Identification of human dopamine D1-like receptor agonist using a cell-based functional assay
Source: Acta Pharmacologica Sinica, Volume 26, Number 10, October 2005 , pp. 1181-1186(6)
Abstract:Aim: To establish a cell-based assay to screen human dopamine D1 and D5 recep-tor agonists against compounds from a natural product compound library. Methods: Synthetic responsive elements 6×cAMP response elements (CRE) and a mini promoter containing a TATA box were inserted into the pGL3 basic vector to generate the reporter gene construct pCRE/TA/Luci. CHO cells were co-trans-fected with the reporter gene construct and human D1 or D5 receptor cDNA in mammalian expression vectors. Stable cell lines were established for agonist screening. A natural product compound library from over 300 herbs has been established. The extracts from these herbs were used for human D1 and D5 receptor agonist screenings. Results: A number of extracts were identified that activated both D1 and D5 receptors. One of the herb extracts, SBG492, demon-strated distinct pharmacological characteristics with human D1 and D5 receptors. The EC50 values of SBG492 were 342.7 g/mL for the D1 receptor and 31.7 g/mL for the D5 receptor. Conclusion: We have established a cell-based assay for high-throughput drug screening to identify D1-like receptor agonists from natural products. Several extracts that can active D1-like receptors were discovered. These compounds could be useful tools for studies on the functions of these receptors in the brain and could potentially be developed into therapeutic drugs for the treatment of central nervous system diseases.
Document Type: Research article
Affiliations: 1: Shanghai Institute of Brain Functional Genomics and Key Lab of Brain Functional Genomics, Ministry of Education, Science and Technology Commission of Shanghai Municipality, East China Normal University, Shanghai 200062; 2: College of Bioengineering, Chongqing University, Chongqing 400044, China
Publication date: 2005-10-01