lovingscience/chembl-v37-raw-activities huggingface dataset:
Compound (SMILES): CN(C)Cc1ccc(Nc2ncc3c(=O)c(C(N)=O)c(N)n(CC(F)(F)F)c3n2)cc1
InChIKey: BHBAMVLIFUDMOO-UHFFFAOYSA-N
Target: Calcium/calmodulin-dependent protein kinase type II subunit delta (SINGLE PROTEIN), Homo sapiens
UniProt: Q13557
Assay: In Vitro Inhibition Assay: The kinase activity of CaMKII beta is evaluated by means of a radioactive test on the recombinant CaMKII beta enzyme. The amount of ATP-gamma 33P incorporated into the specific substrate Autocamtide-2 during its phosphorylation by CaMKII beta is measured. The effect of the products is quantified by the concentration of product that inhibits the total activity of CaMKII beta by 50% (50% inhibitory concentration=IC50). For the determination of the IC50 values, the product is diluted in 100% of DMSO to obtain a 10 mM stock solution. The concentration range tested during the radioactive test ranges from 3 to 10 000 nM with a final concentration in the test of 1% DMSO. This concentration range may, for the most powerful compounds, be extended to 0.1 nM. On the day of the operation, 5 ul of the compounds are deposited in each well of a 96-well plate at 10 times the concentration of that to be tested. Each concentration is tested in duplicate on the same plate.
Measurement: IC50 = 663 nM (pChEMBL 6.179999828338623)
Descriptors: MW 435.41, ALogP 1.84, HBA 8, HBD 3, PSA 132.16, QED 0.54
Compound (SMILES): CCn1c(N)c(C(N)=O)c(=O)c2cnc(Nc3ccc(N4CCC(N5CCCC5)CC4)c(Cl)c3)nc21
InChIKey: ZRQLWBZFRLHXBV-UHFFFAOYSA-N
Target: Calcium/calmodulin-dependent protein kinase type II subunit delta (SINGLE PROTEIN), Homo sapiens
UniProt: Q13557
Assay: In Vitro Inhibition Assay: The kinase activity of CaMKII beta is evaluated by means of a radioactive test on the recombinant CaMKII beta enzyme. The amount of ATP-gamma 33P incorporated into the specific substrate Autocamtide-2 during its phosphorylation by CaMKII beta is measured. The effect of the products is quantified by the concentration of product that inhibits the total activity of CaMKII beta by 50% (50% inhibitory concentration=IC50). For the determination of the IC50 values, the product is diluted in 100% of DMSO to obtain a 10 mM stock solution. The concentration range tested during the radioactive test ranges from 3 to 10 000 nM with a final concentration in the test of 1% DMSO. This concentration range may, for the most powerful compounds, be extended to 0.1 nM. On the day of the operation, 5 ul of the compounds are deposited in each well of a 96-well plate at 10 times the concentration of that to be tested. Each concentration is tested in duplicate on the same plate.
Measurement: IC50 = 8 nM (pChEMBL 8.100000381469727)
Descriptors: MW 511.03, ALogP 2.95, HBA 9, HBD 3, PSA 135.4, QED 0.46
Compound (SMILES): CCn1c(N)c(C(N)=O)c(=O)c2cnc(Nc3ccc4c(c3)CCNC4)nc21
InChIKey: ZQUKGJSMCUOBNN-UHFFFAOYSA-N
Target: Calcium/calmodulin-dependent protein kinase type II subunit delta (SINGLE PROTEIN), Homo sapiens
UniProt: Q13557
Assay: In Vitro Inhibition Assay: The kinase activity of CaMKII beta is evaluated by means of a radioactive test on the recombinant CaMKII beta enzyme. The amount of ATP-gamma 33P incorporated into the specific substrate Autocamtide-2 during its phosphorylation by CaMKII beta is measured. The effect of the products is quantified by the concentration of product that inhibits the total activity of CaMKII beta by 50% (50% inhibitory concentration=IC50). For the determination of the IC50 values, the product is diluted in 100% of DMSO to obtain a 10 mM stock solution. The concentration range tested during the radioactive test ranges from 3 to 10 000 nM with a final concentration in the test of 1% DMSO. This concentration range may, for the most powerful compounds, be extended to 0.1 nM. On the day of the operation, 5 ul of the compounds are deposited in each well of a 96-well plate at 10 times the concentration of that to be tested. Each concentration is tested in duplicate on the same plate.
