A kind of asymmetric heterocyclic ketone Aldol catalysts and its reaction method
Technical field
The invention belongs to chemical catalyst field, and in particular to catalytic applications of the papain in organic synthesis,
More particularly to application of the papain as asymmetric heterocyclic ketone Aldol catalysts, and using papain
The method of enzymatic asymmetry heterocyclic ketone Aldol reactions.
Background technology
The structure of C-C keys is the basis of organic synthesis.Aldol reactions are considered as one of important method for building C-C.Cause
This controls the relative and absolute configuration of Aldol reaction products to be just particularly important in synthesis.Organic molecule and organic gold
The asymmetric Aldol reaction of category catalysis is largely reported.In these reports, the acceptor of Aldol reactions has been obtained for
Research well;However, the research to the donor scope of application is but very limited.Xiao Wenjing etc. (Chen J.R., LiX.Y.,
XingX.N., XiaoW.J., Sterically and Electronically Tunable and
BifunctionalOrganocatalysts:Design and Application in Asymmetric Aldol
Reaction of Cyclic Ketones with Aldehydes[J].The Journal of Organic
Chemistry, 2006, (71):8198-8202.) and (Pihko P.M., the Laurikainen K.M., Usano such as Kaavi
A., Nyberg A.I., KaaviJ.A., Effect of additives on the proline-catalyzed ketone-
Aldehyde aldol reactions [J] .Tetrahedron, 2006, (62):317.) the asymmetric heterocyclic ketone of report
Aldol reactions achieve good effect.But their catalyst has a deficiency, for example catalyst synthesis is cumbersome, to ring
Border is harmful to, non-renewable and poisonous etc..And living things catalysis is due to gentle and potential renewable with high selectivity, reaction condition
Property, therefore widely it is considered a kind of effective and green modern organic synthesis means.2008, its grade of remaining filial piety was reported first
Lipase-catalyzed asymmetric Aldol reaction.(the Guan Z., Jian-PingFu, Yan-HongHe, lipase- such as He Yanhong
catalysed asymmetric aldol reaction of heterocyclic ketones with aldehydes
[J] .Tetrahedron Letters, 2012, (53):4959-4961.) report enzymatic heterocyclic ketone asymmetry
Aldol is reacted, and efficient but more and tool spatial selectivity catalysis potential enzyme is urgently exploited.
Papain (Papain) is a kind of new phytoenzyme up for utilization, is a kind of half important Guang
Serine protease, for Peptide systhesis.Chinese invention patent CN 201110063369.3 discloses chymopapain conduct
The application of direct asymmetric aldol reaction catalyst, aromatic aldehyde or heteroaryl aldehyde and dialkyl group are used as using chymopapain
The application of direct asymmetric aldol reaction catalyst between ketone or cyclanone.But change the heterocyclic ketone of multi-functional catalysis with
The direct asymmetric aldol reaction of aldehyde is not reported also.
The content of the invention
In view of this, the technical problem to be solved in the present invention is to provide a kind of asymmetric heterocyclic ketone Aldol reactions
Catalyst.Another the to be solved technical problem of the present invention is to provide a kind of catalyst of asymmetric heterocyclic ketone Aldol reactions
Reaction method.
The present invention solves above-mentioned technical problem by following technical proposals:
A kind of catalyst of asymmetric heterocyclic ketone Aldol reactions, it is characterised in that contain papain.
A kind of reaction method of the catalyst of asymmetric heterocyclic ketone Aldol reactions, its step include:
(1) added into reaction vessel on phenyl ring and be connected with the aromatic aldehyde and heterocyclic ketone of electron-withdrawing substituent, aromatic aldehyde with it is miscellaneous
The mol ratio of cyclic ketones is 1: (0.5-5), then solubilizer, and papain enzyme catalyst, papain 25-
200mg/ (0.25mmol heterocyclic ketones);
(2) 15-40 DEG C of controlling reaction temperature, stirring reaction 12-144 hours;
(3) reacting liquid filtering, filter cake are washed with extractant, merging filtrate and cleaning solution, then adds deionized water dilution, used
Extractant extracts;
(4) by after the extract anhydrous sodium sulfate drying of extraction, then carry out vacuum distillation and remove solvent, produce product.
