Wnt signaling plays important roles in carcinogenesis and embryonic development. TNIK is a serine/threonine kinase that functions as an activator of the Wnt signaling pathway. TNIK is recruited to promoters of Wnt target genes and required to activate their expression. TCF4 is a substrate of the TNIK enzyme. TNIK phosphorylates TCF4 and regulates its transcriptional activity. TNIK together with NEDD4 and RAP2A form a signaling complex which regulates neuronal dendrite extension and arborization during development. TNIK may also play a role in cytoskeletal rearrangements and regulate cell spreading[3].
NCB-0846 is a TNIK inhibitor. The inhibitory IC50 of NCB-0846 against TNIK kinase was 21 nM[4]. In an experiment system of recombinant TCF4 and TNIK proteins, NCB-0846 induced faster migration of TCF4 phosphorylated by TNIK within a concentration range of 0.1 - 0.3 μM and completely inhibited the phosphorylation of TCF4 at a concentration of 3 μM . Furthermore, NCB-0846 blocked the autophosphorylation of TNIK in hct116 cells. At the concentrations of 3 μM or 10 μM, NCB-0846 reduced the expression of the Wnt target genes AXIN2 and MYC as well as that of TNIK in hct116 cells. NCB-0846 also reduced the expression of TNIK, AXIN2 and cMYC at the protein level at the concentration range of 1 – 10 μM in hct116 and DLD-1 cells[4]. According to another report, NCB-0846 at the concentrations of 1 μM or 3 μM reduced colony formation of cells derived from patients of prostate cancer[5].
In a hct116 xenograft model established in balb/c nude mice, NCB-0846 was administrated to the mice orally at the doses of 40 mg/kg or 80 mg/kg bid. Both doses of NCB-0846 suppressed tumor growth[4]. In a Wnt-driven tumorigenesis model utilizing Apcmin/+ mice, 3 doses of NCB-0846, including 22.5 mg/kg, 45 mg/kg, 90 mg/kg were administrated. NCB-0846 dose dependently reduced the multiplicity and dimensions of tumours that developed in the small intestine of mice[4].
NCB-0846 细胞实验
Cell Line
Concentration
Treated Time
Description
References
HEK293
0.1-0.3 µM
24 hours
Inhibition of TCF/LEF transcriptional activity
Nat Commun. 2016 Aug 26;7:12586
DLD-1
0.1-0.3 µM
24 hours
Inhibition of TCF/LEF transcriptional activity
Nat Commun. 2016 Aug 26;7:12586
HCT116
0.1-0.3 µM
24 hours
Inhibition of TCF/LEF transcriptional activity
Nat Commun. 2016 Aug 26;7:12586
PDX cells
0–10 µM
12 days
To assess the effect of NCB-0846 on colony formation, results showed that NCB-0846 significantly reduced colony formation.
Neoplasia. 2019 Apr;21(4):389-400
PDX cells
0–10 µM
1-5 days
To assess the effect of NCB-0846 on cell viability, results showed that NCB-0846 significantly reduced cell viability.
Neoplasia. 2019 Apr;21(4):389-400
PDX cells
0–10 µM
24 hours
To assess the effect of NCB-0846 on TNIK and TNIK (pS764) expression, results showed that NCB-0846 significantly decreased TNIK and TNIK (pS764) levels.
Neoplasia. 2019 Apr;21(4):389-400
MC3T3-E1 osteoblasts
0.2 µM
24 hours
To study the effect of NCB-0846 on collagen deposition in osteoblasts. Results showed that NCB-0846 treatment did not limit collagen deposition.
Hepatol Commun. 2022 Mar;6(3):593-609
Primary human hepatic stellate cells
0.1 µM
24 hours
To evaluate the inhibition of TNIK kinase activity by NCB-0846 and its effect on procollagen I secretion. It was found that NCB-0846 treatment reduced secretion of collagen I and fibronectin without affecting procollagen I transcription.
Hepatol Commun. 2022 Mar;6(3):593-609
LX-2 cells
0.1 µM
24 hours
To investigate the inhibitory effect of NCB-0846 on TNIK kinase activity and its impact on procollagen I secretion. Results showed that NCB-0846 treatment limited TNIK autophosphorylation and reduced secretion of collagen I and fibronectin.
Hepatol Commun. 2022 Mar;6(3):593-609
PDX cells
0–10 µM
4 weeks
To assess the effect of NCB-0846 on anchorage-independent growth in soft agar, results showed that NCB-0846 significantly reduced colony number and size.
Neoplasia. 2019 Apr;21(4):389-400
LUDLU1
300 nM
48 hours
To evaluate the radiosensitizing effect of NCB-0846 on TNIKlow LSCC cells, results showed that NCB-0846 did not significantly reduce the survival of TNIKlow cells.
