ANTIBACTERIAL ACTIVITY OF CHLOROGENIC ACID PHYTOVESICLES AGAINST RESISTANT BACTERIA: DEVELOPMENT, OPTIMIZATION AND EVALUATION

Objective: To investigate the in vitro antibacterial activity of a naturally occurring polyphenol chlorogenic acid (CGA) and compares it with formulated chlorogenic acid phytovesicles against 4 different bacterial strains; two gram positive [Staphylococcous aureus and Bacillus subtilis] and two gram negative strains [Klebsiella pneumonia and Escherichia coli]. Methods: CGA phytovesicles were developed and optimized using central composite design to improvise CGA’s physicochemical properties. Bactericidal activity was evaluated using agar diffusion, minimum inhibitory concentration (MIC) and time kill assay. The effect of pH and temperature on the antimicrobial activity was determined. Results: The optimized CGA phytovesicles showed entrapment of 96.89% with 30 times better lipophilic solubility than the plain drug. The inhibition zone sizes for CGA phytovesicle ranged from 17-25 mm as compared to 15-20 mm of plain CGA while the MIC values ranged 200-250 µg/ml as compared to 500-550 µg/ml of plain CGA. CGA phytovesicles exhibited a strong bactericidal effect at MIC with a log reduction in the range of 0.90-2.04 in Colony forming units (CFUs) at 24h for different strains as compared to 1.38-2.17 of plain CGA. Furthermore, the antibacterial effect was found to augment with increasing temperature but decreased with alkaline pH. Conclusion: Results strongly supports the hypothesis of potential use of CGA phytovesicles as a mode of drug delivery for its antibacterial use against different resistant bacteria

DEVELOPMENT OF MULTIPARTICULATE FORMULATION AND EVALUATION OF COLON TARGETED DRUG DELIVERY SYSTEM OF KETOPROFEN: IN VIVO STUDY WITH INDUCED COLITIS MODEL IN RATS AND GAMMA SCINTIGRAPHY

Objectives: To develop a multiparticulate containing chitosan and guar gum for the treatment of ulcerative colitis.Methods: The formulation of multiparticulate was done by extrusion spheronization method using Eudragit L-100 and Eudragit S-100 as a coatingsolution and Ketoprofen as a model drug.Results: Preliminary trial batches were previously assessed for physicochemical characterization, in vitro release, ex vivo mucoadhesion study,swelling studies, and in vivo evaluation and showed that the formulations appeared to be a good candidate to deliver the drug to the colon. BoxBehnkendesign wasused tostatisticallyoptimize the formulationparametersandevaluatethe main effects,interactioneffects,and quadraticeffectsof theprocessparametersofentericcoatedmultiparticulateon drugpolymerratioand coat composition. Inthis work,the effectivenessofoptimizedbatch(K10) inthe treatmentof inflammatoryboweldisease wasevaluated.Experimentally,colitis wasinduced byrectalinstillationof2,4,6,trinitrobenzenesulfonicacidintomaleWistar rats.The histologicalevaluationsweredoneasinflammatoryindices.Invivogammascintigraphystudiesof multiparticulatewithout drug demonstratedegradationof multiparticulatewhenevertheyreachthe colon.Conclusion: Results of studies like Gamma Scintigraphy and Histological study of optimized formulation (K10) clearly indicate that there is a greatpotential in the delivery of Ketoprofen to the colonic region. The animals treated with Ketoprofen (K10) formulation had an improvement in pathologyand may be useful for the treatment of inflammatory bowel disease.Keywords: Chitosan, Guar gum, Ketoprofen, Gamma scintigraphy, Histology, Ulcerative colitis, Box-Behnken design

NOVEL UMBELLIFERONE PHYTOSOMES: DEVELOPMENT AND OPTIMIZATION USING EXPERIMENTAL DESIGN APPROACH AND EVALUATION OF PHOTO-PROTECTIVE AND ANTIOXIDANT ACTIVITY

