Investigation of local anesthetic and antimycobacterial activity of Ottonia martiana Miq. (Piperaceae).

Ottonia martiana is a plant popularly known in Brazil by the use for toothache. Ethanolic extract (EE), hexane fraction (HF), dichloromethane fraction (DF) and piperovatine obtained from O. martiana were assayed in vitro and in vivo. The acute toxicity of EE was determined, and LD50 values of 164.5 and 65.0 mg/kg by the oral and intraperitoneal routes, respectively, indicated a high toxicity for EE in vivo, explaining its popular use by topical administration only. A local anesthetic-like effect of EE and its fractions was observed in experimental models using pain induction, and such effect involved an analgesic action. The antimycobacterial activity of EE, HF, DF and piperovatine was evaluated against Mycobacterium tuberculosis H37Rv ATCC 27924. EE, HF, DF, and piperovatine showed a potential antimycobacterial effect with MICs of 16.0, 62.0, 62.0 and 8.0 µg/mL, respectively. Piperovatine was more effective than the EE or the other fractions. The selectivity index (SI=IC50/MIC) values calculated for EE, HF, DF and piperovatine based on the MICs and the cytotoxicity against J774 macrophages (IC50 by MTT assay) revealed values of 6.43, 2.34, 1.5 and 9.66, respectively.

The natural compounds piperovatine and piperlonguminine induce autophagic cell death on Trypanosoma cruzi.

The currently available treatments for Chagas disease show limited therapeutic potential and are associated with serious side effects. Our group has been attempting to find alternative drugs isolated from natural products as a potential source of pharmacological agents against Trypanosoma cruzi. Here, we demonstrate the antitrypanosomal activity of the amides piperovatine and piperlonguminine isolated from Piper ovatum against epimastigotes and intracellular amastigotes. We also investigated the mechanisms of action of these compounds on extracellular amastigote and epimastigote forms of T. cruzi. These amides showed low toxicity to LLCMK(2) mammalian cells. By using transmission and scanning electron microscopy, we observed that the compounds caused severe alterations in T. cruzi. These alterations were mainly located in plasma membrane and mitochondria. Furthermore, the study of treated parasites labeled with Rh123, PI and MDC corroborate with our TEM data. These mitochondrial dysfunctions induced by the amides might trigger biochemical alterations that lead to cell death. Altogether, our data evidence a possible autophagic process.

1,3-diene probes for detection of triplet carbonyls in biological systems.

Electronically excited triplet carbonyls are formed during the oxidative degradation of polyunsaturated fatty acids, amino acids, and beta-dicarbonyl metabolites. Due to their long lifetime and high alkoxyl radical-like reactivity, triplet carbonyls may initiate deleterious reactions in biological systems. Here we study the quenching properties of conjugated dienes, specifically 2,4-hexadienoate (sorbate) and its alkyl ester, on triplet acetone generated chemically (thermolysis of tetramethyl-1,2-dioxetane) or enzymatically (horseradish peroxidase-catalyzed aerobic oxidation of isobutanal). Triplet acetone quenching rates were near diffusion control ( k q = 10 (8)-10 (9) M (-1) s (-1)) and accompanied by diene cis-trans isomerization. None of the dienes displays antioxidant activity in classical systems known to generate reactive oxygen species: superoxide anion radical, hydroxyl radical, alkoxyl and alkylperoxyl radicals, or singlet oxygen. Experiments with model systems used widely to study lipid peroxidation showed that sorbate can inhibit mitochondrial swelling induced by enzymically formed triplet benzophenone and quench the chemiluminescence of microsome preparations challenged with iron and ascorbate. Altogether, our data indicate that conjugated dienes can be used as specific quenchers of triplet carbonyls formed in biological systems during oxidative stress. Moreover, they suggest that the well-known food preservative properties of sorbate may be due to its triplet carbonyl quenching activity.