CN114540455B - Process for preparing chiral beta-amino acid derivatives - Google Patents
Process for preparing chiral beta-amino acid derivatives Download PDFInfo
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- CN114540455B CN114540455B CN202011331904.4A CN202011331904A CN114540455B CN 114540455 B CN114540455 B CN 114540455B CN 202011331904 A CN202011331904 A CN 202011331904A CN 114540455 B CN114540455 B CN 114540455B
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- 150000001576 beta-amino acids Chemical class 0.000 title claims abstract description 61
- 238000004519 manufacturing process Methods 0.000 title claims description 11
- 239000000758 substrate Substances 0.000 claims abstract description 115
- 239000007787 solid Substances 0.000 claims abstract description 84
- 239000002904 solvent Substances 0.000 claims abstract description 50
- 239000002245 particle Substances 0.000 claims abstract description 46
- 238000006911 enzymatic reaction Methods 0.000 claims abstract description 44
- 239000002994 raw material Substances 0.000 claims abstract description 31
- 238000000034 method Methods 0.000 claims abstract description 30
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims abstract description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 24
- 102000004190 Enzymes Human genes 0.000 claims abstract description 23
- 108090000790 Enzymes Proteins 0.000 claims abstract description 23
- 230000008569 process Effects 0.000 claims abstract description 19
- 238000002360 preparation method Methods 0.000 claims abstract description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000003054 catalyst Substances 0.000 claims abstract description 13
- 239000012046 mixed solvent Substances 0.000 claims abstract description 6
- 238000002156 mixing Methods 0.000 claims abstract description 4
- 238000006243 chemical reaction Methods 0.000 claims description 81
- 239000000243 solution Substances 0.000 claims description 62
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 42
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical group OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 30
- 125000000217 alkyl group Chemical group 0.000 claims description 29
- NGVDGCNFYWLIFO-UHFFFAOYSA-N pyridoxal 5'-phosphate Chemical compound CC1=NC=C(COP(O)(O)=O)C(C=O)=C1O NGVDGCNFYWLIFO-UHFFFAOYSA-N 0.000 claims description 26
- 235000007682 pyridoxal 5'-phosphate Nutrition 0.000 claims description 13
- 239000011589 pyridoxal 5'-phosphate Substances 0.000 claims description 13
- 229960001327 pyridoxal phosphate Drugs 0.000 claims description 13
- 102000003929 Transaminases Human genes 0.000 claims description 10
- 108090000340 Transaminases Proteins 0.000 claims description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 125000001153 fluoro group Chemical group F* 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 9
- 230000035484 reaction time Effects 0.000 claims description 8
- 239000000337 buffer salt Substances 0.000 claims description 7
- 125000001424 substituent group Chemical group 0.000 claims description 7
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- 150000003973 alkyl amines Chemical class 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
- 229910052739 hydrogen Inorganic materials 0.000 claims description 5
- 239000001257 hydrogen Substances 0.000 claims description 5
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims description 4
- 239000007983 Tris buffer Substances 0.000 claims description 4
- 239000012527 feed solution Substances 0.000 claims description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 4
- ZMJGSOSNSPKHNH-UHFFFAOYSA-N pyridoxamine 5'-phosphate Chemical compound CC1=NC=C(COP(O)(O)=O)C(CN)=C1O ZMJGSOSNSPKHNH-UHFFFAOYSA-N 0.000 claims description 4
- 239000004471 Glycine Substances 0.000 claims description 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 3
- 101710164401 Omega-aminotransferase Proteins 0.000 claims description 3
- 229960000796 barbital sodium Drugs 0.000 claims description 3
- FTOAOBMCPZCFFF-UHFFFAOYSA-N barbitone sodium Natural products CCC1(CC)C(=O)NC(=O)NC1=O FTOAOBMCPZCFFF-UHFFFAOYSA-N 0.000 claims description 3
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 3
- WHOMFKWHIQZTHY-UHFFFAOYSA-L pyridoxine 5'-phosphate(2-) Chemical compound CC1=NC=C(COP([O-])([O-])=O)C(CO)=C1O WHOMFKWHIQZTHY-UHFFFAOYSA-L 0.000 claims description 3
- RGHFKWPGWBFQLN-UHFFFAOYSA-M sodium;5,5-diethylpyrimidin-3-ide-2,4,6-trione Chemical compound [Na+].CCC1(CC)C([O-])=NC(=O)NC1=O RGHFKWPGWBFQLN-UHFFFAOYSA-M 0.000 claims description 3
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 claims 1
- 229910052731 fluorine Inorganic materials 0.000 claims 1
- 239000011737 fluorine Substances 0.000 claims 1
- 239000003960 organic solvent Substances 0.000 abstract description 12
- 230000007547 defect Effects 0.000 abstract description 7
- 125000003158 alcohol group Chemical group 0.000 abstract description 4
- 238000009776 industrial production Methods 0.000 abstract description 4
- MFFMDFFZMYYVKS-SECBINFHSA-N sitagliptin Chemical compound C([C@H](CC(=O)N1CC=2N(C(=NN=2)C(F)(F)F)CC1)N)C1=CC(F)=C(F)C=C1F MFFMDFFZMYYVKS-SECBINFHSA-N 0.000 description 58
- 229960004034 sitagliptin Drugs 0.000 description 58
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 36
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 27
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical group CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 description 26
- 235000019441 ethanol Nutrition 0.000 description 18
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- 238000001035 drying Methods 0.000 description 9
- 238000001914 filtration Methods 0.000 description 9
- 238000004537 pulping Methods 0.000 description 9
- 239000007864 aqueous solution Substances 0.000 description 8
- HQABUPZFAYXKJW-UHFFFAOYSA-N butan-1-amine Chemical compound CCCCN HQABUPZFAYXKJW-UHFFFAOYSA-N 0.000 description 8
- 238000001514 detection method Methods 0.000 description 8
- 238000004128 high performance liquid chromatography Methods 0.000 description 8
- 125000003118 aryl group Chemical group 0.000 description 7
- 230000000052 comparative effect Effects 0.000 description 7
- 238000010025 steaming Methods 0.000 description 7
- 238000003756 stirring Methods 0.000 description 7
- 125000003545 alkoxy group Chemical group 0.000 description 6
- 238000009792 diffusion process Methods 0.000 description 6
