biostudies-literaturePresley LNIAID NIH HHS2616944https://www.ebi.ac.uk/biostudies/studies/S-EPMC12862872biostudies-literatureUnknown15(1)Human TMPRSS2 is a type II transmembrane serine protease and an essential host factor for SARS-CoV-2 and influenza A virus (IAV H1N1) infections. It facilitates the cleavage of viral surface glycoproteins, which are required for membrane fusion. This importance makes it an attractive target for host-directed antiviral therapies. We previously identified N-0385 and N-0920 as nanomolar TMPRSS2 inhibitors and demonstrated their antiviral potency against several SARS-CoV-2 variants. Here, we screened another twelve N-0385/N-0920 analogs with improved pharmacokinetics. Compounds <b>9</b> and <b>10</b> showed strong inhibition of TMPRSS2 activity and viral entry: they blocked pseudoviruses and authentic SARS-CoV-2 JN.1 and IAV H1N1 in Calu-3 cells. Compound <b>9</b> displayed a synergistic effecEmerging microbes & infectionsTMPRSS2 inhibitors with broad-spectrum efficacy against SARS-CoV-2 (JN.1) and influenza A (H1N1) viruses protect mice from influenza A infection.PMC12862872DP1 AI158186P01 AI16796675N93022C00036Presley LSaintigny AEnnis SLeduc RBrinsden LVillanueva IChen MFone JFerkova SVeesler DBrown JTJean FBoudreault PLMcCallum MNiikura MPerez-Vargas JHo MThompson CAHKaushik NBautista-Sanchez DWarner BOlver SHetrick BfalseTMPRSS2 inhibitors with broad-spectrum efficacy against SARS-CoV-2 (JN.1) and influenza A (H1N1) viruses protect mice from influenza A infection.Human TMPRSS2 is a type II transmembrane serine protease and an essential host factor for SARS-CoV-2 and influenza A virus (IAV H1N1) infections. It facilitates the cleavage of viral surface glycoproteins, which are required for membrane fusion. This importance makes it an attractive target for host-directed antiviral therapies. We previously identified N-0385 and N-0920 as nanomolar TMPRSS2 inhibitors and demonstrated their antiviral potency against several SARS-CoV-2 variants. Here, we screened another twelve N-0385/N-0920 analogs with improved pharmacokinetics. Compounds <b>9</b> and <b>10</b> showed strong inhibition of TMPRSS2 activity and viral entry: they blocked pseudoviruses and authentic SARS-CoV-2 JN.1 and IAV H1N1 in Calu-3 cells. Compound <b>9</b> displayed a synergistic effec2026-01-01T00:00:00Z2026 Dec2026-07-14T21:27:13.233Z2026-06-25T03:08:01.869ZS-EPMC128628724161288710.1080/22221751.2026.2616944