Properties Of Non-steroidal Anti-inflammatory Drugs Research Articles

Solvatochromism, preferential solvation and excited state dipole moments of flufenamic acid in different solvent polarities

The influence of pure solvent and binary solvent mixtures on the optical properties of non-steroidal anti-inflammatory drug (NSAID) molecule, specifically flufenamic acid (FLA), has been studied using absorption and fluorescence spectroscopic techniques. A bathochromic shift is observed in the emission wavelength of FLA with an increase in the solvent polarity of pure solvents, attributed to the highest stability of the excited state. The spectral properties are correlated with Lippert–Mataga, solvent polarity parameter, Kamlet, and Catalan solvent polarity parameters, suggesting that both general and specific solvent effects influence the spectral properties, with general solvent effects dominating over specific solvent effects. Preferential solvation studies have been carried out in tetrahydrofuran (THF) and water solvent mixture, revealing variations in the preference for solvation of FLA as a function of the mole fraction of water. Solvatochromic data are utilized to estimate the excited state dipole moment in pure solvents using different solvatochromic methods, revealing that the excited state dipole moment of FLA is higher than its ground state counterpart. The increase in dipole moment in the excited state compared to the ground state is explained based on intramolecular charge transfer (ICT), with elaboration and discussion on the possible resonance structure corresponding to ICT, inferring the more polar nature of the excited state compared to the ground state. The effect of solute polarizability on the excited state dipole moment and change in dipole moment is also discussed.

Quantum computations of non-steroidal anti-inflammatory drug molecules using Density Functional Theory

This paper addresses the electronic states and physicochemical properties of non-steroidal anti-inflammatory drug (NSAID) namely: Ibuprofen, Ketoprofen and Flufenamic acid. The molecular structures are quantum chemically treated and geometrically optimised using density functional theory (DFT). The computed geometrical parameters are compared with the experimental data. The Mulliken charge analysis is accomplished to understand the partial charge distribution in molecules. The molecular electrostatic potential analysis has been used to identify the appropriate location for an electrophilic and nucleophilic attacking site. Frontier molecular orbitals of the molecules are estimated and analysed. The HOMO-LUMO energy gaps have been used to estimate the different global reactivity descriptors of the selected molecules under investigation. The FT-IR spectra of molecules are recorded experimentally and are computed using DFT. The potential energy distribution (PED) assignment for the computed vibrations is carried out. The NBO analysis has been used to find the most intensive interaction in molecules based on the Fock-matrix sta3bilization energy. The NLO parameters have been computed for molecules to check their suitability towards NLO based applications. The UV-absorption spectra and ADME parameters for titled molecules are also simulated.

Development of an electrochemical cyclooxygenase biosensor to evaluate target–drug viability and interactions

In this research, we developed a biosensor to analyze the anti-inflammatory properties of non-steroidal anti-inflammatory drugs (NSAIDs) and potential candidate molecules. The chemical interaction between commercial NSAIDs and their target, the cyclooxygenase-2 enzyme (COX), was studied using electrochemical techniques and electronic paramagnetic resonance (EPR). The modified electrode, COX@GCE biosensor, presented two main reduction processes. We observed that the first one involves one electron and one proton related to tyrosyl radical reduction to tyrosine, and the second comprises a one-electron reduction from the Fe (IV) to Fe (III) species present in the enzyme’s active site, which in the presence of hydrogen peroxide results in a catalytical process. Based on these results, an electrochemical mechanism of the COX enzyme is proposed. Additionally, the quantitative inhibitory interaction mechanisms of six commercial NSAIDs were studied using this biosensor. The NSAIDs acetylsalicylic acid, dipyrone, and ibuprofen presented the higher COX inhibitory percentual, being, therefore, the most effective NSAIDs among the studied group. Salicylic acid also presented a significant inhibition capability in both electrochemical and EPR studies. The effect of some NSAIDs, e.g., dipyrone and acetaminophen, can be explained by the redox inhibition hypothesis and can be related to the direct interaction and inactivation of the iron present in the enzyme’s active site. Ibuprofen and naproxen presented irreversible COX inhibition. Despite that, the EPR shows that these drugs have relatively weak interactions with the COX active site, suggesting they should interact with the external structures resulting in the inhibition of its activity. Finally, the developed enzymatic sensor presented a high potential to study the anti-inflammatory properties of well-known drugs and can be a useful tool in the development of new NSAIDs, in a more efficient way, therefore presenting high applicability in drug development, pharmaceutical, and biomedical applications in both academia and industry.

