CN109593804A - A kind of method of enzymatic rapid synthesis nitrobenzimidazole analog derivative - Google Patents
A kind of method of enzymatic rapid synthesis nitrobenzimidazole analog derivative Download PDFInfo
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Abstract
The invention discloses a kind of methods of enzymatic rapid synthesis nitrobenzimidazole analog derivative, the method are as follows: using the ratio between amount of substance for 1:1~5 6- nitrobenzimidazole and acrylics compound as raw material, using Lipozyme RM IM as catalyst, using DMSO solvent as reaction dissolvent, Lipozyme RM IM is uniformly filled in the reaction channel of microfluidic channel reactor, the reaction channel internal diameter of the microfluidic channel reactor is 0.8~2.4mm, a length of 0.5~1.0m of reaction channel；Raw material and reaction dissolvent is set continuously to be passed through progress Michael addition reaction in reaction channel, controlling Michael addition reaction temperature is 40~55 DEG C, the Michael addition reaction time is 25~40min, collects reaction solution online, and reaction solution obtains nitrobenzimidazole analog derivative through conventional post-processing.The present invention has the advantages that the reaction time is short, selectivity is high and yield is high.
Description
(1) technical field
The present invention relates to a kind of methods of enzymatic rapid synthesis nitrobenzimidazole analog derivative.
(2) background technique
Imidazoles is a kind of important 5-membered aromatic heterocycle, due to its unique rich electrical structure, shows excellent biology
Activity is widely used in the fields such as pesticide, medicine, artificial material, artificial receptors, supermolecule ligand and bionic catalyst.It is special
It is not in pharmaceutical chemistry, a large amount of mature glyoxaline compounds resist as anticancer drug, antimicrobial agents, anti-inflammatory drug
Histamine drug, anti-nerve drug and blood-pressure drug etc. clinically use, and have significant curative effect to treatment disease.
Michael addition is to form one of the important tool of C-C, C-N, C-O, C-X key, is that the classics in organic synthesis add
At reaction.Michael addition reaction is generally required and is carried out under strong acid or strong base catalyst, severe reaction conditions, will lead to environment dirt
Dye and energy waste.In order to find more efficient synthetic method, people have developed a series of metallic catalysts in recent years, such as
KF/Al2O3、Y(NO3)3·6H2O、CeCl3、Bi(OTf)3Deng, but these catalyst can generate environmentally harmful substance, together
When many side reactions of association, seriously reduce the selectivity of reaction and the yield of target product.Solid supported is used according to another report
Type catalyst, ionic liquid etc. are reacted, but these reactions are long there are the reaction time, catalyst system preparation process is complicated
The disadvantages of.Thus, the green syt new technology of Michael addition reaction is explored as the research hotspot in organic synthesis field.
Enzymic catalytic reaction due to it efficiently, green and specificity become one of effective tool of green chemical synthesis by force.Its
Excellent selectivity and mild reaction condition cause the broad interest of the fields scientist such as chemical, pharmacy, material.But
It is restriction of the enzymatic reaction there is solvent to substrate dissolution and solvent polarity to enzyme activity inhibition etc., the reaction time is often very long
(24-96h) is not very high to specific substrates conversion ratio, thus develops one kind based on micro-fluidic on the basis of traditional enzymatic reaction
The new synthesis technology of the enzymatic glyoxaline compound of technology becomes our goal in research.
In recent years, the continuous flow reactor within the scope of micron or millimeter with channel size was in organic synthesis
It is widely applied.Compared with conventional chemical reactor, one of micro-fluidic reactor the most significant advantage is using harmful reagent
When safety.This is from the higher specific surface area of microreactor and better thermally conductive and mass-transfer performance, hot-spot phenomenon
Obviously inhibited.The lesser scale of microreactor also prevents the accumulation of inside reactor dangerous substance simultaneously.Therefore, micro- anti-
It answers device technology to provide the unique method for executing ultrafast exothermic reaction, and allows to execute via highly unstable or even explosive
The reaction that intermediate carries out.
