Leukotriene receptor antagonists. III. Pharmacological and chemical studies with LY137617, a leukotriene D4 and leukotriene E4 receptor antagonist.

A series of chlorophenoxyalkyl acids were prepared and evaluated as pharmacological antagonists of leukotriene D4. Structure-activity relationship studies pointed to LY137617 as a compound with possible therapeutic value. In experiments on isolated smooth muscles from the guinea-pig, this agent was a selective and moderately potent antagonist of leukotriene D4 and also leukotriene E4. Other in vitro experiments demonstrated that LY137617 had a high affinity for protein molecules. This was reflected in vivo as a weaker than expected efficacy against leukotriene-mediated events, limiting the compound's potential as a clinical candidate. Nevertheless, agents of this type will prove useful in the laboratory to increase knowledge of leukotriene receptor-antagonist interactions.

Leukotriene receptor antagonists. 2. The [[(tetrazol-5-ylaryl)oxy]methyl]acetophenone derivatives.

A series of [[(tetrazol-5-ylaryl)oxy]methyl]acetophenones was synthesized and evaluated as antagonists of leukotriene D4 induced contractions of guinea pig ileum. Substitutions at the 3-position of the acetophenone with ethyl (66), propyl (68), butyl (83), and isobutyl (84) gave -log IC50 values of 7.9, 8.0, 7.8, and 7.7, respectively. Equally potent compounds were obtained when the tetrazol-5-yl group was connected to the second benzene ring in the para position with a chemical bond (67), methylene (68), or ethylene (71). For retention of high antagonist activity, the acetophenone should be substituted in the 2-position by a hydroxyl group and the tetrazole ring should have an acidic hydrogen atom. 1-[2-Hydroxy-3-propyl-4-[[4-(1H-tetrazol-5-ylmethy) phenoxy]methyl]phenyl]ethanone (68, LY1632443) has undergone extensive pharmacologic evaluation for its potential as an antiasthma agent.

Leukotriene receptor antagonists. 1. Synthesis and structure-activity relationships of alkoxyacetophenone derivatives.

A series of derivatives of 2,4-dihydroxy-3-propylacetophenone(1) were prepared and examined for their ability to block leukotriene D4 (LTD4) induced contraction of guinea pig ileum. Straight-chain carboxylic acids where the carboxyl group was separated from the acetophenone moiety by varying numbers of methylenes were evaluated, and maximum activity was obtained with the pentamethylene acid (6). Examination of ring substitution showed that the 2-propyl-3-hydroxy-4-acetyl substitution pattern was required for maximum LTD4 antagonist activity. Additional chain terminal groups were examined, and the acidic 5-tetrazolyl group separated from the acetophenone moiety by four to seven methylenes (26, 23, 27, 28) gave excellent in vitro and in vivo activities. Compound 26 (LY171883) had the best balance of in vitro and in vivo activity. It lacked bronchospastic activity at the doses administered and has been chosen for clinical evaluation.

Pharmacologic analysis of LY188695 (KB-2413), 1-(2-ethoxyethyl)-2-(4-methyl-1-homopiperazinyl)-benzimidazole difumarate, a potent histamine1 receptor antagonist.

LY188695 was evaluated both in vitro and in vivo in the guinea pig to determine its pharmacologic profile. The compound antagonized histamine-induced contractions of ileum, aorta, and trachea with pKB values of 9.9, 9.9, and 9.2 respectively. In the lung parenchymal strip, LY188695 caused a rightward shift of the histamine concentration-response curve with a reduction in the maximal response at all antagonist concentrations tested. The reason for this effect is unknown, but it was not due to a nonspecific depressant action of the compound on the parenchyma. Selectivity was shown by its inactivity against leukotriene D4, bradykinin, prostaglandin F2 alpha, acetylcholine, norepinephrine, and serotonin on various guinea pig and rat smooth muscles. Similarly, H2 receptor-mediated relaxation of the rat uterus was unaltered by LY188695. Increases in total pulmonary impedance caused by i.v. histamine to anesthetized guinea pigs were reduced by as little as 3 micrograms/kg given orally 1 hour prior to histamine challenge. In this system, LY188695 was 15 times more potent than chlorpheniramine and 100 times more potent than terfenadine. Similar responses elicited by acetylcholine were not antagonized by LY188695. A duration of action greater than 4 hours was observed in this model. Ovalbumin given i.v. to sensitized guinea pigs increased total pulmonary impedance which was markedly decreased after oral administration of 30 or 100 micrograms/kg LY188695. These results indicate that LY188695 is a very potent antagonist of H1-mediated responses and suggest that this agent might be useful in disease states characterized by an overproduction of histamine.

