venetoclax [Ligand Id: 8318] activity data from GtoPdb and ChEMBL
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| ChEMBL ligand: CHEMBL3137309 (Abt199, ABT-199, GDC-0199, RG-7601, RG7601, Venclexta, Venclyxto, Venetoclax) |
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| DB | Assay description | Assay Type | Standard value | Standard parameter | Original value | Original units | Original parameter | Reference |
|---|---|---|---|---|---|---|---|---|
| BCL2 apoptosis regulator/Apoptosis regulator Bcl-2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4860] [GtoPdb: 2844] [UniProtKB: P10415] | ||||||||
| ChEMBL | Binding affinity to human full-length N-terminal His6-tagged prephosphorylated Bcl2 (2 to 206 residues) expressed in Escherichia coli S12 extract by isothermal titration calorimetry | B | 6.15 | pKd | 705 | nM | Kd | J Med Chem (2020) 63: 13733-13744 [PMID:33197310] |
| ChEMBL | Binding affinity to His-tagged Bcl-2 G101V mutant (unknown origin) by SPR method | B | 7.54 | pKd | 29 | nM | Kd | J Med Chem (2024) 67: 7836-7858 [PMID:38695063] |
| ChEMBL | Binding Assay: Table 1: Binding to Bcl-2 proteins Bcl-2, and Bcl-XL was assessed using the Bcl2scan™ platform: T7 phage strains displaying BCL2 proteins were grown in parallel in 24-well blocks in an E. coli host derived from the BL21 strain. E. coli were grown to log-phase and infected with T7 phage from a frozen stock (multiplicity of infection=0.4) and incubated with shaking at 32° C. until lysis (90-150 minutes). The lysates were centrifuged (5,000×g) and filtered (0.2 μm) to remove cell debris. Streptavidin-coated magnetic beads were treated with biotinylated BIM peptide ligand for 30 minutes at room temperature to generate affinity resins for BCL2 assays. The liganded beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific phage binding. Binding reactions were assembled by combining BCL2 proteins, liganded affinity beads, and test compounds in 1× binding buffer (20% SeaBlock, 0.17×PBS, 0.05% Tween 20, 6 mM DTT). Test compounds were prepared as 100× stocks in 100% DMSO. Kds were determined using an 11-point 3-fold compound dilution series with one DMSO control point. All compounds for Kd measurements were distributed by acoustic transfer in 100% DMSO. The compounds were then diluted directly into the assays such that the final concentration of DMSO was 0.9%. All reactions performed in polypropylene 384-well plates. Each was a final volume of 0.02 ml. The assay plates were incubated at room temperature with shaking for 1 hour and the affinity beads were washed with wash buffer (lx PBS, 0.05% Tween 20). The beads were then re-suspended in elution buffer (lx PBS, 0.05% Tween 20, 2 μM non-biotinylated affinity ligand) and incubated at room temperature with shaking for 30 minutes. | B | 8 | pKd | <10 | nM | Kd | US-11318134-B2. Benzamide compounds (2022) |
| ChEMBL | Binding Assay: Table 1: Binding to Bcl-2 proteins Bcl-2, and Bcl-XL was assessed using the Bcl2scan™ platform: T7 phage strains displaying BCL2 proteins were grown in parallel in 24-well blocks in an E. coli host derived from the BL21 strain. E. coli were grown to log-phase and infected with T7 phage from a frozen stock (multiplicity of infection=0.4) and incubated with shaking at 32° C. until lysis (90-150 minutes). The lysates were centrifuged (5,000×g) and filtered (0.2 μm) to remove cell debris. Streptavidin-coated magnetic beads were treated with biotinylated BIM peptide ligand for 30 minutes at room temperature to generate affinity resins for BCL2 assays. The liganded beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific phage binding. Binding reactions were assembled by combining BCL2 proteins, liganded affinity beads, and test compounds in 1× binding buffer (20% SeaBlock, 0.17×PBS, 0.05% Tween 20, 6 mM DTT). Test compounds were prepared as 100× stocks in 100% DMSO. Kds were determined using an 11-point 3-fold compound dilution series with one DMSO control point. All compounds for Kd measurements were distributed by acoustic transfer in 100% DMSO. The compounds were then diluted directly into the assays such that the final concentration of DMSO was 0.9%. All reactions performed in polypropylene 384-well plates. Each was a final volume of 0.02 ml. The assay plates were incubated at room temperature with shaking for 1 hour and the affinity beads were washed with wash buffer (lx PBS, 0.05% Tween 20). | B | 8 | pKd | <10 | nM | Kd | US-11344546-B2. Benzamide compounds (2022) |
| ChEMBL | Bcl-2 Protein Family Binding Assay: Binding to Bcl-2 proteins Bcl-2, and Bcl-XL was assessed using the Bcl2scan™ platform: T7 phage strains displaying BCL2 proteins were grown in parallel in 24-well blocks in an E. coli host derived from the BL21 strain. E. coliwere grown to log-phase and infected with T7 phage from a frozen stock (multiplicity of infection=0.4) and incubated with shaking at 32° C. until lysis (90-150 minutes). The lysates were centrifuged (5,000×g) and filtered (0.2 μm) to remove cell debris. Streptavidin-coated magnetic beads were treated with biotinylated BIM peptide ligand for 30 minutes at room temperature to generate affinity resins for BCL2 assays. The liganded beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific phage binding. Binding reactions were assembled by combining BCL2 proteins, liganded affinity beads, and test compounds in 1× binding buffer (20% SeaBlock, 0.17×PBS, 0.05% Tween 20, 6 mM DTT). Test compounds were prepared as 100× stocks in 100% DMSO. Kds were determined using an 11-point 3-fold compound dilution series with one DMSO control point. All compounds for Kd measurements were distributed by acoustic transfer in 100% DMSO. The compounds were then diluted directly into the assays such that the final concentration of DMSO was 0.9%. All reactions performed in polypropylene 384-well plates. Each was a final volume of 0.02 ml. The assay plates were incubated at room temperature with shaking for 1 hour and the affinity beads were washed with wash buffer (lx PBS, 0.05% Tween 20). The beads were then re-suspended in elution buffer (lx PBS, 0.05% Tween 20, 2 μM non-biotinylated affinity ligand) and incubated at room temperature with shaking for 30 minutes. | B | 8 | pKd | <10 | nM | Kd | US-11590126-B2. Benzamide compounds (2023) |