Up to the present, domestic that starting conceptual phase is in enzymatic Michael addition reaction, there is also it is certain not
Foot, and the biological method multiselect is catalyzed with acylase, it is expensive, longer reaction time (24-96h) is needed, and right
In specific substrates reaction conversion ratio be not especially desirable.In order to develop a kind of efficient green imidazole derivative synthesis it is new
Technology, we have studied the method for online synthesizing nitryl benzimidazoles derivative lipase-catalyzed in micro passage reaction, purports
In the new technology of the nitrobenzimidazole analog derivative for finding a kind of high-efficiency environment friendly synthesized online.
(3) summary of the invention
The technical problem to be solved in the present invention is to provide lipase-catalyzed online synthesis in a kind of microfluidic channel reactor
The new process of nitrobenzimidazole analog derivative has the advantages that the reaction time is short, yield is high.
In order to solve the above technical problems, the present invention adopts the following technical scheme:
A kind of method of lipase-catalyzed online synthesizing nitryl benzimidazoles derivative, the method is using micro-fluidic logical
Road reactor, the microfluidic channel reactor include syringe pump, syringe, reaction channel and product collector, the note
Emitter is installed in syringe pump, is connected to by first interface with the entrance of the reaction channel, and the product collector passes through the
The outlet of two interfaces and the reaction channel, the reaction channel internal diameter be 0.8~2.4mm, reaction channel a length of 0.5~
1.0m；The described method includes: the 6- nitrobenzimidazole with the ratio between amount of substance for 1:1~5 and acrylics shown in formula 1
Compound is raw material, using Lipozyme RM IM as catalyst, using dimethyl sulfoxide as reaction dissolvent, by the fat
Enzyme Lipozyme RM IM is uniformly filled in reaction channel, raw material and reaction dissolvent is placed in syringe, the syringe
Raw material and reaction dissolvent are continuously passed through in the reaction channel under the promotion of syringe pump and carry out Michael addition reaction,
Controlling reaction temperature is 40~55 DEG C, and the reaction time is 25~40min, collects reaction solution online by product collector, described
Reaction solution post-treated respectively obtain 5- nitro benzo shown in 6- nitrobenzimidazole analog derivative and formula 3 shown in formula 2
Imidazole derivative；The interior to greatest extent of institute's catalyst filling, the addition of the catalyst can be accommodated in the reaction channel
Amount is calculated as 0.025~0.05g/mL with the volume of the reaction dissolvent；In reaction system, acrylics shown in the formula 1
The concentration of compound is 0.1~0.5mmol/mL,
In formula 1,2 and formula 3, the R is CH2CH2CH2CH3Or C (CH3)3。
Further, in the microfluidic channel reactor that the present invention uses, the syringe number can be one or more,
Depending on specific reaction requirement.Reaction raw materials of the present invention are two kinds, it is preferable to use two syringes, specifically, the injection
Device is the first syringe and the second syringe respectively, and first connecting pipe is Y type or T-type pipeline, first note
Emitter and the second note syringe are connected to two interfaces of the Y type or T-type pipeline and by the Y type or T-types
Pipeline is connected with the reaction channel, is increased by reactant molecule contact and the collision probability of microchannel, is made two bursts of reactions
Liquid stream is mixed and is reacted in public reaction channel.
Further, more specifically, method of the present invention includes the following steps:
The method includes the following steps: 6- nitrobenzimidazole and 1 institute of formula with the ratio between amount of substance for 1:1~5
The acrylics compound shown is raw material, using 0.5~1.0g Lipozyme RM IM as catalyst, with dimethyl sulfoxide
For reaction dissolvent, the Lipozyme RM IM is uniformly filled in reaction channel, is first dissolved with dimethyl sulfoxide
6- nitrobenzimidazole is loaded in the first syringe, and the acrylics compound shown in dimethyl sulfoxide dissolution type 1 is loaded on the
In two syringes；First syringe, the second syringe are loaded in same syringe pump again, then in the syringe pump
It is synchronous push under summarize raw material and reaction dissolvent by the Y type or T-type pipeline after into carrying out in reaction channel
Michael addition reaction, control reaction temperature are 40~55 DEG C, and the reaction time is 25~40min, online by product collector
Reaction solution is collected, the reaction solution is post-treated to respectively obtain 3 institute of 6- nitrobenzimidazole analog derivative shown in formula 2 and formula
The 5- nitrobenzimidazole analog derivative shown；In reaction system, the concentration of acrylics compound shown in the formula 1 is
0.1~0.5mmol/mL.
