MAP3K8

Summary

MAP3K8 (mitogen-activated protein kinase kinase kinase 8, HGNC:6860) is a protein-coding gene on chromosome 10p11.23, encoding Mitogen-activated protein kinase kinase kinase 8 (P41279). Required for lipopolysaccharide (LPS)-induced, TLR4-mediated activation of the MAPK/ERK pathway in macrophages, thus being critical for production of the pro-inflammatory cytokine TNF (TNF) during immune responses.

This gene is an oncogene that encodes a member of the serine/threonine protein kinase family. The encoded protein localizes to the cytoplasm and can activate both the MAP kinase and JNK kinase pathways. This protein was shown to activate IkappaB kinases, and thus induce the nuclear production of NF-kappaB. This protein was also found to promote the production of TNF-alpha and IL-2 during T lymphocyte activation. This gene may also utilize a downstream in-frame translation start codon, and thus produce an isoform containing a shorter N-terminus. The shorter isoform has been shown to display weaker transforming activity. Alternate splicing results in multiple transcript variants that encode the same protein.

Source: NCBI Gene 1326 — RefSeq curated summary.

At a glance

• GWAS associations: 6
• Clinical variants (ClinVar): 280 total
• Phenotypes (HPO): 5
• Druggable target: yes — 1 molecules with ChEMBL bioactivity
• MANE Select transcript: NM_005204

Identifiers

Gene identifiers

Field	Value
HGNC ID	HGNC:6860
Approved symbol	MAP3K8
Name	mitogen-activated protein kinase kinase kinase 8
Location	10p11.23
Locus type	gene with protein product
Status	Approved
Aliases	Tpl-2, EST, c-COT, MEKK8
Ensembl gene	ENSG00000107968
Ensembl biotype	protein_coding
OMIM	191195
Entrez	1326

Gene structure

Transcript identifiers

Ensembl transcripts: 18 — 17 protein_coding, 1 retained_intron

ENST00000263056, ENST00000375321, ENST00000375322, ENST00000413724, ENST00000415139, ENST00000430603, ENST00000542547, ENST00000897693, ENST00000897694, ENST00000897695, ENST00000919546, ENST00000919547, ENST00000971341, ENST00000971342, ENST00000971343, ENST00000971344, ENST00000971345, ENST00000971346

RefSeq mRNA: 3 — MANE Select: NM_005204 NM_001244134, NM_001320961, NM_005204

CCDS: CCDS7166

Canonical transcript exons

ENST00000263056 — 9 exons

Exon	Start	End
ENSE00000816397	30458084	30458236
ENSE00000816399	30438916	30439274
ENSE00000985785	30451638	30451744
ENSE00001150501	30459255	30459501
ENSE00001150513	30450258	30450519
ENSE00001150525	30437176	30437406
ENSE00001223574	30434184	30434378
ENSE00001466718	30460706	30461833
ENSE00003628545	30447782	30447949

Expression profiles

Bgee: expression breadth ubiquitous, 249 present calls, max score 97.28.

FANTOM5 (CAGE): breadth ubiquitous, TPM avg 42.2861 / max 1749.2789, expressed in 1727 samples.

FANTOM5 promoters (8 alternative TSS)

Promoter ID	TPM avg	Samples expressed
104542	25.6590	1608
104541	8.3648	1110
104543	4.1068	1031
104544	2.5674	589
104540	1.1076	552
104545	0.2088	61
104547	0.1628	58
104546	0.1089	47

Top tissues by expression

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

Tissue	Anatomy ID	Expression score	Quality
mucosa of stomach	UBERON:0001199	97.28	gold quality
mucosa of paranasal sinus	UBERON:0005030	96.26	gold quality
left uterine tube	UBERON:0001303	95.37	gold quality
upper lobe of left lung	UBERON:0008952	95.37	gold quality
germinal epithelium of ovary	UBERON:0001304	95.31	gold quality
omental fat pad	UBERON:0010414	94.91	gold quality
peritoneum	UBERON:0002358	94.82	gold quality
monocyte	CL:0000576	94.58	gold quality
olfactory segment of nasal mucosa	UBERON:0005386	94.46	gold quality
upper lobe of lung	UBERON:0008948	94.42	gold quality
calcaneal tendon	UBERON:0003701	94.20	gold quality
mononuclear cell	CL:0000842	94.00	gold quality
nerve	UBERON:0001021	93.98	gold quality
tibial nerve	UBERON:0001323	93.98	gold quality
adipose tissue of abdominal region	UBERON:0007808	93.76	gold quality
right lung	UBERON:0002167	93.69	gold quality
leukocyte	CL:0000738	93.60	gold quality
gall bladder	UBERON:0002110	93.32	gold quality
parietal pleura	UBERON:0002400	93.05	gold quality
ascending aorta	UBERON:0001496	93.04	gold quality
thoracic aorta	UBERON:0001515	93.00	gold quality
left coronary artery	UBERON:0001626	92.28	gold quality
descending thoracic aorta	UBERON:0002345	92.15	gold quality
granulocyte	CL:0000094	91.90	gold quality
skin of abdomen	UBERON:0001416	91.88	gold quality
pleura	UBERON:0000977	91.82	gold quality
spleen	UBERON:0002106	91.35	gold quality
aorta	UBERON:0000947	91.08	gold quality
subcutaneous adipose tissue	UBERON:0002190	90.99	gold quality
vermiform appendix	UBERON:0001154	90.79	gold quality

Single-cell (SCXA)

Detected in 10 experiment(s), a significant marker in 9.

Experiment	Marker?	Max mean expression
E-MTAB-8884	yes	804.43
E-HCAD-4	yes	49.58
E-CURD-122	yes	33.95
E-MTAB-6678	yes	21.95
E-CURD-46	yes	18.94
E-CURD-88	yes	16.82
E-MTAB-9467	yes	12.92
E-HCAD-10	yes	7.25
E-MTAB-7606	no	566.99
E-ANND-3	no	0.00

Regulation

Is transcription factor: no

Upstream regulators (CollecTRI, top): AP1, BCL11B, JUN, NFKB1, NFKB, TFAP2A

miRNA regulators (miRDB)

93 targeting MAP3K8, 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-5011-5P	100.00	83.46	5820
HSA-MIR-1277-5P	100.00	73.95	5056
HSA-MIR-190A-3P	100.00	80.35	5520
HSA-LET-7A-3P	100.00	74.03	3932
HSA-LET-7B-3P	100.00	74.08	3913
HSA-LET-7F-1-3P	100.00	74.02	3928
HSA-MIR-98-3P	100.00	74.08	3907
HSA-MIR-3163	100.00	77.23	8605
HSA-MIR-3662	99.99	73.82	5684
HSA-MIR-4282	99.99	75.36	6408
HSA-MIR-520D-5P	99.98	73.34	4883
HSA-MIR-524-5P	99.98	73.43	4882
HSA-MIR-4789-5P	99.98	70.76	2721
HSA-MIR-5696	99.98	72.36	4487
HSA-MIR-146A-5P	99.96	68.93	988
HSA-MIR-146B-5P	99.96	69.13	977
HSA-LET-7C-3P	99.95	73.42	2862
HSA-MIR-144-3P	99.94	73.98	2698
HSA-MIR-7153-5P	99.94	68.89	1006
HSA-MIR-539-5P	99.93	70.30	2855
HSA-MIR-6768-5P	99.92	67.36	1942
HSA-MIR-8063	99.91	69.76	3146
HSA-MIR-106A-5P	99.90	73.94	2683
HSA-MIR-153-5P	99.89	73.86	6317
HSA-MIR-17-5P	99.89	73.83	2665
HSA-MIR-605-3P	99.88	69.22	1833
HSA-MIR-106B-5P	99.88	74.72	2795
HSA-MIR-20A-5P	99.88	74.76	2769
HSA-MIR-526B-3P	99.88	74.06	2587
HSA-MIR-20B-5P	99.88	74.01	2621

