CN105734089A - An asymmetric synthesis method for (R)-3-amino piperidine derivatives - Google Patents
An asymmetric synthesis method for (R)-3-amino piperidine derivatives Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 46
- PEUGKEHLRUVPAN-RXMQYKEDSA-N (3r)-piperidin-3-amine Chemical class N[C@@H]1CCCNC1 PEUGKEHLRUVPAN-RXMQYKEDSA-N 0.000 title claims abstract description 19
- 238000011914 asymmetric synthesis Methods 0.000 title claims description 5
- 238000006243 chemical reaction Methods 0.000 claims abstract description 60
- 102000003929 Transaminases Human genes 0.000 claims abstract description 25
- 108090000340 Transaminases Proteins 0.000 claims abstract description 25
- 239000003054 catalyst Substances 0.000 claims abstract description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 17
- 102000004190 Enzymes Human genes 0.000 claims description 14
- 108090000790 Enzymes Proteins 0.000 claims description 14
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical compound CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 claims description 13
- 235000007682 pyridoxal 5'-phosphate Nutrition 0.000 claims description 12
- 239000011589 pyridoxal 5'-phosphate Substances 0.000 claims description 12
- 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 12
- 229960001327 pyridoxal phosphate Drugs 0.000 claims description 12
- 239000000284 extract Substances 0.000 claims description 11
- XUWHAWMETYGRKB-UHFFFAOYSA-N piperidin-2-one Chemical class O=C1CCCCN1 XUWHAWMETYGRKB-UHFFFAOYSA-N 0.000 claims description 10
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 6
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 claims description 6
- 101100246550 Caenorhabditis elegans pyr-1 gene Proteins 0.000 claims description 4
- 241000186359 Mycobacterium Species 0.000 claims description 4
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 claims description 4
- 229940097267 cobaltous chloride Drugs 0.000 claims description 4
- 238000001212 derivatisation Methods 0.000 claims description 4
- 230000003100 immobilizing effect Effects 0.000 claims description 4
- 239000000376 reactant Substances 0.000 claims description 4
- -1 trichloro-ethoxycarbonyl Chemical group 0.000 claims description 4
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 claims description 3
- LJCZNYWLQZZIOS-UHFFFAOYSA-N 2,2,2-trichlorethoxycarbonyl chloride Chemical compound ClC(=O)OCC(Cl)(Cl)Cl LJCZNYWLQZZIOS-UHFFFAOYSA-N 0.000 claims description 3
- 125000005519 fluorenylmethyloxycarbonyl group Chemical group 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 3
- 125000005931 tert-butyloxycarbonyl group Chemical group [H]C([H])([H])C(OC(*)=O)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 3
- 241000142559 Mycobacterium vanbaalenii Species 0.000 claims description 2
- 239000007864 aqueous solution Substances 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims description 2
- 229920001429 chelating resin Polymers 0.000 claims description 2
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- 150000002500 ions Chemical class 0.000 claims description 2
- 239000002773 nucleotide Substances 0.000 claims description 2
- 125000003729 nucleotide group Chemical group 0.000 claims description 2
- 235000006408 oxalic acid Nutrition 0.000 claims description 2
- 229920000647 polyepoxide Polymers 0.000 claims description 2
- 238000000746 purification Methods 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 230000003287 optical effect Effects 0.000 abstract description 4
- 239000002904 solvent Substances 0.000 abstract description 4
- 238000005576 amination reaction Methods 0.000 abstract description 2
- 230000007613 environmental effect Effects 0.000 abstract 1
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- 239000000047 product Substances 0.000 description 18
- 239000007788 liquid Substances 0.000 description 9
- 238000005160 1H NMR spectroscopy Methods 0.000 description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 8
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 8
- 239000000706 filtrate Substances 0.000 description 7
