Gan Luo, Gang Wang, Chongbing Liao, Baisen Zhong, Lianfeng Fan, Jue Zhang, Jie Rao, Junjie Zhang, Ziqiao Wang, Man Luo, Yan YAN, Lu Lu, Wei Xu, Fan Wu, Xihui Gao, Wuyuan Lu
It remains elusive to design peptidomimetic inhibitors of SARS-CoV-2 main protease (Mpro) refractory to multiple deficiencies of Paxlovid (ritonavir-boosted nirmatrelvir), pertaining mainly to E166X mutations-conferred drug resistance and inherent pharmacokinetic limitations to nirmatrelvir. We identify via virtual screening an iso-quinoline P1 moiety in place of the traditional γ-lactam and design iso-quinoline-containing inhibitors with high affinity for Mpro and its nirmatrelvir-resistant E166X mutants. Further optimization at P4 cultivates distinctive peptidomimetic inhibitors with drastically improved pharmacokinetic properties and significantly enhanced antiviral efficacy independent of ritonavir. Two such inhibitors, FD3-32 and FD3-36, also potent against SARS-CoV-1 and MERS-CoV Mpro, are more effective as a monotherapy regimen than Paxlovid in reducing viral loads in vivo and protecting infected male mice from acute lung injury. Here, we report the discovery of next-generation SARS-CoV-2 Mpro inhibitors that overcome the deficiencies of Paxlovid, promising efficacious antivirals critical for mitigating the current and future pandemics of coronaviruses. Despite being a wonder drug for COVID-19, Paxlovid suffers deficiencies pertinent to ritonavir associated toxicity and rising resistance to nirmatrelvir, which authors in this work show can be overcome by redesigned viral main protease inhibitors as a mono-therapeutic agent.