Inhibition Of Cholinesterase Activity Research Articles

New tetrahydro-isoquinoline derivatives as cholinesterase and α-glycosidase inhibitors: Synthesis, characterization, molecular docking & dynamics, ADME prediction, in vitro cytotoxicity and enzyme inhibition studies

In the presented work, it was reported about the synthesis procedure of new isoquinoline derivatives starting from chalcones and acetoacetanilide, a multi-step synthesis pathway yielded four isoquinoline derivatives (5a-d). At the first stage of our research, corresponding cyclohexanone derivatives are synthesized from the Michael addition reaction of chalcones with acetoacetanilide. After that, on the based cyclohexanone derivative was obtained novel isoquinoline derivatives. According to the structures, a plausible reaction mechanism was suggested. The structure of the synthesized compounds was confirmed by NMR spectroscopy method and X-Ray analysis. Additionally, to determine cholinesterase and α-glycosidase inhibition activity compounds 5(a-d) were tested against AChE, BChE and α-Gly as in vitro. In this work, the novel isoquinoline derivatives 5(a-d) demonstrated encouraging inhibitory action against α-glucosidase, as seen by their IC50 values ranging from 9.95 to 19.25 nM. This indicates that they were approximately 1–3 times more active than acarbose (IC50 = 23.07 nM). Also, the molecular docking IFD XP Gscores, dynamics simulations and MM-GBSA ΔG binding free energies of the isoquinoline derivatives on hAChE, hBChE and α-Gly are determined. The molecular dynamics (MD) simulations conducted in this study provided valuable insights into the dynamic behaviors, stabilities, and interaction profiles of ligand–protein complexes involving the R and S isomers of compounds 5c-hAChE, 5c-hBChE, and 5a-αGly. The MD simulations were crucial for understanding the complex stabilities and dynamic mechanisms of the ligand–protein interactions, shedding light on the potential inhibitory roles of these compounds..

In vitro Cholinesterase Inhibitory Activities of Piper betle Stem and their Correlation with In silico Docking Studies of its Phytoconstituents

Background: Worldwide, millions of people are affected by neurodegenerative diseases. Even though treatment may help to reduce some of the mental or physical symptoms connected with neurodegenerative diseases, there is at present no way to slow disease development and no recognized cure. Objective: The current study was carried out to explore the cholinesterase (ChE) inhibitory properties of the stem of Piper betle and correlate them with in silico docking results of its phytoconstituents. Methods: The dried Piper betle stem was used to be extracted with purified water by using the maceration technique. The resultant was assessed for ChE inhibitory activity using Ellman’s method. The inhibitory profile of the aqueous extract of Piper betle (AEPB) stem was compared with rivastigmine, which is a standard cholinesterase inhibitor. The phytoconstituents of AEPB were procured from various literature studies. In silico docking studies were carried out with the help of AutoDock 4.2 software. Results: AEPB considerably inhibited AChE and BuChE with the inhibition constant values of 0.437 ± 0.62 μg/ml and 0.371 ± 0.62 μg/ml, respectively, in a dose-dependent manner. In computational evaluation, the selected phytoconstituents exhibited excellent binding interactions prevailing with cholinesterase targets than the standard rivastigmine. Conclusion: Based on the in vitro and in silico evaluations, Piperol A showed notable concentrationdependent inhibition of AChE and BuChE. These in vitro analyses and molecular docking studies will be helpful for the development of potent ChE inhibitors for the management of Alzheimer’s disease.

Development of novel 9H-carbazole-4H-chromene hybrids as dual cholinesterase inhibitors for the treatment of Alzheimer's disease.

