Non-steroidal Anti-inflammatory Acetylsalicylic Acid Research Articles

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.

Mechanochemical synthesis and characterization of a novel AAs–Flucytosine drug–drug cocrystal: A versatile model system for green approaches

Mechanochemistry approach is addressed herein for the green formation of a novel 1:1 drug–drug cocrystal (DDC) involving the antifungal prodrug Flucytosine (5FC) and the non-steroidal anti-inflammatory acetylsalicylic acid (AAS). This DDC was designed considering the huge market demand for new antifungal agents and the antifungal activity reported for some non–steroidal anti–inflammatory drugs, including AAS. The structure-properties characterization of the 5FC-AAS cocrystal was determined by X–ray diffraction (single crystal and powder), differential scanning calorimetry, thermogravimetric analysis, spectroscopic techniques (Raman and Fourier transform infrared), and hot–stage microscopy. The strong O–H…O and N–H…O hydrogen bonds formed between the hydroxyl group of AAS and the carbonyl and amine groups of two symmetry–related 5FC molecules maintain the crystalline packing, together with the same classical R22(8)homodimers observed in other 5FC cocrystals. The direct exposure under accelerated conditions (75% relative humidity/40 °C for 7 days) and at 12–months at room condition were also evaluated. Although the hygroscopicity addressed to 5FC was observed in the cocrystal during the stability studies, nevertheless this property discrepancy can easily be handled with the use of appropriate excipients and packing. DDC production was interestingly achieved by solvent evaporation, neat, and solvent-drop grinding (manually and in a ball-milling), resulting in a very fast production with suitable yield. In this sense, it is believed that this new model DDC will serve as a potential binary base cocrystal system for further developments of related higher dimensional cocrystals in this field, alongside other important drug molecules or in relation to the feasibility of using it therapeutically.

Innovative combination of QuEChERS extraction with on-line solid-phase extract purification and pre-concentration, followed by liquid chromatography-tandem mass spectrometry for the determination of non-steroidal anti-inflammatory drugs and their metabolites in sewage sludge

For the first time QuEChERS extraction of sewage sludge was combined with the automatic solid-phase pre-concentration and purification of the extract (following indicated as SPE) and LC-MS/MS analysis, for the determination of the non-steroidal anti-inflammatory drugs acetylsalicylic acid (ASA), diclofenac (DIC), fenbufen (FEN), flurbiprofen (FLU), ketoprofen (KET), ibuprofen (IBU) and naproxen (NAP), and their metabolites salicylic acid (SAL), 4′-hydroxydiclofenac (4′-HYDIC), 1-hydroxyibuprofen (1-HYIBU), 2-hydroxyibuprofen (2-HYIBU), 3-hydroxyibuprofen (3-HYIBU) and o-desmethylnaproxen (O-DMNAP). Various commercial pellicular stationary phases (i.e. silica gel functionalized with octadecyl, biphenyl, phenylhexyl and pentafluorophenyl groups) were preliminarily investigated for the resolution of target analytes and different sorbent phases (i.e. octyl or octadecyl functionalized silica gel and a polymeric phase functionalized with N-benzylpyrrolidone groups) were tested for the SPE phase. The optimized method involves the QuEChERS extraction of 1 g of freeze-dried sludge with 15 mL of water/acetonitrile 1/2 (v/v), the SPE of the extract with the N-benzylpyrrolidone polymeric phase and the water/acetonitrile gradient elution on the pentafluorophenyl stationary phase at room temperature. Matrix effect was always suppressive and in most cases low, being it ≤20% for ASA, DIC, FLU, KET, IBU, 1-HYIBU, 2-HYIBU, 3-HYIBU, NAP and O-DMNAP, and included in the range of 35–47% for the other analytes. Recoveries were evaluated at three spiking levels, evidencing almost quantitative values for HYIBUs and O-DMNAP; for ASA, SAL and KET the recoveries were included in between 50 and 76%, whereas for the other compounds they ranged from 36% to 55%. The proposed method showed better analytical performances than those so far published, being suitable for target compound determination in real samples from tens of pg g−1 to ng g−1 of freeze-dried sludge, with a total analysis time of 30 min per sample.

Synergistic and additive antiproliferative effects on human leukemia cell lines induced by combining acetylenehexacarbonyldicobalt complexes with the tyrosine kinase inhibitor imatinib

The tyrosine kinase inhibitor imatinib is successfully used in the treatment of chronic myeloid leukemia, but the occurrence of resistance phenomena can significantly limit therapeutic impact. Imatinib shows synergistic effects with cisplatin, suggesting that the coadministration of different cytostatics might reestablish the efficacy of treatment. We recently demonstrated that cobalt alkyne (or acetylenehexacarbonyldicobalt) complexes induce antiproliferative activity in human leukemia and lymphoma cells. The present study evaluates the effects of cobalt alkyne compounds containing propargylic acid esters on human acute (HL-60) and chronic myeloid (LAMA-84 and CML-T1) leukemia cell lines. The cell growth inhibitory activities (IC50 values of 9.5μM and higher) and induction of apoptosis (maximum 5.5-fold increase of single-stranded DNA at a drug concentration of 50μM) achieved with the single agents were moderate. Interestingly, suboptimal concentrations of the cobalt complexes (10μM) together with imatinib (0.1μM), when coadministered, showed an additive or synergistic effect on cellular proliferation inhibition. The most promising results were obtained with complexes containing ligands derived from the nonsteroidal antiinflammatory drugs acetylsalicylic acid and naproxene.

