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Here, we report our design, synthesis, and evaluation of CDK9 degraders with high selectivity based upon the concept of PROTAC. The representative compound dCDK9-202 demonstrates a DC50 value of 3.5 nM and Dmax &amp;gt; 99% in inducing rapid CDK9 degradation in the TC-71 cell line. Notably, dCDK9-202 achieves significant cell growth inhibition with an IC50 value of 8.5 nM in the TC-71 cell line and also low nanomolar IC50 values in multiple cancer cell lines originating from the lung, liver, bone, and brain. Moreover, intravenous administration of dCDK9-202 effectively inhibits TC-71 tumor growth without any signs of toxicity in mice. This promising CDK9 degrader dCDK9-202 has a high potential for advanced preclinical development in the treatment of CDK9-addicted human cancers.<\/jats:p>","DOI":"10.1021\/acs.jmedchem.5c01111","type":"journal-article","created":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:47:55Z","timestamp":1760032075000},"page":"21172-21186","source":"Crossref","is-referenced-by-count":10,"title":["Discovery of\ndCDK9-202 as a Highly Potent and Selective\nPROTAC CDK9 Degrader with Strong\n                    <i>In Vivo<\/i>\n                    Antitumor\nActivity"],"prefix":"10.1021","volume":"68","author":[{"given":"Lin","family":"Ma","sequence":"first","affiliation":[{"name":"Zhejiang University School of Medicine , , ,","place":["Hangzhou, China, 310029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Long","family":"Xie","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Wang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xian","family":"Guan","sequence":"additional","affiliation":[{"name":"Zhejiang University School of Medicine , , ,","place":["Hangzhou, China, 310029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Zhang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanjun","family":"Guo","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]},{"name":"Children\u2019s Hospital Zhejiang University School of Medicine , , ,","place":["Hangzhou, China, 310052"]},{"name":"National Clinical Research Center for Child Health , , ,","place":["Hangzhou, China, 310052"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiawei","family":"Zhou","sequence":"additional","affiliation":[{"name":"Children\u2019s Hospital Zhejiang University School of Medicine , , ,","place":["Hangzhou, China, 310052"]},{"name":"National Clinical Research Center for Child Health , , ,","place":["Hangzhou, China, 310052"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiyang","family":"Liu","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xingze","family":"Huang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunyu","family":"He","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ye","family":"Chen","sequence":"additional","affiliation":[{"name":"Children\u2019s Hospital Zhejiang University School of Medicine , , ,","place":["Hangzhou, China, 310052"]},{"name":"National Clinical Research Center for Child Health , , ,","place":["Hangzhou, China, 310052"]},{"name":"Cancer Center of Zhejiang University , ,","place":["Hangzhou, China, 310029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liang","family":"Xu","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310058"]},{"name":"Cancer Center of Zhejiang University , ,","place":["Hangzhou, China, 310029"]},{"name":"The Second Affiliated Hospital, Zhejiang University School of Medicine , , ,","place":["Hangzhou, China, 310009"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Han","sequence":"additional","affiliation":[{"name":"Zhejiang University School of Medicine , , ,","place":["Hangzhou, China, 310029"]},{"name":"Cancer Center of Zhejiang University , ,","place":["Hangzhou, China, 310029"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2025,10,9]]},"reference":[{"key":"2026072715524876700_cit1","doi-asserted-by":"publisher","first-page":"862","DOI":"10.17179\/excli2024-7076","article-title":"Cancer therapy by cyclin-dependent\nkinase inhibitors (CDKIs): bench to bedside","volume":"23","author":"Hassanzadeh","year":"2024","journal-title":"EXCLI J."},{"issue":"15","key":"2026072715524876700_cit2","doi-asserted-by":"publisher","first-page":"13147","DOI":"10.1021\/acs.jmedchem.4c01095","article-title":"Genomic\nDiscovery and Structure-Activity\nExploration of a Novel Family of Enzyme-Activated Covalent Cyclin-Dependent\nKinase Inhibitors","volume":"67","author":"Davison","year":"2024","journal-title":"J. Med. Chem."},{"issue":"1","key":"2026072715524876700_cit3","doi-asserted-by":"publisher","first-page":"6316","DOI":"10.1038\/s41467-023-42049-0","article-title":"A cyclin-dependent\nkinase-mediated phosphorylation switch of disordered protein condensation","volume":"14","author":"Valverde","year":"2023","journal-title":"Nat. Commun."