從新冠病毒的3C樣蛋白酶抑制劑來發現71型腸病毒的3C蛋白酶抑制劑

Abstract

腸病毒71型（EV71）曾在台灣引發多次疫情，過去造成數千名兒童死亡，目前仍無有效藥物治療。在EV71中，3C蛋白酶（3Cpro）是一種高度保守的半胱氨酸蛋白酶，具有非典型的Cys-His-Glu催化三聯體，對病毒複製過程中的多蛋白加工至關重要。SARS-CoV-2中的3C樣蛋白酶（3CLpro）負責將病毒多蛋白裂解為成熟的非結構蛋白（NSPs）。3CLpro因其結構類似於小RNA病毒的3C蛋白酶（3Cpro）而得名。在本論文中，我們旨在從已知的3CLpro抑制劑中識別3Cpro抑制劑，因此在本研究中，我首先建立3C蛋白酶的活性量測系統來篩選3C樣蛋白酶的抑制劑及測量他們對3C樣蛋白酶及3C蛋白酶的半抑制常數。 然而，篩選出一些含鋅配合物的胜肽類似抑制劑能同時有效抑制3CLpro和3Cpro。最後，電腦模擬用來釐清他們的結合樣式。

Enterovirus 71 (EV71) that caused outbreaks in Taiwan and killed thousands of children in the past, and currently, there is no effective drug against it. In EV71, 3C protease (3Cpro) is a highly conserved cysteine protease, featuring a non-canonical Cys-His-Glu catalytic triad essential for polyprotein processing during virus replication. The 3C-like protease (3CLpro) in SARS-CoV-2 is responsible to cleave the viral polyproteins into mature non-structural protein (NSPs). 3CLpro got its name as analogous to the picornavirus 3C protease (3Cpro) in structures. In this thesis, we aimed to identify 3Cpro inhibitors from the known 3CLpro inhibitors. I set up assay for 3Cpro to screen known 3CLpro inhibitors and measured their IC50 values against 3CLpro and 3Cpro. However, I screen out some peptide-like inhibitors with Zinc complexes that could effectively inhibit both 3CLpro and 3Cpro. Finally, computer modeling was used to rationalized their binding modes