Inhibition Of Β-glucosidase Research Articles

Rational Design and Synthesis of Novel Benzimidazole Derivatives as Potential β-Glucosidase Inhibitors

Background: β-Glucosidase has a variety of biological functions. A structural model for a potential β-glucosidase inhibitor has been proposed in the previous studies. Objective: A series f new diaryl derivatives linked through benzimidazole have been randomly and rationally designed, synthesized, and evaluated for their inhibitory activities against β-glucosidase (almond). The proposed structural model provides a new strategy for the design of potent β-glucosidase inhibitors. Methods: According to the model, a series of benzimidazole derivatives were designed and synthesized, and their inhibitory activity, Ki value, inhibitory type, and binding mode analysis (PDB ID: 2J7C) on β-glucosidase (almond) were evaluated. Results: Two compounds 7b and 7d were the best inhibitors with IC50 values of 7.86 M and 3.52 μM, respectively. Their Ki values were calculated to be 9.91 μM and 5.81 μM, respectively. Conclusion: The SAR analysis suggested that such benzimidazole derivatives might bind to the active site of β-glucosidase mainly through hydrogen bonds on o-trihydroxyphenol; the additional phenyl ring on the other side towards the solvent-exposed region played a very important role in improving their inhibitory activity against β-glucosidase

Design and synthesis of 6-C-alkyl-DMDP type nanomolar inhibitors of β-galactosidase and β-glucosidase based on broussonetine S and related derivatives

Broussonetine S (9), its C-1′ and C-10′ stereoisomers, and their corresponding enantiomers have been synthesized from enantiomeric arabinose-derived cyclic nitrones, with cross metathesis (CM), epoxidation and Keck asymmetric allylation as key steps. Glycosidase inhibition assays showed that broussonetine S (9) and its C-10′ epimer (10′-epi-9) were nanomolar inhibitors of bovine liver β-galactosidase and β-glucosidase; while their C-1′ stereoisomers were 10-fold less potent towards these enzymes. The glycosidase inhibition results and molecular docking calculations revealed the importance of the configurations of pyrrolidine core and C-1′ hydroxyl for inhibition potency and spectra. Together with the docking calculations we previously reported for α-1-C-alkyl-DAB derivatives, we designed and synthesized a series of 6-C-alkyl-DMDP derivatives with very simple alkyl chains. The inhibition potency of these derivatives was enhanced by increasing the length of the side chain, and maintained at nanomolar scale inhibitions of bovine liver β-glucosidase and β-galactosidase after the alkyl groups are longer than eight or ten carbons for the (6R)-C-alkyl-DMDP derivatives and their 6S epimers, respectively. Molecular docking calculations indicated that each series of 6-C-alkyl-DMDP derivatives resides in the same active site of β-glucosidase or β-galactosidase with basically similar binding conformations, and their C-6 long alkyl chains extend outwards along the hydrophobic groove with similar orientations. The increasing inhibitions of β-glucosidase and β-galactosidase with the number of carbon atoms in the side chains may be explained by improved adaptability of longer alkyl chains in the hydrophobic grooves. In addition, the lower β-glucosidase and β-galactosidase inhibitions of (6S)-C-alkyl-DMDP derivatives than their C-6 R stereoisomers can be attributed to the misfolding of their alkyl chains and resulted decreased adaptability in the hydrophobic groove. The work reported herein is valuable for design and development of more potent and selective inhibitors of β-galactosidase and β-glucosidase, which have potential in treatment of lysosomal storage diseases. Furthermore, part of the 6-C-alkyl-DMDP derivatives and their enantiomers were also tested as potential anti-cancer agents; all the compounds tested were found with moderate cytotoxic effects on MKN45 cells, which would indicate potential applications of these iminosugars in development of novel anticancer agents.

