Brigatinib

Protein target name:

ALK

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

up to 500 nM

Probe Availability

MedChemExpress

Selleck Chemicals

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

RTK

HUGO ID:

HGNC:427

Entrez gene ID:

238

UNIPROT ID:

Q9UM73

Target alias:

Anaplastic lymphoma receptor tyrosine kinase, CD246, NBLST3

Probe Details

Chemical Probes Portal ID:

CPP1202

Probe alias:

AP 26113

PubChem CID:

68165256

ChEMBL ID:

CHEMBL3545311

SMILES string:

CN1CCN(C2CCN(C3=CC=C(NC4=NC=C(Cl)C(NC5=CC=CC=C5P(C)(C)=O)=N4)C(OC)=C3)CC2)CC1

InChi Key:

AILRADAXUVEEIR-UHFFFAOYSA-N

Orthogonal probe:

CeritinibCH5424802TL13-112

Physico-chemical properties

NMR:

1H NMR (400 MHz, CD3OD) δ 8.33 (dd, J = 4.52, 8.03 Hz, 1H), 8.02 (s, 1H), 7.66 (d, J = 8.78 Hz, 1H), 7.59 (ddd, J = 1.51, 7.78, 14.05 Hz, 1H), 7.47–7.54 (m, 1H), 7.25 (ddt, J = 1.00, 2.26, 7.53 Hz, 1H), 6.65 (d, J = 2.51 Hz, 1H), 6.45 (dd, J = 2.51, 8.78 Hz, 1H), 3.84 (s, 3H), 3.69 (d, J = 12.30 Hz, 2H), 2.62–2.86 (m, 6H), 2.43–2.62 (m, 4H), 2.33–2.42 (m, 1H), 2.29 (s, 3H), 1.97–2.08 (m, 2H), 1.83 (d, J = 13.30 Hz, 6H), 1.66 (dq, J = 3.89, 12.09 Hz, 2H).

13C NMR (151 MHz, CDCl3) δ 18.57 (d, J = 71.53 Hz), 28.28 (s), 46.02 (s), 49.01 (s), 50.52 (s), 55.46 (s), 55.65 (s), 61.79 (s), 101.07 (s), 106.01 (s), 108.41 (s), 120.25 (d, J = 95.73 Hz), 120.68 (s), 122.09 (s), 122.41 (d, J = 12.10 Hz), 123.13 (br d, J = 6.60 Hz), 129.48 (d, J = 11.00 Hz), 132.36 (s), 143.91 (d, J = 2.20 Hz), 147.59 (s), 149.38 (s), 154.97 (s), 155.91 (s), 157.82 (s).

31P NMR (162 MHz, CDCl3) δ 43.55.

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

0.6 nM WT, 0.6 nM C1156Y, 1.4 nM F1174L, 1.7 nM L1196M, 4.9 nM G1202R, 6.6 nM R1275Q

Potency assay:

Kinase Assay @ 10 uM ATP

PDB ID for probe-target interaction (3D structure):

6MX8

Structure-activity relationship:

yes

Off target activity

Off target protein and potency:

IGF1R 24.9 nM, InsR 196 nM, FLT3 2.1 nM, ROS1 1.9 nM

Probe Selectivity in Vitro:

In vitro HotSpotSM kinase profiling of 289 kinases: ∼4% (11 out of 289) of the kinases were inhibited with IC50’s of <10 nM. No activity against c-Met or Ron @ 1 uM.

In cell validation

On target activity in cells

Potency in cells:

IC50GI50

29 nM (ALK+), 3194 nM (ALK-)14 nM EML4-ALK, 45 nM EML4-ALK (C1156Y), 55 nM EML4-ALK (F1174L), 41 nM EML4-ALK (L1196M), 194 nM EML4-ALK (G1202R)

Potency assay (cells):

Cell viability assay, CyQuant Cell Proliferation assay

Off target activity in cells

Potency in cells, off target:

IC50

Off target protein and potency (cells):

18 nM CD74-ROS1, 158 nM FLT3-ITD

Probe Selectivity in Cell:

IC50s (nM): >3,000 p-EGFR in EGF-stimulated H358 cells, 397 EGFR (L858R), 489 EGFR (L858R/T790M), 148 p-IGF-1R in IGF1-stimulated HepG2 cells, 9331 p-INSR in insulin-stimulated H-4-II-E cells.

In vivo validation

Organism:

Mouse, Rat

Dose:

3, 10 mg/Kg

Route of delivery:

OralIntravenous

Plasma half life:

4.8 (IV, R), 3.4 (PO, R), 5.8 (PO, M) h

Systemic clearance:

0.46 L/(h·kg) (IV, R)

Cmax:

448 ng/ml (PO, M), 305 ng/ml (PO, R)

Tmax:

4 h (PO, R)

Primary Reference

Reference (doi):

10.1021/acs.jmedchem.6b00306 10.1158/1078-0432.CCR-16-0569

Reference (PMID):

27144831 27780853

Brigatinib

Probe Availability

Probe Information

Target Details

Anaplastic lymphoma receptor tyrosine kinase, CD246, NBLST3

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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Brigatinib

Probe Availability

Probe Information

Target Details

Anaplastic lymphoma receptor tyrosine kinase, CD246, NBLST3

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?