Heretofore described first syringe is consistent with the specification of the second syringe, 6- described in first syringe
The concentration of nitrobenzimidazole is usually 0.1mmol/mL.
Further, the microfluidic channel reactor further includes insulating box, and the reaction channel is placed in insulating box,
Reaction temperature can be effectively controlled with this.The insulating box can require voluntarily to select according to reaction temperature, for example water-bath is permanent
Incubator etc..
The present invention is unlimited for the material of reaction channel, it is recommended to use green, the material of environmental protection, such as silicone tube；For
The shape of reaction channel is preferably curved shape, it is ensured that reaction solution stably passes through.
In the present invention, the Lipozyme RM IM believes the quotient of (novozymes) company production using Novi
Product, be it is a kind of prepared by microorganism, 1,3 position-specifics, food-grade lipase (EC 3.1.1.3) are on particle silica gel
Preparation.It is obtained from Rhizomucor miehei, it is micro- with a kind of gene-modified aspergillus oryzae (Aspergillus oryzae)
Biology is by submerged fermentation production.
Lipozyme RM IM is uniformly filled in reaction channel by the method for the present invention, can directly will by physical method
Granular catalyst is uniformly fixed in reaction channel.
Further, the ratio between amount of substance of the 6- nitrobenzimidazole and butyl acrylate is preferably 1:3~5, optimal
It is selected as 1:4.
Further, the reaction temperature is preferably 40~50 DEG C, and most preferably 45 DEG C.
Further, the reaction time is preferably 30~40min, most preferably 35min.
Reaction product of the invention can collect online, and gained reaction solution can be can be obtained by conventional post-processing approach
3- (5- nitrobenzimidazole) butyl propionate, 3- (6- nitrobenzimidazole) butyl propionate.The routine post-processing approach can be with
Be: the vacuum distillation of gained reaction solution removes solvent, and with 200-300 mesh silica gel wet method dress post, elution reagent is petroleum ether: acetic acid second
Ester volume ratio=4:1, wet process upper prop after obtained sample is dissolved with a small amount of elution reagent collect eluent, while TLC tracking is washed
The obtained eluent merging containing single product is evaporated, respectively obtains 6- nitrobenzimidazole class shown in formula 2 by de- process
5- nitrobenzimidazole analog derivative shown in derivative and formula 3.
Compared with prior art, the invention has the benefit that
The present invention utilizes 6- nitro benzo miaow shown in lipase-catalyzed online synthesis formula 2 in microfluidic channel reactor
5- nitrobenzimidazole analog derivative, the method shown in azole derivative and formula 3 not only significantly shorten the reaction time, but also
With high conversion ratio and selectivity；Glyoxaline compound is catalyzed using economic Lipozyme RM IM for the first time simultaneously
Michael reaction, reduce reaction cost, with economical and efficient advantage.
(4) Detailed description of the invention
Fig. 1 is the structural schematic diagram of microfluidic channel reactor used in the embodiment of the present invention.
In figure, 1,2- syringe, 3- reaction channel, 4- product collector, 5- constant temperature water box.
(5) specific embodiment
Protection scope of the present invention is described further with specific embodiment below, but protection scope of the present invention is unlimited
In this:
The structural reference Fig. 1 for the microfluidic channel reactor that the embodiment of the present invention uses, including a syringe pump (are not shown
Show), two syringes 1 and 2, reaction channel 3, constant temperature water box (5, only show its floor map) and product collector 4；Two
A syringe 1 and 2 is installed in syringe pump, is connect by a Y type interface with 3 entrance of reaction channel, the reaction channel 3 is set
In constant temperature water box 5, reaction temperature, the internal diameter 1.8mm of the reaction channel 3, pipe range are controlled by constant temperature water box 5
1m, the outlet of reaction channel 3 are connect by an interface with product collector 4.
