Inhibition of human phenol and estrogen sulfotransferase by certain non-steroidal anti-inflammatory agents

We hypothesized that aryl acetate- and aryl carboxylate-containing drugs would inhibit human phenol sulfotransferase (SULT1A1), and that selectivity would depend upon the interaction of the aryl portion of the molecule with the sulfotransferase acceptor binding site. This hypothesis was based on results with the rat orthologue showing that oxidation of phenolic substrates to carboxylate derivatives resulted in competitive inhibition of rat phenol sulfotransferase. We chose nine structurally representative non-steroidal anti-inflammatory agents and determined their inhibitory potency and selectivity toward SULT1A1 and expressed human estrogen sulfotransferase (SULT1E1). The results show that the tested agents reversibly inhibit SULT1A1 activity with IC(50) ranging from 0.1 microM to 3800 microM. These agents also inhibited SULT1E1 (IC(50) = 6 microM to 9000 microM). The agents were clearly isoform selective, with IC(50) ratios (1E1/1A1) ranging from 0.01 to 200. Nimesulide, meclofenamate, and piroxicam were more selective towards SULT1A1 inhibition, while sulindac and ibuprofen were more selective towards SULT1E1 inhibition. Sulfotransferase inhibition was maintained after substituting the carboxylate with enolate (nimesulide) or methylsulfonamide (piroxicam). Kinetic studies determined the type of inhibition of SULT1A1 for three agents (meclofenamate, nimesulide, aspirin) to be non-competitive or partial non-competitive versus both substrate (p-nitrophenol) and cofactor (PAPS). This inhibition mechanism indicates that meclofenamate, nimesulide and aspirin bind near enough to the substrate binding site to prevent catalysis but not affect dissociation of the substrate-enzyme complex. The inhibition of SULT1A1 by meclofenamate, nimesulide, salicylate and aspirin may be clinically relevant based on ratio of inhibition constant to predicted in vivo inhibitor concentration ([I]/IC(50) > 1).     

Sulfation of minoxidil by human liver phenol sulfotransferase

The N,O-sulfate of minoxidil (Mnx) is the active agent in producing the vasodilation and the hair-growth stimulating responses observed with Mnx treatment. In this report, Mnx sulfation activity was assayed in cytosol prepared from several normal human livers, and Mnx sulfation was shown to correlate significantly with the activity of the phenol-sulfating form of phenol sulfotransferase (P-PST) activity in the same livers. No correlation was observed between Mnx sulfation and the dopamine or dehydroepiandrosterone (DHEA) sulfotransferase activities present in human liver. Mnx sulfation also copurified with P-PST activity during the purification of P-PST from human liver. During the purification procedure, Mnx and p-nitrophenol sulfotransferase (P-PST) activities were resolved from the dopamine and DHEA sulfation activities catalyzed by the monoamine-sulfating form of phenol sulfotransferase (M-PST) and DHEA sulfotransferase respectively. Also, purified DHEA sulfotransferase was not capable of sulfating Mnx, and no data were obtained to indicate that Mnx is a substrate for M-PST. p-Nitrophenol, a substrate for P-PST, was demonstrated to be a competitive inhibitor of Mnx sulfation catalyzed by purified P-PST when Mnx was the variable substrate. These results indicate that Mnx is sulfated and, therefore, bioactivated by P-PST in human liver.     

非甾体抗炎药物临床应用分析

目的 通过评估非甾体抗炎药物在我院的使用情况,监测此类药物用药的合理性及药物利用情况.方法 分析2010年1-12月我院门诊处方.结果 共调查2010年度门诊处方304 089张,其中含NSMDs的处方30 503张,占总处方的10.03％.使用频率前10位的药物包括尼美舒利胶囊(12 278张)、25 mg阿司匹林(5098张)、美洛昔康分散片(3193张)、尼美舒利颗粒(2143张)、丙氧氨酚片(2139张)、萘丁美酮胶囊(2129张)、布洛芬片(968张)、尼美舒利分散片(906张)、洛索洛芬颗粒(676张)、布洛芬混悬液(506张).应用此类药物最多的是骨科(7354张,21.77％),其后依次为急诊科(6968张,20.62％)、神经内科(3879张,11.48％)、康复科(3536张,10.46％)、风湿科(2298张,6.80％)等.用药总量前10种NSAIDs的药物利用指数均＜1.结论 我院门诊非甾体抗炎药物的利用基本合理.     

