Pancreatic Lipase Inhibition Assay Research Articles

Targeted Screening of Curcumin Derivatives as Pancreatic Lipase Inhibitors Using Computer-Aided Drug Design.

Curcumin has demonstrated promising preclinical antiobesity effects, but its low bioavailability makes it difficult to exert its full effect at a suitable dose. The objective of this study was to screen curcumin derivatives with enhanced bioavailability and lipid-lowering activity under the guidance of computer-aided drug design (CADD). CAAD was used to perform virtual assays on curcumin derivatives to assess their pharmacokinetic properties and effects on pancreatic lipase activity. Subsequently, 19 curcumin derivatives containing 5 skeletons were synthesized to confirm the above virtual assay. The in vitro pancreatic lipase inhibition assay was employed to determine the half-maximal inhibitory concentration (IC50) of these 19 curcumin derivatives. Based on CADD analysis and in vitro pancreatic lipase inhibition, 2 curcumin derivatives outperformed curcumin in both aspects. Microscale thermophoresis (MST) experiments were employed to assess the binding equilibrium constants (K d) of the aforementioned 2 curcumin derivatives, curcumin, and the positive control drug with pancreatic lipase. Through virtual screening utilizing a chemoinformatics database and molecular docking, 6 derivatives of curcumin demonstrated superior solubility, absorption, and pancreatic lipase inhibitory activity compared to curcumin. The IC50 value for 1,7-bis(4-hydroxyphenyl)heptane-3,5-dione (C4), which displayed the most effective inhibitory effect, was 42.83 μM, while the IC50 value for 1,7-bis(4-hydroxy-3-methoxyphenyl)heptane-3,5-dione (C6) was 98.62 μM. On the other hand, the IC50 value for curcumin was 142.24 μM. The MST experiment results indicated that the K d values of C4, C6, and curcumin were 2.91, 18.20, and 23.53 μM, respectively. The results of the activity assays exhibited a relatively high degree of concordance with the outcomes yielded by CADD screening. Under the guidance of CADD, the targeted screening of curcumin derivatives with excellent properties in this study exhibited high-efficiency and low-cost benefits.

Flavonoid extracts of Citrus aurantium L. var. amara Engl. Promote browning of white adipose tissue in high-fat diet-induced mice

Ethnopharmacological relevanceObesity has become a public burden worldwide due to its booming incidence and various complications, and browning of white adipose tissue (WAT) is recognized as a hopeful strategy to combat it. Blossom of Citrus aurantium L. var. amara Engl. (CAVA) is a popular folk medicine and dietary supplement used for relieving dyspepsia, which is recorded in the Chinese Materia Medica. Our previous study showed that blossom of CAVA had anti-obesity potential, while its role in browning of WAT was still unclear. Aim of the studyThis study aimed to characterize the constituents in flavonoids from blossom of CAVA (CAVAF) and to clarify the anti-obesity capacities especially the effects on browning of WAT. Materials and methodsGradient ethanol eluents from blossom of CAVA were obtained by AB-8 macroporous resin. 3T3-L1 cells and pancreatic lipase inhibition assay were employed to investigate the potential anti-obesity effects in vitro. HPLC and UPLC/MS assays were performed to characterize the chemical profiles of different eluents. Network pharmacology and molecular docking assays were used to reveal potential anti-obesity targets. Furthermore, high-fat diet (HFD)-induced mice were constructed to explore the anti-obesity actions and mechanisms in vivo. Results30% ethanol eluents with high flavonoid content and great inhibition on proliferation of 3T3-L1 preadipocytes and pancreatic lipase activity were regarded as CAVAF. 19 compounds were identified in CAVAF. Network pharmacology analysis demonstrated that AMPK and PPARα were potential targets for CAVAF in alleviating obesity. Animal studies demonstrated that CAVAF intervention significantly decreased the body weight, WAT weight, serum TG, TC and LDL-C levels in HFD-fed obese mice. HFD-induced insulin resistance and morphological changes in WAT and brown adipose tissue were also markedly attenuated by CAVAF treatment. CAVAF supplementation potently inhibited iWAT inflammation by regulating IL-6, IL-1β, TNF-α and IL-10 mRNA expression in iWAT of mice. Furthermore, the gene expression levels of thermogenic markers including Cyto C, ATP synthesis, Cidea, Cox8b and especially UCP1 in iWAT of mice were significantly up-regulated by CAVAF administration. CAVAF intervention also markedly increased the expression levels of PRDM16, PGC-1α, SIRT1, AMPK-α1, PPARα and PPARγ mRNA in iWAT of mice. ConclusionCAVAF treatment significantly promoted browning of WAT in HFD-fed mice. These results suggested that flavonoid extracts from blossom of CAVA were probably promising candidates for the treatment of obesity.

