Patent GB-757089-A (GB)
Title: Improvements in or relating to antibiotics
Filed: 1953-07-13
Abstract: A new antibiotic, antibiotic 106-7 is produced by cultivating Streptomyces garyphalus NRRL-2448, or a mutant thereof, in an aqueous nutrient medium under aerobic conditions of submerged or surface culture.  A temperature of 24 DEG  to 37 DEG  C., preferably 28 DEG C., a pH of 6 to 7 and duration of cultivation of 2 to 4 days is employed.  The medium contains (a) carbon source such as sugars, e.g. dextrose, sucrose, maltose, lactose and dextrin, and starches; (b) nitrogen source, e.g. casein hydrolysates, papaic digests of soybean meal, soybean meal, peanut meal, peanut oil meal, distiller's solubles, corn steep liquor, sodium nitrate, ammonium chloride and ammonium sulphate; (c) inorganic salts and trace metals.  Preferred is a medium containing 3 per cent soybean meal, 2 per cent dextrose, 0.75 per cent distiller's solubles and 0.25 per cent sodium chloride. A crude product is obtained by filtering the broth and drying the filtrate after, if desired, a treatment with charcoal.  The antibiotic may be recovered by adsorbing the antibiotic on a strongly acid cation-exchange resin on the alkaline cycle according to Specification 757,090 and eluting therefrom with an aqueous solution of a base.  The eluate is then adsorbed on to a strongly basic anion-exchange resin on the hydroxyl cycle, washed with water and eluted therefrom with an anion containing eluant.  The resins are strongly basic quaternary ammonium base-exchange resins, e.g. Amberlite IRA-400.  The eluant is (a) acid, e.g. hydrochloric, sulphuric or acetic at pH 4.5 to 7; (b) base, e.g. sodium hydroxide or potassium hydroxide at pH 7 to 14; or (c) salt, e.g. sodium chloride potassium chloride or sodium sulphate.  The crystalline base may be obtained by adjusting the pH of the eluate to 8.5, evaporating to small volume, adding a mixture of alcohols, e.g. ethyl alcohol and isopropanol and adjusting pH to 6.  It is soluble in water, insoluble in anhydrous alcohols, acetone, chloroform, ethyl acetate and pyridine; [a ]D23= +116 (water).  It is amphoteric forming (a) acid salts with e.g. hydrochloric and sulphuric acids; and (b) metal salts with alkali metals such as sodium, and alkaline earth metals such as calcium, barium and magnesium.  It is a wide spectrum antibiotic and contains the elements carbon, hydrogen, nitrogen and oxygen; decomposes at 150 DEG  to 155 DEG  C.; exhibits in aqueous solution at a pH of about 6, an ultra-violet absorption maxima at about 2260 ; exhibits in anhydrous crystalline form.  Absorption bands in the infra-red region of the spectrum when suspended in a hydrocarbon oil at the following frequencies expressed in microns: 2.8 to 4.4 (multiple bands), 4.65, 6.12, 6.20 (shoulder), 6.30, 6.42, 6.52, 7.10, 7.3, 7.5, 7.85, 8.15, 8.55 (shoulder), 8.80, 9.40, 10.68, 10.9, 11.2 to 11.3 (shoulder), 11.38, 12.08, 13.25 and 15.0.  The antibiotic may be used in pharmaceutical preparations (see Group VI).ALSO:Pharmaceutical and veterinary preparations for internal or external use comprise as an essential ingredient thereof the antibiotic 106-7 (see Group IV(b).  The antibiotic in the form of the base or salt, as the pure product or concentrate, may be employed in solution or in ointment base.