采用报告比值比法挖掘非甾体抗炎药相关肝损伤信号

目的：探讨非甾体抗炎药（NSAID）相关肝损伤信号的挖掘及报告比值比（ROR）法在信号挖掘中的应用。方法以“肝”“、胆”为关键词检索国家药品不良反应（ ADR）监测系统2013年1月1日至12月31日接收到的北京市上报的ADR报告,将筛选后的报告中药物与肝损伤因果关系为“肯定”“、很可能”“、可能”的病例纳入肝损伤组,其他所有病例均纳入非肝损伤组。以NSAID为目标药物,其他所有药物为非目标药物,根据ROR计算公式计算NSAID相关肝损伤的ROR及其95%置信区间（ CI）,95%CI下限〉1提示出现ADR信号。结果经删重后共14657例患者纳入研究。肝损伤组626例,其中35例为NSAID相关肝损伤;非肝损伤组14031例。在35例NSAID相关肝损伤患者中,与NSAID单方制剂相关者30例,与复方制剂相关者5例,NSAID单、复方制剂及NSAID总体相关肝损伤ROR及其95%CI分别为1.78（1.22-2.61）、1.80（0.78-4.15）、1.76（1.24-2.51）;其中,NSAID单方制剂和总体相关肝损伤95%CI下限〉1,出现ADR信号。35例肝损伤共涉及37种药物,其中单方制剂帕瑞昔布、阿司匹林和复方制剂布洛伪麻、氨酚烷胺致肝损伤ROR及其95%CI分别为8.00（2.03-27.78）、2.45（1.43-4.21）、22.00（1.40-359.32）、和11.22（1.02-123.94）,均出现ADR信号。结论对NSAID相关肝损伤的风险应予关注。ROR法有助于挖掘ADR信号,为药物的安全使用提供预警。     

Inhibition of amyloidogenesis by nonsteroidal anti-inflammatory drugs and their hybrid nitrates

Poor blood-brain barrier penetration of nonsteroidal anti-inflammatory drugs (NSAIDs) has been blamed for the failure of the selective amyloid lowering agent (SALA) R-flurbiprofen in phase 3 clinical trials for Alzheimer's disease (AD). NO-donor NSAIDs (NO-NSAIDs) provide an alternative, gastric-sparing approach to NSAID SALAs, which may improve bioavailability. NSAID analogues were studied for anti-inflammatory activity and for SALA activity in N2a neuronal cells transfected with human amyloid precursor protein (APP). Flurbiprofen (1) analogues were obtained with enhanced anti-inflammatory and antiamyloidogenic properties compared to 1, however, esterification led to elevated Aβ(1-42) levels. Hybrid nitrate prodrugs possessed superior anti-inflammatory activity and reduced toxicity relative to the parent NSAIDs, including clinical candidate CHF5074. Although hybrid nitrates elevated Aβ(1-42) at higher concentration, SALA activity was observed at low concentrations (≤1 μM): both Aβ(1-42) and the ratio of Aβ(1-42)/Aβ(1-40) were lowered. This biphasic SALA activity was attributed to the intact nitrate drug. For several compounds, the selective modulation of amyloidogenesis was tested using an immunoprecipitation MALDI-TOF approach. These data support the development of NO-NSAIDs as an alternative approach toward a clinically useful SALA.     

