{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T14:39:23Z","timestamp":1786977563725,"version":"build-2736575974"},"reference-count":57,"publisher":"Wiley","issue":"16","license":[{"start":{"date-parts":[[2025,7,11]],"date-time":"2025-07-11T00:00:00Z","timestamp":1752192000000},"content-version":"am","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,7,11]],"date-time":"2025-07-11T00:00:00Z","timestamp":1752192000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000923","name":"Australian Research Council","doi-asserted-by":"publisher","award":["DP180101781"],"award-info":[{"award-number":["DP180101781"]}],"id":[{"id":"10.13039\/501100000923","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemMedChem"],"published-print":{"date-parts":[[2025,8,16]]},"abstract":"<jats:p>\n                    Pitstop 2, (\n                    <jats:italic>Z<\/jats:italic>\n                    )\u2010\n                    <jats:italic>N<\/jats:italic>\n                    \u2010(5\u2010(4\u2010bromenzylidene)\u20104\u2010oxo\u20104,5\u2010dihydrothiazol\u20102\u2010yl)naphthalene\u20101\u2010sulfonamide (\n                    <jats:bold>1<\/jats:bold>\n                    ) inhibits the clathrin terminal domain\u2013amphiphysin interaction (NTD\u2010PPI) and has been widely used to investigate endocytosis. Herein, the synthesis of 56 novel Pitstop 2 analogues via four discrete focused libraries is reported. Specific modification to the 4\u2010bromonenzylidene moiety is explored, and their ability to inhibit the NTD\u2010PPI interaction is examined by ELISA. In cell endocytosis, effects are measured for selective analogues as is the inhibition of dynamin, another protein involved in the endocytosis process. The most NTD\u2010PPI active analogues retain 2\u2010 and 4\u2010disposed substituents on the aromatic head, with 2,3,4\u2010trihydroxy (\n                    <jats:bold>28<\/jats:bold>\n                    ) the most active (IC\n                    <jats:sub>50<\/jats:sub>\n                    0.94\u2009\u03bc\n                    <jats:sc>m<\/jats:sc>\n                    ). Catechol\u2010free 2,3\u2010dihydroxybenzo[\n                    <jats:italic>b<\/jats:italic>\n                    ][1,4]dioxone (\n                    <jats:bold>54<\/jats:bold>\n                    ) is a more promising lead with an NTD\u2010PPI IC\n                    <jats:sub>50<\/jats:sub>\n                    \u2009=\u20091.16\u2009\u03bc\n                    <jats:sc>m<\/jats:sc>\n                    . The corresponding benzo[\n                    <jats:italic>d<\/jats:italic>\n                    ][1,3]dioxole (\n                    <jats:bold>53<\/jats:bold>\n                    ) is threefold less active suggesting ring size preference at this position. Nine analogues show improved or comparable NTD\u2010PPI activity to Pitstop 2 with IC\n                    <jats:sub>50<\/jats:sub>\n                    values from 0.94 to 2.1\u2009\u03bcM. Heterocyclic analogues are well tolerated and potent inhibitors of CME in U2OS cells, in particular, benzofuran\n                    <jats:bold>67<\/jats:bold>\n                    (NTD\u2010PPI IC\n                    <jats:sub>50<\/jats:sub>\n                    1.5\u2009\u03bc\n                    <jats:sc>m<\/jats:sc>\n                    and CME IC\n                    <jats:sub>50<\/jats:sub>\n                    6.8\u2009\u00b1\u20092.7\u2009\u03bc\n                    <jats:sc>m<\/jats:sc>\n                    ). This positions\n                    <jats:bold>67<\/jats:bold>\n                    as one of the most cell active inhibitors of clathrin\u2010mediated endocytosis yet reported.\n                  <\/jats:p>","DOI":"10.1002\/cmdc.202500321","type":"journal-article","created":{"date-parts":[[2025,6,2]],"date-time":"2025-06-02T22:49:41Z","timestamp":1748904581000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Inhibition of the Clathrin Terminal Domain\u2014Amphiphysin Protein\u2013Protein Interaction. Probing the Pitstop 2 Aromatic Moiety"],"prefix":"10.1002","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0458-5037","authenticated-orcid":false,"given":"Kate","family":"Prichard","sequence":"first","affiliation":[{"name":"Chemistry, School of Environmental &amp; Life Sciences The University of Newcastle  University Drive Callaghan NSW 2308 Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark J.","family":"Robertson","sequence":"additional","affiliation":[{"name":"Chemistry, School of Environmental &amp; Life Sciences The University of Newcastle  University Drive Callaghan NSW 2308 Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5507-0493","authenticated-orcid":false,"given":"Ngoc","family":"Chau","sequence":"additional","affiliation":[{"name":"Cell Signalling Unit Children's Medical Research Institute The University of Sydney  Hawkesbury Road, Westmead Sydney NSW 2145 Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-1062-2881","authenticated-orcid":false,"given":"Jing","family":"Xue","sequence":"additional","affiliation":[{"name":"Cell Signalling Unit Children's Medical Research Institute The University of Sydney  Hawkesbury Road, Westmead Sydney NSW 2145 Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3114-7975","authenticated-orcid":false,"given":"Martin","family":"Neuenschwander","sequence":"additional","affiliation":[{"name":"Marc Nazare \u2010 Chemical Biology Leibniz Institute fur Molecular Pharmacologie  Robert\u2010Roessle\u2010Strasse 10 13125 Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Uwe","family":"Fink","sequence":"additional","affiliation":[{"name":"Marc Nazare \u2010 Chemical Biology Leibniz Institute fur Molecular Pharmacologie  Robert\u2010Roessle\u2010Strasse 10 13125 Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6485-6391","authenticated-orcid":false,"given":"Andre","family":"Horatscheck","sequence":"additional","affiliation":[{"name":"Marc Nazare \u2010 Chemical Biology Leibniz Institute fur Molecular Pharmacologie  Robert\u2010Roessle\u2010Strasse 10 13125 Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1602-2330","authenticated-orcid":false,"given":"Marc","family":"Nazare","sequence":"additional","affiliation":[{"name":"Marc Nazare \u2010 Chemical Biology Leibniz Institute fur Molecular Pharmacologie  Robert\u2010Roessle\u2010Strasse 10 13125 Berlin Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7878-0313","authenticated-orcid":false,"given":"Phillip J.","family":"Robinson","sequence":"additional","affiliation":[{"name":"Cell Signalling Unit Children's Medical Research 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