G749

Protein target name:

FLT3

Mechanism of action:

InhibitorATP Competitive

Recommended Concentration for use in cells:

up to 10 nM

Probe Availability

MedChemExpress

Selleck Chemicals

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

HUGO ID:

HGNC:3765

Entrez gene ID:

2322

UNIPROT ID:

P36888

Target alias:

Fms related tyrosine kinase 3 , CD135, FLK-2, FLK2, STK1

Probe Details

Chemical Probes Portal ID:

CPP1210

Probe alias:

G-749, cpd 6

PubChem CID:

78357765

ChEMBL ID:

CHEMBL4544478

SMILES string:

O=C1C2=C(N=C(NC3CCN(C)CC3)N=C2NC4=CC=C(OC5=CC=CC=C5)C=C4)C(Br)=CN1

InChi Key:

SXWMIXPJPNCXQQ-UHFFFAOYSA-N

Orthogonal probe:

AC220

Physico-chemical properties

NMR:

1H NMR (400M Hz, CDCl3) δ (ppm): 11.62 (s, 0.7H), 11.42 (s, 0.3H), 11.09 (br s, 1H), 7.70-7.79 (m, 2H), 7.56 (m, 1H), 7.32-7.39 (m, 2H), 7.02-7.14 (m, 5H), 5.79 (d, J = 7.32Hz, 0.7H), 5.43 (d, J = 7.04Hz, 0.3H), 3.88 (m, 0.3H), 3.75 (m, 0.7H), 2.90 (m, 2H), 2.01-2.37 (m, 7H), 1.66 (m, 2H).

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

0.4 nM FLT3-WT, 0.6 nM FLT3-D835Y

Potency assay:

TR-FRET Kinase Assay

Off target activity

Potency (off target):

IC50

Off target protein and potency:

Mer 1 nM Aurora B 6 nMRET 9 nM VEGFR1/FLT1 18 nMFms 19 nM Axl 20 nM Aurora C 24 nM FGFR1 25 nMFGFR3 30 nMVEGFR2/KDR 39 nMc-Kit 142 nM

Potency assay (off target):

TR-FRET

Probe Selectivity in Vitro:

IGF-1R, PDGFRα, PDGFRβ IC50 > 300 nM

In cell validation

On target activity in cells

Potency in cells:

IC50

WT 6.1 ± 0.4 nM, ITD 11.8 ± 3.7 nM, ITD/N676D 21.4 ± 1.1 nM, ITD/F691L 38.1 ± 6.6 nM, D835Y/N676D 2.1 ± 0.7 nM, D835Y 3.1 ± 0.5 nM

Potency assay (cells):

Cell viability by an ATPLite assay, Western Blot (autophosphorylation, p-ERK, p-AKT) and ELISA assay

In vivo validation

Organism:

Mouse

Dose:

30 mg/Kg

Route of delivery:

Oral

Primary Reference

Reference (doi):

10.1182/blood-2013-04-493916

Reference (PMID):

24532805

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

G749 is a highly potent FLT3 kinase inhibitor that also has good activity against mutations such as D835Y and ITD/F691L that confer resistance to AC220. It has good oral bioavailability. However, its significant activity on other RTK and kinases that are likely to contribute to toxicity limits its use as a mechanistic tool. The compound should be used at a very low concentration (below 10 nM) together with other, more selective FLT3 inhibitors such as AC220 and potentially inhibitors that control for off-target activity in particular Aurora B (IC50 6 nM), MERTK (IC50 1 nM).

Jun 4 2021 - 11:41am

This is a high-quality reagent but should be recognized as a dual Flt3/MerTK inhibitor. However, there are other compounds, such as Gilteritinib, an FDA-approved Flt3 inhibitor with a similar kinome profile (also potent versus MerTK), that might be preferred due to its advanced status. Many (all?) other Flt3 inhibitors also inhibit members of the TAM family of kinases, including MerTK, so although this compound does not meet probe selectivity criteria, it may be a useful tool in cells known to be dependent upon Flt3 ITD for proliferation. Because MerTK becomes ectopically expressed in many leukemias (doi:10.1172/jci.insight.85630), activity in non-Flt3 mutant cell lines is also anticipated.

Jun 18 2021 - 9:51am

The kinase selectivity of this compound is insufficient to make it a good probe, and especially the potent Mer and Aurora B activity are concerning and potentially contribute to the antiproliferative activity of this compound and its in vivo efficacy. The authors of the source paper themselves acknowledge that “the inhibition of aurora B and Mer in addition to inhibition of activated FLT3 disease pathways may contribute to a significant antileukemic effect.” In addition, this compound contains a strongly basic amine, coupled with relatively high lipophilicity, and it would be desirable to see secondary pharmacology profiling data to get a sense of potential promiscuity unrelated to kinase activities. The compound is lacking any pharmacokinetics data to allow for a PK/PD relationship assessment.

Jul 26 2021 - 12:09pm

G749

Probe Availability

Probe Information

Target Details

Fms related tyrosine kinase 3 , CD135, FLK-2, FLK2, STK1

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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G749

Probe Availability

Probe Information

Target Details

Fms related tyrosine kinase 3 , CD135, FLK-2, FLK2, STK1

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?