開發用於降解或成像 SARS-CoV-2 主要蛋白酶的抑制型雙功能結合物

Abstract

SARS-CoV-2 (嚴重急性呼吸道症候群冠狀病毒 2) 是一種 RNA 病毒，在過去幾年間造成 COVID-19 全球大流行。主蛋白酶 (Mpro)，又稱為 3CL 蛋白酶，是 SARS-CoV-2 基因組中一個有價值的藥物靶點，對其病毒複製至關重要。在本專案中，我們希望將已知的單功能肽醛抑制劑升級，加入額外的生物功能，如成像和降解，同時減少生物系統中醛的缺點。首先評估了 N 端或 C 端延伸，以加入可點選的炔烴把手進行官能化。對於 N 端延伸，保留了醛彈頭。在 C 端延伸時，則以不可逆共價丙烯酰胺或可逆共價氰基丙烯酰胺取代醛。透過銅(I)催化的疊氮烷基環加成 (CuAAC)，螢光團或 E3-連接酵素招引物可以附著在抑制劑上。我們成功合成了 N 端和 C 端降解結合物，這些結合物保留了與 Mpro 的抑制作用，有些化合物在轉染細胞模型中顯示出 Mpro 的標記或降解，同時也抑制了病毒感染。

SARS-CoV-2, severe acute respiratory syndrome coronavirus 2, is the RNA virus that caused the COVID-19 global pandemic in the last few years. Main Protease (Mpro), also known as 3CL protease is a valuable drug target within the SARS-CoV-2 genome crucial for its viral replication. In this project, we would like to upgrade known monofunctional peptide aldehyde inhibitors to include extra biological functionalities, such as imaging and degradation, while reducing the drawbacks of aldehydes in biological systems. N- or C-terminal extension was first evaluated to include a clickable alkyne handle for functionalization. For N-terminal extension, the aldehyde warhead was retained. On the other hand, for C-terminal extensions, the aldehyde was replaced with irreversible covalent acrylamide or reversible covalent cyanoacrylamide moieties. Through copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC), fluorophores or E3-ligase recruiters can be attached to the inhibitor. We successfully synthesized N- and C-terminal degradation conjugates which retained their inhibition with Mpro, with some compounds showing labeling or degradation of Mpro in the transfection cell model, and also inhibition of viral infection.