Patent GB-768971-A (GB)
Title: Antibiotic d-52, its salts and method of preparation
Filed: 1955-04-07
Abstract: <PICT:0768971/IV(b)/1> <PICT:0768971/IV(b)/2> <PICT:0768971/IV(b)/3> A new antibiotic, designated antibiotic D-52, is produced by cultivating under aerobic conditions an antibiotic D-52-producing strain of Streptomyces caelestis, especially Streptomyces caelestis NRRL 2418, in a culture medium containing assimilable sources of carbohydrate, nitrogen and inorganic salts.  Specified sources of (a) carbohydrates, are brown sugar, lactose, dextrin, starch, sucrose, glucose and molasses; (b) nitrogen, are corn steep liquor, brewers' yeast, distillers' solutes, soyabean meal or flour, casein, amino acid mixtures, peptones, nitrates and ammonium sulphate; (c) inorganic salts, are salts yielding sodium, potassium, calcium, phosphate, chloride and sulphate ions. Essential trace elements also required.  Culture may be surface or submerged.  The initial pH of the medium is 6.5 to 7.5 and preferably 7.0.  The temperature of cultivation 20 DEG  to 32 DEG  C., and preferably 24 DEG  to 28 DEG C., and the duration of cultivation is 2 to 6 days (submerged) or 4 to 8 days (surface).  Antibiotic D-52 is isolated from the clarified broth by (a) solvent extraction or (b) adsorption.  In the solvent extraction, the broth is adjusted to pH 6 to 10 and preferably 7.5 to 8.0 and extracted with a water-immiscible polar solvent, e.g. chlorinated hydrocarbon such as chloroform, an alcohol such as butanol, an alkyl ester such as ethyl acetate, or a ketone such as methyl isobutyl ketone.  The extract is evaporated in vacuo to produce the crude antibiotic. Alternatively, the extract is concentrated and a non-solvent, e.g. hexane, added to precipitate the antibiotic.  Adsorbing agents used in the adsorption process are activated carbon especially when pretreated with acetic acid, activated alumina, silica gel, and magnesium aluminium silicate. Elution is by a polar organic solvent for the antibiotic.  Antibiotic D-52 has a broad antibacterial spectrum.  It is amphoteric, being soluble in water at a pH 1 to 7, insoluble at pH 7.5 to 9.0 and soluble at pH 10 to 13.  It is stable at pH 2 to 7.  It is soluble in methanol, chloroform, ethyl acetate and methylene chloride but insoluble in ether and ligroin.  Empirical formula C23H36-40O9N2S.[d]24D = 121.5 DEG  (in chloroform).  Ultraviolet absorption in aqueous solution gives a maxima of E1 per cent1 cm.=182 at 239 millimicrons and E1 per cent1 cm.=74 at 307 millimicrons.  A suspension in liquid petrolatum exhibits characteristic absorption in infra-red at 3340, 3210, 1900, 1672, 1655, 1570, 1545, 1488, 1090 and 758 cm.-1.  Examples describe the preparation of (a) antibiotic D-52 hydrochloride by passing dry hydrochloric acid into a methylene chloride solution of D-52; (b) D-52-oxalate by adding oxalic acid dihydrate to a methanol solution of D-52 and adding ether to precipitate the oxalate; (c) D-52 salicylate by adding salicylic acid to a methanol solution of D-52 and adding ether to precipitate the salicylate.  Antibiotic D-52 is incorporated into preparations for the treatment of man, animals and plants (see Group VI).ALSO:A new antibiotic D52 (see Group IV(b)) is used as a feed supplement in promoting the growth of animal and poultry alone or in conjunction with one or more of the following antibiotics: tetracycline, chlortetracycline, oxytetracycline, streptomycin, dihydrostreptomycin, penicillin, chloramphenicol, bacitracin, actidione, circulin, tyrothricin, polymixin B, endomycin, amicetin, fumagillin and erythromycin.ALSO:A new antibiotic D-52 is incorporated into preparations for the treatment of man, animal and plants.  In the case of plants, it is used alone or in admixture with one or more of the antibiotics oxytetracycline, chlortetracycline, penicillen, streptomycin or actidione.  In the case of man and animals, antibiotic D-52 may be combined with one or more of the following: (a) antibiotics such as streptomycin, dihydrostreptomycin oxytetracycline, tetracycline, chlortetracycline, bacitracin, circulin, penicillins, e.g. penicillin O procaine penicillin and chloroprocaine penicillin, chloramphenicol, tyrothricin, polymixin B, endomycin, amicetin, fumagillin and erythromycin; (b) sulpha compounds such as sulphadiazine, sulphamerazine and sulphamethazine; (c) steroid hormones such as cortisone, hydrocortisone, and esters thereof; halo-cortisones such as 9-a -chlorocortisones, and 9 - a  - fluoro-cortisone, and esters thereof; (d) estrogens; (e) analgines, e.g. salicylates; (f) antihistaminies such as pyrrolidine ethyl phenothiazine hydrochloride and N1-pyridyl-N1-benzyl-N-dimethyl ethylene diamine; and (g) antifungal agents such as caprylic acid, undecylenic acid and p-hydroxybenzoic acid.