calcimycin and nimesulide

1. Cyclo-oxygenase (COX) and lipoxygenase (LO) share a common substrate, arachidonic acid. Aspirin and related drugs inhibit COX activity. In a subset of patients with asthma aspirin induces clinical symptoms associated with increased levels of certain LO products, a phenomenon known as aspirin-sensitive asthma. The pharmacological pathways regulating such responses are not known. 2. Here COX-1 and LO activity were measured respectively by the formation of thromboxane B(2) (TXB(2)) or leukotrienes (LT) C(4), D(4) and E(4) in whole blood stimulated with A23187. COX-2 activity was measured by the formation of prostaglandin E(2) (PGE(2)) in blood stimulated with lipopolysaccharide (LPS) for 18 h. 3. No differences in the levels of COX-1, COX-2 or LO products or the potency of drugs were found in blood from aspirin sensitive vs aspirin tolerant patients. Aspirin, indomethacin and nimesulide inhibited COX-1 activity, without altering LO activity. Indomethacin, nimesulide and the COX-2 selective inhibitor DFP [5,5-dimethyl-3-(2-isopropoxy)-4-(4-methanesulfonylphenyl)-2(5H)-furanone] inhibited COX-2 activity. NO-aspirin, like aspirin inhibited COX-1 activity in blood from both groups. However, NO-aspirin also reduced LO activity in the blood from both patient groups. Sodium salicylate was an ineffective inhibitor of COX-1, COX-2 or LO activity in blood from both aspirin-sensitive and tolerant patients. 4. Thus, when COX activity in the blood of aspirin-sensitive asthmatics is blocked there is no associated increase in LO products. Moreover, NO-aspirin, unlike other NSAIDs tested, inhibited LO activity in the blood from both aspirin sensitive and aspirin tolerant individuals. This suggests that NO-aspirin may be better tolerated than aspirin by aspirin-sensitive asthmatics.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Aspirin; Asthma; Benzene Derivatives; Calcimycin; Cyclooxygenase 1; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Cyclooxygenase Inhibitors; Dinoprostone; Dose-Response Relationship, Drug; Furans; Humans; Indomethacin; Ionophores; Isoenzymes; Leukotrienes; Lipopolysaccharides; Lipoxygenase; Membrane Proteins; Prostaglandin-Endoperoxide Synthases; Sulfonamides; Thromboxane B2

Interleukin (IL)-1beta impairs human airway smooth muscle (ASM) cell cAMP responses to isoproterenol (Iso). We investigated if bradykinin (BK) could cause a similar effect and the role of cyclooxygenase (COX) products in this event, since we have recently reported that BK, like IL-1beta, also causes COX-2 induction and prostanoid release in human ASM cells. BK pretreatment significantly attenuated Iso-induced cAMP generation in a time- and concentration-dependent manner. cAMP generation by prostaglandin (PG) E2 but not by forskolin was also impaired. The COX inhibitor indomethacin completely prevented the impairment, whereas the selective COX-2 inhibitors NS-398 and nimesulide, protein synthesis inhibitors cycloheximide and actinomycin D, and steroid dexamethasone were all partially effective. The impairment was mimicked by the B2 agonist [Tyr(Me)8]BK, the Ca2+ ionophore A-23187, and PGE2 and prevented by the B2 antagonist HOE-140, but anti-IL-1beta serum was ineffective. The results indicate that BK impairs human ASM cell responses to Iso, and the effect is largely mediated by B2 receptor-related COX product release via both COX isoforms and is independent of IL-1beta.

Topics: Adult; Bradykinin; Calcimycin; Cells, Cultured; Cyclic AMP; Cycloheximide; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Cyclooxygenase Inhibitors; Dactinomycin; Dexamethasone; Dinoprostone; Enzyme Induction; Female; Humans; Indomethacin; Isoenzymes; Isoproterenol; Male; Membrane Proteins; Middle Aged; Muscle, Smooth; Nitrobenzenes; Prostaglandin-Endoperoxide Synthases; Receptor, Bradykinin B2; Receptors, Bradykinin; Sulfonamides; Trachea

