In vitro and in vivo pharmacological characterization of the neuropeptide s receptor antagonist [D-Cys(tBu)5]neuropeptide S.
In vitro and in vivo pharmacological characterization of the neuropeptide s receptor antagonist [D-Cys(tBu)5]neuropeptide S.
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DOI：
10.1124/jpet.108.143867
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发表时间：
2009-02
期刊：
影响因子：
--
通讯作者：
Calò G
中科院分区：
文献类型：
--
作者：
Camarda V;Rizzi A;Ruzza C;Zucchini S;Marzola G;Marzola E;Guerrini R;Salvadori S;Reinscheid RK;Regoli D;Calò G
神经肽S （NPS）被鉴定为孤儿受体的内源性配体，现在称为NPSR。在对NPS Gly5进行的结构-活性研究框架中，鉴定了NPSR配体[D-Cys(tBu)5]NPS。[D-Cys(tBu)5]高达100 μM的NPS对稳定表达小鼠NPSR （HEK293mNPSR）的HEK293细胞的钙动员没有刺激作用，但该肽以浓度依赖的方式抑制了10和100 nM NPS （pK 6.62）引起的刺激作用。在Schild分析实验中[D-Cys(tBu)5]NPS （0.1 ~ 100 μM）产生了浓度依赖且浓度响应曲线向NPS平行右移，pA2值为6.44。10 μM [D-Cys(tBu)5]NPS对7个与NPSR无关的g蛋白偶联受体的信号传导没有影响。在小鼠翻正反射（RR）恢复试验中，0.1 nmol脑室内给予NPS可降低地西泮15 mg/kg小鼠翻正反射丧失的百分比和睡眠时间。[D-Cys(tBu)5]NPS （1-10 nmol）本身无活性，但能剂量依赖性地拮抗NPS的促觉醒作用。最后，npsr缺陷小鼠对地西泮的催眠效果和野生型小鼠一样敏感。然而，在野生型小鼠中可以检测到1 nmol NPS的唤醒作用，而在突变小鼠中则没有。综上所述，[D-Cys(tBu)5]NPS在体外和体内都是一种纯的、选择性的NPSR拮抗剂，但效力适中。此外，利用该工具和受体敲除小鼠研究，我们证明了NPS的唤醒促进作用是由于NPSR蛋白的选择性激活。
Neuropeptide S (NPS) was identified as the endogenous ligand of an orphan receptor now referred to as NPSR. In the frame of a structure-activity study performed on NPS Gly5, the NPSR ligand [D-Cys(tBu)5]NPS was identified. [D-Cys(tBu)5]NPS up to 100 μM did not stimulate calcium mobilization in HEK293 cells stably expressing the mouse NPSR (HEK293mNPSR), however, the peptide inhibited in a concentration dependent manner the stimulatory effects elicited by 10 and 100 nM NPS (pK 6.62). In Schild analysis experiments [D-Cys(tBu)5]NPS (0.1 - 100 μM) produced a concentration dependent and parallel rightward shift of the concentration response curve to NPS showing a pA2 value of 6.44. 10 μM [D-Cys(tBu)5]NPS did not affect signalling at seven NPSR unrelated G-protein coupled receptors. In the mouse righting reflex (RR) recovery test, NPS given at 0.1 nmole intracerebroventricularly reduced the percent of animals losing the RR in response to diazepam 15 mg/kg and their sleeping time. [D-Cys(tBu)5]NPS (1-10 nmoles) was inactive per se but dose dependently antagonized the arousal-promoting action of NPS. Finally, NPSR-deficient mice were similarly sensitive to the hypnotic effects of diazepam as their wild-type littermates. However, the arousal promoting action of 1 nmole NPS could be detected in wild-type but not in mutant mice. In conclusion, [D-Cys(tBu)5]NPS behaves both in vitro and in vivo as a pure and selective NPSR antagonist, but with moderate potency. Moreover, using this tool together with receptor knockout mice studies we demonstrated that the arousal-promoting action of NPS is due to the selective activation of the NPSR protein.
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影响因子：
3.4
作者：
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通讯作者：
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影响因子：
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作者：
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作者：
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通讯作者：
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