Isoflavone-Rich Extract of Trifolium resupinatum: Anti-obesity Attributes with In Silico Investigation of Its Constituents

Trifolium resupinatum L., Fabaceae, aqueous methanol leaf extract was selected to mitigate some obesity-associated risk factors to validate the possibility of further developing herbal drugs. Chromatography and spectrophotometric techniques verified 14 phenolics, five of which were first isolated from the plant and identified as 6''-O-acetyl ononin, genistin, daidzin, sissotrin, and astragalin. Further phytochemical characterization was performed via liquid chromatography-electrospray ionization-mass spectrometry assisted by a spectral similarity molecular network. In total, 81 metabolites were tentatively annotated including 69 species-first dereplications. Two major isolates (formononetin and pseudobaptigenin) were selected along with the investigated extract for an in vitro pancreatic lipase inhibition assay. They showed notable effects with IC50 values (µg/ml): 47.2 ± 1.1, 112.8 ± 1.23, and 471.32 ± 0.8, respectively, incomparable to orlistat (23.8 ± 0.64). Preliminary in vivo assay (25 mg/kg extract, daily, 8 weeks) displayed weight loss interest and promising advancement of serum triacylglycerides, total cholesterol, and glucose levels. Molecular docking studies confirmed the promising binding score of formononetin and pseudobaptigenin near the active sites and highlighted the affinity of other isolates to the lipase enzyme. Several isolates passed Lipinski’s law of the drug-likeness test, whereas SwissADME radar displayed that all constituents fall within the acceptable bioavailability zone. Therefore, the combination of flavonoids, especially isoflavones, could be regarded as drug-like agents for protection against obesity-induced metabolic complaints.Graphical

Pancreatic Lipase inhibition assay of various extracts of leaves of Murraya Koenigii in southern areas of Goa

The objective of the study was to assess the lipase inhibitory activities of chloroformic, methanolic and aqueous extracts from the commonly available Murraya koenigii (L.) Spreng leaves(Rutaceae) in southern villages of Goa, for potential use in the treatment of obesity. Extracs of the leaves of this plant were evaluated for lipase inhibitory activity using porcine pancreatic lipase (PPL: triacylglycerol lipase) and p-nitrophenyl butyrate in an in vitro assay. Among the three extracts screened, chloroformic extract exhibited the highest pancreatic lipase inhibitory activity of 53.42%, followed by methanolic extract (51.88%) and aqueous extract (36.42%), respectively. Chloroformic extract has not been screened for its pancreatic lipase inhibition assay. All the Crude extracts of leaves of Murraya koenigii (L.) Spreng leaves (Rutaceae) have potential as pancreatic lipase inhibitory agents. . Chloroformic extract was found to be most effective and hence can be used as a potent anti-obesity agent to combat hyperlipidemia.

