| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
Bax (Kd = 15.0 μM)
BAI1 targets BAX (Bcl-2-associated X protein). The dissociation constant (Kd) for BAX binding is 15.0 ± 4 μM as determined by microscale thermophoresis. IC50 for inhibition of tBID-induced BAX-mediated membrane permeabilization is 3.3 μM. IC50 for inhibition of tBID-induced BAX membrane translocation is 5 ± 1 μM; for BIM SAHB-induced translocation, IC50 = 2 ± 1 μM. IC50 for inhibition of TNFα/cycloheximide-mediated caspase 3/7 activation in wild-type MEFs is 1.8 μM [1]. |
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| ln Vitro |
BAX activation inhibitor 1 (BAI1) demonstrates inhibition of tBID-induced BAX-mediated membrane permeabilization in a dose dependent manner with an IC50 of 3.3 μM. At an original allosteric site, BAI1 binds to monomeric BAX. While BAI1 does not directly compete with the activator to inhibit conformational activation and oligomerization of BAX in solution, it does so through an allosteric mechanism that favors the inactive BAX. Additionally, BAI1 inhibited BAX membrane association and translocation brought about by tBID (IC50 = 5 ± 1 μΜ) or BIM SAHB (IC50 = 2 ± 1 μΜ) in a dose-dependent manner and with an IC50 that was comparable to BAX-mediated liposomal release. Only BAX-dependent cell death is inhibited by BAI1[1].
BAI1 directly binds to inactive BAX with a Kd of 15.0 ± 4 μM (MST). It inhibits tBID-induced BAX-mediated liposomal membrane permeabilization with IC50 = 3.3 μM. BAI1 inhibits tBID-induced BAX oligomerization and dimerization in the presence of liposomal membranes as detected by BMH cross-linking and SDS-PAGE. It inhibits tBID-induced BAX membrane translocation with IC50 = 5 ± 1 μM and BIM SAHB-induced translocation with IC50 = 2 ± 1 μM. In NMR-based activation assays, BAI1 (100 μM) strongly inhibits BIM SAHB-induced BAX conformational activation and oligomerization, preserving ~80% of monomeric BAX at 24 h and 50% at 12 days. BAI1 suppresses BIM SAHB-induced exposure of the BH3 domain of BAX in a pull-down assay with anti-BH3 domain antibody. BAI1 increases the thermal stability of BAX, raising its activation temperature by ~10°C (from ~50°C to ~60°C). Hydrogen-deuterium exchange mass spectrometry and paramagnetic relaxation enhancement (PRE) experiments show that BAI1 protects regions around the BAI-site and hydrophobic core residues of helix α5 [1]. |
| Enzyme Assay |
Binding of BAI1 to BAX was evaluated by ligand-detected 1D 1H-NMR, STD, CPMG experiments, and 2D 1H-15N HSQC NMR. For ligand-detected NMR, 100-150 μM BAI1 was incubated with or without unlabeled BAX (10-15 μM) in 50 mM potassium phosphate, 50 mM NaCl, pH 6.0, 10% D2O. STD experiments used off-resonance frequency of -2000 Hz and on-resonance frequency of 450 Hz with saturation time 0.5-2 s. CPMG experiments used relaxation delay of 200 ms. For HSQC, 15N-labeled BAX (50 μM) was titrated with up to 150 μM BAI1; spectra were acquired at 25°C on 600 MHz spectrometer, and chemical shift perturbations (CSPs) calculated. Microscale thermophoresis (MST): 100 nM labeled BAX was incubated with increasing concentrations of BAI1 (150-0.5 μM) in MST buffer (100 mM potassium phosphate, pH 7.4, 150 mM NaCl), sonicated for 10 s, loaded into capillaries, and analyzed with Monolith NT.115 using LED power 30% and IR laser power 40% and 60%; Kd determined using 1:1 binding model. Hydrogen-deuterium exchange mass spectrometry (HXMS): 1 mg/ml BAX monomer or BAX with BAI1 (2 mg/ml) was diluted into D2O buffer (8.3 mM Tris, 150 mM NaCl, pDread 7.2), incubated at 0°C for 10 s, 100 s, 1000 s, quenched, frozen, and analyzed. Paramagnetic relaxation enhancement (PRE): 50 μM 15N-labeled BAX with or without 200 μM BAI1 and 10 mM hy-TEMPO, HSQC acquired with 5 s recycle delay; PRE calculated as ratio of peak intensities with/without hy-TEMPO [1].
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| Cell Assay |
Caspase 3/7 assay of WT MEFs after pro-apoptotic TNF/CHX treatment, whether BAI1 was given in conjunction with it or not at the recommended doses for 8 hours.
