Drug Discovery Research Based On Natural Products

Chinese herbs, derived from plants in a great measure, are traditional Chinese treasures. The modernization of Chinese traditional medicine is one of the effective approaches to regenerate Chinese traditional medicine with the development of science and technology. The modernization of Chinese traditional medicine is combining Chinese traditional medicine's advantages and characteristics with modern technology, and using modern technological measures to research Chinese herbs and its active mechanism. Chinese herbs with better security, better curative effect, more stable of quality and more convenient would be of great benefits to human.The high-throughput drug screening which is based on Chinese traditional medicine compound library is meaningful for illustrating Chinese herbs active mechanism. In this paper, a functional screening cell model targeting human histamine receptor 3 was established, and special ligands of histamine receptor 3 had been investigated using this cell model by high-throughput drug screening. The high-throughput drug screening was based on the natural compound library that we had constructed.Histamine receptor 3 is a G protein coupled receptor which belongs to Gi subfamily. The promiscuous G protein (G15) was applied to change the signal transduction pathway of histamine receptor 3 from Gi to Gq , and this would be convenient for high-throughput drug screening with calcium assay. HEK293 cell was firstly transfected stably with G15 gene, then was transfected transiently with histamine receptor 3 gene. Clones which expressed G15 protein stably were picked out. The best G15 clone was used as parental cell line, and was transfacted stably with histamine receptor 3 gene. After two weeks'antibiotic screening, a high-throughput drug screening cell model with both histamine receptor 3 and G15 expressed stably was obtained.The stability and speciality of the cell model was tested with histamine which was histamine receptor 3's agonist and clobenpropit antagonist. On the other hand, different ligands dose response curves were detected on the cell model. The result proved that histamine receptor 3 specific agonist and antagonist could induce calcium signal, but...