CN116478229A - Method for preparing alfasin by using breinolone - Google Patents
Method for preparing alfasin by using breinolone Download PDFInfo
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- CN116478229A CN116478229A CN202310452637.3A CN202310452637A CN116478229A CN 116478229 A CN116478229 A CN 116478229A CN 202310452637 A CN202310452637 A CN 202310452637A CN 116478229 A CN116478229 A CN 116478229A
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- 238000000034 method Methods 0.000 title claims abstract description 34
- 238000006243 chemical reaction Methods 0.000 claims abstract description 68
- 238000000855 fermentation Methods 0.000 claims abstract description 28
- 230000004151 fermentation Effects 0.000 claims abstract description 28
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims abstract description 27
- 239000003054 catalyst Substances 0.000 claims abstract description 17
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 16
- 238000010511 deprotection reaction Methods 0.000 claims abstract description 13
- DUHUCHOQIDJXAT-OLVMNOGESA-N 3-hydroxy-(3-α,5-α)-Pregnane-11,20-dione Chemical compound C([C@@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H](C(=O)C)[C@@]2(C)CC1=O DUHUCHOQIDJXAT-OLVMNOGESA-N 0.000 claims abstract description 10
- 229960003305 alfaxalone Drugs 0.000 claims abstract description 10
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 8
- 238000005805 hydroxylation reaction Methods 0.000 claims abstract description 7
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 57
- 150000001875 compounds Chemical class 0.000 claims description 56
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 33
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 25
- 239000002904 solvent Substances 0.000 claims description 23
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 claims description 18
- 125000004185 ester group Chemical group 0.000 claims description 16
- 241000235546 Rhizopus stolonifer Species 0.000 claims description 15
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 14
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 12
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 12
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 12
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 12
- 239000003513 alkali Substances 0.000 claims description 10
- 239000002585 base Substances 0.000 claims description 10
- 238000006460 hydrolysis reaction Methods 0.000 claims description 10
- FPGGTKZVZWFYPV-UHFFFAOYSA-M tetrabutylammonium fluoride Chemical compound [F-].CCCC[N+](CCCC)(CCCC)CCCC FPGGTKZVZWFYPV-UHFFFAOYSA-M 0.000 claims description 10
- LGZDNJBUAAXEMN-UHFFFAOYSA-N 1,2,2,3-tetramethyl-1-oxidopiperidin-1-ium Chemical compound CC1CCC[N+](C)([O-])C1(C)C LGZDNJBUAAXEMN-UHFFFAOYSA-N 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 8
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 8
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 8
- KPUWHANPEXNPJT-UHFFFAOYSA-N disiloxane Chemical group [SiH3]O[SiH3] KPUWHANPEXNPJT-UHFFFAOYSA-N 0.000 claims description 8
- 230000009471 action Effects 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 7
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 7
- 229960000549 4-dimethylaminophenol Drugs 0.000 claims description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- IJOOHPMOJXWVHK-UHFFFAOYSA-N chlorotrimethylsilane Chemical compound C[Si](C)(C)Cl IJOOHPMOJXWVHK-UHFFFAOYSA-N 0.000 claims description 6
- 239000007800 oxidant agent Substances 0.000 claims description 6
- 230000001590 oxidative effect Effects 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 6
- 230000035484 reaction time Effects 0.000 claims description 6
- BCNZYOJHNLTNEZ-UHFFFAOYSA-N tert-butyldimethylsilyl chloride Chemical compound CC(C)(C)[Si](C)(C)Cl BCNZYOJHNLTNEZ-UHFFFAOYSA-N 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- 235000010469 Glycine max Nutrition 0.000 claims description 5
- 244000068988 Glycine max Species 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 5
- 108010080698 Peptones Proteins 0.000 claims description 5
- 239000005708 Sodium hypochlorite Substances 0.000 claims description 5
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 235000019319 peptone Nutrition 0.000 claims description 5
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 5
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims description 5
- 239000001393 triammonium citrate Substances 0.000 claims description 5
- 235000011046 triammonium citrate Nutrition 0.000 claims description 5
- JWUJQDFVADABEY-UHFFFAOYSA-N 2-methyltetrahydrofuran Chemical compound CC1CCCO1 JWUJQDFVADABEY-UHFFFAOYSA-N 0.000 claims description 4
- KZMGYPLQYOPHEL-UHFFFAOYSA-N Boron trifluoride etherate Chemical compound FB(F)F.CCOCC KZMGYPLQYOPHEL-UHFFFAOYSA-N 0.000 claims description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 4
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical group [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 claims description 4
- -1 PDC Chemical compound 0.000 claims description 4
- 238000011081 inoculation Methods 0.000 claims 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 claims description 4
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 4
- VEPTXBCIDSFGBF-UHFFFAOYSA-M tetrabutylazanium;fluoride;trihydrate Chemical compound O.O.O.[F-].CCCC[N+](CCCC)(CCCC)CCCC VEPTXBCIDSFGBF-UHFFFAOYSA-M 0.000 claims description 4
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 claims description 4
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 3