- 239000012064 sodium phosphate buffer Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000000605 extraction Methods 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- KWLMDGDJYYHBAL-UHFFFAOYSA-K trisodium ethanol phosphate Chemical compound [Na+].[Na+].[Na+].CCO.[O-]P([O-])([O-])=O KWLMDGDJYYHBAL-UHFFFAOYSA-K 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 238000010926 purge Methods 0.000 description 4
- GGPNYXIOFZLNKW-ZJIMSODOSA-N (3r)-piperidin-3-amine;dihydrochloride Chemical compound Cl.Cl.N[C@@H]1CCCNC1 GGPNYXIOFZLNKW-ZJIMSODOSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000009903 catalytic hydrogenation reaction Methods 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- CUYKNJBYIJFRCU-UHFFFAOYSA-N 3-aminopyridine Chemical compound NC1=CC=CN=C1 CUYKNJBYIJFRCU-UHFFFAOYSA-N 0.000 description 2
- 108010093096 Immobilized Enzymes Proteins 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000005194 fractionation Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- DYHSDKLCOJIUFX-UHFFFAOYSA-N tert-butoxycarbonyl anhydride Chemical compound CC(C)(C)OC(=O)OC(=O)OC(C)(C)C DYHSDKLCOJIUFX-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- NIPYQLPZPLBOLF-UHFFFAOYSA-N 3'-hydroxy-6'-[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyspiro[2-benzofuran-3,9'-xanthene]-1-one Chemical compound OC1C(O)C(O)C(CO)OC1OC1=CC=C2C3(C4=CC=CC=C4C(=O)O3)C3=CC=C(O)C=C3OC2=C1 NIPYQLPZPLBOLF-UHFFFAOYSA-N 0.000 description 1
- 0 C*([C@](CCC1)C*1C(O)=O)C(O)=O Chemical compound C*([C@](CCC1)C*1C(O)=O)C(O)=O 0.000 description 1
- 108010067722 Dipeptidyl Peptidase 4 Proteins 0.000 description 1
- 102100025012 Dipeptidyl peptidase 4 Human genes 0.000 description 1
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- VZTDIZULWFCMLS-UHFFFAOYSA-N ammonium formate Chemical compound [NH4+].[O-]C=O VZTDIZULWFCMLS-UHFFFAOYSA-N 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000006264 debenzylation reaction Methods 0.000 description 1
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
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- 238000005984 hydrogenation reaction Methods 0.000 description 1
- FUKUFMFMCZIRNT-UHFFFAOYSA-N hydron;methanol;chloride Chemical class Cl.OC FUKUFMFMCZIRNT-UHFFFAOYSA-N 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
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- 238000005891 transamination reaction Methods 0.000 description 1
- LEAHFJQFYSDGGP-UHFFFAOYSA-K trisodium;dihydrogen phosphate;hydrogen phosphate Chemical compound [Na+].[Na+].[Na+].OP(O)([O-])=O.OP([O-])([O-])=O LEAHFJQFYSDGGP-UHFFFAOYSA-K 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method of performing asymmetric amination by utilizing a transaminase catalyst to prepare (R)-3-amino piperidine derivatives. Compared with other preparing methods, the method has the characteristics of a higher product concentration, higher optical purity of a product, easily recovered solvents, mild reaction conditions, capability of being environmental friendly, simple and convenient operation and easy industrial scaling-up.
Description
Technical field
The invention belongs to technical field of bioengineering, be specifically related to the method that one utilizes transaminase-catalyzed dose of asymmetric synthesis (R)-3-amino piperidine derivatives.
Background technology
(R)-3-amino piperidine derivatives is the important intermediate in many pharmaceutical synthesis processes.Such as, (R)-3-amino piperidine is the important intermediate of synthesis Egelieting or BI 1356, and this kind of medicine is DPP-IV inhibitor, can be used for treating type ii diabetes.(R) chemical constitution of-3-amino piperidine, Egelieting and BI 1356 is as follows respectively:
In PCT Patent application WO2007075630 and WO2008028654, with 3-aminopyridine for initiation material, through catalytic hydrogenation, fractionation with become salt to obtain (R)-3-amino piperidine dihydrochloride, reaction equation following (not including salt-forming steps):
The method route is succinct, but the catalytic hydrogenation of pyridine to use catalyst costly and higher pressure, commercial production inconvenience, and split process wastes the isomer of half, make production cost increase at double, and cause the discharge of discarded isomer, contaminated environment.
