CN106831424A - The method that simvastatin ammonium salt is prepared by Lovastatin - Google Patents
The method that simvastatin ammonium salt is prepared by Lovastatin Download PDFInfo
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- CN106831424A CN106831424A CN201510877939.0A CN201510877939A CN106831424A CN 106831424 A CN106831424 A CN 106831424A CN 201510877939 A CN201510877939 A CN 201510877939A CN 106831424 A CN106831424 A CN 106831424A
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- simvastatin
- ammonium salt
- acid
- lovastatin
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- PCZOHLXUXFIOCF-UHFFFAOYSA-N Monacolin X Natural products C12C(OC(=O)C(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 PCZOHLXUXFIOCF-UHFFFAOYSA-N 0.000 title abstract description 38
- PCZOHLXUXFIOCF-BXMDZJJMSA-N lovastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 PCZOHLXUXFIOCF-BXMDZJJMSA-N 0.000 title abstract description 38
- 229960004844 lovastatin Drugs 0.000 title abstract description 38
- QLJODMDSTUBWDW-UHFFFAOYSA-N lovastatin hydroxy acid Natural products C1=CC(C)C(CCC(O)CC(O)CC(O)=O)C2C(OC(=O)C(C)CC)CC(C)C=C21 QLJODMDSTUBWDW-UHFFFAOYSA-N 0.000 title abstract description 38
- 238000000034 method Methods 0.000 title abstract description 38
- FFPDWNBTEIXJJF-OKDJMAGBSA-N (3r,5r)-7-[(1s,2s,6r,8s,8ar)-8-(2,2-dimethylbutanoyloxy)-2,6-dimethyl-1,2,6,7,8,8a-hexahydronaphthalen-1-yl]-3,5-dihydroxyheptanoic acid;azane Chemical compound [NH4+].C1=C[C@H](C)[C@H](CC[C@@H](O)C[C@@H](O)CC([O-])=O)[C@H]2[C@@H](OC(=O)C(C)(C)CC)C[C@@H](C)C=C21 FFPDWNBTEIXJJF-OKDJMAGBSA-N 0.000 title abstract description 31
- 238000006243 chemical reaction Methods 0.000 abstract description 33
- RYMZZMVNJRMUDD-UHFFFAOYSA-N SJ000286063 Natural products C12C(OC(=O)C(C)(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 RYMZZMVNJRMUDD-UHFFFAOYSA-N 0.000 abstract description 28
- 229960002855 simvastatin Drugs 0.000 abstract description 28
- RYMZZMVNJRMUDD-HGQWONQESA-N simvastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)C(C)(C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 RYMZZMVNJRMUDD-HGQWONQESA-N 0.000 abstract description 28
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 20
- 230000007062 hydrolysis Effects 0.000 abstract description 17
- XWLXKKNPFMNSFA-HGQWONQESA-N simvastatin hydroxy acid Chemical compound C1=C[C@H](C)[C@H](CC[C@@H](O)C[C@@H](O)CC(O)=O)[C@H]2[C@@H](OC(=O)C(C)(C)CC)C[C@@H](C)C=C21 XWLXKKNPFMNSFA-HGQWONQESA-N 0.000 abstract description 15
- 239000007787 solid Substances 0.000 abstract description 15
- 108700016155 Acyl transferases Proteins 0.000 abstract description 11
- 102000057234 Acyl transferases Human genes 0.000 abstract description 11
- 125000002252 acyl group Chemical group 0.000 abstract description 7
- 238000005917 acylation reaction Methods 0.000 abstract description 6
- FJQFRDAWQRBFCG-IRUSZSJRSA-N monacolin J acid Chemical compound OC(=O)C[C@H](O)C[C@H](O)CC[C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@H](O)[C@@H]21 FJQFRDAWQRBFCG-IRUSZSJRSA-N 0.000 abstract description 6
- 125000002887 hydroxy group Chemical group [H]O* 0.000 abstract description 4
- 229940121710 HMGCoA reductase inhibitor Drugs 0.000 abstract description 3
- 230000009467 reduction Effects 0.000 abstract description 3
- 238000004904 shortening Methods 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 43
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 21
- 239000007788 liquid Substances 0.000 description 21
- 239000000243 solution Substances 0.000 description 21
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 13
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 12
- 239000000047 product Substances 0.000 description 12
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 238000000605 extraction Methods 0.000 description 9
- 102000004190 Enzymes Human genes 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 8
- 235000011114 ammonium hydroxide Nutrition 0.000 description 8
- 238000001514 detection method Methods 0.000 description 8
- 238000004519 manufacturing process Methods 0.000 description 7
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 6
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 6
- 239000005457 ice water Substances 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 239000003513 alkali Substances 0.000 description 5
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 229910000029 sodium carbonate Inorganic materials 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 4
