CN111455004B - Method for synthesizing coumarin-3-carboxylic acid-6' -O-D-mannose ester on line by lipase catalysis - Google Patents
Method for synthesizing coumarin-3-carboxylic acid-6' -O-D-mannose ester on line by lipase catalysis Download PDFInfo
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Abstract
A method for synthesizing coumarin-3-carboxylic acid-6' -O-D-mannose ester on line by lipase catalysis, which comprises the following steps: uniformly filling lipase RM IM in a reaction channel of a microfluidic channel reactor, dissolving coumarin-3-carboxylic acid methyl ester and D-mannose respectively by using a reaction solvent, respectively injecting the solution into a pipeline through a first injector and a second injector for integration, then entering the reaction channel for reaction, controlling the reaction temperature to be 30-60 ℃, continuously flowing mixed solution in the reaction channel for 10-60 min, collecting the reaction solution flowing out of the reaction channel on line through a product collector, and performing aftertreatment to obtain the coumarin-3-carboxylic acid-6' -O-D-mannose ester; the method has the advantages of short reaction time, high yield and good selectivity.
Description
Technical Field
The invention relates to a method for synthesizing coumarin-3-carboxylic acid-6' -O-D-mannose ester on line by lipase catalysis.
Background
Coumarin is a basic structural subunit of a variety of plant secondary and microbial metabolites, and has a range of attractive biological activities including antibacterial, anticoagulant, antiviral, antitubercular, antioxidant, and antitumor activities. Several natural synthetic drugs containing coumarin scaffolds, such as warfarin as an anticoagulant for vitamin K antagonists, have been used clinically and widely in thrombosis treatment, and the common antibiotics, armillaria mellea a and novobiocin, both contain coumarin subunits. In addition, due to the parent structure of coumarin, it is widely used in the fields of specific fluorescent probes, dyes, fluorescent imaging, and the like. Among them, the synthesis of coumarin derivatives containing saccharide branches has attracted considerable attention in organic and pharmaceutical research and development. The interesting point of the coumarin containing saccharides is that these compounds have good water solubility. These results can improve the physicochemical, biopharmaceutical and pharmacokinetic properties of the drug. The study by sorsen et al showed that glycosylation of warfarin caused a shift from anticoagulant to anticancer activity, and they demonstrated that glycosylation of warfarin could show 70-fold higher anticancer activity than the original compound. This study clearly demonstrates that additional sugars are critical to altering the mechanism of action and potency of coumarin parent drugs. Over the past several years, several studies have been reported on coumarin containing sugars. Su Pulan et al synthesized a series of glycosyl coumarin carbonic anhydrase IX and XII inhibitors, which have strong inhibition effect on the growth of primary breast tumors. In 2016, nilsson et al reported a selective galectin-3 inhibitor, a coumarin derivative, which showed similar effects to the known non-selective galectin-1/galectin-3 inhibitor in a mouse model of bleomycin-induced pulmonary fibrosis.
The construction of sugar-containing derivatives can be achieved by basic synthetic methods, the most common synthetic strategy being chemical. The specific active hydroxyl groups on the saccharides are selectively synthesized by a "protection" or "protection deprotection" step. Visible light has also recently been reported as a glycosylation catalyst, however, most schemes for light-induced glycosylation require a transition metal catalyst in combination with expensive additives or oxidants to perform the reaction. Thus, the introduction of sugar chemically is still limited by the disadvantages of poor regio-and stereoselectivity, lengthy protection and deprotection of functional groups.
Biocatalysts have attracted considerable attention from chemists and biochemists as an efficient green bioconversion tool in organic synthesis. Particularly, the catalytic compounding in biocatalysis, namely, the old enzyme is utilized to form a new bond and follow a new path, so that the method has extremely and rapidly expanded. Some enzymes, such as engineered C-glycosyltransferase (micgtb-gagm), have been used in the synthesis of coumarin C-glycosides, where both C-glucosides synthesized have strong SGLT2 inhibitory activity. The enzyme-catalyzed reaction is relatively mild, green, but requires a long reaction time (typically as long as 24 hours or more) to achieve the desired result, and some rely on expensive enzymes. In recent years, continuous flow microreactors have become an effective way to shorten reaction time and increase yield by enzyme coupling.
