ES252350A1 - The new organic compound tylosin, its hydrolysis product desmycosin and methods for preparing same - Google Patents

Abstract

A new antibiotic designated Tylosin is produced by cultivating a Tylosin-producing strain of Streptomyces fradiae in an aqueous culture medium containing assimilable sources of carbon, nitrogen and inorganic salts preferably under submerged aerobic conditions. The Tylosin produced gives Desmycosin and mycarose on acid hydrolysis with e.g. dilute hydrochloric acid. Specified strains of Streptomyces fradiae are S. Fradiae NRRL 2702 and S. Fradiae NRRL 2703. Specified sources of carbon are glucose, sucrose, fructose, starch, <PICT:0901273/IV (b)/1> <PICT:0901273/IV (b)/2> glycerin, molasses, dextrin, brown agar or corn steep solids. Specified sources of nitrogen are linseed meal, tankage, fish meal, cotton seed meal, oatmeal, ground wheat, soyabean meal, beef extract, peptones (meat or soy), casein or amino acid mixtures. Specified inorganic salts are those containing, sodium, potassium, ammonium, calcium, magnesium, cobalt, sulphate, chloride, phosphate, carbonate, acetate or nitrate ions. The aqueous nutrient medium may also contain growth factors e.g. distillers' solubles or yeast extract. Cultivation between 25 to 32 DEG C. for 2 to 5 days at an initial pH 5.5 to 8 is preferred. The antibiotic Tylosin is isolated from the clarified broth by extraction with a water-immiscible polar organis solvent such as chloroform or amyl acetate, or by adsorption on carbon, silica gel, alumina or acidic ion exchange resins such as "Dowex 50" (Registered Trade Mark) from chloroform benzene or acetone solution and elution with a low molecular weight alcohol e.g. methanol or a low molecular weight alcohol containing up to 50% of a low molecular weight ketone e.g. acetone. The Tylosin is further purified by concentrating the organic extract, decolorizing with carbon and precipitating with a non polar solvent e.g. petroleum ether or by adsorption chromatography of the organic extract. Both Tylosin and Desmycosin form acid addition salts with organic and inorganic acids and are readily acylated, (the acyl group containing 2-6 carbon atoms) they are soluble in polar organic solvents and relatively insoluble in non-polar solvents of the aliphatic hydrocarbon class and are white solids. They have a U.V. maxima at 288 mm , decolorize potassium permanganate, give positive Molisch and anthrone tests and negative Ninhydrin, biuret, Sakaguchi, ferric chloride and maltol tests. They have the following distinguishing properties:- <TABS:0901273/IV (b)/1> and I.R. spectrums as shown in Figs. I and II. The antibiotics Tylosin, Desmycosin, their salts and acylated derivatives, are active against gram positive and gram negative bacteria, they can be used as sterilizing agents and in veterinary medicines for the treatment of Turkey sinusitis.