Literature-anchored findings (GeneRIF, showing 40)

• contributes to LMP1-induced NF-kB signaling downstream of TRAF2 (PMID:11932422)
• suggest that the activation of different signaling pathways by Cot and other MAP3Ks may be regulated separately and may provide evidence for how such discrimination by one member of this kinase family occurs. (PMID:12138205)
• Tpl2 is inhibited by and is a partner of NFkappab p105 (PMID:12667451)
• hKSR-2, a new member of the KSR family, negatively regulates Cot-mediated MAP kinase and NF-kappaB pathway signaling (PMID:12975377)
• the COOH-terminal domain of wild-type Cot regulates its stability and kinase specific activity (PMID:14517305)
• optimal TPL-2 stability in vivo requires interaction with ABIN-2 as well as p105 (PMID:15169888)
• Data support a role for MAP3K8 activity in cellular transformation, but suggest that mutational activation of the gene is a rare event in lung cancer. (PMID:15287022)
• phosphorylation of Cot at Thr-290 is necessary but not sufficient for full kinase activity in the MEK/ERK pathway (PMID:15466476)
• Tpl2/Cot is overexpressed in large granular lymphocyte proliferative disorders but not other T-cell neoplasias. (PMID:15575964)
• These results indicate a distinction between TNF Receptor family members CD40 and TNFR1 in their utilization of MAP3Ks, and demonstrate TRAF-dependence of Tpl2 association with the CD40 receptor complex. (PMID:15670770)
• Our results using purified proteins indicate that p105 binding improves COT solubility and stability while down-regulating kinase activity, consistent with cellular data showing that p105 functions as an inhibitor of COT. (PMID:16087150)
• Data highlight the specific requirements for activation of the Cot-MKK1-ERK1/ERK2 pathway and provide evidence that Cot controls the functions of IL-1 that are mediated by ERK1/ERK2. (PMID:16371247)
• deregulated activation of Tpl2/Cot may occur in human cancer cells (PMID:16565081)
• Mutagenesis experiments indicated that COT activation is initiated by Thr290 phosphorylation catalysed by an IL-1-stimulated protein kinase distinct from IKKbeta, while Ser62 phosphorylation is an autophosphorylation event required for maximal activation. (PMID:16806191)
• MAP3K8 was found overexpressed in 30% of the endometrial carcinoma samples (PMID:17290588)
• TPL-2 is not targeted by mutation in diffuse large B cell lymphoma and myeloid leukemia (PMID:17324460)
• MAP3K8 and PRKCZ cooperate in the regulation of the transcriptional activity of NFATC2 through the phosphorylation of its amino-terminal domain. (PMID:17398070)
• the transforming ability of Cot results from the coordinated activation of histone H3, which ultimately converges on the regulation of the transcriptional activity of the c-fos promoter, followed by AP-1 transactivation activity (PMID:17724252)
• inhibition of Tpl2 in primary human cell types can decrease the production of TNFalpha and other pro-inflammatory mediators during inflammatory events (PMID:17848581)
• processing of pre-TNF alpha in LPS-stimulated macrophages is regulated by TPL2-mediated activation of ERK1 and ERK2 (PMID:18187448)
• Results describe the proteomics analysis of immunoprecipitated proteins associated with the oncogenic kinase Cot. (PMID:18319612)
• Cot was identified as a novel p65 interacting protein kinase. (PMID:18439422)
• Cot/Tpl2 did not function as a member of MAPK family, but as a promoter of renal cell apoptosis in ischemia reperfusion injury. (PMID:18518937)
• The increase in COX2 expression and the activation of Erk1/2 regulated by Cot are essential for the induction of cell migration. (PMID:18572386)
• the results from recent studies suggest that Tpl2/Cot and COX-2 could be prognostic factors in breast cancer [review] (PMID:18795070)
• These data underscore the role of Tpl2 as a regulator of T helper cell lineage decisions and demonstrate that Tpl2 has an important functional role in the regulation of Th1 responses. (PMID:19001140)
• Selected ‘Tpl2/Cot-YL ribozyme’ efficiently cleaves its target sequence in cis and in trans; furthermore, the ribozyme efficiently cleaves a longer target sequence of 54 nucleotides in trans, as well as the full-length mRNA. (PMID:19054068)
• Data support MAP3K8-induced activation of different MAPK signaling pathways in response to different profiles of shear stress, possibly as a consequence of shear-induced IL1B expression. (PMID:19272346)
• Cot protein, expressed in HEK293 cells and immunoprecipitated, was used in a peptide-based substrate screening assay. The results of this assay suggested that Polo-like kinase 1 (Plk1) was a substrate of Cot. (PMID:19804365)
• endogenous Tpl2 promotes efficient murine gammaherpesvirus 68 lytic replication through AP-1-dependent upregulation of RTA expression (PMID:19939924)
• MAP3 kinase COT1 is up-regulated by 1,25-dihydroxyvitamin D3 in parallel with activated c-jun during differentiation of human myeloid leukemia cells (PMID:20227498)
• Tensile strain and magnetic particle force application do not induce MAP3K8 and IL-1B differential gene expression in a similar manner to fluid shear stress in human mesenchymal stem cells. (PMID:20603871)
• Oncoprotein Cot1 represses kinase suppressors of Ras1/2 and 1,25-dihydroxyvitamin D3-induced differentiation of human acute myeloid leukemia cells. (PMID:20945381)
• identification of MAP3K8 (the gene encoding COT/Tpl2) as a MAPK pathway agonist that drives resistance to RAF inhibition in B-RAF(V600E) cell lines (PMID:21107320)
• TPL2 kinase plays a critical role in the promotion of androgen depletion-independent prostate cancer progression. (PMID:21267413)
• OPN knockdown chemosensitized MDA-MB-231 cells to CTX, which is dependent on p38 MAPK pathway activation (PMID:21539449)
• Cot protein is responsible for the constitutive Erk1/2 activation in the anaplastic large-cell lymphoma cells. (PMID:21741362)
• the role of Tpl2 in GPCR-mediated Ca(2+) signaling and cell migration (PMID:21868363)
• High TPL2 expression is associated with tumor progression. (PMID:23125217)
• Authors report constitutive activation of MAP3K8 kinase-dependent pathways that regulate the magnitude and extent of inflammatory activity of monocytes/macrophages within myeloma niches. (PMID:23252623)

Cross-species orthologs

3 orthologs

Organism	Symbol	Gene ID
danio_rerio	map3k8	ENSDARG00000061710
mus_musculus	Map3k8	ENSMUSG00000024235
rattus_norvegicus	Map3k8	ENSRNOG00000016378

Paralogs (35): MAP3K14 (ENSG00000006062), MAP4K3 (ENSG00000011566), MAP4K5 (ENSG00000012983), MAP2K3 (ENSG00000034152), SLK (ENSG00000065613), MAP4K4 (ENSG00000071054), STK10 (ENSG00000072786), PAK3 (ENSG00000077264), STRADB (ENSG00000082146), MAP3K1 (ENSG00000095015), STK4 (ENSG00000101109), PAK5 (ENSG00000101349), STK24 (ENSG00000102572), STK3 (ENSG00000104375), MAP4K1 (ENSG00000104814), MAP2K6 (ENSG00000108984), NEK4 (ENSG00000114904), STK25 (ENSG00000115694), NRK (ENSG00000123572), PAK4 (ENSG00000130669), STK26 (ENSG00000134602), TAOK3 (ENSG00000135090), PAK6 (ENSG00000137843), MINK1 (ENSG00000141503), PAK1 (ENSG00000149269), TAOK2 (ENSG00000149930), TNIK (ENSG00000154310), TAOK1 (ENSG00000160551), MAP4K2 (ENSG00000168067), OXSR1 (ENSG00000172939), MAP3K19 (ENSG00000176601), PAK2 (ENSG00000180370), SBK2 (ENSG00000187550), STK39 (ENSG00000198648), STRADA (ENSG00000266173)

