Conjugation of triphenylantimony(V) with carvacrol against human breast cancer cells.

The organoantimony derivative of formula trans-O,O-[Ph3SbV(Carv)2] (TPAC) (CarvH = carvacrol) is obtained by the oxidation of triphenylstibine (Ph3SbIII) with hydrogen peroxide in the presence of carvacrol (CarvH). Physical methods such as X-ray Fluorescence (XRF) spectroscopy, single crystal and powder X-ray diffraction analysis (XRD and PXRD), Attenuated Total Reflection Fourier Transform Infra-red (ATR-FTIR) spectroscopy, Thermogravimetric Differential Thermal Analysis (TG-DTA) and Differential Scanning Calorimetry (DTG/DSC), confirm the retention of the formula of TPAC throughout the sample mass in solid state, while UV-Vis spectroscopy in the solution. TPAC is the first example of carvacrol (the main ingredient of oregano) covalently bonded to any metal ion. Only the trans-O,O-[Ph3Sb(Carv)2] isomer was isolated suggesting stereo-selectivity of the preparation route. TPAC inhibits in vitro both human breast adenocarcinoma cell lines: MCF-7 (positive to hormones receptor (HR +)), MDA-MB-231 (negative to hormones receptor (HR-)) stronger than normal human fetal lung fibroblast cells (MRC-5). The MCF-7 cells morphology, DNA fragmentation, Acridine Orange/Ethidium Bromide (AO/EB) Staining, cell cycle arrest and mitochondrial membrane permeabilization tests suggest an apoptotic pathway for cell death, especially, through the mitochondrion damage. The binding type of TPAC toward the calf thymus CT-DNA was initially deduced ex vivo from the differentiation of the DNA solution viscosity. Fluorescence spectroscopy confirms the interaction mode suggested. Spectroscopic evidence (FTIR, UV-Vis) suggest that glutathione (GSH) (a tripeptide over-expressed in tumor cells) induces conversion of non-active pentavalent antimony, which is contained in TPAC, to active trivalent one, providing a new strategy for the development of targeted chemotherapeutics.

Immobilization of Lipid Substrates: Application on Phospholipase A2 Determination.

The purpose of the study was to assess a fluorimetric assay for the determination of total phospholipase A(2) (PLA(2)) activity in biological samples introducing the innovation of immobilized substrates on crosslinked polymeric membranes. The immobilized C(12)-NBD-PtdCho, a fluorescent analogue of phosphatidylcholine, exhibited excellent stability for 3 months at 4 °C and was not desorbed in the aqueous reaction mixture during analysis. The limit of detection was 0.5 pmol FA (0.2 pg) and the linear part of the response curve extended from 1 up to 190 nmol FA/h/mL sample. The intra- and inter-day relative standard deviations (%RSD), were ≤6 and ≤9 %, respectively. Statistical comparison with other fluorescent methods showed excellent correlation and agreement. Semiempirical calculations showed a fair amount of electrostatic interaction between the NBD-labeled substrate and the crosslinked polyvinyl alcohol with the styryl pyridinium residues (PVA-SbQ) material, from the plane of which, the sn-2 acyl chain of the phospholipid stands out and is accessible by PLA(2). Atomic Force Microscopy revealed morphological alterations of the immobilized substrate after the reaction with PLA(2). Mass spectrometry showed that only C(12)-NBD-FA, the PLA(2 )hydrolysis product, was detected in the reaction mixture, indicating that PLA(2) recognizes PVA-SbQ/C(12)-NBD-PtdCho as a surface to perform catalysis.