Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Life Sciences Center, Vilnius University, Saulėtekio 7, LT-10257 Vilnius, Lithuania.
Department of Microbiology and Biotechnology, Institute of Biosciences, Life Sciences Center, Vilnius University, Saulėtekio 7, LT-10257 Vilnius, Lithuania.
Int J Mol Sci. 2023 Aug 30;24(17):13491. doi: 10.3390/ijms241713491.
The severe acute respiratory syndrome-causing coronavirus 2 (SARS-CoV-2) papain-like protease (PL) and main protease (M) play an important role in viral replication events and are important targets for anti-coronavirus drug discovery. In search of these protease inhibitors, we screened a library of 1300 compounds using a fluorescence thermal shift assay (FTSA) and identified 53 hits that thermally stabilized or destabilized PL. The hit compounds structurally belonged to two classes of small molecules: thiazole derivatives and symmetrical disulfide compounds. Compound dissociation constants (K) were determined using an enzymatic inhibition method. Seven aromatic disulfide compounds were identified as efficient PL inhibitors with K values in the micromolar range. Two disulfides displayed six-fold higher potency for PL (K = 0.5 µM) than for M. The disulfide derivatives bound covalently to both proteases, as confirmed through mass spectrometry. The identified compounds can serve as lead compounds for further chemical optimization toward anti-COVID-19 drugs.
严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）木瓜蛋白酶样蛋白酶（PL）和主要蛋白酶（M）在病毒复制事件中发挥重要作用，是抗冠状病毒药物发现的重要靶点。为了寻找这些蛋白酶抑制剂，我们使用荧光热转移分析（FTSA）筛选了 1300 种化合物库，鉴定出 53 种能够热稳定或不稳定 PL 的化合物。这些命中化合物在结构上属于两类小分子：噻唑衍生物和对称二硫化物化合物。使用酶抑制方法测定化合物的解离常数（K）。确定了 7 种芳香族二硫化物化合物是有效的 PL 抑制剂，其 K 值在微摩尔范围内。两种二硫化物对 PL（K=0.5µM）的效力比 M 高六倍。通过质谱证实，二硫化物衍生物与两种蛋白酶均以共价键结合。鉴定出的化合物可作为进一步化学优化以开发抗 COVID-19 药物的先导化合物。
