DHFR

Summary

DHFR (dihydrofolate reductase, HGNC:2861) is a protein-coding gene on chromosome 5q14.1, encoding Dihydrofolate reductase (P00374). Catalyzes the reduction of 7,8-dihydrofolate (DHF) to 5,6,7,8-tetrahydrofolate in a NADPH-dependent manner. It is a selective cancer dependency (DepMap: 69.8% of cell lines).

Dihydrofolate reductase converts dihydrofolate into tetrahydrofolate, a methyl group shuttle required for the de novo synthesis of purines, thymidylic acid, and certain amino acids. While the functional dihydrofolate reductase gene has been mapped to chromosome 5, multiple intronless processed pseudogenes or dihydrofolate reductase-like genes have been identified on separate chromosomes. Dihydrofolate reductase deficiency has been linked to megaloblastic anemia. Several transcript variants encoding different isoforms have been found for this gene.

Source: NCBI Gene 1719 — RefSeq curated summary.

At a glance

• Gene–disease (curated): constitutional megaloblastic anemia with severe neurologic disease (Definitive, GenCC)
• GWAS associations: 9
• Clinical variants (ClinVar): 597 total — 26 pathogenic, 11 likely-pathogenic
• Phenotypes (HPO): 25
• Druggable target: yes — 23 molecules with ChEMBL bioactivity
• Cancer dependency (DepMap): dependent in 69.8% of screened cell lines
• MANE Select transcript: NM_000791

Identifiers

Gene identifiers

Field	Value
HGNC ID	HGNC:2861
Approved symbol	DHFR
Name	dihydrofolate reductase
Location	5q14.1
Locus type	gene with protein product
Status	Approved
Aliases	DHFR1
Ensembl gene	ENSG00000228716
Ensembl biotype	protein_coding
OMIM	126060
Entrez	1719

Gene structure

Transcript identifiers

Ensembl transcripts: 8 — 5 protein_coding, 2 retained_intron, 1 protein_coding_CDS_not_defined

ENST00000439211, ENST00000504396, ENST00000505337, ENST00000508282, ENST00000511032, ENST00000513048, ENST00000513314, ENST00000935289

RefSeq mRNA: 3 — MANE Select: NM_000791 NM_000791, NM_001290354, NM_001290357

CCDS: CCDS47240, CCDS78028, CCDS78029

Canonical transcript exons

ENST00000439211 — 6 exons

Exon	Start	End
ENSE00001563315	80626226	80629165
ENSE00001594858	80654008	80654057
ENSE00001752304	80654404	80654983
ENSE00003465725	80633877	80633992
ENSE00003645386	80649389	80649494
ENSE00003668681	80637883	80638009

Expression profiles

Bgee: expression breadth ubiquitous, 290 present calls, max score 98.32.

FANTOM5 (CAGE): breadth ubiquitous, TPM avg 8.0997 / max 78.0778, expressed in 1754 samples.

FANTOM5 promoters (3 alternative TSS)

Promoter ID	TPM avg	Samples expressed
62314	7.8786	1750
62313	0.2084	99
62312	0.0128	5

Top tissues by expression

295 total, by Bgee expression score (0-100, higher = more expressed):

Tissue	Anatomy ID	Expression score	Quality
buccal mucosa cell	CL:0002336	98.32	gold quality
ventricular zone	UBERON:0003053	97.43	gold quality
ganglionic eminence	UBERON:0004023	96.56	gold quality
embryo	UBERON:0000922	96.35	gold quality
trabecular bone tissue	UBERON:0002483	95.28	gold quality
bone marrow	UBERON:0002371	94.37	gold quality
bone marrow cell	CL:0002092	93.45	gold quality
oocyte	CL:0000023	93.03	gold quality
inferior vagus X ganglion	UBERON:0005363	91.65	gold quality
jejunal mucosa	UBERON:0000399	91.54	gold quality
endothelial cell	CL:0000115	91.12	gold quality
rectum	UBERON:0001052	91.09	gold quality
subthalamic nucleus	UBERON:0001906	91.03	gold quality
nephron tubule	UBERON:0001231	90.11	gold quality
medial globus pallidus	UBERON:0002477	90.02	gold quality
tongue squamous epithelium	UBERON:0006919	89.77	gold quality
secondary oocyte	CL:0000655	89.68	gold quality
vermiform appendix	UBERON:0001154	89.68	gold quality
caecum	UBERON:0001153	89.53	gold quality
globus pallidus	UBERON:0001875	89.49	gold quality
duodenum	UBERON:0002114	89.44	gold quality
lymph node	UBERON:0000029	89.43	gold quality
jejunum	UBERON:0002115	89.34	gold quality
spinal cord	UBERON:0002240	89.32	gold quality
thymus	UBERON:0002370	89.27	gold quality
medulla oblongata	UBERON:0001896	89.24	gold quality
liver	UBERON:0002107	89.21	gold quality
C1 segment of cervical spinal cord	UBERON:0006469	89.21	gold quality
renal medulla	UBERON:0000362	89.20	gold quality
colonic epithelium	UBERON:0000397	89.11	gold quality

Single-cell (SCXA)

Detected in 10 experiment(s), a significant marker in 7.

Experiment	Marker?	Max mean expression
E-GEOD-93593	yes	254.25
E-GEOD-76312	yes	236.38
E-GEOD-81383	yes	158.39
E-CURD-112	yes	38.54
E-CURD-122	yes	25.07
E-GEOD-125970	yes	21.22
E-ANND-3	yes	7.63
E-MTAB-6379	no	400.53
E-MTAB-6524	no	182.41
E-MTAB-6108	no	148.74

Regulation

Is transcription factor: no

Upstream regulators (CollecTRI, top): CUX1, E2F1, E2F4, ELK3, MYC, NR0B2, NR3C1, RB1, RBL2, SP1, SP2, TBP, TFDP1

miRNA regulators (miRDB)

101 targeting DHFR, top 30 by miRDB confidence (max_score; target_count = how many genes the miRNA targets in total — lower means more specific):

miRNA	Max score	Avg score	miRNA target_count
HSA-MIR-656-3P	100.00	72.15	2788
HSA-MIR-4795-3P	100.00	74.62	4024
HSA-MIR-4455	100.00	65.48	1587
HSA-MIR-190A-3P	100.00	80.35	5520
HSA-MIR-3163	100.00	77.23	8605
HSA-MIR-4282	99.99	75.36	6408
HSA-MIR-548AJ-3P	99.96	73.38	5345
HSA-MIR-548X-3P	99.96	73.38	5345
HSA-MIR-570-3P	99.96	72.41	4910
HSA-MIR-548J-3P	99.94	72.61	4881
HSA-MIR-335-3P	99.93	73.36	4958
HSA-MIR-548AE-3P	99.93	72.66	4867
HSA-MIR-548AH-3P	99.93	72.54	4872
HSA-MIR-548AM-3P	99.93	72.54	4872
HSA-MIR-548AQ-3P	99.93	72.66	4867
HSA-MIR-10523-5P	99.91	69.22	2038
HSA-MIR-8062	99.88	68.43	995
HSA-MIR-30A-3P	99.87	69.74	2928
HSA-MIR-30D-3P	99.87	69.92	2917
HSA-MIR-30E-3P	99.87	69.68	2942
HSA-MIR-5003-3P	99.85	69.29	2517
HSA-MIR-576-5P	99.84	70.46	2582
HSA-MIR-5010-3P	99.83	70.60	2357
HSA-MIR-4420	99.82	70.08	1624
HSA-MIR-181B-2-3P	99.81	70.06	1646
HSA-MIR-181B-3P	99.81	70.06	1646
HSA-MIR-374C-5P	99.80	72.06	2910
HSA-MIR-655-3P	99.80	72.19	2909
HSA-MIR-2681-5P	99.75	67.64	1655
HSA-MIR-6885-3P	99.75	70.36	3187

Functional genomics

DepMap (CRISPR cell-line fitness): dependent in 69.8% of screened cell lines.

Literature-anchored findings (GeneRIF, showing 40)