Embodiment 1:3- (5- nitrobenzimidazole) butyl propionate, 3- (6- nitrobenzimidazole) butyl propionate
Device refers to Fig. 1: 6- nitrobenzimidazole (1.0mmol) being dissolved in 10mL DMSO, butyl acrylate
(4.0mmol) is dissolved in 10mL DMSO, then respectively loaded on spare in 10mL syringe.0.87g Lipozyme
RM IM is uniformly filled in reaction channel, and under the promotion of PHD2000 syringe pump, two-way reaction solution is respectively with 7.3 μ Lmin-1's
Flow velocity is entered in reaction channel by " Y " connector to be reacted, and is controlled temperature of reactor at 45 DEG C by constant temperature water box, is reacted
Liquid continuous flowing reactive 35min in reaction channel, reaction result pass through thin-layer chromatography TLC tracing detection.
Reaction solution is collected online by product collector, and vacuum distillation removes solvent, is filled with 200-300 mesh silica gel wet process
Column, elution reagent are petroleum ether: ethyl acetate=4:1, pillar height 35cm, column diameter 4.5cm, and sample is dissolved with a small amount of elution reagent
Wet process upper prop afterwards, eluent collect flow velocity 2mLmin-1, while TLC tracks elution process, will obtain containing single product
Eluent merging is evaporated, and obtains syrupy liq, obtains 3- (5- nitrobenzimidazole) butyl propionate, 3- (6- nitro benzo miaow
Azoles) butyl propionate, HPLC detection 6- nitrobenzimidazole conversion ratio 74%.
Nuclear-magnetism characterization result is as follows:
3-(6-nitro-benzimidazole-1-yl)-propionic acid Butyl ester and3-(5-
nitro-benzimidazole-1-yl)-propionic acid Butyl ester.Yellow oil；1H NMR
(500MHz,CDCl3): δ=8.89 (s, 1H, Ar-H), 8.75 (d, 1H, J=2.0Hz, Ar-H), 8.63 (s, 1H, Ar-H),
8.51 (d, 1H, J=2.0Hz, Ar-H), 8.30 (m, 2H, Ar-H), 7.96 (d, 1H, J=8.9Hz, Ar-H), 7.60 (d, 1H, J
=8.9Hz, Ar-H), 4.68 (m, 4H, NCH2),4.07(m,4H,OCH2), 2.98 (m, 4H, O=CCH2),1.67(m,4H,
OCH2CH2 ),1.28(m,4H,CH2 CH3), 0.90 (t, J=7.2Hz, 6H, CH2CH3 )；13C NMR(125MHz,CDCl3): δ=
170.36,147.35,146.57,144.30,140.94,137.04,132.28,119.71,119.44,119.21,116.49,
110.17,107.28,65.43,41.39,34.25,30.40,18.97,13.58.ESI–MS(m/z):292(M+1).
Embodiment 2-4
Change the temperature of microfluidic channel reactor, with embodiment 1, reaction result is as shown in table 1 for other:
Table 1: influence of the temperature to reaction
Table 1 the result shows that, when flow velocity be 7.3 μ Lmin-1, when the reaction time is 35min, react the liter with temperature
Height, conversion ratio is also significantly raised, and when reaction temperature reaches 45 DEG C, the conversion ratio of reaction is best, will at this time if continuing to heat up
It can cause the reduction of enzymatic activity, decrease so as to cause the conversion ratio and selectivity of reaction, so micro-fluidic in the present invention
The optimum response of 3- (5- nitrobenzimidazole) butyl propionate, 3- (6- nitrobenzimidazole) butyl propionate in micro passage reaction
Temperature is 45 DEG C.
Embodiment 5-8
On the basis of the dosage of 6- nitrobenzimidazole, change in micro-fluidic micro passage reaction 6- nitrobenzimidazole with
The ratio between amount of substrate materials of butyl acrylate controls temperature 45 C, other are with embodiment 1, and the results are shown in Table 2:
Influence of the table 2:6- nitrobenzimidazole with butyl acrylate substrate molar ratio to reacting
| Embodiment | Butyl acrylate: 6- nitrobenzimidazole molar ratio | Conversion ratio [%] |
| 5 | 1:1 | 21 |
| 6 | 2:1 | 50 |
| 7 | 3:1 | 66 |
| 1 | 4:1 | 74 |
| 8 | 5:1 | 72 |
Table 2 the result shows that, when flow velocity be 7.3 μ Lmin-1, the reaction time is 35min, and reaction temperature is 45 DEG C,
Reactor is using DMSO as organic solvent, and with the increase of reactant butyl acrylate, the conversion ratio of reaction is also increased as, and works as bottom
When object ratio 6- nitrobenzimidazole and butyl acrylate are 1:4, the conversion ratio of reaction is optimal, so micro-fluidic micro- logical in the present invention
The ratio between amount of best substrate materials is 1:4 in road reactor.