Modern synthetic chemistry faces challenges in providing society with high performance, environmental protection, low cost, safety, and atomic efficiency valuable products. In this regard, continuous flow microreactor technology (MRT) is becoming increasingly popular as a replacement for traditional batch chemical synthesis. In particular, with respect to the 12-element principle of green chemistry, MRT can play a major role in improving chemical processes. The high surface to volume ratio of the microreactor means results in better heat exchange and efficient mixing, thus increasing the reaction efficiency. Furthermore, MRT systems have at the beginning of science involved reaction scales that allow efficient on-demand production of compounds in compact, reconfigurable equipment. In this case, "outwardly expanded" or "upwardly numbered" refers to a continuous flow system array operating in parallel to meet the desired output. Flow chemistry, particularly catalyst/substrate conditions in continuous flow systems, can improve reactivity and selectivity. At the same time, when the continuous flow column is filled with heterogeneous catalyst, separation of catalyst and product is very easy. In order to explore new, eco-friendly and efficient schemes for sugar-containing coumarin and as part of our ongoing research on the development of novel sugar-containing drugs, we have found a method for synthesizing coumarin-3-carboxylic acid-6 '-O-D-mannose esters on line by lipase catalysis in a microchannel reactor, aiming at finding an efficient and environment-friendly on-line controllable selective synthesis method for coumarin-3-carboxylic acid-6' -O-D-mannose esters.
Disclosure of Invention
The invention aims to provide a novel process method for synthesizing coumarin-3-carboxylic acid-6' -O-D-mannose ester on line by lipase catalysis in a microfluidic channel reactor, which has the advantages of short reaction time, high yield and good selectivity.
The technical scheme of the invention is as follows:
a method for synthesizing coumarin-3-carboxylic acid-6' -O-D-mannose ester on line by lipase catalysis, which comprises the following steps:
uniformly filling lipase RM IM (catalyst) into a reaction channel of a microfluidic channel reactor, dissolving coumarin-3-carboxylic acid methyl ester and D-mannose respectively by using a reaction solvent, respectively injecting the solution into a pipeline through a first injector and a second injector for integration, then entering the reaction channel for reaction, controlling the reaction temperature to be 30-60 ℃ (preferably 35 ℃), controlling the continuous flowing reaction time of a mixed solution in the reaction channel to be 10-60 min (preferably 40 min), collecting the reaction solution flowing out of the reaction channel on line through a product collector, and performing aftertreatment to obtain a product coumarin-3-carboxylic acid-6' -O-D-mannose ester (I);
the reaction solvent is a mixed solvent of dimethyl sulfoxide and tertiary amyl alcohol, wherein the volume ratio of dimethyl sulfoxide to tertiary amyl alcohol is 1:8 to 20, preferably 1:18;
the ratio of the mass of coumarin-3-carboxylic acid methyl ester to D-mannose in the mixed solution entering the reaction channel is 1:0.2 to 3, preferably 1:0.25, the specific operation can be as follows: after the coumarin-3-carboxylic acid methyl ester and the D-mannose are respectively dissolved by using a reaction solvent, the mass concentration ratio of substances of the coumarin-3-carboxylic acid methyl ester solution and the D-mannose solution is 1:0.2 to 3, preferably 1:0.25; the coumarin-3-carboxylic acid methyl ester solution and the D-mannose solution have the same flow rate when being injected by a first injector and a second injector respectively;
the lipase RM IM is commercially available, for example from Novozymes corporation, which is a preparation of 1, 3-position specific, food grade lipase (EC 3.1.1.3) on granular silica gel prepared from microorganisms produced by submerged fermentation with a genetically modified Aspergillus oryzae (Aspergillus oryzae) microorganism, obtained from Rhizomucor miehei; the lipase RM IM can be obtained by directly and uniformly fixing a granular catalyst in a reaction channel through a physical method; the catalyst is added in an amount of 0.025-0.05 g/mL based on the volume of the reaction medium within the maximum limit of the reaction channel capable of accommodating the filled catalyst;
