CN109593804B - Method for quickly synthesizing nitrobenzimidazole derivative through enzyme catalysis - Google Patents
Method for quickly synthesizing nitrobenzimidazole derivative through enzyme catalysis Download PDFInfo
- Publication number
- CN109593804B CN109593804B CN201811584631.7A CN201811584631A CN109593804B CN 109593804 B CN109593804 B CN 109593804B CN 201811584631 A CN201811584631 A CN 201811584631A CN 109593804 B CN109593804 B CN 109593804B
- Authority
- CN
- China
- Prior art keywords
- reaction
- nitrobenzimidazole
- channel
- injector
- lipase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 43
- KRTDQDCPEZRVGC-UHFFFAOYSA-N 2-nitro-1h-benzimidazole Chemical class C1=CC=C2NC([N+](=O)[O-])=NC2=C1 KRTDQDCPEZRVGC-UHFFFAOYSA-N 0.000 title claims abstract description 21
- 238000006555 catalytic reaction Methods 0.000 title claims abstract description 14
- 230000002194 synthesizing effect Effects 0.000 title claims abstract description 12
- 102000004190 Enzymes Human genes 0.000 title abstract description 14
- 108090000790 Enzymes Proteins 0.000 title abstract description 14
- 238000006243 chemical reaction Methods 0.000 claims abstract description 155
- XPAZGLFMMUODDK-UHFFFAOYSA-N 6-nitro-1h-benzimidazole Chemical compound [O-][N+](=O)C1=CC=C2N=CNC2=C1 XPAZGLFMMUODDK-UHFFFAOYSA-N 0.000 claims abstract description 42
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract description 37
- 239000002994 raw material Substances 0.000 claims abstract description 30
- 230000035484 reaction time Effects 0.000 claims abstract description 28
- 108010048733 Lipozyme Proteins 0.000 claims abstract description 23
- 239000012295 chemical reaction liquid Substances 0.000 claims abstract description 21
- 239000003054 catalyst Substances 0.000 claims abstract description 19
- 239000007810 chemical reaction solvent Substances 0.000 claims abstract description 14
- 238000006845 Michael addition reaction Methods 0.000 claims abstract description 13
- 239000002904 solvent Substances 0.000 claims abstract description 7
- 239000000047 product Substances 0.000 claims description 18
- -1 acrylate compound Chemical class 0.000 claims description 15
- 238000003786 synthesis reaction Methods 0.000 claims description 15
- 238000011049 filling Methods 0.000 claims description 13
- 238000002347 injection Methods 0.000 claims description 13
- 239000007924 injection Substances 0.000 claims description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 12
- 230000015572 biosynthetic process Effects 0.000 claims description 11
- 239000003480 eluent Substances 0.000 claims description 8
- 238000010828 elution Methods 0.000 claims description 8
- 238000004809 thin layer chromatography Methods 0.000 claims description 8
- 108090001060 Lipase Proteins 0.000 claims description 7
- 239000003153 chemical reaction reagent Substances 0.000 claims description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- 239000004367 Lipase Substances 0.000 claims description 6
- 102000004882 Lipase Human genes 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 235000019421 lipase Nutrition 0.000 claims description 6
- 239000000741 silica gel Substances 0.000 claims description 6
- 229910002027 silica gel Inorganic materials 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 6
- 238000001704 evaporation Methods 0.000 claims description 4
- 239000003208 petroleum Substances 0.000 claims description 3
- 238000011068 loading method Methods 0.000 claims description 2
- 230000001360 synchronised effect Effects 0.000 claims description 2
- 239000012043 crude product Substances 0.000 claims 2
- 238000013375 chromatographic separation Methods 0.000 claims 1
- 239000012046 mixed solvent Substances 0.000 claims 1
- 238000012856 packing Methods 0.000 claims 1
- 150000001252 acrylic acid derivatives Chemical class 0.000 abstract 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 18
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 14
- 230000000052 comparative effect Effects 0.000 description 12
- 229940088598 enzyme Drugs 0.000 description 12
- 239000000758 substrate Substances 0.000 description 11
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 8
- FCCDDURTIIUXBY-UHFFFAOYSA-N lipoamide Chemical compound NC(=O)CCCCC1CCSS1 FCCDDURTIIUXBY-UHFFFAOYSA-N 0.000 description 8
- CQEYYJKEWSMYFG-UHFFFAOYSA-N butyl acrylate Chemical compound CCCCOC(=O)C=C CQEYYJKEWSMYFG-UHFFFAOYSA-N 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 150000002460 imidazoles Chemical class 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 108010084311 Novozyme 435 Proteins 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 230000002255 enzymatic effect Effects 0.000 description 6
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 5
- 229940040461 lipase Drugs 0.000 description 5
- ISXSCDLOGDJUNJ-UHFFFAOYSA-N tert-butyl prop-2-enoate Chemical compound CC(C)(C)OC(=O)C=C ISXSCDLOGDJUNJ-UHFFFAOYSA-N 0.000 description 5
- 108091005658 Basic proteases Proteins 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 239000000376 reactant Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- 240000006439 Aspergillus oryzae Species 0.000 description 2
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 2
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- 102000019280 Pancreatic lipases Human genes 0.000 description 2
- 108050006759 Pancreatic lipases Proteins 0.000 description 2
- 108090000787 Subtilisin Proteins 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 229940116369 pancreatic lipase Drugs 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000002035 prolonged effect Effects 0.000 description 2
- 108010031480 Artificial Receptors Proteins 0.000 description 1
- IVXGAZWSWCOZPW-UHFFFAOYSA-N CC(C)(C)OC(=O)CCN1C=NC2=C1C=C(C=C2)[N+](=O)[O-] Chemical compound CC(C)(C)OC(=O)CCN1C=NC2=C1C=C(C=C2)[N+](=O)[O-] IVXGAZWSWCOZPW-UHFFFAOYSA-N 0.000 description 1
- AWAFTONGILRBMM-UHFFFAOYSA-N CCCCOC(=O)CCN1C=NC2=C1C=C(C=C2)[N+](=O)[O-] Chemical compound CCCCOC(=O)CCN1C=NC2=C1C=C(C=C2)[N+](=O)[O-] AWAFTONGILRBMM-UHFFFAOYSA-N 0.000 description 1
- 229910004664 Cerium(III) chloride Inorganic materials 0.000 description 1
- 102100031375 Endothelial lipase Human genes 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- 238000006957 Michael reaction Methods 0.000 description 1
- 241000235403 Rhizomucor miehei Species 0.000 description 1