Embodiment 9-11
Change the reaction time of microfluidic channel reactor, with embodiment 1, reaction result is as shown in table 3 for other:
Table 3: influence of the reaction time to reaction
| Embodiment | Reaction time [min] | Conversion ratio [%] |
| 9 | 25 | 51 |
| 10 | 30 | 65 |
| 1 | 35 | 74 |
| 11 | 40 | 72 |
Table 3 the result shows that, reaction carry out 30min can be obtained 65% 3- (5- nitrobenzimidazole) butyl propionate,
3- (6- nitrobenzimidazole) butyl propionate, 6- nitrobenzimidazole are substantially converted into 3- (5- nitrobenzimidazole) propionic acid fourth
Ester, 3- (6- nitrobenzimidazole) butyl propionate.With the increase in reaction time, the conversion ratio of reaction is gradually increased, and works as reaction
When carrying out 35min, 3- (5- nitrobenzimidazole) butyl propionate, 3- (6- nitrobenzimidazole) butyl propionate conversion ratio can be with
Reach 74%, at this time if continuing to extend the reaction time, will lead to the reduction of reaction conversion ratio and selectivity instead, thus, it is micro-
3- (5- nitrobenzimidazole) butyl propionate, 3- (6- nitrobenzimidazole) butyl propionate synthesize most in flow control channel reactor
The good time is 35min.
Comparative example 1-4
Change the catalyst in micro-fluidic micro passage reaction, is changed to porcine pancreatic lipase PPL (comparative example 1), fat respectively
Enzyme Novozym 435 (comparative example 2), bacillus alkaline protease (comparative example 3), Lipozyme TL IM (comparative example 4),
It is with embodiment 1, and the results are shown in Table 4.
Table 4: influence of the different enzymes to reaction conversion ratio
| Comparative example | Enzyme source | Conversion ratio (%) |
| 1 | PPL | 20 |
| 2 | Novozym 435 | 16 |
| 3 | Bacillus alkaline protease | 26 |
| 4 | Lipozyme TL IM | 55 |
| Embodiment 1 | Lipozyme RM IM | 74 |
Table 4 the result shows that, the Michael addition reaction for enzymatic glyoxaline compound in microfluidic channel reactor
For, different enzymes has fairly obvious influence to reaction.Lipozyme TL IM catalysis reaction, 3- (5- nitro benzo miaow
Azoles) butyl propionate, 3- (6- nitrobenzimidazole) butyl propionate conversion ratio be 55%.And utilize the catalysis of Novozym 435 should
Reaction, 3- (5- nitrobenzimidazole) butyl propionate, 3- (6- nitrobenzimidazole) butyl propionate conversion ratio be only 16%.From
The result of table 4 is seen, for the Michael of enzymatic glyoxaline compound in micro-fluidic reactor, most effective catalyst is
Lipozyme RM IM, 3- (5- nitrobenzimidazole) butyl propionate, 3- (6- nitrobenzimidazole) butyl propionate
Conversion ratio is 74%.
Embodiment 12:3- (5- nitrobenzimidazole) propanoic acid tert-butyl ester, 3- (6- nitrobenzimidazole) propanoic acid tert-butyl ester
6- nitrobenzimidazole (1.0mmol) is dissolved in 10mL DMSO, tert-butyl acrylate (4.0mmol) dissolution
In 10mL DMSO, then respectively loaded on spare in 10mL syringe.0.87g Lipozyme RM IM is uniformly filled
In reaction channel, under the promotion of PHD2000 syringe pump, two-way reaction solution is respectively with 7.3 μ Lmin-1Flow velocity connect by " Y "
Head, which enters in reaction channel, to be reacted, and controls temperature of reactor at 45 DEG C by constant temperature water box, reaction solution is in reaction channel
Interior continuous flowing reactive 35min, reaction result pass through thin-layer chromatography TLC tracing detection.