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 238000002425 crystallisation Methods 0.000 description 4
- 230000008025 crystallization Effects 0.000 description 4
- 238000004821 distillation Methods 0.000 description 4
- ZDFOBOYQVYMVCW-IRUSZSJRSA-N monacolin J Chemical compound C([C@@H]1[C@H]2[C@@H](O)C[C@H](C=C2C=C[C@@H]1C)C)C[C@@H]1C[C@@H](O)CC(=O)O1 ZDFOBOYQVYMVCW-IRUSZSJRSA-N 0.000 description 4
- ZDFOBOYQVYMVCW-UHFFFAOYSA-N monocolin J Natural products CC1C=CC2=CC(C)CC(O)C2C1CCC1CC(O)CC(=O)O1 ZDFOBOYQVYMVCW-UHFFFAOYSA-N 0.000 description 4
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 238000003556 assay Methods 0.000 description 3
- 239000013078 crystal Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 239000000413 hydrolysate Substances 0.000 description 3
- 239000007791 liquid phase Substances 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 125000003396 thiol group Chemical group [H]S* 0.000 description 3
- 238000006276 transfer reaction Methods 0.000 description 3
- 241001465318 Aspergillus terreus Species 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Natural products CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- 108010093096 Immobilized Enzymes Proteins 0.000 description 2
- 150000003863 ammonium salts Chemical class 0.000 description 2
- 229910052786 argon Inorganic materials 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 230000007073 chemical hydrolysis Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- FKRCODPIKNYEAC-UHFFFAOYSA-N ethyl propionate Chemical compound CCOC(=O)CC FKRCODPIKNYEAC-UHFFFAOYSA-N 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000012269 metabolic engineering Methods 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 229910000027 potassium carbonate Inorganic materials 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000010792 warming Methods 0.000 description 2
- 101710161460 3-oxoacyl-[acyl-carrier-protein] synthase Proteins 0.000 description 1
- KYARBIJYVGJZLB-UHFFFAOYSA-N 7-amino-4-hydroxy-2-naphthalenesulfonic acid Chemical compound OC1=CC(S(O)(=O)=O)=CC2=CC(N)=CC=C21 KYARBIJYVGJZLB-UHFFFAOYSA-N 0.000 description 1
- -1 Acyl thioester Chemical class 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 102000004286 Hydroxymethylglutaryl CoA Reductases Human genes 0.000 description 1
- 108090000895 Hydroxymethylglutaryl CoA Reductases Proteins 0.000 description 1
- 108010028554 LDL Cholesterol Proteins 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 125000003158 alcohol group Chemical group 0.000 description 1
- 239000000908 ammonium hydroxide Substances 0.000 description 1
- CQIUKKVOEOPUDV-IYSWYEEDSA-N antimycin Chemical compound OC1=C(C(O)=O)C(=O)C(C)=C2[C@H](C)[C@@H](C)OC=C21 CQIUKKVOEOPUDV-IYSWYEEDSA-N 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000006696 biosynthetic metabolic pathway Effects 0.000 description 1
- 150000001656 butanoic acid esters Chemical group 0.000 description 1
- CQIUKKVOEOPUDV-UHFFFAOYSA-N citrinine Natural products OC1=C(C(O)=O)C(=O)C(C)=C2C(C)C(C)OC=C21 CQIUKKVOEOPUDV-UHFFFAOYSA-N 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 125000005594 diketone group Chemical group 0.000 description 1
- 238000010931 ester hydrolysis Methods 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 108091008053 gene clusters Proteins 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000000055 hyoplipidemic effect Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000012265 solid product Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/28—Preparation of carboxylic acid esters by modifying the hydroxylic moiety of the ester, such modification not being an introduction of an ester group
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Pyrane Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of method that simvastatin ammonium salt is prepared by Lovastatin.The present invention
Lip river
Cut down statin and monacolin J acid is obtained after open loop and side-chain hydrolysis; then the C8 hydroxyls for the acyl moiety of acry radical donor being supplied into monacolin J acid in the presence of acyltransferase carry out acylation reaction; the acidity of Simvastatin reaction solution is adjusted again; simvastatin acid solid is separated out, finally simvastatin acid solid described simvastatin ammonium salt is obtained into after salified.The present invention can be with reactions steps, shortening operating time, simplified experimentation, pollution, improve product quality of the reduction to environment.