- MFFMDFFZMYYVKS-VIFPVBQESA-N (3s)-3-amino-1-[3-(trifluoromethyl)-6,8-dihydro-5h-[1,2,4]triazolo[4,3-a]pyrazin-7-yl]-4-(2,4,5-trifluorophenyl)butan-1-one Chemical compound C([C@@H](CC(=O)N1CC=2N(C(=NN=2)C(F)(F)F)CC1)N)C1=CC(F)=C(F)C=C1F MFFMDFFZMYYVKS-VIFPVBQESA-N 0.000 description 4
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 4
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 4
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 4
- 238000005054 agglomeration Methods 0.000 description 4
- 230000002776 aggregation Effects 0.000 description 4
- 150000003863 ammonium salts Chemical class 0.000 description 4
- 125000005842 heteroatom Chemical group 0.000 description 4
- 125000000623 heterocyclic group Chemical group 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- DPBLXKKOBLCELK-UHFFFAOYSA-N pentan-1-amine Chemical compound CCCCCN DPBLXKKOBLCELK-UHFFFAOYSA-N 0.000 description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 4
- 229920006395 saturated elastomer Polymers 0.000 description 4
- 229910052717 sulfur Inorganic materials 0.000 description 4
- 125000002023 trifluoromethyl group Chemical group FC(F)(F)* 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 125000004122 cyclic group Chemical group 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 125000003341 7 membered heterocyclic group Chemical group 0.000 description 2
- 101000597273 Homo sapiens PHD finger protein 11 Proteins 0.000 description 2
- 101000935642 Homo sapiens Phosphoinositide 3-kinase adapter protein 1 Proteins 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 102100025312 Protein BCAP Human genes 0.000 description 2
- 235000019270 ammonium chloride Nutrition 0.000 description 2
- VZTDIZULWFCMLS-UHFFFAOYSA-N ammonium formate Chemical group [NH4+].[O-]C=O VZTDIZULWFCMLS-UHFFFAOYSA-N 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 125000006615 aromatic heterocyclic group Chemical group 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 238000002425 crystallisation Methods 0.000 description 2
- 230000008025 crystallization Effects 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 235000019797 dipotassium phosphate Nutrition 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 229910052736 halogen Inorganic materials 0.000 description 2
- 150000002367 halogens Chemical class 0.000 description 2
- 125000001072 heteroaryl group Chemical group 0.000 description 2
- 150000002431 hydrogen Chemical group 0.000 description 2
- 125000004356 hydroxy functional group Chemical group O* 0.000 description 2
- -1 hydroxy, amino Chemical group 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 125000004433 nitrogen atom Chemical group N* 0.000 description 2
- 125000004043 oxo group Chemical group O=* 0.000 description 2
- 229940100684 pentylamine Drugs 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000001308 synthesis method Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 125000000876 trifluoromethoxy group Chemical group FC(F)(F)O* 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000005576 amination reaction Methods 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000007810 chemical reaction solvent Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 125000004093 cyano group Chemical group *C#N 0.000 description 1
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
- C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
- C12P41/006—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by reactions involving C-N bonds, e.g. nitriles, amides, hydantoins, carbamates, lactames, transamination reactions, or keto group formation from racemic mixtures
-
- 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
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
- C12P17/182—Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system
Abstract
The invention provides a preparation method of chiral beta-amino acid derivatives. The preparation method also comprises the following steps: s1, mixing a first solvent, a biological enzyme catalyst and an amino donor to form a mixed raw material solution; the first solvent is an alcohol solvent or a mixed solvent of water and an alcohol solvent; s2, adding solid particle substrate A into the mixed raw material solution in a continuous solid strand form through continuous feeding equipment to perform enzymatic reaction, so as to obtain the chiral beta-amino acid derivative. By applying the technical scheme of the invention, the defect that the substrate is dissolved in the organic solvent and then added dropwise is effectively overcome, the problem that the substrate is easy to agglomerate and separate out in the enzymatic reaction process is solved, and the chiral beta-amino acid derivative is promoted to have higher yield. Moreover, the preparation method provided by the invention is simpler and more convenient, the raw materials are easier to obtain, the repeatability is better, and the method has better industrial production prospect.
Description
Technical Field
The invention relates to the technical field of synthesis of organic compounds, in particular to a preparation method of chiral beta-amino acid derivatives.
Background
Chiral β -amino acid derivatives are an important drug, and their chiral synthesis has been attracting attention of many scientists. The synthesis methods of chiral beta amino acid derivatives are more, and the synthesis methods using enzymes as catalysts are particularly focused by those skilled in the art due to the characteristics of mild conditions, less pollution and the like. Chinese patent CN108586346B and chinese patent CN102405281B are two typical patents for the preparation of chiral β amino acid derivatives using enzyme catalysis.
In both CN108586346B and CN102405281B, it is reported that the substrate is dissolved by an organic solvent and then fed as a solution to a reaction system comprising an amino donor and an enzyme catalyst for reaction, the substrate concentration being 100g/L to 250g/L. In the field of synthesizing small molecular organic matters at present, it is widely considered that the reaction substrate is more favorable for stabilizing a reaction system in a solution dropwise adding mode. However, according to the embodiments in the above patent, a plurality of similar experiments are carried out in combination with practical situations, and it is found that the precipitation of a substrate in a reaction system is caused by the manner of dropping a substrate solution when preparing chiral β -amino acid derivatives, and then agglomeration is generated, which causes the enzymatic reaction to be difficult to completely react, and the yield of chiral β -amino acid derivatives is reduced.