Adv.Synth.Catal.2008; 350 (6); 807-812 reports with nicotiamide for raw material, protect with de-Boc by catalytic hydrogenation, amino Boc protection, Hoffman degraded, fractionation, become salt five step to react, obtain (R)-3-amino piperidine.The method needs also exist for splitting isomer, thus wasting the isomer of half, not only makes production cost increase at double, and causes the discharge of discarded isomer, contaminated environment.
Disclosed in PCT Patent application WO2008102720, racemic N-benzyl-3-piperidine formamide is split by enzyme and obtains (R)-N-benzyl-3-piperidine formamide, reset through Hoffman, hydrogenation debenzylation, one-tenth salt obtain (R)-3-amino piperidine dihydrochloride, reaction equation following (not including salt-forming steps):
This route creates the discarded isomer of half equally, and environment is unfriendly.
Summary of the invention
Prepare the problems such as yield is low, cost of material is expensive, reaction is incomplete, corresponding selection is not high in (R)-3-amino piperidine derivatives technique for the asymmetric amination reported, it is an object of the invention to provide the method that one utilizes enzyme catalysis that 3-derivative of piperidone is converted into (R)-3-amino piperidine derivatives.
To achieve these goals, the present invention adopts the following technical scheme that.
A kind of method of asymmetric synthesis (R)-3-amino piperidine derivatives, it is characterised in that: 3-derivative of piperidone enzymatically turns aminating reaction, obtains (R)-3-amino piperidine derivatives.
In said method, catalysis 3-derivative of piperidone carries out the enzyme catalyst of asymmetric ammoxidation formation (R)-3-amino piperidine derivatives and is derived from the transaminase of mycobacterium (Mycobacteriumvanbaalenii) PYR-1, or the culture of its recombinant conversion body, it is also possible to it is the transformant cell by obtaining after this culture centrifugation or with the goods of its processing.Here, " goods of processing " refer to the extract obtained by transformant, or the separation product by the transaminase in extract being easily separated and/or purification obtains, or the immobilizing product obtained by immobilization transformant cell or extract or the separating product of transformant.
In the present invention, described in derive from the transaminase of mycobacterium PYR-1 or restructuring transaminase is documented in the Chinese patent application that application number is 201410169882.4, nucleotide sequence coded shown in SEQIDNo.1 therein.At this, the full text of this application is introduced in the lump using as reference.
In the present invention, above-mentioned immobilizing product is by restructuring transaminase is fixed on a kind of enzyme immobilization carrier the immobilization transaminase obtained.Described enzyme immobilization carrier can be that epoxy resin performs the derivatization obtain through imido oxalic acid (IDA) and cobaltous chloride successively, it is also possible to is that ion chelating resin performs the derivatization through cobaltous chloride and obtains.
Above-mentioned immobilization transaminase and preparation method thereof is documented in the Chinese patent application that application number is 201410262154.8 in detail, is introduced in the lump by the full text of this application at this using as reference.
In the method for the present invention, described 3-derivative of piperidone has structure as shown below:
Wherein, R is selected from H, tertbutyloxycarbonyl (Boc), fluorenylmethyloxycarbonyl (Fmoc), allyloxycarbonyl, trichloro-ethoxycarbonyl (Troc), benzyloxycarbonyl group (Cbz), the tert-butyl group.
Correspondingly, the structure of (the R)-3-amino piperidine derivatives prepared is as follows:
In method disclosed by the invention, described in turn the reaction condition of aminating reaction and can select by the normal condition of this type of reaction of this area.