Protein

Protein identifiers

Mitogen-activated protein kinase kinase kinase 8 — P41279 (reviewed: P41279)

Alternative names: Cancer Osaka thyroid oncogene, Proto-oncogene c-Cot, Serine/threonine-protein kinase cot, Tumor progression locus 2

All UniProt accessions (4): P41279, Q5T853, Q5T854, Q5T857

UniProt curated annotations — full annotation on UniProt →

Function. Required for lipopolysaccharide (LPS)-induced, TLR4-mediated activation of the MAPK/ERK pathway in macrophages, thus being critical for production of the pro-inflammatory cytokine TNF (TNF) during immune responses. Involved in the regulation of T-helper cell differentiation and IFNG expression in T-cells. Involved in mediating host resistance to bacterial infection through negative regulation of type I interferon (IFN) production. In vitro, activates MAPK/ERK pathway in response to IL1 in an IRAK1-independent manner, leading to up-regulation of IL8 and CCL4. Transduces CD40 and TNFRSF1A signals that activate ERK in B-cells and macrophages, and thus may play a role in the regulation of immunoglobulin production. May also play a role in the transduction of TNF signals that activate JNK and NF-kappa-B in some cell types. In adipocytes, activates MAPK/ERK pathway in an IKBKB-dependent manner in response to IL1B and TNF, but not insulin, leading to induction of lipolysis. Plays a role in the cell cycle. Isoform 1 shows some transforming activity, although it is much weaker than that of the activated oncogenic variant.

Subunit / interactions. Forms a ternary complex with NFKB1/p105 and TNIP2. Interacts with NFKB1; the interaction increases the stability of MAP3K8 but inhibits its MEK phosphorylation activity, whereas loss of interaction following LPS stimulation leads to its degradation. Interacts with CD40 and TRAF6; the interaction is required for ERK activation. Interacts with KSR2; the interaction inhibits ERK and NF-kappa-B activation.

Subcellular location. Cytoplasm.

Tissue specificity. Expressed in several normal tissues and human tumor-derived cell lines.

Post-translational modifications. Autophosphorylated. Isoform 1 undergoes phosphorylation mainly on Ser residues, and isoform 2 on both Ser and Thr residues. Phosphorylated on Thr-290; the phosphorylation is necessary but not sufficient for full kinase activity in vitro and for the dissociation of isoform 1 from NFKB1, leading to its degradation. Phosphorylated on Ser-400 by IKBKB; the phosphorylation is required for LPS-stimulated activation of the MAPK/ERK pathway in macrophages.

Induction. Up-regulated by IL12 in T-lymphocytes. Up-regulated in subcutaneous adipose tissue of obese individuals.

Miscellaneous. Can be converted to an oncogenic protein by proviral activation, leading to a C-terminally truncated protein with transforming activity.

Similarity. Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.

Isoforms (2)

UniProt ID	Names	Canonical?
P41279-1	1, 58 kDa	yes
P41279-2	2, 52 kDa

RefSeq proteins (3): NP_001231063, NP_001307890, NP_005195* (*=MANE)

Domains & families (InterPro)

ID	Name	Type
IPR000719	Prot_kinase_dom	Domain
IPR008271	Ser/Thr_kinase_AS	Active_site
IPR011009	Kinase-like_dom_sf	Homologous_superfamily
IPR017424	MAPKKK8	Family
IPR050538	MAP_kinase_kinase_kinase	Family

Pfam: PF00069

Catalyzed reactions (Rhea), 2 shown:

• L-seryl-[protein] + ATP = O-phospho-L-seryl-[protein] + ADP + H(+) (RHEA:17989)
• L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] + ADP + H(+) (RHEA:46608)

UniProt features (50 total): helix 15, strand 13, modified residue 5, turn 4, sequence variant 3, mutagenesis site 3, binding site 2, chain 1, domain 1, splice variant 1, sequence conflict 1, active site 1

Structure

Experimental structures (PDB)

3 structures.

PDB	Method	Resolution (Å)
4Y85	X-RAY DIFFRACTION	2.33
5IU2	X-RAY DIFFRACTION	2.7
4Y83	X-RAY DIFFRACTION	2.89

Predicted structure (AlphaFold)

Model	pLDDT	Fraction very-high
AF-P41279-F1	74.13	0.47

Functional residue map

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

Catalytic / active sites (1): 253 (proton acceptor)

Ligand- & substrate-binding residues (2): 144–152; 167

Post-translational modifications (5): 80, 141, 290, 400, 443

Mutagenesis-validated functional residues (3):

Position	Phenotype
290	loss of mek phosphorylation activity and almost abolished autophosphorylation activity. loss of il1-stimulated mek phosp
290	impaired mek phosphorylation and autophosphorylation activities. no effect on ksr2 binding.
290	loss of mek phosphorylation activity and almost abolished autophosphorylation activity. no effect on ksr2 binding.

Function

Pathways and Gene Ontology

Reactome pathways

29 pathways

ID	Pathway
R-HSA-389357	CD28 dependent PI3K/Akt signaling
R-HSA-5684264	MAP3K8 (TPL2)-dependent MAPK1/3 activation
R-HSA-1280215	Cytokine Signaling in Immune system
R-HSA-1280218	Adaptive Immune System
R-HSA-166016	Toll Like Receptor 4 (TLR4) Cascade
R-HSA-166058	MyD88:MAL(TIRAP) cascade initiated on plasma membrane
R-HSA-166166	MyD88-independent TLR4 cascade
R-HSA-168138	Toll Like Receptor 9 (TLR9) Cascade
R-HSA-168142	Toll Like Receptor 10 (TLR10) Cascade
R-HSA-168164	Toll Like Receptor 3 (TLR3) Cascade
R-HSA-168176	Toll Like Receptor 5 (TLR5) Cascade
R-HSA-168179	Toll Like Receptor TLR1:TLR2 Cascade
R-HSA-168181	Toll Like Receptor 7/8 (TLR7/8) Cascade
R-HSA-168188	Toll Like Receptor TLR6:TLR2 Cascade
R-HSA-168249	Innate Immune System
R-HSA-168256	Immune System
R-HSA-168898	Toll-like Receptor Cascades
R-HSA-181438	Toll Like Receptor 2 (TLR2) Cascade
R-HSA-388841	Regulation of T cell activation by CD28 family
R-HSA-389356	Co-stimulation by CD28
R-HSA-446652	Interleukin-1 family signaling
R-HSA-448424	Interleukin-17 signaling
R-HSA-449147	Signaling by Interleukins
R-HSA-450294	MAP kinase activation
R-HSA-9020702	Interleukin-1 signaling
R-HSA-937061	TRIF (TICAM1)-mediated TLR4 signaling
R-HSA-975138	TRAF6 mediated induction of NFkB and MAP kinases upon TLR7/8 or 9 activation
R-HSA-975155	MyD88 dependent cascade initiated on endosome
R-HSA-975871	MyD88 cascade initiated on plasma membrane