Disclosure of Invention
The invention mainly aims to provide a preparation method of chiral beta-amino acid derivatives, which solves the problems that substrates are easy to separate out in a reaction system by dripping a substrate solution when the chiral beta-amino acid derivatives are prepared, and enzymatic reaction is difficult to completely react due to agglomeration, so that the yield of the beta-amino acid derivatives is reduced.
In order to achieve the above object, according to one aspect of the present invention, there is provided a method for preparing a chiral β -amino acid derivative. The chiral beta-amino acid derivative has a structure shown in the following general formula I:
in formula I, the stereogenic center marked with x has the (R) -or (S) -configuration; an enantiomeric excess of at least 70% compared to the corresponding enantiomer; z is OR 2 、SR 2 Or NR (NR) 2 R 3 The method comprises the steps of carrying out a first treatment on the surface of the R is C 1～8 Alkyl, aryl, heteroaryl, aryl-C 1～2 Alkyl or heteroaryl-C 1～2 An alkyl group; r is R 2 And R is 3 Each independently is hydrogen, C 1～8 Alkyl, aryl or aryl-C 1～2 An alkyl group; or R is 2 And R is 3 Forms together with the nitrogen atom to which they are attached an optionally substituted member selected from O, S, NH and NC 1～4 4-to 7-membered heterocyclic ring systems of other hetero atoms of the alkyl group, the heterocyclic ring being unsubstituted or independently selected from oxo, hydroxy, halogen, C 1～4 Alkoxy and C 1～4 1-3 substituents of alkyl, wherein alkyl and alkoxy are unsubstituted or substituted with 1-5 fluoro; and the heterocyclic ring system is optionally combined with a 5-to 6-membered saturated or aromatic carbocyclic ring system or contains a member selected from O, S and NC 0～4 The 5-to 6-membered saturated or aromatic heterocyclic ring system of 1-2 hetero atoms of the alkyl group being condensed, the condensed ring system being unsubstituted or substituted by a member selected from hydroxy, amino, fluoro, C 1～4 Alkyl, C 1～4 1-2 substituents for alkoxy and trifluoromethyl; wherein, the preparation method also comprises the following steps: s1, mixing a first solvent, a biological enzyme catalyst and an amino donor to form a mixed raw material solution; the first solvent is an alcohol solvent or a mixed solvent of water and an alcohol solvent; s2, adding solid particle substrate A in a continuous solid stream form into the mixed raw material solution through a continuous feeding device to perform enzymatic reaction, wherein the structural formula of the solid particle substrate A is as followsR and Z have the same meanings as defined above, giving chiral beta-amino acid derivatives.
Further, the addition rate of the solid particle substrate A is 0.5-12.0 g/s; preferably, in the process of adding the solid particle substrate A, the continuous solid flow is controlled to be a plurality of strands, and the strands are independent and do not interfere with each other; more preferably, during the addition of the solid particulate substrate a, the plurality of continuous solid streams are controlled to assume a swinging state relative to the liquid surface upon entry into the mixed feed solution.
Further, the adding time of the solid particle substrate A is 4-12 hours; preferably, the solid particulate substrate A is added for a period of time of from 6 to 10 hours.
Further, the solid particle substrate A is added in an amount of 80 to 500g per liter of the first solvent.
Further, the molar ratio of the solid particle substrate A to the amino donor is 1 (3-10).
Further, the mixed raw material solution also comprises a buffer salt system.
Further, the buffer salt system is one or more of potassium dihydrogen phosphate/dipotassium hydrogen phosphate, tris/hydrochloric acid, triethanolamine, barbital sodium/hydrochloric acid, glycine/sodium hydroxide or amino donor/hydrochloric acid.
Further, the alcohol solvent is selected from one or more of methanol, ethanol and propanol.
Further, the first solvent is a mixed solvent of an alcohol solvent and water, wherein the mass fraction of the alcohol solvent is 20-40%.
Further, the amino donor is selected from C 1～6 One or more of alkylamine, inorganic ammonium salt; preferably C 1～6 The alkylamine is isopropylamine, butylamine or pentylamine; preferably, the inorganic ammonium salt is ammonium formate, ammonium chloride or ammonium sulfate.
Further, the mixed raw material solution further comprises an amino transfer body, wherein the amino transfer body is selected from one or more of pyridoxal phosphate, pyridoxamine phosphate or pyridoxine phosphate.
Further, the biological enzyme catalyst is transaminase; the transaminase is selected from one or more of aspartate aminotransferase (AspaTs), aromatic aminotransferase (AroATs), ω -aminotransferase (ω -ATs), branched aminotransferase (BCAPs), and D-aminotransferase (DATs).
Further, in the enzymatic reaction process, the reaction temperature is 20-60 ℃, the reaction pH value is 7.0-11, and the reaction time is 18-36 h; preferably, the reaction temperature is 35-55 ℃, and the reaction pH value is 8.0-10.5.
Further, R is aryl-C 1～2 Alkyl, wherein aryl-C 1～2 Phenyl of alkyl is unsubstituted or substituted with 1-3 substituents selected from fluoro, trifluoromethyl and trifluoromethoxy; z is NR 2 R 3 ；NR 2 R 3 Is of the structure ofR 4 Is hydrogen or C which is unsubstituted or substituted by 1 to 5 fluorine atoms 1～4 An alkyl group.
By adopting the technical scheme of the invention, the defect that the substrate is dissolved in the organic solvent and then added dropwise is more effectively overcome, the technical prejudice that the solution is added dropwise to the substrate more favorably to the reaction stability is overcome, the defect that the substrate is dissolved in the organic solvent and then added dropwise is more effectively overcome by utilizing the special reaction solvent-alcohol solvent and the special feeding mode, the problem that the substrate is easy to agglomerate and separate out in the enzymatic reaction process is solved, and the chiral beta-amino acid derivative yield is promoted to be higher. Moreover, the preparation method provided by the invention is simpler and more convenient, the raw materials are easier to obtain, the repeatability is better, and the method has better industrial production prospect. The main object of the present invention is to provide a solution to the problems of the prior art.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention. In the drawings:
FIG. 1 shows a profile of a sitagliptin purity detection control prepared in example 1 of the present invention; and
fig. 2 shows a spectrum of a comparative sample of sitagliptin optical isomer detection prepared in example 1 of the present invention.
