Palladium oxide-decorated mesoporous silica on graphene oxide nanosheets as a heterogeneous catalyst for the synthesis of ß-substituted indole derivatives.

In this work, an efficient and facile strategy has been adopted for the stepwise synthesis of the RGO-MSiO2/PdO hybrid nanomaterial (HY-NM). Herein, a hybrid nanostructure of mesoporous silica over graphene oxide (GO) sheets has been developed followed by immobilizing palladium oxide nanoparticles (PdO NPs), and then it has been utilized for catalyzing a multicomponent reaction (MCR). To authenticate the successful synthesis of the HY-NM and successive immobilization of PdO NPs, various physicochemical characterization techniques were utilized such as SEM, EDAX, HR-TEM, HR-XRD, TGA, BET, FT-IR, and XPS analysis. The activity of the HY-NM has been determined by performing the catalyst-mediated synthesis of ß-substituted indole derivatives (yield 90-98%). The excellent catalytic activity of the prepared HY-NM could be observed due to its high surface area and large porosity, which facilitates the penetration and interaction of reactant molecules with the catalytic active species. This protocol eliminates the requirement of further purification after the isolation of the product from the reaction mixture. The ease of handling, recyclability of the catalyst, and simple work-up procedure are the main features of this protocol. The synthesized HY-NM could be recycled for multiple catalytic cycles making it a very effective heterogeneous catalyst.

Ru x Pd y Alloy Nanoparticles Uniformly Anchored on Reduced Graphene Oxide Nanosheets (Ru x Pd y @rGO): A Recyclable Catalyst.

In this work, Ru x Pd y alloy nanoparticles were uniformly decorated on a two-dimensional reduced graphene oxide (rGO) sheet by an in situ chemical co-reduction process. The resulting products were characterized by various physiochemical techniques such as X-ray diffraction, Raman spectroscopy, energy-dispersive X-ray spectroscopy, inductively coupled plasma atomic absorption spectroscopy, X-ray photoelectron spectroscopy, and transmission electron microscopy. Further, the synthesized Ru x Pd y @rGO nanocomposites have been employed as a heterogeneous catalyst for three different catalytic reactions: (1) dehydrogenation of aqueous ammonia borane (AB); (2) hydrogenation of aromatic nitro compounds using ammonia borane as the hydrogen source, and (3) for the synthesis of aromatic azo derivatives. The present work illustrates the sustainable anchoring of metal nanoparticles over the surface of rGO nanosheets, which could be used for multifarious catalytic reactions.

Influence of pH, ß-Cyclodextrin, and Metal Ions on the Solubility and Stability of the Medicinally Competent Isoxazole Derivative of Curcumin: A Photophysical Study.

The ß-diketo-modified isoxazole derivative of curcumin (IOC) is well renowned for its anticancer, antioxidant, antimalarial, antiproliferative, and many other biological activities. With the aim of obtaining fundamental knowledge on the photophysics of IOC, the present work was directed toward delineating those at different pH environments and studying the degradation profiles of IOC at five different pH values. Because one of the primary drawbacks of curcumin is its rapid degradation at physiological conditions, the studies showed that the problem could be resolved, as the IOC molecule was extremely stable even in a highly alkaline medium. Further, in order to encounter the problems associated with the low solubility of IOC in aqueous media, ß-CD (ß-cyclodextrin) was used and calculations of the thermodynamic parameters revealed that the process of development of the host-guest inclusion complex was highly spontaneous in nature. The synthesis of the IOC:ß-CD inclusion complex has also been accomplished in the solid state, and the solid formed has been characterized using various physicochemical techniques. Finally, while variations in the pH as well as addition of foreign metal ions in +1 and +2 oxidation states showed minimal effect on the photophysics of the IOC:ß-CD inclusion complex, antiproliferative studies performed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays revealed their nontoxic nature on fibroblast L929 normal cell lines and extremely toxic activity on human lung cancer A549 cell lines.

The Role of Imidazolium-Based Surface-Active Ionic Liquid to Restrain the Excited-State Intramolecular H-Atom Transfer Dynamics of Medicinal Pigment Curcumin: A Theoretical and Experimental Approach.