Objective: The objective of this study was to develop a novel formulation (Phytosome) of umbelliferone with phospholipid for improved permeability, solubility and hence better pharmacological action.Methods: The phytosomal complex was prepared by using solvent evaporation method and optimised by applying the Box-Behnken design on the basis of complexation rate and partition coefficient. The formation of phytosomes was confirmed by FTIR, DSC, SEM, XRD, HPTLC and NMR by comparing the results of the complex with the drug. The photoprotective potential of complex against UV-exposure was evaluated in rats and compared with the drug by incorporating it in a gel and estimating the antioxidant enzymes in skin namely reduced glutathione, superoxide dismutase, lipid peroxidation and catalase.Results: The crystalline drug was completely converted to amorphous complex. The complex showed a good practical yield, drug content and particle size was in the range. The solubility of the complex was determined by partition coefficient method and was found to be better than the drug. The ex vivo and in-vitro permeation of the complex showed improved permeation for complex than the drug. The in vitro antioxidant activity of complex was evaluated by DPPH and ferrozine antioxidant assay and was better than the drug. The photoprotective action of the complex was found to be better than drug on the basis of the content of antioxidant enzymes estimated in the skin.Conclusion: The phytosomal complex was found to show better solubility in the water phase and oil phase, better permeation, better antioxidant activity and a better photo-protective activity when compared to umbelliferone

Formulation, Optimization and Evaluation of Self Emulsifying Immediate Release Tablet of Nebivolol HCl using 32 Factorial Design

Nebivolol Hydrochloride (NEB) is a lipophilic molecule with low solubility in GI fluid, and high metabolism which leads to its low oral bioavailability 12%. The aim of the present investigation was to develop immediate release self emulsifying tablet (IR-SET) as solid SMEDDS to enhance the solubility and permeability of the drug. Solubility study, pseudo-ternary phase diagrams and 32 factorial design were used to select the components of the system and optimize the composition of liquid SMEDDS. Optimal L-SMEDDS contains Kollisolv GTA, Tween 80 and Propylene glycol as oil, surfactant and co-surfactant, respectively in the ratio of 20:26.66:53.34 % w/w, formulates L-SMEDDS with droplet size (55.98 nm), PDI (0.37), emulsification time (16±1.52 sec) and drug content (97.43±0.30 %).  The liquid SMEDDS were adsorbed onto Neusilin US2 by adsorbtion technique to form S-SMEDDS. DSC and SEM studies suggested that NEB in the S-SMEDDS may be present in the molecular dispersed state and was sufficiently adsorbed onto solid carrier, respectively. S-SMEDDS was compressed into IR-SET by direct compression method and composition of IR-SET was optimized using 32 factorial design. Optimal IR-SET showed disintegration time (92 + 0.57 sec), droplet size (68.57 nm), PDI (0.34) and drug content (96.33±0.15 %). In vitro dissolution studies and ex vivo diffusion studies in rat stomach suggested that SMEDDS played an important role in solubility and permeability enhancing effect. Accelerated stability studies indicated that formulation were stable. Our results illustrated the increase in solubility and permeability of drug from IR-SET

DEVELOPMENT OF MULTIPARTICULATE FORMULATION AND EVALUATION OF COLON TARGETED DRUG DELIVERY SYSTEM OF CIPROFLOXACIN: IN VIVO STUDY WITH INDUCED COLITIS MODEL IN RATS

Objective: The objective of this research work was to develop a multiparticulate containing chitosan and guar gum for the treatment of ulcerative colitis.Method: Method for the formulation of multiparticulate was done by extrusion-spheronization method using Eudragit L-100 and Eudragit S-100 as a coating solution and ciprofloxacin as a model drug.Result: Result from preliminary trial batches was previously assessed for physicochemical characterization, in vitro release, ex vivo mucoadhesion study, swelling studies, and in vivo evaluation and showed that the formulations appeared to be a good candidate to deliver the drug to the colon. Box-Behnken design was used to statistically optimize the formulation parameters and evaluate the main effects, interaction effects, and quadratic effects of the process parameters of enteric-coated multiparticulate on drug polymer ratio and coat composition. In this work, the effectiveness of optimized batch (C4) in the treatment of inflammatory bowel disease was evaluated. Experimentally, colitis was induced by rectal instillation of 2, 4, 6, trinitrobenzene sulfonic acid into male Wistar rats. The histological evaluations were done as inflammatory indices. In vivo gamma scintigraphy studies of multiparticulate without a drug demonstrate degradation of multiparticulate whenever they reach colon.Conclusion: The conclusion from results of studies such as gamma scintigraphy and histological study of optimized formulation (C4) clearly indicates that there is a great potential in delivery of ciprofloxacin to the colonic region. The animals treated with ciprofloxacin (C4) formulation had an improvement in pathology and may be useful for the treatment of inflammatory bowel disease.Keywords: Chitosan, Guar gum, Ciprofloxacin, Gamma scintigraphy, Histology, Ulcerative colitis, Box-Behnken design

