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PubMed · 37875111

Methotrexate-based PROTACs as DHFR-specific chemical probes.

Sandeep Rana·Patricia Dranchak·Jayme L Dahlin·Laurence Lamy·Wenqing Li·Erin Oliphant·Jonathan H Shrimp·Girish H Rajacharya·Ravi Tharakan·David O Holland·Apryl S Whitten·Kelli M Wilson·Pankaj K Singh·Scott K Durum·Dingyin Tao·Ganesha Rai·James Inglese
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Abstract

Methotrexate (MTX) is a tight-binding dihydrofolate reductase (DHFR) inhibitor, used as both an antineoplastic and immunosuppressant therapeutic. MTX, like folate undergoes folylpolyglutamate synthetase-mediated γ-glutamylation, which affects cellular retention and target specificity. Mechanisms of MTX resistance in cancers include a decrease in MTX poly-γ-glutamylation and an upregulation of DHFR. Here, we report a series of potent MTX-based proteolysis targeting chimeras (PROTACs) to investigate DHFR degradation pharmacology and one-carbon biochemistry. These on-target, cell-active PROTACs show proteasome- and E3 ligase-dependent activity, and selective degradation of DHFR in multiple cancer cell lines. By comparison, treatment with MTX increases cellular DHFR protein expression. Importantly, these PROTACs produced distinct, less-lethal phenotypes compared to MTX. The chemical probe set described here should complement conventional DHFR inhibitors and serve as useful tools for studying one-carbon biochemistry and dissecting complex polypharmacology of MTX and related drugs. Such compounds may also serve as leads for potential autoimmune and antineoplastic therapeutics.

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HumansAntineoplastic AgentsCarbonFolic Acid AntagonistsMethotrexateNeoplasmsProteolysis Targeting ChimeraTetrahydrofolate DehydrogenaseDHFR-HiBiTMTX-Cy5PROTACantifolatesdihydrofolate reductasemethotrexateone-carbon metabolismtargeted protein degradationthymidylate synthaseJournal ArticleResearch Support, N.I.H., IntramuralResearch Support, N.I.H., Extramural

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Sandeep Rana, Patricia Dranchak, Jayme L Dahlin, Laurence Lamy, Wenqing Li, Erin Oliphant, Jonathan H Shrimp, Girish H Rajacharya, Ravi Tharakan, David O Holland, Apryl S Whitten, Kelli M Wilson, Pankaj K Singh, Scott K Durum, Dingyin Tao, Ganesha Rai, James Inglese. 2023-10-23. Methotrexate-based PROTACs as DHFR-specific chemical probes.. https://doi.org/10.1016/j.chembiol.2023.09.014

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