Pig liver esterases PLE1 and PLE6: heterologous expression, hydrolysis of common antibiotics and pharmacological consequences.

Carboxylesterases, historically referred as non-specific esterases, are ubiquitous hydrolases with high catalytic efficiency. Without exceptions, all mammalian species studied contain multiple forms of carboxylesterases. While having been widely studied in humans and experimental animals, these enzymes remain to be characterized in farm animals. In this study, we showed that pig liver esterase 1 (PLE1) and pig liver esterase 6 (PLE6) were highly active toward amoxicillin (AMO) and ampicillin (AMP), two major antibiotics that are widely used in food-supplements. Mass-spectrometric analysis established that the hydrolysis occurred at the ß-lactam amide bond and the hydrolysis drastically decreased or completely eliminated the antibacterial activity. Furthermore, hydrolytic activity and proteomic analysis suggested that trace PLEs existed in pig plasma and contributed little to the hydrolysis of AMO and AMP. These results suggested that carboxylesterases-based hydrolysis determines the therapeutic intensity of these and related antibiotics and the magnitude of the determination occurs in a species-dependent manner.

Bactericidal activity-tunable conjugated polymers as a human-friendly bactericide for the treatment of wound infections.

Photodynamic therapy (PDT) has been reported to be an effective alternative to combat bacterial infections even those triggered by drug-resistant strains as there is little chance to develop resistance to this therapy. Therefore, it is imperative to design and synthesize a superior photo-active bactericide for the treatment of bacterial infections. Herein, we synthesized three bactericidal activity-tunable conjugated polymers (P1-P3) with various photoactive capabilities and employed them for the treatment of wound infections with little damage to cells; by altering the construction unit of π-conjugated backbone structures with electron-rich and electron-deficient aromatic heterocycles, the optical properties and ability of reactive oxygen species (ROS) generation could be regulated; this resulted in a tunable killing ability. The cationic quaternary ammonium (QA) groups on the side chains endowed the CPs with not only good dispersibility but also a better interaction with the negatively charged membrane of bacteria. The antibacterial experiments towards ampicillin-resistant Escherichia coli TOP10 (E. coli) and the treatment of wound infections in mice indicate that the P1-P3 have varied bactericidal activities; moreover, P3 has been demonstrated to be a human-friendly bactericide with excellent antibacterial capability. It not only acts as a potential bactericide for the practical treatment of infectious wounds, but also offers guidance for the design and structure control of photo-active bactericides.

[Discovery of Novel Biologically Active Compounds of Natural Origin, with a Focus on Anti-tumor Activity].

Numerous clinically valuable medicines, including anticancer drugs, have been developed from biologically active natural compounds and their structurally related derivatives. This review discusses novel natural compounds with promising biological activities and those with novel chemical structures. Glaziovianin A, an isoflavone isolated from the leaves of Ateleia glazioviana (Legminosae), inhibited cell cycle progression at the M-phase with an abnormal spindle structure. AU-1 and YG-1, 5ß-steroidal glycosides isolated from the whole plants of Agave utahensis and the underground parts of Yucca glauca (Agavaceae), induced apoptosis of HL-60 cells via caspase-3 activation. Lycolicidinol, an alkaloid isolated from the bulbs of Lycoris albiflora (Amaryllidaceae), induced transient autophagy and morphological changes in mitochondria in the early stage of the apoptotic cell death process in HSC-2 cells. Taccasterosides isolated from the rhizomes of Tacca chantrieri (Taccaceae) and stryphnosides isolated from the pericarps of Stryphnodendron fissuratum (Legminosae) are steroidal and triterpene glycosides with unique chemical structures having novel sugar sequences.

Evaluation of solubility and partition properties of ampicillin-based ionic liquids.

In order to overcome the problems associated with low water solubility, and consequently low bioavailability of active pharmaceutical ingredients (APIs), herein we explore a modular ionic liquid synthetic strategy for improved APIs. Ionic liquids containing L-ampicillin as active pharmaceutical ingredient anion were prepared using the methodology developed in our previous work, using organic cations selected from substituted ammonium, phosphonium, pyridinium and methylimidazolium salts, with the intent of enhancing the solubility and bioavailability of L-ampicillin forms. In order to evaluate important properties of the synthesized API-ILs, the water solubility at 25 °C and 37 °C (body temperature) as well as octanol-water partition coefficients (Kow's) and HDPC micelles partition at 25 °C were measured. Critical micelle concentrations (CMC's) in water at 25 °C and 37 °C of the pharmaceutical ionic liquids bearing cations with surfactant properties were also determined from ionic conductivity measurements.

