Kiew Lik-Voon, Chang Chia-Yu, Huang Sheng-Yu, Wang Pei-Wen, Heh Choon-Han, Liu Chung-Te, Cheng Chia-Hsin, Lu Yi-Xiang, Chen Yen-Chen, Huang Yi-Xuan, Chang Sheng-Yun, Tsai Huei-Yu, Kung Yu-An, Huang Peng-Nien, Hsu Ming-Hua, Leo Bey-Fen, Foo Yiing-Yee, Su Chien-Hao, Hsu Kuo-Chen, Huang Po-Hsun, Ng Chirk-Jenn, Kamarulzaman Adeeba, Yuan Chiun-Jye, Shieh Dar-Bin, Shih Shin-Ru, Chung Lip-Yong, Chang Chia-Ching
Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia; Department of Biological Science and Technology, College of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsinchu, 30068, Taiwan.
Department of Biological Science and Technology, College of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsinchu, 30068, Taiwan; Center for Intelligent Drug Systems and Smart Bio-devices (IDS(2)B), National Yang Ming Chiao Tung University, Hsinchu, 30068, Taiwan.
Biosens Bioelectron. 2021 Jul 1;183:113213. doi: 10.1016/j.bios.2021.113213. Epub 2021 Apr 3.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enters the cells through the binding of its spike protein (S-protein) to the cell surface-expressing angiotensin-converting enzyme 2 (ACE2). Thus, inhibition of S-protein-ACE2 binding may impede SARS-CoV-2 cell entry and attenuate the progression of Coronavirus disease 2019 (COVID-19). In this study, an electrochemical impedance spectroscopy-based biosensing platform consisting of a recombinant ACE2-coated palladium nano-thin-film electrode as the core sensing element was fabricated for the screening of potential inhibitors against S-protein-ACE2 binding. The platform could detect interference of small analytes against S-protein-ACE2 binding at low analyte concentration and small volume (0.1 μg/mL and ~1 μL, estimated total analyte consumption < 4 pg) within 21 min. Thus, a few potential inhibitors of S-protein-ACE2 binding were identified. This includes (2S,3aS,6aS)-1-((S)-N-((S)-1-Carboxy-3-phenylpropyl)alanyl)tetrahydrocyclopenta[b] pyrrole-2-carboxylic acid (ramiprilat) and (2S,3aS,7aS)-1-[(2S)-2-[[(2S)-1-Carboxybutyl]amino]propanoyl]-2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (perindoprilat) that reduced the binding affinity of S-protein to ACE2 by 72% and 67%; and SARS-CoV-2 in vitro infectivity to the ACE2-expressing human oral cavity squamous carcinoma cells (OEC-M1) by 36.4 and 20.1%, respectively, compared to the PBS control. These findings demonstrated the usefulness of the developed biosensing platform for the rapid screening of modulators for S-protein-ACE2 binding.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）通过其刺突蛋白（S蛋白）与细胞表面表达的血管紧张素转换酶2（ACE2）结合进入细胞。因此，抑制S蛋白与ACE2的结合可能会阻碍SARS-CoV-2进入细胞，并减缓2019冠状病毒病（COVID-19）的进展。在本研究中，构建了一种基于电化学阻抗谱的生物传感平台，该平台以重组ACE2包被的钯纳米薄膜电极作为核心传感元件，用于筛选针对S蛋白与ACE2结合的潜在抑制剂。该平台能够在低分析物浓度和小体积（0.1μg/mL和约1μL，估计总分析物消耗量<4pg）下，在21分钟内检测小分子分析物对S蛋白与ACE2结合的干扰。因此，鉴定出了几种S蛋白与ACE2结合的潜在抑制剂。其中包括（2S，3aS，6aS）-1-（（S）-N-（（S）-1-羧基-3-苯基丙基）丙氨酰基）四氢环戊[b]吡咯-2-羧酸（雷米普利拉）和（2S，3aS，7aS）-1-[(2S)-2-[[(2S)-1-羧基丁基]氨基]丙酰基]-2,3,3a,4,5,6,7,7a-八氢吲哚-2-羧酸（培哚普利拉），它们分别使S蛋白与ACE2的结合亲和力降低了72%和67%；与PBS对照相比，SARS-CoV-2对表达ACE2的人口腔鳞状癌细胞（OEC-M1）的体外感染性分别降低了36.4%和20.1%。这些发现证明了所开发的生物传感平台在快速筛选S蛋白与ACE2结合调节剂方面的实用性。
