DESCRIPTIONS OF ANTIBIOTICS 



299 



Purification Ijy cliromatograpliy on alumina from 

 benzene. Elution with benzene, chloroform, and 

 chloroform-methanol (50:1) (active fraction). Re- 

 chromatographed on alumina and eluted with 

 chloroform. Subjected to countercurrent distribu- 

 tion (methanol-water-carbon tetrachloride-ehlo- 

 roform, 4:1:3:2). Water added to active fractions; 

 extracted with chloroform. E.xtract taken to dry- 

 ness. Residue extracted with ethyl acetate and 

 precipitated with ether. Recrystallization from 

 ethyl acetate-ether. 



Chemical and phi/sical properties: Neutral sub- 

 stance. Colorless thin plates. C24H.38O10: C = 

 59.27%; H = 7.94%; active H = 0.42%; O— CH;i = 

 6.52%; C— CH, = 9.38%. Ultraviolet absorption 

 s])ectrum maximum at 217 niyu (log e = 3.94). 

 Infrared absorption spectrum given in reference 

 1. Positive FeCl.-i te.st. Hydrogenation product has 

 no ultraviolet absorption spectnun maximum at 

 217 nxfi. May contain three to foin- C — CHg groups, 

 and an Q:,/3-unsaturated carl)onyl group. Forms a 

 monoacetyl derivative: colorless crystals; m.p. 

 218.5-220°C. 



Biological adivity: Very narrow r:inge of activ- 

 ity: B. megaieriian (0.1 to 1 jug per ml) and Strepto- 

 coccus pyogenes (10 ^g per ml). Other gram-positive 

 bacteria, including streptococci. Staph, aureus, 

 and mycobacteria, gram-negative l)acteria, and 

 yeasts are not affected. No activity on Strepto- 

 coccus pyogenes infection in mice. 



Toxicity: Mice tolerate 0.5 gm per kg sul)cutane- 

 ously. 



Reference: 1. Ettlinger, L. et at. Monatsh. 

 Cheni. J!«: 989-995, 1957. 



Melanoniyciii 



Produced by: Streptomyces utelanoyeues (2). 



Method of extraction: Broth-filtrate adjusted to 

 pH 4.0 and passed through a column of IRC-50 

 resin (H+ form) and the active fracticjn eluted 

 with 1 to 2 .V NH4OH. Addition of 10 per cent 

 acetic acid to pH 3.0 produces a l)lackish ])recipi- 

 tate. An aqueous solution of this material (pH 

 8.0 with 4 per cent NaOH) is dialyzed against tap 

 water overnight, then lyophilized. Another 

 method of extraction involves the use of IR4B 

 (Cl~ form) resin and elution with aqueous alkali. 

 The antibiotic can be precipitated with zinc 

 chloride, potassium alum, ammonium sulfate. 

 trichloroacetic, picric, and phosphotungstic acids, 

 ammonium reineckate, and other agents. Not 

 precipitated by methyl orange. 



Chemical and physical properties: Tasteless, 

 black-brownish, amorphous powder. Amphoteric. 

 Insoluble in acidic water and organic solvents. 



Soluble in water at i)H 6 to 9 and in methanol at 

 acid or alkaline pH. Soluble with inartivation in 

 acetone, l)utanol, and ethanol containing more 

 than 10 per cent alkaline water. Nondializal)le. 

 Positive xanthoproteic, diazo, sodium nitroprus- 

 side, and Millon tests. Acid hydrolysis jjroducts 

 include phenylalanine, leucine, proline, alanine, 

 arginine, histidine, glutamic acid, and glj'cine. 

 Ultraviolet absorption spectrum shows an indis- 

 tinct shoulder at 260 to 270 mfi. Isoelectric point 

 about pH 3.0. Na melanomycin: C = 54.74%; H = 

 6.99%; N = 9.87%; Na = 1.9%; Kjeldahl N = 

 9.00%. Activity decreased by 50 per cent after 

 heating to 100°C for 10 minutes. Hemolyzes horse 

 red blood cells at >124 yug per ml (1). 



Biological activity: Mild but definite effect 

 against Ehrlich carcinoma, lioth ascitic and sub- 

 cutaneous forms. No antibacterial or antifungal 

 activity (1). Anti-ascarid activity (ova) (3). 



Toxicity: Maximal tolerated dose = 250 mg per 

 kg subcutaneously, 50 mg per kg intravenously, 

 and 125 mg i)er kg intraperitoneally (1). 



References : 



1. Sugawara, R. et ul. J. Antibiotics (.Japan) 



1(»A: 133-137, 1957. 



2. Sugawara, R. and Onuma, .M. J. .\ntiliiotics 



(Japan) lOA: 138-142, 1957. 



3. Takaoka, M. et al. J. Antibiotics (Japan) 



llA: 134-137, 1958. 



Melaiiosporiii 



Produced by: Streptomyces mclanosporiis var. 

 melanosporofaciens. 



Remarks: The same organism jM-oduces elaio- 

 phylin. 



Method of extraction: Extraction of the mycelium 

 with n-butanol. Concentration of butanol to syrup. 

 Addition of ether to syrup yields a yellowish pre- 

 cipitate. White crystals of elaioj)hylin olitained 

 from the mother licpior, on standing. The precipi 

 tate contains melanosporin, elaioj^hylin, and other 

 antibiotics, including a polyene. Elaiophylin ex- 

 tracted with chloroform, followed by evaporation 

 and crystallization from acjueous ethanol. Melano- 

 sporin is left in the residue, which is suspeiuied in 

 anhydrous ethanol, stirred, and filtered, .\ddition 

 of ether to the ethanol precipitates impvu-e mela- 

 nosporin. Further purification by countercurrent 

 distril)Ution in cliloroform - methanol - w;it(M' 

 (2:2:1). 



Chemical and physical properties: \'ery light 

 yellow amorphous solid; m.p. 132-134°C. Soluble 

 in alcohols and dimethylformamide. Insoluble in 

 acetone, chloroform, and ether, [ajj," = +30° (c = 

 1.578 per cent in methanol). Light -absorption 



