Anti-inflammatory Nimesulide Research Articles

Insights into the effect of glucose on the binding between human serum albumin and the nonsteroidal anti-inflammatory drug nimesulide

Glucose interacts with human serum albumin (HSA, the main protein responsible for the biodistribution of drugs in the bloodstream) and consequently affects the binding capacity of exogenous compounds. Thus, in this work, the interactive profile between HSA and the anti-inflammatory drug nimesulide (NMD, used mainly by patients with diabetic neuropathy to relieve acute or chronic pains) was characterized in nonglycemic, normoglycemic (80 mg/dL), and hyperglycemic (320 mg/dL) conditions by biophysics techniques. There is a spontaneous and ground-state association HSA:NMD under physiological conditions. Therefore, the Stern-Volmer constant (Ksv) can also be used to estimate the binding affinity. The Ksv values for nonglycemic, normoglycemic, and hyperglycemic conditions are around 104 M−1, indicating a moderate affinity of NMD to albumin that was slightly improved by glucose levels. Additionally, the binding is enthalpically and entropically driven mainly into subdomains IIA or IIIA. The binding perturbs weakly the α-helix content of albumin, however, glucose potentially stabilizes the tertiary structure, decreasing the structural perturbation upon NMD binding and improves the complex HSA:NMD stability. Overall, the biophysical characterization indicated that glucose levels might slightly positively impact the pharmacokinetic profile of NMD, allowing NMD to achieve its therapeutical potential without affecting drastically its effective dosages.

Crystal structure of the tetra-ethyl-ammonium salt of the non-steroidal anti-inflammatory drug nimesulide (polymorph II).

The crystal structure of the tetra-ethyl-ammonium salt of the non-steroidal anti-inflammatory drug nimesulide (polymorph II) (systematic name: tetra-ethyl-ammonium N-methane-sulfonyl-4-nitro-2-phen-oxy-anilinide), C8H20N+·C13H11N2O5S-, was determined using single-crystal X-ray diffraction. The title compound crystallizes in the monoclinic space group P21/c with one tetra-ethyl-ammonium cation and one nimesulide anion in the asymmetric unit. In the crystal, the ions are linked by C-H⋯N and C-H⋯O hydrogen bonds and C-H⋯π inter-actions. There are differences in the geometry of both the nimesulide anion and the tetra-ethyl-ammonium cation in polymorphs I [Rybczyńska & Sikorski (2023 ▸). Sci. Rep. 13, 17268] and II of the title compound.

Structural insight and in silico prediction of the pharmacokinetic parameters and toxicity of alkaline earth metal compounds strontium and barium with the non-steroidal anti-inflammatory drug nimesulide.

In the crystals of alkaline earth metal compounds strontium and barium with the non-steroidal anti-inflammatory drug nimesulide, the strontium cation is nine-coordinated with a distorted tricapped trigonal prismatic geometry TCTPR-9, whereas the ten-coordinated barium ion exhibits a distorted tetracapped trigonal prismatic geometry TCTPR-10.

Interaction of anti-inflammatory drug nimesulide with ionic and non-ionic surfactant micelles: Insights from spectral and bioinformatics approach

The present work reports the interaction of anti-inflammatory drug nimesulide with cationic (CTAB, HDPC), anionic (SDS) and non-ionic (Triton X-100) surfactant micelles. The drug-micelles systems were studied by UV–Vis absorption spectroscopy to estimate the binding constant (Kb), partition coefficient (Kx) and the free energy of binding and partition (ΔGb, ΔGx). The values of Kb indicate a stronger interaction between nimesulide and cationic micelles due to cumulative electrostatic and hydrophobic forces. Also, the higher values of Kx obtained for HDPC and CTAB micelles point out a higher drug concentration in cationic micelles than in the surrounding aqueous medium, as against SDS and Triton X-100 micelles. The bioinformatic methods were used to evaluate the pharmacokinetic, pharmacogenomic and pharmacodynamic profiles of the nimesulide, CTAB, HDPC, SDS, and Triton X-100. Our results showed that all compounds exhibit good intestinal absorption, good BBB and CNS permeabilities, and low toxicity features. Also, the ecotoxicity of compounds was quantified. Various pharmacogenomic features of compounds were recorded. Supplementary, we conducted an extensive analysis based on various aspects of the structural and functional features of compounds, such as their electronic (dipole moment, frontiers orbital energies), steric, and hydrophobic features. Experimental and theoretical results show that CATB and HDPC cationic micelles may be better drug delivery agents for nimesulide.