- UQEAIHBTYFGYIE-UHFFFAOYSA-N hexamethyldisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)C UQEAIHBTYFGYIE-UHFFFAOYSA-N 0.000 claims description 3
- QOEPTMXREDTATR-UHFFFAOYSA-N 1,2,2,3-tetramethyl-1-oxidopiperidin-1-ium-4-ol Chemical compound CC1C(O)CC[N+](C)([O-])C1(C)C QOEPTMXREDTATR-UHFFFAOYSA-N 0.000 claims description 2
- JHWIEAWILPSRMU-UHFFFAOYSA-N 2-methyl-3-pyrimidin-4-ylpropanoic acid Chemical compound OC(=O)C(C)CC1=CC=NC=N1 JHWIEAWILPSRMU-UHFFFAOYSA-N 0.000 claims description 2
- DGMOBVGABMBZSB-UHFFFAOYSA-N 2-methylpropanoyl chloride Chemical compound CC(C)C(Cl)=O DGMOBVGABMBZSB-UHFFFAOYSA-N 0.000 claims description 2
- BXVSAYBZSGIURM-UHFFFAOYSA-N 2-phenoxy-4h-1,3,2$l^{5}-benzodioxaphosphinine 2-oxide Chemical compound O1CC2=CC=CC=C2OP1(=O)OC1=CC=CC=C1 BXVSAYBZSGIURM-UHFFFAOYSA-N 0.000 claims description 2
- ISULZYQDGYXDFW-UHFFFAOYSA-N 3-methylbutanoyl chloride Chemical compound CC(C)CC(Cl)=O ISULZYQDGYXDFW-UHFFFAOYSA-N 0.000 claims description 2
- 125000004172 4-methoxyphenyl group Chemical group [H]C1=C([H])C(OC([H])([H])[H])=C([H])C([H])=C1* 0.000 claims description 2
- GSNUFIFRDBKVIE-UHFFFAOYSA-N DMF Natural products CC1=CC=C(C)O1 GSNUFIFRDBKVIE-UHFFFAOYSA-N 0.000 claims description 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 2
- WGLPBDUCMAPZCE-UHFFFAOYSA-N Trioxochromium Chemical compound O=[Cr](=O)=O WGLPBDUCMAPZCE-UHFFFAOYSA-N 0.000 claims description 2
- 150000001263 acyl chlorides Chemical class 0.000 claims description 2
- 150000008064 anhydrides Chemical class 0.000 claims description 2
- MXMOTZIXVICDSD-UHFFFAOYSA-N anisoyl chloride Chemical compound COC1=CC=C(C(Cl)=O)C=C1 MXMOTZIXVICDSD-UHFFFAOYSA-N 0.000 claims description 2
- PASDCCFISLVPSO-UHFFFAOYSA-N benzoyl chloride Chemical compound ClC(=O)C1=CC=CC=C1 PASDCCFISLVPSO-UHFFFAOYSA-N 0.000 claims description 2
- 229910000423 chromium oxide Inorganic materials 0.000 claims description 2
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 claims description 2
- 229910000041 hydrogen chloride Inorganic materials 0.000 claims description 2
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 claims description 2
- 125000001972 isopentyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])C([H])([H])* 0.000 claims description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 2
- LPNYRYFBWFDTMA-UHFFFAOYSA-N potassium tert-butoxide Chemical compound [K+].CC(C)(C)[O-] LPNYRYFBWFDTMA-UHFFFAOYSA-N 0.000 claims description 2
- 239000005051 trimethylchlorosilane Substances 0.000 claims description 2
- GDOPTJXRTPNYNR-UHFFFAOYSA-N methyl-cyclopentane Natural products CC1CCCC1 GDOPTJXRTPNYNR-UHFFFAOYSA-N 0.000 claims 1
- AKFVXAJKYLBULF-UHFFFAOYSA-N tert-butyl-chlorosilyl-dimethylsilane Chemical compound CC(C)(C)[Si](C)(C)[SiH2]Cl AKFVXAJKYLBULF-UHFFFAOYSA-N 0.000 claims 1
- CZDYPVPMEAXLPK-UHFFFAOYSA-N tetramethylsilane Chemical compound C[Si](C)(C)C CZDYPVPMEAXLPK-UHFFFAOYSA-N 0.000 claims 1
- 238000002360 preparation method Methods 0.000 abstract description 23
- 239000002994 raw material Substances 0.000 abstract description 10
- 239000003814 drug Substances 0.000 abstract description 8
- 229910052739 hydrogen Inorganic materials 0.000 abstract description 5
- 239000001257 hydrogen Substances 0.000 abstract description 5
- 230000033444 hydroxylation Effects 0.000 abstract description 4
- 230000007547 defect Effects 0.000 abstract description 2
- 230000003647 oxidation Effects 0.000 abstract description 2
- 238000000746 purification Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 12
- FGTJJHCZWOVVNH-UHFFFAOYSA-N tert-butyl-[tert-butyl(dimethyl)silyl]oxy-dimethylsilane Chemical compound CC(C)(C)[Si](C)(C)O[Si](C)(C)C(C)(C)C FGTJJHCZWOVVNH-UHFFFAOYSA-N 0.000 description 10
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 9
- 238000003756 stirring Methods 0.000 description 9
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 8
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 7
- 239000012065 filter cake Substances 0.000 description 6
- 239000012071 phase Substances 0.000 description 6
- 238000001816 cooling Methods 0.000 description 5
- 238000001914 filtration Methods 0.000 description 5
- 239000012074 organic phase Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 230000003444 anaesthetic effect Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 239000013078 crystal Substances 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 238000005070 sampling Methods 0.000 description 4
- 235000017557 sodium bicarbonate Nutrition 0.000 description 4
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000000306 component Substances 0.000 description 3
- 238000001990 intravenous administration Methods 0.000 description 3
- 229960004134 propofol Drugs 0.000 description 3
- OLBCVFGFOZPWHH-UHFFFAOYSA-N propofol Chemical compound CC(C)C1=CC=CC(C(C)C)=C1O OLBCVFGFOZPWHH-UHFFFAOYSA-N 0.000 description 3
- 238000010791 quenching Methods 0.000 description 3
- 230000000171 quenching effect Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000000967 suction filtration Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- MZWRIOUCMXPLKV-RFOVXIPZSA-N 16-Dehydropregnenolone acetate Chemical compound C([C@@H]12)C[C@]3(C)C(C(C)=O)=CC[C@H]3[C@@H]1CC=C1[C@]2(C)CC[C@H](OC(=O)C)C1 MZWRIOUCMXPLKV-RFOVXIPZSA-N 0.000 description 2
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000004945 emulsification Methods 0.000 description 2