| ChEMBL | Binding affinity to human full-length N-terminal His6-tagged Bcl2 (2 to 206 residues) expressed in Escherichia coli S12 extract by isothermal titration calorimetry | B | 8.51 | pKd | 3.1 | nM | Kd | J Med Chem (2020) 63: 13733-13744 [PMID:33197310] |
| ChEMBL | Binding affinity to recombinant Bcl-2 G101V mutant (unknown origin) using peptide probe incubated for 2 hrs by fluorescence based assay | B | 7.01 | pKi | 98 | nM | Ki | Eur J Med Chem (2022) 232: 114184-114184 [PMID:35182816] |
| ChEMBL | Binding affinity to recombinant Bcl-2 D103V mutant (unknown origin) using peptide probe incubated for 2 hrs by fluorescence based assay | B | 7.23 | pKi | 59 | nM | Ki | Eur J Med Chem (2022) 232: 114184-114184 [PMID:35182816] |
| ChEMBL | Binding affinity to recombinant Bcl-2 D103E mutant (unknown origin) using peptide probe incubated for 2 hrs by fluorescence based assay | B | 7.77 | pKi | 17 | nM | Ki | Eur J Med Chem (2022) 232: 114184-114184 [PMID:35182816] |
| ChEMBL | Binding affinity to recombinant wild-type Bcl-2 (unknown origin) using peptide probe incubated for 2 hrs by fluorescence based assay | B | 8.92 | pKi | 1.2 | nM | Ki | Eur J Med Chem (2022) 232: 114184-114184 [PMID:35182816] |
| ChEMBL | Binding affinity to full length human Bcl-2 expressed in Escherichia coli BL21 (DE3) assessed as inhibition constant preincubated for 30 mins followed by 5-FAM-QEDIIIINIARHLAQVGDSMD-RSIPPG tracer addition and measured after 20 mins by fluorescence polarization assay | B | 9 | pKi | <1 | nM | Ki | J Med Chem (2021) 64: 10260-10285 [PMID:34228434] |
| ChEMBL | Inhibition of FAM-Bid binding to human BCL2 expressed in Escherichia coli BL21 after 30 mins by fluorescence polarization assay | B | 9 | pKi | <1 | nM | Ki | ACS Med Chem Lett (2016) 7: 1185-1190 [PMID:27994761] |
| ChEMBL | Binding affinity to BCL2 (unknown origin) incubated for 30 mins by TR-FRET assay | B | 10 | pKi | <0.1 | nM | Ki | Medchemcomm (2016) 7: 778-787 |
| ChEMBL | Time Resolved-Fluorescence Resonance Energy Transfer (TR-FRET) Assay: The inhibition constant (Ki) is the dissociation constant of an enzyme-inhibitor complex or a protein/small molecule complex, wherein the small molecule is inhibiting binding of one protein to another protein or peptide. Where the Ki for a compound is represented as > (greater than) a certain numerical value, it is intended to mean that the binding affinity value (e.g., for Bcl-XL) is greater than the limits of detection of the assay used. Where the binding selectivity ratio for a compound is represented as > (greater than) a certain numerical value, it is intended to mean that the selectivity of a particular compound for Bcl-2 over Bcl-XL is at least as great as the number indicated. Where the Ki for a compound is represented as < (less than) a certain numerical value, it is intended to mean that the binding affinity value (e.g., for Bcl-2) is lower than the limit of detection of the assay used. Inhibition constants were determined using Wang's equation (Wang Z-X). | B | 10.44 | pKi | 0.04 | nM | Ki | US-9125913-B2. Bcl-2-selective apoptosis-inducing agents for the treatment of cancer and immune diseases (2015) |
| ChEMBL | Time Resolved-Fluorescence Resonance Energy Transfer (TR-FRET) Assay: The inhibition constant (Ki) is the dissociation constant of an enzyme-inhibitor complex or a protein/small molecule complex, wherein the small molecule is inhibiting binding of one protein to another protein or peptide. Where the Ki for a compound is represented as > (greater than) a certain numerical value, it is intended to mean that the binding affinity value (e.g., for Bcl-XL) is greater than the limits of detection of the assay used. Where the binding selectivity ratio for a compound is represented as > (greater than) a certain numerical value, it is intended to mean that the selectivity of a particular compound for Bcl-2 over Bcl-XL is at least as great as the number indicated. Where the Ki for a compound is represented as < (less than) a certain numerical value, it is intended to mean that the binding affinity value (e.g., for Bcl-2) is lower than the limit of detection of the assay used. Inhibition constants were determined using Wang's equation (Wang Z-X). | B | 10.44 | pKi | 0.04 | nM | Ki | US-9125913-B2. Bcl-2-selective apoptosis-inducing agents for the treatment of cancer and immune diseases (2015) |
| ChEMBL | Inhibition of wild-type BCL-2 (unknown origin) expressed in Escherichia coli BL21 cells using biotinylated BIMBH3 or BAXBH3 peptide by surface plasmon resonance assay | B | 10.74 | pKi | 0.02 | nM | Ki | J Med Chem (2020) 63: 928-943 [PMID:31580668] |
| GtoPdb | Note that this Ki is below the detection limit of the assay. | - | 11 | pKi | <0.01 | nM | Ki |
US8580794. Apoptosis-inducing agents for the treatment of cancer and immune and autoimmune diseases. (2013); Nat Med (2013) 19: 202-8 [PMID:23291630] |
| ChEMBL | Binding affinity to BCL-W (unknown origin) assessed as inhibition constant by fluorescence polarization binding affinity assay | B | 11 | pKi | <0.01 | nM | Ki | J Med Chem (2024) 67: 10795-10830 [PMID:38913996] |
| ChEMBL | Binding affinity to Bcl-2 (unknown origin) assessed as dissociation constant incubated for 1 hrs by TR-FRET assay | B | 11 | pKi | <0.01 | nM | Ki | Sci Transl Med (2015) 7: null-null [PMID:25787766] |