Вплив німесуліду та нового похідного 4-тіазолідинону на гематологічні параметри в умовах експериментального запального процесу

The aim of the work was to identify the anti-inflammatory effect of a newly synthesized drug from the group of 4-thiazolidinones. Thus, a comparative study of the effect of two agents was conducted: the drug Les6490 and the non-steroidal anti-inflammatory drug nimesulide. This drug belongs to the group of non-steroidal anti-inflammatory drugs (NSAIDs), which have pronounced anti-inflammatory, analgesic and antipyretic effects. Nimesulide belongs to highly selective inhibitors of cyclooxygenase and its isoenzymes (COX-2) and is used for treatment in various pathologies of the musculoskeletal system. The newly synthesized compound Les6490 belongs to the group of 4-thiazolidinone derivatives. The thiazolidinone ring is part of many existing potential antibacterial and anti-inflammatory agents, and can be combined with pyrazole fragments, which are pharmacophores of the same structure, and can contribute to increased therapeutic efficacy. The study of the peculiarities of hematological changes in rats during the experimental reproduction of the associated inflammatory process using the Freund’s adjuvant model has been conducted. As a result of the study of anti-inflammatory activity based on hematological indicators of the drugs — a new synthesized derivative of 4-thiazolidinones and a nonsteroidal anti-inflammatory drug nimesulide — their anti-inflammatory activity was revealed, which can be considered comparable to the slight advantage of the drug Les6490. At the same time, without affecting the quantitative level of erythrocytes with isolated administration, under the conditions of the inflammatory process, nimesulide and Les6490 caused a significant increase in the total number of erythrocytes. No effect on the amount of erythrocytes was found. The analysis of leukocyte formula suggests the evidence that the activity of Les6490 from the group of 4-thiazolidinones is to some extent more pronounced than the activity of nimesulide. The most pronounced changes in the leukocyte formula during AF-induced inflammation were observed on the part of neutrophils: the effect of the studied substance Les6490 was more pronounced compared to the effect of nimesulide. The introduction of the new compound Les6490 partially caused the normalization of monocytes, and under the conditions of the AF-associated inflammatory process, no significant changes were observed.

Molecular Characterization of a First-in-Human Clinical Response to Nimesulide in Acute Myeloid Leukemia.

Acute myeloid leukemia (AML) is a hematologic malignancy associated with high morbidity and mortality. Here we describe a case of a patient with AML who presented a partial response after utilization of the non-steroidal anti-inflammatory drug nimesulide. The response was characterized by complete clearance of peripheral blood blasts and an 82% decrease of bone marrow blasts associated with myeloblast differentiation. We have then shown that nimesulide induces in vitro cell death and cell cycle arrest in all AML cell lines (HL-60, THP-1, OCI-AML2, and OCI-AML3). Weighted Correlation Network Analysis (WGCNA) of serial whole-transcriptome data of cell lines treated with nimesulide revealed that the sets of genes upregulated after treatment with nimesulide were enriched for genes associated with autophagy and apoptosis, and on the other hand, the sets of downregulated genes were associated with cell cycle and RNA splicing. Serial transcriptome of bone marrow patient sample confirmed the upregulation of genes associated with autophagy after the response to nimesulide. Lastly, we demonstrated that nimesulide potentiates the cytotoxic in vitro effect of several Food and Drug Administration (FDA)-approved chemotherapy drugs used in AML, including cytarabine.