Measurement: IC50 = 37 nM (pChEMBL 7.429999828338623)
Descriptors: MW 379.42, ALogP 0.88, HBA 8, HBD 4, PSA 140.95, QED 0.53
Compound (SMILES): CCn1c(N)c(C(N)=O)c(=O)c2cnc(Nc3ccc(OC4CCN(C)CC4)cc3)nc21
InChIKey: TVQRLARWAXWMMR-UHFFFAOYSA-N
Target: Calcium/calmodulin-dependent protein kinase type II subunit delta (SINGLE PROTEIN), Homo sapiens
UniProt: Q13557
Assay: In Vitro Inhibition Assay: The kinase activity of CaMKII beta is evaluated by means of a radioactive test on the recombinant CaMKII beta enzyme. The amount of ATP-gamma 33P incorporated into the specific substrate Autocamtide-2 during its phosphorylation by CaMKII beta is measured. The effect of the products is quantified by the concentration of product that inhibits the total activity of CaMKII beta by 50% (50% inhibitory concentration=IC50). For the determination of the IC50 values, the product is diluted in 100% of DMSO to obtain a 10 mM stock solution. The concentration range tested during the radioactive test ranges from 3 to 10 000 nM with a final concentration in the test of 1% DMSO. This concentration range may, for the most powerful compounds, be extended to 0.1 nM. On the day of the operation, 5 ul of the compounds are deposited in each well of a 96-well plate at 10 times the concentration of that to be tested. Each concentration is tested in duplicate on the same plate.
Measurement: IC50 = 118 nM (pChEMBL 6.929999828338623)
Descriptors: MW 437.5, ALogP 1.71, HBA 9, HBD 3, PSA 141.39, QED 0.53
Compound (SMILES): CCn1c(N)c(C(N)=O)c(=O)c2cnc(Nc3cccc(CN(C)C)c3)nc21
InChIKey: DXHKCCUMIZFGOJ-UHFFFAOYSA-N
Target: Calcium/calmodulin-dependent protein kinase type II subunit delta (SINGLE PROTEIN), Homo sapiens
UniProt: Q13557
Assay: In Vitro Inhibition Assay: The kinase activity of CaMKII beta is evaluated by means of a radioactive test on the recombinant CaMKII beta enzyme. The amount of ATP-gamma 33P incorporated into the specific substrate Autocamtide-2 during its phosphorylation by CaMKII beta is measured. The effect of the products is quantified by the concentration of product that inhibits the total activity of CaMKII beta by 50% (50% inhibitory concentration=IC50). For the determination of the IC50 values, the product is diluted in 100% of DMSO to obtain a 10 mM stock solution. The concentration range tested during the radioactive test ranges from 3 to 10 000 nM with a final concentration in the test of 1% DMSO. This concentration range may, for the most powerful compounds, be extended to 0.1 nM. On the day of the operation, 5 ul of the compounds are deposited in each well of a 96-well plate at 10 times the concentration of that to be tested. Each concentration is tested in duplicate on the same plate.
