Nanosuspension of efavirenz for improved oral bioavailability: formulation optimization,in vitro,in situ and in vivo evaluation

Context: Nanosuspensions (NSs) of poorly water-soluble drugs are known to increase the oral bioavailability.

Objectives: The purpose of this study was to develop NS of efavirenz (EFV) and to investigate its potential in enhancing the oral bioavailability of EFV.

Materials and methods: EFV NS was prepared using the media milling technique. The Box–Behnken design was used for optimization of the factors affecting EFV NS. Sodium lauryl sulfate and PVP K30 were used to stabilize the NS. Freeze-dried NS was completely re-dispersed with double-distilled filtered water.

Results: Mean particle size and zeta potential of the optimized NS were found to be 320.4?±?3.62?nm and –32.8?±?0.4 mV, respectively. X-ray diffraction and differential scanning calorimetric analysis indicated no phase transitions. Rate and extent of drug dissolution in the dissolution medium for NS was significantly higher compared to marketed formulation. The parallel artificial membrane permeability assay revealed that NS successfully enhanced the permeation of EFV. Results of in situ absorption studies showed a significant difference in absorption parameters such as K_a, t_1/2 and uptake percentages between lyophilized NS and marketed formulation of EFV. Oral bioavailability of EFV in rabbits resulting from NS was increased by 2.19-fold compared to the marketed formulation.

Conclusion: Thus, it can be concluded that NS formulation of EFV can provide improved oral bioavailability due to enhanced solubility, dissolution velocity, permeability and hence absorption.     

Ethanol-mediated neutrophil extravasation in feline pancreas

Ethanol is a common cause of both acute and chronic pancreatitis. Studies in other organs suggest that polymorphonuclear neutrophils activated by ethanol may cause tissue injury in a variety of conditions. The aim of this study was to investigate the effects of ethanol on neutrophil extravasation in the feline pancreas. Pancreata were isolated and perfused at different flow rates with varying concentrations of ethanol in either a physiological or neutrophil depleted perfusate. Neutrophil extravasation was assessed by measuring pancreatic tissue myeloperoxidase (MPO) activity. Ethanol at 2.5% (54.25 mmol/liter) was the lowest concentration that still caused significant neutrophil extravasation (3.1+/-0.8 vs 1.9+/-0.2 units, P<0.05) and was accompanied by an increase in vascular resistance of 15%. Reduction of pancreatic perfusion by 15% did not significantly increase neutrophil extravasation. (1.1+/-0.3 vs 1.6+/-0.2 units, NS) Perfusion of the pancreas with neutrophil-depleted blood containing either ethanol or saline, followed by perfusion with an ethanol-free perfusate, showed an increase in neutrophil extravasation in the ethanol group compared to the control group (3.2+/-0.9 vs 1.9+/-0.2 units, P<0.05). In conclusion, ethanol causes neutrophil extravasation in the feline pancreas independent of blood flow changes and occurs despite the absence of direct neutrophil exposure to ethanol.     

Tyrp1 Mutant Variants Associated with OCA3: Computational Characterization of Protein Stability and Ligand Binding

Oculocutaneous albinism type 3 (OCA3) is an autosomal recessive disorder caused by mutations in the TYRP1 gene. Tyrosinase-related protein 1 (Tyrp1) is involved in eumelanin synthesis, catalyzing the oxidation of 5,6-dihydroxyindole-2-carboxylic acid oxidase (DHICA) to 5,6-indolequinone-2-carboxylic acid (IQCA). Here, for the first time, four OCA3-causing mutations of Tyrp1, C30R, H215Y, D308N, and R326H, were investigated computationally to understand Tyrp1 protein stability and catalytic activity. Using the Tyrp1 crystal structure (PDB:5M8L), global mutagenesis was conducted to evaluate mutant protein stability. Consistent with the foldability parameter, C30R and H215Y should exhibit greater instability, and two other mutants, D308N and R326H, are expected to keep a native conformation. SDS-PAGE and Western blot analysis of the purified recombinant proteins confirmed that the foldability parameter correctly predicted the effect of mutations critical for protein stability. Further, the mutant variant structures were built and simulated for 100 ns to generate free energy landscapes and perform docking experiments. Free energy landscapes formed by Y362, N378, and T391 indicate that the binding clefts of C30R and H215Y mutants are larger than the wild-type Tyrp1. In docking simulations, the hydrogen bond and salt bridge interactions that stabilize DHICA in the active site remain similar among Tyrp1, D308N, and R326H. However, the strengths of these interactions and stability of the docked ligand may decrease proportionally to mutation severity due to the larger and less well-defined natures of the binding clefts in mutants. Mutational perturbations in mutants that are not unfolded may result in allosteric alterations to the active site, reducing the stability of protein-ligand interactions.     

