Baroud Milad, Lepeltier Elise, El-Makhour Yolla, Lautram Nolwenn, Bejaud Jerome, Thepot Sylvain, Duval Olivier
Micro & Nanomedecines Translationnelles (MINT), Inserm, The National Center for Scientific Research (CNRS), SFR ICAT, University of Angers, 49000 Angers, France.
Environmental Health Research Lab, Faculty of Science, Lebanese University, Nabatieh 1700, Lebanon.
Pharmaceuticals (Basel). 2021 Dec 17;14(12):1317. doi: 10.3390/ph14121317.
5-Azacitidine, a cytidine analogue used as a hypomethylating agent, is one of the main drugs for the treatment of myelodysplastic syndromes (MDSs) and acute myeloid leukemia (AML) in the elderly. However, after administration, it exhibits several limitations, including restricted diffusion and cellular internalization due to its hydrophilicity, and a rapid enzymatic degradation by adenosine deaminase. The aim of this study was to improve the drug cell diffusion and protect it from metabolic degradation via the synthesis of amphiphilic prodrugs and their potential self-assembly. Azacitidine was conjugated to two different omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The carboxylic acid group of the omega-3 fatty acids was effectively conjugated to the amine group of the azacitidine base, yielding two amphiphilic prodrugs. Nanoprecipitation of the obtained prodrugs was performed and self-assemblies were successfully obtained for both prodrugs, with a mean diameter of 190 nm, a polydispersity index below 0.2 and a positive zeta potential. The formation of self-assemblies was confirmed using pyrene as a fluorescent dye, and the critical aggregation concentrations were determined: 400 µM for AzaEPA and 688 µM for AzaDHA. Additionally, the stability of the obtained self-assemblies was studied and after 5 days their final stable arrangement was reached. Additionally, cryo-TEM revealed that the self-assemblies attain a multilamellar vesicle supramolecular structure. Moreover, the obtained self-assemblies presented promising cytotoxicity on a leukemia human cell line, having a low IC value, comparable to that of free azacitidine.
5-氮杂胞苷是一种用作低甲基化剂的胞苷类似物，是治疗老年骨髓增生异常综合征（MDS）和急性髓系白血病（AML）的主要药物之一。然而，给药后，它表现出几个局限性，包括由于其亲水性导致的扩散受限和细胞内化，以及被腺苷脱氨酶快速酶促降解。本研究的目的是通过合成两亲性前药及其潜在的自组装来改善药物的细胞扩散并保护其免受代谢降解。将氮杂胞苷与两种不同的ω-3脂肪酸，即二十碳五烯酸（EPA）和二十二碳六烯酸（DHA）偶联。ω-3脂肪酸的羧基有效地与氮杂胞苷碱基的胺基偶联，产生两种两亲性前药。对所得前药进行了纳米沉淀，两种前药均成功获得了自组装体，平均直径为190nm，多分散指数低于0.2，zeta电位为正。使用芘作为荧光染料确认了自组装体的形成，并测定了临界聚集浓度：AzaEPA为400μM，AzaDHA为688μM。此外，研究了所得自组装体的稳定性，5天后达到了最终的稳定排列。此外，冷冻透射电子显微镜显示自组装体具有多层囊泡超分子结构。此外，所得自组装体对白血病人类细胞系表现出有前景的细胞毒性，IC值较低，与游离氮杂胞苷相当。