Experimental determination of ampicillin adsorption to nanometer-size Al2O3 in water.

Transport of antibiotics in soil-water systems is controlled in part by adsorption to nanometer-size (10(-9)m) particles. Batch adsorption experiments were performed with ampicillin, a common amphoteric antibiotic, and 50 nm-Al(2)O(3) (alpha-alumina) at different pH conditions. Sorption to Al(2)O(3) can be described by linear isotherms for 2.9 microM-2.9 mM ampicillin concentrations. Distribution coefficients (K(d)) are 11.1 (+/-0.32)L kg(-1) at pH 2, 0.55 (+/-.04) L kg(-1) at pH 4, 21.9 (+/-0.9) L kg(-1) at pH 6, and 39.5 (+/-2.2) L kg(-1) at pH 8. At pH 2, approximately 47% of the initially adsorbed drug was removable by rinsing, at pH 4-56% was removed. Only 7% of the drug could be removed by rinsing at pH 6, and 3% at pH 8. Weak electrostatic forces dominate at pH<4, and stronger attachment mechanisms at higher pH. Low yields in rinsing (desorption) experiments at pH6 indicate strong attachment mechanisms, either electrostatic or possibly surface complexation.

Chemical enhancement of torsinA function in cell and animal models of torsion dystonia.

Movement disorders represent a significant societal burden for which therapeutic options are limited and focused on treating disease symptomality. Early-onset torsion dystonia (EOTD) is one such disorder characterized by sustained and involuntary muscle contractions that frequently cause repetitive movements or abnormal postures. Transmitted in an autosomal dominant manner with reduced penetrance, EOTD is caused in most cases by the deletion of a glutamic acid (DeltaE) in the DYT1 (also known as TOR1A) gene product, torsinA. Although some patients respond well to anticholingerics, therapy is primarily limited to either neurosurgery or chemodenervation. As mutant torsinA (DeltaE) expression results in decreased torsinA function, therapeutic strategies directed toward enhancement of wild-type (WT) torsinA activity in patients who are heterozygous for mutant DYT1 may restore normal cellular functionality. Here, we report results from the first-ever screen for candidate small molecule therapeutics for EOTD, using multiple activity-based readouts for torsinA function in Caenorhabditis elegans, subsequent validation in human DYT1 patient fibroblasts, and behavioral rescue in a mouse model of DYT1 dystonia. We exploited the nematode to rapidly discern chemical effectors of torsinA and identified two classes of antibiotics, quinolones and aminopenicillins, which enhance WT torsinA activity in two separate in vivo assays. Representative molecules were assayed in EOTD patient fibroblasts for improvements in torsinA-dependent secretory function, which was improved significantly by ampicillin. Furthermore, a behavioral defect associated with an EOTD mouse knock-in model was also rescued following administration of ampicillin. These combined data indicate that specific small molecules that enhance torsinA activity represent a promising new approach toward therapeutic development for EOTD, and potentially for other diseases involving the processing of mutant proteins.

Formulation development and in vitro and in vivo evaluation of membrane-moderated transdermal systems of ampicillin sodium in ethanol: pH 4.7 buffer solvent system.

The objective of the present study was to develop membrane-moderated transdermal systems of ampicillin sodium and to evaluate them with respect to various in vitro and in vivo parameters. The membrane-type transdermal systems were prepared using a drug with various antinucleant polymers-hydroxypropyl methylcellulose (HPMC), methylcellulose (MC), cellulose acetate phthalate, chitosan, sodium alginate (SA), and sodium carboxymethylcellulose-in an ethanol: pH 4.7 buffer volatile system by the solvent evaporation technique with HPMC as the rate-controlling membrane for all the systems. The swelling properties of the polymers were studied, and drug-polymer interaction studies were performed. The patches were subjected to various physicochemical studies, in vitro release studies, permeation studies, and skin irritation studies. The best patch among the formulations was selected for further in vivo studies. Compared to the other patches, SA exhibited the highest moisture content at 16%; a 21% moisture uptake was found with MC. The release and permeation of the drug from the SA patch was found to be the maximum. The in vivo study of the SA patch exhibited a peak plasma concentration C(max) of 126 microg/mL at T(max) 4 hours. Hence, it can be concluded that hydrophilic ampicillin sodium can be developed as a transdermal delivery system with SA that is an alternative to intravenous administration and has minimal adverse effects.

Channel activity of OmpF monitored in nano-BLMs.