Comparative effectiveness of early initiation of oral nonsteroidal anti-inflammatory drug and oral acetaminophen therapies on the time to knee replacement in patients with knee osteoarthritis in Japan

BackgroundAlthough disease-modifying properties of nonsteroidal anti-inflammatory drugs (NSAIDs) for osteoarthritis (OA) have been reported, the effects of NSAIDs on OA progression remain controversial. The purpose of this study was to investigate the effect of early initiation of oral NSAID therapy on the progression of knee OA.MethodsIn this retrospective cohort study, we extracted data of patients newly diagnosed with knee OA between November 2007 and October 2018 from a Japanese claims database. The primary outcome was the time to knee replacement (KR), and the secondary outcome was the time to composite event including joint lavage and debridement, osteotomy, or arthrodesis in addition to KR. Weighted Cox regression analysis with standardized mortality/morbidity ratio (SMR) weight was performed to compare the outcomes between patients prescribed oral NSAID (NSAID group) and those prescribed oral acetaminophen (APAP) (APAP group) early after a diagnosis of knee OA. Propensity scores were calculated using logistic regression conditioned on potential confounding factors, and SMR weights were calculated using the propensity scores.ResultsThe study population comprised 14,261 patients, who were divided into two groups as follows: 13,994 in the NSAID group and 267 in the APAP group. The mean ages of patients in the NSAID and APAP groups were 56.9 and 56.1 years, respectively. Furthermore, 62.01% and 68.16% patients in the NSAID and APAP groups, respectively, were female. The NSAID group had a reduced risk of KR compared with the APAP group in the analysis using SMR weighting (SMR-weighted hazard ratio, 0.19; 95% confidence interval, 0.05–0.78). While no statistically significant difference was found for the risk of composite event between the two groups (SMR-weighted hazard ratio, 0.56; 95% confidence interval, 0.16–1.91).ConclusionsThe risk of KR in the NSAID group was significantly lower than that in the APAP group after accounting for residual confounding using SMR weighting. This finding suggests that oral NSAID therapy early after the initial diagnosis is associated with a reduced risk of KR in patients with symptomatic knee OA.

Towards the Development of Dual Hypouricemic and Anti-inflammatory Candidates: Design, Synthesis, Stability Studies and Biological Evaluation of Some Mutual Ester Prodrugs of Febuxostat-NSAIDs

Treatment of gout involves two basic approaches: reducing the serum uric acid mainly by xanthine oxidase inhibitors (XOIs) and alleviating the intensity of the accompanying acute arthritic inflammation using non-steroidal anti-inflammatory drugs (NSAIDs). Febuxostat (FEB) is the first non-purine XOI approved for the treatment of hyperuricemia and gout. The present study aims at combining the hypouricemic effect of FEB and the anti‐inflammatory (AI) properties of NSAIDs in a single entity by adopting the “mutual prodrug” approach. Accordingly, a series of seven ester prodrugs comprising basically FEB together with different NSAIDs namely, diclofenac (4), ibuprofen (5), ketoprofen (6), indomethacin (7), naproxen (8), ketorolac (9) and etodolac (10) was synthesized. All the investigated seven prodrugs (4–10) were equipotent or even superior to their corresponding parent drugs in the hypouricemic and AI activities, together with a gastrointestinal (GI) safety profile. Among this series, the prodrug FEB-DIC (4) showed excellent dual in vivo hypouricemic and anti-inflammatory activity (43.60 % and 15.96 %, respectively) when compared to the parent drugs FEB and diclofenac (36.82 % and 12.10 %, respectively) and its physical mixture (37.28 % and 12.41 %, respectively). Investigation of the in vitro chemical stability and hydrolysis of the prodrug (4) in aqueous and biological samples using a developed HPLC method confirmed its stability in various pHs, whereas rapid hydrolysis to the parent drugs in liver homogenate and human plasma was proven. Finally, it is concluded that the mutual prodrug approach could be successfully used in drug design and development for overcoming undesirable difficulties without losing the desired activities of the parent drugs.