Preferably, described method comprises the steps: in the ethanol water that pH value is 7.0-10.0, under the existence of 2-aminopropane. and optional pyridoxal 5-phosphate (PLP), under the effect of above-mentioned enzyme catalyst (such as transaminase, restructuring transaminase or immobilization transaminase), 3-derivative of piperidone carries out the asymmetric aminating reaction that turns, and forms (R)-3-amino piperidine derivatives.
In said method, 3-derivative of piperidone concentration in reactant liquor is preferably 1-800mmol/L.Catalyst is catalytically effective amount, it is preferred to 0.1-50g/L.The consumption of 2-aminopropane. is preferably 1-60g/L.The consumption of PLP is preferably 0-8.0mmol/L.Described aqueous solution is this area conventional buffers, as long as its pH scope is at 7.0-10.0, it is preferable that phosphate buffer, such as phosphoric acid-sodium phosphate buffer.The concentration of phosphate buffer is preferably 0.05-0.1mol/L, and described concentration refers to the total concentration of conjugate acid and base in buffer solution.Concentration of alcohol is preferably 5%-50% (V/V).Described asymmetric transamination reaction carries out preferably under vibration or stirring condition, and reaction temperature is preferably 20-55 DEG C, and the response time was as the criterion with the time that production concentration in course of reaction is no longer constantly improve.Asymmetric turn after aminating reaction terminates, Chiral Amine product can be extracted from reactant liquor by this area conventional method.
Under the premise without prejudice to essence of the present invention and this area general knowledge, above-mentioned each optimum condition can combination in any, obtain the preferred embodiments of the invention.
Agents useful for same of the present invention and raw material are all commercially.
The actively progressive effect of the present invention is in that: for it have been reported that reaction yield is low, the problem such as cost of material is expensive, reaction is incomplete, corresponding selection is not high, the invention discloses one utilization specifically transaminase-catalyzed dose carry out asymmetric amidized method.When the substrate of catalytic level up to 0.8mol/L, the optical purity of product is still up to more than 99%.Relative to other preparation method existing, using the present invention to prepare production concentration and the optical purity height of gained, solvent easily reclaims, and reaction condition is gentle, environmentally friendly, easy and simple to handle, it is easy to industry is amplified, and has good prospects for commercial application.
Detailed description of the invention
Further illustrate the present invention below in conjunction with embodiment, but should not be construed as limiting the scope of the invention.In the following example, for unreceipted actual conditions, it was shown that be the normal condition according to this area or manufacturer it is proposed that condition carry out.
The method recorded in the Chinese patent application utilizing application number to be 201410169882.4 prepares restructuring transaminase and mutant thereof, the immobilization transaminase that the method recorded in the Chinese patent application utilizing application number to be 201410262154.8 prepares, carries out turning aminating reaction.
Embodiment 1
At 100ml ethanol-sodium phosphate buffer, (ethanol volumetric concentration is 50%, pH8.5) 591mg 2-aminopropane. is added in, being subsequently adding the immobilization transaminase extremely final protein concentration prepared according to the method recorded in Chinese patent application 201410262154.8 is 1g/L, add 1.99g3-oxo-piperidine-1-t-butyl formate and 24.7mg pyridoxal 5-phosphate, insert bottom at 50 DEG C with syringe needle and carry out nitrogen purging.In course of reaction with HPLC measure conversion ratio to 99% or reaction 24 hours after stopped reaction.Reaction is filtered to remove immobilized enzyme after terminating, and filtrate adjusts pH to 10.5-11 with NaOH, heats to 60 DEG C and stirs 30min removing 2-aminopropane., being subsequently adding the Bis(tert-butoxycarbonyl)oxide (Boc of final concentration of 120mmol/L2O), room temperature reaction is overnight.Extracting by 100mL ethyl acetate, extracting twice, combining extraction liquid, add anhydrous sodium sulfate and dry, filter, filtrate is distilled, and obtains (R)-3-t-butoxycarbonyl amino piperidines-1-t-butyl formate 2.85g.After measured, conversion ratio is 95%, and ee value is 99%.