Multi‐residue liquid chromatography/tandem mass spectrometry method for the detection of non‐steroidal anti‐inflammatory drugs in bovine muscle: optimisation of ion trap parameters

A multi-residue liquid chromatography/tandem mass spectrometry method (LC/MS2) was developed for the detection of the non-steroidal anti-inflammatory drugs acetylsalicylic acid (via the marker residue salicylic acid), flunixin, phenylbutazone, tolfenamic acid, meloxicam and ketoprofen, in bovine muscle. After extraction of the bovine muscle with acetonitrile, the cleanup was performed using a Oasis HLB column. The evaporated eluate was reconstituted and analysed by LC/MS2. To obtain optimal detection of salicylic acid and phenylbutazone, the ion trap mass spectrometric parameters activation q and maximum ion injection time, respectively, were optimised. The activation q for salicylic acid was increased to obtain reliable detection of both salicylic acid and its product ion. The maximum ion injection time for the time segment containing phenylbutazone was decreased since there were not enough scans across the chromatographic peak of this compound. The multi-residue method was able to detect the different analytes below or at the maximum residue limit (MRL) or minimum required performance limit (MRPL) or, in the case of phenylbutazone and ketoprofen, at 100 and 20 microg kg(-1), respectively.

Yeast Eicosanoids II. The Influence of Non-Steroidal Anti-Inflammatory Drugs on the Life Cycle of Dipodascopsis

The effect of the non-steroidal anti-inflammatory drugs acetylsalicylic acid and indomethacin on the life cycles of Dipodascopsis tóthii and Dipodascopsis uninucleata is investigated. With ascosporogenesis as the most susceptible phase, it is found that these agents inhibit the completion of the life cycles of both species.

Inhibition of chloride/bicarbonate antiports in monkey kidney cells (vero) by non-steroidal anti-inflammatory drugs

Two chloride/bicarbonate antiport mechanisms are involved in the regulation of cytosolic pH (pH i) in Vero cells, namely Na +-dependent chloride/bicarbonate antiport to normalize pH i after acidification of the cytosol, and Na +- independent Cl −/HCO 3 − exchange to regulate pH i back to normal after alkalinization of the cytosol. We have tested the effects of the non-steroidal anti-inflammatory drugs acetylsalicylic acid (aspirin), salicylic acid, indomethacin and piroxicam on chloride/bicarbonate exchange and on chloride self exchange in Vero cells. All these drugs were found to inhibit both the Na +- independent and the Na +-linked chloride/bicarbonate antiport in a dose dependent manner. The Na +- independent chloride/bicarbonate antiport was inhibited by lower doses of the drugs than the Na +-linked antiport. The ability of the drugs to inhibit chloride self exchange did not vary much with varying external pH, indicating that the inhibitory effect is due to the anionic form of the drugs. Inhibition occurred immediately upon addition of the drugs, and it was rapidly reversible, indicating that the inhibitory effect is due to direct interaction of the drugs with chloride/bicarbonate antiport, and not to inhibition of prostaglandin synthesis. The relevance of our findings to the clinical effects of the drugs is discussed.

The effects of four platelet inhibitors on human platelets using two adhesiveness test systems.

The effects of the nonsteroidal anti-inflammatory drugs acetylsalicylic acid (ASA), naproxen (Naprosyn®, Syntex), and ibuprofen (Motrin®, Upjohn) and of the phosphodiesterase inhibitor VK 774 upon adhesion of human platelets to collagen fibers and to glass beads were compared after incubation with drug at two concentrations for 5 and 10 min. ASA significantly (p < 0.05−0.01) inhibited platelet adhesion to collagen fibers at the two concentrations (2.65 × 10−4 and 5.05 × 10−4 M) and after 5- and 10-min incubations. Naproxen inhibited adhesion significantly (p < 0.05−0.02) at the lower concentration (2.38 × 10−4 M) and at both incubation times. Ibuprofen did not significantly inhibit adhesion to collagen fibers. VK 774 significantly (p < 0.05−0.005) inhibited platelet adhesion to collagen fibers at both concentrations (2.38 × 10−4 and 4.54 × 10−4 M) and at both incubation times. ASA (1 × 10−3 M) produced slight but statistically significant (p < 0.05) inhibition of platelet adhesion to glass beads. Ibuprofen and naproxen (1 × 10−3 M) did not significantly inhibit adhesion in this test. VK 774 (1 × 10−3 M), however, significantly [Formula: see text] inhibited platelet adhesion to glass beads, to a greater extent than to collagen fibers. Naproxen (375 mg) was given orally in a single dose to five volunteer subjects. Bleeding time was doubled (p < 0.02) 2 h later and slightly but not significantly prolonged 24 h later. The second wave of platelet aggregation in response to ADP and adrenaline was completely blocked in all subjects 2 h after medication and still absent in one subject 24 h after medication, and higher concentrations of ADP and adrenaline were required to induce the second wave of aggregation in the remaining four subjects (4.2 × 10−6 M ADP and 5.6 × 10−5 M adrenaline in control period and 9.6 × 10−6 M ADP and 11.1 × 10−5 M adrenaline following drug administration). Collagen-induced aggregation was markedly inhibited 2 h after medication. These findings are in contrast with previously reported results with ibuprofen and suggest that naproxen induces greater impairment of hemostasis than does ibuprofen.