| ChEMBL | Time Resolved-Fluorescence Resonance Energy Transfer (TR-FRET) Assay: Representative compounds were serially diluted in dimethyl sulfoxide (DMSO) starting at 50 uM (2x starting concentration; 10% DMSO) and 10 uL were transferred into a 384-well plate. Then 10 uL of a protein/probe/antibody mix was added to each well at final concentrations listed in TABLE 1. Protein:GST-Bcl-2, Probe: F-Bak Peptide Probe Acetyl-GQVGRQLAIIGDK(6-FAM)INR-amide(SEQ ID NO: 1), Protein(nM): 1, Probe (nM): 100, Antibody: Tb-anit-GST, Antibody (nm): 1. The samples are then mixed on a shaker for 1 minute and incubated for an additional 3 hours at room temperature. For each assay, the probe/antibody and protein/probe/antibody were included on each assay plate as negative and positive controls, respectively. Fluorescence was measured on the ENVISION plate reader (Perkin Elmer) using a 340/35 nm excitation filter and 520/525 (F-Bak peptide) and 495/510 nm (Tb-labeled anti-Histidine antibody) emission filters. | B | 11 | pKi | <0.01 | nM | Ki | US-9174982-B2. Apoptosis-inducing agents for the treatment of cancer and immune and autoimmune diseases (2015) |
| ChEMBL | TR-FRET (Time-Resolved Fluorescence-Resonance-Energy-Transfer) assay: TR-FRET (Time-Resolved Fluorescence-Resonance-Energy-Transfer) assay. | B | 11 | pKi | <0.01 | nM | Ki | US-10213433-B2. Solid dispersions containing an apoptosis-inducing agent (2019) |
| ChEMBL | TBDTR-FRET (Time-Resolved Fluorescence-Resonance-Energy-Transfer) Assay: The inhibition constant (Ki) for binding of representative compounds to Bcl-2 protein, as determined by a TR-FRET (Time-Resolved Fluorescence-Resonance-Energy-Transfer) assay. | B | 11 | pKi | <0.01 | nM | Ki | US-11369599-B2. Melt-extruded solid dispersions containing an apoptosis-inducing agent (2022) |
| ChEMBL | Binding affinity to Bcl-2 (unknown origin) by FRET assay | B | 11 | pKi | <0.01 | nM | Ki | Bioorg Med Chem Lett (2016) 26: 2105-2114 [PMID:26988306] |
| ChEMBL | TBDTR-FRET (Time-Resolved Fluorescence-Resonance-Energy-Transfer) Assay: The inhibition constant (Ki) for binding of representative compounds to Bcl-2 protein, as determined by a TR-FRET (Time-Resolved Fluorescence-Resonance-Energy-Transfer) assay. | B | 11 | pKi | <0.01 | nM | Ki | US-11369599-B2. Melt-extruded solid dispersions containing an apoptosis-inducing agent (2022) |
| ChEMBL | Time Resolved-Fluorescence Resonance Energy Transfer (TR-FRET) Assay: Representative compounds were serially diluted in dimethyl sulfoxide (DMSO) starting at 50 uM (2x starting concentration; 10% DMSO) and 10 uL were transferred into a 384-well plate. Then 10 uL of a protein/probe/antibody mix was added to each well at final concentrations listed in TABLE 1. Protein:GST-Bcl-2, Probe: F-Bak Peptide Probe Acetyl-GQVGRQLAIIGDK(6-FAM)INR-amide(SEQ ID NO: 1), Protein(nM): 1, Probe (nM): 100, Antibody: Tb-anit-GST, Antibody (nm): 1. The samples are then mixed on a shaker for 1 minute and incubated for an additional 3 hours at room temperature. For each assay, the probe/antibody and protein/probe/antibody were included on each assay plate as negative and positive controls, respectively. Fluorescence was measured on the ENVISION plate reader (Perkin Elmer) using a 340/35 nm excitation filter and 520/525 (F-Bak peptide) and 495/510 nm (Tb-labeled anti-Histidine antibody) emission filters. | B | 11 | pKi | <0.01 | nM | Ki | US-9174982-B2. Apoptosis-inducing agents for the treatment of cancer and immune and autoimmune diseases (2015) |
| ChEMBL | Binding affinity to Bcl-2 (unknown origin) by TR-FRET assay | B | 11 | pKi | <0.01 | nM | Ki | J Med Chem (2017) 60: 821-838 [PMID:27749061] |
| ChEMBL | Inhibition of Bcl-2 (unknown origin) by TR-FRET assay | B | 11 | pKi | <0.01 | nM | Ki | Eur J Med Chem (2018) 146: 471-482 [PMID:29407973] |
| ChEMBL | Inhibition of Bcl2 (unknown origin) | B | 11 | pKi | <0.01 | nM | Ki | J Med Chem (2018) 61: 6421-6467 [PMID:29620890] |
| ChEMBL | Inhibition of Bcl2 (unknown origin) | B | 11 | pKi | <0.01 | nM | Ki | J Med Chem (2018) 61: 2636-2651 [PMID:28926247] |
| ChEMBL | Inhibition of Bcl2 (unknown origin) | B | 11 | pKi | <0.01 | nM | Ki | Eur J Med Chem (2019) 177: 63-75 [PMID:31129454] |
| ChEMBL | Inhibition of BCL2 (unknown origin) by TR-FRET assay | B | 11 | pKi | <0.01 | nM | Ki | J Med Chem (2020) 63: 11420-11435 [PMID:32539387] |
| ChEMBL | Displacement of fluorescent-labeled BID-BH3 peptide from human BCL-2 expressed in Escherichia coli BL21 (DE3) assessed as inhibition constant by fluorescence polarization-based competitive binding assay | B | 11 | pKi | <0.01 | nM | Ki | Eur J Med Chem (2022) 236: 114327-114327 [PMID:35385805] |
| ChEMBL | Binding affinity to Bcl-2 (unknown origin) assessed as inhibition constant by TR-FRET assay | B | 11 | pKi | <0.01 | nM | Ki | Eur J Med Chem (2020) 201: 112446-112446 [PMID:32563811] |
| ChEMBL | Binding affinity to Bcl-2 (unknown origin) pre-incubated for 30 mins followed by addition FAM-peptide further incubated for 20 mins by fluorescence polarization-based binding assay | B | 4 | pIC50 | >100000 | nM | IC50 | J Med Chem (2024) 67: 13925-13958 [PMID:39121336] |
| ChEMBL | Inhibition of FAM-Bid peptide binding to Bcl2 (unknown origin) by fluorescence polarization assay | B | 6.64 | pIC50 | 230 | nM | IC50 | Bioorg Med Chem Lett (2019) 29: 349-352 [PMID:30594434] |
| ChEMBL | Inhibition of FAM-Bim peptide binding to human Bcl-2 (2 to 206) measured after 30 mins by fluorescence polarization assay | B | 7 | pIC50 | >100 | nM | IC50 | Eur J Med Chem (2021) 220: 113452-113452 [PMID:33906046] |
| ChEMBL | Bcl-2-G101V Biochemical Assay : Selected compounds disclosed herein were tested for blocking of Bcl-2-G101 protein with its ligand in an assay based on time-resolved fluorescence resonance energy transfer methodology. 0.05 nM of Recombinant human Bcl-2-G101V protein was pre-incubated with a serial dilution of compounds disclosed herein (maximum concentration is 10 ÎĽM, 4-fold serially diluted, 10 points; or maximum concentration is 1 uM, 3-fold serially diluted, 10 points) at room temperature for 0.5 hour in an assay buffer containing 20 mM potassium phosphate buffer, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.05% Tween-20, 0.01% BSA. Then 5 nM of the FITC labeled Bak peptide Ac-GQVGRQLAIIGDK(FITC)INR-amide and Mab Anti-6His Tb cryptate Gold was added to plate and further incubated at room temperature for 1 hour. The TR-FRET signals (ex337 nm, em490 nm/520 nm) were read on BMG PHERAstar FSX instrument. The inhibition percentage of Bcl-2-G101V interaction with its ligand in presence of increasing concentrations of compounds was calculated based on the ratio of fluorescence at 490 nm to that at 520 nm. The IC50 for each compound was derived from fitting the data to the four-parameter logistic equation by Graphpad Prism software or Dotmatics. | B | 7.55 | pIC50 | 28 | nM | IC50 | US-11420968-B2. Bcl-2 inhibitors (2022) |
