[show abstract][hide abstract] ABSTRACT: A polarization study carried out on a thin supported liquid membrane separating two aqueous compartments is presented. Transfer of both the ionized and uncharged form of an organic tracer dye, rhodamine B ([9-(2-carboxyphenyl)-6-diethylamino-3-xanthenylidene]-diethylammonium chloride), across supported liquid membranes composed of one of 1-octanol (octan-1-ol), 1,9-decadiene (deca-1,9-diene), 1,2-dichlorobenzene or nitrophenyl octyl ether (1-(2-nitrophenoxy)octane), was studied using cyclic voltammetry and UV-vis absorption spectrophotometry. Concentration analysis indicates that the high membrane concentration of rhodamine B determines the ionic transfer observed via voltammetry, which is consistent with the low aqueous ionic concentration and large membrane/aqueous distribution of the molecule. The observed double-transfer voltammogram, although it has been largely neglected in previous literature, is a logical consequence of the presence of two liquid-liquid interfaces and is rationalised in terms of ion transfer across the two interfaces on either side of the membrane and supported by voltammograms obtained for a series of ions of varied lipophilicity. The bipolar nature of the voltammetric response offers an effective way of mass transport control via changing polarity of the applied voltage and finds immediate use in extraction, purification and separation applications.
[show abstract][hide abstract] ABSTRACT: ATR is an attractive new anticancer drug target whose inhibitors have potential as chemo- or radiation sensitizers or as monotherapy in tumors addicted to particular DNA-repair pathways. We describe the discovery and synthesis of a series of sulfonyl-morpholino-pyrimidines which show potent and selective ATR inhibition. Optimization from a high quality screening hit within tight SAR space led to compound 6 (AZ20) which inhibits ATR immunoprecipitated from HeLa nuclear extracts with an IC50 of 5 nM and ATR mediated phosphorylation of Chk1 in HT29 colorectal adenocarcinoma tumor cells with an IC50 of 50 nM. Compound 6 potently inhibits the growth of LoVo colorectal adenocarcinoma tumor cells in-vitro and has high free exposure in mouse following moderate oral doses. At well tolerated doses 6 leads to significant growth inhibition of LoVo xenografts grown in nude mice. Compound 6 is a useful compound to explore ATR pharmacology in-vivo.
Journal of Medicinal Chemistry 02/2013; · 5.61 Impact Factor
[show abstract][hide abstract] ABSTRACT: The purpose of this study is to develop a droplet-based microfluidic device capable of monitoring drug precipitation upon a shift from gastric pH (pH 1.5) to intestinal pH (pH 6.5-7.0). The extent of precipitation occurring in droplets over time was measured using a novel on-chip laser scattering technique specifically developed for this study. The precipitation of ketoconazole, a poorly water-soluble basic drug, was investigated under different concentrations and pH values. It has been shown that the drug precipitates rapidly under supersaturation. Two water-soluble aqueous polymers, namely, polyvinylpyrrolidone (PVP) and hydroxypropylmethylcellulose (HPMC) have been evaluated as precipitation inhibitors. HPMC was shown to be the most potent precipitation inhibitor. It is envisaged that the microfluidic pH-shift method developed in this study would form a proof-of-concept study, towards the development of a high throughput method for screening pharmaceutical excipients/precipitation inhibitors.
