Onishi Reina, Watanabe Ayahisa, Nakajima Mado, Sekiguchi Mitsuaki, Kugimiya Akira, Kinouchi Hiroki, Nihashi Yoichiro, Kamimori Hiroshi
Pharmaceutical Research Division, Shionogi & Co., Ltd.
Anal Sci. 2015;31(12):1255-60. doi: 10.2116/analsci.31.1255.
In the present study, we developed an assay to evaluate the kinetic binding properties of the unconjugated antisense oligonucleotide (ASO) and lipophilic and hydrophilic ligands conjugated ASOs to mouse and human serum albumin, and lipoproteins using surface plasmon resonance (SPR). The lipophilic ligands conjugated ASOs showed clear affinity to the albumins and lipoproteins, while the unconjugated and hydrophilic ligand conjugated ASOs showed no interaction. The SPR method showed reproducible immobilization of albumins and lipoproteins as ligands on the sensor chip, and reproducible affinity kinetic parameters of interaction of ASOs conjugated with the ligands could be obtained. The kinetic binding data of these ASOs to albumin and lipoproteins by SPR were related with the distributions in the whole liver in mice after administration of these conjugated ASOs. The results demonstrated that our SPR method could be a valuable tool for predicting the mechanism of the properties of delivery of conjugated ASOs to the organs.
在本研究中，我们开发了一种分析方法，以使用表面等离子体共振（SPR）评估未缀合的反义寡核苷酸（ASO）以及与亲脂性和亲水性配体缀合的ASO与小鼠和人血清白蛋白及脂蛋白的动力学结合特性。与亲脂性配体缀合的ASO对白蛋白和脂蛋白表现出明显的亲和力，而未缀合的以及与亲水性配体缀合的ASO则没有相互作用。SPR方法显示白蛋白和脂蛋白作为配体可重复固定在传感器芯片上，并且可以获得与配体缀合的ASO相互作用的可重复的亲和动力学参数。通过SPR得到的这些ASO与白蛋白和脂蛋白的动力学结合数据与这些缀合的ASO给药后在小鼠全肝中的分布有关。结果表明，我们的SPR方法可能是预测缀合的ASO向器官递送特性机制的有价值工具。
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