Comparative analysis of the effect of diclofenac sodium and etoricoxib on energy metabolism in rat liver in the acute general cooling model

When the ambient temperature decreases, physiological mechanisms that prevent heat loss are activated. However, under cold stress hypothermia develops, which significantly disrupts the functioning of the body and can be transformed into life-threatening. Preventive use of nonsteroidal anti-inflammatory drugs, especially diclofenac sodium and etoricoxib, has been found to reduce the severity of cold trauma. Given that their frigoprotective effect can accompany the impact on the synthesis of eicosanoids when exposed to low temperatures, it is advisable to study mechanisms for preventing hypothermia independent of cyclooxygenase, particularly the influence on energy metabolism. The aim of the study was to figure out the effect of diclofenac sodium and etoricoxib on the indicators of energy metabolism in the liver of rats after acute general cooling. Experiments were carried out on 28 sexually mature male rats, which were given diclofenac sodium (7 mg/kg), etoricoxib (5 mg/kg), or solvent 30 minutes before cold trauma intragastrically (in the intact control and pathology control groups). Acute hypothermia was caused by exposure of animals for 2 hours at a temperature of -18°C. Rectal temperature was measured before and after acute general cooling. The content of lactate, pyruvate and adenosine triphosphate in the liver was measured and the lactate/pyruvate ratio was calculated. Diclofenac sodium, unlike etoricoxib, was found to significantly reduce the severity of hypothermia. Both nonsteroidal anti-inflammatory drugs prevent energy metabolism disorders caused by exposure to cold, namely, reducing the concentration of lactic acid and the ratio of lactate/pyruvate, increasing the content of pyruvate and adenosine triphosphate in the liver of animals. Etoricoxib normalizes the content of energy metabolism intermediates to their levels in intact animals. Diclofenac sodium has a similar effect, the expression of which is inferior to the selective cyclooxygenase-2 inhibitor. Therefore, when administered prophylactically before acute general cooling, diclofenac sodium effectively prevents hypothermia in rats, surpassing etoricoxib. Etoricoxib completely prevents a decrease in the content of pyruvate and adenosine triphosphate, as well as the accumulation of lactic acid in the liver. Diclofenac sodium is inferior to etoricoxib in its effect on energy metabolism, which indicates other mechanisms of frigoprotective action of a non-selective cyclooxygenase inhibitor. The frigoprotective and energotropic properties of nonsteroidal anti-inflammatory drugs dissociate.

Does immunosuppressive property of non-steroidal anti-inflammatory drugs (NSAIDs) reduce COVID-19 vaccine-induced systemic side effects?

To help stop the coronavirus disease 2019 (COVID-19) pandemic, vaccines are currently the most critical tool. However, the COVID-19 mRNA vaccines frequently cause systemic side effects shortly after the injection, such as fever, headache and generalized fatigue. In our survey, after receiving the second dose of the COVID-19 vaccine, 80% developed fever, 62% headache and 69% generalized fatigue. Among people who required antipyretics, the average durations of fever and headache were significantly shorter in those who took non-steroidal anti-inflammatory drugs (NSAIDs), such as aspirin, loxoprofen and ibuprofen, than those who took acetaminophen. In our patch-clamp studies, NSAIDs effectively suppressed the delayed rectifier K+-channel (Kv1.3) currents in T-lymphocytes and thus exerted immunosuppressive effects. Because of this pharmacological property, the use of NSAIDs should be more effective in reducing the vaccine-induced systemic side effects that are caused primarily by the enhanced cellular immunity.

