ChEMBL ligand: CHEMBL3301606 (G-154578, G154578, Glpg-0778, GLPG-0778, GLPG0778, GLPG-0788, Gsk-2586184, GSK-2586184, GSK-2586184A, GSK2586184A, Solcitinib)
tyrosine kinase 2/Non-receptor tyrosine-protein kinase TYK2 in Human [ChEMBL: CHEMBL3553] [GtoPdb: 2269] [UniProtKB: P29597] There should be some charts here, you may need to enable JavaScript!
Janus kinase 1/Tyrosine-protein kinase JAK1 in Human [ChEMBL: CHEMBL2835] [GtoPdb: 2047] [UniProtKB: P23458] There should be some charts here, you may need to enable JavaScript!
Janus kinase 2/Tyrosine-protein kinase JAK2 in Human [ChEMBL: CHEMBL2971] [GtoPdb: 2048] [UniProtKB: O60674] There should be some charts here, you may need to enable JavaScript!
Janus kinase 3/Tyrosine-protein kinase JAK3 in Human [ChEMBL: CHEMBL2148] [GtoPdb: 2049] [UniProtKB: P52333] There should be some charts here, you may need to enable JavaScript!

DB	Assay description	Assay Type	Standard value	Standard parameter	Original value	Original units	Original parameter	Reference
tyrosine kinase 2/Non-receptor tyrosine-protein kinase TYK2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3553] [GtoPdb: 2269] [UniProtKB: P29597]
ChEMBL	Inhibition of TYK2 (unknown origin)	B	6.65	pIC50	225	nM	IC50	Eur J Med Chem (2020) 192: 112155-112155 [PMID:32120325]
ChEMBL	TYK2 Inhibition Assay: Recombinant human TYK2 catalytic domain (amino acids 871-1187; catalog number 08-147) was purchased from Carna biosciences. 5 ng of TYK2 was incubated with 12.5 ug polyGT substrate (Sigma catalog number Pb0275) in kinase reaction buffer (25 mM Hepes pH 7.5, 100 mM NaCl, 0.2 mM Na3VO4, 0.1% NP-40, 0.1 uM non-radioactive ATP, 0.125 uCi 33P-gamma-ATP (GE Healthcare, catalog number AH9968) final concentrations) with or without 5 uL containing test compound or vehicle (DMSO, 1% final concentration), in a total volume of 25 uL, in a polypropylene 96-well plate (Greiner, V-bottom). After 90 min at 30° C., reactions were stopped by adding of 25 uL/well of 150 mM phosphoric acid. All of the terminated kinase reaction was transferred to prewashed (75 mM phosphoric acid) 96 well filter plates (Perkin Elmer catalog number 6005177) using a cell harvester (Perkin Elmer). Plates were washed 6 times with 300 uL per well of a 75 mM phosphoric acid solution and the bottom of the plates was sealed. 40 uL/well of Microscint-20 was added, the top of the plates was sealed and readout was performed using the Topcount (Perkin Elmer). Kinase activity was calculated by subtracting counts per minute (cpm) obtained in the presence of a positive control inhibitor (10 uM staurosporine) from cpm obtained in the presence of vehicle. The ability of a test compound to inhibit this activity was determined as: Percentage inhibition=((cpm determined for sample with test compound present-cpm determined for sample with positive control inhibitor) divided by (cpm determined in the presence of vehicle-cpm determined for sample with positive control inhibitor))*100. Dose dilution series were prepared for the compounds enabling the testing of dose-response effects in the TYK2 assay and the calculation of the IC50 for the compound. Each compound was routinely tested at concentration of 20 uM followed by a 1/3 serial dilution, 8 points (20 uM-6.67 uM-2.22 uM-740 nM-247 nM-82 nM-27 nM-9 nM) in a final concentration of 1% DMSO. When potency of compound series increased, more dilutions were prepared and/or the top concentration was lowered (e.g. 5 uM, 1 uM).	B	6.66	pIC50	219	nM	IC50	US-9505754-B2. Compound useful for the treatment of degenerative and inflammatory diseases (2016)
