Carrageenan-induced Paw Edema Assay Research Articles

Acute toxicity and anti-inflammatory activity of bis-thiourea derivatives

Abstract In the current work, bis-thiourea derivatives have been synthesized through condensation reaction between isothiocynates and diamines in dry acetone to form SK1 (1,2-bis(N-benzoylthioureido) benzene), SK2 (1,3-bis(N-benzoylthioureido) benzene), and SK3 (1,4-bis(N-benzoylthioureido) benzene). The structures of new synthesized derivatives were confirmed through melting point and spectroscopic technique such as 1HNMR only. The synthesized compounds were assessed for acute toxicity test and are proved free of toxicity. The derivatives were further tested as anti-inflammatory agents by in vitro lipoxygenase enzyme inhibition studies, molecular docking, and in vivo carrageenan-induced paw edema assay, and histamine-induced edema test. The overall observations presented that compounds SK1 and SK3 possess promising anti-inflammatory potential, while compound SK2 is found to be a good anti-inflammatory agent.

A Study on Justicia flava (Forssk.) Vahl.: Pharmacognostic Characterization and Antiplasmodial and Anti-Inflammatory Properties of the Leaves

Justiciaflava (Acanthaceae) is utilized in folk medicine for treating malaria, inflammatory conditions, and respiratory infections. In this study, the antiplasmodial and anti-inflammatory activities of the 70% hydroalcoholic leaf extract of J. flava (JFE) were investigated. The antiplasmodial activity was assessed by the suppressive test and Rane’s curative test. The carrageenan-induced paw oedema assay in chicks was used to assess the anti-inflammatory activity. In Rane’s curative test, JFE significantly suppressed the parasitaemia level ( p < 0.0001) by 56.85%, 63.85%, and 77.85% at 30, 100, and 300 mg·kg−1, respectively. The highest suppression of parasitaemia in the suppressive test was 51.31% at 300 mg·kg−1. The extract prolonged the survival time of infected mice significantly and prevented weight loss and hypothermia which are cardinal signs of P. berghei-induced infection. JFE inhibited foot oedema in chicks with a maximum percentage inhibition of 54.00 ± 5.20%. Pharmacognostic characterization of J. flava revealed it to have simple, petiolate, oppositely arranged leaves which are broadly lanceolate in shape with entire to slightly crenate margins. Microscopy of the leaf showed wavy-walled epidermal cells, diallelocytic stomata, abundant clothing and secretory trichomes, pitted vessels, collateral conjoint vascular bundles, cystoliths, and prismatic calcium oxalate crystals. Flavonoids, tannins, phytosterols, triterpenoids, coumarins, alkaloids, and saponins were detected in the plant. This study has provided important parameters for the correct identification of J. flava and given the scientific credence to its antimalarial and anti-inflammatory effects in traditional medicine. J. flava is a potential source of new antimalarial and anti-inflammatory compounds.

Synthesis of Tetracyclic Spirooxindolepyrrolidine-Engrafted Hydantoin Scaffolds: Crystallographic Analysis, Molecular Docking Studies and Evaluation of Their Antimicrobial, Anti-Inflammatory and Analgesic Activities

In a sustained search for novel potential drug candidates with multispectrum therapeutic application, a series of novel spirooxindoles was designed and synthesized via regioselective three-component reaction between isatin derivatives, 2-phenylglycine and diverse arylidene-imidazolidine-2,4-diones (Hydantoins). The suggested stereochemistry was ascertained by an X-ray diffraction study and NMR spectroscopy. The resulting tetracyclic heterocycles were screened for their in vitro and in vivo anti-inflammatory and analgesic activity and for their in vitro antimicrobial potency. In vitro antibacterial screening revealed that several derivatives exhibited remarkable growth inhibition against different targeted microorganisms. All tested compounds showed excellent activity against the Micrococccus luteus strain (93.75 µg/mL ≤ MIC ≤ 375 µg/mL) as compared to the reference drug tetracycline (MIC = 500 µg/mL). Compound 4e bearing a p-chlorophenyl group on the pyrrolidine ring exhibited the greatest antifungal potential toward Candida albicans and Candida krusei (MIC values of 23.43 µg/mL and 46.87 µg/mL, respectively) as compared to Amphotericin B (MIC = 31.25 and 62.50 µg/mL, respectively). The target compounds were also tested in vitro against the lipoxygenase-5 (LOX-5) enzyme. Compounds 4i and 4l showed significant inhibitory activity with IC50 = 1.09 mg/mL and IC50 = 1.01 mg/mL, respectively, more potent than the parent drug, diclofenac sodium (IC50 = 1.19 mg/mL). In addition, in vivo evaluation of anti-inflammatory and analgesic activity of these spirooxindoles were assessed through carrageenan-induced paw edema and acetic acid-induced writhing assays, respectively, revealing promising results. In silico molecular docking and predictive ADMET studies for the more active spirocompounds were also carried out.

