Oseltamivir aziridines are potent influenza neuraminidase inhibitors and imaging agents
Abstract
Influenza neuraminidase (NA) is a critical target for seasonal and pandemic antivirals, including the strains of current concern. Current treatments, such as Zanamivir and Oseltamivir, are limited by noncovalent binding and emerging resistance. We hypothesized that Oseltamivir aziridines would unite transition-state mimicry for tight binding, with aziridine-enabled covalent capture of the catalytic tyrosine, thereby supporting both therapy and activity-based quantification. Here, we present oseltamivir-based aziridines, inspired by cyclophellitol chemistry, that act as covalent inhibitors and activity-based probes via an N -acylaziridine warhead. Free-energy calculations, and NMR observations, indicate a 4 H 5 half-chair preference consistent with the NA transition state, and selected analogues inhibit multiple NA subtypes with low nanomolar binding constants. Diverse evidence establishes covalency: time-dependent inactivation, inhibitor washout, intact-mass shifts, MS/MS identification of a tyrosine adduct, and QM/MM reaction profiles, while cryoEM of N1 aligns with the proposed binding mode, revealing an elimination product. The inhibitors demonstrate formidable activity against diverse viral neuraminidases, including H5N1, and further enable imaging and quantification of active NA. With their dual therapeutic and diagnostic potential, these first-in-class inhibitors indeed benefit from transition state mimicry and covalency, and thus offer a powerful platform for antiviral development and neuraminidase imaging, addressing urgent global health needs in influenza treatment and prevention.
Article Details
Journal Info
Proceedings of the National Academy of Sciences
National Academy of Sciences
Authors (23)
Merijn B. L. Vriends
Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University
Elisha Moran
Department of Chemistry, University of York Heslington
Martín Calvelo
Centro Singular de Investigación en Química Biolóxica e Materiais Moleculares (CiQUS) and Departamento de Química Orgánica Universidade de Santiago de Compostela
Thomas Hansen
Isabelle B. Pickles
Department of Chemistry, The University York, Heslington, York YO10 5DD, United Kingdom
Xincheng Xin
Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University
Marieke Biezeno
Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University
Zachary W. B. Armstrong
Leiden Institute of Chemistry, Leiden University
Maria J. Ferraz
Department of Medical Biochemistry, Leiden Institute of Chemistry, Leiden University
Lei Li
Alice Lilley
Worldwide Influenza Centre, The Francis Crick Institute
Ruth Harvey
Dmitri V. Filippov
Leiden Institute of Chemistry, Leiden University , , ,
Qinghua Liao
Departament de Química Inorgaǹica i Orgaǹica (Seccióde Química Orgaǹica) and Institut de Química TeorÌica I Computacional, Universitat de Barcelona
Sybrin P. Schröder
Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University
Gijsbert A. van der Marel
Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands
Marta Artola
Department of Medical Biochemistry, Leiden Institute of Chemistry, Leiden University
Johannes M. F. G. Aerts
Department of Medical Biochemistry, Leiden Institute of Chemistry, Leiden University
James N. Blaza
Jeroen D. C. Codée
Leiden Institute of Chemistry
Carme Rovira
Departament de Química Inorgànica i Orgànica (Secció de Química Orgànica) and Institut de Química Teòrica i Computacional (IQTCUB)
Herman S. Overkleeft
Leiden Institute of Chemistry
Gideon J. Davies