- 229910003101 Y(NO3)3·6H2O Inorganic materials 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000007259 addition reaction Methods 0.000 description 1
- 229940124599 anti-inflammatory drug Drugs 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 229940124623 antihistamine drug Drugs 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000012237 artificial material Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 230000003592 biomimetic effect Effects 0.000 description 1
- VYLVYHXQOHJDJL-UHFFFAOYSA-K cerium trichloride Chemical compound Cl[Ce](Cl)Cl VYLVYHXQOHJDJL-UHFFFAOYSA-K 0.000 description 1
- 238000003889 chemical engineering Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000002888 effect on disease Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 231100001261 hazardous Toxicity 0.000 description 1
- 239000000383 hazardous chemical Substances 0.000 description 1
- 208000021822 hypotensive Diseases 0.000 description 1
- 230000001077 hypotensive effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 229940079865 intestinal antiinfectives imidazole derivative Drugs 0.000 description 1
- 239000002608 ionic liquid Substances 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000013021 overheating Methods 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/10—Nitrogen as only ring hetero atom
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/18—Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/16—Microfluidic devices; Capillary tubes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
- C12M41/22—Heat exchange systems, e.g. heat jackets or outer envelopes in contact with the bioreactor walls
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Sustainable Development (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Clinical Laboratory Science (AREA)
- Physics & Mathematics (AREA)
- Thermal Sciences (AREA)
- Analytical Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses a method for quickly synthesizing nitrobenzimidazole derivatives through enzyme catalysis, which comprises the following steps: 6-nitrobenzimidazole and acrylate compounds with the mass ratio of 1: 1-5 are used as raw materials, lipase Lipozyme RM IM is used as a catalyst, DMSO solvent is used as a reaction solvent, and the lipase Lipozyme RM IM is uniformly filled in a reaction channel of a microfluidic channel reactor, wherein the inner diameter of the reaction channel of the microfluidic channel reactor is 0.8-2.4 mm, and the length of the reaction channel is 0.5-1.0 m; continuously introducing the raw materials and a reaction solvent into a reaction channel to carry out a Michael addition reaction, controlling the temperature of the Michael addition reaction to be 40-55 ℃ and the time of the Michael addition reaction to be 25-40 min, collecting reaction liquid on line, and carrying out conventional aftertreatment on the reaction liquid to obtain the nitrobenzimidazole derivative. The invention has the advantages of short reaction time, high selectivity and high yield.
Description
(I) technical field
The invention relates to a method for quickly synthesizing nitrobenzimidazole derivatives through enzyme catalysis.
(II) background of the invention
Imidazole is an important five-membered aromatic heterocycle, shows excellent biological activity due to the unique electrical-rich structure, and is widely applied to the fields of pesticides, medicines, artificial materials, artificial receptors, supramolecular ligands, biomimetic catalysts and the like. In particular, in medicinal chemistry, a large number of mature imidazole compounds are used clinically as anticancer drugs, antimicrobial drugs, anti-inflammatory drugs, antihistamine drugs, anti-neurogenic drugs, hypotensive drugs and the like, and have remarkable curative effects on diseases.
Michael addition, which is one of the important tools for forming C-C, C-N, C-O, C-X bonds, is the classical addition reaction in organic synthesis. The Michael addition reaction is generally carried out under catalysis of strong acid or strong base, and the reaction conditions are harsh, so that the environmental pollution and the energy waste can be caused. In order to find a more efficient synthesis method, a series of metal catalysts such as KF/Al have been developed in recent years2O3、Y(NO3)3·6H2O、CeCl3、Bi(OTf)3Etc., but these catalysts produce environmentally harmful substances with accompanying many side reactions, seriously lowering the selectivity of the reaction and the yield of the objective product. In addition, the reaction is reported to be carried out using a solid supported catalyst, an ionic liquid, or the like, but these reactions have disadvantages such as a long reaction time and a complicated preparation process of the catalyst system. Therefore, the search for a new green synthesis technology of Michael addition reaction is a research hotspot in the field of organic synthesis.
The enzyme catalysis reaction is one of effective tools for green chemical synthesis due to high efficiency, green and strong specificity. Its excellent selectivity and mild reaction conditions have led to a wide interest of scientists in the fields of chemistry and chemical engineering, pharmacy, materials and the like. However, the enzymatic reaction has the restriction of solvent to substrate dissolution, solvent polarity to enzyme activity inhibition and the like, the reaction time is usually very long (24-96h), and the conversion rate of a specific substrate is not very high, so that the development of a novel synthesis technology of the enzymatic imidazole compound based on the micro-fluidic technology on the basis of the traditional enzymatic reaction becomes the research target of the people.
In recent years, continuous flow reactors with channel sizes in the micrometer or millimeter range have been widely used in organic synthesis. One of the most significant advantages of microfluidic reactors over conventional chemical reactors is the safety of the use of hazardous reagents. The method is derived from the high specific surface area and the better heat and mass transfer performance of the microreactor, and the local overheating phenomenon is obviously inhibited. At the same time, the smaller dimensions of the microreactor prevent the build-up of hazardous substances inside the reactor. Therefore, microreactor technology provides a unique means of performing ultra-fast exothermic reactions and allows reactions to be performed via highly unstable and even explosive intermediates.
