DESCRIPTIONS OF ANTIBIOTICS 



269 



in mice against exjioriniental infections caused 

 bv B. ilniHdlitidls and ('ryptorocfiis )ieofannaiis 



(1,3). 



Toxicity: LDju (mice) 3 mg per kg intravenously, 

 17 mg per kg intraperitoneally, 12 mg per kg intra- 

 muscularly, and 40 mg per kg suhcutaneously (1). 



References: 



1. Charney, J.c/a/. Antibiotics Ami. 228-235, 



1955-1956. 



2. Muller, W. H. Am. J. Botany 45: 183-190, 



1958. 



3. Solotorovsky, M. cl nl . Antil)iotics & Chem- 



otherapy 8: 3(14-371, 1958. 



4. Harmon, R. E. ct al. J. Am. Chem. Soc. 



80: 5173-5178, 1958. 



Kiim\ oelin 



Produced by: Strcptoniyces sp. 



Method of extraction: Washed mycelium ex- 

 tracted with methanol, the extract concentrated 

 in vcK IK), and lyophilized. Residue extracted re- 

 peatedly with ethanol, and the ethanol extract 

 concentrated. Baryta water is athled to the con- 

 centrate until no more ])reci])itat ion occurs. Pre- 

 cipitate removed l)y centrit'ugation; excess 

 Ba(OH)2 precipitated l)y CO2 . An inactive i)re- 

 cipitate forms on adding an etiual volume of H2O, 

 and is filtered off. Eumycetin ])recipitated on 

 chilling for 24 to 48 hours. Recrystallized from 

 50 per cent ethanol. 



Chemical and phi/t^ical properties: Colorless 

 needles; m.p. 148-150°C. Soluble in methanol, 

 ethanol, l)utanol, acetone, ether, chloroform, and 

 ethyl acetate. Sparingly soluble or insoluble in 

 water, 10 per cent HCl, and 10 per cent NaOH. 

 Negative biuret, Fehling, ninhydrin, Alillon, 

 Molisch, Liebermann-Burchard, Sakaguchi, and 

 Rosenheim tests. Positive FeCls and diazo tests. 

 No color produced by addition of concentrated 

 sulfuric or hydrochloric acids. Ultraviolet spec- 

 trum shows peak at 302 ni/i (c = 0.02 per cent in 

 methanol). 



Biological activifi/: Active on fungi. Inhibits 

 after 72 hours: Tonila rubra, M jicodernia sp., and 

 P. chrysogenum at <0.05 pg per ml; Oidiuni luctis 

 at 1 /ug per ml; Aspergillus oryzae at 0.325 ^tg per 

 ml; Willia anomala at 0.5 jug per ml; Saccharoniy- 

 ces sake at 1.0 ng per ml; C. albicans at 10 fxg per 

 ml; and Trichophyton interdigitale at 0.15 /ig per 

 ml. Active against Xocardia and Streptoniyccs. 

 Not active against bacteria. Mycobacterium (107 

 inhibited by 100 /ug per ml. Activity against ('. 

 albicans affected by horse serum. No hemolytic 

 activity against rabbit red blood cells. 



Toxicity: LDso (iu water with carboxymethyl- 



cellulose) 3.0 mg per kg svd)cutaneously in mice, 

 2.2 mg per kg intraperitoneally. 



Reference: 1. Aral, T. and Takamizawa, Y. J. 

 Antibiotics (Japan) 7A: lt)5-l(J8, 1954. 



Eiiriniyciii 



Produced by: Streptoniyccs sp. 



Method of extraction: Clarification of broth at 

 pH 2.0 with 1 per cent "Kerozite" with agitation 

 for 1 hour. Active filtrate adjusted to pH 8.2 with 

 sodium hydroxide; the precipitate that forms is 

 filtered off. The filtrate treated with 1.5 per cent 

 Kerozite (2 hovu'S of agitation). Elution with acidic 

 methanol (0.37 per cent HCl). Neutralization of 

 the eluate with sodium hydroxide, and concentra- 

 tion in vacuo. Addition of acetone in excess gives 

 an inactive precipitate, and the antibiotic is pre- 

 cipitated as the picrate from the active layer. The 

 solid hydrochloride may be precipitated by pe- 

 troleum ether. Purification by chromatography 

 on alumina. Inactive impurities removed with 

 acidic methanol (10 p(>r cent formic acid) and the 

 antibiotic eluted with water. 



Chemical and physical properties: Paper chroma- 

 tography (n-butanol -propionic acid-water, 4:1:5) 

 indicates three constituents, of which two are ac- 

 tive and ninhydrin-positive. Light cream-colored. 

 Sulfate very solul)le in water (hygroscopic), solu- 

 ble in methanol, slightly solul^le in ethanol, and 

 very slightly soluble in acetone. Practically in- 

 soluble in ether and ligroin. Precipitated as a pic- 

 rate, phosphotungstate, helianthate, and reineck- 

 ate. Positive ninhydrin, Sakaguchi, biuret, and 

 creatinine tests. Creatinine test more strongly 

 positive after acid hydrolysis. Ammonia is released 

 during alkaline hydrolysis, with loss of a guanidino 

 group. Negative maltol, Schifl', Fehling, and glu- 

 cosamine tests. A salmon color in 3.5 per cent HCl 

 is intensified by boiling; a pale yellow color in 10 

 per cent NaHCO.-j fades on boiling. Thermostable. 

 Little loss in activity noted after heating for 3 

 hotu's in 5 per cent BaOH over a steam bath, or 

 30 minutes at 100°C at pH 2.0. The principal sub- 

 stance, purified by chromatography on a cellulose 

 column and developed with n-butanol-acetic acid- 

 water (4:1.8:5), decomjjoses at 155°C (chloride) 

 andl30°C (sulfate). 



Biological activity: Active on gram-positive and 

 gram-negative bacteria and mycol)acteria. Pro- 

 tects mice and partially protects rabbits against 

 infection with Staph, aureus, (iuinea pigs pro- 

 tected against infection with B. anthraci.'^. 



Toxicity: LDo (mice) 2(X) mg per kg sul)cutane- 

 ously; 700 mg per kg (rabbits); and 300 mg per 

 kg (guinea pigs). 



