Patent GB-1064300-A (GB)
Title: A new antibiotic coumermycin
IPC: C07H17/075
Filed: 1962-12-17
Abstract: The invention relates to the antibiotic coumermycin (Notomycin, Registered Trade Mark) which is an acidic substance, effective in inhibiting the growth of Gram positive bacteria, which is readily soluble in acetone, <PICT:1064300/C2/1> <PICT:1064300/C2/2> dioxane and alkaline water, moderately soluble in benzene, methanol and chloroform and insoluble in carbon tetrachloride, petroleum ether and acidic water, which gives positive Fehling and Molisch reactions, decolorizes bromine, and gives negative ninhydrin, Tollens and anthrone reactions, and which in a purified form melts at 222-224 DEG C., exhibits <FORM:1064300/C2/1> of - 134 DEG  (C = 1.0% acetone), exhibits an ultraviolet absorption spectrum (Fig. 2) in ethanol having maxima at 275 mm  <FORM:1064300/C2/2> , 335 mm <FORM:1064300/C2/3> , in N/10 HCl having maxima at 275 mm <FORM:1064300/C2/4> and 345 mm  <FORM:1064300/C2/5> , has a neutral equivalent of 548, an elementary analysis C = 59.1%, H = 5.35%, N = 5.90% and O = 29.65%; and exhibits absorption in the infra-red as shown in Fig. 1. Coumermycin forms salts with (a) aqueous solutions of alkali metal and alkaline earth metal hydroxides, e.g. sodium and calcium coumermycin, (b) organic bases, e.g. procaine, N - benzyl - b - phenethylamine, N,N1 - bis(dehydroabiety) ethylene diamine, ephenamine, N,N1 - dibenzylethylene diamine, dehydroabietylamine, dicyclohexylamine, streptomycin and dihydrostreptomycin. Coumermycin may be extracted from the whole or clarified broth at acid pH, e.g. pH 6.0 by means of a waterimmiscible solvent, e.g. methyl isobutyl ketone, the extract treated with water of pH 10.0 and the aqueous extract so-formed treated with a water - immiscible solvent.  After concentration, coumermycin is precipitated from the organic concentrate by addition of a nonsolvent such as petroleum ether. Further purification may be by (a) chromatography on acid treated alumina followed by elution with methanol; (b) countercurrent distribution using carbon tetrachloride/chloroform/methanol/water 5 : 1 : 5 : 1 ; and (c) recrystallization from aqueous acetone and methanol.  The antibiotic may be obtained from separated mycelium by extraction with acetone, removal of acetone and reextraction of aqueous residue with methyl isobutyl ketone.  Coumermycin is produced by cultivating a coumermycin-producing strain of Streptomyces rishiriensis nov. spec., e.g. ATCC 14812 in an aqueous nutrient medium under submerged aerobic conditions.  A suitable medium contains cottonseed endosperm flour, soluble starch, yeast extract, dibasic potassium phosphate, sodium chloride, calcium chloride, magnesium sulphate, zinc sulphate and water.ALSO:A pharmaceutical composition comprises an antibiotic coumermycin (Notomycin, registered Trade Mark) and a carrier. Mastitis in cattle and scours in calves may be treated by a vegetable oil suspension of the antibiotic or capsules.  Optional ingredients are (a) antihistamines, (b) sulpha drugs e.g. sulphadiazine, (c) lipotropic agents e.g. methionine, choline, inositol and betasitosterol, (d) central nervous system stimulants e.g. caffeine and ampheta -mines, (e) local anaesthetics, (f) analgesics e.g. aspirin, salicylamide, sodium gentisate, codiene, p-acetylaminophenol and phenacetin, (g) sedatives e.g. barbiturates and bromides, (h) antibiotics e.g. penicillin, streptomycin, bactiracin, polymyxin, tyrothricia, erythromycin, tetracyclines, oleandomycin, neomycin, chloramphenicol, novobiocin, cycloserine, kanamycin and carbomycin, (i) vitamins, (j) hormones e.g. cortisone, 9-a -fluorocortisone, prednisone and prednisolone, (k) anabolic agents e.g. 11,17-dihydroxy-9-a -fluoro-17-a mehyl-4-androsten-3-one, and antifungal agents e.g. mycostatin.