• protein folding of dihydrofolate reductases (DHFR) from human, Escherichia coli, and Lactobacillus casei were elucidated and compared using intrinsic Trp fluorescence and fluorescence-detected ANS binding (PMID:11779239)
• studied differences between the regulation of Plasmodium and human dihydrofolate reductases (PMID:11964483)
• Computer modeling studies of the structural role of NADPH binding to active site mutants of human dihydrofolate reductase in complex with piritrexim (PMID:11996001)
• Molecular mechanisms that govern translational regulation of DHFR in response to MTX. Review. (PMID:12084458)
• The crystal structures of two human dihydrofolate reductase (hDHFR) ternary complexes, each with bound NADPH cofactor and a lipophilic antifolate inhibitor, have been determined at atomic resolution. (PMID:12096917)
• Identification of amino acid residues on dihydrofolate reductase protein that mediate binding to its own mRNA (PMID:12384595)
• E2F-responsive dihydrofolate reductase promoter regulates the balance between acetylation and methylation of histone H3 lysine 9 (PMID:12588981)
• RNA polymerase II accumulates in the promoter-proximal region of the dihydrofolate reductase and gamma-actin genes. (PMID:12612070)
• cooperative regulation of dhfr gene transcription by Sp1 and E2F in human osteosarcoma cells, U2OS (PMID:12788094)
• Characterization of a cis-acting regulatory element in dihydrofolate reductase mRNA that functions as a dihydrofolate reductase response element. (PMID:14664697)
• Amplification of the DHFR gene may occur more frequently in the presence of RB1-mediated negative regulation of its activity and can be present at clinical onset in osteosarcoma patients (PMID:14679136)
• The polymorphic alleles that diminish bioavailability of reduced folate in the mother during pregnancy was found to contribute to SB and found to be a 19 bp deletion allele (frequency 0.45) within intron-1 of dihydrofolate reductase (DHFR). (PMID:14735580)
• DHFR exists in two conformations, one bound to DHFR mRNA and the other bound to NADPH (PMID:15817466)
• GroEL bound hDHFR might best be described as a dynamic ensemble of randomly structured conformers (PMID:16116078)
• Point mutations in 5 codons of the DHFR gene, which is known to be related to pyrimethamine resistance, were detected in 15 out of the 50 samples (30%). (PMID:16124424)
• The Expression of baseline dihydrofolate reductase and glycinamide ribonucleotide formyl transferase tended to be higher in responders but this association was not significant. (PMID:16467096)
• switch-like behavior results from a well-studied mechanism for the action of human dihydrofolate reductase (PMID:16735474)
• The 19-bp deletion genotype of DHFR gene was associated with a lower plasma total Homocysteine compared with the wild-type genotype. (PMID:16969375)
• transcriptional repression of the major promoter of the dihydrofolate reductase gene depends on a non-coding transcript initiated from the upstream minor promoter and involves both the direct interaction of the RNA and promoter-specific interference (PMID:17237763)
• We did not find evidence for an effect of the DHFR 19-bp deletion or 9-bp repeat on spina bifida risk in mothers and children. (PMID:17336564)
• DHFR 19-bp deletion polymorphism affects the transcription of DHFR gene in humans (PMID:17413111)
• The DHFR intron 19-bp deletion allele may be a protective neural tube defects genetic factor by increasing DHFR mRNA levels in pregnant women. (PMID:17486595)
• study reports that the 19bp-deletion polymorphism of DHFR acts independently (OR 2.69, 95% CI; 1.00-7.28, p<0.05) and in concert with related folate polymorphisms as a significant risk factor for autism (PMID:17597297)
• The tea polyphenol, epigallocatechin-3-gallate, is an efficient inhibitor of human dihydrofolate reductase. (PMID:17683969)
• Preinvasive cervical lesions display locus specific gene amplification of DHFR, and could be considered as a biological marker for genomic instability in the cervix. (PMID:17917571)
• DHFR27 RNA could repress the luciferase expression in a DHFR-dependent manner when placed upstream of luciferase mRNA. (PMID:18045573)
• analysis of human dihydrofolate reductase folding trajectories inside the GroEL GroES chaperonin cavity and free in solution (PMID:18093916)
• An insertion/deletion polymorphism of the dihydrofolate reductase gene is associated with variation in serum and red blood cell folate concentrations in women (PMID:18247058)
• MDM2 regulates dihydrofolate reductase activity through monoubiquitination. (PMID:18451149)
• our early results showed that neither of MTHFR 677CT and DHFR 19-bp deletion polymorphisms were associated with breast cancer risk (PMID:18498051)
• A 19-base pair deletion polymorphism in dihydrofolate reductase is a functional polymorphism, because it limits assimilation of folic acid into cellular folate stores at high and low folic acid intakes. (PMID:19022952)
• Dhps-437 and dhps-540 are strongly associated with SP treatment failure and should be evaluated further as a method for surveillance of SP-based therapy in DRC. (PMID:19055622)
• the first kinetic parameters for DHFR from pjDHFR and pcDHFR with methotrexate (MTX), trimethoprim (TMP), and its potent analogue, PY957, is reported. (PMID:19196009)
• The nuclear translocation of DHFR was not a pre-requisite for DNA damage induced apoptosis. (PMID:19360472)
• DHFR 19-bp insertion/deletion polymorphism and MTHFR C677T in adult acute lymphoblastic leukaemia (PMID:19536847)
• dihydrofolate reductase knockdown has a role in the anticancer activity of 2-hydroxyoleic acid (PMID:19666584)
• The reduction of folic acid by DHFR per gram of human liver (n = 6) is, on average, less than 2% of that in rat liver at physiological pH. (PMID:19706381)
• DHFR is a novel modulator of beta-catenin and GSK3 signaling (PMID:19727391)
• DNA variants have a role in predisposition to disease-free survival in childhood acute lymphoblastic leukemia (PMID:19861437)
• The 19-base pair deletion polymorphism of DHFR was studied in Japanese. The genotype distribution was wild/wild, 11.9%; wild/deletion, 40.1%; deletion/deletion, 48.0%. Frequencies of wild type and deletion alleles were 0.32 and 0.68, respectively. (PMID:20834190)

Cross-species orthologs

6 orthologs

Organism	Symbol	Gene ID
danio_rerio	dhfr	ENSDARG00000004251
danio_rerio	zgc:153031	ENSDARG00000068876
mus_musculus	Dhfr	ENSMUSG00000021707
rattus_norvegicus		ENSRNOG00000082925
drosophila_melanogaster	Dhfr	FBGN0004087
caenorhabditis_elegans	dhfr-1	WBGENE00007974

Paralogs (2): TYMS (ENSG00000176890), DHFR2 (ENSG00000178700)

Protein

Protein identifiers

Dihydrofolate reductase — P00374 (reviewed: P00374)

All UniProt accessions (3): P00374, B0YJ76, B4DM58

UniProt curated annotations — full annotation on UniProt →

Function. Catalyzes the reduction of 7,8-dihydrofolate (DHF) to 5,6,7,8-tetrahydrofolate in a NADPH-dependent manner. Key enzyme in folate metabolism. Contributes to the nuclear and mitochondrial de novo thymidylate biosynthesis pathway. Catalyzes an essential reaction for de novo glycine and purine synthesis, and for DNA precursor synthesis. Binds its own mRNA and that of DHFR2.

Subunit / interactions. Homodimer. Part of the de novo thymidylate synthesis complex composed at least by SHMT1, DHFR and TYMS. Interacts with SHMT1; this interaction is DNA-dependent and mediates DHFR co-localization with LMNB1.

Subcellular location. Mitochondrion. Cytoplasm. Nucleus.

Tissue specificity. Widely expressed in fetal and adult tissues, including throughout the fetal and adult brains and whole blood. Expression is higher in the adult brain than in the fetal brain.

Post-translational modifications. Sumoylated by UBE2I/UBC9.

Disease relevance. Megaloblastic anemia due to dihydrofolate reductase deficiency (DHFRD) [MIM:613839] An inborn error of metabolism, characterized by megaloblastic anemia and/or pancytopenia, severe cerebral folate deficiency, and cerebral tetrahydrobiopterin deficiency. Clinical features include variable neurologic symptoms, ranging from severe developmental delay and generalized seizures in infancy, to childhood absence epilepsy with learning difficulties, to lack of symptoms. The disease is caused by variants affecting the gene represented in this entry.

Pathway. Cofactor biosynthesis; tetrahydrofolate biosynthesis; 5,6,7,8-tetrahydrofolate from 7,8-dihydrofolate: step 1/1.

Similarity. Belongs to the dihydrofolate reductase family.

Isoforms (2)

UniProt ID	Names	Canonical?
P00374-1	1	yes
P00374-2	2

RefSeq proteins (3): NP_000782, NP_001277283, NP_001277286 (=MANE)

Domains & families (InterPro)

ID	Name	Type
IPR001796	DHFR_dom	Domain
IPR012259	DHFR	Family
IPR017925	DHFR_CS	Conserved_site
IPR024072	DHFR-like_dom_sf	Homologous_superfamily

Pfam: PF00186

Enzyme classification (BRENDA):

• EC 1.5.1.3 — dihydrofolate reductase (BRENDA: 90 organisms, 121 substrates, 914 inhibitors, 438 Km, 293 kcat entries)

Substrate kinetics (BRENDA)

12 substrates with measured Km, best-characterized 12. Km ranges are aggregated across organisms/conditions.

Substrate	Km (mM)	Measurements
7,8-DIHYDROFOLATE	—	241
NADPH	0.0004–0.415	157
DIHYDROFOLATE	0.0006–0.238	8
8-METHYLPTERIN	0.03–0.3	4
FOLATE	0.0041–0.0067	2
NADH	0.268–0.32	2
10-FORMYL-DIHYDROFOLATE	0.003	1
6-HYDROXYMETHYLPTERIN	0.0034	1
DIHYDROPTEROATE	0.0009	1
L-THREO-NEOPTERIN	0.0035	1
NADP+	0.123	1
ACETYLPYRIDINE ADENINE NUCLEOTIDE	—	0

Catalyzed reactions (Rhea), 1 shown:

• (6S)-5,6,7,8-tetrahydrofolate + NADP(+) = 7,8-dihydrofolate + NADPH + H(+) (RHEA:15009)

UniProt features (44 total): strand 15, mutagenesis site 8, binding site 8, helix 5, sequence variant 2, turn 2, chain 1, domain 1, splice variant 1, sequence conflict 1

Structure

Experimental structures (PDB)

89 structures, top 30 by resolution.

PDB	Method	Resolution (Å)
1KMV	X-RAY DIFFRACTION	1.05
1KMS	X-RAY DIFFRACTION	1.09
4M6J	X-RAY DIFFRACTION	1.2
5HSR	X-RAY DIFFRACTION	1.21
3FS6	X-RAY DIFFRACTION	1.23
3GHW	X-RAY DIFFRACTION	1.24
2W3B	X-RAY DIFFRACTION	1.27
3GHC	X-RAY DIFFRACTION	1.3
3GHV	X-RAY DIFFRACTION	1.3
3NTZ	X-RAY DIFFRACTION	1.35
3NU0	X-RAY DIFFRACTION	1.35
3NXO	X-RAY DIFFRACTION	1.35
3NXR	X-RAY DIFFRACTION	1.35
3NXX	X-RAY DIFFRACTION	1.35
4G95	X-RAY DIFFRACTION	1.35
4M6K	X-RAY DIFFRACTION	1.4
2C2S	X-RAY DIFFRACTION	1.4
3F8Y	X-RAY DIFFRACTION	1.45
4KEB	X-RAY DIFFRACTION	1.45
5HQY	X-RAY DIFFRACTION	1.46
5HQZ	X-RAY DIFFRACTION	1.46
5HSU	X-RAY DIFFRACTION	1.46
5HUI	X-RAY DIFFRACTION	1.46
5HVE	X-RAY DIFFRACTION	1.46
2C2T	X-RAY DIFFRACTION	1.5
2W3A	X-RAY DIFFRACTION	1.5
3GI2	X-RAY DIFFRACTION	1.53
3S3V	X-RAY DIFFRACTION	1.53
5SDB	X-RAY DIFFRACTION	1.55
6DAV	X-RAY DIFFRACTION	1.55

Predicted structure (AlphaFold)

Model	pLDDT	Fraction very-high
AF-P00374-F1	96.32	0.97

Functional residue map

Curated UniProt residues grouped by drug-discovery relevance — catalytic, ligand-binding, modification, and mutation-validated positions. Source: UniProtKB sequence features.