Alzheimer's disease (AD) is a neurodegenerative disease affecting mental ability and neurocognitive functions. Cholinesterase enzymes affect concentration of acetylcholine in the brain, leading to dementia. Thus, there is an urgent need to develop novel dual cholinesterase inhibitors as possible anti-AD drugs. Herein, we have designed and synthesized a novel series of 9H-carbazole-4H-chromenes 4(a-l) through a one-pot three-component reaction of salicylaldehydes (1), hydroxycarbazole (2) and N-methyl-1-(methylthio)-2-nitroethenamine (3) using triethylamine as a catalyst in ethanol. Synthetic transformation involves the formation of two C-C bonds and one C-O bond in a single step to obtain desired analogs. The rapid one-pot reaction does not require chromatographic purification, proceeds under mild conditions, and exhibits good tolerance toward various functional groups with high synthetic yields. Synthesized compounds were screened for cytotoxicity using MTT assay in BV-2 microglial cells. These compounds were then in-vitro screened against acetylcholinesterase (AChE) and butyrylcholinestrase (BuChE) enzymes. Most of these ligands have shown dual cholinesterase inhibitory activity compared to the standard drug. In-vitro results showed that the compounds 4a and 4d have promising anticholinesterase response against both cholinesterase enzymes (4a, AChE IC50: 5.76µM, BuChE IC50: 48.98µM; 4d, AChE IC50: 3.58µM, BuChE IC50: 42.73µM). In-vitro results were validated by molecular docking and dynamic simulation at 100ns. Molecular docking and molecular dynamics simulation study strongly supported structural features present in these analogs. Together, these analogs could be exploited to develop dual anti-cholinesterase candidates to treat AD in combination with other drugs.

Lathyrane and premyrsinane Euphorbia diterpenes against Alzheimer's disease: Bioinspired synthesis, anti-cholinesterase and neuroprotection bioactivity

The first systematic acylated diversification of naturally scarce premyrsinane diterpenes, together with their biosynthetic precursors lathyrane diterpene were carried out. Two new series of premyrsinane derivates (1a-32a) and lathyrane derivates (1–32) were synthesized from the naturally abundant lathyrane diterpene Euphorbia factor L3 through a bioinspired approach. The cholinesterase inhibitory and neuroprotective activities of these diterpenes were investigated to explore potential anti-Alzheimer's disease (AD) bioactive lead compounds. In general, the lathyrane diterpenes showed the better acetylcholinesterase (AChE) inhibitory activity than that of premyrsinanes. The lathyrane derivative 17 bearing a 3-dimethylaminobenzoyl moiety showed the best AChE inhibition effect with the IC50 value of 7.1 μM. Molecular docking demonstrated that 17 could bond with AChE well (−8 kal/mol). On the other hand, premyrsinanes showed a better neuroprotection profile against H2O2-induced injury in SH-SY5Y cells. Among them, the premyrsinane diterpene 16a had significant neuroprotective effect with the cell viability rate of 113.5 % at 12.5 μM (the model group with 51.2 %). The immunofluorescence, western blot and reactive oxygen species (ROS) analysis were conducted to demonstrate the mechanism of 16a. Furthermore, a preliminary SAR analysis of the two categories of diterpenes was performed to provide the insights for anti-AD drug development.

Studies on the Effects of Fermentation on the Phenolic Profile and Biological Activity of Three Cultivars of Kale.

Fermentation is used not only to preserve food but also to enhance its beneficial effects on human health and achieve functional foods. This study aimed to investigate how different treatments (spontaneous fermentation or fermentation with the use of starter culture) affect phenolic content, antioxidant potential, and cholinesterase inhibitory activity in different kale cultivars: 'Halbhoner Grüner Krauser', 'Scarlet', and 'Nero di Toscana'. Chosen samples were further tested for their protective potential against the Caco-2 cell line. HPLC-MS analysis revealed that the fermentation affected the composition of polyphenolic compounds, leading to an increase in the content of rutin, kaempferol, sinapinic, and protocatechuic acids. In general, kale cultivars demonstrated various antioxidant activities, and fermentation led to an increase in total phenolic content and antioxidant activity. Fermentation boosted anti-cholinesterase activity most profoundly in 'Nero di Toscana'. Extracts of spontaneously fermented 'Scarlet' (SS) and 'Nero di Toscana' (NTS) showed cytoprotective properties, as revealed by the malondialdehyde (MDA), lactate dehydrogenase (LDH), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) assays. Additionally, strong anti-inflammatory activity of NTS was shown by decreased release of cytokines IL-1β and TNF-α. Collectively, the conducted studies suggest fermented kale cultivars as a potential source for functional foods.

