A new antibiotic echanomycin, echanomycinic acid and process for their manufacture

摘要

PICT:0858210/IV(b)/1 A new antibiotic designated echanomycin is prepared by the aerobic p fermentation of a echanomycin-producing strain of streptomyces echinates nov. Spec. in a nutrient medium containing a source of nitrogen, carbohydrates and inorganic salts and preferably including growth promoting substances. Specified are amino-acids and mixtures thereof, peptides and proteins including their hydrolysates such as peptone or typtone, meat extracts, water-soluble constituents of cereal grains such as maize or wheat, distillers residues, soybean meal, cottonseed meal, glucose, lactose, saccharose and starch. Inorganic salts are chlorides, nitrates, carbonates and sulphates of alkali metals, alkaline earth metals, magnesium, iron, zinc or manganese. Fermentation is for 36 to 120 hours at 18 to 40 DEG C. Echanomycin is isolated from the clarified broth by (a) extraction or (b) adsorption. A small amount of echanomycin adsorbed on the mycelium is recovered by extraction with a partially water-soluble solvent such as acetone and this may be added to the clarified broth. Alternatively the whole broth may be extracted. Extraction is by organic solvents immiscible with water, e.g. esters of fatty acids such as ethyl or amyl acetate, hydrocarbons such as benzene, chlorinated hydrocarbons such as chloroform, ketones such as methyl propyl ketone, alcohols such as butyl alcohol, ethers and glycol ethers. The extract is then treated with (a) aqueous acid of pH below 5 e.g. acetic, hydrochloric or sulphuric, or citrate or phosphate buffers (b) aqueous alkaline solution of pH above 8, e.g. sodium or potassium hydroxide or phosphate buffers, the echanomycin being recovered from the organic layer. A further small recovery can be made from (b) by extraction with organic solvents below pH 5,0. Initially, or as a further step in purification, recovery may be by adsorption or e.g. active carbon or fuller's earth, followed by elution with a partially watermiscible solvent such as acetone. Further purification may be by means of (a) counter-current extraction between aqueous alcoholic solution and a water-immiscible organic solvent, e.g. carbon tetrachloride chloroform and aqueous methanol, or by chromatography on e.g. aluminium oxide. Echanomycin is colourless substance, melting at 217-218 DEG C., having specific rotation [a ]D = -310 DEG , analysis C = 55,79%, H = 5,74%, S = 5,24%, N (CH3) 5,20%, C (CH3) 2,63% sq possibly C29 H37 O7 N7 S; ultraviolet absorption spectrum bands at 242 mm FORM:0858210/IV(b)/1 ; improved spectrum at 5,74, 6,00, 6,81, 7,06, 7,22, 7,85-8,00, 8,75, 9,06, 12,78 and 13,16 microns. On acid hydrolysis, the amino acids d-serine and l-alanine are obtained together with a product giving a colour with rinhydrin. On alkaline hydrolysis, an acid, echanomycin acid is obtained. This produces quinoxaline carboxylic acid-(2) and ammonia on acid hydrolysis. Echanomycin is used medicinally (see Group VI)ALSO:A new antibiotic, echanomycia, produced by culturing streptomyces echinatics novspec (see Group IV (b)), active against a wide range of microorganisms including Trichomonas foctus, or its hydrolysic product echanomycinic acid, are formulated into pharmaceutical preparations in admixture with an organic or inorganic carrier suitable for enteral, parenteral or local administration. Suitable forms are tablets, dragees, salves, creams, powders, suppositories, solutions, suspensions or emulsions. Specified carriers are gelatine, lactose, starch, magnesium stearate, talc, vegetable oils, benzyl alcohols, gums, polyalkylene glycols, petroleum jelly and cholesterol. Preserving, stabilizing, wetting or emulsifying agents may be present together with other drugs.