the post-treatment method comprises the following steps: the solvent was removed by distillation under reduced pressure from the reaction mixture, and the mixture was subjected to column chromatography on a silica gel column, and the column was packed with 200-300 mesh silica gel by wet method, with the volume ratio of dichloromethane and methanol=10: 1.5, collecting the eluent containing the target compound by TLC tracking the elution process, evaporating the solvent and drying to obtain the product coumarin-3-carboxylic acid-6' -O-D-mannose ester (I);
the synthesis method adopts a microfluidic channel reactor, and the microfluidic channel reactor comprises the following steps: a first syringe, a second syringe, a reaction channel, and a product collector; the first injector and the second injector are connected with the inlet of the reaction channel through Y-shaped or T-shaped pipelines, and the product collector is connected with the outlet of the reaction channel through a pipeline;
further, the method comprises the steps of,
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 first injector and the second injector are arranged in the injection pump, and are synchronously pushed by the injection pump, and the specification of the first injector is consistent with that of the second injector;
the microfluidic channel reactor also comprises an incubator, wherein the reaction channel is arranged in the incubator, so that the reaction temperature can be effectively controlled, and the incubator can be selected according to the reaction temperature requirement, such as a water bath incubator and the like;
the material of the reaction channel is not limited, and green and environment-friendly materials such as silicone tubes are recommended; the shape of the reaction channel is preferably curved, so that the reaction liquid can pass through the reaction channel stably at a constant speed.
Compared with the prior art, the invention has the beneficial effects that:
the method utilizes lipase to catalyze and synthesize coumarin-3-carboxylic acid-6' -O-D-mannose ester on line in the microfluidic channel reactor, so that the method not only greatly shortens the reaction time, but also has high conversion rate and selectivity; meanwhile, the economic lipase RM IM is utilized for the first time to catalyze the reaction of coumarin-3-carboxylic acid methyl ester and D-mannose, so that the reaction cost is reduced, and the method has the advantages of economy and high efficiency.
Drawings
Fig. 1 is a schematic structural diagram of a microfluidic channel reactor according to an embodiment of the present invention.
In the figure, a 1-first syringe, a 2-second syringe, a 3-reaction channel, a 4-product collector and a 5-water bath incubator.
Detailed Description
The invention is further illustrated by the following examples, but the scope of the invention is not limited thereto:
the structure of the microfluidic channel reactor used in the embodiment of the present invention is shown in fig. 1, and includes an injection pump, a first injector 1, a second injector 2, a reaction channel 3, a water bath incubator 5 (only a schematic plan view thereof is shown), and a product collector 4; the first injector 1 and the second injector 2 are arranged in an injection pump and are connected with the inlet of a reaction channel 3 through a Y-shaped interface, the reaction channel 3 is arranged in a water bath constant temperature box 5, the reaction temperature is controlled through the water bath constant temperature box 5, the inner diameter of the reaction channel 3 is 2.0mm, the tube length is 1.0m, and the outlet of the reaction channel 3 is connected with a product collector 4 through an interface.
Example 1: synthesis of coumarin-3-carboxylic acid-6' -O-D-mannose ester
The apparatus is described with reference to fig. 1: coumarin-3-carboxylic acid methyl ester (2.0 mmol) was dissolved in 0.52mL dimethyl sulfoxide and 9.48mL t-amyl alcohol, D-mannose (0.5 mmol) was dissolved in 0.52mL dimethyl sulfoxide and 9.48mL t-amyl alcohol, and then each was filled into 10mL syringes for use. Uniformly filling 0.87g lipase RM IM in the reaction channel, and respectively mixing the two reaction solutions at a concentration of 7.8 μL.min under the drive of PHD 2000 injection pump -1 Is connected through 'Y' jointThe reaction head enters a reaction channel to react, the temperature of the reactor is controlled to be 35 ℃ by a water bath thermostat, the reaction liquid continuously reacts for 40 minutes in the reaction channel, and the reaction result is tracked and detected by thin layer chromatography TLC.