EVALUATION OF THE THERAPEUTIC EFFECT OF THE COMBINED USE OF CRYOPRESERVED PLACENTA EXTRACT AND DICLOFENAC SODIUM IN EXPERIMENTAL RHEUMATOID ARTHRITIS BY HEMATOLOGICAL PARAMETERS

Relevance. Cryopreserved placenta extract (CPE) increase the resistance of the mucous membrane of the gastrointestinal tract to the damaging effects of nonsteroidal anti-inflammatory drugs. Preventive administration of CPE can reduce the ulcerogenic effect of meloxicam, ibuprofen, diclofenac sodium (DS) and others. There is evidence of CPE's own anti-inflammatory activity, which can be successfully combined with the pharmacological properties of nonsteroidal anti-inflammatory drugs, while improving their safety profile.
 Objective: to characterize the therapeutic activity of the combined use of CPE and DS according to hematological parameters in the model of experimental rheumatoid arthritis (RA).
 Materials and methods. Studies were performed on 28 nonlinear laboratory rats. The rats were divided into 4 groups: I (n = 7) – intact rats; II (n = 7) – rats with experimental RA; ІІІ (n = 7) – rats with experimental RA, treated with DN; IV (n = 7) – rats with experimental RA, treated with DN and CPE. Adjuvant arthritis was modeled by subplantar administration of complete Freund's adjuvant. Treatment was performed from 14 to 28 days. CPE was administered on days 14, 17, 20, 23 and 26, and DS – daily. Blood tests were performed on day 28 of the experiment.
 Results. The combined use of CPE and DS is accompanied by a more pronounced leveling of inflammatory signs by hematological parameters – erythrocyte clotting rate decreased by 72.2% (p<0.001), and the number of leukocytes decreased by 54.81% (p<0.001) relative to rats with adjuvant arthritis without treatment. There was a leveling of signs of anemia of chronic inflammation – the level of hemoglobin and erythrocytes increased (p<0,001) by 17.6% and 36.8%, respectively, relative to rats with adjuvant arthritis without treatment.
 Conclusions. The combined use of CPE and DS is superior in therapeutic activity to monotherapy with this nonsteroidal anti-inflammatory drug of experimental rheumatoid arthritis.

Effect of choline chloride and urea based deep eutectic solvent on the physicochemical properties of salicylic acid and salicylamide at T = (288.15 to 313.15) K

Physicochemical properties of non-steroidal anti-inflammatory drugs (NSAIDs), salicylic acid and salicylamide, in choline chloride and urea based deep eutectic solvent (DES), reline, have been determined at temperature range from 288.15 to 313.15 K. In this work, physicochemical parameters such as apparent molar volume (Vϕ), apparent molar isentropic compressibility (Kϕ,s) and relative viscosities (ηr) are studied for the said NSAIDs in water and aqueous reline solutions. The results indicate that in all cases, the solute–solvent interactions increases with increasing concentration of DES as well as temperature. In addition, the values of viscosity B-coefficient are determined by analyzing Jones-Dole equation. Hepler’s constant (∂2V0ϕ/∂T2) and limiting apparent molar expansivity (E0ϕ) are deduced as well. Finally, the intermolecular interactions are evaluated by making plots among various parameters and discussed in terms of competing interactions present in the ternary systems.

A comparative study of the hepatotrophic properties of non-steroidal anti-inflammatory drugs

Topicality. Recently, more and more works on the hepatotoxicity of NSAIDs have appeared in the literature. Presumably, all NSAIDs have hepatotoxicity, but the degree of adverse effects on the liver in different drugs is variable. Aim. To study the effect of different generations of NSAIDs on the functional state of the liver of the experimental animals. Materials and methods. Diclofenac sodium, piroxicam, indomethacin, meloxicam and celecocosib were selected for the comparative study. Changes in the functional state of the liver of intact rats during subchronic administration of the drugs selected, as well as their impact on the course of the model hepatitis were determined. The state of the liver was determined by the following indicators: the mass coefficient of the liver, the activity of alanine aminotransferase, alkaline phosphatase, the content of total protein, urea, cholesterol in the blood serum, the level of TBA-active products, diene conjugates, reduced glutathione, catalase and glycogen in the liver homogenate. Results and discussion. It was found that diclofenac, piroxicam, indomethacin in the doses of ED50 by the antiexudative activity when used for 14 days adversely affected the liver of intact animals, as well as worsened the course of the model hepatitis, i.e. had a pronounced hepatotoxic effect. Meloxicam and celecoxib did not show a pronounced adverse effect in the carbon tetrachloride hepatitis, but contributed to the deterioration of the functional state of the liver of intact rats, i.e. had a moderate hepatotoxic effect. Conclusions. By the level of hepatotoxicity the drugs studied can be arranged as follows: diclofenac > indomethacin > piroxicam > meloxicam > celecoxib.

