Phosphine boranes as less hydrophobic building blocks than alkanes and silanes: Structure-property relationship and estrogen-receptor-modulating potency of 4-phosphinophenol derivatives

Increasing structural options in medicinal chemistry is important for the development of novel and distinctive drug candidates. In this study, we focused on phosphorus-containing functionalities. We designed and synthesized a series of phosphinophenol derivatives and determined their physicochemical properties, including hydrophobicity parameter LogP, and their biological activity toward estrogen receptor (ER). Notably, the phosphine borane derivatives (9 and 14) exhibited potent ER-antagonistic activity, exceeding the potency of the corresponding alkane (15) and silane (16) derivatives, despite having a less hydrophobic nature. The determined physicochemical parameters will be helpful for the rational design of phosphorus-containing biologically active compounds. Our results indicate that phosphine boranes are a promising new chemical entry in the range of structural options for drug discovery.     

The Influence of Rhamnolipids on Aliphatic Fractions of Diesel Oil Biodegradation by Microorganism Combinations

Twelve different bacteria–yeast combinations were tested for determination of their ability to biodegrade diesel oil. The cell surface properties of the bacterial and yeast strains were correlated with the type of carbon source used in the experiments. The highest biodegradation of diesel oil after 7 days was obtained for the following combinations: Aeromonas hydrophila MR4–Yarrowia lipolytica EH 56 (87 %) and Xantomonas maltophila MRP7–Candida maltosa EH15 (90 %). Degradation performances of 10 of 12 combinations were enhanced by the presence of rhamnolipids. The highest increases were observed for A. hydrophila MR4–C. maltosa EH15 (from 34 to 67 %), A. hydrophila MR4–C. maltosa EH60 (from 47 to 76 %) and for Pseudomonas stutzeri MR7–C. maltosa EH60 (from 29 to 79 %). The addition of rhamnolipids to the system reduces the removal time of diesel oil from the contaminated water and changes the microbial adhesion to hydrocarbons. Modification of the cell surface of the tested strain during biodegradation is a very important factor determining the removal of hydrophobic compounds.     

Pervaporation Aided Esterification of Carboxylic Acids with Ethanol Catalyzed by Porcine pancreatic Lipase

The results of pervaporation-coupled esterification of various carboxylic acids with ethanol catalyzed by Porcine pancreatic lipase are reported. The effect of lipase and substrate concentrations has been studied and the advantage of pervaporation on the equilibrium conversion has been deduced. The kinetics of reaction were analyzed with a three-parameter model which coupled the effect of pervaporation. The intrinsic kinetic constants for all the reactions were estimated and correlated with the carbon number, an indicator of hydrophobicity of the acids. It was found that the rate constant increases with decrease in carbon number. The experimental concentration profiles were simulated from the model for all the reactions and the model prediction was found to be reasonably good. The water permeability was also correlated well with acid hydrophobicity. The pervaporation coupled reaction efficiency, as represented by the reaction time for equilibrium conversion, was found to bear a profound relation to membrane surface area per unit volume of the reaction mixture (A/V). The time for equilibrium conversion was found to decrease with an increase in A/V value, reaching a minimum and then increasing with a further increase of A/V. A probable explanation has been postulated for such an observation.     

Systematic research on the pretreatment of peptides for quantitative proteomics using a C18 microcolumn

Reversed phase microcolumns have been widely used for peptide pretreatment to desalt and remove interferences before tandem LC–MS in proteomics studies. However, few studies have characterized the effects of experimental parameters as well as column characteristics on the composition of identified peptides. In this study, several parameters including the concentration of ACN in washing buffer, the microcolumn's purification effect, the peptide recovery rate, and the dynamic‐binding capacity were characterized in detail, based upon stable isotope labeling by amino acids in a cell culture quantitative approach. The results showed that peptide losses can be reduced with low ACN concentration in washing buffers resulting in a recovery rate of approximately 82%. Furthermore, the effects of ACN concentration and loading amount on the properties of identified peptides were also evaluated. We found that the dynamic‐binding capacity of the column was approximately 26 μg. With increased loading amounts, more hydrophilic peptides were replaced by hydrophobic peptides.     