Phytochemical characterization, in vivo and in vitro biological activities of Atriplex lindleyi Moq. subsp. inflata

Ethnopharmacological relevanceAtriplex lindleyi Moq. subsp. inflata from Algeria is known for its traditional use for therapeutic properties. Aim of the studyWe analysed quantitatively and qualitatively the infusion and diethyl ether, n-butanol and ethereal extracts of A. lindleyi, as well as assessed their acute toxicity, anti-inflammatory, antinociceptive and antioxidant activities. Materials and methodsThe total phenolic and flavonoid contents were quantified. The anti-inflammatory effect was assessed using a carrageenan-induced paw oedema assay and the antinociceptive effect was evaluated using an acetic acid-induced writhing method. In addition, antioxidant activity was examined by three tests: DPPH, reducing power and phenanthroline assays. ResultsThe quantity of total phenols in the roots of A. lindleyi was larger than in the aerial parts, while the amounts of total flavonoids were larger in the aerial parts than in the roots. The HPLC profiles allowed us to identify 32 compounds belonging to the phenolic acid and flavonoid classes. Intraperitoneal administration of the infusions and phenolic extracts in mice did not cause any symptoms of toxicity or mortality. Results revealed that the aerial parts and roots of A. lindleyi had potential anti-inflammatory and antinociceptive activities and were higher than diclofenac and paracetamol, respectively. The diethyl ether extract (DEE) of aerial parts and roots showed the greatest antioxidant activity in comparison to the other tested extracts. ConclusionThe phenolic composition attempted to be identified by HPLC confers this plant's pharmacological potential. The antioxidant potential may be due to active ingredients, including ascorbic acid, in the two studied parts of A. lindleyi, which is a bioactive molecule with strong antioxidant properties. Also, detecting salicylic acid, the active antinociceptive ingredient of aspirin, gallic and ferulic acids may justify this subspecies' antinociceptive and anti-inflammatory potentials.

HPLC, FTIR and GC-MS Analyses of Thymus vulgaris Phytochemicals Executing In Vitro and In Vivo Biological Activities and Effects on COX-1, COX-2 and Gastric Cancer Genes Computationally.

Medicinal plants have played an essential role in the treatment of various diseases. Thymus vulgaris, a medicinal plant, has been extensively used for biological and pharmaceutical potential. The current study was performed to check the biopotential of active biological compounds. The GC-MS analysis identified 31 compounds in methanolic crude extract, among which thymol, carvacrol, p-cymene, and eugenol are the main phytoconstituents present in T. vulgaris. The HPLC analysis quantified that flavonoids and phenolic acids are present in a good concentration in the active fraction of ethyl acetate and n-butanol. FTIR confirmed the presence of functional groups such as phenols, a carboxylic group, hydroxy group, alcohols, and a benzene ring. Among both fractions, ethyl acetate showed high antioxidant activity in the DPPH (84.1 0.88) and ABTS (87.1 0.89) assays, respectively. The anti-inflammatory activity of the fractions was done in vitro and in vivo by using a carrageenan-induced paw edema assay, while the hexane-based extract showed high anti-inflammatory activity (57.1 0.54) in a dose-response manner. Furthermore, the lead compound responsible for inhibition in the denaturation of proteins is thymol, which exhibits the highest binding affinity with COX1 (-6.4 KJ/mol) and COX2 (-6.3 KJ/mol) inflammatory proteins. The hepatotoxicity analysis showed that plant-based phytoconstituents are safe to use and have no toxicity, with no necrosis, fibrosis, and vacuolar degeneration, even at a high concentration of 800 mg/kg body weight. Furthermore, the in silico analysis of HPLC phytochemical compounds against gastric cancer genes showed that chlorogenic acid exhibited anticancer activity and showed good drug-designing characteristics. Thrombolysis and hemolysis are the major concerns of individuals suffering from gastric cancer. However, the T. vulgaris fractions showed thrombolysis from 17.6 to 5.4%; similarly, hemolysis ranged from 9.73 to 7.1% at a concentration of 12 mg/mL. The phytoconstituents present in T. vulgaris have the potential for multiple pharmacological applications. This should be further investigated to isolate bioactive compounds that can be used for the treatment of different ailments.

