No abstract

Valsartan is a marketed drug with high affinity to the type 1 angiotensin (AT1) receptor. It has been reported that AT1 antagonists may reach the receptor site by diffusion through the plasma membrane. For this reason we have applied a combination of differential scanning calorimetry (DSC), Raman spectroscopy and small and wide angle X-ray scattering (SAXS and WAXS) to investigate the interactions of valsartan with the model membrane of dipalmitoyl-phosphatidylcholine (DPPC). Hence, the thermal, dynamic and structural effects in bulk as well as local dynamic properties in the bilayers were studied with different valsartan concentrations ranging from 0 to 20 mol%. The DSC experimental results showed that valsartan causes a lowering and broadening of the phase transition. A splitting of the main transition is observed at high drug concentrations. In addition, valsartan causes an increase in enthalpy change of the main transition, which can be related to the induction of interdigitation of the lipid bilayers in the gel phase. Raman spectroscopy revealed distinct interactions between valsartan with the lipid interface localizing it in the polar head group region and in the upper part of the hydrophobic core. This localization of the drug molecule in the lipid bilayers supports the interdigitation view. SAXS measurements confirm a monotonous bilayer thinning in the fluid phase, associated with a steady increase of the root mean square fluctuation of the bilayers as the valsartan concentration is increased. At high drug concentrations these fluctuations are mainly governed by the electrostatic repulsion of neighboring membranes. Finally, valsartans' complex thermal and structural effects on DPPC bilayers are illustrated and discussed on a molecular level.

The structural modifications of the amino acid DL-Norvaline have been studied using differential scanning calorimetry (DSC) and Raman spectroscopy. DSC results showed that this amino acid undergoes two solid-solid phase transitions at -116.9 and -76.1 degrees C in the temperature range -130 to +300 degrees C. Raman spectroscopy was applied to complement DSC results. The combination of the two methodologies point out that the observed phase transitions correspond to an increment of disordering in the aliphatic side chain of amino acid, an augmentation of the rotational motion of the amino group and a decrease of the strength of the intramolecular hydrogen bonding of the initial dimers at low temperatures. The observed phase transitions of DL-norvaline are compared with those found in DL-norleucine.

Cyclodextrins (CDs) are a well-known class of supermolecules that have been widely used to protect drugs against conjugation and metabolic inactivation as well as to enhance the aqueous solubility and hence to ameliorate the oral bioavailability of sparingly soluble drug molecules. The hepatoprotectant drug silibinin can be incorporated into CDs, and here we elucidate the interaction between the drug and the host at the molecular level. The complexation product of silibinin with 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) is characterized by Differential Scanning Calorimetry, mass spectrometry, solid and liquid high-resolution NMR spectroscopy. The chemical shift changes using (13)C CP/MAS on the complexing of the guest with the host provided significant information on the molecular interactions, and they were in agreement with the 2D NOESY results. These results point out that in both solid and liquid forms, the drug is engulfed and interacts with HP-β-CD in identical manner. Molecular dynamics calculations have been performed to examine the thermodynamic characteristics associated with the silibinin-HP-β-CD interactions and to study the stability of the complex. To approximate the physiological conditions, the aqueous solubility and dissolution characteristics of the complex at pH states simulating those of the upper gastrointestinal tract have been applied. To evaluate the antiproliferative activity of silibinin-HP-β-CD complex comparatively to silibinin in MCF-7 human cancer cells, MTT assays have been performed.

A cross-polarization (CP) (31)P NMR broadline simulation methodology was developed for studying the effects of drugs in phospholipids bilayers. Based on seven-parameter fittings, this methodology provided information concerning the conformational changes and dynamics effects of losartan in the polar region of the dipalmitoylphosphatidylcholine bilayers. The test molecule for this study was losartan, an antihypertensive drug known to exert its effect on AT(1) transmembrane receptors. The results were complemented and compared with those of differential scanning calorimetry, solid-state (13)C NMR spectroscopy, Raman spectroscopy, and electron spin resonance. More specifically, these physical chemical methodologies indicated that the amphipathic losartan molecule interacts with the hydrophilic-head zone of the lipid bilayers. The CP (31)P NMR broadline simulations showed that the lipid molecules in the bilayers containing losartan displayed greater collective tilt compared to the tilt displayed by the load-free bilayers, indicating improved packing. The Raman results displayed a decrease in the trans/gauche ratio and increased intermolecular interactions of the acyl chains in the liquid crystalline phase. Additional evidence, suggesting that losartan possibly anchors in the realm of the headgroup, was derived from upfield shift of the average chemical shift sigma(iso) of the (31)P signal in the presence of losartan and from shift of the observed peak at 715 cm(-1) attributed to C-N stretching in the Raman spectra.

