A novel microfluidic liposomal formulation for the delivery of the SN-38 camptothecin: characterization and in vitro assessment of its cytotoxic effect on two tumor cell lines.
A novel microfluidic liposomal formulation for the delivery of the SN-38 camptothecin: characterization and in vitro assessment of its cytotoxic effect on two tumor cell lines.
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DOI：
10.2147/ijn.s166219
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发表时间：
2018
影响因子：
8
通讯作者：
Mora M
中科院分区：
文献类型：
--
作者：
Casadó A;Sagristá ML;Mora M
伊立替康（CPT-11）和SN-38 -其活性代谢产物-是广泛用于各种癌症治疗方案的生物碱衍生的拓扑异构酶I相互作用化合物。为了解决与其内酯环在生理pH下的不稳定性和SN-38的极不溶性相关的问题，递送载体（例如脂质体）的开发已被认为是毫无疑问的医学兴趣的主题。这篇文章的重点是开发一种替代方案的经典脂质膜水化程序，以获得SN-38的药物制剂。通过微乳化法制备SN-38负载脂质体（SN-38 lip），无需预先制备脂质膜，并通过不同方法进行表征。通过光子相关光谱法确定处方参数，并通过吸收光谱法评价SN-38的包封率。通过冻干获得干燥白色粉末形式的SN-38 lip。进行MTT和LDH试验以评估脂质体（SN-38 lip）和溶解形式（SN-38 sol）的SN-38的细胞毒性作用;使用流式细胞术定量SN-38摄取并分析药物暴露后的细胞周期时相分布。制备了微流控、稳定、大小可控、带负电荷的脂质体SN-38，并将其掺入到蛋黄磷脂酰胆碱（EPC）/L-α-二油酰-磷脂酰丝氨酸（DOPS）（9：1）囊泡（SN-38 lip）中。最后得到SN-38 lip的冻干粉末，其易于复溶，同时保留理化参数。通过两种肿瘤细胞系（HeLa和Caco-2）的体外研究评估SN-38 lip的疗效，并与SN-38 sol进行比较。与SN-38 sol相比，它证明了SN-38 lip的最高摄取，与其最高细胞毒性效应一致。此外，脂质体制剂在两种细胞系中诱导了不同的细胞周期改变。结果突出了采购的SN-38脂质体制剂的潜在有用性，并为进行体内研究提供了基础，这些研究允许开发用于结直肠癌治疗的替代策略。
Irinotecan (CPT-11) and SN-38 – its active metabolite – are alkaloid-derived topoisomerase I interactive compounds widely used in various cancer therapy protocols. To solve the problems associated with the instability of their lactone ring at physiological pH and with the extreme insolubility of SN-38, the development of delivery carriers (eg, liposomes) has been considered a subject of unquestionable medical interest. This article focuses on the development of an alternative protocol to the classical lipid-film hydration procedures to obtain a pharmaceutical formulation for SN-38. SN-38-loaded liposomes (SN-38lip) were produced by microemulsification, without a prior lipid-film preparation step, and characterized by different methods. Formulation parameters were determined by photon correlation spectroscopy, and the SN-38 entrapment efficiency was evaluated by absorbance spectroscopy. SN-38lip was obtained as a dry, white powder by lyophilization. MTT and LDH assays were conducted to assess the cytotoxic effect of SN-38, both in liposomal (SN-38lip) and solubilized form (SN-38sol); flow cytometry was used to quantify SN-38 uptake and to analyze cell-cycle phase distribution after drug exposure. Microfluidic, stable, and controlled sized, negatively charged liposomes, with high SN-38 incorporation efficiency into egg yolk phosphatidylcholine (EPC)/L-α-dioleoyl-phospathidylserine (DOPS) (9:1) vesicles (SN-38lip), were prepared. A lyophilized powder of SN-38lip, easily reconstitutable while retaining physicochemical parameters, was finally obtained. The efficacy of SN-38lip was assessed by in vitro studies with two tumor cell lines (HeLa and Caco-2) and compared with that of SN-38sol. It demonstrated the highest uptake of SN-38lip, in accordance with its highest cytotoxicity effect, in comparison with that of SN-38sol. In addition, different cell-cycle alterations were induced in both cell lines by the liposomal formulation. The results highlight the potential usefulness of the procured SN-38 liposomal formulation and provide the basis for conducting in vivo studies that allow the development of alternative strategies for colorectal cancer treatment.