GtoPdb	-	-	6.66	pIC50	219	nM	IC50	WO2010149771. Novel compound useful for the treatment of degenerative and inflammatory diseases (2010)
Janus kinase 1/Tyrosine-protein kinase JAK1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2835] [GtoPdb: 2047] [UniProtKB: P23458]
ChEMBL	JAK1 Inhibition Assay: Recombinant human JAK1 catalytic domain (amino acids 850-1154; catalog number 08-144) was purchased from Carna Biosciences. 10 ng of JAK1 was incubated with 12.5 μg polyGT substrate (Sigma catalog number P0275) in kinase reaction buffer (15 mM Tris-HCl pH 7.5, 1 mM DTT, 0.01% Tween-20, 10 mM MgCl2, 2 μM non-radioactive ATP, 0.25 μCi 33P-gamma-ATP (GE Healthcare, catalog number AH9968) final concentrations) with or without 5 μL containing test compound or vehicle (DMSO, 1% final concentration), in a total volume of 25 μL, in a polypropylene 96-well plate (Greiner, V-bottom). After 45 min at 30° C., reactions were stopped by adding of 25 μL/well of 150 mM phosphoric acid. All of the terminated kinase reaction was transferred to prewashed (75 mM phosphoric acid) 96 well filter plates (Perkin Elmer catalog number 6005177) using a cell harvester (Perkin Elmer). Plates were washed 6 times with 300 μL per well of a 75 mM phosphoric acid solution and the bottom of the plates was sealed. 40 μL/well of Microscint-20 was added, the top of the plates was sealed and readout was performed using the Topcount (Perkin Elmer). Kinase activity was calculated by subtracting counts per minute (cpm) obtained in the presence of a positive control inhibitor (10 μM staurosporine) from cpm obtained in the presence of vehicle. The ability of a test compound to inhibit this activity was determined as:Percentage inhibition=((cpm determined for sample with test compound present−cpm determined for sample with positive control inhibitor) divided by (cpm determined in the presence of vehicle−cpm determined for sample with positive control inhibitor))*100%.	B	7	pIC50	<100	nM	IC50	US-10206907-B2. Compounds useful for the treatment of degenerative and inflammatory diseases (2019)
ChEMBL	Inhibition of JAK1 (unknown origin)	B	8.01	pIC50	9.8	nM	IC50	Eur J Med Chem (2020) 192: 112155-112155 [PMID:32120325]
GtoPdb	-	-	8.18	pIC50	6.6	nM	IC50	WO2010149771. Novel compound useful for the treatment of degenerative and inflammatory diseases (2010)
ChEMBL	Inhibition Assay: Recombinant human JAK1 (catalytic domain, amino acids 866-1154; catalog number PV4774) was purchased from Invitrogen. 1 ng of JAK1 was incubated with 20 nM Ulight-JAK1(tyr1023) peptide (Perkin Elmer catalog number TRF0121) in kinase reaction buffer (25 mM MOPS pH6.8, 0.016% Brij-35, 8.33 mM MgCl2, 3.33 mM DTT, 7 uM ATP) with or without 4 uL containing test compound or vehicle (DMSO, 1% final concentration), in a total volume of 20 uL, in a white 384 Luminotrac 200 plate (Greiner, catalog number 781075). After 60 min at room temperature, reactions were stopped by adding 20 uL/well of detection mixture (1x detection buffer (Perkin Elmer, catalog number CR97-100C), 0.5 nM Europium-anti-phosphotyrosine (PT66) (Perkin Elmer, catalog number AD0068), 10 mM EDTA). Readout is performed using the Envision with excitation at 320 nm and measuring emission at 615 nm (Perkin Elmer). Kinase activity was calculated by subtracting relative fluorescence units (RFU).	B	8.18	pIC50	6.6	nM	IC50	US-8796457-B2. Compound useful for the treatment of degenerative and inflammatory diseases (2014)