So far, the enzymatic Michael addition reaction in China is in the initial research stage, and has certain defects, and the biological method mostly uses acylase for catalysis, is expensive, needs longer reaction time (24-96h), and is not particularly ideal for the conversion rate of specific substrate reaction. In order to develop a new efficient and green technology for synthesizing imidazole derivatives, a method for synthesizing nitrobenzimidazole derivatives on line under catalysis of lipase in a microchannel reactor is researched, and the new efficient and environment-friendly technology for synthesizing the nitrobenzimidazole derivatives on line is sought.
Disclosure of the invention
The technical problem to be solved by the invention is to provide a novel process for synthesizing the nitrobenzimidazole derivative on line by lipase catalysis in a microfluidic channel reactor, and the novel process has the advantages of short reaction time and high yield.
In order to solve the technical problems, the invention adopts the following technical scheme:
a method for synthesizing nitrobenzimidazole derivatives on line under catalysis of lipase adopts a microfluidic channel reactor, wherein the microfluidic channel reactor comprises an injection pump, an injector, a reaction channel and a product collector, the injector is installed in the injection pump and is communicated with an inlet of the reaction channel through a first interface, the product collector is communicated with an outlet of the reaction channel through a second interface, the inner diameter of the reaction channel is 0.8-2.4 mm, and the length of the reaction channel is 0.5-1.0 m; the method comprises the following steps: the method comprises the steps of uniformly filling a reaction channel with 6-nitrobenzimidazole (RBB) and an acrylate compound shown in a formula 1 as raw materials, lipase Lipozyme RM IM as a catalyst and dimethyl sulfoxide as a reaction solvent, placing the raw materials and the reaction solvent into an injector, continuously introducing the raw materials and the reaction solvent into the reaction channel by the injector under the driving of an injection pump to carry out Michael addition reaction, controlling the reaction temperature to be 40-55 ℃ and the reaction time to be 25-40 min, collecting the reaction liquid on line through a product collector, and respectively obtaining 6-nitrobenzimidazole derivatives shown in a formula 2 and 5-nitrobenzimidazole derivatives shown in a formula 3 through aftertreatment of the reaction liquid; the catalyst is added in an amount of 0.025-0.05 g/mL based on the volume of the reaction solvent within the maximum limit that the reaction channel can accommodate the filled catalyst; in the reaction system, the concentration of the acrylate compound shown in the formula 1 is 0.1-0.5 mmol/mL,
in the formulas 1, 2 and 3, R is CH2CH2CH2CH3Or C (CH)3)3。
Further, the present invention adopts a microfluidic channel reactor, wherein the number of the injectors can be one or more, depending on the specific reaction requirements. The reaction raw materials of the invention are two, preferably two injectors are used, specifically, the injectors are respectively a first injector and a second injector, the first connecting pipeline is a Y-shaped or T-shaped pipeline, the first injector and the second injector are respectively connected with two interfaces of the Y-shaped or T-shaped pipeline and are connected with the reaction channel in series through the Y-shaped or T-shaped pipeline, and the probability of contact and collision of reactant molecules passing through the microchannel is increased, so that two reactant liquid flows are mixed and react in the common reaction channel.
Still further, more specifically, the method of the present invention comprises the steps of:
the method comprises the following steps: taking 6-nitrobenzimidazole and an acrylate compound shown in a formula 1 in a mass ratio of 1: 1-5 as raw materials, taking 0.5-1.0 g of lipase Lipozyme RM IM as a catalyst, taking dimethyl sulfoxide as a reaction solvent, uniformly filling the lipase Lipozyme RM IM in a reaction channel, firstly dissolving the 6-nitrobenzimidazole in the dimethyl sulfoxide, filling the 6-nitrobenzimidazole in a first injector, and dissolving the acrylate compound shown in the formula 1 in the dimethyl sulfoxide, and filling the acrylate compound in a second injector; then, the first injector and the second injector are arranged in the same injection pump, then under the synchronous pushing of the injection pump, raw materials and a reaction solvent are converged through the Y-shaped or T-shaped pipeline and then enter a reaction channel for Michael addition reaction, the reaction temperature is controlled to be 40-55 ℃, the reaction time is 25-40 min, reaction liquid is collected on line through a product collector, and the reaction liquid is subjected to post-treatment to respectively obtain the 6-nitrobenzimidazole derivative shown in the formula 2 and the 5-nitrobenzimidazole derivative shown in the formula 3; in the reaction system, the concentration of the acrylate compound shown in the formula 1 is 0.1-0.5 mmol/mL.
The first syringe and the second syringe have the same specification, and the concentration of the 6-nitrobenzimidazole in the first syringe is usually 0.1 mmol/mL.
Furthermore, the microfluidic channel reactor also comprises a thermostat, and the reaction channel is arranged in the thermostat, so that the reaction temperature can be effectively controlled. The constant temperature box can be selected according to the reaction temperature requirement, such as a water bath constant temperature box and the like.
The material of the reaction channel is not limited, and green and environment-friendly materials such as a silicone tube are recommended; the shape of the reaction channel is preferably curved, so that the reaction liquid can be ensured to stably pass through at a constant speed.
In the present invention, the lipase Lipozyme RM IM is a preparation prepared from microorganisms, using a commercial product manufactured by Novozymes (novozymes), which is a food-grade lipase (EC 3.1.1.3) specific to 1, 3-position on granular silica gel. It is produced by submerged fermentation using a genetically modified Aspergillus oryzae (Aspergillus oryzae) microorganism obtained from Rhizomucor miehei.
