{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Starvaggi J"],"funding":["PRIN_2022PNRR"],"pagination":["24"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC12845239"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["19(1)"],"pubmed_abstract":["<b>Background:</b> In this study, we report a novel series of proline- and pipecolic acid-based small molecules designed as allosteric inhibitors of the NS2B/NS3 serine proteases from dengue and Zika viruses, key targets in antiviral drug discovery. <b>Results:</b> Enzymatic studies revealed that <i>S</i>-proline derivatives bearing electron-withdrawing substituents on the aromatic ring, particularly that with a trifluoromethyl group in <i>meta</i> position (i.e., compound <b>3</b>, IC<sub>50</sub> = 5.0 µM), were the most potent against DENV NS2B/NS3, while nitro-substituted inhibitors were mostly effective only against the ZIKV protease. <i>R</i>-configured pipecolic acid-based derivatives were the only ones active against DENV NS2B/NS3, even if the mid-micromolar range; however, they demonstrated improved cellular efficacy since inhibitors <b>24</b> and <b>27</b> exhibiting strong activity in a DENV2 protease reporter gene assay (EC<sub>50</sub> = 5.2 and 5.1 µM, respectively). All compounds showed no cytotoxicity (CC<sub>50</sub> > 100 µM) and were selective for the viral protease over off-target serine proteases. Structure-based approaches were exploited to map the druggable allosteric site close to Asn152. <b>Conclusions:</b> Our findings led us to identify proline and pipecolic acid-based inhibitors as promising leads for the development of selective flaviviral NS2B/NS3 allosteric inhibitors."],"journal":["Pharmaceuticals (Basel, Switzerland)"],"pubmed_title":["Development of Novel Proline- and Pipecolic Acid-Based Allosteric Inhibitors of Dengue and Zika Virus NS2B/NS3 Protease."],"pmcid":["PMC12845239"],"funding_grant_id":["P2022KCW3LP2022KCW3L_001 and P2022KCW3LP2022KCW3L_002"],"pubmed_authors":["Lang J","Klein C","Starvaggi J","Belgiovine V","Nicolotti O","Di Chio C","Ettari R","Previti S","Trisciuzzi D","Di Maro S","Zappala M"],"additional_accession":[]},"is_claimable":false,"name":"Development of Novel Proline- and Pipecolic Acid-Based Allosteric Inhibitors of Dengue and Zika Virus NS2B/NS3 Protease.","description":"<b>Background:</b> In this study, we report a novel series of proline- and pipecolic acid-based small molecules designed as allosteric inhibitors of the NS2B/NS3 serine proteases from dengue and Zika viruses, key targets in antiviral drug discovery. <b>Results:</b> Enzymatic studies revealed that <i>S</i>-proline derivatives bearing electron-withdrawing substituents on the aromatic ring, particularly that with a trifluoromethyl group in <i>meta</i> position (i.e., compound <b>3</b>, IC<sub>50</sub> = 5.0 µM), were the most potent against DENV NS2B/NS3, while nitro-substituted inhibitors were mostly effective only against the ZIKV protease. <i>R</i>-configured pipecolic acid-based derivatives were the only ones active against DENV NS2B/NS3, even if the mid-micromolar range; however, they demonstrated improved cellular efficacy since inhibitors <b>24</b> and <b>27</b> exhibiting strong activity in a DENV2 protease reporter gene assay (EC<sub>50</sub> = 5.2 and 5.1 µM, respectively). All compounds showed no cytotoxicity (CC<sub>50</sub> > 100 µM) and were selective for the viral protease over off-target serine proteases. Structure-based approaches were exploited to map the druggable allosteric site close to Asn152. <b>Conclusions:</b> Our findings led us to identify proline and pipecolic acid-based inhibitors as promising leads for the development of selective flaviviral NS2B/NS3 allosteric inhibitors.","dates":{"release":"2025-01-01T00:00:00Z","publication":"2025 Dec","modification":"2026-06-13T03:19:47.754Z","creation":"2026-06-13T03:12:01.199Z"},"accession":"S-EPMC12845239","cross_references":{"pubmed":["41599626"],"doi":["10.3390/ph19010024"]}}