ChEMBL	JAK1 Inhibition Assay: Recombinant human JAK1 (catalytic domain, amino acids 866-1154; catalog number PV4774) was purchased from Invitrogen. 1 ng of JAK1 was incubated with 20 nM Ulight-JAK1(tyr1023) peptide (Perkin Elmer catalog number TRF0121) in kinase reaction buffer (25 mM MOPS pH6.8, 0.016% Brij-35, 8.33 mM MgCl2, 3.33 mM DTT, 7 uM ATP) with or without 4 uL containing test compound or vehicle (DMSO, 1% final concentration), in a total volume of 20 uL, in a white 384 Luminotrac 200 plate (Greiner, catalog number 781075). After 60 min at room temperature, reactions were stopped by adding 20 uL/well of detection mixture (1x detection buffer (Perkin Elmer, catalog number CR97-100C), 0.5 nM Europium-anti-phosphotyrosine (PT66) (Perkin Elmer, catalog number AD0068), 10 mM EDTA). Readout is performed using the Envision with excitation at 320 nm and measuring emission at 615 nm (Perkin Elmer). Kinase activity was calculated by subtracting relative fluorescence units (RFU) obtained in the presence of a positive control inhibitor (10 uM staurosporine) from RFU obtained in the presence of vehicle. The ability of a test compound to inhibit this activity was determined as: Percentage inhibition=((RFU determined for sample with test compound present-RFU determined for sample with positive control inhibitor) divided by (RFU determined in the presence of vehicle-RFU determined for sample with positive control inhibitor))*100. Dose dilution series were prepared for the compounds enabling the testing of dose-response effects in the JAK1 assay and the calculation of the IC50 for the compound. Each compound is routinely tested at concentration of 20 uM followed by a 1/5 serial dilution, 8 points (20 uM-4 uM-800 nM-160 nM-32 nM-6.4 nM-1.28 nM-0.26 nM) in a final concentration of 1% DMSO. When potency of compound series increases, more dilutions are prepared and/or the top concentration are lowered (e.g. 5 uM, 1 uM). The data are expressed as the average IC50 from the assays+standard error of the mean.	B	8.18	pIC50	6.6	nM	IC50	US-9505754-B2. Compound useful for the treatment of degenerative and inflammatory diseases (2016)
Janus kinase 2/Tyrosine-protein kinase JAK2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2971] [GtoPdb: 2048] [UniProtKB: O60674]
ChEMBL	JAK2 Inhibition Assay: Recombinant human JAK2 catalytic domain (amino acids 808-1132; catalog number PV4210) was purchased from Invitrogen. 0.025 mU of JAK2 was incubated with 2.5 μg polyGT substrate (Sigma catalog number P0275) in kinase reaction buffer (5 mM MOPS pH 7.5, 9 mM MgAc, 0.3 mM EDTA, 0.06% Brij and 0.6 mM DTT, 1 μM non-radioactive ATP, 0.25 μCi 33P-gamma-ATP (GE Healthcare, catalog number AH9968) final concentrations) with or without 5 μL containing test compound or vehicle (DMSO, 1% final concentration), in a total volume of 25 μL, in a polypropylene 96-well plate (Greiner, V-bottom). After 90 min at 30° C., reactions were stopped by adding of 25 μL/well of 150 mM phosphoric acid. All of the terminated kinase reaction was transferred to prewashed (75 mM phosphoric acid) 96 well filter plates (Perkin Elmer catalog number 6005177) using a cell harvester (Perkin Elmer). Plates were washed 6 times with 300 μL per well of a 75 mM phosphoric acid solution and the bottom of the plates was sealed. 40 μL/well of Microscint-20 was added, the top of the plates was sealed and readout was performed using the Topcount (Perkin Elmer). Kinase activity was calculated by subtracting counts per minute (cpm) obtained in the presence of a positive control inhibitor (10 μM staurosporine) from cpm obtained in the presence of vehicle. The ability of a test compound to inhibit this activity was determined as:Percentage inhibition=((cpm determined for sample with test compound present−cpm determined for sample with positive control inhibitor) divided by (cpm determined in the presence of vehicle−cpm determined for sample with positive control inhibitor))*100%.	B	6.52	pIC50	300	nM	IC50	US-10206907-B2. Compounds useful for the treatment of degenerative and inflammatory diseases (2019)
