Structure of 1220993-44-1
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The BI-3802 was designed by Boehringer Ingelheim and could be obtained free of charge through the Boehringer Ingelheim open innovation portal opnMe.com, associated with its negative control.
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CAS No. : | 1220993-44-1 |
Formula : | C11H12O4 |
M.W : | 208.21 |
SMILES Code : | O=C([C@@H]1C2=CC(OC)=C(OC)C=C2C1)O |
MDL No. : | MFCD28133921 |
* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
35%Chromat. | With nitrilase of Rhodococcus rhodochrous NCIMB11216; In aq. phosphate buffer; dimethyl sulfoxide; at 30℃; for 96h;pH 7.3;Enzymatic reaction; | A study using a number of bacterial inducers (propionitrile, benzonitrile, 4-bromobenzonitrile) showed that propionitrile provided the best induction of nitrilase activity with <strong>[35202-54-1]3,4-dimethoxybicyclo[4.2.0]octa-1,3,5-triene-7-carbonitrile</strong>. (0103) The bacterial strains were induced with propionitrile at 72 mM for 72 hours, and the cells were taken up in 50 mL (twice concentrated, conc. 10 mg of cells per ml) of 0.1M phosphate buffer KH2PO4/K2HPO4 pH=7.3 and <strong>[35202-54-1]3,4-dimethoxybicyclo[4.2.0]octa-1,3,5-triene-7-carbonitrile</strong> was added at a concentration of 10 mM in 2percent of DMSO v/vfinal. (0104) The fungal strains were induced with valeronitrile. (0105) All the reaction mixtures were stirred at 220 rpm at 30° C. in the case of the bacteria and at 27° C. in the case of the fungi and monitored for 96 hours by reverse-phase HPLC and by chiral-phase HPLC according to the methods described below: Reverse-Phase Analysis Chiral-Phase Analysis (0107) Chiralpak IC 250*4.6 column (0108) 30percent absolute ethanol+0.1percent TFA+70percent heptane+0.1percent TFA (0109) 1 ml/min 30° C. 288 nm The results obtained are collated in the Table below: |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
36%; 39% | With over-expressed nitrilase of Rhodococcus rhodochrous NCIMB 11216; In aq. phosphate buffer; dimethyl sulfoxide; at 30℃; for 6h;pH 7.0;Enzymatic reaction; | Over-Expression of the Nitrilase: (0060) The nitrilase protein of Rhodococcus rhodochrous NCIMB 11216 is described in protein and genome databases. The sequence of the sought gene is listed under the identifier SVA (Sequence Version Archive) ?EF467367? in the ENA (European Nucleotide Archive) of EMBL-Bank. This sequence corresponds to the reference ?A4LA85? in the UniProtKB/TrEMBL database. (0061) The production strain E. coli BL21(DE3), transformed with the expression vector pET28a-Nit1, was used. (0062) The nitrilase over-expression protocol is described in Applied Biochemistry and Biotechnology 2010, Vol 160(2), pp 393-400. (0063) The cells thereby transformed are either used directly in the form of a bacterial slurry or are lyophilised before use. Enzymatic Hydrolysis Using the Over-Expressed Nitrilase. (0064) The cells transformed according to the above protocol are stirred at a concentration of 5.6×109 cells/mL (1 mL of culture at OD=1 (600 nm) corresponds to 1·109 bacteria and about 10 mg of bacterial slurry or 1.5 mg of lyophilisate). (0065) To a solution of 250 mL of phosphate buffer KH2PO4/Na2HPO4 1/15 M at pH 7 there are added 1 g of lyophilisate of E. coli and 500 mg (c=2 g/L, 10 mM) of <strong>[35202-54-1]3,4-dimethoxybicyclo[4.2.0]octa-1,3,5-triene-7-carbonitrile</strong> in 2percent of DMSO (5 mL). (0066) The reaction mixture is maintained at 30° C., with rotary stirring at 220 rpm, for 6 hours. (0067) The reaction is monitored by chiral-phase HPLC under conditions allowing the enantiomeric excess of the acid and nitrile to be determined: Chiralpak IB Column 90percent n-hexane 10percent 2-PrOH+0.1percent TFA 1 mL/min 30° C. 288 nm The chiral-phase HPLC chromatogram after 6 hours is shown in FIG. 1. (0070) After reacting for 6 hours, the reaction mixture is acidified with 1M HCl in order to obtain a highly acid pH (pH 2) and is then extracted with 2×100 mL of dichloromethane. The organic phase is drawn off. A second extraction using toluene (2×100 mL) makes it possible to recover all the product remaining in the aqueous phase. The organic phases are washed with saturated NaCl solution and then dried using anhydrous magnesium sulphate. After evaporation of the solvents, the crude product is obtained, which is purified by flash chromatography on a silica column under the following conditions: Column type: 80 g SiOH Macherey-Nagel Material and method: Reveleris Eluant: Isocratic (cyclohexane+1percent acetic acid/ethyl acetate+1percent acetic acid 75/25) Detection: UV 288 nm Flow rate: 60 ml/min Result: Nitrile (R): yield 36percent (179 mg), ee (R): 96percent Acid (S): yield 39percent (246 mg), ee (S): 96percent |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
35%Chromat.; 42%Chromat. | With nitrilase of Rhodococcus erythropolis NCIMB11215; In aq. phosphate buffer; dimethyl sulfoxide; at 30℃; for 96h;pH 7.3;Enzymatic reaction; | A study using a number of bacterial inducers (propionitrile, benzonitrile, 4-bromobenzonitrile) showed that propionitrile provided the best induction of nitrilase activity with <strong>[35202-54-1]3,4-dimethoxybicyclo[4.2.0]octa-1,3,5-triene-7-carbonitrile</strong>. (0103) The bacterial strains were induced with propionitrile at 72 mM for 72 hours, and the cells were taken up in 50 mL (twice concentrated, conc. 10 mg of cells per ml) of 0.1M phosphate buffer KH2PO4/K2HPO4 pH=7.3 and <strong>[35202-54-1]3,4-dimethoxybicyclo[4.2.0]octa-1,3,5-triene-7-carbonitrile</strong> was added at a concentration of 10 mM in 2percent of DMSO v/vfinal. (0104) The fungal strains were induced with valeronitrile. (0105) All the reaction mixtures were stirred at 220 rpm at 30° C. in the case of the bacteria and at 27° C. in the case of the fungi and monitored for 96 hours by reverse-phase HPLC and by chiral-phase HPLC according to the methods described below: Reverse-Phase Analysis Chiral-Phase Analysis (0107) Chiralpak IC 250*4.6 column (0108) 30percent absolute ethanol+0.1percent TFA+70percent heptane+0.1percent TFA (0109) 1 ml/min 30° C. 288 nm The results obtained are collated in the Table below: |
Tags: 1220993-44-1 synthesis path| 1220993-44-1 SDS| 1220993-44-1 COA| 1220993-44-1 purity| 1220993-44-1 application| 1220993-44-1 NMR| 1220993-44-1 COA| 1220993-44-1 structure
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