Product structure is as follows:
Said structure is confirmed by 1HNMR.
1HNMR(300MHz,CDCl3):δ8.0(s,1H),3.43-3.68(d,2H),3.60(m,1H),3.32-3.35(t,2H),1.60-1.85(dt,2H),1.45-1.55(m,2H),1.40(s,18H).
In embodiment, HPLC testing conditions: developing solvent is ethyl acetate: methanol=3:1, developing solvent adds 1 2-aminopropane..Iodine cylinder, 1,2,3-indantrione monohydrate all can develop the color.
Conversion ratio assay method is: reactant liquor dilution in acetonitrile 100 times, take 20 μ L after centrifugal on Agilent1200HPLC, analyze conversion ratio, analytical column is AgilentEclipseXAD-C18 reverse phase silica gel post, mobile phase is water: acetonitrile=40:60, separately adding 10mmol/L ammonium formate, flow velocity is 1mL per minute, and detection wavelength is 210nm and 260nm, the retention time turning ammonia product is 4.6min, and the retention time of substrate is 7.4min.
The ee values determination method turning ammonia product is: detect the optical purity e.e value of product on Agilent1200HPLC, DaicelChiralpakAD-H chiral chromatographic column (4.6 × 150mm) is adopted to be analyzed, mobile phase is ethanol: normal heptane: diethylamine: water=60:40:0.1:0.1, flow velocity is 1mL per minute, column temperature is 35 DEG C, retention time is respectively as follows: substrate 5.6min, (S)-product: 7.9min；(R)-product: 9.7min.Detection wavelength is 210nm.
Embodiment 2
At 200ml ethanol-sodium phosphate buffer, (ethanol volumetric concentration is 50%, pH8.5) 2.6g 2-aminopropane. is added in, being subsequently adding the immobilization transaminase extremely final protein concentration prepared according to the method recorded in Chinese patent application 201410262154.8 is 5g/L, add 8.0g3-oxo-piperidine-1-benzyl formate and 84.8mg pyridoxal 5-phosphate, insert bottom at 50 DEG C with syringe needle and carry out nitrogen purging.In course of reaction measure conversion ratio to 99% or reaction 24 hours after stopped reaction.Reaction is filtered to remove immobilized enzyme after terminating, and filtrate adjusts pH to 10.5-11 with NaOH, heats to 60 DEG C and stirs 30min removing 2-aminopropane., being subsequently adding the Bis(tert-butoxycarbonyl)oxide (Boc of final concentration of 120mmol/L2O), room temperature reaction 2 hours, extract by 100mL ethyl acetate, extracting twice, combining extraction liquid, add anhydrous sodium sulfate and dry, filter, filtrate is distilled, and obtains (R)-3-t-butoxycarbonyl amino piperidines-1-benzyl formate 10.2g.Measuring conversion ratio according to the method for embodiment 1 is 89%, and ee value is 99%.
Product structure is as follows:
Said structure is confirmed by 1HNMR.
1HNMR(300MHz,CDCl3):δ8.0(s,1H),7.19(s,5H),5.34(s,2H),3.43-3.68(d,2H),3.60(m,1H),3.32-3.35(t,2H),1.60-1.85(dt,2H),1.45-1.55(m,2H),1.40(s,18H).
Embodiment 3
At 100ml ethanol-sodium phosphate buffer, (ethanol volumetric concentration is 50%, pH8.5) 2-aminopropane. 0.1g is added in, immobilization transaminase prepared by addition method described in Chinese patent application 201410262154.8 is 0.1g/L to final protein concentration, add 0.17g3-piperidones and 0.29mg pyridoxal 5-phosphate, react under 50 DEG C of magnetic agitation.With HPLC mensuration conversion ratio to 99% stopped reaction in course of reaction.Reaction is filtered with gauze, the washing with alcohol of filtering residue 10mL80% three times after terminating, and filters, filtrate merges, and extracts by 50mL ethyl acetate, extracting twice, combining extraction liquid, adds anhydrous sodium sulfate and is dried overnight, and obtains (R)-3-amino piperidine 0.16g.Measuring conversion ratio according to the method for embodiment 1 is 92%, and ee value is 97%.