- 239000003193 general anesthetic agent Substances 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 230000000415 inactivating effect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 150000003431 steroids Chemical class 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical class [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- BIGUZPKZUCWSQP-UHFFFAOYSA-N CC(C(CC1)OC(C2=CC=CC=C2)=O)C(C)(C)[N+]1(C)[O-] Chemical compound CC(C(CC1)OC(C2=CC=CC=C2)=O)C(C)(C)[N+]1(C)[O-] BIGUZPKZUCWSQP-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 206010039897 Sedation Diseases 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- WETWJCDKMRHUPV-UHFFFAOYSA-N acetyl chloride Chemical compound CC(Cl)=O WETWJCDKMRHUPV-UHFFFAOYSA-N 0.000 description 1
- 239000012346 acetyl chloride Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 229940035674 anesthetics Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 238000010531 catalytic reduction reaction Methods 0.000 description 1
- 238000006757 chemical reactions by type Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000012533 medium component Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical group O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000000324 neuroprotective effect Effects 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000000186 progesterone Substances 0.000 description 1
- 229960003387 progesterone Drugs 0.000 description 1
- 238000004537 pulping Methods 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 230000036280 sedation Effects 0.000 description 1
- 239000000932 sedative agent Substances 0.000 description 1
- 230000001624 sedative effect Effects 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 229910000033 sodium borohydride Inorganic materials 0.000 description 1
- 239000012279 sodium borohydride Substances 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- ILMRJRBKQSSXGY-UHFFFAOYSA-N tert-butyl(dimethyl)silicon Chemical group C[Si](C)C(C)(C)C ILMRJRBKQSSXGY-UHFFFAOYSA-N 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000009901 transfer hydrogenation reaction Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J7/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of two carbon atoms
- C07J7/0005—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of two carbon atoms not substituted in position 21
- C07J7/001—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of two carbon atoms not substituted in position 21 substituted in position 20 by a keto group
- C07J7/0015—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of two carbon atoms not substituted in position 21 substituted in position 20 by a keto group not substituted in position 17 alfa
- C07J7/002—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of two carbon atoms not substituted in position 21 substituted in position 20 by a keto group not substituted in position 17 alfa not substituted in position 16
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P33/00—Preparation of steroids
- C12P33/06—Hydroxylating
- C12P33/08—Hydroxylating at 11 position
- C12P33/10—Hydroxylating at 11 position at 11 alpha-position
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/845—Rhizopus
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
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Abstract
The invention discloses a method for preparing medicine alfasin by using Burenolol, which takes Burenolol as a raw material and prepares alfasin through four steps of hydroxyl protection, fermentation hydroxylation, oxidation and deprotection. In the preparation method, the Burenolone is used as a raw material, has the advantages of 5 alpha-hydrogen and 3 alpha-hydroxyl, hydroxyl is introduced into the 11-position by biological fermentation hydroxylation in the reaction process, and chemical reagents and catalysts with higher cost are not needed in the preparation method, so that the defects of low yield, difficult purification and the like of the prepared alfasin due to the generation of 3 beta-hydroxyl isomers in the common preparation method are avoided. The invention has mild reaction conditions and simple and convenient operation, and can realize the efficient and low-cost preparation of the medicine alfaxalone.
Description
Technical Field
The invention belongs to the technical field of organic compound preparation, and relates to a method for preparing medicine alfasolone from breinolone.
Background
Alfaxalone (Alfaxalone), chemical name 5 alpha-pregnane-3 alpha-hydroxy-11, 20-one, is a progesterone without endocrine hormone activity, and its use is neuroactive steroid anesthetic, for intravenous general anesthetic and sedative. Alfasin has no accumulation when repeatedly administered, has improved safety compared with propofol, has twice as much efficacy in sedation and anesthesia as the currently leading propofol in the industry, has less blood pressure drop than propofol, and has neuroprotective effect on fetal and adult brains at normal anesthetic doses. Alfasin is widely used worldwide as an intravenous anesthetic (intravenous Anaesthetical).
WillamR.Nes et al reported the synthesis of alfasin in 1951 (NewW.R et al, J.Am.chem.Soc.1951.73.4765), but the synthesis of its substrate was relatively cumbersome and difficult, which is not conducive to industrialization, as shown in Scheme 1.
Scheme 1
The process disclosed in 1973 for the preparation of alfasin (US patent 3714352) was not easily scalable, but the preparation process was low in yield, lacks selectivity, required large-scale purification.
Bandyopadyhayaya a.k. Et al, 2010 reported that the structure of 3 a-hydroxy was built using a configuration inversion method to prepare the main structure of alfaxalone (bandyopadyhayaya.k. Et al bioorg. Med. Chem. Lett. 2010.20.6680), but in yields of only 32%, as shown in Scheme 2.
