Background
BAM15 is a novel mitochondrial protonophore uncoupler (2-fluorophenyl){6-[(2-fluorophenyl)amino](1,2,5-oxadiazolo[3,4-e]pyrazin-5-yl)}amine, that does not depolarize the plasma membrane[1].
BAM15 has an average EC50 value of 1.4 μM and is capable of maintaining high levels of mitochondrial respiration over a wide concentration range of 3 to 100 μM in OCR[2]. In fully differentiated C2C12 mouse myotubes, BAM15 (100 μM;16h) did not alter cell viability or number,but BAM15 improves cellular respiratory kinetics by sustained mitochondrial uncoupling[3]. Wild-type animals treated with both 50 μM BAM15 showed reduced mechanosensory neuronal defects during aging, which correlates with the maintenance of touch responses and short-term memory during aging in Caenorhabditis elegans[4]. BAM15(1mM) depolarized mitochondrial membrane potential, induced mitochondrial fission, increased mitochondrial ROS production, and increased mitochondrial oxygen consumption rate in A10 cells[5].
BAM15(1 mg/kg; i.p.; 3 hours before injection of LPS) attenuated serum cytokines, organ damage (liver enzymes and serum creatinine) and tissue cytokines (liver and kidney), and reduced inflammatory mononuclear/macrophages in the liver, in part by enhancing phosphorylated αAMPK in LPS-administered mice[6]. BAM15 was seen to reduce mortality after injection at 0, 6 and 12 hours after cecal ligation and puncture (CLP), and reduced kidney damage and spleen apoptosis even after 12 hours when the mice were sick[7].
BAM15是一种新型线粒体质子解耦解偶联剂(2-氟苯基){6-[(2-氟苯基)氨基](1,2,5-恶二唑啉[3,4-e]吡嗪-5-基)}胺,不去极化质膜[1]。
BAM15在刺激耗氧率(OCR)方面展现出了强大的能力,BAM15平均EC50值为1.4 μM,并且能够在3 ~ 100 μM的大浓度范围内维持高水平的线粒体呼吸[2]。在完全分化的C2C12小鼠肌管中,BAM15 (100 μM;16h)不改变细胞活力或数量,但会通过持续的线粒体解耦改善细胞呼吸动力学[3]。用50 μM BAM15处理的野生型秀丽隐杆线虫在衰老过程中表现出机械感觉神经元缺陷的减少,这与衰老过程中触摸反应和短期记忆的维持有关[4]。BAM15能有效激活A10细胞中的AMPK,且BAM15的作用强于CCCP、氯硝柳胺、AMPK阳性激活剂二甲双胍和AICAR[5]。
BAM15在一定程度上通过增强磷酸化的αAMPK减弱了肝脏中的血清细胞因子,器官损伤(肝酶和血清肌酐)和组织细胞因子(肝脏和肾脏),并减少了肝脏中的炎性单核/巨噬细胞[6]。BAM15在盲肠结扎和穿刺(CLP)后0,6和12小时注射后降低死亡率,即使在小鼠生病12小时后也能减少肾脏损伤和脾凋亡[7]。
参考文献:
[1]. Kenwood BM, Weaver JL, et,al. Identification of a novel mitochondrial uncoupler that does not depolarize the plasma membrane. Mol Metab. 2013 Nov 28;3(2):114-23. doi: 10.1016/j.molmet.2013.11.005. PMID: 24634817; PMCID: PMC3953706.
[2]. Alexopoulos SJ, Chen SY, et,al. Mitochondrial uncoupler BAM15 reverses diet-induced obesity and insulin resistance in mice. Nat Commun. 2020 May 14;11(1):2397. doi: 10.1038/s41467-020-16298-2. PMID: 32409697; PMCID: PMC7224297.
[3]. Axelrod CL, King WT, et,al. BAM15-mediated mitochondrial uncoupling protects against obesity and improves glycemic control. EMBO Mol Med. 2020 Jul 7;12(7):e12088. doi: 10.15252/emmm.202012088. Epub 2020 Jun 10. PMID: 32519812; PMCID: PMC7338798.
[4]. Cho I, Song HO, et,al. BAM15 Relieves Neurodegeneration in Aged Caenorhabditis elegans and Extends Lifespan. Metabolites. 2022 Nov 17;12(11):1129. doi: 10.3390/metabo12111129. PMID: 36422268; PMCID: PMC9698188.
[5]. Tai Y, Li L, et,al. Mitochondrial uncoupler BAM15 inhibits artery constriction and potently activates AMPK in vascular smooth muscle cells. Acta Pharm Sin B. 2018 Oct;8(6):909-918. doi: 10.1016/j.apsb.2018.07.010. Epub 2018 Jul 26. PMID: 30505660; PMCID: PMC6251816.
[6]. Dang CP, Issara-Amphorn J, et,al. BAM15, a Mitochondrial Uncoupling Agent, Attenuates Inflammation in the LPS Injection Mouse Model: An Adjunctive Anti-Inflammation on Macrophages and Hepatocytes. J Innate Immun. 2021;13(6):359-375. doi: 10.1159/000516348. Epub 2021 Jun 1. PMID: 34062536; PMCID: PMC8613553.
[7]. Tsuji N, Tsuji T, et,al. BAM15 treats mouse sepsis and kidney injury, linking mortality, mitochondrial DNA, tubule damage, and neutrophils. J Clin Invest. 2023 Feb 9:e152401. doi: 10.1172/JCI152401. Epub ahead of print. PMID: 36757801.
Protocol
| Cell experiment [1]: |
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Cell lines
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Normal murine liver (NMuLi) cells
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Preparation Method
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NMuLi cells were seeded in a 96-well seahorse plates and were allowed to adhere overnight. Prior to the assay, media was changed to unbuffered DMEM containing pyruvate and glutamine and the cells were equilibrated for 1 h at 37℃ without CO2. BAM15 were injected during the assay and OCR(Oxygen consumption rate) was measured using 2 min measurement periods.
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Reaction Conditions
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0.1-100 μM
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Applications
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BAM15 has an average EC50 value of 1.4 μM and is capable of maintaining high levels of mitochondrial respiration over a wide concentration range of 3 to 100 μM in OCR.
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| Animal experiment [2]: |
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Animal models
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Male 6-week-old mice
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Preparation Method
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BAM15 at 1 mg/kg in 10% of DMSO or 10% DMSO alone was intraperitoneally administered at 3 h prior to the intraperitoneal injection of LPS (4 mg/kg) in PBS or PBS alone.
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Dosage form
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1 mg/kg; i.p.; 3 hours before injection of LPS
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Applications
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BAM15 attenuated serum cytokines, organ damage (liver enzymes and serum creatinine) and tissue cytokines (liver and kidney), and reduced inflammatory mononuclear/macrophages in the liver, in part by enhancing phosphorylated αAMPK.
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参考文献:
[1]. Alexopoulos SJ, Chen SY, et,al. Mitochondrial uncoupler BAM15 reverses diet-induced obesity and insulin resistance in mice. Nat Commun. 2020 May 14;11(1):2397. doi: 10.1038/s41467-020-16298-2. PMID: 32409697; PMCID: PMC7224297.
[2]. Dang CP, Issara-Amphorn J, et,al. BAM15, a Mitochondrial Uncoupling Agent, Attenuates Inflammation in the LPS Injection Mouse Model: An Adjunctive Anti-Inflammation on Macrophages and Hepatocytes. J Innate Immun. 2021;13(6):359-375. doi: 10.1159/000516348. Epub 2021 Jun 1. PMID: 34062536; PMCID: PMC8613553.
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