Raquel V. Vico

Multivalent Interaction of Cyclodextrin Vesicles, Carbohydrate Guests, and Lectins: A Kinetic Investigation†

An artificial glycocalix self-assembles when unilamellar bilayer vesicles of amphiphilic β-cyclodextrins are decorated with maltose- and lactose-adamantane conjugates by host-guest interactions. The maltose-decorated vesicles aggregate in the presence of lectin concanavalin A whereas the lactose-decorated vesicles aggregate in the presence of lectin peanut agglutinin. The kinetics of the orthogonal multivalent interfacial interactions present in this ternary system of vesicles, carbohydrates, and lectins were studied by time-dependent measurements of the optical density at 400 nm. The average vesicle and vesicle aggregate sizes were monitored by dynamic light scattering. The aggregation process was evaluated as a function of lectin concentration, vesicle concentration, and surface coverage of the vesicles by the carbohydrate-adamantane conjugates. The initial rate of vesicle aggregation scales linearly with the lectin as well as the cyclodextrin vesicle concentration. Furthermore, each lectin requires a characteristic critical density of carbohydrates at the vesicle surface. These observations allow a prediction of the response of the ternary supramolecular system at different concentrations of its components. Also, the effective binding site separation in a multivalent receptor such as a multiple binding site protein can be accurately determined. This methodology can be extended to multivalent noncovalent interactions in other ligand-receptor systems at interfaces.

Reactivity of the insecticide fenitrothion toward O and N nucleophiles.

The reactivity of Fenitrothion (1) toward several O- and N-based nucleophiles, including ambident and alpha-nucleophiles, was investigated in basic media at 25 degrees C in water containing 2% 1,4-dioxane. In the reactions with HO(-) and HOO(-) quantitative formation of 3-methyl-4-nitrophenoxide (2) was observed indicating a S(N)2(P) pathway. In the reactions with NH(2)OH, NH(2)O(-), and BuNH(2), demethylfenitrothion (4) was formed along with 2, indicating competition between the S(N)2(P) and S(N)2(C) pathways; no evidence of a S(N)Ar pathway was observed in any case. The observed rate constants were dissected into the values corresponding to the S(N)2(P) and S(N)2(C) pathways. The yield of 4 depends on the nucleophile and on the pH of the reaction, being the main product in the case of BuNH(2). With HOO(-), NH(2)OH, and NH(2)O(-) a significant alpha-effect was observed, confirming the participation of the nucleophile in the rate-limiting step of the reaction.

Distinctive Interactions of Oleic Acid Covered Magnetic Nanoparticles with Saturated and Unsaturated Phospholipids in Langmuir Monolayers

The growing number of innovations in nanomedicine and nanobiotechnology are posing new challenges in understanding the full spectrum of interactions between nanomateriales and biomolecules at nano-biointerfaces. Although considerable achievements have been accomplished by in vivo applications, many issues regarding the molecular nature of these interactions are far from being well-understood. In this work, we evaluate the interaction of hydrophobic magnetic nanoparticles (MNP) covered with a single layer of oleic acid with saturated and unsaturated phospholipids found in biomembranes through the use of Langmuir monolayers. We find distinctive interactions among the MNP with saturated and unsaturated phospholipids that are reflected by both, the compression isotherms and the surface topography of the films. The interaction between MNP and saturated lipids causes a noticeable reduction of the mean molecular area in the interfacial plane, while the interaction with unsaturated lipids promotes area expansion compared to the ideally mixed films. Moreover, when liquid expanded and liquid condensed phases of the phospholipid(s) coexist, the MNP preferably partition to the liquid-expanded phase, thus hindering the coalescence of the condensed domains with increasing surface pressure. In consequence organizational information on long-range order is attained. These results evidence the existence of a sensitive composition-dependent surface regulation given by phospholipid-nanoparticle interactions which enhance the biophysical relevance of understanding nanoparticle surface functionalization in relation to its interactions in biointerfaces constituted by defined types of biomolecules.

Alkyl esters of L-ascorbic acid: Stability, surface behaviour and interaction with phospholipid monolayers.