Development and evaluation of anti-reflux functional-oral suspension raft composed of sodium alginate-mung bean protein complex

This study aimed to develop a sodium alginate (Na alginate) and mung bean protein (MBP) raft complex to improve gastric reflux symptoms. Na alginate and MBP complexes with different ratios (1:1, 2:1, and 3:1, respectively) were used for raft formulations through a wet Maillard reaction. Structural properties of raft strength, reflux resistance, intrinsic fluorescence emission spectroscopy, and Fourier transform infrared spectroscopy (FTIR) were investigated for rafts. The suspension 1:1 Na alginate/MBP with 0 h Maillard reaction time exhibited the lowest sedimentation volume among the suspensions. In contrast, 3:1 Na alginate/MBP with 6 h Maillard reaction time showed the highest sedimentation volume. Based on the results, the 3:1 Na alginate/MBP rafts had the best results, and the results were within acceptable limits. Functional properties, including antioxidant properties, the Helicobacter pylori inhibition assay, the pancreatic lipase inhibition assay, and angiotensin-converting enzyme (ACE) inhibition, were investigated for rafts. The Na alginate/MBP raft has similar characteristics to Gaviscon syrup and can be used for obesity, Helicobacter pylori infection, high blood pressure, and gastric reflux.

Potential lipase inhibitor from underutilized part of Andrographis paniculata: Targeted isolation and mechanism of inhibition

Pancreatic lipase (PL) plays a vital role in lipid metabolism-associated disorders, especially obesity. The present study demonstrates the targeted isolation of biologically active compounds from Andrographis paniculata root with dietary fat absorption capacity for obesity management. The HPLC-based micro-fractionation and off-line in-vitro pancreatic lipase inhibition assay have resulted in potential phytochemicals with IC50 value 4.46–21.76 µg/mL. 7-O-methyl-dihydro wogonin (MDHW) was the most potent with IC50 = 14.86 ± 0.19 μM than reported flavanones. The MDHW has also exhibited a synergistic effect with orlistat at a concentration lower than 14.86 μM. MDHW acts as a competitive inhibitor via energy transfer from PL. The binding interaction of MDHW with PL was hydrophobic (Ka=1.40 ×103 L/mol). FT-IR studies showed the stable and compact MDHW-PL complex as the percentage of β -turn, β-antiparallel and random coil increased, while the portion of the α-helix, β-sheet decreased. Additionally, in-silico studies have also confirmed the non-polar interaction of MDHW with the protein's active site (PBD:1ETH) by spontaneous H-bonds. Results suggested that MDHW isolated from A. paniculata root showed high PL inhibitory activity and binding affinity (Ki =7.4 µM), promising a potential anti-obesity agent for pharmaceutical and food industry applications.

Formulation of plant-based yoghurt from soybean and quinoa and evaluation of physicochemical, rheological, sensory and functional properties

The aim of this study was to develop a novel plant-based yoghurt with high sensory acceptance and nutrition value. Soybean and quinoa were used as the substrates, and all formulations with different substrate ratios were inoculated with lactic acid bacteria (LAB). Fermentation modified the proximate composition, and a higher content of soybean accentuated the growth of LAB. Formulated yoghurts presented a weak post-acidification, and the viability of LAB was higher than 106−7 CFU/g during a 21 days storage. Quinoa addition significantly improved the water holding capacity (WHC) and decreased the yellowness, and an increase of lightness and a decrease of WHC occurred during the storage. Compared with the plain substrate yoghurt, mixed formulation had no significant impact on textual parameters, whereas increased the liking scores in sensory evaluation. Furthermore, fermentation improved the total phenolic content (TPC), which showed a dose-dependent relationship between soybean content. Results obtained from DPPH, ABTS, hydroxyl radical scavenging assay, FRAP assay, and α-amylase, α-glucosidase and pancreatic lipase inhibition assay showed that fermentation enhanced the antioxidant capacities and digestive enzyme inhibitory activities. TPC increase and phenolic biotransformation may explain the enhancement of the nutritive values. Based on the sensory and function evaluation, the ratio of quinoa against soybean at 8:2 and 2:8 presented the best formulations.

