基于丙肝病毒NS3/4A蛋白酶及其靶向药物Danoprevir的小分子介导的异源二聚化的蛋白质元件的设计与合成

合成生物学已被应用于新型的临床治疗模式，如CAR-T细胞疗法已能够成功治疗白血病和淋巴瘤。但是现有的CAR-T细胞一旦进入病人体内，因其功能无法被精准调控，其过度激活可能造成严重危害甚至致命。使用小分子药物作为控制细胞功能的开关，在时间和空间上精准地调控CAR-T细胞的功能的方法，有望解决CAR-T细胞疗法的安全性问题。目前已有一些研究成功运用小分子化合物介导的二聚化生物元件来控制CAR-T细胞的开关。然而这些小分子化合物均不属于药监局批准药物，且二聚化生物元件使用的蛋白配体不具有生物正交性，其人体安全性有待考证。本研究项目拟以药监局批准的药物Danoprevir和丙肝病毒NS3/4A蛋白酶复合物为基础，运用Rosetta程序从头设计具有生物正交性的蛋白配体，并进一步定向进化成小分子化合物介导的异源二聚化的蛋白质元件，以用于下一代CAR-T细胞疗法。

Synthetic biology has been used in new therapeutic modalities, such as chimeric antigen receptor T (CAR-T) cell therapy, which has been successfully used to treat acute lymphoblastic leukemia and lymphomas. However, once the engineered CAR-T cells are injected into the patient, they cannot be effectively controlled, resulting in severe toxicity and even fatality. The use of small-molecule drugs as a switch to control the cell function can achieve temporal and spatial accuracy to regulate the function of engineered CAR-T cells in order to solve the safety issue of CAR-T cell therapy. Recently, some studies regulate the switch control of CAR-T cells by chemically induced dimerization tools. However, these small molecules are not FDA-approved drugs, and the proteins used in the chemically induced dimerization tools are not bio-orthogonal, so their human safeties are yet to be tested. This proposed project is using the Rosetta program to de novo design bio-orthogonal protein binder for the FDA-approved drug Danoprevir and HCV NS3/4A protease complex. Directed evolution will be further used to develop it into a chemically induced protein hetero-dimerization tool. The results could be used in next-generation CAR-T cell therapy.