L-ascorbic acid alkyl esters (ASCn) are molecules of pharmaceutical interest for their amphiphilic nature and proposed antioxidant power. In contrast to L-ascorbic acid, ASC(n) with different acyl chain lengths behaved stably upon oxidation and a tautomeric isomerization was observed. In Langmuir films, when the ascorbic ring has a negative charge, ASC14 and ASC16 form stable monolayers, contrary to ASC10 and ASC12. ASC16 films showed transition from liquid-expanded (LE) to liquid-condensed phase, whereas ASC14 showed only an LE phase. When ASCn are mainly neutral, ASC14 showed phase transition from LE to a crystalline phase, as previously reported for ASC16. The two-dimensional domains displayed crystal-like shapes with anisotropic optical activity when interacting with the polarized light under Brewster angle microscopy. The compounds with the longer acyl chain (ASC16, ASC14 and ASC12) exhibited good surface activity, forming Gibbs monolayers. They also were able to penetrate into phospholipid monolayers up to a critical point of 45-50 mN/m. The 1-palmitoyl-2-oleoylphosphatidylcholine/ASCn films showed LC and/or crystalline domains only for ASC16. This study provides valuable evidence regarding the stability and surface properties of this drug family, and casts light into the differential interaction of these drugs with lipid membranes, which is important for understanding its differential pharmacological activity.

Molecular organization and recognition properties of amphiphilic cyclodextrins

The continuing challenge of using cyclodextrins (CDs) for solubilization and drug targeting has led to the preparation of a wide variety of chemically modified derivatives in order to improve the properties of these host molecules. A possible approach for pharmaceutical applications would be to combine the recognition specificity of CDs with the transport properties of organized structures such as vesicles, liposomes, or micelles. Amphiphilic CDs can be admixed to phospholipid monolayers and to liposomes, and they can be dispersed into nanospheres showing promising properties for drug encapsulation. Monoacylated derivatives of β-CD, Mod-CD (Cn), were synthesized in our laboratory from the reaction of alkenyl succinic anhydride with β-CD. We found that the compound with 10 carbon atoms in the alkenyl chain, Mod-CD (C10), can be incorporated into inverted micelles. We studied their properties in solution and at the air-water interface. In solution they have very low critical micellar concentration, and in the aggregates there are two recognition sites: one is the cavity of the CD and the other is formed by the hydrophobic tails. The alkenyl chain interacts with the cavity, but this is not an obstacle for the association with external guests such as 1-amino adamantane, phenolphthalein, or Prodan. Mod-CD (Cn) with n equal to 10, 14, and 16 (n indicates the number of carbons in the alkenyl chain), form stable monolayers at the air-water interface and they adopt an organization very different from those found for persubstituted CDs. The differences are attributed to the higher conformational flexibility of these compounds, which allows the organization of the CD units with the cavity perpendicular to the interface.

Effect of cyclodextrins on the reactivity of fenitrothion

The hydrolysis reaction of fenitrothion was studied in water containing 2% dioxane and in the presence of native cyclodextrins (α-, β- and γ-CD) and two commercially available modified derivatives, namely, permethylated β- and α-cyclodextrin (TRIMEB and TRIMEA, respectively). The kinetics of the reaction in the presence of TRIMEA could not be measured because the complex formed is insoluble and precipitated even at low concentration. On the other hand, the reaction is only weakly affected by the presence of α-CD. The hydrolysis reaction is inhibited by all the other cyclodextrins. From the kinetic data the association equilibrium constants for the formation of the 1:1 inclusion complexes were determined as 417, 511 and 99M(-1) for β-CD, TRIMEB and γ-CD, respectively. Despite the differences in the association constants for β- and γ-CD, the observed inhibition effect is about the same and this is due to the fact that the rate of hydrolysis in the cavity of γ-CD is smaller than that in the cavity of β-CD. The strongest inhibitor is TRIMEB and this result is consistent with the known structure of the complex in the solid state.

Solid-state structures and thermal properties of inclusion complexes of the organophosphate insecticide fenitrothion with permethylated cyclodextrins

The X-ray crystal structures and thermal stabilities of the inclusion complexes formed between the organophosphate insecticide fenitrothion [O,O-dimethyl O-(3-methyl-4-nitrophenyl) phosphorothioate] and the host compounds TRIMEA and TRIMEB (permethylated alpha- and beta-cyclodextrins, respectively) are reported. In the complex (TRIMEA)(2).fenitrothion 1, the guest phosphate ester group is disordered and the molecule is fully encapsulated within a novel TRIMEA dimer in which the secondary rims of the two host molecules are in close contact. In contrast, the complex TRIMEB.fenitrothion 2 is monomeric and the guest molecule is statistically disordered over two positions, with the phosphate group inserted in the host cavity in both cases. Thermal analysis indicated gradual and partial loss of the guest in 1 during heating between 130 degrees C and the melting point of the complex (approximately 200 degrees C), whereas complex 2 displayed significant mass loss only after fusion of the complex at 161 degrees C.

