crozbaciclib [Ligand Id: 12076] activity data from GtoPdb and ChEMBL
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| ChEMBL ligand: CHEMBL4277900 (Crozbaciclib) |
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| DB | Assay description | Assay Type | Standard value | Standard parameter | Original value | Original units | Original parameter | Reference |
|---|---|---|---|---|---|---|---|---|
| cyclin dependent kinase 1/CDK1/Cyclin A in Human (target type: PROTEIN COMPLEX) [ChEMBL: CHEMBL3038467] [GtoPdb: 1961] [UniProtKB: P06493, P20248] | ||||||||
| ChEMBL | Inhibition of CDK1/Cyclin-A2 (unknown origin) using histone-H1 as substrate after 90 mins by ADP-Glo assay | B | 5.57 | pIC50 | 2707 | nM | IC50 | Eur J Med Chem (2018) 144: 1-28 [PMID:29247857] |
| cyclin dependent kinase 2/CDK2/Cyclin-E2 in Human (target type: PROTEIN COMPLEX) [ChEMBL: CHEMBL4523633] [GtoPdb: 1973] [UniProtKB: O96020, P24941] | ||||||||
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 5.57 | pIC50 | 2707 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 5.57 | pIC50 | 2707 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 5.57 | pIC50 | 2707 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 5.57 | pIC50 | 2707 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| cyclin dependent kinase 6/CDK6/cyclin D3 in Human (target type: PROTEIN COMPLEX) [ChEMBL: CHEMBL2111448] [GtoPdb: 1978] [UniProtKB: P30281, Q00534] | ||||||||
| GtoPdb | Inhibition of CDK6/cyclin D3 in vitro | - | 9 | pIC50 | 1 | nM | IC50 | Eur J Med Chem (2018) 144: 1-28 [PMID:29247857] |
| ChEMBL | Inhibition of CDK6/Cyclin-D3 (unknown origin) using histoneH1 as substrate after 90 mins by ADP-Glo assay | B | 9 | pIC50 | 1 | nM | IC50 | Eur J Med Chem (2018) 144: 1-28 [PMID:29247857] |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 9.14 | pIC50 | 0.73 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 9.14 | pIC50 | 0.73 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 9.14 | pIC50 | 0.73 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 9.14 | pIC50 | 0.73 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| cyclin dependent kinase 1/Cyclin-dependent kinase 1/ G1/S-specific cyclin-D3 in Human (target type: PROTEIN COMPLEX) [ChEMBL: CHEMBL4296065] [GtoPdb: 1961] [UniProtKB: P06493, P30281] | ||||||||
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 8.27 | pIC50 | 5.36 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 8.27 | pIC50 | 5.36 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 8.27 | pIC50 | 5.36 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| cyclin dependent kinase 2/Cyclin-dependent kinase 2/cyclin E1 in Human (target type: PROTEIN COMPLEX) [ChEMBL: CHEMBL1907605] [GtoPdb: 1973] [UniProtKB: P24864, P24941] | ||||||||
| ChEMBL | Inhibition of CDK2/Cyclin-E1 (unknown origin) using histone-H1 as substrate after 90 mins by ADP-Glo assay | B | 5.63 | pIC50 | 2321 | nM | IC50 | Eur J Med Chem (2018) 144: 1-28 [PMID:29247857] |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 5.63 | pIC50 | 2321 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| cyclin dependent kinase 4/Cyclin-dependent kinase 4/cyclin D1 in Human (target type: PROTEIN COMPLEX) [ChEMBL: CHEMBL1907601] [GtoPdb: 1976] [UniProtKB: P11802, P24385] | ||||||||
| GtoPdb | Inhibition of CDK4/cyclin D1 in vitro | - | 8.52 | pIC50 | 3 | nM | IC50 | Eur J Med Chem (2018) 144: 1-28 [PMID:29247857] |
| ChEMBL | Inhibition of CDK4/Cyclin-D1 (unknown origin) using histone-H1 as substrate after 90 mins by ADP-Glo assay | B | 8.52 | pIC50 | 3 | nM | IC50 | Eur J Med Chem (2018) 144: 1-28 [PMID:29247857] |
| cyclin dependent kinase 4/Cyclin-dependent kinase 4/G1/S-specific cyclin-E1 in Human (target type: PROTEIN COMPLEX) [ChEMBL: CHEMBL3885554] [GtoPdb: 1976] [UniProtKB: P11802, P24864] | ||||||||
| GtoPdb | Inhibition of CDK4/cyclin D1 in vitro | - | 8.52 | pIC50 | 3 | nM | IC50 | Eur J Med Chem (2018) 144: 1-28 [PMID:29247857] |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 8.58 | pIC50 | 2.62 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| mitogen-activated protein kinase kinase kinase 14/Mitogen-activated protein kinase kinase kinase 14 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5888] [GtoPdb: 2074] [UniProtKB: Q99558] | ||||||||