Reaction solution is collected online by product collector, and vacuum distillation removes solvent, is filled with 200-300 mesh silica gel wet process
Column, elution reagent are petroleum ether: ethyl acetate=4:1, pillar height 35cm, column diameter 4.5cm, and sample is dissolved with a small amount of elution reagent
Wet process upper prop afterwards, eluent collect flow velocity 2mLmin-1, while TLC tracks elution process, will obtain containing single product
Eluent merging is evaporated, and obtains syrupy liq, obtains 3- (5- nitrobenzimidazole) propanoic acid tert-butyl ester, 3- (6- nitro benzo
Imidazoles) propanoic acid tert-butyl ester, HPLC detection 6- nitrobenzimidazole conversion ratio 59%.
Nuclear-magnetism characterization result is as follows:
3-(6-nitro-benzimidazole-1-yl)-propionic acid tert-butyl ester and3-
(5-nitro-benzimidazole-1-yl)-propionic acid tert-butyl ester.Yellow oil；1H
NMR(500MHz,CDCl3): δ=8.72 (d, 1H, J=2.0Hz, Ar-H), 8.42 (d, 1H, J=2.0Hz, Ar-H), 8.29-
8.21 (m, 4H, Ar-H), 7.88 (d, 1H, J=8.9Hz, Ar-H), 7.51 (d, 1H, J=8.9Hz, Ar-H), 4.55 (m, 4H,
NCH2), 2.84 (m, 4H, O=CCH2),1.39(s,18H,OC(CH3)3)；13C NMR(125MHz,CDCl3): δ=169.48,
147.91,146.77,143.98,142.94,137.57,132.78,120.53,118.90,118.21,117.17,109.61,
106.65,82.28,82.25,41.09,41.03,35.46,35.39,27.96.ESI–MS(m/z):292(M+1).
Embodiment 13-15
Change the temperature of microfluidic channel reactor, with embodiment 12, reaction result is as shown in table 5 for other:
Table 5: influence of the temperature to reaction
Table 5 the result shows that, when flow velocity be 7.3 μ Lmin-1, when the reaction time is 35min, react the liter with temperature
Height, conversion ratio is also significantly raised, and when reaction temperature reaches 45 DEG C, the conversion ratio of reaction is best, will at this time if continuing to heat up
It can cause the reduction of enzymatic activity, decrease so as to cause the conversion ratio and selectivity of reaction, so micro-fluidic in the present invention
3- (5- nitrobenzimidazole) propanoic acid tert-butyl ester in micro passage reaction, 3- (6- nitrobenzimidazole) propanoic acid tert-butyl ester it is best
Reaction temperature is 45 DEG C.
Embodiment 16-19
On the basis of the dosage of 6- nitrobenzimidazole, change in micro-fluidic micro passage reaction 6- nitrobenzimidazole with
The ratio between amount of substrate materials of tert-butyl acrylate controls temperature 45 C, other are with embodiment 12, and the results are shown in Table 6:
Influence of the table 6:6- nitrobenzimidazole with tert-butyl acrylate substrate molar ratio to reacting
Table 6 the result shows that, when flow velocity be 7.3 μ Lmin-1, the reaction time is 35min, and reaction temperature is 45 DEG C,
Reactor is using DMSO as organic solvent, and with the increase of reactant tert-butyl acrylate, the conversion ratio of reaction is also increased as, when
When substrate ratio 6- nitrobenzimidazole and tert-butyl acrylate are 1:4, the conversion ratio of reaction is optimal, so micro-fluidic in the present invention
The ratio between amount of best substrate materials is 1:4 in micro passage reaction.