MSigDB gene sets: 488 (showing top): GSE45365_CD8A_DC_VS_CD11B_DC_IFNAR_KO_MCMV_INFECTION_DN, GSE18804_SPLEEN_MACROPHAGE_VS_BRAIN_TUMORAL_MACROPHAGE_UP, WAMUNYOKOLI_OVARIAN_CANCER_LMP_DN, GOBP_REGULATION_OF_CELL_ACTIVATION, CREL_01, REACTOME_INNATE_IMMUNE_SYSTEM, REACTOME_ADAPTIVE_IMMUNE_SYSTEM, REACTOME_CYTOKINE_SIGNALING_IN_IMMUNE_SYSTEM, GOBP_INFLAMMATORY_RESPONSE, TGCACTT_MIR519C_MIR519B_MIR519A, KEGG_MAPK_SIGNALING_PATHWAY, MODULE_64, GOBP_LYMPHOCYTE_COSTIMULATION, PID_NFAT_3PATHWAY, YAO_TEMPORAL_RESPONSE_TO_PROGESTERONE_CLUSTER_1

GO Biological Process (7): MAPK cascade (GO:0000165), protein phosphorylation (GO:0006468), T cell costimulation (GO:0031295), positive regulation of inflammatory response (GO:0050729), immune system process (GO:0002376), signal transduction (GO:0007165), intracellular signal transduction (GO:0035556)

GO Molecular Function (11): magnesium ion binding (GO:0000287), protein serine/threonine kinase activity (GO:0004674), MAP kinase kinase kinase activity (GO:0004709), ATP binding (GO:0005524), protein serine kinase activity (GO:0106310), nucleotide binding (GO:0000166), protein kinase activity (GO:0004672), protein binding (GO:0005515), kinase activity (GO:0016301), transferase activity (GO:0016740), metal ion binding (GO:0046872)

GO Cellular Component (2): cytoplasm (GO:0005737), cytosol (GO:0005829)

Reactome top-level categories

Rollup of top-12 pathways:

Category	Pathways
Toll-like Receptor Cascades	7
Immune System	3
Toll Like Receptor 4 (TLR4) Cascade	2
Toll Like Receptor 2 (TLR2) Cascade	2
Co-stimulation by CD28	1
MAP kinase activation	1
Interleukin-1 signaling	1
Toll Like Receptor TLR1:TLR2 Cascade	1
Toll Like Receptor TLR6:TLR2 Cascade	1
Innate Immune System	1
Adaptive Immune System	1
Regulation of T cell activation by CD28 family	1

GO top-level categories

Rollup of top GO terms by namespace:

Category	Terms
intracellular anatomical structure	2
protein kinase activity	2
cellular anatomical structure	2
intracellular signaling cassette	1
phosphorylation	1
protein modification process	1
lymphocyte costimulation	1
positive regulation of T cell activation	1
inflammatory response	1
positive regulation of defense response	1
positive regulation of response to external stimulus	1
regulation of inflammatory response	1
biological_process	1
cell communication	1
cellular process	1
signaling	1
regulation of cellular process	1
cellular response to stimulus	1
signal transduction	1
metal ion binding	1
MAPK cascade	1
protein serine/threonine kinase activity	1
adenyl ribonucleotide binding	1
purine ribonucleoside triphosphate binding	1
nucleoside phosphate binding	1
heterocyclic compound binding	1
kinase activity	1
phosphotransferase activity, alcohol group as acceptor	1
catalytic activity, acting on a protein	1
binding	1
transferase activity, transferring phosphorus-containing groups	1
catalytic activity	1
cation binding	1
cytoplasm	1

Protein interactions and networks

STRING

1761 interactions, top by confidence (×1000):

Protein A	Protein B	Partner UniProt	Score
MAP3K8	TNIP2	Q8NFZ5	915
MAP3K8	NFKB1	P19838	753
MAP3K8	IL6	P05231	713
MAP3K8	NFKB2	Q00653	640
MAP3K8	IL1B	P01584	609
MAP3K8	RELA	Q04206	590
MAP3K8	TLR4	O00206	575
MAP3K8	IKBKB	O14920	556
MAP3K8	CXCL8	P10145	547
MAP3K8	MYD88	P78397	543
MAP3K8	IRAK1	P51617	496
MAP3K8	CCL5	P13501	493
MAP3K8	CCL2	P13500	480
MAP3K8	CHUK	O15111	476
MAP3K8	HSP90AA1	P07900	466

IntAct

69 interactions, top by confidence:

A	B	Type	Score
MAP3K8	NFKB1	psi-mi:“MI:0914”(association)	0.930
MAP3K8	NFKB1	psi-mi:“MI:0915”(physical association)	0.930
NFKB1	MAP3K8	psi-mi:“MI:0407”(direct interaction)	0.930
NFKB1	MAP3K8	psi-mi:“MI:0915”(physical association)	0.930
TNIP2	MAP3K8	psi-mi:“MI:0915”(physical association)	0.770
MAP3K8	TNIP2	psi-mi:“MI:0915”(physical association)	0.770
HSP90AB1	MAP3K8	psi-mi:“MI:0915”(physical association)	0.660
MAP3K8	HSP90AB1	psi-mi:“MI:0915”(physical association)	0.660
PIN1	MAP3K8	psi-mi:“MI:0915”(physical association)	0.660
MAP3K8	PIN1	psi-mi:“MI:0915”(physical association)	0.660
MAP3K8	PIN1	psi-mi:“MI:0217”(phosphorylation reaction)	0.660
TNIP2	NFKB1	psi-mi:“MI:0914”(association)	0.650
MAP2K1	MAP3K8	psi-mi:“MI:0217”(phosphorylation reaction)	0.440
MAP3K8	MAP2K1	psi-mi:“MI:0915”(physical association)	0.400
STIP1	MAP3K8	psi-mi:“MI:0915”(physical association)	0.370

BioGRID (89): MAP3K8 (Affinity Capture-MS), NFKB1 (Two-hybrid), NFKB1 (Affinity Capture-MS), NFKB2 (Affinity Capture-MS), RELA (Affinity Capture-MS), TNIP2 (Affinity Capture-MS), MAP3K8 (Affinity Capture-MS), NFKB1 (Affinity Capture-MS), NFKB2 (Affinity Capture-MS), RELA (Affinity Capture-MS), MAP3K8 (Affinity Capture-MS), MAP3K8 (Affinity Capture-MS), TNIP2 (FRET), MAP3K8 (FRET), MAP3K8 (FRET)

ESM2 similar proteins: A0AUV4, A1A5Q6, A1A5R7, A2KF29, B1WAS2, C0HKC8, C0HKC9, D3ZML2, O60285, O74536, O88831, O88866, P41279, P51956, P57058, P97756, Q20443, Q2T9U5, Q5R7G9, Q5XHI9, Q60670, Q63562, Q641K5, Q66HE5, Q68UT7, Q6P431, Q6VZ17, Q7T0B0, Q7T0B1, Q7TNJ7, Q7TNL4, Q8BHI9, Q8BZN4, Q8C078, Q8C0N0, Q8C0V7, Q8C0X8, Q8CIP4, Q8IY84, Q8K4K4

Diamond homologs: A2BD05, A4PES0, A4QNA8, A8XJW8, B7XHR6, D2HHP1, D3ZBE5, E2RSS3, F1LP90, F4JTP5, O08648, O18209, O34507, O57473, P07527, P0C1S8, P28327, P30291, P32944, P41279, P47810, P49615, P51954, P51956, P51957, P54350, P54644, P57059, P59895, Q00535, Q02399, Q0WPH8, Q10GB1, Q1LX51, Q20085, Q2PQN9, Q39008, Q3SZW1, Q54E34, Q54F40

SIGNOR signaling

21 interactions.

