| Popis: |
Considerable interest has been generated in the potential utility of phosphodiesterase (PDE) IV inhibitors as a novel class of anti-asthmatic agents. Because a detailed understanding of the molecular and biochemical characteristics of any molecular target of interest provides a key ingredient for rational drug design, we cloned a cDNA encoding a PDE IV (hPDE IV) from a human monocyte library and expressed, purified and characterized the recombinant gene product. Purified hPDE IV has kinetic characteristics consistent with native PDE IV isolated from tissue sources. In addition, it is inhibited by rolipram (Ki = 60 nM) and other archetypical PDE IV-selective inhibitors. Purified hPDE IV also contains a high affinity binding site for rolipram (Kd = 2 nM), although the precise relationship between this site and the cAMP catalytic site is not clear. In other studies in which the regulation of PDE IV expression was examined in U937 cells, a human monocytic cell line, prolonged treatment with salbutamol was shown to induce an increase in the activity of PDE IV. This up-regulation of PDE IV activity appears to be mediated by cAMP and occurs at the transcriptional or pretranscriptional level. As a consequence of PDE IV up-regulation, the sensitivity of U937 cells to the inhibitory effects of adenylyl cyclase activators on cell function is greatly diminished. If such regulation of PDE IV occurs in inflammatory cells in vivo, it could have implications for the therapeutic use of beta-adrenoceptor agonists. Specifically, induction of PDE IV activity in asthmatics being treated with beta-adrenoceptor agonists could result in a heterologous desensitization of inflammatory cells to endogenous anti-inflammatory agents (e.g., epinephrine, prostaglandin E2). |