Detailed Description
It should be noted that, in the case of no conflict, the embodiments and features in the embodiments may be combined with each other. The invention will be described in detail below with reference to the drawings in connection with embodiments.
As described in the background section, in the prior art, when chiral beta-amino acid derivatives are prepared, a substrate is separated out in a reaction system by dripping a substrate solution, and then agglomeration is generated, so that the enzymatic reaction is difficult to completely react, and the yield of the beta-amino acid derivatives is reduced. In order to solve the problem, the invention provides a preparation method of a chiral beta-amino acid derivative, which has a structure shown in the following general formula I:
in formula I, the stereogenic center marked with x has the (R) -or (S) -configuration; an enantiomeric excess of at least 70% compared to the corresponding enantiomer; z is OR 2 、SR 2 Or NR (NR) 2 R 3 The method comprises the steps of carrying out a first treatment on the surface of the R is C 1～8 Alkyl, aryl, heteroaryl, aryl-C 1～2 Alkyl or heteroaryl-C 1～2 An alkyl group; promote more solid particle substrate A to be converted into chiral beta amino acid derivative, R 2 And R is 3 Each independently is hydrogen, C 1～8 Alkyl, aryl or aryl-C 1～2 An alkyl group; or R is 2 And R is 3 Forms together with the nitrogen atom to which they are attached an optionally substituted member selected from O, S, NH and NC 1～4 4-to 7-membered heterocyclic ring systems of other hetero atoms of the alkyl group, the heterocyclic ring being unsubstituted or independently selected from oxo, hydroxy, halogen, C 1～4 Alkoxy and C 1～4 1-3 substituents of alkyl, wherein alkyl and alkoxy are unsubstituted or substituted with 1-5 fluoro; and the heterocyclic ring system is optionally saturated or aromatic with 5-to 6-membersCarbocyclic ring systems or containing members selected from O, S and NC 0～4 The 5-to 6-membered saturated or aromatic heterocyclic ring system of 1-2 hetero atoms of the alkyl group being condensed, the condensed ring system being unsubstituted or being selected from hydroxy, amino, fluoro, C 1～4 Alkyl, C 1～4 1-2 substituents for alkoxy and trifluoromethyl; wherein, the preparation method also comprises the following steps: s1, mixing a first solvent, a biological enzyme catalyst and an amino donor to form a mixed raw material solution, wherein the first solvent is an alcohol solvent or a mixed solvent of water and an alcohol solvent; s2, adding solid particle substrate A in a continuous solid stream form into the mixed raw material solution through a continuous feeding device to perform enzymatic reaction, wherein the structural formula of the solid particle substrate A is as followsR and Z have the same meanings as defined above, giving chiral beta-amino acid derivatives.
Specifically, the enzymatic reaction is as follows:
according to the invention, the first solvent, the biological enzyme catalyst and the amino donor are mixed to form a mixed raw material solution, so that a more stable and uniform reaction environment is provided for the subsequent solid particle substrate A, and the subsequent enzymatic reaction is promoted to be more stable. And then adding the solid particle substrate A into the mixed raw material solution in a continuous solid stream form through a continuous feeding device to carry out enzymatic reaction. On one hand, the invention uses the alcohol solvent as the reaction solvent of the chiral beta-amino acid derivative, and compared with the condition that the solid particle substrate A is prepared into a solution and is added into the mixed raw material solution of the alcohol solvent in a dropwise manner, the invention adopts the continuous solid strand to add the solid particle substrate A, so that the diffusion speed of the solid particle substrate A after entering the alcohol solvent can be better controlled, and the enzymatic reaction can be gradually completed in the gradual diffusion process. In addition, the special surface tension of the solid particle substrate A in the reaction system of the alcohol solvent promotes the solid particle substrate A to be relatively stably kept at the position at the upper part of the reaction system, and the chiral beta-amino acid derivative of the obtained enzymatic product gradually approaches to the lower part of the alcohol solvent reaction system along with the gradual diffusion and reaction of molecules, so that the time difference and the space difference of the gradual diffusion, the reaction and the gradual diffusion of the product of the solid particle substrate A are formed, the enzymatic reaction is promoted to be more stable in the process of the enzymatic reaction, the agglomeration is not easy to occur, and the reaction yield is higher.
In a word, the preparation method of chiral beta amino acid derivative of the invention overcomes the technical prejudice that the solution dropwise adding of the substrate is more favorable for the reaction stability, utilizes the special reaction solvent-alcohol solvent to be matched with the special feeding mode, more effectively improves the defect that the substrate is dissolved in the organic solvent and then dropwise added, solves the defect that the substrate is easy to agglomerate and separate out in the enzymatic reaction process, and promotes the yield of the chiral beta amino acid derivative to be higher. Moreover, the preparation method provided by the invention is simpler and more convenient, the raw materials are easier to obtain, the repeatability is better, and the method has better industrial production prospect.
Preferably, the solid particulate substrate A is added at a rate of 0.5 to 12.0g/s; in the process of adding the solid particle substrate A, controlling the continuous solid flow into a plurality of strands, wherein the plurality of strands are independent and do not interfere with each other; more preferably, during the addition of the solid particulate substrate a, the plurality of continuous solid streams are controlled to be in a rotating state relative to the liquid surface upon entry into the mixed feed solution. Therefore, the dispersion state of the solid particle substrate A in the mixed raw material solution can be greatly increased, the solid particle substrate A is fully positioned in each direction of the upper position of the alcohol solvent, the solid particle substrate A is promoted to be better dispersed in the mixed raw material solution, the solid particle substrate A at each position is further promoted to participate in the enzymatic reaction, better distribution is formed on the space and the diffusion speed of a target product, on one hand, the enzymatic reaction can be continuously maintained to be carried out, the enzymatic reaction is more complete, the yield of chiral beta-amino acid derivatives is higher, on the other hand, the reaction products are kept at a more proper distance, and the caking problem is better solved.
