In-vivo Anti-inflammatory Studies Research Articles

Design, synthesis, biological evaluation, and in silico studies of quinoxaline derivatives as potent p38α MAPK inhibitors.

A new series of quinoxaline derivatives possessing the hydrazone moiety were designed, synthesized, and screened for in-vitro anti-inflammatory activity by the bovine serum albumin (BSA) denaturation technique, and for antioxidant activity, by the (2,2'-diphenyl-1-picrylhydrazyl(DPPH) radical scavenging assay. The synthesized compounds were also tested for p38α mitogen-activated protein (MAP) kinase inhibition. The in-vivo anti-inflammatory activity was assessed by the carrageenan-induced rat paw edema inhibition method. All the compounds (4a-n) exhibited moderate to high in-vitro anti-inflammatory activity. Compound 4a displayed the highest inhibitory activity in the BSA assay (83.42%) in comparison to the standard drug diclofenac sodium (82.90%), while 4d exhibited comparable activity (81.87%). The DPPH assay revealed that compounds 4a and 4d have free radical scavenging potential (74.70% and 74.34%, respectively) comparable to the standard butylated hydroxyanisole (74.09%). Furthermore, the p38α MAP kinase inhibition assay demonstrated that compound 4a is highly selective against p38α MAP kinase (IC50 = 0.042) in comparison to the standard SB203580 (IC50 = 0.044). The five most active compounds (4a-4d and 4f) with good in-vitro profiles were selected for in-vivo anti-inflammatory studies. Compounds 4a and 4d were found to display the highest activity (83.61% and 82.92% inhibition, respectively) in comparison to the standard drug diclofenac sodium (82.65% inhibition). These compounds (4a and 4d) also exhibited better ulcerogenic and lipid peroxidation profiles than diclofenac sodium. The molecular docking and molecular dynamics simulation studies were also performed and found to be in agreement with the p38α MAP kinase inhibitory activity.

In-Vivo and In-Vitro Anti-Inflammatory Activities of the Aqueous Extract of Di-Herbal Formulation (Euphorbia hirta and lactuca virosa)

Euphorbia hirta and Lactuca virosa are medicinal plants that have been used in the cure and treatment of various diseases and for health care. This study aims at evaluating the anti-inflammatory activities of the aqueous extracts of Euphorbia hirta and Lactuca virosa plants. Albumin and histamine induced inflammation in mice and xylene induced ear edema were used for the in-vivo anti-inflammatory studies. Erythrocyte membrane stabilization and inhibition of protein denaturation assays were used for the in-vitro anti-inflammatory studies. Combined doses of 100 mg/kg Euphorbia hirta and 50 mg/kg Lactuca virosa, 100 mg/kg Euphorbia hirta and 100 mg/kg Lactuca virosa, 100 mg/kg Lactuca virosa only and 10 mg/kg diclofenac significantly reduced inflamed paw in mice (P<0.05) compared to control in albumin and histamine induced inflammatory test. Combined doses of 50 mg/kg Euphorbia hirta and 100 mg/kg Lactuca virosa,100 mg/kg Euphorbia hirta, 100mg/kg Lactuca virosa significantly reduced xylene induced inflammation (P<0.001) compared to control. The extracts at 1 mg/ml, 2 mg/ml and 3 mg/ml significantly inhibited protein denaturation (P<0.001) and heat induced hemolysis of erythrocytes (P<0.0001). The plant extract of Euphorbia hirta and lactuca virosa possesses in-vivo and in-vivo anti-inflammatory effects.
 Keywords: Euphorbia hirta, Lactuca virosa, anti-inflammatory, protein denaturation

Liposomal drug delivery of Corchorus olitorius leaf extract containing phytol using design of experiment (DoE): In-vitro anticancer and in-vivo anti-inflammatory studies

Phytol, a pharmacologically active compound present in Corchorus olitorius leaf exhibit a range of activity including anti-inflammatory, antioxidant, anticancer, hepatoprotective etc. However, phytol is poorly soluble and absorbed through the intestine wall, therefore the aim of this study is to develop liposomal drug delivery of Corchorus olitorius leaf extract with an average particle size below 150 nm and drug loading efficiency of ≥ 85 %. The impact of different process parameters and material attributes were studied on the average particle size and polydispersity of liposomal batches using design of experiment (DoE). Corchorus olitorius leaf extraction was performed using maceration method and characterised using GC–MS. Liposomal batches of Corchorus olitorius leaf extract were characterized using Malvern zetasizer, transmission electron microscopy (TEM) and UV spectroscopy. The in-vivo anti-inflammatory study of the liposomal preparation of phytol was evaluated using a rat model and in-vitro cancer cell line study was performed on AML and Leukamia cell lines. GC–MS study data showed that phytol is present in C. olitorius leaf extract. Process parameters and material attributes perspective processing temperature, buffer pH and drug: lipid ratio is found as major parameters affecting the average particle size and PDI value of liposomes. Liposomes were prepared in the range of 80–250 nm and optimized batches of liposomes showed drug entrapment efficiency of 60–88 %. In-vivo anti-inflammatory study showed significant activity for C. olitorius leaf extract against carrageenan induced paw edema, which is significantly increased while delivered through liposomes. In-vitro cancer cell line study data suggests that liposomal delivery of phytol was more active at lower concentration compared to pure phytol, for specific cell lines.

