Novel mannich-based derivative of 2-mercaptobenzimidazole (AK7): a new candidate for the treatment of inflammatory arthritis owing to its NF-κB1 inhibitory potential.

This study investigated the anti-arthritic potential of novel mannich-based derivatives of 2-mercaptobenzimidazole (AK7 and AK9) in rats. The compounds were characterized by NMR and FTIR spectroscopies and their acute anti-inflammatory effects were measured by carrageenan (CRG)-induced paw edema model. The most potent doses of AK7 and AK9 were subsequently evaluated in the complete Freund's adjuvant (CFA)-induced inflammatory arthritis model. AK7 and AK9 inhibited CRG-induced inflammation in a dose-dependent fashion and a similar reduction in CFA-induced paw inflammation was observed. Moreover, X-ray and histopathological analyses of AK7-treated animals displayed normal joint structure whereas AK9, despite of its anti-inflammatory effects, failed to protect against cartilage destruction. Interestingly, biochemical analysis revealed a better safety profile for AK7 than for AK9 and methotrexate. Both compounds suppressed mRNA levels of pro-inflammatory mediators (IRAK1, NF-κB1, TNF-α, IL1B) while only AK7 reduced the transcript levels of interstitial collagenase (MMP1). Molecular docking analysis of AK7 and AK9 with TNF-α and MMP1 also supported the experimental data. These findings clearly highlight the beneficial effects of AK7 in the prevention and/or treatment of inflammatory arthritis.

Phytochemical, anti-inflammatory, anti-nociceptive and cytotoxic basis for the use of Haloxylon stocksii.

A halophytic plant, Haloxylon stocksii, is used to treat various inflammatory disorders traditionally. The present study was carried out to investigate the phytochemical parameters, anti-inflammatory, analgesic and cytotoxic potential of the whole plant extracts of H. stocksii. The plant powder was standardized for pharmacognostic parameters. It was extracted with methanol followed by chloroform, ethyl acetate and water to prepare respective fractions. Total phenolic and flavonoid contents in the extract and fractions were estimated. The anti-inflammatory potential was determined through carrageenan-induced rat paw edema model. Centrally acting analgesic activity was assessed through the hot plate method. MTT assay was used to assess the viability of Human umbilical and human hepatocyte carcinoma cell lines upon exposure to plant extract/fractions. Chloroform fraction showed the highest phenolic while ethyl acetate exhibited a maximum flavonoids content. The plant ethyl acetate fraction exhibited highest percentage inhibition of paw edema and maximum analgesic activity at 500 mg/kg dose. The plant methanolic extract and fractions showed dose dependent cytotoxic activity. The present study concludes that the extracts of H. stocksii may be effective and safe against acute inflammatory response and pain at therapeutic concentrations.

Increasing beta cell mass to treat diabetes mellitus.

Finding a radical cure for diabetes has reached paramount importance in medicine due to the widespread prevalence of the disease. A substantial reduction in insulin-secreting beta cells is evident in diabetes. The failure of cyclin-dependent kinases (CDKs) and cyclins to access the nucleus is responsible for quiescence or senescence in human and rodent beta cells. The augmentation of beta cell proliferation is supposed to reverse diabetes. This concept has inspired the discovery of newer drugs that encourage the proliferation of beta cells. Although it is a rational step towards a cure for diabetes, the differences in biochemical pathways in rodents and human beta cells pose difficulty in promoting the proliferation of human beta cells. Primarily, it is mandatory to clearly understand the intracellular pathways involved in the proliferation of beta cells so as to pave the way for therapeutic interventions. There are several intrinsic factors that trigger the proliferation of beta cells. Furthermore, it is also obvious that the early death of beta cells due to oxidative stress-related upregulation of pro-apoptotic genes also predisposes individuals to diabetes mellitus. Polyphenols, exendin 4, histone deacetylase inhibitors, glucagon-like peptide 1, phenyl pyruvic acid glucoside, and several flavonoids reduce the early apoptosis of beta cells partly through their role in the reduction of oxidative stress. A better understanding of intracellular pathways, the identification of specific mitogens, the induction of beta cell proliferation, and the inhibition of apoptosis may help us treat diabetes mellitus through an increase in beta cell mass.

