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Cell division cycle\u20102\u2010related kinase 12 (CRK12) has emerged as a genetically essential and chemically validated target in kinetoplastids, yet experimentally supported CRK12\u2010targeting chemotypes against\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>T.\u2009brucei<\/jats:italic>\n                    <\/jats:styled-content>\n                    remain limited, and no experimental CRK12 structure is available. Here, we developed an integrated computational\u2010experimental screening workflow to accelerate CRK12\u2010guided hit discovery, combining ligand\u2010based prescreening, structure\u2010based virtual screening, molecular dynamics (MD) refinement, and experimental validation. An 8.6\u2010million\u2010compound library was prescreened using MACCS fingerprint screening and complex\u2010based pharmacophore screening, followed by staged docking and MD simulations to prioritize candidates for experimental testing. Notably, four of ten purchased compounds showed strong growth inhibition against\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>T.\u2009brucei<\/jats:italic>\n                    <\/jats:styled-content>\n                    at 10\u2009\u03bcM, with IC\n                    <jats:sub>50<\/jats:sub>\n                    values of 6.09, 1.47, 0.81, and 1.33\u2009\u03bcM, demonstrating a promising hit rate for this data\u2010limited target. Binding\u2010mode analysis revealed a conserved hinge\u2010anchoring interaction pattern in the ATP\u2010binding pocket, providing a structural rationale for follow\u2010up analogue design. Overall, this study identifies new chemical starting points for anti\u2010\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>T.\u2009brucei<\/jats:italic>\n                    <\/jats:styled-content>\n                    drug discovery and demonstrates the utility of an AlphaFold2\u2010enabled CRK12\u2010guided screening strategy for targets with limited structural and ligand data.\n                  <\/jats:p>","DOI":"10.1111\/cbdd.70389","type":"journal-article","created":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T03:36:23Z","timestamp":1788147383000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Identification of New Anti\u2010\n                    <i>Trypanosoma brucei<\/i>\n                    Leads: An Integrated\n                    <scp>CRK12<\/scp>\n                    \u2010Guided Virtual Screen With Experimental Validation"],"prefix":"10.1111","volume":"108","author":[{"given":"Qin","family":"Li","sequence":"first","affiliation":[{"name":"Faculty of Applied Sciences Macao Polytechnic University  Macao SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenggong","family":"Fu","sequence":"additional","affiliation":[{"name":"Faculty of Applied Sciences Macao Polytechnic University  Macao SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-5477-2952","authenticated-orcid":false,"given":"Jiayi","family":"Luo","sequence":"additional","affiliation":[{"name":"MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol and Guangdong Provincial Key Laboratory of Aquatic Economic Animals, School of Life Sciences Sun Yat\u2010Sen University  Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4422-4968","authenticated-orcid":false,"given":"Qianqian","family":"Zhang","sequence":"additional","affiliation":[{"name":"Faculty of Applied Sciences Macao Polytechnic University  Macao SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingling","family":"Wang","sequence":"additional","affiliation":[{"name":"Faculty of Applied Sciences Macao Polytechnic University  Macao SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaomeng","family":"Liu","sequence":"additional","affiliation":[{"name":"Faculty of Applied Sciences Macao Polytechnic University  Macao SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-3475-9881","authenticated-orcid":false,"given":"Jiayue","family":"Qiu","sequence":"additional","affiliation":[{"name":"Faculty of Applied Sciences Macao Polytechnic University  Macao SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoqing","family":"Gong","sequence":"additional","affiliation":[{"name":"Faculty of Applied Sciences Macao Polytechnic University  Macao SAR China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2687-741X","authenticated-orcid":false,"given":"Henry H. 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