Formulation and Evaluation of Transdermal Gel of Ibuprofen: Use of Penetration Enhancer and Microneedle

The objective of the current study was to develop Ibuprofen (IBP) gel using different polymers individually and in combination and then to select best gel formulation based on various in-vitro evaluation parameters such as bioadhesive strength, gel strength, spreadability, viscosity and drug release study. The selected gel formulation was found to be composed of 1% (w/w) each of hydroxypropyl methylcellulose (HPMC K100M) and sodium carboxy methylcellulose (NaCMC). Two techniques such as chemical method using 1,8-cineole as chemical penetration enhancer (CPE) and physical technique using microneedle were employed to improve IBP permeation across the abdominal skin of rat. Out of the two techniques, the later technique showed higher (2.865-fold) permeation enhancement compared to control. Furthermore, a synergistic effect was also observed when both the techniques were used simultaneously with 3.307-fold increase in permeation enhancement. In-vivo anti-inflammatory study on rats induced with carrageenan paw oedema and analgesic activity investigation by tail flick method in rat model exhibited sustained effect up to 8 h compared to orally treated group. The stability study at room and accelerated conditions for three months did not show any sign of instability. Thus, the developed IBP gel is stable and have potential to illicit both anti-inflammatory and analgesic effect when administered transdermally.

PERFORMANCE OF CURCUMIN IN NANOSIZED CARRIERS NIOSOMES AND ETHOSOMES AS POTENTIAL ANTI-INFLAMMATORY DELIVERY SYSTEM FOR TOPICAL APPLICATION

Curcumin (CUR) is one of the most commonly used herbal product; it shows effective anti-inflammatory and anti-oxidant effects. However, poor aqueous solubility and low permeability are the major challenges in therapeutic application of curcumin. One class of vesicular nanocarriers called “Niosome and ethosome” which have proved to possess distinct advantages were used to encapsulate curcumin and evaluated for their morphology, particle size, zeta potential, entrapment efficiency (EE%) and drug release. They were incorporated into hydroxy propyl methyl cellulose (HPMC15000) gel then, evaluated on the rat skin via inhibition of carrageenan induced rat paw edema. The results showed that the particle size of curcumin loaded niosomes and ethosomes were ranged (317.5±1.91 to 558.3±8.587 nm) and (182.1±5.3 to 354.5±30.03 nm), respectively. Skin permeation studies demonstrated that CUR permeability coefficient through rat kin for gel formulations of loaded vesicles was ~ four times higher as compared to free CUR. The in-vivo anti-inflammatory studies proved that gel formulations of CUR vesicles possessed higher significant inhibition of carrageenan induced rat paw edema when compared to pure curcumin. Accordingly, the results revealed that, curcumin loaded nanovesicels held great potential approaches as anti-inflammatory in topical application.

Synthesis, COX-2 inhibition and metabolic stability studies of 6-(4-fluorophenyl)-pyrimidine-5-carbonitrile derivatives as anticancer and anti-inflammatory agents

In continuation of previous work on cyanopyrimidine derivatives for cancer and inflammation, eighteen novel amino containing cyanopyrimidine derivatives (4a-p) bearing secondary or tertiary amines and alkyl-thio substitution were synthesized by Biginelli condensation reaction followed by nucleophilic substitution reaction. Two compounds 4 g (NSC: 795,754) and 4o (NSC: 798,988) showed excellent anticancer potency against most of the cell lines. Compound 4 g was selected for five dose-assays and the result of tested compound was given by three response parameters i.e. GI50, TGI and LC50 for each cell line. Compound 4 g exhibited superior anticancer activity against ovarian cancer with GI50-value of 0.33 μM and selectivity index of 4.84 in comparison to 5-fluoro uracil (5-FU) which exhibited GI50-value 4.43 μM. During in-vivo anti-inflammatory study, compound 4 g exhibited 87 % inhibition of inflammation at 3 h which was found superior to the standard drugs ibuprofen (78 %) and celecoxib (52 %). The compounds 4 g and 4o exhibiting both anticancer and anti-inflammatory potential were further studied for their biological target studies and in-vitro metabolic stability assessment. The result showed that compounds 4 g and 4o displaying broad anticancer activity were more selective towards COX-2 as compared to COX-1.

