The TGF-β pathway regulates a wide variety of cellular processes in many different cell types and biological contexts. There are three identified TGF receptor ligands, TGF-β1, 2, and 3. Activated TGF-β ligands can interact with TGF-β type II receptors (TβRII), then recruit and phosphorylate the TGF-β type I receptors (also called as TβRI or ALK5). In turn, activated TβRI phosphorylates SMAD2 and SMAD3 at C-terminal serine residues. Following that, phosphorylated SMAD and SMAD3 assemble into heterodimeric and trimeric complexes with SMAD4. Then the trimeric complex translocate into the nucleus and regulate the expression of TGF-β target genes[1]. SB-525334 is an ALK5 inhibitor with IC50 value of 14.3 nM, with modest potent to ALK4 with an IC50 value of 58.5 nM (measured by GST-ALKs kinase assay). Addition of 1 μM SB-525334 reduced TGF-β1-induced Smad2 and Smad3 nuclear localization back to the control levels in RPTE cells[2]. SB-525334 on concentration of 0.5 - 2 μM can inhibit p-SMAD2 in dose-dependent manner in ELT-3 cells[3]. SB525334, 30 mg/kg, given twice daily by oral gavage for 2 weeks can attenuate f bleomycin-induced lung fibrosis in mice[4].
SB-525334 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Healthy NK cells
2 µM
1-hour pre-incubation
Prevented POD1 plasma-induced NK cell dysfunction, maintained NKG2D, DNAM-1, CD132 expression and IFNγ production
Int J Mol Sci. 2022 Nov 23;23(23):14608
Rabbit bone marrow mesenchymal stem cells (R-BMSCs)
14.3 nM
4 hours
To evaluate the effect of SB-525334 on the survival rate of R-BMSCs, results showed that SB-525334 had no significant effect on the survival rate of R-BMSCs under 48˚C microwave treatment.
Mol Med Rep. 2020 Aug;22(2):906-914
VX2 cells
14.3 nM
4 hours
To evaluate the effect of SB-525334 on the survival rate of VX2 cells, results showed that SB-525334 significantly reduced the survival rate of VX2 cells under 48˚C microwave treatment.
Mol Med Rep. 2020 Aug;22(2):906-914
Immortalized human hepatic stellate cells (hTERT-HSC)
1 µM
48 hours
SB52 inhibits TGF-β1-induced activation of hTERT-HSC.
PLoS One. 2018 Jul 20;13(7):e0201044
Human primary hepatic stellate cells
1 µM
48 hours
SB52 induces upregulation of Nrf2 and Nqo1, efficiently inhibiting the TGF-β1 effect.
PLoS One. 2018 Jul 20;13(7):e0201044
PASMCs from familial iPAH patients
1 µM
6 days
To evaluate the effect of SB525334 on TGF-β1-mediated proliferation of PASMCs from familial iPAH patients. Results showed that SB525334 significantly inhibited TGF-β1-mediated proliferation.
Am J Pathol. 2009 Feb;174(2):380-9
Lgr5+CD44+EpCAM+ colorectal cancer stem cells
2 μg/mL
6 hours
SB525334 inhibited the TGF-β1 pathway, downregulated TGF-β1, p-Smad2/3, and Snail expression, while upregulated Smad4, E-cad, and ZO-1 expression, thereby suppressing KLF4 overexpression-induced mesenchymal phenotypes and stemness properties.
J Cell Mol Med. 2020 Jan;24(2):1866-1877
Human umbilical vein endothelial cells (HUVECs)
10 µM
72 hours
To investigate the role of TGF-β receptor ALK5 signaling in HUVEC morphology under shear stress. Results showed that SB525334 inhibition of ALK5 signaling led to disruption of the HUVEC monolayer and increased apoptosis.
SB 525334 treatment blocked TGF-β-induced upregulation of ADAM10, ADAM17, ADAM12, and ADAM19 gene expression.
Am J Pathol. 2013 Dec;183(6):1885-1896
SU-DIPG-IV cells
50 µM
72 hours
Evaluate the inhibitory effect of SB525334 on SU-DIPG-IV cell growth, showing significant inhibition at 50 µM dose
Sci Rep. 2020 Apr 9;10(1):6140
SF8628 cells
50 µM
72 hours
Evaluate the inhibitory effect of SB525334 on SF8628 cell growth, showing significant inhibition at 50 µM dose
Sci Rep. 2020 Apr 9;10(1):6140
CD44+ leader cells
5 µM
from day 0 thoursoughours day 3 post-injury
To investigate the role of endogenous TGF β signaling in CD44+ leader cells during wound healing. Results showed that endogenous TGF β signaling had no effect on leader cell-directed wound closure.
Exp Eye Res. 2021 Dec;213:108829
SB-525334 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Sprague-Dawley rats
High-salt diet-induced endothelial dysfunction model
Oral (via drinking water)
10 mg/kg/day
Once daily for 2 days
SB525334 reduced high-salt diet-induced Smad2 phosphorylation to levels observed in rats on the low-salt diet and prevented the downstream signaling events induced by the high-salt diet
Hypertension. 2013 Nov;62(5):951-6
C57/BL6 mice
Unilateral ureteral obstruction (UUO) model
Oral gavage
10 mg/kg/day
Once daily for 7 days
SB 525334 significantly reduced the gene expression of ADAM10, ADAM17, ADAM12, and ADAM19 in the UUO model and decreased the enzymatic activity of ADAM10 and ADAM17.
Am J Pathol. 2013 Dec;183(6):1885-1896
New Zealand rabbits
Osteosarcoma model
Ear vein injection
21.45 nM, 100 µL
21 days
To evaluate the therapeutic effect of SB-525334 on osteosarcoma, results showed that microwave treatment combined with SB-525334 significantly reduced tumor volume.
Mol Med Rep. 2020 Aug;22(2):906-914
Sprague-Dawley rats
MCT-induced pulmonary arterial hypertension model
Oral
3 or 30 mg/kg
Daily dosing until day 35
To evaluate the therapeutic effect of SB525334 on MCT-induced pulmonary arterial hypertension. Results showed that SB525334 significantly reversed elevated pulmonary arterial pressure and inhibited right ventricular hypertrophy.
Am J Pathol. 2009 Feb;174(2):380-9
Chick embryos
Ex vivo mock-cataract-surgery model
In vitro culture treatment
5μM
From day 0 through day 3 post-injury
To investigate the role of endogenous TGF β signaling in the fibrotic response of CD44+ leader cells in the rigid wound environment. Results showed that endogenous TGF β signaling induced FN-EDA and pro-collagen I production by leader cells and promoted their transition to an αSMA+ myofibroblast phenotype in the rigid environment.
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