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SB431542

Name: SB431542
Brand: Selleck Chemicals
SKU: S1067
Rating: 5 (657 reviews)

Catalog No.S1067 Purity: 99.99%

SB431542 is a potent and selective inhibitor of ALK5 with IC50 of 94 nM in a cell-free assay, 100-fold more selective for ALK5 than p38 MAPK and other kinases.

SB431542 Chemical Structure

CAS: 301836-41-9

Selleck's SB431542 has been cited by 657 publications

Purity & Quality Control

Batch: Purity: 99.99%

99.99

Products often used together with SB431542

LDN-193189

SB431542 and LDN-193189, along with other small molecules, reduce the expression of epithelial-mesenchymal cell transition (EMT) marker genes and effectively inhibit fibrosis of liver cells.

Xiang C, et al. Science. 2019 Apr 26;364(6438):399-402.

Y-27632 2HCl

SB431542 and Y-27632 2HCl combination reverse the mesenchymal actin cytoskeleton stress fibers induced by TGF-β1 in TGF-β1 knockout mice cells (mTEC-KO).

Das S, et al. BMC Cell Biol. 2009 Dec 21;10:94.

DMH1

SB431542 and DMH1 combined use regulates the two TGF-β signaling pathways independently in human-induced pluripotent stem cells (hiPSCs).

Neely MD, et al. ACS Chem Neurosci. 2012 Jun 20;3(6):482-91.

SU5402

SB431542 and SU5402 block NODAL and FGF9 signalling that allows germ cells to differentiate in Emx2-knockdown testes in vitro.

Tian-Zhong M, et al. Reproduction. 2016 Jun;151(6):673-81.

Laduviglusib (CHIR-99021)

SB431542 and Laduviglusib suppress the gene expressions of pluripotent-related transcription factors and epithelial‑mesenchymal transition (EMT)-related genes to various levels in HeLa cells.

Zhang Y, et al. Oncol Rep. 2019;41(6):3545-3554.

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Cell Data

Cell Lines	Assay Type	Concentration	Incubation Time	Formulation	Activity Description	PMID
HEK293T	Function Assay	10 μM	22 h	DMSO	Inhibits TGBR2 signaling in human HEK293T cells assessed as Inhibition of SMAD activation with IC50 of 0.066μM	23130626
H1299	Migration Assay	1 μM	12-24 h	DMSO	Induces antimigratory activity against human H1299 cells assessed as Inhibition of cell migration with IC50 of 0.5μM	24417479
HaCaT	Function Assay	3.2-50 μM	15 min	DMSO	Inhibits TGFbeta receptor in human HaCaT cells assessed as Smad phosphorylation with IC50 of 0.172μM	20919678
HepG2	Function Assay		12 h	DMSO	Inhibits TGFR-1 in human HepG2 cells expressing PAI-luciferase with IC50 of 0.25μM	19914068
CHO-HIR	Function Assay	0.01-3 μM	2 h	DMSO	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
Sf9	Function Assay		2 h	DMSO	Inhibits human recombinant ALK5 phosphorylation expressed in Sf9 cells with IC50 of 1.542μM	17552507
C32	Function Assay	10 μM	20h		Inhibits Trypanosoma cruzi Y infection-induced TGFbeta signaling in mink C32 cells at 10 uM	17526757
Mouse embryo cardiomyocytes	Function Assay	10 μM	1 h		Inhibits invasion of Trypanosoma cruzi Y in mouse embryo cardiomyocytes assessed as pathogen infection at 10 uM	17526757
Mouse embryo cardiomyocytes	Function Assay	10 μM	1 h		Inhibits TGF-beta-1-induced Smad2 phosphorylation in mouse embryo cardiomyocytes at 10 uM	17526757
Mouse embryo cardiomyocytes	Function Assay	10 μM	1 h		Inhibits Trypanosoma cruzi Dm28C infection-induced Smad2 phosphorylation in mouse embryo cardiomyocytes at 10 uM	17526757
Trypanosoma cruzi trypomastigotes	Antimicrobial Assay	10 μM	4 h		Induces antitrypanosomal activity against Trypanosoma cruzi trypomastigotes assessed as effect on parasite morphology at 10 uM	17526757
Mouse cardiomyocytes	Antimicrobial Assay	10 μM	4 h		Induces antitrypanosomal activity against Trypanosoma cruzi Y in mouse cardiomyocytes assessed as reduction of intracellular amastigotes at 10 uM	17526757
Mouse cardiomyocytes	Antimicrobial Assay	10 μM	96 h		Induces antitrypanosomal activity against Trypanosoma cruzi Y in mouse cardiomyocytes assessed as Inhibition of trypomastigote release at 10 uM	17526757
HaCaT	Function Assay	0.05 μM	2 h	DMSO	Does not inhibit TGF-beta induced ALK5 activity in HaCaT cells assessed as p3TP-luciferase reporter activity at 0.05 uM	17552507
Click to View More Cell Line Experimental Data

