DESCRIPTIONS OF ANTIBIOTICS 



22?. 



ness in vacuo. Residue dissolved in methyl alcohol; 

 decolorization with charcoal. Filtrate evaporated 

 to dryness, dissolved in anhydrous ethyl acetate, 

 and passed through a column of alumina. Column 

 washed with ethyl acetate, acetone, and methanol. 

 Elution with phosphate buffer at pH 6. Adjusted 

 to pH 3, extracted with ethyl acetate, washed with 

 water, and concentrated in vacuo. A precipitate 

 forms, which is dissolved in ether; petroleum ether 

 added gradually. Upon standing at room tempera- 

 ture, colorless crystals are obtained. Recrystalli- 

 z at ion from the same ether-petroleum ether mix- 

 ture. 



Chemical and phijsical properties: Colorless crys- 

 tals; m.p. 66-67°C. Soluble in water, alcohols, ace- 

 tone, and ether. Moderateh^ soluble in benzene. 

 Insoluble in petroleum ether. C = 30.8-4%; H = 

 4.40%; N = 11.66%,. C.3H5O4N. Molecular weight 

 119.8 (foimd, Rast's method, 110). Light-absorp- 

 tion maximum at about 270 iw/x at pH 8.4. Infrared 

 absorption spectrum given in reference 1. Bovino- 

 cidin was found to be identical to |8-nitropropionic 

 acid. 



Biological activity: Weakly active against -1/. 

 tuberculosis BCG. No activity against other myco- 

 bacteria, bacteria, and fungi tested. Biological 

 activity not reduced by /3-alanine. 



Toxicity: LD50 (mice) 50 mg per kg intrave- 

 nously. 



Reference: 1. Anzai, K. and Suzuki, S. J. Anti- 

 biotics (Japan) 13A: 133-136, 1960. 



Broni tetracycline 



Produced by: Certain Streptomyces aureofaciens 

 (chlortetracycline-producer) mutants. Produced 

 instead of chlortetracycline when bromide is pres- 

 ent in a Cl-deficient medium (3). 



Method of extraction: Broth-filtrate acidified to 

 pH 2.0, mixed with Hyflo Super-Cel, and filtered. 

 BaCl-.-2H20 added to filtrate, pH adjusted to 8.5, 

 and cooled to 0°C. Solids extracted with butanol 

 at pH 2.0. Extract adjusted to pH 3.5 and concen- 

 trated in vacuo at <30°C. Concentrate adjusted 

 to pH 5.0; precipitate washed with ether and dried 

 (1). Separated from concurrently produced tetra- 

 cj^'cline by partition chromatography (butanol- 

 chloroform over stationary Celite-aqueous HCl) 

 (3). Purification by precipitation as HCl salt and 

 conversion to base by Vjringing an aqueous solu- 

 tion to pH 5.2 with triethylamine and cooling. 

 Crystallized from benzene (1). 



Chemical and physical properties: Base: m.p. 

 170-1 72°C. [cx]l° = -196° (in 0.1 X HCl). Ultra- 

 violet absorption spectrum maxima (in 0.1 A HCl) 

 at 227 niM (e = 17915), 260 m^ (t = 17480), and 



370 m/x (t = 9630) ; or (in 0.25 -Y NaOH) at 225 

 niM (e = 28215), 255 m^ (e = 15115), 285 m^ (e = 

 14610), and 345 mn (« = 7221). Infrared spectrum 

 is the same as that of chlortetracycline (1). More 

 stable than the other tetracyclines in distilled 

 water at 100°C for 15 minutes. CasHosOsNoBr (1, 

 2). Structural formula given in Chapter 6. HCl 

 salt: Bright ^-ellow crystals. Browns at 218°C; 

 decomposes at 235°C. Soluble in water to 1.36 per 

 cent at 25°C, in dry butanol to 0.038 per cent. 

 I)Ka = 3.4, 7.4, and 9.2. [a]„ = -205° (0.5 per cent 

 in 0.03 X aqueous HCl). Yields tetracycline on 

 catalytic hydrogenation. Has 77 per cent of the 

 absorption of chlortetracycline at 368 m^ (2, 3). 



Biological activity: Qualitatively, antibacterial 

 activity resembles chlortetracycline. Has 95 per 

 cent of the activity of chlortetracycline on Staph, 

 aureus on a weight basis, and 90 per cent of the 

 activity on E. coli (3). Has same activity in vivo 

 as other tetracyclines (2). 



Toxicity: LD50 (HCl salt, mice) 89 mg per kg 

 intravenously (2). 



References: 



1. Sensi, P. ct al. Farmaco (Pavia) 10: 337- 



345, 1955. 



2. Rolland, G. et al . Farmaco (Pavia) 1(1: 



340-355, 1955. 



3. Doerschuk, A. P. et al. J. Am. Chem. Soc. 



78: 1508-1509, 1956. 



B 



rvanivcin 



Produced by: Streptomyces hawaiiensis (1, 2). 



Synonym: Thiactin (2). 



Method of extraction: Extracted from whole 

 broth or V)roth-filtrate with methyl isobutyl ke- 

 tone, butanol, or chloroform. Purification b}- ad- 

 sorption on alumina or Florisil from chloroform; 

 washing with water, acetone, and chloroform; 

 and elution with a 5 per cent methanol-95 per cent 

 chloroform mixture or aciueous t-butyl alcohol. 

 Original methyl isobutyl ketone extract (in the 

 cold) can be processed ))y "flashing" into water 

 (cUstilling (71 vacuo with the addition of water 

 until no original solvent remains) and lyophiliz- 

 ing, or In- flashing into t-butyl alcohol. Recrys- 

 tallized by flashing a concentrated chloroform 

 solution into methyl isobutyl ketone, or by cooling 

 a warm methyl Cellosolve or dio.xane solution and 

 addition of 10 to 50 per cent water or acetone. 

 Forms salts with mineral acids or metals such as 

 calcium or sodium (1, 2). 



Chemical and physical properties: Amphoteric, 

 white, crystalline, sulfur-containing polypeptide. 

 Darkens at about 205-224°C and melts at 223- 

 235°C. Soluble to the extent of >2 mg per ml in 