A	Effect	B	Mechanism
MAP3K8	up-regulates	MAP2K1	phosphorylation
MAP3K8	up-regulates	MAP2K2	phosphorylation
MAP3K8	up-regulates	MAP2K4	phosphorylation
NFKB1	down-regulates	MAP3K8	binding
NfKb-p65/p50	down-regulates	MAP3K8	binding
MAP3K8	up-regulates	MEK1/2	phosphorylation
MAP3K8	“up-regulates activity”	MAP3K14	phosphorylation
AKT	“up-regulates activity”	MAP3K8	phosphorylation
TRAF6	“up-regulates activity”	MAP3K8
AKT1	“up-regulates activity”	MAP3K8	phosphorylation
MAP3K8	“up-regulates activity”	PLCB3	phosphorylation
MAP3K8	up-regulates	PLK1	phosphorylation
MAP3K8	“up-regulates activity”	ERK1/2
TLR4	“up-regulates activity”	MAP3K8

Enriched among interaction partners

Reactome pathways and GO biological processes over-represented among this gene’s 33 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
HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand	8	53.4×	5e-10
The role of GTSE1 in G2/M progression after G2 checkpoint	5	27.7×	5e-05
PKR-mediated signaling	5	24.3×	7e-05
L1CAM interactions	5	20.7×	1e-04
Cytokine Signaling in Immune system	6	8.4×	7e-04
Axon guidance	5	7.8×	2e-03
Cellular responses to stress	6	7.6×	1e-03
Cellular responses to stimuli	7	7.6×	4e-04

GO biological processes:

GO term	Partners	Fold	FDR
protein refolding	5	111.5×	3e-07
protein stabilization	5	11.9×	2e-03

Disease & clinical

Clinical variants and AI predictions

ClinVar

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

Classification	Count (floor)
Pathogenic	0
Likely pathogenic	0
Uncertain significance	139
Likely benign	107
Benign	23

Top pathogenic / likely-pathogenic (0)

SpliceAI

1189 predictions. Top by Δscore:

Variant	Effect	Δscore
10:30438911:CCTA:C	acceptor_loss	1.0000
10:30438912:CTAG:C	acceptor_loss	1.0000
10:30438913:TAG:T	acceptor_loss	1.0000
10:30438914:A:AC	acceptor_loss	1.0000
10:30438914:A:AG	acceptor_gain	1.0000
10:30438915:G:A	acceptor_loss	1.0000
10:30438915:G:GG	acceptor_gain	1.0000
10:30447770:ATTTT:A	acceptor_gain	1.0000
10:30447771:T:G	acceptor_gain	1.0000
10:30447774:T:A	acceptor_gain	1.0000
10:30447777:T:A	acceptor_gain	1.0000
10:30447778:GTA:G	acceptor_loss	1.0000
10:30447779:TA:T	acceptor_loss	1.0000
10:30447780:A:AG	acceptor_gain	1.0000
10:30447780:AG:A	acceptor_gain	1.0000
10:30447781:G:GA	acceptor_gain	1.0000
10:30447781:GG:G	acceptor_gain	1.0000
10:30447781:GGT:G	acceptor_gain	1.0000
10:30447781:GGTC:G	acceptor_gain	1.0000
10:30447781:GGTCA:G	acceptor_gain	1.0000
10:30447945:AACTG:A	donor_gain	1.0000
10:30447950:G:GG	donor_gain	1.0000
10:30447950:GTAT:G	donor_loss	1.0000
10:30447951:T:A	donor_loss	1.0000
10:30450256:A:AG	acceptor_gain	1.0000
10:30450257:G:GA	acceptor_gain	1.0000
10:30450375:G:GT	donor_gain	1.0000
10:30450517:AACG:A	donor_loss	1.0000
10:30450520:G:GG	donor_gain	1.0000
10:30451636:A:AG	acceptor_gain	1.0000

AlphaMissense

3077 scored. Top likely-pathogenic:

Variant	Protein change	am_pathogenicity
10:30447946:A:C	K167N	1.000
10:30447946:A:T	K167N	1.000
10:30450466:T:C	L238P	1.000
10:30450475:T:A	L241H	1.000
10:30450475:T:C	L241P	1.000
10:30450484:T:C	L244P	1.000
10:30450504:C:G	H251D	1.000
10:30450511:A:C	D253A	1.000
10:30450511:A:G	D253G	1.000
10:30450511:A:T	D253V	1.000
10:30450512:T:A	D253E	1.000
10:30450512:T:G	D253E	1.000
10:30450516:A:G	K255E	1.000
10:30450518:A:C	K255N	1.000
10:30450518:A:T	K255N	1.000
10:30451645:C:A	N258K	1.000
10:30451645:C:G	N258K	1.000
10:30451679:G:C	D270H	1.000
10:30451680:A:C	D270A	1.000
10:30451680:A:G	D270G	1.000
10:30451680:A:T	D270V	1.000
10:30451681:T:A	D270E	1.000
10:30451681:T:G	D270E	1.000
10:30451740:C:T	T290I	1.000
10:30458093:A:C	S295R	1.000
10:30458095:C:A	S295R	1.000
10:30458095:C:G	S295R	1.000
10:30458135:G:C	D309H	1.000
10:30458135:G:T	D309Y	1.000
10:30458189:T:A	W327R	1.000

dbSNP variants (sampled 300 via entrez): RS1000244406 (10:30436870 ACCC>A), RS1000316869 (10:30453217 T>C), RS1000422699 (10:30459919 T>G), RS1000475177 (10:30459615 T>C), RS1000511223 (10:30459979 A>T), RS1000563707 (10:30459756 CT>C), RS1000605101 (10:30452823 G>A), RS1000670407 (10:30453484 G>A), RS1000740807 (10:30453874 A>G), RS1000806545 (10:30436366 C>T), RS1000869272 (10:30435731 A>G), RS1001114541 (10:30448111 G>A), RS1001141089 (10:30441484 C>T), RS1001153428 (10:30453739 G>A), RS1001336828 (10:30441847 T>A,C)

Disease associations

OMIM: gene MIM:191195 | disease phenotypes:

GenCC curated gene-disease

Mondo (0):

Orphanet (0):

HPO phenotypes

5 total (5 of 5 shown, HPO-id order):

HPO	Term
HP:0000006	Autosomal dominant inheritance
HP:0001442	Typified by somatic mosaicism
HP:0006519	Alveolar cell carcinoma
HP:0030078	Lung adenocarcinoma
HP:0030358	Non-small cell lung carcinoma

GWAS associations

6 associations (top):

Study	Trait	p-value
GCST001725_104	Inflammatory bowel disease	6.000000e-11
GCST004131_85	Inflammatory bowel disease	3.000000e-12
GCST004132_70	Crohn’s disease	5.000000e-10
GCST004133_42	Ulcerative colitis	5.000000e-06
GCST008887_3	Systemising	7.000000e-07
GCST010043_12	Asthma	2.000000e-08

EFO canonical traits (1, from GWAS)

EFO ID	Trait name
EFO:0010221	systemising measurement

Drugs & pharmacology

Drug and pharmacology data

Is drug target: yes

ChEMBL targets (1): CHEMBL4899 (SINGLE PROTEIN)

Molecules with ChEMBL bioactivity

1 molecules (phase ≥1), by development phase (incl. off-target/promiscuous compounds). Patent mentions across the top 20 by phase: 23,148 (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
CHEMBL6246	ELLAGIC ACID	2	23,148

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

GtoPdb / IUPHAR curated pharmacology

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

Target class: enzyme — STE-unique family

Most potent curated ligand interactions (2 total), top 2:

Ligand	Action	Affinity	Parameter
MEK inhibitor II	Inhibition	7.3	pIC50
Tpl2 kinase inhibitor	Inhibition	7.3	pIC50

Binding affinities (BindingDB)

79 measured of 88 human assays (89 total across all organisms); most potent 50 below. Values come from heterogeneous assays and are not directly comparable.

