Patent GB-923061-A (GB)
Title: The antibiotic holothin, its acyl derivatives and hydrohalides and pharmaceutical preparations containing such compounds
Filed: 1959-07-27
Abstract: A new antibiotic Holomycin of formula:  <FORM:0923061/IV(a)/1> where R = .-CO.CH3 is produced by cultivating a Holomycin producing strain of streptomyces griseus especially S. griseus 2764 under aerobic conditions in an aqueous nutrient medium containing one or more inorganic salts, a source of carbon and a source of nitrogen at 18-40 DEG C.  Specified inorganic salts are chlorides, nitrates, carbonates or sulphates of alkali or alkaline earth metals, iron, zinc or manganese.  Specified carbon sources are carbohydrates such as glucose, saccharose, lactose or starch.  Specified nitrogen sources are amino acids and mixtures thereof, peptides and proteins and their hydrolysates such as peptone or tryptone, meat extracts, water soluble constituents of grain such as maize or wheat, of distillation residues in the manufacture of alcohol, of yeast, of beans especially soyabeans and of seeds, e.g. cottonseed.  Submerged aerobic cultivation at 35 DEG  to 37 DEG C. for 15 to 72 hours is preferred. The antibiotic is isolated from the clarified broth (a) by solvent extraction with a water immiscible organic solvent, e.g. ethyl acetate, chloroform, butanol or diethyl ether (b) absorption on active carbon or an activated earth, e.g. Fuller's earth or "Floridin" (Registered Trade Mark) and elution with at least partially water miscible organic solvent, e.g. acetone, butanol or methylethyl ketone. Impurities are removed by extraction with first aqueous acid pH\sE5 and then with aqueous alkali pH\sF8.  On evaporation in vacuo the crude Holomycin is obtained as a brown oil. The crude Holomycin is further purified by (a) chromatography using alumina, silica gel or magnesium silicate as absorbent (b) distribution between an aqueous solution and a water immiscible solvent particularly by the counter current method using as solvent system ethyl acetate and water (c) recrystallisation (d) sublimation. On acid hydrolysis Holomycin gives a product Holothin of formula <FORM:0923061/IV(a)/2> which on acylation with a fatty acid chloride or anhydride yields acyl derivatives, e.g. the lower alkanoyl such as butyryl and propionyl and phenyl lower alkanoyl derivatives, and with hydrochloric acid gives holothin hydrochloride.  When treated with hydrogen and Raney Nickel Holomycin gives a product of formula:- <FORM:0923061/IV(a)/3> which on acid hydrolysis gives a :g  diamiono valeric acid.  The new antibiotics have the following properties:- <TABS:0923061/IV(a)/1> The antibiotics Holomycin and Holothin and its acyl derivatives and hydrohalides or mixtures thereof are used in medicinal preparations (see Group VI).ALSO:A pharmaceutical preparation for enteral, parenteral or local administration comprises Holomycin or Holothin or one of its acyl derivatives or hydrohalides or mixtures thereof (see Group IV (b)) in admixture with an organic or inorganic carrier.  The preparation may be in the form of a tablet, dragee, powder, salve, cream, suppository, solution, suspension or emulsion and may also contain preserving, stabilizing, wetting or emulsifying agents. Specified carriers are gelatin lactose, starches, magnesium stearate talc, vegetable oils, benzyl alcohols, gums; polyalkylene glycols, white petroleum jelly or cholesterol.