Collecting reaction liquid on line through a product collector, distilling under reduced pressure to remove solvent, loading into a column by using 200-300 mesh silica gel wet method, eluting with dichloromethane: methanol=10:1.5, column height of 35cm, column diameter of 4.5cm, dissolving sample with a small amount of eluting reagent, loading into the column by wet method, and collecting eluent with flow rate of 2 mL.min -1 And simultaneously, TLC tracks the elution process, the obtained eluates containing single products are combined and evaporated to dryness to obtain white solid, and coumarin-3-carboxylic acid-6 '-O-D-mannose ester is obtained, and the conversion rate of coumarin-3-carboxylic acid-6' -O-D-mannose ester is detected by HPLC and is 69%, and the selectivity is 99%.
The nuclear magnetic characterization results were as follows:
1 H NMR(DMSO-d 6 ,500MHz,δ,ppm)8.74(s,1H,H-4),7.89(dd,J＝7.8,1.6Hz,1H,H-5),7.76(ddd,J＝8.7,7.3,1.6Hz,1H,H-7),7.49–7.40(m,2H,H-8,H-6),6.41(d,J＝4.4Hz,1H,C1'-OH of D-mannose),5.01(d,J＝5.6Hz,1H,C1'-H of D-mannose),4.90(d,J＝4.4Hz,1H,C4'-OH of D-mannose),4.72(d,J＝4.1Hz,1H,C3'-OH of D-mannose),4.64(d,J＝5.5Hz,1H,C2'-OH of D-mannose),4.55(dd,J＝11.6,2.2Hz,1H,C6'-Ha of D-mannose),4.30(dd,J＝11.6,6.2Hz,1H,C6'-Hb of D-mannose),3.84(ddd,J＝8.7,6.2,2.1Hz,1H,C5'-H of D-mannose),3.63–3.56(m,2H,C2'-H,C3'-H of D-mannose),3.55(dd,J＝9.4,5.5Hz,1H,C4'-H of D-mannose). 13 C NMR(126MHz,DMSO)δ162.43(C-11),156.06(C-2),154.52(C-9),148.53(C-4),134.59(C-7),130.23(C-5),124.96(C-6),117.88(C-10),117.77(C-8),116.28(C-3),94.16(C-1'),71.36(C-2'),70.53(C-3'),70.34(C-5'),67.13(C-4'),65.16(C-6').
examples 2 to 8
The volume ratio of the reaction medium DMSO to the tertiary amyl alcohol in the microfluidic channel reactor was changed, the substrate ratio of coumarin-3-carboxylic acid methyl ester to D-mannose was 2:1 (1.0 mmol:0.5 mmol), the temperature was controlled to 50 ℃, the reaction time was 30min, the other same as in example 1, and the reaction results are shown in Table 1:
TABLE 1 influence of the volume ratio of DMSO to t-amyl alcohol in the reaction Medium on the reaction
| Examples | DMSO: tert-amyl alcohol | Conversion [%] | Selectivity [%] |
| 2 | 1:8 | n.d. | / |
| 3 | 1:10 | 14％ | 99％ |
| 4 | 1:12 | 23％ | 99％ |
| 5 | 1:14 | 30％ | 99％ |
| 6 | 1:16 | 38％ | 99％ |
| 7 | 1:18 | 40％ | 99％ |
| 8 | 1:20 | 36％ | 99％ |
The results in Table 1 show that when the molar ratio of the reactants coumarin-3-carboxylic acid methyl ester to D-mannose substrate is 2:1, the flow rate is 10.4. Mu.L.min -1 The reaction time is 30min, when the reaction temperature is 50 ℃, the conversion rate of the reaction is increased along with the increase of the volume ratio of the tertiary amyl alcohol in the reaction medium, and when the volume ratio of the DMSO in the reaction medium to the tertiary amyl alcohol is 1:18, the conversion rate of the reaction is optimal, and at the moment, if the volume ratio of the tertiary amyl alcohol is continuously increased, the dissolution amount of sugar in the reaction medium is reduced, so that the conversion rate of the reaction is reduced. The optimal reaction medium volume ratio for this reaction in the microfluidic microchannel reactor of the invention is thus DMSO: t-amyl alcohol=1:18.
