CaCCinh-A01 is an inhibitor of calcium-activated chloride channel (CaCC) TMEM16A, with an IC50 value of 7.35 ± 0.86μM for TMEM16A current [1]. CaCCinh-A01, a potent blocker of Best1 channels, with no effect on intracellular Ca2+ and Ca2+-calmodulin kinase II or CFTR [2]. CaCCinh-A01 has been widely used in animal models to induce vasodilation in small resistance arteries under experimental conditions[3].
In vitro, CaCCinh-A01 treatment for 72 hours it significantly inhibited the proliferation of Te11 cells, FaDu cells, Te1 cells and HeLa cells, with IC50 values of 2.2, 5.8, 37 and 33μM respectively[4]. Treatment with 10μM CaCCinh-A01 for 72 hours significantly inhibited the viability of HT-29 cells, increased intracellular ROS levels, activated the mitochondrial apoptotic pathway, arrested the cell cycle at the S phase, and upregulated the expression of cell cycle-related proteins[5]. Treatment with 30μM CaCCinh-A01 for 24 hours enhanced the proliferation and wound healing ability of BEAS-2B cells, resulting in an increase in intracellular chloride ion content and an increase in FAK phosphorylation levels in cells[6].
In vivo, CaCCinh-A01 treatment via intraperitoneal injection (0.5mg/kg) twice daily for 14 consecutive days alleviated the intestinal damage in C57BL/6 mice caused by whole abdominal irradiation (WAI), accompanied by a decrease in the expression of Anoctamin 1 (ANO1)[7]. Male C57BL/6J mice were intraperitoneally injected with CaCCinh-A01 (17mg/kg/day) once daily for 7 consecutive days could prevent renal fibrosis caused by unilateral ureteral obstruction (UUO), and reduce the expression of fibronectin, α-SMA and collagen in the obstructed kidneys[8].
References:
[1] Shi S, Guo S, Chen Y, et al. Molecular mechanism of CaCCinh-A01 inhibiting TMEM16A channel[J]. Archives of Biochemistry and Biophysics, 2020, 695: 108650.
[2] Liu Y, Zhang H, Huang D, et al. Characterization of the effects of Cl− channel modulators on TMEM16A and bestrophin-1 Ca2+ activated Cl− channels[J]. Pflügers Archiv-European Journal of Physiology, 2015, 467(7): 1417-1430.
[3] Boedtkjer D M B, Kim S, Jensen A B, et al. New selective inhibitors of calcium‐activated chloride channels–T 16 A inh‐A 01, C a CC inh‐A 01 and MONNA–what do they inhibit?[J]. British journal of pharmacology, 2015, 172(16): 4158-4172.
[4] Bill A, Hall M L, Borawski J, et al. Small molecule-facilitated degradation of ANO1 protein[J]. Journal of Biological Chemistry, 2014, 289(16): 11029-11041.
[5] Guo S, Yan H, Wang L, et al. GlyH-101 and CaCCinh-A01 Inhibited HT-29 Proliferation by Activating the Mitochondrial Apoptosis Pathway and Arresting the Cell Cycle[J]. Anticancer Research, 2023, 43(8): 3471-3477.
[6] Wang J, Luo J, Huang W, et al. Increased intracellular Cl− concentration by activating FAK promotes airway epithelial BEAS-2B cells proliferation and wound healing[J]. Archives of Biochemistry and Biophysics, 2020, 680: 108225.
[7] Guo Y, Yuan T, Wang Y, et al. Blockade of calcium-activated chloride channel ANO1 ameliorates ionizing radiation-induced intestinal injury[J]. Journal of Advanced Research, 2025.
[8] Li X L, Liu J, Chen X S, et al. Blockade of TMEM16A protects against renal fibrosis by reducing intracellular Cl− concentration[J]. British Journal of Pharmacology, 2022, 179(12): 3043-3060.
CaCCinh-A01是一种钙激活氯通道（CaCC）TMEM16A抑制剂，对TMEM16A电流的IC50值为7.35 ± 0.86μM[1]。作为Best1通道的有效阻断剂，CaCCinh-A01不影响细胞内Ca²⁺、钙调蛋白激酶II或CFTR功能[2]。CaCCinh-A01已广泛应用于动物模型中诱导实验条件下小阻力动脉的血管舒张[3]。
在体外，CaCCinh-A01处理72小时可显著抑制Te11、FaDu、Te1和HeLa细胞增殖，IC50值分别为2.2μM、5.8μM、37μM和33μM[4]。10μM的CaCCinh-A01处理HT-29细胞72小时能抑制细胞活力，增加细胞内ROS水平，激活线粒体凋亡通路，使细胞周期阻滞于S期，并上调周期相关蛋白表达[5]。30μM的CaCCinh-A01处理BEAS-2B细胞24小时可yiz细胞增殖和伤口愈合能力，增加细胞内氯离子含量并提高FAK磷酸化水平[6]。
在体内，C57BL/6小鼠每日两次腹腔注射CaCCinh-A01（0.5mg/kg；持续14天）可减轻全腹部照射（WAI）引起的肠道损伤，并伴随Anoctamin 1（ANO1）表达下降[7]。雄性C57BL/6J小鼠每日腹腔注射17mg/kg CaCCinh-A01（持续7天）能预防单侧输尿管梗阻（UUO）引起的肾纤维化，并降低梗阻肾脏中纤连蛋白、α-SMA和胶原的表达[8]。