ChEMBL	Inhibition of JAK2 (unknown origin)	B	6.97	pIC50	107.8	nM	IC50	Eur J Med Chem (2020) 192: 112155-112155 [PMID:32120325]
GtoPdb	-	-	7.17	pIC50	67	nM	IC50	WO2010149771. Novel compound useful for the treatment of degenerative and inflammatory diseases (2010)
ChEMBL	JAK2 Inhibition Assay: Recombinant human JAK2 (catalytic domain, amino acids 866-1154; catalog number PV4210) was purchased from Invitrogen. 0.0125 mU of JAK2 was incubated with 25 nM Ulight-JAK1(tyr1023) peptide (Perkin Elmer catalog number TRF0121) in kinase reaction buffer (41.66 mM HEPES pH7.0, 0.016% Triton X-100, 12.5 mM MgCl2, 3.33 mM DTT, 7.5M ATP) with or without 4L containing test compound or vehicle (DMSO, 1% final concentration), in a total volume of 20L, in a white 384 Luminotrac 200 plate (Greiner, catalog number 781075). After 60 min at room temperature, reactions were stopped by adding 20L/well of detection mixture (1x detection buffer (Perkin Elmer, catalog number CR97-100C), 0.5 nM Europium-anti-phosphotyrosine (PT66) (Perkin Elmer, catalog number AD0068), 10 mM EDTA). Readout is performed using the Envision with excitation at 320 nm and measuring emission at 615 nm (Perkin Elmer). Kinase activity was calculated by subtracting relative fluorescence units (RFU) obtained in the presence of a positive control inhibitor (10M staurosporine) from RFU obtained in the presence of vehicle. The ability of a test compound to inhibit this activity was determined as: Percentage inhibition=((RFU determined for sample with test compound present-RFU determined for sample with positive control inhibitor) divided by (RFU determined in the presence of vehicle-RFU determined for sample with positive control inhibitor))*100. Dose dilution series are prepared for compound enabling the testing of dose-response effects in the JAK2 assay and the calculation of the IC50 for the compound. Each compound is routinely tested at concentration of 20M followed by a 1/5 serial dilution, 8 points (20M-4M-800 nM-160 nM-32 nM-6.4 nM-1.28 nM-0.26 nM) in a final concentration of 1% DMSO. When potency of compound series increases, more dilutions are prepared and/or the top concentration are lowered (e.g. 5M, 1M). The data are expressed as the average IC50 from the assays standard error of the mean.	B	7.17	pIC50	67	nM	IC50	US-9505754-B2. Compound useful for the treatment of degenerative and inflammatory diseases (2016)
Janus kinase 3/Tyrosine-protein kinase JAK3 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2148] [GtoPdb: 2049] [UniProtKB: P52333]
ChEMBL	Inhibition of JAK3 (unknown origin)	B	6.27	pIC50	539	nM	IC50	Eur J Med Chem (2020) 192: 112155-112155 [PMID:32120325]
ChEMBL	JAK3 Inhibition Assay: Recombinant human JAK3 catalytic domain (amino acids 781-1124; catalog number PV3855) was purchased from Invitrogen. 0.025 mU of JAK3 was incubated with 2.5 ug polyGT substrate (Sigma catalog number Pb0275) in kinase reaction buffer (25 mM Tris pH 7.5, 0.5 mM EGTA, 0.5 mM Na3VO4, 5 mM b-glycerolphosphate, 0.01% Triton X-100, 1 uM non-radioactive ATP, 0.25 uCi 33P-gamma-ATP (GE Healthcare, catalog number AH9968) final concentrations) with or without 5 uL containing test compound or vehicle (DMSO, 1% final concentration), in a total volume of 25 uL, in a polypropylene 96-well plate (Greiner, V-bottom). After 105 min at 30° C., reactions were stopped by adding of 25 uL/well of 150 mM phosphoric acid. All of the terminated kinase reaction was transferred to prewashed (75 mM phosphoric acid) 96 well filter plates (Perkin Elmer catalog number 6005177) using a cell harvester (Perkin Elmer). Plates were washed 6 times with 300 uL per well of a 75 mM phosphoric acid solution and the bottom of the plates was sealed. 