Description
Technical field
The present invention relates to a kind of preparation method of simvastatin ammonium salt, the method that simvastatin ammonium salt is particularly prepared by Lovastatin belongs to pharmaceutical technology field.
Background technology
Simvastatin is the hypolipidemic that Merck companies develop, and can be used to control blood cholesterol level and prevention of cardiovascular disease, and its pharmacological action is the rate-limiting enzyme -3-hydroxy-3-methylglutaryl-coenzyme A reductase for suppressing cholesterol biosynthesis as competitive inhibitor（HMG-CoA reductase）Activity, so as to reduce the biosynthesis of cholesterol.Compared with the statinses such as the Lovastatin with same dose, Simvastatin can more effectively reduce the T-CHOL and LDL-C in blood.
Merck
Company has synthesized Simvastatin by substrate of Lovastatin, and is listed in the U.S. in August, 1989, and 1994 annual sales amounts are to reach 13.69 hundred million dollars, and ranking accounts for the 5th of all best-selling drugses.From in terms of the molecular structure of Simvastatin and Lovastatin, both differ only by a methyl, and traditional production technology is to obtain Simvastatin through full chemistry synthesis as raw material using Lovastatin.
Its chemical synthesis route has two kinds：One is the side-chain hydrolysis route of Lovastatin.Simvastatin is the semi-synthetic derivative of Lovastatin, Simvastatin be on its C-8 alpha -carbon atom of butyric acid ester side chain more than Lovastatin a methyl.1980; Hoffman etc. is disclosed with Lovastatin as initiation material in patent US4444784; the side-chain hydrolysis route of chemical synthesis Simvastatin; specific chemical reaction is main to include de- Lovastatin ester hydrolysis hydroxyl protection again; again it is esterified and four steps of deprotection; but long, yield is low the time required to the route, and the accessory substance of de-esterification is more, thus is isolated and purified to product and bring adverse effect.
Two is the direct route of methylation of Lovastatin.For example in patent CN104803959 it is chemically synthesized Simvastatin using improved.Used as most process routes is used in production at present, overall yield level is 60% to 80%.The route needs the Chemical metal reagents of various expensive or danger, and in view of similarity on Lovastatin and Simvastatin structure, it is necessary to control the residual of Lovastatin, is otherwise difficult to separate both in the product.It is readily apparent that such method to be limited to step various, many consumptions of reagent type are big, and product separates the defects such as complexity, and nonproductive Simvastatin the best approach.
With going deep into that molecular biology and metabolic engineering are studied, the enzymatic clarification of Simvastatin gradually obtains the attention of academia and industrial circle.And Simvastatin technology research and production cost turned into the key factor of product competition.The related progress of enzyme process：
One is the acquisition and application of monacolin J, by LovD（Acyltransferase）After the diketone synzyme of the acyltransferase coding of gene code is interrupted, it is possible to accumulate monacolin J or obtained by Lovastatin hydrolysis.And with monacolin J as substrate, enzyme' s catalysis Simvastatin.But the monacolin J fermentation levels of existing strain are relatively low, and still difficulty realizes large-scale industrial production.Acyltransferase is a key enzyme in Lovastatin biosynthesis pathway, and the enzyme has wider Substratspezifitaet for acyl carrier, acyl group substrate and acceptor.
