190 



DESCRIPTIONS OF ANTIBIOTICS 



night. Impurities thus precipitated are discarded 

 and the acetone solution is concentrated in vacuo. 

 The concentrate is seeded and stored in the cold 

 (2), or freeze dried (1). Freeze -dried solid taken 

 up in hot ethanol, concentrated in vacuo, and 

 purified by chromatography on alumina (ben- 

 zene-ethanol, 1:1, as solvent and developer). 

 Crystallized from benzene-ethanol (1). Compo- 

 nent C is separated from A and B (see below) by 

 partition chromatography on cellulose (aqueous 

 n-butanol-1 per cent pyridine). A and B are sepa- 

 rated by countercurrent distribution (pyridine - 

 n-butanol-water, 1:100:100) (2). Repeated re- 

 crystallization from water (-1). 



Chemical and physical properties: Angustmycin 

 is a mixture of three substances, one of which 

 (A) is biologically active. The other two, B and 

 C, are biologically inactive. B was identified as 

 adenine, a structural moiety of A and C. Angust- 

 mycin A: Basic substance. Hydrate: m.p. 128- 

 130°C, remelting at 164.5-165.5°C (decomposition) 

 (4). Soluble in water, methanol, ethanol, pyri- 

 dine, acetic acid, methyl Cellosolve, dimethyl- 

 formamide, and phenol. Sparingly soluble in 

 butanol and dioxane. Insoluble in ether, acetone, 

 chloroform, carbon disulfide, ethyl acetate, and 

 other organic solvents (2). Positive Molisch test. 

 Negative Fehling and Tollen tests (4). Ultra- 

 violet absorption spectrum maximum at 260 ni/n 

 (e = 17,100) in acidic or alkaline solution or 

 methanol (1, 2, 4). Infrared spectrum given in 

 reference 4. [a]" = +17.02° (c = 1.4 per cent in 

 dimethylformamide) (2); pK,, = 9.8. CuHu04N5- 

 H.,0. C" = 44.4%; H = 5.09%; N = 23.42%. Struc- 

 tural formula of angustmycin A (6-amino-9 (L-1 , - 

 2-fucopyranoseenyl)purine) (4) given in Chapter 

 6. Tetraacetate: needles; m.p. 187-188°C (4). 

 Benzoate: m.p. 115-116°C (3). Hemimethanolate: 

 needles; m.p. 172-174°C. Acid hydrolysis products 

 include adenines and "angustose" or L-2-keto- 

 fucopyranose. Angustose crystallizes as needles; 

 m.p. 115-116°C. [at' = +18° (c = 1 per cent in 

 ethanol), CeHioOs - Rf (butanol-acetie acid- 

 water, 4:1:5) = 0.40 (3, 4, 6). Angustmycin C: 

 needles; m.p. 202-204°C (decomposition), [a]^^ = 

 — 71.1° (c = 1.8 per cent in pyridine). Ultraviolet 

 absorption spectrum maximum (water) at 260 

 van {Eil'm 510). Optically inactive. Infrared spec- 

 trum given in reference 5. Positive Molisch test. 

 Negative ninhydrin reaction (2, 5). CnHisOsNs . 

 Structural formula of angustmycin C (6-amino-9 

 (/3-p-psicofuranosyl)purine) (5) given in Chapter 

 6. An antibiotic, Antibiotic U 9586, said to have 

 the same structure as angustmycin C (inactive) 



was reported active on bacteria and tumors in 

 vivo (7). 



Chemical and physical properties of Antibiotic 

 U 9586: 6-Amino-9-D-psicofuranosylpurine. Nee- 

 dles; m.p. 212-214° (decomposition). Ultraviolet 

 absorption spectrum maxima at 259 m^i (E'lcm 

 508) in 0.01 A' H2SO4 and 261 mn (E^ 530) in 

 0.01 A^ NaOH. Positive ammoniacal silver nitrate 

 and Jordon-Prj'de tests (ketohexoses) and Bene- 

 dict test (after hydrolysis). Negative Bial, nin- 

 hydrin, and Benedict (before hydrolysis) tests. 

 [a]^ = —537° (c = 1 per cent in dimethyl sulf- 

 oxide). CuHisNsOs : C = 44.25%; H = 5.10%; 

 N = 23.74%; O = 27.02%. Acid hydrolysis prod- 

 ucts give adenine and D-psicose. Structural for- 

 mula given in Chapter 6. 



Biological activity: Angustmycin A inhibits 

 Mycobacterium 607 and M. phlei at 25 ng per ml. 

 Inactive against virulent M. tuberculosis H37Rv 

 at 100 ng per ml. No activity on other bacteria or 

 fungi (1). 



Toxicity: Mice tolerate 2.5 gm per kg intraperi- 

 toneally. 



References: 



1. Yimtsen, H. et al. J. Antibiotics (Japan) 



7A: 113-119, 1954. 



2. Yiintsen, H. et al. J. Antibiotics (Japan) 



9A: 195-201, 1956. 



3. Yiintsen, H. J. Antibiotics (Japan) llA: 



77-80, 1957. 



4. Yiintsen, H. J. Antibiotics (Japan) llA: 



233-243, 1957. 



5. Yiintsen, H. J. Antibiotics (Japan) llA: 



244-249, 1957. 



6. Yiint.sen, H. and Yonehara, H. Bull. Agr. 



Chem. Soc. Japan 21: 261-262, 1957. 



7. Schroeder, W. and Hocksema, H. J. Am. 



Chem. Soc. 81: 1767-1768, 1959. 



Anisomycin 



Produced by: Sireptoniyces griseolus (3, 4), S. 

 roseochromogenes (14), and a Streptonryces sp. 

 (3,4). 



Synonyms: Antibiotic PA 106; Flagecidin. 



Method of extraction: Broth-filtrate adjusted to 

 pH 9.0 and extracted countercurrently with 

 methyl isobutyl ketone. Back-extracted into 

 water at pH 2.0. This water is adjusted to pH 

 9.0 and extracted countercurrently with CCI4 . 

 On concentration of this extract, anisomycin 

 crystallizes out. Recrystallization from hot ethyl 

 acetate or water (2). Can also be extracted from 

 broth by diethyl ether, benzene, ethyl or butyl 

 acetate, butanol, or chloroform. May also be re- 

 covered and jjurified by adsorption on charcoal, 