Ligand- & substrate-binding residues (8): 10; 16–22; 31–36; 55–57; 65; 71; 77–79; 117–124

Mutagenesis-validated functional residues (8):

Position	Phenotype
23	decreases affinity for nadp and dihydrofolate over 10-fold.
23	strongly decreased affinity for methotrexate. decreases catalytic rate constant 200-fold. decreases affinity for nadp an
32	reduces catalytic rate 5-fold. reduces affinity for dihydrofolate 9-fold; when associated with e-36.
36	reduces catalytic rate 2-fold. reduces affinity for dihydrofolate 9-fold; when associated with r-32.
36	increases affinity for dihydrofolate about 3-fold. reduces affinity for nadph about 3-fold.
36	increases affinity for dihydrofolate about 2-fold. no effect on affinity for nadph.
65	increases affinity for dihydrofolate about 3-fold. no effect on affinity for nadph.
65	increases affinity for dihydrofolate about 15-fold. no effect on affinity for nadph.

Function

Pathways and Gene Ontology

Reactome pathways

3 pathways

ID	Pathway
R-HSA-1474151	Tetrahydrobiopterin (BH4) synthesis, recycling, salvage and regulation
R-HSA-196757	Metabolism of folate and pterines
R-HSA-69205	G1/S-Specific Transcription

MSigDB gene sets: 472 (showing top): BROWNE_HCMV_INFECTION_30MIN_DN, GOBP_RESPONSE_TO_NITROGEN_COMPOUND, MULLIGHAN_NPM1_SIGNATURE_3_UP, HORIUCHI_WTAP_TARGETS_DN, BUYTAERT_PHOTODYNAMIC_THERAPY_STRESS_DN, REACTOME_G1_S_SPECIFIC_TRANSCRIPTION, YAGI_AML_WITH_INV_16_TRANSLOCATION, RODRIGUES_THYROID_CARCINOMA_POORLY_DIFFERENTIATED_UP, GOBP_POLYOL_METABOLIC_PROCESS, GOBP_SUPEROXIDE_METABOLIC_PROCESS, GOBP_REGENERATION, GOBP_NEUROGENESIS, GOBP_RESPONSE_TO_AXON_INJURY, GOBP_CELLULAR_RESPONSE_TO_OXYGEN_CONTAINING_COMPOUND, GOBP_PYRIMIDINE_NUCLEOBASE_METABOLIC_PROCESS

GO Biological Process (10): tetrahydrobiopterin biosynthetic process (GO:0006729), one-carbon metabolic process (GO:0006730), negative regulation of translation (GO:0017148), axon regeneration (GO:0031103), response to methotrexate (GO:0031427), dihydrofolate metabolic process (GO:0046452), tetrahydrofolate metabolic process (GO:0046653), tetrahydrofolate biosynthetic process (GO:0046654), folic acid metabolic process (GO:0046655), regulation of removal of superoxide radicals (GO:2000121)

GO Molecular Function (11): mRNA regulatory element binding translation repressor activity (GO:0000900), mRNA binding (GO:0003729), dihydrofolate reductase activity (GO:0004146), folic acid binding (GO:0005542), NADP binding (GO:0050661), molecular adaptor activity (GO:0060090), NADPH binding (GO:0070402), sequence-specific mRNA binding (GO:1990825), RNA binding (GO:0003723), protein binding (GO:0005515), oxidoreductase activity (GO:0016491)

GO Cellular Component (3): mitochondrion (GO:0005739), cytosol (GO:0005829), cytoplasm (GO:0005737)

Reactome top-level categories

Rollup of top-3 pathways:

Category	Pathways
Metabolism of cofactors	1
Metabolism of water-soluble vitamins and cofactors	1
G1/S Transition	1

GO top-level categories

Rollup of top GO terms by namespace:

Category	Terms
folic acid-containing compound metabolic process	3
dicarboxylic acid metabolic process	2
mRNA binding	2
binding	2
cytoplasm	2
cellular anatomical structure	2
diol biosynthetic process	1
pteridine-containing compound biosynthetic process	1
tetrahydrobiopterin metabolic process	1
small molecule metabolic process	1
translation	1
regulation of translation	1
negative regulation of gene expression	1
negative regulation of protein metabolic process	1
neuron projection regeneration	1
response to axon injury	1
axon development	1
response to nitrogen compound	1
response to oxygen-containing compound	1
folic acid-containing compound biosynthetic process	1
tetrahydrofolate metabolic process	1
removal of superoxide radicals	1
regulation of superoxide metabolic process	1
regulation of cellular response to oxidative stress	1
regulation of response to reactive oxygen species	1
translation repressor activity	1
RNA binding	1
oxidoreductase activity, acting on the CH-NH group of donors, NAD or NADP as acceptor	1
vitamin binding	1
carboxylic acid binding	1
modified amino acid binding	1
heterocyclic compound binding	1
adenyl nucleotide binding	1
molecular_function	1
anion binding	1
NADP binding	1
nucleic acid binding	1
catalytic activity	1
intracellular membrane-bounded organelle	1
intracellular anatomical structure	1

Protein interactions and networks

STRING

3932 interactions, top by confidence (×1000):

Protein A	Protein B	Partner UniProt	Score
DHFR	TYMS	P04818	996
DHFR	DHODH	Q02127	882
DHFR	GART	P22102	876
DHFR	SHMT1	P34896	869
DHFR	ATIC	P31939	863
DHFR	MTHFD1	P11586	862
DHFR	MTHFR	P42898	844
DHFR	TK2	O00142	820
DHFR	SHMT2	P34897	817
DHFR	SPR	P35270	815
DHFR	FPGS	Q05932	810
DHFR	DHPS	P49366	789
DHFR	TCN2	P20062	787
DHFR	QDPR	P09417	774
DHFR	MTR	Q99707	766

IntAct

60 interactions, top by confidence:

A	B	Type	Score
RFXANK	RFXAP	psi-mi:“MI:0914”(association)	0.780
CFTR	ESYT2	psi-mi:“MI:2364”(proximity)	0.710
DHFR	DHFR2	psi-mi:“MI:0915”(physical association)	0.560
SFN	OXSR1	psi-mi:“MI:0914”(association)	0.530
LPAR1	TMEM223	psi-mi:“MI:0914”(association)	0.530
NRAS	ESYT2	psi-mi:“MI:2364”(proximity)	0.480
AIFM1	HAX1	psi-mi:“MI:0914”(association)	0.420
nleF	DHFR	psi-mi:“MI:0915”(physical association)	0.370
FOXB1	DDX39A	psi-mi:“MI:0914”(association)	0.350
FOXL1	DDX39A	psi-mi:“MI:0914”(association)	0.350
HSCB	RBP5	psi-mi:“MI:0914”(association)	0.350
TRAF2	UMAD1	psi-mi:“MI:0914”(association)	0.350
RBCK1	UMAD1	psi-mi:“MI:0914”(association)	0.350
SHARPIN	MAP3K7	psi-mi:“MI:0914”(association)	0.350
TRAF2	TMEM178B	psi-mi:“MI:0914”(association)	0.350
SHARPIN	UMAD1	psi-mi:“MI:0914”(association)	0.350
TNIP2	TMEM178B	psi-mi:“MI:0914”(association)	0.350
LRRK2		psi-mi:“MI:0914”(association)	0.350
ELOVL1	LDHA	psi-mi:“MI:0914”(association)	0.350
SAR1B	UBA6	psi-mi:“MI:0914”(association)	0.350
KCNE3	PIK3R2	psi-mi:“MI:0914”(association)	0.350
HNRNPCL2	SMCHD1	psi-mi:“MI:0914”(association)	0.350
CCT8L2	DVL2	psi-mi:“MI:0914”(association)	0.350
LGI1	APAF1	psi-mi:“MI:0914”(association)	0.350
PIGH	ILVBL	psi-mi:“MI:0914”(association)	0.350
NPAS1	CIBAR1	psi-mi:“MI:0914”(association)	0.350
IL5RA	POTEF	psi-mi:“MI:0914”(association)	0.350

BioGRID (160): GKAP1 (Two-hybrid), DHFR (Synthetic Growth Defect), DHFR (Affinity Capture-MS), DHFR (Affinity Capture-MS), DHFR (Negative Genetic), DHFR (Affinity Capture-MS), CDC73 (Synthetic Lethality), CHEK1 (Synthetic Lethality), WEE1 (Synthetic Lethality), DHFR (Affinity Capture-RNA), DHFR (Affinity Capture-MS), DHFR (Affinity Capture-MS), DHFR (Affinity Capture-MS), DHFR (Affinity Capture-MS), DHFR (Biochemical Activity)

ESM2 similar proteins: A0A1J6KGJ9, A0A314KSQ4, B0BNF8, B4G0F3, B8BKI7, B9N1F9, B9SQI7, C6JS30, E0CSI1, O80526, O82782, P00374, P00375, P00376, P00377, P00378, P04753, P09503, P11172, P27421, P29411, Q05B63, Q28DS0, Q2QNG7, Q2R483, Q5I0K3, Q5NVE1, Q5R421, Q5R514, Q5R8R4, Q5R962, Q5RDZ0, Q5RFI8, Q5ZID6, Q640V9, Q69YN2, Q6YJI5, Q7SYN4, Q7TNK6, Q7Z4G4

Diamond homologs: G4VJD6, O02604, O62583, O81395, P00374, P00375, P00376, P00377, P00378, P04174, P04753, P07382, P07807, P09503, P0A547, P0ABQ4, P0ABQ5, P0ABQ6, P0C0P0, P0C0P1, P11045, P12833, P13922, P13955, P16126, P16184, P17719, P20712, P22573, P22906, P27421, P28019, P31073, P31074, P36591, P43791, P45350, P46103, P51820, P57243

SIGNOR signaling

10 interactions.