Morphology of Cereal Starches Grafted with Polyacrylonitrile

Morphological aspects of six cereal starches and their graft copolymers with polyacrylonitriles (PAN) prepared from granular as well as geletinized starches are reported. Scanning electron microscopy showed that grafting is not uniform on the surface but has also occurred in the interior of the granules. The graft copolymers prepared from gelatinized starches maintained the individuality but no resemblance with original shape of the granules.     

Time temperature equivalence in plasticized PVC

The viscoelastic response of plasticized PVC was determined from stress-relaxation data over a wide range of time at different temperatures. The relaxation modulus as a function of time was determined from these data. The relaxation curves were then shifted horizontally to obtain a master curve at a reference temperature. The amount of shift was evaluated using the WLF equation. The coefficients C and C used in the equation for the PVC were determined according to the method of reduced variables.     

Glass fiber-reinforced epoxy composites

Glass fiber-reinforced epoxy composites were prepared from the matrix resins tetraglycidyl diaminodiphenylmethane

1 Systematic name: N,N,N′,N′-Tetrakis(2,3-epoxypropyl)-4,4′-diaminodiphenylmethane.

(TGDDM) and tetraglycidyl bis(o-toluidino)-methane

2 Systematic name: N,N,N′,N′-Tetrakis(2,3-epoxypropyl)-4,4′-bis(o-toluidino)methane.

(TGMBT) using various amines like 4,4′-diaminodiphenylmethane (DDM), 4,4′-diaminodiphenylsulfone (DDS) and diethylene triamine (DETA) as curing agents. The fabricated laminates were evaluated for their mechanical and dielectrical properties and chemical resistance. The composites prepared using an epoxy fortifier (20 phr) showed significant improvement in the mechanical properties.     

Acrylic Homo- and Co-polymers Based on 2, 4-dichlorophenyl Methacrylate and 8-quinolinyl Methacrylate

The monomer 2, 4-dichlorophenyl methacrylate (2,4-DMA) was synthesized from 2, 4-dichlorophenol and characterized by conventional methods. The homopolymers of 2,4-dichlorophenyl methacrylate and its copolymers with 8-quinolinyl methacrylate (8-QMA) in different feed ratio were prepared by free radical polymerization using dimethyl formamide (DMF) as a solvent and 2,2-azobis iso butyronitrile (AIBN) as an initiator. The resulting polymers were characterized by using IR spectroscopy. Reactivity ratios of monomer were evaluated using UV-spectroscopy. Average molecular weights and polydispersity were determined by gel permeation chromatography (GPC). Solution viscosity was also obtained. The thermal analysis was carried out using TGA and DSC. The homo- and co-polymers were also tested for their antimicrobial properties against selected microorganism. The metal ion uptake capacity of synthesized copolymers was estimated by batch equilibration method using different metal ion solution under different experimental conditions. It is observed that due to the presence of pendant ester bound quinoline group, the copolymers are capable of adsorbing cations from their aqueous solution.     

Morphological, mechanical and gas-transport characteristics of crosslinked poly(propylene glycol): homopolymers, nanocomposites and blends

Linear polyethers possess unusually high CO₂ solubility and, hence, selectivity due to the presence of accessible ether linkages that can interact with the quadrupolar moment of CO₂ molecules. In this work, membranes derived from crosslinked poly(propylene glycol) diacrylate (PPGda) oligomers differing in molecular weight (M), as well as PPGda nanocomposites containing either an organically-modified montmorillonite clay or a methacrylate-terminated fumed silica are investigated and compared with highly CO₂-selective poly(ethylene glycol) diacrylate (PEGda) homopolymer and nanocomposite membranes previously reported. The rheological and permeation properties of PPGda depend sensitively on M, with the elastic modulus decreasing, but CO₂ permeability and CO₂/H₂ selectivity increasing, with increasing M. Incorporation of either nanofiller into PPGda enhances the elastic modulus and reduces the gas permeability in the resultant nanocomposites without strongly affecting CO₂/H₂ selectivity. Blending PPGda and PEGda prior to chemical crosslinking yields binary membranes that exhibit intermediate gas-transport properties accurately described by a linear rule of mixtures.     

Reactive melt modification of polypropylene with a crosslinkable polyester

Melt blending of polypropylene (PP) with a low molecular weight (MW) crosslinkable unsaturated polyester (UP) was studied in a batch mixer and a twin‐screw extruder in the presence of peroxide (POX) free radical initiator. Competing degradation and crosslinking reactions of the peroxide with the blend components leading to significant change in their viscosity ratio were studied and controlled during the blending process. Rheological studies show that depending on the process conditions some reacted PP/UP blends have a pronounced suspension behavior due to the presence of the dispersed polyester gel particles in a low MW PP matrix. It is also evident from SEM analysis that the reacted blends have a more uniform and finer morphology than the unreacted ones. The blends are also characterized by FTIR, which strongly suggests the presence of “block” or “graft” PP‐UP structures that may enhance phase interaction and promote compatibility in the reacted PP/UP blends. Such blends are considered as suitable compatibilizers of PP/high MW thermoplastic polyester blends and as modifiers for low density extrusion foaming of similar blends.