Phosphate transport inhibition by KW-3902, an adenosine A1 receptor antagonist, is mediated by cyclic adenosine monophosphate.

We have previously demonstrated that 1,3-dipropyl-8-(3-noradamantyl) xanthine (KW-3902) has an inhibitory effect on phosphate (Pi) transport with no effect on glucose transport in the rat renal proximal tubular cell, similar to that of parathyroid hormone (PTH). In the current studies we investigated the effect of KW-3902, rat PTH (1-34), and 1,3-dipropyl-8-cyclopentylxanthine (DPCPX), another selective adenosine A1 receptor antagonist, on Pi transport and the production of cyclic adenosine monophosphate (cAMP). We then compared these effects of KW-3902 with those of rat PTH in rat renal proximal tubule cells. The results showed that both KW-3902 (30 mumol/L) and rat PTH (1-34, 5 mumol/L) significantly inhibited Pi uptake in proximal cells from a control level of 61 +/- 3 to 19 +/- 3 (a reduction of 69%) and 46 +/- 4 picomoles phosphate/mg protein/min (a reduction of 25%), respectively (P < 0.01). The inhibitory effect of 30 mumol/L KW-3902 alone on Pi transport was more than twice that of 5 mumol/L rat PTH (1-34) alone (P < 0.01). KW-3902 stimulated the production of cAMP in a dose-dependent manner (r = 0.997, P < 0.01). Rat PTH (1-34; 5 mumol/L) also stimulated cAMP production, which was greater than that induced by 30 mumol/L KW-3902 alone. A significant increase in cAMP production by 30 mumol/L DPCPX was also observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of KW-3902, a novel adenosine A1 receptor antagonist, on sodium-dependent phosphate and glucose transport by the rat renal proximal tubular cell.

KW-3902, 1,3-dipropyl-8-(3-noradamantanyl)xanthine, is a novel potent and selective adenosine A1-receptor antagonist. KW-3902 has been found to cause significant diuresis and natriuresis. To investigate the action of this adenosine A1-receptor antagonist on phosphate transport in renal proximal tubular cells, we studied its effect on the uptake of phosphate by the cultured rat renal proximal tubular cell. KW-3902 significantly inhibited sodium-dependent uptake of phosphate at 10 minutes. The inhibitory effect was dose-dependent with maximum effect achieved at a KW-3902 concentration of 3 x 10(-5) M in the uptake media. The half-maximal inhibitory concentration, IC50, of KW-3902 on phosphate uptake was 2 x 10(-6) M. Dixon plot analysis of the uptake data was consistent with pure non-competitive inhibition. The inhibition constant, Ki, of 6.2 x 10(-6) M for phosphate transport, derived from the Dixon plot, was in close agreement with the IC50 calculated from a semilog dose response curve. Sodium-dependent glucose transport was not affected by KW-3902. These findings reveal that KW-3902 has a direct and specific inhibitory effect on phosphate uptake in renal proximal tubules.

Prostaglandin E2 and parathyroid hormone: comparisons of their actions on the rabbit proximal tubule.

Recent studies demonstrated that prostaglandin E2 (PGE2) participates in the regulation of glomerular and distal tubular function. A functional role for PGE2 on the proximal tubule has only recently been explored. Thus, we reported that PGE2 antagonizes the phosphaturic effect of PTH in the dog, which suggests that PGE2 may influence the transport functions of the mammalian proximal tubule. The present studies were designed to examine the direct effect of parathyroid hormone (PTH) and PGE2 on the fluxes of fluid (Jv) and phosphate (Jl-b PO4) in the rabbit proximal convoluted and straight tubules (PCT and PST). The activation of adenylate cyclase by the two agents was also examined. Finally, the combined effects of PTH and PGE2 on Jv and Jl-b PO4 were also studied in the PST. In the PCT, PTH (1 microgram/ml) inhibited Jv by 31% (P less than 0.05) but did not alter Jl-b PO4. PGE2 (10(-5) M) failed to act on either Jv or Jl-b PO4. In this segment, PTH stimulated adenylate cyclase but PGE2 did not. In the PST, PTH (1 microgram/ml) inhibited Jv and Jl-b PO4 by 34 and 20%, respectively (P less than 0.01). PGE2 (10(-5) M) also inhibited Jv and Jl-b PO4, by 33 and 12%, respectively (P less than 0.02). In contrast, PGF2 alpha, a related prostanoid, failed to influence these transport parameters. In the PST, PTH activated adenylate cyclase but PGE2 failed to do so. When both PTH (1 microgram/ml) and PGE2 (10(-7) M) were present simultaneously, Jv and Jl-b PO4 were comparable to that seen during control collections.(ABSTRACT TRUNCATED AT 250 WORDS)