216 



DESCRIPTIONS OF ANTIBIOTICS 



8. Maeda, K. J. Antibiotics (Japan) 6A: 



137-138, 1953. 



9. Celmer, W. D. and Solomons, LA. Anti- 



biotics Ann. 022-625, 1953-1954. 



10. Takahashi, T. et al. J. Antibiotics (Japan) 



7A: 26, 1954. 



11. Celmer, W. D. and Solomons, I. A. J. Am. 



Chem. Soc. 77: 2861-2865, 1955. 



12. Nitta, K. et al. J. Antibiotics (Japan) 



8A: 120-125, 1955. 



13. Koaze, Y. et al. J. Antibiotics (Japan) 



9A: 89-96, 1956. 



14. Umezawa, H. Giorn. microbiol. 2:160- 



193, 1956. 



15. Nakamura, M. et al. Ann. Rept. Takamine 



Lab. 9:35-43,1957. 



16. Funaki, M. et al. J. Antibiotics (Japan) 



llA: 143-149, 1958. 



Ayfactin 



Produced by: Strains of Streptomyces aureofa- 

 ciens and S. viridifaciens which produce tetra- 

 cycline and/or chlortetracycline. 



Synonyms: Antibiotic AYF and aureofacin are 

 probably identical with ayfactin. 



Method of extraction: I. Whole broth acidified 

 to pH 2.0 and filtered with a filter-aid. Mycelium 

 adjusted to pH 9.0 to 10.0 with NH4OH and ex- 

 tracted with n-butanol. Extract washed with 

 water, adjusted to pH 7.0, and concentrated by 

 azeotropic distillation, preferably in vacuo. Ayfac- 

 tin precipitates on cooling or on addition of Skelly- 

 solve C. Purified by: (a) Slurrying with methyl 

 isobutyl ketone and water. Precipitate taken up 

 in pyridine, solution concentrated to dryness in 

 vacuo, and toluene added to precipitate ayfactin 

 from the residue. Precipitate dissolved in di- 

 methylformamide and filtered to remove insoluble 

 impurities. Precipitated from filtrate with Skelly- 

 solves, isopropanol, or toluene, (b) Slurrying with 

 aqueous acid, hot formamide, methanol, or ace- 

 tone. II. Mycelium obtained as in I; extracted 

 with acetone. Extract adjusted to pH 6.0 to pre- 

 cipitate ayfactin. III. Purified by suspending 

 crude substance in a water-n-butanol (1:1) mix- 

 ture, adjusting to pH 1.5 with stirring, and collect- 

 ing the active precipitate formed at the interface. 

 Butanol-phase extracted with water at pH 7.0. 

 Aqueous extract lyophilized to give ayfactin. 

 Butanol concentrated, and ayfactin precipitated 

 on addition of Skelh'solve C. 



Chemical and physical properties: Crystalline 

 heptaene. Does not sublime in vacuo up to 200°C. 

 Nearly insoluble in water. Very soluble in n-buta- 

 nol, dimethylformamide, pj^ridine, morpholine, 



and piperidine. Insoluble in ethyl acetate, ace- 

 tone, ether, methanol, chloroform, and HCl. 

 Ultraviolet absorption spectrum maxima (pyri- 

 dine) at 330, 348, 366, 388, and 410 ni/u- Infrared 

 spectrum given in reference 1. C = 64.85%; H = 

 7.68%; N = 3.01%. C25H36-36NO7 . 



Biological activity: Probably active on fila- 

 mentous fungi and yeasts. Little antibacterial 

 activity. Active in protecting mice from C. albi- 

 cans infections. Not absorbed from the gastro- 

 intestinal tract. 



Toxicity: LDsu (mice) 0.8 mg per kg intraperi- 

 toneally. 



Reference: 1. British Patent 796,982, June 25, 

 1958. 



Azalomycins 



Produced by: Streptomyces hygroscopicus K5-4, 

 which also produces an antimycobacterial factor. 



Synonyms: Azalomycin F has properties in 

 common with musariu and hygrostatin. Azaolo- 

 mycin B is similar to elaiophylin. 



Method of extraction: I. Extraction of the wet 

 mycelial cake with acetone. Evaporation of ace- 

 tone in vacuo. Residue dissolved in methanol, 

 which is poured into 5 times its volume of ether. 

 The insoluble part is mainly azalomycin F; the 

 fraction soluble in the methanol-ether mixture is 

 mainly azalomycin B and the antimycobacterial 

 factor. II. Extraction of the l>roth-filtrate with 

 ethyl acetate yields azalomycin B; extraction 

 with butanol yields azalomycin F (1). Azalomycin 

 B, containing solvent solution from either the 

 mycelium or the broth, is concentrated to dryness 

 in vacuo, dissolved in ethyl acetate, washed with 

 2 per cent NaHCOs , 0.01 A' HCl, and water, and 

 concentrated in vacuo. Upon standing overnight 

 at 5-10°C, crystals of azalomycin B precipitate. 

 Recrystallization from aqueous alcohol or acetone 

 (1). Azalomycin F: Crude extract of azalomycin F 

 is dissolved in methanol and chromatographed 

 over a column of alumina. Elution with methanol 

 or 20 per cent aqueous methanol. Eluate is con- 

 centrated. Upon addition of acetone, the anti- 

 biotic precipitates. Recrystallization from meth- 

 anol -acetone solutions and finally from methanol 



(1). 



Chemical and physical properties: Azalomycin 

 B: Neutral compound. White, needle-shaped 

 crystals; m.p. 185-187°C (decomposition). C = 

 61.88%o; H = 8.72%,; OCH, = 10.12%,. Molecular 

 weight (Rast) 284. C,4H2405 • Molecular weight 

 272.33. [aif = -48° (c = 1 per cent in methanol). 

 Light-absorption maximum 252.5 ni/i in methanol. 

 Infrared absorption spectrum given in reference 