In order to obtain superior technique effect, heterocyclic ketone described in step (1) is in piperidones, pyranone, thiapyrones
One or more combination;In step (1), the aromatic aldehyde that electron-withdrawing substituent is connected with the phenyl ring is 4- fluorobenzaldehydes, 3- fluorine
Benzaldehyde, 2- fluorobenzaldehydes, 4- chlorobenzaldehydes, 3- chlorobenzaldehydes, 2- chlorobenzaldehydes, 4- bromobenzaldehydes, 3- bromobenzaldehydes, 2- bromines
Benzaldehyde, 4- nitrobenzaldehydes, 3- nitrobenzaldehydes, 2- nitrobenzaldehydes, 4- cyanobenzaldehydes, 3- cyanobenzaldehydes, 2- cyanogen
One or more combinations in benzaldehyde;Organic solvent in step (1) is tetrahydrofuran, DMF, second
One or more combinations in nitrile, dimethyl sulfoxide, isopropanol, Isosorbide-5-Nitrae-dioxane, chloroform;In step (1), the solvent is to go
The mixed solvent of ionized water and organic solvent composition, V (H2O)/V(H2O+ organic solvents) volume ratio is (0-3)/10;Solvent is
(0.5-5) mL/ (0.25mmol heterocyclic ketones);In step (1), the mol ratio of aromatic aldehyde and heterocyclic ketone is preferably 1: 0.5, papaya
Protease is preferably 100mg/ (0.25mmol heterocyclic ketones), and solvent is preferably 1mL/ (0.25mmol heterocyclic ketone), solvent for go from
The mixed solvent of sub- water and organic solvent composition, V (H2O)/V(H2O+ organic solvents)=2.0: 10;In step (2), reaction temperature
Preferably 35 DEG C, the reaction time is preferably 120 hours;In step (3), the extractant is dichloromethane or ethyl acetate;Step
Suddenly after (4), the product with the mixed solution that flash column chromatography eluant, eluent is that volume ratio is petroleum ether and ethyl acetate,
Obtain 37% yield.
Present invention firstly discovers that the catalyst that papain can react as asymmetric heterocyclic ketone Aldol, especially
The asymmetric heterocyclic ketone Aldol reactions applied between aromatic aldehyde or heteroaryl aldehyde and heterocyclic ketone, have that catalytic activity is good, mapping
The advantages that selectivity is high, economic, environmentally friendly, has not only widened the application of papain, and be asymmetric heterocyclic ketone
Aldol reactions provide a kind of excellent biological enzyme agent, are had broad application prospects in organic synthesis field.
Chemical reagent used in the present invention is as follows:
Papain (Papain, >=3unit/mg, article No.:P104243), the brilliant pure biochemical technology share in Shanghai is limited
The purchase of company of company;
Piperidones;
Pyranone;
Thiapyrones;
Tetrahydrofuran;
DMF;
Acetonitrile;
Dimethyl sulfoxide;
Isopropanol;
Isosorbide-5-Nitrae-dioxane;
Chloroform;
Deionized water;
The aromatic aldehyde of electron-withdrawing substituent is connected with phenyl ring, is 4- fluorobenzaldehydes, 3- fluorobenzaldehydes, 2- fluorobenzaldehydes, 4-
Chlorobenzaldehyde, 3- chlorobenzaldehydes, 2- chlorobenzaldehydes, 4- bromobenzaldehydes, 3- bromobenzaldehydes, 2- bromobenzaldehydes, 4- nitrobenzaldehydes,
3- nitrobenzaldehydes, 2- nitrobenzaldehydes, 4- cyanobenzaldehydes, 3- cyanobenzaldehydes, one kind in 2- cyanobenzaldehydes or more
Kind combination;
Reaction kit:The double A multiplex vavuum pump of circulating water type of SHB-III;RE52CS-1 type Rotary Evaporators;ABL-224 types
Assay balance;DLSB-5/10 type low-temperature cooling fluid circulating pumps;DF-101S type heat collecting type constant-temperature heating magnetic stirring apparatus;DHJF-
4002 type cryogenic thermostat stirring reactive baths;DHG-9076A type electric heating constant-temperature blowing drying boxes;KQ250DE type ultrasonic cleaners.
