Effects of the selective prostacyclin receptor antagonist RO3244019 on the micturition reflex in rats.

PURPOSE: We investigated the role of prostacyclin on afferent modulation of the micturition reflex using the novel selective prostacyclin receptor antagonist RO3244019 in rat models of bladder function. MATERIALS AND METHODS: The effects of RO3244019 on urodynamic parameters were evaluated in 3 rat models. In the anesthetized isovolumetric bladder contraction and the volume induced micturition reflex (Refill) models the effects of RO3244019 and chronic capsaicin desensitization were compared. In the citric acid induced detrusor overactivity model the effects of RO3244019 and the cyclooxygenase inhibitor indomethacin were evaluated. RESULTS: In the isovolumetric bladder contraction model RO3244019 dose dependently decreased bladder contraction frequency with a mean +/- SEM maximum decrease of 72.2% +/- 4.3% at 3.16 mg/kg. RO3244019 also dose dependently increased the micturition threshold in the Refill model with a maximum increase of 86.9% +/- 19.1% at 3.0 mg/kg. In animals that were chronically treated with capsaicin bladder contraction frequency was decreased by 88.9% in the isovolumetric bladder contraction model and micturition threshold was increased by 68.1% in the Refill model relative to sham treated rats. RO3244019 (3.0 mg/kg) further increased the micturition threshold in capsaicin treated animals by 53.7% +/- 18.1% from baseline. In the citric acid induced detrusor overactivity model citric acid decreased the voiding interval to 28.5% of baseline. This effect was reversed by RO3244019 (73.0% +/- 6.4%) and indomethacin (97.7% +/- 5.5%) at 3.0 mg/kg compared to vehicle (55.0% +/- 4.1%). CONCLUSIONS: The prostacyclin receptor antagonist RO3244019 decreased bladder contraction frequency and increased micturition threshold in the anesthetized isovolumetric bladder contraction and Refill models, respectively, and increased the micturition voiding interval in the conscious citric acid induced detrusor overactivity model. Additionally, RO3244019 remained effective for increasing the micturition threshold in the Refill model even following chronic capsaicin desensitization. Taken together these data suggest that prostacyclin may have a facilitory role in the micturition reflex by modulating the threshold for activation of capsaicin sensitive and insensitive bladder sensory afferents.

RO1138452 and RO3244794: characterization of structurally distinct, potent and selective IP (prostacyclin) receptor antagonists.

Prostacyclin (PGI2) possesses various physiological functions, including modulation of nociception, inflammation and cardiovascular activity. Elucidation of these functions has been hampered by the absence of selective IP receptor antagonists. Two structurally distinct series of IP receptor antagonists have been developed: 4,5-dihydro-1H-imidazol-2-yl)-[4-(4-isopropoxy-benzyl)-phenyl]-amine (RO1138452) and R-3-(4-fluoro-phenyl)-2-[5-(4-fluoro-phenyl)-benzofuran-2-ylmethoxycarbonylamino]-propionic acid (RO3244794).RO1138452 and RO3244794 display high affinity for IP receptors. In human platelets, the receptor affinities (pKi) were 9.3 +/- 0.1 and 7.7 +/- 0.03, respectively; in a recombinant IP receptor system, pKi values were 8.7 +/- 0.06 and 6.9 +/- 0.1, respectively. Functional antagonism of RO1138452 and RO3244794 was studied by measuring inhibition of carbaprostacyclin-induced cAMP accumulation in CHO-K1 cells stably expressing the human IP receptor. The antagonist affinities (pKi) of RO1138452 and RO3244794 were 9.0 +/- 0.06 and 8.5 +/- 0.11, respectively. Selectivity profiles for RO1138452 and RO3244794 were determined via a panel of receptor binding and enzyme assays. RO1138452 displayed affinity at I2 (8.3) and PAF (7.9) receptors, while RO3244794 was highly selective for the IP receptor: pKi values for EP1 (< 5), EP3 (5.38), EP4 (5.74) and TP (5.09). RO1138452 (1-10 mg kg(-1), i.v.) and RO3244794 (1-30 mg kg(-1), i.v.) significantly reduced acetic acid-induced abdominal constrictions. RO1138452 (3-100 mg kg(-1), p.o.) and RO3244794 (0.3-30 mg kg(-1), p.o.) significantly reduced carrageenan-induced mechanical hyperalgesia and edema formation. RO3244794 (1 and 10 mg kg(-1), p.o.) also significantly reduced chronic joint discomfort induced by monoiodoacetate. These data suggest that RO1138452 and RO3244794 are potent and selective antagonists for both human and rat IP receptors and that they possess analgesic and anti-inflammatory potential.

