Caffeic Acid Phosphanium Derivatives: Potential Selective Antitumor, Antimicrobial and Antiprotozoal Agents.

Caffeic acid (CA) is one of the most abundant natural compounds present in plants and has a broad spectrum of beneficial pharmacological activities. However, in some cases, synthetic derivation of original molecules can expand their scope. This study focuses on the synthesis of caffeic acid phosphanium derivatives with the ambition of increasing their biological activities. Four caffeic acid phosphanium salts (CAPs) were synthesized and tested for their cytotoxic, antibacterial, antifungal, and amoebicidal activity in vitro, with the aim of identifying the best area for their medicinal use. CAPs exhibited significantly stronger cytotoxic activity against tested cell lines (HeLa, HCT116, MDA-MB-231 MCF-7, A2058, PANC-1, Jurkat) in comparison to caffeic acid. Focusing on Jurkat cells (human leukemic T cell lymphoma), the IC50 value of CAPs ranged from 0.9 to 8.5 µM while IC50 of CA was >300 µM. Antimicrobial testing also confirmed significantly higher activity of CAPs against selected microbes in comparison to CA, especially for Gram-positive bacteria (MIC 13-57 µM) and the yeast Candida albicans (MIC 13-57 µM). The anti-Acanthamoeba activity was studied against two pathogenic Acanthamoeba strains. In the case of A. lugdunensis, all CAPs revealed a stronger inhibitory effect (EC50 74-3125 µM) than CA (>105 µM), while in A. quina strain, the higher inhibition was observed for three derivatives (EC50 44-291 µM). The newly synthesized quaternary phosphanium salts of caffeic acid exhibited selective antitumor action and appeared to be promising antimicrobial agents for topical application, as well as potential molecules for further research.

Efficacy of Pythium oligandrum on improvement of lucerne yield, root development and disease score under field conditions.

Introduction: Biological control of root diseases of lucerne (Medicago sativa L.) has potential benefits for stand performance but this remains unsupported by evidence from practical field studies. Methods: In field experiments at three sites our objectives were to determine the effect of Pythium oligandrum, as spring, autumn and intensive regime treatments on (i) lucerne plant density and root traits development, and (ii) forage yield and forage traits. Lucerne stands were managed under two or three treatments: non-treated control and P. oligandrum applied at two intensities of application under four-cut utilization. Results and discussion: Under relatively dry conditions (annual mean 10°C and <500 mm precipitation) lucerne dry matter yield was significantly reduced by 6%, which could be related to mechanisms of inappropriate stimulation and disturbance of the balance between auxins and ethylene. Under annual precipitation of >500 mm, positive impacts on stand height or fine root mass were observed for the autumn and intensive treatments where positive root response was visible only in alluvial soil. However, these changes did not result in higher yield and probably more applications per year will be needed for significant forage yield improvement. This study highlights the limits of field-scale biological control in which the potential of P. oligandrum for lucerne productivity improvement was realised only under a humid environment or deep alluvial soils, where higher root disease infestation may also be expected.

Silver Nanoparticles Stabilized with Phosphorus-Containing Heterocyclic Surfactants: Synthesis, Physico-Chemical Properties, and Biological Activity Determination.

Phosphorus-containing heterocyclic cationic surfactants alkyldimethylphenylphospholium bromides with the alkyl chain length 14 to 18 carbon atoms were used for the stabilization of silver nanodispersions. Zeta potential of silver nanodispersions ranges from +35 to +70 mV, which indicates the formation of stable silver nanoparticles (AgNPs). Long-chain heptadecyl and octadecyl homologs of the surfactants series provided the most intensive stabilizing effect to AgNPs, resulting in high positive zeta potential values and smaller diameter of AgNPs in the range 50-60 nm. A comparison with non-heterocyclic alkyltrimethylphosphonium surfactants of the same alkyl chain length showed better stability and more positive zeta potential values for silver nanodispersions stabilized with heterocyclic phospholium surfactants. Investigations of biological activity of phospholium-capped AgNPs are represented by the studies of antimicrobial activity and cytotoxicity. While cytotoxicity results revealed an increased level of HepG2 cell growth inhibition as compared with the cytotoxicity level of silver-free surfactant solutions, no enhanced antimicrobial action of phospholium-capped AgNPs against microbial pathogens was observed. The comparison of cytotoxicity of AgNPs stabilized with various non-heterocyclic ammonium and phosphonium surfactants shows that AgNPs capped with heterocyclic alkyldimethylphenylphospholium and non-heterocyclic triphenyl-substituted phosphonium surfactants have the highest cytotoxicity among silver nanodispersions stabilized by the series of ammonium and phosphonium surfactants.