Biological Activity

Description

SB431542 is a potent and selective inhibitor of ALK5 with IC50 of 94 nM in a cell-free assay, 100-fold more selective for ALK5 than p38 MAPK and other kinases.

Targets

ALK4 ^[2] (Cell-free assay)	ALK7 ^[2] (Cell-free assay)	ALK5 ^[1] (Cell-free assay)
		94 nM

In vitro
In vitro	SB 431542 inhibits the activin type I receptor ALK4 and the nodal type I receptor ALK7, which are responsible for the phosphorylation of Smad2. SB 431542 has little effect on ALK1, ALK2, ALK3, and ALK6, which show phosphorylation of Smad1. SB 431542 is a selective inhibitor of endogenous activin but has no apparent effect on BMP signaling. SB 431542 could induce both Smad2/Smad4- and Smad3/Smad4-dependent transcription. ^[2] In A498 cells, SB 431542 inhibits both TGF-β1-induced collagen Iα1 and PAI-1 mRNA with IC50 of 60 nM and 50 nM, respectively. In addition, SB 431542 inhibits production of TGF-β1-induced fibronectin mRNA and protein with IC50 of 62 nM and 22 nM, respectively. ^[3] SB 431542 blocks the TGF-β-mediated growth factors, including PDGF-A, FGF-2 and HB-EGF, leading to an increase in proliferation of MG63 cells. SB 431542 also inhibits TGF-β-induced c-Myc and p21 ^WAF1/CIP1. ^[4] SB 431542 significantly suppresses TGF-β-induced G1 arrest, leading to accumulation of cells in the S phase of the cell cycle in FET, RIE, and Mv1Lu cells. SB 431542 also inhibits TGF-β-induced epithelial to mesenchymal transition (EMT) in NMuMG and PANC-1 cells. ^[5] SB 431542 significantly elevates the expression of CD86 in BM-DCs and that of CD83 within CD11c+ cells suppressed by TGF-β. SB 431542 is able to induce NK activity through functional maturation and IL-12 production of human DCs. ^[6]
Kinase Assay	Flashplate assay for ALK5
Kinase Assay	SB 431542 is dissolved in DMSO at a concentration of 10 mM. The kinase domain of TGFβRI, from amino acid 200 to the C-terminus, and the full-length Smad3 protein are expressed as N-terminal glutathion S-transferase (GST) fusion proteins in the baculovirus expression system. Proteins are purified with glutathion Sepharose beads 4B. Basic FlashPlates are coated with 0.1 M sterile filtered sodium bicarbonate, pH 7.6, containing 700 ng of GST-Smad3 per 100 μL. Assay buffer contains 50 mM HEPES (pH 7.4), 5 mM MgCl₂, 1 mM CaCl₂, 1 mM DTT, 100 mM GTP, 3 μM ATP plus 0.5 μCi/well ɤ³³P-ATP, and 85 ng of GST-ALK5 with or without SB 431542. Plates are incubated at 30 °C for 3 hours. The assay buffer is removed by aspiration, and the plate is counted on a Packard TopCount 96-well scintillation plate reader.
Cell Research	Cell lines	MG63 and NIH3T3
	Concentrations	0.3 μM
	Incubation Time	30 minutes
	Method	To explore the effects of ligands, MG63 and NIH3T3 cells are seeded at a density of 8 × 10⁴ cells/well in 6-well plates and starved (0.1% FCS for MG63 cells and 0.5% FCS for NIH3T3 cells) for 24 hours before ligand stimulation. Media containing various ligands are exchanged at 48-hours intervals. Cells are trypsinized and counted by a Coulter counter on days 2, 4, and 6 after ligand stimulation. To explore the effects of constitutively active receptors, cells are seeded at a density of 2 × 10⁵ cells/well in 6-well plates. The next day, cells are infected with adenoviruses carrying various cDNAs at a multiplicity of infection of 100. Cells are trypsinized and counted on day 3. Cell proliferation assay is performed in the presence of 0.3 μM SB 431542.
Experimental Result Images	Methods	Biomarkers	Images	PMID
	Western blot	phospho-SMAD3 / Phospho-p38 Y397 phospho-FAK pSmad2 / p-AKT / E-cadherin / vimentin / snail / slug CK18 / Fibronectin / Vimentin		17113264
	Growth inhibition assay	Cell viability		28125630
	Immunofluorescence	Smad2/3		19619490
	Immunofluorescence	Na+/K+-ATPase		23451286
	ELISA	collagen 1		23451286

