Effects of some non-steroidal anti-inflammatory drug copper complexes on polymorphonuclear leukocyte oxidative metabolism

Interaction between anti-inflammatory drugs and reactive oxygen metabolites must be considered in the course of pharmacological studies intended to develop new compounds. Effects of indomethacin, aspirin, and 3,5-diisopropylsalicylic acid (3,5-DIPS) and their copper complexes on PMNL oxidative metabolism and the evolution of an acute inflammatory reaction were studied in the rat. Experiments were performed in vitro by assessment of superoxide generation and reduction of chemiluminescence by PMNLs incubated or not (control) in medium containing various concentrations of these compounds. A dose-related decrease of these parameters was observed, however, copper complexes were found to be more effective than their parent drugs or Cu gluconate. Copper complexes were also more effective anti-inflammatory agents than their parent ligands or Cu gluconate when the volume of exudate and number of exudate PMNLs were assessed after induction of pleurisy in rats by injection of isologous serum. It is concluded that modulation of the PMNL oxidative burst by copper complexes offers an accounting for the anti-inflammatory activity of these compounds.     

Effect of antituberculous drugs on human polymorphonuclear leukocyte functions in vitro

The aim of the study was to investigate antituberculous drugs effects on polymorphonuclear leukocyte (PMN) functions (phagocytic activity and intracellular killing activity) in vitro. PMNs obtained from healthy volunteers were incubated with antituberculous drugs (isoniazid [INH], rifampin [RIF], pyrazinamide [PZA], ethambutol [EMB], streptomycin [S], amikacin [A], ofloxacin [OFLX], prothionamide [PTH] and cycloserine [CyC]) and different combinations at therapeutic serum concentrations. Phagocytic activity of PMNs was significantly increased when compared with controls by PTH (p<0.001), A (p<0.001), OFLX (p<0.001), INH+RIF+S combination (p<0.01), A+OFLX combination (p<0.05), A+OFLX+CyC combination (p<0.01) and A+OFLX+CyC+PTH+EMB combination (p<0.01). Intracellular killing activity of PMNs was significantly increased by OFLX when compared with the control (p<0.05). No significant difference was observed in functions of PMN for other drugs when compared with control (p>0.05). Functions of PMN were significantly increased by OFLX when compared with A+OFLX combination (p<0.05). Phagocytic activity of PMNs was significantly increased by A+OFLX+CyC combination and A+OFLX+CyC+PTH+EMB combination when compared with A+OFLX+CyC+PTH combination and A+OFLX+CyC+PTH+PZA combination (p<0.05). No significant difference was found in functions of PMN between the other groups (p>0.05). In conclusion, some antituberculous drugs alone or in combination enhanced PMN functions, although in combination no additive or synergistic effects were detected. Moreover, none of the antituberculous drugs alone or in combination significantly decreased PMN functions. The drugs having adverse effects on immune functions would better be replaced with equally effective drugs or drug combinations having positive effects on PMN functions.     

Effects of non-steroidal anti-inflammatory agents on human neutrophil functions in vitro and in vivo

Human blood neutrophils exposed to appropriate stimuli aggregate, degranulate and generate superoxide anion (O2-). These responses are anteceded by mobilization of membrane-associated calcium, monitored as a decrease in fluorescence of cells preloaded with chlortetracycline (CTC). We studied the effects, both in vitro and in vivo, of non-steroidal anti-inflammatory agents (aspirin, indomethacin, ibuprofen and piroxicam) on these neutrophil responses to three stimuli: a chemoattractant, N-formyl-methionyl-leucyl-phenylalanine (FMLP); a tumor promotor, phorbol myristate acetate (PMA); and a lectin, concanavalin A (Con A). The effects of these drugs were compared with those of two polyenoic inhibitors of arachidonate metabolism: eicosatrienoic acid (ETI) and eicosatetraynoic acid (ETYA). The pattern of inhibition of neutrophil functions varied both with inhibitor and the nature of the stimulus. Thus, aspirin, piroxicam, ETYA and ETI inhibited neutrophil aggregation, degranulation, and O2- generation in response to FMLP, whereas ibuprofen inhibited only aggregation and degranulation and indomethacin only inhibited aggregation. None of the agents inhibited aggregation or degranulation induced by PMA or Con A: only piroxicam inhibited O2- generation in response to PMA or Con A. ETI and ibuprofen inhibited decrements of CTC fluorescence induced by FMLP, but whereas ETI inhibited the CTC response to PMA or Con A, ibuprofen was without effect. The agents had varying effects on binding of the stimulus [( 3H]FMLP, [3H]Con A), but these did not correlate with neutrophil responses to the ligands. Neutrophils from subjects taking therapeutic doses of ibuprofen, indomethacin, or piroxicam showed profiles of inhibited responses to FMLP similar to those observed with these agents in vitro. These data suggest that, although non-steroidal anti-inflammatory agents may inhibit discrete neutrophil functions both in vitro and in vivo, their effects do not duplicate those of polyenoic inhibitors of arachidonate metabolism. Moreover, since the susceptibility of neutrophils differed not only with respect to each inhibitor, but also to the stimulus, it is unlikely that all neutrophil responses are necessarily linked by a common pathway that is blocked by inhibitors of arachidonic acid metabolism.     

