Wei Chao, Li Yuhua, Guo Lina, Shao Zhiyu, Diao Hua
College of Chemistry and Chemical Engineering, Donghua University, Shanghai 201620, China.
NHC Key Lab of Reproduction Regulation, Shanghai Engineering Research Center of Reproductive Health Drug and Devices, Shanghai-MOST Key Laboratory of Health and Disease Genomics, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Shanghai 200237, China.
Int J Mol Sci. 2025 Apr 21;26(8):3903. doi: 10.3390/ijms26083903.
3CL protease (3CL), a key enzyme of SARS-CoV-2 replication, is one of the most selective targets of antivirals, as no homologous protease has been recognized in the human body. As proteolysis-targeting chimeras (PROTACs) are superior to traditional inhibitors, based on the reported cereblon (CRBN) ligands thalidomide and lenalidomide, 3CL ligands of peptidomimetic inhibitors, and suitable linkers, we aimed to develop novel PROTACs that may trigger efficient intracellular 3CL degradation through a balance of hydrophilicity and lipophilicity. In brief, we designed and synthesized 5 PROTAC molecules. The 3CL degradation efficiency of the PROTACs was assayed in stable SARS-CoV-2 3CL expression HEK293 cell models and evaluated by Western blot. All compounds showed prominent 3CL degradation activity with tolerable HEK293 cytotoxicity. The most prominent PROTAC compounds, and , have DC values of approximately 1 µM, and D of 89.3% and 75% respectively, indicating good potential for further application.
3C样蛋白酶（3CL）是严重急性呼吸综合征冠状病毒2（SARS-CoV-2）复制的关键酶，由于人体中未发现同源蛋白酶，它是最具选择性的抗病毒靶点之一。鉴于蛋白酶靶向嵌合体（PROTAC）优于传统抑制剂，基于已报道的大脑神经酰胺（CRBN）配体沙利度胺和来那度胺、拟肽类抑制剂的3CL配体以及合适的连接子，我们旨在开发新型PROTAC，通过亲水性和疏水性的平衡引发有效的细胞内3CL降解。简而言之，我们设计并合成了5种PROTAC分子。在稳定表达SARS-CoV-2 3CL的HEK293细胞模型中测定PROTAC的3CL降解效率，并通过蛋白质印迹法进行评估。所有化合物均表现出显著的3CL降解活性，且对HEK293细胞具有可耐受的细胞毒性。最突出的PROTAC化合物和的DC值约为1 μM，D值分别为89.3%和75%，显示出良好的进一步应用潜力。