The method of the invention uniformly fills the lipase Lipozyme RM IM in the reaction channel, and can directly and uniformly fix the granular catalyst in the reaction channel by a physical method.
Further, the mass ratio of the 6-nitrobenzimidazole to butyl acrylate is preferably 1:3 to 5, and most preferably 1: 4.
Further, the reaction temperature is preferably 40-50 ℃, and most preferably 45 ℃.
Further, the reaction time is preferably 30 to 40min, and most preferably 35 min.
The reaction product can be collected on line, and the obtained reaction liquid can be used for obtaining 3- (5-nitrobenzimidazole) butyl propionate and 3- (6-nitrobenzimidazole) butyl propionate by a conventional post-treatment method. The conventional post-treatment method may be: and (2) distilling the obtained reaction liquid under reduced pressure to remove the solvent, filling the reaction liquid into a column by using a 200-mesh 300-mesh silica gel wet method, wherein the volume ratio of an eluting reagent to ethyl acetate is 4:1, dissolving the obtained sample by using a small amount of an eluting reagent, then loading the sample into the column by using the wet method, collecting eluent, tracking the elution process by using TLC (thin layer chromatography), merging the obtained eluents containing a single product, and evaporating to dryness to obtain the 6-nitrobenzimidazole derivative shown in the formula 2 and the 5-nitrobenzimidazole derivative shown in the formula 3 respectively.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, the 6-nitrobenzimidazole derivative shown in the formula 2 and the 5-nitrobenzimidazole derivative shown in the formula 3 are synthesized on line in the microfluidic channel reactor by using lipase catalysis, so that the method not only greatly shortens the reaction time, but also has high conversion rate and selectivity; meanwhile, the economical lipase Lipozyme RM IM is used for catalyzing the Michael reaction of the imidazole compounds for the first time, so that the reaction cost is reduced, and the method has the advantages of economy and high efficiency.
(IV) description of the drawings
Fig. 1 is a schematic structural diagram of a microfluidic channel reactor used in an embodiment of the present invention.
In the figure, 1, 2-injector, 3-reaction channel, 4-product collector, 5-water bath incubator.
(V) detailed description of the preferred embodiments
The scope of the invention is further illustrated by the following examples, but is not limited thereto:
referring to fig. 1, a microfluidic channel reactor used in an embodiment of the present invention includes a syringe pump (not shown), two syringes 1 and 2, a reaction channel 3, a water bath incubator (5, only a schematic plan view thereof is shown), and a product collector 4; two injectors 1 and 2 are installed in the injection pump and are connected with an inlet of a reaction channel 3 through a Y-shaped interface, the reaction channel 3 is arranged in a water bath thermostat 5, the reaction temperature is controlled through the water bath thermostat 5, the inner diameter of the reaction channel 3 is 1.8mm, the length of a tube is 1m, and an outlet of the reaction channel 3 is connected with a product collector 4 through an interface.
Example 1: 3- (5-Nitrobenzimidazole) propionic acid butyl ester, 3- (6-nitrobenzimidazole) propionic acid butyl ester
The device is shown in figure 1: 6-nitrobenzimidazole (1.0mmol) was dissolved in 10mL DMSO and butyl acrylate (4.0mmol) was dissolved in 10mL DMSO, each in a 10mL syringe. 0.87g of lipase Lipozyme RM IM is uniformly filled in the reaction channel, and two paths of reaction liquid are respectively 7.3 mu L/min under the driving of a PHD2000 injection pump-1The flow rate of the reaction solution enters a reaction channel through a Y joint for reaction, the temperature of the reactor is controlled at 45 ℃ through a water bath thermostat, the reaction solution continuously and continuously reacts in the reaction channel for 35min, and the reaction result is tracked and detected through thin-layer chromatography TLC.
Collecting reaction liquid on line through a product collector, distilling under reduced pressure to remove a solvent, filling the reaction liquid into a column by using a 200-mesh 300-mesh silica gel wet method, dissolving a sample in a small amount of an elution reagent, namely petroleum ether and ethyl acetate with the ratio of 4:1, wherein the column height is 35cm, the column diameter is 4.5cm, filling the sample into the column by using the wet method, and collecting eluent at the flow rate of 2 mL/min-1And simultaneously tracking the elution process by TLC (thin layer chromatography), merging the obtained eluents containing the single product and evaporating to obtain syrup-shaped liquid, so as to obtain 3- (5-nitrobenzimidazole) butyl propionate and 3- (6-nitrobenzimidazole) butyl propionate, wherein the conversion rate of 6-nitrobenzimidazole is 74% by HPLC (high performance liquid chromatography).
The nuclear magnetic characterization results were 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,OCH2C 2H),1.28(m,4H,C 2HCH3),0.90(t,J＝7.2Hz,6H,CH2C 3H)；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).
examples 2 to 4
The temperature of the microfluidic channel reactor was changed, and the reaction results are shown in table 1 as in example 1:
table 1: influence of temperature on the reaction
The results in Table 1 show that when the flow rate is 7.3. mu.L.min-1When the reaction time is 35min, the conversion rate is obviously increased along with the increase of the reaction temperature, when the reaction temperature reaches 45 ℃, the conversion rate of the reaction is optimal, and if the temperature is continuously increased, the enzyme activity is reduced, so that the conversion rate and the selectivity of the reaction are reduced, and therefore, the optimal reaction temperature of the butyl 3- (5-nitrobenzimidazole) propionate and the optimal reaction temperature of the butyl 3- (6-nitrobenzimidazole) propionate in the microfluidic microchannel reactor are 45 ℃.