Evaluation of Phytochemistry and Pharmacological Properties of Alnus nitida.

In the current study, the anti-inflammatory and analgesic potential of Alnus nitida (leaves and fruits) was evaluated in the Sprague-Dawley rat. Traditionally, A. nitida was used for the treatment of inflammatory ailments. However, A. nitida leaves and fruits have not been yet reported regarding any potential medicinal effects. Leaves/fruits of A. nitida were extracted with methanol and fractionated to attain n-hexane, chloroform, ethyl acetate and aqueous fractions. These extracts were then evaluated for in vivo analgesic and anti-inflammatory potential. For in vivo anti-inflammatory activity, carrageenan-induced paw edema assay, Freunds’ complete adjuvant-induced edema, xylene-induced ear edema and histamine-induced paw edema models were used in rats, which showed significant (p < 0.01) reduction (70–80%) in edema in comparison of inflammatory controls. On other hand, for the analgesic assessment, hot plate assay and acetic acid-induced writhing tests were used, which showed a significant (p < 0.01) rise in latency time (40–60%) as compared with pain-induced controls. These results were comparable with standard drugs in a concentration-dependent manner and no mortality or toxicity was observed during all experiments. Then, for the identification of chemical constituents gas chromatography–mass spectrometry (GC-MS) analysis was performed, which indicated the presence of neophytadiene, 3,7,11,15-Tetramethyl-2-hexadecen-1-ol, phytol and vitamin E, justifying the use of A. nitida to treat inflammatory disorders.

Discovery of benzothiazole-based thiazolidinones as potential anti-inflammatory agents: anti-inflammatory activity, soybean lipoxygenase inhibition effect and molecular docking studies

ABSTRACT Despite the greatest achievement in the development of anti-inflammatory agents in the last two decades, the current clinical drugs suffer from a variety of complications in community settings and hospital. There is still an urgent need to design novel molecules with better safety profile and with different molecular targets from those in current clinical use. The aim of this research was to discover a series of benzothiazole-based thiazolidinones with lipoxygenase (LOX) inhibitory activity as a mechanism of anti-inflammatory action. Carrageenan-induced mouse foot paw oedema assay was carried out to determine the anti-inflammatory activity, while LOX inhibition was examined through the conversion of sodium linoleate to 13-hydroperoxylinoleic acid. Molecular docking studies were performed using AutoDock 4.2 software. The anti-inflammatory activity of the title compounds was determined in a range of 18.4%-69.57%, where compound #3 was found to be the most potent (69.57%) and also to be more active than the reference drug indomethacin (47%). Moreover, compound #3 showed the highest LOX inhibitory activity with IC50 of 13 μM being less potent to that of the reference NDGA (IC50 = 1.3 μM). Compound #3 has been identified as lead compound for further modification in an attempt to improve anti-inflammatory and LOX inhibitory activities.

Cedrus atlantica (Endl.) Manetti ex Carrière Essential Oil Alleviates Pain and Inflammation with No Toxicity in Rodent

Cedrus atlantica (Endl.) Manetti ex Carrière is an endemic tree with spiritual value, and it was used since immemorial time in folk medicine. The present study aims to evaluate the anti-inflammatory (carrageenan-induced paw edema and formalin tests) and analgesic effects (hot plate and acetic acid writhing tests) of the cedarwood essential oil, as well as inspect any toxicity (acute toxicity), using several in vivo assays. Following the acetic acid writhing test and the hot plate test, the EO presented an excellent analgesic effect compared to the controls, especially with the dose of 50 mg/kg. Similar results were found while assessing the anti-inflammatory potential in the carrageenan-induced paw edema and formalin assays. The acute toxicity assessment and the subsequent monitoring of the animals, the biochemical analysis, and the relative organ weight, demonstrated a total safety of the EO. The GC/MS analysis of the composition revealed that the major compounds contained in this EO are beta-himachalene (51.95%), followed by alpha-himachalene (15.82%), and gamma-himachalene (12.15%). This study supports the usage of this tree EO to alleviate pain and inflammation.

Anti-Inflammatory Potential of the Oleoresin from the Amazonian Tree Copaifera reticulata with an Unusual Chemical Composition in Rats.