Scheme 2
In 2013, barbera slavi kova et al (barbera slavi kova et al, j.med. Chem.2013,56,2323) reported the construction of 3 a-hydroxy structures by sodium borohydride reduction, but isomer formation was still avoided, in a yield of 56%, as shown in Scheme 3.
Scheme 3
Drawbridge pharmaceuticals in 2020, a process patent WO2020006596 was published and entered china CN112384525a as shown in Scheme 4.
Scheme 4
The method utilizes the non-transfer hydrogenation to prepare the alfasin, still generates 3 beta-hydroxy isomer, and separates the alfasin through column chromatography by adding TBSCl, and the yield is 50-60%. The method has low yield, heavy metals are used in one step finally, and the synthesis efficiency is low.
In view of this, there is an urgent need in the marketplace for new methods for the rapid preparation of neuroactive steroid anesthetics, where relatively few chemical steps and high purity are desirable for use in industrialization.
The company discloses a method for preparing the medicine Burenolone by using the dehydropregnenolone acetate (CN 112501235A) in 2020, adopts the raw material of the dehydropregnenolone acetate from natural products, has single 5 alpha-configuration and high stereoselectivity of enzyme catalytic reduction of 3-keto in the reaction process, and can realize efficient and low-cost preparation of the medicine Burenolone, as shown in Scheme 5.
Scheme 5
Based on the method, the single three-dimensional configuration of 5 alpha-hydrogen and 3 alpha-hydroxyl of the breinolone is fully utilized, and the single three-dimensional configuration is taken as a raw material, so that a process route for preparing the alfasin is developed.
Disclosure of Invention
The invention aims to provide a method for preparing alfasin by using breinolone as a raw material. In the preparation method, the alfasin is prepared by taking the Burenolone as a raw material through four steps of 3-hydroxy protection, biological fermentation 11-hydroxylation, oxidization and deprotection.
The reaction process of the preparation method is shown as a reaction formula (I):
reaction type (I)
Wherein R is selected from ester group and silyl ether group.
The method comprises the following steps:
(1) Hydroxyl protection: in a first solvent, carrying out hydroxyl protection reaction on the breinolone shown in the formula (1) and a reagent for protecting hydroxyl under alkaline conditions to obtain a compound shown in the formula (2); the reaction process is shown in a reaction formula (A):
(2) Hydroxylation by biological fermentation: fermenting the compound of formula (2) obtained in the step (1) by using rhizopus nigricans RN-M246 (Feng Kui, etc., chemical and biological engineering, 2012, 29,6, 77-79) to prepare a compound of formula (3) in one step; the reaction process is shown in a reaction formula (B):
(3) Oxidation reaction: dissolving the compound of the formula (3) obtained in the step (2) in a second solvent, and performing temperature control reaction in the presence of an oxidant and a catalyst to obtain a compound of the formula (4); the reaction process is shown in a reaction formula (C):
(4) Deprotection reaction: dissolving the compound of the formula (4) obtained in the step (3) in a third solvent, and deprotecting under the condition of alkali or a catalyst to obtain an objective product alfaxalone; the reaction process is shown in a reaction formula (D):
in the step (1) of the present invention, when the hydroxyl protecting group R is an ester group, the hydroxyl protecting reaction is specifically: reacting the compound of formula (1) with a reagent for protecting hydroxyl groups in the first solvent under the action of a base to obtain the compound of formula (2).
Wherein the ester group is selected from one or more of C2-C10 linear ester groups (ethyl propyl butyl ester and the like), isobutyl ester groups, isoamyl ester groups, phenyl ester groups, p-methoxyphenyl ester groups and the like; preferably, ethyl groups are used.
Wherein, the mol ratio of the compound of the formula (1), the reagent for protecting hydroxyl and the alkali is 1 (1-4) (0.05-5); preferably, it is 1:3:0.1.
wherein the first solvent is one or more of ethyl acetate, dichloromethane, chloroform, DMF, toluene, tetrahydrofuran, 2-methyltetrahydrofuran and the like; preferably, ethyl acetate.
Wherein the base is selected from one or more of triethylamine, diisopropylethylamine, imidazole, pyridine, DMAP and the like; preferably, it is DMAP.
Wherein the reagent for protecting the hydroxyl is one or more selected from C2-C10 linear acyl chloride or linear anhydride, isobutyryl chloride, isovaleryl chloride, benzoyl chloride, p-methoxybenzoyl chloride and the like; preferably acetyl chloride or acetic anhydride.
Wherein the temperature of the reaction is 0-50 ℃; preferably 45 ℃.
Wherein the reaction time is 2-24 hours; preferably 4h.
In the step (1) of the present invention, when the hydroxyl protecting group R is a silyl ether group, the hydroxyl protecting reaction is specifically: reacting the compound of formula (1) with a reagent for protecting hydroxyl groups in the first solvent under the action of a base to obtain the compound of formula (2).
Wherein the silyl ether group is selected from one or more of trimethyl silyl ether (TMS), tert-butyl dimethyl silyl ether (TBS) and the like; preferably t-butyldimethylsilyl (TBS).
Wherein, the mol ratio of the compound of the formula (1), the reagent for protecting hydroxyl and the alkali is 1 (2-4) (4-8); preferably, it is 1:2.5:4.
wherein the first solvent is one or more of DMF, dichloromethane, chloroform, carbon tetrachloride and the like; preferably, it is dichloromethane.
Wherein the base is selected from one or more of triethylamine, diisopropylethylamine, imidazole, pyridine, DMAP and the like; preferably imidazole.
Wherein the reagent for protecting the hydroxyl is one or more selected from trimethylchlorosilane (TMSCl), tert-butyldimethylsilyl chloride (TBSCl) and the like; preferably TBSCl.
