Adham N, Bard J A, Zgombick J M, Durkin M M, Kucharewicz S, Weinshank R L, Branchek T A
Synaptic Pharmaceutical Corporation, Paramus, NJ 07652, USA.
Neuropharmacology. 1997 Apr-May;36(4-5):569-76. doi: 10.1016/s0028-3908(97)00020-8.
The anti-migraine compound, sumatriptan, has been shown to have substantial affinity for the cloned human 5-HT1F receptor suggesting that, in addition to 5-HT1B/5-HT1D receptor subtypes, the 5-HT1F receptor may be a therapeutic target for the treatment of migraine. Several investigators have used the guinea pig plasma extravasation model to evaluate potential anti-migraine drugs. Since species differences in the pharmacology of serotonin receptors are well known, we compared the pharmacological profiles of the cloned human and guinea pig 5-HT1F receptors in order to validate the usefulness of the in vivo model in predicting anti-migraine activity of compounds targeted for humans. We have cloned the guinea pig 5-HT1F by homology to the human 5-HT1F receptor and evaluated its pharmacological profile using radioligand binding assays. The cloned guinea pig 5-HT1F gene exhibited 94% amino acid identity to the corresponding human homolog. High affinity (Kd approximately 10 nM) [3H]5-HT binding was detected to membranes obtained from Cos-7 cells transiently expressing the guinea pig 5-HT1F receptor. The cloned guinea pig receptor displayed typical 5-HT1F receptor pharmacology with the following rank order of binding affinities: 5-HT > sumatriptan > 1-NP = DHE > alpha-methyl 5-HT > metergoline > methiothepin > 5-CT. The pharmacological profiles of the cloned guinea pig and human 5-HT1F receptors were very similar as reflected by the high correlation (r2 = 0.72, slope = 0.76) observed between the binding affinities of compounds for these two species homologs. In situ hybridization studies in guinea pig tissue revealed 5-HT1F receptor mRNA expression in the neurons of the trigeminal ganglion, suggesting that the 5-HT1F receptor may play a role in the presynaptic inhibition of neuropeptide release at the level of the intracranial vasculature, thereby blocking the development of neurogenic inflammation. Dorsal root ganglion cells also moderately expressed the 5-HT1F transcripts. The localization of the 5-HT1F receptor to areas involved in the mediation and transfer of nociceptive information implies a role for this receptor in pain processing. These findings indicate that a selective 5-HT1F agonist may be a novel approach to treat migraine.
抗偏头痛化合物舒马曲坦已被证明对克隆的人5-HT1F受体具有显著亲和力,这表明除了5-HT1B/5-HT1D受体亚型外,5-HT1F受体可能是治疗偏头痛的一个治疗靶点。几位研究人员已使用豚鼠血浆外渗模型来评估潜在的抗偏头痛药物。由于血清素受体药理学的种属差异是众所周知的,我们比较了克隆的人5-HT1F受体和豚鼠5-HT1F受体的药理学特征,以验证体内模型在预测针对人类的化合物的抗偏头痛活性方面的有用性。我们通过与人5-HT1F受体的同源性克隆了豚鼠5-HT1F受体,并使用放射性配体结合试验评估了其药理学特征。克隆的豚鼠5-HT1F基因与相应的人同源物具有94%的氨基酸同一性。在从瞬时表达豚鼠5-HT1F受体的Cos-7细胞获得的膜上检测到高亲和力(Kd约为10 nM)的[3H]5-HT结合。克隆的豚鼠受体表现出典型的5-HT1F受体药理学特征,其结合亲和力的顺序如下:5-HT>舒马曲坦>1-NP = DHE>α-甲基5-HT>美替拉酮>甲硫噻平>5-CT。克隆的豚鼠和人5-HT1F受体的药理学特征非常相似,这一点通过观察到的这两种种属同源物的化合物结合亲和力之间的高度相关性(r2 = 0.72,斜率 = 0.76)得到反映。豚鼠组织中的原位杂交研究揭示了三叉神经节神经元中5-HT1F受体mRNA的表达,这表明5-HT1F受体可能在颅内血管水平的神经肽释放的突触前抑制中发挥作用,从而阻断神经源性炎症的发展。背根神经节细胞也适度表达5-HT1F转录本。5-HT1F受体在参与伤害性信息介导和传递的区域的定位意味着该受体在疼痛处理中发挥作用。这些发现表明,选择性5-HT1F激动剂可能是治疗偏头痛的一种新方法。