5-(N,N-Hexamethylene)-amiloride is a potent inhibitor of the Na+/H+ exchanger and is toxic to tumor cells [1]. 5-(N,N-Hexamethylene)-amiloride can inhibit DNA synthesis in regenerating liver and induce cell apoptosis, while activating the activity of Jun N-terminal kinase (JNK)[2]. 5-(N,N-Hexamethylene)-amiloride has been widely used to inhibit the growth of Cryptococcus strains and as a starting point for antifungal drug development to develop new analogues with high antifungal activity[3]. 5-(N,N-Hexamethylene)-amiloride has fluorescent properties and can regulate intracellular acidity, which can be used as a self-biomarker of pH changes in cellular studies[4].
In vitro, 5-(N,N-hexamethylene)-amiloride treatment for 48 hours inhibited the replication of mouse hepatitis virus (MHV) and human coronavirus 229E (HCoV-229E) in L929 cells, with EC50 values of 3.91μM and 1.34μM, respectively[5]. Treatment of RPMI-8226 cells with 20μM 5-(N,N-hexamethylene)-amiloride for 48 hours led to cell proliferation inhibition, and caused intracellular acidification[6]. 80µM of 5-(N,N-hexamethylene)-amiloride was applied for 24 hours to induce programmed necrosis in the cultured MCF7 cells through lysosomal and reactive oxygen species (ROS)-dependent mechanisms[7].
In vivo, 5-(N,N-hexamethylene)-amiloride treatment via intraperitoneal injection at a dose of 10mg/kg/day for 14 days suppressed tumor growth in the RPMI-8226 cell xenograft mouse models without affecting body weight[5].
References:
[1] Luo J, Tannock I F. Inhibition of the regulation of intracellular pH: potential of 5-(N, N-hexamethylene) amiloride in tumour-selective therapy[J]. British journal of cancer, 1994, 70(4): 617-624.
[2] Suzuki T, Tsukamoto I. Apoptosis induced by 5-(N, N-hexamethylene)-amiloride in regenerating liver after partial hepatectomy[J]. European journal of pharmacology, 2004, 503(1-3): 1-7.
[3] Vu K, Buckley B J, Bujaroski R S, et al. Antifungal activity of 6-substituted amiloride and hexamethylene amiloride (HMA) analogs[J]. Frontiers in Cellular and Infection Microbiology, 2023, 13: 1101568.
[4] Giansanti V, Santamaria G, Torriglia A, et al. Fluorescence properties of the Na+/H+ exchanger inhibitor HMA (5-(N, N-hexamethylene) amiloride) are modulated by intracellular pH[J]. European Journal of Histochemistry: EJH, 2012, 56(1): e3.
[5] Wilson L, Gage P, Ewart G. Hexamethylene amiloride blocks E protein ion channels and inhibits coronavirus replication[J]. Virology, 2006, 353(2): 294-306.
[6] Yang N, Dong Z, Xiao W, et al. Hexamethylene amiloride induces lysosome-mediated cell death in multiple myeloma through transcription factor E3[J]. Cell Death Discovery, 2024, 10(1): 505.
[7] Rowson-Hodel A R, Berg A L, Wald J H, et al. Hexamethylene amiloride engages a novel reactive oxygen species-and lysosome-dependent programmed necrotic mechanism to selectively target breast cancer cells[J]. Cancer letters, 2016, 375(1): 62-72.
5-(N,N-Hexamethylene)-amiloride是一种强效的Na+/H+交换体抑制剂，对肿瘤细胞具有毒性[1]。5-(N,N-Hexamethylene)-amiloride可抑制再生肝中的DNA合成并诱导细胞凋亡，同时激活Jun N末端激酶（JNK）的活性[2]。5-(N,N-Hexamethylene)-amiloride已被广泛用于抑制隐球菌菌株的生长，并作为抗真菌药物开发的起点，以开发具有高抗真菌活性的新型类似物[3]。5-(N,N-Hexamethylene)-amiloride具有荧光特性，可调节细胞内酸度，在细胞研究中可用作pH变化的自身生物标志物[4]。
在体外，5-(N,N-Hexamethylene)-amiloride处理48小时抑制了小鼠肝炎病毒（MHV）和人冠状病毒229E（HCoV-229E）在L929细胞中的复制，EC50值分别为3.91µM和1.34µM[5]。使用20µM的5-(N,N-Hexamethylene)-amiloride处理RPMI-8226细胞48小时，导致细胞增殖受抑制，并引起细胞内酸化[6]。使用80µM的5-(N,N-Hexamethylene)-amiloride处理24小时，通过溶酶体和活性氧（ROS）依赖的机制诱导培养的MCF7细胞发生程序性坏死[7]。
在体内，每日腹腔注射10mg/kg的5-(N,N-Hexamethylene)-amiloride，持续14天，抑制了RPMI-8226细胞异种移植小鼠模型中的肿瘤生长，且未影响体重[5]。