Embodiment 20-11
Change the reaction time of microfluidic channel reactor, with embodiment 12, reaction result is as shown in table 7 for other:
Table 7: influence of the reaction time to reaction
| Embodiment | Reaction time [min] | Conversion ratio [%] |
| 20 | 25 | 39 |
| 21 | 30 | 51 |
| 12 | 35 | 59 |
| 22 | 40 | 57 |
Table 7 the result shows that, reaction carry out 30min can be obtained 51% the tertiary fourth of 3- (5- nitrobenzimidazole) propionic acid
Ester, 3- (6- nitrobenzimidazole) propanoic acid tert-butyl ester, 6- nitrobenzimidazole are substantially converted into 3- (5- nitrobenzimidazole)
Propanoic acid tert-butyl ester, 3- (6- nitrobenzimidazole) propanoic acid tert-butyl ester.With the increase in reaction time, the conversion ratio of reaction gradually increases
Add, when reaction carries out 35min, 3- (5- nitrobenzimidazole) propanoic acid tert-butyl ester, the tertiary fourth of 3- (6- nitrobenzimidazole) propionic acid
The conversion ratio of ester can achieve 59%, at this time if continuing to extend the reaction time, will lead to reaction conversion ratio and selectivity instead
Reduction, thus, 3- (5- nitrobenzimidazole) propanoic acid tert-butyl ester, 3- (6- nitrobenzimidazole) in microfluidic channel reactor
The Best Times of propanoic acid tert-butyl ester synthesis are 35min.
Comparative example 1-4
Change the catalyst in micro-fluidic micro passage reaction, is changed to porcine pancreatic lipase PPL (comparative example 1), fat respectively
Enzyme Novozym 435 (comparative example 2), bacillus alkaline protease (comparative example 3), Lipozyme TL IM (comparative example 4),
It is with embodiment 12, and the results are shown in Table 8.
Table 8: influence of the different enzymes to reaction conversion ratio
| Comparative example | Enzyme source | Conversion ratio (%) |
| 1 | PPL | 18 |
| 2 | Novozym 435 | 12 |
| 3 | Bacillus alkaline protease | 21 |
| 4 | Lipozyme TL IM | 38 |
| Embodiment 1 | Lipozyme RM IM | 59 |
Table 8 the result shows that, the Michael addition reaction for enzymatic glyoxaline compound in microfluidic channel reactor
For, different enzymes has fairly obvious influence to reaction.Lipozyme TL IM catalysis reaction, 3- (5- nitro benzo miaow
Azoles) propanoic acid tert-butyl ester, 3- (6- nitrobenzimidazole) propanoic acid tert-butyl ester conversion ratio be 38%.And it is urged using Novozym 435
Change the reaction, 3- (5- nitrobenzimidazole) propanoic acid tert-butyl ester, 3- (6- nitrobenzimidazole) propanoic acid tert-butyl ester conversion ratio only
It is 12%.It is most effective for the Michael of enzymatic glyoxaline compound in micro-fluidic reactor in terms of the result of table 8
Catalyst is Lipozyme RM IM, 3- (5- nitrobenzimidazole) propanoic acid tert-butyl ester, 3- (6- nitrobenzimidazole)
The conversion ratio of propanoic acid tert-butyl ester is 59%.
Claims (10)
1. a kind of method of lipase-catalyzed online synthesizing nitryl benzimidazoles derivative, it is characterised in that: the method is adopted
With microfluidic channel reactor, the microfluidic channel reactor includes syringe pump, syringe, reaction channel and collection of products
Device, the syringe are installed in syringe pump, are connected to by first interface with the entrance of the reaction channel, the collection of products
Device passes through the outlet of second interface and the reaction channel, and the reaction channel internal diameter is 0.8~2.4mm, reaction channel
A length of 0.5~1.0m；The described method includes: with the ratio between amount of substance for 1:1~5 6- nitrobenzimidazole and formula 1 shown in
Acrylics compound is raw material,, will using dimethyl sulfoxide as reaction dissolvent using Lipozyme RM IM as catalyst
The Lipozyme RM IM is uniformly filled in reaction channel, and raw material and reaction dissolvent are placed in syringe,
Raw material and reaction dissolvent are continuously passed through in the reaction channel under the promotion of syringe pump and carry out Michael by the syringe
Addition reaction, control reaction temperature are 40~55 DEG C, and the reaction time is 25~40min, are collected online by product collector anti-
Liquid is answered, the reaction solution is post-treated to respectively obtain 5- shown in 6- nitrobenzimidazole analog derivative and formula 3 shown in formula 2
Nitrobenzimidazole analog derivative；The catalysis interior, described to greatest extent of institute's catalyst filling can be accommodated in the reaction channel
The additional amount of agent is calculated as 0.025~0.05g/mL with the volume of the reaction dissolvent；In reaction system, third shown in the formula 1
The concentration of olefin(e) acid lipoid substance is 0.1~0.5mmol/mL,
In formula 1,2 and formula 3, the R is CH2CH2CH2CH3Or C (CH3)3。
2. the method for lipase-catalyzed online synthesizing nitryl benzimidazoles derivative as described in claim 1, feature exist
In: there are two the syringes, is the first syringe and the second syringe respectively, and first connecting pipe is Y type or T
Type pipeline, first syringe are connected to two interfaces of the Y type or T-type pipeline with the second syringe and lead to
The Y type or the parallel connection of T-type pipeline is crossed to connect with the reaction channel again.
