Chemical profile and toxicogenetic safety assessment of Smallanthus sonchifolius (yacon) organic extracts.

The interest in Smallanthus sonchifolius (yacon) has strongly resurfaced due to its multiple beneficial effects on human health. This study aimed at determining the toxicity and the chemical profile of an ethanol extract (EE) and a crude lactone mixture (CLM) of yacon leaves. Cytotoxicity and genotoxicity tests were performed by the MTT assay and the alkaline version of the comet assay respectively. The phytochemical analysis, performed by chromatographic and spectroscopy techniques, revealed the presence of nine sesquiterpene lactones (STLs) and two acyclic diterpene acids. In all cases, cell viability was inversely proportional to the extract concentration employed. The effects obtained with the highest dose of EE were significantly different from those obtained with the negative and solvent controls. Conversely, no significant differences were observed between the lowest doses of EE and controls. As for CLM, all tested doses showed statistically significant increases, as compared to negative and solvent controls.

Toxicogenetic evaluation of Smallanthus sonchifolius (yacon) as a herbal medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Smallanthus sonchifolius (Poepp. & Endl.) H. Robinson, commonly known as yacon, is a medicinal plant belonging to the Asteraceae family used in traditional folk medicine. Its roots and leaves have been used by people suffering from diabetes or from various digestive or renal disorders. AIM OF THE STUDY: This study aimed at evaluating the in vitro potential genotoxic effects of the aqueous extract of yacon in order to determine its safety and at characterizing its phytochemical composition. MATERIALS AND METHODS: The aqueous extract of S. sonchifolius was prepared in a similar way to that commonly used in popular medicine as tea bags. Thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC-MS/MS) were used to identify the main compounds. The MTT test was performed to determine the range of doses and the Cytochalasine B-blocked micronucleus (Cytome assay) was used to assess geneotoxicity. RESULTS: The chemical analysis of the aqueous extract revealed the presence of the sesquiterpene lactones (STLs) enhydrin and the dimer enhydrofolin, as the main compounds together with phenolic compounds. Increasing concentrations of the extract induced a cytotoxic effect on CHO-K1 and HepG2 cells. A statistically significant increase in the frequency of MNi, NBUDs and NPBs was observed in CHO-K1 cells, while in HepG2 cells a statistically significant frequency increase was observed with three of the four tested doses for MNi and only with the highest dose for NPBs and NBUs (genotoxic effect). CONCLUSION: Results demonstrated the inability of the metabolic system to counteract the genetic instability, allowing the safe consumption of the leaves as a 2% tea infusion in quantities of up to 250 mL/day.

Adulterants in selected dietary supplements and their detection methods.

At present, there is a growing trend toward the intentional adulteration of dietary supplements (DS) with synthetic pharmaceuticals, which represents an alarming emerging risk to consumers and a serious problem for regulatory agencies. An amazing array of synthetic drugs and their analogues have been reported as adulterants in DS. Mainly, the presence of analogues represents a serious health risk as their efficacy and toxic effects have not been clinically assessed yet and may result in unpredictable adverse effects. The purpose of this review is to provide an overview, over the period 2009-2019, of the most frequently reported adulterants in DS for the treatment of erectile dysfunction, obesity/overweight, diabetes mellitus, and hypertension and the analytical methods used for their detection.

Germacranolide-type sesquiterpene lactones from Smallanthus sonchifolius with promising activity against Leishmania mexicana and Trypanosoma cruzi.

BACKGROUND: Leishmaniasis and Chagas disease are life-threatening illnesses caused by the protozoan parasites Leishmania spp. and Trypanosoma cruzi, respectively. They are known as "neglected diseases" due to the lack of effective drug treatments and the scarcity of research work devoted to them. Therefore, the development of novel and effective drugs is an important and urgent need. Natural products are an important source of bioactive molecules for the development of new drugs. In this study, we evaluated the activity of enhydrin, uvedalin and polymatin B, three sesquiterpene lactones (STLs) isolated from Smallanthus sonchifolius, on Leishmania mexicana (MNYC/BZ/62/M) and Trypanosoma cruzi (Dm28c). In addition, the in vivo trypanocidal activity of enhydrin and uvedalin and the effects of these STLs on parasites' ultrastructure were evaluated. METHODS: The inhibitory effect of the three STLs on the growth of L. mexicana amastigotes and promastigotes as well as T. cruzi epimastigotes was evaluated in vitro. The changes produced by the STLs on the ultrastructure of parasites were examined by transmission electron microscopy (TEM). Enhydrin and uvedalin were also studied in a murine model of acute T. cruzi infection (RA strain). Serum activities of the hepatic enzymes alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase were used as biochemical markers of hepatotoxicity. RESULTS: The three compounds exhibited leishmanicidal activity on both parasite forms with IC50 values of 0.42-0.54 µg/ml for promastigotes and 0.85-1.64 µg/ml for intracellular amastigotes. Similar results were observed on T. cruzi epimastigotes (IC50 0.35-0.60 µg/ml). The TEM evaluation showed marked ultrastructural alterations, such as an intense vacuolization and mitochondrial swelling in both L. mexicana promastigotes and T. cruzi epimastigotes exposed to the STLs. In the in vivo study, enhydrin and uvedalin displayed a significant decrease in circulating parasites (50-71%) and no signs of hepatotoxicity were detected. CONCLUSIONS: Enhydrin, uvedalin and polymatin B possess significant leishmanicidal and trypanocidal activity on different parasite stages. These results show that these compounds may provide valuable leads for the development of new drugs against these neglected parasitic diseases.