Wherein the temperature of the reaction is 0-50 ℃; preferably 25 ℃.
Wherein the reaction time is 2-24 hours; preferably 12h.
In the step (2) of the invention, the fermentation strain is Rhizopus nigricans (Rhizopus nigricans) RN-M246 which catalyzes the compound of the formula (2) to carry out 11 alpha-hydroxylation reaction, and the fermentation medium is optimized so as to be beneficial to the growth and transformation of thalli, thereby effectively preparing the compound of the formula (3).
In the step (2), the components and weight of the fermentation medium in 1L of water are 10-30g of glucose, 20-30g of peptone, 10-20g of cold pressed soybean powder, 1-5g of tri-ammonium citrate and 2-5g of dipotassium hydrogen phosphate; preferably, the components and weights of the fermentation medium are 30g of glucose, 20g of peptone, 10g of cold-pressed soybean powder, 1g of tri-ammonium citrate and 5g of dipotassium hydrogen phosphate.
In the step (2), the pH of the fermentation medium is 5.0-6.5; preferably, the pH of the fermentation medium is 6.5.
In the step (2), the inoculation amount of the rhizopus nigricans is 25% -30%; preferably, the inoculum size for rhizopus nigricans is 30%.
In the step (2), the rhizopus nigricans is cultured by conventional slant culture and seed culture to obtain fermentation bacteria.
In the step (3), the second solvent is selected from one or more of dichloromethane, chloroform, acetone, toluene, ethyl acetate, 2-methyltetrahydrofuran and the like; preferably, it is dichloromethane.
In the step (3), the oxidant is one or more selected from sodium hypochlorite, chromium oxide, PCC, PDC, sodium dichromate, potassium dichromate and the like; preferably sodium hypochlorite.
In the step (3), the catalyst is selected from one or more of tetramethylpiperidine oxide (TEMPO), 4-hydroxy-tetramethylpiperidine oxide, 4-benzoyloxy-tetramethylpiperidine oxide and the like; preferably, it is tetramethylpiperidine oxide (TEMPO).
In the step (3), the mass ratio of the compound of the formula (3), the oxidant and the catalyst is 1: (3.5-4.5): (0.03-0.05); preferably, it is 1:3.75:0.03.
in the step (3), the temperature of the oxidation reaction is 0-25 ℃; preferably at 10 ℃.
In the step (3), the time of the oxidation reaction is 2-5h; preferably 2.5h.
In the step (4), when the hydroxyl protecting group R is an ester group, the hydrolysis reaction is specifically: and (3) carrying out hydrolysis reaction on the compound shown in the formula (4) in the third solvent under the action of alkali to obtain the alfasin.
Wherein the base is selected from LiOH, KOH, naOH, t-BuOK, K 2 CO 3 One or more of, etc.; preferably, is K 2 CO 3 。
Wherein the molar ratio of the compound of formula (4) to the base is 1: (0.5-2); preferably, it is 1:1.3.
wherein the third solvent is selected from one or two of methanol, ethanol and the like; preferably, methanol.
Wherein the temperature of the hydrolysis reaction is 10-75 ℃; preferably 65 ℃.
Wherein the time of the hydrolysis reaction is 0.3-12 h; preferably 2h.
In the step (4), when the hydroxyl protecting group R is a silyl ether group, the deprotection reaction specifically includes: and (3) carrying out deprotection reaction on the compound shown in the formula (4) in the third solvent under the action of a catalyst to obtain the alfasin.
Wherein,,the catalyst is selected from tetrabutylammonium fluoride TBAF and tetrabutylammonium fluoride trihydrate TBAF . 3H 2 One or more of O, boron trifluoride diethyl etherate, acetic acid, ethyl acetate solution of hydrogen chloride, etc.; preferably TBAF . 3H 2 O。
Wherein the mass ratio of the compound of the formula (4) to the catalyst is 1: (1-6); preferably, it is 1:4.
wherein the third solvent is selected from one or two of tetrahydrofuran, water and the like; preferably tetrahydrofuran.
Wherein the temperature of the deprotection reaction is 10-75 ℃; preferably 25 ℃.
Wherein the time of the deprotection reaction is 2-48 h; preferably 24h.
In the invention, the alfa salon is prepared efficiently by combining chemical conversion and biological fermentation by utilizing the special 3 alpha-hydroxyl and 5 alpha-hydrogen steric configuration in the Burenolone structure. In the reaction process, no matter 3 alpha-hydroxyl protection, 11-hydroxyl substitution obtained by biological fermentation, hydroxyl oxidation and deprotection reaction, the reaction conditions are safe and mild, a system of strong alkali or strong acid is not involved, the configuration of 3 alpha-hydroxyl and 5 alpha-hydrogen is not influenced, the specific three-dimensional configuration is maintained, and the problems of isomer generation and difficult separation caused by the existing chemical method for reducing carbonyl or configuration inversion method are avoided.
The invention has the beneficial effects that: the 3 beta-hydroxy isomer introduced by reducing 3-keto in the existing method is overcome by taking the Burenolone as a raw material and utilizing the single configuration of 3 alpha-hydroxy and 5 alpha-hydrogen, thereby avoiding the loss caused by separation and improving the cost. The 11-hydroxy substituent is constructed by utilizing a biological fermentation method, chemical reagents and catalysts with high cost are not needed, and the defects of a plurality of wastes and harsh reaction conditions in the existing preparation method are avoided. The invention has mild reaction conditions and simple and convenient operation, and can realize the efficient, low-cost and environment-friendly preparation of the medicine alfasin.
