Identification of human dopamine receptors agonists from Chinese herbs.
OBJECTIVE
To find human dopamine receptors, especially D1-like receptor specific agonists from Chinese herbs as potential antihypertension drug leads.


METHODS
Two D1-like receptor cell lines carrying a beta-lactamase reporter gene, and a D2 receptor cell line coexpressing a promiscuous G protein G15 were constructed using HEK293 cells. A natural compound library made from fractionated samples of herbal extracts was used for high-throughput screening (HTS) against one of the cell lines, HEK/D5R/CRE-blax. The interested hits were evaluated for their activities against various dopamine receptors.


RESULTS
Fourteen hits were identified from primary screening, of which 2 of the better hit samples, HD0522 and HD0059, were selected for further material and activity analysis, and to obtain 2 compounds that appeared as 2 single peaks in HPLC, HD0522H01 and HD0059H01. HD0059H01 could activate D1, D2, and D5 receptors, with EC(50 ) values of 2.28 microg/mL, 0.85 microg/mL, and 1.41 microg/mL, respectively. HD0522H01 could only activate D1R and D5R with EC(50 ) values of 2.95 microg/mL and 8.38 microg/mL.


CONCLUSIONS
We established cellbased assays for 3 different human dopamine receptors and identified specific agonists HD0522H01 and HD0059H01 through HTS. The specific agonist to D1-like receptors, HD0522H01, may become a new natural product-based drug lead for antihypertension treatment.
Zhang
Yi-Lin
YL
Graduate School of the Chinese Academy of Sciences, Beijing 100039, China.
Zhang
Hai-Qing
HQ
Liu
Xiao-Yu
XY
Hua
Shi-Neng
SN
Zhou
Lu-Bing
LB
Yu
Jun
J
Tan
Xue-Hai
XH
D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000302:isolation & purification / Q000494:pharmacology*; D016147:Genes, Tumor Suppressor; D006801:Humans; D010946:Plants, Medicinal / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D017447:Receptors, Dopamine D1 / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D017448:Receptors, Dopamine D2 / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D050641:Receptors, Dopamine D5 / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism
D018797:Cell Cycle Proteins; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D004365:Drugs, Chinese Herbal; C473448:RBM5 protein, human; D016601:RNA-Binding Proteins; D017447:Receptors, Dopamine D1; D017448:Receptors, Dopamine D2; D025521:Tumor Suppressor Proteins; D050641:Receptors, Dopamine D5; D001618:beta-Lactamases