我院门诊解热镇痛抗炎药物应用分析

目的分析门诊解热镇痛抗炎药物的使用情况,以期为临床合理选择解热镇痛抗炎药物提供参考。方法采用金额统计法,回顾性统计2016年6月门诊各类解热镇痛抗炎药物应用情况,对解热镇痛抗炎药物销售金额、构成比、各用药科室使用频率及其排序进行分析。结果 2016年6月门诊使用解热镇痛抗炎药物处方总金额为30 682.40元,占门诊药品销售总金额的1.17%;塞来昔布胶囊总销售金额排在解热镇痛抗炎药物第一位占47.33%;门诊解热镇痛抗炎药物使用频率最高的是外科门诊。结论解热镇痛抗炎药物合理使用疗效显著,不合理使用可导致严重不良反应。临床医师应正确合理地选择解热镇痛抗炎药物,更好地为患者的健康服务。     

非甾体抗炎药致老年人消化性溃疡的临床特点分析

目的:分析非甾体抗炎药(NSAIDs)所致的老年人消化性溃疡的临床特点。方法:回顾性调查上海市金山区亭林医院1999年1月至2006年12月间因消化性溃疡和(或)合并出血入院治疗的491例老年病人的临床资料,根据入院前1周内有无服用NSAIDs史将病人分为两组,对两组病人的临床资料进行分析比较。结果:服药组105例,未服药组386例。与未服药组比较,服药组病人的贫血更明显(P<0.05);复合溃疡和多发溃疡在服药组更多见(P<0.05)。服药组幽门螺杆菌的感染率为67.6%(71/105),未服药组为58.3%(225/386),两组间差别无统计学意义(P>0.05)。结论:应加强对老年人使用NSAIDs所致消化性溃疡的临床特点的认识,减少NSAIDs的不良反应。     

不同抗酸药防治非甾体类抗炎药胃粘膜损伤的疗效观察

目的探讨不同抗酸措施在防治服用非甾体类抗炎药患者的胃粘膜损伤中的作用。方法选择2004年1月-2006年11月我院门诊风湿性疾病患者120例,随机分为3组,均维持原来治疗方案,继续服用NSAIDs 3个月。同时,为防治胃粘膜损伤,A组应用奥美拉唑20 mg,口服,1次/d。B组应用雷尼替丁150 mg,2次/d,铝碳酸镁嚼片2 g,2次/d口服。C组应用雷尼替丁150 mg,2次/d。3组观察时间为3个月。观察胃粘膜损伤以及上消化道症状的发生率。结果A、B组较C组胃粘膜损伤的发生率以及上消化道症状发生率明显降低(P<0.05)。结论对服用非甾体类抗炎药患者,服用奥美拉唑或雷尼替丁+铝碳酸镁较服用雷尼替丁有更好的防治胃粘膜损伤的作用。     

Reactivity of nonsteroidal anti-inflammatory drugs with peroxidase: a classification of nonsteroidal anti-inflammatory drugs

Objectives To improve understanding of the essential effect of nonsteroidal anti-inflammatory drugs (NSAIDs) on prostaglandin H synthase (PGHS), the reactivity of NSAIDs with peroxidases and the tyrosyl radical derived from myoglobin was examined. Methods Horseradish peroxidase and myoglobin were used as models of peroxidase and cyclooxygenase of PGHS, respectively. Key findings From the results, a new classification of NSAIDs has been proposed. Class 1 includes the majority of NSAIDs, which reacted with horseradish peroxidase compound I, thus causing a spectral change by PGHS peroxidase and also including diminished electron spin resonance signals of the tyrosyl radical of myoglobin. They reduced compound I of horseradish peroxidase and scavenged the tyrosyl radical. The branched-chain mechanism by which the porphyrin radical is transferred to the tyrosine residue of the protein might be blocked by these NSAIDs. Class 2 includes salicylic acid derivatives that reacted only with the porphyrin radical and not with horseradish peroxidase compound II (oxoferryl species). Class 3 includes aspirin, nimesulide, tolmetin, and arylpropionic acid derivatives, including ibuprofen and the coxibs of celecoxib and rofecoxib, which are not substrates for horseradish peroxidase or PGHS peroxidase. Conclusions Understanding the essential mode of action of NSAIDs is particularly important for designing an effective therapeutic strategy against inflammatory diseases.     

