398 



DESCRIPTIONS OF ANTIBIOTICS 



with 0.067 M (pH 8.0) Sorensen's pho.sphate buffer 

 gives Fraction A. B can be eluted with 20 per cent 

 pyridine-5 per cent HCl (95:5), and C with pyri- 

 dine-butanol-water (3:4:7). Fraction A precipi- 

 tates as the 2,5-diamino-7-ethoxvacridine (riva- 

 nol) salt. B is purified by adding 20 per cent 

 ammonium sulfate solution-butanol (25:5) to 

 active fraction to force it into the l)utanol layer. 

 B precipitates on addition of benzene to butanol. 

 Fraction C concentrated, and 20 per cent am- 

 monium sulfate added. Lower layer lyophilized to 

 give C (15). Fractions further purified by counter- 

 current distribution (chloroform-methanol-pH 

 8.4 borate buffer, 2:2:1) to give pure A and B (24). 

 Chemical and physical properties: Complex com- 

 posed of three biologically active components, A, 

 B, and C. Acidic conjugated heptaenes, possibly 

 containing a lactone group. Complex: Yellow sub- 

 stance. Decomposes at about 155°C, turning brick - 

 red. Cannot be sublimed. Very soluble in propy- 

 lene glycol, benzyl alcohol, pyridine, acetic acid, 

 phenol, and the following when water is present: 

 l)utanol, ethylene glycol monomethyl ether, di- 

 oxane, propanol, and methyl ethyl ketone. Moder- 

 ately soluble when water is present in acetone, 

 methanol, and ethanol. Very slightly soluble in 

 water. Insoluble in xylene, ether, ethyl acetate, 

 benzene, toluene, carbon disulfide, petroleum 

 ether, ligroin, chloroform, and CCI4 . Most soluble 

 at alkaline pH. Ultraviolet absorption spectrum 

 maxima at 286, 346, 364, 384, and 405 ni/x (ethanol). 

 These peaks are flattened to a plateau at 320 to 

 350 niM or to two peaks at 235 and 335 m/j, in water. 

 Infrared data given in reference 5. Positive diazo, 

 carbylamine, and Ehrlich tests. Negative ninhy- 

 drin, biuret, Fehling, Benedict, Molisch, Tollen, 

 and quinone tests. Blue color in concentrated 

 HCl or H2SO4 turns purple on standing. Most 

 stable at pH 6 to 7. Heat- and photo-labile. De- 

 stroyed by ultraviolet light. Precipitated by bi- 

 valent metals, procaine, dihydrostreptomycin, 

 and acrinol. No S or halogens. Conflicting reports 

 on presence of N (see under components below). 

 Ozonol,ysis products include acetone, glyoxal, and 

 p-acetaminobenzoic acid. Oxidation with KMn04 

 .■yields p-aminoacetophenone and oxalic acid (1, 2, 

 5-7, 16-18, 22-24). Components A and B: Light 

 yellow amorphous powders which turn deep yellow 

 at 150-100°C and brown-yellow at 180°C. Do not 

 melt or decompose when heated to 320°C. Soluble 

 in glacial acetic acid, pyridine, dimethylform- 

 amide, and alkaline water. Insoluble in petroleum 

 ether, ether, benzene, chloroform, and carbon 

 disulfide. Positive Ehrlich (yellow), Ehrlich- 

 Harter (red), diazo (red), FeCl, (pale violet). 



Carr-Price (blue-violet), Bayer, bromine, carbyl- 

 amine, and dithiocarbamate tests. Uncertain re- 

 actions with 2,4-dinitrophenylhydrazine, Tollen, 

 and Molisch tests. Contain C, H, O, and N. Nega- 

 tive ninhydrin, Fehling, and Benedict tests. Ultra- 

 violet absorption spectrum maxima at 235, 340, 

 358, 377, and 400 m/j.. Infrared spectrum given in 

 reference 24. Fraction B in dilute aqueous alkali 

 forms a precipitate at pH 9.0; A at pH 10.0 (24). 



Biological activity: Active on Trichomonas, 

 Endamoeha histolytica, Treponema pallidum, and 

 Trypanosoma cruzi. Active on yeasts, fungi, and 

 anaerobic bacteria, but not on aerobic bacteria. 

 Activity reduced by serum. Resistance to tricho- 

 mycin accompanied (in Candida) by decreased 

 virulence for mice (1-3, 5, 9, 13, 16). /n vivo: Active 

 (mice) on Trichomonas vaginalis, C. albicans, 

 Treponema pallidum (rabbits), Endamoeba histo- 

 lytica (mice), and Trichophyton asteroides (guinea 

 pigs) (3, 5, 8, 20, 21). Prolongs survival time in 

 mice infected with Cryptococcus neoformans (25). 



Toxicity: LD.50 (mice) 2.2 mg per kg intraperi- 

 toneally and intravenously, and 17 mg per kg 

 subcutaneously (12). Mice tolerate 300 mg per kg 

 orally, but not 1 gm per kg (4). 



Utilization: Vaginal candidiasis. Trichomonas 

 vaginalis infections (3, 5). Otitis media caused by 

 Candida spp. or Absidia ramnsa (11). Trichophy- 

 tosis (8). 



References: 



1. Hosoya, S. et al. J. Antibiotics (Japan) 



5: 564-566, 1952. 



2. Hosoya, S. et al. Japan. J. Exptl. Med. 



22: 505-509, 1952. 



3. Hosoya, S. et al. J. Antibiotics (Japan) 



6A: 92-97, 1953. 



4. Hosoya, S. et al. J. Antibiotics ("Japan) 



6A: 98-100, 1953. 



5. Hosoya, S. Chemotherapy 2: 1-14, 1954. 



6. Vining, L. C. et al. Congr. intern. Botan. 



Vol. prelim. 8, Sec. 24: 106-110, 1954. 



7. Hosoya, S. et al. Intern. Med. News 14: 



1-2, 1954. 



8. Hosoya, S. et al. Giorn. ital. chemioterap. 



1: 217-230, 1954. 



9. Magara, M. et al. Antibiotics & Chemo- 



therapy 4: 4:33-438, 1954. 



10. Yamaguchi, T. J. Antibiotics (Japan) 



7A: 10-14, 1954. 



11. Sambe, B. et al. J. Antiliiotics (Japan) 



7A: 132-133, 1954. 



12. Ozaki, M. et al. J. Antil)iotics (Japan) 



7A: 159-164, 1954. 



13. Dimmling, T. Zentr. Bakteriol. Para- 



sitenk., Orig. 163: 530-538, 1955. 



