Effect of non-steroidal anti-inflammatory drugs and new fenamate analogues on TRPC4 and TRPC5 channels
Effect of non-steroidal anti-inflammatory drugs and new fenamate analogues on TRPC4 and TRPC5 channels
复制标题
DOI：
10.1016/j.bcp.2012.01.014
复制
发表时间：
2012-04-01
影响因子：
5.8
通讯作者：
Xu, Shang-Zhong
中科院分区：
文献类型：
--
作者：
Jiang, Hongni;Zeng, Bo;Xu, Shang-Zhong
非甾体类抗炎药（NSAIDs）是目前广泛应用的抗炎治疗药物，其中芬那酯类似物在调节细胞内钙瞬变和离子通道中发挥重要作用。然而，NSAID对TRPC 4和TRPC 5的影响仍然未知。为了解芬那酯类似物对TRPC通道的构效关系，我们合成了一系列芬那酯类似物，并研究了它们对TRPC 4和TRPC 5通道的作用。膜片钳和钙离子成像分别记录全细胞电流和钙离子运动，氟芬那酸（FFA）、甲芬那酸（MFA）、尼氟灭酸（NFA）和双氯芬酸钠（DFS）对TRPC 4和TRPC 5通道均表现出浓度依赖性的抑制作用。效价为FFA > MFA > NFA > DFS。通过用-F、-CH 3、-OCH 3、-OCH 2CH 3、-COOH和-NO2取代FFA中的三氟甲基来修饰2-苯氨基环，导致其通道阻断活性的变化。然而，2-（2 '-甲氧基-5'-甲基苯基）氨基苯甲酸刺激TRPC 4和TRPC 5通道。选择性COX 1 -3抑制剂（阿司匹林、塞来昔布、对乙酰氨基酚和吲哚美辛）对通道没有影响。FFA（100 μ M）和MFA（100 μ M）长时间灌注（>5 min）后，TRPC 4和TRPC 5电流增强，这似乎部分由线粒体Ca 2+释放介导。2-苯氨基环的取代模式和构象可以改变它们的阻断活性，这对于理解芬那酯药理学和靶向TRPC通道的新药开发具有重要意义。(c)2012 Elsevier Inc. All rights reserved.
Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used anti-inflammatory therapeutic agents, among which the fenamate analogues play important roles in regulating intracellular Ca2+ transient and ion channels. However, the effect of NSAIDs on TRPC4 and TRPC5 is still unknown. To understand the structure-activity of fenamate analogues on TRPC channels, we have synthesized a series of fenamate analogues and investigated their effects on TRPC4 and TRPC5 channels.Human TRPC4 and TRPC5 cDNAs in tetracycline-regulated vectors were transfected into HEK293 T-REx cells. The whole cell current and Ca2+ movement were recorded by patch clamp and calcium imaging, respectively.Flufenamic acid (FFA), mefenamic acid (MFA), niflumic acid (NFA) and diclofenac sodium (DFS) showed inhibition on TRPC4 and TRPC5 channels in a concentration-dependent manner. The potency was FFA > MFA > NFA > DFS. Modification of 2-phenylamino ring by substitution of the trifluoromethyl group in FFA with -F, -CH3, -OCH3, -OCH2CH3, -COOH, and -NO2 led to the changes in their channel blocking activity. However, 2-(2'-methoxy-5'-methylphenyl)aminobenzoic acid stimulated TRPC4 and TRPC5 channels. Selective COX1-3 inhibitors (aspirin, celecoxib, acetaminophen, and indomethacin) had no effect on the channels. Longer perfusion (>5 min) with FFA (100 mu M) and MFA (100 mu M) caused a potentiation of TRPC4 and TRPC5 currents after their initial blocking effects that appeared to be partially mediated by the mitochondrial Ca2+ release.Our results suggest that fenamate analogues are direct modulators of TRPC4 and TRPC5 channels. The substitution pattern and conformation of the 2-phenylamino ring could alter their blocking activity, which is important for understanding fenamate pharmacology and new drug development targeting the TRPC channels. (c) 2012 Elsevier Inc. All rights reserved.