},{"issue":"12","key":"2026072715524876700_cit4","doi-asserted-by":"publisher","first-page":"1893","DOI":"10.1080\/10428194.2023.2244102","article-title":"Cyclin-dependent\nkinase-9 in B-cell malignancies: pathogenic role and therapeutic implications","volume":"64","author":"Dominguez","year":"2023","journal-title":"Leuk Lymphoma"},{"key":"2026072715524876700_cit5","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2024.102906","article-title":"The role of intrinsic\nprotein disorder in regulation\nof cyclin-dependent kinases","volume":"88","author":"Phillips","year":"2024","journal-title":"Curr. Opin Struct\nBiol."},{"key":"2026072715524876700_cit6","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbi.2024.110940","article-title":"A systematic review on understanding\nthe mechanistic pathways and clinical aspects of natural CDK inhibitors\non cancer progression.: Unlocking cellular and biochemical mechanisms","volume":"393","author":"Asghar","year":"2024","journal-title":"Chem. Biol. Interact"},{"issue":"10","key":"2026072715524876700_cit7","doi-asserted-by":"publisher","first-page":"1105","DOI":"10.1080\/14737140.2021.1944109","article-title":"The evolution of\ncyclin dependent\nkinase inhibitors in the treatment of cancer","volume":"21","author":"Jhaveri","year":"2021","journal-title":"Expert Rev. 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Cancer\nRes."},{"issue":"23","key":"2026072715524876700_cit13","doi-asserted-by":"publisher","first-page":"15629","DOI":"10.1021\/acs.jmedchem.3c01233","article-title":"Discovery of KB-0742, a Potent, Selective, Orally Bioavailable\nSmall Molecule Inhibitor of CDK9 for MYC-Dependent Cancers","volume":"66","author":"Freeman","year":"2023","journal-title":"J. Med. Chem."},{"issue":"2","key":"2026072715524876700_cit14","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1038\/nchembio.2538","article-title":"Pharmacological perturbation of CDK9\nusing selective\nCDK9 inhibition or degradation","volume":"14","author":"Olson","year":"2018","journal-title":"Nat. Chem. 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Mol. Sci."},{"issue":"4","key":"2026072715524876700_cit18","doi-asserted-by":"publisher","first-page":"1585","DOI":"10.1007\/s00432-022-04135-6","article-title":"Cyclins and cyclin-dependent\nkinases: from biology to tumorigenesis and therapeutic opportunities","volume":"149","author":"Zabihi","year":"2023","journal-title":"J. Cancer Res. Clin Oncol"},{"issue":"5","key":"2026072715524876700_cit19","doi-asserted-by":"publisher","first-page":"823","DOI":"10.1007\/s11684-023-1027-5","article-title":"Discovery of small molecule degraders\nfor modulating cell cycle","volume":"17","author":"Wang","year":"2023","journal-title":"Front Med."},{"issue":"5","key":"2026072715524876700_cit20","doi-asserted-by":"publisher","first-page":"514","DOI":"10.1038\/nchembio.2329","article-title":"Structural basis of\nPROTAC cooperative recognition for selective protein degradation","volume":"13","author":"Gadd","year":"2017","journal-title":"Nat. Chem. 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Biol."},{"issue":"2","key":"2026072715524876700_cit24","doi-asserted-by":"publisher","first-page":"448","DOI":"10.1021\/acs.jmedchem.8b00909","article-title":"Discovery of MD-224\nas a First-in-Class, Highly Potent,\nand Efficacious Proteolysis Targeting Chimera Murine Double Minute\n2 Degrader Capable of Achieving Complete and Durable Tumor Regression","volume":"62","author":"Li","year":"2019","journal-title":"J. Med. Chem."},{"issue":"17","key":"2026072715524876700_cit25","doi-asserted-by":"publisher","first-page":"12831","DOI":"10.1021\/acs.jmedchem.1c00882","article-title":"Strategies\ntoward Discovery of Potent and Orally Bioavailable Proteolysis Targeting\nChimera Degraders of Androgen Receptor for the Treatment of Prostate\nCancer","volume":"64","author":"Han","year":"2021","journal-title":"J. Med. Chem."},{"issue":"18","key":"2026072715524876700_cit26","doi-asserted-by":"publisher","first-page":"13487","DOI":"10.1021\/acs.jmedchem.1c00900","article-title":"Discovery of ARD-2585 as an Exceptionally\nPotent and Orally Active PROTAC Degrader of Androgen Receptor for\nthe Treatment of Advanced Prostate Cancer","volume":"64","author":"Xiang","year":"2021","journal-title":"J.\nMed. Chem."},{"issue":"24","key":"2026072715524876700_cit27","doi-asserted-by":"publisher","first-page":"11218","DOI":"10.1021\/acs.jmedchem.9b01393","article-title":"Discovery of Highly Potent and Efficient\nPROTAC Degraders of Androgen Receptor (AR) by Employing Weak Binding\nAffinity VHL E3 Ligase Ligands","volume":"62","author":"Han","year":"2019","journal-title":"J. Med. Chem."},{"key":"2026072715524876700_cit28","doi-asserted-by":"publisher","first-page":"72","DOI":"10.15212\/amm-2024-0010","article-title":"Application of PROTACs\nin target identification and validation","volume":"3","author":"Liu","year":"2024","journal-title":"Acta\nMater. Med."},{"key":"2026072715524876700_cit29","doi-asserted-by":"publisher","first-page":"104","DOI":"10.1038\/s41580-024-00778-0","article-title":"Mechanisms and regulation\nof substrate degradation by the 26S proteasome","volume":"26","author":"Arkinson","year":"2025","journal-title":"Nat. Rev. Mol. 