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Antiviral Res . Antiviral Mechanisms and Preclinical Evaluation of Amantadine Analogs that Continue to Inhibit Influenza A Viruses with M2S31N-Based

tetano

Editor, Senior Moderator
Antiviral Res


. 2025 Feb 11:106104.
doi: 10.1016/j.antiviral.2025.106104. Online ahead of print. Antiviral Mechanisms and Preclinical Evaluation of Amantadine Analogs that Continue to Inhibit Influenza A Viruses with M2[SUP]S31N[/SUP]-Based Drug Resistance

Ian Tietjen[SUP] 1 [/SUP], Daniel C Kwan[SUP] 2 [/SUP], Annett Petrich[SUP] 3 [/SUP], Roland Zell[SUP] 4 [/SUP], Ivi Theodosia Antoniadou[SUP] 5 [/SUP], Agni Gavriilidou[SUP] 6 [/SUP], Christina Tzitzoglaki[SUP] 6 [/SUP], Michail Rallis[SUP] 7 [/SUP], David Fedida[SUP] 2 [/SUP], Francesc X Sureda[SUP] 8 [/SUP], Cato Mestdagh[SUP] 9 [/SUP], Lieve Naesens[SUP] 9 [/SUP], Salvatore Chiantia[SUP] 10 [/SUP], F Brent Johnson[SUP] 11 [/SUP], Antonios Kolocouris[SUP] 12 [/SUP]



Affiliations
Abstract

To better manage seasonal and pandemic influenza infections, new drugs are needed with enhanced activity against amantadine- and rimantadine-resistant influenza A virus (IAV) strains containing the S31N variant of the viral M2 ion channel (M2[SUP]S31N[/SUP]). Here we tested 36 amantadine analogs against a panel of viruses containing either M2[SUP]S31N[/SUP] or the parental, M2 S31 wild-type variant (M2[SUP]WT[/SUP]). We found that several analogs, primarily those with sizeable lipophilic adducts, inhibited up to three M2[SUP]S31N[/SUP]-containing viruses with activities at least 5-fold lower than rimantadine, without inhibiting M2[SUP]S31N[/SUP] proton currents or modulating endosomal pH. While M2[SUP]WT[/SUP] viruses in passaging studies rapidly gained resistance to these analogs through the established M2 mutations V27A and/or A30T, resistance development was markedly slower for M2[SUP]S31N[/SUP] viruses and did not associate with additional M2 mutations. Instead, a subset of analogs, exemplified by 2-propyl-2-adamantanamine (38), but not 2-(1-adamantyl)piperidine (26), spiro[adamantane-2,2'-pyrrolidine] (49), or spiro[adamantane-2,2'-piperidine] (60), inhibited cellular entry of infectious IAV following pre-treatment and/or H1N1 pseudovirus entry. Conversely, an overlapping subset of the most lipophilic analogs including compounds 26, 49, 60, and others, disrupted viral M2-M1 protein colocalization required for intracellular viral assembly and budding. Finally, a pilot toxicity study in mice demonstrated that 38 and 49 were tolerated at 30 mg/kg. Together, these results indicate that amantadine analogs act on multiple, complementary mechanisms to inhibit replication of M2[SUP]S31N[/SUP] viruses.

Keywords: Influenza A virus; M2 protein; amantadine analogs; antiviral mechanisms of action; drug resistance.

 
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