Acetylcholinesterase Inhibition Assay Research Articles

In vitro anticancer, antioxidant, antimicrobial, antileishmanial, enzymes inhibition and in vivo anti-inflammatory activities of organotin(IV) derivatives of 4-bromophenoxyacetic acid

Organotin(IV) derivatives {(n-C4H9)3SnL (1), (CH3)3SnL (2), (n-C4H9)2SnL2 (3) and (CH3)2SnL2 (4)} of 4-bromophenoxyacetic acid (HL) were synthesized and characterized by elemental, FT-IR, NMR and single crystal XRD diffraction analyses. The carboxylate ligand showed bridging/chelating bidentate coordination. The complexes 1 and 2 have adopted polymeric chain structures consisting tin atoms in distorted trigonal bipyramidal geometry. Complex 3 has shown highest DPPH radical scavenging (IC50 = 33.46 µg/mL) and antileishmanial activity (IC50 = 27.06 µg/mL). Complex 3 was also the most efficient agent in acetylcholinesterase (IC50 = 5.12 µg/mL), butyrylcholinesterase (IC50 = 13.79 µg/mL) and Monoamine oxidase (IC50 = 8.50 µg/mL) inhibition assays. Complex 4 (IC50 = 18.73 µg/mL) was the most potent ABTS radical scavenger. Complex 1 with IC50 values of 27.50 and 71.32 µg/mL was the most efficient α-glucosidase and dipeptidyl peptidase-4 enzyme inhibitor, respectively. While complex 2 was found the most potent Cyclooxygenase-2 (7.81 µg/mL) and 5-Lioxygenase (6.69 µg/mL) inhibitor. MTT assay revealed highest inhibitory potency of complexes 4 (IC50 = 8.035 ± 0.05 μg/ml), and 1 (IC50 = 10.16±0.17μg/ml) towards the brain cancer cell line. The in vivo study performed on mice have shown potent anti-inflammatory effects of 1 and 2 comparable to the standard, indomethacin.

Newly synthesized Bis-Hydrazono[1,3,4] thiadiazoles as Anti-Alzheimer’s Agents: Greener past and In-Vitro acetylcholinesterase inhibition assay investigations

The designed and synthesized multifunctional properties of bis-hydrazono[1,3,4]thiadiazoles became important for developing an efficient medication for Alzheimer’s disease. Thiadiazoles were characterized and studied towards this target as an acetylcholinesterase inhibitor (ChEIs). In this study, one-pot synthetic strategy was applied for the synthesis of bis-hydrazono[1,3,4]thiadiazole series from 2,2′-(1,2-diphenylethane-1,2-diylidene)bis(hydrazine-1-carbodithioic acid) and hydrazonoyl chlorides. The greener pastures, in silico, an environmentally-friendly and free computer-aided method, bioactivity studies of the bis-hydrazono[1,3,4]thiadiazoles 4a-h exhibited various possible interesting inhibitory activities against AChE showing similar behaviour to the approved drugs, Donepezil, Galantamine and Rivastigmine. For a fair comparison, the superpositions of 4a-h, and Donepezil, Galantamine and Rivastigmine docked together into the functional domain (binding pocket) of 1EVE to reveal interesting different molecular laydowns of the compounds. On the other side, the display of the molecules before and after docking was discussed. The binding affinity slightly differs between Donepezil, Galantamine, Rivastigmine, and the ligands 4a-h. The range of the recorded binding affinity for 4a-h is from (−9.4 to −8.4 kcal/mol−1), whereas the binding affinity for Donepezil, Galantamine and Rivastigmine is (−11.1, −9.8 and −8.0 kcal/mol−1 respectively), which is higher than all the prepared ligands.Furthermore, the binding amino acids also varied among the studied compounds in this study. Phe290, Phe330, Phe331, Trp279, Tyr70, Tyr121 and Tyr334 are the common amino acids binding with the FDA-approved AChE inhibitors for treating AD, Donepezil, Galantamine and Rivastigmine and ligands. Notably, the number of hydrophobic and hydrogen interactions studied between the ligands 4a-h and the Donepezil, Galantamine and Rivastigmine drugs were compared as a preliminary indicator towards a successful inhibitor. The comparative study in this research work aims to rank our compounds with respect to the approved medications. This is a friendly environmental preliminary helpful indication before leaping into time and energy-consuming experimental work. Organ toxicological endpoints and toxicity-predicted activity were obtained using the ProTox-II web server for both the approved medication and ligands. Later, in-vitro acetylcholinesterase inhibition assays were conducted to assess the efficacy of bis-hydrazono[1,3,4]thiadiazoles 4a-h as inhibitors of acetylcholinesterase (AChE) in comparison to Donepezil. Results indicated promising inhibition activities for example 4 h, 4 g and 4d to the testing drug Donepezil for breaking down the neurotransmitter acetylcholine.