Detailed Description
The invention will be described in further detail with reference to the following specific examples. The procedures, conditions, experimental methods, etc. for carrying out the present invention are common knowledge and common knowledge in the art, except for the following specific references, and the present invention is not particularly limited. (the results obtained under the optimized experimental conditions are shown in the examples, and the method has potential application value).
Fermentation hydroxylation HPLC detection
The fermentation broth was centrifuged at high speed, the supernatant was diluted with methanol, and the diluted solution was filtered through a 0.45 μm organic filter membrane to be used as a test sample for HPLC analysis. Chromatographic conditions: a C18 alkylsilane-bonded reverse phase column; the mobile phase is methanol-water (3:2, volume ratio); flow rate 0.8 mL/min -1 Column temperature 25 ℃; the ultraviolet detector wavelength is 242nm.
EXAMPLE 1 preparation of bacterial suspension
Adding appropriate amount of sterile water into the inclined plane of Rhizopus nigricans, washing spores, filtering with gauze, and shaking to obtain 1×10 7 ～1×10 8 Individual mL -1 Rhizopus nigricans spore suspension.
Example 2 shake flask transformation
75mL of fermentation medium and 10mmol.L were added to a 500mL baffle shake flask -1 After sterilization, sonicating, 4% of the suspension of rhizopus nigricans spores prepared in example 1 of the invention was inoculated at 28℃for 160 r.min -1 Converting for 24h under the condition, and then increasing the rotating speed to 220 r.min -1 The conversion was continued for 48h.
EXAMPLE 3 preparation of the Compound of formula (2) (acetyl protection)
Ethyl acetate (76 mL), the compound of formula (1) (6.36 g,20 mmol) and acetic anhydride (6.13 g,60 mmol) were added to a flask, the solution was stirred, DMAP (0.245 g,2 mmol) and acetic anhydride (6.13 g,60 mmol) were added, the reaction was quenched by TLC after completion of the reaction, water (20 mL) was added, ethyl acetate (100 mL) was added to extract, and the organic phase was washed with water, washed with saturated brine and concentrated under reduced pressure to give a crude compound of formula (2) (white solid 6.9g, molar yield 95.8%).
EXAMPLE 4 preparation of the Compound of formula (2) (TBS protection)
DCM (100 mL), the compound of formula (1) (6.36 g,20 mmol) was added to a flask, the solution was stirred, TBSCl (9.85 g,65.35 mmol) and imidazole (7.12 g,104.56 mmol) were added, the reaction was carried out at 25℃for 12h, after completion of the TLC monitoring reaction, water (100 mL) was added and stirred for 10min, DCM (80 mL) was extracted, the separated organic phase was washed with water and saturated NaCl, concentrated under reduced pressure, slurried with methanol/water at 25℃for 3h, and suction filtration to give the crude compound of formula (2) (8.30 g as a white solid, 96.2% molar yield).
EXAMPLE 5 preparation of the Compound of formula (3) (acetyl protection)
In 1L of water, fermenting substrate acetyl protected compound of formula (2) 10g, fermenting medium component glucose 30g, peptone 20g, cold pressed soybean powder 10g, tri-ammonium citrate 1g, dipotassium hydrogen phosphate 5g, pH 6.5. After the primary rhizopus nigricans seeds are cultured for 24 hours, the primary rhizopus nigricans seeds are inoculated into the fermentation medium according to 30 percent of inoculation amount. The fermentation tank with 30L is used for conversion, the liquid loading amount is 80 percent, the tank pressure is 0.05MPa, and the air flow rate is 40m 3 And/hr, at 28deg.C, stirring at 180rpm, converting for 72 hr, sampling, analyzing by HPLC, completely converting substrate, inactivating fermentation broth at 85deg.C, cooling to room temperature, and suction filtering. The filter cake was extracted with ethyl acetate and concentrated to give white crystals with a crude yield of 88.2% and an acetyl protected compound of formula (3) content of 80.2%.
EXAMPLE 6 preparation of the Compound of formula (3) (TBS protection)
In 1L of water, 10g of a compound of formula (2) protected by a fermentation substrate TBS, 30g of glucose, 20g of peptone, 10g of cold-pressed soybean powder, 1g of tri-ammonium citrate, 5g of dipotassium hydrogen phosphate and pH of 6.5 are used as components of a fermentation medium. After the primary rhizopus nigricans seeds are cultured for 24 hours, the primary rhizopus nigricans seeds are inoculated into the fermentation medium according to 30 percent of inoculation amount. The fermentation tank with 30L is used for conversion, the liquid loading amount is 80 percent, the tank pressure is 0.05MPa, and the air flow rate is 40m 3 And/hr, at 28deg.C, stirring at 180rpm, converting for 72 hr, sampling, analyzing by HPLC, completely converting substrate, inactivating fermentation broth at 85deg.C, cooling to room temperature, and suction filtering. The filter cake was extracted with ethyl acetate and concentrated to give white crystals with a crude yield of 90.5% and a TBS protected compound of formula (3) content of 81.5%.