In order to further improve the yield of the chiral beta-amino acid derivative, the solid particle substrate A is added for 4 to 12 hours. More preferably, the solid particulate substrate A is added for a period of time of from 6 to 10 hours.
Preferably, the solid particulate substrate A is added in an amount of 80 to 500g per liter of the first solvent. In the range, the solubility of the solid particle substrate A in the first solvent is better, the good solubility more effectively avoids the solid particle substrate A from separating out and agglomerating in the mixed raw material solution, the solid particle substrate A is enabled to be more fully contacted with the catalyst and react, the enzymatic reaction is enabled to be more complete, more solid particle substrate A is converted into the chiral beta-amino acid derivative, and the yield of the chiral beta-amino acid derivative is higher.
In order to further increase the yield of chiral beta-amino acid derivatives, the molar ratio of the solid particle substrate A to the amino donor is 1 (3-10). In this range, the reaction of the solid particulate substrate A and the amino donor is more complete, more solid particulate substrate A is converted into chiral β -amino acid derivatives, and the yield of chiral β -amino acid derivatives is higher.
In a preferred embodiment, the mixed feed solution further comprises a buffer salt system; the buffer salt system is one or more of potassium dihydrogen phosphate/dipotassium hydrogen phosphate, tris/hydrochloric acid, triethanolamine, barbital sodium/hydrochloric acid, glycine/sodium hydroxide or amino donor/hydrochloric acid. The buffer system can keep the pH value of the reaction solution system in a range more suitable for the enzymatic reaction of the invention, and promote the catalytic activity of the enzyme catalyst to be better, the stability to be better and the stereoselectivity to be higher. Meanwhile, the enzymatic reaction is more complete, and the yield of the chiral beta-amino acid derivative is higher.
Preferably, the alcohol solvent is one or more of methanol, ethanol and propanol. For the purpose of promoting better solubility of the above solid particulate substrate a in the first solvent, in a preferred embodiment, the first solvent is a mixed solvent of water and an alcohol solvent, and the mass fraction of the alcohol solvent is 20 to 40%.
In a preferred embodiment, the amino donor is preferably selected from C 1～6 One or more of alkylamine and inorganic ammonium salt can have good amination effect under lower addition amount, so as to promote more solid particlesThe substrate A is converted into chiral beta amino acid derivatives, and the yield of the chiral beta amino acid derivatives is higher. Meanwhile, based on the amino donor, carbonyl byproducts generated by the enzymatic reaction are easier to remove. More preferably C 1～6 The alkylamine is isopropylamine, butylamine or pentylamine, and the inorganic ammonium salt is ammonium formate, ammonium chloride or ammonium sulfate. The above amino donors are less costly and have the potential for large scale production.
For the purpose of delivering an amino group to the above-mentioned amino donor, it is preferable that an amino group-delivering body is further included in the mixed raw material solution, and the amino group-delivering body is selected from one or more of pyridoxal phosphate, pyridoxamine phosphate or pyridoxine phosphate.
In a preferred embodiment, the bio-enzyme catalyst is a transaminase; the transaminase is selected from one or more of aspartate aminotransferase (AspaTs), aromatic aminotransferase (AroATs), ω -aminotransferase (ω -ATs), branched aminotransferase (BCAPs), and D-aminotransferase (DATs). The enzyme has better catalytic activity and better stability.
Preferably, in the enzymatic reaction process, the reaction temperature is 20-60 ℃, the reaction pH value is 7.0-11, and the reaction time is 18-36 h; preferably, the reaction temperature is 35-55 ℃, and the reaction pH value is 8.0-10.5. Under the reaction conditions, the enzymatic reaction is more stable and complete, and the chiral beta amino acid derivative has higher yield.
In a preferred embodiment, R is aryl-C 1～2 Alkyl, wherein aryl-C 1～2 Phenyl of alkyl is unsubstituted or substituted with 1-3 substituents selected from fluoro, trifluoromethyl and trifluoromethoxy; z is NR 2 R 3 ；NR 2 R 3 Is of the structure ofR 4 Is hydrogen or C which is unsubstituted or substituted by 1 to 5 fluorine atoms 1～4 An alkyl group. For example, the chiral β -amino acid derivative is sitagliptin, having the formula +.>
The present application is described in further detail below in conjunction with specific embodiments, which should not be construed as limiting the scope of the claims.
Characterization of the properties:
the method comprises the following steps:
instrument: high performance liquid chromatograph
A detector: an ultraviolet detector; chromatographic column: discovery Cyano,150×4.6mm,5 μm; detection wavelength: 205nm; column temperature: 30 ℃; flow rate: 1.0ml/min; sample injection volume: 20. Mu.L; run time: 25min; sample injection concentration: 0.1mg/ml;
buffer salt solution: 1.36g of potassium dihydrogen phosphate, dissolved in 1000ml of water, and adjusted to ph=2.0 with phosphoric acid.
Mobile phase: buffer solution: acetonitrile=85:15
Fig. 1 shows a sitagliptin purity detection profile prepared in example 1 of the present invention.
(II) optical isomer detection method:
instrument: high performance liquid chromatograph
Chromatographic column: chiralpak AD 4.6mm by 25cm,5 μm or equivalent column; sample injection volume: 10 μl; flow rate: 0.8ml/min; column temperature: 35 ℃; detection wavelength: 268nm; run time: 20min.
Mobile phase: absolute ethyl alcohol: n-heptane: diethylamine: water = 600:400:1:1
Figure 2 shows the detection profile of the optical isomer of sitagliptin prepared in example 1 of the present invention.