A	Effect	B	Mechanism
pemetrexed	down-regulates	DHFR	“chemical inhibition”
E2F1	“up-regulates quantity by expression”	DHFR	“transcriptional regulation”
TFDP1	“up-regulates quantity by expression”	DHFR	“transcriptional regulation”
pralatrexate	“down-regulates activity”	DHFR	“chemical inhibition”
methotrexate	“down-regulates activity”	DHFR	“chemical inhibition”
trimetrexate	“down-regulates activity”	DHFR	“chemical inhibition”
“pemetrexed disodium”	“down-regulates activity”	DHFR	“chemical inhibition”
DHFR	“down-regulates quantity”	dihydrofolate(2-)	“chemical modification”
DHFR	“up-regulates quantity”	(6S)-5,6,7,8-tetrahydrofolate(2-)	“chemical modification”

Enriched among interaction partners

Reactome pathways and GO biological processes over-represented among this gene’s 84 IntAct physical interaction partners (hypergeometric vs the genome-wide background, BH-FDR, gene-set size 15–500, ranked by fold). A functional readout of the neighbourhood — distinct from this gene’s own memberships above, and biased toward well-studied / hub proteins, so read it as themes rather than proof.

Reactome pathways:

Pathway	Partners	Fold	FDR
TNFR1-induced NF-kappa-B signaling pathway	5	31.7×	2e-04

GO biological processes:

GO term	Partners	Fold	FDR
canonical NF-kappaB signal transduction	5	24.4×	1e-03

Disease & clinical

Clinical variants and AI predictions

ClinVar

597 variants total. Per-class counts are floors (≥ shown; pagination cap):

Classification	Count (floor)
Pathogenic	26
Likely pathogenic	11
Uncertain significance	278
Likely benign	198
Benign	18

Top pathogenic / likely-pathogenic (30)

Variant ID	HGVS	Classification
1071775	NM_002439.5(MSH3):c.213_220dup (p.Gln74fs)	Pathogenic
1072664	NM_002439.5(MSH3):c.205_224dup (p.His78fs)	Pathogenic
1074257	NC_000005.9:g.(?79949868)(79952360_?)del	Pathogenic
1366267	NC_000005.9:g.(?79949868)(80064832_?)del	Pathogenic
1755218	NM_002439.5(MSH3):c.122del (p.Thr41fs)	Pathogenic
1771531	NM_002439.5(MSH3):c.139_178del (p.Val47fs)	Pathogenic
2091670	NM_002439.5(MSH3):c.71del (p.Val23_Leu24insTer)	Pathogenic
2424436	NC_000005.9:g.(?79949868)(80074665_?)del	Pathogenic
2424446	NC_000005.9:g.(?79949868)(80171681_?)del	Pathogenic
2451067	NM_002439.5(MSH3):c.205_224del (p.Pro69fs)	Pathogenic
2451077	NM_002439.5(MSH3):c.147_154dup (p.Ala52fs)	Pathogenic
2673497	NM_002439.5(MSH3):c.85C>T (p.Gln29Ter)	Pathogenic
2673506	NM_002439.5(MSH3):c.36del (p.Ala13fs)	Pathogenic
2673531	NM_002439.5(MSH3):c.90dup (p.Thr31fs)	Pathogenic
2673556	NM_002439.5(MSH3):c.85del (p.Gln29fs)	Pathogenic
2673571	NM_002439.5(MSH3):c.61C>T (p.Gln21Ter)	Pathogenic
2807709	NM_002439.5(MSH3):c.153_190del (p.Ala52fs)	Pathogenic
2808231	NM_002439.5(MSH3):c.86del (p.Gln29fs)	Pathogenic
29673	NM_000791.4(DHFR):c.238C>T (p.Leu80Phe)	Pathogenic
29674	NM_000791.4(DHFR):c.458A>T (p.Asp153Val)	Pathogenic
2986392	NM_002439.5(MSH3):c.114del (p.Ser39fs)	Pathogenic
3246482	NC_000005.9:g.(?79950547)(80171681_?)del	Pathogenic
3246487	NC_000005.9:g.(?79949868)(80088673_?)del	Pathogenic
4716925	NM_002439.5(MSH3):c.66_145dup (p.Pro49delinsArgPheTer)	Pathogenic
642642	NC_000005.10:g.(?80654718)(80679103_?)del	Pathogenic
831715	NC_000005.10:g.(?80654049)(80679103_?)del	Pathogenic
1067890	NM_000791.4(DHFR):c.-476C>G	Likely pathogenic
1478214	NM_002439.5(MSH3):c.237_237+1dup	Likely pathogenic
1790284	NM_002439.5(MSH3):c.237+1G>A	Likely pathogenic
2676748	NM_002439.5(MSH3):c.200dup (p.Ala68fs)	Likely pathogenic

SpliceAI

1138 predictions. Top by Δscore:

Variant	Effect	Δscore
5:80633868:TATAC:T	donor_loss	1.0000
5:80633869:ATACT:A	donor_loss	1.0000
5:80633870:TACT:T	donor_loss	1.0000
5:80633871:ACTTA:A	donor_loss	1.0000
5:80633872:CTTAC:C	donor_loss	1.0000
5:80633873:T:TA	donor_loss	1.0000
5:80633874:TACT:T	donor_loss	1.0000
5:80633875:A:AC	donor_gain	1.0000
5:80633876:C:CA	donor_gain	1.0000
5:80633876:CT:C	donor_gain	1.0000
5:80633876:CTCTG:C	donor_gain	1.0000
5:80633992:CCTT:C	acceptor_gain	1.0000
5:80633995:T:TC	acceptor_gain	1.0000
5:80654961:CATA:C	donor_gain	1.0000
5:80654962:ATA:A	donor_gain	1.0000
5:80654963:TA:T	donor_gain	1.0000
5:80654963:TAGTA:T	donor_loss	1.0000
5:80654964:AG:A	donor_loss	1.0000
5:80654965:G:GG	donor_gain	1.0000
5:80654965:GTAG:G	donor_loss	1.0000
5:80654966:T:A	donor_loss	1.0000
5:80629123:AATG:A	donor_gain	0.9900
5:80633875:ACT:A	donor_gain	0.9900
5:80633876:CTC:C	donor_gain	0.9900
5:80633876:CTCT:C	donor_gain	0.9900
5:80633989:CTTC:C	acceptor_gain	0.9900
5:80633992:CC:C	acceptor_loss	0.9900
5:80633993:C:CA	acceptor_loss	0.9900
5:80633993:C:CC	acceptor_gain	0.9900
5:80633994:T:C	acceptor_gain	0.9900

AlphaMissense

1243 scored. Top likely-pathogenic:

Variant	Protein change	am_pathogenicity
5:80649407:A:T	V75D	0.992
5:80654417:A:G	W25R	0.990
5:80654417:A:T	W25R	0.990
5:80649459:A:G	W58R	0.989
5:80649459:A:T	W58R	0.989
5:80649418:T:A	R71S	0.986
5:80649418:T:G	R71S	0.986
5:80654039:G:C	F35L	0.986
5:80654039:G:T	F35L	0.986
5:80654041:A:G	F35L	0.986
5:80649412:A:C	N73K	0.985
5:80649412:A:T	N73K	0.985
5:80649470:C:A	G54V	0.985
5:80637912:A:G	W114R	0.984
5:80637912:A:T	W114R	0.984
5:80654415:C:A	W25C	0.982
5:80654415:C:G	W25C	0.982
5:80629111:A:C	F180L	0.981
5:80629111:A:T	F180L	0.981
5:80629113:A:G	F180L	0.981
5:80649400:G:C	S77R	0.981
5:80649400:G:T	S77R	0.981
5:80649402:T:G	S77R	0.981
5:80649434:C:G	R66P	0.981
5:80649471:C:G	G54R	0.979
5:80649476:A:T	I52N	0.979
5:80637902:C:T	G117D	0.978
5:80637902:C:A	G117V	0.976
5:80654437:C:T	G18D	0.975
5:80654461:G:T	A10D	0.975

dbSNP variants (sampled 300 via entrez): RS1000156862 (5:80652305 G>C), RS1000198608 (5:80648204 T>G), RS1000272435 (5:80634235 A>T), RS1000395606 (5:80630609 G>A,T), RS1000444073 (5:80639287 A>G), RS1000888448 (5:80642264 T>A), RS1000955125 (5:80643398 G>A), RS1000980239 (5:80637992 G>A,C), RS1001316153 (5:80632780 C>G,T), RS1001341936 (5:80650152 C>A,T), RS1001488212 (5:80638464 C>A), RS1001524927 (5:80626617 A>T), RS1001538316 (5:80632248 G>C), RS1001664400 (5:80632361 A>G), RS1001927715 (5:80655940 A>G)

Disease associations

OMIM: gene MIM:126060 | disease phenotypes: MIM:617100, MIM:606764, MIM:613839

GenCC curated gene-disease

Disease	Classification	Inheritance
constitutional megaloblastic anemia with severe neurologic disease	Definitive	Autosomal recessive

Mondo (5): hereditary neoplastic syndrome (MONDO:0015356), endometrial carcinoma (MONDO:0002447), familial adenomatous polyposis 4 (MONDO:0044300), gastrointestinal stromal tumor (MONDO:0011719), constitutional megaloblastic anemia with severe neurologic disease (MONDO:0013456)

Orphanet (4): Inherited cancer-predisposing syndrome (Orphanet:140162), MSH3-related polyposis (Orphanet:480536), Gastrointestinal stromal tumor (Orphanet:44890), Constitutional megaloblastic anemia with severe neurologic disease (Orphanet:319651)

HPO phenotypes

25 total (26 of 25 shown, HPO-id order):

HPO	Term
HP:0000007	Autosomal recessive inheritance
HP:0000952	Jaundice
HP:0000980	Pallor
HP:0001251	Ataxia
HP:0001252	Hypotonia
HP:0001263	Global developmental delay
HP:0001290	Generalized hypotonia
HP:0001321	Cerebellar hypoplasia
HP:0001873	Thrombocytopenia
HP:0001876	Pancytopenia
HP:0001889	Megaloblastic anemia
HP:0002059	Cerebral atrophy
HP:0002121	Generalized non-motor (absence) seizure
HP:0002240	Hepatomegaly
HP:0002421	Poor head control
HP:0003621	Juvenile onset
HP:0005484	Secondary microcephaly
HP:0005518	Increased mean corpuscular volume
HP:0011149	Absence seizure with eyelid myoclonia
HP:0011968	Feeding difficulties
HP:0012446	Decreased CSF 5-methyltetrahydrofolate concentration
HP:0012448	Delayed myelination
HP:0025097	Eyelid myoclonus
HP:0025435	Increased circulating lactate dehydrogenase concentration
HP:0040087	Abnormal blood folate concentration
HP:0012114	Endometrial carcinoma

