CN116024283A - Method for biocatalytic methylation of dextromethorphan precursor - Google Patents
Method for biocatalytic methylation of dextromethorphan precursor Download PDFInfo
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- CN116024283A CN116024283A CN202211603392.1A CN202211603392A CN116024283A CN 116024283 A CN116024283 A CN 116024283A CN 202211603392 A CN202211603392 A CN 202211603392A CN 116024283 A CN116024283 A CN 116024283A
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- dextromethorphan
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- MKXZASYAUGDDCJ-SZMVWBNQSA-N LSM-2525 Chemical compound C1CCC[C@H]2[C@@]3([H])N(C)CC[C@]21C1=CC(OC)=CC=C1C3 MKXZASYAUGDDCJ-SZMVWBNQSA-N 0.000 title claims abstract description 47
- 229960001985 dextromethorphan Drugs 0.000 title claims abstract description 46
- 239000002243 precursor Substances 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 32
- 230000011987 methylation Effects 0.000 title claims abstract description 22
- 238000007069 methylation reaction Methods 0.000 title claims abstract description 22
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- 238000006243 chemical reaction Methods 0.000 claims abstract description 26
- MEFKEPWMEQBLKI-AIRLBKTGSA-N S-adenosyl-L-methioninate Chemical class O[C@@H]1[C@H](O)[C@@H](C[S+](CC[C@H](N)C([O-])=O)C)O[C@H]1N1C2=NC=NC(N)=C2N=C1 MEFKEPWMEQBLKI-AIRLBKTGSA-N 0.000 claims abstract description 21
- 239000003054 catalyst Substances 0.000 claims abstract description 13
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 12
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 10
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- 239000011973 solid acid Substances 0.000 claims description 10
- 229910018072 Al 2 O 3 Inorganic materials 0.000 claims description 8
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 8
- 229910004298 SiO 2 Inorganic materials 0.000 claims description 8
- 239000003960 organic solvent Substances 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
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- 125000004970 halomethyl group Chemical group 0.000 claims description 4
- GZUXJHMPEANEGY-UHFFFAOYSA-N bromomethane Chemical compound BrC GZUXJHMPEANEGY-UHFFFAOYSA-N 0.000 claims description 3
- 238000011065 in-situ storage Methods 0.000 claims description 3
- ZRNSSRODJSSVEJ-UHFFFAOYSA-N 2-methylpentacosane Chemical compound CCCCCCCCCCCCCCCCCCCCCCCC(C)C ZRNSSRODJSSVEJ-UHFFFAOYSA-N 0.000 claims description 2
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- GEIAQOFPUVMAGM-UHFFFAOYSA-N ZrO Inorganic materials [Zr]=O GEIAQOFPUVMAGM-UHFFFAOYSA-N 0.000 claims description 2
- NEHMKBQYUWJMIP-NJFSPNSNSA-N chloro(114C)methane Chemical compound [14CH3]Cl NEHMKBQYUWJMIP-NJFSPNSNSA-N 0.000 claims description 2
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims description 2
- 125000005842 heteroatom Chemical group 0.000 claims description 2
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- NBVXSUQYWXRMNV-UHFFFAOYSA-N monofluoromethane Natural products FC NBVXSUQYWXRMNV-UHFFFAOYSA-N 0.000 claims description 2
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- 206010011224 Cough Diseases 0.000 description 5
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- 238000006555 catalytic reaction Methods 0.000 description 4
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 4
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
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- ZJUKTBDSGOFHSH-WFMPWKQPSA-N S-Adenosylhomocysteine Chemical compound O[C@@H]1[C@H](O)[C@@H](CSCC[C@H](N)C(O)=O)O[C@H]1N1C2=NC=NC(N)=C2N=C1 ZJUKTBDSGOFHSH-WFMPWKQPSA-N 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
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- OROGSEYTTFOCAN-DNJOTXNNSA-N codeine Chemical compound C([C@H]1[C@H](N(CC[C@@]112)C)C3)=C[C@H](O)[C@@H]1OC1=C2C3=CC=C1OC OROGSEYTTFOCAN-DNJOTXNNSA-N 0.000 description 2
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- BQJCRHHNABKAKU-KBQPJGBKSA-N morphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4O BQJCRHHNABKAKU-KBQPJGBKSA-N 0.000 description 2