Examples 9 to 15
The molar ratio of coumarin-3-carboxylic acid methyl ester to D-mannose substrate in the microfluidic microchannel reactor was changed, the reactor temperature was controlled at 50deg.C, the reaction time was 30min, and the results are shown in Table 2, except for example 1:
TABLE 2 influence of the ratio of the amounts of coumarin-3-carboxylic acid methyl ester and D-mannose substrate material on the reaction
| Examples | Coumarin-3-carboxylic acid methyl ester: d-mannose | Conversion [%] | Selectivity [%] |
| 9 | 5:1 | 50％ | 98％ |
| 10 | 4:1 | 51％ | 99％ |
| 11 | 3:1 | 47％ | 99％ |
| 12 | 2:1 | 40％ | 99％ |
| 13 | 1:1 | 32％ | 98％ |
| 14 | 1:2 | 30％ | 98％ |
| 15 | 1:3 | 24％ | 98％ |
The results in Table 2 show that the volume ratio of the reaction medium DMSO to t-amyl alcohol is 1:18, the flow rate is 10.4. Mu.L.min -1 The reaction time is 30min, the reaction conversion rate is increased along with the increase of the reactant coumarin-3-carboxylic acid methyl ester when the reaction temperature is 50 ℃, and the reaction conversion rate is optimal when the substrate ratio is 4:1, and the reaction conversion rate is reduced if the consumption of the reactant coumarin-3-carboxylic acid methyl ester is continuously increased. The optimal substrate molar ratio for this reaction in the microfluidic microchannel reactor of the invention is thus coumarin-3-carboxylic acid methyl ester to D-mannose=4:1.
Examples 16 to 22
The temperature of the microfluidic channel reactor was varied and the reaction time was controlled to 30min, otherwise the same as in example 1, and the reaction results are shown in table 3:
table 3: influence of temperature on the reaction
The results in Table 3 show that when the volume ratio of DMSO to t-amyl alcohol in the reaction medium is 1:18, the substrate molar ratio of coumarin-3-carboxylic acid methyl ester to D-mannose is 4:1, and the flow rate is 10.4. Mu.L.min -1 When the reaction time is 30min, the conversion rate of the reaction is optimal when the reaction temperature is 35 ℃, and the activity of the enzyme is affected by the temperature which is too high or too low. The optimum reaction temperature for this reaction in the microfluidic microchannel reactor of the invention is 35℃。
Examples 23 to 27
The reaction time of the microfluidic channel reactor was varied, and the reaction results are shown in table 4, except for example 1:
table 4: effect of reaction time on reaction
| Examples | Time [ min] | Conversion [%] | Selectivity [%] |
| 23 | 10 | 40％ | 99％ |
| 24 | 20 | 53％ | 99％ |
| 25 | 30 | 63％ | 99％ |
| 1 | 40 | 69％ | 99％ |
| 26 | 50 | 65％ | 99％ |
| 27 | 60 | 58％ | 95％ |
The results in Table 4 show that when the volume ratio of DMSO to t-amyl alcohol is 1:18, the substrate molar ratio of coumarin-3-carboxylic acid methyl ester to D-mannose is 4:1, and the reaction temperatures are 35℃at a flow rate of 7.8. Mu.L.min -1 When the reaction time is up to 40min, the reaction conversion rate can reach 69%, and if the reaction time is prolonged, the reaction conversion rate is reduced. Thus, the optimal reaction time for this reaction in a microfluidic channel reactor was 40min.
Comparative examples 1 to 3
The catalysts in the microfluidic microchannel reactor were changed to porcine pancreatic lipase PPL (comparative example 1), lipase Novozym 435 (comparative example 2), and subtilisin (comparative example 3), respectively, and the results are shown in table 5.
Table 5: influence of different enzymes on reaction conversion and selectivity
| Comparative example | Enzyme source | Conversion [%] | Selectivity [%] |
| 1 | PPL | 20％ | 74％ |
| 2 | Novozym 435 | 37％ | 73％ |
| 3 | Bacillus subtilis alkaline protease | 0 | 0 |
| Example 1 | Lipozyme RM IM | 69％ | 99％ |
The results in Table 5 show that for the regioselective transesterification of enzymatic coumarin-3-carboxylic acid methyl esters with D-mannose in a microfluidic reactor, different enzymes have a very pronounced effect on the reaction. The porcine pancreatic lipase PPL is utilized for catalytic reaction, and the conversion rate is 20%; catalyzing the reaction by using bacillus subtilis alkaline protease, wherein the conversion rate is 0%; the reaction was catalyzed with Novozym 435 with a conversion of 37%. From the results in table 5, the most efficient catalyst for the regioselective transesterification of enzymatic coumarin-3-carboxylic acid methyl ester with D-mannose in a microfluidic reactor was lipase RM IM with a conversion of 69% and a selectivity of 99%.
