{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T12:54:17Z","timestamp":1791982457039,"version":"4.4.0"},"reference-count":24,"publisher":"American Chemical Society (ACS)","issue":"40","license":[{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32272571"],"award-info":[{"award-number":["32272571"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32472593"],"award-info":[{"award-number":["32472593"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2022YFD1802102"],"award-info":[{"award-number":["2022YFD1802102"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Phenylpyrazole ectoparasiticides, exemplified by fipronil, act as noncompetitive antagonists of arthropod GABA receptors by blocking the ion channel pore. Here, we investigated the molecular mechanism of the newer phenylpyrazole nicofluprole using Drosophila melanogaster. CRISPR\/Cas9-mediated mutagenesis and pharmacological assays showed that A301 substitutions in the pore-forming TM2 helix conferred strong resistance to fipronil but had little effect on nicofluprole sensitivity. In contrast, G335 mutations in TM3 markedly reduced nicofluprole activity. Molecular docking supported these findings, indicating that nicofluprole binds at the transmembrane subunit interface rather than within the central pore, consistent with allosteric modulation. These results demonstrate that structural modification of the phenylpyrazole scaffold can redirect target-site engagement from the pore to an allosteric site, explaining the lack of cross-resistance with traditional pore blockers and expanding strategies for GABA receptor-targeted insecticide design.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c09630","type":"journal-article","created":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T07:01:14Z","timestamp":1790838074000},"page":"31519-31526","source":"Crossref","is-referenced-by-count":0,"title":["Structural Modifications\nRedirect Phenylpyrazole Ectoparasiticides\nfrom the Ion Channel Pore to an Allosteric Site on GABA Receptors"],"prefix":"10.1021","volume":"74","author":[{"given":"Tianhao","family":"Zhou","sequence":"first","affiliation":[{"name":"Zhejiang A&F University , , ,","place":["Hangzhou, China, 311300"]},{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zihan","family":"Cheng","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Honglei","family":"Xie","sequence":"additional","affiliation":[{"name":"Ringpu Biology Co. Ltd. , ,","place":["Tianjin, China, 300000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ailing","family":"Liu","sequence":"additional","affiliation":[{"name":"Ringpu Biology Co. Ltd. , ,","place":["Tianjin, China, 300000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shoujun","family":"Li","sequence":"additional","affiliation":[{"name":"Ringpu Biology Co. Ltd. , ,","place":["Tianjin, China, 300000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9590-1319","authenticated-orcid":true,"given":"Xiaomu","family":"Qiao","sequence":"additional","affiliation":[{"name":"Zhejiang A&F University , , ,","place":["Hangzhou, China, 311300"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8336-1562","authenticated-orcid":true,"given":"Jia","family":"Huang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,10,1]]},"reference":[{"issue":"3","key":"2026101408094229400_cit1","doi-asserted-by":"publisher","first-page":"351","DOI":"10.1111\/jphp.12368","article-title":"Ectoparasites\nof Medical and Veterinary Importance: Drug Resistance and the Need\nfor Alternative Control Methods","volume":"67","author":"McNair","year":"2015","journal-title":"J. Pharm. Pharmacol."},{"issue":"6","key":"2026101408094229400_cit2","doi-asserted-by":"publisher","first-page":"595","DOI":"10.1016\/S0965-1748(03)00038-9","article-title":"RdlDv, a Novel GABA-Gated\nChloride Channel Gene from the American Dog Tick Dermacentor Variabilis","volume":"33","author":"Zheng","year":"2003","journal-title":"Insect Biochem. Mol. Biol."},{"key":"2026101408094229400_cit3","doi-asserted-by":"publisher","first-page":"838","DOI":"10.1007\/978-3-662-43978-4_999","article-title":"Ectoparasiticides:\nAntagonists and Modulators of Chloride Channels","volume-title":"Encyclopedia of Parasitology","author":"Turberg","year":"2016"},{"issue":"1","key":"2026101408094229400_cit4","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1002\/ps.3768","article-title":"Differential Mechanisms\nof Action of the Novel \u03b3-Aminobutyric Acid Receptor Antagonist\nEctoparasiticides Fluralaner (A1443) and Fipronil","volume":"71","author":"Asahi","year":"2015","journal-title":"Pest Manag. Sci."},{"issue":"4","key":"2026101408094229400_cit5","doi-asserted-by":"publisher","first-page":"1924","DOI":"10.1002\/ps.7929","article-title":"Functional Validation\nof A2\u2032N Mutation of the RDL GABA Receptor\nagainst Fipronil via Molecular Modeling and Genome Engineering in Drosophila","volume":"80","author":"Qiao","year":"2024","journal-title":"Pest Manag. Sci."},{"key":"2026101408094229400_cit6","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1016\/j.neuro.2016.03.009","article-title":"Mechanisms\nof Resistance to Insecticides Targeting RDL GABA Receptors in Planthoppers","volume":"60","author":"Nakao","year":"2017","journal-title":"Neurotoxicology"},{"issue":"1","key":"2026101408094229400_cit7","doi-asserted-by":"publisher","first-page":"744","DOI":"10.1016\/j.bbrc.2009.11.131","article-title":"The Antiparasitic\nIsoxazoline A1443 Is a Potent Blocker of Insect Ligand-Gated Chloride\nChannels","volume":"391","author":"Ozoe","year":"2010","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"2026101408094229400_cit8","doi-asserted-by":"publisher","DOI":"10.1016\/j.pestbp.2021.105008","article-title":"State-Dependent Inhibition\nof GABA Receptor Channels by the Ectoparasiticide Fluralaner","volume":"181","author":"Kono","year":"2022","journal-title":"Pestic. Biochem. Physiol."