EXAMPLE 7 preparation of the Compound of formula (4) (acetyl protection)
To a 500mL three-necked flask, 16g of water was added, and 2g of sodium hydrogencarbonate, 200g of methylene chloride and 40g of an acetyl-protected compound of formula (3) were added in this order with stirring, followed by cooling after stirring to complete dissolution, and when the internal temperature of the reaction solution reached 10 ℃, 1.2g of tetramethylpiperidine oxide (TEMPO) was added. Uniformly dripping 150g of 10% sodium hypochlorite solution for 1-2 h, and controlling the dripping temperature to be 6-15 ℃. After the dripping is finished, the reaction is carried out at the temperature of 10-15 ℃ for 2-5h, the reaction of the raw materials is analyzed by sampling TLC, after the reaction is finished, 30g of 20% sodium bicarbonate solution is added for quenching reaction, the reaction is stirred for 10min, the reaction is kept still for layering (the emulsification layer is separated into water phase extraction), the water phase is extracted once by methylene dichloride and stirred for 30 min, the reaction is kept still for layering, the organic phase of methylene dichloride is combined, washed three times by 80g of saturated sodium chloride solution and stirred for 10min, and the reaction is kept still for layering. Concentrating dichloromethane to a large amount of crystals under the condition of reduced pressure, then carrying out twice with water until the dichloromethane is concentrated, adding 200g of water, stirring for 20 minutes, carrying out suction filtration, drying a filter cake by hot air circulation at 75 ℃ for 6-12 hours to obtain 38.5g of the compound of the formula (4) protected by acetyl in light yellow to white powder, wherein the yield is 96.2%.
EXAMPLE 8 preparation of the Compound of formula (4) (TBS protection)
To a 500mL three-necked flask, 16g of water was added, followed by stirring by adding 2g of sodium hydrogencarbonate, 200g of methylene chloride and 40g of a TBS-protected compound of formula (3), stirring to dissolve completely, cooling, and adding 1.2g of tetramethylpiperidine oxide (TEMPO) when the reaction solution became 10 ℃. Uniformly dripping 150g of 10% sodium hypochlorite solution for 1-2 h, and controlling the dripping temperature to be 6-15 ℃. After the dripping is finished, the reaction is carried out at the temperature of 10-15 ℃ for 2-5h, the reaction of the raw materials is analyzed by sampling TLC, after the reaction is finished, 30g of 20% sodium bicarbonate solution is added for quenching reaction, the reaction is stirred for 10min, the reaction is kept still for layering (the emulsification layer is separated into water phase extraction), the water phase is extracted once by methylene dichloride and stirred for 30 min, the reaction is kept still for layering, the organic phase of methylene dichloride is combined, washed three times by 80g of saturated sodium chloride solution and stirred for 10min, and the reaction is kept still for layering. Concentrating dichloromethane to a large amount of crystals under reduced pressure, then carrying out twice with water until the dichloromethane is concentrated, adding 200g of water, stirring for 20 minutes, carrying out suction filtration, drying a filter cake by hot air circulation at 75 ℃ for 6-12 hours to obtain 38.2g of a compound of the formula (4) protected by light yellow to white-like powder TBS, and the yield is 95.5%.
EXAMPLE 9 hydrolysis of a Compound of formula (4) preparation of Afaxalone (acetyl removal)
To the flask were added methanol (43 mL) and K 2 CO 3 (2.07 g,15 mmol), N after digestion 2 Adding a compound (3.74 g,10 mmol) of the formula (4) under protection, heating to 65 ℃ for reaction for 2 hours, cooling to 25 ℃ after TLC monitoring reaction is complete, adding 2mol/L of dilute hydrochloric acid to adjust pH=7-8, evaporating methanol under reduced pressure, adding water (20 mL), stirring for 2 hours at 25 ℃, and suction filtering; water (20 mL) was added to the filter cake, and the mixture was stirred at 25℃for 2 hours, suction filtered and dried to give refined alfasin (white solid, 3.2g, molar yield 96.4%). 1 HNMR(CDCl 3 ,400MHz):δ0.57(s,3H)；1.00(s,3H)；2.10(s,3H)；2.57and 2.50(m,2H)；2.73(t,1H,J＝9.0)；4.05(m,1H)。
EXAMPLE 10 hydrolysis of a Compound of formula (4) preparation of Afaxalone (TBS removal)
Tetrahydrofuran (25 mL), the compound of formula (4) (4.46 g,10 mmol) and N after purging were placed in a flask 2 Adding TBAF under protection . 3H 2 O (15.7 g,50 mmol), after completion of the reaction at 25℃for 24h, TLC was followed by quenching with saturated ammonium chloride, steaming out tetrahydrofuran under reduced pressure, extraction with dichloromethane (150 mL) and water (120 mL), separation of the organic phase, and subsequent extraction with saturated NaHCO 3 Washing the aqueous solution, 2N dilute hydrochloric acid, water and saturated NaCl, concentrating under reduced pressure to obtain an alfaxalone crude product, adding 80% ethanol (10 mL) into a filter cake, pulping for 2h at 25 ℃, and suction-filtering to obtain refined alfaxalone (white solid 3.18g, molar yield 95.8%).
The method of the invention takes the single chiral Burenolone as the raw material to efficiently prepare the alfaxalone, and chemical reagents and catalysts with higher cost are not needed in the preparation process, thus providing a foundation for further improving the effect of the medicine alfaxalone in guaranteeing human health.
The protection of the present invention is not limited to the above embodiments. Variations and advantages that would occur to one skilled in the art are included in the invention without departing from the spirit and scope of the inventive concept, and the scope of the invention is defined by the appended claims.