Synthesis, biological activity, X-ray crystallographic, DFT calculations and molecular dynamics simulation studies of 2-phenylthiazole-1,3,5-triazine derivatives as potential cholinesterase inhibitors

Seven 2-phenylthiazole-1,3,5-triazine derivatives were synthesized and estimated for cholinesterase inhibitory activity. All target compounds were characterized by 1H NMR, 13C NMR, FT-IR, and HRMS, and their purity was tested by HPLC to be above 99%. The in vitro enzyme inhibitory activity assays determined that the majority of compounds exhibited moderate to good inhibitory effects on acetylcholinesterase or butyrylcholinesterase and compound 6g displayed the best acetylcholinesterase (IC50 = 0.26 μM) and butyrylcholinesterase inhibitory activity (IC50 = 0.78 μM). The structures of compounds 6a and 6g was further investigated by single-crystal X-ray diffraction, and the Hirshfeld surfaces, crystal voids, and molecular energy framework analysis were further discussed to study the molecular conformations, crystal packing modes, intermolecular interactions, and their energies. What's more, the density functional theory calculations including frontier molecular orbitals, global reactivity descriptors, and molecular electrostatic potential were carried out to reveal the chemical reactivity and stability of 6a and 6g molecules. Molecular docking study indicated that compound 6g could not only interact with acetylcholinesterase but also with butyrylcholinesterase. Molecular dynamics simulations study showed that compound 6g could bind to acetylcholinesterase stably. In addition, ADMET properties prediction implied the drug-like properties and safety of seven compounds. Considering these results, several of the target compounds could work as novel cholinesterase inhibitors, and compound 6g may possess the potential to become a lead compound to develop novel anti-Alzheimer drugs.

Lycopodium alkaloids from Huperzia serrata and their cholinesterase inhibitory activities

Huperzia serrata, belonging to the Lycopodiaceae family, has been traditionally utilized for the management of treating rheumatic numbness, arthritic pain, dysmenorrhea, and contusions. This plant is a rich source of lycopodium alkaloids, some of which have demonstrated notable cholinesterase inhibitory activity. The objective of this study was to identify lycopodium alkaloids with cholinesterase inhibitory properties from H. serrata. The structures of these alkaloids were elucidated by HRESIMS, NMR (including a 1H−15N HMBC experiment), ECD methods and single-crystal X-ray diffraction. The inhibitory activities against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) were assessed using a modified Ellman's method. Consequently, sixteen lycopodium alkaloids (1–16), including ten previously undescribed ones named huperradines A–G and huperradines I–K (1–7 and 9–11), along with one previously undescribed naturally occurring compound, huperradine H (8), were isolated from H. serrata. Among these, compounds 7 and 1 exhibited potent and moderate AChE inhibition, with IC50 values of 0.876 ± 0.039 μM and 13.125 ± 0.521 μM, respectively. Our results suggest that huperradine G (7) may be a promising lead compound for the development of new AChE inhibitors for Alzheimer's disease (AD).

Insight into antioxidant-like activity and computational exploration of identified bioactive compounds in Talinum triangulare (Jacq.) aqueous extract as potential cholinesterase inhibitors

BackgroundRecent reports have highlighted the significance of plant bioactive components in drug development targeting neurodegenerative disorders such as Alzheimer’s disease (AD). Thus, the current study assessed antioxidant activity and enzyme inhibitory activity of the aqueous extract of Talinum triangulare leave (AETt) as well as molecular docking/simulation of the identified phytonutrients against human cholinesterase activities.MethodsIn vitro assays were carried out to assess the 2,2- azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) cation radicals and cholinesterase inhibitory activities of AETt using standard protocols. High performance liquid chromatography coupled with diode-array detection (HPLC–DAD) was employed to identify compounds in AETt. Also, for computational analysis, identified bioactive compounds from AETt were docked using Schrodinger's GLIDE against human cholinesterase obtained from the protein data bank (https://www.rcsb.org/).ResultsThe results revealed that AETt exhibited a significant concentration-dependent inhibition against ABTS cation radicals (IC50 = 308.26 ± 4.36 µg/ml) with butylated hydroxytoluene (BHT) as the reference. Similarly, AETt demonstrated a significant inhibition against acetylcholinesterase (AChE, IC50 = 326.49 ± 2.01 µg/ml) and butyrylcholinesterase (BChE, IC50 = 219.86 ± 4.13 µg/ml) activities with galanthamine as the control. Molecular docking and simulation analyses revealed rutin and quercetin as potential hits from AETt, having showed strong binding energies for both the AChE and BChE. In addition, these findings were substantiated by analyses, including radius of gyration, root mean square fluctuation, root mean square deviation, as well as mode similarity and principal component analyses.ConclusionOverall, this study offers valuable insights into the interactions and dynamics of protein–ligand complexes, offering a basis for further drug development targeting these proteins in AD.