Examples 5 to 8
The results are shown in Table 2, except that the temperature is controlled at 45 ℃ in the microfluidic microchannel reactor by changing the ratio of the 6-nitrobenzimidazole to the butyl acrylate substrate in reference to the 6-nitrobenzimidazole dosage, and the results are shown in the same manner as in example 1:
TABLE 2 influence of the molar ratio of 6-nitrobenzimidazole to butyl acrylate substrate on the reaction
| Examples | Butyl acrylate: 6-nitrobenzimidazole molar ratio | Conversion [% ]] |
| 5 | 1:1 | 21 |
| 6 | 2:1 | 50 |
| 7 | 3:1 | 66 |
| 1 | 4:1 | 74 |
| 8 | 5:1 | 72 |
The results in Table 2 show that the flow rate was 7.3. mu.L.min-1The reaction time is 35min, the reaction temperature is 45 ℃, DMSO is used as an organic solvent in the reactor, the conversion rate of the reaction is increased along with the increase of butyl acrylate serving as a reactant, and when the substrate ratio of 6-nitrobenzimidazole to butyl acrylate is 1:4, the conversion rate of the reaction is optimal, so that the ratio of the optimal substrate amount in the microfluidic microchannel reactor is 1: 4.
Examples 9 to 11
The reaction time of the microfluidic channel reactor was changed, and the reaction results are shown in Table 3 as in example 1:
table 3: influence of reaction time on the reaction
| Examples | Reaction time [ min ]] | Conversion [% ]] |
| 9 | 25 | 51 |
| 10 | 30 | 65 |
| 1 | 35 | 74 |
| 11 | 40 | 72 |
The results in Table 3 show that 65% of butyl 3- (5-nitrobenzimidazole) propionate and butyl 3- (6-nitrobenzimidazole) propionate can be obtained after 30min of reaction, and 6-nitrobenzimidazole is basically converted into butyl 3- (5-nitrobenzimidazole) propionate and butyl 3- (6-nitrobenzimidazole) propionate. The conversion rate of the reaction is gradually increased along with the increase of the reaction time, when the reaction is carried out for 35min, the conversion rate of the butyl 3- (5-nitrobenzimidazole) propionate and the butyl 3- (6-nitrobenzimidazole) propionate can reach 74%, and at the moment, if the reaction time is continuously prolonged, the reduction of the conversion rate and the selectivity of the reaction is caused, so that the optimal time for synthesizing the butyl 3- (5-nitrobenzimidazole) propionate and the butyl 3- (6-nitrobenzimidazole) propionate in the microfluidic channel reactor is 35 min.
Comparative examples 1 to 4
The results are shown in Table 4 for the same samples as example 1 except that the catalysts in the microfluidic microchannel reactor were changed to porcine pancreatic lipase PPL (comparative example 1), lipase Novozym 435 (comparative example 2), subtilisin alkaline protease (comparative example 3), Lipozyme TL IM (comparative example 4), respectively.
TABLE 4 Effect of different enzymes on the conversion of the reaction
| Comparative example | Enzyme source | Conversion (%) |
| 1 | PPL | 20 |
| 2 | Novozym 435 | 16 |
| 3 | Bacillus subtilis alkaline protease | 26 |
| 4 | Lipozyme TL IM | 55 |
| Example 1 | Lipozyme RM IM | 74 |
The results in table 4 show that for the Michael addition reaction of enzymatic imidazoles in microfluidic channel reactors, different enzymes have a very significant effect on the reaction. The Lipozyme TL IM catalyzed reaction, the conversion rate of 3- (5-nitrobenzimidazole) butyl propionate and 3- (6-nitrobenzimidazole) butyl propionate is 55%. The conversion of butyl 3- (5-nitrobenzimidazole) propionate and butyl 3- (6-nitrobenzimidazole) propionate was only 16% when Novozym 435 catalyzed the reaction. From the results in table 4, the most effective catalyst for Michael of enzymatic imidazoles in microfluidic reactors was the lipase Lipozyme RM IM, with a conversion of 74% for butyl 3- (5-nitrobenzimidazole) propionate, butyl 3- (6-nitrobenzimidazole) propionate.
Example 12: tert-butyl 3- (5-nitrobenzimidazole) propionate, tert-butyl 3- (6-nitrobenzimidazole) propionate
6-Nitrobenzimidazole (1.0mmol) was dissolved in 10mL of DMSO, tert-butyl acrylate (4.0mmol) was dissolved in 10mL of DMSO, and the solutions were separately filled in 10mL syringesAnd (4) preparing for later use. 0.87g of lipase Lipozyme RM IM is uniformly filled in the reaction channel, and two paths of reaction liquid are respectively 7.3 mu L/min under the driving of a PHD2000 injection pump-1The flow rate of the reaction solution enters a reaction channel through a Y joint for reaction, the temperature of the reactor is controlled at 45 ℃ through a water bath thermostat, the reaction solution continuously and continuously reacts in the reaction channel for 35min, and the reaction result is tracked and detected through thin-layer chromatography TLC.
Collecting reaction liquid on line through a product collector, distilling under reduced pressure to remove a solvent, filling the reaction liquid into a column by using a 200-mesh 300-mesh silica gel wet method, dissolving a sample in a small amount of an elution reagent, namely petroleum ether and ethyl acetate with the ratio of 4:1, wherein the column height is 35cm, the column diameter is 4.5cm, filling the sample into the column by using the wet method, and collecting eluent at the flow rate of 2 mL/min-1And simultaneously tracking the elution process by TLC (thin layer chromatography), merging the obtained eluents containing the single product and evaporating to obtain syrup-shaped liquid, so as to obtain the tert-butyl 3- (5-nitrobenzimidazole) propionate and the tert-butyl 3- (6-nitrobenzimidazole) propionate, and detecting the conversion rate of the 6-nitrobenzimidazole by HPLC (high performance liquid chromatography) to be 59%.