Measurement: IC50 = 268 nM (pChEMBL 6.570000171661377)
Descriptors: MW 381.44, ALogP 1.3, HBA 8, HBD 3, PSA 132.16, QED 0.59
Compound (SMILES): CCc1cc(N2CCN(C3CC3)CC2)ccc1Nc1ncc2c(=O)c(C(N)=O)c(N)n(CC)c2n1
InChIKey: CQXBKUODUUZWDT-UHFFFAOYSA-N
Target: Calcium/calmodulin-dependent protein kinase type II subunit delta (SINGLE PROTEIN), Homo sapiens
UniProt: Q13557
Assay: In Vitro Inhibition Assay: The kinase activity of CaMKII beta is evaluated by means of a radioactive test on the recombinant CaMKII beta enzyme. The amount of ATP-gamma 33P incorporated into the specific substrate Autocamtide-2 during its phosphorylation by CaMKII beta is measured. The effect of the products is quantified by the concentration of product that inhibits the total activity of CaMKII beta by 50% (50% inhibitory concentration=IC50). For the determination of the IC50 values, the product is diluted in 100% of DMSO to obtain a 10 mM stock solution. The concentration range tested during the radioactive test ranges from 3 to 10 000 nM with a final concentration in the test of 1% DMSO. This concentration range may, for the most powerful compounds, be extended to 0.1 nM. On the day of the operation, 5 ul of the compounds are deposited in each well of a 96-well plate at 10 times the concentration of that to be tested. Each concentration is tested in duplicate on the same plate.
Measurement: IC50 = 396 nM (pChEMBL 6.400000095367432)
Descriptors: MW 476.59, ALogP 2.08, HBA 9, HBD 3, PSA 135.4, QED 0.47
Compound (SMILES): COc1cc(C2CCNCC2)ccc1Nc1ncc2c(=O)c(C(N)=O)c(N)n(-c3ccccc3)c2n1
InChIKey: WMZGFAFYMYUVFX-UHFFFAOYSA-N
Target: Calcium/calmodulin-dependent protein kinase type II subunit delta (SINGLE PROTEIN), Homo sapiens
UniProt: Q13557
Assay: In Vitro Inhibition Assay: The kinase activity of CaMKII beta is evaluated by means of a radioactive test on the recombinant CaMKII beta enzyme. The amount of ATP-gamma 33P incorporated into the specific substrate Autocamtide-2 during its phosphorylation by CaMKII beta is measured. The effect of the products is quantified by the concentration of product that inhibits the total activity of CaMKII beta by 50% (50% inhibitory concentration=IC50). For the determination of the IC50 values, the product is diluted in 100% of DMSO to obtain a 10 mM stock solution. The concentration range tested during the radioactive test ranges from 3 to 10 000 nM with a final concentration in the test of 1% DMSO. This concentration range may, for the most powerful compounds, be extended to 0.1 nM. On the day of the operation, 5 ul of the compounds are deposited in each well of a 96-well plate at 10 times the concentration of that to be tested. Each concentration is tested in duplicate on the same plate.
Measurement: IC50 = 39 nM (pChEMBL 7.409999847412109)
Descriptors: MW 485.55, ALogP 2.68, HBA 9, HBD 4, PSA 150.18, QED 0.33
Compound (SMILES): COc1cc(C2CCNCC2)ccc1Nc1ncc2c(=O)c(C(N)=O)c(N)n(CCO)c2n1
InChIKey: KACVMLZSIGLBSY-UHFFFAOYSA-N
Target: Calcium/calmodulin-dependent protein kinase type II subunit delta (SINGLE PROTEIN), Homo sapiens
UniProt: Q13557
Assay: In Vitro Inhibition Assay: The kinase activity of CaMKII beta is evaluated by means of a radioactive test on the recombinant CaMKII beta enzyme. The amount of ATP-gamma 33P incorporated into the specific substrate Autocamtide-2 during its phosphorylation by CaMKII beta is measured. The effect of the products is quantified by the concentration of product that inhibits the total activity of CaMKII beta by 50% (50% inhibitory concentration=IC50). For the determination of the IC50 values, the product is diluted in 100% of DMSO to obtain a 10 mM stock solution. The concentration range tested during the radioactive test ranges from 3 to 10 000 nM with a final concentration in the test of 1% DMSO. This concentration range may, for the most powerful compounds, be extended to 0.1 nM. On the day of the operation, 5 ul of the compounds are deposited in each well of a 96-well plate at 10 times the concentration of that to be tested. Each concentration is tested in duplicate on the same plate.