Claims (10)
1. A method for preparing alfasin by using breinolone, which is characterized in that the reaction process of the method is shown as a reaction formula (I):
wherein R is selected from ester groups and silyl ether groups;
the method comprises the following specific steps:
(1) Hydroxyl protection reaction: in a first solvent, carrying out hydroxyl protection reaction on the breinolone shown in the formula (1) and a reagent for protecting hydroxyl under alkaline conditions to obtain a compound shown in the formula (2); the reaction process is shown in a reaction formula (A):
(2) Fermentation hydroxylation reaction: fermenting the compound of the formula (2) obtained in the step (1) by using rhizopus nigricans RN-M246 to prepare a compound of the formula (3) in one step; the reaction process is shown in a reaction formula (B):
(3) Oxidation reaction: dissolving the compound of the formula (3) obtained in the step (2) in a second solvent, and performing temperature control reaction in the presence of an oxidant and a catalyst to obtain a compound of the formula (4); the reaction process is shown in a reaction formula (C):
(4) Deprotection reaction: dissolving the compound of the formula (4) obtained in the step (3) in a third solvent, and deprotecting under the condition of alkali or a catalyst to obtain an objective product alfaxalone; the reaction process is shown in a reaction formula (D):
2. the method of claim 1, wherein the ester groups are selected from one or more of C2-C10 linear ester groups, isobutyl esters, isopentyl esters, phenyl ester groups, p-methoxyphenyl ester groups; and/or the silyl ether group is selected from one or two of trimethyl silyl ether TMS and tert-butyl dimethyl silyl ether TBS.
3. The method of claim 1, wherein in step (1), the hydroxy protection reaction is specifically: the compound of formula (1) reacts with a reagent for protecting hydroxyl in the solvent under the action of alkali to obtain the compound of formula (2).
4. A process according to claim 3, wherein when R is an ester group, in step (1), the first solvent is selected from one or more of ethyl acetate, dichloromethane, chloroform, DMF, toluene, tetrahydrofuran, 2-methyltetrahydrofuran; and/or the base is selected from one or more of triethylamine, diisopropylethylamine, imidazole, pyridine and DMAP; and/or the reagent for protecting hydroxyl is selected from one or more of C2-C10 linear acyl chloride or linear anhydride, isobutyryl chloride, isovaleryl chloride, benzoyl chloride and p-methoxybenzoyl chloride; and/or the mol ratio of the compound of the formula (1), the reagent for protecting hydroxyl and the alkali is 1:1-4:0.05-5; and/or, the temperature of the reaction is 0-50 ℃; and/or the reaction time is 2-24 hours; and/or, when R is a silyl ether group, in step (1), the first solvent is one or more of DMF, dichloromethane, chloroform, carbon tetrachloride; and/or the base is selected from one or more of triethylamine, diisopropylethylamine, imidazole, pyridine and DMAP; and/or the reagent for protecting the hydroxyl is selected from one or two of trimethylchlorosilane TMSCl and tert-butyldimethylsilyl chlorosilane TBSCl; and/or the mol ratio of the compound of the formula (1), the reagent for protecting hydroxyl and the alkali is 1:2-4:4-8; and/or, the temperature of the reaction is 0-50 ℃; and/or the reaction time is 2-24 h.
5. The method according to claim 1, wherein in the step (2), the components and weight of the fermentation medium are 10-30g of glucose, 20-30g of peptone, 10-20g of cold-pressed soybean powder, 1-5g of tri-ammonium citrate and 2-5g of dipotassium hydrogen phosphate; and/or the pH of the fermentation medium is 5.0-6.5; and/or, the inoculation amount of the rhizopus nigricans is 25% -30%.
6. The method of claim 1, wherein in step (3), the second solvent is selected from one or more of dichloromethane, chloroform, acetone, toluene, ethyl acetate, 2-methyltetrahydrofuran; and/or the oxidant is selected from one or more of sodium hypochlorite, chromium oxide, PCC, PDC, sodium dichromate and potassium dichromate; and/or the catalyst is selected from one or more of tetramethylpiperidine oxide TEMPO, 4-hydroxy-tetramethylpiperidine oxide and 4-benzyloxy-tetramethylpiperidine oxide; and/or the mass ratio of the compound of the formula (3), the oxidant and the catalyst is 1:3.5-4.5:0.03-0.05; and/or the temperature of the oxidation reaction is 0 ℃ to 25 ℃.
7. The method of claim 1, wherein when R is an ester group, the hydrolysis reaction in step (4) is specifically: and (3) carrying out hydrolysis reaction on the compound shown in the formula (4) in the third solvent under the action of alkali to obtain the alfasin.
8. The process according to claim 7, wherein the base is selected from LiOH, KOH, naOH, t-BuOK, K 2 CO 3 One or more of the following; and/or, the molar ratio of the compound of formula (4) to the base is 1:0.5 to 2; and/or the third solvent is selected from one or two of methanol and ethanol; and/or the temperature of the hydrolysis reaction is 10-75 ℃; and/or the hydrolysis reaction time is 0.3-12 h.
9. The method of claim 1, wherein when R is a silyl ether group, the deprotection reaction in step (4) is specifically: and (3) carrying out deprotection reaction on the compound shown in the formula (4) in the third solvent under the action of a catalyst to obtain the alfasin.
10. The process of claim 9 wherein the catalyst is selected from the group consisting of tetrabutylammonium fluoride TBAF, tetrabutylammonium fluoride trihydrate TBAF . 3H 2 One or more of O, boron trifluoride diethyl etherate, acetic acid, ethyl acetate solution of hydrogen chloride; and/or the mass ratio of the compound of the formula (4) to the catalyst is 1:1 to 6; and/or the third solvent is selected from one or two of tetrahydrofuran and water; and/or the temperature of the deprotection reaction is 10-75 ℃; and/or the deprotection reaction time is 2-48 h.
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2023
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