A methodology has been developed to detect partial interdigitation of lipid bilayers when a bioactive molecule is intercalated between the polar, interface, or hydrophobic segments. This methodology uses the easily accessible differential scanning calorimetry (DSC) technique as a screening one and the increase of DH due to the incorporated drug in lipid bilayers as a diagnostic thermodynamic parameter. The combined use of X-ray diffraction and Raman spectroscopy complement and confirm the provided by DSC information as it is shown in three classes of molecules, namely AT 1 antagonists, vinca alkaloids, and anesthetic steroids. For the two classes of molecules, AT 1 antagonists and vinca alkaloids, their presence in lipid bilayers results in the increase of DH and it is accompanied by the increase of trans:gauche ratio and the decrease of d-spacing as depicted by Raman spectroscopy and small-angle X-ray diffraction correspondingly, confirming the predictive ability of DSC experiments. When an anesthetic steroid is incorporated in lipid bilayers, neither increase of DH nor decrease of d-spacing was observed, confirming again the DSC results that show the absence of partial interdigitation of this class of molecules. Molecular dynamics simulations have been carried out for a representative system [(5S)-1-benzylo-5-(1H-benzimidazol-1-ylo-methylo)-2-pyrrolidinone (MMK3) ligands at 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipid bilayer], and the results confirmed the experimental findings. The change of distance at z-axis of oxygen atoms at head group of lipid molecule has been measured throughout the simulations. Statistical analysis has shown $8.8 Å interdigitation. Derived computational results are encouraging and can be performed to another ligand/lipid system. The development of a theoretical methodology will lead to advance the field and save a valuable time and effort.

In this work, syndiotactic polypropylene/multiwalled carbon nanotubes (MWCNT) nanocomposites, in various concentrations, were produced using melt mixing. The influence of the addition of MWCNT on the morphology, crystalline form, and the thermal and electrical properties of the polymer matrix was studied. To that aim, scanning electron microscopy, Raman spectroscopy, X-ray diffraction, differential scanning calorimetry, and dielectric relaxation spectroscopy were employed. Significant alterations of both the crystallization behavior and the thermal properties of the matrix were found on addition of the carbon nanotubes: conversion of the disordered crystalline form I to the ordered one, increase of the crystallization temperature and the degree of crystallinity, and decrease of the glass transition temperature and the heat capacity jump. Finally, the electrical percolation threshold was found between 2.5-3.0 wt.% MWCNT. For comparison purposes, the results of the system studied here are also correlated with the findings from a previous work on the isotactic polypropylene/MWCNT system.

It is proposed that AT1 antagonists (ARBs) exert their biological action by inserting into the lipid membrane and then diffuse to the active site of AT1 receptor. Thus, lipid bilayers are expected to be actively involved and play a critical role in drug action. For this reason, the thermal, dynamic and structural effects of olmesartan alone and together with cholesterol were studied using differential scanning calorimetry (DSC), 13C magic-angle spinning (MAS) nuclear magnetic resonance (NMR), cross-polarization (CP) MAS NMR, and Raman spectroscopy as well as small- and wide angle X-ray scattering (SAXS and WAXS) on dipalmitoyl-phosphatidylcholine (DPPC) multilamellar vesicles. 13C CP/MAS spectra provided direct evidence for the incorporation of olmesartan and cholesterol in lipid bilayers. Raman and X-ray data revealed how both molecules modify the bilayer's properties. Olmesartan locates itself at the head-group region and upper segment of the lipid bilayers as 13C CP/MAS spectra show that its presence causes significant chemical shift changes mainly in the A ring of the steroidal part of cholesterol. The influence of olmesartan on DPPC/cholesterol bilayers is less pronounced. Although, olmesartan and cholesterol are residing at the same region of the lipid bilayers, due to their different sizes, display distinct impacts on the bilayer's properties. Cholesterol broadens significantly the main transition, abolishes the pre-transition, and decreases the membrane fluidity above the main transition. Olmesartan is the only so far studied ARB that increases the gauche:trans ratio in the liquid crystalline phase. These significant differences of olmesartan may in part explain its distinct pharmacological profile.