244 THE JOURNAL OF ANTIBIOTICS MAR. 1975

N-GLYCOSYL DERIVATIVES OF POLYENE butanol mixture, followed by washing with MACROLIDE ANTIBIOTICS acetone yielded 0.65 g of derivative exhibiting E 1% 1cm1350 at 382 nm. The minimum inhibi- Sir: tory concentration (MIC) against Saccharomvices The polyene macrolides are known as potent cerevisiae determined in liquid SABOURAUD antifungal agents. However their use in medium was 0.1 mcg/ml (MIC for parent am- therapy is limited by considerable toxicity photericin B, 0.08 mcg/ml). and very poor water solubility. Many efforts In the ultraviolet spectrum of the derivative had been taken to obvert these undesirable the positions of the absorption maxima and properties1-5). We report now a new group their relative intensities are identical with of derivatives of polyene macrolides exhibit- those of the parent antibiotic. The decrease ing improved solubility in water and organic of the E 1% 1cmvalues is in accordance with the solvents and retaining the biological activity increase of the molecular weight. of parent compounds. In the reaction of amphotericin B with These derivatives are prepared in the reac- glucose the substitution occurred on the amino tion of an antibiotic, containing a free group of mycosamine moiety. The derivative aliphatic amino group, with a carbohydrate treated with 1 % hydrogen chloride in meth- or its appropriate derivative. Optimal condi- ylene chloride afforded N-(1-deoxyfructose-1- tions of the reaction are the following: yl)-mycosamine. The structure of this com- dimethyl formamide as solvent, temperature pound follows from the mass spectral data of range 3540°C and 0.5 molar excess of carbo- its peracetylated dimethoxime. hydrate. The course of the reaction can be The N-glycosyl derivatives of polyene macro- followed by thin-layer chromatography on lides containing a free carboxyl group form silica gel with the solvent system, ethyl salts at pH values close to neutrality with acetate - acetic acid - water (4 : 1 : 1, v/v). The inorganic and organic bases. The salts are reaction is completed within 12~40 hours, readily soluble in water. The degree of their depending on the polyene macrolide and carbo- dispersion in aqueous solutions can be hydrate used. The products are isolated by estimated by comparing the electronic absorp- precipitation and washing with ethyl ether, tion spectra measured in water and in followed by drying under vacuum. The yields methanol6). are almost quantitative. The derivatives can The antifungal activities of N-glycosyl deriv- be purified by counter-current distribution atives are similar to those of the parent (chloroform - methanol - water, 2 : 2 : 1, v/v) polyene macrolides.7) Simultaneously they or butanol - ethyl acetate - methanol - water exhibit lower haemolytic activity, lower toxicity (20 : 10 : 5 : 35, v/v) or by partition chromato- and are effective in experimental candidiasis graphy on silica gel or Sephadex LH-20 in mice when administered intraperitoneally.7,8) (chloroform - methanol - water, 20 : 10 : 1, v/v). The encouraging biological properties of Derivatives of polyene macrolides represent- these compounds, along with their water solu- ing major structural groups (pimaricin, nys- bility indicate that N-glycosyl derivatives of tatin, amphotericin B, mycoheptin, candicidin, polyene macrolides are interesting novel anti- levorin and trichomycin) with various carbo- fungal chemotherapeutic agents. The sim- hydrates (such as glucose, mannose, fructose, plicity of their preparation is an additional ribose, maltose and glucuronic acid) were positive factor. synthesised and characterised. In an exampler synthesis, 1 g of amphotericin B (E 1% 1cm1480 Acknowledgements at 382 nm) and 0.3 g of glucose were dispersed We acknowledge the financial support of Polish in 15 ml of DMF and left for 16 hours at United Pharmaceutical Industry POLFA. The 37°C. The product was precipitated with authors would like, to express thanks to Phar- maceutical Factory Tarchomin-POLFA, Institute 300 ml of ethyl ether, centrifuged, washed 3 of Pharmaceutical Industry in Warsaw, Institute times with ether and dried in vacuum to ob- of Antibiotics in Leningrad, E.R. Squibb and tain 1.28g of crude derivative (E 1% 1cm1120 Sons, Inc., and Mycofarm Delft, for generous at 382 nm). Crystallization from methanol- gifts of polyene antibiotics. We are also indebted VOL. XXVIII NO. 3 THE JOURNAL OF ANTIBIOTICS 245 to Miss ELZBIETA BYLEC for technical assistance. ene macrolide derivatives. II. Physical- LEONARD FALKOWSKI, chemical properties of polyene macrolide JERZY GOLIK, esters and their water soluble salts. J. Anti- PAWEL KOLODZIEJCZYK, biotics 25 : 259260, 1972 JAN PAWLAK, 5) BRUZZESSE, T.; 1. BINDA, A. DI NARDO, G. JAN ZIELINSKI, GHIELMETTI & M. RIVA: Partricin methyl TADEUSZ ZIMINSKI, ester, a semisynthetic polyene antibiotic. EDWARD BOROWSKI Experientia 28 : 1515-.1516, 1972 6) FALKOWSKI, L.; A. MATULA & E. BOROWSKI: Department of Pharmaceutical Effect of polyene macrolide antibiotics on Technology and Biochemistry, Technical University, permeability of lyposomes. Advances in Antimicrobial and Antineoplastic Chemothe- Gdansk, Poland rapy, pp. 859860, Urban and Schwarzen- (Received April 24, 1974) berg, M(inchen, 1972 7) FALKOWSKI, L.; B. CYBULSKA, E. ZABLOCKA References & E. BOROWSKI: N-Glycosyl derivatives 1) BARTNER, E.; H. ZINNES, R. A. MOE & J. S. of polyene macrolide antifungal antibiotics. KULESZA: Studies on a new solubilised pre- I: in vitro studies. Advances in Antimi- paration of amphotericin B. Antibiot. Ann. crobial and Antineoplastic Chemotherapy 1957/1958 :53-58, 1958 (Proc. 8th Internat. Congr. Chemother., 2) LECHEVALIER, H.; E. BOROWSKI, J. 0. Athens, 1973), Urban and Schwarzenberg, LAMPEN& C. P. SCHAFFNER: Water soluble in press. N-acetyl derivatives of heptaene macrolide 8) FALKOWSKI, L.; H. BULUK, J. GOLIK, M. antifungal antibiotics: microbiological stud- BOBROWSKI, J. BOROWSKI & E. BOROWSKI; ies. Antibiot. & Chemoth. 11:640647, N-Glycosyl derivatives of polyene macrolide 1961 antifungal antibiotics. II. In vivo studies. 3) SCHAFFNER, C. P. & E. BOROWSKI: Biologi- Advances in Antimicrobial and Antineopla- cally active N-acyl derivatives of polyene stic Chemotherapy (Proc. 8th Internat. macrolide antifungal antibiotics. Antibiot. Congr. Chemother., Athens, 1973), Urban & Chemoth. 11 :724-732, 1961 and Schwarzenberg, in press. 4) SCHAFFNER, C. P. & W. MECHLINSKI: Poly-