| ChEMBL | Inhibition of Bcl-2 G101V mutant (unknown origin) using (Ac-GQVGRQLAIIGDK (FITC) INR-amide) as substrate preincubated for 30 mins followed by substrate addition and measured after 60 mins by TR-FRET assay | B | 7.6 | pIC50 | 25 | nM | IC50 | J Med Chem (2024) 67: 7836-7858 [PMID:38695063] |
| ChEMBL | Inhibition of BCL-2 (unknown origin) | B | 7.64 | pIC50 | 23 | nM | IC50 | J Med Chem (2021) 64: 1362-1391 [PMID:33523672] |
| ChEMBL | Bcl-2 Competition Binding (Fluorescence Polarization) Assay: The fluorescence-labeled 23 amino acid peptide BH3 was purchased from CalBiochem (NLWAAQRYGRELRRMSDKFVD, SEQ ID NO: 1). An unbound Fluorescein labeled BH3 peptide emits random light with respect to the plane of polarization plane of excited light, resulting in a lower polarization degree (mP) value. When the peptide is bound to Bcl-2, the complex tumble slower and the emitted light can have a higher level of polarization, resulting in a higher mP value. This binding assay was performed in 96-well plate and with each assay contained 15 and 30 nM of labeled peptide and purified Bcl-2 protein (purchased from R&D Systems, Inc). The assay buffer contained 20 mM Hepes (pH 7.0), 50 mM KCl, 5 mM MgCl2, 20 mM Na2MoO4, 0.1 mg/ml Bovine Gamma Globulin and 0.01% NP40. Compounds were diluted in DMSO and added to the final assay with concentration range from 20 uM to 2 nM. The polarization degree (mP) value was determined by BioTek Synergy II with background subtraction after 3 hours of incubation at room temperature. IC50 was calculated using Prism software with sigmoidal dose-response curve fitting. ABT-737 was used as reference compound. Such assays, carried out with a range of doses of test compounds, allow the determination of an approximate IC50 value. Although the inhibitory properties of the compounds of the present invention vary with structural change as expected, the activity generally exhibited by these agents is in the range of IC50=0.1-1000 nM. | B | 8 | pIC50 | <10 | nM | IC50 | US-10377755-B2. BCL-2 inhibitors (2019) |
| ChEMBL | Homogeneous Time Resolved Fluorescence (HTRF) Assay: Table 2: Binding to Bcl-2 proteins Bcl-2, and Bcl-XL was also assessed using an HTRF assay. Background: FAM-Bak/Bad binds to surface pocket of the Bcl-2 protein family. This binding can be monitored by HTRF signals between anti-GST-Tb and FAM-peptide using GST-tagged Bcl proteins. Assay conditions: Bcl-2: 4 nM Bcl-2, 100 nM FAM-Bak peptide, Bcl-XL: 3 nM Bcl-XL, 40 nM FAM-Bad peptide in 20 mM K Phosphate, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.005% Triton X-100 and 1% DMSO (final). Assay procedure: Compounds were tested in 10-dose IC50 mode, in singlicate, with 3-fold serial dilution starting at 10 ÎĽM or 1 ÎĽM. Compound stock solutions were added to protein solution using Acoustic technology. | B | 8 | pIC50 | <10 | nM | IC50 | US-11344546-B2. Benzamide compounds (2022) |
| ChEMBL | Homogeneous Time Resolved Fluorescence (HTRF) Assay: Table 2: Binding to Bcl-2 proteins Bcl-2, and Bcl-XL was also assessed using an HTRF assay. Background: FAM-Bak/Bad binds to surface pocket of the Bcl-2 protein family. This binding can be monitored by HTRF signals between anti-GST-Tb and FAM-peptide using GST-tagged Bcl proteins. Assay conditions: Bcl-2: 4 nM Bcl-2, 100 nM FAM-Bak peptide, Bcl-XL: 3 nM Bcl-XL, 40 nM FAM-Bad peptide in 20 mM K Phosphate, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.005% Triton X-100 and 1% DMSO (final). Assay procedure: Compounds were tested in 10-dose IC50 mode, in singlicate, with 3-fold serial dilution starting at 10 ÎĽM or 1 ÎĽM. Compound stock solutions were added to protein solution using Acoustic technology. The compounds were then incubated with protein for 10 min at room temperature. The respective FAM labeled peptide was added and incubated for another 10 min and then anti-GST-Tb was added. After 60 min at rt, the HTRF fluorescence signal ratio was measured. | B | 8 | pIC50 | <10 | nM | IC50 | US-11318134-B2. Benzamide compounds (2022) |
| ChEMBL | Inhibition of FAM-labelled Bax binding to Bcl2 (unknown origin) after 30 mins by fluorescence polarization assay | B | 8.13 | pIC50 | 7.4 | nM | IC50 | Eur J Med Chem (2018) 159: 149-165 [PMID:30278333] |
| ChEMBL | Displacement of Bax-derived peptide from Bcl-2 (unknown origin) by fluorescence polarization assay | B | 8.13 | pIC50 | 7.39 | nM | IC50 | Bioorg Med Chem (2018) 26: 443-454 [PMID:29229225] |
| ChEMBL | Bcl-2/Bcl-X TR-FRET Assay: Compounds disclosed herein were tested for blocking of Bcl-2/Bcl-X protein with its ligand in an assay based on Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) methodology. Recombinant human 0.05 nM Bcl-2/0.03 nM Bcl-X protein was pre-incubated with a serial dilution of compounds disclosed herein (maximum concentration is 0.1 ÎĽM for Bcl-2 assay, and 10 ÎĽM for Bcl-xl assay, 3-fold serially diluted, 10 points; or maximum concentration is 0.02 ÎĽM for Bcl-2 assay, and 2 ÎĽM for Bcl-xl assay, 3-fold serially diluted, 10 points) at room temperature for 0.5 hour in an assay buffer containing 20 mM potassium phosphate buffer, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.05% Tween-20, 0.01% BSA. Then the FITC labeled Bak peptide Ac-GQVGRQLAIIGDK(FITC)INR-amide (0.5 nM for Bcl-2, 0.3 nM for Bcl-xl) and MAb Anti 6His Tb cryptate Gold were added to plate and further incubated at room temperature for 1 hour. The TR-FRET signals (337 nm-520 nm-490 nm) were read on BMG PHERAstar FSX instrument. The inhibition percentage of Bcl-2/Bcl-X interaction with its ligand in presence of increasing concentrations of compounds was calculated based on the TR-FRET signals. The IC50 for each compound was derived from fitting the data to the four-parameter logistic equation by Graphpad Prism software. To improve the assay sensitivity and test more potent compounds in the present application, the bcl-2 concentration was reduced in method B. | B | 8.64 | pIC50 | 2.3 | nM | IC50 | US-11420968-B2. Bcl-2 inhibitors (2022) |
