Comparative pharmacology of human β-adrenergic receptor subtypes -: characterization of stably transfected receptors in CHO cells
Comparative pharmacology of human β-adrenergic receptor subtypes -: characterization of stably transfected receptors in CHO cells
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DOI：
10.1007/s00210-003-0860-y
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发表时间：
2004-02-01
影响因子：
3.6
通讯作者：
Klotz, KN
中科院分区：
文献类型：
--
作者：
Hoffmann, C;Leitz, MR;Klotz, KN
尽管许多β（1）-受体拮抗剂和β（2）-受体激动剂已用于药物治疗多年，但它们对所有三种已知的β-肾上腺素能受体亚型的药理学特性并不总是很好地表征。因此，本研究的目的是提供激动剂的比较结合特性（肾上腺素、去甲肾上腺素、异丙肾上腺素、非诺特罗、沙丁胺醇、沙美特罗、特布他林、福莫特罗、溴沙特罗）和拮抗剂（普萘洛尔，阿普洛尔，阿替洛尔，美托洛尔，比索洛尔，卡维地洛，吲哚洛尔，BRL 37344，CGP 20712，SR 59230 A，CGP 12177，ICI 118551）在相同细胞背景下对所有三种人类β-肾上腺素能受体亚型产生作用。我们产生了稳定表达三种β-肾上腺素能受体亚型的中国仓鼠卵巢（CHO）细胞，表达水平相当。我们使用非选择性拮抗剂I-125-cyanopindolol作为放射性配体，对这些受体亚型进行了表征，并分析了常规使用药物以及竞争结合研究中实验化合物的亲和力。此外，我们分析了β-受体介导的腺苷酸环化酶的活性，从这些细胞系的分离膜。我们的实验结果表明，所有化合物对三种β受体亚型表现出不同的选择性和活性模式。特别是，鉴定了许多β（2）-或β（3）-受体激动剂，它们是其他亚型的反向激动剂。此外，还发现了对β（2）和β（3）受体具有激动活性的β（1）受体拮抗剂，这些对不同亚型的激动、拮抗和反向激动作用的特定混合物可能对相应化合物的治疗用途具有重要意义。
Although many beta(1)-receptor antagonists and beta(2)-receptor agonists have been used in pharmacotherapy for many years their pharmacological properties at all three known subtypes of beta-adrenergic receptors are not always well characterized. The aim of this study was, therefore, to provide comparative binding characteristics of agonists (epinephrine, norepinephrine, isoproterenol, fenoterol, salbutamol, salmeterol, terbutalin, formoterol, broxaterol) and antagonists (propranolol, alprenolol, atenolol, metoprolol, bisoprolol, carvedilol, pindolol, BRL 37344, CGP 20712, SR 59230A, CGP 12177, ICI 118551) at all three subtypes of human beta-adrenergic receptors in an identical cellular background. We generated Chinese hamster ovary (CHO) cells stably expressing the three beta-adrenergic receptor subtypes at comparable levels. We characterized these receptor subtypes and analyzed the affinity of routinely used drugs as well as experimental compounds in competition binding studies, using the non-selective antagonist I-125-cyanopindolol as a radioligand. Furthermore, we analyzed the beta-receptor-mediated adenylyl cyclase activity in isolated membranes from these cell lines. The results from our experiments show that all compounds exhibit distinct patterns of selectivity and activity at the three beta-receptor subtypes. In particular, a number of beta(2)- or beta(3)-receptor agonists that are inverse agonists at the other subtypes were identified. In addition, beta(1)-receptor antagonists with agonistic activity at beta(2)- and beta(3)-receptors were found. These specific mixtures of agonism, antagonism, and inverse agonism at different subtypes may have important implications for the therapeutic use of the respective compounds.