ChEMBL ligand: CHEMBL2177390 (GDC-0068, Ipatasertib, Rg-7440, RG-7440, RG7440)
Protein kinase G (PKG) 1/cGMP-dependent protein kinase 1 in Human [ChEMBL: CHEMBL4273] [GtoPdb: 1492] [UniProtKB: Q13976] There should be some charts here, you may need to enable JavaScript!
AKT serine/threonine kinase 1/RAC-alpha serine/threonine-protein kinase in Human [ChEMBL: CHEMBL4282] [GtoPdb: 1479] [UniProtKB: P31749] There should be some charts here, you may need to enable JavaScript!
AKT serine/threonine kinase 2/RAC-beta serine/threonine-protein kinase in Human [ChEMBL: CHEMBL2431] [GtoPdb: 1480] [UniProtKB: P31751] There should be some charts here, you may need to enable JavaScript!
AKT serine/threonine kinase 3/RAC-gamma serine/threonine-protein kinase in Human [ChEMBL: CHEMBL4816] [GtoPdb: 2286] [UniProtKB: Q9Y243] 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
Protein kinase G (PKG) 1/cGMP-dependent protein kinase 1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4273] [GtoPdb: 1492] [UniProtKB: Q13976]
ChEMBL	Inhibition of PRKG1alpha	B	7.01	pIC50	98	nM	IC50	J Med Chem (2012) 55: 8110-8127 [PMID:22934575]
GtoPdb	-	-	7.16	pIC50	69	nM	IC50	J Med Chem (2012) 55: 8110-27 [PMID:22934575]
ChEMBL	Inhibition of PRKG1beta	B	7.16	pIC50	69	nM	IC50	J Med Chem (2012) 55: 8110-8127 [PMID:22934575]
AKT serine/threonine kinase 1/RAC-alpha serine/threonine-protein kinase in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4282] [GtoPdb: 1479] [UniProtKB: P31749]
ChEMBL	Binding affinity to wild-type human partial length AKT1 expressed in bacterial expression system assessed as residual binding level by Kinomescan method	B	9.19	pKd	0.64	nM	Kd	J Med Chem (2021) 64: 18054-18081 [PMID:34855399]
ChEMBL	Inhibition of Akt1 in human LNCAP cells assessed as phosphorylation of PRAS40 at Thr246 after 1.5 hrs	B	6.8	pIC50	157	nM	IC50	J Med Chem (2012) 55: 8110-8127 [PMID:22934575]
ChEMBL	Competitive inhibition of wild-type full-length amino-terminal polyhistidine-tagged human Akt1 expressed in recombinant baculovirus system using fluorescence labeled substrate after 60 mins by fluorescence polarization assay in presence of ATP	B	8.3	pIC50	5	nM	IC50	J Med Chem (2012) 55: 8110-8127 [PMID:22934575]
ChEMBL	Inhibition of Akt1 (unknown origin)	B	8.3	pIC50	5	nM	IC50	Eur J Med Chem (2021) 225: 113749-113749 [PMID:34411892]
ChEMBL	Inhibition of recombinant AKT1 (104 to 480 end residues) (unknown origin) catalytic domain expressed in Sf21 cells using peptide substrate incubated for 1 hr in presence of ATP by caliper off-chip mobility shift assay	B	8.42	pIC50	3.8	nM	IC50	J Med Chem (2022) 65: 8144-8168 [PMID:35679512]
ChEMBL	Covalent-allosteric inhibition of full length N-terminal his6-tagged AKT (2 to 446 residues) (unknown origin) expressed in Spodoptera frugiperda Sf9 cells by HTRF analysis	B	8.46	pIC50	3.5	nM	IC50	Chem Sci (2019) 10: 3573-3585 [PMID:30996949]
ChEMBL	Biochemical Assays: iFLiK and HTRF studies were carried out as described in Z. Fang, J. R. Simard, D. Plenker, H. D. Nguyen, T. Phan, P. Wolle, S. Baumeister, D. Rauh, ACS Chem. Biol. 2015, 10, 279-288.All reagents for HTRF experiments were purchased from Cisbio Bioassays, France. OriginPro 9.1G software (OriginLab Corporation, Northhampton, Mass.) was used for data analysis and data was fit to a sigmoidal dose-response model using the following four-parameter logistic equation:y = A 2 + ( A 2 - A 1 ) ( 1 + ( x IC 50 ) p ) ( 1 ) (A1: bottom asymptote; A2: top asymptote; IC50: half-maximal inhibitory concentration; p: Hill coefficient) Kinetic Characterization of Covalent Probe Compounds:Time-dependent IC50 measurements were performed with activated full-length Akt1 as described under Biochemical assays. Briefly, IC50 values were determined for twelve different incubation times and afterwards plotted versus accordingly. Data was analyzed according to literature procedure as described in B. F. Krippendorff, R. Neuhaus, P. Lienau, A. Reichel, W. Huisinga, J. Biomol. Screen. 