Adenoprosin in the complex treatment of lower urinary tract symptoms in combination with prostatitis of category III

Introduction.The problem of effective and safe therapy of patients with benign prostatic hyperplasia and prostatitis of category III remains relevant, despite the numerosity of the drugs used in different groups. Under these conditions, the search for new pharmacological agents continues.The study objectivewas to evaluate the efficacy of Adenoprosin in the treatment of lower urinary tract symptoms and erectile dysfunction in patients with chronic pelvic pain syndrome and prostate hyperplasia.Materials and methods.The 90-day study included 60 patients aged 40 to 60 years with chronic pelvic pain syndrome and erectile dysfunction on the background of benign prostatic hyperplasia. Patients of the main group received alfa-1-adrenoreceptor blocker tamsulosin for 30 days in combination with the nonsteroidal anti-inflammatory drug nimesulide for 5 days and Adenoprosin suppositories. The men of the control group took tamsulosin for 30 days and nimesulide for 5 days. Over the next 60 days, the dynamics was evaluated based on the data from the NIH-CPSI (National Institutes Health Chronic Prostatitis Symptom Index), IPSS (International Prostate Symptom Score), QoL (Quality of Life Scale), AMS (Aging Male Symptoms), IIEF-5 (International Index of Erectile Function); urination diary analysis; general urine analysis; blood test for the level of prostate specific antigen, glucose; prostate secretion analysis; transrectal ultrasound examination of the prostate gland with measurement of residual urine; uroflowmetry.Results.Urinary frequency decreased at day 30 in the treatment group from 12.7 ± 1.4 to 7.3 ± 1.8 micturitions per day, in the control group from 13.1 ± 1.8 to 9.6 ± 1.1; subsequently (on days 60 and 90) positive effect persisted in the treatment group, while in the control group urinary frequency increased. Per the NIH-CPSI questionnaire, symptoms decreased at day 30: in the treatment group from 25.4 ± 2.1 to 12.3 ± 1.9 points, in the control group from 24.3 ± 1.7 to 17.6 ± 0.4 points; in the treatment group the trend towards symptom decrease remained, while in the control group symptoms increased at day 90. IPSS score at day 30 decreased in the treatment group from 12.9 ± 0.5 to 5.2 ± 1.4, in the control group from 14.1 ± 0.7 to 8.2 ± 0.9; further decrease was observed in both groups at day 60, but at day 90 the score increased (higher in the control group). Quality of life (QoL) improved in both groups at day 30 (in the treatment group 1.4 ± 0.6 points, in the control group 3.1 ± 0.8). Erectile function improved per the IIEF-5 questionnaire: in the treatment group from 11.5 ± 1.1 to 18.9 ± 1.4 points at day 30 with further positive tendency, in the control group from 10.7 ± 1.4 to 15.3 ± 1.1 points with subsequent negative dynamics. Uroflowmetry showed positive dynamics of Qmaxat the end of treatment in the treatment (from 8.3 ± 1.5 to 14.7 ± 1.1 mL/s) and control (from 8.8 ± 1.3 to 13.1 ± 1.6 mL/s) groups, with its further decrease. Prostate volume in the treatment group was about 33.5 ± 3.1 cm3at day 90 (with baseline volume 42.6 ± 2.5 cm3), in the control group it was 41.6 ± 1.9 cm3(with baseline volume of 43.1±1.4 cm3). No significant changes in urine tests, prostatic fluid, prostate-specific antigen level, and blood glucose were observed during the treatment.Conclusions.The study shows that administration of tamsulosin with Adenoprosin in patients with BPH and category III prostatitis is advantageous compared to administration of only alfa-1-adrenoreceptor blockers.

Fast crystal growth of amorphous nimesulide: implication of surface effects.

