BAI1 is a novel, potent and direct allosteric inhibitor of BAX with a dissociation constant (Kd) of 15.0 ± 4 μM. In both healthy and unhealthy contexts, BAX plays a crucial role in the mitochondrial cell death pathway's response to a variety of stimuli. Cytosolic BAX is activated and translocated in a conformational manner upon binding by BH3-only proteins, which causes the permeabilization of the mitochondrial outer membrane. Despite the obvious therapeutic potential of preventing BAX-mediated cell death in a variety of diseases, efforts to rationally target BAX and develop inhibitors have proven difficult. BAX activation conformational events that prevent mitochondrial translocation and oligomerization are inhibited by BAIs. Our findings point to a novel paradigm for the efficient and targeted pharmacological targeting of BAX to support the rational design of BAX-mediated cell death inhibitors.

Physicochemical Properties

Molecular Formula	C19H21BR2N3O
Molecular Weight	540.11942
Exact Mass	536.958
Elemental Analysis	C, 48.85; H, 4.53; Br, 34.21; N, 8.99; O, 3.42
CAS #	335165-68-9
Related CAS #	335165-68-9
PubChem CID	2729027
Appearance	White to off-white solid powder
Boiling Point	388.1ºC at 760 mmHg
Flash Point	188.5ºC
LogP	5.456
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	4
Heavy Atom Count	25
Complexity	422
Defined Atom Stereocenter Count	0
SMILES	OC(CN1CCNCC1)CN2C3=C(C4=C2C=CC(Br)=C4)C=C(Br)C=C3
InChi Key	KUUJEXLRLIPQQJ-UHFFFAOYSA-N
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
Chemical Name	1-(3,6-dibromocarbazol-9-yl)-3-piperazin-1-ylpropan-2-ol
Synonyms	BAI1; BAI 1; BAI-1; Brain-specific angiogenesis inhibitor 1
HS Tariff Code	2934.99.9001
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)

Biological Activity

Targets	Bax (Kd = 15.0 μM)
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].
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.
References	[1]. Small-molecule allosteric inhibitors of BAX. Nat Chem Biol. 2019 Apr;15(4):322-330.
Additional Infomation	1-(3,6-dibromo-9-carbazolyl)-3-(1-piperazinyl)-2-propanol is a member of bromobenzenes and a member of biphenyls.

Solubility Data

Solubility (In Vitro)

DMSO: ~18 mg/mL (~38.5 mM) Ethanol: ~3 mg/mL (~6.4 mM)

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.