In vitro effects of non-steroidal anti-inflammatory drugs on human polymorphonuclear cells and lymphocyte migration.

1. The action of the non-steroidal anti-inflammatory drugs (NSAID), sodium salicylate, aspirin, phenylbutazone and indomethacin was investigated on the migration of human polymorphonuclear cells (PMNs) and lymphocytes, using the system of migration of leucocytes from glass capillary tubes. 2. All NSAID produced a dose-dependent inhibition of cell migration, and were more effective on the migration of the PMN than on lymphocytes. 3. Drugs optimally suppressed PMN migration after 20 to 24 h incubation, and lymphocytes after 3 to 6 h. 4. Prolonged incubation of cells with several concentrations of NSAID demonstrated an 'escape' from inhibition in PMNs prepared from one subject.     

我院门诊非甾体抗炎药的合理使用情况调查

目的：通过调查我院门诊处方中非甾体抗炎药的使用情况,并对调查结果进行分析和讨论,促进我院非甾体抗炎药使用的合理化和规范化。方法：随机抽取2008年01月至2008年12月的21223张门诊处方中非甾体抗炎药的使用情况进行统计,按使用频率进行排序,取前5位药物,分析其在年龄、使用剂量的合理性和该药物所用的症状-9药物说明书的适应证的相符性方面的临床分布特点。结果：服用非甾体抗炎药患者1564例,占门诊处方总数21223例的7．37％。使用频率前5位的非甾体抗炎药分别为阿司匹林肠溶片、二氟尼柳片、美洛昔康片、复方氯唑沙宗片、双氯芬酸缓释片,统计药物的DUI值大多符合规定,其中使用频率前5位的药物为100％合理使用（DuI值等于或小于1）。使用频率前5位的非甾体抗炎药总相符率为75．7％。病种以损伤史最多,其次是关节炎、腰背痛,分别为：167例（10．78％）、160例（10．23％）、154例（9．85％）。结论：我院非甾体抗炎药的使用与说明书适应证的相符率较高,该类药物的使用基本合理。     

Effects of acetaminophen and nonsteroidal anti-inflammatory drugs on progesterone production by porcine granulosa cells in vitro

We investigated the effects of a group of pharmaceutical agents commonly ingested by reproductive-aged women, acetaminophen and the nonsteroidal anti-inflammatory drugs (NSAID), on progesterone (P) production by cultures of highly differentiated porcine granulosa cells. These compounds were added to cultures over a dose range of 10⁻⁸ to 10⁻⁵ M and P, and cell protein was measured after 24 hours. P production was suppressed by acetaminophen, fenoprofen, and sulindac to a maximum of 81%, 76%, and 71% of control, respectively. P production was enhanced by butazolidin at all doses tested to a maximum of 140% of control. Granulosa cell protein was suppressed by butazolidin and salicylic acid to a maximum of 81% of controls. These data imply that acetaminophen and several NSAID have the potential for clinical reproductive toxicity with differing individual effects on reproductive tract tissues, suggesting further selective testing in vivo.     

Effects of prior use of non-steroidal anti-inflammatory drugs on renal function and transfusion requirements after upper gastrointestinal haemorrhage

