Pregenzer J F, Alberts G L, Bock J H, Slightom J L, Im W B
CNS Diseases Research and Molecular Biology, Pharmacia and Upjohn, Inc., Kalamazoo, MI 49001, USA.
Neurosci Lett. 1997 Oct 17;235(3):117-20. doi: 10.1016/s0304-3940(97)00728-3.
The 5-HT1D receptor is a potential target of anti-migraine drugs, and here its genes were cloned from chimpanzee, gorilla and rhesus monkey, via polymerase chain reactions with their genomic DNAs and the primers designed from the 5' and 3' untranslated regions of the human receptor. Direct sequencing of the polymerase chain reaction (PCR) products revealed high degrees of identity between their deduced amino acid sequences (the chimpanzee, gorilla and rhesus monkey) and that of human, differing by two, four and 11 residues, respectively. The binding properties of the receptors, as expressed in human embryonic kidney 293 cells, were compared to those obtained with the human and guinea pig receptors, the latter differing by 33 residues from the human receptor. Standard serotonergic ligands including several indoles, ergots and methiothepin bound all the cloned primate and guinea pig receptors with comparable, low nanomolar affinities, leading to high correlation coefficients among their Ki values. R(+)-8-Hydroxydipropylaminotetralin, on the other hand, bound the human receptor with the affinity higher than those for the primates and guinea pig receptors. This indicates that certain chemical templates may differentiate the molecular divergences among the 5-HT1D receptors of various animal species, and the use of the non-human primates will be beneficial for pharmacological characterizations, more relevant to the human receptor, of future novel ligands for the 5-HT1D receptor, which are potential anti-migraine drugs.
5-羟色胺1D受体是抗偏头痛药物的一个潜在靶点，在此通过聚合酶链反应，利用黑猩猩、大猩猩和恒河猴的基因组DNA以及根据人类受体5'和3'非翻译区设计的引物，克隆了它们的基因。对聚合酶链反应（PCR）产物进行直接测序发现，其推导的氨基酸序列（黑猩猩、大猩猩和恒河猴）与人类的氨基酸序列具有高度同源性，分别相差2个、4个和11个残基。将在人胚肾293细胞中表达的这些受体的结合特性，与人类和豚鼠受体的结合特性进行了比较，豚鼠受体与人类受体相差33个残基。包括几种吲哚、麦角碱和甲硫噻平在内的标准血清素能配体，以相当的低纳摩尔亲和力与所有克隆的灵长类动物和豚鼠受体结合，导致它们的Ki值之间具有高度相关系数。另一方面，R(+)-8-羟基二丙基氨基四氢萘与人类受体的结合亲和力高于灵长类动物和豚鼠受体。这表明某些化学模板可能会区分不同动物物种5-羟色胺1D受体之间的分子差异，使用非人类灵长类动物将有利于对未来新型5-羟色胺1D受体配体（潜在的抗偏头痛药物）进行与人类受体更相关的药理学特性研究。