3. the method for lipase-catalyzed online synthesizing nitryl benzimidazoles derivative as claimed in claim 2, feature exist
In: the method includes the following steps: third shown in 6- nitrobenzimidazole and formula 1 with the ratio between amount of substance for 1:1~5
Olefin(e) acid lipoid substance is raw material, is reaction with dimethyl sulfoxide using 0.5~1.0g Lipozyme RMIM as catalyst
The Lipozyme RM IM is uniformly filled in reaction channel by solvent, first dissolves 6- nitro with dimethyl sulfoxide
Benzimidazole is loaded in the first syringe, and the acrylics compound shown in dimethyl sulfoxide dissolution type 1 is loaded on the second injection
In device；First syringe, the second syringe are loaded in same syringe pump again, then in the synchronization of the syringe pump
Enter progress Michael in reaction channel after summarizing raw material and reaction dissolvent by the Y type or T-type pipeline to add
At reaction, controlling reaction temperature is 40~55 DEG C, and the reaction time is 25~40min, collects reaction online by product collector
Liquid, the reaction solution is post-treated to respectively obtain 5- nitre shown in 6- nitrobenzimidazole analog derivative and formula 3 shown in formula 2
Base benzimidazoles derivative；In reaction system, the concentration of acrylics compound shown in the formula 1 is 0.1~
0.5mmol/mL。
4. the method for lipase-catalyzed online synthesizing nitryl benzimidazoles derivative as described in claim 1, feature exist
In: the microfluidic channel reactor includes insulating box, and the reaction channel is placed in insulating box.
5. the method for lipase-catalyzed online synthesizing nitryl benzimidazoles derivative as claimed in claim 2, feature exist
In: the microfluidic channel reactor includes insulating box, and the reaction channel is placed in insulating box.
6. the method for the lipase-catalyzed online synthesizing nitryl benzimidazoles derivative as described in one of Claims 1 to 5,
Be characterized in that: the ratio between amount of substance of acrylics compound shown in the 6- nitrobenzimidazole and formula 1 is 1:3~5.
7. the method for the lipase-catalyzed online synthesizing nitryl benzimidazoles derivative as described in one of Claims 1 to 5,
Be characterized in that: the reaction temperature is 45~50 DEG C, and the reaction time is 30~40min.
8. the method for the lipase-catalyzed online synthesizing nitryl benzimidazoles derivative as described in one of Claims 1 to 5,
Be characterized in that: the ratio between amount of substance of acrylics compound shown in the 6- nitrobenzimidazole and formula 1 is 1:4.
9. the method for the lipase-catalyzed online synthesizing nitryl benzimidazoles derivative as described in one of Claims 1 to 5,
Be characterized in that: the reaction temperature is 45 DEG C, and the reaction time is 35min.
10. the method for the lipase-catalyzed online synthesizing nitryl benzimidazoles derivative as described in one of Claims 1 to 5,
It is characterized by: the post-processing approach of the reaction solution are as follows: the vacuum distillation of gained reaction solution removes solvent, and gained crude product is through silicon
Plastic column chromatography separation, with 200-300 mesh silica gel wet method dress post, elution reagent is the mixing of ethyl acetate, petroleum ether volume ratio 1:4
Solvent, wet process upper prop after obtained crude product is dissolved with a small amount of elution reagent collect eluent, while TLC tracks elution process,
To obtain the eluent merging containing single product be evaporated 6- nitrobenzimidazole analog derivative shown in the formula of respectively obtaining 2 and
5- nitrobenzimidazole analog derivative shown in formula 3.
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