Product structure is as follows:
Embodiment 4
At 100ml ethanol-sodium phosphate buffer, (ethanol volumetric concentration is 50%, pH8.5) 2-aminopropane. 5.7g is added in, the crude enzyme liquid of transaminase's mutant that addition is prepared according to the method recorded in Chinese patent application 201410169882.4 to final protein concentration is 30g/L, add 21.8g3-oxo-piperidine-1-formic acid-2,2,2-trichloro ethyl ester and 178mg pyridoxal 5-phosphate, insert bottom at 50 DEG C with syringe needle and carry out nitrogen purging.In course of reaction measure conversion ratio to 99% or reaction 24 hours after stopped reaction.Reaction extracts by 100mL ethyl acetate, extracting twice, combining extraction liquid, adds anhydrous sodium sulfate and dry after terminating, and filters, and filtrate is distilled, and obtains (R)-3-amino piperidine-1-formic acid-2,2,2-trichloro ethyl ester 21.2g.Measuring conversion ratio according to the method for embodiment 1 is 97%, and ee value is 98%.
Product structure is as follows:
Said structure is confirmed by 1HNMR.
1HNMR(300MHz,CDCl3):δ4.84(s,2H),3.43-3.68(d,2H),3.32-3.35(t,2H),2.63(t,1H),2.0(s,2H),1.60-1.85(dt,2H),1.45-1.55(m,2H).
Embodiment 5
At 100ml ethanol-sodium phosphate buffer, (ethanol volumetric concentration is 50%, pH8.5) 0.1g 2-aminopropane. is added in, being subsequently added into the crude enzyme liquid of transaminase's mutant to the final protein concentration prepared according to the method recorded in Chinese patent application 201410169882.4 is 0.1g/L, add 0.31g3-oxo-piperidine-1-allyl formate and 3.2mg pyridoxal 5-phosphate, insert bottom at 50 DEG C with syringe needle and carry out nitrogen purging.In course of reaction measure conversion ratio to 99% or reaction 24 hours after stopped reaction.Reaction extracts by 100mL ethyl acetate, extracting twice, combining extraction liquid, adds anhydrous sodium sulfate and dry after terminating, and filters, and filtrate is distilled, and obtains (R)-3-amino piperidine-1-allyl formate 0.30g.Measuring conversion ratio according to the method for embodiment 1 is 98%, and ee value is 98%.
Product structure is as follows:
Said structure is confirmed by 1HNMR.
1HNMR(300MHz,CDCl3):δ5.89(s,1H),5.22-5.23(s,2H),4.75(s,2H),3.43-3.68(d,2H),3.32-3.35(t,2H),2.63(t,1H),2.0(s,2H),1.60-1.85(dt,2H), 1.45-1.55(m,2H).
Embodiment 6
At 1L ethanol-sodium phosphate buffer, (ethanol volumetric concentration is 50%, pH8.5) 20.0g 2-aminopropane. is added in, the crude enzyme liquid of transaminase's mutant that addition is prepared according to the method recorded in Chinese patent application 201410169882.4 to final protein concentration is 30g/L, add 33.5g3-piperidones and 600mg pyridoxal 5-phosphate, react under 50 DEG C of magnetic agitation.With HPLC mensuration conversion ratio to 99% stopped reaction in course of reaction.Reaction extracts by 50mL ethyl acetate, extracting twice, combining extraction liquid, adds anhydrous sodium sulfate and be dried overnight, obtain (R)-3-amino piperidine 30.5g after terminating.Measuring conversion ratio according to the method for embodiment 1 is 90%, and ee value is 97%.