Transdermal iontophoretic delivery of timolol maleate

Transdermal iontophoresis would be a promising method for the systemic delivery of water soluble and ionic drugs of relatively high molecular size, including peptides. The objective of the present study was to investigate the effect of biological variable such as guinea pig and human cadaver skin and other variables like drug concentration, current density on the transdermal iontophoretic transport of timolol maleate. The permeation profile of drug using solution and gel formulation was studied and compared. For better bioavailability, better patient compliance, and enhanced delivery, an iontophoretic drug delivery system of a timolol maleate matrix gel was formulated using Carbopol 974P. The study was conducted using silver-silver chloride electrodes across the guinea pig and human cadaver skin. Viscosity measurements and flux calculations indicated the suitability of the Carbopol 974P gel for transdermal iontophoretic delivery of timolol maleate. Anodal iontophoresis with silver-silver chloride electrode significantly increased the timolol maleate skin permeation as compared with the passive permeation study. The amount of timolol maleate transported during iontophoresis was significantly different among the different skins. However, iontophoretic gel formulations provided required flux of drug through human cadaver skin.A iontoforese transdérmica seria um método promissor para a liberação sistêmica de fármacos solúveis em água e iônicos de relativamente elevado tamanho molecular, incluindo peptídeos. O objetivo do presente estudo foi investigar o efeito da variável biológica, tais como cobaia e pele de cadáver humano, e outras variáveis como concentração do fármaco, densidade de corrente sobre o transporte transdérmico iontoforético de maleato de timolol. Comparou-se o perfil de permeação do fármaco usando a formulação de solução e de gel. Para melhor biodisponibilidade, melhor adesão do paciente e liberação aprimorada, formulou-se sistema de liberação iontoforética gel de maleato de timolol usando Carbopol 974P. O estudo foi conduzido usando eletrodos de prata-cloreto de prata na cobaia e na pele de cadáver humano. Medidas de viscosidade e de fluxo indicaram a adequação do gel Carbopol 974 P para liberação iontoforética transdérmica do maleato de timolol. A iontoforese anódica com eletrodo de prata-cloreto de prata aumentou significativamente a permeação dérmica do maleato de timolol, comparativamente à permeação passiva. A quantidade de maleato de timolol transportado durante a iontoforese foi significativamente diferente entre as diferentes peles . No entanto, as formulações iontoforéticas de gel forneceram o fluxo necessário do fármaco através da pele de cadáver humano

Mucoadhesive carbamazepine gel for in situ olfactory delivery

Purpose: To formulate mucoadhesive carbamazepine gel for delivery to the brain via the olfactory mucosa. Methods: Carbamazepine transfersomes were formulated using Lipoid S 100 and sodium cholate. The transfersomes were evaluated for entrapment efficiency, in vitro release transmission electron microscopy, zeta potential, polydispersity index. The transfersomes were then incorporated into gellan gum gel, and the in situ gel formulation was evaluated for drug content, gel strength, in vitro release and mucoadhesive force. Transfersomes were also evaluated for bioanalytical study in rats. Result: TEM analysis showed good regular spheres. The negative zeta potential ensures resistance to aggregation. The gel strength of the formulations was in the range of 0.6 to 7.4 g. In vitro diffusion study of transfersomal gel showed Fickian diffusion mechanism. Formulation F6 was optimized depending for gel strength (6.4 g) , drug content (99.47 ± 0.25 %), and good mucoadhesive force (50.24 ± 0.76 dyne/cm2). Bioanalytical study of F6 showed increased drug concentration in brain. Conclusion: Mucoadhesive carbanmazepine gel can be used effectively to achieve increased concentration of drug in the brain via olfactory mucosal rout

Dental Patient’s Knowledge and Awareness Regarding Effects of Smoking on Oral Health among Smokers and Nonsmokers: A Comparative Study

