Analysis of the interaction of cyclosporine congeners with cell membrane models
Analysis of the interaction of cyclosporine congeners with cell membrane models
复制标题
环孢素同系物与细胞膜模型相互作用的分析
DOI：
10.1016/j.jpba.2022.114874
复制
发表时间：
2022
影响因子：
3.4
通讯作者：
Sakai-Kato Kumiko
中科院分区：
文献类型：
--
作者：
Nakao Mizuka;Takechi-Haraya Yuki;Ohgita Takashi;Saito Hiroyuki;Demizu Yosuke;Izutsu Ken-Ichi;Sakai-Kato Kumiko
由于大环肽的相对高分子量，研究细胞摄取机制是有效设计大环肽作为潜在药物所必需的。我们以前曾报道，使用HPLC，环孢素A，模型大环肽，和它的同系物B，C，和D有不同的亲脂性，尽管只有一个氨基酸不同。在本研究中，我们研究了亲脂性的这种差异如何影响同源物与细胞膜的相互作用。圆二色光谱表明，即使在高温下，四种同系物的二级结构也是相似的。作为细胞膜模型，脂质体存在下的四种同系物的摩尔椭圆率不同，环孢素D和A表现出较低的摩尔椭圆率，而环孢素C表现出较高的摩尔椭圆率。使用Laurdan的荧光光谱分析表明，脂质体水合作用在环孢菌素，尤其是环孢菌素D和A的存在下降低。HPLC定量分析也表明，环孢素D在HpG2细胞中的内化量最大。我们确定，使用光谱和HPLC，同源物与细胞膜的相互作用的强度总体上与来自每个同源物的侧链的亲脂性相关。我们的研究结果将有助于设计新的大环肽具有良好的药物性能。
Owing to the relatively high molecular weight of macrocyclic peptides, investigation of the cellular uptake mechanism is required for the efficient design of macrocyclic peptides as potential drugs. We have previously reported, using HPLC, that cyclosporine A, a model macrocyclic peptide, and its congeners B, C, and D had different lipophilicity despite differing by only one amino acid. In the present study, we investigated how this difference in lipophilicity affected the interaction of the congeners with cell membranes. The circular dichroism spectra showed that the secondary structures were similar between the four congeners even at high temperature. The molar ellipticity of the four congeners in the presence of liposomes, as a cell membrane model, differed, and cyclosporines D and A showed lower molar ellipticity, while cyclosporine C exhibited higher molar ellipticity. Fluorescent spectra analysis using Laurdan indicated that liposome hydration was decreased in the presence of the cyclosporines, especially cyclosporines D and A. HPLC analysis also quantitatively showed that the amount of cyclosporine molecules internalized in HpG2 cells was the largest for cyclosporine D. We determined, using spectroscopy and HPLC, that the intensity of the interaction of the congeners with cell membranes was overall correlated with the lipophilicity derived from the side chains of each congener. Our results will contribute to the design of new macrocyclic peptides with favorable drug properties.