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BAM15

  Cat. No.:  DC32783   Featured
Chemical Structure
210302-17-3
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More than 5000 active chemicals with high quality for research!
Field of application
BAM15 is a potent and selective mitochondrial uncoupler or protonophore. Chemical mitochondrial uncouplers are lipophilic weak acids that transport protons into the mitochondrial matrix via a pathway that is independent of ATP synthase, thereby uncoupling nutrient oxidation from ATP production. These uncouplers have potential for the treatment of diseases such as obesity, Parkinson’s disease, and aging.
Cas No.: 210302-17-3
Chemical Name: N5,N6-bis(2-fluorophenyl)-[1,2,5]oxadiazolo[3,4-b]pyrazine-5,6-diamine
Synonyms: BAM15; BAM-15; BAM 15.
SMILES: FC1=CC=CC=C1NC2=NC3=NON=C3N=C2NC4=C(C=CC=C4)F
Formula: C16H10F2N6O
M.Wt: 340.29
Purity: >98%
Sotrage: 2 years -20°C Powder, 2 weeks 4°C in DMSO, 6 months -80°C in DMSO
Description: BAM 15 is a novel mitochondrial protonophore uncoupler.
In Vivo: Compare to vehicle-treated mice, animals that receive BAM 15 are protected from kidney injury as indicated by lower plasma creatinine levels at 24 and 48 h post-ischemia, reduced tubular necrosis, less depletion of brush border villi, less obstruction of proximal tubules, and less immune cell infiltration[1].
In Vitro: BAM 15 is able to increase O2 consumption across a broad dosing range without increasing ROS. BAM 15 and FCCP are structurally unrelated and it is observed that low doses of BAM 15 from 100 nM to 1 μM increase cellular O2 consumption rate (OCR) to a similar degree as FCCP, but higher concentrations from 1 μM to 50 μM reveal that BAM 15 is able to maintain uncoupled respiration at a high rate in a range of cell lines. BAM 15 is fully capable of increasing mitochondrial respiration in the presence of oligomycin and does so across a broader concentration range than FCCP in both myoblasts and hepatocytes. BAM 15 induces mitochondrial swelling, demonstrating that BAM 15 is a protonophore. BAM15-treated cells are more viable than FCCP-treated cells when administered across a broad dosing range up to 50 μM[1].
Kinase Assay: Electron flow assays are performed. Briefly, 5 μg of mitochondrial protein in MAS is loaded into a 24-well tissue culture plate and centrifuged at 2000×g for 15 min at 4°C. Prior to the assay, mitochondria are incubated at 37°C for 10 mins in MAS containing 10 mM pyruvate, 2 mM malate, and 5 μM BAM 15 or FCCP. Rotenone (2 μM), succinate (10 mM), antimycin A (4 μM), and N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD, 100 μM) plus ascorbate (10 mM) are added sequentially as indicated in the figure. N=3 wells/plate of a representative of 3 plates[1].
Cell Assay: L6 cells are incubated with the fluorescent indicator of mitochondrial membrane potential tetramethylrhodamine (TMRM, 125 nM) or DMSO (1%) control for 30 min. The cells are then centrifuged for 5 min at 700×g and resuspended in unbuffered DMEM at a concentration of 1×105 cells/mL. The cells are then treated with BAM 15 or DMSO (0.1%) for 10 min prior to flow cytometric analysis[1].
Animal Administration: Male mice (8-week old, C57BL/6) are anesthetized with a mixture (i.p.) of ketamine (120 mg/kg), xylazine (12 mg/kg), and atropine (0.324 mg/kg) and are subjected to bilateral ischemic reperfusion injury (26 min ischemia, then 24 h or 48 h reperfusion). During the surgery, mouse core temperature is maintained at 34 to 36°C with a heating pad; during the recovery and reperfusion period, mice are housed in a warming incubator with ambient temperature at 30 to 32 °C. Control, sham-operated mice undergo a similar procedure, but the renal pedicles are not clamped. Mice are i.p. injected with BAM 15 at 1 or 5 mg/kg, 1 h before kidney IR. Vehicle mice are also injected with the same solution BAM 15 is prepared with (3% DMSO in 50% PEG400)[1].
References: [1]. Kenwood BM, et al. Identification of a novel mitochondrial uncoupler that does not depolarize the plasma membrane. Mol Metab. 2013 Nov 28;3(2):114-23.
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