ANTI-ALZHEIMER ACTIVITY OF TACRINE LOADED METHOXYPOLY (ETHYLENE GLYCOL) POLY (CAPROLACTONE) NANOPARTICLES

Objective: The objective of the study was to study the in vitro cytotoxic effects of amyloid-β (Aβ) in SH-SY5Y cells and in vivo therapeutic efficacy of drug-loaded nanoparticles (TMPCN-3) in mice.Methods: In vitro cytotoxic effects of Aβ in SH-SY5Y cells were carried out using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay; cellular uptake study was carried using a laser scanning fluorescence microscope. In vivo therapeutic efficacy of drug loaded nanoparticles, optimized formulation (TMPCN-3) in mice. Animals (n=18) were divided into three different groups each consisting of six animals in each group.Results: Treatment with Aβ-induced cytotoxicity showed a 54% decrease in cell viability. Drug-treated with TMPCN-3 showed greater intracellular accumulation. In the invasive study TMPCN-3 nanoparticles, the SH-SY5Y cells (73%) were decreased when compared to the control cells (91%). In vivo therapeutic efficacy of drug-loaded nanoparticles was studied in mice, Morris water maze task shows polymeric nanoformulations indicates critical change execution (expanded memory maintenance) when contrasted with control, step down condition maintains a considered space from response created by the memory change and polymeric nanoformulations, and after that taken by tacrine and control the quantity of errors are higher. The nanoformulations distributed massive decreasing in acetylcholine esterase level brain homogenate.Conclusion: Tacrine-loaded methoxypoly(ethylene glycol) poly(caprolactone) reveals that the in vitro cell line believes emphasized the reduced risk of tacrine loaded nanoparticles as there is a significant decrease half maximal inhibitory concentration and higher convention of drug in the glial cells and by in vivo reflects was confirmed that nanoformulation indicated improved memory

PREPARATION AND CHARACTERIZATION OF GEMCITABINE LOADED MPEG-PCL POLYMERIC NANOPARTICLES FOR IMPROVED TRANSPORTATION ACROSS BLOOD BRAIN BARRIER

Objective: To prepare Gemcitabine (GCB) loaded Methoxy Polyethylene Glycol-Poly (Caprolactone), (MPEG-PCL) nanoformulations and to carry out the physicochemical characterisation with a primary objective to enhance the transport and penetration of drug across the blood-brain barrier (BBB).Methods: Gemcitabine loaded MPEG-PCL nanoparticles were formulated by using modified nanoprecipitation method. Nanoformulations were prepared by varying drug: polymer ratio. The prepared nanoparticles (NP) were evaluated for particle size, zeta potential, entrapment efficiency, drug content and in-vitro drug release studies. The in vitro cytotoxicity of drug-loaded NPs was evaluated in U-87 MG cells. Results: The prepared nanoformulations indicated a significant increase in particle size with increase in the polymeric concentration. GCB loaded MPEG-PCL nanoformulation (GCBNP 3) exhibited a particle size of 223±1.4 nm. DSC thermo grams indicated that GCB was dispersed as an amorphous state in MPEG NPs. SEM, TEM, and AFM studies indicated that the NPs were spherical, smooth surface without any cracks or pinholes. In vitro studies showed the GCBNP 3 shows an initial burst release followed by a more gradual and sustained-release phase (maximum drug release). The cytotoxicity of GCB loaded MPEG-PCL nanoformulations showed reduction in the IC50 value (4.1 µg). Apoptosis detection assay with Hoechst 33342 dye was carried out and observed an increase in fluorescence in the apoptotic cells. By invasive studies, the GCB loaded MPEG-PCL nanoformulation inhibits the cell migration significantly when compared with the pure drug.Conclusion: The GCB loaded MPEG-PCL nano particles indicated improved cytotoxic activity with minimal concentrations compared with the pure drug in U-87 MG glial cells. Hence, it can be concluded that GCB loaded MPEG-PCL nanoformulation can serve as a potential drug delivery tool for the treatment of brain tumours.Â