2009, 14, 913-923. Ki and kinact were calculated with XLfit (Version 5.4.0.8, IDBS, Munich, Germany) defining the substrate concentration as 250 nM and the corresponding substrate KM as 150 nM.Mass Spectrometry:Purified full-length wtAkt1 was thawed under cold water and diluted to a final concentration of 1 mg/mL in storage buffer (50 mM HEPES, 200 mM NaCl, 10% Glycerol, pH 7.4). 20 μL of the respective mixture were mixed with 2 molar equivalents of the compounds of formulas (1a) and (2a), respectively (10 mM in DMSO); samples containing equal volumes of DMSO were individually prepared for control measurements. Following incubation for thirty minutes on ice, the samples were analyzed by ESI-MS using an Agilent 1100 Series HPLC System connected to a ThermoFinnigan LTQ Linear Ion Trap mass spectrometer. Therefore, 6 μL of sample were injected and separated using a Vydac 214TP C4 5 u column (150 mm×2.1 mm) starting at 20% of solvent B for five minutes followed by a gradient up to 90% of solvent B over 14 min (flow rate 210 μL/min) with 0.1% TFA in water as solvent A and 0.1% TFA in acetonitrile as solvent B. After washing the column for two minutes with 90% of solvent B, the concentration of solvent A was increased to 80% in 1 min and the column was washed for five additional minutes. During the complete experiment, a mass range of 700 to 2000 m/z was scanned and raw data was deconvoluted and analyzed with MagTran and mMass (Version 5.5.0) software.For ESI-MS/MS measurements, samples were denatured, separated via SDS-PAGE followed by staining with Coomassie Brilliant Blue and prepared according to standard tryptic in-gel digest protocols as described in A. Shevchenko, H. Tomas, J. Havlis, J. V. Olsen, M. Mann, Nat. Protoc. 2006, 1, 2856-2860. Subsequently, samples were thawed, dissolved in 20 μL of 0.1% TFA in water, sonicated at room temperature for 15 min, and centrifuged at 15000×g for 1 min shortly before analysis. 3 μL of sample were loaded onto a pre-column cartridge and desalted for 5 min using 0.1% TFA in water as eluent at a flow rate of 30 μL/min. The samples were back-flushed from the pre-column to the nano-HPLC column during the whole analysis. Elution was performed using a gradient starting at 5% B with a final composition of 30% B after 35 min (flow rate 300 nL/min) using 0.1% formic acid in water as eluent A and 0.1% formic acid in acetonitrile as eluent B and a column temperature of 40° C. The nano-HPLC column was washed by increasing the percentage of solvent B to 60% in 5 min and to 95% in additional 5 min, washing the columns for further 5 min, flushing back to starting conditions and equilibration of the system for 14 min. During the complete gradient cycle, a typical TOP10 shot-gun proteomics method for the MS and MS/MS analysis was used.	B	8.59	pIC50	2.6	nM	IC50	US-10550114-B2. Kinase inhibitors and their use in cancer therapy (2020)
