Phytochemical Analysis, α-Glucosidase and α-Amylase Inhibitory Activities and Acute Toxicity Studies of Extracts from Pomegranate (Punica granatum) Bark, a Valuable Agro-Industrial By-Product.

Punica granatum is a tree of the Punicaceae family which is widespread all over the world with several types of varieties. Its fruit juice is highly prized, whereas the bark, rich in in phytochemicals such as flavonoids, hydrolysable tannins, phenolic acids, and fatty acids, is regarded an agro-industrial waste. It is utilized in traditional medicine for its medicinal properties in the treatment and prevention of a variety of ailments. This study aims to extract and to separate the phytochemical compounds from the bark of P. granatum, to identify them and to study the inhibitory effect of its extracts against antidiabetic activity. First, we carried out successive hot extractions with solvents (chloroform, acetone, methanol, and water) of increasing polarity by the Soxhlet. Then, using both qualitative and quantitative phytochemical investigation, we were able to identify groups of chemicals that were present in all extracts. We identified the majority of the molecular structures of chemicals found in each extract using HPLC-DAD analysis. The inhibition against both intestinal α-glucosidase and pancreatic α-amylase enzymes by P. granatum extracts was used to evaluate their potential antidiabetic effect in vitro. Our results demonstrated the great potential of the acetone extract. Ellagic acid, (-)-catechin, vanillin and vanillic acid were proposed as the most active compounds by the correlation analysis, and their actions were confirmed through the calculation of their IC50 and the determination of their inhibition mechanisms by molecular modelling. To summarize, these results showed that P. granatum bark, a natural agro-industrial by-product, may constitute a promising option for antidiabetic therapeutic therapy.

Phytochemical Profile, α-Glucosidase, and α-Amylase Inhibition Potential and Toxicity Evaluation of Extracts from Citrus aurantium (L) Peel, a Valuable By-Product from Northeastern Morocco.

Due to the high volume of peel produced, Citrus by-product processing could be a significant source of phenolic compounds, in addition to essential oil. Citrus fruit residues, which are usually dumped as waste in the environment, could be used as a source of nutraceuticals. Citrus aurantium (L), also known as sour or bitter orange, is a member of the Rutaceae family and is the result of interspecific hybridization between Citrus reticulata and Citrus maxima. The purpose of this study is to chemically and biologically evaluate the peel of C. aurantium, which is considered a solid waste destined for abandonment. To achieve more complete extraction of the phytochemicals, we used a sequential extraction process with Soxhlet using the increasing polarity of solvents (i.e., cyclohexane, chloroform, ethyl acetate, acetone, and ethanol-water mixture). Essential oil (EO) from the Citrus peel, which was present at 1.12%, was also prepared by hydrodistillation for comparison. Various phytochemical assays were used to determine the qualitative chemical composition, which was subsequently characterized using GC-MS and HPLC-DAD. The inhibitory effects of C. aurantium peel extract on two enzymes, intestinal α-glucosidase and pancreatic α-amylase, were measured in vitro to determine their potential hypoglycemic and antidiabetic actions. Each extract had a significantly different phytochemical composition. According to GC-MS analyses, which allow the identification of 19 compounds, d-limonene is the most abundant compound in both EO and cyclohexane extract, at 35.17% and 36.15% (w/w). This comparison with hydrodistillation shows the value of the sequential process in extracting this valuable terpene in large quantities while also allowing for the subsequent extraction of other bioactive substances. On the contrary, linoleic acid is abundant (54.35% (w/w)) in ethyl acetate extract (EAE) with a lower amount of d-limonene. HPLC-DAD analysis allows the identification of 11 phytochemicals, with naringenin being the most abundant flavanone, detected in acetone extract (ACE) (23.94% (w/w)), ethanol-water extract mixture (EWE) (28.71% (w/w)), and chloroform extract (CFE) (30.20% (w/w)). Several extracts significantly inhibited α-amylase and/or α-glycosidase in vitro. At a dose of 332 g/mL, ACE, CFE, and EWE inhibited the two enzymes by approximately 98%. There were strong significant correlations between naringenin and α-glucosidase inhibition and between gallic acid and α-amylase inhibition. Molecular docking experiments further verified this. Finally, oral administration of C. aurantium extracts at a dose of 2000 mg/kg did not cause any effect on mice mortality or signs of acute toxicity, indicating that it is non-toxic at these doses. These findings suggest that C. aurantium peels could be a valuable by-product by providing a rich source of non-toxic phytoconstituents, particularly those with potential antidiabetic action that needs to be confirmed in vivo.

New azole antifungal agents with novel modes of action: synthesis and biological studies of new tridentate ligands based on pyrazole and triazole.

The synthesis and extensive biological study of two new tridentates ligands based on pyrazole and triazole are described. The antifungal activity against the budding yeast cells of the newly synthesized compounds was determined. These compounds were toxic to yeast cells. Cell cycle analysis suggested that treatment with these compounds impairs cell division in G1 of the cell cycle. Using yeast-based functional genomics technologies, we found that these compounds tolerance requires DNA repair pathway and SKI complex function. We have also found that the PKC1 heterozygous deletion strain was the most sensitive to these compounds using HaploInsufficiency Profiling, suggesting that the Pkc1 protein may be the target for these compounds. These results strongly suggest that these compounds induce DNA damage and thus exert a different mechanism of action compared to other azole derivatives. These two compounds might therefore represent promising lead compounds for further development of antifungal drugs for human therapy.