#	Aladdin ID	Assay Type	Description	Organism	Compounds	Reference	BAO Format	Source
1.	ALA1273510	B	Binding affinity Bcl-xL at 200 uM by one-dimensional 1H nuclear magnetic resonance spectroscopy		17	ALA1269007	single protein format	Scientific Literature
2.	ALA1273511	F	Cytotoxicity against human PC3 cells expressing high level of Bcl-xL after 72 hrs by ATP-LITE assay	Homo sapiens	17	ALA1269007	cell-based format	Scientific Literature
3.	ALA1273512	F	Cytotoxicity against human H460 cells expressing high level of Bcl-2 after 72 hrs by ATP-LITE assay	Homo sapiens	17	ALA1269007	cell-based format	Scientific Literature
4.	ALA1273513	F	Cytotoxicity against human H1299 cells expressing high level of Mcl-1 after 72 hrs by ATP-LITE assay	Homo sapiens	17	ALA1269007	cell-based format	Scientific Literature
5.	ALA1273514	F	Cytotoxicity against human BP3 cells expressing high level of Bfl-1 and Mcl-1 after 72 hrs by annexin V-FITC and propidium iodide staining	Homo sapiens	17	ALA1269007	cell-based format	Scientific Literature
6.	ALA1273515	F	Cytotoxicity against human RS4:11 cells expressing high level of Bcl-2 and Bcl-xL after 72 hrs by annexin V-FITC and propidium iodide staining	Homo sapiens	17	ALA1269007	cell-based format	Scientific Literature
7.	ALA1273516	B	Displacement of F-BakBH3 from Bcl-xL after 30 mins by fluorescence polarization assay		12	ALA1269007	single protein format	Scientific Literature
8.	ALA1273517	B	Displacement of F-BakBH3 from Bcl-2 after 30 mins by fluorescence polarization assay		12	ALA1269007	single protein format	Scientific Literature
9.	ALA1273518	B	Displacement of F-BakBH3 from Bfl-1 after 30 mins by fluorescence polarization assay		12	ALA1269007	single protein format	Scientific Literature
10.	ALA1273519	B	Displacement of F-BakBH3 from Mcl-1 after 30 mins by fluorescence polarization assay		12	ALA1269007	single protein format	Scientific Literature
11.	ALA1273520	B	Binding affinity Bcl-xL by isothermal titration calorimetry assay		12	ALA1269007	single protein format	Scientific Literature
12.	ALA1273521	A	Metabolic stability of the compound in rat plasma after 40 mins	Rattus norvegicus	8	ALA1269007	cell-free format	Scientific Literature
13.	ALA1273522	A	Metabolic stability of the compound in rat microsomes after 40 mins	Rattus norvegicus	8	ALA1269007	microsome format	Scientific Literature
14.	ALA1273523	A	Permeability of the compound by PAMPA		8	ALA1269007	assay format	Scientific Literature
15.	ALA1273526	F	Reduction of spleen weight in ip dosed human Bcl2 expressing B6 transgenic BALB/c mouse administered as single dose after 24 hrs	Mus musculus	17	ALA1269007	organism-based format	Scientific Literature
16.	ALA1273528	F	Reduction of spleen weight in human Bcl2 expressing B6 transgenic BALB/c mouse at 60 umol/kg, ip administered as single dose after 24 hrs	Mus musculus	3	ALA1269007	organism-based format	Scientific Literature
17.	ALA1273529	T	Toxicity in human Bcl2 expressing B6 transgenic BALB/c mouse up to 120 umol/kg, ip administered as single dose after 24 hrs	Mus musculus	3	ALA1269007	organism-based format	Scientific Literature
18.	ALA3705613	B	ITC Assay: The synthesized compounds of Formula I may be screened by one-dimensional 1H nuclear magnetic resonance spectroscopy (1D-1H NMR) binding assays against Bcl-XL. Active compounds in 1D-1H NMR binding assays are then selected and evaluated in the Isothermal Titration Calorimetry assays (ITC), cell viability assays and competitive fluorescence polarization assays (FPA). A group of compounds of Formula I display high binding affinity for Bcl-XL in these assays. The most potent compounds induce significant chemical shift changes in the active site methyl groups (region between 0.38 and 0.42 ppm) in the one-dimensional 1H-NMR spectra of Bcl-XL and also have an IC50 value in the FP displacement assays, which is more effective than Apogossypol. To confirm results of the NMR binding data and the FP assays, binding affinity of the compounds of Formula I for Bcl-XL using ITC assay were tested.	Homo sapiens	4	ALA3638406	single protein format	BindingDB Database
19.	ALA3705614	B	Fluorescence Polarization Assay: A Bak BH3 peptide (F-BakBH3) (GQVGRQLAIIGDDINR (SEQ ID NO:1)) was labeled at the N-terminus with fluorescein isothiocyanate (FITC) (Molecular Probes) and purified by HPLC. For competitive binding assays, 100 nM GST-BCL-XL DTM protein was preincubated with the tested compound at varying concentrations in 47.5 uL PBS (pH=7.4) in 96-well black plates at room temperature for 10 min, then 2.5 uL of 100 nM FITC-labeled Bak BH3 peptide was added to produce a final volume of 50 uL. The wild-type and mutant Bak BH3 peptides were included in each assay plate as positive and negative controls, respectively. After 30 min incubation at room temperature, the polarization values in millipolarization units were measured at excitation/emission wavelengths of 480/535 nm with a multilabel plate reader (PerkinElmer). IC50 was determined by fitting the experimental data to a sigmoidal dose-response nonlinear regression model.	Homo sapiens	12	ALA3638406	single protein format	BindingDB Database
20.	ALA3707868	B	Fluorescence Polarization Assay: A Bak BH3 peptide (F-BakBH3) (GQVGRQLAIIGDDINR (SEQ ID NO:1)) was labeled at the N-terminus with fluorescein isothiocyanate (FITC) (Molecular Probes) and purified by HPLC. For competitive binding assays, 100 nM GST-BCL-XL DTM protein was preincubated with the tested compound at varying concentrations in 47.5 uL PBS (pH=7.4) in 96-well black plates at room temperature for 10 min, then 2.5 uL of 100 nM FITC-labeled Bak BH3 peptide was added to produce a final volume of 50 uL. The wild-type and mutant Bak BH3 peptides were included in each assay plate as positive and negative controls, respectively. After 30 min incubation at room temperature, the polarization values in millipolarization units were measured at excitation/emission wavelengths of 480/535 nm with a multilabel plate reader (PerkinElmer). IC50 was determined by fitting the experimental data to a sigmoidal dose-response nonlinear regression model.	Homo sapiens	12	ALA3638406	single protein format	BindingDB Database