Md. Khalid Anwer

Diosmin downregulates the expression of T cell receptors, pro-inflammatory cytokines and NF-κB activation against LPS-induced acute lung injury in mice.

Diosmin, a natural flavonoid glycoside present abundantly in the pericarp of various citrus fruits. Because of its anti-inflammatory and antioxidant properties, it can be used in many diseases. In this study, we investigated the possible protective mechanisms of the diosmin on LPS-induced lung injury through inhibition of T cell receptors, pro-inflammatory cytokines and NF-κB activation. Animals were pretreated with diosmin (50 and 100mg/kg, p.o.) for seven days prior to lipopolysaccharides (LPS) treatment. LPS administration increased neutrophils, monocytes, lymphocytes, total leukocyte count (TLC) and platelets which were decreased by diosmin. We observed that mice exposed to LPS showed increased malondialdehyde level and MPO activity whereas marked decrease in glutathione content. These changes were significantly reversed by treatment with diosmin in a dose dependent manner. Diosmin treatment showed a substantial reduction in T cell (CD4(+) and CD8(+)) receptors and pro-inflammatory (IL-2(+) and IL-17(+)) cytokines in whole blood. In addition, RT-PCR analysis revealed increased mRNA expression of IL-6, IL-17, TNF-α, and NF-κB in the LPS group, while reduced by treatment with diosmin. Western blot analysis confirmed the increased protein expression of IL-1β, TNF-α and NF-κB p65 in the LPS group and treatment of animals with diosmin reversed these effects. The levels of cytoplasmic p-IκB-α and p-NF-κB p65 expression also were mitigated by diosmin. The histological examinations revealed protective effect of diosmin while LPS group aggravated lung injury. These results support the potential for diosmin to be investigated as a potential agent for the treatment of lung injury and inflammatory diseases.

Enhanced oral bioavailability of insulin-loaded solid lipid nanoparticles: pharmacokinetic bioavailability of insulin-loaded solid lipid nanoparticles in diabetic rats.

Abstract Objective: Insulin is a hormone used in the treatment of diabetes mellitus. Multiple injections of insulin every day may causes pain, allergic reactions at injection site, which lead to low patient compliance. The aim of this work was to develop and evaluate an efficient solid lipid nanoparticle (SLN) carrier for oral delivery of insulin. Methods: SLNs were prepared by double emulsion solvent evaporation (w/o/w) technique, employing glyceryltrimyristate (Dynasan 114) as lipid phase and soy lecithin and polyvinyl alcohol as primary and secondary emulsifier, respectively, and evaluated in vitro for particle size, polydispersity index (PDI) and drug entrapment. Results: Among the eight different developed formulae (F1–F8), F7 showed an average particle size (99 nm), PDI (0.021), high entrapment of drug (56.5%). The optimized formulation (F7) was further evaluated by FT-IR, DSC, XRD, in vitro release, permeation, stability, bioavailability and pharmacological studies. Insulin-loaded SLNs showed better protection from gastrointestinal environment as evident from the relative bioavailability, which was enhanced five times as compared to the insulin solution. A significant enhancement of relative bioavailability of insulin was observed, i.e. approximately five times of pure insulin solution when loaded in SLN (8.26% versus 1.7% only).

Development and evaluation of PLGA polymer based nanoparticles of quercetin

Quercetin is the most abundant antioxidant found in the human diet. Low aqueous solubility of quercetin limits its bioavailability and hence therapeutic effects. Therefore, the aim of the present study was to develop a poly lactide-co-glycolic acid (PLGA) polymer based nanoparticles of quercetin with a view to improve its aqueous solubility and examine the effect on its antioxidant and diuretic properties. Nanoparticles of quercetin were developed by single emulsion-solvent evaporation technique and evaluated in vitro for differential scanning calorimetry (DSC), Fourier transforms infra-red (FTIR) spectroscopy, particle size, polydispersity index and drug entrapment efficiency. Among the five different formulations (F1, F2, F3, F4 and F5), F2 and F3 were optimized with an average particle size of 189nm and 186nm and high entrapment values of 86.48%, 83.71%, respectively. SEM images of confirmed that prepared nanoparticles were spherical in shape with a smooth surface. In vitro release and anti-oxidant activity confirmed significant results. Furthermore, its in vivo diuretic activity was much better as compared to pure quercetin. The overall results suggest that PLGA polymer based nanoparticle could be a potential option for quercetin delivery.