Summary The possibility has been investigated that, after admission to hospital with acute upper gastrointestinal bleeding, patients who have been users of aspirin and non-aspirin non-steroidal anti-inflammatory drugs have poorer baseline renal. function, a greater improvement in renal function during their hospital stay, and a larger transfusion requirement than non-users. Patients over 50 years of age admitted to public hospitals with acute upper gastrointestinal bleeding were studied. Creatinine clearance was estimated from serum creatinine and the transfusion requirement was recorded as the number of units of blood transfused on Day 1 and throughout the entire hospital stay. Data were obtained prospectively from case notes and by structured interview.Users of non-steroidal anti-inflammatory drugs were significantly older than non-users. The estimated creatinine clearance on admission to hospital declined with age. Creatinine clearance was 13.2 (95 % CI 6.0 to 20.4) ml · min' lower in users than non-users of non-aspirin non-steroidal anti-inflammatory drugs. However, the difference was attributable to the older age of the drug users rather than to the drugs themselves.On average, the increase in creatinine clearance during hospital stay was the same in users and non-users of non-aspirin non-steroidal anti-inflammatory drugs. Prior use of aspirin had no effect on any measure of renal function. The incidence of blood transfusion was higher in older than in younger patients but neither the incidence of transfusion, nor the transfusion requirement, was different between users and non-users of non-aspirin non-steroidal anti-inflammatory drugs and aspirin.Although the study has not excluded certain adverse effects of prior use of these drugs on renal and haemostatic function after an episode of upper gastrointestinal bleeding, it does indicate that such change are unlikely to be of major clinical significance.     

Influence of prolonged exposure of a short half life non-steroidal anti-inflammatory drugs on gastrointestinal safety

Aims  To test the influence of frequent concentration peaking, as occurs in multiple-dosing of non-steroidal anti-inflammatory drugs (NSAIDs) with short t _1/2, and duration of therapy of NSAIDs on gastrointestinal permeability. Methodology  2.5 mg/(kg 12 h) flurbiprofen was administered as repeated oral and interperitoneal (i.p) doses or as i.p. osmotic pump (once implanted to mimic long t _1/2) for 7 days to healthy rats. Urinary excretion of ⁵¹Cr-EDTA (days 0, 1, 4 and 7 during all regimens) and sucrose (days 0, 1 and 7 for i.p. doses) were measured as markers of gastroduodenal and intestinal permeability, respectively. Results  Both i.p. regimens elevated ⁵¹Cr-EDTA permeability suggestive of a systemic effect. There was no significant difference between the i.p regimens in ⁵¹Cr-EDTA permeability. The first day ⁵¹Cr-EDTA permeability was significantly higher for the oral than for the i.p. doses suggestive of a topcal effect. The effect became less potent with time despite continuous dosing indicating adaptation for both topical and systemic effects. None of the i.p. regimen altered sucrose permeability. Conclusion  NSAID’s potency to increase permeability reduces with time despite continuous dosing. Topical effect following oral dosing, and not the frequent peaking differentiates regimens from each other in elevating ⁵¹Cr-EDTA permeability. The repeated dosing rather than the magnitude of t _1/2 may influence the gut safety profile of NSAIDs.     

Inhibitory effects of non-steroidal anti-inflammatory drugs on human liver microsomal morphine glucuronidation: Implications for drug-drug interaction liability

The inhibitory effects of fifteen NSAIDs from six structurally distinct classes on human liver microsomal morphine glucuronidation were investigated. K_i values of selected NSAIDs were generated and employed to assess DDI liability in vivo. Potent inhibition was observed for mefenamic acid and tolfenamic acid; respective IC₅₀ values for morphine 3- and 6-glucuronidation were 9.2 and 13.5 μM, and 5.3 and 8.3 μM. Diclofenac and celecoxib showed moderate inhibition with IC₅₀ values of 78 and 52 μM, and 83 and 214 μM, respectively. Estimated IC₅₀ values for the other NSAIDs screened were >100 μM. Mefenamic acid, diclofenac, and S-naproxen competitively inhibited morphine 3- and 6-glucuronidation, with the K_i values of 11 and 12 μM, 110 and 76 μM, and 319 and 650 μM, respectively. Using the static mechanistic IVIVE approach, an approximate 40% increase in the AUC of morphine was predicted when co-administered with mefenamic acid, whereas the increase was <10% with diclofenac and S-naproxen. PBPK modeling predicted <15% increases in the morphine AUC from diclofenac and S-naproxen inhibition in virtual healthy and cirrhotic subjects. The data suggest that potential clinically significant DDIs arising from NSAID inhibition of morphine glucuronidation is unlikely, with the possible exception of some fenamates.     