Phytochemical Profiling and Biological Activities of Pericarps and Seeds Reveal the Controversy on "Enucleation" or "Nucleus-Retaining" of Cornus officinalis Fruits.

The fruits of Cornus officinalis are used not only as a popular health food to tonify the liver and kidney, but also as staple materials to treat dementia and other age-related diseases. The pharmacological function of C. officinalis fruits with or without seeds is controversial for treating some symptoms in a few herbal prescriptions. However, the related metabolite and pharmacological information between its pericarps and seeds are largely deficient. Here, comparative metabolomics analysis between C. officinalis pericarps and seeds were conducted using an ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry, and therapeutic effects were also evaluated using several in vitro bioactivity arrays (antioxidant activity, α-glucosidase and cholinesterase inhibitory activities, and cell inhibitory properties). A total of 499 secondary metabolites were identified. Thereinto, 77 metabolites were determined as key differential metabolites between C. officinalis pericarps and seeds, and the flavonoid biosynthesis pathway was identified as the most significantly different pathway. Further, 47 metabolites were determined as potential bioactive constituents. In summary, C. officinalis seeds, which demonstrated higher contents in total phenolics, stronger in vitro antioxidant activities, better α-glucosidase and butyrylcholinesterase inhibitory activities, and stronger anticancer activities, exhibited considerable potential for food and health fields. This work provided insight into the metabolites and bioactivities of C. officinalis pericarps and seeds, contributing to their precise development and utilization.

Current Progress on Central Cholinergic Receptors as Therapeutic Targets for Alzheimer's Disease.

Acetylcholine (ACh) is ubiquitously present in the nervous system and has been involved in the regulation of various brain functions. By modulating synaptic transmission and promoting synaptic plasticity, particularly in the hippocampus and cortex, ACh plays a pivotal role in the regulation of learning and memory. These procognitive actions of ACh are mediated by the neuronal muscarinic and nicotinic cholinergic receptors. The impairment of cholinergic transmission leads to cognitive decline associated with aging and dementia. Therefore, the cholinergic system has been of prime focus when concerned with Alzheimer's disease (AD), the most common cause of dementia. In AD, the extensive destruction of cholinergic neurons occurs by amyloid-β plaques and tau protein-rich neurofibrillary tangles. Amyloid-β also blocks cholinergic receptors and obstructs neuronal signaling. This makes the central cholinergic system an important target for the development of drugs for AD. In fact, centrally acting cholinesterase inhibitors like donepezil and rivastigmine are approved for the treatment of AD, although the outcome is not satisfactory. Therefore, identification of specific subtypes of cholinergic receptors involved in the pathogenesis of AD is essential to develop future drugs. Also, the identification of endogenous rescue mechanisms to the cholinergic system can pave the way for new drug development. In this article, we discussed the neuroanatomy of the central cholinergic system. Further, various subtypes of muscarinic and nicotinic receptors involved in the cognition and pathophysiology of AD are described in detail. The article also reviewed primary neurotransmitters that regulate cognitive processes by modulating basal forebrain cholinergic projection neurons.

