Dynamics changes in metabolites and pancreatic lipase inhibitory ability of instant dark tea during liquid-state fermentation by Aspergillus niger.

Instant dark tea (IDT), prepared by liquid-state fermentation using Aspergillus niger, is known for its high theabrownins content and lipid-lowering effect. To explore the impact of fungal fermentation on IDT compositions and its pancreatic lipase inhibitory ability (PLIA), untargeted and targeted metabolomic analysis were applied to track the changes of metabolites over a 9-day fermentation period, and correlation analysis was then conducted between metabolites and PLIA of IDT. There were 54 differential metabolites exhibited significant changes from day 3 to day 5 of fermentation. The concentrations of theabrownins and caffeine increased during fermentation, while phenols and free amino acids decreased. The PLIA of IDT samples significantly increased from day 5 to day 9 of fermentation. Theabrownins not only positively correlated with the PLIA but also exhibited a high inhibition rate. These findings provide a theoretical basis to optimize the production of IDT as functional food ingredient.

Antiobesity Drug Discovery Research: In vitro Models for Shortening the Drug Discovery Pipeline.

Obesity is a growing global health problem, leading to various chronic diseases. Despite standard treatment options, the prevalence of obesity continues to rise, emphasizing the need for new drugs. in vitro methods of drug discovery research provide a time and cost-saving platform to identify new antiobesity drugs. The review covers various aspects of obesity and drug discovery research using in vitro models. Besides discussing causes, diagnosis, prevention, and treatment, the review focuses on the advantages and limitations of in vitro studies and exhaustively covers models based on enzymes and cell lines from different animal species and humans. In contrast to conventional in vivo animal investigations, in vitro preclinical tests using enzyme- and cell line-based assays provide several advantages in development of antiobesity drugs. These methods are quick, affordable, and provide high-throughput screening. They can also yield insightful information about drug-target interactions, modes of action, and toxicity profiles. By shedding light on the factors that lead to obesity, in vitro tests can also present a chance for personalized therapy. Technology will continue to evolve, leading to the creation of more precise and trustworthy in vitro assays, which will become more and more crucial in the search for novel antiobesity medications.

Synergistic effects of Momordica charantia, Nigella sativa, and Anethum graveolens on metabolic syndrome targets: In vitro enzyme inhibition and in silico analyses.

Momordica charantia, Nigella sativa, and Anethum graveolens are established medicinal plants possessing noted anti-diabetic and anti-obesity properties. However, the molecular mechanisms underscoring their inhibitory effects on pancreatic lipase, α-glucosidase, and HMG-CoA reductase remain unexplored. This study aimed to elucidate the efficacy of various NS, MC, and AG blends in modulating the enzymatic activity of pancreatic lipase, HMG-CoA reductase, and a-glucosidase, utilizing an integrative approach combining in vitro assessments and molecular modeling techniques. A factorial design matrix generated eight distinct concentration combinations of NS, MC, and AG, subsequently subjected to in vitro enzyme inhibition assays. Molecular docking analyses using AutoDock Vina, molecular dynamics simulations, MMPBSA calculations, and principal component analysis, were executed with Gromacs to discern the interaction dynamics between the compounds and target enzymes. A formulation comprising NS:MC:AG at a 215:50:35 µg/mL ratio yielded significant inhibition of pancreatic lipase (IC50: 74.26 ± 4.27 µg/mL). Moreover, a concentration combination of 215:80:35 µg/mL effectively inhibited both α-glucosidase (IC50: 66.09 ± 3.98 µg/mL) and HMGCR (IC50: 129.03 µg/mL). Notably, MC-derived compounds exhibited superior binding affinity towards all three enzymes, compared to their reference molecules, with diosgenin, Momordicoside I, and diosgenin displaying binding affinities of -11.0, -8.8, and -7.9 kcal/mol with active site residues of pancreatic lipase, α-glucosidase, and HMGCR, respectively. Further, 100 ns molecular dynamics simulations revealed the formation and stabilization of non-bonded interactions between the compounds and the enzymes' active site residues. Through a synergistic application of in vitro and molecular modeling methodologies, this study substantiated the potent inhibitory activity of the NS:MC:AG blend (at a ratio of 215:80:35 µg/mL) and specific MC compounds against pancreatic lipase, α-glucosidase, and HMGCR. These findings provide invaluable insights into the molecular underpinnings of these medicinal plants' anti-diabetic and anti-obesity effects and may guide future therapeutic development.

