Department of Biological Science, Graduate School of Science, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan.
Interprotein Corporation, 3-10-2 Toyosaki, Kita-ku, Osaka 531-0072, Japan.
Mol Pharm. 2022 Jul 4;19(7):2279-2286. doi: 10.1021/acs.molpharmaceut.2c00106. Epub 2022 May 28.
The effectiveness of protein and peptide pharmaceuticals depends essentially on their intrinsic pharmacokinetics. Small-sized pharmaceuticals in particular often suffer from short serum half-lives due to rapid renal clearance. To improve the pharmacokinetics by association with serum albumin (SA) , we generated an SA-binding tag of a helix-loop-helix (HLH) peptide to be linked with protein pharmaceuticals. For use in future preclinical studies, screening of yeast-displayed HLH peptide libraries against human SA (HSA) and mouse SA (MSA) was alternately repeated to give the SA-binding peptide AY-VE, which exhibited cross-binding activities to HSA and MSA with of 65 and 20 nM, respectively. As a proof of concept, we site-specifically conjugated peptide AY-VE with insulin to examine its bioactivity . In mouse bioassay monitoring the blood glucose level, the AY-VE conjugate was found to have a prolonged hypoglycemic effect for 12 h. The HLH peptide tag is a general platform for extending the bioactivity of therapeutic peptides or proteins.
蛋白质和肽类药物的疗效在很大程度上取决于其内在的药代动力学特性。由于肾脏清除速度快，特别是小尺寸的药物往往血清半衰期较短。为了通过与血清白蛋白（SA）结合来改善药代动力学性质，我们生成了一个螺旋-环-螺旋（HLH）肽的 SA 结合标签，用于与蛋白质药物偶联。为了用于未来的临床前研究，我们交替筛选酵母展示的 HLH 肽文库与人 SA（HSA）和鼠 SA（MSA）的结合，得到了 SA 结合肽 AY-VE，它对 HSA 和 MSA 的结合活性分别为 65 和 20 nM。作为概念验证，我们将肽 AY-VE 定点偶联到胰岛素上，以研究其生物活性。在监测血糖水平的小鼠生物测定中，发现 AY-VE 缀合物具有长达 12 小时的延长降血糖作用。HLH 肽标签是延长治疗性肽或蛋白质生物活性的通用平台。