Mol Cancer Ther. 2024 Apr 27;23(8):1201–11
KNS62
300 nM
48 hours
To evaluate the radiosensitizing effect of NCB-0846 on TNIKlow LSCC cells, results showed that NCB-0846 did not significantly reduce the survival of TNIKlow cells.
Mol Cancer Ther. 2024 Apr 27;23(8):1201–11
SW900
300 nM
48 hours
To evaluate the radiosensitizing effect of NCB-0846 on TNIKhigh LSCC cells, results showed that pretreatment with NCB-0846 significantly reduced the survival of TNIKhigh cells.
Mol Cancer Ther. 2024 Apr 27;23(8):1201–11
H520
300 nM
48 hours
To evaluate the radiosensitizing effect of NCB-0846 on TNIKhigh LSCC cells, results showed that pretreatment with NCB-0846 significantly reduced the survival of TNIKhigh cells.
Mol Cancer Ther. 2024 Apr 27;23(8):1201–11
LK2
300 nM
48 hours
To evaluate the radiosensitizing effect of NCB-0846 on TNIKhigh LSCC cells, results showed that pretreatment with NCB-0846 significantly reduced the survival of TNIKhigh cells.
Mol Cancer Ther. 2024 Apr 27;23(8):1201–11
NCI-H2228 cells
1 µM
48 hours
NCB-0846 modestly inhibited TGF β1-induced EMT in H2228 cells, but the effect on EMT markers was not obvious.
Br J Cancer. 2021 Jan;124(1):228-236
A549 cells
1 µM
48 hours
NCB-0846 inhibited TGF β1-induced EMT in A549 cells, restored E-cadherin expression, and suppressed mesenchymal markers vimentin and N-cadherin.
Br J Cancer. 2021 Jan;124(1):228-236
Aska
>2.0 µM
72 hours
Evaluate the inhibitory effect of NCB-0846 on Aska cells, results showed IC50 exceeding 2.0 µM, cells were insensitive to the drug
Cancers (Basel). 2020 May 16;12(5):1258
Yamato
767 nM
72 hours
Evaluate the inhibitory effect of NCB-0846 on Yamato cells, results showed IC50 of 767 nM, significantly inhibiting cell viability
Cancers (Basel). 2020 May 16;12(5):1258
SYO-1
356 nM
72 hours
Evaluate the inhibitory effect of NCB-0846 on SYO-1 cells, results showed IC50 of 356 nM, significantly inhibiting cell viability
Cancers (Basel). 2020 May 16;12(5):1258
HS-SY-II
339 nM
72 hours
Evaluate the inhibitory effect of NCB-0846 on HS-SY-II cells, results showed IC50 of 339 nM, significantly inhibiting cell viability
Cancers (Basel). 2020 May 16;12(5):1258
NCB-0846 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
C57BL/6J mice
CCl4-induced liver fibrosis model
Intraperitoneal injection
10 mg/kg
5 times per week for 4 weeks
To evaluate the inhibitory effect of NCB-0846 on liver fibrosis in vivo. Results showed that NCB-0846 treatment reduced CCl4-induced fibrosis and decreased expression of collagen I and fibronectin.
Hepatol Commun. 2022 Mar;6(3):593-609
NOD-scid IL2Rgammanull mice
Subcutaneous xenograft model
Oral gavage
100 mg/kg
Administered on days 1, 3, and 5, lasting for 1 week
To evaluate the radiosensitizing effect of NCB-0846 on TNIKhigh LSCC tumors, results showed that pretreatment with NCB-0846 significantly enhanced the efficacy of IR and caused elevated necrosis in TNIKhigh LK2 tumors but not TNIKlow KNS62 tumors.
Mol Cancer Ther. 2024 Apr 27;23(8):1201–11
Mice
Apcmin/þ mice
Oral
22.5-90 mg/kg
BID for 35 days
Suppression of Wnt-driven intestinal tumorigenesis
Nat Commun. 2016 Aug 26;7:12586
SCID mice
A549 cell lung metastasis model
Tail vein injection
3 µM
Single injection, observed for 7 weeks
NCB-0846 significantly reduced the metastatic lesion area in the lungs of SCID mice injected with TGF β1-treated A549 cells.
Br J Cancer. 2021 Jan;124(1):228-236
NOD/SCID mice
Subcutaneous xenograft model
Oral
80 mg/kg
Twice daily for 5 days
Evaluate the inhibitory effect of NCB-0846 on subcutaneous xenografts, results showed significant reduction in tumor volume
搜索