Combinatorial preparation and structural characterization of anthocyanins and aglycones from Purple-heart Radish for evaluation of physicochemical stability and pancreatic lipase inhibitory activity.

In this study, an efficient approach to preparation of different anthocyanins from Purple-heart Radish was developed by combining microwave-assisted extraction (MAE), macroporous resin purification (MRP) and ultrasound-assisted acid hydrolysis (UAAH) for evaluation of physicochemical stability and pancreatic lipase (PL) inhibitory activity. By optimization of MAE, MRP and UAAH processes, the anthocyanins reached the yield of 6.081 ± 0.106 mg/g, the purity of 78.54 ± 0.62 % (w/w) and the content of 76.29 ± 1.31 % (w/w), respectively. With high-resolution UHPLC-Q-Orbitrap/MS, 15 anthocyanins were identified as pelargonins with diverse glucosides and confirmed by pelargonidin standard. By glycosylation, pelargonins exhibited higher stability in different pH, temperature, light, metal ions environments than that of pelargonidin. However, PL inhibitory assay, kinetic analysis and molecular docking demonstrated that pelargonidin had higher PL inhibitory activity than pelargonins even though with similar binding sites and a dose-effect relationship. The above results revealed that the effect of glycosylation and deglycosylation on PL inhibitory activity and physicochemical stability.

Applications of Saponin Extract from Asparagus Roots as Functional Ingredient.

When replanting an asparagus field, the roots of the previous crop are crushed and incorporated into the soil, creating problems of autotoxicity and fungal infections. Asparagus roots can be considered as a valuable byproduct, since they are very rich in saponins (3-6%), compounds currently considered as bio-emulsifiers. The objective is to evaluate the emulsifying and foaming capacity of a saponin extract from asparagus roots (ARS) and compare it with other commercial extracts. ARS was obtained using a process patented by our research group. The results have shown that ARS has activity similar to Quillaja extract. Its critical micellar concentration falls between that of Quillaja and Tribulus extracts (0.064, 0.043, and 0.094 g/100 mL, respectively). Both emulsifying and foaming activities are affected by pH, salt, and sucrose to a similar extent as the other extracts. Additionally, it has demonstrated an inhibitory effect on pancreatic lipase, which is even better than the other two studied extracts, as indicated by its IC50 value (0.7887, 1.6366, and 2.0107 mg/mL for asparagus, Quillaja, and Tribulus, respectively). These results suggest that ARS could serve as a natural emulsifying/foaming agent for healthier and safer food products and as a potential aid in treatments for obesity and hyperlipidemia.

Design, synthesis, biological evaluation and molecular docking of alkoxyaurones as potent pancreatic lipase inhibitors.

Aurones are a minor subgroup of flavonoids. Unlike other subgroups such as chalcones, flavones, and isoflavones, aurones have not been extensively explored as pancreatic lipase inhibitors. In this work, we studied the pancreatic lipase inhibitory potency of synthetic aurone derivatives. Thirty-six compounds belonging to four series (4,6-dihydroxyaurone, 6-hydroxyaurone, 4,6-dialkoxyaurone, and 6-alkoxyaurone) were designed and synthesized. Their in vitro inhibitory activities were determined by spectrophotometric assay in comparison with quercetin and orlistat. Alkoxyaurone derivatives with long-chain (6-10 carbons) alkoxy substituents showed greater potency. Of them, 4,6-dialkoxyaurone 8 displayed the highest activity against pancreatic lipase (IC50 of 1.945 ± 0.520 µM) relative to quercetin (IC50 of 86.98 ± 3.859 µM) and orlistat (IC50 of 0.0334 ± 0.0015 µM). Fluorescence quenching measurement confirmed the affinity of alkoxyaurone derivatives to pancreatic lipase. Kinetic study showed that 8 inhibited lipase through a competitive mechanism (Ki of 1.288 ± 0.282 µM). Molecular docking results clarified the role of long-chain substituents on ring A in interacting with the hydrophobic pockets and pushing the inhibitor molecule closer to the catalytic triad. The findings in this study may contribute to the development of better pancreatic lipase inhibitors with aurone structure.

