Patent GB-821819-A (GB)
Title: Production of an antibiotic designated 'novobiocin'
IPC: C07H17/075
Filed: 1956-04-18
Abstract: <PICT:0821819/IV (a)/1> A new antibiotic novobiocin, also known as cathomycin, is prepared by growing a novobiocin-producing-strain of Streptomyces spheroides, e.g. S. spheroides NRRL 2449 or mutants thereof, in an aqueous nutrient medium containing sources of carbon, nitrogen and inorganic salts under aerobic conditions.  The pH of the medium is 5.5 to 9.0 and preferably 7.0, the temperature of cultivation 20-37 DEG  C. and preferably 24-28 DEG  C. and the duration of cultivation 2 to 7 days.  Submerged cultivation is preferred. Sources of carbon are sugars, e.g. dextrose, sucrose, maltose, lactose and dextrin and starches.  Sources of nitrogen are casein hydrolysate, papaic digests of soybean meal, peanut meal, peanut oil meal, distillers' solubles, corn steep liquors, sodium nitrate, ammonium chloride and ammonium sulphate.  Sodium chloride may be the sole source of inorganic salts apart from trace metals. Novobiocin may be extracted from an alkaline clarified broth by means of butanol, or by acid precipitation as in Specification 821,820. Further purification is by adsorption on Specification 821,820 by alumina chromatography or treatment with petroleum ether to remove impurities.  The procedures may be combined.  Novobiocin forms salts with inorganic and organic bases.  Examples relate to the preparation of the following salts: (a) monosodium novobiocin; (b) calcium novobiocin; (c) magnesium novobiocin; (d) novobiocin-spiramycin; (e) novobiocin neomycin; (f) novobiocin streptomycin; (g) novobiocin dihydrostreptomycin. Novobiocin also forms salts with benzyl trimethylammonium acid salts, quinine, procaine, guanidine, N-benzyl-b -phenylethylamine, noformicin, cyclohexylamine, N-ethylpiperidine, triethylamine, dicyclohexylamine, ethylenediamine, isobutylamine, arginine, histidine and lysine.  Novobiocin is acidic, having two base binding groups, it is soluble in aqueous alkaline solutions, in methanol, butanol, ethanol, ethyl acetate, acetic acid, dioxane, acetone and methyl ethyl ketone, insoluble in ether, benzene, chloroform, carbon tetrachloride, ethylene dichloride, water and hydrochloric acid; it may be obtained in two crystalline forms; it is optically active [a ]25D = - 27 DEG  in 1N sodium hydroxide; in 0.1N sodium hydroxide exhibits a characteristic ultra - violet absorption peak at 307 mu (E1%1 cm. 600); in the infra-red it shows characteristic peaks at 5.8-6.0, 6.10, 6.21, 6.30, 6.49, 6.63, 7.4-7.6, 7.78, 7.96, 8.27, 8.60, 8.70, 9.13, 9.40, 10.0-10.1, 10.28, 10.60, 12.0-12.30, 12.60-12.75, 13.07 and 13.39 microns; it contains 60.24% carbon, 6.56% hydrogen, 4.86% nitrogen and 29.3% oxygen; possibly C30-32-H34-42N2O10-12; molecular weight 592\sB25. Novobiocin is active against gram positive bacteria. It may be used as a general disinfectant, as an animal medicine (see Group VI), as a medicine for respiratory diseases (see Group VI), and as a preparation for the treatment of plant diseases (see Group VI). Specification 791,731 also is referred to.ALSO:The antibiotic novobiocin (cathomycin) is incorporated into ointments for the treatment of mastitis in cows, or put up in the form of capsules or tablets for the treatment of respiratory diseases in humans.  A tablet contains sodium novobiocin granulated with cellulose acetate hydrogen phthalate in methanol acetone and compressed into tablets. Aqueous sprays containing sodium novobiocin, wetting or spreading agents and/or other active agents may be used for the treatment and control of plant diseases.  Specifications 791,731 and 821,820, [Group IV (b)], are referred to.