228 



DESCRIPTIONS OF ANTIBIOTICS 



Biological activity: Very active on fungi, yeasts, 

 and Trichomonas vaginalis (1). Active in rats on 

 Yoshida sarcoma cells, but does not prolong sur- 

 vival time. Not active on Ehrlich carcinoma 

 (mice) (2). 



Toxicity: LD50 5 mg per kg sul)cutaneously ; 

 animal not given. 



References: 



1. Shibata, M. et al. J. Antil)iotics (Japan) 



7B: 168, 1954. 



2. Aramaki, Y. et al. Ann. Rept. Takeda Re- 



search Lab. 14: 60-91, 1955. 



Carboniyciii 



Produced by: Sireptomyces halstedii (1, 26) (also 

 produces carbomycin B), S. hygroscopicus (8), and 

 S. albireticuli (21). 



Synonyms: Magnamycin; carljomycin A, anti- 

 biotic M4209 (9). 



Methods of extraction: I. Broth-filtrate ex- 

 tracted with benzene, chloroform, amyl acetate, 

 trichloroethylene, butanol, or ether at pH 6.6 or 

 higher. Solvent: (A) Evaporated to dryness. Resi- 

 due yields crystals when taken up in a small 

 amount of isopropanol. Recrystallized from aque- 

 ous acetone or ethanol. (B) Solvent cHstilled off 

 in the presence of water. Watery residual solution: 

 (a) freeze dried and recrystallized from benzene- 

 petroleum ether or aqueous methanol; (b) chro- 

 matographed on silica gel from 10 per cent acetone 

 in benzene and developed with acetone-benzene 

 mixtures containing gradually increasing amounts 

 of acetone, the principal fraction being in the 30 

 per cent acetone in benzene; solvent evaporated 

 off; residue crystallized from 70 per cent hot aque- 

 ous isopropanol on addition of water; or (c) ex- 

 tracted with neutral benzene, extract concen- 

 trated; concentrate back-extracted into water at 

 pH 2.0 to 2.5; aqueous extract neutralized to give 

 free base (8, 9, 22). II. Broth-filtrate extracted 

 with methyl isobutyl ketone at pH 3.0. Extract 

 concentrated in vacuo and extracted with dilute 

 sulfuric acid (pH 2.0). Aqueous extract washed 

 with benzene, adjusted to pH 6.5, and extracted 

 with ether repeatedly. Ether evaporated to dry- 

 ness. Residue triturated in ethanol. Recrystallized 

 from methanol-water (10). III. Carbomycin 

 forms complexes with such aromatic solvents as 

 benzene, mesitylene, benzyl chloride, toluene, 

 ethyl benzene, naphthalene, chlorobenzene, and 

 toluene. Such complexes may be precipitated by 

 addition of hexane to a sufficiently pure solution of 

 the complex or V)y concentration in vacuo. The 

 complex may be cleaved to give the free base as 

 shown previously (24). These comple.xes may also 



be separated as solids from the crude broth-ttltrate 

 if sufhcient complexing agent is added at pH 8.5 

 to 9.5. Cleavage is obtained with acid, or the com- 

 plex extracted into acetone or methanol and the 

 extract concentrated to remove the solvent. Base 

 precipitates on addition of water to the residue 

 (25). 



Chemical and physical properties: Weakly basic 

 (26). Macrolide (28). Base: Various crystal forms 

 have been reported. Prisms; m.p. 207-209°C (open 

 capillary in oil bath) or 220-222°C (hot stage) 

 (22); slender white blunt-ended needles; m.p. 

 199. 5-200. 5°C (1); or rectangular plates or laths; 

 softens at 208-210°C; m.p. 210-216°C (decomposi- 

 tion) (9, 10). Very soluble in chloroform, ethyl 

 acetate, glacial acetic acid, 0.05 N HCl, amyl ace- 

 tate, Initanol, benzene, and acetone. Less soluble 

 in methanol, ethanol (25 mg per ml), ethyl ether, 

 and isopropanol (6.2 mg per ml). Insoluble in hot 

 or cold water, aqueous alkali, and petrolevmi ether 



9, 22). Ultraviolet spectriuii maxima at 238 nifi 

 (£■!";, 185) and 327 m/x (E\ln 0.9) in absolute etha- 

 nol (10) or at 241 m/x (£'1L 158) in 2 per cent Na2- 

 HP()4 (22). Infrared spectrum given in references 



10, 22, and 26. [a]t = -53° ± 3° (c = 0.3 per cent 

 in 0.05 N acetic acid), or —56.1° (c = 1 per cent in 

 chloroform) (10, 22). Positive 2,4-dinitrophenyl- 

 hydrazine, Bayer permanganate, bromine, and 

 eerie nitrate tests. Positive (10) or negative (22) 

 Fehling and Tollen tests. Violet color with Schiff's 

 fuchsin reagent (9). Violet color in 4 A^ HCl (10) 

 or red-violet with strong acid (5). Intense yellow 

 color in methanolic KOH (9). Negative ninhydrin, 

 Van Slyke, boric acid, and FeCls tests (10). Water- 

 insoluble precipitate with trichloroacetic acid (9). 

 No precipitate with silver nitrate or mercuric 

 chloride (22). Rf values in various systems given in 

 references 8 and 22. Most stable in aqueous solu- 

 tions at pH 5 to 7; less stable at pH 3 or 9. Crystals 

 stable for several months in the dark at room tem- 

 perature (10). C = 59.89%; H = 7.96%; N = 1.78%; 

 C— CHs = 10.58%; N— (CH3)2 = 2.94%; OCH., 

 = 3.75% (12). No S, P, or halogen (5). Molecular 

 weight 810 to 866 (9, 12, 26). C^HejOieN. Struc- 

 tural formula is given in Chapter 6. Mild acid 

 hydrolysis yields, among other products, the 4- 

 isovaleryl methyl glycoside of the sugar mycarose. 

 The glycoside is an oily neutral substance; b.p 

 116°C (1.1 mm), nf 1.4493; [«]'„" = -10.7° (c = 

 9 per cent in chloroform). Mycarose itself is a 

 crystalline solid; m.p. 128-129°C. [a]f = -31.1° 

 (c = 4 per cent in water) (11). Strong acid hydroly- 

 sis yields miicaminose, CSH17NO4 . Hydrochlo- 

 ride: m.p. 115-116°C. [aif = -f-31° (c = 1 per cent 

 in water) (20). Methanolysis removes mycarose, 