ChEMBL	Biochemical Assays: iFLiK and HTRF studies were carried out as described in Z. Fang, J. R. Simard, D. Plenker, H. D. Nguyen, T. Phan, P. Wolle, S. Baumeister, D. Rauh, ACS Chem. Biol. 2015, 10, 279-288.All reagents for HTRF experiments were purchased from Cisbio Bioassays, France. OriginPro 9.1G software (OriginLab Corporation, Northhampton, Mass.) was used for data analysis and data was fit to a sigmoidal dose-response model using the following four-parameter logistic equation:y = A 2 + ( A 2 - A 1 ) ( 1 + ( x IC 50 ) p ) ( 1 ) (A1: bottom asymptote; A2: top asymptote; IC50: half-maximal inhibitory concentration; p: Hill coefficient) Kinetic Characterization of Covalent Probe Compounds:Time-dependent IC50 measurements were performed with activated full-length Akt1 as described under Biochemical assays. Briefly, IC50 values were determined for twelve different incubation times and afterwards plotted versus accordingly. Data was analyzed according to literature procedure as described in B. F. Krippendorff, R. Neuhaus, P. Lienau, A. Reichel, W. Huisinga, J. Biomol. Screen. 2009, 14, 913-923. Ki and kinact were calculated with XLfit (Version 5.4.0.8, IDBS, Munich, Germany) defining the substrate concentration as 250 nM and the corresponding substrate KM as 150 nM.Mass Spectrometry:Purified full-length wtAkt1 was thawed under cold water and diluted to a final concentration of 1 mg/mL in storage buffer (50 mM HEPES, 200 mM NaCl, 10% Glycerol, pH 7.4). 20 μL of the respective mixture were mixed with 2 molar equivalents of the compounds of formulas (1a) and (2a), respectively (10 mM in DMSO); samples containing equal volumes of DMSO were individually prepared for control measurements. Following incubation for thirty minutes on ice, the samples were analyzed by ESI-MS using an Agilent 1100 Series HPLC System connected to a ThermoFinnigan LTQ Linear Ion Trap mass spectrometer. Therefore, 6 μL of sample were injected and separated using a Vydac 214TP C4 5 u column (150 mm×2.1 mm) starting at 20% of solvent B for five minutes followed by a gradient up to 90% of solvent B over 14 min (flow rate 210 μL/min) with 0.1% TFA in water as solvent A and 0.1% TFA in acetonitrile as solvent B. After washing the column for two minutes with 90% of solvent B, the concentration of solvent A was increased to 80% in 1 min and the column was washed for five additional minutes. During the complete experiment, a mass range of 700 to 2000 m/z was scanned and raw data was deconvoluted and analyzed with MagTran and mMass (Version 5.5.0) software.For ESI-MS/MS measurements, samples were denatured, separated via SDS-PAGE followed by staining with Coomassie Brilliant Blue and prepared according to standard tryptic in-gel digest protocols as described in A. Shevchenko, H. Tomas, J. Havlis, J. V. Olsen, M. Mann, Nat. Protoc. 2006, 1, 2856-2860. Subsequently, samples were thawed, dissolved in 20 μL of 0.1% TFA in water, sonicated at room temperature for 15 min, and centrifuged at 15000×g for 1 min shortly before analysis. 3 μL of sample were loaded onto a pre-column cartridge and desalted for 5 min using 0.1% TFA in water as eluent at a flow rate of 30 μL/min. The samples were back-flushed from the pre-column to the nano-HPLC column during the whole analysis. Elution was performed using a gradient starting at 5% B with a final composition of 30% B after 35 min (flow rate 300 nL/min) using 0.1% formic acid in water as eluent A and 0.1% formic acid in acetonitrile as eluent B and a column temperature of 40° C. The nano-HPLC column was washed by increasing the percentage of solvent B to 60% in 5 min and to 95% in additional 5 min, washing the columns for further 5 min, flushing back to starting conditions and equilibration of the system for 14 min. During the complete gradient cycle, a typical TOP10 shot-gun proteomics method for the MS and MS/MS analysis was used.	B	8.7	pIC50	2	nM	IC50	US-10550114-B2. Kinase inhibitors and their use in cancer therapy (2020)