Suitability of human HepaRG cells and liver spheroids as in vitro model to investigate the bioactivation of the organothiophosphate pesticide parathion

Organophosphorus compounds (OP) constitute a large group of chemicals including pesticides and nerve agents. Organothiophosphate pesticides require cytochrome P450-mediated oxidative desulphuration in the liver to form corresponding oxons, which are potent inhibitors of the enzyme acetylcholinesterase (AChE). Human HepaRG cells are a promising tool to study liver-specific functions and have been shown to maintain drug metabolizing enzymes. This research describes for the first time the in vitro metabolic activation of an organothiophosphate to its active oxon by two different HepaRG cell-based models. Monolayer cultures and liver spheroids were exposed to the model OP parathion and the quantification of the corresponding oxon was performed with an AChE inhibition assay. Our results showed a time- and dose-dependent cytochrome P450 catalyzed bioactivation and a superior metabolism capacity of the monolayer HepaRG model in comparison with the liver spheroids. Finally, HepaRG cells can be assessed as a metabolically competent cell model intermediate between cell-free preparations and intact animals and as suitable to study OP metabolism in the human liver.

New amide and diterpene alkaloids with anticholinesterase activity from Delphinium cyphoplectrum roots.

The cholinergic hypothesis posits a robust correlation between the onset of Alzheimer's disease and a pronounced deficit in acetylcholine, a pivotal neurotransmitter crucial for the central cholinergic nervous system's function, pivotal for memory and learning. Diterpene alkaloids exhibit intricate and distinctive chemical structures that facilitate their passage through the blood-brain barrier. Moreover, their potent pharmacological attributes render them promising candidates for addressing central nervous system disorders. This investigation aims to scrutinize the alkaloidal composition of Delphinium cyphoplectrum (Ranunculaceae) roots, further exploring their anticholinesterase inhibitory activity and mode of inhibition. Innovative chromatography techniques were repetitively employed to purify the alkaloids. Acetylcholinesterase (AChE) inhibition assays were conducted using Ellman's tests. The mode of inhibition was meticulously characterized through Michaelis-Menten, and Lineweaver-Burk plots. Conducting molecular docking studies, we employed the AUTO DOCK 4.2 software package. Eight alkaloids were identified including five C19-diterpene alkaloids (6,14,16,18-tetramethoxy-1,7,8-trihydroxy-4-methylaconitane (1), 6,16,18-trimethoxy-1,7,8,14-tetrahydroxy-4-methylaconitane (2), 6,8,16,18-tetramethoxy-1,7,14-trihydroxy-4-methylaconitane (3), 6,14,16-trimethoxy-1,7,8,18-tetrahydroxy-4-methylaconitane (4), and 14-O-acetyl-8,16-dimethoxy-1,6,7,18-tetrahydroxy-4-methylaconitane (5)), an epoxy C18-diterpene alkaloid (6,8,16-trimethoxy-1,7,14-trihydroxy-3,4-epoxyaconitane (6)), a known (pyrrolidin-2-one (7) and an undescribed amide alkaloid (1-(2'-hydroxylethylamine)-3,5,5,-trimethyl-1,5-dihydro-2H-pyrrol-2-one (8). All diterpene alkaloids underwent assessment for acetylcholinesterase (AChE) inhibition assay and displayed noteworthy AChE activity, surpassing that of the reference drug (with IC50 values of 13.7, 21.8, 23.4, 28.2, 40.4, and 23.9 for compounds 1-6, respectively, in comparison to 98.4 for Rivastigmine). Analysis of Michaelis-Menten and Lineweaver-Burk plots represents an uncompetitive mode of inhibition for compound 1 on AChE. Notably, computational docking simulations indicated that all diterpene alkaloids were accommodated within the same enzymatic cleft as the reference ligand, and displaying superior free binding energy values (from - 10.32 to -8.59 Kcal.mol-1) in contrast to Rivastigmine (-6.31 Kcal.mol-1). The phytochemical analysis conducted on the roots of Delphinium cyphoplectrum yielded the identification of eight alkaloidal compounds including one C18-diterpene, five C19-diterpene, one pyrrolidine and one amide alkaloids. AChE inhibition assay and molecular simulations unveiled remarkable significant potency attributed to the C19-diterpene alkaloids by the order of 1 > 2 > 3,6 > 4 > 5. Presence of hydroxyl group on C-1, C-7, C-8, C-14, and C-18 increased the effect. The best in vitro activity was recorded for compound 1 able to bind to Asp72 in the narrow region of PAS, while interacting by pi-sigma with Phe330 at the hydrophobic region of the gorge involving the acyl and choline binding site. This observation underscores the substantial promise of this category of natural products in the realm of drug discovery for Alzheimer's Disease, offering a compelling avenue for further research and therapeutic development.

African Under-Utilized Medicinal Leafy Vegetables Studied by Microtiter Plate Assays and High-Performance Thin-Layer Chromatography-Planar Assays.

