Assay Detail
Binding
CHEMBL5734784
Review assay metadata, readout intent, target linkage, and publication context from the same page.
The DYRKIA kinase assay: The DYRKIA kinase assay was run using the Ser/Thr 18 peptide Z-lyte assay kit according to manufacturer's instructions (Life Technologiesa Division of Thermo-Fisher). This is a non-radioactive assay using fluorescence resonance energy transfer (FRET) between coumarin and fluorescein to detect kinase activity which is represented as a ratio of coumarin emission/fluorescein emission.Briefly, recombinant DYRKIA kinase, ATP and Ser/Thr peptide 18 were prepared in 1× Kinase buffer to final concentrations of 0.19 μg/mL, 30 μM, and 4 μM respectively. The mixture was allowed to incubate with the representative compounds for one hour at room temperature. All reactions were performed in duplicate. Unphosphorylated (0% Control) and phosphorylated (100% control) forms of Ser/Thr 18 served as control reactions. Additionally, an 11-point dose-response curve of Staurosporine (1 uM top) was run to serve as a positive compound control.After incubation, Development Reagent A was diluted in Development Buffer then added to the reaction and allowed to further incubate for one hour at room temperature. The plate was read at Ex 400 Em 455 to detect the coumarin signal and Ex 400 Em 520 to measure the signal (EnVision Multilabel Plate Reader, PerkinElmer).The Emission ratio (Em) was calculated as a ratio of the coumarin (C) emission signal (at 445 nm)/Fluorescein (F) emission signal (at 520 nm). The percent phosphorylation was then calculated using the following formula: [1−((Em ratio×F100%)−C100%)/((C0%−C100%)+(Em ratio×(F100%−F0%)))]. Dose-response curves were generated and inhibitory concentration (IC50) values were calculated using non-linear regression curve fit in the Dotmatics' Studies Software (Bishops Stortford, UK).
732
Total Activities
235
Compounds Tested
3
Activity Types
0
Assay Parameters
Assay Information
| Assay Type | Binding |
| Organism | Homo sapiens |
| Confidence | 9 — Direct single protein target |
| Curated By | Autocuration |
Target
Dual specificity tyrosine-phosphorylation-regulated kinase 1A (CHEMBL2292) ↗| Type | SINGLE PROTEIN |
| Organism | Homo sapiens |
Publication
Isoquinolin-3-yl carboxamides and preparation and use thereof
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| EC50 | 244 | 8.44 | 9.40 |
| kon | 244 | - | - |
| k_off | 244 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL5895786 | — | — | 6 | 9.40 |
| CHEMBL5915695 | — | — | 3 | 9.22 |
| CHEMBL4748909 | — | — | 3 | 9.15 |
| CHEMBL5928969 | — | — | 3 | 9.15 |
| CHEMBL5966908 | — | — | 6 | 9.10 |
| CHEMBL5998719 | — | — | 3 | 9.10 |
| CHEMBL4786449 | — | — | 3 | 9.10 |
| CHEMBL5913310 | — | — | 3 | 9.10 |
| CHEMBL5865581 | — | — | 3 | 9.05 |
| CHEMBL5997594 | — | — | 3 | 9.05 |
| CHEMBL6055907 | — | — | 3 | 9.05 |
| CHEMBL5742439 | — | — | 3 | 9.05 |
| CHEMBL4800211 | — | — | 3 | 9.05 |
| CHEMBL4748484 | — | — | 3 | 9.05 |
| CHEMBL6024816 | — | — | 3 | 9.00 |
| CHEMBL5975914 | — | — | 3 | 9.00 |
| CHEMBL4744439 | — | — | 3 | 9.00 |
| CHEMBL5839411 | — | — | 3 | 9.00 |
| CHEMBL4784318 | CIRTUVIVINT | 1.0 | 15 | 9.00 |
| CHEMBL5889809 | — | — | 3 | 8.96 |
| CHEMBL5917165 | — | — | 3 | 8.96 |