The Analyst 01/2013; 138:339-345. · 4.23 Impact Factor
[show abstract][hide abstract] ABSTRACT: The stepwise hydrogenation of 9-ethylcarbazole to 9-ethyl-perhydrocarbazole (Pl 12[H]) via partially hydrogenated intermediate(s) was studied over a number of supported ruthenium and rhodium catalysts. The reaction pathways were modelled and the rate constants for individual hydrogenation steps were compared. It was found that the selectivities to the reaction intermediates and products were highly dependent on the electronic structure of the particular metal and the nature of the support used. Ruthenium was found to be the most active metal for this reaction but it suffered from a poor selectivity to the desired product due to the formation of a kinetically stable intermediate, 9-ethyl-octahydrocarbazole (Pl 8[H]) in short reaction time. On the other hand, rhodium catalysts with moderate activity gave a higher selectivity to the fully hydrogenated product under comparable conditions. It was also found that the presence of a hydrophilic support such as alumina or rutile can give kinetically favoured cis-isomers of the 9-ethyl-perhydrocarbazole. Regarding application of the reversible hydrogen storage concept, the storage material should be able to switch between fully hydrogen loaded and unloaded forms during hydrogenation (material regeneration) and dehydrogenation (delivery of hydrogen gas) in short times. Formation of any stable intermediates and stereo-non-favoured isomers with a particular type of catalyst can result in significant implications to the overall storage capacity as well as operation times for hydrogen gas delivery and regeneration. Thus, this study yields valuable information on the suitability of various metal catalysts for use in hydrogen storage systems based on the liquid organic hydride (LOH) concept.
Energy & Environmental Science 08/2012; 5(9):8621-8630. · 11.65 Impact Factor
[show abstract][hide abstract] ABSTRACT: It is demonstrated that engineered arginase with a single protruded site of cysteine deliberately placed away from its active centre by site-directed mutagenesis can facilitate its attachment on a gold-nanoparticle surface with atomic precision, resulting in no apparent loss in enzymatic activity.
Chemical Communications 07/2012; 48(62):7693-5. · 6.38 Impact Factor
[show abstract][hide abstract] ABSTRACT: The purposes of this study are to evaluate if the PAMPA (Parallel Artificial Membrane Permeability Assay) permeability and the true partition coefficient could be useful for predicting AUC and MIC data of a group of antibacterial fluoroquinolones (FQs). The protonation macro- and microconstants, the n-octanol/water partition coefficients at isoelectric pHs, and the PAMPA permeability of 11 selected FQs were determined, and used to calculate the true partition coefficient, the interactivity parameter between the acidic and basic group, and the apparent intrinsic permeability. It has been shown that the apparent intrinsic permeability correlates well with the AUC in human, whereas the true partition coefficient and the interactivity parameter correlate with 1/MIC values on two Gram-positive bacteria, namely Streptococcus pneumonia and Staphylococcus aureus (methicillin-susceptible). The AUC/MIC ratios predicted from these correlations have shown to be in good agreement with the literature values. It is envisaged that the models described in this study could be useful in the development of new FQs by enabling an early prediction of AUC/MIC ratios based on physicochemical properties.
European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2012; 47(1):21-7. · 2.61 Impact Factor
[show abstract][hide abstract] ABSTRACT: The preparation and application of a simple silver/silver sulfate reference electrode for an aqueous solution, which can be used as an alternative in chloride-free systems, is reported. The electrodes are prepared by galvanostatic oxidation of silver wire in sulfate solution: the potential stability with time is measured as a function of the current density and overall charge used in oxidation. The electrode potential is also measured in a wide concentration range of sulfate and chloride solutions and an explanation of the observed stability is presented. The range of optimal conditions, crucial for the correct electrode operation, is discussed.
[show abstract][hide abstract] ABSTRACT: An analytical technique for the detection of permeation of a fully ionized analyte across a lipophilic membrane is reported. The system, which is comprised of two aqueous compartments (donor and acceptor) separated by a supported liquid membrane, is based on the parallel artificial membrane permeation assay (PAMPA), widely used in the drug discovery process to estimate permeability in vivo. The in situ spectroelectrochemical method developed here employs mechanical stirring of the solution phases on either side of the membrane, external polarization of the membrane, and in situ detection of the analyte via UV-vis spectrophotometry. The flux of the crystal violet cation across the membrane is simultaneously measured via UV-vis spectrophotometry and voltammetry/chronoamperometry as a function of applied potential. The relative contribution of two permeation modes, i.e., that due to naked ions and ion-pairs, is thereby quantified. The open circuit potential difference between the two aqueous compartments and the cyclic voltammetric response are also recorded as a function of time and compared with the predicted values.