Ligand	Measure	Value
substituted 6-aminoquinoline-3-carbonitrile, 34	IC50	1.6 nM
8-chloro-4-[(3-chloro-4-fluorophenyl)amino]-6-[(2H-1,2,3,4-tetrazol-5-ylmethyl)amino]quinoline-3-carbonitrile	IC50	2 nM
substituted 6-aminoquinoline-3-carbonitrile, 21	IC50	2 nM
substituted 6-aminoquinoline-3-carbonitrile, 33	IC50	2.2 nM
substituted 6-aminoquinoline-3-carbonitrile, 23	IC50	3 nM
substituted 6-aminoquinoline-3-carbonitrile, 20	IC50	4 nM
substituted 6-aminoquinoline-3-carbonitrile, 32	IC50	5.3 nM
substituted 6-aminoquinoline-3-carbonitrile, 17	IC50	7 nM
substituted 6-aminoquinoline-3-carbonitrile, 22	IC50	7.6 nM
substituted 6-aminoquinoline-3-carbonitrile, 29	IC50	8 nM
substituted 6-aminoquinoline-3-carbonitrile, 35	IC50	8.2 nM
substituted 6-aminoquinoline-3-carbonitrile, 36	IC50	9.1 nM
substituted 6-aminoquinoline-3-carbonitrile, 12	IC50	9.6 nM
substituted 6-aminoquinoline-3-carbonitrile, 14	IC50	10 nM
substituted 6-aminoquinoline-3-carbonitrile, 37	IC50	10 nM
substituted 6-aminoquinoline-3-carbonitrile, 26	IC50	13 nM
8-chloro-4-[(3-chloro-4-fluorophenyl)amino]-6-({[4-(propan-2-yl)-1H-imidazol-5-yl]methyl}amino)quinoline-3-carbonitrile	IC50	16 nM
8-chloro-4-[(3-chloro-4-fluorophenyl)amino]-6-{[(4-ethyl-1H-imidazol-5-yl)methyl]amino}quinoline-3-carbonitrile	IC50	16 nM
substituted 6-aminoquinoline-3-carbonitrile, 24	IC50	17 nM
substituted 6-aminoquinoline-3-carbonitrile, 31	IC50	18 nM
7-amino-4-(3-phenylphenyl)thieno[2,3-c]pyridine-2-carboxylic acid	IC50	20 nM
substituted 6-aminoquinoline-3-carbonitrile, 19	IC50	20 nM
substituted 6-aminoquinoline-3-carbonitrile, 25	IC50	20 nM
substituted 6-aminoquinoline-3-carbonitrile, 27	IC50	22 nM
substituted 6-aminoquinoline-3-carbonitrile, 30	IC50	22 nM
substituted 6-aminoquinoline-3-carbonitrile, 18	IC50	23 nM
8-chloro-4-[(3-chloro-4-fluorophenyl)amino]-6-{[(4-propyl-1H-imidazol-5-yl)methyl]amino}quinoline-3-carbonitrile	IC50	25 nM
substituted 6-aminoquinoline-3-carbonitrile, 28	IC50	28 nM
BMCL193485 Compound 2	IC50	30 nM
8-bromo-4-[(3-chloro-4-fluorophenyl)amino]-6-{[(1-methyl-1H-imidazol-4-yl)methyl]amino}quinoline-3-carbonitrile	IC50	30 nM
substituted 6-aminoquinoline-3-carbonitrile, 7	IC50	40 nM
substituted 6-aminoquinoline-3-carbonitrile, 13	IC50	40 nM
8-bromo-4-[(3-chloro-4-fluorophenyl)amino]-6-[({1-[2-(morpholin-4-yl)ethyl]-1H-imidazol-4-yl}methyl)amino]quinoline-3-carbonitrile	IC50	41 nM
8-bromo-4-[(3-chloro-4-fluorophenyl)amino]-6-({[1-(2-methanesulfonylethyl)-1H-imidazol-4-yl]methyl}amino)quinoline-3-carbonitrile	IC50	43 nM
4-(3-phenylphenyl)thieno[2,3-c]pyridine-2-carboxylic acid	IC50	50 nM
substituted 6-aminoquinoline-3-carbonitrile, 15	IC50	61 nM
5-[4-(3-phenylphenyl)thieno[2,3-c]pyridin-2-yl]-1H-1,2,3,4-tetrazole	IC50	80 nM
methyl 5-[({8-bromo-4-[(3-chloro-4-fluorophenyl)amino]-3-cyanoquinolin-6-yl}amino)methyl]-1H-imidazole-4-carboxylate	IC50	87 nM
8-bromo-4-[(3-chloro-4-fluorophenyl)amino]-6-{[(1-methyl-1H-pyrazol-5-yl)methyl]amino}quinoline-3-carbonitrile	IC50	98 nM
2-({8-chloro-4-[(3-chloro-4-fluorophenyl)amino]-3-cyanoquinolin-6-yl}amino)-N-(pyridin-3-yl)acetamide	IC50	110 nM
substituted 6-aminoquinoline-3-carbonitrile, 10	IC50	120 nM
8-chloro-4-[(3-chloro-4-fluorophenyl)amino]-6-{[2-(1H-imidazol-4-yl)ethyl]amino}quinoline-3-carbonitrile	IC50	130 nM
8-bromo-4-[(3-chloro-4-fluorophenyl)amino]-6-({[4-(propan-2-yl)-1H-imidazol-5-yl]methyl}amino)quinoline-3-carbonitrile	IC50	130 nM
8-chloro-4-[(3-chloro-4-fluorophenyl)amino]-6-[({4-[3-(dimethylamino)propyl]-1H-imidazol-5-yl}methyl)amino]quinoline-3-carbonitrile	IC50	130 nM
5-[4-(4-phenylphenoxy)thieno[2,3-c]pyridin-2-yl]-1H-1,2,3,4-tetrazole	IC50	130 nM
4-[(4-phenylphenyl)amino]thieno[2,3-c]pyridine-2-carboxylic acid	IC50	140 nM
substituted 6-aminoquinoline-3-carbonitrile, 11	IC50	140 nM
4-[(3-chloro-4-fluorophenyl)amino]-8-acetyl-6-[(1H-imidazol-4-ylmethyl)amino]quinoline-3-carbonitrile	IC50	156 nM
8-chloro-4-[(3-chloro-4-fluorophenyl)amino]-6-{[(6-methylpyridin-2-yl)methyl]amino}quinoline-3-carbonitrile	IC50	170 nM
substituted 6-aminoquinoline-3-carbonitrile, 9	IC50	170 nM

ChEMBL bioactivities

242 potent at pChembl≥5 of 268 total, top 50 by pChembl (potency: 10 = 0.1 nM, 6 = 1 µM).

