DESCRIPTIONS OF ANTIBIOTICS 



217 



2. Soluble in methanol, ethanol, and chloroform. 

 Moderatel}^ soluble in acetone and ethyl acetate. 

 Slightly soluble in ether and benzene. Insoluble in 

 water and petroleum ether. Dark brownish color 

 in Tollen reaction, with no mirror. Green color in 

 Fehling test. Permanganate solution decolorized. 

 Molisch, anthrone, and FeCU tests negative. 

 Most stable at an acid pH (2). Azalomycin F : 

 Neutral compound. White needle-shaped crystals; 

 m.p. 125-127°C (decomposition). C = 60.41%; 

 H = 8.57%; N = 4.33%. C3oH.=,oOioN2 . Molecular 

 weight (Berger-Akiya) 600. Molecular weight 

 598.72. [q;]jj = +46 (c = 1 per cent in methanol). 

 Light-absorption maxima at 240 and 268 m^ in 

 methanol. Infrared absorption spectrum given in 

 reference 2. Soluble in methanol and ethanol. 

 Moderately soluble in 20 per cent aqueous acetone. 

 Slightly soluble in acidic water. Insoluble in alka- 

 line water, acetone, ethyl acetate, and chloroform. 

 Dark brown color in Tollen reaction. Brown color 

 in concentrated H2SO4 . Wine-color in concen- 

 trated HCl. Positive Molisch, anthrone, Fehling, 

 FeCls , ninhj'drin, Millon, and biuret tests. Posi- 

 tive ninhydrin reaction after 2 minutes of hy- 

 drolvsis with 5 .V HCl. Most stable at an alkaline 

 pH.' 



Biological activity: Azaloinijcin B: Active in 

 ritro against gram-positive bacteria, including 

 Clostridia. Inactive against mvcobacteria, gram- 

 negative bacteria, and fungi. Azalomycin F: Active 

 in vitro against gram-positive bacteria and fungi, 

 including yeasts and Trichomonas vaginalis. 



Toxicity: LD50 (mice): azalomycin B, 281 mg 

 per kg; azalomycin F, 25.9 mg per kg intraperi- 

 toneally. 



References: 



1. Arai, M. J. Antibiotics (Japan) 13A: 



46-50, 19(J0. 



2. Arai, M. J. Antibiotics (Japan) 13A: 



51-56, 1960. 



Azaserine 



Produced by: Streptomyces fragilis (1). One or 

 more antibiotics in addition to azaserine are pro- 

 duced In' this culture (12). 



Method of extraction: I. Broth-filtrates flash- 

 evaporated at <35°C. Concentrates extracted into 

 95 per cent ethanol and extracts filtered; chroma- 

 tographed on alumina at pH 5.0 to 6.0 or pH 7 to 

 8 and developed with graded quantities of water 

 in ethanol and finally water. Active eluates are 

 concentrated, adsorbed on carbon from 1 to 2 per 

 cent aciueous acetone or from water, and eluted 

 with 1 to 5 per cent acetone. Active fractions are 

 freeze dried. Crj'stallized from boiling 90 ])er cent 



ethanol on cooling. Recrystallized from 90 per 

 cent methanol or 90 per cent ethanol (4). 



Chemical and physical properties: O-Diazoace- 

 tyl-L-serine (5). Long light yellow-green needles; 

 m.p. 146-162°C (decomposition). Verj^ soluble in 

 water. Slight solubility in cold absolute methanol, 

 ethanol, and acetone increased by warming. Ultra- 

 violet absorption spectrum maximum at 250.5 m/x 

 (E\7m 1140) (pH 7.0 phosphate buffer). In 0.1 A' 

 NaOH, biological activity is destroyed and the 

 ultraviolet absorption maximum shifts to 252 m/n 

 {E\!'^ 1230). In 0.1 .V HCl, No is evolved, biologi- 

 cal activity is destroyed, and the ultraviolet ab- 

 sorption disappears (3,4). Infrared spectrum given 

 in reference 4. [a]""' = —0.5° (c = 8.46 per cent 

 in water at pH 5.18). Rotation in 2 A' HCl changes 

 until a constant value of [a]^ = +9.7° is reached. 

 pKa' = 8.55. Positive ninhjdrin, sodium |3-naph- 

 thoquinone-4-sulfonate, and ammonium silver 

 nitrate tests (4). Crystallographic data given in 

 reference 4. Most stable at pH 6 to 8 (4). C5H7N3O4. 

 C = 34.85%; H = 4.30%; N = 24.44%; O = 37.36% 

 (4). Structural formula (5) is given in Chapter 6. 

 Azaserine has l^een synthesized, and both the DL- 

 and D -forms prepared (6). 



Biological activity: In vitro: Moderately active 

 on gram-positive and gram-negative bacteria, 

 mycobacteria, and fungi (2, 12). Amoebicidal to 

 Endamoeha histolytica at 50 to 100 pg per ml (22). 

 Active on Chlorella pyrenoidosa (27). Produces 

 greatly elongated, multinucleate, and nonseptate 

 filaments in E. coli at barely inhibitory levels and 

 higher (11). Induces formation of active phage 

 from lysogenic E. coli strain K-12 (17). The D- 

 form has no activity on Kloeckera brevis (6) or 

 other organisms (13). In vivo: A correlation be- 

 tween the activity of azaserine on K. brevis in 

 vitro and its activity on tumors in vivo was found 

 (2). Active on Plasmudiinn lophurae (chicks), 

 rickettsia of epidemic typhus (eggs), meningo- 

 pneumonitis virus (eggs), but not mycobacterial 

 or other viral infections (mice) (9). In mice: active 

 on 6C3HED lymphosarcoma (ascites and solid), 

 C3H-SX lymphoma (ascites and solid) (29); mod- 

 erately active on adenocarcinoma E0771, Patter- 

 son Ij-mphosarcoma, Mecca lymphosarcoma, sar- 

 coma ISO (ascitic and solid), Ehrlich ascites 

 carcinoma, and Krelis-2 ascites carcinoma; slightly 

 active on transmitted leukemia 82 and L1210 leu- 

 kemia (7, 19, 28). In rats: moderately active on 

 Walker carcinosarcoma 256, sarcoma R 39, Jen.^^en 

 sarcoma, and Murphy-Sturm lymphosarcoma; 

 slightly active on Flexner-Jobling carcinoma (8, 

 19, 28). Ascitic plasma cell neoplasm of mice 

 (70429) was initially inhibited l)ut later developed 