A remarkable change in inhibition potency and selectivity of isofagomine by simple N-modification

AbstractHerein, we present an alternative and elegant synthetic approach toward powerful β-glucosidase inhibitor isofagomine. Derivatizations of the ring nitrogen provided a selected set of N-modified isofagomine analogues. Biological evaluation of these compounds showed a remarkable change in potency as well as α/β-preference for various glycosidases from different sources when compared to the parent compound isofagomine. Overall, the conducted N-modification improved the potency against α-glucosidase from Saccharomyces cerevisiae (GH13). Coming along, significant diminished activities toward GH1 family β-glucosidases from three different sources have been observed for all tested derivatives. Moreover, and contrary to isofagomine, deactivations of β-galactosidase from Escherichia coli (GH2) as well as α-mannosidase from Canavalia ensiformis (GH38) have not been verified for this series of compounds. Graphical abstract

Conformational and Electronic Variations in 1,2- and 1,5a-Cyclophellitols and their Impact on Retaining α-Glucosidase Inhibition.

Glycoside hydrolases (glycosidases) take part in myriad biological processes and are important therapeutic targets. Competitive and mechanism-based inhibitors are useful tools to dissect their biological role and comprise a good starting point for drug discovery. The natural product, cyclophellitol, a mechanism-based, covalent and irreversible retaining β-glucosidase inhibitor has inspired the design of diverse α- and β-glycosidase inhibitor and activity-based probe scaffolds. Here, we sought to deepen our understanding of the structural and functional requirements of cyclophellitol-type compounds for effective human α-glucosidase inhibition. We synthesized a comprehensive set of α-configured 1,2- and 1,5a-cyclophellitol analogues bearing a variety of electrophilic traps. The inhibitory potency of these compounds was assessed towards both lysosomal and ER retaining α-glucosidases. These studies revealed the 1,5a-cyclophellitols to be the most potent retaining α-glucosidase inhibitors, with the nature of the electrophile determining inhibitory mode of action (covalent or non-covalent). DFT calculations support the ability of the 1,5a-cyclophellitols, but not the 1,2-congeners, to adopt conformations that mimic either the Michaelis complex or transition state of α-glucosidases.

Production, purification, and determination of the biochemical properties of β-glucosidase in Trichoderma koningii via solid substrate fermentation.

The β-glucosidase enzyme was obtained from Trichoderma koningii Oudem. NRRL 54330 under optimal conditions by solid substrate fermentation (SSF) using corn cobs as substrate. The enzyme was purified by two-step procedures, ammonium sulphate precipitation and cefarose-4B-l-tyrosine-1-naphthylamine hydrophobic interaction chromatography, followed by biochemical and kinetic characterisation. The β-glucosidase was obtained from T.koningii using ground corn cob as substrate and Na2HPO4, pH 9, as humidification medium. The optimum conditions for enzyme production by SSF were 30 °C and 6days. The purification efficiency of the obtained β-glucosidase was calculated to be 22.56-fold with a yield of 73.51 %. In the determination of β-glucosidase activity, p-nitrophenyl-β-d-glucopyranoside (pNPG) substrate was used, and the optimum pH and temperature values at which β-glucosidase showed high activity were determined to be pH 3.0 and 75 °C. The purity of the enzyme and the presence/number of subunits were checked using two different electrophoretic methods, SDS-PAGE and NATIVE-PAGE electrophoretic methods. The K m and V max values of the purified enzyme were determined to be 0.16 mM and 2000 EU respectively. Itwas also found that d-(+)-glucose and δ-gluconolactone inhibitors exhibited competitive inhibition of β-glucosidase in the presence of pNPG.

Genome-based approach to evaluate the metabolic potentials and exopolysaccharides production of Bacillus paralicheniformis CamBx3 isolated from a Chilean hot spring.

In the present study, a thermophilic strain designated CamBx3 was isolated from the Campanario hot spring, Chile. Based on 16S rRNA gene sequence, phylogenomic, and average nucleotide identity analysis the strain CamBx3 was identified as Bacillus paralicheniformis. Genome analysis of B. paralicheniformis CamBx3 revealed the presence of genes related to heat tolerance, exopolysaccharides (EPS), dissimilatory nitrate reduction, and assimilatory sulfate reduction. The pangenome analysis of strain CamBx3 with eight Bacillus spp. resulted in 26,562 gene clusters, 7,002 shell genes, and 19,484 cloud genes. The EPS produced by B. paralicheniformis CamBx3 was extracted, partially purified, and evaluated for its functional activities. B. paralicheniformis CamBx3 EPS with concentration 5 mg mL-1 showed an optimum 92 mM ferrous equivalent FRAP activity, while the same concentration showed a maximum 91% of Fe2+ chelating activity. B. paralicheniformis CamBx3 EPS (0.2 mg mL-1) demonstrated β-glucosidase inhibition. The EPS formed a viscoelastic gel at 45°C with a maximum instantaneous viscosity of 315 Pa.s at acidic pH 5. The present study suggests that B. paralicheniformis CamBx3 could be a valuable resource for biopolymers and bioactive molecules for industrial applications.

N-2 Alkylated analogues of aza-galactofagomine as potential inhibitors of β-glucosidase

The synthesis of four N-2-alkylated aza-galactofagomine (AGF) analogues was achieved by intermolecular reductive hydrazination or alkylation of suitably protected AGF. The synthesized compounds were evaluated as potential ?-glucosidase inhibitors. The preliminary screening of inhibitor activity, conducted with sweet almond ?-glucosidase immobilized in agar, as well as the standard inhibition assay with the same enzyme, showed the inhibitory potency of the synthesized analogues. In addition, these results are in a good agreement with the docking analysis of the human acid ?-glucosidase, the enzyme implicated in Gaucher?s disease.

Effect-Directed Assays and Biological Detection Approaches Coupled with Thin-Layer Chromatography as an Evolving Hyphenated Technique: A Comprehensive Review.

Bioautography is a technique for the detection of biological activity that combines the elements of planar chromatography. Its hyphenated variants are widely used in the screening of natural products possessing biological activity. It can be used in the activity-based screening of phytochemical ingredients by employing various enzyme processes and reactions and facilitates the rapid determination of bioactive compounds in pant samples. To give a comprehensive overview of effect-directed assays and biological detection approaches used in conjugation with thin layer chromatography technique. The present review article attempts to throw light on the various aspects of bioautography, including its types and applications, thereby giving its concise overview and its relevance in the field of natural product screening. Various search engines were used for the literature survey, including Google Scholar, Semantic Scholar, PubMed, ResearchGate and Scopus. Bioautography has wide-ranging uses in the screening of compounds such as antioxidants, antifungals, antimicrobials, estrogenic, antitumors, and various enzyme inhibitors compounds like α and β-glucosidase inhibitors and α-amylase inhibitors. Bioautography serves to be an effective tool for the isolation of bioactive phytochemicals, thereby allowing us to scientifically validate the biological activities of various compounds, which can then be utilized for making potent medications for various diseases.

Elucidating the Ionic Liquid-Induced Mixed Inhibition of GH1 β-Glucosidase H0HC94.

Deciphering the ionic liquid (IL) tolerance of glycoside hydrolases (GHs) to improve their hydrolysis efficiency for fermentable sugar synthesis in the "one-pot" process has long been a hurdle for researchers. In this work, we employed experimental and theoretical approaches to investigate the 1-ethyl-3-methylimidazolium acetate ([C2C1im][MeCO2])-induced inhibition of GH1 β-glucosidase (H0HC94) from Agrobacterium tumefaciens 5A. At 10-15% [C2C1im][MeCO2] concentration, H0HC94 experiences competitive inhibition (R2 = 0.97, alpha = 2.8). As the IL content increased to 20-25%, the inhibition pattern shifted to mixed-type inhibition (R2 = 0.98, alpha = 3.4). These findings were further confirmed through characteristic inhibition plots using Lineweaver-Burk plots. Atomistic molecular dynamics simulations conducted with 0% [C2C1im][MeCO2], 10% [C2C1im][MeCO2], and 25% [C2C1im][MeCO2] revealed the accumulation of [C2C1im]+ at the negatively charged active site of H0HC94 in 10% [C2C1im][MeCO2], supporting the occurrence of competitive inhibition at lower IL concentrations. At higher IL concentrations, the cations and anions bound to the secondary binding sites (SBSs) of H0HC94, leading to a tertiary conformational change, as captured by the principal component analysis based on the free-energy landscape and protein structure networks. The altered conformation of H0HC94 affected the interaction with [C2C1im][MeCO2], which could possibly shift the inhibition from competitive to more mixed-type (competitive + noncompetitive) inhibition, as observed in the experiments. For the first time, we report a combined experimental and theoretical insight behind the mixed inhibition of a GH1 β-glucosidase. Our findings indicated the role of SBS in IL-induced inhibition, which could aid in developing more IL-tolerant β-glucosidases for biorefinery applications.

Bioactivity-Guided Synthesis: In Silico and In Vitro Studies of β-Glucosidase Inhibitors to Cope with Hepatic Cytotoxicity.

The major cause of hyperglycemia can generally be attributed to β-glucosidase as per its involvement in non-alcoholic fatty liver disease. This clinical condition leads to liver carcinoma (HepG2 cancer). The phthalimides and phthalamic acid classes possess inhibitory potential against glucosidase, forming the basis for designing new phthalimide and phthalamic acid analogs to test their ability as potent inhibitors of β-glucosidase. The study also covers in silico (molecular docking and MD simulations) and in vitro (β-glucosidase and HepG2 cancer cell line assays) analyses. The phthalimide and phthalamic acid derivatives were synthesized, followed by spectroscopic characterization. The mechanistic complexities associated with β-glucosidase inhibition were identified via the docking of the synthesized compounds inside the active site of the protein, and the results were analyzed in terms of the best binding energy and appropriate docking pose. The top-ranked compounds were subjected to extensive MD simulation studies to understand the mode of interaction of the synthesized compounds and binding energies, as well as the contribution of individual residues towards binding affinities. Lower RMSD/RMSF values were observed for 2c and 3c, respectively, in the active site, confirming more stabilized, ligand-bound complexes when compared to the free state. An anisotropic network model was used to unravel the role of loop fluctuation in the context of ligand binding and the dynamics that are distinct to the bound and free states, supported by a 3D surface plot. An in vitro study revealed that 1c (IC50 = 1.26 µM) is far better than standard acarbose (2.15 µM), confirming the potential of this compound against the target protein. Given the appreciable potential of the candidate compounds against β-glucosidase, the synthesized compounds were further tested for their cytotoxic activity against hepatic carcinoma on HepG2 cancer cell lines. The cytotoxicity profile of the synthesized compounds was performed against HepG2 cancer cell lines. The resultant IC50 value (0.048 µM) for 3c is better than the standard (thalidomide: IC50 0.053 µM). The results promise the hypothesis that the synthesized compounds might become potential drug candidates, given the fact that the β-glucosidase inhibition of 1c is 40% better than the standard, whereas compound 3c holds more anti-tumor activity (greater than 9%) against the HepG2 cell line than the known drug.

Expression, activity, and consequences of biochemical inhibition of α- and β-glucosidases in different life stages of Culex quinquefasciatus.

Mosquitoes have a wide range of digestive enzymes that enable them to utilize requisite blood and sugar meals for survival and reproduction. Sugar meals, typically derived from plant sources, are critical to maintain energy in both male and female mosquitoes, whereas blood meals are taken only by females to complete oogenesis. Enzymes involved in sugar digestion have been the subject of study for decades but have been limited to a relatively narrow range of mosquito species. The southern house mosquito, Culex quinquefasciatus, is of public health importance and seldom considered in these types of studies outside of topics related to Bacillus sphaericus, a biocontrol agent that requires interaction with a specific gut-associated α-glucosidase. Here we sought to describe the nature of α-glucosidases and unexplored β-glucosidases that may aid Cx. quinquefasciatus larvae in acquiring nutrients from cellulosic sources in their aquatic habitats. Consistent with our hypothesis, we found both α- and β-glucosidase activity in larvae. Interestingly, β-glucosidase activity all but disappeared at the pupal stage and remained low in adults, while α-glucosidase activity remained in the pupal stage and then exceeded larval activity by approximately 1.5-fold. The expression patterns of the putative α- and β-glucosidase genes chosen did not consistently align with observed enzyme activities. When the α-glucosidase inhibitor acarbose was administered to adults, mortality was seen especially in males but also in females after two days of exposure and key energetic storage molecules, glycogen and lipids, were significantly lower than controls. In contrast, administering the β-glucosidase inhibitor conduritol β-epoxide to larvae did not produce mortality even at the highest soluble concentration. Here we provide insights into the importance of α- and β-glucosidases on the survival of Cx. quinquefasciatus in their three mobile life stages.

Development of nanofunctionalized oxovanadium(IV) complex and its anticancer, antidiabetic, DNA cleavage and cell imaging studies

VO(IV) complex is little toxic and highly effective than vanadium salts. A vanadyl metal complex from 8-formyl-7-hydroxy-4-methyl coumarin derivative has been synthesized and functionalized with copper nanoparticles. The Spectrochemical studies such as UV, FTIR, 1NMR and ESR spectra were recorded to characterize the ligand(CUAP), Vanadyl complex[VO(CUAP)SO4] and nano Cu-VO(IV)complex efficiently. The structural studies of vanadyl complex confirmed that the ligand coordinate with metal through nitrogen atom of azomethine, carbonyl oxygen and phenolic oxygen. ESR spectrum of vanadyl complex revealed the covalent nature. XRD pattern of nano Cu-VO(IV) complex indicated the crystalline nature and the average particle size was 20.91 nm. SEM image of nano Cu-VO(IV) complex showed that the nano particles accumulated to form spherical shaped particles. The particle size obtained from Transmission Electron Microscopy of nano functionalized metal complex is ∼ 20 nm. It is closely matched to the particle size calculated from XRD results. Fluorescence of vanadyl complex and nano Cu-VO(IV) complex exhibit the emission from 270 to 900 nm range with significant fluorescence at ∼ 750 nm. The DNA cleavage of all the compounds was evaluated using Agarose gel electrophoresis technique and showed greater cleavage of vanadyl complex. The anticancer activity of compounds was carried out against two cancer cell lines viz Human Breast Cancer Cell line (MCF-7) and Human Leukemia Cancer Cell Line(K-562). Oxovanadium complex exhibited good anticancer activities than ligand and nano-functionalized complex. The antidiabetic activities of vanadyl and nano functionalized complexes were studied against α-Amylase and β-Glucosidase inhibition assay. In this study vanadyl complex showed higher inhibition activity on α-Amylase compared with standard Acarbose. The bioimaging of nano-functionalized metal complex showed high fluorescent properties. The molecular docking study of ligand and vanadyl complex showed greater docking results with CDK2 receptor.

The serine/threonine protein kinase VSKM1 promotes cold stress resistance of the postharvest Volvariella volvacea by enhancing mitoribosome activity

The edible straw mushroom Volvariella volvacea has a desirable taste and diverse effects on health; however, it undergoes rapid autolysis at 4 °C. Spray treatment with the β-glucosidase inhibitor DGlucono-1,5-lactone (DGase) at 100 µmol/L maintained the quality of V. volvacea during storage at 4 °C for 72 h. Enhanced alkaline protease and neutral protease activities can act as indicators of cold stress resistance (CSR) in V. volvacea, as well as the stable ATP level. The total sugar and amino acid content is negatively correlated with CSR in V. volvacea. DGase treatment enhanced the CSR of V. volvacea by inducing the overexpression of genes in the mitochondrial ribosome (mitoribosome). Overexpression of a serine/threonine-protein kinase gene in V. volvacea (VSKM1), which promotes mitoribosome activity in yeast, enhanced freezing tolerance. Our findings indicated that overexpression of VSKM1 promoted CSR by enhancing mitoribosome activity for the quality retention of postharvest V. volvacea.

GBA inhibition suppresses ovarian cancer growth, survival and receptor tyrosine kinase AXL-mediated signaling pathways.

The poor outcome of advanced ovarian cancer under conventional therapy necessitates new strategies to improve therapeutic efficacy. β-glucosidase (encoded by GBA) is a lysosomal enzyme and is involved in sphingolipids metabolism. Recent studies revealed that β-glucosidase plays a role in cancer development and chemoresistance. In this work, we systematically evaluated the expression and role of GBA in ovarian cancer. Our work demonstrates that inhibition of β-glucosidase has therapeutic potential for ovarian cancer. Gene Expression Profiling Interactive Analysis database, western blot and immunohistochemistry analyses of patient samples demonstrated that GBA mRNA and protein expression levels were significantly increased in ovarian cancer compared to normal tissues. Functional studies using gain-of- function and loss-of-function approaches demonstrated that GBA overexpression did not affect growth and migration but alleviated cisplatin's efficacy in ovarian cancer cells. In addition, GBA depletion resulted in growth inhibition, apoptosis induction, and enhancement of cisplatin's efficacy. Of note, we found that GBA inhibition specifically decreased receptor tyrosine kinase AXL level, leading to the suppression of AXL-mediated signaling pathways. Our data suggest that GBA represents a promising target to inhibit AXL signaling and overcome cisplatin resistance in ovarian cancer.

Curcumin nanoparticles: physicochemical fabrication, characterization, antioxidant, enzyme inhibition, molecular docking and simulation studies.

Curcumin is an extensively studied natural compound due to its extensive biological applications. However, there are some drawbacks linked to this compound such as poor absorption, low water-solubility, quick systemic elimination, fast metabolism, poor pharmacokinetics, low bioavailability, low penetration targeting efficacy and low stability. To overcome these drawbacks, curcumin is encapsulated in nano-carriers. In the current studies, we synthesized nanoparticles of curcumin without using nanocarriers by different methods such as nano-suspension (Cur-NSM), sonication (Cur-SM) and anti-solvent precipitation (Cur-ASP) to enhance the solubility of curcumin in water. The prepared nanoparticles were characterized by FTIR, SEM and XRD analysis. These curcumin nanoparticles were screened for their solubilities in water, DPPH scavenging, amylase, α-glucosidase and β-glucosidase enzymatic activities. The particle size of nano-curcumin was found to be in the 47.4-98.7 nm range. The reduction in particle size of curcumin dramatically increases its solubility in water to 79.2 μg mL-1, whereas the solubility of curcumin is just 0.98 μg mL-1. Cur-ASP showed the highest free radical scavenging potential (48.84 ± 0.98%) which was comparable with standard BHT (50.48 ± 1.11%) at 75.0 μg mL-1. As well, Cur-ASP showed the highest inhibition of α-amylase (68.67 ± 1.02%), α-glucosidase (58.30 ± 0.52%), and β-glucosidase (64.80 ± 0.43%) at 100 μg mL-1 which is comparable with standard drug acarbose. The greater surface area of nanoparticles exposes the various groups of curcumin for blocking the binding sites of enzymes. This strategy may be helpful in designing curcumin as a potent therapeutic agent against diabetes mellitus. Further, the molecular interactions of curcumin with α-amylase, α-glucosidase, β-glucosidase, and polyphenol oxidase were assessed by analyzing the plausible binding modes of curcumin in the binding pocket of each receptor. The best binding mode of curcumin was used to make complexes with the target proteins and their stability was confirmed by 50 ns MD simulation.

Phyto-fabrication of ultrafine nanoscale holmium oxide HT-Ho2O3 NPs and their biomedical potential.

In this study holmium oxide nanoparticles (Ho2O3 NPs) are fabricated using Hyphaene thebaica extracts as a bioreductant. The XRD pattern of HT-Ho2O3 NPs (product from phyto-reduction) suggested that the nanoparticles are crystalline with no impurities. Scherrer approximation revealed grain sizes of ∼10 nm. The HR-TEM revealed HT-Ho2O3 NPs possessed a quasi-spherical morphology complemented by SEM and the particle sizes were in the range of 6-12 nm. The infrared spectra revealed characteristic Ho-O bonding at ∼603 cm-1. Raman spectra indicated five main peaks positioned at 156 cm-1, 214 cm-1, 328 cm-1, 379 cm-1 and 607 cm-1. Eg (optical bandgap) was found to be 5.1 eV. PL spectra indicated two major peaks at 415 nm and 607 nm. EDS spectra confirmed the elemental presence of holmium (Ho). Spotty rings were obtained during the SAED measurement which indicated crystallinity of HT-Ho2O3 NPs. The HT-Ho2O3 NPs were further analyzed for their antioxidant, anti-angiogenic and cytotoxic properties. The antioxidant potential was moderate i.e., 43.40 ± 0.96% at 1000 μg mL-1 which decreased in a dose dependent manner. Brine shrimp lethality was highest at 1000 μg mL-1 with the LC50 320.4 μg mL-1. Moderate anti-angiogenic potential was observed using in ova CAM assay. MTT bioassay revealed that the HT-Ho2O3 NPs inhibited the 3T3 cells (IC50 67.9 μg mL-1), however, no significant inhibition was observed against MCF-7 cells. α-Amylase and β-glucosidase inhibition revealed that the HT-Ho2O3 NPs can be of use in controlling blood glucose levels. Overall, it can be concluded that biosynthesis using aqueous extracts can be a suitable alternative in finding ecofriendly paradigms for the synthesis of nanoparticles. We suggest extended research into the bioreduced Ho2O3 NPs for establishing their biomedical potential and toxicity.

Citrullus lanatus-encased zinc oxide nanoparticles as potential anti-diabetic, anti-inflammatory and antibacterial agents: A new strategy towards biocompatible nano-drugs

This research aimed to develop a rapid, cheap, and environmentally friendly method for synthesizing zinc oxide nanoparticles (ZnONPs) from Citrullus lanatus (Cl) seed extract and the Cl-ZnONPs were analyzed using several techniques to determine their properties. These included UV–vis spectroscopy, XRD, FTIR, SEM, and TEM. Cl-ZnONPs have sizes between 10 and 50 nm and are spherical in form. Compared to Cl seed extract, Cl-ZnONPs displayed significantly higher levels of antioxidant activity. Comparatively to α-amylase, Cl-ZnONPs showed much stronger inhibition of β-glucosidase and DPPIV (anti-diabetic activity). Cl-ZnONPs and Cl-seed extract were tested for their ability to reduce inflammation, demonstrating potent in vitro anti-inflammatory properties (membrane stabilization, proteinase inhibitory, and lipoxygenase). In addition, the Cl-ZnONPs' antibacterial activity revealed that they had a significantly greater effect on inhibiting the growth of Gram-negative bacteria such as Escherichia coli and Pseudomonas aeruginosa. Synthesized Cl- ZnONPs are promising solutions for treating inflammatory and diabetic concerns, as suggested by the outcomes. In conclusion, the biosynthesized Cl-ZnONPs shown in this study provide a synthetic substance replacement that is safe and effective in various biomedical and pharmaceutical applications, including those requiring antioxidant, antibacterial, and anti-inflammatory properties.

Zearalenone-14-Glucoside Is Hydrolyzed to Zearalenone by β-Glucosidase in Extracellular Matrix to Exert Intracellular Toxicity in KGN Cells.

As one of the most important conjugated mycotoxins, zearalenone-14-glucoside (Z14G) has received widespread attention from researchers. Although the metabolism of Z14G in animals has been extensively studied, the intracellular toxicity and metabolic process of Z14G are not fully elucidated. In this study, the cytotoxicity of Z14G to human ovarian granulosa cells (KGN) and the metabolism of Z14G in KGN cells were determined. Furthermore, the experiments of co-administration of β-glucosidase and pre-administered β-glucosidase inhibitor (Conduritol B epoxide, CBE) were used to clarify the mechanism of Z14G toxicity release. Finally, the human colon adenocarcinoma cell (Caco-2) metabolism model was used to verify the toxicity release mechanism of Z14G. The results showed that the IC50 of Z14G for KGN cells was 420 μM, and the relative hydrolysis rate of Z14G on ZEN was 35% (25% extracellular and 10% intracellular in KGN cells). The results indicated that Z14G cannot enter cells, and Z14G is only hydrolyzed extracellularly to its prototype zearalenone (ZEN) by β-glucosidase which can exert toxic effects in cells. In conclusion, this study demonstrated the cytotoxicity of Z14G and clarified the toxicity release mechanism of Z14G. Different from previous findings, our results showed that Z14G cannot enter cells but exerts cytotoxicity through deglycosylation. This study promotes the formulation of a risk assessment and legislation limit for ZEN and its metabolites.

Rottlerin Stimulates Exosome/Microvesicle Release Via the Increase of Ceramide Levels Mediated by Ampk in an In Vitro Model of Intracellular Lipid Accumulation

Exosomes/microvesicles originate from multivesicular bodies that allow the secretion of endolysosome components out of the cell. In the present work, we investigated the effects of rottlerin, a polyphenol, on exosome/microvesicle secretion in a model of intracellular lipid trafficking impairment, and elucidated the mechanism of action. In a model of lipid trafficking impairment in C6 glia cells, rottlerin increased ceramide levels, while decreasing hexosylceramide content. This was accompanied by increased exosome/microvesicle secretion, thereby reducing the concentration of lipids in the endolysosomal compartment. The reduction of hexosylceramide levels by rottlerin was attributed to the increase of β-glucosidase (glucosylceramidase) activity, and the effects of rottlerin were abrogated by β-glucosidase inhibitors such as isofagomine D-tartrate and AMP-deoxynojirimycin. Moreover, treatment with ML-266, a potent activator of the β-glucosidase enzyme, recapitulated the effects of rottlerin on the sphingolipid profile and exosome/microvesicle secretion. Finally, inhibition of AMPK (AMP-activated protein kinase) using compound C prevented both exosome/microvesicle secretion and the elimination of endolysosome lipids, which were promoted by rottlerin. The results showed that the decrease in intracellular lipid deposition induced by rottlerin was mediated by β-glucosidase activation and exosome/microvesicle release via the AMPK pathway. Rottlerin consumption could represent an additional health benefit in lysosomal deposition diseases.

Phytochemical analysis and inhibitory effects of Calligonum polygonoides on pancreatic α-amylase and β-glucosidase enzymes.

To estimate the existence of phyto-chemicals and then to determine the antidiabetic activity against α-amylase and β-glucosidase inhibition . The study was carried out by following standard procedures. Phytochemicals analysis indicated the presence of different phytochemicals. The total phenolic content was 6.055 mg GAE/g and the total flavonoid content was 5.706 mg RU/g in the plant extract. The total saponins, alkaloids, and tannins contents were (0.044%), (2.88%) and (2.862 nm) respectively. α-amylase inhibition activity of Calligonum polygonoides (CP) extract was 70% with IC50 of 610 μg/mL and that of β-gluco-sidase inhibition activity was 65% with IC50 of 640 µg/mL. The findings reported for the first time the antidiabetes-promoting effects of an extract of CP, thus validating their promising anti-diabetes potential.