山茶油对非甾体抗炎药经皮渗透的促进作用

目的考察山茶油对5种非甾体抗炎药体外经皮渗透的促进作用及其与药物油水分配系数的关系。方法采用两室扩散池装置,以离体大鼠腹部皮肤为渗透屏障,用预处理皮肤法考察山茶油的促渗透作用;用HPLC法测定非甾体抗炎药不同时间在接受池药物浓度,计算累积渗透量及其他渗透动力学参数;用摇瓶法测定药物正辛醇介质分配系数以拟合其与山茶油促渗活性关系。结果山茶油可有效促进非甾体抗炎药的经皮渗透率(除双氯芬酸钠外),尤其对氟比洛芬,促渗效果最明显(P<0.01),其增渗倍数达到3.51;山茶油的促渗活性与非甾体抗炎药的正辛醇介质分配系数呈类似抛物线关系。结论山茶油作为一种低毒、无刺激性的渗透促进剂,能够有效提高药物的经皮渗透效果,具有良好的开发应用前景。     

Role of carbonyl reducing enzymes in the phase I biotransformation of the non-steroidal anti-inflammatory drug nabumetone in vitro

1.?Nabumetone is a clinically used non-steroidal anti-inflammatory drug, its biotransformation includes major active metabolite 6-methoxy-2-naphtylacetic acid and another three phase I as well as corresponding phase II metabolites which are regarded as inactive. One important biotransformation pathway is carbonyl reduction, which leads to the phase I metabolite, reduced nabumetone.

2.?The aim of this study is the determination of the role of a particular human liver subcellular fraction in the nabumetone reduction and the identification of participating carbonyl reducing enzymes along with their stereospecificities.

3.?Both subcellular fractions take part in the carbonyl reduction of nabumetone and the reduction is at least in vitro the main biotransformation pathway. The activities of eight cytosolic carbonyl reducing enzymes – CBR1, CBR3, AKR1B1, AKR1B10, AKR1C1-4 – toward nabumetone were tested. Except for CBR3, all tested reductases transform nabumetone to its reduced metabolite. AKR1C4 and AKR1C3 have the highest intrinsic clearances.

4.?The stereospecificity of the majority of the tested enzymes is shifted to the production of an (+)-enantiomer of reduced nabumetone; only AKR1C1 and AKR1C4 produce predominantly an (?)-enantiomer. This project provides for the first time evidence that seven specific carbonyl reducing enzymes participate in nabumetone metabolism.     

Inhibiting effect of non-steroidal anti-inflammatory drugs on the chemotaxis of human leukocytes in vitro.

Chemotaxis of human leukocytes was inhibited in vitro by four non-steroidal anti-inflammatory drugs, namely, lysine acetylsalicylate, phenylbutazone, indomethacin, and indoprofen. Dose-dependent effects were always found and significant regression was proved for all drugs except phenylbutazone.     

Effects of non-steroidal antiinflammatory drugs on D-serine-induced oxidative stress in vitro

Inflammation is deleterious for organs with reduced capacity of regeneration, such as the brain. Recently, studies have focused on investigating the therapeutic effects of nonsteroidal anti-inflammatory drugs (NSAIDs) in Alzheimer's disease, Parkinson's disease, Huntington's disease, and multiple sclerosis. Excitotoxicity is the pathological process when receptors for the excitatory neurotransmitter glutamate, such as the N-methyl-D-aspartate (NMDA), receptors are overactivated. This process may be involved in neurodegenerative diseases. D-serine is one of the coagonist of NMDA receptors, and increased levels of D-serine are associated with excitotoxicity. In our study, the potential neuroprotective effects of mefenamic acid, acetaminophen, and naproxen sodium were investigated against D-serine-induced oxidative stress in the rat brain in vitro. To show their potential neuroprotective properties, NSAIDs were incubated with D-serine and reactive oxygen species (ROS), malondialdehyde, and protein carbonyl content of the brain after different treatments were measured. Our results demostrate that NSAIDs used in the present study significantly reduced ROS production, lipid peroxidation, and protein oxidation against D-serine treatment.     

Effects of non-steroidal anti-inflammatory drugs on the pharmacokinetics and elimination of aciclovir in rats

This study aims to investigate the effect of commonly used non-steroidal anti-inflammatory drugs (NSAIDs) on the pharmacokinetics and the renal elimination of aciclovir in rats. Pharmacokinetic parameters were determined following an intravenous administration of aciclovir (5 mg kg(-1)) to rats in the presence and absence of ketoprofen or naproxen (25 mg kg(-1)). Compared with the control (given aciclovir alone), pre-treatment with ketoprofen or naproxen 30 min before aciclovir administration significantly altered the pharmacokinetics of aciclovir. Renal clearance of aciclovir was reduced by approximately two fold in the presence of ketoprofen or naproxen. Consequently, the systemic exposure (AUC) to aciclovir in the rats pre-treated with ketoprofen or naproxen was significantly (P < 0.05) higher than that from the control group given aciclovir alone. Furthermore, the mean terminal plasma half-life of aciclovir was enhanced by 4-5 fold by pre-treatment with ketoprofen or naproxen. These results suggest that NSAIDs, such as ketoprofen and naproxen, are effective in altering the pharmacokinetics of aciclovir by inhibiting the organic anion transporter-mediated tubular secretion of aciclovir. Therefore, concomitant use of ketoprofen or naproxen with aciclovir should require close monitoring for clinical consequence of potential drug interaction.     

