METHOD FOR PREPARATION OF 7-(N-BENZYLOXYCARBONYL)ARGINYLAMIDO-4-METHYLCOUMARINE

摘要

Invention applies to bio-organic chemistry, mainly the production of substrates used to evaluate activity of ferments and in the research of their activity mechanisms. The method for obtaining of N-benzyloxycarbonyl arginine 4-methylcoumaril-7 amide has been developed. This is substrate of fluorogenic ferments, the action of which is based on fermentative truncation of 7-amino-4-methylcoumarine and its ability to fluoresce at 440 nm.The method offered by the authors allows it to obtain N-benzyloxycarbonyl arginine 4 methylcoumaril-7 amide with high outcome (83%). The process is technologically easy. The result is achieved by mixing 7-amino-4-methylcoumarine suspension in pyridine together with phosphorous trichloride in pyridine for one hour after which hydrogen chloride of N-benzyloxycarbonyl arginine dissolved in pyridine is added. The mixture is heated for 3-4 hours at 40-50 degrees C temperature and kept for 10 hours in ambient temperature, pyridine is distilled off, the substance is treated with normal hydrochloric acid and crystallised from methanol/dimethylformamide (4:1).