pChembl	Type	Value	Unit	Molecule
8.80	IC50	1.6	nM	CHEMBL527026
8.70	IC50	2	nM	CHEMBL371658
8.70	IC50	2	nM	CHEMBL230117
8.70	IC50	2	nM	CHEMBL498071
8.66	IC50	2.2	nM	CHEMBL498029
8.52	IC50	3	nM	CHEMBL219682
8.52	IC50	3	nM	CHEMBL221221
8.52	IC50	3	nM	CHEMBL482863
8.40	IC50	4	nM	CHEMBL3912476
8.40	IC50	4	nM	CHEMBL498070
8.28	IC50	5.3	nM	CHEMBL498028
8.22	IC50	6	nM	CHEMBL199210
8.22	IC50	6	nM	CHEMBL3939592
8.15	IC50	7	nM	CHEMBL495867
8.12	IC50	7.6	nM	CHEMBL498072
8.10	IC50	8	nM	CHEMBL496595
8.09	IC50	8.2	nM	CHEMBL495617
8.05	IC50	9	nM	CHEMBL3927501
8.05	IC50	9	nM	CHEMBL3890505
8.04	IC50	9.1	nM	CHEMBL523059
8.02	IC50	9.6	nM	CHEMBL495584
8.00	IC50	10	nM	CHEMBL241988
8.00	IC50	10	nM	CHEMBL491228
8.00	IC50	10	nM	CHEMBL501709
8.00	IC50	10	nM	CHEMBL498222
7.92	IC50	12	nM	CHEMBL199230
7.92	IC50	12	nM	CHEMBL436817
7.89	IC50	13	nM	CHEMBL483685
7.85	IC50	14	nM	CHEMBL219628
7.80	IC50	16	nM	CHEMBL3899461
7.80	IC50	16	nM	CHEMBL397986
7.80	IC50	16	nM	CHEMBL230554
7.80	IC50	16	nM	CHEMBL242852
7.80	IC50	16	nM	CHEMBL199230
7.77	IC50	17	nM	CHEMBL482864
7.75	IC50	18	nM	CHEMBL230232
7.75	IC50	18	nM	CHEMBL395573
7.75	IC50	18	nM	CHEMBL499853
7.72	IC50	19	nM	CHEMBL220037
7.70	IC50	20	nM	CHEMBL3910848
7.70	IC50	20	nM	CHEMBL220037
7.70	IC50	20	nM	CHEMBL425834
7.70	IC50	20	nM	CHEMBL242206
7.70	IC50	20	nM	CHEMBL436817
7.70	IC50	20	nM	CHEMBL245212
7.70	IC50	20	nM	CHEMBL476442
7.70	IC50	20	nM	CHEMBL483484
7.70	IC50	20	nM	CHEMBL524638
7.66	IC50	22	nM	CHEMBL3918552
7.66	IC50	22	nM	CHEMBL524069

PubChem BioAssay actives

328 with measured affinity, of 569 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
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(1-ethylpiperidin-4-yl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0016	uM
4-(4-benzylanilino)-6-(2-morpholin-4-ylethylamino)-1,7-naphthyridine-3-carbonitrile	257484: Inhibitory activity against human Tpl2 kinase via quantification of MEK phosphorylation	ic50	0.0020	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-(2H-tetrazol-5-ylmethylamino)quinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0020	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(pyridin-2-ylmethyl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0020	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(1-methylpiperidin-4-yl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0022	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[(1-pyridin-4-yltriazol-4-yl)methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0030	uM
4-(3-chloro-4-fluoroanilino)-6-[(5-methyl-1H-imidazol-4-yl)methylamino]quinoline-3-carbonitrile	276619: Inhibition of Tpl2 kinase by ELISA	ic50	0.0030	uM
4-(3-chloro-4-fluoroanilino)-6-[(1,5-dimethylimidazol-4-yl)methylamino]quinoline-3-carbonitrile	276619: Inhibition of Tpl2 kinase by ELISA	ic50	0.0030	uM
N-[(2R)-1-morpholin-4-ylpropan-2-yl]-6-(8-pyridin-4-ylimidazo[4,5-c][1,7]naphthyridin-1-yl)quinolin-2-amine	1316045: Inhibition of human COT (66 to 395 residues) expressed in Sf21 cells using 5-Fluo-Ahx-AGAGSGQLIDSNleANSFVGTR-NH2 as substrate after 60 mins by caliper microfluidic mobility shift assay	ic50	0.0040	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(pyridin-4-ylmethyl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0040	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[(1-piperidin-4-yltriazol-4-yl)methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0053	uM
N-(1-morpholin-4-ylpropan-2-yl)-6-(8-pyridin-4-ylimidazo[4,5-c][1,7]naphthyridin-1-yl)quinolin-2-amine	1316045: Inhibition of human COT (66 to 395 residues) expressed in Sf21 cells using 5-Fluo-Ahx-AGAGSGQLIDSNleANSFVGTR-NH2 as substrate after 60 mins by caliper microfluidic mobility shift assay	ic50	0.0060	uM
6-(2-morpholin-4-ylethylamino)-4-(4-phenylsulfanylanilino)-1,7-naphthyridine-3-carbonitrile	257484: Inhibitory activity against human Tpl2 kinase via quantification of MEK phosphorylation	ic50	0.0060	uM
2-[4-[[[8-bromo-4-(3-chloro-4-fluoroanilino)-3-cyanoquinolin-6-yl]amino]methyl]triazol-1-yl]acetic acid	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0070	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[(1-pyridin-3-yltriazol-4-yl)methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0076	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(2-morpholin-4-ylethyl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0080	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(1-propylpiperidin-4-yl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0082	uM
N-(1-morpholin-4-ylpropan-2-yl)-6-(8-pyridin-4-ylimidazo[4,5-c][1,7]naphthyridin-1-yl)-1,3-benzothiazol-2-amine	1316045: Inhibition of human COT (66 to 395 residues) expressed in Sf21 cells using 5-Fluo-Ahx-AGAGSGQLIDSNleANSFVGTR-NH2 as substrate after 60 mins by caliper microfluidic mobility shift assay	ic50	0.0090	uM
N-(1-morpholin-4-ylpropan-2-yl)-6-(8-phenylimidazo[4,5-c][1,7]naphthyridin-1-yl)-1,3-benzothiazol-2-amine	1316045: Inhibition of human COT (66 to 395 residues) expressed in Sf21 cells using 5-Fluo-Ahx-AGAGSGQLIDSNleANSFVGTR-NH2 as substrate after 60 mins by caliper microfluidic mobility shift assay	ic50	0.0090	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(1-propan-2-ylpiperidin-4-yl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0091	uM
2-[4-[[[8-chloro-4-(3-chloro-4-fluoroanilino)-3-cyanoquinolin-6-yl]amino]methyl]triazol-1-yl]acetic acid	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0096	uM
8-bromo-4-(3-chloro-4-fluoroanilino)-6-(1H-imidazol-5-ylmethylamino)quinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0100	uM
4-[4-[2-(2H-tetrazol-5-yl)thieno[2,3-c]pyridin-4-yl]oxyphenyl]benzamide	348688: Inhibition of human recombinant COT by HTRF-based assay	ic50	0.0100	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(pyridin-3-ylmethyl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0100	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(1-cyclobutylpiperidin-4-yl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0100	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-(1H-imidazol-5-ylmethylamino)quinoline-3-carbonitrile	381480: Inhibition of human Tpl2 by ELISA	ic50	0.0120	uM
4-(3-chloro-4-fluoroanilino)-6-[[(1R)-1-phenylethyl]amino]-1,7-naphthyridine-3-carbonitrile	257484: Inhibitory activity against human Tpl2 kinase via quantification of MEK phosphorylation	ic50	0.0120	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-[2-(dimethylamino)ethyl]triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0130	uM
4-(3-chloro-4-fluoroanilino)-6-[[5-methyl-1-(2-morpholin-4-ylethyl)imidazol-4-yl]methylamino]quinoline-3-carbonitrile	276619: Inhibition of Tpl2 kinase by ELISA	ic50	0.0140	uM
N-(2-morpholin-4-ylethyl)-6-(8-phenylimidazo[4,5-c][1,7]naphthyridin-1-yl)-1,3-benzothiazol-2-amine	1316045: Inhibition of human COT (66 to 395 residues) expressed in Sf21 cells using 5-Fluo-Ahx-AGAGSGQLIDSNleANSFVGTR-NH2 as substrate after 60 mins by caliper microfluidic mobility shift assay	ic50	0.0160	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-(pyridin-3-ylmethylamino)quinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0160	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[(4-propan-2-yl-1H-imidazol-5-yl)methylamino]quinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0160	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[(4-ethyl-1H-imidazol-5-yl)methylamino]quinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0160	uM
6-[[1-(2-aminoethyl)triazol-4-yl]methylamino]-8-chloro-4-(3-chloro-4-fluoroanilino)quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0170	uM
8-bromo-4-(3-chloro-4-fluoroanilino)-6-(2H-triazol-4-ylmethylamino)quinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0180	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[(1-oxidopyridin-1-ium-2-yl)methylamino]quinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0180	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-[2-(1-ethylpiperidin-4-yl)ethyl]triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0180	uM
4-(3-chloro-4-fluoroanilino)-6-(1H-imidazol-5-ylmethylamino)quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0190	uM
1-quinolin-6-yl-8-thiophen-3-ylimidazo[4,5-c][1,7]naphthyridine	1316045: Inhibition of human COT (66 to 395 residues) expressed in Sf21 cells using 5-Fluo-Ahx-AGAGSGQLIDSNleANSFVGTR-NH2 as substrate after 60 mins by caliper microfluidic mobility shift assay	ic50	0.0200	uM
8-bromo-4-(3-chloro-4-fluoroanilino)-6-[[1-[3-(dimethylamino)propyl]triazol-4-yl]methylamino]quinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0200	uM
4-(3-chloro-4-fluoroanilino)-8-fluoro-6-(1H-imidazol-5-ylmethylamino)quinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0200	uM
4-(3-chloro-4-fluoroanilino)-6-(1H-imidazol-5-ylmethylamino)-8-methylsulfinylquinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0200	uM
7-amino-4-(3-phenylphenyl)thieno[2,3-c]pyridine-2-carboxylic acid	1798863: COT HTRF Assay from Article 10.1016/j.bmcl.2009.01.088: “Identification of a selective thieno[2,3-c]pyridine inhibitor of COT kinase and TNF-alpha production.”	ic50	0.0200	uM
8-bromo-4-(3-chloro-4-fluoroanilino)-6-[[1-(pyridin-3-ylmethyl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0200	uM
6-[[1-(3-aminopropyl)triazol-4-yl]methylamino]-8-chloro-4-(3-chloro-4-fluoroanilino)quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0200	uM
N-(1-morpholin-4-ylpropan-2-yl)-6-(8-thiophen-2-ylimidazo[4,5-c][1,7]naphthyridin-1-yl)-1,3-benzothiazol-2-amine	1316045: Inhibition of human COT (66 to 395 residues) expressed in Sf21 cells using 5-Fluo-Ahx-AGAGSGQLIDSNleANSFVGTR-NH2 as substrate after 60 mins by caliper microfluidic mobility shift assay	ic50	0.0220	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-(2-piperidin-1-ylethyl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0220	uM
8-chloro-4-(3-chloro-4-fluoroanilino)-6-[[1-[2-(1-methylpyrrolidin-2-yl)ethyl]triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0220	uM
8-acetyl-4-(3-chloro-4-fluoroanilino)-6-(1H-imidazol-5-ylmethylamino)quinoline-3-carbonitrile	1798140: Tpl2/Cot Kinase Assay from Article 10.1021/jm070436q: “Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.”	ic50	0.0230	uM
8-bromo-4-(3-chloro-4-fluoroanilino)-6-[[1-(2-pyrrolidin-2-ylethyl)triazol-4-yl]methylamino]quinoline-3-carbonitrile	1799045: Tpl2/Cot ELISA Assay from Article 10.1016/j.bmcl.2009.05.009: “Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood.”	ic50	0.0230	uM