40 uL/well of Microscint-20 was added, the top of the plates was sealed and readout was performed using the Topcount (Perkin Elmer). Kinase activity was calculated by subtracting counts per minute (cpm) obtained in the presence of a positive control inhibitor (10 uM staurosporine) from cpm obtained in the presence of vehicle. The ability of a test compound to inhibit this activity was determined as: Percentage inhibition=((cpm determined for sample with test compound present-cpm determined for sample with positive control inhibitor) divided by (cpm determined in the presence of vehicle-cpm determined for sample with positive control inhibitor))*100. Dose dilution series were prepared for the compound enabling the testing of dose-response effects in the JAK3 assay and the calculation of the IC50 for the compound. Each compound was routinely tested at concentration of 20 uM followed by a 1/3 serial dilution, 8 points (20 uM-6.67 uM-2.22 uM-740 nM-247 nM-82 nM-27 nM-9 nM) in a final concentration of 1% DMSO. When potency of the compound series increased, more dilutions were prepared and/or the top concentration was lowered (e.g. 5 uM, 1 uM). The data are expressed as the average IC50 from the assays standard error of the mean.	B	6.39	pIC50	408	nM	IC50	US-9505754-B2. Compound useful for the treatment of degenerative and inflammatory diseases (2016)
GtoPdb	-	-	6.39	pIC50	408	nM	IC50	WO2010149771. Novel compound useful for the treatment of degenerative and inflammatory diseases (2010)
ChEMBL	JAK3 Inhibition Assay: Recombinant human JAK3 catalytic domain (amino acids 781-1124; catalog number PV3855) was purchased from Invitrogen. 0.025 mU of JAK3 was incubated with 2.5 μg polyGT substrate (Sigma catalog number P0275) in kinase reaction buffer (25 mM Tris pH 7.5, 0.5 mM EGTA, 0.5 mM Na3VO4, 5 mM b-glycerophosphate, 0.01% Triton X-100, 1 μM non-radioactive ATP, 0.25 μCi 33P-gamma-ATP (GE Healthcare, catalog number AH9968) final concentrations) with or without 5 μL containing test compound or vehicle (DMSO, 1% final concentration), in a total volume of 25 μL, in a polypropylene 96-well plate (Greiner, V-bottom). After 105 min at 30° C., reactions were stopped by adding of 25 μL/well of 150 mM phosphoric acid. All of the terminated kinase reaction was transferred to prewashed (75 mM phosphoric acid) 96 well filter plates (Perkin Elmer catalog number 6005177) using a cell harvester (Perkin Elmer). Plates were washed 6 times with 300 μL per well of a 75 mM phosphoric acid solution and the bottom of the plates was sealed. 40 μL/well of Microscint-20 was added, the top of the plates was sealed and readout was performed using the Topcount (Perkin Elmer). Kinase activity was calculated by subtracting counts per minute (cpm) obtained in the presence of a positive control inhibitor (10 μM staurosporine) from cpm obtained in the presence of vehicle. The ability of a test compound to inhibit this activity was determined as:Percentage inhibition=((cpm determined for sample with test compound present−cpm determined for sample with positive control inhibitor) divided by (cpm determined in the presence of vehicle−cpm determined for sample with positive control inhibitor))*100%.	B	7	pIC50	<100	nM	IC50	US-10206907-B2. Compounds useful for the treatment of degenerative and inflammatory diseases (2019)

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]