Inhibitory potential of nonsteroidal anti-inflammatory drugs on UDP-glucuronosyltransferase 2B7 in human liver microsomes

Objective A number of nonsteroidal anti-inflammatory drugs (NSAIDs) are subject to glucuronidation in humans, and UDP-glucuronosyltransferase (UGT) 2B7 is involved in the glucuronidation of many NSAIDs. The objective of this study was to identify a NSAID with potent inhibitory potential against UGT2B7 using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Methods A rapid screening method for detecting the inhibitory potential of various drugs against UGT2B7 was established using a LC-MS/MS system. The effects of nine NSAIDs (acetaminophen, diclofenac, diflunisal, indomethacin, ketoprofen, mefenamic acid, naproxen, niflumic acid, and salicylic acid) against UGT2B7-catalyzed 3′-azido-3′-deoxythymidine glucuronidation (AZTG) were investigated in human liver microsomes (HLM) and recombinant human UGT2B7. Results Mefenamic acid inhibited AZTG most potently, with an IC₅₀ value of 0.3 μM, and its inhibition type was not competitive. The IC₅₀ values for diclofenac, diflunisal, indomethacin, ketoprofen, naproxen, and niflumic acid against AZTG were 6.8, 178, 51, 40, 23, and 83 μM, respectively, while those for acetaminophen and salicylic acid were >100 μM. The IC₅₀ values for NSAIDs against AZTG in recombinant human UGT2B7 were similar to those obtained in HLM. Conclusion The method established in this study is useful for identifying drugs with inhibitory potential against human UGT2B7. Among the nine NSAIDs investigated, mefenamic acid had the strongest inhibitory effect on UGT2B7-catalyzed AZTG in HLM. Thus, caution might be exercised when mefenamic acid is coadministered with drugs possessing UGT2B7 as a main elimination pathway.     

Sulfation of estrone and 17 beta-estradiol in human liver. Catalysis by thermostable phenol sulfotransferase and by dehydroepiandrosterone sulfotransferase.

Sulfation is a major pathway in humans for the biotransformation of estrogens. However, the nature of the enzymes that catalyze the sulfation of estrone (E1) and 17 beta-estradiol (E2) in human liver is unclear. Human liver contains at least three well-characterized cytoplasmic sulfotransferases, the thermostable (TS) and thermolabile (TL) forms of phenol sulfotransferase (PST) and dehydroepiandrosterone sulfotransferase (DHEA ST). Therefore, we determined optimal conditions for the assay of E1 and E2 ST activities in human hepatic cytosol to compare their properties and regulation with those of the three well-characterized human liver ST activities. Thermal inactivation studies showed that human liver E2 ST and TS PST had very similar thermal stabilities. The thermal inactivation profile of E1 ST suggested that this activity might be related to both DHEA ST and TS PST. Inhibition studies performed with 2,6-dichloro-4-nitrophenol (DCNP) also showed similar inhibition profiles for E2 ST and TS PST. Neither thermal inactivation nor DCNP inhibition studies indicated a possible relationship between TL PST activity and E1 or E2 sulfation. Experiments performed with 20 individual human liver samples showed highly significant correlations between activity levels of E2 ST and TS PST (rs = 0.944, p less than 0.0001), E1 ST and DHEA ST (rs = 0.845, p less than 0.0001), and, to a lesser degree, E1 ST and TS PST (rs = 0.608, p less than 0.01). Ion exchange chromatography of a human liver preparation, followed by assay of all five ST activities, confirmed the important roles played by TS PST and DHEA ST in the sulfation of E2 and E1. Similar results were found by study of the elution patterns of ST activities after ion exchange chromatography of human jejunal mucosal preparations. Partially purified TL PST, however, was unable to catalyze the sulfate conjugation of either E1 or E2. All of these results were compatible with the conclusion that, in human liver, TS PST is the enzyme predominantly responsible for the sulfate conjugation of E2, DHEA ST is the major enzyme responsible for the sulfation of E1, and TL PST does not appear to catalyze the sulfation of either E1 or E2.     