Understanding crystallization behaviors is of utmost importance for developing robust amorphous pharmaceutical solids. Herein, the crystal growth behaviors of amorphous anti-inflammatory drug nimesulide (NIME) are systemically investigated in the glassy and supercooled liquid state as a function of temperature. A sudden over-tenfold increase is observed in the bulk crystal growth of NIME on cooling below its glass transition temperature (Tg). This fast growth behavior is known as a glass-to-crystal (GC) mode and has been reported in some molecular glasses. Fast surface crystal growth of NIME can persist up to Tg + 57°C with a weak jump in its growth rates at 30-40°C. In addition, surface crystal growth and GC growth of NIME exhibit an almost identical temperature dependence, supporting the view that GC growth is indeed a surface-facilitated process. Moreover, the bubble-induced fast crystal growth of NIME is observed in the interior of its supercooled liquid with approximately the same growth kinetics as surface crystal growth. These findings are relevant for a full understanding of the surface-related crystallization behaviors and physical stability of amorphous pharmaceutical formulations.

Drug repurposing for Chagas disease: In vitro assessment of nimesulide against Trypanosoma cruzi and insights on its mechanisms of action.

Chagas disease is a neglected illness caused by Trypanosoma cruzi and its treatment is done only with two drugs, nifurtimox and benznidazole. However, both drugs are ineffective in the chronic phase, in addition to causing serious side effects. This context of therapeutic limitation justifies the continuous research for alternative drugs. Here, we study the in vitro trypanocidal effects of the non-steroidal anti-inflammatory drug nimesulide, a molecule that has in its chemical structure a toxicophoric nitroaromatic group (NO2). The set of results obtained in this work highlights the potential for repurposing nimesulide in the treatment of this disease that affects millions of people around the world.

The non-steroidal anti-inflammatory drug nimesulide kills Gyps vultures at concentrations found in the muscle of treated cattle

Throughout South Asia, cattle are regularly treated with non-steroidal anti-inflammatory drugs (NSAIDs) and their carcasses are left for scavengers to consume. Residues of the NSAID diclofenac in cattle carcasses caused widespread mortality and catastrophic population declines in three species of Gyps vulture during the 1990s and 2000s. Diclofenac is now banned, but other NSAIDs are used in its place. Different lines of evidence, including safety testing in Gyps vultures, have shown that some of these other NSAIDs are toxic, or probably toxic, to vultures. The NSAID nimesulide is widely available and commonly used, and has been found in dead vultures with signs of renal failure (i.e. visceral gout) and without the presence of diclofenac and/or other vulture-toxic NSAIDs. Nimesulide is therefore probably toxic to vultures. Here, we report safety testing of nimesulide in Gyps vultures. In a controlled toxicity experiment, we gave two vultures the maximum likely exposure (MLE) of nimesulide calculated from initial pharmacokinetic and residue experiments in cattle. Two other control birds were given an oral dose of water. Both vultures dosed with nimesulide died within 30 h, after showing outward signs of toxicity and increases in biochemical indicators of renal failure. Post-mortem examinations found extensive visceral gout in both vultures. Both control vultures survived without biochemical indicators of renal failure. With this evidence, we call for an immediate and comprehensive ban of nimesulide throughout South Asia to ensure the survival of the region's Critically Endangered vultures. More generally, testing the impacts of drugs on non-target species should be the responsibility of the pharmaceutical industry, before their veterinary use is licensed.

