BMX-IN-1

Reagent Authentication Certificate:

Chemical Probes Portal BMX-IN-1.pdf

Protein target name:

BMX

Mechanism of action:

covalent inhibitor

Recommended Concentration for use in cells:

Up to 25 nM

Probe Availability

MedChem Express (1431525-23-3)

Cayman (1431525-23-3)

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

HUGO ID:

HGNC:1079

Entrez gene ID:

660

UNIPROT ID:

P51813

Target alias:

ETK, PSCTK2, PSCTK3

Probe Details

Chemical Probes Portal ID:

CPP145

Probe alias:

CS-1295, HY-80002, W-6056

PubChem CID:

71576693

SMILES string:

CC1=C(C=C(C=C1)N2C(=O)C=CC3=CN=C4C=CC(=CC4=C32)C5=CC=C(C=C5)NS(=O)(=O)C)NC(=O)C=C

InChi Key:

SFMJNHNUOVADRW-UHFFFAOYSA-N

PAINs evaluation:

PAINs free. Note that no acrylamides were present in the HTS compounds originally used to define PAINS, so "PAINS-FREE" has little meaning here with respect to acrylamides

Physico-chemical properties

NMR:

1H NMR (400 MHz, DMSO-d6)δ9.87 (s, 1 H), 9.78 (s, 1 H), 9.12 (s, 1 H), 8.31 (d, J = 9.4 Hz, 1 H), 8.06 (d, J = 8.5 Hz, 1 H), 8.00 (d, J = 8.5 Hz, 1 H), 7.69 (s, 1 H), 7.53 (d, J = 8.1 Hz, 1 H), 7.24- 7.21 (m, 6 H), 6.92 (d, J = 9.4 Hz, 1 H), 6.58 (dd, J = 17.2 Hz, J = 10.2 Hz,1 H), 6.20 (d, J = 17.2 Hz, 1 H), 5.74 (d, J = 10.2 Hz, 1 H), 3.02 (s, 3 H), 2.42 (s, 3 H). 13C NMR (100 MHz, DMSO-d6)δ163.8, 163.1, 151.3, 148.6, 142.6, 140.5, 138.7, 138.6, 138.4, 136.8, 134.7, 133.2, 132.6, 131.8, 131.2, 128.8, 128.3, 127.7, 125.9, 124.7, 123.2, 122.2, 120.0, 118.0, 113.8, 18.3.

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

8 nM

Potency assay:

Invitrogen SelectScreen Technology

Off target activity

Potency (off target):

IC50

Off target protein and potency:

BTK 10.4 nM

Potency assay (off target):

Assessed activity against 442 kinases using the KinomeScan approach

In cell validation

On target activity in cells

Potency in cells:

IC50

25 nM

Potency assay (cells):

Inhibits the autophosphorylation of BIX.

Target engagement assay (cells):

Indirect: inhibition of autophosphorylation Direct: Immunoprecipitate WT and C496S mutant targets, and BMX-IN-1 inhibits WT protein IC50 = 138 nM and no activity against mutant >10 uM.

Off target activity in cells

Potency in cells, off target:

GI50

Off target protein and potency (cells):

TEL-JAK1 4.92 uMTEL-JAK2 5.83 uMTEL-JAK3 7.98 uMTEL-TYK2 E957D 6.09 uMTEL-BLK 3.64 uM

Potency assay, off target (cells):

Inhibits the proliferation of a panel of murine Ba/F3 cells that were transformed with TEL fusion of BMX.

Primary Reference

Reference (doi):

10.1021/cb4000629

Reference (PMID):

23594111

Endorsed probe

The Chemical Probes Portal endorses this compound as chemical probe for use in cells.

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

no rating

SAB Members

SAB Comments

Based on the data in Supplementary Table 1, this probe also binds to 63 other kinases (score less than or equal to 75%) at 1 micromolar concentration. So choosing a concentration well below 1 micromolar is critical for cell based studies. In addition, this probe is equipotent against BTK. BMX-IN-1 shows better selectivity than other known inhibitors of BMX, but even this probe can bind (and perhaps inhibit) a number of other kinases at higher concentrations.

Jun 30 2016 - 12:30am

BMX-IN-1 is an irreversible inhibitor of the kinase BMX and is the best available probe for it. It is also a potent inhibitor of BTK. In a broad kinome scan experiment (DiscoverX technology), BMX-IN-1 inhibits only 1% of the 442 kinases tested at >90% at a concentration of 1 micromolar. This is excellent selectivity. That said, there are potential off targets, so care should be taken in cell-based experiments to keep the dose as low as possible. The on-target, cell-based potency in a TEL-BMX transformed Ba/F3 cell line system is 25 nM. 1 micromolar of BMX-IN-1 inhibits autophosphorylation of BMX in RV-1 cells.

Sep 12 2016 - 6:29pm

BMX-IN-1

Probe Availability

Probe Information

Target Details

ETK, PSCTK2, PSCTK3

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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BMX-IN-1

Probe Availability

Probe Information

Target Details

ETK, PSCTK2, PSCTK3

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?