Antileishmanial activity of amides from Piper amalago and synthetic analogs

Two natural amides isolated from the chloroform extract of Piper amalago L., Piperaceae, leaves, a hydrogenated derivative and seven synthetic analogs were tested against the promastigote and intracellular amastigote forms of Leishmania amazonensis. The antileishmanial activity was evaluated in terms of growth inhibitory concentration for 50% of protozoa (IC50). The cytotoxicity toward the J774A1 macrophages was evaluated in terms of the cytotoxic concentrations for 50% of macrophages (CC50). The ability to induce nitric oxide production was also investigated for all compounds. The saturated amide 7-(1,3-benzodioxol-5-yl)-1-(1-pyrrolidinyl)-1-heptanone was obtained by hydrogenation of the natural compound N-[7-(3',4'-methylenedioxyphenyl)-2(Z),4(Z)-heptadienoyl]pyrrolidine. Synthetic amides were prepared by addition of the appropriate amine to the corresponding acyl chloride. The natural compound, N-[7-(3',4'-methylenedioxyphenyl)-2(E),4(E)-heptadienoyl]pyrrolidine, was the most active of all tested compounds against the promastigote and intracellular amastigote forms with IC50 values of 15 µM and 14.5 µM, respectively. None of the compounds modulated the production of nitric oxide.

HPLC analysis of supercritical carbon dioxide and compressed propane extracts from Piper amalago L. with antileishmanial activity.

Piper amalago L. leaves were extracted with supercritical carbon dioxide and compressed propane under different conditions, and with chloroform by the conventional maceration method. These methods were compared for the pyrrolidine alkaloid content. Supercritical carbon dioxide (SFE-CO2) at 313 K and 12.55 MPa showed the highest selectivity for the main compound (600.53 mg/g of extract). A gradient high-performance liquid chromatography (HPLC) method was developed and validated to quantify the alkaloid N-[7-(3',4'-methylenedioxyphenyl)-2(Z),4(Z)-heptadienoyl]pyrrolidine in the extracts. The HPLC method showed linearity, precision and accuracy, allowing the quantitative analysis of the alkaloid in all the samples. All the extracts were tested against the promastigote and intracellular amastigote forms of Leishmania amazonensis. The antileishmanial activity was evaluated in terms of inhibitory concentration for 50% of protozoa (IC50). The cytotoxicity was also evaluated against J774A1 macrophages, and the cytotoxic concentrations for 50% of macrophages were obtained (CC50). The SFE-CO2 (313 K; 12.55 MPa) extract showed the highest antileishmanial activity with the following IC50 values of 16 and 7 µg/mL against the promastigotes and intracellular amastigotes forms, respectively. The extract showed low cytotoxicity with a CC50 value of 93 µg/mL.

Evaluation of soybean methanol fraction on acute inflammation.

Soybeans have been of interest of researchers because of the presence of isoflavones, a subclass of flavonoids, which have demonstrated anti-inflammatory activity. The aim of this study was investigate the anti-inflammatory activity of the methanol fraction from soybean, which contains mainly isoflavone glucosides and malonylglucosides. The anti-inflammatory activity of the methanol fraction from soybean was studied using croton oil-induced mouse ear edema and carrageenan-induced pleurisy models. The methanol fraction inhibited the ear edema in a dose-dependent manner: 0.625 mg/kg by 44.23% (P<.05), 1.25 mg/kg by 60.68% (P<.01), and 2.5 mg/kg by 65.68% (P<.01). Myeloperoxidase enzyme activity was reduced at the dose of 2.5 mg/kg (64.79%, P<.05). No effects were seen on carrageenan-induced pleurisy at different doses of the methanol fraction (100 or 400 mg/kg). These results demonstrated that the methanol fraction containing conjugated isoflavones showed topical anti-inflammatory activity. There was no acute toxicity in Swiss mice after oral administration of the fraction, at doses of 1,000, 2,000, 3,000, and 4,000 mg/kg.