Extractive spectrophotometric determination of anti-inflammatory drug nimesulide in pharmaceutical formulations and human plasma

The goal of the present work is to develop simple, rapid, and sensitive extractive spectrophotometric methods for the assay of nimesulide in pharmaceutical formulations and human plasma. The methods depend on the formation of colored ion-pair complexes between nimesulide and five different reagents, bromocresol green (BCG), bromocresol purple (BCP), bromothymol blue (BTB), brilliant blue G (BBG) and methyl orange (MO) in Britton-Robinson buffer solutions of pH 2.0-3.5. The colored products were extracted in chloroform and absorbance measured at 412, 410, 407, 502 and 482 nm, respectively. The analytical parameters and their effects on the reported systems were investigated. The extracts are intensely colored and very stable at room temperature. The calibration graphs were linear over the concentration range of 2-18 μg/mL for BCG, BTB and BBG, 2-16 μg/mL for BCP and 2-14 μg/mL for MO. The stoichiometry of the reaction was found to be 1 : 1 in all cases and the conditional stability constant (K f) of the complexes were calculated. The proposed methods were successfully extended to pharmaceutical preparations in tablet form. Excipients used as additives in commercial formulations did not interfere with the analysis. The proposed methods can be recommended for quality control and routine analysis where time, cost effectiveness and high specificity of the analytical technique are of great importance.

Photolysis and photocatalysis of the non-steroidal anti-inflammatory drug Nimesulide under simulated solar irradiation: Kinetic studies, transformation products and toxicity assessment

In this study, the degradation of Nimesulide (NIM), a non-steroidal anti-inflammatory drug, using photolysis, heterogeneous (TiO2 in dispersion) and homogeneous (photo-Fenton reactant) photocatalysis, under simulated solar light (SSL) radiation, was investigated. Various parameters affecting the degradation rate of the target compound during the applied processes were optimized. The efficiency of all treatments used (direct photolysis; TiΟ2/SSL; TiΟ2/Η2Ο2/SSL; TiΟ2/S2Ο82−/SSL; Fe3+/H2O2/SSL; Fe3+/S2O82−/SSL and [Fe(C2O4)3]3−/H2O2/SSL) was evaluated by means of initial reaction rate and mineralization. Moreover, the generated transformation products (TPs) by each basic process (photolysis; TiΟ2/SSL and Fe3+/H2O2/SSL) were identified, using liquid chromatography coupled to high resolution mass spectrometry, and their formation kinetic profiles were given. The main transformation routes of NIM were hydroxylation and fragmentation, for all three treatments applied. Finally, toxicity measurements were conducted using Microtox bioassay in order to evaluate the potential risk of NIM and its TPs to aqueous organisms. Although, the acute toxicity increased during the first stages of treatment the final outcome lead to very low toxicity levels even within 60 min of TiO2/SSL treatment. Concluding, the obtained results suggest that the photocatalytic degradation of NIM can lead to its complete elimination and simultaneously to the detoxification of the solution.

Nimesulide effects on the blood pro-oxidant–antioxidant status in lipopolysaccharide-challenged mice

The maintenance of the pro-oxidant–antioxidant equilibrium between ROS production and antioxidant protection systems is an important element of systemic defence and requires efficient control. The aim of the study was to monitor the dynamics of antioxidants and lipid peroxidation in mice challenged intraperitoneally with Escherichia coli (O111:B4) lipopolysaccharide (LPS) and to evaluate the antioxidant potential of the non-steroidal anti-inflammatory drug nimesulide. Albino mice were divided into three groups (n = 36). Group I received a single intraperitoneal (i.p.) injection with 25 μg/0.5 mL LPS. Thirty minutes before LPS, group II received orally (p.o.) 100 mg/kg nimesulide. The preparation was administered for 4 days. Group III received only nimesulide at the indicated dose for 4 days. The blood parameters were analysed at hour 0 (prior to treatment applied to each group), post treatment hours 6 and 24, and days 3, 5 and 9. Assayed parameters included catalase, reduced glutathione, albumin, glucose, ferric reducing ability of plasma (FRAP), malondialdehyde and oxidative stress index. LPS induced continuous hypoglycaemia, decreased catalase activity and reduced glutathione, but FRAP and albumin were preserved. The application of nimesulide alone did not alter oxidative stress index and enhanced FRAP. Its co-administration with LPS normalised reduced glutathione, decreased catalase and increased malondialdehyde concentrations and oxidative stress index. The application of nimesulide as antioxidant requires objective evaluation of associated benefits and risks.