SYNTHESIS, COMPUTER AIDED SCREENING AND PHARMACOLOGICAL EVALUATION OF 2/3-SUBSTITUTED-6(4-METHYLPHENYL)-4,5-DIHYDROPYRIDAZIN3(2H)-ONES, AND PYRIDAZINE SUBSTITUTED TRIAZINE

The present research work involved synthesis of some novel pyridazine derivatives and evaluation of their analgesic and anti-inflammatory activities in experimental animals to obtain safer non-steroidal anti-inflammatory drugs (NSAIDs). Friedal craft acylation reaction of succinic anhydride with toluene in the presence of anhydrous aluminum chloride gave 4-(4-methylphenyl)-4-oxo-butanoic acid ( 1) . The aryl propionic acid 1 on reaction with phenyl hydrazine and hydrazine hydrate yielded the pyridazinone derivative 2 and 3 , respectively. Reaction of the compound 3 with phosphorus oxychloride (POCl 3 ) produced the corresponding chloropyridazine derivative 4 . A 4-hydroxymethyl derivative of dihydropyridazinone ( 5) was synthesized by condensing 3 with methanol and formaldehyde (HCHO). The compound 5 on further treatment with guanidine hydrochloride in ethanol gave the pyridazino-triazine ( 6) . The synthesized compounds were investigated for their analgesic activity in mice and anti-inflammatory activity in Wistar albino rats. The molecular, pharmacokinetic and toxicity properties of the synthesized compounds were calculated by Molinspiration and Osiris property explorer software. The results of in-vivo anti-inflammatory studies revealed that the compound. 4 showed maximum inhibition in paw edema volume followed by compound no. 3 while the compound no. 4 exhibited excellent peripheral analgesic activity ( 74%) followed by the compound no. 5 . Compound no. 4 and 5 also showed good central analgesic effect increased the reaction time to 90 minutes. All the title compounds except compound 5 are predicted to be safe by Osiris online software and are likely to have good oral bioavailability as they obey Lipinski’s rule of five for drug likeness.

Synthesis, characterization and anti-inflammatory activity some 1-(4-substituted)-2-(−4-(piperazine-1-yl) bis-thiazole-5-yl) 2-methyl-4-nitro-1h-imidazole-1-yl) ethanone

Novel set of 1-(4-substituted)-2-(−4-(piperazine-1-yl) bis-thiazole-5-yl) 2-Methyl-4-Nitro-1H-imidazole-1-yl) ethanon compounds were synthesized. The adduct of piperazine were synthesized by nucleophilic addition of benzoyl isothiocyanate/substituted benzoyl isothiocyanate in bi-molar quantity at reflux temperature. The target compounds (AR-15a to AR-55a) were synthesized as 1-chloro-3-(2-methyl-4-nitro1H-imidazole-1-yl) propane-2-on (a) was added in bi-imolar quantity to a solution of adduct (substituted-bis-piperazinyl-4-thioyl) benzamide/chlorobenzamide in acetonitrile. The synthesized compounds were characterized by physicochemical parameters and IR, 1H NMR, Mass and elemental spectral analysis. The synthesized compounds were evaluated for their in-vitro anti-inflammatory activity using HRBC membrane stabilization method and in-vivo anti-inflammatory activity by carrageenin-induced rat paw oedema model at 20 mg/kg body weight using diclofenac and ibuprofen as standard drugs. Compounds AR-45a and AR-55a showed maximum in-vitro anti-inflammatory activity membrane stabilization at 65% and 68% protection at 200 μ/mL. However moderate to poor protection was observed by in-vivo anti-inflammatory study in carragenane induced rat hind paw edema.

In-Vitro Assessment and Pharmacodynamics of Nimesulide Incorporated Aloe vera Transemulgel

