Patent GB-762736-A (GB)
Title: The new antibiotic b-mycin and processes for its preparation
Filed: 1954-11-11
Abstract: A new antibiotic, B-mycin, is produced by cultivating Streptomyces bottropensis nov. spec. or a mutant thereof aerobically in an aqueous nutrient solution containing both carbon and nitrogen sources, under stationary or submerged conditions, and recovering the so-produced B-mycin from the fermentation broth.  The temperature is 20 DEG  to 35 DEG  C. and preferably 26 DEG  to 28 DEG  C., the pH 5 to 8 and preferably <PICT:0762736/IV(b)/1> <PICT:0762736/IV(b)/2> 6.5 to 8, and the duration of cultivation 3 to 7 days.  Specified sources of carbon are glucose, saccharose, lactose, starch and sugar alcohols, especially glycerol.  Specified sources of nitrogen are hydrolysed or non-hydrolysed casein, corn steep liquor, peptone, soybean meal, peanut meal, fish meal, yeast extract, distillers' solubles and inorganic nitrates.  Other additions are (a) mineral salts such as sodium chloride, sodium sulphate, potassium salts, phosphates and trace metals; and (b) buffers, e.g. calcium carbonate.  B-mycin is isolated from the clarified broth by extraction with a water-immiscible solvent, e.g. butyl acetate or diethyl ether at a pH of 7.0 to 9.0.  The extract is concentrated and then extracted with an aqueous buffer of pH 1.0 to 4.0 and preferably 2.0, e.g. aqueous sodium phosphate of pH 2.0.  B-mycin is precipitated from the aqueous solution by adjusting the pH to 7.0 to 9.0.  Further purification by (a) repeating above solvent extraction on a crude; (b) adsorption on a magnesium silicate followed by elution with e.g. a mixture of chloroform and an alcohol such as ethanol, adsorption on a strongly acidic ion-exchanger such as "Amberlite" (Registered Trade Mark) 1R-120 followed by elution with 10 per cent aqueous sodium chloride may also be used; (c) forming salts with organic acids especially 3,5-dinitrobenzoic acid, salicylic acid, p-amino salicylic acid, and 3,5-dibromosalicylic acid. B-mycin is a white amorphous base; soluble in many organic solvents, e.g. alcohols, esters, chloroform, carbon tetrachloride, ethers, ketones, aromatic hydrocarbons; insoluble in aliphatic hydrocarbons, e.g. pentane; slightly soluble in water; stable in aqueous solution; melting point 143-155 DEG  C.; specific rotation [a ]20D = - 14.2; mol. wt. 770; probably C38H57-60N7 O7-8S; S = 59.58 per cent, H = 7.69 per cent, N = 12.79 per cent, S = 4.20 per cent, O = 15.74 per cent; ultra-violet absorption spectrum maximum at 2030   and shoulder at 2400  ; infra-red absorption spectrum in petroleum oil or hexachlorobutadiene shows bands at 3333, 2986, 1739, 1650, 1502, 1366, 1312, 1258, 1171, 1145, 1119, 976 and 806-800 cm-1.  Forms salts with (a) inorganic acids such as hydrochloric, sulphuric and phosphoric; and (b) organic acids such as acetic, phenylacetic, 3-nitrophthalic, anthranilic, acetylsalicylic, salicylic, p-aminosalicylic, benzoic, 3.5-dibromosalicylic, picric, 3.5-dibromo-4-aminosalicylic, 3.5 - dinitrosalicylic, 3.5 - dinitrobenzoic and penicillin.  B-mycin penicillinate is prepared by reacting penicillin with the free B-mycin, or a salt of penicillin with a salt of B-mycin, in an organic solvent.  Any form of penicillin, e.g. K, F, O or G, may be used.  B-mycin penicillinate has a prolonged action when injected (see Group VI).ALSO:A salt of penicillin with the antibiotic B-mycin, prepared by reacting penicillin with B-mycin, or a salt of penicillin with a salt of B-mycin (see Group IV(b)) when suspended in water and injected forms a preparation with prolonged action.