The nitro-reduced metabolite of nimesulide: Crystal structure, spectroscopic characterization, ESI-QTOF mass spectrometric analysis and antibacterial evaluation

Here we present a synthetic procedure, spectroscopic characterization and single-crystal X-ray structure for the nitro-reduced metabolite of the anti-inflammatory drug nimesulide, hereby referred to as NMS-NH2. The nitro-reduced metabolite was synthesized using the Béchamp reduction (iron powder under acidic media), leading to the conversion of the nitrobenzene group of nimesulide to an aniline. Mass spectrometry, infrared and nuclear magnetic resonance spectroscopies data are also provided for NMS-NH2, and discussed in comparison to nimesulide. NMS-NH2 was also evaluated in terms of its antibacterial activities, considering that the free NH2 group could allow the compound to act as a dihydropteroate synthase inhibitor. NMS-NH2 had a modest antibacterial activity against P. aeruginosa (5.0 mg mL−1), which was not observed for NMS.

SPECTROPHOTOMETRIC EVALUATION OF NIMESULIDE IMPURITY D: 4-NITRO-2- PHENOXYANILINE USING 8-HYDROXYQUINOLINE AS OXIDATIVE COUPLING REAGENT

Objective: Spectrophotometric investigation of 4-nitro-2-phenoxyaniline (4N2PA), a probable intermediate during the synthesis of anti-inflammatory drug nimesulide, both in pure form and in the presence of nimesulide has been attempted. Methods: 4N2PA on diazotization followed by coupling with 8-hydroxyquinoline produces a crimson-colored complex which changes to deep violet on diluting with ethanol. Results: The complex showed maximum absorbance at 560 nm when evaluated spectrophotometrically with a detection limit of 0.005 μg/ml and quantification range of 0.05-3.0 μg/ml. The method has been statistically evaluated with respect to the International Council for Harmonisation guidelines and found to be accurate and precise. Conclusion: Pure tablet formulations of nimesulide do not respond to the method; however, the presence of minute amounts of 4N2PA in the drug as added impurity could be spectrophotometrically analyzed. Hence, the authors suggest that the reported technique could be a marker test for detecting the presence of 4N2PA in nimesulide formulations.

Adsorption of anti-inflammatory nimesulide by graphene materials: a combined theoretical and experimental study.

Interactions of anti-inflammatory nimesulide (NM) with different graphene material species were explored employing both ab initio calculations, based on Density Functional Theory (DFT), and a batch adsorption process. The adsorption of NM onto graphene, with and without a vacancy, reduced graphene oxide (rGO) and functionalized graphene nanoribbons was simulated, providing a good understanding of the adsorption process of the NM molecule onto graphene material surfaces. The theoretical results indicate a physisorption interaction between NM and all of the evaluated adsorbents. Based on batch adsorption experiments, it was found that rGO, obtained via a modified Hummers method, is a good nanoadsorbent for the removal of the anti-inflammatory NM from aqueous solutions. The general-order kinetic equation displays the best fit to the experimental data compared with pseudo-first order and pseudo-second order kinetics. The equilibrium data fitted well into the Liu isotherm equation, and the maximum sorption capacity for the adsorption of NM by rGO was 82.4 mg g-1 at 25 °C. Our results of the first principle calculations and the batch adsorption experiments point out that graphene materials are promising nanomaterials for extracting NM from aqueous solutions.