The esterase catalyzed conversion zone of the existing commercialization of early utilization is selectively relatively low, can still cause as the reactions steps in chemical synthesis it is complicated the problems such as.But open
Patent CN102695792, CN101490271
In CN201080043600 and CN102712678; disclose the synthesis for carrying out Simvastatin for 46kD albumen LovD and mutant using size in the gene cluster for synthesizing statin in Aspergillus terreus; different types of acry radical donor is screened simultaneously; by using overexpression LovD coli strain and can penetration cell film α-dimethyl butyryl thioesters cosubstrate, develop the Whole Cell Biocatalysis method that citrinin J acid is converted into simvastatin acid.And α-dimethyl butyryl thioesters is the integration composition of Simvastatin bioconversion, it is efficient acry radical donor disclosed in the patent of invention.Compared to the synthetic route that methylates of Lovastatin, the course of reaction of cell catalysis does not need any chemical protecting agent.Acyl thioester is selected from the group being made up of α-dimethyl butyryl-S- methyl mercaptos propionic ester (DMB-S-MMP), dimethyl butyryl-S- ehtylmercaptos propionic ester (DMB-S-EMP) and dimethyl butyryl-S- methyl mercaptos acetic acid esters (DMB-S-MTG) and dimethyl butyryl-S- methyl mercaptos butyrate (DMB-S-MMB).
In the above prior art; by the acylation reaction of enzymatic; the reaction solution for obtaining; the acry radical donor that there is enzyme and excess need to be removed by the technological means such as centrifugation or extraction; cause to be settled out simvastatin acid using hydrochloric acid regulation PH to 2 afterwards, equally cut down statin acid can be into preparing Simvastatin after depositing of ammonium salt again.Cumbersome technological operation step is needed, such as elder generation's centrifugal purification separates enzyme, and extraction removes acry radical donor, and the aqueous solution is acidified to pH2.0 with 6N HCl after, and this causes the Simvastatin of free acid form and while the problems such as DMB-S-MPA is precipitated.
Two is that fermentation method prepares Simvastatin, and with the theory of synthetic biology, using molecular biology and metabolic engineering means, gene needed for being combined in microbial body, direct fermentation thalline produces Simvastatin.CN101473040 is reported, using the Aspergillus terreus bacterial strain of the genetic engineering transformation for building, using fermentation method production Simvastatin, but the simultaneously specific raising amount of the more traditional fermentation process of the yield of undeclared Simvastatin.The summary of other documents also indicates that, metabolic regulation is carried out to microorganism to improve the method for natural products production and does not show obvious effect in the production of Simvastatin.
The content of the invention
It is an object of the present invention to provide a kind of method that simvastatin ammonium salt is prepared by Lovastatin.The present invention has the advantages that to reduce reactions steps, shortens the operating time, simplifies pollution, the improve product quality of experimentation, reduction to environment.
In order to solve the above technical problems,
The technical scheme that the present invention is provided is as follows：
The method that simvastatin ammonium salt is prepared by Lovastatin, comprises the following steps：
a
, Lovastatin is obtained monacolin J acid after open loop and side-chain hydrolysis；
b
, add acry radical donor and acyltransferase, in the presence of acyltransferase by the acyl moiety of acry radical donor be supplied to monacolin J acid C8 hydroxyls carry out acylation reaction, obtain Simvastatin reaction solution；
c
, regulation Simvastatin reaction solution it is acid, separate out simvastatin acid solid；
d
, by simvastatin acid solid it is salified after obtain simvastatin ammonium salt.