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- 239000011780 sodium chloride Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- ILNSWVUXAPSPEH-USXIJHARSA-N 3-methoxy-morphinan Chemical compound C1CCC[C@H]2[C@H]3CC4=CC=C(OC)C=C4[C@]21CCN3 ILNSWVUXAPSPEH-USXIJHARSA-N 0.000 description 1
- 206010006458 Bronchitis chronic Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- 206010012335 Dependence Diseases 0.000 description 1
- LCGLNKUTAGEVQW-UHFFFAOYSA-N Dimethyl ether Chemical compound COC LCGLNKUTAGEVQW-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- JAQUASYNZVUNQP-USXIJHARSA-N Levorphanol Chemical compound C1C2=CC=C(O)C=C2[C@]23CCN(C)[C@H]1[C@@H]2CCCC3 JAQUASYNZVUNQP-USXIJHARSA-N 0.000 description 1
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- 206010046306 Upper respiratory tract infection Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000001350 alkyl halides Chemical class 0.000 description 1
- 230000029936 alkylation Effects 0.000 description 1
- 238000005804 alkylation reaction Methods 0.000 description 1
- 229940124584 antitussives Drugs 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
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- 229960004126 codeine Drugs 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- ZZVUWRFHKOJYTH-UHFFFAOYSA-N diphenhydramine Chemical compound C=1C=CC=CC=1C(OCCN(C)C)C1=CC=CC=C1 ZZVUWRFHKOJYTH-UHFFFAOYSA-N 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
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- 208000017574 dry cough Diseases 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- OROGSEYTTFOCAN-UHFFFAOYSA-N hydrocodone Natural products C1C(N(CCC234)C)C2C=CC(O)C3OC2=C4C1=CC=C2OC OROGSEYTTFOCAN-UHFFFAOYSA-N 0.000 description 1
- 208000030603 inherited susceptibility to asthma Diseases 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 229960003406 levorphanol Drugs 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000001767 medulla oblongata Anatomy 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 229940102396 methyl bromide Drugs 0.000 description 1
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- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 229920001983 poloxamer Polymers 0.000 description 1
- KCXFHTAICRTXLI-UHFFFAOYSA-N propane-1-sulfonic acid Chemical compound CCCS(O)(=O)=O KCXFHTAICRTXLI-UHFFFAOYSA-N 0.000 description 1
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- 239000011949 solid catalyst Substances 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 239000012130 whole-cell lysate Substances 0.000 description 1
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/584—Recycling of catalysts
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for biocatalytic dextromethorphan precursor methylation, which comprises the following steps: step one, preparing equipment; step two, synthesizing; in the first step, equipment required to be used in the synthesis process is prepared; wherein in the second step, S-adenosylmethionine analogue is taken as a methyl donor, and the reaction is carried out for 24-72 hours at the temperature of 10-40 ℃ in a solution, and a biocatalyst is used for catalyzing the methylation of the dextromethorphan precursor to obtain a corresponding product, wherein the ratio of the dosage of the S-adenosylmethionine analogue to the dosage of the dextromethorphan precursor is 1:0.1-10, and the ratio of the dosage of the catalyst to the dosage of the dextromethorphan precursor is 1:0.1-10; the method takes S-adenosylmethionine analogue as a methyl donor, and the methylation of dextromethorphan precursors is catalyzed by enzyme to obtain the corresponding product dextromethorphan; the method has the advantages of mild reaction conditions, environmental friendliness, simplicity in operation, wide raw material application range, high catalytic conversion rate and good chemical and regional selectivity.
Description
Technical Field
The invention relates to the technical field of biocatalytic conversion, in particular to a method for biocatalytic dextromethorphan precursor methylation.
