The Protective Effect of DL0805 Derivatives on Pulmonary Artery Cells and the Underlying Mechanisms Study.
BACKGROUND
Pulmonary hypertension (PH) is a severe disease characterized by a progressive increase in pulmonary vascular resistance, initially due to abnormal pulmonary vasoconstriction in response to endothelial and smooth muscle cells injury. The discovery of new chemical entities having a protective effect on pulmonary artery cells could be meaningful for the treatment of PH.


METHODS
We evaluated the protective effect of DL0805 derivatives (DL0805-1 and DL0805-2) on pulmonary artery vascular cells, including human pulmonary artery endothelial cells (HPAECs) and human pulmonary artery smooth muscle cells (HPASMCs). DL0805 derivatives are novel ROCKs (Rho-associated coiled-coil forming protein serine/threonine kinases) inhibitors. Treatment of HPAECs with DL0805-2 (10 μM) cultured under a hypoxic environment could significantly reduce the proliferation of cells. Meanwhile, the compounds inhibited the production of reactive oxygen species (ROS) in HPAECs at every dose tested.


RESULTS
A Western Blot experiment showed that the protective effect of DL0805 derivatives might result from the down-regulation of RhoA (Ras homolog gene family, member A) expression and the inhibition of ROCKs activity. In addition, the compounds inhibited the proliferation of HPASMCs induced by fetal bovine serum (FBS) or platelet derived growth factor BB (PDGF-BB), and suppressed the F-actin remodeling induced by endothelin.


CONCLUSIONS
The preliminary results from an immunofluorescence assay showed that DL0805 derivatives inhibited the activity of ROCKs in HPASMCs. The above mentioned results indicated that DL0805 derivatives have a protective effect on pulmonary artery cells, and the underlying mechanisms might be the result of inhibition of RhoA/ROCK signaling pathway.
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College: Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Beijing 100050. China.
Tianyi
T
Yuan
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College: Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Beijing 100050. China.
Huifang
H
Zhang
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College: Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Beijing 100050. China.
Yucai
Y
Chen
Beijing Key Laboratory of Active Substance Discovery and Druggability Evaluation, Beijing 100050. China.
Xiaozhen
X
Jiao
Beijing Key Laboratory of Active Substance Discovery and Druggability Evaluation, Beijing 100050. China.
Ping
P
Xie
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, National center for pharmaceutical screening, P.O. Box: 100050, Beijing. China.
Lianhua
L
Fang
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, National center for pharmaceutical screening, P.O. Box: 100050, Beijing. China.
Guanhua
G
Du
D000077214:Becaplermin; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D015536:Down-Regulation / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D006976:Hypertension, Pulmonary / Q000188:drug therapy; D007191:Indazoles / Q000737:chemistry / Q000494:pharmacology*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000378:metabolism; D009570:Nitriles / Q000737:chemistry / Q000494:pharmacology*; D020574:Proto-Oncogene Proteins c-sis / Q000378:metabolism; D011651:Pulmonary Artery / Q000166:cytology / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D054460:rho-Associated Kinases / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
C575013:5-nitro-1(2)H-indazole-3-carbonitrile; D007191:Indazoles; C000598341:N-(1H-indazol-5-yl)-1-(4-methylbenzyl)pyrrolidine-3-carboxamide; D009570:Nitriles; D020574:Proto-Oncogene Proteins c-sis; D017382:Reactive Oxygen Species; D000077214:Becaplermin; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein
DL0805 derivatives; HPAEC; HPASMC; ROCKs; hypoxia; proliferation