Measurement: IC50 = 57 nM (pChEMBL 7.239999771118164)
Descriptors: MW 453.5, ALogP 0.68, HBA 10, HBD 5, PSA 170.41, QED 0.35
Compound (SMILES): COCCCOc1cc(CN(C(=O)[C@H]2CNCCO2)C2CC2)cc2ccccc12
InChIKey: PXKWFPHRFQPVMA-JOCHJYFZSA-N
Target: Renin (SINGLE PROTEIN), Homo sapiens
UniProt: P00797
Assay: Inhibition Assay: A renin substrate of synthetic peptide (Nma-KHPFH LVIHK(Dnp)-NH2) and test compound were mixed, and fluorescence intensity was assayed using a fluorophotometer before starting an enzymatic reaction (exciting wavelength: 340 nm, measuring wavelength: 460 nm). Recombinant human renin was added and the mixture was incubated at 37 C. for an hour, and the fluorescence intensity was measured after the reaction using using a fluorophotometer (exciting wavelength: 340 nm, measuring wavelength: 460 nm). Renin activity was evaluated on the ground of fluorescence intensity which was obtained by deduction of the intensity before the reaction from the intensity after the reaction, and 50% inhibitory concentration (IC50) was calculated from renin activities under the existence of various concentration of the tested compound.
Measurement: IC50 = 30.2 nM (pChEMBL 7.519999980926514)
Descriptors: MW 398.5, ALogP 2.73, HBA 5, HBD 1, PSA 60.03, QED 0.66
Compound (SMILES): COCCCn1cc(CN(C(=O)[C@H]2CNCCO2)C2CC2)c2ccccc21
InChIKey: OHELVELDNLQVJI-HXUWFJFHSA-N
Target: Renin (SINGLE PROTEIN), Homo sapiens
UniProt: P00797
Assay: Inhibition Assay: A renin substrate of synthetic peptide (Nma-KHPFH LVIHK(Dnp)-NH2) and test compound were mixed, and fluorescence intensity was assayed using a fluorophotometer before starting an enzymatic reaction (exciting wavelength: 340 nm, measuring wavelength: 460 nm). Recombinant human renin was added and the mixture was incubated at 37 C. for an hour, and the fluorescence intensity was measured after the reaction using using a fluorophotometer (exciting wavelength: 340 nm, measuring wavelength: 460 nm). Renin activity was evaluated on the ground of fluorescence intensity which was obtained by deduction of the intensity before the reaction from the intensity after the reaction, and 50% inhibitory concentration (IC50) was calculated from renin activities under the existence of various concentration of the tested compound.
Measurement: IC50 = 13 nM (pChEMBL 7.889999866485596)
Descriptors: MW 371.48, ALogP 2.16, HBA 5, HBD 1, PSA 55.73, QED 0.72
Compound (SMILES): Cc1c(-c2cc(Nc3cc4n(n3)CCN(C)C4)c(=O)[nH]n2)cccc1N1Cc2cc(C(C)(C)C)sc2C1=O
InChIKey: ADIGCBHSQFHVRD-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 4000 nM (pChEMBL 5.400000095367432)
Descriptors: MW 529.67, ALogP 4.65, HBA 8, HBD 2, PSA 99.15, QED 0.4
Compound (SMILES): Cc1c(-c2cc(Nc3cc4n(n3)CCN(C)C4)c(=O)[nH]n2)cccc1N1Cc2ccc(C(C)(C)C)cc2C1=O
InChIKey: LNTTWCMWTOIRLF-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 14 nM (pChEMBL 7.849999904632568)
Descriptors: MW 523.64, ALogP 4.59, HBA 7, HBD 2, PSA 99.15, QED 0.41
Compound (SMILES): Cc1c(-c2cc(Nc3cc(C)n(C)n3)c(=O)[nH]n2)cccc1N1Cc2cc(C(C)(C)C)sc2C1=O
InChIKey: GUOFQNFYXCMSED-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 3 nM (pChEMBL 8.520000457763672)
Descriptors: MW 488.62, ALogP 5.05, HBA 7, HBD 2, PSA 95.91, QED 0.42
