CH5424802

Reagent Authentication Certificate:

Chemical Probes Portal CH5424802.pdf

Protein target name:

ALK

Mechanism of action:

ATP-competitive inhibitor

Recommended Concentration for use in cells:

Up to 50 nM

Probe Availability

MedChem Express (1256580-46-7)

Cayman (1256580-46-7)

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:

CD246, NBLST3

Probe Details

Chemical Probes Portal ID:

CPP121

Probe alias:

Alectinib

PubChem CID:

49806720

ChEMBL ID:

CHEMBL1738797

SMILES string:

CCC1=C(C=C2C(=C1)C(=O)C3=C(C2(C)C)NC4=C3C=CC(=C4)C#N)N5CCC(CC5)N6CCOCC6

InChi Key:

KDGFLJKFZUIJMX-UHFFFAOYSA-N

Orthogonal probe:

CeritinibBrigatinibTL13-112

PAINs evaluation:

PAINs free

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

0.83 nM

Potency assay:

In vitro kinase assay: ALK L1196M IC50 = 1.56 nM; ALK IC50 = 1.9 nM, KD = 2.4 nM (Ambit)

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

3AOX

Off target activity

Potency assay (off target):

CH5424802 was tested against 402 kinases with >50% inhibition at 10 nM of ALK, GAK, and LTK only.

In cell validation

On target activity in cells

Potency assay (cells):

In cells, CH5424802 dose-dependently inhibited ALK and ALK L1196M autophosphorylation and protein substrate phosphorylation.

Target engagement assay (cells):

Indirect, inhibition of protein substrate phosphorylation

In vivo validation

Organism:

mice

Dose:

6 mg/kg

Route of delivery:

Oral

Plasma half life:

8.6 hr

Cmax:

1707 nM

Tmax:

2 hr

Organ of Interest

Organ (O):

xenograft

Target engagement assay:

Indirect: inhibition of ALK autophosphorylation

Primary Reference

Reference (doi):

10.1016/j.ccr.2011.04.004

Reference (PMID):

21575866

Endorsed probe

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

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

CH5424802 (alectinib) is a second generation ALK inhibitor. The compound inhibits the ALK gate-keeper mutation L1196M, one of the commonly seen secondary mutations leading to resistance to crizotibib, with an IC50 of 1.5 nM.

Jun 12 2016 - 3:19am

This molecule is a potent, selective and orally bioavailable. It is highly selective (1.9 nM biochemical assay; 2.4 nM Kd) against target, with only 2 other kinases showing >50% inhibition at 10 uM (out of a panel of 402). Extensive cell-based data, target engagement, and downstream-marker level changes have been recorded. This molecule has been designed for greater selectivity and to overcome resistance mutations. As such, care should be taken when using as probe in checking the relevance of the mutated ALK form being studied. The original publication contains convincing data including protein-drug co-crystal structure. PK data included in mice: half-life (8.6 hr), oral bioavailability (70.8%). ED50 = 0.46 mg/kg in NCI-H2228 model using once daily, oral delivery.

Aug 17 2016 - 7:43pm

This molecule is recommended for isolated enzyme, cell, 3D-cell spheroid and animal work for the specific ALK mutant described (ALK L1196M). There is a convincing package of on- and off-target selectivity, cell, biophysical and mouse data. Multiple lines of evidence demonstrate direct inhibition of the enzyme (biochemical, competition experiments, biophysical and X-ray) and exceptional selectivity for this ATP-competitive kinase inhibitor (only 3 kinases - ALK, GAK, LTK are inhibited >50% at 10 nM across a 402 kinase panel). Extended exposure results from a single dose in mice (high 70% oral bioavailability with 8.6 h half life).

Sep 8 2016 - 3:56pm

This is an ATP-competitive inhibitor of ALK kinase with IC50=1.9nM and KD=2.4 nM. It is active on the wild-type protein and the L1196M gatekeeper mutant that confers resistance to other inhibitors. The compound binds with one H-bond (crystal structure) to the kinase hinge region (as opposed to two or three found with other inhibitors), which might explain its selectivity. The inhibitor was selective in a small panel of 24 kinases and in the Ambit panel with 402 kinases. Only GAK and TLK were inhibited as well (window of ca. 5). The compound is active in NSCLC- and other tumor cell lines expressing the EML4-ALK fusion oncogene but not in ALK-fusion-negative cells. The inhibitor induces tumor regression in mouse tumor cell xenografts expressing native EML4-ALK or the L1196M getekeeper mutation.

Jan 2 2017 - 3:16pm

CH5424802

Probe Availability

Probe Information

Target Details

CD246, NBLST3

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?

Search form

CH5424802

Probe Availability

Probe Information

Target Details

CD246, NBLST3

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?