that no substrate is left. Standing for layering, and removing a water phase; adding 100ml saturated sodium chloride solution into the tertiary amyl alcohol phase, stirring for 30min, standing for layering, and removing the water phase; the tert-amyl alcohol phase was washed once more with 100ml of saturated sodium chloride solution and the aqueous phase was discarded. The tert-amyl alcohol phase was vacuum distilled until the distillate was cut off to give a white solid. Dissolving the white solid in ethyl acetate, separating by column chromatography, collecting solvent segment containing 24-demethyl chenodeoxycholic acid, and vacuum distilling until cut-off to obtain about 9.8g of 24-demethyl chenodeoxycholic acid.
c. Preparation of 24-demethyl-7-ketolithocholic acid
Dissolving 24-demethyl chenodeoxycholic acid in 100ml of tertiary amyl alcohol, cooling to-10 ℃, slowly adding 5g of bromine, continuing to react for 2 hours, sampling to measure that the residue of the 24-demethyl chenodeoxycholic acid is 1.586 percent, adding a small amount of sodium bisulfite to quench excessive bromine, standing for layering, discarding a water phase, washing an organic phase twice with 200ml of water, discarding a water phase, carrying out vacuum distillation on the organic phase to remove water in the organic phase, cooling to 5 ℃, filtering, and carrying out vacuum drying to obtain about 8.5g of 24-demethyl-7-ketolithocholic acid.
d. Preparation of 24-norursodeoxycholic acid
Dissolving 24-demethyl-7-ketolithocholic acid in 100ml sec-butyl alcohol, heating to 100 ℃ for dissolution, adding 5g of metallic sodium, maintaining reflux reaction for 2h, and sampling to measure that the residue of 24-demethyl-7-ketolithocholic acid is 0.132%; cooling to below 35 ℃, adding 45ml of 6M hydrochloric acid, measuring the pH value of the solution to be 4.36, standing for layering, and discarding the water phase; adding 200ml of water into the sec-butyl alcohol phase, washing twice, discarding the water phase, carrying out vacuum distillation on the organic phase, collecting about 65ml of distillate, separating out a small amount of crystals in the solution, cooling to 0 ℃, filtering, and drying to obtain about 6.7g of 24-norursodeoxycholic acid.
Claims (12)
1. A method for synthesizing 24-norursodeoxycholic acid is characterized by comprising the following steps:
a. dissolving a compound A in a first solvent, and oxidizing the compound A in a microchannel reactor by an oxidant 1 to obtain a compound B; adding a reducing agent to quench the reaction, standing for layering, discarding the water phase, washing the organic phase with a saturated sodium bicarbonate solution, removing the solvent, and drying to obtain a solid compound B;
b. dissolving the solid compound B in a second solvent, and oxidizing by using an oxidant 2 to obtain a compound C; adding a reducing agent for quenching reaction, standing for layering, removing a water phase, washing an organic phase by using a saturated sodium chloride solution, removing a solvent to obtain a white solid, dissolving the white solid in ethyl acetate, performing column chromatography separation, and distilling to remove the ethyl acetate to obtain a solid compound C;
c. dissolving the solid compound C in a third solvent, converting the solid compound C into a compound D through oxidation reaction, and purifying and drying the compound D to obtain a solid compound D;
the oxidation reaction is as follows: dissolving the solid compound C in a third solvent, and oxidizing by an oxidant 3 or an enzyme component 1 to obtain a compound D;
d. dissolving the solid compound D in a third solvent, and converting the solid compound D into a compound E through a reduction reaction; purifying and drying the obtained compound E;
the reduction reaction is as follows: dissolving the solid compound D in a third solvent, and reducing by using a reducing agent 1 or an enzyme component 2 to obtain a compound E;
wherein, the reaction process of the synthesis method is shown as a scheme (I):
2. the method of synthesizing 24-norursodeoxycholic acid according to claim 1, wherein: in the step a, the first solvent is selected from one of acetonitrile, an ester solvent, a ketone solvent, an alcohol solvent and chloroalkane, and the volume weight ratio of the first solvent to the compound A is 3-50 ml:1 g; the oxidant 1 is one or more of chlorine, hypochlorous acid, hypochlorite and hypobromite, or one or more of equivalents obtained through simple reaction, the molar ratio of the oxidant 1 to the compound A is 1-2.5: 1, and the oxidation reaction temperature is-10-50 ℃.