| ChEMBL | Bcl-2-G101V Biochemical Assay : Selected compounds disclosed herein were tested for blocking of Bcl-2-G101 protein with its ligand in an assay based on time-resolved fluorescence resonance energy transfer methodology. 0.05 nM of Recombinant human Bcl-2-G101V protein was pre-incubated with a serial dilution of compounds disclosed herein (maximum concentration is 10 ÎĽM, 4-fold serially diluted, 10 points; or maximum concentration is 1 uM, 3-fold serially diluted, 10 points) at room temperature for 0.5 hour in an assay buffer containing 20 mM potassium phosphate buffer, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.05% Tween-20, 0.01% BSA. Then 5 nM of the FITC labeled Bak peptide Ac-GQVGRQLAIIGDK(FITC)INR-amide and Mab Anti-6His Tb cryptate Gold was added to plate and further incubated at room temperature for 1 hour. The TR-FRET signals (ex337 nm, em490 nm/520 nm) were read on BMG PHERAstar FSX instrument. The inhibition percentage of Bcl-2-G101V interaction with its ligand in presence of increasing concentrations of compounds was calculated based on the ratio of fluorescence at 490 nm to that at 520 nm. The IC50 for each compound was derived from fitting the data to the four-parameter logistic equation by Graphpad Prism software or Dotmatics. | B | 8.92 | pIC50 | 1.2 | nM | IC50 | US-11420968-B2. Bcl-2 inhibitors (2022) |
| ChEMBL | Displacement of Bak derived peptide from Bcl-2 (unknown origin) measured after 15 mins by microplate reader assay | B | 9 | pIC50 | 1 | nM | IC50 | Bioorg Med Chem (2021) 47: 116350-116350 [PMID:34536651] |
| ChEMBL | Inhibition of FAM-Bid binding to human BCL2 expressed in Escherichia coli BL21 after 30 mins by fluorescence polarization assay | B | 9 | pIC50 | <1 | nM | IC50 | ACS Med Chem Lett (2016) 7: 1185-1190 [PMID:27994761] |
| ChEMBL | Bcl-2/Bcl-X TR-FRET Assay: Compounds disclosed herein were tested for blocking of Bcl-2/Bcl-X protein with its ligand in an assay based on Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) methodology. Recombinant human 0.05 nM Bcl-2/0.03 nM Bcl-X protein was pre-incubated with a serial dilution of compounds disclosed herein (maximum concentration is 0.1 ÎĽM for Bcl-2 assay, and 10 ÎĽM for Bcl-xl assay, 3-fold serially diluted, 10 points; or maximum concentration is 0.02 ÎĽM for Bcl-2 assay, and 2 ÎĽM for Bcl-xl assay, 3-fold serially diluted, 10 points) at room temperature for 0.5 hour in an assay buffer containing 20 mM potassium phosphate buffer, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.05% Tween-20, 0.01% BSA. Then the FITC labeled Bak peptide Ac-GQVGRQLAIIGDK(FITC)INR-amide (0.5 nM for Bcl-2, 0.3 nM for Bcl-xl) and MAb Anti 6His Tb cryptate Gold were added to plate and further incubated at room temperature for 1 hour. The TR-FRET signals (337 nm-520 nm-490 nm) were read on BMG PHERAstar FSX instrument. The inhibition percentage of Bcl-2/Bcl-X interaction with its ligand in presence of increasing concentrations of compounds was calculated based on the TR-FRET signals. The IC50 for each compound was derived from fitting the data to the four-parameter logistic equation by Graphpad Prism software. To improve the assay sensitivity and test more potent compounds in the present application, the bcl-2 concentration was reduced in method B. | B | 9.47 | pIC50 | 0.34 | nM | IC50 | US-11420968-B2. Bcl-2 inhibitors (2022) |
| ChEMBL | Inhibition of Bcl-2 (unknown origin) using (Ac-GQVGRQLAIIGDK (FITC) INR-amide) as substrate preincubated for 30 mins followed by substrate addition and measured after 60 mins by TR-FRET assay | B | 9.62 | pIC50 | 0.24 | nM | IC50 | J Med Chem (2024) 67: 7836-7858 [PMID:38695063] |
| ChEMBL | Inhibition of BCL-2 (unknown origin) by fluorescence polarization assay | B | 8.52 | pEC50 | 3 | nM | EC50 | Eur J Med Chem (2019) 167: 76-95 [PMID:30769242] |
| Aspartyl/asparaginyl beta-hydroxylase in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4680030] [UniProtKB: Q12797] | ||||||||
| ChEMBL | Inhibition of N-His6-tagged human AspH (315-755) expressed in Escherichia coli BL21 (DE3) using high 10 uM hFX-CP as substrate mixture with 10 uM 2OG, 100 uM L-ascorbic acid and 2 uM FAS incubated for 35 mins by MS analysis | B | 5.8 | pIC50 | 1570 | nM | IC50 | Bioorg Med Chem (2020) 28: 115675-115675 [PMID:33069066] |
| ChEMBL | Inhibition of N-His6-tagged human AspH (315-755) expressed in Escherichia coli BL21 (DE3) using 1 uM hFX-CP as substrate mixture with high 200 uM 2OG, 100 uM L-ascorbic acid and 2 uM FAS incubated for 35 mins by MS analysis | B | 5.82 | pIC50 | 1520 | nM | IC50 | Bioorg Med Chem (2020) 28: 115675-115675 [PMID:33069066] |
| ChEMBL | Inhibition of N-His6-tagged human AspH (315-755) expressed in Escherichia coli BL21 (DE3) using 1 uM hFX-CP as substrate mixture with 3 uM 2OG, 100 uM L-ascorbic acid and 2 uM FAS incubated for 35 mins by MS analysis | B | 5.85 | pIC50 | 1400 | nM | IC50 | Bioorg Med Chem (2020) 28: 115675-115675 [PMID:33069066] |
| ChEMBL | Inhibition of N-His6-tagged human AspH (315-755) expressed in Escherichia coli BL21 (DE3) using 1 uM hFX-CP as substrate mixture with 3 uM 2OG, 100 uM L-ascorbic acid and high 20 uM FAS incubated for 35 mins by MS analysis | B | 5.89 | pIC50 | 1290 | nM | IC50 | Bioorg Med Chem (2020) 28: 115675-115675 [PMID:33069066] |
| Bcl2-associated agonist of cell death in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3817] [UniProtKB: Q92934] | ||||||||
