GW788388

Protein target name:

TGFBR1

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

up to 1 uM, max 10 uM

Probe Availability

MedChemExpress

TOCRIS

Selleck Chemicals

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

TKL

HUGO ID:

HGNC:11772

Entrez gene ID:

7046

UNIPROT ID:

P36897

Target alias:

AAT5; ALK5; ESS1; LDS1; MSSE; SKR4; TBRI; ALK-5; LDS1A; LDS2A; TBR-i; TGFR-1; ACVRLK4; tbetaR-I

Probe Details

Chemical Probes Portal ID:

CPP1667

Probe alias:

GW-788388; GW 788388, cpd 13d

PubChem CID:

10202642

ChEMBL ID:

CHEMBL202887

SMILES string:

C1COCCC1NC(=O)C2=CC=C(C=C2)C3=NC=CC(=C3)C4=C(NN=C4)C5=CC=CC=N5

InChi Key:

SAGZIBJAQGBRQA-UHFFFAOYSA-N

Physico-chemical properties

NMR:

1H NMR (300 MHz, CDCl3) δ 8.58 (m, 2H), 8.40 (d, J = 7.6 Hz, 2H), 8.09 (m, 3H), 7.95 (d, J = 8.1 Hz, 2H), 7.89 (d, J = 7.2 Hz, 1H), 7.41−7.37 (m, 2H), 4.04−4.02 (m, 1H), 3.89 (d, J = 9.4 Hz, 2H), 3.37 (d, J = 11.1 Hz, 2H), 1.77 (d, J = 11.1 Hz, 2H), 1.65−1.58 (m, 2H), NH (pyrazole) not observed;

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

18 nM

Potency assay:

ALK5 Fluorescence Polarization Binding Assay with intracellular kinase domain from ALK5.

Structure-activity relationship:

Yes, see J Med Chem paper

Off target activity

Potency (off target):

IC50

Off target protein and potency:

p38alpha: 7.2 uM

Potency assay (off target):

In an in vitro panel of kinases, GW788388 only inhibited p38a >50% at 10 uM (panel included: P70S6K, AMPK, CHK1, CSK, JNK1, LCK, MKK1, MSK1, MAPK2, PDK1, PhosK, PKA, PKC A, SGK, p38 alpha, p38 beta).

Probe Selectivity in Vitro:

In an in vitro panel of kinases, GW788388 only inhibited p38a >50% (IC50 of 7.28 µM) at 10 uM (panel included: P70S6K, AMPK, CHK1, CSK, JNK1, LCK, MKK1, MSK1, MAPK2, PDK1, PhosK, PKA, PKC A, SGK, p38 alpha, p38 beta). In U2OS cells, GW788388 had no effect on BMP-induced Smad1/5 phosphorylation. Radiometric assay of ALK (caALK)5, TβRII, BMPRII, transfected cells with subsequent IP of the kinase showed activity on TGFBR1, ACVR1B, ACVR1C, TGFBR2; cellular selectivity showed inhibition of ALK4, ALK5 and ALK7 activity.

In cell validation

On target activity in cells

Potency in cells:

IC50

93 nM

Potency assay (cells):

TGF-beta-dependent signaling assay. In 293T cells expressing constitutively active (ca) ALK5, TbetaRII, BMPRII, or ACTRII, 10 uM GW788388 inhibited autophosphorylation of ALK5 and TbetaRII. In U2OS cells expressing caALK1, caALK2, caALK3, caALK4, caALK5, caALK6, or caALK7, GW788388 dose-dependently inhibited only TGF-beta-dependent signaling (ALK4, ALK5, and ALK7; assessed indirectly through Smad-responsive luciferase activity).

Target engagement assay (cells):

In NMuMG (and other cell lines), GW788388 dose-dependently inhibited TGF-beta-induced Smad2 phosphorylation and nuclear translocation; in T47D cells, GW788388 inhibited activin-induced Smad2 phosphorylation.

Off target activity in cells

Potency assay, off target (cells):

In U2OS cells, GW788388 had no effect on BMP-induced Smad1/5 phosphorylation.

In vivo validation

Organism:

Sprague Dawley rat, various mice

Dose:

2 mg/kg, 4 mg/kg

Route of delivery:

OralIntravenous

Plasma half life:

1.3 h (rat, IV)

Systemic clearance:

20 mL/min/kg (rat, IV)

Organ of Interest

Organ (O):

mouse liver, mouse kidney

Target engagement assay:

In DMN-induced mouse models of liver disease and in chemically-induced mouse models of renal fibrosis, GW788388 inhibited TGF-beta-induced collagen IA1 mRNA expression. In db/db mouse model of diabetic retinopathy, GW788388 (2 mg/kg, oral) decreased TGF-beta-induced expression of extracellular matrix (ECM) mRNAs.

Primary Reference

Reference (doi):

10.1038/sj.ki.5002717 10.1021/jm0509905

Reference (PMID):

18075500 16570917

Under Review

This probe is in the process of SAB review. Please continue to check back for new reviews and commentary.

SAB Rating

Rating for use in Cells

2 review(s)

Rating for use in Model Organisms

2 review(s)

SAB Members

SAB Comments

The data for the probe was recorded some time ago. Today, kinase selectivity panels would typically be more comprehensive. Further selectivity testing should be considered.

Aug 26 2021 - 9:45am

The probe validation is a bit meagre and based on one assay from 2006. The assay, a fluorescence polarization displacement assay using rhodamine labelled ATP, is known to have a large variability (much like seen in the original paper) and proven to inflate binding affinities. For a probe to be declared such, I would imagine at the least two more robust and orthogonal validating assays, and perhaps an X-ray structure. Because the rest of the data depends on these considerations, I refrain from commenting on the cellular and in vivo data for now. I wonder if the authors since 2006 have acquired additional binding or inhibition data that confirm the FP assay data reported here.

Sep 21 2021 - 11:35am

GW788388

Probe Availability

Probe Information

Target Details

AAT5; ALK5; ESS1; LDS1; MSSE; SKR4; TBRI; ALK-5; LDS1A; LDS2A; TBR-i; TGFR-1; ACVRLK4; tbetaR-I

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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GW788388

Probe Availability

Probe Information

Target Details

AAT5; ALK5; ESS1; LDS1; MSSE; SKR4; TBRI; ALK-5; LDS1A; LDS2A; TBR-i; TGFR-1; ACVRLK4; tbetaR-I

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?