In Vivo
In vivo	SB 431542 triggers cytotoxic T lymphocyte (CTL) activities in the colon-26 carcinoma models and is most likely to produce antitumor immunological outcomes through alteration of DC function suppressed by TGF-β. ^[6]
Animal Research	Animal Models	BALB/c mice receive intraperitoneal (i.p.) injections of colon-26 tumor cells.
	Dosages	1 μM solution, 100 μL/mouse
	Administration	Directly injected into peritoneal cavity

References

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Chemical lnformation & Solubility

Molecular Weight	384.39	Formula	C₂₂H₁₆N₄O₃
CAS No.	301836-41-9	SDF	Download SB431542 SDF
Smiles	C1OC2=C(O1)C=C(C=C2)C3=C(NC(=N3)C4=CC=C(C=C4)C(=O)N)C5=CC=CC=N5
Storage (From the date of receipt)	3 years-20°Cpowder	Storage of Stock Solutions Aliquot the stock solutions to avoid repeated freeze-thaw cycles	1 year-80°Cin solvent
Storage (From the date of receipt)	3 years-20°Cpowder		1 month-20°Cin solvent

In vitro
Batch:

DMSO : 77 mg/mL ( (200.31 mM)； Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 3 mg/mL

Water : Insoluble

Molecular Weight Calculator

In vivo Batch: Add solvents to the product individually and in order.					In vivo Formulation Calculator
	Clear solution	0.1%DMSO 1 99.80% Saline	0.02mg/ml (0.05mM)	Taking the 1 mL working solution as an example, add 1 μL of 20 mg/ml clear DMSO stock solution to 998 μL of Saline and mix evenly. The mixed solution should be used immediately for optimal results.	In vivo Formulation Calculator
Clear solution	5%DMSO 1 95% Corn oil	4.0mg/ml (10.41mM)	Taking the 1 mL working solution as an example, add 50 μL of 80 mg/mL clarified DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.
Clear solution	5%DMSO 1 40%PEG 300 2 5%Tween80 3 50% ddH2O	4.0mg/ml (10.41mM)	Taking the 1 mL working solution as an example, add 50 μL of 80 mg/mL clarified DMSO stock solution to 400 μL PEG300, mix evenly to clarify it; add 50 μL Tween-80 to the above system, mix evenly to clarify it; then Continue to add 500 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.

Preparing Stock Solutions

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
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Dilution Calculator Molecular Weight Calculator

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg Average weight of animals: g Dosing volume per animal:μL Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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Frequently Asked Questions

Question 1:
I would appreciate it if you can help me in figuring out the formulation for this drug in vivo experiments.

Answer:
S1067 SB431542 in 1% DMSO+30% polyethylene glycol+1% Tween 80 at 30 mg/ml is a suspension for oral gavage. It can also be dissolved in 2% DMSO+30% PEG 300+ddH2O at 5 mg/ml as a clear solution for IP injection.

TGF-beta/Smad Signaling Pathway Map

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