Species Comparison and Pharmacological Characterization of Human, Monkey, Rat, and Mouse TRPA1 Channels
Species Comparison and Pharmacological Characterization of Human, Monkey, Rat, and Mouse TRPA1 Channels
复制标题
DOI：
10.1124/jpet.111.189902
复制
发表时间：
2012-05-01
影响因子：
3.5
通讯作者：
Chen, Jun
中科院分区：
文献类型：
--
作者：
Bianchi, Bruce R.;Zhang, Xu-Feng;Chen, Jun
瞬时受体电位锚蛋白-1（TRPA 1）通道已成为开发镇痛和抗炎药物的有吸引力的靶点。然而，靶向TRPA 1的药物发现工作受到人类和啮齿动物物种之间差异的阻碍。许多化合物已被鉴定对人TRPA 1（hTRPA 1）具有拮抗剂活性，但当对大鼠TRPA 1（rTRPA 1）和小鼠TRPA 1（mTRPA 1）进行测试时，它们显示出作为拮抗剂的效力降低、无作用或激动剂活性。这些化合物被排除在进一步的药物开发之外，因为它们不能在使用常规大鼠/小鼠模型的临床前研究中进行测试。为了扩大我们对物种特异性差异的理解，我们克隆了恒河猴TRPA 1（rhTRPA 1）并对其功能进行了表征，并将其药理学特征与hTRPA 1，rTRPA 1和mTRPA 1通道进行了比较。使用瞬时转染的人胚肾293-F细胞，在荧光Ca 2+内流测定中测定了TRPA 1配体（反应性和非反应性）的不同组的功能活性。4-甲基-N-[2，2，2-三氯-1-（4-硝基-苯硫基）-乙基]-苯甲酰胺、薄荷醇和咖啡因在TRPA 1处显示出种属特异性差异药理学。发现rhTRPA 1通道的药理学特征与hTRPA 1通道相似。相比之下，rTRPA 1和mTRPA 1通道彼此非常相似，但与hTRPA 1或rhTRPA 1通道截然不同。我们的研究结果表明，TRPA 1功能在灵长类动物和啮齿类动物之间存在差异，并表明恒河猴可以在临床前研究中作为人类的替代物种。
The transient receptor potential ankyrin-1 (TRPA1) channel has emerged as an attractive target for development of analgesic and anti-inflammatory drugs. However, drug discovery efforts targeting TRPA1 have been hampered by differences between human and rodent species. Many compounds have been identified to have antagonist activity at human TRPA1 (hTRPA1), but when tested at rat TRPA1 (rTRPA1) and mouse TRPA1 (mTRPA1), they show reduced potency as antagonists, no effect, or agonist activity. These compounds are excluded from further drug development because they cannot be tested in preclinical studies using conventional rat/mouse models. To broaden our understanding of species-specific differences, we cloned and functionally characterized rhesus monkey TRPA1 (rhTRPA1) and compared its pharmacological profile to hTRPA1, rTRPA1, and mTRPA1 channels. The functional activities of a diverse group of TRPA1 ligands (both reactive and nonreactive) were determined in a fluorescent Ca2+ influx assay, using transiently transfected human embryonic kidney 293-F cells. 4-Methyl-N-[2,2,2-trichloro-1-(4-nitro-phenylsulfanyl)-ethyl]-benzamide, menthol, and caffeine displayed species-specific differential pharmacology at TRPA1. The pharmacological profile of the rhTRPA1 channel was found to be similar to the hTRPA1 channel. In contrast, the rTRPA1 and mTRPA1 channels closely resembled each other but were pharmacologically distinct from either hTRPA1 or rhTRPA1 channels. Our findings reveal that TRPA1 function differs between primate and rodent species and suggest that rhesus monkey could serve as a surrogate species for humans in preclinical studies.