Gastrodinol derivatives and prenylated flavones from the flower branch of Gastrodiaelata

Based on the research progress and traditional usage with whole herbal of the TCM "Tianma", chemical studies herein on the flower branch of Gastrodiaelata were carried out in-depth and got 13 compounds including the gastrodinols (1–4), the flavonoid morins (5–8, 11–12), together with the specialist mulberrofurans (9, 13) and gastrodiamide (10) for the first time from the species. The antibacterial and cholinesterase inhibitory activities were then evaluated and the results showed that compounds 5, 11, 12, 13 have good activity against anti-methicillin-resistant Staphylococcusaureus, and compounds 9, 13 had good acetylcholinesterase inhibitory activity. All these results provide new chemical composition for better understanding the traditional application of "Tianma" and for exploring new pharmacological ingredients.Graphical

Evaluation of Changes in Acetylcholinesterase Activity in Workers of Mehriz Elixir Pesticide Plant in Yazd

Background and Aims: Some pesticide chemical compounds, such as organophosphates and carbamates, interfere with or inhibit cholinesterase activity. Employees working in pesticide factories are one of the groups at risk of pesticide poisoning. This study aimed to evaluate the effect of work on exposure to toxins on the serum level of erythrocyte cholinesterase activity.
 Materials and Methods: This research was conducted on Mehriz Elixir Pesticide Factory workers. Blood samples were taken from 76 employees in 2 groups as a control group, and 38 workers were exposed to organophosphorus toxins three months after starting work in a factory. Cholinesterase activity was analyzed using the Elman method, and data were analyzed using SPSS software version 22.
 Results: The mean age of the subjects was 35.07 years, which was 35.26 in the exposure group (n = 38) and 34.89 in the control group (n = 38). The activity of acetylcholinesterase enzyme in the control group at 0, 10, and 20 minutes was 12.78 ku/l, 14.24 ku/l, and 15.45 ku/l, respectively. The acetylcholinesterase enzyme activity in the exposed group was 10.77 ku/l, 10.40 ku/l, and 10.36 ku/l at 0, 10, and 20 minutes, respectively. At all stages, the mean acetylcholinesterase activity in the control group was higher than the exposed group, but significant differences were observed at 10 and 20 minutes between the 2 groups.
 Conclusion: Exposure of workers to organophosphate inhibits acetylcholinesterase, which manifests by a decrease in the activity of this enzyme.

Studies on Secondary Metabolites and In vitro and In silico Anticholinesterases Activities of the Sea Urchin Echinometra mathaei Crude Venoms from the Persian Gulf-Bushehr

Background: Echinoderms are a unique source of amazing secondary metabolites with a wide spectrum of biological activities. Several species of sea urchins contain various toxins and biologically active metabolites. One of the most attractive approaches to treat Alzheimer's disease is searching for effective marine natural products with cholinesterase inhibitory activities. Objective: The current study is designed to investigate the in vitro and in silico acetylcholinesterase and butyrylcholinesterase inhibitory activities of the Persian Gulf echinoderm sea urchin Echinometra mathaei venom and related chemical compounds. Methods: The experiments for LD50, total protein, protein bands, in vitro cholinesterase inhibitory activities, the identity of secondary metabolites, and the in silico evaluations, respectively, were performed by Spearman-Karber, Lowry, SDS-PAGE, Ellman's spectroscopic, GC-MS, and docking methods. Results: The LD50 (IV rat) of the spine, gonad, and coelomic fluid from sea urchin samples were 2.231 ± 0.09, 1.03 ± 0.05, and 1.12 ± 0.13 mg/ml, respectively. The SDS-PAGE and total protein studies showed that at least a portion of the venom is protein in nature. GC-MS analysis of the identified samples revealed 12, 23, and 21 compounds with different chemical types, including alkaloids, terpenes, and steroids, respectively. According to the results, all samples act as significant inhibitors of both enzymes. In silico data for the identified compounds also confirmed the experimental results. Conclusion: The alkaloid compound 6H-Indolo[3,2,1-de] [1,5] naphthyridine-6-one,1,2,3a,4,5- hexahydro-8-hydroxy-3-methyl (C7) had the highest affinity for both enzymes. Further research is needed to determine whether C7 could be a therapeutic candidate for Alzheimer's disease.