Analytical instrument:Thin-layered chromatography (GF254 silica gel plates) monitors reaction process;ZF-2I type ultraviolet analysis instrument for three purposed;It hurry up
Fast column chromatography uses 100-200 mesh silica gel pressured columns;Using Bruker AV-300 type nmr determination hydrogen nuclear magnetic resonances
Spectrum (1H NMR) and carbon spectrum (13C NMR).Using the enantiomeric excess value of HPLC chiral fixed phase method measure product
(ee) and diastereomeric excess value (dr), chiral stationary phase use AD-H, OD-H, AS-H and OJ-H chiral column.Given in embodiment
The yield gone out is product yield after purification, and anti represents trans product, and syn represents cis-product.
Brief description of the drawings
Fig. 1 is the Aldol reaction time progress curves of papain enzymatic.
Embodiment
Comparative example
A kind of reaction method, its step include:
(1) added into reaction vessel on phenyl ring and be connected with the aromatic aldehyde and heterocyclic ketone of electron-withdrawing substituent, aromatic aldehyde with it is miscellaneous
The mol ratio of cyclic ketones is 1: (0.5-5), adds organic solvent and the mixed solvent 1mL of deionized water, volume ratio is V (H2O)/
V(H2O+ organic solvents)=(0-3)/10;
(2) in temperature control (15-40 DEG C) stirring reaction (12-144 hours);
(3) reacting liquid filtering, filter cake are washed with extractant (dichloromethane), merging filtrate and cleaning solution, then add go from
Sub- water dilution, is extracted with extractant;
(4) by after the extract anhydrous sodium sulfate drying of extraction, then carry out vacuum distillation and remove solvent, do not detect product.
Embodiment 1
A kind of catalyst of asymmetric heterocyclic ketone Aldol reactions, it is characterised in that contain papain.
Embodiment 2:The influence of solvent
The reaction method of the catalyst of asymmetric heterocyclic ketone Aldol reactions, its step include described in embodiment 1:
(1) 0.50mmol 4- nitrobenzaldehydes and 0.25mmol N- tertbutyloxycarbonyls -4- are added into reaction vessel
Piperidones, the mol ratio of 4- nitrobenzaldehydes and N- tertbutyloxycarbonyl -4- piperidones is 1: 0.5, adds 1mL organic solvent
With the mixed solvent of deionized water composition, the wherein volume ratio of deionized water and mixed solvent is 1: 10, and papain is
50mg;
(2) 25 DEG C of controlling reaction temperature, stirring reaction 120 hours;
(3) reacting liquid filtering, filter cake are washed with extractant dichloromethane, merging filtrate and cleaning solution, then adds deionization
Water dilutes, and is extracted with extractant dichloromethane;
(4) by after the extract anhydrous sodium sulfate drying of extraction, then carry out vacuum distillation and remove solvent, produce product.
Reaction equation is:
The screening of the Aldol reaction dissolvents of the papain enzymatic of table 1
Reaction media is to influence an important factor for reaction is carried out, therefore we have studied a series of solvents to reaction process
Act on (table 1).Relative to the advantages of enzymatic, enzyme equally shows wonderful property in non-aqueous system in aqueous medium,
Such as the stability of enzyme is improved, non-natural reaction etc. can be catalyzed.It can clearly be seen that reaction dissolvent to a kind wood from table
The catalytic activity of melon protease influences very big.We have found that enzyme is in hydrophilic solvent tetrahydrofuran, acetonitrile, dimethyl sulfoxide and N, N-
Preferable catalytic activity is shown in dimethylformamide.