Engineering surface hydrophobicity improves activity of Bacillus thermocatenulatus lipase 2 enzyme

Bacillus thermocatenulatus lipase 2 (BTL2) is a promising industrial enzyme used in biodiesel production. Although BTL2 has high thermostability and good resistance to organic solvents, the activity of BTL2 is suboptimal for industrial processes. To improve BTL2 activity, we engineered BTL2 lipase by modulating hydrophobicity of its lid domain. Through site‐directed mutagenesis, we constructed three mutants, namely Y225F+S232A, S232A+T236V and Q185L, to cover all uncharged hydrophilic amino acids within the lid domain. Activities of these mutants were characterized. Our findings suggest that one mutant (Y225F+S232A) showed ～35% activity increase in catalyzing heterogeneous hydrolytic reactions relevant for industrial applications. A mathematical framework was established to account for different molecular events that contribute to the observed apparent catalytic activities. Increases in hydrophobicity of lid domains were associated with increased interfacial adsorption of lipases and lower molecular enzymatic activities. The measured apparent activities of lipases include contributions from both events. Lid hydrophobicity can thus result in different changes in lipase activities depending on the mutation site. Our work demonstrates the feasibility of increasing BTL2 activity by modulating the hydrophobicity of lid domains and provides some guidelines for further improving BTL2 activity.     

Significance of the class II hydrophobin FgHyd5p for the life cycle of Fusarium graminearum

Hydrophobins are small secreted proteins ubiquitously found in filamentous fungi. Some hydrophobins were shown to have functions in fungal development, while others lack known function. Class II hydrophobins from Fusarium graminearum and Fusarium culmorum are characterized by formation of low stability aggregates and their solubility in organic solvents. They are economically relevant to the brewing industry because they can induce beer gushing. Since cellular functions of Hyd5p's are still unknown, we analyzed the influence of FgHyd5p on growth and morphology of F. graminearum using FgΔhyd5 knock-out mutants expressing sGFP under the control of the hyd5 promoter and compared them with the performance of the parent wild type strain. Results demonstrate that FgHyd5p does not affect the colony and hyphal morphology. FgHyd5p affects the hydrophobicity of aerial mycelia but had no obvious function in penetration of hyphae through the water air interface. The hydrophobin affects the morphology of conidia, but not their fitness. Different sources of carbon and nitrogen as well as different pH have no effect on the expression of the hyd5 gene, which was demonstrated to be expressed upon growth of F. graminearum on hydrophobic surfaces.     

Rho-associated kinase (ROCK) inhibition reverses low cell activity on hydrophobic surfaces

Hydrophobic polymers do not offer an adequate scaffold surface for cells to attach, migrate, proliferate, and differentiate. Thus, hydrophobic scaffolds for tissue engineering have traditionally been physicochemically modified to enhance cellular activity. However, modifying the surface by chemical or physical treatment requires supplementary engineering procedures. In the present study, regulation of a cell signal transduction pathway reversed the low cellular activity on a hydrophobic surface without surface modification. Inhibition of Rho-associated kinase (ROCK) by Y-27632 markedly enhanced adhesion, migration, and proliferation of osteoblastic cells cultured on a hydrophobic polystyrene surface. ROCK inhibition regulated cell-cycle-related molecules on the hydrophobic surface. This inhibition also decreased expression of the inhibitors of cyclin-dependent kinases such as p21^cip1 and p27^kip1 and increased expression of cyclin A and D. These results indicate that defective cellular activity on the hydrophobic surface can be reversed by the control of a cell signal transduction pathway without physicochemical surface modification.     