GWAS associations

9 associations (top):

Study	Trait	p-value
GCST002654_1	Shingles	1.000000e-06
GCST002654_2	Shingles	4.000000e-06
GCST003372_48	Glomerular filtration rate (creatinine)	1.000000e-07
GCST004691_1	Huntington’s disease progression	1.000000e-10
GCST010172_6	Idiopathic downbeat nystagmus	5.000000e-07
GCST010701_1	Cortical surface area (MOSTest)	1.000000e-08
GCST010702_52	Subcortical volume (MOSTest)	8.000000e-10
GCST010703_285	Brain morphology (MOSTest)	4.000000e-15
GCST011618_9	Cortical thickness	1.000000e-15

EFO canonical traits (3, from GWAS)

EFO ID	Trait name
EFO:0008336	disease progression measurement
EFO:0004346	neuroimaging measurement
EFO:0004840	cortical thickness

MeSH disease descriptors (3)

Descriptor	Name	Tree numbers
D046152	Gastrointestinal Stromal Tumors	C04.557.450.565.370; C06.301.371.308; C06.405.249.308
D009386	Neoplastic Syndromes, Hereditary	C04.700; C16.320.700
C565095	Megaloblastic Anemia due to Dihydrofolate Reductase Deficiency (supp.)

Drugs & pharmacology

Drug and pharmacology data

Is drug target: yes

ChEMBL targets (1): CHEMBL202 (SINGLE PROTEIN)

Molecules with ChEMBL bioactivity

23 molecules (phase ≥1), by development phase (incl. off-target/promiscuous compounds). Patent mentions across the top 20 by phase: 998,127 (via chembl_molecule»patent_compound — counts attach to the compound, not the gene–compound relationship, so off-target/promiscuous molecules can dominate).

Molecule	Name	Phase	Patents
CHEMBL119	TRIMETREXATE	4	57,002
CHEMBL1201746	PRALATREXATE	4	14,348
CHEMBL1679	LEUCOVORIN	4	81,374
CHEMBL22	TRIMETHOPRIM	4	56,015
CHEMBL225071	RALTITREXED	4	96,748
CHEMBL225072	PEMETREXED	4	55,761
CHEMBL34259	METHOTREXATE	4	398,396
CHEMBL36	PYRIMETHAMINE	4	19,344
CHEMBL439	SULFADIAZINE	4	31,545
CHEMBL455	SULFACETAMIDE	4	18,129
CHEMBL6067485	GENTAMICIN	4
CHEMBL686	MEFENAMIC ACID	4	61,835
CHEMBL898	DIFLUNISAL	4	54,786
CHEMBL973	TERIFLUNOMIDE	4	7,575
CHEMBL134561	ICLAPRIM	3	962
CHEMBL19633	DIAVERIDINE	2	1,584
CHEMBL280378	EPIROPRIM	2	529
CHEMBL296373	EDATREXATE	2	33,704
CHEMBL376180	AMINOPTERIN	2	346
CHEMBL38434	BREQUINAR	2	8,144
CHEMBL747	CYCLOGUANIL	2
CHEMBL7492	PIRITREXIM	2
CHEMBL4468777	FANOTAPRIM	1

PharmGKB: 1 entry (VIP=true, CPIC=false)

PharmGKB clinical annotations

9 annotations.

Variant	Type	Level	Drugs	Phenotypes
rs1053129	Efficacy	3	methotrexate	Osteosarcoma
rs1105525	Efficacy	3	methotrexate	Acute lymphoblastic leukemia
rs1643650	Efficacy	3	methotrexate	Rheumatoid arthritis
rs1650697	Toxicity	3	pemetrexed	Mesothelioma;Non-Small Cell Lung Carcinoma
rs1650723	Toxicity	3	methotrexate	Osteosarcoma
rs408626	Efficacy,Toxicity	3	methotrexate	Acute lymphoblastic leukemia
rs442767	Toxicity	3	pemetrexed	Non-Small Cell Lung Carcinoma
rs442767	Toxicity	3	methotrexate	Acute lymphoblastic leukemia
rs70991108	Toxicity	3	methotrexate	Acute lymphoblastic leukemia;Drug-induced liver injury;Leukopenia;Thrombocytopenia

PharmGKB variants

15 variants.

Variant	Genes	Level	Score	#Clin annots	Drugs
rs7387	DHFR		0.00	0
rs408626	DHFR, MSH3	3	5.50	1	methotrexate
rs442767	DHFR, MSH3	3	2.25	2	pemetrexed;methotrexate
rs1053129	DHFR	3	2.25	1	methotrexate
rs1105525	DHFR, MSH3	3	2.75	1	methotrexate
rs1643650	DHFR	3	0.00	1	methotrexate
rs1643657	DHFR		0.00	0
rs1650697	DHFR, MSH3	3	2.50	1	pemetrexed
rs1650723	DHFR	3	2.25	1	methotrexate
rs10072026	DHFR, MSH3		0.00	0
rs12517451	DHFR		0.00	0
rs34965641	DHFR		0.00	0
rs70991108	DHFR, MSH3	3	0.50	1	methotrexate
rs3045983	DHFR, MSH3		0.00	0
rs200850015	DHFR, MSH3		0.00	0

GtoPdb / IUPHAR curated pharmacology

(IUPHAR/BPS Guide to Pharmacology — expert-curated)

Target class: enzyme — 1.-.-.- Oxidoreductases

Most potent curated ligand interactions (7 total), top 7:

Ligand	Action	Affinity	Parameter
compound 9 [PMID: 15615544]	Inhibition	10.47	pKi
methotrexate	Inhibition	8.92	pKi
pemetrexed	Inhibition	8.15	pKi
trimetrexate	Inhibition	7.89	pKi
pralatrexate	Inhibition	7.35	pKi
iclaprim	Inhibition	6.11	pKi
diflunisal	Inhibition	4.47	pKi

Binding affinities (BindingDB)

76 measured of 90 human assays (203 total across all organisms); most potent 50 below. Values come from heterogeneous assays and are not directly comparable.