Compound (SMILES): Cc1c(-c2cc(Nc3cc(C4CC4)[nH]n3)c(=O)[nH]n2)cccc1N1Cc2cc(C(C)(C)C)sc2C1=O
InChIKey: YGKZJAKJFAGODW-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 4 nM (pChEMBL 8.399999618530273)
Descriptors: MW 500.63, ALogP 5.61, HBA 6, HBD 3, PSA 106.77, QED 0.33
Compound (SMILES): Cc1cc(Nc2cc(-c3ccc(F)c(N4Cc5cc(C(C)(C)C)sc5C4=O)c3)n[nH]c2=O)nn1C
InChIKey: XDOHTJZKHHRFGR-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 9 nM (pChEMBL 8.050000190734863)
Descriptors: MW 492.58, ALogP 4.88, HBA 7, HBD 2, PSA 95.91, QED 0.42
Compound (SMILES): Cc1cc(Nc2cc(-c3cccc(N4Cc5cc(C(C)(C)C)sc5C4=O)c3C)n[nH]c2=O)no1
InChIKey: LDIWSMQZSQUVTD-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 18 nM (pChEMBL 7.75)
Descriptors: MW 475.57, ALogP 5.3, HBA 7, HBD 2, PSA 104.12, QED 0.41
Compound (SMILES): CC(C)(C)c1cc2c(s1)C(=O)N(c1nc(-c3cc(Nc4cc(C5CC5)[nH]n4)c(=O)[nH]n3)ccc1F)C2
InChIKey: FIZJDZYXSLJGCM-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 35 nM (pChEMBL 7.460000038146973)
Descriptors: MW 505.58, ALogP 4.83, HBA 7, HBD 3, PSA 119.66, QED 0.36
Compound (SMILES): Cc1cc(Nc2cc(-c3ccc(F)c(N4Cc5cc(C(C)(C)C)sc5C4=O)n3)n[nH]c2=O)nn1C
InChIKey: DYZRFQHAROFHKL-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 45 nM (pChEMBL 7.349999904632568)
Descriptors: MW 493.57, ALogP 4.28, HBA 8, HBD 2, PSA 108.8, QED 0.44
Compound (SMILES): Cc1c(-c2cc(Nc3nc4c(s3)CNCC4)c(=O)[nH]n2)cccc1N1Cc2cc(C(C)(C)C)sc2C1=O
InChIKey: SVJLSEOIBZWLLQ-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 65 nM (pChEMBL 7.190000057220459)
Descriptors: MW 532.7, ALogP 5.11, HBA 8, HBD 3, PSA 103.01, QED 0.34
Compound (SMILES): Cc1c(-c2cc(Nc3cc(CN4CC(F)(F)C4)n(C)n3)c(=O)[nH]n2)cccc1N1Cc2cc(C(C)(C)C)sc2C1=O
InChIKey: FXWCPRZBMYBHJX-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 5 nM (pChEMBL 8.300000190734863)
Descriptors: MW 579.68, ALogP 5.19, HBA 8, HBD 2, PSA 99.15, QED 0.33
Compound (SMILES): CN1CCn2nc(Nc3cc(-c4cccc(N5Cc6cc(C(C)(C)C)sc6C5=O)c4CO)nn(C)c3=O)cc2C1
InChIKey: VZDDUZAFBGBYFG-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 55 nM (pChEMBL 7.260000228881836)
Descriptors: MW 559.7, ALogP 3.84, HBA 10, HBD 2, PSA 108.52, QED 0.38
Compound (SMILES): CN1CCn2nc(Nc3cc(-c4cccc(N5Cc6cc(C(C)(C)C)sc6C5=O)c4CO)n[nH]c3=O)cc2C1
InChIKey: PBIGUDXXZKZSMW-UHFFFAOYSA-N
Target: Tyrosine-protein kinase BTK (SINGLE PROTEIN), Homo sapiens
UniProt: Q06187
Assay: Biochemical Assay: A generalized procedure for a standard biochemical Btk Kinase Assay that can be used to test Formula I compounds is as follows. A master mix minus Btk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta- glycerophosphate, 2 mM dithiothreitol, 0.1 mM Na3V04, 10 mM MgCl2), 0.5 ul Promega PTK Biotinylated peptide substrate 2, and 0.01% BSA. A master mix plus Btk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ul PTK Biotinylated peptide substrate 2, 0.01% BSA, and 100 ng/well (0.06 mU/well) Btk enzyme. Btk enzyme is prepared as follows: full length human wildtype Btk (accession number NM-000061) with a C-terminal V5 and 6x His tag was subcloned into pFastBac vector for making baculo virus carrying this epitope-tagged Btk. Generation of baculovirus is done based on Invitrogen's instructions detailed in its published protocol "Bac-to-Bac Baculovirus Expression Systems".