The nuclear magnetic characterization results were 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).
examples 13 to 15
The temperature of the microfluidic channel reactor was changed, and the reaction results are shown in Table 5 as in example 12:
table 5: influence of temperature on the reaction
The results in Table 5 show that the flow rate was 7.3. mu.L.min-1When the reaction time is 35min, the reaction is obviously improved along with the increase of the temperature, the conversion rate of the reaction is optimal when the reaction temperature reaches 45 ℃, and if the temperature is continuously increased, the enzyme activity is reduced, so that the conversion rate and the selectivity of the reaction are reduced, and the optimal reaction temperature of the tert-butyl 3- (5-nitrobenzimidazole) propionate and the tert-butyl 3- (6-nitrobenzimidazole) propionate in the microfluidic microchannel reactor is 45 ℃.
Examples 16 to 19
The results are shown in Table 6, except that the temperature is controlled at 45 ℃ in the microfluidic microchannel reactor by changing the ratio of the substrate substances of 6-nitrobenzimidazole and tert-butyl acrylate based on the amount of 6-nitrobenzimidazole, and the results are the same as in example 12:
TABLE 6 influence of the molar ratio of 6-nitrobenzimidazole to tert-butyl acrylate substrate on the reaction
The results in Table 6 show that the flow rate was 7.3. mu.L.min-1The reaction time is 35min, the reaction temperature is 45 ℃, DMSO is used as an organic solvent in the reactor, the conversion rate of the reaction is increased along with the increase of the reactant tert-butyl acrylate, and when the substrate ratio of 6-nitrobenzimidazole to tert-butyl acrylate is 1:4, the conversion rate of the reaction is optimal, so that the ratio of the optimal substrate amount in the microfluidic microchannel reactor is 1: 4.
Examples 20 to 11
The reaction time of the microfluidic channel reactor was changed, and the reaction results are shown in Table 7 as in example 12:
table 7: influence of reaction time on the reaction
| Examples | Reaction time [ min ]] | Conversion [% ]] |
| 20 | 25 | 39 |
| 21 | 30 | 51 |
| 12 | 35 | 59 |
| 22 | 40 | 57 |
The results in Table 7 show that reaction time of 30min gave 51% of tert-butyl 3- (5-nitrobenzimidazole) propionate and tert-butyl 3- (6-nitrobenzimidazole) propionate, with the 6-nitrobenzimidazole being essentially converted to tert-butyl 3- (5-nitrobenzimidazole) propionate and tert-butyl 3- (6-nitrobenzimidazole) propionate. The conversion rate of the reaction gradually increases along with the increase of the reaction time, when the reaction is carried out for 35min, the conversion rate of the tert-butyl 3- (5-nitrobenzimidazole) propionate and the tert-butyl 3- (6-nitrobenzimidazole) propionate can reach 59%, and at the moment, if the reaction time is continuously prolonged, the reduction of the conversion rate and the selectivity of the reaction can be caused, so that the optimal time for synthesizing the tert-butyl 3- (5-nitrobenzimidazole) propionate and the tert-butyl 3- (6-nitrobenzimidazole) propionate in the microfluidic channel reactor is 35 min.
Comparative examples 1 to 4
The results are shown in Table 8 for the same samples as example 12 except that the catalysts in the microfluidic microchannel reactor were changed to porcine pancreatic lipase PPL (comparative example 1), lipase Novozym 435 (comparative example 2), subtilisin alkaline protease (comparative example 3), Lipozyme TL IM (comparative example 4), respectively.
TABLE 8 Effect of different enzymes on the conversion of the reaction
| Comparative example | Enzyme source | Conversion (%) |
| 1 | PPL | 18 |
| 2 | Novozym 435 | 12 |
| 3 | Bacillus subtilis alkaline protease | 21 |
| 4 | Lipozyme TL IM | 38 |
| Example 1 | Lipozyme RM IM | 59 |
The results in table 8 show that for the Michael addition reaction of enzymatic imidazoles in microfluidic channel reactors, different enzymes have a very significant effect on the reaction. The Lipozyme TL IM catalyzed reaction, the conversion rate of 3- (5-nitrobenzimidazole) butyl propionate and 3- (6-nitrobenzimidazole) butyl propionate is 38%. The reaction was catalyzed by Novozym 435, and the conversion of tert-butyl 3- (5-nitrobenzimidazole) propionate and tert-butyl 3- (6-nitrobenzimidazole) propionate was only 12%. From the results in Table 8, the most effective catalyst for Michael of enzymatic imidazoles in microfluidic reactors is the lipase Lipozyme RM IM, with 59% conversion of tert-butyl 3- (5-nitrobenzimidazole) propionate, tert-butyl 3- (6-nitrobenzimidazole) propionate.
