BAM 7 is a Bax activator (EC50 = 3.3 μM). BCL-2 family proteins are key regulators of the apoptotic pathway. In order to preserve cell survival, antiapoptotic members bind proapoptotic members' BCL-2 homology 3 (BH3) death domains, such as BAX. To successfully reactivate apoptosis in cancer, the antiapoptotic BH3-binding groove has been targeted.
Physicochemical Properties
| Molecular Formula | C21H19N5O2S |
| Molecular Weight | 405.476 |
| Exact Mass | 405.125 |
| Elemental Analysis | C, 62.21; H, 4.72; N, 17.27; O, 7.89; S, 7.91 |
| CAS # | 331244-89-4 |
| Related CAS # | 331244-89-4 |
| PubChem CID | 3101542 |
| Appearance | Pink to red solid powder |
| Density | 1.3±0.1 g/cm3 |
| Boiling Point | 567.3±52.0 °C at 760 mmHg |
| Flash Point | 296.9±30.7 °C |
| Vapour Pressure | 0.0±1.6 mmHg at 25°C |
| Index of Refraction | 1.685 |
| LogP | 3.18 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 29 |
| Complexity | 649 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | CCOC1=CC=CC=C1N/N=C2C(C)=NN(C3=NC(C4=CC=CC=C4)=CS3)C\2=O |
| InChi Key | WRLVHADVOGFZOZ-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C21H19N5O2S/c1-3-28-18-12-8-7-11-16(18)23-24-19-14(2)25-26(20(19)27)21-22-17(13-29-21)15-9-5-4-6-10-15/h4-13,25H,3H2,1-2H3 |
| Chemical Name | 4-[(2-ethoxyphenyl)diazenyl]-5-methyl-2-(4-phenyl-1,3-thiazol-2-yl)-1H-pyrazol-3-one |
| Synonyms | BAM 7; BAM7; BAM-7 |
| 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 (IC50 = 3.3 μM) |
| ln Vitro | BAM7 directly binds the previously uncharacterized BH3-binding groove at the N-terminal face of BAX. At the N-terminal face of BAX, the BH3-binding groove is specifically recognized by BAM7. In fact, BAM7 directly interacts with BAX at the surface where the BIM BH3 helix activates BAX. Functional activation of BAX is the result of BAM7. In a dose- and time-responsive manner, BAM7 causes BAX to change from monomer to oligomer, with the kinetics approaching saturation at a dose ratio of 1:8 for BAX:BAM7. In vitro BAX oligomerization, BAX-mediated pore formation, and BAX-dependent cell death are all sparked by BAM7. BAM7 selectively induces BAX-mediated apoptosis by triggering the hallmark features of intracellular BAX activation. The biochemical and morphological characteristics of BAX-mediated apoptosis are only induced by BAM7 in the cell line that contains BAX. [1] |
| Enzyme Assay | In order to create direct binding curves, FITC-BIM SAHB (50 nM) is first incubated with serial dilutions of full-length BAX, BCL-XLΔC, MCL-1ΔNΔC, BFL-1/A1ΔC or BAKΔC, or BAKC, with fluorescence polarization being measured after 20 minutes on a SpectraMax M5 microplate reader. For competition assays, small molecules or acetylated BIM SAHB (Ac-BIM SAHB) are mixed with FITC-BIM SAHB (50 nM), and then recombinant protein is added at an ~EC75 concentration (BAX, BAKΔC: 500 nM; BCL-XLΔC, MCL-1ΔNΔC, BFL-1/A1ΔC: 200 nM) determined by the direct binding assay. IC50 values are determined by nonlinear regression analysis of competitive binding curves using Prism software after 20 minutes of fluorescence polarization measurement. |
| Cell Assay | MEFs (2.5 × 103 cells per well) are seeded in 96-well opaque plates for 18–24 h, after which they are incubated at 37 °C in a final volume of 100 L with serial dilutions of BAM7, ANA-BAM16, or vehicle (0.15% (v/v) DMSO). According to the manufacturer's instructions, CellTiter-Glo reagent is added to assay cell viability at 24 hours, and luminescence is measured using a SpectraMax M5 microplate reader. At least three replicates of the viability assays are carried out, and the data are then normalized to the vehicle-treated control wells. |
| References |
[1]. Direct and selective small-molecule activation of proapoptotic BAX. Nat Chem Biol. 2012 Jul;8(7):639-45. |
Solubility Data
| Solubility (In Vitro) | DMSO: 2~5 mg/mL (4.9~12.3 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 0.5 mg/mL (1.23 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 5.0 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 | 2.4662 mL | 12.3311 mL | 24.6621 mL | |
| 5 mM | 0.4932 mL | 2.4662 mL | 4.9324 mL | |
| 10 mM | 0.2466 mL | 1.2331 mL | 2.4662 mL |