Case-control study on the association of upper gastrointestinal bleeding and nonsteroidal anti-inflammatory drugs in Japan

Objective Studies in Western populations have shown the association of nonsteroidal anti-inflammatory drugs (NSAIDs) and upper gastrointestinal bleeding (UGIB). The role of Helicobacter pylori infection in NSAIDs-related UGIB remains to be studied. We conducted a case-control study in Japan to investigate these related topics.Methods Cases of UGIB due to duodenal or gastric ulcer, or gastritis were identified in 14 study hospitals in various areas of Japan. For each case, two controls were identified from population registries in the same district. Information on drugs and other risk factors was obtained from 175 cases and 347 controls by telephone interviews. Anti-H. pylori antibody in the urine was measured in a single laboratory for all the cases and 225 controls.Results The odds ratio (OR) of UGIB was 5.5 for aspirin and 6.1 for other NSAIDs (NANSAIDs) (p<0.01). The OR for regular use was higher than for occasional use both for aspirin (7.7 vs 2.0) and NANSAIDs (7.3 vs 4.1). Loxoprofen (5.9), frequently used in Japan as a safe ‘prodrug’, was significantly associated with UGIB. The odds ratio for H. pylori infection was 4.9 and the relative excess risk due to the interaction between H. pylori and the use of NSAID was 1.2 (95% CI: −5.8–8.1).Conclusion NSAIDs including loxoprofen increase the risk of UGIB in Japan as in Western countries, with a similar magnitude of association. There was no evidence of biological interaction between NSAIDs and H. pylori infection.     

Inhibition of human thiopurine S-methyltransferase by various nonsteroidal anti-inflammatory drugs in vitro: a mechanism for possible drug interactions.

Thiopurine S-methyltransferase (TPMT) is a biotransformation phase II enzyme responsible for the metabolic inactivation of thiopurine drugs. The present study was carried out to investigate the inhibitory potential of 15 nonsteroidal anti-inflammatory drugs (NSAIDs) on human TPMT activity in vitro. TPMT activity was measured in pooled human erythrocytes in the absence and presence of various NSAIDs using the previously published high-performance liquid chromatography-UV method. To determine the inhibition type and K(i) value for each compound, we performed kinetic analysis at five different inhibitor concentrations close to the IC(50) value obtained in preliminary experiments. Naproxen (K(i) = 52 microM), mefenamic acid (K(i) = 39 microM), and tolfenamic acid (K(i) = 50 microM) inhibited TPMT activity in a noncompetitive manner. The estimated K(i) values for the inhibition of TPMT by ketoprofen (K(i) = 172 microM) and ibuprofen (K(i) = 1043 microM) indicated that the propionic acid derivatives were relatively weak inhibitors of TPMT. Our results suggest that coadministration of thiopurines and various NSAIDs may lead to drug interactions.     

Immunological characterization of human phenol sulfotransferase

The immunological characterization of the different forms of phenol sulfotransferase (PST) in a variety of human and nonhuman tissues is described. Immunoblotting techniques revealed that polyclonal antibodies raised to human platelet MII-PST reacted with polypeptides of 32 and 34 kDa from human platelet 100,000 x g supernatant solution. Immunoblot analysis of platelet 100,000 x g supernatant solution that was fractionated over a DEAE-cellulose column indicated a close correspondence of P-PST activity, as measured by phenol sulfation, and M-PST activity, as assessed by dopamine sulfation, with the 32 and 34 kDa polypeptides, respectively. Examination of various human tissues revealed the presence of immunologically detectable levels of P-PST in liver and adrenal gland whereas both M- and P-PST were detected in placenta at a 1/10,000 dilution of the antisera. Under these conditions, PST was undetectable in human frontal cortex, pituitary gland, kidney, lung, and jejunum. Further evaluation of human liver samples from four individuals indicated a strong correlation (r = 0.94) between the amount of 32-kDa immunoreactive protein and P-PST activity. Analysis of liver samples from several animal species (monkey, rat, mouse, guinea pig, and frog) revealed the presence of immunoreactive proteins of various molecular masses, suggesting that considerable homology may exist between human and nonhuman forms of PST.     