Copaifera reticulata Ducke is a popularly known species known as copaíba that is widely spread throughout the Amazon region. The tree yields an oleoresin which is extensively used in local traditional medicine mainly as an anti-inflammatory and antinociceptive agent. The aim of the present study was to assess the anti-inflammatory potential of this oleoresin obtained from a national forest in the central Amazon which presented an unusual chemical composition. The chemical composition of volatile compounds of oleoresin was analyzed by gas chromatography-mass spectrometry. The acute toxicity assay was performed with a single dose of 2000 mg/kg. The anti-inflammatory potential was evaluated by carrageenan-induced paw edema and air pouch assays using four different C. reticulata oleoresin concentrations (10, 100, and 400 mg/kg). The exudate was evaluated for nitrite concentration through the colorimetric method and for TNF-α, IL-1β, and PGE2 by ELISA. C. reticulata oleoresin collected in the Amazonian summer contained six major sesquiterpene compounds (β-bisabolene, cis-eudesma-6,11-diene, trans-α-bergamotene, β-selinene, α-selinene, and β-elemene) and was nontoxic at a dose of 2000 mg/kg, showing low acute toxicity. Different from oleoresin obtained from other sites of the Brazilian Amazon, the major volatile compound found was β-Bisabolene with 25.15%. This β-Bisabolene-rich oleoresin reduced the formation of paw edema induced by carrageenan and reduced the global number of cells in the air pouch assay, as well as exudate volume and nitrite, TNF-α, IL-1β, and prostaglandin E2 levels (p < 0.05). C. reticulata oleoresin with a high β-Bisabolene concentration showed anti-inflammatory activity, reducing vascular permeability and consequently edema formation, and thus reducing cell migration and the production of inflammatory cytokine, confirming its traditional use by local Amazonian communities.

New quinoline/1,2,4-triazole hybrids as dual inhibitors of COX-2/5-LOX and inflammatory cytokines: Design, synthesis, and docking study

• Novel quinoline/1,2,4-triazole hybrids 6a-i and 7a-j as anti-inflammatory agents has been designed and synthesized. • Compounds 6e, 6i , and 7e showed potent COX-2 inhibition. • Compounds 6e and 6i showed potent anti-inflammatory activity relative to celecoxib with high gastric safety profile. • 6e, 6i , and 7e showed a significant decrease in the pro-inflammatory cytokines. • Docking study revealed high binding affinities toward COX-2 and 5-LOX with the formation of essential hydrogen bonds. A novel series of 19 quinoline/1,2,4-triazole hybrid 6a-i and 7a-j was synthesized and evaluated in vitro as dual COX-2/5-LOX inhibitors. Compounds 6e, 6i , and 7e displayed the highest potency and selectivity for inhibiting COX-2 activity (IC 50 = 7.25, 8.13, and 8.48 nM, respectively; selectivity index (COX-1/COX-2) = 44.89, 30.30, and 33.47, respectively) in comparison to celecoxib (COX-2 IC 50 = 42.60 nM; selectivity index (SI) = 8.05). The anti-inflammatory activity of the newly synthesized compounds was further examined in vivo using a carrageenan induced paw edema assay. Interestingly, the in vitro findings of the COX inhibitory assay were consistent with the in vivo assay. Moreover, 6e, 6i , and 7e showed a substantial reduction in serum concentrations of PGE 2 , TNF-α, IL-6. Molecular docking analysis of compounds 6e , 6 f, 6i, 7e , and 7f revealed high binding affinities toward COX-2 compared to COX-1, which was matched with the experimental results. In addition, these compounds exhibited different binding orientations into the active site of COX-2, which were dependent on the type of substitutions on N4 of the triazole ring. Among the tested derivatives, compounds 6e, 6i and 7e which showed high selectivity to COX-2, exhibited hydrogen bonding interactions with key amino acids in COX-2 such as Arg120, Arg513, and/or Glu524. In addition, the tested compounds also showed multiple hydrogen bonds with the Arg101, Val110, Arg138 or His130 in 5-LOX. These findings show, taken together, that those derivatives are good leads to potential anti-inflammatory agents with lowest gastric damage.

The Hydroethanolic Stem Bark Extract of Tieghemella heckelii (A.Chev.) Pierre ex Dubard (Sapotaceae) Produced N-Methyl-D-Aspartate (NMDA) Receptor-Dependent Analgesia and Attenuates Acute Inflammatory Pain via Disruption of Oxidative Stress.

