Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems , Institute of Chemistry, Chinese Academy of Sciences (CAS) , Beijing 100190 , China.
University of Chinese Academy of Sciences , Beijing 100049 , China.
J Am Chem Soc. 2019 Nov 13;141(45):18136-18141. doi: 10.1021/jacs.9b08669. Epub 2019 Oct 17.
The selective and temporal control of protein activity in living cells provides a powerful tool to manipulate cellular function and to develop pro-protein therapeutics (PPT) for targeted therapy. In this work, we reported a facile but general chemical approach to design PPT by modulating protein activity in response to endogenous enzyme of disease cells, and its potential for targeted cancer therapy. We demonstrated that the chemical modification of a protein with quinone propionic acid (QPN), a ligand that could be reduced by tumor-cell-specific NAD(P)H dehydrogenase [quinone] 1 (NQO1), was reversible in the presence of NQO1. Importantly, the QPN-modified cytochrome c (Cyt c-QPN) and ribonuclease A (RNase A-QPN) showed NQO1-regulated protein activity in a highly selective manner. Furthermore, the intracellular delivery of RNase A-QPN using a novel type of lipid-based nanoparticles, and subsequent protein activation by cellular NQO1, selectively inhibit cancer cell growth in vitro and effectively suppress tumor growth in vivo. We believe that our approach increases the number of potentially useful chemical tools for reversibly controlling the structure and function of protein using a disease-cell-specific enzyme, opening opportunities in the study of dynamic biological processes and developing precise protein therapeutics.
在活细胞中选择性和时间控制蛋白质活性为操纵细胞功能和开发针对特定治疗的前体药物（PPT）提供了强大的工具。在这项工作中，我们报告了一种通过调节蛋白质活性来响应疾病细胞内源性酶来设计 PPT 的简便但通用的化学方法，及其在靶向癌症治疗中的潜力。我们证明了用醌丙酸（QPN）修饰蛋白质的化学方法，QPN 是一种可以被肿瘤细胞特异性 NAD（P）H 脱氢酶[醌]1（NQO1）还原的配体，在存在 NQO1 的情况下是可逆的。重要的是，QPN 修饰的细胞色素 c（Cyt c-QPN）和核糖核酸酶 A（RNase A-QPN）以高度选择性的方式表现出 NQO1 调节的蛋白质活性。此外，使用新型脂质纳米粒进行 RNase A-QPN 的细胞内递送，然后通过细胞 NQO1 进行蛋白质激活，可选择性地抑制体外癌细胞生长，并有效地抑制体内肿瘤生长。我们相信，我们的方法增加了使用疾病细胞特异性酶可逆控制蛋白质结构和功能的潜在有用化学工具的数量，为动态生物过程的研究和开发精确的蛋白质治疗开辟了机会。