Synthesis, physicochemical properties and biological activities of novel alkylphosphocholines with foscarnet moiety.

A series of alkylphosphocholines with foscarnet moiety was synthesized. The structure of these zwitterionic amphiphiles was modified in both polar and non-polar parts of surfactant molecule. Investigations of physicochemical properties are represented by the determination of critical micelle concentration, the surface tension value at the cmc and the surface area per surfactant head group utilising surface tension measurements. Hydrodynamic diameter of surfactant micelles was determined using the dynamic light scattering technique. Alkylphosphocholines exhibit significant cytotoxic, anticandidal (Candida albicans) and antiamoebal (Acanthamoeba spp. T4 genotype) activity. The relationship between the structure, physicochemical properties and biological activity of the tested compounds revealed that lipophilicity has a significant influence on biological activity of the investigated surfactants. More lipophilic alkylphosphocholines with octadecyl chains show cytotoxic activity against cancer cells which is higher than that of the compounds with shorter alkyl chains. The opposite situation was observed in case of anticandidal and antiamoebal activity of these surfactants. The most active compounds were found to have pentadecyl chains. The foscarnet analogue of miltefosine C15-PFA-C showed the highest anticandidal activity. The minimum value of anticandidal activity of this compound is 1,4 µM thus representing the highest anticandidal activity found within the group of alkylphosphocholines.

The Synthesis, Self-Assembled Structures, and Microbicidal Activity of Cationic Gemini Surfactants with Branched Tridecyl Chains.

Cationic gemini surfactants with polymethylene spacer and linear alkyl chains containing an even number of carbon atoms have been extensively studied in the recent past, with the emphasis put on the determination of their aggregation behaviour in aqueous solution and their biological properties. However, the information on the aggregation of branched gemini surfactants with an odd number of carbon atoms in their alkyl chains is only sparsely reported in the literature. To help cover this gap in the research of cationic gemini surfactants, a series of branched bisammonium cationic gemini surfactants with an odd number of carbon atoms in alkyl chains (tridecane-2-yl chains) and a polymethylene spacer with a variable length ranging from 3 to 12 carbon atoms have been synthesized and investigated. Critical micelle concentration, which was determined by three methods, was found to be in the order 10-4 mol/L. A comparison of the obtained data of the novel series of tridecyl chain geminis with those of gemini surfactants with dodecyl chains and an identical spacer structure revealed that structural differences between both series of gemini surfactants result in different aggregation and surface properties for surfactants with 6 and 8 methylene groups in the spacer (N,N'-bis(tridecane-2-yl)-N,N,N',N'-tetramethylhexane-1,6-diaminium dibromide and N,N'-bis(tridecane-2-yl)-N,N,N',N'-tetramethyloctane-1,8-diaminium dibromide) with the cmc values 8.2 × 10-4 mol/L and 6.5 × 10-4 mol/L, respectively, as determined by surface tension measurements. Particle size analysis showed the formation of small stable spherical micelles in the interval between 2.8 and 5 nm and with zeta potential around +50 mV, which are independent of surfactant concentration and increase with the increasing spacer length. Microbicidal activity of 13-s-13 gemini surfactants was found to be efficient against Gram-positive, Gram-negative bacteria and yeast.

pH-Sensitive N,N-Dimethylalkane-1-amine N-Oxides in DNA Delivery: From Structure to Transfection Efficiency.