Fatty Acid Composition, Antioxidant, and Enzyme Inhibition Activities of Cachrys Crassiloba (Boiss.) Meikle Fruit and Leaf Extracts

The genus Cachrys L. (Apiaceae) is distributed in the Mediterranean region and contains 11 species. C. crassiloba (Boiss.) Meikle is a species that grows in Southwest and Western Anatolia. Considering the literature studies, there were not enough studies on C. crassiloba. In this study, fatty acid composition, total phenol/flavonoid contents, antioxidant and acetyl-butrylcholinesterase, tyrosinase, and α-glucosidase enzyme inhibition activities of n-hexane and ethanol extracts prepared from C. crassiloba fruits and leaves were investigated by spectrophotometric methods at different concentrations. When the results were evaluated, it was found that the hexane extract of C. crassiloba leaf (123.92 ± 4.62 GA mg/g extract) had the highest total phenol content, while the ethanol extract of C. crassiloba fruit (134.38 ± 0.98 QE mg/g extract) had the highest flavonoid content. C. crassiloba hexane and ethanol leaf extracts (IC50= 8.04 ± 1.31 µg/mL; 10.30 ± 3.15 µg/mL) showed good antioxidant activity com-pared to the ascorbic acid (IC50= 14.59 ± 1.96 µg/mL) in DPPH assay. C. crassiloba leaf ethanol extract (IC50= 17.38 ± 5.02 µg/mL) has the highest ABTS scavenging activity. C. crassiloba extracts have mod-erate cholinesterase inhibitory activity. C. crassiloba leaf ethanol extract (IC50= 196.65 ± 1.94 µg/mL) has good tyrosinase enzyme inhibition activity. C. crassiloba leaf hexane extract (36.35% ± 1.13) was found to have significant inhibitory activity against α-glucosidase. In conclusion, besides its antioxidant activity, C. crassiloba may be effective against neurodegenerative diseases and skin disorders such as hyperpigmentation and diabetes mellitus, but further phytochemical analysis studies are required.

Phytoconstituents, antioxidant, and cholinesterase inhibitory activities of the leaves and stem extracts of Artocarpus sericicarpus

The study aimed to investigate the antioxidant and cholinesterase inhibitory activities of the leaves and stems of Artocarpus sericicarpus and to analyse the phenolic compounds in the extracts. The modified Ellman’s method was used to determine the cholinesterase inhibitory activities against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes. The antioxidant properties were evaluated using DPPH and ABTS methods. The total phenolic content (TPC) was measured by spectrometric assay, and compound identification was carried out by LC-MS/MS analysis. The results showed that the leaf and stem extracts of A. sericicarpus exerted significant inhibitory effects against AChE and BChE, as well as antioxidant activities. The stem ethanolic extract exhibited the highest potency against AChE and BChE with IC50 values of 5.81 and 11.46 µg/mL, respectively. The leaf and stem ethanolic extracts gave higher antioxidant activities and TPC compared to the water-based extracts. The LC-MS/MS analysis indicated the presence of phenolic compounds, such as flavones, flavonols, flavanones, prenylated chalcones, and xanthones in the extracts.

Synthesis, complexation, in vitro cholinesterase inhibitory activities and molecular docking of azinethiacrown ethers and acyclic thiacrown ethers derived indole

A series of new thiacrown ethers 14–16, 18–22 and acyclic thiacrown ethers (podands) 31–36 and 52–57 derived indole were synthesized and characterized by NMR and mass spectrometric techniques. Preliminary complexation study of podands 31–36 and 52–54 with some metal cations was studied using a conductometric method in acetonitrile at 25 °C. After the addition of acetonitrile solution of 31–36 or 52–54 podands to the metal cation acetonitrile solutions, the conductance was strongly decreased only in the presence of Hg2+ ions. The order of the stability constants of 1:1 podands–Hg2+ complexes is in the order: 52 > 53 > 35> 31> 32> 36> 33. On the other hand, podands 34 and 54 formed 1:2 podands–Hg2+ complexes.Thiacrown ethers 14–16, 18–22 and acyclic thiacrown ethers 31–36, and 52–57 were evaluated against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymes in vitro. The results indicated that compounds 14–57 demonstrated more potent inhibitory effects on AChE compared to BuChE, with IC50 values ranging from 0.244 to 1.359 mM and 1.270 to 81.473 mM, respectively. Notably, compound 16 most potently inhibited AChE with an IC50 value of 0.244 mM, being 149.6-fold more selective for AChE than BuChE. The presence of imine, ester groups, and sulfur, as well as oxygen or bromine atoms in the cyclic thiacrown ethers, improved their inhibitory activity against both AChE and BuChE.Inhibition of AChE by compounds 19, 20, 54, and 56 ranged from recovery levels of over 50 % to 74.2 %, while inhibition of BuChE by compounds 18, 19, 31, 32, 55, and 56 ranged from 48.2 % to 79.3 %. These findings underscore the potential of azinethiacrown ethers and acyclic thiacrown ethers as promising candidates for further exploration in the context of AChE and BuChE inhibition. In silico molecular docking using the CDocker tool confirmed the mechanism of action of synthetic compound 16 which has been successfully docked into the binding pocket of AChE protein with the highest score of 42.36 in comparison with the rest of the compounds.

