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Related Experiment Video

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Methotrexate-based PROTACs as DHFR-specific chemical probes.

Sandeep Rana1, Patricia Dranchak1, Jayme L Dahlin1

  • 1Division of Preclinical Innovation, National Center for Advancing Translational Sciences, NIH, Rockville, MD 20850, USA.

Cell Chemical Biology
|October 24, 2023
PubMed
Summary

New proteolysis targeting chimeras (PROTACs) degrade dihydrofolate reductase (DHFR), offering a novel approach to cancer therapy. These DHFR-targeting PROTACs show promise for antineoplastic and autoimmune treatments.

Keywords:
DHFR-HiBiTMTX-Cy5PROTACantifolatesdihydrofolate reductasemethotrexateone-carbon metabolismtargeted protein degradationthymidylate synthase

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Area of Science:

  • Biochemistry
  • Pharmacology
  • Medicinal Chemistry

Background:

  • Methotrexate (MTX) is a dihydrofolate reductase (DHFR) inhibitor used in cancer and autoimmune diseases.
  • MTX efficacy is limited by resistance mechanisms, including reduced poly-glutamylation and DHFR upregulation.
  • MTX poly-glutamylation impacts cellular retention and target specificity.

Purpose of the Study:

  • To develop novel MTX-based proteolysis targeting chimeras (PROTACs) for DHFR degradation.
  • To investigate DHFR degradation pharmacology and its impact on one-carbon biochemistry.
  • To explore PROTACs as potential therapeutic agents for cancer and autoimmune diseases.

Main Methods:

  • Synthesis of MTX-based PROTACs.
  • Assessment of PROTACs' cell-activity, proteasome, and E3 ligase dependency.
  • Evaluation of DHFR degradation in cancer cell lines.
  • Phenotypic analysis comparing PROTACs and MTX treatments.

Main Results:

  • Developed potent, cell-active PROTACs that selectively degrade DHFR in cancer cells.
  • PROTACs exhibit proteasome- and E3 ligase-dependent degradation of DHFR.
  • MTX treatment paradoxically increases cellular DHFR protein expression.
  • PROTACs induce distinct, less toxic phenotypes compared to MTX.

Conclusions:

  • MTX-based PROTACs offer a novel strategy for targeting DHFR, distinct from traditional inhibitors.
  • These PROTACs are valuable chemical probes for studying one-carbon metabolism and MTX polypharmacology.
  • The developed PROTACs show potential as leads for novel antineoplastic and autoimmune therapeutics.