CFI-402257

Protein target name:

TTK

Mechanism of action:

Type 1.5 Inhibitor, ATP competitive

Recommended Concentration for use in cells:

10-100 nM

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

Other

HUGO ID:

HGNC:12401

Entrez gene ID:

7272

UNIPROT ID:

P33981

Target alias:

CT96, ESK, MPH1, MPS1, MPS1L1, PYT, TTK

Probe Details

Chemical Probes Portal ID:

CPP1683

Probe alias:

cpd 24

PubChem CID:

118086034

ChEMBL ID:

CHEMBL4469414

SMILES string:

Cc1cc(-c2cnn3c(NC[C@H]4C[C@@](C)(O)C4)cc(Oc4cccnc4)nc23)ccc1C(=O)NC1CC1

InChi Key:

PMQUGSPFUBGJCZ-UHFFFAOYSA-N

Orthogonal probe:

BAY1217389EmpesertibS 81694

Physico-chemical properties

NMR:

1H NMR (400 MHz, CD3OD) δ 9.14 (br, s, 1H), 8.89-8.82 (m, 1H), 8.79-8.71(m, 1H), 8.40 (s, 1H), 8.31-8.21 (m, 1H), 7.68 (s, 1H), 7.59 (d, J = 9.5 Hz, 1H), 7.23 (d, J = 8.0 Hz, 1H), 6.06 (s, 1H), 3.56 (d, J = 6.5 Hz, 2H), 2.88-2.79 (m, 1H), 2.40-2.31 (m, 1H), 2.29 (s, 3H), 2.26-2.18 (m, 2H), 1.99-1.89 (m, 2H), 1.37 (s, 3H), 0.85-0.76 (m, 2H), 0.63-0.53 (m, 2H).

13C NMR (100.6 MHz, DMSO-d6) 170.5, 163.4, 150.0, 149.5, 146.5, 144.3, 143.5, 142.0, 135.8, 133.8, 130.4, 127.8, 127.1, 124.7, 121.8, 106.4, 73.7, 68.0, 47.5, 42.0, 28.1, 24.7, 23.2, 20.0, 6.2. H

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

Ki, IC50

0.09 ± 0.02 nM (Ki), 1.7 nM (IC50)

Potency assay:

TTK Inhibition Assay

Structure-activity relationship:

yes

Off target activity

Probe Selectivity in Vitro:

Tested against a panel of human kinases at 1 μM: CFI-402257 inhibited none. The molecule is an intracellular inhibitor of exogenous TTK autophosphorylation with no effect on Aurora kinase A or B or Histone H3 phosphorylation. Outsider target family: CFI-402257 had no activity against CYPs 1A2 and 2D6, inhibited 2C9 and 2C19 with IC50s of 13 and 8 μM, respectively. The level of CYP 3A4 activity reported was substrate dependent with IC50s of 0.51 and 14 μM for BFC and DBF, respectively. CFI-402257 had no activity against CYPs 1A2 and 2D6, inhibited 2C9 and 2C19 with IC50s of 13 and 8 μM, respectively. The level of CYP 3A4 activity reported was substrate dependent with IC50s of 0.51 and 14 μM for BFC and DBF, respectively. Profiled against a panel of 98 nonkinase targets, including G protein-coupled receptors, nuclear receptors, membrane channels, and enzymes @ 10 uM showing only 3 targets with inhibition ≥ 50%.

In cell validation

On target activity in cells

Potency in cells:

GI50, EC50

2 nM (M), 2 nM (H), 4nM (O) (GI50), 6.5 nM

Potency assay (cells):

Cell Viability Assay in MDA-MB468 (M), HCT116 (H), OVCAR-3 (O), Autophosphorylation inhibition assay

In vivo validation

Organism:

Rat, Dog, Mouse

Dose:

1 mg/Kg IV5 mg/Kg PO

Route of delivery:

OralIntravenous

Other route of delivery:

22 (R), 7.9 (D) mL/min/kg

Plasma half life:

2-3 h (M)

Cmax:

1,070 ng/mL (M)

Primary Reference

Reference (doi):

10.1073/pnas.1700234114 10.1021/acsmedchemlett.5b00485

Reference (PMID):

27437075 28270606

Under Review

This probe is in the process of SAB review. Please continue to check back for new reviews and commentary.

SAB Rating

Rating for use in Cells

1 review(s)

Rating for use in Model Organisms

1 review(s)

SAB Members

SAB Comments

This compound appears to be an excellent probe for inhibiting TTK in cells and in mice. Strong structural characterization and biochemical profiling indicate that it is quite selective toward this kinase. The fact that it is highly orally bioavailable and well-tolerated in vivo makes it an attractive probe for this protein. It is now undergoing clinical evaluation in patients with breast cancer.

Aug 5 2021 - 9:05am

CFI-402257

Probe Information

Target Details

CT96, ESK, MPH1, MPS1, MPS1L1, PYT, TTK

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?

Search form

CFI-402257

Probe Information

Target Details

CT96, ESK, MPH1, MPS1, MPS1L1, PYT, TTK

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?