The naturally occurring polyphenolic compound curcumin has shown various medicinal and therapeutic effects. However, there are various challenges associated with curcumin, which limits its biomedical applications, such as its high degradation rate and low aqueous solubility at neutral and alkaline pH. In the present study, efforts have been directed towards trying to resolve such issues by encapsulating curcumin inside the micelles formed by imidazolium-based surface-active ionic liquid (SAIL). The shape and size of the micelles formed by the SAIL have been characterized by using DLS analysis as well as TEM measurements. The photo-physics of curcumin in the presence of ionic liquid (IL) and also with the addition of salt (NaCl) has been explored by using different optical spectroscopic tools. The time-dependent absorption studies have shown that there is relatively higher suppression in the degradation rate of curcumin after encapsulation by the imidazolium-based SAIL in an aqueous medium. The TCSPC studies have revealed that there is deactivation in the nonradiative intramolecular hydrogen transfer process of curcumin in the presence of IL micelles as well as with the addition of salt. Furthermore, the time-dependent fluorescence anisotropy measurement has been carried out to figure out the location of curcumin inside the micellar system. In order to correlate all experimental findings, density functional theory (DFT) and classical molecular dynamics (MD) simulations at neutral pH media have been performed. It has been found that the van der Waals force of interactions plays a major role in the stabilization of curcumin in the micelles rather than the coulombic forces. It also has been observed that the van der Waals interactions remain unaffected in the presence of salt. However, as revealed by the MD simulation results, the micelles are found to be more compact in size after the addition of salt. The RMSD results show that the micelles formed by the SAIL achieve greater stability after a particular time constraint. Our results have divulged that the SAIL could act as a promising drug delivery system.

Synthesis of dendritic fibrous nanosilica over a cubic core (cSiO2@DFNS) with catalytically efficient silver nanoparticles for reduction of nitroarenes and degradation of organic dyes.

In this study, dendritic fibrous core-shell silica particles having cubic morphology with uniform and vertical nanochannels have been successfully synthesised. The synthesized dendritic fibrous nanosilica over a cubic core (cSiO2@DFNS) have been characterized by using various techniques, such as powder X-ray diffraction, TEM, FE-SEM, TGA EDS, FT-IR and N2 adsorption-desorption experiments. The prepared DFNS particles demonstrated a very high surface area and pore diameter. Amine groups were functionalized on the fibres of cSiO2@DFNS and after that silver nanoparticles could be successfully immobilized on amine functionalized cubic silica particles. Due to the presence of a high surface area and a uniform pore diameter, the silver nanoparticle loaded cSiO2@DFNS could be successfully employed as an efficient and recoverable catalyst for reduction of toxic aromatic nitro compounds and degradation of organic dyes. Higher catalytic activity of the prepared material could be attributed to its fibrous morphology which could facilitate proper interactions of the reactants molecules with the silver nanoparticles.

pH-dependent immobilization of urease on glutathione-capped gold nanoparticles.

Urease is a nickel-dependent metalloenzyme that catalyzes the hydrolysis of urea to form ammonia and carbon dioxide. Although the enzyme serves a significant role in several detoxification and analytical processes, its usability is restricted due to high cost, availability in small amounts, instability, and a limited possibility of economic recovery from a reaction mixture. Hence, there is a need to develop an efficient, simple, and reliable immobilization strategy for the enzyme. In this study, the carboxyl terminated surface of glutathione-capped gold nanoparticles have been utilized as a solid support for the covalent attachment of urease. The immobilization has been carried out at different pH conditions so as to elucidate its effect on the immobilization efficiency and enzyme bioactivity. The binding of the enzyme has been quantitatively and qualitatively analyzed through techniques like ultraviolet-visible spectroscopy, intrinsic steady state fluorescence, and circular dichorism. The bioactivity of the immobilized enzyme was investigated with respect to the native enzyme under different thermal conditions. Recyclability and shelf life studies of the immobilized enzyme have also been carried out. Results reveal that the immobilization is most effective at pH of 7.4 followed by that in an acidic medium and is least in alkaline environment. The immobilized enzyme also exhibits enhance activity in comparison to the native form at physiological temperature. The immobilized urease (on gold glutathione nanoconjugates surface) can be effectively employed for biosensor fabrication, immunoassays and as an in vivo diagnostic tool in the future.

Synthesis of thermoresponsive polymers for drug delivery.

A protocol for synthesizing thermosensitive copolymers of N-isopropylacrylamide (NIPAM) and N-vinylpyrrolidone (VP), cross-linked with N,N'-methylene-bis-acrylamide (MBA) has been described in this chapter. The copolymers have been formed at different concentrations of NIPAM and VP and at two different temperatures (70 °C and 30 °C). The lower critical solution temperature (LCST) of the samples has been measured, and the size of the particles formed with the highest concentration of NIPAM and lowest concentration of VP (MG1 and NG1) has been measured at three different temperatures of 25 °C, 35 °C, and 37 °C. Both MG1 and NG1 showed the lowest size at 37 °C. The MG1 and NG1 samples were further characterized using TEM and SEM. The MG1 particles were subsequently used for protein drug delivery, using BSA as a model. The release profile showed the best fit with the zero-order model. Finally, cytotoxicity studies of the synthesized MG1 and NG1 particles were carried out, using in vitro MTT assay, so as to determine the overall biocompatibility of the materials.

