After these experimental analyses, all lactones compounds were submitted to ab initio quantum calculations (DFT – Density Functional Theory – UB3LYP/6-31G*) and the values

of their physical–chemistry properties were analyzed by chemometric methods, in order to recognize patterns that correlate the lactone structures with their biological activities. The results obtained may aid in the development of new selective inhibitors for phospholipases A2 and, consequently, Selleck PD 332991 the treatment of poisoning by snake bites. All reagents, including Lac01 (α-santonin), were purchased from Aldrich or Sigma Co (USA). B. jararacussu venom was purchased from a private serpentarium in Formiga, MG, Brazil. B. jararacussu PLA2 was isolated employing two chromatographic steps: first gel filtration on Sephadex G-75, followed by cation-exchange chromatography. The column was previously equilibrated with 0.05 M ammonium bicarbonate buffer, pH 8.0. Elution was carried out with a continuous gradient up to a concentration

of 0.5 M ammonium bicarbonate. Absorbance of the effluent solution was recorded at a wavelength of 280 nm. PLA2 homogeneity was assessed by native and SDS-PAGE and reverse-phase selleckchem HPLC. Fraction II, known as Asp49 BthTX-II, was used in this study. This phospholipase will be denominated in this paper as just PLA2 ( Da Silva et al., 2008a and Da Silva et al., 2008b). Male Swiss mice, 6–8

weeks old, were matched for body weight (18–22 g). The animals were housed for at least one week before the experiment in laminar-flow cages maintained at a temperature of 22 ± 2 °C and a relative humidity of 50–60%, under a 12:12 h light–dark cycle. The animal experiments were carried out with the approval of the institutional committee of ethics, in accordance with protocols following the recommendations of the Canadian Council on Animal Care. The mice used in this study Cytidine deaminase were kept under specific pathogen-free conditions. The compounds employed in this study are shown in Fig. 1. Lactones 2, 3, 5, 6, 7, and 8 were prepared by procedures described in the literature (Arantes et al., 2009 and De Alvarenga et al., 2009). Lac04 was prepared as described below. To characterization of Lac04: IR spectra were recorded on a Perkin Elmer Paragon 1000 FTIR spectrophotometer, KBr, νmax, cm−1. 1H and 13C NMR spectra were obtained on a Bruker AVANCE DRX400 spectrometer at 400 and 100 MHz, respectively, and a Varian Mercury spectrometer observing 1H at 300 MHz and 13C at 75 MHz. All 1H and 13C spectra were obtained using CDCl3 as solvent and TMS as internal standard. Low resolution mass spectra were obtained on a SHIMADZU GC MS-QP5050A instrument by direct injection. The microanalysis was obtained on a PERKIN ELMER 2400 instrument.