In addition, in order to prove that papain has catalytic action to Aldol reactions, we devise a series of controls
Experiment.As it can be seen from table 1 in the case of not enzyme-added, Aldol reactions can hardly be carried out (table 1, sequence number 9).However,
When bovine serum albumin (B.S.A.) and the sodium acid carbonate catalysis of non-enzymatic, 44% and 45% raceme product is respectively obtained, has been demonstrate,proved
Bright non-zymoprotein can be catalyzed Aldol reactions to a certain extent, but not have stereoselectivity, and ee values are 0.With Urea treatment mistake
Ficin enzymic catalytic reaction still obtained 5% product, and there is no stereoselectivity.This illustrates the three of enzyme simultaneously
It is essential to the stereoselectivity of product to tie up structure.We are also sub- with serpin methyl first sulfidomethyl
Sulfone (MMTS) treatment enzyme, the product (table 1, sequence number 11) of trace is as a result only observed on chromatographic sheet, this eliminates
The possibility of the amino acid residue catalytic of enzyme, and the avtive spot of ficin is further demonstrated to being catalyzed more work(
Energy property serves decisive action.Above experimental result has absolutely proved that the asymmetric direct asymmetric Aldol reaction must occur without flower
On the special construction in fruit proteolytic catalytic activity site.
Embodiment 3:The influence of water content
The reaction method of the catalyst of asymmetric heterocyclic ketone Aldol reactions, its step include described in embodiment 1:
(1) 0.50mmol 4- nitrobenzaldehydes and 0.25mmol N- tertbutyloxycarbonyls -4- are added into reaction vessel
Piperidones, the mol ratio of 4- nitrobenzaldehydes and N- tertbutyloxycarbonyl -4- piperidones is 1: 0.5, adds the organic molten of 0.5mL
Agent CH3CN and deionized water mixed solvent, wherein H2O/(H2O+CH3CN volume ratio) is 0-3/10, and papain is
50mg;
(2) 25 DEG C of controlling reaction temperature, stirring reaction 120 hours;
(3) reacting liquid filtering, filter cake are washed with extractant dichloromethane, merging filtrate and cleaning solution, then adds deionization
Water dilutes, and is extracted with extractant dichloromethane;
(4) by after the extract anhydrous sodium sulfate drying of extraction, then carry out vacuum distillation and remove solvent, produce product.
Water is typically considered to play a part of lubricant in the structure of enzyme, and it can increase the toughness of enzymatic structure, system
In suitable water can improve the stability and catalytic activity of enzyme, accordingly, it is determined that the optimum content of reaction system reclaimed water is very heavy
Will.We devise experiment to find the optimum moisture content of Aldol reactions.We allow reaction in different proportion [H2O/(H2O+
CH3CN carry out in mixed reaction solvent)], be as a result listed in Table 2.From table it will be seen that with water content from 0 increase
0.20 is added to, yield and ee values are continuously increased (table 2, sequence number 1-5).When water content is 0.20, yield and ee values reach optimal, this
When yield be 34%.However, once water content, more than 0.20, yield and ee values start to reduce.Therefore, water content of the present invention is preferred
0.15-0.25, more preferably 0.2.
The influence that the water content of table 2 is reacted papain enzymatic Aldol
Embodiment 4:The influence of substrate rate of charge
The reaction method of the catalyst of asymmetric heterocyclic ketone Aldol reactions, its step include described in embodiment 1:
(1) 0.125-1.25mmol 4- nitrobenzaldehydes and 0.25mmol N- tertiary butyloxycarbonyls are added into reaction vessel
Base -4- piperidones, the mol ratio of 4- nitrobenzaldehydes and N- tertbutyloxycarbonyl -4- piperidones is 0.5-5: 1, adds 2mL's
Organic solvent CH3CN and deionized water mixed solvent, wherein H2O/(H2O+CH3CN volume ratio) is 2/10, papain
Enzyme is 50mg;
(2) 25 DEG C of controlling reaction temperature, stirring reaction 120 hours;
(3) reacting liquid filtering, filter cake are washed with extractant dichloromethane, merging filtrate and cleaning solution, then adds deionization
Water dilutes, and is extracted with extractant dichloromethane;
(4) by after the extract anhydrous sodium sulfate drying of extraction, then carry out vacuum distillation and remove solvent, produce product.