Synthesis of a smart nanovehicle for targeting liver.

A protocol for the synthesis of a smart drug delivery system based on gold nanoparticles has been described in this chapter. The synthesized drug delivery system has been shown to release the bioactive material in response to an intracellular stimulus (glutathione concentration gradient) and hence shown to behave in an intelligent manner. Gold nanoparticles have been employed as the core material with the surface functionalities of thiolated PEG. PEG owing to its non-immunogenicity and non-antigenicity would impart considerable stability and longer in vivo circulation time to the gold nanoparticles. The end groups of PEG chains have been derivatized with functional groups like aldehyde (-CHO) and amine (-NH2) which could behave as flexible arms for the attachment of "target specific ligands" and other bioactive substances. Lactose, a liver targeting ligand, has been employed as the target specific moiety. A Coumarin derivative has been synthesized and used as the model fluorescent tag as well as a linker to examine the glutathione-mediated release through fluorescence spectroscopy and for the conjugation of bioactive molecules, respectively. A check for the cytocompatibility of the synthesized nanovehicle on the cultured mammalian cell lines has also been carried out. Finally, in the latter parts of the chapter (mimicking the in vivo conditions), time-dependent in vitro release of the model fluorescent moiety has also been analyzed at different glutathione concentrations.

Synthesis of a smart gold nano-vehicle for liver specific drug delivery.

Targeting drug formulations to specific tissues and releasing the bioactive content in response to a certain stimuli remains a significant challenge in the field of biomedical science. We have developed a nanovehicle that can be used to deliver "drugs" to "specific" tissues. For this, we have simultaneously modified the surface of the nanovehicle with "drugs" and "tissue-specific ligands". The "tissue-specific ligands" will target the nanovehicle to the correct tissue and release the "drug" of interest in response to specific stimuli. We have synthesised a "lactose surface-modified gold nanovehicle" to target liver cells and release the model fluorescent drug (coumarin derivative) in response to the differential glutathione concentration (between blood plasma and liver cells). Lactose is used as the liver-specific targeting ligand given the abundance of L-galactose receptors in hepatic cells. The coumarin derivative is used as a fluorescent tag as well as a linker for the attachment of various biologically relevant molecules. The model delivery system is compatible with a host of different ligands and hence could be used to target other tissues as well in future. The synthesised nanovehicle was found to be non-toxic to cultured human cell lines even at elevated non-physiological concentrations as high as 100 µg/mL. We discover that the synthesised gold-based nanovehicle shows considerable stability at low extracellular glutathione concentrations; however coumarin is selectively released at high hepatic glutathione concentration.

Synthesis and characterization of thermoresponsive copolymers for drug delivery.

We report the synthesis and characterization of two nontoxic, thermogelling drug delivery systems which are liquid at room temperatures but become a gel at physiological temperature (37°C) potentially leading to release of a drug molecule. We selected temperature as the stimulus for drug release as it is physiologically invariant. A free radical polymerization of N-isopropylacrylamide (NIPAM) and N-vinylpyrrolidone (VP) was carried out under nitrogen atmosphere in double-distilled water at two different temperatures (30°C and 70°C), and the copolymers obtained were characterized by various analytical techniques. The molar ratios of the two monomers were altered with increasing NIPAM content and their cloud point temperature or least critical solution temperature (LCST) was determined. The copolymer at 9:1 ratio of NIPAM to VP resulted in the formation of nanoparticle-based gel (NG1) at 30°C; however, at 70°C, a microgel (MG1) was formed. The LCST of the nanogel and microgel was 33.5-34°C and 36.5-37°C, respectively. Thus, both the copolymers are water soluble at room temperature, but distinct phases appear at physiological temperatures. We hypothesized that these copolymers on entrapment with a drug could be used for topical application to the skin or eye for controlled drug delivery applications. Toxicological studies revealed that the copolymers are nontoxic in HeLa cells. Finally, our experiments show that a model drug [bovine serum albumin (BSA)] is released at 37°C with zero-order kinetics and confirmed using multiple well-known mathematical models.

Biginelli reaction catalyzed by copper nanoparticles.