FORMULATION AND IN-VITRO EVALUATION OF EFAVIRENZ LIQUISOLID COMPACTS

Objective: The present research is aimed to enhance the dissolution rate of Efavirenz using liquisolid compact technology.Methods: About 16 different formulations were developed using factorial design with carriers (Neusilin and Fugicalin), binder (PVP K-30) and vehicle (polyethylene glycol 300) as independent variables and aerosil 200 is used as coating material. The In-vitro drug release from the LSC has used a dependent variable. The empirical method by Spireas and Bolton was applied to calculate the amounts of carrier and coating materials and obtained the improved flow characteristics and hardness by changing the proportion of carrier and coating materials.Results: A 23 factorial design is used and developed LSC using Neusilin (LSC-N1 to LSC-N8) and Fugicalin (LSC-F1 to LSC-F8). The physicochemical evaluation of all formulations exhibited well within the specification limits with respect to weight variation, hardness, friability and content uniformity. The In-vitro drug release from these LSC was evaluated in 0.1 N HCl and the optimized formulation (LSC-N8) was compared with pure drug (capsule) and physical mixture (tablet). The release studies proved that the liquisolid tablets results in higher release profile than pure drug and physical mixture due to increase in surface and wetting properties of the drug.Conclusion: LSC technique confirmed the enhanced dissolution rate of Efavirenz, which in turn helps in improving bioavailability.Â

SYNTHESIS AND CHARACTERIZATION OF POLY D-L LACTIDE (PLA) NANOPARTICLES FOR THE DELIVERY OF QUERCETIN

Objectives: Synthesis and optimization of Poly D-L Lactide (PLA) nanoparticles for the delivery of an antioxidant molecule quercetin.Methods: The quercetin was encapsulated by PLA nanoparticles by nanoprecipitation method. The average particle size and the electric charge for different formulations were measured by particle size and zeta potential analyzer. The quercetin loaded PLA nanoparticles (Q-PLAN) was characterized by differential scanning calorimetry & Fourier transform-Infra red spectroscopy, Scanning electron microscopy and Atomic force microscopy. The average drug content, encapsulation efficiency and drug release studies were carried out for different formulations of Q-PLAN. The antioxidant activity of the formulated Q-PLAN nanoparticles was tested using DPPH assay.Results: The formulation F3 (Quercetin 75 mg: PLA 200 mg) was found to be optimized formulation based on particle size analysis, Zeta potential, drug content, encapsulation efficiency and drug release studies. The mean diameter and zeta potential of optimized Q-PLAN and PLA nanoparticles were found to be 242±20 nm, 185±10 nm and-22.5±1.5 mV,-20.5±1.0 mV. The F3 formulation showed encapsulation efficiency of 73.3% and 5.5±0.06 mg/ml of actual drug loading. The F3 formulations showed 99.7% of drug release. The optimized Q-PLAN showed better scavenging effects when compared to the free quercetin.Conclusions: The poor aqueous solubility and stability of the antioxidant molecule quercetin have been improved by entrapping the quercetin molecules into the PLA nanoparticles.Â

High-Level Production of 4-Hydroxyvalerate from Levulinic Acid via Whole-Cell Biotransformation Decoupled from Cell Metabolism

gamma-Hydroxyvalerate (4HV) is an important monomer used to produce various valuable polymers and products. In this study, an engineered 3-hydroxybutyrate dehydrogenase that can convert levulinic acid (LA) into 4HV was co-expressed with a cofactor (NADH) regeneration system mediated by an NAD(+)-dependent formate dehydrogenase (CbFDH) in the Escherichia coli strain, MG1655. The resulting strain produced 23-fold more 4HV in a shake flask. The 4HV production was not dependent on ATP and required low aeration; all of these are considered beneficial characteristics for the production of target compounds, especially at an industrial scale. Under optimized conditions in a 5 L fermenter, the titer, productivity, and molar conversion efficiency for 4HV reached 100 g/L, 4.2 g/L/h, and 92%, respectively. Our system could prove to be a promising method for the large-scale production of 4HV from LA at low-cost and using a renewable biomass source