In the above-mentioned method that simvastatin ammonium salt is prepared by Lovastatin, in step a, described hydrolysis is that, by chemical method basic hydrolysis, hydrolysis temperature is 67-77 DEG C；And the hydrolysis reaction system is alcohol and water, alcohol is selected from one or more in methyl alcohol, ethanol, isopropanol；After the end of the hydrolysis, alcohol is gone using the method for revolving.
In the above-mentioned method that simvastatin ammonium salt is prepared by Lovastatin, described alkali is selected from one or more in potassium hydroxide, NaOH, sodium carbonate, potassium carbonate.
In the foregoing method that simvastatin ammonium salt is prepared by Lovastatin, in step a, described hydrolysis is that, by immobilized enzyme hydrolysis, hydrolysis temperature is 30-36 DEG C, and hydrolysis terminates to be removed using the method for filtering.
In the foregoing method that simvastatin ammonium salt is prepared by Lovastatin, in step b, described acry radical donor is selected from α-dimethyl butyryl-S-
Methyl-mercaptopropionic acid ester (DMB-S-MMP), dimethyl butyryl-S- ethyls-mercaptopropionic acid ester (DMB-S-EMP)
, dimethyl butyryl-S- methyl mercaptos acetic acid esters (DMB-S-MTG), one or more in dimethyl butyryl-S- methyl mercaptos butyrate (DMB-S-MMB).The consumption of acry radical donor and acyltransferase is to make the C8 hydroxyl reactions of monacolin J acid complete.
In the foregoing method that simvastatin ammonium salt is prepared by Lovastatin, in step b, described acyltransferase is LovD.
In the foregoing method that simvastatin ammonium salt is prepared by Lovastatin, in step b, the ph value of reaction of acylation reaction is 9-10.
PH value in above-mentioned acylation reaction is maintained with alkali, and alkali is selected from one or several in NaOH, potassium hydroxide, ammonium hydroxide, sodium carbonate, potassium carbonate.
In the foregoing method that simvastatin ammonium salt is prepared by Lovastatin, in step c, regulation Simvastatin reaction solution is acid, is to add concentration to adjust pH value to 3-4 for the hydrochloric acid solution of 1M-12M to Simvastatin reaction solution.
In the foregoing method that simvastatin ammonium salt is prepared by Lovastatin, in step c, described precipitation simvastatin acid solid, it is first to separate out simvastatin acid solid product precipitation, separation of solid and liquid is carried out with centrifugal method again and obtains simvastatin acid solid, centrifugal condition is, 0-10 DEG C, 6000-10000 rpm, 10--30min.
In the foregoing method that simvastatin ammonium salt is prepared by Lovastatin, in step d, simvastatin acid solid is salified, it is that first simvastatin acid solid is extracted in the mixed solution of methyl alcohol and ethyl acetate, it is salified to simvastatin acid extract again, extraction temperature is 0-5 DEG C, and the mixed proportion of methyl alcohol and ethyl acetate is 0-0.2：1.
It is salified to simvastatin acid extract in the foregoing method that simvastatin ammonium salt is prepared by Lovastatin, it is the methanol solution that simvastatin acid extract is added ammoniacal liquor, in 7-15 DEG C of water bath, after being sufficiently stirred for, crystallization obtains simvastatin ammonium salt.
In the above-mentioned method that simvastatin ammonium salt is prepared by Lovastatin, the methanol solution of described ammoniacal liquor, the ratio of ammoniacal liquor and methyl alcohol is 1:0-1:5.
Compared with prior art; the present invention needs not move through purifying and is directly used in acylation reaction after Lip river is cut down and is hydrolyzed into monacolin J acid; and without being centrifuged and extracted work after the reaction of enzymatic acyl group; isolating and purifying for simvastatin acid and zymotic fluid and acry radical donor directly can be reached by acidifying, finally prepare ammonium salt.Therefore compared with other preparation methods, the present invention has the advantages that to reduce reactions steps, shortens the operating time, simplifies pollution, the improve product quality of experimentation, reduction to environment.
Specific embodiment
With reference to embodiment, the present invention is further illustrated.
