Characterisation of the Binding of [3H]WAY‐100635, a Novel 5‐Hydroxytryptamine1A Receptor Antagonist, to Rat Brain
Characterisation of the Binding of [3H]WAY‐100635, a Novel 5‐Hydroxytryptamine1A Receptor Antagonist, to Rat Brain
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新型 5-羟色胺 1A 受体拮抗剂 [3H]WAY-100635 与大鼠大脑结合的表征
DOI：
10.1046/j.1471-4159.1995.64062716.x
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发表时间：
1995
影响因子：
4.7
通讯作者：
M. Minchin
中科院分区：
文献类型：
--
作者：
X. Khawaja;N. Evans;Y. Reilly;C. Ennis;M. Minchin
摘要：[3H]WAY-100635 {N-[2-[4-(2-[O-甲基-3H]甲氧基苯基)-1-哌嗪基]乙基]-N-(2-吡啶基)环己烷甲酰胺三盐酸盐}与大鼠海马膜制剂的特异性结合是时间、温度和组织浓度。 依赖。 [3H]WAY-100635 缔合率 (k+1 = 0.069 ± 0.015 nM−1 min−1) 和解离率 (k−1 = 0.023 ± 0.001 min−1) 遵循单指数动力学。 [3H]WAY-100635 的饱和结合等温线表现出单一类别的识别位点，亲和力为 0.37 ± 0.051 nM，最大结合能力 (Bmax) 为 312 ± 12 fmol/mg 蛋白质。与 8-羟基-2-(二-正-[3H]-丙氨基)四氢萘 ([3H]8-OH-DPAT) 相比，[3H]WAY-100635 标记的最大结合位点数量高出约 36%。二价阳离子 CaCl2（2.5 倍；p < 0.02）和 MnCl2（3.6 倍；p < 0.05）显着降低 [3H]WAY-100635 的结合亲和力，但对 Bmax 没有影响。鸟苷酸未能影响 [3H]WAY-100635 与 5-HT1A 受体结合的 KD 和 Bmax 参数。 [3H]WAY-100635 的药理学结合特征与 [3H]8-OH-DPAT 密切相关，这与大鼠海马中 5-羟色胺1A (5-HT1A) 位点的标记一致。 [3H]WAY-100635 与 5-HT1A 激动剂和部分激动剂的竞争曲线最好分解为高亲和力和低亲和力结合成分，而拮抗剂最好通过单位点结合模型来描述。在 50 µM 鸟苷 5'-O-(3-硫代三磷酸) (GTPγS) 存在下，拮抗剂的竞争曲线保持不变，而激动剂和部分激动剂曲线向右移动，反映了 G 蛋白偶联对激动剂与拮抗剂与 5-HT1A 受体结合的影响。然而，在 GTPγS 存在的情况下，残留的 (16 ± 2%) 高亲和力激动剂结合成分仍然很明显，表明存在 GTP 不敏感位点。
Abstract: The specific binding of [3H]WAY‐100635 {N‐[2‐[4‐(2‐[O‐methyl‐3H]methoxyphenyl)‐1‐piperazinyl]ethyl]‐N‐(2‐pyridinyl)cyclohexane carboxamide trihydrochloride} to rat hippocampal membrane preparations was time, temperature, and tissue concentration dependent. The rates of [3H]WAY‐100635 association (k+1 = 0.069 ± 0.015 nM−1 min−1) and dissociation (k−1 = 0.023 ± 0.001 min−1) followed monoexponential kinetics. Saturation binding isotherms of [3H]WAY‐100635 exhibited a single class of recognition site with an affinity of 0.37 ± 0.051 nM and a maximal binding capacity (Bmax) of 312 ± 12 fmol/mg of protein. The maximal number of binding sites labelled by [3H]WAY‐100635 was ∼36% higher compared with that of 8‐hydroxy‐2‐(di‐n‐[3H]‐propylamino)tetralin ([3H]8‐OH‐DPAT). The binding affinity of [3H]WAY‐100635 was significantly lowered by the divalent cations CaCl2 (2.5‐fold; p < 0.02) and MnCl2 (3.6‐fold; p < 0.05), with no effect on Bmax. Guanyl nucleotides failed to influence the KD and Bmax parameters of [3H]WAY‐100635 binding to 5‐HT1A receptors. The pharmacological binding profile of [3H]WAY‐100635 was closely correlated with that of [3H]8‐OH‐DPAT, which is consistent with the labelling of 5‐hydroxytryptamine1A (5‐HT1A) sites in rat hippocampus. [3H]WAY‐100635 competition curves with 5‐HT1A agonists and partial agonists were best resolved into high‐ and low‐affinity binding components, whereas antagonists were best described by a one‐site binding model. In the presence of 50 µM guanosine 5′‐O‐(3‐thiotriphosphate) (GTPγS), competition curves for the antagonists remained unaltered, whereas the agonist and partial agonist curves were shifted to the right, reflecting an influence of G protein coupling on agonist versus antagonist binding to the 5‐HT1A receptor. However, a residual (16 ± 2%) high‐affinity agonist binding component was still apparent in the presence of GTPγS, indicating the existence of GTP‐insensitive sites.
用于治疗焦虑症和抑郁症的血清素特异性药物。
影响因子：
10.5
作者：
Charney,DS;Krystal,JH;Delgado,PL;Heninger,GR
通讯作者：
Heninger,GR
DOI：
10.1016/s0021-9258(19)39437-2
发表时间：
1990-04
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Paul R. Albert;Qun-Yong Zhou;H. V. Tol;J. Bunzow;O. Civelli
通讯作者：
Paul R. Albert;Qun-Yong Zhou;H. V. Tol;J. Bunzow;O. Civelli
咪唑啉 UK-14, 304（一种假定的全 α2-肾上腺素受体激动剂）与大鼠大脑皮层膜的结合。
影响因子：
6.1
作者：
Loftus,DJ;Stolk,JM;U'Prichard,DC
通讯作者：
U'Prichard,DC