BackgroundTieghemella heckelii stem bark is used in African traditional medicine to treat inflammatory pain conditions. However, these biological actions of the plant have not been proven. This study investigates the phytochemical composition and the mechanisms of analgesic and anti-inflammatory actions of the hydroethanolic stem bark extract of T. heckelii (THBE). Methods Phytochemical composition of THBE was investigated using qualitative and quantitative phytochemical analyses. Anti-inflammatory activity was evaluated using the carrageenan-induced paw oedema assay. Analgesic activity was evaluated using hot plate and acetic acid-induced writhing assays. Mechanism of analgesic action was determined using pharmacological antagonist such as naloxone, atropine, flumazenil, nifedipine, or ketamine. Test agents were administered orally as follows: Tween 80 (5%) (control), diclofenac sodium (DS) 10/tramadol 9 mg/kg (standard), or THBE 10, 100, and 450 mg/kg. Glutathione peroxidase (GPx), superoxide dismutase (SOD), and lipid peroxidation levels were also measured. Results THBE which contained 58.45% saponins, 229.04 ± 0.049 GAE mg/g phenolic compounds,and 0.482 ± 0.0028 QE mg/g flavonoids produced (p < 0.5) anti-inflammatory effect of 56.22% and analgesia of 330 ± 72% and 50.4% in the hot plate and writhing assays, respectively, at 10 mg/kg and inhibited oxidative stress by GPx and SOD elevation in rats during inflammation. Ketamine significantly blocked the analgesia of THBE, indicating NMDA receptor-dependent analgesic action. Whereas, naloxone, atropine, nifedipine, and flumazenil could not antagonize the analgesic action of THBE. Conclusion These results show that THBE produced potent anti-inflammatory effect via disruption of oxidative stress and also generated NMDA receptor-dependent analgesia.

Phytochemical profiling, in vitro and in vivo anti-inflammatory, analgesic and antipyretic potential of Sesuvium sesuvioides (Fenzl) Verdc. (Aizoaceae).

Sesuvium sesuvioides (Fenzl) Verdc is traditionally used in the treatment of inflammatory diseases such as arthritis and gout The aim of present study was to assess the possible anti-inflammatory, analgesic and antipyretic potential of the methanol extract of Sesuvium sesuvioides (SsCr) to prove scientifically its folklore use in the inflammatory diseases and to screen its total antioxidant capacity by multiple methods and phytocompounds by GC-MS. The preliminary phytochemical studies showed the presence of phenols, flavonoids, glycosides, coumarin, terpenoids, saponins, fats and carbohydrates in crude extract. The total phenolic contents (27.31 ± 0.28mg GAE/g) and total flavonoids (3.58 ± 0.12 mgRE/g) values were observed. The antioxidant capacity of SsCr showed significant DPPH, ABTS, CUPRAC, FRAP, PBD and metal chelating results. GC-MS analysis displayed the phytoconstituents with anti-inflammatory potentials such as 2-methoxy-4-vinylphenol, vanillin, umbelliferone, methyl ferulate, palmitoleic acid, methyl palmitate and phytol. SsCr presented noteworthy HRBC membrane stability with maximum inhibition of cell hemolysis (47.79%). In carrageenan-induced hind paw edema assay result showed dose-dependent anti-inflammatory action. SsCr presented significant (p < 0.05) analgesic activity in hot-plate and tail flicking tests similarly it also showed the noteworthy inhibition in pain latency against formalin induced analgesia at 1st and 2nd phases. SsCr reduced the acetic acid-induced writhes at different doses (250, 500 and 750mg). Results of antipyretic activity of SsCr extract were significant at 500 and 750mg. The results of in vitro and in vivo experimental studies verified the anti-inflammatory, analgesic and antipyretic potential of Sesuvium sesuvioides and supported the folklore uses of this plant.

Govaniadine Ameliorates Oxidative Stress, Inflammation, and Kupffer Cell Activation in Carbon Tetrachloride-Induced Hepatotoxicity in Rats.