Biological activities of six under-utilized medicinal leafy vegetable plants indigenous to Africa, i.e., Basella alba, Crassocephalum rubens, Gnetum africanum, Launaea taraxacifolia, Solanecio biafrae, and Solanum macrocarpon, were investigated via two independent techniques. The total phenolic content (TPC) was determined, and six microtiter plate assays were applied after extraction and fractionation. Three were antioxidant in vitro assays, i.e., ferric reducing antioxidant power (FRAP), cupric reduction antioxidant capacity (CUPRAC), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging, and the others were enzyme (acetylcholinesterase, butyrylcholinesterase, and tyrosinase) inhibition assays. The highest TPC and antioxidant activity from all the methods were obtained from polar and medium polar fractions of C. rubens, S. biafrae, and S. macrocarpon. The highest acetyl- and butyrylcholinesterase inhibition was exhibited by polar fractions of S. biafrae, C. rubens, and L. taraxacifolia, the latter comparable to galantamine. The highest tyrosinase inhibition was observed in the n-butanol fraction of C. rubens and ethyl acetate fraction of S. biafrae. In vitro assay results of the different extracts and fractions were mostly in agreement with the bioactivity profiling via high-performance thin-layer chromatography-multi-imaging-effect-directed analysis, exploiting nine different planar assays. Several separated compounds of the plant extracts showed antioxidant, α-glucosidase, α-amylase, acetyl- and butyrylcholinesterase-inhibiting, Gram-positive/-negative antimicrobial, cytotoxic, and genotoxic activities. A prominent apolar bioactive compound zone was tentatively assigned to fatty acids, in particular linolenic acid, via electrospray ionization high-resolution mass spectrometry. The detected antioxidant, antimicrobial, antidiabetic, anticholinesterase, cytotoxic, and genotoxic potentials of these vegetable plants, in particular C. rubens, S. biafrae, and S. macrocarpon, may validate some of their ethnomedicinal uses.

Phenolic Content Analysis of Two Species Belonging to the Lamiaceae Family: Antioxidant, Anticholinergic, and Antibacterial Activities.

The Lamiaceae family are utilized as ornamental, medicinal, and food supplements throughout the world. The current study focuses on a comparative analysis of the phenolic compositions and bioactivities (including antioxidant, anticholinergic, and antibacterial activities) of ethanolic extracts derived from the aerial parts of the two species (Lavandula stoechas L. and Thymus sipyleus Boiss). The presence of phenolic compounds and phytochemicals in the plant extracts was identified using the LC-MS/MS technique. The LC-MS/MS analysis revealed that vanillic acid (125,596.66 µg/L) was the most abundant phytochemical in L. stoechas. Kaempferol (8550.52 µg/L) was the most abundant substance in Thymus sipyleus. The assessment of the antioxidant efficacy of the species extracts was conducted using the DPPH (2.2-diphenyl-1-picryl-hydrazyl-hydrate), ABTS (2.2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)), Fe3+-Fe2+ reducing, and CUPRAC (Cu2+-Cu+ reducing) assays. The anticholinergic activity of the samples was determined using the acetylcholinesterase (AChE) inhibition assay. The results of antioxidant activity were higher in the T. sipyleus than in the L. stoechas ethanol extracts. The extracts of L. stoechas exhibited radical scavenging activity ranging from 15 to 18%, while T. sipyleus had activity effects ranging from 34% to 38%. The AChE inhibition potential for L. stoechas and T. sipyleus extracts as IC50 values were 0.221 ± 0.01 mg/mL and 0.067 ± 0.02 mg/mL, respectively. The antibacterial effects of the ethanolic extracts of these species against pathogenic bacteria isolates were determined using the MIC (minimal inhibitory concentration) method. These findings indicated that the extracts from L. stoechas and T. sipyleus possess the potential to be natural antioxidants in the realm of food preservation. Additionally, their antioxidant, anticholinergic, and antimicrobial properties suggest potential therapeutic utility in the management of certain diseases.

Inhibitory capacity of extracts and main constituents of hop flowers

BackgroundHops (Humulus lupulus Linn.) produce flowers that are used on an industrial scale to impart sensory properties to beer and have some pharmacologic properties already published in the literature. Gout, type 2 diabetes and Alzheimer’s are diseases the appearance/aggravation of which is related to the action of enzymes such as xanthine oxidase, α-glucosidase and acetylcholinesterase, respectively, which are associated with oxidative stress. Our hypothesis is that the extracts and chemical constituents isolated from the flowers of hops act as enzyme inhibitors and scavenge free radicals. Hexane and methanol extracts were obtained from flowers of different cultivars produced in Brazil and from commercial pellets, with the contents of bitter acids and xanthohumol determined by high-performance liquid chromatography (HPLC), after QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) extraction and were evaluated for their pharmacologic properties.ResultsThe extracts and analytical standards showed a significant capacity against oxidative stress in all evaluated methods (DPPH·, ABTS·+, HO·). Xanthohumol and extracts with a high concentration of this compound demonstrated a strong capacity to inhibit the xanthine oxidase and significantly reduce the formation of uric acid. The results with the α-glucosidase revealed promising pharmacologic action for all samples, even more effective than the commercial drug (acarbose). The acetylcholinesterase inhibition assays were effective for hexane extracts and ICE-4, with values obtained close to the eserine.ConclusionIt can be concluded with the results obtained that extracts and patterns of hop flowers showed promising potential to combat oxidative stress and complementary treatment of gout, type 2 diabetes and Alzheimer’s disease.