When the ratio of 4- nitrobenzaldehydes and ketone is 3: 1, the preferable ee values of highest yield (table 3, sequence number 4) have been obtained,
But yield is also more or less the same when rate of charge is 2: 1, ee value highests, dr values are good when being also 3: 1 compared with rate of charge, from atom economy
From the point of view of property and reaction result, substrate rate of charge of the invention (mol ratio of aldehyde/ketone) is preferably 1-3, and more preferably 2.
The influence that the rate of charge of table 3 is reacted papain enzymatic Aldol.
Embodiment 5:The influence of enzyme amount
The reaction method of the catalyst of asymmetric heterocyclic ketone Aldol reactions, its step include described in embodiment 1:
(1) 0.5mmol 4- nitrobenzaldehydes and 0.25mmol N- tertbutyloxycarbonyl -4- piperazines are added into reaction vessel
Pyridine ketone, the mol ratio of 4- nitrobenzaldehydes and N- tertbutyloxycarbonyl -4- piperidones is 2: 1, adds 5mL organic solvent
CH3CN and deionized water mixed solvent, wherein H2O/(H2O+CH3CN volume ratio) is 2/10, papain 25-
200mg;
(2) 25 DEG C of controlling reaction temperature, stirring reaction 120 hours;
(3) reacting liquid filtering, filter cake are washed with extractant dichloromethane, merging filtrate and cleaning solution, then adds deionization
Water dilutes, and is extracted with extractant dichloromethane;
(4) by after the extract anhydrous sodium sulfate drying of extraction, then carry out vacuum distillation and remove solvent, produce product.
As can be seen from Table 4, when enzyme carrying capacity increases to 150mg from 25mg, yield persistently raises (table 4, sequence number 1-
5), but during 75mg and 150mg enzyme amount yield is more or less the same, and ee values and dr value highests during enzyme amount 75mg, therefore the present invention
Enzyme amount is preferably 75-150mg, more preferably 75mg.
The influence that the enzyme carrying capacity of table 4 is reacted papain enzymatic Aldol
Embodiment 6:The influence of reaction temperature
The reaction method of the catalyst of asymmetric heterocyclic ketone Aldol reactions, its step include described in embodiment 1:
(1) 0.5mmol 4- nitrobenzaldehydes and 0.25mmol N- tertbutyloxycarbonyl -4- piperazines are added into reaction vessel
Pyridine ketone, the mol ratio of 4- nitrobenzaldehydes and N- tertbutyloxycarbonyl -4- piperidones is 2: 1, adds 3mL organic solvent
CH3CN and deionized water mixed solvent, wherein H2O/(H2O+CH3CN volume ratio) is 2/10, and papain is
75mg;
(2) 15-45 DEG C of controlling reaction temperature, stirring reaction 120 hours;
(3) reacting liquid filtering, filter cake are washed with extractant dichloromethane, merging filtrate and cleaning solution, then adds deionization
Water dilutes, and is extracted with extractant dichloromethane;
(4) by after the extract anhydrous sodium sulfate drying of extraction, then carry out vacuum distillation and remove solvent, produce product.
Reaction temperature is also very big to the activity influence of enzyme, as being shown in table 5, when reaction temperature is 35 DEG C,
Yield highest (table 5, sequence number 1-6), and reaction temperature be 30 DEG C when ee values it is best.Therefore the reaction temperature of the present invention is preferably
30-40 DEG C, more preferably 30 DEG C.