NIMESULIDE IN TREATMENT OF ACUTE AND CHRONIC PAIN IN CLINICAL PRACTICE

The most common reason for a patient to contact a doctor is the pain accompanying about 70% of all known diseases. The article introduces the basic principles for treating acute and chronic pain. The role of inflammation in pathogenesis of acute and chronic pain is shown. It provides information on the effectiveness and safety of a non-steroid anti-inflammatory drug (NSAIDs) Nimesulide in the treatment of acute and chronic pain. Information is provided on the variety of Nimesulide mechanisms that allow it to be used in various rheumatic diseases as well as in other areas of medicine. The results of clinical research and analyses are presented that confirm the high efficiency and good tolerability of Nimesulide. The years of experience in the application of the granular form of Nimesulide (Nimesil®) by domestic clinicians indicate a high rate of development and a expressed anagesic, anti-inflammatory effects combined with good tolerability, which makes it possible to use the drug successfully in treating acute and chronic pain.

Treatment of osteoarthritis: A focus on nimesulide

Widespread osteoarthritis (OA) and high related disability rates determine not only the medical, but also socioeconomic significance of this disease. In recent years there has been a tendency to increase the incidence of OA. According to the current concepts, inflammation plays a leading role in the development of chronic pain in OA, which requires adequate anti-inflammatory therapy. There are data on the diverse mechanisms of action of the nonsteroidal anti-inflammatory drug (NSAID) nimesulide and results from clinical trials and metaanalyses, which confirm its efficacy and good tolerability. Several clinical trials covering 22,938 patients with OA have indicated that the overall frequency of adverse reactions (AR) from nimesulide is 8.2%. There is evidence on the efficacy and good tolerability of topical NSAIDs, as well as combination drugs containing glucosamine sulfate and chondroitin sulfate, the use of which together with oral drugs enhances the efficiency of treatment without increasing the risk of ARs.

Highly sensitive determination of non-steroidal anti-inflammatory drug nimesulide using electrochemically reduced graphene oxide nanoribbons

Preparation of an electrochemically reduced graphene nanoribbon (ER-GONR) film modified screen-printed carbon electrode for the highly sensitive determination of nimesulide.

Crosstalk Among Disrupted Glutamatergic and Cholinergic Homeostasis and Inflammatory Response in Mechanisms Elicited by Proline in Astrocytes.

Hyperprolinemias are inherited disorder of proline (Pro) metabolism. Patients affected may present neurological manifestations, but the mechanisms of neural excitotoxicity elicited by hyperprolinemia are far from being understood. Considering that the astrocytes are important players in neurological disorders, the aim of the present work was to study the effects 1 mM Pro on glutamatergic and inflammatory parameters in cultured astrocytes from cerebral cortex of rats, exploring some molecular mechanisms underlying the disrupted homeostasis of astrocytes exposed to this toxic Pro concentration. We showed that cortical astrocytes of rats exposed to 1 mM Pro presented significantly elevated extracellular glutamate and glutamine levels, suggesting glutamate excitotoxicity. The excess of glutamate elicited by Pro together with increased glutamate uptake and upregulated glutamine synthetase (GS) activity supported misregulated glutamate homeostasis in astrocytic cells. High Pro levels also induced production/release of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6. We also evidenced misregulation of cholinergic anti-inflammatory system with increased acetylcholinesterase (AChE) activity and decreased acetylcholine (ACh) levels, contributing to the inflammatory status in Pro-treated astrocytes. Our findings highlighted a crosstalk among disrupted glutamate homeostasis, cholinergic mechanisms, and inflammatory cytokines, since ionotropic (DL-AP5 and CNQX) and metabotropic (MCPG and MPEP) glutamate antagonists were able to restore the extracellular glutamate and glutamine levels; downregulate TNFα and IL6 production/release, modulate GS and AChE activities; and restore ACh levels. Otherwise, the non-steroidal anti-inflammatory drugs nimesulide, acetylsalicylic acid, ibuprofen, and diclofenac sodium decreased the extracellular glutamate and glutamine levels, downregulated GS and AChE activities, and restored ACh levels in Pro-treated astrocytes. Altogether, our results evidence that the vulnerability of metabolic homeostasis in cortical astrocytes might have important implications in the neurotoxicity of Pro.