3. The method of synthesizing 24-norursodeoxycholic acid according to claim 1, wherein: in the step B, the second solvent is selected from any one of acetonitrile, an ester solvent, a ketone solvent and an alcohol solvent, and the volume weight ratio of the second solvent to the compound B is 3-20 ml:1 g; the oxidant 2 is any one or more of active nonmetal, high-valence metal oxide and acid or salt thereof, peroxide and acid or salt of nonmetal oxide, the molar ratio of the oxidant 2 to the compound B is 1-3: 1, and the oxidation reaction temperature is 5-40 ℃.
4. The method of synthesizing 24-norursodeoxycholic acid according to claim 1, wherein: in the step C, the third solvent is selected from any one of an ester solvent, a ketone solvent and an alcohol solvent, and the volume-to-weight ratio of the third solvent to the compound C is 5-20 ml:1 g.
5. The method for synthesizing 24-norursodeoxycholic acid according to claim 1, wherein in step C, the steps of oxidizing and purifying the solid compound C with the oxidant 3 to obtain the solid compound D are as follows:
dissolving the solid compound C in a third solvent, and adding an oxidant 3 to oxidize the compound C into a compound D; adding a reducing agent to quench the reaction, standing for layering, discarding the water phase, washing the organic phase with water, carrying out vacuum distillation to remove water in the organic phase, cooling for crystallization, filtering and drying to obtain a solid compound D.
6. The method of synthesizing 24-norursodeoxycholic acid according to claim 5, wherein: in the step C, the oxidant 3 is any one of active nonmetal, high-valence metal oxide and acid and salt thereof, and peroxide and acid and salt of nonmetal oxide, the molar ratio of the oxidant 3 to the compound C is 1-3: 1, and the oxidation reaction temperature is-15-20 ℃.
7. The method for synthesizing 24-norursodeoxycholic acid according to claim 1, wherein in step C, the solid compound C is oxidized by enzyme component 1 and purified to obtain solid compound D, which comprises the following specific steps:
dissolving the solid compound C in a third solvent, adding an enzyme component 1, reacting for 1-12 h at the temperature of 20-40 ℃ and the pH value of the solution of 6.0-8.0, and oxidizing to obtain a compound D;
heating the reaction solution to 70-75 ℃, stirring for 2h, cooling to 30 ℃, adjusting the pH of the solution to 4-4.5 with hydrochloric acid, filtering to remove enzyme, standing and layering the filtrate, discarding the water phase, adding 10% sodium hydroxide solution into the organic phase, adjusting the pH of the solution to 8.0-8.5, removing the solvent by vacuum distillation, adjusting the pH of the solution to 3-4 with hydrochloric acid, filtering and drying to obtain a solid compound D.
8. The method of claim 7, wherein the enzyme component 1 comprises nicotinamide adenine dinucleotide phosphate, sodium pyruvate, lactate dehydrogenase, and 7-alpha hydroxysteroid dehydrogenase, wherein the content of each component is as follows:
the weight ratio of the nicotinamide adenine dinucleotide phosphate compound C is 2-5: 1000;
the weight ratio of the sodium pyruvate to the compound C is 0.3-0.8: 1;
the activity weight ratio of the lactate dehydrogenase to the compound C is 200-2000U: 1 g;
the activity weight ratio of the alpha-hydroxysteroid dehydrogenase to the compound C is 300-1000U:1 g.
9. The method for synthesizing 24-norursodeoxycholic acid according to claim 1, wherein in step D, the specific steps of reducing and purifying the solid compound D with the reducing agent 1 to obtain the solid compound E are as follows:
dissolving a solid compound D in a third solvent, and adding a reducing agent 1 to reduce the compound D into a crude compound E; and adding hydrochloric acid to adjust the pH value of the solution to 4-4.5, layering, washing an organic phase with water, distilling in vacuum until crystals are separated out, cooling, filtering and drying.
10. The method of synthesizing 24-norursodeoxycholic acid according to claim 9, wherein: in the step D, the reducing agent 1 is one of active metal, active metal hydride and non-metal simple substance, the molar ratio of the reducing agent 1 to the compound D is 1-10: 1, and the reduction reaction temperature is 10-105 ℃.
11. The method for synthesizing 24-norursodeoxycholic acid according to claim 1, wherein in step D, the specific steps of reducing and purifying solid compound D by enzyme component 2 to obtain solid compound E are as follows:
dissolving a solid compound D in a third solvent, adding an enzyme component 2, reacting for 1-10 hours at the temperature of 10-40 ℃ and the pH value of the solution of 6.0-8.0, and reducing the compound D into a compound E;
heating the reaction solution to 70-75 ℃, stirring for 2h, cooling to 30 ℃, adjusting the pH of the solution to 4-4.5 with hydrochloric acid, filtering to remove enzyme, standing and layering the filtrate, discarding the water phase, adding 10% sodium hydroxide solution into the organic phase, adjusting the pH of the solution to 8.0-8.5, removing the solvent by vacuum distillation, adjusting the pH of the solution to 3-4 with hydrochloric acid, filtering, and drying.
