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Prostaglandins (PGs) in the E-series exhibit potent gastric antisecretory activity, but can also cause diarrhea, which is mediated via PGE receptors. SC-46275, an omega-chain cyclopentenyl analog of the E-type PG enisoprost, was evaluated with other E-PGs for PGE receptor binding activity in gastric and intestinal tissues. SC-46275, enisoprost, misoprostol and PGE1 were first evaluated in enriched canine gastric parietal cells with [3H]misoprostol free acid binding and subsequently with [3H]PGE1 binding in canine intestinal tissues where misoprostol free acid had weak receptor binding activity. The receptor binding potency of SC-46275 (IC50, 0.013 mM) in enriched canine parietal cell preparations was found to be much greater than misoprostol and enisoprost (IC50, 10 and 8 nM), whereas PGE1 had the least potency (IC50, 37 nM). Similar relative potencies for these PGs were also obtained in the inhibition of histamine-stimulated acid secretion in enriched parietal cell preparations. In small intestinal mucosal and muscle membranes, the receptor binding potency of SC-46275 (IC50, 13 and 20 microM) was much less than misoprostol or enisoprost (IC50, 0.39-1.2 microM) and substantially less than PGE1 (IC50, 0.017 and 0.066 microM). This weak binding activity of SC-46275 in intestinal tissues is consistent with its reported weak diarrheagenic activity in the rat. These results suggest that SC-46275 binds preferentially to gastric vs. intestinal PGE receptors and is specific for the EP3 receptors.

Prostaglandin E (PGE) receptors in canine small intestinal mucosal and muscle membrane preparations were labeled with [3H] PGE1. Saturable, high affinity binding of [3H] PGE1 was observed in both preparations. The density of binding sites (fmol/mg protein) was 39 for mucosal membranes and 60 for muscle membranes, with corresponding dissociation constants of 10.6 nM and 5.8 nM, respectively. [3H] PGE1 binding sites in both preparations showed stereospecificity and high affinity for natural PGE1 and PGE2, but not for I or F-type PGs. Synthetic PGEs such as misoprostol and enisoprost had lower affinity than PGE1 or PGE2. Several analogs of enisoprost bound weakly to the binding sites. A highly significant correlation (C.C. = 0.9) was demonstrated between mucosal and muscle binding potency for a series of enisoprost analogs. There was also a significant positive correlation between the receptor binding potency and rat diarrheagenic activity for these analogs. These results indicate that PGE receptors in canine intestinal mucosa and muscle can be directly studied with [3H] PGE1 binding. The mucosal and muscle PGE receptors may have similar ligand binding specificity. We speculate that these receptors are likely to be associated with the diarrheagenic activity of PGEs.

We have examined the effect of caerulein on intestinal fluid transport in vivo. Rat jejunal, ileal, and colonic segments were perfused with a physiologic buffer for a 60-min control period, followed by a 60-min period after caerulein, prostaglandin E2 (PGE2), or saline (0.9%, w/v) were given by intramuscular (i.m) injection. At a dose of 5 micrograms/kg caerulein had no effect on ileal fluid transport. Lower doses of caerulein (300 ng and 1 microgram/kg) also had no effect. PGE2 significantly (P less than 0.05) inhibited net ileal absorption by 94 +/- 24% from pre-dose levels. Caerulein (300 ng/kg) had no effect (P greater than 0.05) on jejunal or colonic fluid transport. Intestinal fluid accumulation assessed by the 'enteropooling' assay did not increase after 30 ng or 1 microgram/kg of caerulein, whereas in animals given PGE2 of (5 mg/kg) the fluid accumulation more than doubled (P less than 0.001): control 1.59 +/- 0.15 ml; cerulein (1 microgram/kg) 1.36 +/- 0.20 ml; and PGE2 4.7 +/- 0.50 ml. Serum levels of caerulein (after a 1 microgram/kg dose), measured by radioimmunoassay, were elevated up to 30 min after i.m. injection. The data indicate that caerulein has no direct effect on rat small or large intestinal fluid transport.