Structure-guided engineering of a mutation-tolerant inhibitor peptide against variable SARS-CoV-2 spikes

S Shun Nakamura Y Yukihiro Tanimura (Cellular and Structural Physiology Laboratory, Advanced Research Initiative, Institute of Integrated Research, Institute of Science Tokyo) R Risa Nomura (Cellular and Structural Physiology Laboratory, Advanced Research Initiative, Institute of Integrated Research, Institute of Science Tokyo) H Hiroshi Suzuki (Department of Polymer Chemistry, Graduate School of Engineering) K Kouki Nishikawa (CeSPIA Inc., Taisei Otemachi) A Akiko Kamegawa (Cellular and Structural Physiology Laboratory, Advanced Research Initiative, Institute of Integrated Research, Institute of Science Tokyo) N Nobutaka Numoto (International Center for Structural Biology, Research Institute for Interdisciplinary Science, Okayama University) A Atsushi Tanaka (Division of Research Animal Laboratory and Translational Medicine, Research and Development Center, Osaka Medical and Pharmaceutical University) S Shigeru Kawabata (Department of Pathology, Faculty of Medicine, Osaka Medical and Pharmaceutical University) S Shoichi Sakaguchi (Department of Microbiology and Infection Control, Faculty of Medicine, Osaka Medical and Pharmaceutical University) A Akino Emi (Department of Microbiology and Infection Control, Faculty of Medicine, Osaka Medical and Pharmaceutical University) Y Youichi Suzuki (Department of Microbiology and Infection Control, Faculty of Medicine, Osaka Medical and Pharmaceutical University) Y Yoshinori Fujiyoshi (Cellular and Structural Physiology Laboratory, Advanced Research Initiative, Institute of Integrated Research, Institute of Science Tokyo)

Abstract

Pathogen mutations present an inevitable and challenging problem for therapeutics and the development of mutation-tolerant anti-infective drugs to strengthen global health and combat evolving pathogens is urgently needed. While spike proteins on viral surfaces are attractive targets for preventing viral entry, they mutate frequently, making it difficult to develop effective therapeutics. Here, we used a structure-guided strategy to engineer an inhibitor peptide against the SARS-CoV-2 spike, called CeSPIACE, with mutation-tolerant and potent binding ability against all variants to enhance affinity for the invariant architecture of the receptor-binding domain (RBD). High-resolution structures of the peptide complexed with mutant RBDs revealed a mechanism of mutation-tolerant inhibition. CeSPIACE bound major mutant RBDs with picomolar affinity and inhibited infection by SARS-CoV-2 variants in VeroE6/TMPRSS2 cells (IC 50 4 pM to 13 nM) and demonstrated potent in vivo efficacy by inhalation administration in hamsters. Mutagenesis analyses to address mutation risks confirmed tolerance against existing and/or potential future mutations of the RBD. Our strategy of engineering mutation-tolerant inhibitors may be applicable to other infectious diseases.

Article Details

Volume / Issue Vol. 122, Issue 4
Published January 28, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (13)

S

Shun Nakamura

Y

Yukihiro Tanimura

Cellular and Structural Physiology Laboratory, Advanced Research Initiative, Institute of Integrated Research, Institute of Science Tokyo

R

Risa Nomura

Cellular and Structural Physiology Laboratory, Advanced Research Initiative, Institute of Integrated Research, Institute of Science Tokyo

H

Hiroshi Suzuki

Department of Polymer Chemistry, Graduate School of Engineering

K

Kouki Nishikawa

CeSPIA Inc., Taisei Otemachi

A

Akiko Kamegawa

Cellular and Structural Physiology Laboratory, Advanced Research Initiative, Institute of Integrated Research, Institute of Science Tokyo

N

Nobutaka Numoto

International Center for Structural Biology, Research Institute for Interdisciplinary Science, Okayama University

A

Atsushi Tanaka

Division of Research Animal Laboratory and Translational Medicine, Research and Development Center, Osaka Medical and Pharmaceutical University

S

Shigeru Kawabata

Department of Pathology, Faculty of Medicine, Osaka Medical and Pharmaceutical University

S

Shoichi Sakaguchi

Department of Microbiology and Infection Control, Faculty of Medicine, Osaka Medical and Pharmaceutical University

A

Akino Emi

Department of Microbiology and Infection Control, Faculty of Medicine, Osaka Medical and Pharmaceutical University

Y

Youichi Suzuki

Department of Microbiology and Infection Control, Faculty of Medicine, Osaka Medical and Pharmaceutical University

Y

Yoshinori Fujiyoshi

Cellular and Structural Physiology Laboratory, Advanced Research Initiative, Institute of Integrated Research, Institute of Science Tokyo