| ChEMBL | Binding Assay: Table 1: Binding to Bcl-2 proteins Bcl-2, and Bcl-XL was assessed using the Bcl2scan™ platform: T7 phage strains displaying BCL2 proteins were grown in parallel in 24-well blocks in an E. coli host derived from the BL21 strain. E. coli were grown to log-phase and infected with T7 phage from a frozen stock (multiplicity of infection=0.4) and incubated with shaking at 32° C. until lysis (90-150 minutes). The lysates were centrifuged (5,000×g) and filtered (0.2 μm) to remove cell debris. Streptavidin-coated magnetic beads were treated with biotinylated BIM peptide ligand for 30 minutes at room temperature to generate affinity resins for BCL2 assays. The liganded beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific phage binding. Binding reactions were assembled by combining BCL2 proteins, liganded affinity beads, and test compounds in 1× binding buffer (20% SeaBlock, 0.17×PBS, 0.05% Tween 20, 6 mM DTT). Test compounds were prepared as 100× stocks in 100% DMSO. Kds were determined using an 11-point 3-fold compound dilution series with one DMSO control point. All compounds for Kd measurements were distributed by acoustic transfer in 100% DMSO. The compounds were then diluted directly into the assays such that the final concentration of DMSO was 0.9%. All reactions performed in polypropylene 384-well plates. Each was a final volume of 0.02 ml. The assay plates were incubated at room temperature with shaking for 1 hour and the affinity beads were washed with wash buffer (lx PBS, 0.05% Tween 20). | B | 7.26 | pKd | 55 | nM | Kd | US-11344546-B2. Benzamide compounds (2022) |
| ChEMBL | Binding affinity to Bcl-2 (unknown origin) by fluorescence polarization competition assay | B | 4.8 | pKi | 16000 | nM | Ki | Bioorg Med Chem Lett (2021) 47: 128215-128215 [PMID:34153472] |
| ChEMBL | Homogeneous Time Resolved Fluorescence (HTRF) Assay: Table 2: Binding to Bcl-2 proteins Bcl-2, and Bcl-XL was also assessed using an HTRF assay. Background: FAM-Bak/Bad binds to surface pocket of the Bcl-2 protein family. This binding can be monitored by HTRF signals between anti-GST-Tb and FAM-peptide using GST-tagged Bcl proteins. Assay conditions: Bcl-2: 4 nM Bcl-2, 100 nM FAM-Bak peptide, Bcl-XL: 3 nM Bcl-XL, 40 nM FAM-Bad peptide in 20 mM K Phosphate, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.005% Triton X-100 and 1% DMSO (final). Assay procedure: Compounds were tested in 10-dose IC50 mode, in singlicate, with 3-fold serial dilution starting at 10 ÎĽM or 1 ÎĽM. Compound stock solutions were added to protein solution using Acoustic technology. | B | 7.26 | pIC50 | 55 | nM | IC50 | US-11344546-B2. Benzamide compounds (2022) |
| Bcl-2-like 1/Bcl-2-like protein 1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4625] [GtoPdb: 2845] [UniProtKB: Q07817] | ||||||||
| ChEMBL | Binding Assay: Table 1: Binding to Bcl-2 proteins Bcl-2, and Bcl-XL was assessed using the Bcl2scan™ platform: T7 phage strains displaying BCL2 proteins were grown in parallel in 24-well blocks in an E. coli host derived from the BL21 strain. E. coli were grown to log-phase and infected with T7 phage from a frozen stock (multiplicity of infection=0.4) and incubated with shaking at 32° C. until lysis (90-150 minutes). The lysates were centrifuged (5,000×g) and filtered (0.2 μm) to remove cell debris. Streptavidin-coated magnetic beads were treated with biotinylated BIM peptide ligand for 30 minutes at room temperature to generate affinity resins for BCL2 assays. The liganded beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific phage binding. Binding reactions were assembled by combining BCL2 proteins, liganded affinity beads, and test compounds in 1× binding buffer (20% SeaBlock, 0.17×PBS, 0.05% Tween 20, 6 mM DTT). Test compounds were prepared as 100× stocks in 100% DMSO. Kds were determined using an 11-point 3-fold compound dilution series with one DMSO control point. All compounds for Kd measurements were distributed by acoustic transfer in 100% DMSO. The compounds were then diluted directly into the assays such that the final concentration of DMSO was 0.9%. All reactions performed in polypropylene 384-well plates. Each was a final volume of 0.02 ml. The assay plates were incubated at room temperature with shaking for 1 hour and the affinity beads were washed with wash buffer (lx PBS, 0.05% Tween 20). The beads were then re-suspended in elution buffer (lx PBS, 0.05% Tween 20, 2 μM non-biotinylated affinity ligand) and incubated at room temperature with shaking for 30 minutes. | B | 7.26 | pKd | 55 | nM | Kd | US-11318134-B2. Benzamide compounds (2022) |
| ChEMBL | Bcl-2 Protein Family Binding Assay: Binding to Bcl-2 proteins Bcl-2, and Bcl-XL was assessed using the Bcl2scan™ platform: T7 phage strains displaying BCL2 proteins were grown in parallel in 24-well blocks in an E. coli host derived from the BL21 strain. E. coliwere grown to log-phase and infected with T7 phage from a frozen stock (multiplicity of infection=0.4) and incubated with shaking at 32° C. until lysis (90-150 minutes). The lysates were centrifuged (5,000×g) and filtered (0.2 μm) to remove cell debris. Streptavidin-coated magnetic beads were treated with biotinylated BIM peptide ligand for 30 minutes at room temperature to generate affinity resins for BCL2 assays. The liganded beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific phage binding. Binding reactions were assembled by combining BCL2 proteins, liganded affinity beads, and test compounds in 1× binding buffer (20% SeaBlock, 0.17×PBS, 0.05% Tween 20, 6 mM DTT). Test compounds were prepared as 100× stocks in 100% DMSO. Kds were determined using an 11-point 3-fold compound dilution series with one DMSO control point. All compounds for Kd measurements were distributed by acoustic transfer in 100% DMSO. The compounds were then diluted directly into the assays such that the final concentration of DMSO was 0.9%. All reactions performed in polypropylene 384-well plates. Each was a final volume of 0.02 ml. The assay plates were incubated at room temperature with shaking for 1 hour and the affinity beads were washed with wash buffer (lx PBS, 0.05% Tween 20). The beads were then re-suspended in elution buffer (lx PBS, 0.05% Tween 20, 2 μM non-biotinylated affinity ligand) and incubated at room temperature with shaking for 30 minutes. | B | 7.26 | pKd | 55 | nM | Kd | US-11590126-B2. Benzamide compounds (2023) |
