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| 5mg |
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| 25mg |
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| 100mg |
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Purity: ≥98%
BI-6C9 is a novel and potent inhibitor of BH3 interacting domain (Bid) with Kd of 20 µM, it acts by preventing mitochondrial outer membrane potential (MOMP) and mitochondrial fission, and protecting the cells from cell death. The Bid-inhibitor BI-6c9 prevented MOMP and mitochondrial fission, and protected the cells from cell death. Oxidative stress induced by glutamate causes mitochondrial translocation of Bid thereby inducing mitochondrial fission and associated mitochondrial cell death pathways. Inhibiting regulators of pathological mitochondrial fragmentation is proposed as an efficient strategy of neuroprotection.
| Targets |
BI-6c9 is a highly specific inhibitor of the pro-apoptotic Bcl-2 family protein Bid.[1]
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| ln Vitro |
About 4.5 hours after glutamate (3 mM and 5 mM) treatment, HT-22 cells displayed decreased cell viability. The loss of cell viability following glutamate toxicity was greatly reduced by BI-6c9 (10 μM; 18 hours) [1].
In HT-22 hippocampal neurons exposed to glutamate (5 mM), treatment with BI-6c9 (10 µM) prevented glutamate-induced mitochondrial fission, preserving the tubular network morphology (Categories I & II) and preventing the shift to fragmented mitochondria (Categories III & IV).[1] BI-6c9 (10 µM) prevented the breakdown of mitochondrial membrane potential (ΔΨm) induced by glutamate (5 mM) in HT-22 cells, as assessed by JC-1 fluorescence. It restored the red/green fluorescence ratio to near control levels, comparable to the protection seen against the depolarizing agent CCCP.[1] BI-6c9 (10 µM) attenuated glutamate-induced cell death in HT-22 neurons. In MTT assays, it restored cell viability from ~20% (with glutamate alone) to nearly baseline levels. Real-time impedance monitoring (xCELLigence System) confirmed that BI-6c9 prevented the delayed decrease in Normalized Cell Index (NCI) caused by glutamate (3 or 5 mM), indicating protection against cell death.[1] |
| Cell Assay |
Cell viability assay [1]
Cell Types: HT-22 cells Tested Concentrations: 10 μM Incubation Duration: 18 hrs (hours) Experimental Results: Attenuated glutamate toxicity Acid-induced cell death. Cell Viability (MTT Assay): HT-22 cells were seeded in 96-well plates. After treatment with glutamate and/or BI-6c9 for 18 hours, MTT was added to a final concentration of 0.25 mg/ml and incubated for 2 hours. The reaction was stopped by removing the medium, freezing the plate at -80°C for at least 1 hour, and adding dimethylsulfoxide to dissolve the formazan crystals. Absorbance was measured at 590 nm with a reference at 630 nm.[1] Mitochondrial Morphology Analysis: HT-22 cells were transfected with a mitochondria-targeted GFP (mGFP) plasmid. 24 hours post-transfection, cells were seeded onto coated slides and treated. After 18 hours, cells were fixed with 4% PFA, nuclei were counterstained with DAPI, and images were acquired by fluorescence or confocal microscopy. Mitochondria were categorized into four morphological states (elongated/tubular to fragmented) by a blinded investigator, and mitochondrial length was quantified using ImageJ software.[1] Mitochondrial Membrane Potential (JC-1 Assay): After treatment (e.g., 12 hours with glutamate), HT-22 cells were stained with JC-1 dye (2 µM final concentration) according to the protocol. Cells were collected, washed, and resuspended in PBS. JC-1 fluorescence was analyzed by flow cytometry or epifluorescence microscopy. Green fluorescence (monomeric form, ~530 nm emission) indicates dye uptake, while red fluorescence (J-aggregates, ~613 nm emission) indicates intact ΔΨm. The positive control for depolarization was treatment with CCCP.[1] Real-time Cell Impedance Monitoring (xCELLigence System): HT-22 cells were seeded at 4500 cells/well in specialized 96-well E-plates containing integrated microelectrodes. After 24 hours, cells were treated with vehicle, BI-6c9 (10 µM), glutamate (3 or 5 mM), or their combination. The electrical impedance across the electrodes, reported as the Normalized Cell Index (NCI), was continuously monitored in real-time without removing the plate from the incubator. A decrease in NCI correlates with cell death/detachment.[1] |
| References | |
| Additional Infomation |
BI-6c9 is known as a "highly specific Bid inhibitor". This study proposes that inhibiting regulators of pathological mitochondrial cleavage such as Bid through BI-6c9 is an effective neuroprotective strategy. Its mechanism involves BI-6c9 preventing the activation/translocation of Bid to mitochondria, thereby inhibiting downstream events: mitochondrial division, loss of mitochondrial membrane potential, increased mitochondrial outer membrane permeability, and the release of apoptosis factors such as AIF, ultimately preventing caspase-independent neuronal cell death. [1]
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| Molecular Formula |
C23H25N3O4S2
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|---|---|
| Molecular Weight |
471.5923
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| Exact Mass |
471.129
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| Elemental Analysis |
C, 58.58; H, 5.34; N, 8.91; O, 13.57; S, 13.60
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| CAS # |
791835-21-7
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| PubChem CID |
11547341
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| Appearance |
White to off-white solid powder
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| Density |
1.368g/cm3
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| Index of Refraction |
1.671
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| LogP |
6.251
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
32
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| Complexity |
661
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(CCCNS(C1C=CC(OC)=CC=1)(=O)=O)NC1C=CC(SC2C=CC(N)=CC=2)=CC=1
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| InChi Key |
LCFUJBSKPDPGKO-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C23H25N3O4S2/c1-30-19-8-14-22(15-9-19)32(28,29)25-16-2-3-23(27)26-18-6-12-21(13-7-18)31-20-10-4-17(24)5-11-20/h4-15,25H,2-3,16,24H2,1H3,(H,26,27)
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| Chemical Name |
N-[4-[(4-aminophenyl)thio]phenyl]-4-[[(4-methoxyphenyl)sulfonyl]amino]-butanamide
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| Synonyms |
BI-6C9; BI 6C9; BI6C9
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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 : ~250 mg/mL (~530.12 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.41 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 20.8 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: ≥ 2.08 mg/mL (4.41 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (4.41 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1205 mL | 10.6024 mL | 21.2049 mL | |
| 5 mM | 0.4241 mL | 2.1205 mL | 4.2410 mL | |
| 10 mM | 0.2120 mL | 1.0602 mL | 2.1205 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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