In Vitro Pancreatic Lipase Inhibition by Marine Fungi Purpureocillium lilacinum Associated with Stylissa sp. Sponge as Anti-obesity Agent

This study aimed to evaluate the potential of marine fungus Purpureocillium lilacinum isolated from an Indonesian marine sponge Stylissa sp. as an anti-obesity agent through pancreatic lipase inhibition assay. The fungus was identified as P. lilacinum through morphological and molecular characteristics. The fungal extract’s inhibition activity and kinetics were evaluated using spectrophotometry and Lineweaver-Burk plots. Ethyl acetate and butanol were used for extraction. Both extracts showed pancreatic lipase inhibition in a concentration-dependent manner. Both crude extracts were then fractionated once. All fractionated extracts showed inhibitory activity above 50%, with the highest activity found in fraction 5 of ethyl acetate at 93.41% inhibition. The best fractionated extract had an IC50value of 220.60 µg.mL-1. The most active fraction of P. lilacinum had a competitive-type inhibitor behavior as shown by the value of Vmax not significantly changing from 388.80 to 382.62 mM pNP.min-1, and the Michaelis-Menten constant (KM) increased from 2.02 to 5.47 mM in the presence of 500 µg.mL-1 fractionated extract. Metabolite identification with LC-MS/MS QTOF suggested that galangin, kaempferol, and quercetin were responsible for the observed lipase inhibition.

Assessment of Antioxidant and Pancreatic Lipase Activity of Smilax zeylanica Roots: An in vitro Analysis

Objectives: It is well established fact that the phytochemicals with phenolic and flavonoids origin have antioxidant property. Thus in this study it was aimed to determine the in vitro antioxidant potential of various extracts of Smilax zeylanica L. stems. Methods: The Photochemical screening was done following the standard procedures. The Antioxidant activity was tested using several in vitro assays, viz., 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The Total phenol and flavonoid contents were determined by colorimetric method. Further porcine lipase activity was determined. Results: The phytochemical screening revealed the presence of phenols and flavonoids the extracts. The Methanolic extract showed significant antioxidant potential than other extracts in all assays. The IC50 value of cyclo hexane extract , ethyl acetate extract and methanol extract of Smilax zeylanica was 29.14 ± 0.39 μ g/mL, 78.41 ±5 .53 μg /mL and 120.30 ±3 .32 μg /mL respectively in DPPH assay. Porcine pancreatic lipase inhibition assay lshowed dose-dependent effect. The % Inhibition of cyclohexane extract (SZH) was 37.52% 54.44% and 77.07% at the highest dose 200 μg/ ml. The highest inhibition was obtained in the methanolic extract at 200 μg/ ml conc which was close to the standard orlistat i.e. 82.39 at 200 μg/ml. Conclusion: The methanolic extracts showed potential pancreatic lipase inhibition.

Screening of Anti-Lipase Components of Artemisia argyi Leaves Based on Spectrum-Effect Relationships and HPLC-MS/MS

Pancreatic lipase is a key lipase for triacylglyceride digestion and absorption, which is recognized as a promising target for treatment of metabolic disorders. Natural phytochemicals are hopeful sources for pancreatic lipase inhibitors. The leaves of Artemisia argyi H.Lév. and Vaniot (AL) is commonly used as herbal medicine or food supplement in China and other Asian countries for hundreds of years. AL mainly contains essential oils, phenolic acids, flavonoids and terpenoids, which exhibit many pharmacological activities such as antioxidant, anti-inflammatory, antimicrobial, analgetic, anti-cancer, anti-diabetes and immunomodulatory effects. However, the anti-lipase activity of AL was lack of study and the investigation of anti-lipase ingredients from AL was also insufficient. In the present study, the anti-lipase activity of AL was evaluated in vitro and the potentially pancreatic lipase inhibitors of AL were investigated. High performance liquid chromatography was used to establish fingerprints of AL samples, and fifteen peaks were selected. The anti-lipase activities of AL samples were evaluated by a pancreatic lipase inhibition assay. Then, the spectrum-effect relationships between fingerprints and pancreatic lipase inhibitory activities were investigated to identify the anti-lipase constitutes in AL. As the results, four caffeoylquinic acids, which were identified as neochlorogenic acid, chlorogenic acid, isochlorogenic acid B, and isochlorogenic acid A by high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, were selected as potential pancreatic lipase inhibitors in AL. Moreover, anti-lipase activity assessment and molecular docking study of the four compounds were performed to validate the potential lipase inhibitors in AL. The results revealed that the four caffeoylquinic acids in AL as bioactive compounds displayed with anti-lipase activity. The present research provided evidences for the anti-lipase activity of AL, and suggested that some bioactive compounds in AL could be used as lead compounds for discovering of new pancreatic lipase inhibitors.

