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A Streptomyces strain which is similar to S. diastatochromogenes (Krainsky) Waksman et Henrici 1948 sensu Hütter (1967) was found to produce a new antibiotic designated trypanomycin. The red-pigment antibiotic, which has novel trypanocidal activity in vitro and in vivo, was isolated from a C-, N-, and iron-containing culture of the strain IMET JA 10081/9 by extraction with organic solvents, transfer into the aqueous phase, reextraction with organic solvents at p H 6.8, precipitation by hydrocarbons, and purification by chromatographic methods. Trypanomycin has indicator properties. The main constituents of the antibiotic mixture are readily soluble in water and are very stable in distilled water at room temperature (28 C) for 24 hr. The composition of the base of trypanomycin A 2 (melting point, 175 to 183 C) corresponds to the empirical formula C 41–42 H 52-56 NO 21-22 . The absorption spectra in the ultraviolet and visible regions are very similar to those of the 4,5,8-trioxyanthrachinones. Trypanomycin has strong antiprotozoal activity, e.g., against trypanosomes. The natural substance additionally inhibits the growth of gram-positive bacteria, stable as well as unstable L-forms of gram-negative bacteria, mycoplasmas, yeast protoplasts, and tumor cells in vitro. The LD 50 of trypanomycin in mice was 60 mg/kg when administered intravenously and 31 mg/kg on intraperitoneal administration. If the antibiotic was added to cultures of animal or human cells in vitro, mitotic inhibition and chromosomal aberrations resulted. Trypanomycin differs in its biological activities and chromatographic behavior from other anthracyclines, e.g., cinerubine A and B, daunomycin, adriamycin, nogalamycin, rutilantin, pyrromycin, cyclamycin, and ryemycin.