Kiyoto Sakata

The contractile action of leukotriene B4 in the guinea-pig lung involves a vascular component.

Leukotriene B4 (LTB4) is a potent leukocyte chemoattractant, acting on specific receptors, BLT receptors. The aim of this study was to examine the mechanism of action of LTB4 in the guinea‐pig lung, using strips of lung parenchyma (GPLP), spirals of trachea (GPT) and bronchus (GPB) and rings of pulmonary artery (GPPA). Mechanical responses were studied in organ baths, and mediator release was assessed using enzyme immuno assay. LTB4 induced similar contractions of GPLP and GPPA, whereas LTB4 had only small contractile effects in GPT and GPB. In addition, the contractile response to LTB4 was reproduced in the human pulmonary artery. In the GPLP, the unselective BLT receptor antagonist ONO‐4057 abolished the contractions induced by LTB4, whereas the selective BLT1 receptor antagonist U–75302 only partly inhibited the LTB4‐induced contractions. In the GPPA, both antagonists abolished the response to LTB4. The effect of LTB4 in GPPA and GPLP was indirect and mediated by the release of thromboxane A2 and histamine, as supported by selective pharmacologic interventions and measurements of thromboxane B2 and histamine in the organ baths. In conclusion, the results indicate a new biological function of LTB4, namely to constrict isolated pulmonary arteries. Moreover, the findings suggest that the LTB4‐induced contractions of GPPA were mediated by a BLT1 receptor, whereas BLT2 receptor activation accounted for a major part of the contraction of GPLP, making the latter preparation a suitable assay for BLT2 receptors.

Design and synthesis of a selective EP4-receptor agonist. Part 4: practical synthesis and biological evaluation of a novel highly selective EP4-receptor agonist

A practical method of synthesizing a highly selective EP4-receptor agonist 1 using Corey lactone 2 as a key intermediate was developed. Selective methanesulfonylation of the primary alcohol of the diol 8 under the newly devised conditions followed by the protection of the remaining secondary alcohol are key reactions in this new method. Further biological evaluation of 1a-b is also reported.

Design and synthesis of a highly selective EP4-receptor agonist. Part 2: 5-thia and 9β-haloPG derivatives with improved stability

Further chemical modification to identify more chemically stabilized EP4-receptor selective agonists was continued. As a result, a further two EP4-receptor selective agonists 5-thiaPGE(1) 2a, 10 and 9beta-chloroPGF(2) analogue 11 were discovered.

Receptor preferences of cysteinyl-leukotrienes in the guinea pig lung parenchyma

Two cysteinyl-leukotriene receptors, CysLT(1) and CysLT(2) receptors, have been cloned, but the contractions to cysteinyl-leukotrienes in the guinea pig lung parenchyma have been reported to be resistant to CysLT(2) receptor antagonism and to be only partially inhibited by CysLT(1) receptor antagonism. The receptor preferences of the individual cysteinyl-leukotrienes (leukotriene C(4), D(4) and E(4)) in the guinea pig lung parenchyma were studied in organ baths. CysLT(1) receptor antagonists competitively inhibited the contraction to leukotriene E(4), but exhibited only weak antagonism of contractions to leukotriene C(4) and D(4). In the presence of the cyclooxygenese inhibitor indomethacin and the nitric oxide synthase inhibitor N(omega)-nitro-L-arginine (L-NOARG), the CysLT(1) receptor antagonists did not further inhibit the leukotriene D(4)-induced contraction. These results suggest that leukotriene E(4) solely activates a CysLT(1) receptor, and that the CysLT(1) receptor antagonist-resistant contraction to leukotriene D(4) and C(4) is mediated via another CysLT receptor.

Synthesis and evaluation of novel modified γ-lactam prostanoids as EP4 subtype-selective agonists

To identify chemically and metabolically stable subtype-selective EP4 agonists, design and synthesis of a series of modified γ-lactam prostanoids has been continued. Prostanoids bearing 2-oxo-1,3-oxazolidine, 2-oxo-1,3-thiazolidine and 5-thioxopyrrolidine as a surrogate for the γ-hydroxycyclopentanone without a troublesome 11-hydroxy group were identified as highly subtype-selective EP4 agonists. Among the tested, several representative compounds demonstrated in vivo efficacy after oral dosing in rats. Their pharmacokinetic and structure-activity relationship studies are presented.