Identification of novel potent pancreatic lipase inhibitors from Ficus racemosa

Introduction and Aim: Obesity is a disorder of lipid metabolism and continues to be a global problem, ranking fifth for deaths worldwide. It is considered to be main reason for a number of physiological changes that resulting in many metabolic disorders such as cardiovascular diseases, diabetes, musculoskeletal disorders and various cancer types. Obesity develops from long-termed physiological imbalances in energy expenditure.. One of the therapeutic strategies in managing obesity is inhibition of pancreatic lipase, a key enzyme responsible for the digestion of fats and triglycerides. The aim of the present study was to synthesize silver nanoparticles (Ag-NP) from Ficus racemosa fruit extract and to analyze its activity on pancreatic lipase enzyme using spectroscopic, in vitro and in silico methods.
 Materials and Methods: The phytoconstituent were separated in Ficus racemosa fruit extract by thin layer chromatography technique. The bioactive compound was also identified by gas chromatography-mass spectrometry analysis. Pancreatic lipase inhibition assay was performed in vitro and confirmed by molecular docking analysis. 
 Results: The F. racemosa fruit showed the highest pancreatic lipase inhibitory activity. Molecular docking studies also exhibited good binding affinity of compounds (Diethyl Phthalate) with pancreatic lipase enzymes. 
 Conclusion: Finally, it is concluded that further derivation of such compounds could serve as the new templates for obesity.

Standardization of Juniperus macrocarpa Sibt. & Sm. and Juniperus excelsa M. Bieb. Extracts with Carbohydrate Digestive Enzyme Inhibitory and Antioxidant Activities.

Juniperus species growing in Turkey are used for various medicinal purposes in traditional folk medicine. We aimed to evaluate in-vitro antidiabetic (α-glucosidase and α-amylase inhibition assays), antiobesity (pancreatic lipase inhibition assay), and antioxidant (ABTS and DPPH radical scavenging activities, ferric reducing activity, metal chelating activity, and phosphomolybdenum assay) activities of the extracts obtained from branches, leaves, and fruits of Juniperus macrocarpa and Juniperus excelsa. The branch (IC50 = 67.1 ± 1.7 µg/mL) and leaf ethyl acetate extracts (IC50 = 83.4 ± 0.8 µg/mL) of J. macrocarpa exhibited the strongest activity on the α-glucosidase enzyme. Besides that, J. excelsa leaf methanol extract exerted remarkable α-amylase inhibitory activity (IC50 = 950.1 ± 3.5 µg/mL). Only J. macrocarpa branch and J. excelsa leaf ethyl acetate extract slightly inhibited pancreatic lipase enzyme with 2963.3 ± 736.4 and 2343 ± 557.8 µg/mL IC50 values, respectively. The RP-HPLC-DAD analysis results demonstrated that the more active J. macrocarpa extracts are richer in agathisflavone, amentoflavone, and umbelliferone than J. excelsa extracts. With this study, it is concluded that J. macrocarpa branch and leaf ethyl acetate extracts may be a new source of α-glucosidase enzyme inhibitory activity and agathisflavone, amentoflavone can be used in the standardization of the extracts.

Evaluation of anti-obesity potential of aqueous extract of Achyranthes aspera Linn. in high fat diet induced obese rats

