Influenza A M2 Inhibitor Binding Understood through Mechanisms of Excess Proton Stabilization and Channel Dynamics

J Am Chem Soc. 2020 Oct 14;142(41):17425-17433. doi: 10.1021/jacs.0c06419. Epub 2020 Sep 29.

Abstract

Prevalent resistance to inhibitors that target the influenza A M2 proton channel has necessitated a continued drug design effort, supported by a sustained study of the mechanism of channel function and inhibition. Recent high-resolution X-ray crystal structures present the first opportunity to see how the adamantyl amine class of inhibitors bind to M2 and disrupt and interact with the channel's water network, providing insight into the critical properties that enable their effective inhibition in wild-type M2. In this work, we examine the hypothesis that these drugs act primarily as mechanism-based inhibitors by comparing hydrated excess proton stabilization during proton transport in M2 with the interactions revealed in the crystal structures, using the Multiscale Reactive Molecular Dynamics (MS-RMD) methodology. MS-RMD, unlike classical molecular dynamics, models the hydrated proton (hydronium-like cation) as a dynamic excess charge defect and allows bonds to break and form, capturing the intricate interactions between the hydrated excess proton, protein atoms, and water. Through this, we show that the ammonium group of the inhibitors is effectively positioned to take advantage of the channel's natural ability to stabilize an excess protonic charge and act as a hydronium mimic. Additionally, we show that the channel is especially stable in the drug binding region, highlighting the importance of this property for binding the adamantane group. Finally, we characterize an additional hinge point near Val27, which dynamically responds to charge and inhibitor binding. Altogether, this work further illuminates a dynamic understanding of the mechanism of drug inhibition in M2, grounded in the fundamental properties that enable the channel to transport and stabilize excess protons, with critical implications for future drug design efforts.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adamantane / chemistry
  • Amino Acid Sequence
  • Antiviral Agents / chemistry*
  • Binding Sites
  • Drug Design
  • Humans
  • Influenza A virus / metabolism*
  • Influenza, Human / metabolism*
  • Influenza, Human / prevention & control
  • Molecular Dynamics Simulation
  • Protein Binding
  • Protein Conformation
  • Protons
  • Viral Matrix Proteins / antagonists & inhibitors*
  • Viral Matrix Proteins / chemistry

Substances

  • Antiviral Agents
  • M2 protein, Influenza A virus
  • Protons
  • Viral Matrix Proteins
  • Adamantane