[show abstract][hide abstract] ABSTRACT: Chemoembolization has been used in the field of interventional oncology. Although practiced widely, it has only recently been demonstrated that the use of transarterial chemoembolization (TACE) provides a survival benefit based on randomized controlled trials. TACE combines the effect of targeted chemotherapy with the effect of ischemic necrosis induced by arterial embolization. Most of the TACE procedures have been based on iodized oil utilizing its microembolic and drug-carrying characteristics. Recently, there have been efforts to improve the delivery of chemotherapeutic agents to a tumor, which leads to the development of drug-eluting particles. In this review, we will describe the properties and efficacy of some chemoembolization agents which are commercially available and/or currently under clinical investigations. The potential and future of this new form of transcatheter arterial therapy for liver cancer will be discussed.
European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 09/2011; 44(1-2):1-10. · 2.61 Impact Factor
[show abstract][hide abstract] ABSTRACT: The purpose of this study was to develop an in vitro permeation model that will predict the fraction of drugs absorbed in humans. A rotating-diffusion cell with two aqueous compartments, separated by a lipid-impregnated artificial membrane, was used to determine the permeability of drugs under conditions of controlled hydrodynamics. The measured effective permeability coefficient was modified to include the paracellular transport derived from a previously reported colorectal adenocarcinoma epithelial cell line (Caco-2) permeability study and the effects of unstirred water layer anticipated in vivo. Permeability data were collected for 31 different marketed drugs with known absolute oral bioavailability and human hepatic clearance data. Literature bioavailability values were corrected for the first pass hepatic clearance thus obtaining the fraction absorbed from intestinal lumen (fraction absorbed), F(a), while assuming that the fraction escaping intestinal extraction, F(g), was approximately ~1. Permeability obtained under conditions of controlled hydrodynamics was compared with the permeability measured under unstirred conditions. It is shown that the optimized effective permeability correlates with the fraction absorbed. In contrast, permeability data obtained under unstirred conditions does not show a good correlation. The in vitro permeation model developed in this study predicts the fraction absorbed of the selected drugs in humans within experimental uncertainty. It has been demonstrated that the correlation with the fraction absorbed is greatly improved using the permeability data obtained under controlled hydrodynamics with paracellular transport included in the model.
European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 08/2011; 44(3):299-309. · 2.61 Impact Factor
[show abstract][hide abstract] ABSTRACT: We show that highly porous silica-based nanoparticles prepared via micro-emulsion and sol-gel techniques are stable colloids in aqueous solution. By incorporating a magnetic core into the porous silica nano-composite, it is found that the material can be rapidly separated (precipitated) upon exposure to an external magnetic field. Alternatively, the porous silica nanoparticles without magnetic cores can be equally separated from solution by applying a high-speed centrifugation. Using these silica-based nanostructures a new high-throughput method for the determination of partition coefficient for water/n-octanol is hereby described. First, a tiny quantity of n-octanol phase is pre-absorbed in the porous silica nano-composite colloids, which allows an establishment of interface at nano-scale between the adsorbed n-octanol with the bulk aqueous phase. Organic compounds added to the mixture can therefore undergo a rapid partition between the two phases. The concentration of drug compound in the supernatant in a small vial can be determined by UV-visible absorption spectroscopy. With the adaptation of a robotic liquid handler, a high-throughput technology for the determination of partition coefficients of drug candidates can be employed for drug screening in the industry based on these nano-separation skills. The experimental results clearly suggest that this new method can provide partition coefficient values of potential drug candidates comparable to the conventional shake-flask method but requires much shorter analytical time and lesser quantity of chemicals.
Journal of Separation Science 07/2011; 34(18):2505-12. · 2.59 Impact Factor
[show abstract][hide abstract] ABSTRACT: To investigate the permeation of two ionisable drug molecules, warfarin and verapamil, across artificial membranes. For the first time since the introduction of the parallel artificial membrane permeation assay (PAMPA) in 1998, in situ permeation-time profiles of drug molecules are studied.