BackgroundObesity, reached epidemic proportions globally is often associated with life threatening comorbidities. The unavailability of safe and effective long term medications for obesity in modern pharmacotherapy forces the scientific community to explore the potential of Ayurvedic traditional healers as they are considered safe and effective.ObjectiveTo explore the anti-obesity potential of aqueous extract of aerial parts of Achyranthes aspera L. (AEAA), a traditional healer in high fat diet (HFD) induced obese rats.MethodsAEAA was prepared and subjected to in-vitro pancreatic lipase inhibition assay and in-vivo anti-obesity studies. For in-vivo studies, HFD fed obese prone Wistar albino rats were divided into five experimental groups (Group II to VI): animals fed with standard pellet chow served as normal control (Group I) while, animals continued with HFD alone served as obese control (Group II); Group III, IV and V were administered AEAA at a dose of 100, 200 and 300 mg/kg b.w. respectively along with HFD; and animals administered orlistat (30 mg/kg bw) along with HFD served as standard control (Group VI). All the drugs were administered orally once a day for a period of 60 days. At the end of the experimental period various physical, biochemical and histopathological observations were made.ResultsIn-vitro studies showed AEAA partially but not significantly inhibited the activity of pancreatic lipase. Data of in-vivo studies revealed, significant reduction in body weights, fat pad weights and organ weights upon AEAA treatment. Elevated levels of glucose, insulin, leptin, lipid profiles and antioxidant status were also brought back to normal.ConclusionThe obtained results clearly suggested that AEAA possess pronounced anti-obesity potential.

Scientific evaluation of anti-obesity potential of aqueous seed kernel extract of Mangifera indica Linn. in high fat diet induced obese rats

BackgroundObesity is a medical condition often associated with chronic diseases such as hyperlipidemia, Type 2 diabetes mellitus, colon cancer, other cardiovascular diseases etc., and is become a major health problem worldwide in recent years. The non-satisfactory long term effectiveness as well as the side effects associated with the modern pharmacotherapy forced the suffering mankind to resort to better, long lasting and safe therapy. At present, the potential of natural products for treating obesity is under exploration. The aim of the studyTo explore the anti-obesity potential of Aqueous extract of seed kernel of Mangifera indica Linn. (AEMI) in high fat diet (HFD) induced obese rats. MethodologyAEMI was subjected to in-vitro pancreatic lipase inhibition assay. For in-vivo studies, after administering HFD for a period of 45 days obese prone Wistar strains of albino rats were divided into five experimental groups (Group II to VI): animals fed with standard pellet chow served as normal control (Group I) while, animals continued with HFD alone served as obese control (Group II); Group III, IV and V were administered AEMI at a dose of 100, 200 and 300 mg/kg bw respectively along with HFD; and animals administered orlistat (30 mg/kg bw) along with HFD served as standard control (Group VI). All the drugs were administered orally once a day for a period of 60 days. At the end of the experimental period various physical and biochemical parameters were analysed. ResultsIn-vitro studies showed AEMI partially inhibited the activity of the pancreatic lipase. Data of in-vivo studies revealed, significant reduction in body weights, fat pad weights and organ weights of AEMI treated animals. Elevated levels of glucose, insulin, leptin and lipid profiles were normalized upon treatment with AEMI. Antioxidant status was also brought back to normal upon AEMI treatment. ConclusionThese results suggest that the aqueous extract of seed kernel of Mangifera indica Linn. Possess pronounced anti-obesity activity and this effect may be due the pancreatic lipase inhibitory activity as well as potent antioxidant nature of therapeutic phytoconstituents present in the selected plant extract in addition to other unrevealed actions.

Phenolic Compounds with Antioxidant Properties from Canola Meal Extracts Inhibit Adipogenesis.

The extraction of phenolic compounds from canola meal produces functional health products and renders the canola meal a more digestible animal feed. The extracted phenolics may have novel bioactivity worth investigation. In this study, several solvents were evaluated for their ability to extract phenolic compounds from canola meal: water (WE) and various 80% organic solvent/water mixtures of methanol (ME), acetone (AE), ethanol (EE), butanol (BE), chloroform (CE) and hexane (HE). The in vitro antioxidant and anti-obesity properties of various extracts were investigated. Anti-obesity properties were studied using adipogenic differentiation inhibition of a murine mesenchymal stem cell line (C3H10T1/2) and a pancreatic lipase inhibition assay. AE, ME, and BE showed significant (p < 0.05) adipogenesis and pancreatic lipase inhibitory activities and may have more pharmacological properties. AE down-regulated the gene expression of the major adipogenic transcription factor, peroxisome proliferator-activated receptor gamma (PPARγ), correlating to phenolic content in a dose-dependent manner. The chemical characterization of AE revealed the presence of sinapic acid, ferulic acid, and kaempferol derivatives as main bioactive phenols.