Liver diseases such as hepatic carcinoma are one of the main health problems worldwide. Herbal drugs are largely used to treat liver injury in the indigenous system of medicine and may provide lead compounds for hepatoprotective drug discovery. The present study is investigated to test the Corydalis govaniana Wall. extract, fraction, and isolate therapeutically active constituents to explore their hepatoprotective, anti-inflammatory, and antioxidant activities. For this purpose, the antioxidant activity of govaniadine, caseadine, caseamine, and protopine was performed by assessing the scavenging events of the stable 2,2-diphenyl-1-picrylhydrazyl. Hepatoprotection of govaniadine was assessed in terms of reduction in serum enzymes (alanine aminotransferase, aspartate transaminase, and alkaline phosphatase) caused by CCl4-induced liver injury in rats and by histopathological techniques. All the compounds showed significant antioxidant activity with a percentage inhibition of 92.2, 86.7, 85.3, and 79.7, respectively, compared to propyl gallate 90.3%. Treatment with govaniadine reduced the serum enzyme level down to normal levels in the CCl4-treated group while inhibiting the increase of malondialdehyde, and the induction of superoxide dismutase and the glutathione level was upregulated. Histopathology showed ∼47% damage to the liver cells in the CCl4-treated group; reduction in this damaged area was found to be better upon using govaniadine. Immunohistochemistry results showed that govaniadine as compared to silymarin has exceedingly decreased the inflammation by halting the CCl4-induced activation of hepatic macrophages. In carrageenan-induced paw edema assay, govaniadine significantly alleviated the edema after 1–5 h at a dose of 20 mg/kg (26.00 and 28.5%), 50 mg/kg (22.05 and 27.0%), and 100 mg/kg (20.02 and 25.30%), respectively. The results of our experiments suggest that govaniadine showed antioxidant and hepatoprotective activity in liver injury. The hepatoprotective function of govaniadine may be associated to the scavenging of the free radical and attenuation of oxidative stress as well as inflammatory responses in the liver. Hence, govaniadine may be a lead compound for the hepatoprotective drug discovery process and further research is needed to find out their molecular mechanism of protection.

Anti-inflammatory and analgesic potential of Amorphophallus commutatus var. wayanadensis and its inhibitory effect on inflammatory mediators in lipopolysaccharide-stimulated macrophages

Background: An edible tuber named Amorphophallus commutatus var. wayanadensis (ACW) is used by the local ethnic communities of Wayanad, India, for hemorrhoids while the health benefits remain unexplored to the scientific community. Objectives: Hence, our study was performed to screen the anti-inflammatory and analgesic activities of ACW. Materials and Methods: Screening for the in vitro anti-inflammatory potential on isolated peritoneal macrophages was performed using nitrite assay, nitroblue tetrazolium assay, tumor necrosis factor (TNF)-α production, and cyclooxygenase (COX) enzyme activity. Carrageenan- and formalin-induced paw edema assays were performed to determine anti-inflammatory potential, while tail immersion assays and acetic acid-induced writhing assays were carried out to explore antinociceptive activity in animal models. Results: Bio-molecular mechanistic investigation to evaluate the in-vitro anti-inflammatory activity confirmed the suppressive effect of MEAC on TNF-α, nitric oxide and COX-2 on lipopolysaccharide stimulated peritoneal macrophages, which benchmarked ACW as a potent medicinal plant against inflammation. Further, the efficacy of methanolic extract of A. commutatus var. wayanadensis (MEAC) as an anti-inflammatory agent in murine anti-inflammatory models was demonstrated by formalin- and carrageenan-induced paw edema assays. Administration of MEAC significantly increased the tail flicking latency in mice and also showed prominent reduction in the number of writhes induced by acetic acid, which establishes the real time application of ACW as an analgesic agent. Conclusion: The vital information regarding in vitro and in vivo action of MEAC provided scientific evidence for its traditional usage for hemorrhoids. Thus, this herbal medicine of ethnic tribes can be translated toward modern medicine for health maintenance and disease prevention.

Anti-Inflammatory and Analgesic Activities of 7-Chloro-4-(Piperazin-1-yl) Quinoline Derivative Mediated by Suppression of InflammatoryMediators Expression in Both RAW 264.7 and Mouse Models

