以核糖核酸酶P的RNA组分为作用靶标的抗结核反义寡核苷酸的研究

In recent years, the emergence of multi-drug resistant tuberculosis (TB) has been continuingly increased. The discovery of antitubercular drugs with fully new antibacterial mechanisms is quite demanding. RNase P catalyses the 5'-maturation of tRNA, and is an essential enzyme in almost all organisms. There are large differences in structure and function between bacterial and human RNase P RNA subunits, and therefore the bacterial RNase P RNA is believed to be an ideal antibacterial drug target. This project plans to screen antisense oligonucleotides that target M. tuberculosis RNase P RNA and can effectively inhibit or cease the growth of Mycobacterium tuberculosis by utilizing rational design, affinity assays of macromolecules, kinetic assays, BCG antibacterial activity screening, and new antisense technologies. The project is to identify the targeting sites of antisense oligonucleotides on M. tuberculosis RNase P RNA, and characterize the mechanisms of the antisense oligonucleotides with antibacterial activity. The goal of the project is to contribute to the discovery of new antisense oligonucleotide drug candidates against multi-drug resistant TB.

近年来耐多药结核病的发病率呈上升趋势，人类迫切需要具有全新作用机制的抗结核药物。核糖核酸酶P是存在于几乎所有的生物体中的催化tRNA 5'端成熟的必需酶。细菌与人的核糖核酸酶P的 RNA组分在结构和功能上有巨大差异，是理想的抗菌药物作用靶标。本项目拟以结核分枝杆菌的核糖核酸酶P RNA为作用靶标，通过合理设计、生物大分子间亲和力研究、酶动力学研究和卡介苗（BCG）抗菌活性筛选等手段，并结合新一代的反义技术，筛选能有效阻截靶RNA功能，从而抑制甚至竭制结核分枝杆菌生长和繁殖的反义寡核苷酸。揭示结核分枝杆菌核糖核酸酶P RNA结构中的反义寡核苷酸作用位点，探索具有抗菌活性的反义寡核苷酸的作用机制。该项目可望为开发有效治疗和对抗耐多药结核病的反义寡核苷酸药物奠定坚实基础。

近年来耐多药结核病的发病率呈上升趋势，人类迫切需要具有全新作用机制的抗结核药物。核糖核酸酶P是存在于几乎所有的生物体中的催化tRNA 5'端成熟的必需酶。细菌与人的核糖核酸酶P的 RNA组分在结构和功能上有巨大差异，是理想的抗菌药物作用靶标。本项目以结核分枝杆菌的核糖核酸酶P RNA为作用靶标，通过合理设计、荧光定量生物大分子间亲和力研究和表达eGFP的卡介苗（BCG）抗菌活性筛选等手段，筛选能有效阻截靶RNA功能，从而抑制甚至竭制结核分枝杆菌生长和繁殖的反义寡核苷酸，确证和揭示了结核分枝杆菌核糖核酸酶P RNA结构中的精准抗菌关键作用位点。该项目可望为开发有效治疗和对抗耐多药结核病的反义寡核苷酸药物奠定坚实基础。