Embodiment 1：Alkali hydrolysis method prepares simvastatin ammonium salt with the one-step method of acyl group transfer reaction, comprises the following steps：
1）
After 12g Lovastatins are loaded into reactor, 32g isopropanols, 10g potassium hydroxide and 0.6g water are sequentially added, are passed through nitrogen or argon gas, 72 ± 0.3 DEG C are warming up to, 5h, HPLC detection raw materials ＜ 1.5% is reacted, cool to 50 ± 5 DEG C, it is concentrated under reduced pressure into no liquid to steam, adds water 12g, continues distillation to no liquid and steam, no liquid is concentrated under reduced pressure into steam, water 12g is subsequently adding, is continued distillation and is steamed to no liquid, cool to 25 ± 5 DEG C；
2）
By step 1）The chemical hydrolysis product for obtaining adjusts pH to 9.5 ± 0.5 DEG C with hydrochloric acid, adds 8.8g 2,2- dimethylbutanoyl-S- methyl propionates and 4g to contain the thalline of acyltransferase, and pH to 9.5 ± 0.5 DEG C, 25 ± 5 DEG C of temperature are maintained with 1M sodium carbonate；
3）
Every 1h to step 2）Reaction solution detection, until the ratio ＜ 2% of Lovastatin hydrolysate；
4）
By step 2）Reaction solution be placed in 0-5 DEG C of ice-water bath, and toward being added dropwise over hydrochloric acid terminating reaction in reaction solution and continuing to drop to pH 3-4, form solid-liquid two-phase；
5）
By step 4）Solid-liquid layering sample, centrifugation solid-liquid.Condition 8000rpm, 10min, 4 DEG C；Product assay and thrown aside in detection supernatant；
6）
By step 5）Gained solid, adds the mixed liquor of methyl alcohol and ethyl acetate（Methyl alcohol：Ethyl acetate=1:5）200mL, fully extraction three times, combining extraction liquid, the process is in 0-5 DEG C of ice-water bath；
7）
Toward step 6）Extract be placed in 7-15 DEG C of water bath, be added dropwise over the methanol solution of ammoniacal liquor and be 7-8 to pH or there is crystal to separate out, continue crystallization 2h, filter to obtain white solid as simvastatin ammonium salt；
8）
Liquid phase detecting step 7）The simvastatin ammonium salt of acquisition, calculated purity and conversion ratio.
Table one：The product data of the simvastatin ammonium salt of the method for embodiment 1 production（Cut down the 12g that feeds intake in Lip river）
Embodiment 2：Immobilized enzyme hydrolysis method prepares simvastatin ammonium salt with the one-step method of acyl group transfer reaction, comprises the following steps：
1）
Lovastatin open loop：The NaOH of 7.5ml absolute methanols and 10ml 6N is mixed, is weighed during 22.5g Lovastatins add the solution.45ml water is finally added again, and it is 20 DEG C that temperature is controlled on agitator.It is stirred overnight（About 15 hours）；
2）
Lovastatin is hydrolyzed：The Lovastatin open loop substrate of 30ml is added in the beaker of 100ml, PH9.5 is adjusted, is added water and is settled to 90ml or so, heat up 35 DEG C, be subsequently adding immobilised enzymes 9g, be hydrolyzed reaction, pH9.5 is controlled with diluted alkaline during hydrolysis, at 35 DEG C, reaction is overnight to completion for temperature control；
3）
By step 2）The enzymic hydrolysates for obtaining adjusts pH to 9.5 ± 0.5 DEG C with hydrochloric acid, adds 2,2- dimethyl butyryl-S- methyl propionates and the thalline containing acyltransferase, and pH to 9.5 ± 0.5 DEG C, 25 ± 5 DEG C of temperature are maintained with 1M sodium carbonate；
4）
Every 1h to step 2）Reaction solution detection, until the ratio ＜ 2% of Lovastatin hydrolysate；
5）
By step 2）Reaction solution be placed in 0-5 DEG C of ice-water bath, and toward being added dropwise over hydrochloric acid terminating reaction in reaction solution and continuing to drop to pH 3-4, form solid-liquid two-phase；
6）
By step 4）Solid-liquid layering sample, centrifugation solid-liquid.Condition 8000rpm, 10min, 4 DEG C；Product assay and thrown aside in detection supernatant；
7）
By step 5）Gained solid, adds the mixed liquor of methyl alcohol and ethyl acetate（Methyl alcohol：Ethyl acetate=1:5）200mL, fully extraction three times, combining extraction liquid, the process is in 0-5 DEG C of ice-water bath；
8）
Toward step 6）Extract be placed in 7-15 DEG C of water bath, be added dropwise over the methanol solution of ammoniacal liquor and be 7-8 to pH or there is crystal to separate out, continue crystallization 2h, filter to obtain white solid as simvastatin ammonium salt；
9）
Liquid phase detecting step 7）The simvastatin ammonium salt of acquisition, calculated purity and conversion ratio.