Comparative examples 4 to 6
The reaction materials in the microfluidic microchannel reactor were changed, the acyl donor was changed to tert-butyl coumarin-3-carboxylate (comparative example 4), the acyl acceptor was changed to D-galactose (comparative example 5), and the acyl donor and the acyl acceptor were changed (comparative example 6), and the results are shown in table 6, except that the reaction materials were changed to the same materials as in example 1.
TABLE 6 influence of different enzymes on reaction conversion and selectivity
| Comparative example | Acyl donor | Acyl acceptors | Conversion [%] | Selectivity [%] |
| 4 | Coumarin-3-carboxylic acid tert-butyl ester | D-mannose | 23％ | 84％ |
| 5 | Coumarin-3-carboxylic acid methyl ester | D-galactose | 8％ | 67％ |
| 6 | Coumarin-3-carboxylic acid tert-butyl ester | D-galactose | 0 | 0 |
| Example 1 | Coumarin-3-carboxylic acid methyl ester | D-mannose | 69％ | 99％ |
The results in Table 6 show that for the regioselective transesterification synthesis of enzymatically coumarin-3-carboxylic acid sugar esters in microfluidic reactors, the different substrates have a very pronounced effect on the reaction. Using coumarin-3-carboxylic acid tert-butyl ester as acyl donor and D-mannose as acyl acceptor, the conversion rate is 23%; using coumarin-3-carboxylic acid methyl ester as acyl donor, using D-galactose as acyl acceptor, and the conversion rate is 8%; the coumarin-3-carboxylic acid tert-butyl ester is used as an acyl donor, and the D-galactose is used as an acyl acceptor, so that the conversion rate is 0%. From the results of table 6, coumarin-3-carboxylic acid tert-butyl ester is not an effective acyl donor and D-galactose is not an effective acyl acceptor for the regioselective transesterification synthesis of enzymatic coumarin-3-carboxylic acid sugar esters in microfluidic reactors.
Application example 1
The size of the inhibition zone of coumarin-3-carboxylic acid-6' -O-D-mannose ester on staphylococcus aureus is measured by using an oxford cup method. 100. Mu.L of test bacteria solution (concentration of bacteria solution 1X 10) was added to the nutrient agar plate 7 CFU/mL), uniformly coating the bacterial liquid by using a sterile coater; 3 oxford cups are placed on the surface of the culture medium at equal distance and are pressed lightly, so that the oxford cups are contacted with the culture medium; 200. Mu.L (1 g/mL) of coumarin-3-carboxylic acid-6' -O-D-mannose ester was added to the cup and repeated 3 times; culturing in a 28 ℃ water-proof constant temperature incubator for 24 hours, and observing the result.
And (3) result judgment: the diameter of the bacteria growth zone is measured by a ruler by taking a bacteria growth zone which is not visible to naked eyes around the oxford cup as the bacteria inhibition zone, and the average value of 3 measurement results is taken as the bacteria inhibition zone of the compound to staphylococcus aureus. The diameter of the inhibition zone is expressed as d, and when d is less than 10mm, the inhibition zone is drug resistance (R); when d is more than or equal to 10 and less than or equal to 15, the sensor is moderately sensitive (I); when d >15mm, is highly sensitive (S).
TABLE 7 in vitro antibacterial test of coumarin-3-carboxylic acid-6' -O-D-mannite against Staphylococcus aureus
| Application example | Compounds of formula (I) | Average inhibition zone diameter/mm | Sensitivity to |
| 1 | Coumarin-3-carboxylic acid-6' -O-D-mannite | 16 | S |
Table 7 shows that coumarin-3-carboxylic acid-6' -O-D-mannitol has good inhibitory effect on Staphylococcus aureus, and can be used as Staphylococcus aureus inhibitor.