Measurement: IC50 = 48 nM (pChEMBL 7.320000171661377)
Descriptors: MW 545.67, ALogP 3.83, HBA 9, HBD 3, PSA 119.38, QED 0.35
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2nc3cccnc3cc2-c2cnn(CCO)c2)nc1
InChIKey: SIODGJOUFXQXGM-QAPCUYQASA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 0.23 nM (pChEMBL 9.640000343322754)
Descriptors: MW 417.47, ALogP 3.07, HBA 8, HBD 1, PSA 95.18, QED 0.47
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2nc(C)ncc2-c2cnn(C)c2)nc1
InChIKey: HHKWMTRMYWMUDH-CJNGLKHVSA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 0.25 nM (pChEMBL 9.600000381469727)
Descriptors: MW 351.41, ALogP 2.77, HBA 7, HBD 0, PSA 74.95, QED 0.68
Compound (SMILES): Cn1cc(-c2cc3ccncc3nc2OC[C@H]2C[C@@H]2c2ccc3ncccc3n2)cn1
InChIKey: GUHJHNGFKCUGHF-AEFFLSMTSA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 0.04 nM (pChEMBL 10.399999618530273)
Descriptors: MW 408.47, ALogP 4.16, HBA 7, HBD 0, PSA 78.61, QED 0.43
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2nc(C)ncc2/C=C/c2cccnc2)nc1
InChIKey: OFFGURFOHFNSOB-OANMOWAGSA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 0.6 nM (pChEMBL 9.220000267028809)
Descriptors: MW 374.44, ALogP 3.94, HBA 6, HBD 0, PSA 70.02, QED 0.62
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2nc3cccnc3cc2C2CCCN(C(C)=O)C2)nc1
InChIKey: KDJBOXFYCVDLKX-FLXSOZOKSA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 2.5 nM (pChEMBL 8.600000381469727)
Descriptors: MW 432.52, ALogP 3.94, HBA 6, HBD 0, PSA 77.44, QED 0.59
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2nc(C)ncc2CN2CCC(F)(F)C2)nc1
InChIKey: KBDHBQFXPCRQDT-PBHICJAKSA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 60.8 nM (pChEMBL 7.21999979019165)
Descriptors: MW 390.43, ALogP 3.21, HBA 6, HBD 0, PSA 60.37, QED 0.72
Compound (SMILES): Cc1ncc(-c2cnn(C)c2)c(OC[C@H]2C[C@@H]2c2ccn(C)n2)n1
InChIKey: SBMWOGGNDJAYMO-OCCSQVGLSA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 3.2 nM (pChEMBL 8.489999771118164)
Descriptors: MW 324.39, ALogP 2.1, HBA 7, HBD 0, PSA 70.65, QED 0.72
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2nc(C)ncc2-c2ccc(C)nc2)nc1
InChIKey: RHMDAALVGLZJPI-AEFFLSMTSA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 0.8 nM (pChEMBL 9.100000381469727)
Descriptors: MW 362.43, ALogP 3.74, HBA 6, HBD 0, PSA 70.02, QED 0.67
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2nc(C)ncc2-c2nnc(C3CC3)s2)nc1
InChIKey: AHQGHUJWEMWMPD-HIFRSBDPSA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 4.2 nM (pChEMBL 8.380000114440918)
Descriptors: MW 395.49, ALogP 3.77, HBA 8, HBD 0, PSA 82.91, QED 0.6
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2nc(C)c(C)nc2-c2cncc(OC)c2)nc1
InChIKey: XWTSGLRGJLFIKB-APWZRJJASA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 0.22 nM (pChEMBL 9.65999984741211)
Descriptors: MW 392.46, ALogP 3.75, HBA 7, HBD 0, PSA 79.25, QED 0.61
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2ncc(C)cc2-c2cnn3c2CCC3)nc1
InChIKey: GHQCBMSEFZPBLC-WBVHZDCISA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 0.34 nM (pChEMBL 9.470000267028809)