Claims (10)
1. A method for synthesizing nitrobenzimidazole derivatives on line by lipase catalysis is characterized in that: the method adopts a microfluidic channel reactor, wherein the microfluidic channel reactor comprises an injection pump, an injector, a reaction channel and a product collector, the injector is arranged in the injection pump and is communicated with an inlet of the reaction channel through a first interface, the product collector is communicated with an outlet of the reaction channel through a second interface, the inner diameter of the reaction channel is 0.8-2.4 mm, and the length of the reaction channel is 0.5-1.0 m; the method comprises the following steps: the method comprises the steps of uniformly filling a reaction channel with 6-nitrobenzimidazole (RBB) and an acrylate compound shown in a formula 1 as raw materials, lipase Lipozyme RM IM as a catalyst and dimethyl sulfoxide as a reaction solvent, placing the raw materials and the reaction solvent into an injector, continuously introducing the raw materials and the reaction solvent into the reaction channel by the injector under the driving of an injection pump to carry out Michael addition reaction, controlling the reaction temperature to be 40-55 ℃ and the reaction time to be 25-40 min, collecting the reaction liquid on line through a product collector, and respectively obtaining 6-nitrobenzimidazole derivatives shown in a formula 2 and 5-nitrobenzimidazole derivatives shown in a formula 3 through aftertreatment of the reaction liquid; the catalyst is added in an amount of 0.025-0.05 g/mL based on the volume of the reaction solvent within the maximum limit that the reaction channel can accommodate the filled catalyst; in the reaction system, the concentration of the acrylate compound shown in the formula 1 is 0.1-0.5 mmol/mL,
in the formulas 1, 2 and 3, R is CH2CH2CH2CH3Or C (CH)3)3。
2. The method for the lipase-catalyzed on-line synthesis of nitrobenzimidazole derivatives according to claim 1, which comprises the following steps: the device comprises a reaction channel, a Y-shaped or T-shaped pipeline, two injectors, a first injector and a second injector, wherein the two injectors are respectively a first injector and a second injector, the first interface is a Y-shaped or T-shaped pipeline, the first injector and the second injector are respectively connected with the two interfaces of the Y-shaped or T-shaped pipeline, are connected in parallel through the Y-shaped or T-shaped pipeline and are then connected in series with the reaction channel.
3. The method for the lipase-catalyzed on-line synthesis of nitrobenzimidazole derivatives according to claim 2, which comprises the following steps: the method comprises the following steps: taking 6-nitrobenzimidazole and an acrylate compound shown in a formula 1 in a mass ratio of 1: 1-5 as raw materials, taking 0.5-1.0 g of lipase Lipozyme RM IM as a catalyst, taking dimethyl sulfoxide as a reaction solvent, uniformly filling the lipase Lipozyme RM IM in a reaction channel, firstly dissolving the 6-nitrobenzimidazole in the dimethyl sulfoxide, filling the 6-nitrobenzimidazole in a first injector, and dissolving the acrylate compound shown in the formula 1 in the dimethyl sulfoxide, and filling the acrylate compound in a second injector; then, the first injector and the second injector are arranged in the same injection pump, then under the synchronous pushing of the injection pump, raw materials and a reaction solvent are converged through the Y-shaped or T-shaped pipeline and then enter a reaction channel for Michael addition reaction, the reaction temperature is controlled to be 40-55 ℃, the reaction time is 25-40 min, reaction liquid is collected on line through a product collector, and the reaction liquid is subjected to post-treatment to respectively obtain the 6-nitrobenzimidazole derivative shown in the formula 2 and the 5-nitrobenzimidazole derivative shown in the formula 3; in the reaction system, the concentration of the acrylate compound shown in the formula 1 is 0.1-0.5 mmol/mL.
4. The method for the lipase-catalyzed on-line synthesis of nitrobenzimidazole derivatives according to claim 1, which comprises the following steps: the microfluidic channel reactor comprises a thermostat, and the reaction channel is arranged in the thermostat.
5. The method for the lipase-catalyzed on-line synthesis of nitrobenzimidazole derivatives according to claim 2, which comprises the following steps: the microfluidic channel reactor comprises a thermostat, and the reaction channel is arranged in the thermostat.
6. The method for the lipase-catalyzed on-line synthesis of nitrobenzimidazole derivatives according to any of claims 1 to 5, which comprises the following steps: the mass ratio of the 6-nitrobenzimidazole to the acrylate compound shown in the formula 1 is 1: 3-5.
7. The method for the lipase-catalyzed on-line synthesis of nitrobenzimidazole derivatives according to any of claims 1 to 5, which comprises the following steps: the reaction temperature is 45-50 ℃, and the reaction time is 30-40 min.
8. The method for the lipase-catalyzed on-line synthesis of nitrobenzimidazole derivatives according to any of claims 1 to 5, which comprises the following steps: the mass ratio of the 6-nitrobenzimidazole to the acrylate compound shown in the formula 1 is 1: 4.
9. The method for the lipase-catalyzed on-line synthesis of nitrobenzimidazole derivatives according to any of claims 1 to 5, which comprises the following steps: the reaction temperature is 45 ℃ and the reaction time is 35 min.
