title: Experimental studies on guanosine 3',5'-cyclic monophosphate levels and airway responsiveness of the novel phosphodiesterase type 5 inhibitor SR 265579 in guinea-pigs.
abstract: The effect of 1-cyclopentyl-3-ethyl-6-(3-ethoxypyrid-4-yl)- 1H-pyrazolo[3,4-d]pyrimidin-4-one (SR 265579), a potent inhibitor of guanosine 3',5'-cyclic monophosphate (cyclic GMP) phosphodiesterase (PDE5), was examined regarding its specificity toward the other cyclic nucleotide phosphodiesterases, the effect on cyclic nucleotide levels and the bronchodilatory activity, both in vitro and in vivo in guinea-pigs. The effects were compared to those obtained with zaprinast (CAS 37762-06-4), a known PDE5 inhibitor. Anion-exchange chromatography of the soluble fraction of guinea-pig homogenates revealed 5 peaks which corresponded to PDE1, PDE2, PDE3, PDE4 and PDE5. SR 265579 produced a potent and competitive inhibition, with respect to cyclic GMP, of PDE5 with a Ki of 6.4 nmol/l. The compound was 25 fold more potent than zaprinast and demonstrated selectivity toward PDE5. The selectivity index was 14 and 33 with respect to PDE4 and 3, respectively. PDE1 and 2 were only inhibited at considerably higher concentrations. SR 265579 specifically increased the intracellular cyclic GMP levels in guinea-pig tracheal epithelial cells (EC50 = 117 nmol/l). Moreover, in the guinea pig, plasma cyclic GMP levels were significantly increased after the intravenous or oral administration of doses as low as 1 mg/kg. Isolated guinea-pig trachea were relaxed by the addition of SR 265579 as evaluated by measuring either spontaneous tone or relaxation of histamine and acetylcholine-precontracted preparations. PD2 values were of 7.64, 6.52 and 5.25, respectively. In vivo, after i.v. administration, bronchodilatory activity was demonstrated in an artificially-ventilated guinea-pig histamine-induced bronchospasm model with an ED50 of 0.63 mg/kg. In all experiments, SR-265579 was proved to be more active than zaprinast. These results demonstrate that SR 265579 is an orally active, potent and specific inhibitor of PDE5.
authors.affiliation: Biological Research Department, Chinoin Pharmaceutical and Chemicals Works, Budapest, Hungary. Zoltan.Kapui@Sanofi.com
authors.forename: Z
authors.initials: Z
authors.lastname: Kapui
authors.forename: P
authors.initials: P
authors.lastname: Schaeffer
authors.forename: E G
authors.initials: EG
authors.lastname: Mikus
authors.forename: E
authors.initials: E
authors.lastname: Boronkay
authors.forename: J
authors.initials: J
authors.lastname: Gyürky
authors.forename: J M
authors.initials: JM
authors.lastname: Herbert
authors.forename: M
authors.initials: M
authors.lastname: Pascal
mesh_terms: D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000818:Animals; D016084:Bronchoconstriction / Q000187:drug effects; D006152:Cyclic GMP / Q000097:blood / Q000378:metabolism*; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D006168:Guinea Pigs; D006632:Histamine / Q000494:pharmacology; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D009129:Muscle Tonus / Q000187:drug effects; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D010972:Platelet Activating Factor / Q000494:pharmacology; D012137:Respiratory System / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D014132:Trachea / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
chemical_list: D007527:Isoenzymes; D010726:Phosphodiesterase Inhibitors; D010972:Platelet Activating Factor; D006632:Histamine; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D006152:Cyclic GMP