Ligand	Measure	Value	Patent
7-[5,6-dimethyl-2-(1,3-thiazol-2-yl)benzimidazol-1-yl]-6-methylquinazoline-2,4-diamine	KI	2.9 nM	US-8835445: Dihydrofolate reductase inhibitors
7-(2-ethoxynaphthalen-1-yl)-6-methylquinazoline-2,4-diamine	KI	4.5 nM	US-8835445: Dihydrofolate reductase inhibitors
N-[3-[3-(2,4-diaminoquinazolin-7-yl)-4-ethoxyphenyl]phenyl]methanesulfonamide	KI	5.8 nM	US-8835445: Dihydrofolate reductase inhibitors
6-chloro-7-[5,6-dimethyl-2-(1,3-thiazol-2-yl)benzimidazol-1-yl]quinazoline-2,4-diamine	KI	7.6 nM	US-8835445: Dihydrofolate reductase inhibitors
Adamantane-1-carboxylic acid (2,4-diamino-6-methyl-pyrimidin-5-yl)-amide; ethane sulfonate	KI	9 nM
1-[3-(2,4-diamino-6-methylquinazolin-7-yl)phenyl]ethanone	KI	26.3 nM	US-8835445: Dihydrofolate reductase inhibitors
[3-(2,4-diamino-6-methylquinazolin-7-yl)-4-methoxyphenyl]methanol	KI	27.5 nM	US-8835445: Dihydrofolate reductase inhibitors
3-(2,4-diamino-6-methylquinazolin-7-yl)-4-ethoxy-N,N-diethylbenzamide	KI	34 nM	US-8835445: Dihydrofolate reductase inhibitors
4-[2-(2,4-Diamino-5,6,7,8-tetrahydropyrido[3,2-d]pyrimidin-6-yl)ethyl]benzoyl-L-glutamic acid (17a)	IC50	40 nM
4-[2-(2,4-Diamino-5-formyl-5,6,7,8-tetrahydropyrido[3,2-d]pyrimidin-6-yl)ethyl]benzoyl-L-glutamic acid (19a)	IC50	41 nM
7-(3-amino-5-isocyanophenyl)-6-methylquinazoline-2,4-diamine	KI	44 nM	US-8835445: Dihydrofolate reductase inhibitors
7-[3-(isocyanomethyl)phenyl]-6-methylquinazoline-2,4-diamine	KI	44 nM	US-8835445: Dihydrofolate reductase inhibitors
6-methyl-7-(3,4,5-trimethoxyphenyl)quinazoline-2,4-diamine	KI	47.1 nM	US-8835445: Dihydrofolate reductase inhibitors
7-(1H-indol-3-yl)-6-methylquinazoline-2,4-diamine	KI	50.5 nM	US-8835445: Dihydrofolate reductase inhibitors
7-(2-chloro-5-methoxy-4-pyridinyl)-6-methylquinazoline-2,4-diamine	KI	52.4 nM	US-8835445: Dihydrofolate reductase inhibitors
methyl 1-(2,4-diaminoquinazolin-7-yl)-2-(1,3-thiazol-2-yl)benzimidazole-5-carboxylate	KI	53.1 nM	US-8835445: Dihydrofolate reductase inhibitors
[4-[2,4-diamino-6-(imidazol-1-ylmethyl)quinazolin-7-yl]-2-(hydroxymethyl)phenyl]methanol	KI	53.6 nM	US-8835445: Dihydrofolate reductase inhibitors
[5-(2,4-diamino-6-methylquinazolin-7-yl)-4-methoxypyrimidin-2-yl]methanol	KI	54.7 nM	US-8835445: Dihydrofolate reductase inhibitors
7-(3-isocyanophenyl)-6-methylquinazoline-2,4-diamine	KI	55 nM	US-8835445: Dihydrofolate reductase inhibitors
[5-chloro-4-(2,4-diamino-6-methylquinazolin-7-yl)-2-pyridinyl]methanol	KI	56 nM	US-8835445: Dihydrofolate reductase inhibitors
4-[2-(2-Diamino-4-hydroxy-5,6,7,8-tetrahydropyrido[3,2-d]pyrimidin-6-yl)ethyl]benzoyl-L-glutamic acid (18a)	IC50	59 nM
6-ethyl-5-[3-(3-methoxy-5-pyridin-4-ylphenyl)prop-1-ynyl]pyrimidine-2,4-diamine	IC50	61 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
7-[5,6-dimethyl-2-(1,3-thiazol-2-yl)benzimidazol-1-yl]quinazoline-2,4-diamine	KI	93.5 nM	US-8835445: Dihydrofolate reductase inhibitors
CHEMBL3234115	IC50	97 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
6-ethyl-5-[3-(2-methoxy-5-phenylphenyl)but-1-ynyl]pyrimidine-2,4-diamine	IC50	100 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
7-(4-aminophenyl)-6-methylquinazoline-2,4-diamine	KI	138 nM	US-8835445: Dihydrofolate reductase inhibitors
6-ethyl-5-(3-(5-methoxybiphenyl-3-yl)prop-1-ynyl)pyrimidine-2,4-diamine	IC50	140 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
6-ethyl-5-[3-(3-pyrimidin-5-ylphenyl)prop-1-ynyl]pyrimidine-2,4-diamine	IC50	160 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
[1-(2,4-diamino-6-methylquinazolin-7-yl)-6-methoxy-2-(1,3-thiazol-2-yl)benzimidazol-5-yl]methanol	KI	170 nM	US-8835445: Dihydrofolate reductase inhibitors
6-methyl-7-(1H-pyrrolo[2,3-b]pyridin-5-yl)quinazoline-2,4-diamine	KI	235 nM	US-8835445: Dihydrofolate reductase inhibitors
7-(1H-indol-4-yl)-6-methylquinazoline-2,4-diamine	KI	245 nM	US-8835445: Dihydrofolate reductase inhibitors
3-[2-(2,4-diaminoquinazolin-7-yl)-5,6-dimethylbenzimidazol-1-yl]propan-1-ol	KI	263 nM	US-8835445: Dihydrofolate reductase inhibitors
6-ethyl-5-[3-(3-pyridin-3-ylphenyl)prop-1-ynyl]pyrimidine-2,4-diamine	IC50	290 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
5-(3-(biphenyl-3-yl)prop-1-ynyl)-6-ethylpyrimidine-2,4-diamine	IC50	290 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
4-[2-(2-Amino-4-hydroxy-5-allyl-5,6,7,8-tetrahydropyrido[3,2-d]pyrimidin-6-yl)ethyl]benzoyl-L-glutamic acid (23a)	IC50	298 nM
6-[(benzyloxy)methyl]-5-(4-{[(4-chlorophenyl)methyl]amino}phenyl)pyrimidine-2,4-diamine	IC50	300 nM
6-ethyl-5-(3-(4-methoxybiphenyl-3-yl)prop-1-ynyl)pyrimidine-2,4-diamine	IC50	300 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
7-(5-methyl-2-thiophen-2-ylbenzimidazol-1-yl)quinazoline-2,4-diamine	KI	314 nM	US-8835445: Dihydrofolate reductase inhibitors
[1-(2,4-diamino-6-chloroquinazolin-7-yl)-6-methoxy-2-(1,3-thiazol-2-yl)benzimidazol-5-yl]methanol	KI	320 nM	US-8835445: Dihydrofolate reductase inhibitors
1-[3-(2,4-diaminoquinazolin-7-yl)phenyl]ethanone	KI	323 nM	US-8835445: Dihydrofolate reductase inhibitors
6-ethyl-5-[3-(2-methoxy-5-pyridin-4-ylphenyl)prop-1-ynyl]pyrimidine-2,4-diamine	IC50	330 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
7-(2,5-dimethoxyphenyl)quinazoline-2,4-diamine	KI	345 nM	US-8835445: Dihydrofolate reductase inhibitors
CHEMBL2335416	IC50	400 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
6-ethyl-5-[3-(3-pyridin-4-ylphenyl)prop-1-ynyl]pyrimidine-2,4-diamine	IC50	520 nM	US-8853228: Heterocyclic analogs of propargyl-linked inhibitors of dihydrofolate reductase
7-(5,6-dimethyl-2-pyridin-2-ylbenzimidazol-1-yl)quinazoline-2,4-diamine	KI	572 nM	US-8835445: Dihydrofolate reductase inhibitors
7-[2-(3,3-difluorobutylsulfanyl)-6-methoxybenzimidazol-1-yl]quinazoline-2,4-diamine	KI	586 nM	US-8835445: Dihydrofolate reductase inhibitors
7-(5,6-dimethyl-2-thiophen-3-ylbenzimidazol-1-yl)quinazoline-2,4-diamine	KI	663 nM	US-8835445: Dihydrofolate reductase inhibitors
4-[2-[2,4-Diamino-5-(2,3-dibromopropyl)-5,6,7,8-tetrahydropyrido[3,2-d]pyrimidin-6-yl]ethyl]benzoyl-L-glutamic acid (21a)	IC50	680 nM
7-(4-aminophenyl)quinazoline-2,4-diamine	KI	702 nM	US-8835445: Dihydrofolate reductase inhibitors
4-[2-(2,4-Diamino-5-allyl-5,6,7,8-tetrahydropyrido[3,2-d]pyrimidin-6-yl)ethyl]benzoyl-L-glutamic acid (20a)	IC50	706 nM

ChEMBL bioactivities

1537 potent at pChembl≥5 of 1792 total, top 50 by pChembl (potency: 10 = 0.1 nM, 6 = 1 µM).

pChembl	Type	Value	Unit	Molecule
11.00	Ki	0.01	nM	METHOTREXATE
11.00	Ki	0.01	nM	CHEMBL305177
10.87	Ki	0.0134	nM	PRALATREXATE
10.60	Ki	0.025	nM	PIRITREXIM
10.47	Ki	0.034	nM	CHEMBL390990
10.42	Ki	0.038	nM	METHOTREXATE
10.40	Ki	0.04	nM	TRIMETREXATE
10.11	Ki	0.077	nM	CHEMBL388501
10.11	Ki	0.077	nM	METHOTREXATE
10.00	Ki	0.1	nM	CHEMBL369258
10.00	Ki	0.1	nM	CHEMBL171765
10.00	Ki	0.1	nM	CHEMBL170760
10.00	Kd	0.1	nM	CHEMBL135937
9.97	Ki	0.107	nM	CHEMBL35364
9.96	Ki	0.11	nM	METHOTREXATE
9.92	Ki	0.12	nM	CHEMBL443763
9.88	Ki	0.131	nM	CHEMBL284943
9.75	Ki	0.179	nM	METHOTREXATE
9.74	Ki	0.183	nM	CHEMBL35427
9.70	Ki	0.2	nM	CHEMBL170837
9.70	Ki	0.2	nM	CHEMBL352960
9.70	Kd	0.2	nM	CHEMBL135937
9.68	Ki	0.21	nM	AMINOPTERIN
9.54	Ki	0.29	nM	CHEMBL354249
9.52	Ki	0.3	nM	CHEMBL170758
9.41	IC50	0.39	nM	CHEMBL251427
9.40	Ki	0.4	nM	CHEMBL424170
9.40	Ki	0.4	nM	CHEMBL171494
9.38	IC50	0.42	nM	TRIMETREXATE
9.30	Kd	0.5	nM	CHEMBL135581
9.27	Ki	0.54	nM	CHEMBL162476
9.22	IC50	0.6	nM	METHOTREXATE
9.22	Kd	0.6	nM	CHEMBL135581
9.16	Ki	0.69	nM	CHEMBL168069
9.15	IC50	0.7	nM	METHOTREXATE
9.14	IC50	0.72	nM	METHOTREXATE
9.10	Ki	0.8	nM	CHEMBL301030
9.09	IC50	0.82	nM	METHOTREXATE
9.08	IC50	0.84	nM	CHEMBL349572
9.00	IC50	1	nM	METHOTREXATE
9.00	IC50	1	nM	CHEMBL36083
8.96	IC50	1.1	nM	CHEMBL18155
8.93	IC50	1.18	nM	METHOTREXATE
8.92	IC50	1.2	nM	METHOTREXATE
8.89	IC50	1.3	nM	CHEMBL2261432
8.89	IC50	1.3	nM	CHEMBL83644
8.87	IC50	1.35	nM	CHEMBL160505
8.87	IC50	1.34	nM	CHEMBL160699
8.85	IC50	1.4	nM	TRIMETREXATE
8.85	IC50	1.4	nM	METHOTREXATE

PubChem BioAssay actives

1453 with measured affinity, of 3493 total; 50 most potent distinct compounds. Largely complementary to BindingDB; screening values are coarse (µM, 4 dp), so sub-nM hits tie at the floor.