We recently reported a novel synthesis of copper nanoparticles from copper sulphate utilizing the charge-compensatory effect of ionic liquid [bmim]BF(4) and ethylene glycol. The nanoparticles were characterized and found to be stable for one year. Here we hypothesize that the stabilized nanoparticles should be able to catalyze one-pot multicomponent organic reactions. We show that the nanoparticles catalyzed Biginelli reaction at room temperature to give the product 3,4-dihydopyrimidinone (>90% yield in ~15 minutes) from aldehydes, ß-diketoester (ethylacetoacetate) and urea (or thiourea). ). Remarkably, such high yields and rapid kinetics was found to be independent of the electronic density on the reactant aryl-aldehyde. This was probably because even the surface-active particles reacted faster in the presence of ionic liquid as compared to conventional methods. The heterocyclic dihydropyrimidinones (DHPMs) and their derivatives are widely used in natural and synthetic organic chemistry due to their wide spectrum of biological and therapeutic properties (resulting from their antibacterial, antiviral, antitumor and anti-inflammatory activities. Our method has an easy work-up procedure and the nanoparticles could be recycled with minimal loss of efficiency.

Using hydrophilic ionic liquid, [bmim]BF4-ethylene glycol system as a novel media for the rapid synthesis of copper nanoparticles.

In this work, we present a novel method for the synthesis of copper nanoparticles. We utilize the charge compensatory effect of ionic liquid [bmim]BF(4) in conjunction with ethylene glycol for providing electro-steric stabilization to copper nanoparticles prepared from copper sulphate using hydrazine hydrate as a reducing agent. The formed copper nanoparticles showed extended stability over a period of one year. Copper nanoparticles thus prepared were characterized by powder X-ray diffraction measurements (pXRD), transmission electron microscopy (TEM) and quasi elastic light scattering (QELS) techniques. Powder X-ray diffraction (pXRD) analysis revealed relevant Bragg's reflection for crystal structure of copper. Powder X-ray diffraction plots also revealed no oxidized material of copper nanoparticles. TEM showed nearly uniform distribution of the particles in methanol and confirmed by QELS. Typical applications of copper nanoparticles include uses in conductive films, lubrication and nanofluids. Currently efforts are under way in our laboratory for using these nanoparticles as catalysts for a variety of organic reactions.

A rapid and simple route for the synthesis of lead and palladium nanoparticles in tetrazolium based ionic liquid.

In the present work, we report a novel method for the synthesis of palladium and lead nanoparticles by the reduction method in tetrazolium ring based ionic liquid. Palladium and lead nanoparticles so-prepared were well characterized by powder X-ray diffraction measurements (pXRD), transmission electron microscopy (TEM) and quasi elastic light scattering (QELS) techniques. Powder X-ray diffraction (pXRD) analysis revealed all relevant Bragg's reflection for crystal structure of palladium and lead. Powder X-ray diffraction plots also revealed no oxidized material of palladium and lead nanoparticles. TEM showed nearly uniform distribution of the particles in methanol and confirmed by QELS. Typical applications of palladium nanoparticles include in vitro use and sensor design applications. Palladium nanoparticles is also ideal for spin coating, self-assembly and monolayer formation. Palladium nanoparticles can also be considered as potential new catalysts.

Biodegradable calcium phosphate nanoparticle with lipid coating for systemic siRNA delivery.

A lipid coated calcium phosphate (LCP) nanoparticle (NP) formulation was developed for efficient delivery of small interfering RNA (siRNA) to a xenograft tumor model by intravenous administration. Based on the previous formulation, liposome-polycation-DNA (LPD), which was a DNA-protamine complex wrapped by cationic liposome followed by post-insertion of PEG, LCP was similar to LPD NP except that the core was replaced by a biodegradable nano-sized calcium phosphate precipitate prepared by using water-in-oil micro-emulsions in which siRNA was entrapped. We hypothesized that after entering the cells, LCP would de-assemble at low pH in the endosome, which would cause endosome swelling and bursting to release the entrapped siRNA. Such a mechanism was demonstrated by the increase of intracellular Ca(2+) concentration as shown by using a calcium specific dye Fura-2. The LCP NP was further modified by post-insertion of polyethylene glycol (PEG) with or without anisamide, a sigma-1 receptor ligand for systemic administration. Luciferase siRNA was used to evaluate the gene silencing effect in H-460 cells which were stably transduced with a luciferase gene. The anisamide modified LCP NP silenced about 70% and 50% of luciferase activity for the tumor cells in culture and those grown in a xenograft model, respectively. The untargeted NP showed a very low silencing effect. The new formulation improved the in vitro silencing effect 3-4 folds compared to the previous LPD formulation, but had a negligible immunotoxicity.