Inclusion Complexes of Non-Steroidal Anti-Inflammatory Drugs with Cyclodextrins: A Systematic Review

Non-steroidal anti-inflammatory drugs (NSAIDs) are one of the most widely used classes of medicines in the treatment of inflammation, fever, and pain. However, evidence has demonstrated that these drugs can induce significant toxicity. In the search for innovative strategies to overcome NSAID-related problems, the incorporation of drugs into cyclodextrins (CDs) has demonstrated promising results. This study aims to review the impact of cyclodextrin incorporation on the biopharmaceutical and pharmacological properties of non-steroidal anti-inflammatory drugs. A systematic search for papers published between 2010 and 2020 was carried out using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) protocol and the following search terms: “Complexation”; AND “Cyclodextrin”; AND “non-steroidal anti-inflammatory drug”. A total of 24 different NSAIDs, 12 types of CDs, and 60 distinct inclusion complexes were identified, with meloxicam and β-CD appearing in most studies. The results of the present review suggest that CDs are drug delivery systems capable of improving the pharmacological and biopharmaceutical properties of non-steroidal anti-inflammatory drugs.

Comparative Theoretical Study of Stability, Lypophilicity, Dipole Moments, Acidity and Spectroscopic Properties of Non-Steroidal Anti-Inflammatory Drugs: Ibuprofen, Ketoprofen and Flurbiprofen

A comparative theoretical study of some physicochemical properties of R and S enantiomers of ibuprofen, ketoprofen and flurbiprofen is undertaken in order to understand their reactivity. To do this, DFT and TD-DFT methods at the B3LYP/6-311G(d,p) level theory are used. The partition coefficient determined is 3.72 for ibuprofen, 2.81 for ketoprofen and 4.12. for flurbiprofen. That means that these NSAIDs are characterized by a high lipophilicity. The calculated Gibbs energies show that the R enantiomer is the most stable in the case of ibuprofen and the S enantiomer in the case of ketoprofen and flurbiprofen. The study of acidity shows that S enantiomer of ibuprofen and R enantiomers of ketoprofen and flurbiprofen are the most acidic enantiomers. TD-DFT calculations show that, the absorption maxima (λmax) of ibuprofen and flurbiprofen correspond to the HOMO→LUMO transition. For ketoprofen, these are associated with the transition HOMO→LUMO+1. These results elucidate the reactivity of the investigated NSAIDs and could help to establish a classification their efficacy.

The chemical and toxicological properties of nonsteroidal anti-inflammatory drugs and methods for their determination

The chemical and toxicological properties of nonsteroidal anti-inflammatory drugs and methods for their determination

Correlation analysis based on the hydropathy properties of non-steroidal anti-inflammatory drugs in solid-phase extraction (SPE) and reversed-phase high performance liquid chromatography (HPLC) with photodiode array detection and their applications to biological samples

In the present work we analyzed the hydrophobicity and hydrophilicity properties of several non-steroidal anti-inflammatory drugs (NSAIDs) by investigating the structural changes of the dynamic hydrogen bond network in order to predict the extraction recovery of NSAIDs from biological fluids set by solid phase extraction (SPE). This work allows investigating the relationship between theoretical descriptors and experimental data using a parameter free method with a strong correlation (Pearson correlation 0.95, p-value 0.0003).The identification and quantification of analytes in human plasma were carried out by high performance liquid chromatography coupled with photodiode array detection (HPLC-PDA) using a Kinetex Evo C18 (150 x 4.6 mm I.D) protected by a guard column and a mixture of acetonitrile and 10 mM phosphate buffer (pH 2.5) (50:50, v/v) as mobile phase at isocratic conditions. Accuracy (BIAS%) ranged within −2.33% and + 8.05% while precision (RSD%) was less than 5.73%.The mean extraction recovery of the carprofen (IS) was 84.1% and the recovery of NSAIDs from human plasma ranged between 81.9% to 86.6%. LODs and LOQs for all the investigated NSAIDs were 0.003 and 0.01 μg/mL, respectively. The method was validated according to the ICH guide line in the range 0.010–20.0 μg/mL.