Example 1
Adding a first solvent (150L of methanol and 210L of water), 60kg of isopropylamine, 100g of monopotassium phosphate and 500g of dipotassium phosphate into a reaction kettle respectively, adjusting the pH of the solution to 9.0 by using hydrochloric acid, and adding 90L of 5.0g/L (R) -omega-ATs-117 enzyme solution and 1kg of pyridoxal phosphate;
the reaction vessel was warmed to 45 ℃ and stirred while the sitagliptin substrate was added as a continuous solid stream in a swinging state relative to the liquid level. Wherein the addition rate of the sitagliptin substrate is 12g/s, the total feeding time is 10h, the addition amount of the sitagliptin substrate is 88g for each liter of the first solvent, and 4mol/L of isopropylamine aqueous solution is used for maintaining the pH value at 9.0 in the enzymatic reaction process, and the reaction time is 20h. The reaction was monitored by HPLC control and after completion of the reaction, the sitagliptin content was 87% and the corresponding (S) -configurational isomer content was 1.5%. Extracting with 500L ethyl acetate, steaming to remove organic solvent, adding 200kg toluene, pulping, filtering, and drying to obtain 35.1kg sitagliptin with 82% yield.
Wherein, the molar ratio of the sitagliptin substrate to the isopropylamine is 1:10.
example 2
Adding a first solvent (25L of methanol and 16L of water), 4kg of isopropylamine and 16L of Tris/hydrochloric acid buffer solution with pH of 8.0 into a reaction kettle respectively, adjusting the pH of the solution to 8.0 by using hydrochloric acid, and adding 9L of 2.5g/L (R) -omega-ATs-117 enzyme solution and 1kg of pyridoxal phosphate;
the reaction vessel was placed in an oil bath at 30 ℃ with stirring while the sitagliptin substrate was added in a continuous solid stream, wherein the rate of addition of the sitagliptin substrate was 2g/s and the total running-in time was 6 hours, the amount of sitagliptin substrate added was 83g per liter of the first solvent, and 4mol/L of isopropylamine aqueous solution was used to maintain pH at 8.0 during the enzymatic reaction, and the reaction time was 24. The reaction was monitored by HPLC control and after completion of the reaction, the sitagliptin content was 85% and the corresponding (S) -configurational isomer content was 1.0%. Extracting with 50L ethyl acetate, steaming to remove organic solvent, adding 20kg toluene, pulping, filtering, and drying to obtain 3.5kg sitagliptin with 80% yield.
Wherein the molar ratio of the sitagliptin substrate to the isopropylamine is 1:6.
Example 3
First solvent (50 ml methanol and 20ml water), 60g butylamine were added to the reaction flask, pH was adjusted to 8.5 with hydrochloric acid, 90ml of 5.0g/L (R) -omega-ATs-117 enzyme solution, 1g pyridoxal phosphate was added;
the reaction flask was placed in a 45 ℃ oil bath with stirring while the sitagliptin substrate was added in a continuous solid stream, wherein the rate of addition of the sitagliptin substrate was 0.5g/s and the total running-in time was 6 hours, and for each liter of first solvent, the amount of sitagliptin substrate added was 504g, and 4mol/L aqueous isopropylamine solution was used to maintain pH at 8.5 during the enzymatic reaction for 30 hours. The reaction was monitored by HPLC control and after completion of the reaction, the sitagliptin content was 80% and the corresponding (S) -configurational isomer content was 2.1%. Extracting with 500mL ethyl acetate, steaming to remove the organic solvent, adding 1kg toluene, pulping, filtering, and drying to obtain 861.8g sitagliptin with a yield of 80%.
Wherein, the mol ratio of the sitagliptin substrate to the butylamine is 1:3.3.
Example 4
Adding a first solvent (150L of methanol and 210L of water), 60kg of isopropylamine, 100g of monopotassium phosphate and 500g of dipotassium phosphate into a reaction kettle respectively, adjusting the pH of the solution to 9.0 by using hydrochloric acid, and adding 90L of 5.0g/L (R) -omega-ATs-117 enzyme solution and 1kg of pyridoxal phosphate;
the reaction vessel was warmed to 45 ℃ and stirred while the sitagliptin substrate was added as a continuous solid stream in a swinging state relative to the liquid level. Wherein the adding speed of the sitagliptin substrate is 12g/s, the total feeding time is 12h, the adding amount of the sitagliptin substrate is 103g for each liter of the first solvent, and 4mol/L isopropylamine aqueous solution is used for maintaining the pH value at 9.0 in the enzymatic reaction process, and the reaction time is 20h. The reaction was monitored by HPLC control and after completion of the reaction, the sitagliptin content was 88% and the corresponding (S) -configurational isomer content was 1.6%. Extracting with 500L ethyl acetate, steaming to remove organic solvent, adding 200kg toluene, pulping, filtering, and drying to obtain 42.9kg sitagliptin with 83% yield.
Wherein, the molar ratio of the sitagliptin substrate to the isopropylamine is 1:10.
example 5
Adding 60kg of isopropylamine into a reaction kettle, adjusting the pH of the solution to 9.0 by using hydrochloric acid, adding 90L of 10.0g/L (S) -omega-ATs-Vf enzyme solution and 1kg of pyridoxal phosphate;
the reaction vessel was warmed to 50 ℃ and stirred while the sitagliptin substrate was added as a continuous solid stream in a swinging state relative to the liquid level. Wherein the addition rate of the sitagliptin substrate is 12g/s, the total feeding time is 12h, the addition amount of the sitagliptin substrate is 103g for each liter of the first solvent, and 4mol/L of isopropylamine aqueous solution is used for maintaining the pH value at 9.0 in the enzymatic reaction process, and the reaction time is 24h. The reaction was monitored by HPLC control and after completion of the reaction, the (S) -sitagliptin content was 71% and the corresponding (R) -configurational isomer content was 3.4%. Extracting with 500L ethyl acetate, steaming to remove organic solvent, adding 100kg toluene, pulping, filtering, and drying to obtain 16.5kg (S) -sitagliptin with a yield of 32%.