The influence that the temperature of table 5 is reacted papain enzymatic Aldol
Embodiment 7:The influence in reaction time
The reaction method of the catalyst of asymmetric heterocyclic ketone Aldol reactions, its step include described in embodiment 1:
(1) 0.5mmol 4- nitrobenzaldehydes and 0.25mmol N- tertbutyloxycarbonyl -4- piperazines are added into reaction vessel
Pyridine ketone, the mol ratio of 4- nitrobenzaldehydes and N- tertbutyloxycarbonyl -4- piperidones is 2: 1, adds 4mL organic solvent
CH3CN and deionized water mixed solvent, wherein H2O/(H2O+CH3CN volume ratio) is 2/10, and papain is
100mg;
(2) 35 DEG C of controlling reaction temperature, stirring reaction 12-144 hours;
(3) reacting liquid filtering, filter cake are washed with extractant dichloromethane, merging filtrate and cleaning solution, then adds deionization
Water dilutes, and is extracted with extractant dichloromethane;
(4) by after the extract anhydrous sodium sulfate drying of extraction, then carry out vacuum distillation and remove solvent, produce product.
The present invention has investigated reaction process, and the time graph of whole process is as shown in Figure 1.Yield is held within first day to the 5th day
Continuous increase, after the 5th day, increase unobvious.Ee values are little with the change in reaction time, and the reaction time crosses and become with decline
Gesture.Therefore, the reaction time of the invention is preferably 120 hours.
Embodiment 8:Heterocyclic ketone and the versatility of aromatic aldehyde Aldol reactions are investigated
The reaction method of the catalyst of asymmetric heterocyclic ketone Aldol reactions, its step include described in embodiment 1:
(1) addition 0.5mmol aromatic aldehyde and 0.25mmol heterocyclic ketone into reaction vessel, aromatic aldehyde and heterocyclic ketone
Mol ratio is 2: 1, adds 1mL organic solvent CH3CN and deionized water mixed solvent, wherein H2O/(H2O+CH3CN)
Volume ratio is 2/10, papain 150mg;
(2) 35 DEG C of controlling reaction temperature, stirring reaction 120 hours;
(3) reacting liquid filtering, filter cake are washed with extractant dichloromethane, merging filtrate and cleaning solution, then adds deionization
Water dilutes, and is extracted with extractant dichloromethane;
(4) by after the extract anhydrous sodium sulfate drying of extraction, then carry out vacuum distillation and remove solvent, produce product.
The expansion situation of substrate is as shown in table 6.
Reaction equation is as follows:
The heterocyclic ketone of the papain enzymatic of table 6 and the expansion situation of aromatic aldehyde Aldol reaction substrates
As can be seen from Table 6, the heterocyclic ketone of papain enzymatic and the Aldol of aromatic aldehyde react to have obtained preferably
Yield (21-37%) (table 6, sequence number 1-9), it has been found that the steric hindrance and electronic effect pair of aromatic aldehyde benzene ring substituents
Reaction result has a significant impact.The product that 2- nitrobenzoyl aldehyde reactions with maximum steric hindrance obtain is with optimal ee values, still
Yield can not even obtain as the increase of steric hindrance reduces rapidly.Such as thiapyrones and piperidones and the 2- with maximum steric hindrance
Nitrobenzaldehyde does not react, and the reaction of pyranone and 2- nitrobenzaldehydes increases with phenyl ring steric hindrance, and yield declines and ee values
Improve (table 6, sequence number 7).In addition, all reacted when electron-withdrawing substituent is connected with aromatic aldehyde phenyl ring preferable.And aromatic aldehyde phenyl ring
On when being connected with electron donating group, have to trace product.Such case is probably because electron-withdrawing substituent adds carbonyl
The electropositive of carbon, it is easier to by the attack of negative electrical charge on the carbon of ketone α positions, thereby promote the progress of reaction.We do not obtain
The Aldol products of electron-donating group aromatic aldehyde.