The method employs a rotating-diffusion cell where the donor and acceptor compartments are separated by a lipid-impregnated artificial membrane. The permeation of the solute is investigated under well-defined hydrodynamic conditions with control over the unstirred water layer. The flux of the permeating molecule is analysed in situ using UV spectrophotometry.
In situ permeation-time profiles are obtained under hydrodynamic control and used to determine permeability coefficients. An advanced analytical transport model is derived to account for the membrane retention, two-way flux and pH gradient between the two compartments. Moreover, a numerical permeation model was developed to rationalise the time-dependent permeation profiles. The membrane permeability, intrinsic permeability and unstirred water permeability coefficients of two drug molecules are obtained from two independent methods, hydrodynamic extrapolation and pH profiling, and the results are compared.
Both warfarin and verapamil exhibit high permeability values, which is consistent with the high fraction absorbed in human. Our results demonstrate that a considerable lag-time, varying with the solute lipophilicity and stirring rate, exists in membrane permeation and leads to incorrect compound ranking if it is not treated properly. Comparison of the permeability data as a function of pH and stirring rate suggests that some transport of the ionized molecules occurs, most likely via ion-pairing.
Pharmaceutical Research 05/2010; 27(8):1644-58. · 4.74 Impact Factor
[show abstract][hide abstract] ABSTRACT: The study aimed to predict effective human jejunal permeability (P(eff)) using a biophysical model based on parametrized paracellular, aqueous boundary layer, and transcellular permeabilities, and the villus-fold surface area expansion factor (k(VF)). Published human jejunal data (119 P(eff), 53 compounds) were analyzed by a regression procedure incorporating a dual-pore size paracellular model. Transcellular permeability, scaled by k(VF), was equated to that of Caco-2 at pH 6.5. The biophysical model predicted human jejunal permeability data within the experimental uncertainty. This investigation revealed several surprising predictions: (i) many molecules permeate predominantly (but not exclusively) by the paracellular route, (ii) the aqueous boundary layer thickness in the intestinal perfusion experiments is larger than expected, (iii) the mucosal surface area in awake humans is apparently nearly entirely accessible to drug absorption, and (iv) the relative "leakiness" of the human jejunum is not so different from that observed in a number of published Caco-2 studies.
Journal of Medicinal Chemistry 04/2010; 53(9):3566-84. · 5.61 Impact Factor
[show abstract][hide abstract] ABSTRACT: The permeability characteristics of 33 amphoteric drugs (about 64% zwitterions at physiological pH) were studied using the parallel artificial membrane permeability assay (PAMPA) at pH 6.5. The PAMPA data were modified to include the paracellular permeability component found in cellular monolayers based on a newly generalized version of a popular model devised for Caco-2 cells. These "in combo" PAMPA data were used to predict the human absolute bioavailability of the ampholytes. The analysis produced a good fit, with only five outliers whose transport properties, could be rationalized by (a) nonpassive permeability processes, (b) metabolic instability, and (c) the possible sensitivity to microclimate pH effects in the case of acidic ampholytes. With the exception of two compounds, all of the ampholytes with bioavailability <50% were predominantly transported by the paracellular route, surprisingly with several of the compounds having molecular weights exceeding 350 Da.
Journal of Medicinal Chemistry 11/2009; 53(1):392-401. · 5.61 Impact Factor
[show abstract][hide abstract] ABSTRACT: This paper reports on the magnetic properties of macroscopic colloidal crystals comprising silica-encapsulated FePt nanoparticles which assembled into three dimensional regular arrays by magnetic crystallization. The crystals have potential for magnetic separation of paramagnetic chemical or biochemical entities (e.g., proteins) by providing local magnetic fields at the interstitial sites to promote magnetic retention of the entities of interest. Structural characterizations by transmission electron microscopy and small angle x-ray scattering using a synchrotron source indicate a hcp arrangement of the nanoparticles in the colloidal crystals. The crystals have a large surface area (110.6 m <sup>2</sup> g <sup>-1</sup>) and a sharp pore size of 15 nm which is adjustable by controlling the silica shell thickness. Under applied fields, the colloidal crystals display substantially stronger magnetic responses than the unaligned particle assembly, thus favoring removal of the colloidal crystals from the mixture by magnetic means after magnetic separation. The potential of the colloidal crystals for separating paramagnetic biological or chemical species is demonstrated in absorption experiments using two proteins with and without a paramagnetic center.