Inhibitory effect of galantamine and donepezil combination against cholinesterase: An in silico and in vitro study.

This study aimed to evaluate the in silico and in vitro inhibitory effect of the combined use of galantamine (GAL) and donepezil (DON) against acetylcholinesterase and butyrylcholinesterase (BuChE) enzymes. In silico and in vitro cholinesterase analysis were carried out for GAL and DON alone and combined. Molecular modeling studies were carried out (docking analysis, molecular dynamics simulation, and quantum theory of atoms in molecules). Cholinesterase's inhibitory activities by modified Ellman's method and the drug combination effect using the Chou-Talalay method were assayed. GAL/DON combination showed the co-occupancy of the ligands in both enzymes through in silico studies. Regarding in vitro BuChE inhibition analyses, three of five combinations showed an interaction between GAL and DON at the threshold of additive affect (0.9 < CI < 1.1), with a tendency toward a synergistic effect for higher concentrations. This is the first report showing the efficacy of the GAL/DON combinations inhibiting BuChE, showing the importance of analyzing the behavior of different ligands when co-occupancy into the active site is possible. These combinations might be a possible therapy to improved efficacy, reduced doses, minor side effects, and high levels of the neurotransmitter in the synaptic space for Alzheimer's disease.

Subcritical water extraction of red cabbage regarding cholinesterase and tyrosinase inhibitory activity

Pressurized high-temperature processing of water where polarity can be regulated has been an advanced extraction technology considering sustainable management and efficient use of natural resources. In this study, subcritical water extraction of red cabbage and its enzyme inhibitory potential were investigated using response surface methodology. Spectrophotometric quantification was utilized for total phenol, flavonoid, and anthocyanin contents, while dominant anthocyanin molecules were identified with UPLC-DAD, ESI-MS/MS. The 200 bar-120°C-30 min-30 solvent:solid ratio extract was determined with the highest total anthocyanin content (2.33 mg Cya3GluE/g), where cyanidin-3-sophoroside-5-glucoside, cyanidin-3-(sinapoyl)-sophoroside-5-glucoside, and cyanidin-3-(sinapoyl)(sinapoyl)-sophoroside-5-glucoside were identified as the dominant anthocyanins (1.15 mg Cya3GluE/g). IC50 values for acetylcholinesterase, butyrylcholinesterase, and tyrosinase inhibitory activities were found to be 0.058 mg/ml, 0.215 mg/ml, and 0.86 mg/ml, respectively (tenfold superior to solvent extract). Despite the well-known high-temperature lability of phenolic compounds, subcritical water proved to be useful as a more selective system for extracting red cabbage bioactives with enzyme inhibitory activity when appropriate process conditions could be determined.

Experimental and pharmacoinformatic approaches unveil the neuropharmacological and analgesic potential of chloroform fraction of Roktoshirinchi (Achyranthes ferruginea Roxb.)