pH-sensitive liposomes composed of homologues of series of N,N-dimethylalkane-1-amine N-oxides (CnNO, n = 8-18, where n is the number of carbon atoms in the alkyl substituent) and neutral phospholipid dioleoylphosphatidylethanolamine (DOPE) were prepared at two molar ratios (CnNO/DOPE = 0.4:1 and 1:1) and tested for their in vitro transfection activity. Several techniques (SAXS/WAXS, UV-vis, zeta potential measurements, confocal microscopy) were applied to characterize the system in an effort to unravel the relationship among the transfection efficiency, structure, and composition of the lipoplexes. The transfection efficiency of CnNO/DOPE for plasmid DNA in U2OS cells follows a quasi-parabolic dependence on CnNO's alkyl substituent length with a maximum at n = 16. The transfection efficiency of CnNO/DOPE (n = 12-18) lipoplexes was found to be higher than that of commercially available Lipofectamine 2000. C16NO/DOPE also positively transfected HEK 293T and HeLa cells. Small-angle X-ray scattering (SAXS) shows large structural diversity depending on the complex's composition and pH. Transfection efficiencies mediated by two structures, either a condensed lamellar (Lαc) or epitaxially connected Lαc and a condensed inverted hexagonal (HIIc) phase (Lαc & HIIc), were found to be very similar. The change in pH from acidic to neutral induces phase transition Lαc & HIIc → QII + Lα, with cubic phase QII of the Pn3m space group. QII detected in lipoplexes of most efficient composition CnNO/DOPE (n = 16 and 18) facilitates DNA release and promotes its internalization in the cell.

Self-Assembly Properties of Cationic Gemini Surfactants with Biodegradable Groups in the Spacer.

: Self-assembly properties of cationic gemini surfactants with biodegradable amide or ester groups in the spacer were investigated utilising time-resolved fluorescence quenching, dynamic light scattering and zeta potential measurements. A correlation between aggregation parameters such as micelle aggregation number, micelle size and zeta potential with the structure of gemini molecules was made. For gemini molecules with medium spacer lengths, micelle aggregation number does not change much with the surfactant concentration. When the spacer is extended, a stronger aggregation tendency is observed for gemini surfactant molecules with two ester groups in the spacer and the aggregation number increases. The assumption of stronger aggregation of ester-based gemini molecules at larger spacer number values is also documented by measurements of the size and zeta potential of ester-based micelles. The explanation of the difference in aggregation ability of amide-based and ester-based gemini molecules is related to the structural features of gemini molecules, notably to the larger flexibility and denser arrangement of ester-based gemini molecules in a micelle. To support this assumption, optimised 3D models of the studied gemini molecules were constructed. Correspondingly, the calculations show smaller size and interfacial area for ester-based gemini conformers.

Improvement of Glibenclamide Water Solubility by Nanoparticle Preparation.

Glibenclamide, a drug used for the treatment of type 2 diabetes, belongs to Class II of Biopharmaceutical Classification System. It is a highly permeable, but poorly water-soluble drug. Nanoparticles of glibenclamide were prepared by an emulsion solvent evaporation method using dichloromethane as a solvent of glibenclamide and 3% (w/w) aqueous solution of carboxymethyl dextran sodium salt as a stabilizer, which was found as optimal. A solubility test comparing the water solubility of glibenclamide bulk and nanoparticles confirmed the improved (2-fold higher) solubility of glibenclamide nanoparticles (0.045 µg/ml) compared with bulk (0.024 µg/ml).

Silver Nanoparticles Stabilised by Cationic Gemini Surfactants with Variable Spacer Length.

The present study is focused on the synthesis and investigation of the physicochemical and biological properties of silver nanoparticles stabilized with a series of cationic gemini surfactants having a polymethylene spacer of variable length. UV-VIS spectroscopy, dynamic light scattering, scanning electron microscopy and zeta potential measurements were applied to provide physicochemical characterization of the silver nanoparticles. The mean size values of the nanoparticles were found to be in the 50 to 115 nm range. From the nanoparticle size distributions and scanning electron microscopy images it results that a population of small nanoparticles with the size of several nanometers was confirmed if the nanoparticles were stabilized with gemini molecules with either a short methylene spacer (two or four -CH2- groups) or a long spacer (12 -CH2- groups). The average zeta potential value for silver nanoparticles stabilized with gemini molecules is roughly independent of gemini surfactant spacer length and is approx. +58 mV. An interaction model between silver nanoparticles and gemini molecules which reflects the gained experimental data, is suggested. Microbicidal activity determinations revealed that the silver nanoparticles stabilized with gemini surfactants are more efficient against Gram-negative bacteria and yeasts, which has a direct relation to the interaction mechanism of nanoparticles with the bacterial cell membrane and its structural composition.