The role of the prostacyclin receptor antagonist RO3244019 in treating neurogenic detrusor overactivity after spinal cord injury in rats.

OBJECTIVE: To determine the effects of the prostacyclin receptor (IP) antagonist RO3244019 on neurogenic detrusor overactivity (NDO) in spinal cord-injured (SCI) neurogenic bladder of the rat. MATERIALS AND METHODS: Female Sprague-Dawley rats with SCI were divided into four treatment groups of eight each: vehicle (200 mm Tris base), indomethacin (3 mg/kg), RO3244019 (at 1 and 5 mg/kg). The conscious rats were assessed by cystometry, by slowly infusing the bladder with physiological normal saline at 0.04 mL/min. After 1 h of cystometry one of the four compounds was administered intravenously to the rats and changes in cystometrogram tracings recorded. Seven voiding variables were calculated before and after administering each compound: the intercontractile interval (ICI) for all contractions, voiding ICI, amplitude of all contractions, amplitude of voiding contractions, time to first void (TFV), voided volumes (VVs), and first VV. Data were analysed using a paired t-test for each of the experiments. RESULTS: At 1 mg/kg, the RO compound was associated with a statistically significant difference in the voiding ICI and VVs (both P < 0.05). The mean (sd) voiding ICI increased from 621 (140) to 889 (119) s (43% increase) and the VVs from 0.53 (0.13) to 0.72 (0.09) mL (36% increase). However, there was no statistically significant difference in the TFV or the first VVs. Increasing the dose to 5 mg/kg was more effective in improving the voiding ICI and the VVs (both P < 0.01). The voiding ICI increased from 716 (130) to 1346 (159) s (88% increase) and the VVs from 0.60 (0.11) to 1.05 (0.12) mL (75% increase). In addition, the higher dose had a statistically significant difference in the TFV (P < 0.05). There was more than a four-fold increase in the TFV, from 807 (138) to 3239 (883) s. At 5 mg/kg, the difference in the first VV before and after administering the compound was also almost statistically significant (P = 0.057); the first VV increased from 0.56 (0.14) to 1.01 (0.21) mL. There were no statistically significant differences in the amplitude of contractions or the ICI for all contractions for either of the dosages. Indomethacin at 3 mg/kg was the most effective compound for improving all of the voiding variables and was the only one to show a significant difference in the first VV. However, the IP antagonist at 5 mg/kg was almost as effective as indomethacin when comparing other variables, e.g. the voiding ICI and the VV. There was no statistically significant difference in any of the seven voiding variables before and after administering the vehicle. CONCLUSION: The IP antagonist RO3244019 was effective in treating NDO in SCI bladders. While RO3244019 at 1 mg/kg significantly increased the voiding ICI and VVs, 5 mg/kg appeared to be more effective, suggesting a dose-dependent effect of the drug. The RO compound at 5 mg/kg was almost as effective as indomethacin in improving all of the voiding variables.

Discovery and SAR development of 2-(phenylamino) imidazolines as prostacyclin receptor antagonists [corrected].

On the basis of screening hits (1a,b), a series of selective, high affinity prostacyclin receptor antagonists was developed. The optimized lead compound 25d [(4,5-dihydro-1H-imidazol-2-yl)-[4-(4-isopropoxybenzyl)phenyl]amine] had analgesic activity in the rat.