Embodiment 3：Alkali hydrolysis method prepares simvastatin ammonium salt with the one-step method of acyl group transfer reaction, comprises the following steps：
1
）After 3g Lovastatins are loaded into reactor, 8g isopropanols, 2.5g potassium hydroxide and 0.15g water are sequentially added, are passed through nitrogen or argon gas, be warming up to 72 ± 0.3
℃
, 5h, HPLC detection raw materials ＜ 1.5% are reacted, cool to 50 ± 5
℃
, it is concentrated under reduced pressure into no liquid and steams, water 3g is added, continue distillation and steamed to no liquid, it is concentrated under reduced pressure into no liquid and steams, water 3g is subsequently adding, continue distillation and steamed to no liquid, cool to 25 ± 5 DEG C；
2
）By step 1）The chemical hydrolysis product for obtaining adjusts pH to 9.5 ± 0.5 with hydrochloric acid
℃
, add 2.2g 2,2- dimethylbutanoyl-S- methyl propionates and 1g to contain the thalline of acyltransferase, pH to 9.5 ± 0.5 is maintained with 1M sodium carbonate
℃
, 25 ± 5 DEG C of temperature；
3）
Every 1h to step 2）Reaction solution detection, until the ratio ＜ 2% of Lovastatin hydrolysate；
4
）By step 2）Reaction solution be placed in 0-5
℃
Ice-water bath in, and toward being added dropwise over hydrochloric acid terminating reaction in reaction solution and continuing to drop to pH 3-4, form solid-liquid two-phase；
5
）By step 4）Solid-liquid layering sample, centrifugation solid-liquid.Condition 8000rpm, 10min, 4
℃
；Product assay and thrown aside in detection supernatant；
6）
By step 5）Gained solid, adds ethyl acetate 50mL, fully extraction three times, and combining extraction liquid, the process is in 0-5
℃
Ice-water bath in；
7)
Toward step 6）Extract be placed in 7-15
℃
Water bath in, be added dropwise over the methanol solution of ammoniacal liquor（Ammoniacal liquor：Methyl alcohol is 1:5) to pH is for 7-8 or has crystal to separate out, continue crystallization 2h, filter to obtain white solid as simvastatin ammonium salt；
Liquid phase detecting step 7）The simvastatin ammonium salt of acquisition, calculated purity and conversion ratio.
Claims (10)
1. the method that simvastatin ammonium salt is prepared by Lovastatin, it is characterised in that comprise the following steps：
A, Lovastatin is obtained after open loop and side-chain hydrolysis monacolin J acid；
B, addition acry radical donor and acyltransferase, the C8 hydroxyls that the acyl moiety of acry radical donor is supplied into monacolin J acid in the presence of acyltransferase carry out acylation reaction, obtain Simvastatin reaction solution；
C, regulation Simvastatin reaction solution are acid, separate out simvastatin acid solid；
D, by simvastatin acid solid it is salified after obtain simvastatin ammonium salt.