The aim of the investigation was to prepare nimesulide emulsion for incorporation in Aloe vera gel base to formulate 'nimesulide - Aloe vera transemulgel' (NAE) and to carryout in-vitro assessment and in-vivo anti-inflammatory studies of the product. Although the use of nimesulide is banned for oral administration, due to its potential for inducing hepatotoxicity and thrombocytopenia, the use of nimesulide for topical delivery is prominent in the treatment of many inflammatory conditions including rheumatoid arthritis. The drug loading capacity of transdermal gels is low for hydrophobic drugs such as nimesulide. Nimesulide can be effectively incorporated into emulgels (a combination of emulsion and gel). Aloe vera has a mild anti-inflammatory effect and in the present study Aloe vera gel was formulated and used as a gel base to prepare NAE. The emulgels thus prepared were evaluated for viscosity, pH, in-vitro permeation, stability and skin irritation test. In-vivo anti-inflammatory studies were performed using carrageenan induced hind paw edema method in Wistar rats. The results were compared with that of commercial nimesulide gel (CNG). From the in-vitro studies, effective permeation of nimesulide from NAE (53.04 %) was observed compared to CNG (44.72 %) at 30 min indicating better drug release from NAE. Topical application of the emulgel found no skin irritation. Stability studies proved the integrity of the formulation. The percentage of inhibition of edema was highest for the prepared NAE (67.4 % inhibition after 240 min) compared to CNG (59.6 %). From our results, it was concluded that the Aloe vera gel acts as an effective gel base to prepare nimesulide emulgel with high drug loading capacity (86.4 % drug content) compared to CNG (70.5 % drug content) with significant anti-inflammatory effect.

Improved pharmaceutical properties of surface modified bioactive plumbagin crystals

Plumbagin recrystallised by cold crystallisation technique using a variety of polar and non-polar solvents was investigated for pharmaceutical properties. Different solvents gave varying sized and shaped plumbagin. Powder X-ray diffraction, differential scanning calorimetery and fourier transform infrared spectroscopy too confirmed differing crystal habit. Platy crystals, the most significant forms, obtained from cyclohexane possessed small size (62.93 ± 3.74 μm), higher bulk density (0.108 ± 0.014 g/ml) and lower enthalpy of fusion (∆H 62.62 ± 3.67 J/g). These demonstrated approximately two-fold increase in saturation solubility (155.01 ± 3.86 μg/ml), higher Q5min (cumulative percentage dissolution in 5 min) and lower t65% (time required for 65% dissolution) owing to greater surface area. In-vivo anti-inflammatory study in Wistar rats demonstrated improvement in therapeutic efficacy of recrystallised plumbagin. In conclusion surface modification led to enhanced efficacy of plumbagin; an approach capable of improving the bioavailability and clinical efficacy of other poorly water soluble phytomedicine.

In-vitro and in-vivo anti-inflammatory activity of Syzygium alternifolium (wt) Walp

The study was aimed to evaluate the analgesic and anti-inflammatory activity (by both in-vitro and in-vivo) of both chloroform and methanol root extracts of Syzygium alternifolium(wt) Walp. Methods used for the studies are In-vitro measurement of 5-lipoxygenase inhibition and in-vivo measurement of rat paw edema and ear edema in rats, acetic acid induced writhing response and heat induced pain. Our findings clearly indicate that S. alternifolium chloroform root extract showed 5-LOX inhibition activity similar with the standard drug, Zileutin. In in-vivo anti-inflammatory studies, S. alternifolium chloroform root extract exhibited better anti-inflammatory activity in comparison to the S. alternifoliummethanolic root extract. Similar results were observed in the acetic acid induced writhing model and radiant heat induce model. Hence, it was concluded that S. alternifoliumchloroform root extract exhibited more degree of efficacy in comparison to the S. alternifolium methanolic extract and good in-vitro and in-vivo anti-inflammatory effect. Key words: Syzygium alternifolium, 5-lipoxygenase (5-LOX), analgesia, inflammation.

In-vitro and in-vivo anti-inflammatory activity of Andrographis serpyllifolia (Rottl. Ex Vahl.) Wt.

The study was aimed to evaluate the analgesic and anti-inflammatory activity (by both in-vitro and in-vivo) of both chloroform and methanol root extracts of Andrographis serpyllifolia (Rottl. Ex Vahl.) Wt. Methods used for the studies were In-vitro 5-Lipoxygenase inhibition assay and In-vivo measurement of rat paw edema and ear edema in rats, acetic acid induced writhing response and hot plate method in albino mice. Chloroform and methanolic extracts of A. serpyllifolia root have shown moderate potency in inhibiting 5-LOX and shown significant anti-inflammatory activity. Despite the IC50 values are little higher, anti-inflammatory efficacy of these extracts possibly due to other mechanisms apart of 5-LOX inhibition. However, In-vivo anti-inflammatory studies revealed that A. serpyllifolia methanolic extract has shown higher degree of efficacy when compared to the chloroform extract. In terms of analgesic activity in writhing test, methanolic extract has shown more efficacy than chloroform extract. Hence, it is important to isolate the active principles for further testing the anti-inflammatory efficacy.DOI: http://dx.doi.org/10.3329/icpj.v1i8.11250 International Current Pharmaceutical Journal 2012, 1(8): 199-204