新冠病毒在國際間快速的傳播及不斷的變異，已經成為了全球嚴重的公共衛生負擔，除了疫苗外我們還需要找到另一個有效預防或降低新冠病毒感染風險的方式。在小分子藥物的開發中，目前有Remdesivir、Favipiravir、Molnupiravir、Paxlovid等少數已核可藥物用於治療此疾病。因此本篇研究目的即是找到能抑制新冠病毒感染人類細胞的活性化合物，以利於後續抗新冠病毒藥物的開發。由於新冠病毒附著於人類細胞受體屬於RBD-ACE2蛋白質間交互作用力，藉由篩選1068種美國藥物食品管理局核准的藥物，已有文獻報導Ivermectin、Gramicidin巨環類藥物對於新冠病毒RBD附著於人類細胞受體ACE2有抑制效果。亦研究也指出巨環類化合物可以當作蛋白質間交互作用力抑制劑，因此我們認為巨環類化合物很有機會成為阻斷新冠病毒感染的活性化合物。本篇利用RBD-ACE2 Attachment Assay、Pseudovirus Neutralization Assay、Cytotoxicity從2560個不同的巨環類化合物中成功地篩選出1個全新的結構分子Mac734，對於人類細胞不具細胞毒性且擁有抑制新冠病毒主流變異株感染的效果，分別對Alpha (B.1.1.7)、Beta (B.1.351)、Delta (B.1.671.2)、Omicron (B.1.1.529)附著到人類細胞受體的EC50依序為22.2 M、10.3 M、9.5 M、5.4 M。未來可以根據此分子進行官能基的修飾，以利於抗新冠病毒藥物的開發。此外根據Mac734的結構，利用點擊化學快速合成一系列的結構類似物，且發現化合物5g能促進不同新冠病毒主流變異株感染人類細胞的效果。綜觀以上結果，本篇目前各別發現一個新冠病毒抑制劑與促進劑，日後皆有潛力成為研究抗新冠病毒感染的利器。

　Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has become a global public health burden. In addition to vaccines, effective prevention and reduction of SARS-CoV-2 associated risks are desired. Current development of small molecule drugs has yielded approved drugs such as Remdesivir, Favipiravir, Molnupiravir, and Paxlovid for the treatment of SARS-CoV-2. The purpose of this study is to identify new protein-protein interaction (PPI) inhibitor in the attachment of human cell receptor Angiotensin-Converting Enzyme 2 (ACE2) by SARS-CoV-2 Receptor Binding Domain (RBD) on the spike protein. By screening of 1068 FDA-approved drugs, previous study has identified macrocyclic drugs, such as Ivermectin and Gramicidin to exert an inhibitory effect on the attachment between RBD and ACE2. Moreover, numerous reports have leveraged macrocyclic scaffolds as PPI inhibitors. Herein, we used RBD-ACE2 attachment assay, pseudovirus neutralization assay, host cell cytotoxicity assay to successfully identify a new macrocyclic molecule Mac734. By click chemistry, we also synthesized a series of structural analogs, and surprisingly, found that analog 5g could promote the entry of SARS-CoV-2 variants during pseudovirus infection. In summary, this study has disclosed a new SARS-CoV-2 entry inhibitor and an enhancer, both of which are potential powerful tools for research on coronavirus pathogenesis.

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