BAI1 was tested in cell-based assays. Wild-type MEFs, BAK KO MEFs, and BAX KO MEFs (1×10^4 cells/well in 96-well plates) were treated with TNFα (pro-apoptotic) plus cycloheximide (CHX) in the presence or absence of BAI1 at indicated doses for 8 h. Caspase-3/7 activation was measured using Caspase-Glo 3/7 chemiluminescence reagent. BAI1 inhibited apoptosis in WT MEFs (IC50 = 1.8 μM) and BAK KO MEFs but not in BAX KO MEFs. OCI-AML3 cells (2.5×10^3 cells/well in 384-well plates) were treated with BTSA1 (BAX direct activator) and BAI1 or DMSO for 2.5 h in serum-free media, then 10% FBS added; viability assayed at 6 h by CellTiter-Glo. BAI1 dose-dependently inhibited BTSA1-induced cell death. Mitochondrial depolarization assay: BAX-/- BAK-/- MEFs reconstituted with human BAX (5,000 cells/well in 384-well plate) were treated with BAI1 (5, 2.5, 1.25 μM) or DMSO for 2 h, washed, then stained with JC-1 (2 μM), oligomycin (0.02 mg/ml), digitonin (0.05 mg/ml), 2-mercaptoethanol (0.01 M) in MEB solution (150 mM mannitol, 10 mM HEPES-KOH, 50 mM KCl, 0.02 mM EGTA, 0.02 mM EDTA, 0.1% BSA, 5 mM succinate, pH 7.5), then treated with 5 μM BIM BH3 peptide; fluorescence measured at 545/590 nm; BAI1 dose-dependently inhibited BIM BH3-induced mitochondrial depolarization [1]. |
| References | |
| Additional Infomation |
1-(3,6-Dibromo-9-carbazolyl)-3-(1-piperazinyl)-2-propanol is a member of the bromobenzene class and biphenyl class.
BAI1 is a small-molecule allosteric inhibitor of BAX that binds to a novel pocket (BAI-site) at the junction of helices α3, α4, α5 and α6, distinct from the N-terminal trigger site and the canonical BH3 groove. It stabilizes the inactive conformation of BAX, preventing BH3-induced conformational changes, BH3 domain exposure, mitochondrial translocation, and oligomerization. BAI1 selectively inhibits BAX-dependent (but not BAK-dependent) cell death. It was first identified from a screen using a BAX-mediated cytochrome c release assay. Structure-activity relationship (SAR) shows that the dibromo-substituted carbazole ring and the piperazine-propanol extension are important for activity. BAI1 is a useful tool for probing BAX activation mechanisms and has therapeutic potential for diseases mediated by BAX-dependent cell death such as myocardial infarction, stroke, and neurodegeneration [1]. |
| Molecular Formula |
C19H21BR2N3O
|
|---|---|
| Molecular Weight |
540.11942
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| Exact Mass |
536.958
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| Elemental Analysis |
C, 48.85; H, 4.53; Br, 34.21; N, 8.99; O, 3.42
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| CAS # |
335165-68-9
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| Related CAS # |
335165-68-9
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| PubChem CID |
2729027
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| Appearance |
White to off-white solid powder
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| Boiling Point |
388.1ºC at 760 mmHg
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| Flash Point |
188.5ºC
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| LogP |
5.456
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
25
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| Complexity |
422
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| Defined Atom Stereocenter Count |
0
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| SMILES |
OC(CN1CCNCC1)CN2C3=C(C4=C2C=CC(Br)=C4)C=C(Br)C=C3
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| InChi Key |
KUUJEXLRLIPQQJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H21Br2N3O/c20-13-1-3-18-16(9-13)17-10-14(21)2-4-19(17)24(18)12-15(25)11-23-7-5-22-6-8-23/h1-4,9-10,15,22,25H,5-8,11-12H2
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| Chemical Name |
1-(3,6-dibromocarbazol-9-yl)-3-piperazin-1-ylpropan-2-ol
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| Synonyms |
BAI1; BAI 1; BAI-1; Brain-specific angiogenesis inhibitor 1
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO: ~18 mg/mL (~38.5 mM)
Ethanol: ~3 mg/mL (~6.4 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.87 mg/mL (4.00 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 18.7 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1.87 mg/mL (4.00 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 18.7 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8514 mL | 9.2572 mL | 18.5144 mL | |
| 5 mM | 0.3703 mL | 1.8514 mL | 3.7029 mL | |
| 10 mM | 0.1851 mL | 0.9257 mL | 1.8514 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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