Background: 4-Aminoquinoline derivatives possess various potential biological properties.The introduction of additional piperazine heterocyclic pharmacophoric moiety tends to haveprofound impact in increasing the activity. The present work was undertaken to investigate thein-vitro and in-vivo anti-inflammatory activity as well as the peripheral and central analgesicactivities of compound 1-(4-(7-chloroquinoline-4-yl)piperazin-1-yl)-2-(4-phenylpiperazin-1-yl)ethanone (5) in experimental models. Methods: The percentage inhibition of the lipopolysaccharide induced NO release of 7-chloro-4-(piperazin-1-yl)quinoline derivatives 1-9 was determined in RAW 264.7 murine macrophagemodel. Western blot analysis was performed to evaluate the effect of compound 5 on proteinexpression of inducible nitric oxide synthase (iNOS). Gene expression of inflammatory markerswas evaluated using real-time polymerase chain reaction. The peripheral and central analgesicactivities of compound 5 were evaluated in mice using writhing and hot-plate tests, respectively.Anti-inflammatory activity was assessed using carrageenan-induced paw edema assay in miceand serum NO and COX-2 levels were measured. Results: Compound 5 demonstrated the highest NO inhibitory activity that was accompaniedby inhibition of iNOS protein expression and decreased gene expression levels of inflammatorymarkers. It revealed a potential peripheral analgesic effect through inhibition of abdominalwrithing in mice treated with doses of 15 and 30 mg/kg and its effect was comparable to diclofenacsodium. Compound 5 possessed an analgesic activity starting from 15 min post administrationand reached its peak at 45 min which was significantly higher than that of tramadol hydrochloridesuggesting its potential as central analgesic agent. It also showed percentage of inhibition ofedema of 34, 50 and 64% at 1, 2, and 3 h respectively, post carrageenan challenge together with asignificant decrease in serum NO and COX-2 levels. Conclusion: The remarkable anti-inflammatory and analgesic activities of compound 5 couldbe attributed to the advantageous introduction of the heterocyclic 7-chloro-4-(piperazin1-yl)quinoline scaffold incorporated with N-phenylpiperzine functional groups linked together withthe ethanone pharmacophoric chain.

Novel approach of multi-targeted thiazoles and thiazolidenes toward anti-inflammatory and anticancer therapy—dual inhibition of COX-2 and 5-LOX enzymes

It is well established that cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) play a vital role in the initiation and progression of inflammatory reactions. Hence, thiazole and thiazolidene-based pharmacophore molecules were synthesized to obtain dual COX-2 and 5-LOX inhibitory activity. The synthesis of target compounds has been achieved by a novel green strategy. In vitro COX-1, COX-2, and 5-LOX evaluation of these molecules have shown the potential for an improved anti-inflammatory profile. Most promising compound among the series (2-(diphenylamino)-4-(4-nitrophenyl)thiazol-5-yl)(naphthalen-1-yl)methanone 7h (IC50 = 0.07 ± 0.02 μM) showed equivalent COX-2 inhibitory potency as that of positive control etoricoxib (IC50 = 0.07 ± 0.01 μM) and an enhanced selectivity index of 115.14. Compound 7h exhibited 5-LOX IC50 of 0.29 ± 0.09 μM and reference drug zileuton showed IC50 of 0.15 ± 0.05 μM. In vivo studies of 7h including carrageenan-induced paw edema assay (63% inhibition of paw edema), antiulcer studies, biochemical assays, qRT-PCR analysis, and anticancer studies indicated that the present study has identified a good lead compound for the development of a potential anti-inflammatory drug having improved gastric safety profile.

Ailanthus altissima (Miller) Swingle seed oil: chromatographic characterization by GC-FID and HS-SPME-GC-MS, physicochemical parameters, and pharmacological bioactivities.

This study aimed to identify the physicochemical and the chemical properties of Ailanthus altissima (Miller) Swingle seed oil and to evaluate its in vitro antioxidant and antibacterial activities and in vivo analgesic and anti-inflammatory activities. The fatty acids' composition was determined using GC-FID. The oil was screened for antioxidant activity by DPPH test. The analgesic and anti-inflammatory activities were determined using the acetic acid writhing test in mice and the carrageenan-induced paw edema assay in rats, respectively. Volatile compounds were characterized by HS-SPME-GC-MS. A. altissima produces seeds which yielded 17.32% of oil. The seed oil was characterized by a saponification number of 192.6mg KOH∙g of oil, a peroxide value of 11.4meq O2∙kg of oil, a K232 of 4.04, a K270 of 1.24, and a phosphorus content of 126.2ppm. The main fatty acids identified were palmitic (3.06%), stearic (1.56%), oleic (38.35%), and linoleic acids ones (55.76%). The main aroma compounds sampled in the headspace were carbonyl derivatives. The oil presents an important antioxidant activity (IC50 = 24.57μg/mL) and a modest antimicrobial activity. The seed oil at 1g/kg showed high analgesic (91.31%) and anti-inflammatory effects (85.17%). The presence of high levels of unsaturated fatty acids and the noteworthy antioxidant capacity of the seed oil can hypothesize its use as an analgesic and anti-inflammatory agent.