CTD chemical–gene interactions

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

Chemical	Actions (top 5)	PubMed papers
Valproic Acid	affects cotreatment, increases expression, affects expression, decreases expression	7
sodium arsenite	increases expression, increases stability, decreases expression	4
Benzo(a)pyrene	affects methylation, decreases methylation, increases expression	4
Cisplatin	decreases expression, affects expression, affects cotreatment, increases expression	3
Tetrachlorodibenzodioxin	increases expression	3
Tretinoin	increases expression	3
Aflatoxin B1	affects expression, decreases methylation, increases methylation	3
Particulate Matter	decreases expression, increases abundance, increases expression	3
S-(1,2-dichlorovinyl)cysteine	affects response to substance, increases expression, affects cotreatment, decreases expression	2
Decitabine	affects expression, decreases expression	2
Zoledronic Acid	increases expression	2
Arsenic	affects methylation, decreases expression, increases abundance	2
Doxorubicin	decreases expression, decreases response to substance	2
Methotrexate	decreases expression, increases expression	2
Phenylmercuric Acetate	affects cotreatment, increases expression	2
Tobacco Smoke Pollution	increases expression	2
Cyclosporine	decreases expression	2
Cadmium Chloride	decreases expression, increases expression	2
aristolochic acid I	decreases expression	1
3-((6-(2-methoxyphenyl)pyrimidin-4-yl)amino)phenyl)methane sulfonamide	decreases expression	1
Glupearl 19S	increases expression	1
TL8-506	affects cotreatment, increases expression	1
urushiol	increases expression	1
chloroacetaldehyde	decreases expression	1
ethylbenzene	increases expression	1
methylmercuric chloride	increases expression	1
triphenyl phosphate	affects expression	1
pirinixic acid	affects binding, increases activity, increases expression	1
sodium arsenate	decreases expression, increases abundance	1
2-methyl-4-isothiazolin-3-one	increases expression	1

ChEMBL screening assays

128 unique, capped per target: 126 binding, 2 functional

Representative assays (with source publication via chembl_document):

Assay ID	Type	Description	Source paper
CHEMBL1000656	Binding	Inhibition of Tpl2 kinase by ELISA	Selective inhibitors of tumor progression loci-2 (Tpl2) kinase with potent inhibition of TNF-alpha production in human whole blood. — Bioorg Med Chem Lett
CHEMBL855104	Functional	Inhibition of MEK phosphorylation in LPS-treated human monocytes	Inhibition of Tpl2 kinase and TNFalpha production with quinoline-3-carbonitriles for the treatment of rheumatoid arthritis. — Bioorg Med Chem Lett

Cellosaurus cell lines

6 cell lines: 6 cancer cell line

First 10 cell lines (id-ordered, not curated):

Cellosaurus	Name	Category	Sex
CVCL_B1WL	Abcam HeLa MAP3K8 KO	Cancer cell line	Female
CVCL_D7UE	Ubigene A-549 MAP3K8 KO	Cancer cell line	Male
CVCL_E0HC	Ubigene HeLa MAP3K8 KO	Cancer cell line	Female
CVCL_SW74	HAP1 MAP3K8 (-) 1	Cancer cell line	Male
CVCL_SW75	HAP1 MAP3K8 (-) 2	Cancer cell line	Male
CVCL_SW76	HAP1 MAP3K8 (-) 3	Cancer cell line	Male

Clinical trials (associated diseases)

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