Canary Seed (Phalaris canariensis L.) Peptides Prevent Obesity and Glucose Intolerance in Mice Fed a Western Diet.

Previous research showed that canary seed (Phalaris canariensis L.) peptides (CSP) possess robust in vitro antiobesity properties via inhibition of pancreatic lipase (PL). Nevertheless, no studies have yet explored their antiobesity properties in vivo. Consequently, we investigated the effects of CSP in C57BL/6J mice under a Western diet (WD). Mice were assigned into groups and fed a normal diet (ND) or a WD accompanied by an oral dose of CSP (250 or 500 mg/kg/day), orlistat (40 mg/kg/day), or distilled water. The results showed that consuming CSP can provide metabolic benefits, including preventing weight gain by up to 20%, increasing glucose tolerance, and reducing insulin, leptin, and LDL/VLDL levels in plasma. Conversely, total ghrelin was unaffected by CSP-500, but decreased by CSP-250, and amplified by orlistat. Surprisingly, CSP-250 was more effective in preventing weight gain and promoting satiety than CSP-500. Parallel to this, protein absorption in CSP-500 was decreased, supported by a rise in fecal crude protein (+3.5%). Similarly, fecal fat was increased by orlistat (38%) and was unaffected by CSP-250 (3.0%) and CSP (3.0%), comparatively to WD (2.5%). Despite this, both CSP treatments were equally effective in decreasing hepatic steatosis and avoiding hyperlipidemia. Furthermore, the enzymatic analysis showed that CSP-PL complexes dissociated faster (15 min) than orlistat-PL complexes (41 min). Lastly, CSP did not affect expression of hepatic lipid oxidation genes ACO and PPAR-α, but reduced the expression of the hydrolase gene LPL, and lipogenesis related genes FAS and ACC. Taken together, these results suggest that CSP antiobesity mechanism relies on lipid metabolism retardation to increase fat transit time and subsequently suppress hunger.

Pancreatic lipase inhibitory effects of peptides derived from sesame proteins: In silico and in vitro analyses.

Pancreatic lipase (PL) is the main digestive enzyme that is responsible for breaking triglycerides into smaller components for absorption. Inhibition of PL can effectively reduce triglyceride absorption, helping to prevent the development of obesity. The objective of this study was to investigate the PL inhibitory activity of peptides derived from sesame (Sesamum indicum L.) in silico and in vitro. In silico proteolysis of sesame proteins with pepsin, trypsin and chymotrypsin was performed with ExPASy PeptideCutter. Six peptides (TF, EW, QWM, NIF, AGY and PIF) were screened out by PeptideRanker, SwissADME and AutoDock. Molecular docking analysis showed that these six peptides could interact directly with Phe77, His151, Ser152, Phe215 and His263 at the key sites of PL. The six peptides were further synthesized to verify their PL-inhibitory effects in vitro, and TF, EW, QWM, NIF and AGY exhibited inhibitory activity on PL with IC50 values of 751 ± 75, 907 ± 91, 986 ± 170, 1044 ± 179 and 1183 ± 179 µM, respectively. Inhibitory kinetics indicated that TF, QWM and NIF were mixed-type inhibitors of PL, while EW and AGY were uncompetitive inhibitors. Our results suggest that peptides from sesame could potentially inhibit the activity of PL.

Chemical constituents from mushroom Geoglossum fallax and their bioactive activities.

Chemical investigation on the fruiting bodies of the mushroom Geoglossum fallax led to the isolation of two carboxamides, geoglamides A and B (1 and 2), one meroterpenoid, geoglol A (3), together with seven known metabolites (4-10). Their structures were identified by spectroscopic analyses as well as ECD calculations. Compounds 1 and 2 are rare long chain fatty amides containing a 4-oxo-1,4-dihydropyridine moiety, while compound 3 is a rare C6-C5 type meroterpenoid. Compound 1 displayed cytotoxic activity against human MCF-7 cells with an IC50 value of 25.9 ± 0.51 µM. compounds 2 and 8 showed weak pancreatic lipase inhibition activity. To our best knowledge, this is the first report of the chemical constituents in the genera Geoglossum and their bioactive activity.