Dual functional monomers modified magnetic adsorbent for the enrichment of non-steroidal anti-inflammatory drugs in water and urine samples

According to the molecular properties of non-steroidal anti-inflammatory drugs (NSADs), a new adsorbent for magnetic solid phase extraction (MSPE) was designed and synthesized. Triethyl-(4-vinylbenzyl)aminium chloride and 4-vinylbenzeneboronic acid were utilized as dual functional monomers to copolymerize with divinylbenzene on the surface of pre-modified Fe3O4 nanoparticles. The prepared magnetic adsorbent (Fe3O4@TCVA) was characterized by elemental analysis, Fourier transform infrared, scanning electron microscopy, transmission electron microscopy and vibrating sample magnetometer. Due to the abundant boronic acid, quaternary amine and phenyl groups, the Fe3O4@TCVA displayed satisfactory extraction performance for target NSADs (diclofenac acid, ibuprofen and mefenamic acid) by means of B-N coordination, anion-exchange, π-π and hydrophobic interactions. Under the optimized conditions, the Fe3O4@TCVA/MSPE was combined with high-performance liquid chromatography with diode array detection (HPLC-DAD) to sensitively analyze NSADs in water and human urine samples. Results indicated that the limits of detection for water and urine samples were in the ranges of 0.014–0.031 μg/L and 0.029–0.11 μg/L, respectively. The relative standard deviations for the intra-day and inter-day assay variability were below 10%. The applicability of the proposed Fe3O4@TCVA/MSPE-HPLC-DAD method was demonstrated by the successful extraction and quantification of trace levels of NSADs in real water and human urine samples. Satisfactory spiked recovery and reproducibility were achieved.

Cardiotropic action of combined use of celecoxib and amlodipine in rats sicked on adjuvant arthritis coupled with arterial hypertension

Drug therapy of rheumatoid arthritis combined with arterial hypertension is among actual medical objectives. The complexity of pharmacological treatment of comorbid state is due to not only pathological process severity, insufficient efficacy and side effects of disease modifying and symptomatic drugs but also property of nonsteroidal anti-inflammatory drugs (NSAIDs) to aggravate already existed arterial hypertension in patients with rheumatoid arthritis or to increase pressure. Many hypotensive drugs loose or don’t manifest their activity when used in combination with NSAIDs. High risk of cardio toxicity is registered for one of NSAIDs group – coxibs. The cardio safety of combined use of coxibs and hypotensive drugs on the ground of comorbid pathology is studied not enough.
 These aspects had predefined the aim of this study – to investigate cardiotropic effects of celecoxib when administered in combinations with amlodipine on the ground of experimental rheumatoid arthritis coupled with arterial hypertension.
 Arterial hypertension was modeled in rats by method of salt load. On the basis of arterial hypertension rheumatoid arthritis was caused by full Freund adjuvant injection. Arterial blood pressure and heart rate registered on sphygmomanometer.
 It was found that comorbid state is followed by arterial hypertension and tachyarrhythmia. Celecoxib does not facilitate hypertension enhancing but leads to increasing heart rate. Amlodipine manifests specific pharmacological activity as hypotensive drug. The results obtained predefined opportunity of combined use of celecoxib with amlodipine on the ground of rheumatoid arthritis coupled with arterial hypertension.

