DESCRIPTIONS OF ANTIBIOTICS 



249 



hexiinide, nystatin, and antitunioi' antibiotic 

 E73). 



Synonyms: Actidione (1), naramyoin A (71), 

 antibiotic A 67 (41). 



Hrniarks: Substances having great 1>' inferior or 

 no antifungal activity, inadonc (50, 51) and 

 nonactin (52), have been isolated from cyclohexi- 

 mide-containing broths. Streptovitacin, an anti- 

 tumor substance, has also been isolated from 

 certain cycloheximide broths (77). A cyclohexi- 

 midedike substance (m.p. 105-106°C) that was one 

 third as active a cycloheximide was reported (31). 

 It may be similar to naramycin B (see below). 



Method of extraction: I. Adsorbed on carl)on or 

 carbon-Celite at pH 2.0 or other acid pH from 

 broth-filtrate. Eluted with anhydrous or 80 per 

 cent acetone, or anhydrous methanol. Eluate dis- 

 tilled ofi', residual solution extracted with chloro- 

 form. Extract decolorized with carbon and evapo- 

 rated. Purified by countercurrent distribution 

 (benzene-water). Crystallized from amyl acetate 

 (1, 2), or from anhydrous ethyl ether on pouring 

 into petroleum ether. Further purified by chro- 

 matography on Darco (l-(iO-Celite, with 20 per 

 cent aqueous acetone as solvent and 60 to 100 per 

 cent acetone as developer. Crystallized from ac- 

 tive fractions on adding amyl acetate and seeding. 

 Recrystallized from hot water or 30 i^er cent 

 methanol (2). II. Broth-filtrate extracted with 

 amyl acetate. Mycelium extracted with acetone. 

 Extracts concentrated, then extracted with amyl 

 acetate. Amyl acetate-extracts combined and 

 concentrated in vacuo. Nonactin precipitates on 

 standing. Mother liciuor chromatographed on alu- 

 minum oxide and developed with chloroform- 

 met hanol (19:1). Further purified by countercur- 

 rent distribution (CCU-CHCL^-MeOH-water, 55: 

 55:75:25). Crystallized from ether by seeding; re- 

 crystallized from acetone-petroleum ether (52). 

 III. Adsorbed on carbon from broth-filtrates at 

 pH 5.5 to 6.0. P]luted with acetone at pH 4.0 to 

 4.5 followed by water. Eluates adjusted to pH 6 

 to 7, and acetone distilled off in vacuo. Aqueous 

 residue extracted into chloroform. Chloroform 

 neutralized and an oil which separates, removed. 

 Chloroform distilled in vacuo. Residue extracted 

 with water, clarified, and acetone added to 20 per 

 cent. Chromatography on Darco (j-60-Celite 545, 

 developed with 20 per cent acetone, and eluted 

 with aqueous acetone containing successively less 

 water. Active fractions are distilled in vacuo to 

 remove the acetone, extracted into chloroform, 

 and chloroform distilled off. Residue crystallized 

 from amy! acetate. Recrystallized from 30 per 

 cent methanol (53). 



Chemical and physical properties: Neutral sub- 

 stance. Rectangular or square colorless plates; 

 m.p. 115-117°C (1, 2, 53). Very soluble in all com- 

 mon organic solvents except saturated hydrocar- 

 bons. Soluble to 2.1 gm per 100 ml of water and 7 

 gm per 100 ml of amyl acetate (2). Ultraviolet 

 absorption spectrum maximum at 287 m/x (e = 

 36.7) (4). Infrared data given in reference 4. [aYo = 

 -2.8° (c = 9.6 per cent in methanol) (1), or +6.8° 

 (c = 2 per cent in water) (2). pKa' = 11-2 (4). 

 Positive Fearon-Mitchell test (primary or sec- 

 ondary alcoholic hydro.xyl) (4). Crystals stable 

 for several hours at 100°C. Acjueous solutions 

 stable for several hoiu's at 60 but not 70°C (26). 

 C = 64.167c-; H = 8.17%; N = 5.13%- C15H23NO4 . 

 Structural formula (2, 4) is given in Chapter 6. 

 /3 - [2 - (3 ,5 - dimethyl -2 - oxocyclohexyl)-2-hydroxy- 

 ethyl} glutarimide. Inactivated by alkali at room 

 temperature (1), giving a fragrant volatile prod- 

 uct, d-2,4-dimethylcyclohexanone, as well as pro- 

 pionaldehyde-2,2-diacetic acid (4). Cycloheximide 

 diacetate: m.p. 148-149°C (2). [a]f = +24.6° (c = 

 3.7 per cent in methanol). Biologically inactive 

 (1). Monoxime: m.p. 203-204°C (2, 4). Semicarba- 

 zone: m.p. 182-183 °C (2, 4). 



Biological activity: In vitro: Active on a variety 

 of plant pathogens at 0.125 to 20 Mg per ml, includ- 

 ing members of the phycomycetes, ascomycetes, 

 basidiomycetes, and Fungi Imperfecti (8). Most 

 active on yeasts of the Saccharomyces genus; less 

 so on Torula or Hansensia. Not active on Kloeckera 

 apiculata. Active on most alcoholic fermentation 

 processes; slightly active on respiration; not ac- 

 tive on lactic fermentation (31). Not active on 

 gram-positive or gram-negative bacteria (15). 

 Active on Tetrahymena geleii, Euglena gracilis var. 

 hacillaris (colorless), and Endamoeba histolytica 

 (25). Inhibits the growth of certain members of 

 Chlorophyceae, Xanthophyceae, and Bacillario- 

 phyceae at 50 ppm or less, but has no effect on 

 Myxophyceae (68). Active in vitro on influenza A 

 (PR 8) virus liy direct inactivation of the virus 

 particles (67). Partially inhibitory levels of cyclo- 

 heximide in a medium on which Allomyces arbus- 

 cula is grown give rise to the formation of male 

 and female gametangia on portions of the sporo- 

 phyte (22). Inhibits protein and nucleic acid syn- 

 thesis in Sacch. carlsbergensis. At minimal growth- 

 inhibitory levels, synthesis of desoxyribonucleic 

 acid and protein is completely inhibited, but ribo- 

 luicleic acid synthesis continues slowly for some 

 time ((id). In animals: Temporary clearance of E. 

 histolytica from macaques (13), rats, and dogs 

 (58). Inhibits growth of the following in mice, 

 rats, or hamsters: Miyono adenocarcinoma, Mecca 