Reduced curvilinear velocity of boar sperm on substrates with increased hydrophobicity

The curvilinear velocity (VCL) of boar spermatozoa between standard microscopy glassware decreases when the slides are coated with the hydrophobic polymer polystyrene (PS) compared with the less hydrophobic poly(methyl methacrylate) (PMMA) coating. Sperm from three boars were observed and analyzed using particle tracking software. The VCL did not differ significantly between coatings of different thickness, indicating no penetration of the sperm into the coating and that only the surface layer of the polymer film interacts with the sperm and buffer medium. The VCL of sperm between PS-coated surfaces was significantly reduced compared with PMMA surfaces (P < 0.0001), and this was attributed to a stronger hydrophobic effect between PS and water. The size of this effect varied between different boars, perhaps as a consequence of variations in hydrophobicity of sperm from different boars or different ejaculates. The modification of surface properties in this way may improve our understanding of sperm behavior and may provide improvements to assisted conception techniques as animal or human sperm used in assisted conception are frequently manipulated in laboratory plastics as part of diagnostic procedures (e.g., semen analysis) or before injection into an oocyte or during the co-incubation with the oocyte in IVF. Controlling the velocity of sperm using the interaction properties of inert polymer coatings could lead to new sperm selection procedures for clinical use or the development of model systems to better understand sperm–surface interactions.     

Development of a potent and orally active activator of adenosine monophosphate-activated protein kinase (AMPK), ASP4132, as a clinical candidate for the treatment of human cancer

Adenosine monophosphate (AMP)-activated protein kinase (AMPK) plays a key role in maintaining cellular metabolism. AMP or adenosine diphosphate (ADP) levels rise during metabolic stress, such as during nutrient starvation, hypoxia and muscle contraction, and bind to AMPK to induce activity. Recently, activation of AMPK has been considered an attractive therapeutic strategy in the field of human oncology. Structural optimization of lead compound 2, a new type of AMPK activator with potent AMPK activation activity and attractive selective growth inhibition against human cancer cells, improved aqueous solubility, metabolic stability and animal pharmacokinetics (PK) and culminated in the identification of (5-{1-[(6-methoxypyridin-3-yl)methyl]piperidin-4-yl}-1H-benzimidazol-2-yl)(4-{[4-(trifluoromethyl)phenyl]methyl}piperazin-1-yl)methanone ditosylate, ASP4132 (28). Studies on ASP4132 had advanced to clinical trials for the treatment of cancer.     

Effect of hydrophobicity degree of polymer particles on lipase immobilization and on biocatalyst performance

Abstract

Many studies describe the advantages of using hydrophobic particles on lipase immobilisation. However, many of these works neglect the effect of other variables of the supports, such as specific area and porosity, on the biocatalyst performance, and do not evaluate the influence of the hydrophobicity level of the particles on the biocatalysts’ activity as a single variable. Thus, the focus of the present work was to evaluate the effect of the hydrophobicity degree of polymeric particles on the biocatalysts’ activities, mitigating the influence of other variables. The study was divided into two steps. Firstly, distinct particles, exhibiting different composition and hydrophobicity levels, were used for the immobilization of a commercial lipase B from Candida antarctica (CAL-B). Then, distinct core-shell polymeric particles presenting different functional compounds on the surface were produced, using as comonomers styrene, divinylbenzene, 1-octene, vinylbenzoate and cardanol. Such particles were subsequently used for CAL-B immobilisation and the performance of the biocatalysts was evaluated on hydrolysis (using p-nitrophenyl laurate, as substrate) and esterification (using ethanol and oleic acid, as substrate) reactions. Based on the screening step, it was observed that for non-porous particles the correlation coefficients between the hydrophobicity level of the supports and the biocatalysts performance, for both hydrolysis and esterification reactions, were very low (0.32 and 0.45, respectively). It highlights that there was no significant correlation between these variables and that, probably, the chemical composition of the polymeric chains affects more significantly the biocatalyst performance. Then, analysing the subsequent stage, it was observed that small changes in the surface composition of the core-shell particles result in significant changes on the textural properties of the supports (specific area ranging from 1.2?m².g^?1 to 18.3?m².g^?1; and contact angles ranging from 71° (hydrophilic particles) to 92° (hydrophobic supports) when polymer films were put in contact with water). Such particles were also employed on CAL-B immobilization and it was noticed that higher correlation coefficients were achieved for hydrolysis (ρ?=?0.53) and esterification (ρ?=?0.74) reactions. Therefore, it is shown that the hydrophobicity degree of such supports starts to affect more effectively the biocatalysts performance when other textural features of the supports become more significant, such as specific area and porosity.