Preparation of Hydrochlorothiazide Nanoparticles for Solubility Enhancement.

Nanoparticles can be considered as a useful tool for improving properties of poorly soluble active ingredients. Hydrochlorothiazide (Class IV of the Biopharmaceutical Classification System) was chosen as a model compound. Antisolvent precipitation-solvent evaporation and emulsion solvent evaporation methods were used for preparation of 18 samples containing hydrochlorothiazide nanoparticles. Water solutions of surfactants sodium dodecyl sulfate, Tween 80 and carboxymethyl dextran were used in mass concentrations of 1%, 3% and 5%. Acetone and dichloromethane were used as solvents of the model compound. The particle size of the prepared samples was measured by dynamic light scattering. The selected sample of hydrochlorothiazide nanoparticles stabilized with carboxymethyl dextran sodium salt with particle size 2.6 nm was characterized additionally by Fourier transform mid-infrared spectroscopy and scanning electron microscopy. It was found that the solubility of this sample was 6.5-fold higher than that of bulk hydrochlorothiazide.

Preparation of risedronate nanoparticles by solvent evaporation technique.

One approach for the enhancement of oral drug bioavailability is the technique of nanoparticle preparation. Risedronate sodium (Biopharmaceutical Classification System Class III) was chosen as a model compound with high water solubility and low intestinal permeability. Eighteen samples of risedronate sodium were prepared by the solvent evaporation technique with sodium dodecyl sulfate, polysorbate, macrogol, sodium carboxymethyl cellulose and sodium carboxymethyl dextran as nanoparticle stabilizers applied in three concentrations. The prepared samples were characterized by dynamic light scattering and scanning electron microscopy. Fourier transform mid-infrared spectroscopy was used for verification of the composition of the samples. The particle size of sixteen samples was less than 200 nm. Polysorbate, sodium carboxymethyl dextran and macrogol were determined as the most favourable excipients; the particle size of the samples of risedronate with these excipients ranged from 2.8 to 10.5 nm.

Surface-active properties of nitrogen heterocyclic and dialkylamino derivates of hexadecylphosphocholine and cetyltrimethylammonium bromide.

The physico-chemical properties of dialkylamino and nitrogen heterocyclic analogues of hexadecylphosphocholine (HPC) and cetyltrimethylammonium bromide (CTAB) were investigated. The surface properties, such as the critical micelle concentration (cmc), the surface tension value at the cmc (gamma(cmc)), and the surface area at the surface saturation per head group (A(cmc)) were determined by means of surface tension measurements. Micelle size was determined using the dynamic light scattering method. The influence of dialkylamino groups and heterocyclic ring size on surface-active properties was investigated. Surface activity and micellar size of prepared analogues of HPC and CTAB were mutually compared.

Hydrodynamic size of DNA/cationic gemini surfactant complex as a function of surfactant structure.

The present study deals with the determination of hydrodynamic size of DNA/cationic gemini surfactant complex in sodium bromide solution using the dynamic light scattering method. Cationic gemini surfactants with polymethylene spacer of variable length were used for the interaction with DNA. The scattering experiments were performed at constant DNA and sodium bromide concentrations and variable surfactant concentration in the premicellar and micellar regions as a function of surfactant spacer length. It was found that the DNA conformation strongly depends on the polymethylene spacer length as well as on the surfactant concentration relative to the surfactant critical micelle concentration. Gemini surfactant molecules with 4 methylene groups in the spacer were found to be the least efficient DNA compacting agent in the region above the surfactant cmc. Gemini molecules with the shortest spacer length (2 methylene groups) and the longest spacer length (8 methylene groups) investigated showed the most efficient DNA compaction ability.