Influence of different formulation variables on the performance of transdermal drug delivery system containing tizanidine hydrochloride: in vitro and ex vivo evaluations

The present study was aimed at preparation of transdermal patches of tizanidine HCl, evaluation of the effect of polymers on in vitro release pattern of the drug, and the effect of permeation enhancers on the penetration of the drug through the rabbit skin. Various proportions of hydrophilic (HPMC) and hydrophobic (Eudragit L-100) polymers were used with PEG 400 as film-forming agent, and Span 20 or DMSO as permeation enhancer. The formulations were assessed for physicochemical characteristics and in vitro drug release studies using USP paddle over disc method in phosphate buffered saline (pH 7.4) at 32.0±1°C. On the basis of in vitro studies and physicochemical evaluations, S03-A and S04-A were selected at Eudragit : HPMC ratios of 8 : 2 and 7 : 3, respectively, for further ex vivo analysis. The effects of different concentrations of Span 20 and DMSO were evaluated on excised rabbit skin using Franz diffusion cell. Cumulative drug permeation, flux, permeability coefficient, target flux, and enhancement ratio were calculated and compared with the control formulations. Kinetic models and Tukey's multiple comparison test were applied to evaluate the drug release patterns. Formulation SB03-PE containing Eudragit L-100:HPMC (7:3) with Span 20 (15% w/w) produced the highest enhancement in drug permeation, and followed zero order kinetic model with super case-II drug release mechanism.

Genotoxic and cytotoxic potential of whole plant extracts of Kalanchoe laciniata by Ames and MTT assay.

Lack of data on safety of herbal medicines have endangered human health and life. The present study evaluated the genotoxic and mutagenic effect of Kalanchoe laciniata to access the safety and usefulness of the medicinal plant. Aqua-methanolic and n-hexane extracts of K. laciniata were evaluated for the genotoxic potential using Ames assay and cytotoxicity was evaluated using MTT assay. Ames assay was conducted using two strains of Salmonella typhimurium TA-100 and TA-102 whereas MTT assay was performed on baby hamster kidney cell line BHK-21. Aqua-methanolic extract of K.laciniata exhibited significant mutagenicity when exposed to TA-102 strain with a mutagenic index of 50.66 and 54.74 at maximum dose 150 mg/plate. The extract was also mutagenic to TA-100 strain but to a lesser extent. M.I of n-hexane extract was 12.15 and 15.51 for TA-100 and TA-102 respectively. n-hexane extract was mutagenic but little difference was observed between results of two strains. Both extracts were found to be cytotoxic with an IC50 of 321.9 and 638.5 µg/mL for aqua-methanolic and n-hexane extracts respectively. On the basis of results it was concluded that aqua-methanolic and n-hexane extracts of K.laciniata possess mutagenic and cytotoxic potential. It is suggested to explore the plant further to evaluate its safety in rodents and other species.

ANTIOXIDANT ACTIVITY OF PISTACIA KHINJUK SUPPORTED BY PHYTOCHEMICAL INVESTIGATION.

Pistacia khinjuk is one of the fifteen known species of Pistacia belonging to Anacardiaceae family. Keeping in view the possible therapeutic utility of this genus and the lack of literature on this plant, this study involves phytochemical investigation of P. khinjuk and its antioxidant activity. The phytochemical investigation was conducted on crude methanolic extract and its fractions namely, n-hexane, chloroform, n-butanol, ethyl acetate and aqueous. Total phenolic contents and flavonoids were also determined by phosphomolybdenum and ferric thiocyanate method in crude extract and its fractions. The results of phytochemical investigation indicated the presence of alkaloids, tannins, flavonoids, saponins, triterpenoids, cardiac glycosides, carbohydrates, proteins and sterols in the crude extract of P. khinjuk. Crude extract and its fractions exhibited remarkable antioxidant activity. This study showed that the crude extract and its fractions have potent antioxidant activity, among all ethyl acetate showed 1.109 ± 0.029 the highest activity. This research concluded that crude extract of P. khinjuk and its fractions contained phenolic and flavonoid compounds that show significant antioxidant activity.