Evaluation of LY163443, 1-[2-hydroxy-3-propyl-4-([4- (1H-tetrazol-5-ylmethyl)phenoxy]methyl) phenyl]ethanone, as a pharmacologic antagonist of leukotrienes D4 and E4.

LY163443,1-[2-hydroxy-3-propyl-4-([4- (1H-tetrazol-5-ylmethyl)phenoxy]- phenoxy]methyl)phenyl]ethanone, antagonized LTD4-induced contractions of guinea pig ileum, trachea, and lung parenchyma. Tracheal contractions to LTE4 were also inhibited by LY163443. The compound had minimal effect against ileal responses to LTC4 and parenchymal contractions to LTB4. Furthermore, LY163443 had little to no effect against contractions of isolated smooth muscles to histamine, bradykinin, PGF2 alpha, carbachol, serotonin or U46619. LY163443, given by oral administration to guinea pigs, blocked LTD4-induced increases in total pulmonary impedance (TPI). Similar responses elicited by histamine or U46619 were unaffected. Increases in TPI in response to i.v. administration of LTC4 were antagonized by LY163443 given by the same route. Ovalbumin challenge also increased TPI in guinea pigs previously sensitized against this antigen. In such animals, pretreated with pyrilamine, propranolol, and indomethacin, oral administration of LY163443 blocked the increase in TPI caused by ovalbumin. Additionally, LTD4 given intradermally to guinea pigs caused a vascular leakage which was suppressed by prior oral administration of LY163443. Finally, LY163443 relaxed isolated guinea pig trachea previously contracted with LTD4, histamine, or carbachol. Relaxation of tissues contracted by these latter two agonists suggested some inherent airway smooth muscle relaxant properties of the molecule. This was further demonstrated by showing some bronchodilator activity in an in vivo setting. Thus, this pharmacologic profile indicates that LY163443, or a member of the same chemical family, warrants consideration as a possible therapeutic agent in the treatment of asthma and in diseases characterized by an overproduction of LTD4 and LTE4.

LY171883, 1-less than 2-hydroxy-3-propyl-4-less than 4-(1H-tetrazol-5-yl) butoxy greater than phenyl greater than ethanone, an orally active leukotriene D4 antagonist.

LY171883, 1-less than2-hydroxy-3-propyl-4-less than 4-(h-tetrazol-5-yl)butoxy greater than phenyl greater than ethanone, proved to be a potent antagonist of leukotriene (LT) D4 in guinea-pig ileum, trachea and lung parenchyma. The compound had little or no effect on contractions of isolated tissues to LTB4, prostaglandin F2 alpha, serotonin, histamine, bradykinin or carbamycholine. Responses of trachea to U46619, a thromboxane A2 mimetic, were antagonized by LY171883, but the doses required were approximately 10-fold higher than those necessary to produce the same degree of antagonism against LTD4. U46619 produced weak ileal contractions that were not blocked by LY171883. LY171883 antagonized both LTD4- and antigen-induced increases in total pulmonary resistance in anesthetized guinea pigs. LTD4 given intradermally to guinea pigs caused vascular leakage which was suppressed by prior administration of LY171883. LTC4-induced contractions of isolated ilea were only minimally antagonized by LY171883 whereas this agent reduced LTC4-evoked increases in total pulmonary resistance. Trachea contracted by LTD4 were relaxed by LY171883. Likewise, trachea contracted by either histamine or carbamylcholine were relaxed by LY171883 suggesting that this compound has airway smooth muscle relaxing properties. In vivo experiments supported these observations. In concert with these findings, biochemical studies showed LY171883 to be a potent inhibitor of phosphodiesterase obtained from human polymorphonuclear leukocytes and various guinea-pig tissues. This pharmacologic analysis indicates that LY171883, or a congener, may be of therapeutic value in asthma and in disease states characterized by an overproduction of LTD4.