Biodegradable gemini surfactants. Correlation of area per surfactant molecule with surfactant structure.

Values of the area per surfactant molecule of various single chain and gemini quaternary ammonium surfactants containing biodegradable amide and ester groups are obtained from the surface tension measurements and they are mutually compared. It was found that surfactant molecules with the ester group in their structure occupy smaller area at the air/water interface than the corresponding molecules with the amide group, mainly due to the higher conformational flexibility of ester groups. In decreasing the area per surfactant molecule value, hydrogen bonding (both inter- and intramolecular) plays a significant role when amide groups are present in the spacer of a gemini molecule. They must be separated by a polymethylene chain or a flexible group such as cyclohexane which is short enough to allow intramolecular hydrogen bonds. The flexible cyclohexane group with the amide group in single chain surfactants may lead to the formation of intermolecular hydrogen bonds among surfactant molecules which also results in the reduction of the area per surfactant molecule.

Dynamic light scattering, interfacial properties, and conformational analysis of biodegradable quarternary ammonium surfactants.

Aggregation properties of biodegradable ammonium surfactants containing amide and ester groups in the bulk and at the air-water interface were investigated as a function of surfactant tail length m using dynamic light scattering and surface tension experimental methods. The results indicate that surfactants containing an ester group in the structure display higher aggregation ability in the volume and form more densely packed layer of molecules at the air-water interface than those with an amide group. The results of physical measurements were correlated with 3D models of respective surfactant molecules. As the results indicate, a surfactant molecule headgroup containing an ester group shows higher flexibility than that with an amide group in its structure, which is documented by somewhat smaller headgroup size and denser packing at the air-water interface.

Area per surfactant molecule values of gemini surfactants at the liquid-hydrophobic solid interface.

Areas per surfactant molecule at the liquid/hydrophobic solid (A(LS)) and the liquid/air (A(LA)) interface as a function of the spacer length are reported for cationic gemini surfactants having (CH2)n spacer s. A(LA) increases with increasing spacer length up to 6-8 CH2 groups in the spacer and then levels off. A(LS) values indicate a more closely packed arrangement of the surfactant molecules than that at the liquid/air interface. Comparison of A(LA) and A(LS) values indicates that the surfactant molecules at the liquid/hydrophobic solid interface are almost three times as closely packed as those at the liquid/air interface. A comparison of the experimental values of the area per surfactant molecule at both interfaces was made with those calculated from dimensions of the surfactant molecule in vacuo.

Interaction of gemini surfactants butane-1,4-diyl-bis(alkyldimethylammonium bromide) with DNA.

The size and structure of aggregates formed by interaction of DNA with homologous series of cationic gemini surfactants butane-1,4-diyl-bis(alkyldimethylammonium bromide) (CnGS, n=10-16 is the number of alkyl carbons) were investigated using UV-vis turbidity, dynamic light scattering and small-angle synchrotron X-ray (SAX) diffraction. The detailed analysis of turbidity in the range of lambda=450-600 nm indicates an anomaly in the growth of CnGS+DNA aggregates with increasing concentration of CnGS, possibly involving changes of structure and size of aggregates. Using dynamic light scattering, changes of the effective diameter of CnGS+DNA (n=12 and 16) aggregates formed in the CnGS concentration range 0.002-0.140 mmol/l were observed. SAX diffractograms show the presence of long-range organization of CnGS+DNA (n=12, 13, 14 and 16) aggregates due to DNA interaction with CnGS above the critical micellar concentration. The CnGS+DNA (n=12, 13 and 14) aggregates at 25 degrees C are packed in a lattice of two-dimensional hexagonal symmetry. With increasing C14GS:DNA molar ratio the changes of the lattice parameter in the range of 4.80-5.27 nm are observed at 25 degrees C. The aggregates undergo structural changes induced by temperature in the range 60-95 degrees C, which are accompanied by changes of the diffraction patterns, namely in the region of reciprocal spacing s=0.15-0.30 nm(-1).