CHARACTERIZATION OF A NOVEL POTENT AND SPECIFIC INHIBITOR OF TYPE-V PHOSPHODIESTERASE

被引:53
作者
COSTE, H [1 ]
GRONDIN, P [1 ]
机构
[1] CTR RECH, GLAXO LABS, F-91951 LES ULIS, FRANCE
关键词
PHOSPHODIESTERASE; CGMP; SMOOTH MUSCLE CELL; EGRESSION;
D O I
10.1016/0006-2952(95)02031-4
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Guanosine cyclic 3':5'-monophosphate (cGMP) plays a crucial role in regulating vascular smooth muscle contractile state. In rat aortic smooth muscle cells (RSMC) three isozymes of phosphodiesterase (PDE) may be involved in the degradation of cGMP, namely PDE I, PDE III, and PDE V. To study the effective contribution of PDE V to the control of intracellular cGMP levels, a specific and potent PDE V inhibitor 1,3-dimethyl-6-(2-propoxy-5-methanesulfonylamidophenyl)pyrazolo[3,4d]-pyrimidin-4-(5H)-one (DMPPO) was synthesized. DMPPO is a competitive inhibitor with respect to cGMP (K-i = 3 nM) and displayed high selectivity for PDE V as compared to other PDE isozymes. DMPPO strongly potentiated the cGMP response of atrial natriuretic peptide- or sodium nitroprusside-treated RSMC (EC(50) = 0.5 mu M). In addition, similar intracellular cGMP levels were obtained in the presence of a saturating concentration of DMPPO or 3-isobutyl-1-methylxanthine, a nonspecific PDE inhibitor, suggesting that cGMP is almost exclusively hydrolyzed by PDE V in RSMC. Stimulation of RSMC with atrial natriuretic factor resulted in accumulation of cGMP in the extracellular media. This egression was shown to be proportional to the intracellular level of cGMP and a first-order rate constant of 0.04 min(-1) was determined for the egression process. DMPPO did not interfere with the efflux and allowed us to show that intracellular cGMP levels are mainly controlled by PDE V, rather than by egression in RSMC. DMPPO is, therefore, a useful tool for determining the role of PDE V in the control of cGMP levels in living cells and tissues.
引用
收藏
页码:1577 / 1585
页数:9
相关论文
共 47 条
[1]  
ANANDSRIVASTAVA MB, 1993, PHARMACOL REV, V45, P455
[2]  
BARBER R, 1980, J CYCLIC NUCL PROT, V6, P3
[3]  
BARBER R, 1983, ADV CYCL NUCL RES<D>, V15, P119
[4]  
BARBER R, 1981, MOL PHARMACOL, V19, P38
[5]  
BEAVO J, 1990, MOL PHARM CELL REGUL, V2, P3
[6]   A FAMILY OF HUMAN PHOSPHODIESTERASES HOMOLOGOUS TO THE DUNCE LEARNING AND MEMORY GENE-PRODUCT OF DROSOPHILA-MELANOGASTER ARE POTENTIAL TARGETS FOR ANTIDEPRESSANT DRUGS [J].
BOLGER, G ;
MICHAELI, T ;
MARTINS, T ;
STJOHN, T ;
STEINER, B ;
RODGERS, L ;
RIGGS, M ;
WIGLER, M ;
FERGUSON, K .
MOLECULAR AND CELLULAR BIOLOGY, 1993, 13 (10) :6558-6571
[7]   ANTIALLERGIC ACTIVITY OF 2-PHENYL-8-AZAPURIN-6-ONES [J].
BROUGHTON, BJ ;
CHAPLEN, P ;
KNOWLES, P ;
LUNT, E ;
MARSHALL, SM ;
PAIN, DL ;
WOOLDRIDGE, KRH .
JOURNAL OF MEDICINAL CHEMISTRY, 1975, 18 (11) :1117-1122
[8]  
BRUNTON LL, 1988, METHOD ENZYMOL, V159, P83
[9]   THE CATALYTIC SUBUNIT OF PROTEIN KINASE-A TRIGGERS ACTIVATION OF THE TYPE-V CYCLIC GMP-SPECIFIC PHOSPHODIESTERASE FROM GUINEA-PIG LUNG [J].
BURNS, F ;
RODGER, IW ;
PYNE, NJ .
BIOCHEMICAL JOURNAL, 1992, 283 :487-491
[10]  
CHAMLEY JH, 1977, CELL TISSUE RES, V177, P503