Product structure is as follows:
Embodiment 7
Take (R)-3-t-butoxycarbonyl amino piperidines-1-t-butyl formate 30g that method shown in embodiment 1 prepares, it is dissolved in 20mL dichloromethane, add saturated HCl-methanol solution (saturated concentration 10-11M) 30mL, it is stirred at room temperature 1 hour, precipitation filters, and by washed with dichloromethane, obtain (R)-3-amino piperidine dihydrochloride 16.4g, yield 95%.
Product structure is as follows:
Claims (12)
1. the method for asymmetric synthesis (R)-3-amino piperidine derivatives, it is characterised in that: 3-derivative of piperidone enzymatically turns aminating reaction, obtains (R)-3-amino piperidine derivatives.
2. method according to claim 1, it is characterized in that: the enzyme catalyst of use is derived from the transaminase of mycobacterium (Mycobacteriumvanbaalenii) PYR-1, or the culture of its recombinant conversion body, or by by the transformant cell obtained after this culture centrifugation or with the goods of its processing.
3. method according to claim 2, it is characterized in that: described in derive from the transaminase of mycobacterium PYR-1 or restructuring transaminase is documented in the Chinese patent application that application number is 201410169882.4, nucleotide sequence coded shown in SEQIDNo.1 therein.
4. method according to claim 2, it is characterized in that: the goods of described processing refer to the extract obtained by transformant, or the separation product by the transaminase in extract being easily separated and/or purification obtains, or the immobilizing product obtained by immobilization transformant cell or extract or the separating product of transformant.
5. method according to claim 4, it is characterised in that: described immobilizing product is by restructuring transaminase is fixed on a kind of enzyme immobilization carrier the immobilization transaminase obtained.
6. method according to claim 5, it is characterized in that: described enzyme immobilization carrier is that epoxy resin performs the derivatization obtain through imido oxalic acid (IDA) and cobaltous chloride successively, or ion chelating resin performs the derivatization through cobaltous chloride and obtains.
7. method according to claim 1, it is characterised in that: described 3-derivative of piperidone is structured with:
Wherein, R is selected from H, tertbutyloxycarbonyl (Boc), fluorenylmethyloxycarbonyl (Fmoc), allyloxycarbonyl, trichloro-ethoxycarbonyl (Troc), benzyloxycarbonyl group (Cbz), the tert-butyl group.
8. method according to claim 1, it is characterised in that: the structure of described (R)-3-amino piperidine derivatives is as follows:
Wherein, R is selected from H, tertbutyloxycarbonyl (Boc), fluorenylmethyloxycarbonyl (Fmoc), allyloxycarbonyl, trichloro-ethoxycarbonyl (Troc), benzyloxycarbonyl group (Cbz), the tert-butyl group.
9. the method according to any one of claim 1-8, it is characterized in that, comprise the steps: in the ethanol water that pH value is 7.0-10.0, under the existence of 2-aminopropane. and optional pyridoxal 5-phosphate (PLP), under the effect of enzyme catalyst, 3-derivative of piperidone carries out the asymmetric aminating reaction that turns, and forms (R)-3-amino piperidine derivatives.
10. method according to claim 9, it is characterised in that: 3-derivative of piperidone concentration in reactant liquor is 1-800mmol/L, and enzyme catalyst concentration is 0.1-50g/L, and the concentration of 2-aminopropane. is 1-60g/L, and the concentration of pyridoxal 5-phosphate is 0-8.0mmol/L.
11. method according to claim 9, it is characterised in that: described aqueous solution is phosphate buffer, and concentration of alcohol is 5%-50% (V/V).