One-step microwave enhanced synthesis, biological evaluation, and molecular modeling investigations of donepezil analogs as acetylcholinesterase inhibitors

Acetylcholinesterase (AChE) is an important enzyme that hydrolyzes the neurotransmitter acetylcholine. Inhibiting AChE represents one of the therapeutic strategies for treating the symptoms of Alzheimer’s disease (AD). To date, only several AD drugs have been approved by the U.S. Food and Drug Administration, with donepezil being the most commonly used. In this study, we synthesized 46 donepezil-based analogs via a one-step microwave-assisted synthetic route and evaluated them in vitro using an AChE inhibition assay in order to better understand structural requirements important for the inhibition. Our structure–activity relationship studies revealed that various halogens, electron-withdrawing, and electron-donating groups placed in the ortho- and meta-positions on the phenyl moiety of donepezil are generally well-tolerated, yielding potent AChE inhibitors in the low nanomolar range similar to donepezil, while polysubstitutions led to moderate or significant decreases in inhibition potency.

Bioassays guided isolation of berberine from Berberis lycium and its neuroprotective role in aluminium chloride induced rat model of Alzheimer's disease combined with insilico molecular docking.

Berberis lycium is an indigenous plant of Pakistan that is known for its medicinal properties. In the current study, we investigated the anti-Alzheimer's effect of berberine isolated from Berberis lycium. Root extract of B. lycium was subjected to acetylcholinesterase inhibition assay and column chromatography for bioassays guided isolation of a compound. The neuroprotective and memory improving effects of isolated compound were evaluated by aluminium chloride induced Alzheimer's disease rat model, elevated plus maze (EPM) and Morris water maze (MWM) tests., Levels of dopamine and serotonin in rats brains were determined using HPLC. Moreover, western blot and docking were performed to determine interaction between berberine and β-secretase. During fractionation, ethyl acetate and methanol (3:7) fraction was collected from solvent mixture of ethyl acetate and methanol. This fraction showed the highest anti-acetylcholinesterase activity and was alkaloid positive. The results of TLC and HPLC analysis indicated the presence of the isolated compound as berberine. Additionally, the confirmation of isolated compound as berberine was carried out using FTIR and NMR analysis. In vivo EPM and MWM tests showed improved memory patterns after berberine treatment in Alzheimer's disease model. The levels of dopamine, serotonin and activity of antioxidant enzymes were significantly (p<0.05) enhanced in brain tissue homogenates of berberine treated group. This was supported by decreased expression of β-secretase in berberine treated rat brain homogenates and good binding affinity of berberine with β-secretase in docking studies. Binding energies for interaction of β-secretase with berberine and drug Rivastigmine is -7.0 kcal/mol and -5.8 kcal/mol respectively representing the strong interactions. The results of docked complex of secretase with berberine and Rivastigmine was carried out using Gromacs which showed significant stability of complex in terms of RMSD and radius of gyration. Overall, the study presents berberine as a potential drug against Alzheimer's disease by providing evidence of its effects in improving memory, neurotransmitter levels and reducing β-secretase expression in the Alzheimer's disease model.

Chromatographic and Biological Screening of Chosen Species of Schisandraceae Family: Schisandra chinensis, S. rubriflora, S. sphenanthera, S. henryi and Kadsura japonica.

HPLC and TLC profiling was carried out for leaf and fruit extracts of five Schisandraceae species: Schisandra chinensis, S. rubriflora, S. spehenanthera, S. henryi and Kadsura japonica. HPLC measurements confirmed presence of lignans and phenolic compounds in fruits and leaves of all tested species. The most abundant in lignans was S. chinensis fruit extract in which 15 compounds were detected (e. g.: schisandrol A, schisanhenol, γ-schisandrin, gomisin N). The effect-directed detection, i. e., TLC-direct bioautography against Bacillus subtilis, showed exceptionally high activity for S. chinensis and S. rubriflora fruit extracts. On the other hand, TLC-DB enzyme tests (α-glucosidase, lipase, tyrosinase and acetylcholinesterase (AChE) inhibition assays) showed that all fruit and leaf extracts have ability to inhibit the above-mentioned enzymes (except for the K. japonica fruit). The leaf extracts showed much stronger antioxidant activity than the fruit ones, which were assessed and compared using both TLC-direct bioautography and spectrophotometric measurements based on ABTS, DPPH and FRAP tests.