Claims (3)
1. A method for synthesizing coumarin-3-carboxylic acid-6' -O-D-mannose ester on line by lipase catalysis is characterized by comprising the following steps:
uniformly filling lipase RM IM in a reaction channel of a microfluidic channel reactor, dissolving coumarin-3-carboxylic acid methyl ester and D-mannose respectively by using a reaction solvent, respectively injecting the solution into a pipeline through a first injector and a second injector for integration, then entering the reaction channel for reaction, controlling the reaction temperature to be 30-60 ℃, continuously flowing mixed solution in the reaction channel for 10-60 min, collecting the reaction solution flowing out of the reaction channel on line through a product collector, and performing aftertreatment to obtain a product coumarin-3-carboxylic acid-6' -O-D-mannose ester (I);
the reaction solvent is a mixed solvent of dimethyl sulfoxide and tertiary amyl alcohol, wherein the volume ratio of dimethyl sulfoxide to tertiary amyl alcohol is 1: 8-20;
the ratio of the mass of coumarin-3-carboxylic acid methyl ester to D-mannose in the mixed solution entering the reaction channel is 1:0.2 to 3;
the post-treatment method comprises the following steps: collecting reaction liquid on line through a product collector, distilling under reduced pressure to remove solvent, loading into a column by using 200-300 mesh silica gel wet method, eluting with dichloromethane: methanol=10:1.5, column height of 35cm, column diameter of 4.5cm, loading into the column by using wet method after dissolving sample with eluting agent, and collecting eluent with flow rate of 2 mL.min -1 Simultaneously, TLC tracks the elution process, and the obtained eluents containing single products are combined and evaporated to dryness to obtain white solid, so as to obtain coumarin-3-carboxylic acid-6' -O-D-mannose ester;
2. the method for synthesizing coumarin-3-carboxylic acid-6' -O-D-mannose ester on line by lipase catalysis according to claim 1, wherein after the coumarin-3-carboxylic acid methyl ester and the D-mannose are respectively dissolved by a reaction solvent, the ratio of the mass concentration of the substance of the coumarin-3-carboxylic acid methyl ester solution to the mass concentration of the substance of the D-mannose solution is 1:0.2 to 3; the coumarin-3-carboxylic acid methyl ester solution and the D-mannose solution have the same flow velocity when being injected by the first injector and the second injector respectively.
3. The method for synthesizing coumarin-3-carboxylic acid-6' -O-D-mannose ester on line by lipase catalysis according to claim 1, wherein the catalyst lipase RM IM is added in an amount of 0.025-0.05 g/mL based on the volume of the reaction medium.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104593445A (en) * | 2015-01-21 | 2015-05-06 | 深圳大学 | Method for synthesizing sucrose fatty acid ester |
| CN109988794A (en) * | 2018-12-24 | 2019-07-09 | 浙江农林大学 | A method of the Enzyme catalyzed synthesis nitrobenzimidazole analog derivative in continuous flow reactor |
| CN109988787A (en) * | 2018-12-24 | 2019-07-09 | 浙江农林大学 | A kind of method of lipase-catalyzed online synthesis 2- phenylamino cyclohexanol |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104593445A (en) * | 2015-01-21 | 2015-05-06 | 深圳大学 | Method for synthesizing sucrose fatty acid ester |
| CN109988794A (en) * | 2018-12-24 | 2019-07-09 | 浙江农林大学 | A method of the Enzyme catalyzed synthesis nitrobenzimidazole analog derivative in continuous flow reactor |
| CN109988787A (en) * | 2018-12-24 | 2019-07-09 | 浙江农林大学 | A kind of method of lipase-catalyzed online synthesis 2- phenylamino cyclohexanol |
Non-Patent Citations (2)
| Title |
|---|
| Li-Hua Du等.A sustainable innovation for the tandem synthesis of sugar-containing coumarin derivatives catalyzed by lipozyme TL IM from Thermomyces lanuginosus in continuous-flow microreactors.《RSC Adv》.2020,第10卷(第22期),第13252-13259页. * |
| Li-Hua Du等.Continuous Flow Biocatalysis: Synthesis of Coumarin Carboxamide Derivatives by Lipase TL IM from Thermomyces lanuginosus.《Catalysts》.2022,第12卷(第3期),e339第1-14页. * |
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