CGI1746

Reagent Authentication Certificate:

Chemical Probes Portal CGI1746.pdf

Protein target name:

BTK

Mechanism of action:

ATP-competitive inhbitor

Recommended Concentration for use in cells:

30 nm - 1 uM

Probe Availability

Cayman (910232-84-7)

MedChem Express (910232-84-7)

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

HUGO ID:

HGNC:1133

Entrez gene ID:

695

UNIPROT ID:

Q06187

Target alias:

AGMX1, AT, ATK, BPK, IMD1, PSCTK1, XLA

Probe Details

Chemical Probes Portal ID:

CPP129

Probe alias:

CGI-1746

PubChem CID:

24857323

ChEMBL ID:

CHEMBL1230541

SMILES string:

CC1=C(C=CC=C1NC(=O)C2=CC=C(C=C2)C(C)(C)C)C3=CN(C(=O)C(=N3)NC4=CC=C(C=C4)C(=O)N5CCOCC5)C

InChi Key:

JIFCFQDXHMUPGP-UHFFFAOYSA-N

Orthogonal probe:

AVL-252GDC-0834

PAINs evaluation:

PAINs free

Physico-chemical properties

NMR:

1H NMR (500 MHz, CDCl3) δ 8.43 (s, 1H), 7.98 (d, 1H, J = 7.5 Hz), 7.87 (m, 4H), 7.78 (s, 1H), 7.54 (dd, 2H, J = 7.0, 1.5 Hz), 7.40 (dd, 2H, J = 7.0, 1.5 Hz), 7.32 (t, 1H, J = 7.5 Hz), 7.22 (d, 1H, J = 7.5 Hz), 6.77 (s, 1H), 3.63 (bs, 8H), 3.60 (s, 3H), 2.38 (s, 3H), 1.37 (s, 9H); 13C NMR (75 MHz, DMSO-d6) δ 169.1, 165.3, 154.4, 150.4, 146.3, 141.4, 138.3, 137.2, 132.5, 131.8, 131.1, 128.5, 128.5, 128.0, 127.5, 127.2, 126.6, 125.5, 125.2, 120.6, 118.5, 66.2, 36.7, 34.7, 31.0, 15.6

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

1.5 nM

Potency assay:

IC50 = 1.9 nM in vitro kinase assays

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

3OCS

Structure-activity relationship:

Hit-to-lead chemistry confirmed the importance of the hydrogen bond donor-acceptor arrangement provided by the amino-imidazopyrazine, identified bioisosteric hinge binding motifs and highlighted the para position of the 8-aniline ring as a promiscuous site tolerant of diverse substitutions. Notably, we found the t-butylphenyl amide to be a critical pharmacophore for potency and selectivity.

Off target activity

Potency assay (off target):

CGI1746 is ~1,000-fold selective over the next kinase in Ambit KinomeScan of 385 kinases and UBI Kinase Profiler of 205 kinases.

Probe Selectivity in Vitro:

Against 82 non-kinase targets, CGI1746 showed <40% inhibition at 1 uM for all targets, and <15% inhibition for 78 of them.

In cell validation

On target activity in cells

Potency in cells:

IC50

~2.9 nM

Potency assay (cells):

Average IC50 for several assay is given. Assays included inhibition of autophosphorylation and protein substrate phosphorylation.

Target engagement assay (cells):

Indirect: inhibition of autophosphorylation and substrate phosphorylation

In vivo validation

Organism:

Balb/c mouse

Dose:

100 mg/kg

Route of delivery:

Subcutaneous

Cmax:

27.2 uM

Tmax:

0.25 h

Primary Reference

Reference (doi):

10.1038/nchembio.481

Reference (PMID):

21113169

CGI1746

Probe Availability

Probe Information

Target Details

AGMX1, AT, ATK, BPK, IMD1, PSCTK1, XLA

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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CGI1746

Probe Availability

Probe Information

Target Details

AGMX1, AT, ATK, BPK, IMD1, PSCTK1, XLA

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?