| ChEMBL | Inhibition of recombinant human GST-tagged NIK expressed in baculovirus expression system using ser/thr-7 as substrate preincubated for 15 mins followed by ATP addition measured after 60 mins | B | 7.35 | pIC50 | 45 | nM | IC50 | Eur J Med Chem (2018) 144: 1-28 [PMID:29247857] |
| Pim-1 proto-oncogene, serine/threonine kinase/Serine/threonine-protein kinase pim-1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2147] [GtoPdb: 2158] [UniProtKB: P11309] | ||||||||
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 7.42 | pIC50 | 38.3 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| ChEMBL | Kinase Inhibitory Activity Assay: 1.2.2 Preparation of kinase solution: Kinase solutions having the concentrations required for each reaction system were prepared from 100 ng/μl kinase stock solution and the 1× kinase reaction buffer. 1.2.3 Preparation of the test compound solution and control LY2835219 solution: (1) Preparation of control LY2835219 solution a. 1 μl of 10 mM standard substance stock solution was taken and added to 9 μl of kinase reaction buffer I and mixed well; then 90 μl of kinase reaction buffer I was added and mixed well; then 100 μl of kinase reaction buffer I was added and mixed well. The final concentration was 50 μM. b. 40 μl of kinase reaction buffer II was added into B2 to B10 of a 96-well plate, and 50 μl of the above solution was added into B1; c. 10 μl of solution was taken from well B1, added into B2, and mixed well; then 10 μl of the resulting solution was taken and added into B3, and dilution was carried out in sequence to B9, so as to obtain control solutions that are diluted by 5-fold sequentially. (2) Preparation of test compound solution: a. Test compound solutions at a certain concentration were taken, respectively, diluted with kinase reaction buffer I to a final concentration of 50 μM compound solution; b. 40 μl of kinase reaction buffer II was added into H2 to H10 of the 96-well plate; and 50 μl of the above solution was added into H1; c. 10 μl of solution was taken from well H1, added into H2, and mixed well; then 10 μl of the resulting solution was taken and added into H3, and dilution was carried out in sequence to H9, so as to obtain test compound solutions that are diluted by 5-fold sequentially. 1.2.4 Preparation of a mixed solution of reaction substrate and ATP: a. Preparation of ATP solution: 200 μl of 0.1 mM ATP solution: 2 μl of 10 mM ATP was added to 198 μl of kinase reaction buffer I; 300 μl of 50 μM ATP solution: 150 μl of kinase reaction buffer I was added to 150 μl of the above 0.1 mM ATP solution; b. Preparation of 300 μl of reaction substrate solution: 150 μl 1 μg/μl reaction substrate stock solution was added to 150 μl kinase reaction buffer I, and mixed well; c. The above a/b solutions were mixed to obtain mixed solutions, respectively. | B | 7.42 | pIC50 | 38.3 | nM | IC50 | US-11091476-B2. Protein kinase inhibitors, preparation method and medical use thereof (2021) |
| Janus kinase 3/Tyrosine-protein kinase JAK3 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2148] [GtoPdb: 2049] [UniProtKB: P52333] | ||||||||
| ChEMBL | Inhibition of recombinant human GST-tagged JAK3 cytoplasmic domain (781 to 1124 residues) expressed in baculovirus expression system using ULight-JAK-1 peptide as substrate preincubated for 15 mins followed by ATP addition measured after 90 mins | B | 6.81 | pIC50 | 154 | nM | IC50 | Eur J Med Chem (2018) 144: 1-28 [PMID:29247857] |
ChEMBL data shown on this page come from version 36:
Zdrazil B, Felix E, Hunter F, Manners EJ, Blackshaw J, Corbett S, de Veij M, Ioannidis H, Lopez DM, Mosquera JF, Magarinos MP, Bosc N, Arcila R, Kizilören T, Gaulton A, Bento AP, Adasme MF, Monecke P, Landrum GA, Leach AR. (2024). The ChEMBL Database in 2023: a drug discovery platform spanning multiple bioactivity data types and time periods. Nucleic Acids Res., 52(D1). DOI: 10.1093/nar/gkad1004. [EPMCID:10767899] [PMID:37933841]
Davies M, Nowotka M, Papadatos G, Dedman N, Gaulton A, Atkinson F, Bellis L, Overington JP. (2015) 'ChEMBL web services: streamlining access to drug discovery data and utilities.' Nucleic Acids Res., 43(W1). DOI: 10.1093/nar/gkv352. [EPMCID:25883136]