Bioactivity profiles of six baobab fruit pulp powders via planar chromatography hyphenated with effect-directed analysis

Baobab (Adansonia digitata) fruit pulp has a high nutrient content and has been traditionally used for medicinal purposes (e.g., as an anti-inflammatory and antioxidant agent) that may help protect against chronic diseases. Six different baobab fruit pulp powders were investigated using three different extractants and analyzed by high-performance thin-layer chromatography (HPTLC) hyphenated with antibacterial bioassays and enzyme inhibition assays. The developed non-target effect-directed screening was performed after extraction with pentyl acetate – ethanol 1:1 (V/V) on the HPTLC plate silica gel 60 using toluene – ethyl acetate – methanol 6:3:1 (V/V/V) as mobile phase system and derivatization via the anisaldehyde sulfuric acid reagent for detection. The physico-chemical profiles of the six baobab fruit pulp powder extracts were comparable, although the intensity of some zones was moderately different. The following effect-directed profiling via tyrosinase, α-glucosidase, and acetylcholinesterase inhibition assays as well as antibacterial Aliivibrio fischeri and Bacillus subtilis bioassays revealed one prominent multipotent bioactive compound zone in common, more or less active in all five studied (bio)assays. Via the recording of high-resolution mass spectra, this compound zone was tentatively assigned to coeluting saturated (palmitic acid 16:0 and stearic acid 18:0), monounsaturated (oleic acid 18:1), and polyunsaturated (linoleic acid 18:2 and linolenic acid 18:3) fatty acids. This finding was confirmed by other studies, which already proved individual activities of fatty acids. The first (bio)activity profiling of baobab fruit pulp powders via HPTLC-effect-directed analysis revealed that the baobab fruit could be considered as a functional food, however, further research is needed to study the impact on health and the influences on the bioactivity arising from different climates, years and soils or regions.

Therapeutic Potential of Green-Engineered ZnO Nanoparticles on Rotenone-Exposed D. melanogaster (Oregon R+): Unveiling Ameliorated Biochemical, Cellular, and Behavioral Parameters.

Nanotechnology holds significant ameliorative potential against neurodegenerative diseases, as it can protect the therapeutic substance and allow for its sustained release. In this study, the reducing and capping agents of Urtica dioica (UD), Matricaria chamomilla (MC), and Murraya koenigii (MK) extracts were used to synthesize bio-mediated zinc oxide nanoparticles (ZnO-NPs) against bacteria (Staphylococcus aureus and Escherichia coli) and against rotenone-induced toxicities in D. melanogaster for the first time. Their optical and structural properties were analyzed via FT-IR, DLS, XRD, EDS, SEM, UV-Vis, and zeta potential. The antioxidant and antimicrobial properties of the fabricated ZnO-NPs were evaluated employing cell-free models (DPPH and ABTS) and the well diffusion method, respectively. Rotenone (500 µM) was administered to Drosophila third instar larvae and freshly emerged flies for 24-120 h, either alone or in combination with plant extracts (UD, MC, an MK) and their biogenic ZnO-NPs. A comparative study on the protective effects of synthesized NPs was undertaken against rotenone-induced neurotoxic, cytotoxic, and behavioral alterations using an acetylcholinesterase inhibition assay, dye exclusion test, and locomotor parameters. The findings revealed that among the plant-derived ZnO-NPs, MK-ZnO NPs exhibit strong antimicrobial and antioxidant activities, followed by UD-ZnO NPs and MC-ZnO NPs. In this regard, ethno-nano medicinal therapeutic uses mimic similar effects in D. melanogaster by suppressing oxidative stress by restoring biochemical parameters (AchE and proteotoxicity activity) and lower cellular toxicity. These findings suggest that green-engineered ZnO-NPs have the potential to significantly enhance outcomes, with the promise of effective therapies for neurodegeneration, and could be used as a great alternative for clinical development.

Neurotoxicity in complex environmental mixtures-a case-study at River Danube in Novi Sad (Serbia) using zebrafish embryos.

Acetylcholinesterase (AChE) inhibitors are an important class of neuroactive chemicals that are often detected in aquatic and terrestrial environments. The correct functionality of the AChE enzyme is linked to many important physiological processes such as locomotion and respiration. Consequently, it is necessary to develop new analytical strategies to identify harmful AChE inhibitors in the environment. It has been shown that mixture effects and oxidative stress may jeopardize the application of in vivo assays for the identification of AChE inhibitors in the environment. To confirm that in vivo AChE assays can be successfully applied when dealing with complex mixtures, an extract from river water impacted by non-treated wastewater was bio-tested using the acute toxicity fish embryo test (FET) and AChE inhibition assay with zebrafish. The zebrafish FET showed high sensitivity for the extract (LC10 = relative extraction factor 2.8) and we observed a significant inhibition of the AChE (40%, p < 0.01) after 4-day exposure. Furthermore, the extract was chromatographically fractionated into a total of 26 fractions to dilute the mixture effect and separate compounds according to their physico-chemical properties. As expected, non-specific acute effects (i.e., mortality) disappeared or evenly spread among the fractions, while AChE inhibition was still detected in five fractions. Chemical analysis did not detect any known AChE inhibitors in these active fractions. These results confirm that the AChE assay with Danio rerio can be applied for the detection of neuroactive effects induced in complex environmental samples, but also, they highlight the need to increase analytical and identification techniques for the detection of neurotoxic substances.