Example 6
Adding 600g of isopropylamine into a reaction flask, adjusting the pH of the solution to 8.0 by using hydrochloric acid, adding 500mL of 10.0g/L (S) -omega-ATs-113 enzyme solution and 1g of pyridoxal phosphate;
the flask was warmed to 45 ℃ and stirred while the sitagliptin substrate was added as a continuous solid stream in a swinging state relative to the liquid level. Wherein the addition rate of the sitagliptin substrate is 2g/s, the total feeding time is 1h, the addition amount of the sitagliptin substrate is 180g for each liter of the first solvent, and 4mol/L of isopropylamine aqueous solution is used for maintaining the pH value at 9.0 in the enzymatic reaction process, and the reaction time is 30h. The reaction was monitored by HPLC control and after completion of the reaction, the (S) -sitagliptin content was 70% and the corresponding (R) -configurational isomer content was 4.6%. Extracting with 5L ethyl acetate, steaming to remove the organic solvent, adding 1kg toluene, pulping, filtering, and drying to obtain 275.9g (S) -sitagliptin with a yield of 38%.
Comparative example 1
50ml of methanol, 360ml of water and 60g of isopropylamine are respectively added into a reaction bottle, the pH value of the solution is adjusted to 8.5 by using hydrochloric acid, 90ml of 5g/L (R) -omega-ATs-117 enzyme solution and 1g of pyridoxal phosphate are added;
the reaction flask was placed in a 45 ℃ oil bath with stirring, while slowly adding a sitagliptin substrate solution for a total of 8 hours, wherein the sitagliptin substrate solution was dissolved in 200.0g of sitagliptin substrate in 100ml of methanol, and the pH was maintained at 8.5 during the enzymatic reaction using 4mol/L aqueous isopropylamine.
When the reaction was carried out for 6 hours, it was found that a solid precipitated in the reaction flask, and the precipitated solid was detected by HPLC, and it was confirmed that the precipitated solid was a substrate, and a caking phenomenon occurred. After completion of the reaction, the sitagliptin content was 26% and the corresponding (S) -isomer content was 96%. Because of the low content of sitagliptin, no post-treatment can be performed to obtain sitagliptin.
Comparative example 2
360ml of water and 60g of isopropylamine are respectively added into a reaction bottle, the pH value of the solution is adjusted to 8.5 by using hydrochloric acid, 90ml of 5g/L (R) -omega-ATs-117 enzyme solution and 1g of pyridoxal phosphate are added;
the reaction flask was placed in a 45 ℃ oil bath with stirring, while slowly adding a sitagliptin substrate solution for a total of 8 hours, wherein the sitagliptin substrate solution was 50ml methanol to dissolve 200.0g of sitagliptin substrate, and 4mol/L aqueous isopropylamine was used to maintain pH at 8.5 during the enzymatic reaction.
When the reaction was carried out for 4 hours, it was found that a solid precipitated in the reaction flask, and the solid was confirmed to be a substrate by HPLC detection, and a caking phenomenon occurred. After completion of the reaction, the sitagliptin content was 12% and the corresponding (S) -isomer content was 96%. Because of the low content of sitagliptin, no post-treatment can be performed to obtain sitagliptin.
Comparative example 3
100ml of DMSO,300ml of water, 60g of isopropylamine and 10g of triethanolamine are added into a 1L reaction bottle, 90ml of 2.5g/L (R) -omega-ATs-117 enzyme solution is added after the pH is regulated to 10.0 by using hydrochloric acid, 1g of pyridoxal phosphate is put into an oil bath with the temperature of 55 ℃ for stirring, 30.0g of sitagliptin substrate is slowly added, the total feeding time is 4 hours, 4mol/L of isopropylamine aqueous solution is used for maintaining the pH at about 10.0 in the feeding process, the reaction condition is monitored by central control, the sitagliptin content is 83%, and the corresponding (S) -configuration isomer content is 97%. After the reaction is completed, 1.5L of ethyl acetate is added for extraction, ethyl acetate is distilled off after layering, 1L of toluene is added for pulping and crystallization, 12.87g of sitagliptin is obtained after filtration and drying, and the yield is 43%.
Comparative example 4
50ml of DMSO and 360ml of water are added into a 1L reaction bottle, 60g of isopropylamine are added into the reaction bottle after the pH value is regulated to 8.5 by using hydrochloric acid, 90ml of 5g/L (R) -omega-ATs-117 enzyme solution and 1g of pyridoxamine phosphate are added into the reaction bottle, the mixture is put into an oil bath at 45 ℃ for stirring, 200.0g of sitagliptin substrate is slowly added, the total running-in time is 8 hours, 4mol/L of isopropylamine aqueous solution is used for maintaining the pH value to be about 8.5 in the running-in process, the reaction condition is monitored by central control, and after the reaction is completed, the sitagliptin content is 86%, and the corresponding (S) -configuration isomer content is 96%. After the reaction is completed, 1.5L of ethyl acetate is added for extraction, ethyl acetate is distilled off after layering, 1.5L of toluene is added for pulping and crystallization, 83.79g of sitagliptin is obtained after filtration and drying, and the yield is 42%.
Comparative example 5
360ml of water and 60g of isopropylamine are respectively added into a reaction bottle, the pH value of the solution is adjusted to 8.5 by using hydrochloric acid, 90ml of 5g/L (R) -omega-ATs-117 enzyme solution and 1g of pyridoxal phosphate are added;
the reaction bottle is placed in an oil bath at 45 ℃ for stirring, meanwhile, a sitagliptin substrate solution is slowly added, the total running-in time is 8 hours, wherein the sitagliptin substrate solution is 50ml of DMSO for dissolving 200.0g of sitagliptin substrate, 4mol/L of isopropylamine aqueous solution is used for maintaining the pH value in the enzymatic reaction process at 8.5, the reaction condition is monitored in a central control manner, and after the reaction is completed, the sitagliptin content is 84%, and the corresponding (S) -configuration isomer content is 97%. After the reaction is completed, adding 1L of ethyl acetate for extraction, layering, steaming to remove the ethyl acetate, adding 1L of toluene, pulping, crystallizing, filtering and drying to obtain 77.81g of sitagliptin, wherein the yield is 39%.
