Patent GB-1094566-A (GB)
Title: Kasugamycin and processes for the preparation thereof
IPC: C07H15/207
Filed: 1964-11-30
Abstract: An antibiotic substance, kasugamycin, is prepared by cultivating a strain of Streptomyces kasugaenois (ATCC. 15714 and 15715) or kasugamycin-producing variants and mutants thereof in an aqueous carbohydrate solution containing a nitrogenous nutrient under submerged aerobic conditions until substantial anti-bacterial activity is imparted to said solution and then recovering the antibiotic from the solution.  An aqueous solution of kasugamycin may be separated from the fermentation broth by adsorption on active carbon followed by elution with aqueous methanol, aqueous ethanol, aqueous acetone or water saturated with butanol, or on a cation exchange resin preferably having sulphonic acid as the active group followed by elution with aqueous ammonia. Kasugamycin may also be adsorbed on anion exchange resins treated with boric acid from which it may be eluted with aqueous hydrochloric acid.  Strong basic impurities may be removed by adsorption on a cation exchange resin having carboxylic acid groups.  The fermented broth and aqueous solutions of kasugamycin may be concentrated and dried, e.g. by distillation in vacuum, spray drying or freeze drying and the powder so obtained can be further purified by active carbon adsorption or solvent extraction.  Kasugamycin which is effective in inhibiting Brucella, Pseudomonas, Salmonella, Shigella, some Klebsiella and Blastmyces, effective in protection of rice plant from progressive infection of Piricularis oryzae is a weakly basic substance which melts at 207-216 DEG  C. under decomposition, is soluble in water, slightly soluble in methanol, substantially insoluble in ethanol, acetone, ethylacetate, ether, chloroform and benzene, exhibits no absorption maximum of U.V. light from 220 to 400 mm , gives a positive reaction to ninhydrin in pyridine and negative Tollens, Sakaguchi, Molisch, Fehlings, Elson-Morgan and ferric chloride reactions, has the empirical formula C14H25O9N3, contains a free primary amino group exhibiting pK1a less than 2.0, 7.1, 10.6 shows a titration equivalent of about 453, yields (+)-inositol by its acid hydrolysis, yields C14H24O10N2.H2O by its hydrolysis with baryta which further hydrolyses to C12H24O7N2 and oxalic acid, yields d-inositol (C6H12O6) and methyl glycoside (C9H17O4N3.HCl. 1/2 H2O) on methanolysis and forms acid addition salts, giving a crystalline kasugamycin hydrochloride which melts at 202-204 DEG  C. under decomposition is soluble in water, substantially insoluble in methanol, ethanol, acetone, ethyl acetate, chloroform and benzene, has the empirical formula C14H25O9N3.HCl.H2O exhibits (a )o25 + 120 DEG  (1.6% H2O), exhibits absorption bands in the infra-red region of the spectrum when pelleted in KBr at the following wave numbers in cm-1 3520, 3350, 3200, 3070, 2950, 2050, 1695, 1670, 1625, 1522, 1462, 1379, 1323, 1286, 1224, 1180, 1135, 1120, 1090, 1080, 1060, 1042, 1025, 975, 945, 908, 890, 870, 846, 825, 783 and 709 exhibits bands in the N.M.R. spectrum in D2O at the following p.p.m. 1.22, 1.32, 2.25, 2.33, 2.42, 2.50, 3.55, 3.79, 4.05, 4.38, 4.50, 4.70, 5.32 and 5.35, properties of kasugamycin hydrobromide being that it has the empirical formula C14H25O9N3.HBr.H2O, properties of kasugamycin sulphate being that it is soluble in water and decomposes at 210-218 DEG  C. The structure of kasugamycin has been determined as <FORM:1094566/C2/1>