EFFECT OF HYDROPHILIC AND HYDROPHOBIC POLYMER ON IN VITRO DISSOLUTION AND PERMEATION OF BISOPROLOL FUMARATE THROUGH TRANSDERMAL PATCH.

A matrix transdermal patch of bisoprolol fumarate was formulated with different concentrations of Eudragit RS 100 and Methocel E5 with PEG 400 as plasticizer by solvent evaporation technique. Tween 80 was added to the optimized patch to evaluate the effect of permeation enhancer at different concentration through the excised rabbit's skin. The patches were analyzed for weight variation, drug content, swelling index, erosion studies, moisture content, moisture uptake, water vapor transmission rate (WVTR) and water vapor permeability (WVP). In vitro dissolution test was carried out in USP dissolution apparatus V to select the optimized formulation. In vitr skin permeation studies were done in Franz diffusion cell using rabbit skin as a model membrane. The cumulative drug release and flux were determined to compare the result of test patches with a control patch. The greatest enhancement ratio (ER) was obtained in F03-PE with 30% Tween 80. F03-PE seemed to follow zero order kinetics with super case II mechanism of drug release. Statistical ANOVA suggested that there was a significant difference in formulations, steady flux and cumulative permeation rate at different Tween 80 concentrations.

Genotoxic and cytotoxic action potential of Terminalia citrina, a medicinal plant of ethnopharmacological significance.

Most herbal medicines utilized in complementary and alternative medicine lack safety evaluation setting our lives under unwarranted risks. Present study comprised of genotoxic and cytotoxic appraisal of Terminalia citrina fruits which are used as a folklore medicine for treatment of various ailments. Aqueous and ethanolic extracts of T. citrina fruit extracts were evaluated for the presence of different phytochemicals. Genotoxic potential of both the extract of T. citrina was assessed through Ames reverse mutagenicity assay in Salmonella TA 100 and 102 strains. Cytotoxic potential of T. citrina was determined in baby hamster kidney cell line (BHK-21). Statistical analysis was carried out by ANOVA following post hoc test. Phytochemical analysis showed the presence of alkaloids, carbohydrates, phenolic compounds, tannins, catechins and saponins. It was revealed that both the extracts of T. citrina exhibited significant mutagenicity in tester strains. Ethanolic extract showed higher mutagenicity in TA 100 strain, whereas aqueous extract of T. citrina exhibited higher mutagenicity in TA 102 strain than TA 100. Both the extracts of T. citrina showed dose-dependent mutagenicity. Fifty percent cell viability was exhibited by 260 and 545 µg/mL of ethanolic and aqueous extracts respectively. This study concludes that the ethanolic and aqueous fruit extracts of T. citrina may not be safe owing to their mutagenic and cytotoxic potential and it necessitates further investigation regarding its safety evaluation.

The formulation of flurbiprofen loaded microspheres using hydroxypropylmethycellulose and ethylcellulose.

OBJECTIVES: The aim of the present work was to formulate flurbiprofen (FLB) loaded microspheres of hydroxypropylmethycellulose and ethylcellulose polymers to study the effect of different proportions of the polymer mixture on the release behavior of the drug. MATERIAL AND METHODS: A series of microspheres were prepared using tween-80 as a surfactant. The prepared microspheres were evaluated for entrapment efficiency (%) and percentage recovery. Drug release was performed in USP phosphate buffers of pH 1.2 and 6.8. Drug release data were plotted in various kinetic models, including zero-order, first-order, Higuchi and Korsmeyer-Peppas models to investigate the optimum composition suitable for sustained drug delivery. RESULTS: A significant difference in drug release kinetics was observed by varying the composition of hydroxypropylmethycellulose/ethylcellulose. As the ratio of EC/HPMC was increased, the release rate of flurbiprofen decreased. CONCLUSIONS: This study demonstrated the potential of polymer combinations in the formulation of microspheres for water-insoluble drugs utilizing HPMC and EC as release retardant materials, using a simple solvent evaporation microencapsulation technique. It was observed that various physico-chemical properties of the microspheres varied according to the change in polymer concentrations used in the formulations.