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Sci."},{"issue":"6","key":"2026072715524876700_cit33","doi-asserted-by":"publisher","DOI":"10.1002\/mco2.575","article-title":"Proteolysis-Targeting\nChimeras (PROTACs)\nin leukemia: overview and future perspectives","volume":"5","author":"Vicente","year":"2024","journal-title":"MedComm"},{"key":"2026072715524876700_cit34","doi-asserted-by":"publisher","DOI":"10.1002\/cmdc.202400267","article-title":"Proteolysis Targeting\nChimeras (PROTACs) in Breast Cancer Therapy","author":"Jin","year":"2024","journal-title":"ChemMedChem."},{"key":"2026072715524876700_cit35","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmech.2024.116613","article-title":"Discovery of bivalent small molecule\ndegraders of cyclin-dependent\nkinase 7 (CDK7)","volume":"276","author":"Ji","year":"2024","journal-title":"Eur. J. Med. Chem."},{"issue":"10","key":"2026072715524876700_cit36","doi-asserted-by":"publisher","first-page":"522","DOI":"10.1016\/j.neo.2020.07.002","article-title":"A highly potent\nPROTAC androgen receptor (AR) degrader ARD-61 effectively inhibits\nAR-positive breast cancer cell growth in vitro tumor growth in vivo","volume":"22","author":"Zhao","year":"2020","journal-title":"Neoplasia"},{"issue":"2","key":"2026072715524876700_cit37","doi-asserted-by":"publisher","first-page":"941","DOI":"10.1021\/acs.jmedchem.8b01631","article-title":"Discovery\nof ARD-69 as a Highly Potent Proteolysis Targeting Chimera (PROTAC)\nDegrader of Androgen Receptor (AR) for the Treatment of Prostate Cancer","volume":"62","author":"Han","year":"2019","journal-title":"J. Med. Chem."},{"issue":"1","key":"2026072715524876700_cit38","doi-asserted-by":"publisher","first-page":"2219470","DOI":"10.1080\/15384047.2023.2219470","article-title":"Targeting CDK9 with selective inhibitors\nor degraders in tumor therapy: an overview of recent developments","volume":"24","author":"Xiao","year":"2023","journal-title":"Cancer Biol. Ther"},{"key":"2026072715524876700_cit39","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmech.2020.113091","article-title":"Discovery\nof selective CDK9 degraders with enhancing antiproliferative activity\nthrough PROTAC conversion","volume":"211","author":"Qiu","year":"2021","journal-title":"Eur. J. Med. Chem."},{"key":"2026072715524876700_cit40","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmech.2023.115774","article-title":"Discovery\nof novel flavonoid-based\nCDK9 degraders for prostate cancer treatment via a PROTAC strategy","volume":"260","author":"Wu","year":"2023","journal-title":"Eur. J. Med. Chem."},{"issue":"16","key":"2026072715524876700_cit41","doi-asserted-by":"publisher","first-page":"11034","DOI":"10.1021\/acs.jmedchem.2c00257","article-title":"Discovery\nof Small-Molecule Degraders of the CDK9-Cyclin T1 Complex for Targeting\nTranscriptional Addiction in Prostate Cancer","volume":"65","author":"Li","year":"2022","journal-title":"J. Med. Chem."},{"issue":"18","key":"2026072715524876700_cit42","doi-asserted-by":"publisher","first-page":"13280","DOI":"10.1021\/acs.jmedchem.3c01264","article-title":"Discovery of ARD-1676\nas a Highly Potent and Orally Efficacious AR\nPROTAC Degrader with a Broad Activity against AR Mutants for the Treatment\nof AR + Human Prostate Cancer","volume":"66","author":"Xiang","year":"2023","journal-title":"J. Med. Chem."},{"issue":"13","key":"2026072715524876700_cit43","doi-asserted-by":"publisher","first-page":"8822","DOI":"10.1021\/acs.jmedchem.3c00405","article-title":"Discovery\nof ARD-2051 as a Potent and Orally Efficacious Proteolysis Targeting\nChimera (PROTAC) Degrader of Androgen Receptor for the Treatment of\nAdvanced Prostate Cancer","volume":"66","author":"Han","year":"2023","journal-title":"J. Med. Chem."},{"key":"2026072715524876700_cit44","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2425812122","article-title":"Discovery\nof an LSD1 PROTAC degrader","volume":"122","author":"Hosseini","year":"2025","journal-title":"Proc. Natl. Acad.\nSci. U. S. A."}],"container-title":["Journal of Medicinal Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jmedchem.5c01111","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jmcmar\/article-pdf\/68\/20\/21172\/66292271\/acs.jmedchem.5c01111.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jmcmar\/article-pdf\/68\/20\/21172\/66292271\/acs.jmedchem.5c01111.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T20:05:48Z","timestamp":1785182748000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jmcmar\/article\/68\/20\/21172\/5234656\/Discovery-of-dCDK9-202-as-a-Highly-Potent-and"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,9]]},"references-count":44,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2025,10,9]]},"published-print":{"date-parts":[[2025,10,23]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jmedchem.5c01111","relation":{},"ISSN":["0022-2623","1520-4804"],"issn-type":[{"value":"0022-2623","type":"print"},{"value":"1520-4804","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,9]]}}}