White poplar: Targeted isolation of pancreatic lipase inhibitors

In order to discover new anti-obesity agents, twenty two natural extracts from medicinal plants and agricultural wastes were screened by pancreatic lipase inhibition assay. The methanolic extract of poplar wood (Populus alba L.), an important raw material for paper industry, showed promising inhibitory activity, its IC50 value was 7.15 μg/mL. Through bio-guided fractionation, the ethyl acetate fraction (E) was found to be the major contributor to the activity, IC50 = 4.7 μg/mL. Hence, it was subjected to bio-guided isolation and nine compounds (1–9) were identified from the active sub-fractions. The isolates were identified by means of one- and two-dimensional nuclear magnetic spectroscopy (1D, 2D-NMR), as well as high-performance liquid chromatography quadrupole time-of flight mass spectrometry (HPLC-QTOF-MS). (+)-Taxifolin (5) and (+)-ampelopsin (4) showed the highest activity, their IC50 values were 23.2 and 46.2 μM, respectively. The results indicated that flavonoids and phenolic acids in poplar wood could serve as potential candidates for development of new anti-obesity agents.

Saponin from purple eggplant (Solanum melongena L.) and their activity as pancreatic lipase inhibitor

Recently, obesity is one of the global problem, and orlistat is one of the known anti-obesity drugs, which works by inhibiting pancreatic lipase activity. Previous study has revealed that purple eggplant mesocarp, which contain alkaloids, flavonoids and saponins, has activity as pancreatic lipase inhibitor. On other hand, many class of saponin are inhibitor for the enzyme. Therefore, the aims of this study are to isolate and identify the saponins of purple eggplant (Solanum melongena) methanol extract and to test the activity as pancreatic lipase inhibitor. This study was conducted on four stages: 1) sample preparation, extraction and concentration of compounds from eggplant purple mesocarp; 2) saponin isolation by thin layer chromatography; 3) UV-Vis and FT-IR analysis to predict the class of the saponin isolates; and 4) pancreatic lipase inhibition assay. Extraction has been done by maceration using methanol as solvent, while the isolation was conducted by preparative thin layer chromatography using methanol: acetone (1:1) as eluent. Inhibition assay using titrimetric method with porcine pancreatic lipase (Sigma product) as enzyme, olive oil as substrate and orlistat as standard inhibitor. The result has showed that: the methanol extract of purple eggplant mesocarp contains two type of saponins that active as pancreatic lipase inhibitor, with the activity hundreds times higher than orlistat, at the same mass; and based on their UV-Vis and IR spectra, the isolates were predicted as steroid saponins.

Antiobesity and Antioxidant Potentials of Selected Palestinian Medicinal Plants.

We evaluated the antioxidant and porcine pancreatic lipase inhibition (PPLI) activities of 90 plants extracts. The antioxidant activity was measured using the free-radical scavenging capacity (DPPH) and reducing power (RP) assays. The pancreatic lipase inhibition assay was used to determine the PPLI activity of plant extracts. Among the 90 plant extracts examined, 41.0 % crude extracts showed antilipase activity of more than 50%. The most active plants by means of IC50 value were Camellia sinensis (0.5 mg/ml), Ceratonia siliqua (leaves) (0.8 mg/mL), Curcuma longa (0.8 mg/mL), Sarcopoterium spinosum (1.2 mg/mL), and Mentha spicata (1.2 mg/mL). The antioxidant activity of plant extracts using the DPPH and RP assays reveals comparable results. The most active antioxidant extracts using both assays were the leaves and fruit epicarp of Rhus coriaria, areal parts of Sarcopoterium spinosum, and leaves of Ceratonia siliqua. Our results suggest natural resources that possess strong antioxidant and pancreatic lipase inhibitory activities with potential applications in the treatment and prevention of obesity and overweight. The extracts of Camellia sinensis, Ceratonia siliqua, Curcuma longa, Sarcopoterium spinosum, and Mentha spicata were proved to have a great potential as antioxidants and antiobesity agents.