Compound	Assay	Type	Value	Unit
2-[[4-carboxy-4-[[4-[(2,4-diaminoquinazolin-6-yl)methylamino]benzoyl]amino]butyl]carbamoyl]benzoic acid	57132: Compound was evaluated for the inhibition of Dihydrofolate reductase at concentration ranged from 0.15-0.50 uM	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[(2,4-diaminopyrido[2,3-d]pyrimidin-6-yl)methylamino]benzoyl]amino]butyl]carbamoyl]benzoic acid	57132: Compound was evaluated for the inhibition of Dihydrofolate reductase at concentration ranged from 0.15-0.50 uM	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[(2,4-diamino-5-chloroquinazolin-6-yl)methylamino]benzoyl]amino]butyl]carbamoyl]benzoic acid	57132: Compound was evaluated for the inhibition of Dihydrofolate reductase at concentration ranged from 0.15-0.50 uM	ki	<0.0001	uM
(2S)-2-[6-[(2,4-diaminopteridin-6-yl)methylamino]-3-oxo-1H-isoindol-2-yl]pentanedioic acid	282637: Inhibition of human DHFR	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[(2,4-diamino-5-methylquinazolin-6-yl)methylamino]benzoyl]amino]butyl]carbamoyl]benzoic acid	57132: Compound was evaluated for the inhibition of Dihydrofolate reductase at concentration ranged from 0.15-0.50 uM	ki	<0.0001	uM
Methotrexate	1314225: Inhibition of recombinant human DHFR assessed as reduction in NADPH oxidation using dihydrofolate as substrate by fluorescence spectrophotometric analysis	ki	<0.0001	uM
Pralatrexate	1894187: Inhibition of DHFR (unknown origin) at pH 6.7	ki	<0.0001	uM
5-(3-chlorophenyl)-6-[2-[2-(3-phenylpropoxy)phenyl]ethyl]pyrimidine-2,4-diamine;hydrochloride	1611287: Inhibition of human dihydrofolate reductase using dihydrofolate as substrate in presence of NADPH by UV-vis spectrophotometry analysis	ki	<0.0001	uM
(2S)-2-[[4-[(2,4-diaminopteridin-6-yl)methylamino]benzoyl]amino]pentanedioic acid	219254: Inhibitory activity against Wild-type human DHFR	ki	<0.0001	uM
8-methyl-7-pentan-3-ylpyrrolo[3,2-f]quinazoline-1,3-diamine	1260819: Competitive inhibition of human recombinant DHFR preincubated for 2 mins followed by substrate addition in presence of dihydrofolate	ki	<0.0001	uM
2-[[4-[(2,4-diaminoquinazolin-6-yl)methylamino]benzoyl]amino]pentanedioic acid	219254: Inhibitory activity against Wild-type human DHFR	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[(2,4-diaminopteridin-6-yl)methylamino]benzoyl]amino]butyl]carbamoyl]benzoic acid	57132: Compound was evaluated for the inhibition of Dihydrofolate reductase at concentration ranged from 0.15-0.50 uM	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[(2,4-diamino-5-methylpyrido[2,3-d]pyrimidin-6-yl)methylamino]benzoyl]amino]butyl]carbamoyl]benzoic acid	57132: Compound was evaluated for the inhibition of Dihydrofolate reductase at concentration ranged from 0.15-0.50 uM	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[(6,8-diaminoquinoxalin-2-yl)methylamino]benzoyl]amino]butyl]carbamoyl]benzoic acid	57138: Inhibitory activity against human recombinant Dihydrofolate reductase	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[1-(6,8-diaminoquinoxalin-2-yl)pent-4-yn-2-yl]benzoyl]amino]butyl]carbamoyl]benzoic acid	57138: Inhibitory activity against human recombinant Dihydrofolate reductase	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[2-(6,8-diaminoquinoxalin-2-yl)ethyl]benzoyl]amino]butyl]carbamoyl]benzoic acid	57138: Inhibitory activity against human recombinant Dihydrofolate reductase	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[1-(6,8-diaminoquinoxalin-2-yl)butan-2-yl]benzoyl]amino]butyl]carbamoyl]benzoic acid	57138: Inhibitory activity against human recombinant Dihydrofolate reductase	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[2-(6,8-diaminonaphthalen-2-yl)ethyl]benzoyl]amino]butyl]carbamoyl]benzoic acid	57138: Inhibitory activity against human recombinant Dihydrofolate reductase	ki	<0.0001	uM
2-[[4-carboxy-4-[[4-[(2,4-diaminopyrido[3,2-d]pyrimidin-6-yl)methylamino]benzoyl]amino]butyl]carbamoyl]benzoic acid	57132: Compound was evaluated for the inhibition of Dihydrofolate reductase at concentration ranged from 0.15-0.50 uM	ki	<0.0001	uM
2-[[4-[(2,4-diaminopyrido[2,3-d]pyrimidin-6-yl)methylamino]naphthalene-1-carbonyl]amino]pentanedioic acid	56168: Tested for inhibition against Dihydrofolate reductase from Pneumocystis carinii	ic50	<0.0001	uM
6-[(2,5-dimethoxyphenyl)methyl]-5-methylpyrido[2,3-d]pyrimidine-2,4-diamine	57133: Inhibition of human recombinant Dihydrofolate reductase enzyme	ki	<0.0001	uM
Trimethoprim	1179551: Binding affinity to DHFR (unknown origin) by NMR analysis	ki	<0.0001	uM
5-methyl-6-[(3,4,5-trimethoxyanilino)methyl]quinazoline-2,4-diamine	1915711: Inhibition of DHFR (unknown origin) assessed as inhibition constant	ki	<0.0001	uM
2-[[4-[3-carboxypropyl-[(2,4-diaminopteridin-6-yl)methyl]amino]benzoyl]amino]pentanedioic acid	56458: Inhibitory activity against Trypanosoma cruzi dihydrofolate reductase	ki	0.0001	uM
2-[[4-[4-carboxybutyl-[(2,4-diaminopteridin-6-yl)methyl]amino]benzoyl]amino]pentanedioic acid	56458: Inhibitory activity against Trypanosoma cruzi dihydrofolate reductase	ki	0.0001	uM
2-amino-5-methyl-8-(2-methylpropyl)-2,3-dihydropyrido[2,3-d]pyrimidin-4-one	57126: Tested for the apparent dissociation constant for binding of compound to ternary NADPH complex with human dihydrofolate reductase using equation 4	kd	0.0001	uM
2-[6-[(2,4-diaminopteridin-6-yl)methylamino]-3-oxo-1H-isoindol-2-yl]pentanedioic acid	282637: Inhibition of human DHFR	ki	0.0001	uM
7-tert-butyl-8-ethylpyrrolo[3,2-f]quinazoline-1,3-diamine	57133: Inhibition of human recombinant Dihydrofolate reductase enzyme	ki	0.0001	uM
7-butan-2-yl-8-ethylpyrrolo[3,2-f]quinazoline-1,3-diamine	57133: Inhibition of human recombinant Dihydrofolate reductase enzyme	ki	0.0001	uM
7-pentan-3-yl-8-propan-2-ylpyrrolo[3,2-f]quinazoline-1,3-diamine	57133: Inhibition of human recombinant Dihydrofolate reductase enzyme	ki	0.0001	uM
2-[[4-[5-carboxypentyl-[(2,4-diaminopteridin-6-yl)methyl]amino]benzoyl]amino]pentanedioic acid	56458: Inhibitory activity against Trypanosoma cruzi dihydrofolate reductase	ki	0.0002	uM
8-tert-butyl-7-propan-2-ylpyrrolo[3,2-f]quinazoline-1,3-diamine	57133: Inhibition of human recombinant Dihydrofolate reductase enzyme	ki	0.0002	uM
8-ethyl-7-pentan-3-ylpyrrolo[3,2-f]quinazoline-1,3-diamine	57133: Inhibition of human recombinant Dihydrofolate reductase enzyme	ki	0.0002	uM
(2R)-2-[[4-[2-(2,4-diamino-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-6-yl)ethyl]benzoyl]amino]-2-methylpentanedioic acid	57139: Inhibitory activity against recombinant human dihydrofolate reductase (DHFR)	ki	0.0003	uM
5-[(2-fluorophenyl)methoxy]quinazoline-2,4-diamine	313040: Inhibition of human recombinant DHFR	ic50	0.0004	uM
8-methyl-7-(2-methylbutan-2-yl)pyrrolo[3,2-f]quinazoline-1,3-diamine	57133: Inhibition of human recombinant Dihydrofolate reductase enzyme	ki	0.0004	uM
7-pentan-3-yl-8-propylpyrrolo[3,2-f]quinazoline-1,3-diamine	57133: Inhibition of human recombinant Dihydrofolate reductase enzyme	ki	0.0004	uM
2-amino-7-methyl-8-propyl-2,3-dihydropyrido[2,3-d]pyrimidin-4-one	57123: Tested for the apparent dissociation constant for binding of compound to binary NADPH complex with human dihydrofolate reductase using equation 4	kd	0.0005	uM
2-[[4-[(2,4-diaminoquinazolin-5-yl)methylamino]benzoyl]amino]pentanedioic acid	219254: Inhibitory activity against Wild-type human DHFR	ki	0.0005	uM
(2R)-2-[[5-[2-(2,4-diamino-5,6,7,8-tetrahydropyrido[2,3-d]pyrimidin-6-yl)ethyl]thiophene-2-carbonyl]amino]-2-methylpentanedioic acid	57139: Inhibitory activity against recombinant human dihydrofolate reductase (DHFR)	ki	0.0007	uM
5-[[4-methoxy-3,5-bis[(E)-prop-1-enyl]phenyl]methyl]pyrimidine-2,4-diamine	56001: Antibacterial activity against Escherichia coli	ki	0.0008	uM
(2R,4R)-2-[[4-[(2,4-diaminopteridin-6-yl)methyl-methylamino]benzoyl]amino]-4-fluoropentanedioic acid	56966: Inhibitory activity against Dihydrofolate reductase (DHFR) isolated from CCRF-CEM human leukemia cells in experiment 1	ic50	0.0008	uM
4H-pyrrolo[3,2-f]quinazoline-1,3-diamine	1260819: Competitive inhibition of human recombinant DHFR preincubated for 2 mins followed by substrate addition in presence of dihydrofolate	ki	0.0010	uM
5-amino-2-[[4-[(2,4-diaminopteridin-6-yl)methyl-methylamino]benzoyl]amino]pentanoic acid	56974: Inhibitory concentration against dihydrofolate reductase enzyme (DHFR) enzyme isolated from CCRF-CEM human leukemia cells. value mentioned is from literature	ic50	0.0010	uM
5-amino-2-[[4-[(2,4-diaminopteridin-6-yl)methyl-methylamino]benzoyl]amino]-4,4-difluoropentanoic acid	56973: Inhibitory concentration against dihydrofolate reductase enzyme (DHFR) enzyme isolated from CCRF-CEM human leukemia cells.	ic50	0.0013	uM
2-[[4-[(2,4-diaminopteridin-6-yl)methyl-methylamino]benzoyl]amino]-3,3-difluoropentanedioic acid	56967: Inhibitory activity against Dihydrofolate reductase (DHFR) isolated from CCRF-CEM human leukemia cells in experiment 2	ic50	0.0013	uM
2-[[4-[(2,4-diaminopteridin-6-yl)methyl-methylamino]benzoyl]amino]-4-fluoropentanedioic acid	56966: Inhibitory activity against Dihydrofolate reductase (DHFR) isolated from CCRF-CEM human leukemia cells in experiment 1	ic50	0.0014	uM
6-[(3,5-dimethoxyphenyl)methyl]-5-methylpyrido[2,3-d]pyrimidine-2,4-diamine	56975: Inhibitory concentration against dihydrofolate reductase of rat liver	ic50	0.0015	uM
5-(3-chlorophenyl)-6-ethylpyrimidine-2,4-diamine	1533249: Inhibition of human DHFR	ki	0.0016	uM
6-fluoro-2-[4-(2-fluorophenyl)phenyl]-3-methylquinoline-4-carboxylic acid	1439522: Inhibition of recombinant human N-terminal truncated GST-tagged DHFR (31 to 395 residues) expressed in Escherichia coli BL21(DE3) pyrD using DHO as substrate preincubated for 5 mins followed by substrate addition measured for 5 mins by DCIP oxidation based CoQ10 enzyme coupled assay	ic50	0.0018	uM

CTD chemical–gene interactions

114 total (human), top 30 by PubMed support.

