Patent GB-738537-A (GB)
Title: Improvements in or relating to antibiotics
Filed: 1953-07-14
Abstract: A new antibiotic, carbomycin, is produced by cultivating Streptomyces sp. Waksman Collection 3628 (New Jersey Agricultural Experiment Station) under aerobic conditions of submerged or surface culture in an aqueous nutrient medium containing assimilable sources of carbon and nitrogen.  The pH is 5 to 9, and preferably 7.0; the temperature 20 DEG  to 40 DEG  C., and preferably 25 DEG  C.  The carbon source is starch, dextrose, sucrose, maltose, glycerol; a fat, e.g. a vegetable oil such as cottonseed oil, an animal fat such as mutton tallow, or a triglyceride such as tristearin; and a fatty acid such as acetic acid or palmitic acid. The nitrogen source may be organic, e.g. soybean meal, corn steep liquor, meat extract, casein, fish meal, liver cake or distillers' solubles; or inorganic, e.g. nitrates and ammonium salts. Optional additions are (a) precursors, e.g. cobalt salts, progesterone or 11-desoxy-17-hydroxycorticosterone; (b) buffers, e.g. CaCO3 and (c) mineral salts of metals such as potassium, calcium, magnesium, sulphur and iron; phosphates.  The antibiotic is recovered from the filtered fermentation broth by solvent extraction with a water-immiscible organic solvent such as benzene, chloroform, amyl acetate, ethyl acetate, butanol, amyl alcohol, trichlorethylene and ether at a pH of 6.6.  The water-soluble fraction obtained by distilling off the organic solvent in the presence of water is freeze dried and the antibiotic recovered by crystallization of the solid from an organic solvent such as isopropanol.  Alternatively the solid may be dissolved in 10 per cent acetone in benzene, passed down a column of silica gel, eluted therefrom with 30 per cent acetone-benzene and the antibiotic recovered as above.  The antibiotic base contains 59,24 per cent C, 8.64 per cent H, 1.76 per cent N and 7.33 per cent methoxyl (C42H73NO16); it melts at 207-209 DEG  C., [a ]22D = -57 DEG  \sB 3 DEG  (ethanol), [a ]22D = -53 DEG  \sB 3D (0.05N acetic); gives a precipitate with phosphotungstic acid, picric acid, methyl orange and tetrathiocyanodiammonochromic acid; reduces potassium permanganate; in 2 per cent sodium dihydrogen phosphate the ultra-violet absorption curve shows a maximum at 1% 241 mm , E1 cm. = 158, in methanol maximum 1% is at 240 mm , E1 cm. = 157; in mineral oil absorption in the infra-red occurs at 2899, 1736, 1701, 1637, 1464, 1427, 1381, 1305, 1271, 1238, 1193, 1160, 1129, 1110, 1095, 1075, 1060, 1034, 1011, 989, 962, 944, 928, 913, 873, 861, 840, 821, 805, 781, 742 and 689 cm.; Rf values for different systems are given.  Specified salts are the hydrochloride sulphate, phosphate, citrate, tartrate, gluconate and p-toluene sulphonate.  The antibiotic is employed in pharmaceutical and veterinary compositions (see Group VI).ALSO:An antibiotic, carbomycin, produced by cultivating streptomyces sp. Waksman Collection 3628 (see Group IV (b)) is an essential ingredient of pharmaceutical and veterinary preparations of oral, parenteral or topical form.  Specified are oil solutions and admixtures with other antibiotics such as neomycin or chemotherapeutic agents such as N-hydroxy-2-pyridinethione.