Background
Dextromethorphan is a central antitussive, and mainly inhibits and delays the cough central of brain, and is dextroisomer of morphine levorphanol methyl ether, and can inhibit and delay cough central of medulla oblongata to exert central antitussive effect. The cough strength of the dextromethorphan Sha Fenzhen is equal to or slightly stronger than that of codeine, the pain relieving effect is not generated, the tolerance and addiction are not generated after long-term application, the respiration is not inhibited by the treatment dose, the oral absorption is good, the effect is realized within 15-30 minutes, the effect can be maintained for 3-6 hours, and the concentration of the proto-drug in the blood plasma is very low. The main active metabolite 3-methoxymorphinan of dextromethorphan has high concentration in plasma, t1/2 is 5 hours, is mainly used for dry cough, is suitable for common cold, acute or chronic bronchitis, bronchial asthma, sphagitis, tuberculosis and other cough caused by upper respiratory tract infection, and the synthesis steps are shown in figure 7, and the traditional chemical hydrocarbylating method (G.L. Pluronic, et. Molecular, 2002,7,697-705; U.S. Pat. No. 1973027200) generally reacts with haloalkane under the catalysis of strong alkali, the reaction condition is harsh, three wastes are more, and the environmental pollution is also great. The enzymatic catalysis condition is mild, the environment is friendly, the yield is high, and the development of a new method for catalyzing alkylation by the enzymatic catalysis provides a wider development space for the synthesis and application of dextromethorphan derivatives.
The biological enzyme catalysis has the outstanding advantages of mild reaction conditions, no pollution and three wastes, strong selectivity, green chemistry and the like, and is a main method for developing and replacing the traditional organic synthesis. The invention takes coenzyme S-adenosylmethionine (SAM) as a methylation donor, carries out methylation modification on dextromethorphan precursor, and releases an equivalent by-product S-adenosylhomocysteine (SAH).
The invention takes SAM food as hydrocarbylating donor to catalyze the hydrocarbylating of dextromethorphan precursor, and can realize the recycling of SAM analogues by adding solid acid catalyst. The enzyme-catalyzed dextromethorphan precursor has the advantages of mild hydrocarbylation reaction conditions, simple operation, wide raw material application range, environmental protection, no pollution, cyclic regeneration of hydrocarbylation donors, high catalytic conversion rate and chemical and regioselectivity.
Disclosure of Invention
The invention aims to provide a method for biocatalytic methylation of dextromethorphan precursors, which aims to solve the problems in the background art.
In order to achieve the above purpose, the present invention provides the following technical solutions: a method of biocatalytic dextromethorphan precursor methylation comprising the steps of: step one, preparing equipment; step two, synthesizing;
in the first step, equipment required to be used in the synthesis process is prepared;
in the second step, S-adenosylmethionine analogue is taken as a methyl donor, and the reaction is carried out for 24-72 hours at the temperature of 10-40 ℃ in a solution, and the biocatalyst is used for catalyzing the methylation of the dextromethorphan precursor to obtain a corresponding product, wherein the ratio of the dosage of the S-adenosylmethionine analogue to the dosage of the dextromethorphan precursor is 1:0.1-10, and the ratio of the dosage of the catalyst to the dosage of the dextromethorphan precursor is 1:0.1-10.
Preferably, in the second step, the expression is alkylated to methyl.
Preferably, in the second step, the heteroatom of the dextromethorphan precursor is N, and the cyclic regeneration of the methyl donor is methyl fluoride, methyl chloride, methyl bromide and methyl iodide.
Preferably, in the second step, the pH of the solution is 5.5-8.5.
Preferably, in the second step, the S-adenosylmethionine analogue can be regenerated in situ by adding a solid acid catalyst and a halogenated methyl group into a reaction system; the solid acid catalyst is SiO 2 ,Al 2 O 3 ,ZrO 2 ,SiO 2 -Al 2 O 3 ,SiO 2 -MgO,SiO 2 -ZrO 2 ,B 2 O 3 -Al 2 O 3 ,Cr 2 O 3 -Al 2 O 3 Natural zeolite, znCl 2 ,AlCl 3 ,MnSO 4 ,CuSO 4, The dosage of the solid acid catalyst is 0.1-3 times of the dosage of S-adenosylmethionine analogue substances. The amount of halomethyl groups is 1-10 times the amount of S-adenosylmethionine analog material.
Preferably, in the second step, when the solid acid catalyst and the halomethyl group are added to the reaction system, an organic solvent is required to be added simultaneously to dissolve the halomethyl group, and the organic solvent is one or more of hexane, pentane, petroleum ether, methanol, ethanol, isopropanol, dimethyl sulfoxide (DMSO), acetone, N-Dimethylformamide (DMF).