| ChEMBL | Binding affinity to Bcl-XL (unknown origin) by FRET assay | B | 7.32 | pKi | 48 | nM | Ki | Bioorg Med Chem Lett (2016) 26: 2105-2114 [PMID:26988306] |
| ChEMBL | Binding affinity to His-tagged Bcl-XL (unknown origin) incubated for 30 mins by TR-FRET assay | B | 7.32 | pKi | 48 | nM | Ki | Medchemcomm (2016) 7: 778-787 |
| ChEMBL | Binding affinity to Bcl-xL (unknown origin) by TR-FRET assay | B | 7.32 | pKi | 48 | nM | Ki | J Med Chem (2017) 60: 821-838 [PMID:27749061] |
| ChEMBL | Inhibition of Bcl-xL (unknown origin) by TR-FRET assay | B | 7.32 | pKi | 48 | nM | Ki | Eur J Med Chem (2018) 146: 471-482 [PMID:29407973] |
| ChEMBL | Inhibition of Bcl-xL (unknown origin) | B | 7.32 | pKi | 48 | nM | Ki | J Med Chem (2018) 61: 2636-2651 [PMID:28926247] |
| ChEMBL | Binding affinity to BCL-xL (unknown origin) assessed as inhibition constant by fluorescence polarization binding affinity assay | B | 7.32 | pKi | 48 | nM | Ki | J Med Chem (2024) 67: 10795-10830 [PMID:38913996] |
| ChEMBL | Binding affinity to Bcl-xl (unknown origin) assessed as dissociation constant incubated for 1 hrs by TR-FRET assay | B | 7.32 | pKi | 48 | nM | Ki | Sci Transl Med (2015) 7: null-null [PMID:25787766] |
| ChEMBL | Displacement of fluorescent-labeled BID-BH3 peptide from human BCL-xL expressed in Escherichia coli BL21 (DE3) assessed as inhibition constant by fluorescence polarization-based competitive binding assay | B | 7.32 | pKi | 48 | nM | Ki | Eur J Med Chem (2022) 236: 114327-114327 [PMID:35385805] |
| ChEMBL | Binding affinity to Bcl-xL (unknown origin) assessed as inhibition constant by TR-FRET assay | B | 7.32 | pKi | 48 | nM | Ki | Eur J Med Chem (2020) 201: 112446-112446 [PMID:32563811] |
| GtoPdb | - | - | 7.32 | pKi | 48 | nM | Ki | Nat Med (2013) 19: 202-8 [PMID:23291630] |
| ChEMBL | Binding affinity to Bc1-xL (unknown origin) by fluorescence polarization competition assay | B | 9 | pKi | 1 | nM | Ki | Bioorg Med Chem Lett (2021) 47: 128215-128215 [PMID:34153472] |
| ChEMBL | Binding affinity to full length human Bcl-xl expressed in Escherichia coli BL21 (DE3) assessed as inhibition constant preincubated for 30 mins followed by 5-FAM-QEDIIIINIARHLAQVGDSMD-RSIPPG tracer addition and measured after 20 mins by fluorescence polarization assay | B | 9 | pKi | <1 | nM | Ki | J Med Chem (2021) 64: 10260-10285 [PMID:34228434] |
| ChEMBL | Binding affinity to Bcl-xL (unknown origin) pre-incubated for 30 mins followed by addition FAM-peptide further incubated for 20 mins by fluorescence polarization-based binding assay | B | 4 | pIC50 | >100000 | nM | IC50 | J Med Chem (2024) 67: 13925-13958 [PMID:39121336] |
| ChEMBL | Bcl-2/Bcl-X TR-FRET Assay: Compounds disclosed herein were tested for blocking of Bcl-2/Bcl-X protein with its ligand in an assay based on Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) methodology. Recombinant human 0.05 nM Bcl-2/0.03 nM Bcl-X protein was pre-incubated with a serial dilution of compounds disclosed herein (maximum concentration is 0.1 ÎĽM for Bcl-2 assay, and 10 ÎĽM for Bcl-xl assay, 3-fold serially diluted, 10 points; or maximum concentration is 0.02 ÎĽM for Bcl-2 assay, and 2 ÎĽM for Bcl-xl assay, 3-fold serially diluted, 10 points) at room temperature for 0.5 hour in an assay buffer containing 20 mM potassium phosphate buffer, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.05% Tween-20, 0.01% BSA. Then the FITC labeled Bak peptide Ac-GQVGRQLAIIGDK(FITC)INR-amide (0.5 nM for Bcl-2, 0.3 nM for Bcl-xl) and MAb Anti 6His Tb cryptate Gold were added to plate and further incubated at room temperature for 1 hour. The TR-FRET signals (337 nm-520 nm-490 nm) were read on BMG PHERAstar FSX instrument. The inhibition percentage of Bcl-2/Bcl-X interaction with its ligand in presence of increasing concentrations of compounds was calculated based on the TR-FRET signals. The IC50 for each compound was derived from fitting the data to the four-parameter logistic equation by Graphpad Prism software. To improve the assay sensitivity and test more potent compounds in the present application, the bcl-2 concentration was reduced in method B. | B | 6.72 | pIC50 | 190 | nM | IC50 | US-11420968-B2. Bcl-2 inhibitors (2022) |
| ChEMBL | Displacement of Bak derived peptide from Bcl-xL (unknown origin) measured after 15 mins by microplate reader assay | B | 6.8 | pIC50 | 157 | nM | IC50 | Bioorg Med Chem (2021) 47: 116350-116350 [PMID:34536651] |
| ChEMBL | Inhibition of Bcl-Xl (unknown origin) | B | 7.19 | pIC50 | 65 | nM | IC50 | J Med Chem (2024) 67: 7836-7858 [PMID:38695063] |
| ChEMBL | Homogeneous Time Resolved Fluorescence (HTRF) Assay: Table 2: Binding to Bcl-2 proteins Bcl-2, and Bcl-XL was also assessed using an HTRF assay. Background: FAM-Bak/Bad binds to surface pocket of the Bcl-2 protein family. This binding can be monitored by HTRF signals between anti-GST-Tb and FAM-peptide using GST-tagged Bcl proteins. Assay conditions: Bcl-2: 4 nM Bcl-2, 100 nM FAM-Bak peptide, Bcl-XL: 3 nM Bcl-XL, 40 nM FAM-Bad peptide in 20 mM K Phosphate, pH 7.5, 50 mM NaCl, 1 mM EDTA, 0.005% Triton X-100 and 1% DMSO (final). Assay procedure: Compounds were tested in 10-dose IC50 mode, in singlicate, with 3-fold serial dilution starting at 10 ÎĽM or 1 ÎĽM. Compound stock solutions were added to protein solution using Acoustic technology. The compounds were then incubated with protein for 10 min at room temperature. The respective FAM labeled peptide was added and incubated for another 10 min and then anti-GST-Tb was added. After 60 min at rt, the HTRF fluorescence signal ratio was measured. | B | 7.26 | pIC50 | 55 | nM | IC50 | US-11318134-B2. Benzamide compounds (2022) |
| Bcl-2-like 2/Bcl-2-like protein 2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4677] [GtoPdb: 2846] [UniProtKB: Q92843] | ||||||||