Abstract 3057: Potentiation of LPS-induced toxicity in human hepatoma HepG2 cells by aspirin and its protection by N-acetyl cysteine

Abstract Bacterial lipopolysaccharides (LPS) induce cytotoxicity in vitro and inflammation-induced toxic responses in vivo. Use of nonsteroidal anti-inflammatory drugs (NSAIDs), such as aspirin, has been reported to be beneficial in inflammation-associated diseases like cancer and diabetes. Their precise molecular mechanisms, however, are not clearly understood. Our previous studies on aspirin treated HepG2 cells strongly suggest cell cycle arrest and induction of apoptosis associated with mitochondrial dysfunction. In the present study, we have further demonstrated that HepG2 cells treated with LPS alone or in combination with aspirin induces subcellular toxic responses which are accompanied by increase in reactive oxygen species (ROS) production, oxidative stress, apoptosis and mitochondrial respiratory dysfunction. The LPS/Aspirin induced toxicity was attenuated by pretreatment of cells with N-acetyl cysteine (NAC). Oxidative stress and glutathione-dependent redox-homeostasis in the mitochondria were altered more severely compared to the extramitochondrial compartment of cells. Pre-treatment of HepG2 cells with NAC has demonstrated selective protection in redox homeostasis and mitochondrial dysfunction. Our results suggest that altered glutathione (GSH)-redox metabolism, oxidative stress and mitochondrial function in HepG2 cells play a critical role in LPS/aspirin-induced cytotoxicity. These results may help in better understanding the pharmacological and toxicological properties of NSAIDs in cancer cells exposed to bacterial endotoxins. (Supported by Sheikh Hamdan Medical Research Grant and Terry Fox Cancer Research Funds and a fund from CMHS Research Committee). Citation Format: Haider Raza, Annie John, Jasmin shafarin. Potentiation of LPS-induced toxicity in human hepatoma HepG2 cells by aspirin and its protection by N-acetyl cysteine. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3057. doi:10.1158/1538-7445.AM2015-3057

P7.05 - Metal complexes of Meloxicam and Isoxicam decrease viability and proliferation of virus-transformed cancer cells

ABSTRACT Introduction: There is growing evidence supporting the potential antitumor properties of non-steroidal anti-inflammatory drugs (NSAIDs). They are among the most consumed drugs over the world, including patients with cancer - a fact which further enhances the interest in the influence of these preparations on the survival and proliferative activity of tumor cells. It is assumed that for the realization of their biological activity is important linking to metal ions in the body. The aim of our study was to evaluate the effect of metal (CuII, ZnII, CoII, NiII) complexes of selective (Meloxicam) and non-selective (Isoxicam) NSAIDs on viability and proliferation of virus-transformed cancer cells. Materials and Methods: Permanent cell lines established from transplantable tumors in chicken (LSCC-SF-Mc29 –E7 - liver cancer, induced by the myelocytomatosis retrovirus Mc29) and rat (LSR-SF-SR–sarcoma, induced by RSV strain Schmidt-Rupin) were used as model systems. The investigations were performed by MTT test, neutral red uptake cytotoxicity assay, crystal violet staining, double-staining with acridine orange and propidium iodide, colony forming method and FACS analysis. The compounds were applied at a concentration range of 10-500 µg/ml for 24-120 h (short-term assays with monolayer cultures) and 18 days (long-term assay with 3D colonies). Results: The compounds examined decrease significantly viability and proliferation of treated cells as well as their ability to grow in semisolid medium in a time- and concentration-dependant manner. Cytopathological changes (including chromatin fragmentation and apoptotic bodies) and lower percentage of cells in G2 / M phase were also found. Conclusions: Cu(II) complexes with Meloxicam and Isoxicam are found to be the most active cytotoxic and cytostatic agents. They are highly effective against LSCC-SF-Mc29 cells that contain v-myc gene – disregulation of its cellular analogues is involved in pathogenesis of > 80% of human cancers. Applied independently, the ligands express lower cytotoxic/cytostatic activity as compared to metal complexes. Acknowledgements: Supported by Grant DFNI - Б 02/30 from 12.12.2014, Bulgaria, and a bilateral project between Bulgarian Academy of Sciences and Romanian Academy

Expression of pleiotrophin, an important regulator of cell migration, is inhibited in intestinal epithelial cells by treatment with non-steroidal anti-inflammatory drugs