},{"key":"2026101408094229400_cit9","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1016\/j.pestbp.2019.11.001","article-title":"Effects of Intersubunit Amino Acid\nSubstitutions on GABA Receptor Sensitivity to the Ectoparasiticide\nFluralaner","volume":"163","author":"Yamato","year":"2020","journal-title":"Pestic. Biochem. Physiol."},{"key":"2026101408094229400_cit10","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioorg.2023.107024","article-title":"Novel Heptafluoroisopropyl\nN-Phenylpyrazole Aryl Amides\nContaining Cyanoalkyl Groups: Design, Synthesis, Insecticidal Activity,\nDocking Studies and Theoretical Calculations","volume":"143","author":"Liu","year":"2024","journal-title":"Bioorg. Chem."},{"issue":"9","key":"2026101408094229400_cit11","doi-asserted-by":"publisher","first-page":"1444","DOI":"10.1038\/s41559-023-02127-4","article-title":"Convergent Resistance to GABA Receptor\nNeurotoxins through Plant\u2013Insect Coevolution","volume":"7","author":"Guo","year":"2023","journal-title":"Nat. Ecol. Evol."},{"issue":"5","key":"2026101408094229400_cit12","doi-asserted-by":"publisher","DOI":"10.3390\/insects15050334","article-title":"Effects of\nRDL GABA Receptor Point Mutants on Susceptibility to Meta-Diamide\nand Isoxazoline Insecticides in Drosophila Melanogaster","volume":"15","author":"Zhou","year":"2024","journal-title":"Insects"},{"issue":"8016","key":"2026101408094229400_cit13","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate Structure\nPrediction of\nBiomolecular Interactions with AlphaFold 3","volume":"630","author":"Abramson","year":"2024","journal-title":"Nature"},{"issue":"6428","key":"2026101408094229400_cit14","doi-asserted-by":"publisher","first-page":"449","DOI":"10.1038\/363449a0","article-title":"A Point\nMutation\nin a Drosophila GABA Receptor Confers Insecticide\nResistance","volume":"363","author":"Ffrench-Constant","year":"1993","journal-title":"Nature"},{"issue":"5","key":"2026101408094229400_cit15","doi-asserted-by":"publisher","first-page":"1957","DOI":"10.1073\/pnas.90.5.1957","article-title":"A Single-Amino Acid Substitution\nin a \u03b3-Aminobutyric\nAcid Subtype a Receptor Locus Is Associated with Cyclodiene Insecticide\nResistance in Drosophila Populations","volume":"90","author":"Ffrench-Constant","year":"1993","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"issue":"4","key":"2026101408094229400_cit16","doi-asserted-by":"publisher","first-page":"366","DOI":"10.1016\/j.ibmb.2013.02.002","article-title":"Meta-Diamide Insecticides\nActing on Distinct Sites of RDL GABA Receptor from Those for Conventional\nNoncompetitive Antagonists","volume":"43","author":"Nakao","year":"2013","journal-title":"Insect Biochem.\nMol. Biol."},{"issue":"1","key":"2026101408094229400_cit17","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1016\/j.ibmb.2013.11.009","article-title":"The Novel\nIsoxazoline Ectoparasiticide Fluralaner: Selective Inhibition of Arthropod\n\u03b3-Aminobutyric Acid- and l-Glutamate-Gated Chloride Channels\nand Insecticidal\/Acaricidal Activity","volume":"45","author":"Gassel","year":"2014","journal-title":"Insect\nBiochem. Mol. Biol."},{"issue":"2","key":"2026101408094229400_cit18","doi-asserted-by":"publisher","first-page":"328","DOI":"10.1111\/bph.12127","article-title":"Advantages\nof an Antagonist: Bicuculline and Other GABA Antagonists","volume":"169","author":"Johnston","year":"2013","journal-title":"Br. J. Pharmacol."},{"issue":"10","key":"2026101408094229400_cit19","doi-asserted-by":"publisher","first-page":"1411","DOI":"10.1016\/j.neuron.2025.12.013","article-title":"Structures\nof the Honeybee\nGABAA RDL Receptor Illuminate Allosteric Modulation","volume":"114","author":"Laboure\u0301","year":"2026","journal-title":"Neuron"},{"issue":"7981","key":"2026101408094229400_cit20","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1038\/s41586-023-06556-w","article-title":"Cryo-EM Structures\nReveal Native\nGABAA Receptor Assemblies and Pharmacology","volume":"622","author":"Sun","year":"2023","journal-title":"Nature"},{"issue":"3","key":"2026101408094229400_cit21","doi-asserted-by":"publisher","first-page":"152","DOI":"10.1584\/jpestics.J15-01","article-title":"Study on the\nStructure and Insecticide Sensitivity of the RDL GABA Receptor","volume":"40","author":"Nakao","year":"2015","journal-title":"J. Pestic. 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Physiol."},{"issue":"6","key":"2026101408094229400_cit24","doi-asserted-by":"publisher","DOI":"10.1371\/JOURNAL.PGEN.1010814","article-title":"G3\u2032MTMD3 in\nthe Insect GABA Receptor Subunit, RDL, Confers\nResistance to Broflanilide and Fluralaner","volume":"19","author":"Zhang","year":"2023","journal-title":"PLoS\nGenet."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c09630","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/40\/31519\/68722268\/acs.jafc.6c09630.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/40\/31519\/68722268\/acs.jafc.6c09630.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T12:29:04Z","timestamp":1791980944000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/40\/31519\/5443135\/Structural-Modifications-Redirect-Phenylpyrazole"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,1]]},"references-count":24,"journal-issue":{"issue":"40","published-online":{"date-parts":[[2026,10,1]]},"published-print":{"date-parts":[[2026,10,14]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c09630","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,1]]}}}