Resin glycoside extracts from Ipomoea aquatica retard lipid digestibility of high-fat food in vitro.

Due to the considerable increase in the prevalence of obesity, there is an increased interest in developing safe and effective anti-obesity treatments from fruits and vegetables. In this study, Ipomoea aquatica, commonly known as Kang Kong in Southeast Asia was first reported to contain potent pancreatic lipase (PL) inhibitors due to resin glycosides (RG). Ipomoea aquatica extract demonstrated a dose-dependent inhibitory activity against PL with an Orlistat equivalent (OE) value of 6.86 ± 0.51 × 10-4. In vitro lipolysis study showed that consuming RG in tandem with high-fat food (butter & salad dressing) was effective in delaying enzymatic fat digestion by inhibiting PL. Pre-incubation of PL with RG extract before substrate addition also significantly enhanced their inhibitory activity. However, RG was unstable when subjected to high heat treatments (90 °C). Overall, these results provided useful knowledge of RG as PL inhibitors for body weight management.

Penicipurate A, a new polyketide derivative from the endophytic fungus Penicillium purpurogenum.

A new polyketide derivative containing a 3-hydroxydecanoic acid ester moiety, penicipurate A (1), was purified from the solid cultures of the fungus Penicillium purpurogenum, a fungal strain endophytic in the leaves of Edgeworthia chrysantha. The structure of 1 was established by spectroscopic methods, including UV, IR, HRESIMS, 1D, and 2D NMR and 13C NMR chemical shifts calculations coupled with DP4+ analysis, as well as the chemical degradation method. Compound 1 showed moderate inhibitory activity against pancreatic lipase (PL) with an IC50 value of 9.61 ± 1.42 µM.

Honeysuckle Berry (Lonicera caerulea L.) Inhibits Lipase Activity and Modulates the Gut Microbiota in High-Fat Diet-Fed Mice.

Honeysuckle berry (HB, Lonicera caerulea L.) is an oriental herbal medicine reported to have beneficial effects on metabolic disorders, such as obesity and non-alcoholic fatty liver disease. The fruit part of HB is rich in anthocyanin, a type of polyphenol. Most studies credit the antioxidant and anti-inflammatory properties of HB as the mechanisms of its effectiveness. This study investigated the inhibitory effects of HB on lipase using an in vitro assay and the modulatory effect of HB on gut microbiota in high-fat diet (HFD)-fed mice. HB inhibited pancreatic lipase activity with IC50 values of approximately 0.47 mg/mL. The fecal triglyceride (TG) levels were higher from the HFD of the HB-fed mice than they were for the control mice. Moreover, the fecal microbiota from the HFD of the HB-fed mice had relatively lower Firmicutes and higher Bacteroidetes than that from the HFD-only mice. These results suggest that HB modulates gut microbiota composition, which may contribute to body fat reduction. Hence, HB could present a useful agent for treating metabolic diseases through lower TG uptake and the regulation of gut microflora.

Evaluation of in vitro lipid-lowering properties of ‘Saba’ banana [Musa acuminata x balbisiana (BBB group) ‘Saba’] peel pectin from different extraction methods

@#Introduction: This study was conducted to investigate the in-vitro lipid-lowering properties of ‘Saba’ banana peel pectin (SBP) extracted using three methods for its possible use as a dietary fibre ingredient. Methods: Pectin from ‘Saba’ banana peels were extracted using acid extraction (citric acid), enzymatic extraction (cellulase), and microwave-assisted extraction. In-vitro lipid-lowering assays were performed using spectrophotometry for pancreatic lipase inhibition and cholesterol binding, while liquid chromatography was used for bile acid-binding capacity. Results: Results revealed that all SBPs were not able to inhibit pancreatic lipase activity. However, all SBPs can notably bind to cholesterol and bile acids, taurocholate, and glycocholate. Acid-extracted pectin had the highest binding capacity to cholesterol (51.36%–55.07%) and glycocholate (27.37%), whereas all SBPs were similarly bound to taurocholate. Conclusion: The results of this study showed that acidextracted SBPs can significantly bind to cholesterol and bile acids, glycocholate and taurocholate, thereby indicating a possible reduction in lipid metabolism.

