Nodin Christina, Vauquelin Georges, von Mentzer Bengt
Preclinical Research & Development, AstraZeneca Mölndal, 43183 Mölndal, Sweden.
Biochem Pharmacol. 2005 Apr 15;69(8):1235-40. doi: 10.1016/j.bcp.2005.01.008.
The present study reveals that cystein2,7 ethyl-amidealphaCGRP (Cys2,7EtalphaCGRP), an advertised calcitonin gene-related peptide 2 (CGRP2) receptor subtype-selective agonist, is also a potent agonist for the calcitonin gene-related peptide 1 (CGRP1) receptors natively expressed in the SK-N-MC human neuroblastoma cell line. Cys2,7EtalphaCGRP and alpha calcitonin gene-related peptide (alphaCGRP) promote cyclic AMP accumulation in intact SK-N-MC cells to the same extent with EC50 of 1.6+/-0.2 and 0.4+/-0.08 nM, respectively. The antagonist alpha calcitonin gene-related peptide-8-37 (alphaCGRP-(8-37)) produces a concentration-dependent rightward shift of the alphaCGRP- and Cys2,7EtalphaCGRP concentration-response curves with KB-values (71+/-33 and 47+/-21 nM, respectively). The competitive antagonism by alphaCGRP-(8-37) and the similar KB-values suggests that alphaCGRP and Cys2,7EtalphaCGRP stimulate the same receptor. In competition binding studies with [125I]-alphaCGRP on SK-N-MC cell membranes, Cys2,7EtalphaCGRP and alphaCGRP-(8-37) display high affinity for the majority of the binding sites with Ki-values of 0.030+/-0.013 and 0.60+/-0.013 nM, respectively. The present findings are at odds with the proclaimed utilization of Cys2,7EtalphaCGRP as a CGRP2 receptor-selective pharmacological tool. Differences between the agonistic profile of this ligand in this and other experimental systems might be species--or even cell type--dependent.
本研究表明,宣传为降钙素基因相关肽2(CGRP2)受体亚型选择性激动剂的半胱氨酸2,7乙酰胺αCGRP(Cys2,7EtαCGRP),也是在SK-N-MC人神经母细胞瘤细胞系中天然表达的降钙素基因相关肽1(CGRP1)受体的强效激动剂。Cys2,7EtαCGRP和α降钙素基因相关肽(αCGRP)在完整的SK-N-MC细胞中促进环磷酸腺苷积累的程度相同,EC50分别为1.6±0.2和0.4±0.08 nM。拮抗剂α降钙素基因相关肽-8-37(αCGRP-(8-37))使αCGRP和Cys2,7EtαCGRP浓度-反应曲线产生浓度依赖性右移,KB值分别为(71±33和47±21 nM)。αCGRP-(8-37)的竞争性拮抗作用和相似的KB值表明αCGRP和Cys2,7EtαCGRP刺激相同的受体。在用[125I]-αCGRP对SK-N-MC细胞膜进行的竞争结合研究中,Cys2,7EtαCGRP和αCGRP-(8-37)对大多数结合位点显示出高亲和力,Ki值分别为0.030±0.013和0.60±0.013 nM。本研究结果与宣称将Cys2,7EtαCGRP用作CGRP2受体选择性药理学工具的说法不一致。该配体在本实验系统和其他实验系统中的激动特性差异可能与物种甚至细胞类型有关。