Ethnopharmacological relevanceAchyranthes ferruginea (A. ferruginea) Roxb. is a common plant used in traditional medicine in Asia and Africa. It has a variety of local names, including “Gulmanci” in Nigeria, “Dangar” in Pakistan, “Thola” in Ethiopia, and “Roktoshirinchi” in Bangladesh. It is edible and has several ethnomedical uses for a wide range of illnesses, including hysteria, dropsy, constipation, piles, boils, asthma, and shigellosis. However, the neuropharmacological and analgesic potential of A. ferruginea remains uninvestigated. Aim of the studyTo assess the neuropharmacological and analgesic potential of A. ferruginea through a multifaceted approach encompassing both experimental and computational models. Materials and methodsMethanol was used to extract the leaves of A. ferruginea. It was then fractionated with low to high polar solvents (n-hexane, chloroform, ethyl acetate, and water) to get different fractions, including chloroform fraction (CLF). The study selected CLF at different doses and conducted advanced chemical element and proximate analyses, as well as phytochemical profiling using GC-MS. Toxicological studies were done at 300 μg per rat per day for 14 days. Cholinesterase inhibitory potential was checked using an in-vitro colorimetric assay. Acetic acid-induced writhing (AAWT) and formalin-induced licking tests (FILT) were used to assess anti-nociceptive effects. The forced swim test (FST), tail suspension test (TST), elevated plus maze (EPM), hole board test (HBT), and light and dark box test (LDB) were among the behavioral tests used to assess depression and anxiolytic activity. Network pharmacology-based analysis was performed on selected compounds using the search tool for interacting chemicals-5 (STITCH 5), Swiss target prediction tool, and search tool for the retrieval of interacting genes and proteins (STRING) database to link their role with genes involved in neurological disorders through gene ontology and reactome analysis. ResultsQualitative chemical element analysis revealed the presence of 15 elements, including Na, K, Ca, Mg, P, and Zn. The moisture content, ash value, and organic matter were found to be 11.12, 11.03, and 88.97%, respectively. GC-MS data revealed that the CLF possesses 25 phytoconstituents. Toxicological studies suggested the CLF has no effects on normal growth, hematological and biochemical parameters, or cellular organs after 14 days at 300 μg per rat. The CLF markedly reduced the activity of both acetylcholinesterase and butyrylcholinesterase (IC50: 56.22 and 13.22 μg/mL, respectively). Promising dose-dependent analgesic activity (p < 0.05) was observed in chemically-induced pain models. The TST and FST showed a dose-dependent substantial reduction in immobility time due to the CLF. Treatment with CLF notably increased the number of open arm entries and time spent in the EPM test at doses of 200 and 400 mg/kg b.w. The CLF showed significant anxiolytic activity at 200 mg/kg b.w. in the HBT test, whereas a similar activity was observed at 400 mg/kg b.w. in the EPM test. A notable increase in the amount of time spent in the light compartment was observed in the LDB test by mice treated with CLF, suggesting an anxiolytic effect. A network pharmacology study demonstrated the relationship between the phytochemicals and a number of targets, such as PPARA, PPARG, CHRM1, and HTR2, which are connected to the shown bioactivities. ConclusionsThis study demonstrated the safety of A. ferruginea and its efficacy in attenuating cholinesterase inhibitory activity, central and peripheral pain, anxiety, and depression, warranting further exploration of its therapeutic potential.

Benzofuran Derivatives from Cortex Mori Radicis and Their Cholinesterase-Inhibitory Activity.

The phytochemical investigation of Cortex Mori Radicis led to the isolation and identification of a new prenylated benzofuranone (1) and four ring-opening derivatives (2-5) named albaphenol A-E, as well as nigranol A (6), together with ten 2-arylbenzofuran derivatives (7-16). The characterization of the structures of the new compounds and the structural revision of nigranol A (6) were conducted using the comprehensive analysis of spectroscopic data (1D/2D NMR, HRESIMS, CD, and XRD). Compounds 1-16 were tested for their inhibitory effects on acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Compounds 1 and 4 showed weak BChE-inhibitory activity (IC50 45.5 and 61.0 μM); six 2-arylbenzofuran derivatives showed more-potent BChE-inhibitory activity (IC50 2.5-32.8 μM) than the positive control galantamine (IC50 35.3 μM), while being inactive or weakly inhibitory toward AChE. Cathafuran C (14) exhibited the most potent and selective inhibitory activity against BChE in a competitive manner, with a Ki value of 1.7 μM. The structure-activity relationships of the benzofuran-type stilbenes were discussed. Furthermore, molecular docking and dynamic simulations were performed to clarify the interactions of the inhibitor-enzyme complex.