Ameliorative effect of scopolamine-induced cognitive dysfunction by Fufangmuniziqi formula: The roles of alkaloids, saponins, and flavonoids

Ethnopharmacological relevanceFufangmuniziqi formula (FFMN), a traditional Uyghur medicine used in China, is derived from an ancient Uyghur medical book and consists of 13 herbs. The herbs of FFMN, such as Peganum harmala L., Glycyrrhiza uralensis Fisch., and Nigella glandulifera, have been demonstrated to have acetylcholinesterase (AChE) inhibitory, anti-neuroinflammatory, or antioxidant effects. Therefore, FFMN may have a good anti-Alzheimer's disease (AD) effect, but its specific action and mechanism need to be further proven. Aim of the studyThis study aims to investigate the anti-AD effects of FFMN and the role played by alkaloids, flavonoids, and saponins in anti-AD. Materials and methodsThe alkaloids, flavonoids, and saponins fractions of FFMN were prepared by macroporous resin chromatography. The absorbed ingredients in the drug-containing serum were identified by UPLC⁃Q⁃TOF⁃MS. An AD mouse model was established by intraperitoneal injection of scopolamine (SCO). The role of different fractions of FFMN in the anti-AD process was examined by Morris water maze (MWM), in-vitro cell, and AChE inhibition assay. ResultsA total of 20 ingredients were identified in the serum samples collected after oral administration of FFMN, and seven compounds were selected as candidate active compounds. MWM experiments showed that different fractions of FFMN could significantly improve SCO-induced learning memory impairment in mice. The alkaloids fraction (ALK) regulated cholinergic function by inhibiting AChE activity, activating choline acetyltransferase activity, and protein expression. Flavonoids and saponins were more potent than the ALK in downregulating pro-inflammatory factors or inflammatory mediators, such as TNF-α, MPO, and nitric oxide. Western blot results further confirmed that flavonoids and saponins attenuated neuroinflammation by inhibiting the phosphorylation of IκB and NF-κB p65. This result was also verified by in-vitro cellular assays. FFMN enhanced antioxidant defense by increasing the activity of superoxide dismutase and reducing the production of MDA. Combined with cellular experiments, flavonoids and saponins were proven more protective against oxidative damage. ConclusionFFMN improved cognitive and memory impairment in the SCO-induced AD mouse model. ALK mainly enhanced the function of the cholinergic system. Flavonoid and saponin fractions mainly attenuated neuroinflammation and oxidative stress by modulating the NF-κB pathway. All these findings strongly suggested that the combination of alkaloid, flavonoid, and saponin fractions derived from FFMN is a promising anti-AD agent that deserves further development.

Neuroprotective and nephroprotective effects of Ircinia sponge in polycyclic aromatic hydrocarbons (PAHs) induced toxicity in animal model: a pharmacological and computational approach

The present study investigated the neuroprotective and nephroprotective effects of the sponge Ircinia sp. ethyl acetate extract (ISPE) against persistent aromatic pollutants in vitro and in vivo. Different exponential experimental assays were applied to this study. An in vitro study to investigate the potential therapeutic effect of ISPE using antioxidants (for example, ABTS and DPPH) and anti-Alzheimer assays (inhibition of acetylcholinesterase); the in-vivo study was designed to evaluate the protective effect of ISPE as neuroprotective and nephroprotective against the destructive effect of PAH. Several assays included oxidative assays (LPO), antioxidant biomarkers (GSH, GST), and inflammatory and neurodegenerative biomarkers (PTK,SAA). Additionally, the results were confirmed using histopathological examination. The in silico screening study improved the in vitro and in vivo findings through interaction between the aryl hydrocarbon receptor (AHR) and the polyphenolic content of ISPE extract, which was determined using LCMSM. The results and discussion showed that ISPE exhibited a promising antioxidant and anti-acetylcholinesterase activity as evidenced by IC50 values of 49.74, 28.25, and 0.18 µg/mL in DPPH, ABTS, and acetylcholinesterase inhibition assays, respectively. In vivo, the study showed that animals receiving ISPE before poly aromatic hydrocarbons administration PAHs (Prot, ISPE) showed significant amelioration in kidney functions manifested by the reduction of serum urea, uric acid, and creatinine by 40.6%, 66.4%, and 134.8%, respectively, concerning PAH-injected mice (HAA). Prot, ISPE revealed a decline in malondialdehyde (MDA) and total proteins (TP) in kidney and brain tissues by 73.63% and 50.21%, respectively, for MDA and 59.82% and 80.41%, respectively, for TP with respect to HAA. Prot, ISPE showed significant elevation in reduced glutathione (GSH) and glutathione transferase (GST) in kidney and brain tissues and reduction in the inflammatory and pre-cancerous biomarkers, namely, serum protein tyrosine kinases (PTKs) and serum amyloid A (SAA). These findings were further supported by histopathological examination of kidney and brain tissues, which revealed normal structure approaching normal control. Metabolic profiling of ISPE using LC–MS-MS showed the presence of fourteen polyphenolic compounds belonging mainly to phenolic acids and flavonoids. In silico study revealed that all the tested compounds exerted certain binding with the aryl hydrocarbon receptor, where rutin showed the best fitting (ΔG = − 7.6 kcal/mol−1) with considerable pharmacokinetic and pharmacodynamic properties revealed from in silico ADME (Absorption, Distribution, Metabolism, and Excretion) study. Hence, it can be concluded that the Ircinia sponge showed a promising protective effect versus kidney and brain toxicity triggered by PAHs.Graphical

Quantitative-HPLC-DAD polyphenols analysis, anxiolytic and cognition enhancing potentials of Sorbaria tomentosa Lindl. Rehder

Ethnopharmacological relevanceMedicinal plants of the family Rosaceae have a long history of traditional uses in the management of neurological disorders. Sorbaria tomentosa Lindl. Rehder is composed of antioxidant and neuroprotective polyphenolics. Aims of the studyThe current study was designed to explore phenolics profile via high performance liquid chromatography-photodiode array detector (HPLC-DAD) and validated the neuroprotective and anxiolytic potentials of S. tomentosa by applying in vitro and in vivo approaches. Materials and methodsThe plant crude methanolic extract (St.Crm) and fractions were subjected to HPLC-DAD analysis for qualitative and quantitative assessment of phytochemicals. Samples were screened for in vitro free radicals scavenging assays by using 2,2-diphenylpicrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) along with acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes inhibition assays. For cognitive and anxiolytic studies, mice were subjected to open field, elevated plus maze (EPM), light-dark model, Y-maze, shallow water maze (SWM), and novel object recognition (NOR) tests. ResultsHPLC-DAD analysis revealed the presence of high concentrations of phenolic compounds. For instance, in St.Cr, 21 phenolics were quantified, among which apigenin-7-glucoside (291.6 mg/g), quercetin (122.1 mg/g), quercetin-3-feruloylsophoroside-7-glucoside (52.6 mg/g), quercetin-7-glucoside (51.8 mg/g), ellagic acid (42.7 mg/g), luteolin (45.0 mg/g), kaempferol (40.5 mg/g), 5-feruloylquinic acid (43.7 mg/g) were present in higher concentrations. Likewise, in ethyl acetate fraction (St.Et.Ac), 21 phenolics were identified as 3,5-di-caffeoylquinic acid (177.4 mg/g) and 5-hydroxybenzoylquinic acid (46.9 mg/g) were most abundant phytochemicals. Highly valuable phenolics were also identified in other fractions including butanol (St.Bt), chloroform (St.Chf), and n-hexane (St.Hex). The various fractions exhibited concentration dependent inhibition of free radicals in DPPH and ABTS assays. Potent AChE inhibitory potentials were revealed by the test samples with St.Chf, St.Bt and St.EtAc being the most active having an IC50 of 298.1, 580.1, and 606.47 μg mL−1, respectively. Similarly, St.Chf, St.Bt, St.EtAc and St.Cr exhibited potent BChE inhibitory activity and was observed as 59.14, 54.73, 51.35 and 49.44%, respectively. A significant improvement in the exploratory behavior was observed in open field test and stress/anxiety was relieved effectively at 50-100 mg/kg. Likewise, EPM, light-dark and NOR tests revealed an anxiolytic and memory enhancing behaviors. These effects were further corroborated from the Y-maze and SWM transgenic studies that showed considerable improvement in cognition retention. ConclusionsThese findings concluded that S. tomentosa possessed potential anxiolytic and nootropic efficacies and may have therapeutic potential in neurodegenerative disorders.

Comparative Impact of Solvent Extracts of Spondias mombin Leaves on In-vitro Antioxidant and Acetylcholinesterase Inhibitory Activities

The underlying cause of a number of neurological disorders is oxidative stress. Given the dearth of medications now available to treat such disorders and their accompanying detrimental impacts, an urgent need exists for the global identification of brand-new antioxidants and acetylcholinesterase (AChE) inhibitors. This study evaluated the comparative impacts of the antioxidant and acetylcholinesterase activities of n-hexane, ethyl acetate, and methanol extracts of S. mombin leaves. The dried leaf samples of the plant were triturated. Following maceration of the powdered plant materials in each of the three extraction solvents (methanol, n-hexane, and ethyl acetate), the resulting solutions were separately subjected to lyophilization. The in-vitro antioxidant analysis was determined by employing the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and lipid peroxidation (LPO) assays. Acetylcholinesterase (AChE) inhibitory abilities of the various extracts were also evaluated using a standard protocol. The methanol extract showed the greatest DPPH scavenging (75.91%) and AChE inhibitory (40.17%) properties in the DPPH and AChE inhibitory assays, respectively. The DPPH and lipid peroxidation assays also demonstrated that all extracts had dose-dependent antioxidant properties. In addition, evaluations of each extract's ability to inhibit lipid peroxidation revealed that, at initial concentrations of 0.05 mg/L and 0.1 mg/L, the methanol extract displayed stronger LPO inhibitory effects (5.90%, 6.15%) than the n-hexane (4.00%, 5.4%) and ethyl acetate (3.26%, 3.99%) extracts. Nonetheless, the n-hexane extract showed a greater LPO inhibitory effect (10.00%, 10.34%) at higher dosages of 0.2 mg/L and 0.5 mg/L respectively. The results of this study have shown that methanol is the best solvent for exploiting the pharmacological benefits of S. mombin leaves thereby reasserting the numerous applications of the plant in traditional medicine. It has also paved the way for the development of novel therapeutic alternatives for a range of neurodegenerative disorders and other health concerns.

A Study of the Chemical Composition and Biological Activity of Michelia macclurei Dandy Heartwood: New Sources of Natural Antioxidants, Enzyme Inhibitors and Bacterial Inhibitors.

The wood of Michelia macclurei Dandy (MD) is an excellent material that is widely used in the furniture, handicraft, and construction industries. However, less research has been conducted on the chemical composition and biological activity of heartwood, which is the main valuable part of the wood. This study aimed to investigate the chemical composition and biological activities of the heartwood of Michelia macclurei Dandy (MDHW) and to confirm the active ingredients. Triple quadrupole gas chromatography-mass spectrometry (GC-MS) was used to characterize the volatile components of MDHW, while ultra-performance liquid chromatography-mass spectrometry was used to analyze the non-volatile components (UPLC-MS). The total reducing power, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging assays, acetylcholinesterase and α-glucosidase inhibition assays, and an antimicrobial test of 4 gram bacteria were used to describe the in vitro bioactivities. The GC-MS analysis showed that the volatile components of MDHW were mainly fatty compounds and terpenoids, with sesquiterpenes and their derivatives dominating the terpene composition. β-elemene was the main terpene component in the steam distillation (11.88%) and ultrasonic extraction (8.2%) methods. A total of 67 compounds, comprising 45 alkaloids, 9 flavonoids, 6 lignans, and others, were found by UPLC-MS analysis. The primary structural kinds of the non-volatile components were 35 isoquinoline alkaloids. Alkaloids were the predominant active constituent in all MDHW extracts, including crude extracts, alkaloid fractions, and non-alkaloid fractions. These extracts all demonstrate some biological effects in terms of antioxidant, enzyme inhibition, and bacterial inhibition. The findings of this study show that MDHW is abundant in chemical structure types, has great bioactivity assessment, and has the potential to be used to create natural antioxidants, products that postpone Alzheimer's disease and lower blood sugar levels and antibacterial agents.

Near-infrared-excitable acetylcholinesterase-activated fluorescent probe for sensitive and anti-interference detection of pesticides in colored food

The development of a common and anti-interference acetylcholinesterase (AChE) inhibition assay for plant-originated food samples has been of great challenge because of the prevalent and strong signal interferences from natural pigments. Plant pigments normally exhibit non-negligible absorbance in the UV-visible region. As a result, the signals of a typical near-infrared (NIR) fluorescent probe could be disturbed through primary inner filter effect if it is excited by UV-visible light during plant sample analysis. In this work, an NIR-excitable AChE-activated fluorescent probe was biomimetically designed and synthesized. And the NIR-excitation strategy was utilized for the anti-interference detection of organophosphate and carbamate pesticides in colored samples with this probe. Sensitive and rapid response to AChE and pesticides was achieved due to the high affinity of the biomimetic recognition unit in the probe. The limits of detection for four representative pesticides including dichlorvos, carbofuran, chlorpyrifos and methamidophos reached 0.0186 μg/L, 2.20 μg/L, 12.3 μg/L and 13.6 μg/L, respectively. Most importantly, fluorescent response to pesticide contents could be accurately measured in the coexistence of different plant pigments by this probe, and the measured results showed completely irrelevance to the plant pigments and their colors. Taking advantage of such probe, the new developed AChE inhibition assay showed good sensitivity and anti-interference ability in the detection of organophosphate and carbamate pesticides in real samples.

In vitro and in silico analysis of the Anopheles anticholinesterase activity of terpenoids.

Anopheles gambiae, An. coluzzii, An. arabiensis, and An. funestus are major vectors in high malaria endemic African regions. Various terpenoid classes form the main chemical constituent repository of essential oils, many of which have been shown to possess insecticidal effects against Anopheles species. The current study aimed to assess the bioactivity of terpenoids including four sesquiterpene alcohols, farnesol, (-)-α-bisabolol, cis-nerolidol, and trans-nerolidol; a phenylpropanoid, methyleugenol, and a monoterpene, (R)-(+)-limonene, using the larvicidal screening assay against the four Anopheles species. The mechanism of action was investigated through in vitro acetylcholinesterase inhibition assay and in silico molecular modelling. All six terpenoids showed potent larvicidal activity against the four Anopheles species. Insights into the mechanism of action revealed that the six terpenoids are strong AChE inhibitors against An. funestus and An. arabiensis, while there was a moderate inhibitory activity against An. gambiae AChE, but very weak activity against An. coluzzii. Interestingly, in the in silico study, farnesol established a favourable hydrogen bonding interaction with a conserved amino acid residue, Cys447, at the entrance to the active site gorge. While (-)-α-bisabolol and methyleugenol displayed a strong interaction with the catalytic Ser360 and adjacent amino acid residues; but sparing the mutable Gly280 residue that confers resistance to the current anticholinesterase insecticides. As a result, this study identified farnesol, (-)-α-bisabolol, and methyleugenol as selective bioinsecticidal agents with potent Anopheles AChE inhibition. These terpenoids present as natural compounds for further development as anticholinesterase bioinsecticides.