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Investigation of the Drug Resistance Mechanism of M2-S31N Channel Blockers through Biomolecular Simulations and Viral Passage Experiments
journal contribution
posted on 2020-04-10, 17:14 authored by Rami Musharrafieh, Panagiotis Lagarias, Chunlong Ma, Raymond Hau, Alex Romano, George Lambrinidis, Antonios Kolocouris, Jun WangRecent efforts in drug development
against influenza A virus (IAV)
M2 proton channel S31N mutant resulted in conjugates of amantadine
linked with aryl head heterocycles. To understand the mechanism of
drug resistance, we chose a representative M2-S31N inhibitor, compound 3, as a chemical probe to identify resistant mutants. To increase
the possibility of identifying novel resistant mutants, serial viral
passage experiments were performed with multiple strains of H1N1 and
H3N2 viruses in different cell lines. This approach not only identified
M2 mutations around the drug-binding site, including the pore-lining
residues (V27A, V27F, N31S, and G34E) and an interhelical residue
(I32N), but also a new allosteric mutation (R45H), in addition to
L46P previously identified, located at the C-terminus of M2 that is
more than 10 Å away from the drug-binding site. The effects of
each mutation were next investigated using electrophysiology, recombinant
viruses, and molecular dynamics (MD) simulations. The reduced sensitivity
in channel blockage correlated with increased drug resistance in antiviral
assays using recombinant viruses. The MD simulations show that the
V27A, V27F, G34E, and R45H mutations increase the diameter and hydration
state of the pore in complex with compound 3. The Molecular
Mechanics Generalized Born (MM-GBSA) calculations result in more positive
binding free energies for the complexes of resistant M2 (V27A, V27F,
G34E, R45H) with compound 3 compared to the stable complexes
(S31N and I32N). Overall, this is the first systematic study of the
drug resistance mechanism of M2-S31N channel blockers using multiple
viruses in different cell lines.
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compound 3M 2-S Channel Blockersaryl head heterocyclesR 45H mutations increaseH 3N virusesIAVcell linesV 27A V 27F G 34EM 2-S channel blockersresidueMD simulations showV 27A V 27F G 34E R 45Hrepresentative M 2-S inhibitorMM-GBSA32NM 2 mutations46Pdrug-binding siteM 2 proton channel S 31NV 27A V 27F N 31SMolecular Mechanics GeneralizedDrug Resistance MechanismViral Passage ExperimentsM 2drug resistancedrug resistance mechanismmutantcomplex
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