| CHEMBL4783628 | — | — | 3 | 8.96 |
| CHEMBL4741472 | — | — | 3 | 8.96 |
| CHEMBL5857068 | — | — | 3 | 8.96 |
| CHEMBL4758853 | — | — | 3 | 8.96 |
| CHEMBL5980669 | — | — | 3 | 8.96 |
| CHEMBL5748878 | — | — | 3 | 8.96 |
| CHEMBL6018949 | — | — | 3 | 8.96 |
| CHEMBL5999029 | — | — | 3 | 8.96 |
| CHEMBL5810561 | — | — | 3 | 8.92 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL5895786 | — | EC50 | = | 0.4 | nM | 9.40 |
| CHEMBL5915695 | — | EC50 | = | 0.6 | nM | 9.22 |
| CHEMBL5928969 | — | EC50 | = | 0.7 | nM | 9.15 |
| CHEMBL4748909 | — | EC50 | = | 0.7 | nM | 9.15 |
| CHEMBL5998719 | — | EC50 | = | 0.8 | nM | 9.10 |
| CHEMBL5913310 | — | EC50 | = | 0.8 | nM | 9.10 |
| CHEMBL4786449 | — | EC50 | = | 0.8 | nM | 9.10 |
| CHEMBL5966908 | — | EC50 | = | 0.8 | nM | 9.10 |
| CHEMBL4800211 | — | EC50 | = | 0.9 | nM | 9.05 |
| CHEMBL6055907 | — | EC50 | = | 0.9 | nM | 9.05 |
| CHEMBL4748484 | — | EC50 | = | 0.9 | nM | 9.05 |
| CHEMBL5997594 | — | EC50 | = | 0.9 | nM | 9.05 |
| CHEMBL5865581 | — | EC50 | = | 0.9 | nM | 9.05 |
| CHEMBL5742439 | — | EC50 | = | 0.9 | nM | 9.05 |
| CHEMBL6024816 | — | EC50 | = | 1.0 | nM | 9.00 |
| CHEMBL5975914 | — | EC50 | = | 1.0 | nM | 9.00 |
| CHEMBL4744439 | — | EC50 | = | 1.0 | nM | 9.00 |
| CHEMBL4784318 | CIRTUVIVINT | EC50 | = | 1.0 | nM | 9.00 |
| CHEMBL5839411 | — | EC50 | = | 1.0 | nM | 9.00 |
| CHEMBL5889809 | — | EC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5980669 | — | EC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5917165 | — | EC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5748878 | — | EC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5999029 | — | EC50 | = | 1.1 | nM | 8.96 |
| CHEMBL4758853 | — | EC50 | = | 1.1 | nM | 8.96 |
| CHEMBL4783628 | — | EC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5857068 | — | EC50 | = | 1.1 | nM | 8.96 |
| CHEMBL4741472 | — | EC50 | = | 1.1 | nM | 8.96 |
| CHEMBL6018949 | — | EC50 | = | 1.1 | nM | 8.96 |
| CHEMBL5884745 | — | EC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5785021 | — | EC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5810561 | — | EC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5855019 | — | EC50 | = | 1.2 | nM | 8.92 |
| CHEMBL4784318 | CIRTUVIVINT | EC50 | = | 1.2 | nM | 8.92 |
| CHEMBL4760089 | — | EC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5937147 | — | EC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5805255 | — | EC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5982202 | — | EC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5966908 | — | EC50 | = | 1.2 | nM | 8.92 |
| CHEMBL5961034 | — | EC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5759195 | — | EC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5969499 | — | EC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5947188 | — | EC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5759930 | — | EC50 | = | 1.3 | nM | 8.89 |
| CHEMBL4780597 | — | EC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5979160 | — | EC50 | = | 1.3 | nM | 8.89 |
| CHEMBL5962060 | — | EC50 | = | 1.3 | nM | 8.89 |
| CHEMBL4747563 | — | EC50 | = | 1.4 | nM | 8.85 |
| CHEMBL5770327 | — | EC50 | = | 1.4 | nM | 8.85 |
| CHEMBL4784318 | CIRTUVIVINT | EC50 | = | 1.4 | nM | 8.85 |