Potency data for probe JNJ-3738 and control JNJ-6240
In vitro potency
| Target name | Target information | Species | Assay description | JNJ-3738 | JNJ-6240 | Publication of assay conditions |
|---|---|---|---|---|---|---|
| CDK7, CCNH, MNAT1 | recombinant human CAK complex (CDK7/cyclin H/MAT1) | Homo sapiens | Biochemical CDK7 assay (KIapp): CDK7 catalyzed, ATP-dependent, phosphorylation of a peptide substrate derived from RNA Pol II (CDK7/9-tide). Coupled via lactate dehydrogenase (LDH) and pyruvate kinase (PK) to lactate and NAD+ production, with concomitant depletion of NADH. Continuous coupled assay measures CDK7 catalytic activity through the diminished absorbance intensity at 340 nm that corresponds with NADH depletion. | KIapp = 31 nM | Slide 28 in 'JNJ-3738 Covalent inhibitor for CDK7.pdf' | |
| CDK7, CCNH, MNAT1 | recombinant human CAK complex (CDK7/cyclin H/MAT1) | Homo sapiens | Biochemical CDK7 assay (kinact/KIapp): CDK7 catalyzed, ATP-dependent, phosphorylation of a peptide substrate derived from RNA Pol II (CDK7/9-tide). Coupled via lactate dehydrogenase (LDH) and pyruvate kinase (PK) to lactate and NAD+ production, with concomitant depletion of NADH. Continuous coupled assay measures CDK7 catalytic activity through the diminished absorbance intensity at 340 nm that corresponds with NADH depletion. | kinact/KIapp = 2603 M-1s-1 | Slide 28 in 'JNJ-3738 Covalent inhibitor for CDK7.pdf' | |
| CDK7 | Homo sapiens | Fluorescent end-point assay: KM ATP app ~30 µM | IC50 ~ 200 nM |
In cell potency
| Target name | Target information | Species | Assay description | JNJ-3738 | JNJ-6240 | Publication of assay conditions |
|---|---|---|---|---|---|---|
| CDK7 | A549 parental (wt) | Homo sapiens | In vitro pharmacodynamics: p-RNA pol II Ser-5 (wtCDK7): QuantiGene Plex Gene Expression Assay: downregulation at the messenger level of superenhancer driven genes. The level of phosphorylation of Serine 5 on RNA pol II Vs totalRNA pol II is determined. The level of p-RNA pol II Ser-5 detected and stained with primary (a specific rabbit Phospho-Rpb1 CTD (Ser5) antibody) and secondary antibodies (a goat anti rabbit Alexa Fluor 488 conjugated secondary antibody) after 3 hrs incubation with probe. | IC50 = 58 nM | IC50 > 10 µM | Slide 29 in 'JNJ-3738 Covalent inhibitor for CDK7.pdf' |
| CDK7 (C312S) | A549 cells overexpressing mutant CDK7 (C312S) | Homo sapiens | In vitro pharmacodynamics: p-RNA pol II Ser-5 (CDK7 C312S): QuantiGene Plex Gene Expression Assay: downregulation at the messenger level of superenhancer driven genes. The level of phosphorylation of Serine 5 on RNA pol II Vs totalRNA pol II is determined. The level of p-RNA pol II Ser-5 detected and stained with primary (a specific rabbit Phospho-Rpb1 CTD (Ser5) antibody) and secondary antibodies (a goat anti rabbit Alexa Fluor 488 conjugated secondary antibody) after 3 hrs incubation with probe. | IC50 > 10 µM | IC50 > 10 µM | Slide 29 in 'JNJ-3738 Covalent inhibitor for CDK7.pdf' |
| CDK7 | OCI-AML3 cells containing CDK7 WT | Homo sapiens | Antiproliferation assay using OCI-AML3 cells (wtCDK7): Proliferation inhibition measured by CTG after 4 days compound treatment | IC50 = 4 nM | Slide 29 in 'JNJ-3738 Covalent inhibitor for CDK7.pdf' | |
| CDK7 (C312S) | OCI-AML3 cells containing mutant CDK7 (C312S) | Homo sapiens | Antiproliferation assay using OCI-AML3 cells (CDK7 C312S): Proliferation inhibition measured by CTG after 4 days compound treatment | IC50 = 3.98 µM | Slide 29 in 'JNJ-3738 Covalent inhibitor for CDK7.pdf' |