PHARMACOLOGICAL SCREENING OF ANTI-OBESITY POTENTIAL OF ACORUS CALAMUS LINN. IN HIGH FAT CAFETERIA DIET FED OBESE RATS

Objective: To study the anti-obesity potential of aqueous rhizome extract of Acoruscalamus Linn. (AREAC)in high fat diet fed obese rats.Methods: Adult strain male Wistar rats used in this study were fed with High Fat Diet (HFD) for 60 days. For the treatment groups,AREAC was administered in a dose levels of100, 200 and 300 mg/kgbw, orally once a day along with HFD. Rats fed with normal pellet chow were served as normal control. The effect of AREAC on physical parameterssuch as body weight, organ weight, fat pad weights and various biochemical parameterslike serum glucose, insulin, leptin,lipid profile, liver markers, kidney markers and oxidative stress markers were analysed.In-vitro pancreatic lipase inhibition assay of AREAC was also studied.Results: Data of in-vivo studies revealedsignificant (p&lt;0.05) reduction in percentage body weight gain, organ weights, fat pad weights and levels of serum glucose, insulin and leptin after treatment with AREAC in a dose dependent manner. Also, administration of AREAC significantly inhibited the increases in the concentrations of triglycerides, total cholesterol, LDL-cholesterol, VLDL-cholesterol, free-fatty acid and phospholipids in a dose dependent manner whereas, the level of HDL-cholesterol was found to be elevated on treatment. Moreover, on treatment with test drug,the elevated levels of serum liver and kidney markerssuch as AST, ALT, ALP, urea, creatinine were also brought back to near normalcy. Antioxidant status was found to be enhanced in liver tissues after treatment.In-vitro studies showed significant inhibition in the activity of pancreatic lipaseby AREAC.Conclusion: The data of the results obtained clearly depicted that AREAC was found to have pronounced anti-obesity activity particularly at the dose levels of 300 mg/kg bw.Key Words: Obesity, High Fat Diet, Leptin, AcoruscalamusLinn., Orlistat.

Bis-indole alkaloids from Tabernaemontana divaricata as potent pancreatic lipase inhibitors: molecular modelling studies and experimental validation

Obesity is the major leading cause of global mortality among metabolic disorders. Orlistat, a pancreatic lipase inhibitor, is the only approved drug of choice for the long term treatment of obesity. However, recent findings reported severe adverse effects with long term administration of orlistat. Plant-based natural products represent a vast reservoir of chemical entities that have the potential to treat various metabolic disorders. In the present study, we have performed a preliminary screening of local flora for pancreatic lipase inhibition assay, which highlighted the methanol extract of Tabernaemontana divaricata leaves (IC50 of 12.73 µg/mL). Molecular docking of the 38 alkaloids, reported from the leaves of T. divaricata, into the active site of pancreatic lipase led to the identification of furan bridged bis-indole alkaloids viz., Conophylline (1), Conophyllinine (2) and Conophyllidine (3) as potential leads, while Taberhanine (4), a monomeric indole alkaloid was found to exhibit comparatively poor docking score. Further, molecular docking analysis of the top three molecules (1–3) highlighted the importance of hydrophobic interactions of the dimeric extension with the lid domain of pancreatic lipase, which was not found with 4. Molecular dynamics simulations of 1–4 in complex with pancreatic lipase, further confirmed the docking results, wherein compound 4 was comparatively less stable (RMSD ≈ 1.5 A). Liquid chromatography–mass spectrometry analysis of the alkaloid-rich fraction of T. divaricata leaves indicated the presence of 1, while 2–4 were found to be absent. Molecule 1 was tested for pancreatic lipase inhibition potential, where it exhibited potent IC50 of 3.31 µM, comparable to that of orlistat (IC50 of 0.99 µM). Enzyme kinetic studies validated the in silico analyses, wherein compound 1 exhibited a competitive reversible inhibition of pancreatic lipase. The present study identified the bis-indole alkaloids of T. divaricata leaves as a new class of potent pancreatic lipase inhibitors.