Immunomodulatory effect of nonsteroidal anti-inflammatory drugs (NSAIDs) at the clinically available doses

Non-steroidal anti-inflammatory drugs (NSAIDs) with cyclooxygenase (COX) inhibitory activity are commonly used in various inflammatory diseases. In this study, to examine the immunomodulatory effects of well known NSAIDs at clinically available doses, macrophage- and T cell-mediated immune responses such as tumor necrosis factor (TNF)-alpha release and nitric oxide (NO) production, cell-cell adhesion, phagocytic uptake and lymphocyte proliferation were investigated. NSAIDs tested significantly enhanced TNF-alpha release from lipopolysaccharide (LPS)-activated RAW264.7 cells at certain concentrations (fenoprofen, indomethacin, piroxicam, aceclofenac, diclofenac and sulindac) or in a dose-dependent manner (aspirin and phenylbutazone). Of NSAIDs, phenylbutazone and aspirin most potently attenuated NO production, although sulindac was the only compound with cytoprotective activity against LPS-induced cytotoxicity. Most NSAIDs used displayed weak or no modulatory effects on phagocytic uptake and CD29- or CD43-mediated cell-cell adhesion. Interestingly, however, phenylbutazone itself triggered cell-cell clustering under normal culture conditions and enhanced the phagocytic activity. Aspirin and phenylbutazone also dose-dependently attenuated CD4+ T cell proliferation stimulated by concanavalin A (Con A) and CD8+ CTLL-2 cell proliferation induced by interleukin (IL)-2. Sulindac only blocked CTLL-2 cell proliferation. These results suggest that NSAIDs may differentially exert immunomodulatory effects on activated macrophages and lymphocytes, and some of the effects may enforce NSAID's therapeutic effect against inflammatory symptoms.     

Renal adverse effects of nonsteroidal anti-inflammatory drugs

NSAIDs depress prostaglandins synthesis through inhibition of COX-1 that is involved in maintaining cell integrity and COX-2 that, although presents particularly in the kidneys, is overexpressed in response to inflammation. Both the beneficial and side effects of NSAIDs are, therefore, through their inhibition of COX enzymes. Introduction of COX-2-selective inhibitors has improved the safety profile of the drugs with regard to their most common side effect which occurs at the gastrointestinal level but has not rendered them less cardio-nephrotoxic. Renal side effects of NSAIDs are rare, sometimes transient and often reversible upon drug withdrawal. The incident rate and the severity of the renal side effect, however, increase in patients with risk factors such as those with diabetes, heart failure, renal dysfunction and in the elderly. The side effects range from electrolyte retention and reduce glomerular filtration to nephritic syndrome and chronic renal failure. These effects are shared among NSAIDs with evidence of dose and exposure dependency. There is no known predictor for the nephrotoxicity. However, a relationship has been found between high plasma concentration and the renal adverse effect of NSAIDs. The usefulness of therapeutic drug monitoring in patients with risk factors needs to be explored.     

Attenuation of epileptogenesis by nonsteroidal anti-inflammatory drugs in the rat

Subconvulsive doses (25 mg/kg) of pentylenetetrazol were administered at intervals of 4 days for 20 sessions, to induce kindling in conscious, free-moving rats, with chronically-implanted electrodes. This regimen induced an excitation of the CNS, which intensified over the 20 sessions. Periods of motor arrest, concurrent with bursts of electrocortical spike-wave activity, increased to clonic convulsions, concurrent with bursts of spike activity. Separate groups of rats were pretreated over the twenty sessions with nonsteroidal anti-inflammatory drugs (NSAIDs). Pretreatment with paracetamol produced a dose-related reduction in pentylenetetrazol-induced seizure activity. Pretreatment with 20 mg/kg mefenamic acid attenuated, while 60 mg/kg dose potentiated, the pentylenetetrazol-induced excitation. Pretreatment with 10 or 30 mg/kg ibuprofen had no significant effect, while 90 mg/kg was lethal, by itself, in 58% of the group. When all the groups received a single dose of pentylenetetrazol, three weeks after the twenty sessions, there were no significant differences between the groups in level of pentylenetetrazol-induced excitation, when compared to the control (saline-pretreated) group. This suggests that the effective NSAIDs had influenced the manifestation of, but not development of, epileptogenesis over the 20 sessions.