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[3]. SB-431542 is a selective ALK5 inhibitor with IC50 value of 94nM (measured by GST-ALK5 kinase activity)[1]. SB-431542 also shows inhibition against ALK4 and ALK7, which are very similar to ALK5 in their kinase domains. Addition of SB-431542 for 24h at a concentration of 10uM can efficiently inhibit the p-Smad2 phosphorylated by activated ALK4, ALK5, and ALK7 in NIH 3T3 cells, with no effect on BMP induced phosphorylation of Smad1, which is mediated by ALKs 2, 3, and 6. The transcription activity mediated by the activated endogenous ALKs 4, 5 and 7 can also inhibited dose-dependently by 0.25, 0.5, 0.75, 1, 2, 5, or 10uM SB-431542[2]. SB-431542 has performance in both reprogramming and differentiation, as 1. SB-431542 can replace SOX2 in the reprogramming of mouse fibroblasts to iPSCs; 2. Combined with PD0325901 and Thiazovivin, SB-431542 can increase the efficiency of reprogramming human somatic cells to iPSCs; 3. SB-431542, combined with CHIR99021, ISX-9, Forskolin and I-BET151, can direct lineage reprogramming of fibroblasts to mature neurons; 4. SB-431542, in combination with either LDN193189 or Noggin, promotes the differentiation of NPCs from human PSCs; 5. SB-431542 can promote the proliferation and sheet formation of mouse ES-derived endothelial cells; 6. SB-431542 can enhance differentiation of cardiomyocytes.
作用机制
SB-431542 acts as a competitive ATP binding site kinase inhibitor[2].
SB 431542 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Human cord blood (CB)-derived CD34+ hematopoietic stem and progenitor cells (HSPCs)
5 µM
15 days
To evaluate the ability of SD208 and SB431542 to increase erythropoiesis in human DBA models, results showed that SD208, SB431542, and Galunisertib significantly increased erythroid expansion in RPS19-insufficiency.
Nat Commun. 2020 Jul 3;11(1):3344.
Lin-Kit+ progenitors from mouse embryonic (E14.5–E15.5) fetal livers
10 µM
4 days
To screen for small molecules that increase erythroid expansion in mouse models of DBA, SB431542 and SD208 significantly rescued c-Kit+ erythroid expansion.
Nat Commun. 2020 Jul 3;11(1):3344.
camel granulosa cells
20 µM
20 h
Inhibited TGFβ pathway and maintained spindle morphology of cells under chronic heat stress
J Adv Res. 2019 Nov 22;22:105-118.
Tendon-derived stem cells (TDSCs)
10 µM
72 h
To explore the role of TGF-β1 and mTOR signaling in HUMSC-Exos-mediated tendon regeneration. Results showed that SB-431542 inhibited the promoting effects of HUMSC-Exos on tendon marker expression.
J Nanobiotechnology. 2021 Jun 5;19(1):169.
SB 431542 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
BALB/c mice
4T1 mouse mammary tumor model
Intratumoral injection
50 mg/kg
Single injection, observed for 100 days
Evaluate the therapeutic effect of rGO/MTX/SB under laser irradiation, showing significant inhibition of tumor recurrence and prolonged survival time
Inhibits Trypanosoma cruzi Y infection-induced TGFbeta signaling in mink C32 cells at 10 uM
17526757
CHO-HIR
0.01-3 μM
Function Assay
2 h
Inhibits TGFbeta-induced downstream transcriptional activation of ALK5 expressed in CHO-HIR cells assessed as intracellular translocation of EGFP-Smad2 with IC50 of 0.35μM
24055046
H1299
1 μM
Migration Assay
12-24 h
Induces antimigratory activity against human H1299 cells assessed as Inhibition of cell migration with IC50 of 0.5μM
24417479
HaCaT
3.2-50 μM
Function Assay
15 min
Inhibits TGFbeta receptor in human HaCaT cells assessed as Smad phosphorylation with IC50 of 0.172μM
20919678
HaCaT
0.05 μM
Function Assay
2 h
Does not inhibit TGF-beta induced ALK5 activity in HaCaT cells assessed as p3TP-luciferase reporter activity at 0.05 uM
17552507
HEK293T
10 μM
Function Assay
22 h
Inhibits TGBR2 signaling in human HEK293T cells assessed as Inhibition of SMAD activation with IC50 of 0.066μM
23130626
HepG2
Function Assay
12 h
Inhibits TGFR-1 in human HepG2 cells expressing PAI-luciferase with IC50 of 0.25μM
19914068
Mouse cardiomyocytes
10 μM
Antimicrobial Assay
4 h
Induces antitrypanosomal activity against Trypanosoma cruzi Y in mouse cardiomyocytes assessed as reduction of intracellular amastigotes at 10 uM
17526757
Mouse cardiomyocytes
10 μM
Antimicrobial Assay
96 h
Induces antitrypanosomal activity against Trypanosoma cruzi Y in mouse cardiomyocytes assessed as Inhibition of trypomastigote release at 10 uM
17526757
Mouse embryo cardiomyocytes
10 μM
Function Assay
1 h
Inhibits invasion of Trypanosoma cruzi Y in mouse embryo cardiomyocytes assessed as pathogen infection at 10 uM
17526757
Mouse embryo cardiomyocytes
10 μM
Function Assay
1 h
Inhibits TGF-beta-1-induced Smad2 phosphorylation in mouse embryo cardiomyocytes at 10 uM
17526757
Mouse embryo cardiomyocytes
10 μM
Function Assay
1 h
Inhibits Trypanosoma cruzi Dm28C infection-induced Smad2 phosphorylation in mouse embryo cardiomyocytes at 10 uM
17526757
Sf9
Function Assay
2 h
Inhibits human recombinant ALK5 phosphorylation expressed in Sf9 cells with IC50 of 1.542μM
17552507
Trypanosoma cruzi trypomastigotes
10 μM
Antimicrobial Assay
4 h
Induces antitrypanosomal activity against Trypanosoma cruzi trypomastigotes assessed as effect on parasite morphology at 10 uM
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