Descriptors: MW 376.46, ALogP 3.79, HBA 6, HBD 0, PSA 62.06, QED 0.66
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2nc(C)ncc2C2=CCC(O)CC2)nc1
InChIKey: XZMMWKAYVRDZIF-AGPBCMBSSA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 0.15 nM (pChEMBL 9.819999694824219)
Descriptors: MW 367.45, ALogP 3.3, HBA 6, HBD 1, PSA 77.36, QED 0.84
Compound (SMILES): COc1ccc([C@H]2C[C@@H]2COc2nc(C)ncc2-c2csc(C)n2)nc1
InChIKey: NXLDNCSPMSJZLL-HIFRSBDPSA-N
Target: cAMP and cAMP-inhibited cGMP 3',5'-cyclic phosphodiesterase 10A (SINGLE PROTEIN), Homo sapiens
UniProt: Q9Y233
Assay: Fluorescence Polarization Assay: In a typical experiment the PDE10 inhibitory activity of the compounds of the present invention was determined in accordance with the following experimental method. PDE10A2 was amplified from human fetal brain cDNA (Clontech, Mountain View, Calif.) using a forward primer corresponding to nucleotides 56-77 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716), containing a Kozak consensus sequence, and a reverse primer corresponding to nucleotides 2406-2413 of human PDE10A2 (Accession No. AF127480, Genbank Identifier 4894716). Amplification with Easy-A polymerase (Stratagene, La Jolla, Calif.) was 95 C. for 2 minutes followed by thirty three cycles of 95 C. for 40 seconds, 55 C. for 30 seconds, and 72 C. for 2 minutes 48 seconds. Final extension was 72 C. for 7 minutes. The PCR product was TA cloned into pcDNA3.2-TOPO (Invitrogen, Carlsbad, Calif.) according to standard protocol. AD293 cells with 70-80% confluency were transiently transfected with human PDE10A.
Measurement: Ki = 0.78 nM (pChEMBL 9.109999656677246)
Descriptors: MW 368.46, ALogP 3.8, HBA 7, HBD 0, PSA 70.02, QED 0.66
Compound (SMILES): NC1(c2ccc(-c3nc4ncccn4c3-c3ccccc3)cc2)CCC1
InChIKey: LMBQMEOEJDKMKM-UHFFFAOYSA-N
Target: RAC-alpha serine/threonine-protein kinase (SINGLE PROTEIN), Homo sapiens
UniProt: P31749
Assay: TR-FRET Assay: For the assay 50 nl of a 100 fold concentrated solution of the test compound in DMSO was pipetted into a black low volume 384 well microtiter plate (Greiner Bio-One, Frickenhausen, Germany), 2 ul of a solution of Akt1 in assay buffer [50 mM TRIS/HCl pH 7.5, 5 mM MgCl2, 1 mM dithiothreitol, 0.02% (v/v) Triton X-100 (Sigma)] were added and the mixture was incubated for 15 min at 22 C. to allow prebinding of the test compounds to the enzyme before the start of the kinase reaction. Then the kinase reaction was started by the addition of 3 ul of a solution of adenosine-tri-phosphate (ATP, 16.7 uM=>final conc. in the 5 ul assay volume is 10 uM) and substrate (1.67 uM=>final conc. in the 5 ul assay volume is 1 uM) in assay buffer and the resulting mixture was incubated for a reaction time of 60 min at 22 C. The concentration of Akt1 in the assay was adjusted depending of the activity of the enzyme lot.
Measurement: IC50 = 348 nM (pChEMBL 6.460000038146973)
Descriptors: MW 340.43, ALogP 4.4, HBA 4, HBD 1, PSA 56.21, QED 0.6