Aims and objectives: The aim of the present study is to examine the difference in dental patient’s knowledge and awareness regarding effects of smoking on oral health betweensmokers and nonsmokers. Materials and methods: A cross-sectional questionnaire-based survey was carried out among 199 dental patients from Sri Aurobindo Institute of Dental Science, Indore. A self-prepared questionnaire was used to assess the awareness regarding the effects of smoking on oral health. Chi-square test and multiple linear logistic regression model was applied using statistical package for social sciences (SPSS) software. Results: The prevalence of smoking was 22.6%. Fewer smokers and tobacco users than nonusers thought that oral health and smoking are related (28.0 vs 63.3%, p = 0.009). Multiple linear logistic regression was used to assess the association of each variable with awareness, which showed that smokers are less aware of the oral health effects of smoking than the nonsmokers. Conclusion: Smoking and tobacco using subjects are significantly less aware of the oral health effects of smoking as compared to nonsmokers. Comparative studies in other populations may be warranted to ascertain the validity of these results

Experimental design approach for development of novel microemulsion system and immediate release self microemulsifying tablet of nebivolol HCl

The objective of this study was to determine specific combination of pharmaceutical excipients that lead to formulation of efficient nebivolol hydrochloride SMEDDS and its subsequent formulation into IR-SET (Immediate release- Self emulsifying tablet) which will enhance its solubility and dissolution. Solubility and Pseudo-ternary phase studies were carried out to identify the excipients showing highest solubility and to identify the zone of microemulsion with selected ingredients. Liquid-SMEDDS (L‑SMEDDS) were optimized for Concentration of oil(X1) and Smix(X2) and formulated using a combination of Kollisolv GTA as oil, Tween 80 as surfactant and propylene glycol as co-surfactant which gave smaller droplet size(Y1) 55.98nm , Emulsification time (Y2) 16±1.5 s,% transmittance (Y3) 99.94±0.47%. Neusilin US2 was used as solid carrier for solidification of L-SMEDDS in to Solid-SMEDDS (S-SMEDDS) by adsorption technique. IR-SET of nebivolol were formulated with S-SMEDDS and optimized for the concentration of binder (X1) (PVP K30) and superdisintegrant (X2) (KOLLIDON CL) which showed low Disintegration time (Y1) (92±0.5s) and low Friability(Y2)(0.424±0.03%). Also the DSC and XRD data revealed the molecular state of the drug in S-SMEDDS. The extent of in-vivo drug release and ex-vivo diffusion values from L-SMEDDS and IR-SET was much higher than pure drug and marketed tablet. In conclusion, the results showed potential of SMEDDS to improve solubility and thus the bioavailability

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF DABIGATRAN ETEXILATE RELATED SUBSTANCE IN PHARMACEUTICAL DOSAGE FORM BY REVERSE‑PHASE – HIGH‑PERFORMANCE LIQUID CHROMATOGRAPHY

Objective: The objective of the study was to develop and validate new, simple, and selective reverse-phase–high-performance liquid chromatography (RP-HPLC) method for the quantitative determination of Dabigatran Etexilate (DE) and its impurities in pharmaceutical dosage form as per the International Conference on Harmonization guidelines.Method: Chromatographic analysis was performed on Princeton SPHER-l00 C18 (250 × 4.6 mm, 5 μm) HPLC column, maintained at 50°C column temperatures, 6°C sample tray temperature, and detection monitored at 225 nm. The mobile phase consisted of acetonitrile:phosphate buffer (pH 2.5) (33:67 V/V). The flow rate was maintained at 1.0 ml/min.Results: The system suitability results indicate good performance of the system. Specificity study indicates that there is no interference of placebo and blank. The percentage relative standard deviation (RSD) of six preparations for known and unknown impurity in the sample solution is found below 10%; hence, the method is precise. The calibration curve for DE (unknown impurity), Impurity A was linear from 0.38 to 4.5 μg/ml (correlation coefficients [r2] for unknown Impurity [DE] and Impurity A are 0.996 and 0.999, respectively). The calibration curve for Impurity B and Impurity E was linear from 0.38 to 9.00 μg/ml (r2 for Impurity B and Impurity E are 0.999 and 0.999, respectively); hence, the method is linear. Accuracy for DE (unknown Impurity), Impurity A, Impurity B, and Impurity E are found within 80%–120%; hence, the method is accurate. The percentage RSD for a standard solution is found below 5% up to 24 h, and percentage impurity change in the sample solution is found below 0.1% up to 18 h; hence, standard solution is stable up to 24 h, and sample solution is stable up to 18 h.Conclusion: The developed method is new, simple, adequate, specific, precise, linear, and accurate for the determination of DE and its impurities in pharmaceutical dosage forms