12. The method of claim 11, wherein the enzyme component 2 comprises nicotinamide adenine dinucleotide phosphate, sodium L-malate, malate dehydrogenase, and 7-beta hydroxysteroid dehydrogenase, wherein the contents of each component are as follows:
the weight ratio of the nicotinamide adenine dinucleotide phosphate compound D is 2-5: 1000;
the weight ratio of the L-sodium malate to the compound D is 0.5-1.0: 1;
the activity weight ratio of the malate dehydrogenase to the compound D is 150-1800U: 1 g;
the activity weight ratio of the 7-beta hydroxysteroid dehydrogenase to the compound D is 200-3000U: 1 g.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114989240A (en) * | 2022-07-15 | 2022-09-02 | 浙江共同共新医药科技有限公司 | Continuous synthesis process for steroid derivative oxidation reaction |
| WO2024083775A1 (en) | 2022-10-20 | 2024-04-25 | Chemelectiva S.R.L. | Process for the preparation of nor-ursodeoxycholic acid |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102994604A (en) * | 2012-11-21 | 2013-03-27 | 上海凯宝药业股份有限公司 | Method for preparing binding-form ursodesoxycholic acid by two-step enzymatic method |
| CN108137643A (en) * | 2015-08-07 | 2018-06-08 | 英特塞普特医药品公司 | The method for preparing bile acid and its derivative |
| CN108251491A (en) * | 2018-03-09 | 2018-07-06 | 中山百灵生物技术有限公司 | A kind of method of enzymatic method synthesis ursodesoxycholic acid |
| CN109593812A (en) * | 2018-12-29 | 2019-04-09 | 中山百灵生物技术有限公司 | A kind of method of enzymatic method synthesis 3- ketone group deoxycholic acid |
| CN111356459A (en) * | 2017-09-28 | 2020-06-30 | 福尔克博士药物有限责任公司 | Use of norursodeoxycholic acid for reducing liver fat |
| CN112813128A (en) * | 2021-01-12 | 2021-05-18 | 中山百灵生物技术股份有限公司 | Synthetic method of alloursodeoxycholic acid |
| CN113135971A (en) * | 2020-01-20 | 2021-07-20 | 成都贝诺科成生物科技有限公司 | Carbon-losing cholestanal, and preparation method and application thereof |
| CN113832122A (en) * | 2021-10-19 | 2021-12-24 | 中山百灵生物技术股份有限公司 | 7 beta-HSDH enzyme mutant and coding gene and application thereof |
-
2021
- 2021-12-27 CN CN202111617475.1A patent/CN114276401A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102994604A (en) * | 2012-11-21 | 2013-03-27 | 上海凯宝药业股份有限公司 | Method for preparing binding-form ursodesoxycholic acid by two-step enzymatic method |
| CN108137643A (en) * | 2015-08-07 | 2018-06-08 | 英特塞普特医药品公司 | The method for preparing bile acid and its derivative |
| CN111356459A (en) * | 2017-09-28 | 2020-06-30 | 福尔克博士药物有限责任公司 | Use of norursodeoxycholic acid for reducing liver fat |
| CN108251491A (en) * | 2018-03-09 | 2018-07-06 | 中山百灵生物技术有限公司 | A kind of method of enzymatic method synthesis ursodesoxycholic acid |
| CN109593812A (en) * | 2018-12-29 | 2019-04-09 | 中山百灵生物技术有限公司 | A kind of method of enzymatic method synthesis 3- ketone group deoxycholic acid |
| CN113135971A (en) * | 2020-01-20 | 2021-07-20 | 成都贝诺科成生物科技有限公司 | Carbon-losing cholestanal, and preparation method and application thereof |
| CN112813128A (en) * | 2021-01-12 | 2021-05-18 | 中山百灵生物技术股份有限公司 | Synthetic method of alloursodeoxycholic acid |
| CN113832122A (en) * | 2021-10-19 | 2021-12-24 | 中山百灵生物技术股份有限公司 | 7 beta-HSDH enzyme mutant and coding gene and application thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114989240A (en) * | 2022-07-15 | 2022-09-02 | 浙江共同共新医药科技有限公司 | Continuous synthesis process for steroid derivative oxidation reaction |
| WO2024083775A1 (en) | 2022-10-20 | 2024-04-25 | Chemelectiva S.R.L. | Process for the preparation of nor-ursodeoxycholic acid |
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