The Rheological Properties of Lipid Monolayers Modulate the Incorporation of l-Ascorbic Acid Alkyl Esters

In this work, we tested the hypothesis that the incorporation of amphiphilic drugs into lipid membranes may be regulated by their rheological properties. For this purpose, two members of the l-ascorbic acid alkyl esters family (ASCn) were selected, ASC16 and ASC14, which have different rheological properties when organized at the air/water interface. They are lipophilic forms of vitamin C used in topical pharmacological preparations. The effect of the phase state of the host lipid membranes on ASCn incorporation was explored using Langmuir monolayers. Films of pure lipids with known phase states have been selected, showing liquid-expanded, liquid-condensed, and solid phases as well as pure cholesterol films in liquid-ordered state. We also tested ternary and quaternary mixed films that mimic the properties of cholesterol containing membranes and of the stratum corneum. The compressibility and shear properties of those monolayers were assessed in order to define its phase character. We found that the length of the acyl chain of the ASCn compounds induces differential changes in the rheological properties of the host membrane and subtly regulates the kinetics and extent of the penetration process. The capacity for ASCn uptake was found to depend on the phase state of the host film. The increase in surface pressure resultant after amphiphile incorporation appears to be a function of the capacity of the host membrane to incorporate such amphiphile as well as the rheological response of the film. Hence, monolayers that show a solid phase state responded with a larger surface pressure increase to the incorporation of a comparable amount of amphiphile than liquid-expanded ones. The cholesterol-containing films, including the mixture that mimics stratum corneum, allowed a very scarce ASCn uptake independently of the membrane diffusional properties. This suggests an important contribution of Cho on the maintenance of the barrier function of stratum corneum.

PM-IRRAS Assessment of the Compression-Mediated Orientation of the Nanocavity of a Monoacylated β-Cyclodextrin in Monolayers at the Air−Water Interface

The structural orientation adopted along the compression-decompression isotherm by a monoacylated beta-cyclodextrin (C16-betaCD) at the air-water interface was assessed by polarization-modulation infrared reflection-adsorption spectroscopy (PM-IRRAS). The adoption of different orientations of the cyclic oligosaccharide unit, relative to the interfacial plane, was interpreted analyzing the PM-IRRAS band intensity ratios of specific vibrations corresponding to the cyclodextrin moiety as a function of the surface pressure for successive compression/decompression cycles. The spectroscopic analysis revealed that the cyclic oligosaccharide modifies its position under compression from one in which the plane of the cavity of the monoacylated beta-cyclodextrin lies almost parallel to the interface to another in which the plane of the cavity is perpendicular to the interface. Through the PM-IRRAS analysis, it was also possible to evidence the establishment of an intermolecular hydrogen bonding network that may play an important role in the dynamic properties of the monolayer packing. The hydrogen bonding network becomes more important with the increases of surface pressure, up to a molecular packing limit, and it imparts the surface properties of the film for future compression-decompression cycles.

Molecular Organization, Structural Orientation, and Surface Topography of Monoacylated β-Cyclodextrins in Monolayers at the Air−Aqueous Interface

The surface behavior of monoacylated beta-cyclodextrins, with hydrocarbon chains of 16, 14, and 10 carbons, has been assessed by the measurement of the surface pressure, surface (dipole) potential, optical reflectivity, and surface topography in monolayers at the air-water interface. For all the derivatives studied, the intermolecular organization adopted along compression-decompression isotherms reveals a rich variety of packing states which imply profound reorganization of the hydrophobic and hydrophilic moieties of the beta-cyclodextrin derivatives in the film, depending on the lateral surface pressure. The intermolecular arrangements are consistent with the adoption of a different and defined orientation of the cyclic oligosaccharide unit, relative to the interfacial plane and the aqueous subphase. This is different from the behavior of the per-substituted derivatives, and none of the changes exhibited by the monosubstituted forms are consistent with the oligosaccharide ring remaining in a fixed orientation along the interface when the surface pressure is varied.