Compared with the prior art, the invention has the beneficial effects that: the method for biologically catalyzing the methylation of the dextromethorphan precursor takes SAM food as a hydrocarbylating donor to catalyze the hydrocarbylating of the dextromethorphan precursor, and can realize the recycling of SAM analogues by adding a solid acid catalyst. The enzyme-catalyzed dextromethorphan precursor has the advantages of mild hydrocarbylation reaction conditions, simple operation, wide raw material application range, environmental protection, no pollution, cyclic regeneration of hydrocarbylation donors, high catalytic conversion rate and chemical and regioselectivity.
Drawings
FIG. 1 is a schematic diagram of the reaction synthesis in example 1 of the present invention;
FIG. 2 is a flow chart of the method of the present invention;
FIG. 3 shows a hydrogen nuclear magnetic resonance spectrum of the target product in example 1 of the present invention;
FIG. 4 is a nuclear magnetic resonance chart of the target product in example 1 of the present invention;
FIG. 5 is a high resolution mass spectrum of the target product in example 1 of the present invention;
FIG. 6 is a schematic diagram of a specific method in embodiment 3 of the present invention;
FIG. 7 is a diagram of a prior art method of synthesizing steps.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Referring to fig. 1-6, an embodiment of the present invention is provided:
example 1:
a method of biocatalytic dextromethorphan precursor methylation comprising the steps of: step one, preparing equipment; step two, synthesizing;
in the first step, equipment required to be used in the synthesis process is prepared;
wherein in the second step above, purified COMT (final concentration 6.3 mg/mL), me-SAM (final concentration 4.0 mM), dextromethorphan precursor (final concentration 4.0 mM) was added to 30mL of a solution containing 50mM Tris Cl, pH8.0,1mM MgCl 2 Adding 30mL of NaCl with mass concentration of 26% into 0.5mM EDTA solution for reaction at 40 ℃ for 2 hours, stopping reaction, centrifuging to remove precipitate, extracting supernatant with Ethyl Acetate (EA) for three times, removing organic solvent by rotary evaporation, separating and purifying crude product by silica gel column chromatography, and eluting with Ethyl Acetate (EA)/Petroleum Ether (PE) =1:3 (v/v) to obtain target product dextromethorphan with yield of 99.5%; meanwhile, the target product is as follows: dextromethorphan nuclear magnetic resonance hydrogen spectrum: 1 HNMR(400MHz,CDCl 3 )δ=7.06-7.04(d,J=8.0,1H),6.82(s,1H),6.73-6.71(d,J=8.0,1H),3.80(s,3H),3.15-3.06(m,2H),2.76-2.58(m,3H),2.37-2.30(m,1H),2.26(s,3H),1.77-1.74(m,1H),1.66-1.51(m,3H),1.40-1.26(m,3H);
dextromethorphan nuclear magnetic resonance carbon spectrum: 13 CNMR(101MHz,CDCl3)δ=158.3,141.9,130.2,128.4,111.3,110.7,55.3,51.3,46.9,43.7,42.9,39.4,38.4,37.2,33.827.0,26.9,22.3;
right America Sha Fengao resolved mass spectrum, HRMS (ESI) M/z [ M+H ] +calcualatedfor C18H26NO+:272.1890; found 272.1886.
Example 2:
a method of biocatalytic dextromethorphan precursor methylation comprising the steps of: step one, preparing equipment; step two, synthesizing;
in the first step, equipment required to be used in the synthesis process is prepared;
wherein in the second step, the COMT-expressing E.coli recombinant strain whole cell lysate (final concentration 5.7 mg/mL), me-SAM (final concentration 4.0 mM), dextromethorphan precursor (final concentration 4.0 mM) was mixed with 30mL of a solution containing 50mM Tris Cl, pH8.3,1mM Dithiothreitol (DTT), 20mM MgCl 2 In a solution of 5mM EDTA, reacting for 4 hours at 40 ℃, adding 30mL of NaCl with mass concentration of 26% to terminate the reaction, centrifuging to remove precipitate, extracting supernatant with Ethyl Acetate (EA) for three times, removing organic solvent by rotary evaporation, separating and purifying a crude product by silica gel column chromatography, and obtaining a target product dextromethorphan with a yield of 98.3% by using Ethyl Acetate (EA)/Petroleum Ether (PE) =1:3 (v/v) as an eluent.