Screening of in vitro and in silico α-amylase, α-glucosidase, and lipase inhibitory activity of oxyprenylated natural compounds and semisynthetic derivatives.

Metabolic syndrome has several characteristic manifestations, including insulin resistance and dyslipidaemia, that demand therapeutic approaches, such as the inhibition of enzymes involved in nutrient absorption and digestion.This study aimed to evaluate the potential pharmacological use of natural compounds widespread in the plant kingdom and their semisynthetic compounds against target enzymes. Twenty-three oxyprenylated natural compoundswere investigated for their ability to inhibit α-amylase, α-glucosidase, and pancreatic lipase enzymes by in vitro assays. Moreover, in silico molecular docking was performed to analyse their binding capabilities into 3D structures. Farnesyloxyferulic acid, geranyloxyvanillic acid, nelumal A, and geranyloxyferulic acid showed the highest inhibition activity in all three in vitro enzyme assays. Moreover, in silico molecular docking of these four compounds was used to analyse their possible binding in 3D structures of the investigated enzymes. The results indicate that these compounds have considerable therapeutic potential for the treatment of metabolic syndrome, and further studies are warranted for their pharmacological development.

Paeonone A, a novel nonanortriterpenoid from the roots of Paeonia lactiflora.

Paeonone A (1), a unique nonanortriterpenoid, and a new octanortriterpenoid, paeonone B (2), were isolated from the roots of Paeonia lactiflora, together with a known analogue, palbinone (3). Paeonone A (1) is the first example of naturally occurring nonanortriterpenoid with a diketo acid group. Extensive NMR and HRESIMS experiments were applied to identify the structures of 1 and 2, and their absolute configurations were solved by single-crystal X-ray diffraction and ECD data. Biological properties of 1-3 were explored against pancreatic lipase and cancer cell lines.

Antioxidant, Pancreatic Lipase Inhibitory, and Tyrosinase Inhibitory Activities of Extracts of the Invasive Plant Spartina anglica (Cord-Grass).

Since 2016, the invasive halophyte Spartina anglica has been colonizing mudflats along the western coast of South Korea. In order to minimize costs on S. anglica expansion management and waste-treatment of collected biomass, the potential application of the collected biomass of S. anglica was investigated. Ethanolic extracts and subfractions thereof (hexanes, methylene chloride, ethyl acetate, 1-butanol, and water-soluble) of the aerial and belowground parts of S. anglica showed free radical-scavenging [2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS)], tyrosinase inhibitory, and pancreatic lipase inhibitory activities. An ethyl acetate fraction derived from aerial parts (EA-a) showed the most potent radical-scavenging and pancreatic lipase inhibitory activities, whereas tyrosinase inhibition was mainly observed in the methylene chloride soluble fractions (MC-bg) and other lipophilic fractions (ethyl acetate and hexanes layers) obtained from belowground parts. The major EA-a compound isolated and identified was 1,3-di-O-trans-feruloyl quinic acid (1) based on spectroscopic analysis, whereas the two major MC-bg compounds were identified as p-hydroxybenzaldehyde (2) and N-trans-feruloyltyramine (3). Compounds 1 and 3 scavenged both DPPH and ABTS radicals, whereas 1 and 2 inhibited pancreatic lipase activity. These results indicate that extracts and fractions of S. anglica have antioxidant, anti-obesity, and whitening properties with potential pharmaceutical, cosmeceutical, and functional food applications.

In-vitro functional efficacy of extracts from Phyllanthus emblica, Eucalyptus globulus, Tinospora cordifolia as pancreatic lipase inhibitor and source of anti-oxidant in goat meat nuggets.