Non-steroidal anti-inflammatory drugs (NSAIDs) are among the most widely used drugs for the suppression of inflammation and pain. However, the analgesic properties of NSAIDs are also associated with significant negative side effects, most notably in the gastrointestinal (GI) tract. Increasingly, evidence indicates that the ulcerogenic properties of some NSAIDs are not exclusively the result of inhibition of cyclooxygenase isoforms in the GI tract, and other mechanisms, including inhibition of cell migration and epithelial restitution, are being explored. Recently, microarray analysis was used to identify potential novel targets of NSAID activity in intestinal epithelial cells. Treated cells exhibited significant reductions in the gene expression of pleiotrophin (PTN), a cytokine and growth factor known to participate in angiogenesis and bone growth. This report aimed to confirm the microarray results reported previously, and to measure protein expression of PTN in intestinal epithelial cells. Furthermore, we also examined the effects of exogenous PTN on cell migration in the presence and absence of either NSAIDs with variable ulcerogenic potential or PTN-specific siRNA. Our results demonstrated that indomethacin and NS-398, two NSAIDs with ulcerogenic potential significantly decrease both gene and protein expressions of PTN in IEC-6 cells and protein expression in IEC-6-Cdx2 cells. Additionally, cell migration experiments with PTN siRNA showed that PTN is an important mediator of IEC-6 cell migration, and addition of exogenous PTN partially restores the deficits in cell migration caused by treatment with indomethacin and NS-398. Finally, measurement of PTN protein expression in the GI tract of horses treated with phenylbutazone showed that PTN expression is reduced by NSAIDs in vivo. Our results show that PTN is an important mediator of cell migration in IEC-6 cells, and PTN is a potential target through which NSAIDs may inhibit cell migration, epithelial restitution, and wound healing in the GI tract.

Abstract LB-172: MicroRNAs mediate the inhibition of tumor cell invasion by sulindac

Abstract A large body of evidence indicates that nonsteroidal anti-inflammatory drugs (NSAIDs) have strong antineoplastic properties against a broad range of cancer types in early or late stage animal models of tumorigenesis. However, their use in humans for chemoprevention is limited by toxicities resulting from cyclooxygenase (COX) inhibition and the suppression of physiologically important prostaglandins. The anticancer properties of NSAIDs are generally attributed to the suppression of tumor cell proliferation and induction of apoptosis by inhibiting cyclooxygenase (COX), although numerous studies suggest that a COX-independent mechanism may either contribute to or be fully responsible for their activity. The effect of NSAIDs on tumor cell motility and invasion has not been well studied, despite clinical studies showing that certain NSAIDs can significantly increase the survival of patients with metastatic disease. Our results show that the NSAID, sulindac sulfide (SS) can potently inhibit the motility of human breast (MDA-MB-231 and SUM1315) and colorectal (HCT116 and HT29) tumor cells by a mechanism that is distinct from its tumor cell growth and COX inhibitory activity. To study the molecular basis for this activity, we investigated the involvement of microRNA (miRNA). A total of 132 miRNAs were found to be altered in response to SS treatment including miR-10b, miR-17, miR-21, and miR-9, which have been previously implicated in tumor invasion and metastasis. We confirmed that these miRNAs can stimulate tumor cell invasion and show that SS can attenuate their invasive effects (individual and cooperative) by down-regulating their expression. Employing luciferase and chromatin immunoprecipitation (ChIP) assays, NF-κB was found to bind the promoters of all four miRNAs to regulate their expression at the transcriptional level. Results from the immunofluorescence and Western Blotting assays show that SS can inhibit the translocation of NF-κB to the nucleus by decreasing the phosphorylation of IKKβ and IκB, which leads to down-regulation of these miRNAs. Analysis of the promoter sequences of the miRNAs suppressed by SS revealed that 81 of 115 sequences contained NF-κB binding sites. Our results demonstrate that SS can inhibit tumor cell invasion by suppressing NF-κB mediated transcription of miRNAs, which illustrate the mechanistic basis of this novel anticancer property of sulindac on tumor progression and metastasis. We expect these results are capable of supporting future clinical trials of sulindac in patients with metastatic disease. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-172. doi:1538-7445.AM2012-LB-172