PXD059011-1

PXD059011 is an original dataset announced via ProteomeXchange.

Title	Pyrimethamine and a potent analogue WCDD115 inhibit NRF2 by suppressing DHFR and one-carbon metabolism
Description	Nuclear factor erythroid 2-related factor 2 (NFE2L2/NRF2) is a critical mediator of the cellular oxidative stress response. Aberrant activation of NRF2 is common in many lung and upper aerodigestive cancers, where it promotes tumor progression and confers resistance to chemotherapy, radiation therapy, and immune checkpoint inhibitors. As such, NRF2 therapeutic inhibitors are actively being sought. We previously reported that the antiparasitic drug Pyrimethamine (PYR) inhibits NRF2 in cells and in an NRF2-inducible genetically engineered mouse model. Here we design, synthesize, and define structure-activity relationships across a series of 25 PYR-based derivatives to reveal WCDD115 as a 22-fold more potent inhibitor of NRF2 (57nM versus 1.2mM). PYR is known to inhibit plasmodial and human dihydrofolate reductase (DHFR). We found that WCDD115 inhibits hDHFR with 31-fold greater potency than PYR (144nM versus 4.49mM). Metabolomics showed strong similarities between PYR, WCDD115 and methotrexate. Genetic, pharmacological and metabolic epistasis studies reveal that DHFR inactivation is required for NRF2 suppression by WCDD115 and PYR. Global and targeted proteomics revealed overlapping profiles for WCDD115, PYR and methotrexate, including suppression of NRF2 oxidative stress response and activation of TP53 and the DNA damage response. As such, PYR and its more potent derivative WCDD115 are effective, indirect inhibitors of NRF2 and its antioxidant functions. These data underscore the importance of one-carbon metabolism for the NRF2 signaling pathway and supports a new therapeutic strategy to suppress NRF2-driven cancer biology.
HostingRepository	PRIDE
AnnounceDate	2025-10-13
AnnouncementXML	Submission_2025-10-12_19:29:31.440.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Julius Chembo
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2024-12-18 13:50:06	ID requested
⏵ 1	2025-10-12 19:29:32	announced

10.1016/j.jbc.2025.110659;

Chembo J, Bowman BM, Lapak K, Wilkerson E, Wamsley NT, Paiboonrungruang C, Cho K, Medcalf MR, Wang H, Patti GJ, Dolle RE, Chen X, Zolkind P, Major MB, Pyrimethamine and a potent analog inhibit NRF2 by suppressing one-carbon metabolism. J Biol Chem, 301(10):110659(2025) [pubmed]

submitter keyword: Human, DHFR, Cancer, NRF2

Michel B Major
contact affiliation	Washington University in St Louis
contact email	bmajor@wustl.edu
lab head
Julius Chembo
contact affiliation	Washington University in St. Louis
contact email	juliuschembo@gmail.com
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/10/PXD059011

PRIDE project URI

• PRIDE
  1. PXD059011
     1. Label: PRIDE project
     2. Name: Pyrimethamine and a potent analogue WCDD115 inhibit NRF2 by suppressing DHFR and one-carbon metabolism