The present study was aimed to evaluate the functional efficacy of plant extracts as a source of pancreatic lipase inhibitor and antioxidant in goat meat nuggets to address the fat paradox issue of red meat. The PPLIA, antioxidant potential, and resistance against fat digestion were in the order ofPhyllanthus emblica > Eucalyptus globulus > Tinospora cordifolia.PPL inhibition activities of water and ethanolic extracts fromPhyllanthus emblicausing DNPB and Triolein as substrate were 63.76, 67.94 and 56.17 and 64.36 percent respectively whereas, TPC, DPPH RSA, FRPA were 40.82 and 59.52 (mgGAE/g), 54.89 and 59.84 (percent), 1.26 and 1.61 (OD) respectively. The average diameter of fat globules in digest was maximum (8.91 µm) withPhyllanthus emblicafruits extracts whereas; TBARs (0.347 mg MDA/Kg) and FFA (4.47 µg/g) values were lowest. This study showed that extracts from plants can act as a promising natural alternative in the development of healthy meat products.

Dihydrochalcone C-glycosides from Averrhoa carambola leaves.

Ten undescribed dihydrochalcone C-glycosides, carambolasides R1‒R3, S1, S2, T1‒T3, 3-hydroxycarambolaside T1, and 3-hydroxycarambolaside P were isolated along with carambolasides I and P from the leaves of Averrhoa carambola L. (Oxalidaceae). Their structures were determined by spectroscopic and chemical methods. Among them, carambolasides P, T1, T2, and I with contents of 22.78, 14.39, 4.93, and 1.87 mg g-1 dry wt., respectively, were shown to be abundant in the leaves by HPLC analysis. All the compounds showed more potent ABTS radical cation scavenging activity than l-ascorbic acid. 3-Hydroxycarambolaside T1 and 3-hydroxycarambolaside P also demonstrated moderate DPPH radical scavenging activity. Further, carambolaside R3, 3-hydroxycarambolaside T1, and 3-hydroxycarambolaside P exhibited weak in vitro porcine pancreatic lipase inhibitory activity.

Cyanidin and cyanidin-3-glucoside derived from Vigna unguiculata act as noncompetitive inhibitors of pancreatic lipase.

The consumption of legumes positively correlated with the reduction of body weight. In the present study, we identified and evaluated pancreatic lipase inhibitors from Vigna unguiculata and unraveled their mode of inhibition. The highly sensitive fluorometric method was adopted to access the pancreatic lipase activity and the ethanolic extract of Vigna unguiculata showed the maximum inhibition (IC50 of 15.2 µg/ml). Cyanidin and cyanidin-3-glucoside are the major anthocyanins observed in Vigna unguiculata. The IC50 value of cyanidin was 28.29 µM which was 6.5-fold higher than the cyanidin-3-glucoside (188.28 µM). We determined an apparent Ki of 27.28 µM for cyanidin and cyanidin-3-glucoside (88.97 µM) with noncompetitive inhibition. Collectively, these results suggest that the glycosylation of the anthocyanidins significantly reduces lipase inhibition. The noncompetitive inhibition of pancreatic lipase by Vigna unguiculata anthocyanins may exert significant pharmacological activities toward obesity complications by calorie restriction. PRACTICAL APPLICATIONS: The results of this study emphasize the importance of legumes in our diet to combat obesity-related complications. Consumption of legumes minimizes fat absorption by inhibiting the action of the fat-digesting enzyme.

Acyl atractyligenin and carboxyatractyligenin glycosides from Antennaria rosea subsp. confinis.

Eight previously undescribed acyl atractyligenin and carboxyatractyligenin glycosides were isolated from whole Antennaria rosea subsp. confinis (Greene) R. J. Bayer (Compositae) [syn. Leontopodium leontopodioides (Willd.) Beauv. (Asteraceae)] plants and their structures were determined by spectroscopic and chemical methods. The compounds were trivially named leontopodiosides F-M. Seven of the compounds showed potent in vitro inhibitory activity toward pancreatic lipase with IC50 values ranging from 3.4 to 52.5 µM, suggesting that they participate in the previously observed effect this plant has in reducing triglyceride absorption in rats.