Antioxidant and anti-inflammatory activity through inhibition of NF-κB and sEH of some citrus peel and phytoconstituent characteristics.

In Indonesia, there are many types of citrus where parts of the fruit, leaves, and peel can be utilized as food, drinks, spices, and medicine. This research aims to determine the phytochemical characteristics, antioxidant activities, and anti-inflammatory activity through inhibition of NF-κB and sEH, and the main phytoconstituents of three types of citrus fruits that are commonly used as herbs in Indonesia. The flesh and peel of Citrus amblycarpa/CAm, C. aurantiifolia/CAu, and C. hystrix/CH were extracted by Ultrasound-Assisted Extraction (UAE) with 70 % ethanol and then concentrated. All extracts were tested for total flavonoid content (TFC), total polyphenolic content (TPC), chemical constituents using LCMS, and DPPH radical scavenging activity. Molecular docking tests of 33 compounds containing CAm, CAu, and CH fruit peels from the literature study against NF-κB (Nuclear Factor Kappa Beta) and sEH (Soluble Epoxide Hydrolase) were also conducted. The TFC in fruit peels was 13.47-17.34 mg QE/g extract, and in flesh was 1.35-2.51 mg QE/g extract. The TPC in fruit peels was 4.28-6.3 mg GAE/g extract, and in flesh was 0.85-2.09 mg GAE/g extract. The IC50 values of antioxidant activity on fruit peel were 74.01-168.54 µg/mL; and flesh 185.62-2669 µg/mL. CAu peels provided the highest antioxidant activity and polyphenol content. The LC-MS/MS test on citrus peels shows the main chemical compounds: naringin (C27H32O14), naringenin (C15H12O5), hesperidin (C28H34O15), and hesperitin (C16H14O6). Molecular docking shows that naringin and neohesperidin predicted inhibit NF-κB, and hesperidin, neohesperidin, narirutin, naringin, apigenin, kaempferol, quercetin, rutin, eriocitrin, sinensetin, and vitamin A predicted can inhibit sEH enzyme. All citrus peel has stronger antioxidant activity and more flavonoids and phenolics than the flesh. Naringin and neohesperidin can inhibit NF-κB and sEH enzymes. The main flavonoid contents of the citrus peels and presumed to have activity are hesperidin and naringin. These flavonoids and their glycosides can be used as marker phytoconstituents in the quality assurance of pharmaceutical products.

Investigation of Yacon Leaves (Smallanthus sonchifolius) for α-Glucosidase Inhibitors Using Metabolomics and In Silico Approach.

Yacon (Smallanthus sonchifolius (Poepp.) H. Robinson) leaves is traditionally consumed as herbal tea in many countries including Indonesia. This plant's antidiabetic properties have been extensively researched, but studies on the responsible active compound identification are scarce. Information on the active compounds is critical for the consistency of Yacon herbal tea quality. The aim of this study was to identify α-glucosidase inhibitors in Indonesian Yacon leaves grown in two different locations using FTIR- and LC-MS/MS-based metabolomics in combination with in silico technique. Yacon leaves ethanol (50 and 95%) and water extracts were tested for α-glucosidase inhibitory activity, with the 95% ethanol extract being the most active. Geographical origins were found to have no major impact on the activity. In parallel, chemical profile of Yacon leaves extract was determined using FTIR and LC-MS/MS. Orthogonal Projection to Latent Structure (OPLS) was used to analyze both sets of data. OPLS analysis of FTIR data showed that compounds associated to α-glucosidase inhibitor activity included those with functional groups -OH, stretched CH, carbonyl, and alkene. It was consistent with the result of OPLS analysis of LC-MS/MS data, which revealed that based on their VIP and Y-related coefficient value, nystose, 1-kestose, luteolin-3'-7-di-O-glucoside, and 1,3-O-dicaffeoilquinic acid isomers, strongly linked to Yacon's α-glucosidase inhibitor activity. In silico study supported these findings, revealing that the four compounds were potent α-glucosidase inhibitors with docking score in the range of - 100.216 to - 115.657 kcal/mol, which are similar to acarbose (- 115.774 kcal/mol) as a reference drug.

Inhibition of pancreatic elastase in silico and in vitro by Rubus rosifolius leaves extract and its constituents.

OBJECTIVE: Elastases are protease enzymes, which mainly hydrolyze proteins of the connective tissue, so they have a significant impact on human disease. Rubus rosifolius is one of the Rubus species found in Indonesian mountains, and it has potential as an elastase inhibitor. The objective of this research was to examine the in vitro elastase inhibitor activity of R. rosifolius leaves and to dock different ligands of its constituents against target protein of Porcine Pancreatic Elastase (PPE) receptor. METHOD: Dried leaves powder of R. rosifolius was extracted using Soxhlet apparatus with n-hexane, ethyl acetate, and methanol. The extract was evaporated, and in vitro elastase inhibitor activity was determined using PPE with the quercetin used as control positive. Selected nine constituents of R. rosifolius were evaluated on the docking behavior of elastase receptor using Protein-Ligand ANT System (PLANTS) computational software with PPE enzyme with Protein Data Bank (PDB) file 1BRU. RESULT: The methanol extract showed significantly inhibited elastase with IC50 186.13 µg/mL, but ethyl acetate extract showed weak activity, and n-hexane extract did not show any activity. Docking studies and binding free energy calculations and hydrogen bonding with some amino acids revealed that ellagic acid showed the least binding energy for the target enzyme. CONCLUSION: This research has opened new insights into understanding that constituents of R. rosifolius methanol extract are potential inhibitors against elastase, and suggested the active compound is ellagic acid.