Nimesulide (NIM) is a sulfonanilide nonsteroidal anti-inflammatory drug (NSAID) used in the treatment of various inflammatory diseases and chemically unrelated to other acidic NSAIDs, such as acetylsalicylic acid (ASA) and indomethacin (INDO). We investigated the effects of NIM and of its in vivo metabolite, 4-hydroxy-NIM (OH-NIM), on the release of performed (histamine) and de novo synthesized mediators (sulfidopeptide leukotriene C4 [LTC4] and prostaglandin D2 [PGD2]) from human basophils and mast cells isolated from lung parenchyma (HLMC) and skin (HSMC). Histamine release from basophils challenged with rabbit anti-human IgE antibody (anti-IgE) was enhanced by preincubation with ASA or INDO (92.2 +/- 7.1% at 10(-3) M and 61.1 +/- 6.7% at 3 x 10(-6) M, respectively; P < .001). In contrast, NIM and its metabolite, OH-NIM (10(-6) to 10(-3) M), caused concentration-dependent inhibition (2.9 to approximately 60% and 3.7 to approximately 90%, respectively) of IgE-mediated histamine release from basophils. NIM and OH-NIM also inhibited histamine release from basophils induced by the Ca++ ionophore A23187 and different protein kinase C activators, such as 12-O-tetradecanoyl-phorbol-13-acetate, bryostatin 1 and bryostatin 5. NIM and OH-NIM also inhibited the IgE-mediated histamine release from HLMC (52.3 +/- 9.6% and 66.1 +/- 12.1% at 10(-3) M, respectively; P < .0001) and HSMC (67.3 +/- 3.7% and 77.7 +/- 12.0% at 10(-3) M, respectively; P < .0001) but had little or no effect on HLMC and HSMC activated by A23187. NIM (10(-6) to 10(-3) M) markedly inhibited the de novo synthesis of LTC4 from basophils, LTC4 and PGD2 from HLMC and PGD2 from HSMC. NIM and OH-NIM potentiated, whereas ASA and INDO reversed, the inhibitory effect of adenylate cyclase agonists, such as prostaglandin E1 and forskolin. In addition, NIM and OH-NIM reversed the enhancing effects of ASA and INDO on IgE-mediated histamine release from basophils.

Topics: Adolescent; Adult; Alprostadil; Amino Acid Sequence; Anti-Inflammatory Agents, Non-Steroidal; Aspirin; Basophils; Bryostatins; Calcimycin; Colforsin; Complement C5a; Histamine Release; Humans; Immunoglobulin E; Indomethacin; Lactones; Leukotriene C4; Lung; Macrolides; Mast Cells; Middle Aged; Molecular Sequence Data; N-Formylmethionine Leucyl-Phenylalanine; Prostaglandin D2; Skin; Sulfonamides; Tetradecanoylphorbol Acetate

Nimesulide (CAS 51803-78-2) is a methane sulphoanilide derivative provided with specific anti-inflammatory activity. In human polymorphonuclear leucocytes (PMNs), the activity of nimesulide has been suggested to be based on the inhibition of the oxidative burst. However, the effect of the compound on PMNs function seems to be very complex. In order to give a major insight into the mechanism of action of nimesulide, the effect of the drug was studied in vitro on human PMNs functions, such as free radical generation and enzyme release, and on cytosolic free calcium levels, following the activation with specific stimuli. Moreover, the hypothesis that nimesulide could act by interfering with the adenosine cell receptor system was also evaluated. Nimesulide (1-50 mumol/l showed a dose-dependent inhibitory activity on superoxide anion and chemiluminescence production from PMNs stimulated with the oligopeptide fMLP, the ionophore A23187, and the phorbol ester PMA. Enzyme release was significantly reduced, when fMLP and A23187 represented the stimulating agents, while no effect at all was observed with PMA. Studies with the fluorescent calcium chelating dye FURA 2/AM showed that nimesulide was able to reduce free cytosolic calcium increase produced by fMLP and the ionophore ionomycin. The preincubation of cells with the specific adenosine receptor antagonist theophylline was able to significantly reverse the inhibitory activity of nimesulide, either on free radical production and enzyme release, and on free cytosolic calcium increase sustained by fMLP and the ionophores.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenosine; Anti-Inflammatory Agents, Non-Steroidal; Calcimycin; Calcium; Free Radicals; Glucuronidase; Humans; In Vitro Techniques; Luminescent Measurements; Muramidase; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Receptors, Purinergic P1; Sulfonamides; Superoxides; Theophylline