2. the method that simvastatin ammonium salt is prepared by Lovastatin according to claim 1, it is characterised in that in step a, described hydrolysis is that, by chemical method basic hydrolysis, hydrolysis temperature is 67-77 DEG C；And the hydrolysis reaction system is alcohol and water, alcohol is selected from one or more in methyl alcohol, ethanol, isopropanol；After the end of the hydrolysis, alcohol is gone using the method for revolving.
3. the method that simvastatin ammonium salt is prepared by Lovastatin according to claim 1, it is characterised in that in step a, described hydrolysis is that, by immobilized enzyme hydrolysis, hydrolysis temperature is 30-36 DEG C.
4. the method that simvastatin ammonium salt is prepared by Lovastatin according to claim 1; it is characterized in that; in step b, described acry radical donor is selected from one or more in α-dimethyl butyryl-S- methyl-mercaptopropionic acid ester, dimethyl butyryl-S- ethyls-mercaptopropionic acid ester, dimethyl butyryl-S- methyl mercaptos acetic acid esters, dimethyl butyryl-S- methyl mercapto butyrates.
5. the method that simvastatin ammonium salt is prepared by Lovastatin according to claim 1, it is characterised in that in step b, described acyltransferase is LovD.
6. the method that simvastatin ammonium salt is prepared by Lovastatin according to claim 1, it is characterised in that in step b, the ph value of reaction of acylation reaction is 9-10.
7. the method that simvastatin ammonium salt is prepared by Lovastatin according to claim 1, it is characterised in that in step c, regulation Simvastatin reaction solution is acid, is to add concentration to adjust pH value to 3-4 for the hydrochloric acid solution of 1M-12M to Simvastatin reaction solution.
8. the method that simvastatin ammonium salt is prepared by Lovastatin according to claim 1, it is characterized in that, in step c, described precipitation simvastatin acid solid, it is first to separate out simvastatin acid solid product precipitation, then separation of solid and liquid is carried out with centrifugal method to obtain simvastatin acid solid, centrifugal condition is, 0-10 DEG C, 6000-10000
Rpm, 10--30min.
9. the method that simvastatin ammonium salt is prepared by Lovastatin according to claim 1, it is characterized in that, in step d, simvastatin acid solid is salified, it is that first simvastatin acid solid is extracted in the mixed solution of methyl alcohol and ethyl acetate, salified to simvastatin acid extract again, extraction temperature is 0-5 DEG C, and the mixed proportion of methyl alcohol and ethyl acetate is 0-0.2：1.
10. the method that simvastatin ammonium salt is prepared by Lovastatin according to claim 9, it is characterized in that, it is salified to simvastatin acid extract, it is the methanol solution that simvastatin acid extract is added ammoniacal liquor, in 7-15 DEG C of water bath, after being sufficiently stirred for, crystallization obtains simvastatin ammonium salt.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1031086A (en) * | 1987-07-10 | 1989-02-15 | 麦克公司 | The alkylating method of alpha-carbon of Mai Weinuo element and analogue 8-acyl group thereof |
| CN101415833A (en) * | 2003-10-21 | 2009-04-22 | 戴弗萨公司 | Methods for making simvastatin and intermediates |
| CN102712678A (en) * | 2009-09-30 | 2012-10-03 | 科德克希思公司 | Improved LOV-D acyltransferase mediated acylation |
| CN103787881A (en) * | 2013-11-22 | 2014-05-14 | 成都摩尔生物医药有限公司 | Simvastatin ammonium salt |
-
2015
- 2015-12-04 CN CN201510877939.0A patent/CN106831424B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1031086A (en) * | 1987-07-10 | 1989-02-15 | 麦克公司 | The alkylating method of alpha-carbon of Mai Weinuo element and analogue 8-acyl group thereof |
| CN101415833A (en) * | 2003-10-21 | 2009-04-22 | 戴弗萨公司 | Methods for making simvastatin and intermediates |
| CN102712678A (en) * | 2009-09-30 | 2012-10-03 | 科德克希思公司 | Improved LOV-D acyltransferase mediated acylation |
| CN103787881A (en) * | 2013-11-22 | 2014-05-14 | 成都摩尔生物医药有限公司 | Simvastatin ammonium salt |
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