From the above description, it can be seen that the above embodiments of the present invention achieve the following technical effects:
as can be seen from the data of examples 1 to 4 and comparative examples 1 to 5, the present invention firstly mixes the first solvent, the bio-enzyme catalyst and the amino donor to form a mixed raw material solution, thereby providing a more stable and uniform reaction environment for the subsequent solid particle substrate a and promoting the subsequent enzymatic reaction to be more stable. And then adding the solid particle substrate A into the mixed raw material solution in a continuous solid stream form through a continuous feeding device to carry out enzymatic reaction. On the one hand, the method realizes more effective control of the content of the solid particle substrate A in the mixed raw material solution, ensures that the content change of the solid particle substrate A in the unit mixed raw material solution volume is smoother, namely, the content of the solid particle substrate A in the unit mixed raw material solution volume can be better kept to be more soluble, further ensures that the solid particle substrate A has better solubility in the mixed raw material solution, more effectively avoids the solid particle substrate A from separating out and agglomerating in the mixed raw material solution, ensures that the enzymatic reaction is more complete, and ensures that more solid particle substrate A is converted into chiral beta-amino acid derivatives, and the yield of the chiral beta-amino acid derivatives is higher. On the other hand, the contact between the solid particle substrate A and the mixed raw material solution is greatly increased, so that more solid particle substrates A are promoted to participate in the enzymatic reaction as much as possible, the enzymatic reaction can be continuously kept in the forward direction, the enzymatic reaction is more complete, more solid particle substrates A are converted into chiral beta-amino acid derivatives, and the yield of the chiral beta-amino acid derivatives is higher.
In a word, the preparation method of the chiral beta-amino acid derivative disclosed by the invention can be used for more effectively overcoming the defect that a substrate is dissolved in an organic solvent and then is added dropwise, solving the defect that the substrate is easy to agglomerate and separate out in the enzymatic reaction process, and promoting the yield of the chiral beta-amino acid derivative to be higher. Moreover, the preparation method provided by the invention is simpler and more convenient, the raw materials are easier to obtain, the repeatability is better, and the method has better industrial production prospect.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (14)
1. The preparation method of the chiral beta-amino acid derivative is characterized in that the chiral beta-amino acid derivative has a structure shown in the following general formula I:
in said general formula I, the stereogenic center marked with x has the (R) -or (S) -configuration; an enantiomeric excess of at least 70% compared to the corresponding enantiomer;
z is NR 2 R 3 ；NR 2 R 3 Is of the structure ofR 4 Is hydrogen or C which is unsubstituted or substituted by 1 to 5 fluorine atoms 1～4 An alkyl group;
r is aryl-C 1～2 Alkyl, wherein said aryl-C 1～2 The phenyl of the alkyl group is substituted with 1-3 substituents of fluorine;
wherein, the preparation method further comprises the following steps:
s1, mixing a first solvent, a biological enzyme catalyst and an amino donor to form a mixed raw material solution; the first solvent is a mixed solvent of water and an alcohol solvent, and the alcohol solvent is methanol; the biological enzyme catalyst is aminotransferase, and the aminotransferase is omega-aminotransferase; the amino donor being C 1～6 An alkylamine;
s2, adding a solid particle substrate A in a continuous solid strand form into the mixed raw material solution through a continuous feeding device to perform enzymatic reaction, wherein the structural formula of the solid particle substrate A is as followsR and Z have the same meanings as described above, giving the chiral β -amino acid derivative.
2. The method for producing chiral β -amino acid derivatives according to claim 1, characterized in that the addition rate of the solid particulate substrate a is 0.5-12.0 g/s.
3. The method for producing chiral β -amino acid derivatives according to claim 2, characterized in that the continuous solid flow is controlled to be multi-stranded during the addition of the solid particle substrate a, and the strands are independent from each other.
4. A process for the preparation of chiral β -amino acid derivatives as claimed in claim 3 wherein during the addition of said solid particulate substrate a, the plurality of said continuous solid streams are controlled to assume a swinging state relative to the liquid surface upon entry into said mixed feed solution.
5. The method for preparing chiral β -amino acid derivatives according to claim 2, characterized in that the addition time of the solid particle substrate a is 4-12 h.
6. The method for producing chiral β -amino acid derivatives according to claim 5, characterized in that the addition time of the solid particle substrate a is 6 to 10 hours.
7. The method for producing chiral β -amino acid derivatives according to claim 1, characterized in that the amount of the solid particulate substrate a added is 80-500 g per liter of the first solvent.
8. The process for the preparation of chiral β -amino acid derivatives according to any one of claims 1 to 7, characterized in that the molar ratio of the solid particulate substrate a and the amino donor is 1 (3-10).
9. The method for producing chiral β -amino acid derivatives according to any one of claims 1 to 7, characterized in that the mixed raw material solution further comprises a buffer salt system.
10. The method for preparing chiral β -amino acid derivatives according to claim 9, characterized in that the buffer salt system is one or more of potassium dihydrogen phosphate/dipotassium hydrogen phosphate, tris/hydrochloric acid, triethanolamine, barbital sodium/hydrochloric acid, glycine/sodium hydroxide or amino donor/hydrochloric acid.
11. The method for producing chiral β -amino acid derivatives according to claim 8, wherein the mass fraction of the alcohol solvent is 20-40%.
12. The method for producing a chiral β -amino acid derivative according to any one of claims 1 to 7, characterized in that said mixed raw material solution further comprises an amino mediator selected from one or more of pyridoxal phosphate, pyridoxamine phosphate or pyridoxine phosphate.
13. The process for preparing a chiral β -amino acid derivative according to any one of claims 1 to 7, characterized in that the reaction temperature is 20 to 60 ℃, the reaction pH is 7.0 to 11, and the reaction time is 18 to 36 hours during the enzymatic reaction.
14. The process for preparing a chiral β -amino acid derivative according to claim 13,
in the enzymatic reaction process, the reaction temperature is 35-55 ℃, and the reaction pH value is 8.0-10.5.
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