{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T06:00:48Z","timestamp":1782280848996,"version":"3.54.5"},"posted":{"date-parts":[[2025,11,26]]},"group-title":"Pharmacology and Toxicology","reference-count":28,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,11,26]],"date-time":"2025-11-26T00:00:00Z","timestamp":1764115200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"DOI":"10.13039\/100000049","name":"National Institute on Aging","doi-asserted-by":"crossref","award":["RF1AG084635"],"award-info":[{"award-number":["RF1AG084635"]}],"id":[{"id":"10.13039\/100000049","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,11,26]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Fragment-based drug discovery (FBDD) remains a powerful tool in drug development for targeting a wide range of proteins and identifying new small molecule-based scaffolds. Here, we explored a fragment library of 3,200 compounds using a temperature-related intensity change (TRIC)-based high-throughput screening (HTS) approach, and successfully identified new scaffolds that bind to triggering receptor expressed on myeloid cells 2 (TREM2), a relevant target in neurodegenerative diseases and cancer immunotherapy. We first validated the hits with dose-dependent assays, then chose the three most promising compounds (\n                  <jats:bold>2M06<\/jats:bold>\n                  ,\n                  <jats:bold>6B10<\/jats:bold>\n                  ,\n                  <jats:bold>7G19<\/jats:bold>\n                  ) with binding affinities in the low to medium micromolar range for a \u201cSAR by catalog\u201d study. We screened 29 selected derivatives and subsequently evaluated them with dose-dependent experiments, a thermal shift assay (TSA), selectivity studies with off-targets (LAG-3, TREM1), and finally, in vitro TREM2 activation assays. In this SAR study, derivative\n                  <jats:bold>6B10-9<\/jats:bold>\n                  emerged as the lead compound with moderate TREM2 binding affinity (\n                  <jats:italic>K<\/jats:italic>\n                  <jats:sub>D<\/jats:sub>\n                  = 68.3 \u00b5M) and significant effects on TREM2-dependent phosphorylation of SYK and DAP12 in HEK cells as wells as on microglial phagocytosis in HMC3 cells. Additionally, an in silico analysis revealed that\n                  <jats:bold>6B10-9<\/jats:bold>\n                  forms a stable complex with TREM2 via hydrogen bonding, which maintains its structural integrity during extended molecular dynamics (MD) simulations. These results suggest that\n                  <jats:bold>6B10-9<\/jats:bold>\n                  could serve as a promising lead for future optimizations in the development of small molecule-based TREM2 modulators.\n                <\/jats:p>","DOI":"10.1101\/2025.11.23.690065","type":"posted-content","created":{"date-parts":[[2025,11,26]],"date-time":"2025-11-26T17:50:19Z","timestamp":1764179419000},"source":"Crossref","is-referenced-by-count":0,"title":["High-Throughput Fragment Screening Identifies a New Small Molecule Scaffold that Modulates TREM2 Signaling"],"prefix":"10.64898","author":[{"given":"Natalie","family":"Fuchs","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Farida","family":"El Gaamouch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hossam","family":"Nada","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sungwoo","family":"Cho","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moustafa 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