Effects of lead on polymorphonuclear leukocyte (PMN) functions in occupationally exposed workers

Previous in vitro experiments have shown that lead can inhibit PMN chemotaxis, phagocytosis and super-oxide formation. Moreover, we have observed an inhibition of PMN chemotaxis in workers occupationally exposed to lead with a mean blood lead concentration of 3.06 (mol/l. The present study was carried out to evaluate locomotion and luminol assisted chemiluminescence (CL) of polymorphonuclear leukocytes (PMN) harvested from ten lead occupationally exposed workers with blood lead concentrations of 1.59 mol/l (SD 0.27 mol/l). Since lipids affect PMN activity and lipid composition is modified in erythrocytes of lead workers, PMN lipids were also studied. Ten healthy male subjects of the same age were taken as controls. Chemotaxis, i.e. locomotion stimulated through a specific membrane receptor, was impaired in the PMN of lead workers, but random migration, i. e. unstimulated cell locomotion, and respiratory burst were both unmodified. Cholesterol and phospholipids were not changed, but the percentage of arachidonic acid was significantly increased. The release of LTB4, generated by the oxidative metabolism of arachidonic acid, was increased. CL, which detects reactive oxygen species (ROS), was unmodified, but this lack of change could be the result of an increase in ROS, due to the augmentated percentage of arachidonic acid, and of a decrease in ROS, due to a direct inhibitory effect of lead on ROS generation. On the basis of the results from these ex vivo experiments, the conclusion that chemotaxis is the PMN function primarily affected by lead was confirmed. PMN are considered to be one of the first cellular targets for the action of lead; low exposure to lead modifies their activity and mainly modifies chemotaxis and LTB4 production.     

In vitro inhibitory effects of non-steroidal antiinflammatory drugs on UDP-glucuronosyltransferase 1A1-catalysed estradiol 3beta-glucuronidation in human liver microsomes

The inhibitory potencies of non-steroidal antiinflammatory drugs (NSAID) on UDP-glucuronosyltransferase (UGT) 1A1-catalysed estradiol 3beta-glucuronidation (E3G) were investigated in human liver microsomes (HLM). Inhibitory effects of the following seven NSAID were investigated: acetaminophen, diclofenac, diflunisal, indomethacin, ketoprofen, naproxen and niflumic acid. Niflumic acid had the most potent inhibitory effect on E3G with an IC50 value of 22.2 microM in HLM. The IC50 values of diclofenac, diflunisal, indomethacin for E3G were 60.9, 37.8 and 51.5 microM, respectively, while acetaminophen, ketoprofen and naproxen showed less potent inhibition. Diclofenac inhibited E3G non-competitively with a Ki value of 112 microM in HLM. The IC50 value of diclofenac for 4-methylumbelliferone glucuronidation in recombinant human UGT1A1 was 57.5 microM, similar to that obtained for E3G using HLM.In conclusion, niflumic acid had the most potent inhibitory effects on UGT1A1-catalysed E3G in HLM among seven NSAID investigated.     

Human whole blood assay for rapid and routine testing of non-steroidal anti-inflammatory drugs (NSAIDs) on cyclo-oxygenase-2 activity

Aim of this study was to present a simple, fast and reliable method to examine the capacity of NSAIDs to inhibiting COX-2 activity that uses rapid (stimulation takes only 5 h compared to other existing protocols) and routine testing. The assay includes elimination of COX-1-activity using ASS (a selective COX-1 inhibitor) and the thromboxane synthetase inhibitor (TXBSI), COX-2 induction via LPS and measurement of PGE(2). Using TXBSI reduces the amount of LPS and results in higher prostaglandin production.Cremophor EL((R))-EtOH was used as vehicle instead of DMSO because within a defined concentration range, Cremophor EL((R))-EtOH allows even very hydrophobic drugs to be solubilized and applied in vitro without cell damage. Cremophor EL((R))-EtOH at 0.2% was optimal as at this relatively low concentration excellent drug dissolution was obtained whereas many hydrophobic substances precipitate in 0.2% DMSO.Our results demonstrate that the IC(50) values for the tested NSAIDs are in the range of published data.