Example 3:
a method of biocatalytic dextromethorphan precursor methylation comprising the steps of: step one, preparing equipment; step two, synthesizing;
in the first step, equipment required to be used in the synthesis process is prepared;
wherein in the second step, hamster kidney cell lysate (final concentration 7.5 mg/mL), me-SAM (final concentration 4.0 mM), dextromethorphan precursor (final concentration 4.0 mM) was mixed with 25mL of 50mM3- (N-morpholino) propanesulfonic acid (MOPS), pH6.4,1mM Dithiothreitol (DTT), 10mM MgCl 2 In a solution of 10mM EDTA, the reaction is carried out for 6 hours at 30 ℃,30 mL of 1MHCl is added to terminate the reaction, the solution is centrifuged to remove the precipitate, the supernatant is extracted with Ethyl Acetate (EA) for three times, the organic solvent is removed by rotary evaporation, the crude product is separated and purified by silica gel column chromatography, the Ethyl Acetate (EA)/Petroleum Ether (PE) =1:3 (v/v) is taken as an eluent, the target product dextromethorphan is obtained, the yield is 95.9%, and the solid catalyst and CH3X are utilized in the step, and the SAM analogue is recycled in situ, wherein the specific method principle is shown in figure 6.
Based on the above, the invention has the advantages of mild reaction conditions, good selectivity, renewable hydrocarbylated donors, high catalytic conversion rate and almost quantitative completion; the high catalytic conversion rate is reflected in that the hydrocarbylation reaction of the dextromethorphan precursor can be completely converted by adjusting the dosage of the biocatalyst, prolonging the reaction time and using a SAM analogue recycling strategy.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Claims (6)
1. A method of biocatalytic dextromethorphan precursor methylation comprising the steps of: step one, preparing equipment; step two, synthesizing; the method is characterized in that:
in the first step, equipment required to be used in the synthesis process is prepared;
in the second step, S-adenosylmethionine analogue is taken as a methyl donor, and the reaction is carried out for 24-72 hours at the temperature of 10-40 ℃ in a solution, and the biocatalyst is used for catalyzing the methylation of the dextromethorphan precursor to obtain a corresponding product, wherein the ratio of the dosage of the S-adenosylmethionine analogue to the dosage of the dextromethorphan precursor is 1:0.1-10, and the ratio of the dosage of the catalyst to the dosage of the dextromethorphan precursor is 1:0.1-10.
2. A method of biocatalytic dextromethorphan precursor methylation according to claim 1, wherein: in the second step, the expression is alkylated to methyl.
3. A method of biocatalytic dextromethorphan precursor methylation according to claim 1, wherein: in the second step, the hetero atom of the dextromethorphan precursor is N, and the methyl donor is recycled to be methyl fluoride, methyl chloride, bromomethane and methyl iodide.
4. A method of biocatalytic dextromethorphan precursor methylation according to claim 1, wherein: in the second step, the pH value of the solution is 5.5-8.5.
5. A method of biocatalytic dextromethorphan precursor methylation according to claim 1, wherein: in the second step, the S-adenosylmethionine analogue can be regenerated in situ by adding a solid acid catalyst and halogenated methyl into a reaction system; the solid acid catalyst is SiO 2 ,Al 2 O 3 ,ZrO 2 ,SiO 2 -Al 2 O 3 ,SiO 2 -MgO,SiO 2 -ZrO 2 ,B 2 O 3 -Al 2 O 3 ,Cr 2 O 3 -Al 2 O 3 Natural zeolite, znCl 2 ,AlCl 3 ,MnSO 4 ,CuSO 4, The dosage of the solid acid catalyst is 0.1-3 times of the dosage of S-adenosylmethionine analogue substances. The amount of halomethyl groups is 1-10 times the amount of S-adenosylmethionine analog material.
6. A method of biocatalytic dextromethorphan precursor methylation according to claim 1, wherein: in the second step, when the solid acid catalyst and the halogenated methyl are added into the reaction system, an organic solvent is required to be added simultaneously to dissolve the halogenated methyl, wherein the organic solvent is one or a mixture of more than two of hexane, pentane, petroleum ether, methanol, ethanol, isopropanol, dimethyl sulfoxide (DMSO), acetone and N, N-Dimethylformamide (DMF).
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