12. method according to claim 9, it is characterised in that: the described asymmetric reaction temperature turning aminating reaction is 20-55 DEG C.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105974036A (en) * | 2016-07-31 | 2016-09-28 | 合肥远志医药科技开发有限公司 | Detection method of aminopyridine isomer |
| CN106520719A (en) * | 2016-11-25 | 2017-03-22 | 华东理工大学 | S-shaped omega-transaminase ATA-W12 as well as gene and application thereof |
| CN107604020A (en) * | 2017-09-27 | 2018-01-19 | 上海合全药物研发有限公司 | The method that living things catalysis prepares the amino piperidine of (S) 1 N benzene methoxycarbonyl group 3 |
| CN110724675A (en) * | 2019-10-31 | 2020-01-24 | 宁波酶赛生物工程有限公司 | Transaminase catalyst and method for synthesizing (R) -1-tert-butoxycarbonyl-3-aminopiperidine by enzyme method |
| CN114107275A (en) * | 2021-11-19 | 2022-03-01 | 辽宁凯莱英医药化学有限公司 | Enzyme immobilization carrier and preparation method thereof, immobilized enzyme and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103189519A (en) * | 2010-07-14 | 2013-07-03 | 帝斯曼知识产权资产管理有限公司 | (R)-selective amination |
| CN103865964A (en) * | 2014-03-14 | 2014-06-18 | 上海朴颐化学科技有限公司 | Method for synthesizing (R)-3-amino-piperidine by adopting transaminase method |
| CN104178536A (en) * | 2014-07-31 | 2014-12-03 | 洛阳华荣生物技术有限公司 | Biological preparation method for R-3-aminopiperidine |
-
2014
- 2014-12-11 CN CN201410758416.XA patent/CN105734089A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103189519A (en) * | 2010-07-14 | 2013-07-03 | 帝斯曼知识产权资产管理有限公司 | (R)-selective amination |
| CN103865964A (en) * | 2014-03-14 | 2014-06-18 | 上海朴颐化学科技有限公司 | Method for synthesizing (R)-3-amino-piperidine by adopting transaminase method |
| CN104178536A (en) * | 2014-07-31 | 2014-12-03 | 洛阳华荣生物技术有限公司 | Biological preparation method for R-3-aminopiperidine |
Non-Patent Citations (3)
| Title |
|---|
| 刘婷婷: "药物中间体( Ｒ)-3-氨基哌啶二( 三氟乙酸) 盐的合成", 《南昌大学学报( 工科版)》 * |
| 史作清: "《吸附分离树脂在医药工业中的应用》", 30 September 2008, 化学工业出版社 * |
| 王金胜: "《酶工程》", 30 September 2007, 中国农业出版社 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105974036A (en) * | 2016-07-31 | 2016-09-28 | 合肥远志医药科技开发有限公司 | Detection method of aminopyridine isomer |
| CN106520719A (en) * | 2016-11-25 | 2017-03-22 | 华东理工大学 | S-shaped omega-transaminase ATA-W12 as well as gene and application thereof |
| CN106520719B (en) * | 2016-11-25 | 2019-06-07 | 华东理工大学 | A kind of ω of S type-transaminase ATA-W12 and its gene and application |
| CN107604020A (en) * | 2017-09-27 | 2018-01-19 | 上海合全药物研发有限公司 | The method that living things catalysis prepares the amino piperidine of (S) 1 N benzene methoxycarbonyl group 3 |
| CN110724675A (en) * | 2019-10-31 | 2020-01-24 | 宁波酶赛生物工程有限公司 | Transaminase catalyst and method for synthesizing (R) -1-tert-butoxycarbonyl-3-aminopiperidine by enzyme method |
| CN114107275A (en) * | 2021-11-19 | 2022-03-01 | 辽宁凯莱英医药化学有限公司 | Enzyme immobilization carrier and preparation method thereof, immobilized enzyme and preparation method thereof |
| CN114107275B (en) * | 2021-11-19 | 2024-05-17 | 辽宁凯莱英医药化学有限公司 | Enzyme immobilization carrier and preparation method thereof, immobilized enzyme and preparation method thereof |
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