Chemical	Actions (top 5)	PubMed papers
Methotrexate	affects cotreatment, affects binding, decreases activity, increases response to substance, decreases response to substance (+5 more)	13
Folic Acid	increases expression, decreases reaction, decreases abundance, affects expression, affects response to substance (+2 more)	6
bisphenol A	decreases expression, affects expression, affects cotreatment, decreases methylation	4
sodium arsenite	decreases expression, increases expression, increases mutagenesis	4
Tobacco Smoke Pollution	decreases expression, increases methylation	4
Benzo(a)pyrene	increases expression, decreases expression	3
Fluorouracil	affects expression, increases expression	3
perfluorooctanoic acid	decreases expression	2
epigallocatechin gallate	affects binding, decreases activity, affects cotreatment, increases expression	2
Air Pollutants	increases expression, decreases expression, increases abundance	2
Arsenic	affects methylation, affects response to substance	2
Cisplatin	decreases expression, increases reaction, increases expression	2
Leucovorin	increases expression, decreases abundance, decreases reaction	2
7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide	increases expression, decreases expression	2
aristolochic acid I	increases expression	1
GSK-J4	decreases expression	1
afuresertib	decreases expression	1
TAK-243	increases sumoylation	1
syringic acid	decreases activity	1
methylmercuric chloride	decreases expression	1
ferulic acid	decreases activity	1
naringin	decreases activity	1
triphenyl phosphate	affects expression	1
propionaldehyde	decreases expression	1
methylselenic acid	decreases expression	1
sodium arsenate	decreases expression	1
methylparaben	decreases expression	1
tris(1,3-dichloro-2-propyl)phosphate	decreases expression	1
cobaltous chloride	decreases expression	1
alpha-cobratoxin	decreases expression, decreases reaction	1

ChEMBL screening assays

457 unique, capped per target: 426 binding, 16 admet, 12 functional, 3 toxicity

Representative assays (with source publication via chembl_document):

Assay ID	Type	Description	Source paper
CHEMBL1017358	Binding	Inhibition of DHFR	A new bastadin from the sponge Psammaplysilla purpurea. — J Nat Prod
CHEMBL3867373	ADMET	Inhibition of recombinant human DHFR expressed in Escherichia coli preincubated for 15 mins followed by addition of DHF as substrate and NADPH measured after 60 mins by resazurin/diaphorase coupled assay	Discovery of Potent and Selective Leads against Toxoplasma gondii Dihydrofolate Reductase via Structure-Based Design. — ACS Med Chem Lett
CHEMBL5148457	Toxicity	Inhibition of recombinant human DHFR expressed in Escherichia coli BL21 (DE3) cells using dihydrofolate as substrate in presence of NADPH by spectrophotometric analysis	Multitarget, Selective Compound Design Yields Potent Inhibitors of a Kinetoplastid Pteridine Reductase 1. — J Med Chem

Clinical trials (associated diseases)

300 trials via MONDO — disease-level, not drug-specific.

Trial	Phase	Status	Title
NCT00190697	PHASE4	COMPLETED	A Study of LY353381 (Arzoxifene) for Patients Who Benefitted From This Drug in Other Oncology Trials and Wished to Continue Treatment
NCT00719017	PHASE4	UNKNOWN	Upper Vaginectomy Versus Brachytherapy in Patients With Early Stage Endometrial Cancer Treated With Laparoscopic Surgery
NCT02543710	PHASE4	RECRUITING	Biomarker Guided Treatment in Gynaecological Cancer
NCT03349463	PHASE4	UNKNOWN	Evaluation of Fluciclovine Uptake in Patients With Cervical, Ovarian Epithelial or Endometrial Cancers.
NCT03752606	PHASE4	COMPLETED	Application of Tachosil During Lymphadenectomy
NCT05949424	PHASE4	UNKNOWN	OPTI - DOSE: Optimal Dosing of Oral Anticancer Drugs in Older Adults
NCT06049693	PHASE4	COMPLETED	Iron Prehabilitation in Endometrial Cancer
NCT06726291	PHASE4	RECRUITING	Akynzeo as Antiemetic Treatment in Patients With Endometrial Cancer
NCT06871787	PHASE4	NOT_YET_RECRUITING	Near-Infrared Fluorescence Imaging With Indocyanine Green to Evaluate Bowel Anastomoses in Gynecologic Oncology Surgery
NCT07281547	PHASE4	NOT_YET_RECRUITING	Low Dose Aspirin to Lower Inflammation and Prevent Endometrial Cancer in Postmenopausal Women With Non-atrophic Endometrial Changes and Pain
NCT07462663	PHASE4	NOT_YET_RECRUITING	SHAPE-ENDO: Multimodal Pre-Surgical Optimization in Patients With Obesity and Early-Stage Endometrial Cancer (Phase 1)
NCT00002459	PHASE3	COMPLETED	Radiation Therapy or No Further Treatment Following Surgery in Treating Patients With Cancer of the Uterus
NCT00002493	PHASE3	COMPLETED	Radiation Therapy Compared With Combination Chemotherapy in Treating Patients With Advanced Endometrial Cancer
NCT00002641	PHASE3	COMPLETED	Surgery With or Without Chemotherapy in Treating Patients With Soft Tissue Sarcoma
NCT00002764	PHASE3	COMPLETED	Surgery With or Without Combination Chemotherapy in Treating Patients With Lung Metastases From Soft Tissue Sarcoma
NCT00002920	PHASE3	COMPLETED	S9630, Medroxyprogesterone in Treating Women With Breast Cancer
NCT00002976	PHASE3	TERMINATED	Estrogen Replacement Therapy in Treating Women With Early-Stage Endometrial Cancer
NCT00003267	PHASE3	COMPLETED	Pelvic Drains After Radical Hysterectomy in Treating Patients With Uterine, Cervical, or Vaginal Cancer
NCT00003691	PHASE3	COMPLETED	Combination Chemotherapy With or Without G-CSF in Treating Patients With Stage III, Stage IV, or Recurrent Endometrial Cancer
NCT00003749	PHASE3	COMPLETED	Surgery With or Without Lymphadenectomy and Radiation Therapy in Treating Patients With Endometrial Cancer
NCT00005583	PHASE3	COMPLETED	Radiation Therapy With or Without Chemotherapy in Treating Patients With High-Risk Endometrial Cancer
NCT00006027	PHASE3	COMPLETED	Comparison of Radiation Therapy With or Without Combination Chemotherapy Following Surgery in Treating Patients With Stage I or Stage II Endometrial Cancer
NCT00016341	PHASE3	TERMINATED	Combination Chemotherapy Compared With Hormone Therapy in Treating Patients With Recurrent, Stage III, or Stage IV Endometrial Cancer
NCT00033605	PHASE3	COMPLETED	Octreotide in Preventing Diarrhea in Patients Who Are Undergoing Radiation Therapy to the Pelvis
NCT00096408	PHASE3	COMPLETED	Laparoscopic Approach to Cancer of the Endometrium
NCT00245050	PHASE3	COMPLETED	Pyridoxine in Preventing Hand-Foot Syndrome in Patients Who Are Receiving Liposomal Doxorubicin for Cancer
NCT00376844	PHASE3	COMPLETED	External-Beam Radiation Therapy Compared With Vaginal Brachytherapy After Surgery for Stage I Endometrial Cancer
NCT00411138	PHASE3	UNKNOWN	Randomized Trial of Radiation Therapy With or Without Chemotherapy for Endometrial Cancer
NCT00566644	PHASE3	TERMINATED	Intrauterine Levonorgestrel and Observation or Observation Alone in Preventing Atypical Endometrial Hyperplasia and Endometrial Cancer in Women With Hereditary Non-Polyposis Colorectal Cancer or Lynch Syndrome
NCT00883116	PHASE3	TERMINATED	A Study of Ixabepilone as Second-line Therapy for Locally Advanced, Recurrent, or Metastatic Endometrial Cancer
NCT01087268	PHASE3	UNKNOWN	Hyperbaric Oxygen Therapy in Treating Long-Term Gastrointestinal Adverse Effects Caused by Radiation Therapy in Patients With Pelvic Cancer
NCT01470677	PHASE3	COMPLETED	Tachosil for the Prevention of Symptomatic Lymph Cysts
NCT01672892	PHASE3	COMPLETED	Standard Versus Intensity-Modulated Pelvic Radiation Therapy in Treating Patients With Endometrial or Cervical Cancer
NCT01767155	PHASE3	COMPLETED	Zoptarelin Doxorubicin (AEZS 108) as Second Line Therapy for Endometrial Cancer
NCT02584478	PHASE3	UNKNOWN	Phase 1/2a/3 Evaluation of Adding AL3818 to Standard Platinum-Based Chemotherapy in Subjects With Recurrent or Metastatic Endometrial, Ovarian, Fallopian, Primary Peritoneal or Cervical Carcinoma (AL3818-US-002)
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