Amphotericin B (AmB) is a very potent antibiotic which still remains as the gold standart for the treatment of systemic fungal infections. AmB is a member of Biopharmaceutical Classification System Class IV, mainly characterized by its poor solubility and low permeability. In this study, AmB/AmB-α cyclodextrin complex double loaded liposomes were developed using the design of experiments (DoE®) approach to optimize/determine the effects of lipid composition and other parameters on final product properties such as encapsulation efficacy, particle size, polidispersity index and zeta potential. Experimental design 24 was used for optimization of these properties in which four factors were studied in two levels. Double loaded liposomes showed much higher physical stability than liposomes loaded only with free AmB by the means of particle size, zeta potential and encapsulation efficiency, in addition exhibited sustained release of AmB over 72 h (26.7%) with faster onset time. On the other hand, 4-fold improved antimicrobial efficiency, Minimum Inhibitory Concentration (0.125 µg/mL) and Minimum Fungucidal Concentration (0.5 µg/mL) was determined by double loaded liposomes against C.albicans compared to Ambisome®. Dose dependent effects of the double loaded liposomes were investigated by cytotoxicity studies on Vero and L-929 cells. No significant cytotoxicity observed for AmB/AmB-αCD complex double loaded liposomes and Ambisome at tested concentrations while free AmB caused severe cytotoxicity. Lastly the developed double loaded liposomes did not cause an increase in NGAL (an early biomarker for akute kidney toxicity) levels for both Vero and HK-2 cell lines compared to free AmB.

中文翻译：

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使用析因设计开发有效的 AmB/AmB-αCD 复合双负载脂质体用于全身性真菌感染治疗

两性霉素 B (AmB) 是一种非常有效的抗生素，仍然是治疗全身性真菌感染的黄金标准。AmB是生物制药分类系统IV类的成员，主要特点是溶解性差，渗透性低。在本研究中，使用实验设计 (DoE®) 方法开发了 AmB/AmB-α 环糊精复合物双载脂质体，以优化/确定脂质成分和其他参数对最终产品特性的影响，例如封装功效、粒径、多分散指数和 zeta 电位。实验设计 24 用于优化这些特性，其中在两个水平上研究了四个因素。双负载脂质体比仅负载游离 AmB 的脂质体显示出更高的物理稳定性，通过粒径，zeta 电位和包封效率，此外还表现出 AmB 的持续释放超过 72 小时（26.7%），并且起效时间更快。另一方面，与 Ambisome® 相比，双负载脂质体针对白色念珠菌测定的抗菌效率、最低抑菌浓度 (0.125 µg/mL) 和最低杀菌浓度 (0.5 µg/mL) 提高了 4 倍。通过对 Vero 和 L-929 细胞的细胞毒性研究来研究双负载脂质体的剂量依赖性效应。在测试浓度下，AmB/AmB-αCD 复合物双载脂质体和 Ambisome 未观察到显着的细胞毒性，而游离 AmB 引起严重的细胞毒性。最后，与游离 AmB 相比，开发的双负载脂质体不会导致 Vero 和 HK-2 细胞系的 NGAL（急性肾毒性的早期生物标志物）水平增加。