10. The method for the lipase-catalyzed on-line synthesis of nitrobenzimidazole derivatives according to any of claims 1 to 5, which comprises the following steps: the post-treatment method of the reaction solution comprises the following steps: and distilling the obtained reaction liquid under reduced pressure to remove the solvent, carrying out chromatographic separation on the obtained crude product by using a silica gel column, carrying out wet column packing by using 200-mesh 300-mesh silica gel, wherein an elution reagent is a mixed solvent of ethyl acetate and petroleum ether in a volume ratio of 1:4, dissolving the obtained crude product by using a small amount of elution reagent, carrying out wet column loading, collecting eluent, tracking an elution process by TLC (thin layer chromatography), combining the obtained eluent containing a single product, and evaporating to dryness to obtain the 6-nitrobenzimidazole derivative shown in the formula 2 and the 5-nitrobenzimidazole derivative shown in the formula 3 respectively.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811584631.7A CN109593804B (en) | 2018-12-24 | 2018-12-24 | Method for quickly synthesizing nitrobenzimidazole derivative through enzyme catalysis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811584631.7A CN109593804B (en) | 2018-12-24 | 2018-12-24 | Method for quickly synthesizing nitrobenzimidazole derivative through enzyme catalysis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109593804A CN109593804A (en) | 2019-04-09 |
| CN109593804B true CN109593804B (en) | 2021-06-08 |
Family
ID=65964384
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811584631.7A Active CN109593804B (en) | 2018-12-24 | 2018-12-24 | Method for quickly synthesizing nitrobenzimidazole derivative through enzyme catalysis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109593804B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111560408B (en) * | 2020-02-29 | 2022-11-25 | 浙江工业大学 | Method for synthesizing coumarin-3-carboxylic acid sugar ester derivative on line based on flow chemistry enzymatic catalysis |
| CN111690699A (en) * | 2020-05-19 | 2020-09-22 | 浙江工业大学 | Method for enzymatic on-line synthesis of 3- (pyridine-2-amino) propylhydroxamic acid |
| CN111690698A (en) * | 2020-05-19 | 2020-09-22 | 浙江工业大学 | Method for synthesizing 3- (benzo [ d ] [1,3] dioxo-5-amino) propyl hydroxamic acid by two-step serial flow |
| CN111690695A (en) * | 2020-05-19 | 2020-09-22 | 浙江工业大学 | Enzymatic microfluidic online synthesis method of 3-phenylaminopropylhydroxamic acid |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6667384B2 (en) * | 2001-12-27 | 2003-12-23 | Hercules Incorporated | Methyl acrylate-diamine based polyamide resins and processes for producing the same |
| CN101892273B (en) * | 2010-06-13 | 2013-01-09 | 西南大学 | Application of immobilized lipase Lipozyme TLIM as catalyst for unsymmetrical Michael addition reaction |
| CN104561173B (en) * | 2015-01-14 | 2018-06-26 | 浙江工业大学 | A kind of method of lipase-catalyzed online synthesis palmitic acid 1- (6- nitrobenzimidazoles base) ethyl ester |
| CN104561170B (en) * | 2015-01-14 | 2018-02-13 | 浙江工业大学 | A kind of method of lipase-catalyzed online synthesis of acetic acid 1 (6 nitrobenzimidazole base) ethyl ester |
| CN104561169B (en) * | 2015-01-14 | 2018-02-09 | 浙江工业大学 | A kind of method of lipase-catalyzed online synthesis laurate 1 (6 nitrobenzimidazole base) ethyl ester |
| CN104561171B (en) * | 2015-01-14 | 2018-06-26 | 浙江工业大学 | A kind of method of lipase-catalyzed online synthesizing adipic acid vinyl list 1- (6- nitrobenzimidazoles base) ethyl ester |
-
2018
- 2018-12-24 CN CN201811584631.7A patent/CN109593804B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN109593804A (en) | 2019-04-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109988794B (en) | Method for synthesizing nitrobenzimidazole derivatives through enzyme catalysis in continuous flow reactor | |
| CN109593804B (en) | Method for quickly synthesizing nitrobenzimidazole derivative through enzyme catalysis | |
| CN109706198A (en) | A kind of method that online enzyme process closes nitro imidazole derivatives | |
| CN109735582B (en) | Method for synthesizing cyclohexanol beta-amino alcohol derivatives on line by lipase catalysis | |
| CN107384991B (en) | Method for synthesizing 5' -O-ethylene adipamide uridine on line by lipase catalysis | |
| CN107488683A (en) | A kind of lipase-catalyzed online synthesis N（5 vinyl acetate valeryls）The method of mexiletine | |
| CN107988277B (en) | Method for synthesizing S-benzylpalmitic acid thioester on line under catalysis of lipase | |
| CN107384782A (en) | A kind of method of lipase-catalyzed online synthesis 5 '-O- ethene adipyls -5-FUD | |
| CN109706194B (en) | Method for synthesizing phenethyl alcohol beta-amino alcohol derivatives on line based on mobile chemical enzymatic ammonolysis reaction | |
| CN109988787B (en) | Method for synthesizing 2-phenylamino cyclohexanol on line under catalysis of lipase | |
| CN104561170B (en) | A kind of method of lipase-catalyzed online synthesis of acetic acid 1 (6 nitrobenzimidazole base) ethyl ester | |
| CN111690698A (en) | Method for synthesizing 3- (benzo [ d ] [1,3] dioxo-5-amino) propyl hydroxamic acid by two-step serial flow | |
| CN109762853B (en) | Method for synthesizing isopropanol beta-alkamine derivative on line by lipase catalysis | |
| CN107988278B (en) | Method for synthesizing S-benzyl lauric acid thioester on line by lipase catalysis | |
| CN104561174B (en) | A kind of method of lipase-catalyzed online synthesis palmitic acid 1 (4 nitroimidazole base) ethyl ester | |
| CN111455004B (en) | Method for synthesizing coumarin-3-carboxylic acid-6' -O-D-mannose ester on line by lipase catalysis | |
| CN115478086A (en) | Method for enzymatic on-line synthesis of adenine Michael addition derivatives | |
| CN115992192A (en) | Method for synthesizing benzimidazole Michael addition derivative in continuous flow reactor | |
| CN111676255A (en) | Method for synthesizing 3-phenylaminopropylhydroxamic acid on line under catalysis of lipase | |
| CN111690695A (en) | Enzymatic microfluidic online synthesis method of 3-phenylaminopropylhydroxamic acid | |
| CN104561175B (en) | A kind of method of lipase-catalyzed online synthesis 1- (4- nitro-imidazols base)-ethyl acetate | |
| CN104611388B (en) | A kind of method of lipase-catalyzed online synthesizing adipic acid vinyl list 1- (4- nitroimidazoles base) ethyl ester | |
| CN104561171B (en) | A kind of method of lipase-catalyzed online synthesizing adipic acid vinyl list 1- (6- nitrobenzimidazoles base) ethyl ester | |
| CN104561169B (en) | A kind of method of lipase-catalyzed online synthesis laurate 1 (6 nitrobenzimidazole base) ethyl ester | |
| CN104561173B (en) | A kind of method of lipase-catalyzed online synthesis palmitic acid 1- (6- nitrobenzimidazoles base) ethyl ester |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |