Leukotriene Synthesis Inhibitor Research Articles

Protection of the myocardium during myocardial infarction: Pharmacologic protection during thrombolytic therapy

The most effective way to protect injured myocardium during the early phases of evolving myocardial infarction is with reperfusion. However, reperfusion may induce further myocardial injury. Alterations in the major determinants of myocardial oxygen demand, including increases in heart rate, contractile state and myocardial wall tension may also increase infarct size. An understanding of the basic mechanisms involved in the evolution of myocardial injury during myocardial infarction is important to the development of protective strategies that may reduce infarct size or attenuate reperfusion-induced myocardial injury, or both. Selective calcium antagonists, β blockers, and phospholipase antagonists or inhibitors may reduce infarct size in association with early reperfusion. Reperfusion injury may be attenuated by free radical scavengers and calcium antagonists. Reinfarction, after the initial event, may ultimately be reduced in frequency by pharmacologic strategies that interfere with platelet adhesion, aggregation, and mediator release from activated platelets and white cells that promote further platelet aggregation or induce coronary artery vasoconstriction. Included among protective strategies are thromboxane receptor antagonists and synthesis inhibitors, serotonin receptor antagonists, possibly leukotriene synthesis inhibitors or receptor antagonists, and aspirin. Future clinical trials should test the combined efficacy of reperfusion and selected pharmacologic strategies that after calcium accumulation in the injured myocardium, diminish the injurious effects of β- and α-adrenergic mechanisms and oxygen-derived free radicals on injured myocytes, and prevent reocclusion mediated by platelets and platelet mediators.

Treatment of Ulcerative Colitis with Fish Oil n-3-ω-Fatty Acid

We evaluated the efficacy of fish oil n--3-omega-fatty acids, inhibitors of leukotriene synthesis, in the treatment of ulcerative colitis. An open trial of 10 patients with mild to moderate ulcerative colitis who had either failed (n = 9) or refused (n = 1) conventional therapy was performed. Patients received 15 MAX-EPA capsules containing a total of 2.7 g of eicosapentanoic acid in three divided doses daily for 8 weeks. The activity of ulcerative colitis and response to therapy was based upon daily stool diaries, sigmoidoscopy, and symptomatic response. All patients tolerated the fish oil and showed no alteration in routine blood studies. Seven patients had moderate to marked improvement; steroid dose could be reduced in four of the five patients on prednisone. Three patients had little or no improvement. No patient worsened. These results of our open study appear to justify a double-blind trial of this dietary supplement in ambulatory patients with ulcerative colitis.

An orally active inhibitor of leukotriene synthesis accelerates healing in a rat model of colitis

Leukotrienes (LTs) have been implicated as mediators of the inflammation and ulceration associated with ulcerative colitis and Crohn's disease. In the present study, the effects of a novel, orally active inhibitor of LT synthesis (MK-886) were examined in a rat model of chronic colitis. Colitis was induced by intracolonic administration of trinitrobenzenesulfonic acid. Colonic LTB4 synthesis was measured after incubation of tissue samples in vitro and by in vivo equilibrium dialysis. A single dose of MK-886 (10 mg/kg) significantly inhibited colonic LTB4 synthesis for greater than 24 h. Daily treatment with this dose significantly reduced colonic damage, as assessed macroscopically and histologically, when the treatment was performed 2 h before induction of colitis and daily thereafter for 1 wk, but not when treatment was performed during the second week after induction of colitis. A less marked beneficial effect of MK-886 was observed when the pretreatment dose was excluded, suggesting a role for LTs in the early events of the inflammatory process. Inhibition of LT synthesis during the first 24 h after induction of colitis did not alter the extent of infiltration of neutrophils into the colon, as measured by tissue myeloperoxidase activity. Daily treatment with sulfasalazine (100 mg/kg po) either during the first or second week after induction of colitis did not significantly affect the rates of healing. At the dose used, sulfasalazine only produced a transient inhibition of colonic LTB4 synthesis. This study therefore demonstrates that a specific, orally active inhibitor of LT synthesis can significantly accelerate healing in this animal model of colitis when the treatment is performed during the early phase of the inflammatory response.

The effect of leukotriene synthesis inhibitors on hamster periodontitis

The purpose of these experiments was to assess the role of the lipoxygenation products of arachidonic acid in hamster periodontitis. Phenidone and ketoconazole were used as inhibitors of leukotriene synthesis. In an established periodontitis, both drugs administered for 30 days induced a statistically significant decrease in PMNLs in the infiltrated connective tissue and around bacterial plaque within periodontal pockets. These changes were associated with a significant decrease in osteoclastic bone resorption. The results suggest that leukotrienes, and particularly leukotriene B 4, are involved during hamster periodontitis and are responsible for PMNL infiltration of the periodontal pocket. The effects on bone are probably the consequence of the reduced inflammation resulting from the decrease in PMNL chemotaxis.

MK886, a potent and specific leukotriene biosynthesis inhibitor blocks and reverses the membrane association of 5-lipoxygenase in ionophore-challenged leukocytes.

Recently, we have shown that ionophore activation of human leukocytes results in leukotriene synthesis and a translocation of 5-lipoxygenase from the cytosol to cellular membrane. This membrane translocation was postulated to be an important early activation step for the enzyme. 3-[1-(p-Chlorobenzyl)-5-(isopropyl)-3-tert-butylthioindol-2-yl]-2, 2- dimethylpropanoic acid (MK886) is a potent and specific inhibitor of leukotriene biosynthesis in vivo and in intact cells, but has no direct effect on 5-lipoxygenase activity in cell-free systems. In this report, we show that MK886 can both prevent and reverse the membrane translocation of 5-lipoxygenase, in conjunction with the inhibition of leukotriene synthesis. Similar compounds of the indole class could also inhibit the membrane translocation of 5-lipoxygenase in a rank order of potency that correlated with their potencies for leukotriene synthesis inhibition. In contrast L-656,224, a direct 5-lipoxygenase inhibitor, had no effect on the translocation of the enzyme. Attempts to demonstrate the effects of MK886 on the association of 5-lipoxygenase with membrane in cell-free preparations failed due to a nonspecific Ca2+-dependent sedimentation of the enzyme. The mechanism of action of MK-886 is therefore to block translocation, prevent subsequent activation of 5-lipoxygenase, and hence block cellular leukotriene biosynthesis.

Piriprost Pretreatment Attenuates the Smoke-Induced Increase in99mTcDTPA Lung Clearance

We studied the effects of acute smoke exposure on lung permeability, eicosanoids, and inflammatory cell activity. Thirty-five New Zealand white rabbits were anesthetized, paralyzed, and exposed to 60 tidal volume breaths of diesel fuel-polycarbonate plastic smoke or sham smoke within 10 min. At 1 h postexposure the rabbits were killed and their lungs were removed for bronchoalveolar lavage (BAL) or pathologic procedures. Smoke exposure caused decreases in technetium-labeled diethylenetriamine pentaacetate (99mTcDTPA, mol. wt. 492 Da) biological half-life (t1/2), BAL plasminogen activator, and BAL leukotriene B4 (LTB4). In addition, alveolar macrophage acid phosphatase enzyme activity increased in smoke-exposed rabbits. The leukotriene synthesis inhibitor, piriprost (U-60,257), given before smoke exposure, caused attenuation of the changes in 99mTcDTPA uptake and plasminogen activator, swelling of type I alveolar cell epithelium, a large increase in lung inflammatory cells, and decreases in BAL LTB4, prostaglandin E2 (PGE2), and TxB2 (stable metabolite of thromboxane, TxA2). We conclude that changes in alveolar-capillary barrier permeability and plasminogen activator activity occur within 1 h after exposure to smoke and may play an early role in the inflammatory process associated with smoke inhalation injury. Furthermore, piriprost attenuates the smoke-induced increase in alveolar-capillary barrier permeability and decrease in plasminogen activator activity and causes a swelling of type I alveolar epithelium. However, our data suggest that neither lung eicosanoids or the alveolar macrophage lysis process plays a major role in the smoke-induced increase in alveolar-capillary barrier permeability.

Cold Injury, Blood–Brain Barrier Changes, and Leukotriene Synthesis: Inhibition by Phenidone

Transcranial cold injury in rats and guinea pigs induced cerebral extravasation of albumin labeled with Evans blue dye or 125I, respective indicators of the area and amount of blood-brain barrier (BBB) disruption. Radioimmunoassay of brain extracts showed that cold injury induced leukotriene (LT)C4 in rat and guinea pig brains 15 min after injury. In guinea pigs, the LT synthesis inhibitor phenidone (30 mg/kg, i.p.) completely blocked cold-induced LTC4 in brain. Phenidone (30 and 100 mg/kg) also inhibited cerebral tissue accumulation of 125I-albumin and dye in rats and guinea pigs. Phenidone is reported to show antioxidant properties and selective lipoxygenase inhibition of arachidonic acid metabolism compared to cyclooxygenase inhibitors, meclofenamate sodium, and other nonsteroidal anti-inflammatory agents. Since several oxygen and hydroxyl radical scavengers and the cyclooxygenase inhibitor, meclofenamate sodium, did not inhibit protein extravasation, the findings support a role for LT as a mediator of cold-induced changes in BBB permeability in rats and guinea pigs and suggest that the inhibitory effects of phenidone on BBB permeability may be due to inhibition of LT production.

The synthesis, release and action of leukotrienes in the isolated, unstimulated, buffer-perfused rat heart

When the perfusion medium of an isolated, non-recirculating, Langendorff rat heart is changed from Krebs buffer to coronary effluent, a significant vasoconstriction (23%, P < 0.005) is observed. In this study we have investigated the involvement of leukotrienes in this phenomenon. We have extracted and quantified leukotrienes C 4, D 4 and E 4 in samples of coronary effluent taken at different times during the first 2 h of perfusion; the otal amounts released during this time were 9, 5 and 32 pmol of LTC 4, LTD 4 and LTE 4 respectively. We have used two different methods to prevent the action of the effluent leukotrienes on the heart. Firstly, we have blocked the leukotriene receptors in the heart, with FPL 55712 (3.8 μ m), during perfusion with effluent and, secondly, we have perfused with coronary effluent which was collected in the presence of a leukotriene synthesis inhibitor, AA861 (1 μ m). The addition of FPL 55712 to the effluent decreased the normally observed vasoconstriction such that after 30 min the coronary flow rate (CFR) was 114 ± 3% ( n = 6) compared with 66 ± 1% ( n = 7) with effluent alone ( P < 0.005). Effluent collected in the presence of AA861 also caused a decrease in the normally observed vasoconstriction such that by 30 min the CFR was still 88 ± 2% ( n = 6, P < 0.005 compared to controls). We have confirmed the proposed involvement of leukotrienes in the effluent-induced vasoconstriction by investigating the effect of a mixture of the synthetic leukotrienes C 4, D 4 and E 4, when each of them was present at the same concentration as measured in the coronary effluent; the vasoconstriction observed was superimposable upon that seen with effluent. This vasoactive effect of the leukotriene mixture was not secondary to a change in contractility, since this only decreased to 97 ± 5% ( n = 9) during the 30 min of the leukotriene infusion. Finally, we have studied the effects of the same two leukotriene blockers in normal, buffer-perfused hearts after an initial perfusion of either 30 or 120 min. Application of either AA861 or FPL 55712 resulted in a dramatic vasodilatation (25 to 45% increase), a larger effect always being observed after the shorter initial period of perfusion. Our conclusions are two-fold. Firstly, isolated, buffer-perfused rat hearts synthesize leukotrienes C 4, D 4 and E 4 in considerable amounts and release them into the coronary effluent and secondly, the coronary flow rates of isolated, buffer-perfused rat hearts are partly controlled by the action of internally produced leukotrienes.

1-[4-[3-[4-[Bis (4-fluorophenyl) hydroxymethyl]-1-piperidinyl] propoxy]-3]-methoxyphenyl] ethanone (AHR-5333): A selective human blood neutrophil 5-lipoxygenase inhibitor

In this study we report the in vitro inhibition of leukotriene synthesis in calcium ionophore (A23187)-stimulated, intact human blood neutrophils by AHR-5333. The results showed that AHR-5333 inhibits 5-HETE, LTB 4 and LTC 4 synthesis with IC 50 values of 13.9, 13.7 and 6.9 μM, respectively. Further examination of the effect of AHR-5333 on individual reaction of the 5-lipoxygenase pathway (i.e. conversion of LTA 4, to LTB 4, LTA 4 to LTC 4, and arachidonic acid to 5-HETE) showed that this agent was not inhibitory to LTA 4 expoxyhydrolase and glutathione-S-transferase activity in neutrophil homogenates. However, conversion of arachidonic acid (30 μM) to 5-HETE was half maximally inhibited by 20 μM AHR-5333 in the cell-free system. The inhibition of LTB 4 and LTC 4 formation in intact neutrophils by AHR-5333 appears to be entirely due to a selective inhibition of 5-lipoxygenase activity and an impaired formation of LTA 4, which serves as substrate for LTA 4 epoxyhydrolase and glutathione-S-transferase. AHR-5333 did not affect the transformationof exogenous arachidonic acid to thromboxane B 2, HHT and 12-HETE in preparations of washed human platelets, indicating that this agent has no effect on platelet prostaglandin H synthase, thromboxane synthase and 12-lipoxygenase activity. The lack of inhibitory activity of AHR-5333 on prostaglandin H synthase activity was confirmed with microsomal preparation of sheep vesicular glands.

Orally administered tolfenamic acid inhibits leukotriene synthesis in isolated human peripheral polymorphonuclear leukocytes

Special interest has been focused on the development of dual inhibitors of the cyclo-oxygenase and lipoxygenase pathways of arachidonic acid metabolism. In contrast to other classic NSAIDs, some fenamates in clinically achievable concentrations have been shown to inhibit synthesis of 5-lipoxygenase products in vitro. In the present work, we studied the effect of orally administered tolfenamic acid (600 mg) on Ca ionophore A 23187 -induced leukotriene synthesis in isolated human polymorphonuclear leukocytes. Leukotriene production was reduced in all 14 subjects studied, the mean inhibition of LTB4 synthesis being 16 +/- 3% and that of LTC4 33 +/ 7%. The inhibition correlated positively with serum acid concentrations. We suggest that inhibition of leukotriene synthesis is an additional mechanism of the anti-inflammatory, antimigraine and antidysmenorrhoeic effects of tolfenamic acid, and a possible explanation for its rare gastric and bronchoconstrictive side-effects.

Wy-48,252 (1,1,1-trifluoro-N-[3-(2-quinolinylmethoxy)phenyl]methane sulfonamide) an orally active leukotriene D 4 antagonist: Pharmacological characterization in vitro and in vivo in the guinea pig

The following communicates the pharmacology of Wy-48,252 (1,1,1-trifluoro-N-[3-(2-quinolinylmethoxy)phenyl]methanesulfonamide) a chemically novel and orally potent leukotriene (LT) D 4 receptor antagonist. In the isolated guinea-pig trachea pretreated with indomethacin (5 μM) and L-cysteine (10 mM), Wy-48,252 antagonized TD 4-induced contraction with a pK B = 7.6. Against LTC 4 on tissues pretreated with IND and glutathione (10 mM), Wy-48,252 had a pK B > 5. Wy-48,252 (10 μM) did not antagonize pilocarpine-, histamine- or PGF 2α-induced tracheal contraction. Further, in the presence of indomethacin and chlorpheniramine (1 μM), Wy-48,252 dose-dependently inhibited the antigen-induced contraction of guinea-pig trachea in a manner consistent with antagonism at the LTD 4 receptor and inhibition of LT synthesis. In the Konzett-Rossler model of i.v. LTD 4-induced bronchoconstriction in indomethacin treated guinea pigs, intragastric Wy-48,252 (2 hr) had and ID 50 of 100 μg/kg and a functionalhalf-life of 5 hr. Against i.v. antigen-induced bronchoconstriction in guinea pigs treated with indomethacin and chlorpheniramine, intragastric Wy-48,252 (2 hr) had an ID 50 of 0.6 mg/kg and a 5 hr half life. Intragastric Wy-48,252 also selectively blocked the cutaneous wheal reaction to intradermal LTD 4 but not histamine. We conclude that Wy-48,252 is distinguished from other selective LTD 4 receptor antagonists by its oral potency and should be useful in ascertaining the role of LTD 4 mediated processes in asthma, allergy and animal models.

Leukotrienes in orthodontic tooth movement

Prostaglandins (PGs) and leukotrienes (LTs) are products of arachidonic acid conversion. PGs have an established role in mediating orthodontic tooth movement. The role of LTs in modulating or mediating orthodontic tooth movement was investigated in this study. One hundred thirty-two Sprague-Dawley rats were used; the animals weighed 300 to 400 gm with equal numbers of male and female rats. They were divided into five main groups of 24 animals each and a sham group of 12 animals. An orthodontic appliance was placed and activated on all the animals except the sham group; in this group the appliances were not active. Each main group was given one of the following treatments daily: distilled water, 5% gum arabic solution, PG synthesis inhibitor indomethacin, LT synthesis inhibitor AA861, and a combination of both drugs. Each group was divided into six subgroups of four animals; the animals were killed at either 1, 3, 5, 7, 19, or 14 days, and tooth movement measured. The three sham subgroups received distilled water and were killed at 1, 7, or 10 days. The first maxillary molar (the moved tooth) and surrounding tissues were removed from all animals in the sham group and the subgroups killed at 1, 7, and 10 days in the gum arabic solution group and the LT synthesis inhibitor group. Prostaglandin E 2 (PGE 2) and leukotriene B 4 (LTB 4) were extracted, measured with radioimmunoassay (RIA), and standardized per milligram of protein in the sample. A significant inhibition of tooth movement occurred beginning on day 7 in the indomethacin, AA861, and combination groups; there was no significant difference among these groups. The leukotriene inhibitor AA861 caused a significant LTB 4 inhibition and a significant increase in PGE 2 production. These findings suggest that LTs might have a role in mediating orthodontic tooth movement.

Studies on the Mechanisms Causing Inhibition of Insulin Secretion in Rat Pancreatic Islets Exposed to Human Interleukin-1β Indicate a Perturbation in the Mitochondrial Function*

This study aimed at a more detailed characterization of the mechanisms by which interleukin 1 (IL-1) inhibits insulin secretion. For this purpose, isolated rat pancreatic islets were kept in tissue culture for 5 days in medium RPMI 1640 plus 10% calf serum. The islets were subsequently transferred to the same culture medium containing various test substances plus 1% human serum with or without 25 U/ml human recombinant IL-1 beta. After a culture period of 48 h the islet structure was examined in the electron microscope and the islet function studied in short term incubations in the absence of IL-1. Islets exposed to IL-1 showed ultrastructural signs of degeneration in 10-20% of the B cells while such changes were not found in other types of islet cells. An increased number of secondary lysosomes and occasional myelin figures were observed in the B cells exposed to IL-1. These ultrastructural alterations were, however, reversed in islets cultured in cytokine-free medium for 6 days after the IL-1 treatment. In islets cultured in the presence of 11.1 mM glucose only, or 11.1 mM glucose plus 10 mM nicotinamide, 61 mM dimethyl area, 2 micrograms/ml indomethacin, 10 microM 4-bromophenacyl bromide or 10 microM nordihydroguaiaretic acid, 10 microM phenantroline, and 0.1 or 1.0 microgram/ml cyclosporin A, IL-1 reduced the insulin release by 64-85%. Culture at 5.6 mM glucose did not modify the IL-1-induced inhibition of insulin release, whereas a significant protective effect was observed at 28 or 56 mM glucose. The DNA content in IL-1-exposed islets cultured at 11.1 mM glucose was decreased by about 20% but not in islets cultured at other glucose concentrations. The D-[5-3H]glucose utilization at 16.7 mM glucose was unaffected by IL-1, whereas the oxidation of D-[6-14C]glucose was reduced by 50%. The present results suggest that IL-1-induced inhibition of insulin secretion is related to a disturbed mitochondrial function. This effect is not counteracted by a poly(ADP-ribose) synthetase inhibitor, a hydroxyl radical scavenger, an iron chelator, a T lymphocyte-specific immunosuppressive drug, or inhibitors of phospholipase A2 or inhibitors of prostaglandin and leukotriene synthesis. Thus, IL-1-induced inhibition of insulin secretion seems not to be mediated by the same mechanisms as those causing alloxan- or streptozotocin-induced damage of B cells. Furthermore, the action of IL-1 does not appear to be mediated via arachidonic acid metabolism. Glucose affords some protection, probably by enhancing the B cell mitochondrial function.(ABSTRACT TRUNCATED AT 400 WORDS)

Activation and migration of leukocytes and vascular leakage induced by serum-opsonized zymosan particles in hamster cheek pouch

The effect of topically applied serum-opsonized zymosan (SOZ) and zymosan-activated serum (ZAS) was studied in the hamster cheek pouch. Our data suggest that early vascular leakage, evaluated by intravital fluorescence microscopy after injection of FITC-dextran, and accumulation of polymorphonuclear granulocytes (PMNGs), quantitated in fixed whole tissue specimens, induced by ZAS and SOZ were caused by complement activation, whereas the late leakages (greater than 20 min after SOZ application) was PMNG-dependent. Inhibition of prostaglandin and leukotriene synthesis by indomethacin, nordihydroguaiaretic acid (NDGA), and BW755C influence early and late vascular leakage as well as accumulation of PMNGs. These drugs also inhibited the activation of hamster PMNGs in vitro, as evaluated by chemiluminescence, in a dose-dependent manner. (-)-Terbutaline, an adrenergic beta agonist, decreased vascular leakage but had no effect on PMNG activation and accumulation.

Inhibition of leukotriene synthesis markedly accelerates healing in a rat model of inflammatory bowel disease

The role of leukotrienes in the pathogenesis of chronic colitis was investigated using a rat model. Ulceration and inflammation of the distal colon was initiated by the intracolonic administration of the hapten trinitrobenzene sulfonic acid in 50% ethanol. Leukotriene B4 synthesis increased significantly within 4 h after induction of damage, with the greatest increase observed 24–72 h after administration of the hapten. The increase in leukotriene B4 synthesis correlated well (r = 0.88) with an increase in colonic myeloperoxidase activity, a biochemical marker of neutrophil infiltration. Daily intracolonic treatment with a specific 5-lipoxygenase inhibitor, L651,392, during the first 4 days after initiation of colitis, resulted in significant reductions of colonic leukotriene B4 synthesis, colonic damage score, and colon wet weight. When examined 2 wk after initiation of colitis, the group treated with L651,392 (for the first 4 days) showed significantly less colonic damage (assessed macroscopically and histologically) and colonic inflammation (assessed histologically and by measurement of myeloperoxidase activity). The healing produced by treatment with L651,392 was comparable to that observed after treatment with 5-aminosalicylic acid in a similar manner. Although a reduction of colonic damage could be produced in this model by intracolonic pretreatment with a prostaglandin E1 analogue (rioprostil), the mechanism of action of L651,392 did not appear to be through prevention of the initial injury induced by the hapten and ethanol solution. These results demonstrate that inhibition of leukotriene synthesis results in a marked acceleration of the healing of colonic ulcers and resolution of colonic inflammation in this animal model of chronic colitis. The results are therefore consistent with the hypothesis that leukotrienes play an important role in the pathogenesis of intestinal inflammation.

Angiotensin increases arachidonate metabolism in cultured anterior pituitary cells

Angiotensin peptides produced a rapid (within 30 seconds) and concentration-dependent increase in the levels of the unesterified fatty acid and stimulated prolactin release. [3H]arachidonate appeared to be cleaved primarily from the lipid of the phosphoinositide cycle. In fact, angiotensin II produced a significant reduction of [3H]phosphatidylinositol. Conversely, [3H]diacylglycerol rapidly increased after the addition of angiotensin II to the incubation medium, returned to basal values within 2 minutes and then decreased below the control levels by 3-4 minutes, possibly due to the release of [3H]arachidonate. Finally, RHC 80267, a rather selective inhibitor of diglyceride lipase, antagonized angiotensin II-induced [3H]arachidonic acid and prolactin release. The effect of arachidonate on prolactin release has to be ascribed to further metabolism of the fatty acid to products other than prostaglandins and thromboxanes. The inhibitor of the cyclo-oxygenase pathway, indomethacin, did not significantly modify angiotensin II-induced prolactin release, whereas BW 755c and ETYA (inhibitors of cyclo-, lipo- and epoxygenase pathways) and NDGA (an inhibitor of leukotriene and epoxyeicosanoid synthesis) completely counteracted the effect of the octapeptide on hormone release.

The Contrasting Influence of Two Lipoxygenase Inhibitors on Hypoxic Pulmonary Vasoconstriction in Anesthetized Pigs

Because leukotrienes may mediate hypoxic pulmonary vasoconstriction (HPV), we examined the influence of two lipoxygenase inhibitors on HPV in anesthetized pigs. HPV was induced by ventilation with a hypoxic gas mixture (FIO2 at 0.095), resulting in a fall in PaO2 to 23 +/- 2 mm Hg and a rise in pulmonary vascular resistance from 285 +/- 15 to 595 +/- 30 dyne/s/cm-5. After infusion of either U-60,257B (50 mg/kg, n = 13) or BW 755c (20 mg/kg, n = 8), the responses to repeated hypoxic challenges were recorded. After U-60,257B infusion the hypoxic pressor response was eliminated at 10 and 30 min and remained significantly (p less than 0.01) attenuated at 50 min. The pulmonary pressor response to angiotensin II infusion (0.2 micrograms/kg/min) was also ablated, whereas the systemic response was unchanged. In contrast, after BW 755c infusion there was a modest but sustained augmentation of HPV, maximum at 30 min (pulmonary vascular resistance, 158 +/- 23% control, p less than 0.01), and no alteration of the responses to angiotensin II. BW 755c inhibited the A23187-induced release of leukotrienes, but not histamine, from isolated porcine lung cells (IC50, 6.3 x 10(-5) M), whereas U-60,257B inhibited the release of both leukotriene (IC50, 1.1 x 10(-4) M) and histamine. These findings indicate that reduction of HPV by lipoxygenase inhibitors is not necessarily a consequence of inhibition of leukotriene synthesis.

Tolfenamic acid inhibits leukotriene synthesis in human granulocytes

Higher concentrations of TA were needed to inhibit chemiluminescence than LTB4 formation (whereas indomethacin and timegadine reduced these two phenomena with similar potencies) suggesting a specific blockade of 5-lipoxygenase by TA. Oral TA decreased leukotriene production in PMNsex vivo. This may be related to the good anti-inflammatory and anti-migraine actions of TA, and partly explain its rare gastric and bronchoconstrictive adverse effects. However, the recommended single oral dose (100–400 mg) seems to be too low to decrease systemic LT-synthesis in all subjects.

Hydrogen peroxide inhibits alveolar macrophage 5-lipoxygenase metabolism in association with depletion of ATP.

We have previously shown that the biologically important reactive oxygen metabolite hydrogen peroxide (H2O2) stimulates arachidonic acid (AA) release and thromboxane A2 synthesis in the rat alveolar macrophage. We have now investigated the effects of H2O2 on alveolar macrophage 5-lipoxygenase metabolism. H2O2 failed to stimulate detectable synthesis of leukotriene B4, leukotriene C4, or 5-hydroxyeicosatetraenoic acid (5-HETE) as determined by reverse-phase high performance liquid chromatography (RP-HPLC) and sensitive radioimmunoassays (RIAs). This was not explained by oxidative degradation of leukotrienes by H2O2 at the concentrations used. Moreover, RIA and RP-HPLC analyses demonstrated that H2O2 dose-dependently inhibited synthesis of leukotriene B4, leukotriene C4, and 5-HETE induced by the agonists A23187 (10 microM) and zymosan (100 micrograms/ml), over the same concentration range at which it augmented synthesis of the cyclooxygenase products thromboxane A2 and 12-hydroxy-5,8,10-heptadecatrienoic acid. Four lines of evidence suggested that H2O2 inhibited alveolar macrophage leukotriene and 5-HETE synthesis by depleting cellular ATP, a cofactor for 5-lipoxygenase. 1) H2O2 depleted ATP in A23187- and zymosan-stimulated alveolar macrophages with a dose dependence very similar to that for inhibition of agonist-induced leukotriene synthesis. 2) The time courses of ATP depletion and inhibition of leukotriene B4 synthesis by H2O2 were compatible with a rate-limiting effect of ATP on leukotriene synthesis in H2O2-exposed cultures. 3) Treatment of alveolar macrophages with the electron transport inhibitor antimycin A prior to A23187 stimulation depleted ATP and inhibited leukotriene B4 and C4 synthesis to equivalent degrees, while thromboxane A2 production was spared. 4) Incubation with the ATP precursors inosine plus phosphate attenuated both ATP depletion and inhibition of leukotriene B4 and C4 synthesis in alveolar macrophages stimulated with A23187 in the presence of H2O2. Our results show that H2O2 has the capacity to act both as an agonist for macrophage AA metabolism, and as a selective inhibitor of the 5-lipoxygenase pathway, probably as a result of its ability to deplete ATP. Depletion of cellular energy stores by oxidants generated during inflammation in vivo may be a means by which the inflammatory response is self-limited.

Preferential inhibition of 5-lipoxygenase activity by manoalide

Treatment of human polymorphonuclear leukocytes (PMNLs) with mumolar concentrations of the anti-inflammatory drug manoalide inhibited production of leukotriene B 4 (LTB 4) and LTC 4/LTD 4 in response to the calcium ionophore A23187. In an attempt to further define the mechanism(s) of action of this agent, we have examined its interaction with several lipoxygenase enzymes. In RBL-1 cells, manoalide inhibited 5-lipoxygenase (5-LO) activity with an approximate IC 50 of 0.3 μM. This was equipotent in our system with the known lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA). Manoalide was virtually inactive, however, against 12-lipoxygenase activity in both human platelets and mouse epidermis, with little inhibition seen at concentrations up to 100 μM. Manoalide showed some activity against soybean lipoxygenase, although it was 30- to 50-fold less potent than as an inhibitor of the 5-lipoxygenase enzyme. These data indicate that manoalide is a selective 5-LO inhibitor and suggest the possibility that its anti-inflammatory actions may be due, at least in part, to inhibition of leukotriene synthesis.