Pharmacologic analysis of two novel inhibitors of leukotriene (slow reacting substance) release.

LY83583 , a quinolinedione , and LY151364 , a quinoxalinedione , were developed as inhibitors of leukotriene (slow reacting substance of anaphylaxis) release. They preferentially inhibited the release of leukotrienes over histamine from fragmented guinea-pig lung and rat peritoneal cells in vitro, regardless of whether the mediators were released immunologically by antigen or chemically by the divalent cationic ionophore, A23187. Similar results were obtained with rat peritoneal cells in vivo. In that system, comparison of LY83583 with disodium cromoglycate showed the former to preferentially inhibit release of leukotrienes, whereas the latter favored inhibition of histamine release. LY83583 did not significantly decrease antigen-induced bronchospasm in guinea pigs after i.v. administration of doses that approached toxic levels. In addition, LY83583 did not antagonize contractions to carbachol or histamine on guinea-pig trachea, prostaglandin F2 alpha-elicited contraction on guinea-pig ileum or contractions produced by serotonin on guinea-pig aorta. This agent, at 1 X 10(-5) M, reduced the maximal responses to bradykinin on ileum and caused a rightward displacement with a reduction in the maximal response to norepinephrine on guinea-pig aorta. In summary, LY83583 and LY151364 have interesting pharmacologic profiles which make them useful as tools in understanding the role of the leukotrienes in isolated tissue systems.

Slow reacting substance of anaphylaxis (SRS-A) release from guinea-pig lung parenchyma during antigen- or ionophore-induced contraction.

A dual isolated organ technique comprised of a guinea-pig lung parenchymal strip and a guinea-pig ileum was used to determine if slow reacting substance of anaphylaxis (SRS-A) is released from parenchyma during contractions evoked by antigen (ovalbumin) or by ionophore (A23187). An immunologically sensitized parenchyma served as the primary target organ for ovalbumin and either a sensitized or unsensitized parenchyma was the target tissue for A23187; an unsensitized ileum functioned as the assay organ. In the presence of pyrilamine and indomethacin, ovalbumin or A23187 produced contractions of the parenchyma and concomitantly caused release of SRS-A from the lung strip which was indicated by a contraction of the ileum. The ileal response was antagonized by FPL 55712, whereas the parenchyma contractions were unaffected. Additional experiments were conducted in which parenchyma was contracted with histamine. At the height of the histamine contraction, the bathing fluid surrounding the parenchyma was removed and assayed on a pyrilamine-treated ileum. SRS-A was not detected, indicating that SRS-A release from parenchyma is not a function of tissue contraction per se, but is related to the antigen- and ionophore-induced contractions. To explain the lack of effect of FPL 55712 on parenchymal contractions to antigen or ionophore, we compared the degree of antagonism produced by FPL 55712 on SRS-A contraction of parenchyma and ileum. These experiments indicated the possibility that at least two different classes of SRS-A receptors exist and that those in the ileum and lung differ.

Schultz-Dale reaction in mouse trachea.

A method was developed to induce contraction of immunologically sensitized mouse trachea by antigen (Schultz-Dale reaction). The response was mediated by immunoglobulin (Ig) E antibody directed against either the hapten DNP, the hapten carrier conjugate DNP-keyhole limpet hemocyanin (KLH), or the unmodified carrier KLH. Tracheal contractions were elicited by DNP-KLH, KLH, or DNP-bovine serum albumin (BSA) but not by DNP or BSA alone. This procedure represents a useful index of in vitro anaphylaxis in mouse airway smooth muscle.