CCT251545 ATP-competitive inhibitor of CDK19, CDK8



Protein target names: CDK19, CDK8

Mechanism of action: ATP-competitive inhibitor

Recommended in-cell concentration:
35-350 nM

In Vitro Validations

Uniprot ID: Q9BWU1
Target Class: Kinase
Target SubClass: CMGC
Potency: IC 50
Potency Value: 6 nM
Potency Assay: Reporter Displacement Assay for CDK8 and CDK19 provided by Proteros Biostructures GmbH. The assay is based upon the competitive displacement of a reporter probe designed to selectively target the ATP binding site of CDK8 or CDK19 with a fast binding kinetic signature. Binding of the probe to its target results in the emission of an optical signal. Competitive displacement of the probe by the corresponding compounds results in a loss of the optical signal that can be quantified with increasing compound concentrations. Methods Enzymol. 493, 299–320 (2011)
PDB ID for probe-target interaction (3D structure): 5BNJ
Structure-activity relationship: Structure-activity relationship is available in J. Med. Chem. 2015, 58, 1717-1735, Nat. Chem. Biol. 2015, 11, 973-980 and J. Med. Chem. 2016, 59, 1078-1101. The SAR was determined using the 7dF3, LS174T reporter assays and a CDK8 biochemical assay.
Target aliases:
Cyclin-dependent kinase 19, KIAA1028, CDK11, CDC2L ...

DOI Reference: 10.1038/nchembio.1952

In Cell Validations

In Vivo Data

Reagent authentication certificate: PDF image

I have extra information to add

SERP ratings and comments

SERP Ratings

In Cell Rating
In Model Organisms

(last updated: 10 Mar 2017 )

SERP Ratings

In Cell Rating
In Model Organisms

SERP Comments:

CCT251545 was optimized from a phenotypic screening hit for inhibitors of WNT pathway activity. Impressively, this optimization was done without the benefit of structure-based design. CCT251545 has potent cell-based activity and good oral pharmokinetics (PK). The cellular targets of CCT251545 were subsequently determined to be CDK8/19 using SILAC-based affinity capture and rationalized by X-ray crystallography. The affinity-capture experiment suggests very high specificity for CDK8/19. However, direct kinase selectivity profiling experiments did not include the positive controls of CDK8 or CDK19. Therefore, a specificity score cannot be determined nor can it be shown that the compound was active in the assay. Efforts to connect CDK8/19 binding to cellular activity (e.g., WNT reporter gene-assay inhibition) would benefit from the use of a dominant drug resistant allele of CDK8/19. Such an allele could be expressed in cells to circumvent CDK8/19 inhibitory activity of CCT251545 and provide strong evidence that modulation of CDK8/19 by CCT251545 accounts for the observed cellular activity. Regardless, CCT251545 is a potent inhibitor of CDK8/19, has good PK properties and is expected to be highly specific for CDK8/19.

(last updated: 15 Mar 2017 )

SERP Ratings

In Cell Rating
In Model Organisms

SERP Comments:

CCT251545 is a high-affinity ligand of CDK8 and CDK19 with biochemical IC50s for these targets of 7 and 6 nM, respectively. The compound exhibits high selectivity compared with other targets, as demonstrated by proteomics data (SILAC), and suggested by testing in an expanded kinase panel. CCT251545 exhibits potent target engagement in cells (reduction of phospho-STAT1-Ser727; IC50=9 nM) at concentrations consistent with those that reduced WNT-reporter activity in the same cell lines. In vivo pharmacokinetic properties were investigated in mice, rats, and dogs and found adequate for in vivo testing (moderate clearance and F >50% at a low oral dose of 0.5 mg/kg). CCT25145 inhibits the growth of MMTV-WNT-1 breast cancers transplanted from genetically engineered mice as well as APC-mutant SW620 human colorectal cancer xenografts, consistent with in vivo pharmacodynamic data (reduction of p-STATSer727) and at exposure levels that exceed cellular GI50's. Taken together, CCT251545 is a well suited and selective probe for CDK8/19 with proven cellular and in vivo target engagement and efficacy.

(last updated: 18 Mar 2017 )

Portal Comments

CCT251545 and an analogue have been independently profiled using a panel of cell-permeable energy transfer probes to quantify target occupancy for all 21 human CDKs in live cells (Wells et al). They reported that the live-cell occupancy results for CDK8 and CDK19 (≤10 nM Kd-apparent for both CDK8 and CDK19) agreed closely with the single-digit nanomolar cellular potencies reported previously using downstream CDK8/19 activity biomarkers. No collateral binding to any other CDKs was detected with a potency below the cut-off of 1 μM, indicating high selectivity across CDKs in live cells.


Quantifying CDK inhibitor selectivity in live cells.

Wells CI, Vasta JD, Corona CR, Wilkinson J, Zimprich CA, Ingold MR, Pickett JE, Drewry DH, Pugh KM, Schwinn MK, Hwang BB, Zegzouti H, Huber KVM, Cong M, Meisenheimer PL, Willson TM, Robers MB.

Nat Commun. 2020 Jun 2;11(1):2743.

doi: 10.1038/s41467-020-16559-0.

(last updated: 3 May 2022)