Journal of Applied Physics 05/2009; · 2.21 Impact Factor
[show abstract][hide abstract] ABSTRACT: The development of a novel series of imidazole pyrimidine amides as cyclin-dependent kinase (CDK) inhibitors is described. Optimisation of inhibitory potency against multiple CDK's (1, 2 and 9) resulted in imidazole pyrimidine amides with potent in vitro anti-proliferative effects against a range of cancer cell lines. Excellent physiochemical properties and large margins against inhibition of CYP isoforms and the hERG ion channel were achieved by modification of lipophilicity and amine basicity. A candidate with disease model activity in human cancer cell line xenografts and with suitable physiochemical and pharmacokinetic profiles for intravenous (i.v.) dosing was selected for further development as AZD5597.
[show abstract][hide abstract] ABSTRACT: Colloid stable magnetic iron oxide nanoparticles, which undergo reversible precipitation from aqueous solution with external magnetic flux, can have many potential applications. However, the lack of generic homogeneous anchoring sites on a magnetic nanoparticle surface for binding of chemical/biochemical species under a wide range of conditions is one key problem. It is shown that a small size iron oxide nanoparticle encapsulated in a thin silica shell can offer specific sites to bind protein molecules via surface silanol groups electrostatically at pH 7.4 without severe denaturing of the bulky protein structure. As a result, we show that a high loading of bovine serum albumin (BSA) of 85 mg/g can be anchored on the silica-encapsulated iron oxide. FTIR, circular dichroism, and binding constant (using site I and site II drugs) measurements show only a small degree of conformational alteration upon immobilization. A partial unfolding of secondary structures on the external sheath of the protein due to competitive hydrogen bonding interactions of functional groups such as −CO and −NH with surface acidic hydroxyl groups is shown to take place despite the use of buffered pH 7.4 solution. In contrast to the blockage of drug binding sites reported in the case of anchored BSA on extended silica surface, our results clearly show that the internal hydrophobic sites I and II of the immobilized BSA on this silica-based magnetic nanoparticle remain intact for drugs binding at a high degree.
Journal of Physical Chemistry C - J PHYS CHEM C. 12/2008; 113(2).
[show abstract][hide abstract] ABSTRACT: An in situ analytical approach to the measurement of supported liquid membrane permeability is reported. The method consists of a spectrophotometric method to measure transport through a membrane-supported lipid solution, using a rotating-diffusion cell configuration to overcome limits arising from transport through the aqueous solution boundary layer in stationary systems. Rotation frequencies are almost two orders of magnitude higher than those employed previously for rotating-diffusion studies of membrane transport. The method is illustrated with the transport of warfarin [1-(4'-hydroxy-3'-coumarinyl)-1-phenyl-3-butanone]. The use of the rotating-diffusion approach permits accurate calculation of the aqueous phase boundary layer thickness, which has hitherto been treated as an adjustable parameter in studies of membrane permeability. Further, it is shown that the analyte diffusion coefficient can be determined readily using liquid-liquid electrochemistry.
The Analyst 06/2008; 133(5):655-9. · 4.23 Impact Factor
[show abstract][hide abstract] ABSTRACT: Recent development in assembling nanoparticles as building blocks into macroscopic functional structures or devices to harness the size-dependent properties of individual particles is an exciting new direction. Here, it is demonstrated that well-dispersed silica- encapsulated superparamagnetic FePt nanoparticles in solution can be assembled into high-quality, needle- or rod-shaped solid-state supercrystals by applying inhomogeneous external magnetic field together with controlled evaporation. By use of an aggregation model based on the population balance equation technique, this charactersitic morphology of the magnetic asembled crystals can be successfully derived.
Journal of Physical Chemistry C - J PHYS CHEM C. 04/2008; 112(20).