Free-standing lipid bilayer membranes can be formed on small apertures (60 nm diameter) on highly ordered porous alumina substrates. The formation process of the membranes on a 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol submonolayer was followed by impedance spectroscopy. After lipid bilayers had thinned, the reconstitution and ionic conducting properties of the outer membrane protein OmpF of E. coli were monitored using single-channel recordings. The characteristic conductance states of the three monomers, fast kinetics, and subconductance states were observed. Blockade of the ion flow as a result of interaction of the antibiotic ampicillin with the protein was verified, indicating the full functionality of the protein channel in nanometer-scale bilayer membranes.

Microcalorimetric evaluation of the in vitro compatibility of amoxicillin/clavulanic acid and ampicillin/sulbactam with ciprofloxacin.

Solution calorimetric technique has been used to determine the compatibility of binary and ternary systems of ampicillin trihydrate (AMP), sulbactam sodium (SS), amoxicillin trihydrate (AM), potassium clavulanate (PC) and ciprofloxacin hydrochloride (CP). The enthalpy of solution (DeltasolH) were obtained over a wide range of composition in the pH range 2-9. For all the pure drugs the DeltasolH is endothermic in nature. The molar enthalpies of interaction of binary (DeltaHbi.E) and ternary (DeltaHter.E) mixtures of the drugs in aqueous buffers have been determined. The DeltaHbi.E for all binary systems is negative and pH dependent (maximum pH 6-8) indicating the interaction among charged species of the drugs. In case of binary systems with CP the magnitude of DeltaHbi.E indicate strong interactions. The variation and magnitude of DeltaHbi.E for the systems is discussed in terms of hydrogen bonding and van der Waal's interaction in the solution. The interaction parameter for ternary systems (A) is positive indicating repulsive interaction among the drugs. The coefficients hi's calculated from Redlich-Kister equation for binary systems (DeltaHbi.E) and ternary interaction parameter (A) were used to predict the compatibility of the marketed formulations in pH range studied.

Penicillin derivatives induce chemical structure-dependent root development, and application for plant transformation.

We investigated five penicillin derivatives that are popularly used for transformation experiments with Agrobacterium rhizogenes-penicillin G, carbenicillin, ampicillin, amoxicillin and cephalexin-for their effects on the growth and morphology of Beta vulgaris, Capsicum annuum and Glehnia littoralis roots. Attention was given to the relationship between their chemical structures and functions. Ampicillin was found to stimulate root elongation but inhibit root branching, whereas carbenicillin inhibited root elongation but promoted root branching. Root cultures were also exposed to hydrolyzed products of these antibiotics-i.e. phenylmalonic acid (PM), phenylglycine and 6-aminopenicillanic acid (6-APA): PM inhibited root elongation the most, while root elongation was supported best by 6-APA. These results indicate that both the side chains and the major component of penicillin derivatives affect root development and that the nature of the side chains is responsible for the responses. Ampicillin but not carbenicillin was used in subsequent experiments described herein to eliminate bacteria and to support root growth of transformants of the recalcitrant plants.

Effects of oils and pharmaceutical excipients on the bioavailability of ampicillin orally administered, different oily and aqueous suspensions in rabbit.

The in vivo bioavailability and in vitro drug-release studies of ampicillin trihydrate in different oily and aqueous suspensions have been investigated. In addition, partition, solubility, and rheological measurements have also been carried out. The in vivo experimental design was based on a 6 x 6 latin square using the rabbit as the test animal. The bioavailability of ampicillin was determined using the plasma levels, which were measured microbiologically. Results of the study showed that oily and sucrose-containing aqueous formulations enhanced the extent of ampicillin absorption, although not statistically significantly, but was close to the borderline of significance. Ampicillin appears to be absorbed at essentially the same rate from both aqueous and oily formulations. The latter showed plasma-level time curves with biphasic absorption and are likely to produce prolonged plasma concentrations of ampicillin because of the effects of enterohepatic recycling. Viscosity appears to play an insignificant role in the results obtained since the bioavailability parameters correlate poorly with the viscosity except Cmax. It is suggested that enhancement in the bioavailability of ampicillin is due to the decrease in the gut transit rate brought about by the oil which predominates and masks the other effects of viscosity and osmotic effects of sucrose. The existence of a correlation between the in vitro drug-release rate (t50%) and viscosity and the lack of a correlation between in vivo and in vitro parameters support the above suggestion and indicate that traditional dissolution rate tests, such as flask-stirrer method, are unsatisfactory as bioavailability indicators when applied to dosage forms that caused marked changes in physiological factors like GER and biliary excretion.