Natural Sesquiterpene Lactones as Potential Trypanocidal Therapeutic Agents: A Review.

Chagas' disease and Human African Trypanosomiasis are parasitic diseases that remain major health problems, mainly among the poorest and the most marginalized communities from Latin America and Africa. The scarcity of effective chemotherapy, due to the low investment in the research and development (R&D) of new drugs, together with a high incidence of side effects, and the emergence of drug resistance phenomena emphasize the urgent need for new prophylactic and therapeutic agents. Over the ages, humans have employed natural products to treat a wide spectrum of diseases. Recently, the pharmaceutical industry has focused on plant research and a large body of evidence has been collected to demonstrate the immense potential of medicinal plants as a source of bioactive compounds and lead molecules. In the field of parasitic diseases, drug development from plants has been successful for the sesquiterpene lactone (STL) artemisinin, which is employed as an antimalarial agent. STLs are a large group of naturally occurring terpenoids derived from plants that mostly belong to the Asteraceae family which exhibit a variety of skeletal arrangements and are the largest and most diverse category of natural products with an a- m6thylene-lactone motif. STLs display a broad spectrum of biological activities such as antitumor, cytotoxic, antibacterial, anthelmintic, uterus contracting, antimalarial, neurotoxic, antiprotozoal and allergic (contact dermatitis) activities. In this context, the purpose of the present review is to provide an overview of the trypanocidal activity reported for STLs against Trypanosoma cruzi and T. brucei rhodesiense over the period 1993-2015.

The sesquiterpene lactone polymatin B from Smallanthus sonchifolius induces different cell death mechanisms in three cancer cell lines.

A 8ß-angeloyloxy-9α-hydroxy-14-oxo-acanthospermolide and five known melampolide sesquiterpene lactones (uvedalin, enhydrin, polymatin B, sonchifolin, and fluctuanin) were isolated from the leaves of Smallanthus sonchifolius. The compounds were identified by 1D-, 2D-NMR, HRMS, IR and UV analyses. In vitro cytotoxicity assays (MTT) showed that these sesquiterpene lactones display poor cytotoxic effects on peripheral blood mononuclear cells (PBMC) of healthy human subjects, whereas a strong cytotoxicity was observed in leukemia and pancreas cancer cells. For the mechanism of action of polymatin B, oxidative stress seems to be involved. Interestingly, reactive oxygen species (ROS) formation mainly induced different effects: apoptosis in CCRF-CEM cells, necroptosis in CEM-ADR5000 cells through induction of RIP1K, neither apoptosis nor necroptosis in MIA-PaCa-2 cells. Additionally, cells also died partly by necrosis.

Detection of a tadalafil analogue as an adulterant in a dietary supplement for erectile dysfunction.

INTRODUCTION: Several cases of adulteration of dietary supplements with tadalafil, sildenafil, and vardenafil, or their unapproved analogues have been reported worldwide. Mainly, the presence of the latter represents a serious health risk to consumers as their efficacy and toxic effects have not been assessed and may result in unpredictable adverse effects. AIM: To investigate the suspected adulteration with synthetic phosphodiesterase type 5 (PDE-5) inhibitors in a dietary supplement marketed in Argentina for the treatment of erectile dysfunction (ED). METHODS: The content of the capsules of the dietary supplement (sample A) was analyzed by thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) diode-array detection. From the organic extract of sample A, a major compound was purified by column chromatography (CC). The isolated compound was identified by proton nuclear magnetic resonance (1H NMR) and carbon NMR (13C NMR), heteronuclear single quantum coherence, distortionless enhancement by polarization transfer (DEPT 135), electrospray ionization mass spectrometry, and ultraviolet, and infrared (Fourier transform infrared spectroscopy) spectroscopy. MAIN OUTCOME MEASURE: Proof of adulteration of herbal products with synthetic PDE-5 inhibitors. RESULTS: By TLC and HPLC analysis, a major compound was detected in sample A organic extract. The purification of this extract by CC led to the isolation of a pure compound which was identified according to its spectral data as (6R,12aR)-2-amino-6-(1,3-benzodioxol-5-yl)-2,3,6,7,12,12a-hexahydropyrazino [1',2':1,6] pyrido [3,4-b] indole-1,4-dione or aminotadalafil. CONCLUSIONS: An unapproved PDE-5 inhibitor analogue, which was identified as aminotadalafil, has been detected in a dietary supplement. This study represents the first report in Latin America and one of the few independent studies of an adulteration with an unapproved PDE-5 inhibitor of an herbal product for ED treatment.