| GtoPdb | - | - | 6.61 | pKi | 245 | nM | Ki | Nat Med (2013) 19: 202-8 [PMID:23291630] |
| ChEMBL | Binding affinity to Bcl-w (unknown origin) by TR-FRET assay | B | 6.61 | pKi | 245 | nM | Ki | J Med Chem (2017) 60: 821-838 [PMID:27749061] |
| ChEMBL | Displacement of fluorescent-labeled BID-BH3 peptide from human BCL-W expressed in Escherichia coli BL21 (DE3) assessed as inhibition constant by fluorescence polarization-based competitive binding assay | B | 6.61 | pKi | 245 | nM | Ki | Eur J Med Chem (2022) 236: 114327-114327 [PMID:35385805] |
| ChEMBL | Binding affinity to Bcl-w (unknown origin) assessed as inhibition constant by TR-FRET assay | B | 6.61 | pKi | 245 | nM | Ki | Eur J Med Chem (2020) 201: 112446-112446 [PMID:32563811] |
| ChEMBL | Binding affinity to BCL-2 (unknown origin) assessed as inhibition constant by fluorescence polarization binding affinity assay | B | 6.61 | pKi | 245 | nM | Ki | J Med Chem (2024) 67: 10795-10830 [PMID:38913996] |
| ChEMBL | Binding affinity to BCL-W (unknown origin) assessed as dissociation constant incubated for 1 hrs by TR-FRET assay | B | 6.61 | pKi | 245 | nM | Ki | Sci Transl Med (2015) 7: null-null [PMID:25787766] |
| CYP2C9/Cytochrome P450 2C9 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3397] [GtoPdb: 1326] [UniProtKB: P11712] | ||||||||
| ChEMBL | CYP 2C9 inhibition: The five isoform-selective probe substrate (in a cocktail manner) was used as a measure of activity for the individual cytochrome P450 (CYPs) in a pool of human liver microsomes, i.e., phenacetin for CYP1A2, diclofenac for CYP2C9, S-Mephenyloin for CYP2C19, dextromethorphan for CYP2D6, midazolam for CYP3A. Test compounds, at 7 concentration levels including zero, were incubated in human liver microsomes (HLM) together with the 5 probe substrate (in a cocktail manner). IC50 was determined by monitoring the reduction of the CYP activity as a function of test compound concentration and quantified by product formation using LC-MS/MS. Ketoconazole for CYP3A was included as quality control. All incubations were performed in singlet. | B | 5.75 | pIC50 | 1770 | nM | IC50 | US-11420968-B2. Bcl-2 inhibitors (2022) |
| nuclear receptor binding SET domain protein 1/Histone-lysine N-methyltransferase, H3 lysine-36 specific in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3588738] [GtoPdb: 2696] [UniProtKB: Q96L73] | ||||||||
| ChEMBL | Inhibition of N-terminal polyhistidine-tagged recombinant human NSD1 (1538 to 2696 residues) expressed in baculovirus infected insect cell using SAM as substrate preincubated for 20 mins followed by substrate addition measured after 1 hrs by hotspot assay | B | 5.62 | pIC50 | 2400 | nM | IC50 | Eur J Med Chem (2023) 256: 115440-115440 [PMID:37182335] |
| nuclear receptor binding SET domain protein 2/Histone-lysine N-methyltransferase NSD2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3108645] [GtoPdb: 3220] [UniProtKB: O96028] | ||||||||
| ChEMBL | Inhibition of N-terminal polyhistidine-tagged human NSD2 expressed in baculovirus infected insect cell using SAM as substrate preincubated for 20 mins followed by substrate addition measured after 1 hrs by hotspot assay | B | 5.77 | pIC50 | 1700 | nM | IC50 | Eur J Med Chem (2023) 256: 115440-115440 [PMID:37182335] |
| Histone-lysine N-methyltransferase NSD3 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3108646] [UniProtKB: Q9BZ95] | ||||||||
| ChEMBL | Inhibition of N-terminal GST-fused recombinant human NSD3 (1021 to 1322 residues) expressed in Escherichia coli using SAM as substrate preincubated for 20 mins followed by substrate addition measured after 1 hrs by hotspot assay | B | 5.89 | pIC50 | 1300 | nM | IC50 | Eur J Med Chem (2023) 256: 115440-115440 [PMID:37182335] |
| MCL1 apoptosis regulator, BCL2 family member/Induced myeloid leukemia cell differentiation protein Mcl-1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4361] [GtoPdb: 2847] [UniProtKB: Q07820] | ||||||||
| ChEMBL | Inhibition of FAM-Bid binding to human MCL1 expressed in Escherichia coli BL21 after 30 mins by fluorescence polarization assay | B | 6.11 | pKi | >780 | nM | Ki | ACS Med Chem Lett (2016) 7: 1185-1190 [PMID:27994761] |
| GtoPdb | - | - | 6.35 | pKi | >444 | nM | Ki | Nat Med (2013) 19: 202-8 [PMID:23291630] |
| ChEMBL | Binding affinity to Mcl-1 (unknown origin) by TR-FRET assay | B | 6.35 | pKi | >444 | nM | Ki | J Med Chem (2017) 60: 821-838 [PMID:27749061] |
| ChEMBL | Inhibition of Mcl-1 (unknown origin) by TR-FRET assay | B | 6.35 | pKi | >444 | nM | Ki | Eur J Med Chem (2018) 146: 471-482 [PMID:29407973] |
| ChEMBL | Displacement of fluorescent-labeled BID-BH3 peptide from His-tagged human MCL-1 expressed in Escherichia coli BL21 (DE3) assessed as inhibition constant by fluorescence polarization-based competitive binding assay | B | 6.35 | pKi | >444 | nM | Ki | Eur J Med Chem (2022) 236: 114327-114327 [PMID:35385805] |
| ChEMBL | Binding affinity to Mcl-1 (unknown origin) assessed as inhibition constant by TR-FRET assay | B | 6.35 | pKi | >444 | nM | Ki | Eur J Med Chem (2020) 201: 112446-112446 [PMID:32563811] |
| ChEMBL | Binding affinity to MCL-1 (unknown origin) assessed as inhibition constant by time-resolved fluorescence resonance energy transfer binding affinity assays | B | 6.35 | pKi | >444 | nM | Ki | J Med Chem (2024) 67: 10795-10830 [PMID:38913996] |
| ChEMBL | Binding affinity to MCL1 (unknown origin) incubated for 30 mins by TR-FRET assay | B | 6.35 | pKi | >444 | nM | Ki | Medchemcomm (2016) 7: 778-787 |
| ChEMBL | Binding affinity to MCL-1 (unknown origin) assessed as dissociation constant incubated for 1 hrs by TR-FRET assay | B | 6.35 | pKi | >444 | nM | Ki | Sci Transl Med (2015) 7: null-null [PMID:25787766] |
| ChEMBL | Inhibition of FAM-Bid binding to human MCL1 expressed in Escherichia coli BL21 after 30 mins by fluorescence polarization assay | B | 5.4 | pIC50 | >4000 | nM | IC50 | ACS Med Chem Lett (2016) 7: 1185-1190 [PMID:27994761] |
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]