Development of Curcumin loaded chitosan polymer based nanoemulsion gel: In vitro, ex vivo evaluation and in vivo wound healing studies

In the present study, various nanoemulsions were prepared using Labrafac PG+Triacetin as oil, Tween 80 as a surfactant and polyethylene glycol (PEG 400) as a co-surfactant. The developed nanoemulsions (NE1-NE5) were evaluated for physicochemical characterizations and ex-vivo for skin permeation and deposition studies. The highest skin deposition was observed for NE2 with 46.07% deposition amongst all developed nanoemulsions (NE1-NE5). Optimized nanoemulsion (NE2) had vesicle size of 84.032±0.023nm, viscosity 78.23±22.2 cps, refractive index 1.404. Nanoemulsion gel were developed by incorporation of optimized nanoemulsion (NE2) into 1-3% chitosan and characterized by physical evaluation and rheological studies. Chitosan gel (2%) was found to be suitable for gelation of nanoemulsion based on its consistency, feel and ease of spreadability. The flux of nanoemulsion gel was found 68.88μg/cm2/h as compared to NE2 (76.05μg/cm2/h) is significantly lower suggesting limited skin permeation of curcumin form gel. However, the retained amount of curcumin on skin by gel formulation (980.75±88μg) is significantly higher than NE2 (771.25±67μg). Enhanced skin permeation of NE2 (46.07%) was observed when compared to nanoemulsion gel (31.25%) and plain gel (11.47%). The outcome of this study evidently points out the potential of curcumin entrapped nanoemulsion gel in wound healing.

Wound healing effects of nanoemulsion containing clove essential oil

Abstract The aim of this study was to investigate the wound healing effects of clove oil (CO) via its encapsulation into nanoemulsion. Optimized nanoemulsion (droplet size of 29.10 nm) was selected for wound healing investigation, collagen determination, and histopathological examination in rats. Optimized nanoemulsion presented significant would healing effects in rats as compared to pure CO. Nanoemulsion also presented significant enhancement in leucine content (0.61 mg/g) as compared to pure CO (0.50 mg/g) and negative control (0.31 mg/g). Histopathology of nanoemulsion treated rats showed no signs of inflammatory cells. These results suggested that nanoemulsion of CO was safe and nontoxic.

Development and evaluation of olmesartan medoxomil loaded PLGA nanoparticles

The purpose of the present study was to develop olmesartan medoxomil (OLM) loaded poly (d,l-lactide-co-glycolide) (PLGA) nanoparticles by nanoprecipitation technique. Olmesartan medoxomil nanoparticles (F1–F3) were prepared using PLGA as polymer and Pluronic acid 127 as a surfactant. Developed PLGA nanoparticles were evaluated for particle size, polydispersity index (PI), differential scanning calorimetry (DSC), Fourier transforms infrared (FTIR) spectroscopy, powder X-ray diffraction (XRD), scanning electron microscopy (SEM), drug entrapment efficiency (EE%) and in vitro drug release. Optimised PLGA nanoparticle (F3) had particle size of 97.8 ± 6.3 nm, EE of 58.33% and drug release of 37.1% after 24 h of study. SEM images confirmed that developed nanoparticles were spherical in shape with a smooth surface. DSC, FTIR, and XRD spectra indicated formation of PLGA nanoparticles. The results suggest that PLGA polymer-based nanoparticle could be a potential option for sustained drug delivery of OLM.

Quantitative estimation of hesperidin by HPTLC in different varieties of citrus peels

OBJECTIVE To develop a simple, selective, sensitive and accurate high-performance thin layer chromatography (HPTLC) method to determine the quantity of hesperidin in different varieties of citrus fruits. METHODS The method was carried out in aluminum-backed silica gel 60 F254 plates with ethyl acetate-methanol-water 15:3:2 (%, v/v) as mobile phase. RESULTS A compact band was obtained for hesperidin at Rf value of (0.40±0.04). The calibration plot was linear in the range of 100-800 ng/spot of hesperidin and the correlation coefficient of 0.998 6 was indicative of good linear dependence of peak area on concentration. Limit of detection (8.87 ng/spot), limit of quantification (23.21 ng/spot), accuracy (less than 2%) and recovery (ranging from 98.55-99.38) were found satisfactory. CONCLUSIONS The method developed can be used for routine analysis of hesperidin in crude drug as well as in herbal and pharmaceutical dosage form containing citrus fruits as an ingredient.

