Mild heating of amphotericin B-desoxycholate: Effects on ultrastructure, in vitro activity and toxicity, and therapeutic efficacy in severe candidiasis in leukopenic mice

Heated (20 min at 70 degrees C) amphotericin B-desoxycholate (hAMB-DOC) was further characterized, as was another formulation obtained after centrifug ation (60 min, 3000 x g), hcAMB-DOC. Conventional AR-IB-DOC consisted of in dividual micelles (approximately 4 nm in diameter) and threadlike aggregate d micelles, as revealed by cryo-transmission electron microscopy, For both hAMB-DOC and hcAMB-DOC, pleiomorphic cobweb structures were observed with a mean particle size of approximately 300 nm as determined by laser diffract ion. The potent antifungal activity of AMB-DOC against Candida albicans is not reduced by heating. Effective killing of C. albicans (>99.9% within 6 h ) was obtained at 0.1 mg/liter with each of the AMB formulations. For AMB-D OC, hAMB-DOC, and hcAMB-DOC, cation release (Rb-86(+)) from C. albicans of greater than or equal to 50% was observed at 0.8, 0.4, and 0.4 mg/liter, re spectively. After heating of AMB-DOC, toxicity was reduced 16-fold as deter mined by red blood cell (RBC) lysis, For AMB-DOC, hAMB-DOC, and hcAMB-DOC, hemolysis of greater than or equal to 50% was observed at 6.4, 102.4, and 1 02.4 mg/liter, respectively. In contrast, AMB-DOC and its derivates showed similar toxicities in terms of cation release from RBC, Ear AMB-DOC, hAMB-D OC, and hcAMB-DOC, cation release (Rb-86(+)) of greater than or equal to 50 % was observed at 1.6, 0.8, and 0.8 mg/liter, respectively. In persistently leukopenic mice with severe invasive candidiasis, higher dosages of both h AMB-DOC and hcAMB-DOC were tolerated than those of conventional AMB-DOC (3 versus 0.8 mg/kg of body weight, respectively), resulting in significantly improved therapeutic efficacy, In conclusion, this new approach of heating AMB-DOC may be of great value for further optimizing the treatment of sever e fungal infections.