ChEMBL	Inhibition of N-terminal GST-tagged human AKT1 (104 to 408 residues) expressed in baculovirus infected Sf21 cells incubated for 1 hr in presence of ATP by mobility shift assay	B	8.89	pIC50	1.3	nM	IC50	J Med Chem (2021) 64: 12163-12180 [PMID:34375113]
GtoPdb	-	-	9	pIC50	1	nM	IC50	J Med Chem (2012) 55: 8110-27 [PMID:22934575]
AKT serine/threonine kinase 2/RAC-beta serine/threonine-protein kinase in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2431] [GtoPdb: 1480] [UniProtKB: P31751]
ChEMBL	Binding affinity to wild-type human partial length AKT2 expressed in bacterial expression system assessed as residual binding level by Kinomescan method	B	7.46	pKd	35	nM	Kd	J Med Chem (2021) 64: 18054-18081 [PMID:34855399]
ChEMBL	Inhibition of recombinant AKT2 (120 to 481 end residues) (unknown origin) catalytic domain expressed in Sf21 cells using peptide substrate incubated for 1 hr in presence of ATP by caliper off-chip mobility shift assay	B	7.32	pIC50	48	nM	IC50	J Med Chem (2022) 65: 8144-8168 [PMID:35679512]
ChEMBL	Inhibition of full length human Akt2 S474D mutant using GRPRTSSFAEGKK as substrate incubated for 40 mins by scintillation counting method	B	7.49	pIC50	32	nM	IC50	Eur J Med Chem (2020) 189: 112076-112076 [PMID:32007668]
GtoPdb	-	-	7.74	pIC50	18	nM	IC50	J Med Chem (2012) 55: 8110-27 [PMID:22934575]
ChEMBL	Competitive inhibition of wild-type full-length amino-terminal polyhistidine-tagged human Akt2 expressed in recombinant baculovirus system using fluorescence labeled substrate after 60 mins by fluorescence polarization assay in presence of ATP	B	7.74	pIC50	18	nM	IC50	J Med Chem (2012) 55: 8110-8127 [PMID:22934575]
ChEMBL	Inhibition of Akt2 (unknown origin)	B	7.74	pIC50	18	nM	IC50	Eur J Med Chem (2021) 225: 113749-113749 [PMID:34411892]
ChEMBL	Inhibition of N-terminal GST-tagged human AKT2 (120 to 481 residues) expressed in baculovirus infected Sf21 cells incubated for 1 hr in presence of ATP by mobility shift assay	B	8.42	pIC50	3.8	nM	IC50	J Med Chem (2021) 64: 12163-12180 [PMID:34375113]
AKT serine/threonine kinase 3/RAC-gamma serine/threonine-protein kinase in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4816] [GtoPdb: 2286] [UniProtKB: Q9Y243]
ChEMBL	Binding affinity to wild-type human partial length AKT3 expressed in bacterial expression system assessed as residual binding level by Kinomescan method	B	8.6	pKd	2.5	nM	Kd	J Med Chem (2021) 64: 18054-18081 [PMID:34855399]
ChEMBL	Inhibition of full length human Akt3 S472D mutant using GRPRTSSFAEGKK as substrate incubated for 40 mins by scintillation counting method	B	7.59	pIC50	26	nM	IC50	Eur J Med Chem (2020) 189: 112076-112076 [PMID:32007668]
ChEMBL	Inhibition of recombinant N-terminal GST tagged AKT2 (108 to 479 end residues) (unknown origin) catalytic domain expressed in Sf21 cells using peptide substrate incubated for 1 hr in presence of ATP by caliper off-chip mobility shift assay	B	7.66	pIC50	22	nM	IC50	J Med Chem (2022) 65: 8144-8168 [PMID:35679512]
ChEMBL	Competitive inhibition of wild-type full-length amino-terminal polyhistidine-tagged human Akt3 expressed in recombinant baculovirus system using fluorescence labeled Crosstide as substrate after 60 mins by fluorescence polarization assay in presence of ATP	B	8.1	pIC50	8	nM	IC50	J Med Chem (2012) 55: 8110-8127 [PMID:22934575]
ChEMBL	Inhibition of Akt3 (unknown origin)	B	8.1	pIC50	8	nM	IC50	Eur J Med Chem (2021) 225: 113749-113749 [PMID:34411892]
GtoPdb	-	-	8.1	pIC50	8	nM	IC50	J Med Chem (2012) 55: 8110-27 [PMID:22934575]

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]