Lipid-based systemic delivery of siRNA.

RNAi technology has brought a new category of treatments for various diseases including genetic diseases, viral diseases, and cancer. Despite the great versatility of RNAi that can down regulate almost any protein in the cells, the delicate and precise machinery used for silencing is the same. The major challenge indeed for RNAi-based therapy is the delivery system. In this review, we start with the uniqueness and mechanism of RNAi machinery and the utility of RNAi in therapeutics. Then we discuss the challenges in systemic siRNA delivery by dividing them into two categories-kinetic and physical barriers. At the end, we discuss different strategies to overcome these barriers, especially focusing on the step of endosome escape. Toxicity issues and current successful examples for lipid-based delivery are also included in the review.

Preparation, characterization and in vitro drug release studies of novel polymeric nanoparticles.

Polymeric nanoparticles of AADG cross-linked with MBA encapsulating water soluble macromolecules such as FITC-Dextran have been prepared in the reverse micellar system. The particles obtained were of >85nm in diameter which were highly monodisperse. An optically clear solution was obtained on redispersing these nanoparticles in aqueous buffer. Size and morphology of the particles remains the same on re-dispersing the lyophilized powder of these nanoparticles in aqueous buffer. The size dependency of the particles on the monomer and surfactant concentration was observed. The average size of the nanoparticles as obtained from DLS studies ranges from 74 to 114nm in case 0.06M AOT and 62-104nm in case of 0.1M AOT concentration. FITC-Dextran was entrapped into nanoparticles with high efficiency (>70%). The pH dependent release of the entrapped molecules from these nanoparticles was also studied. At pH 5.0 solution, approximately 43% of FITC-Dx was released and at pH 7.4 it was about 70%.

Ultra-low sized cross-linked polyvinylpyrrolidone nanoparticles as non-viral vectors for in vivo gene delivery.

In principle, the technique of gene delivery involves taking complete or parts of genes that can code specific message and delivering them to selected cells in the body. Such a transfer of plasmid DNA into mammalian cells has posed major challenges for gene therapy. This study shows the encapsulation of a plasmid DNA in cross-linked polyvinylpyrrolidone (PVP) nanoparticles of size less than 100 nm. This kind of encapsulation provides complete protection to the plasmid DNA from external DNase environment and generates the hope that the resulting formulation can be developed into a potential vector for effective gene delivery. In order to check this potentially, the reporter gene pSVbeta-gal was encapsulated and in vitro transfection efficiency of this system was found to be nearly 80% compared to the commercially available transfection reagent Polyfect. Further, in vivo biodistribution studies indicated that this system could be used safely for effective gene delivery.

Cross-linked polyvinylpyrrolidone nanoparticles: a potential carrier for hydrophilic drugs.

Injectable hydrogel polymeric nanoparticles of polyvinylpyrrolidone cross-linked with N,N'-methylene bis-acrylamide and encapsulating water-soluble macromolecules such as FITC-dextran (FITC-Dx) have been prepared in the aqueous cores of reverse micellar droplets. These particles are 100 nm and below in diameter with a narrow size distribution. When dispersed in aqueous buffer these particles appear to be transparent and give an optically clear solution. Lyophilized powder of these nanoparticles is redispersable in aqueous buffer without any change in the size and morphology of the particles. The efficiency of FITC-Dx entrapment by these nanoparticles is high (>70%) and depends on the amount of cross-linking agent present in the polymeric material. The release of the entrapped molecules from these nanoparticles depends on the degree of cross-linking of the polymer, particle size, pH of the medium, and extent of loading, as well as temperature.

Calcium phosphate nanoparticles as novel non-viral vectors for targeted gene delivery.

Calcium phosphate nanoparticles present a unique class of non-viral vectors, which can serve as efficient and alternative DNA carriers for targeted delivery of genes. In this study we report the design and synthesis of ultra-low size, highly monodispersed DNA doped calcium phosphate nanoparticles of size around 80 nm in diameter. The DNA encapsulated inside the nanoparticle is protected from the external DNase environment and could be used safely to transfer the encapsulated DNA under in vitro and in vivo conditions. Moreover, the surface of these nanoparticles could be suitably modified by adsorbing a highly adhesive polymer like polyacrylic acid followed by conjugating the carboxylic groups of the polymer with a ligand such as p-amino-1-thio-beta-galactopyranoside using 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride as a coupling agent. We have demonstrated in our studies that these surface modified calcium phosphate nanoparticles can be used in vivo to target genes specifically to the liver.