Analgesic, Anti-inflammatory and Antihyperlipidemic Activities ofKnema attenuata, an Endemic Plant of Southern India

The analgesic, anti-inflammatory and antihyperlipidemic activities ofKnema attenuata(Myristicaceae), one of the ingredients of ‘Ashwagandhadhi nei (medicated ghee)’ used in the treatment of spleen disorders according to the Ayurvedic herbal medicine tradition are investigated using suitable in vivo models such as hot plate method, tail immersion test, carrageenan-induced paw oedema assay and Triton X-100-induced hyperlipidemic assay. The seed and aril (a fleshy to hard structure which develops from the funicle or ovule after fertilisation and encloses all or part of the seed) extracts were used. Both the extracts showed significant dose dependent analgesic activity up to 400 mg/kg. The dose dependent ethanolic extract of aril exhibited higher latency to hot plate and tail withdrawal reflex proving its analgesic property. Anti-inflammatory activity was demonstrated after 3 h of oedema induction. The paw volume (ml) decreased from 1.36 ± 0.05 in control to 0.73 ± 0.02 and 0.78 ± 0.02 respectively in groups treated with ethanol extract of aril and seed. The extracts of seed and aril administered at a dose of 400 mg/kg lowered the serum TC (total cholesterol), TG (triglycerides), LDL (low density lipoprotein) and VLDL (very low density lipoprotein) compared to hyperlipidemic groups. A significant decrease in the atherogenic index (a strong marker to predict the risk of atherosclerosis and coronary heart disease) was observed in animals supplemented withKnema attenuatawhich suggests an athero protective/ cardio protective potential of this plant extract. We conclude that ethanolic extract ofKnema attenuatahas significantly good analgesic, anti-inflammatory and antihyperlipidemic activity. It could be a new source of natural analgesic agent and an alternative natural source of antiinflammatory and cardio protective agents.

Synthesis, in vitro and in vivo biological evaluation, COX-1/2 inhibition and molecular docking study of indole-N-acylhydrazone derivatives

The objective of this work was to obtain and evaluate anti-inflammatory in vitro, in vivo and in silico potential of novel indole-N-acylhydrazone derivatives. In total, 10 new compounds (3a–j) were synthesized in satisfactory yields, through a condensation reaction in a single synthesis step. In the lymphoproliferation assay, using mice splenocytes, 3a and 3b showed inhibition of lymphocyte proliferation of 62.7% (±3.5) and 50.7% (±2), respectively, while dexamethasone presented an inhibition of 74.6% (±2.4). Moreover, compound 3b induced higher Th2 cytokines production in mice splenocytes cultures. The results for COX inhibition assays showed that compound 3b is a selective COX-2 inhibitor, but with less potency when compared to celecoxib, and compound 3a not presented selectivity towards COX-2. The molecular docking results suggest compounds 3a and 3b interact with the active site of COX-2 in similar conformations, but not with the active site of COX-1, and this may be the main reason to the COX-2 selectivity of compound 3b. In vivo carrageenan-induced paw edema assays were adopted for the confirmation of the anti-inflammatory activity. Compound 3b showed better results in suppressing edema at all tested concentrations and was able to induce an edema inhibition of 100% after 5 h of carrageenan injection at the 30 mg kg−1 dosage, corroborating with the COX inhibition and lymphoproliferation results. I addition to our experimental results, in silico analysis suggest that compounds 3a and 3b present a well-balanced profile between pharmacodynamics and pharmacokinetics. Thus, our preliminary results revealed the potentiality of a new COX-2 selective derivative in the modulation of the inflammatory process.

Attenuation of inflammatory responses by (+)-syringaresinol via MAP-Kinase-mediated suppression of NF-\u03baB signaling in vitro and in vivo

We examined the anti-inflammatory effects of (+)-syringaresinol (SGRS), a lignan isolated from Rubia philippinensis, in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells using enzyme-based immuno assay, Western blotting, and RT-PCR analyses. Additionally, in vivo effects of SGRS in the acute inflammatory state were examined by using the carrageenan-induced hind paw edema assay in experimental mice. As a result, treatment with SGRS (25, 50, and 100 μM) inhibited protein expression of lipopolysaccharide-stimulated inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear factor kappa B (NF-κB) as well as production of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), and interleukin-6 (IL-6) induced by LPS. Moreover, SGRS also reduced LPS-induced mRNA expression levels of iNOS and COX-2, including NO, PGE2, TNF-α, IL-1β, and IL-6 cytokines in a dose-dependent fashion. Furthermore, carrageenan-induced paw edema assay validated the in vivo anti-edema effect of SGRS. Interestingly, SGRS (30 mg/kg) suppressed carrageenan-induced elevation of iNOS, COX-2, TNF-α, IL-1β, and IL-6 mRNA levels as well as COX-2 and NF-κB protein levels, suggesting SGRS may possess anti-inflammatory activities.