title: Characterization of the cellular activity of PDE 4 inhibitors using two novel PDE 4 reporter cell lines.
abstract: We report here the generation and pharmacological characterization of two novel PDE 4B1 and PDE 4D3 reporter cell lines. Intracellular cAMP levels are monitored in these cells by a cAMP-sensitive biosensor. We used the recombinant PDE 4B1 and PDE 4D3 reporter cell lines to characterize the cellular effects of various competitive and allosteric PDE 4 inhibitors. In addition, we compared the cellular activity of these PDE 4 inhibitors with the in vitro inhibition of full-length PDE 4D3 and a truncated enzyme comprising the PDE 4D3 catalytic domain. Two different groups of PDE 4 inhibitors could be identified. The first group, including competitive inhibitors like roflumilast, cilomilast and piclamilast, shows similar in vitro activity on full-length and truncated PDE 4D3 and comparably low cellular activity. The second group, including the allosteric inhibitors PMNPQ, D159153, and D159404, shows much better inhibition of full-length versus truncated PDE 4D3. In addition, these compounds show high cellular activity. Our data obtained with the prototype PDE 4 inhibitor rolipram show that rolipram has properties intermediate between the two groups. The results imply that these novel PDE 4 reporter cell lines are well-suited for the characterization of the cellular activity of PDE 4 inhibitors and may also support a better understanding of the complex PDE 4 pharmacology.
authors.affiliation: Lead Discovery Wuppertal, Bayer Pharma AG , Pharma Research Center, Aprather Weg 18a, D-42096 Wuppertal, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Wunder
authors.forename: Ramona
authors.initials: R
authors.lastname: Quednau
authors.forename: Andreas
authors.initials: A
authors.lastname: Geerts
authors.forename: Martina
authors.initials: M
authors.lastname: Barg
authors.forename: Adrian
authors.initials: A
authors.lastname: Tersteegen
mesh_terms: D000631:Aminopyridines / Q000494:pharmacology; D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D016466:CHO Cells; D003412:Cricetulus; D000242:Cyclic AMP / Q000378:metabolism; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4 / Q000378:metabolism*; D054815:Cyclic Nucleotide-Gated Cation Channels / Q000378:metabolism; D003509:Cyclohexanecarboxylic Acids / Q000494:pharmacology; D003521:Cyclopropanes / Q000494:pharmacology; D006801:Humans; D008954:Models, Biological; D009570:Nitriles / Q000494:pharmacology; D058988:Phosphodiesterase 4 Inhibitors / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology; D018342:Receptors, Adrenergic, beta-1 / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000631:Aminopyridines; D001549:Benzamides; D054815:Cyclic Nucleotide-Gated Cation Channels; D003509:Cyclohexanecarboxylic Acids; D003521:Cyclopropanes; D009570:Nitriles; D058988:Phosphodiesterase 4 Inhibitors; D011725:Pyridines; D018342:Receptors, Adrenergic, beta-1; C424423:Roflumilast; C433247:Cilomilast; D000242:Cyclic AMP; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; C087566:piclamilast