BAY 41-2272 [5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3-yl]pyrimidin-4-ylamine]-induced dilation in ovine pulmonary artery: role of sodium pump.

The mechanisms of relaxation to nitric oxide (NO)-independent soluble guanylyl cyclase (sGC) activator BAY 41-2272 [5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3-yl]pyrimidin-4-ylamine] were investigated in isolated ovine pulmonary artery. BAY 41-2272 (1 nM-10 microM) produced concentration-dependent relaxation of endothelium-denuded pulmonary artery rings (pD2 = 6.82 +/- 0.16; Emax = 92.30 +/- 2.31%; n = 8), precontracted with 1 microM 5-hydroxytryptamine (serotonin). 1-H-[1,2,4]Oxadiazole[4,3-a]quinoxalin-1-one (ODQ; 10 microM), an inhibitor of sGC, partially inhibited (Emax = 57.10 +/- 3.10%; n = 6) the relaxation response of BAY 41-2272. In comparison with ODQ, sodium pump inhibitor ouabain (1 microM) produced a greater decrease in the vasodilator response of BAY 41-2272 (Emax = 20.17 +/- 4.55%; n = 6). K+-free solution also attenuated (Emax = 39.97 +/- 3.52%; n = 6) BAY 41-2272-induced relaxation. ODQ (10 microM) plus 1 microM ouabain abolished the relaxant response of BAY 41-2272 (Emax = 12.09 +/- 3.76%, n = 6 versus vehicle control dimethyl sulfoxide; Emax = 15.83 +/- 1.72%, n = 6). KT-5823 [1-oxo-9.12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-I][1,6]benzodiazocine-10-carboxylic acid methyl ester (2 microM), a specific inhibitor of protein kinase G had no effect on 10 microM ODQ-insensitive relaxation evoked by BAY 41-2272. BAY 41-2272 (10 microM) inhibited Ca2+-induced contractions in K+-depolarized preparations. BAY 41-2272 (10 microM) caused about a 14-fold increase in the intracellular cGMP over the basal level, which was completely inhibited by 10 microM ODQ. BAY 41-2272 (0.1, 1.0, and 10 microM) significantly (P < 0.05) increased ouabain-sensitive 86Rb uptake in a concentration-dependent manner. BAY 41-2272 (10 microM) also stimulated sarcolemmal Na+-K+-ATPase activity. However, 10 microM ODQ had no significant effect on either basal or BAY 41-2272-stimulated 86Rb uptake/Na+-K+-ATPase activities. In conclusion, this study provides the first evidence of sodium pump stimulation by BAY 41-2272 independent of cGMP as an additional mechanism to sGC activation in relaxation of ovine pulmonary artery.