QUALITATIVE AND QUANTITATIVE ANALYSIS OF KHELLIN IN AMMI VISNAGA FRUITS AND PHARMACEUTICAL PREPARATIONS USING HPTLC AND HPLC

An attempt was made to develop and validate a simple and accurate HPTLC method for the analysis of khellin in Ammi visnaga fruits and some pharmaceutical preparations. HPLC method was developed in order to verify the results of HPTLC analyses. HPTLC method was developed using chloroform-acetone 9:1 (%, v/v) as mobile phase. The analysis was performed on 10 × 20 cm aluminum-backed plates coated with 0.2-mm layers of silica gel 60 F254 (E-Merck, Germany). Camag TLC Scanner III was used for the UV densitometric scanning. RP-HPLC method was developed using a mobile phase composed of methanol-water 60:40 (%, v/v) and C18-ODS-Hypersil column in isocratic mode. Analyses were done in UV absorbance mode at 248 nm for both. The used HPTLC system was found to give sharp, symmetrical, and well-resolved peak at Rf value of 0.29 ± 0.04 and linearity in the range 50–300 ng/spot (r2 = 0.9939). On the other hand, the peak of khellin was well-resolved at Rt 4.73 min and linearity in the range of 100–1000 ng/mL (r2 = 0.9960) was used in the HPLC method. These results indicated that proposed HPTLC and HPLC methods could be successfully applied for quality control and routine analysis of khellin in commercial formulations.

High throughput μ-SPE based elution coupled with UPLC–MS/MS for determination of eluxadoline in plasma sample: Application in pharmacokinetic characterization of PLGA nanoparticle formulations in rats

Graphical abstract Figure. No caption available. HighlightsA UPLC–MS/MS assay for analysis of eluxadoline in plasma.Use of &mgr;‐SPE technique for sample extraction.Assay applied in bioavailability study in rats. ABSTRACT Eluxadoline is a novel &mgr;‐ and &kgr;‐opioid receptor (OR) agonist and &dgr;‐OR antagonist, recently approved as a first line therapy for the treatment of irritable bowel syndrome. Due to abuse potential, poor bioavailability and high intersubject variability, a sensitive and reliable assay is prerequisite for its determination in biological samples. This work first time report the development and validation of UPLC–MS/MS assay for determination of eluxadoline in rat plasma sample using risperidone as an internal standard (IS). A high‐throughput 96‐well plate format &mgr;‐SPE technique was used for plasma sample extraction. The extracted samples were separated on Acquity BEH™ C18 column (100 × 2.1 mm, 1.7 &mgr;m) using mobile phase elution of acetonitrile: 20 mM ammonium acetate (80:20, v/v) at a flow rate of 0.3 mL min−1. The precursor to product ion transition of m/z 570.16 → 118.12 (qualifier), 570.16 → 171.08 (quantifier) for eluxadoline, and m/z 411.18 → 191.07 for IS were used for MRM monitoring. The calibration curves were linear in concentration range of 0.15–50 ngmL−1 with LOD and LOQ of 0.07 and 0.15 ngmL−1, respectively. The validation results satisfied the criteria of USFDA and SWGTOX guidelines and were within the acceptable limit. Finally, the method was successfully applied in bioavailability enhancement study of the newly developed PLGA nanoparticles and Eudragit coated PLGA nanoparticles of eluxadoline in rats.

Preparation, Evaluation and Bioavailability Studies of Eudragit Coated PLGA Nanoparticles for Sustained Release of Eluxadoline for the Treatment of Irritable Bowel Syndrome

Eluxadoline is a newly approved orally administered drug used for the treatment of Irritable Bowel Syndrome with Diarrhea. It is reported as a poorly water-soluble drug due to which its dissolution rate and oral bioavailability are very poor. In this work, various plain PLGA nanoparticles (NPs) (F1–F4) were prepared and optimized based on particle size, PDI, zeta potential and percent drug entrapment efficiency (EE). The developed plain NPs (F1–F4) showed average particle size ranging from 260.19 to 279.76 nm with smooth surface and EE of 17.83–56.29%. The optimized plain NPs (F3) had particle size of 273.76 ± 7.25 nm with a low PDI value 0.327, zeta potential - 30.63 ± 2.47 mV and % EE of 56.29 ± 2.56%. The optimized F3 NPs was further submitted for enteric coating using Eudragit S100 polymer and evaluated in terms of particles characterization, in vitro release and pharmacokinetic studies in rats. The bioavailability of plain and coated nanaoparticles were enhanced by 6.8- and 18.5-fold, respectively, compared to normal suspension. These results revealed that the developed coated NPs could be used for its oral delivery for an effective treatment of Irritable Bowel Syndrome with Diarrhea.