Structure-based virtual screening of the Enamine database of 1.7 million compounds followed by WaterMap calculations (a molecular dynamics simulation-based method) was applied to identify novel AChE inhibitors. The inhibitory potency of 29 selected compounds against electric eel (ee) AChE was determined using the Ellman’s method. Three compounds were found active (success rate 10%).
For the most potent compound (~40% of inhibition at 10 μM), 20 derivatives were discovered based on the Enamine similarity search. Finally, five compounds were found promising (IC 50 ranged from 6.3 µM to 17.5 µM) inhibitors of AChE. The performed similarity and fragment analysis confirmed significant structural novelty of novel AChE inhibitors. Toxicity/safety of selected compounds was determined in zebrafish model.
Palicourea tomentosa (Aubl.) Borhidi: Microscopy, chemical composition and the analgesic, anti-inflammatory and anti-acetylcholinesterase potential
Ethnopharmacological relevance: Palicourea tomentosa (Aubl.) Borhidi (synonym Psychotria poeppigiana Müll. Arg.) leaves are used in the popular treatments of inflammation and pain; however, there are no scientific studies demonstrating their activity as the methanolic extract of P. tomentosa.
Aim of study: This study was undertaken to investigate the potential antioxidant, anti-acetylcholinesterase, anti-hyperalgesic, anti-nociceptive and anti-inflammatory properties, as well as the chemical composition and concentrations of constituents of the methanolic extract of P. tomentosa leaves (MEPT). The study also analyzes the micromorphology and histochemistry of leaves of P. tomentosa.
Materials and methods: The MEPT was analysed by ultra-high-pressure liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS/MS). The concentrations of total phenols, flavonoids, flavonols and condensed tannin were determined. The micromorphology and histochemistry of leaves were performed using standard reagents, light and field emission scanning electron microscopy, beyond energy-dispersive X-ray spectroscopy. The antioxidant activity was evaluated for DPPH, β-carotene and MDA. The anti-inflammatory activity of MEPT (30, 100, and 300 mg/kg) was assayed in carrageenan-induced models of paw oedema, mechanical hyperalgesia (Von Frey), cold allodynia (acetone) and pleurisy in mice. The anti-nociceptive potential of MEPT (30, 100, and 300 mg/kg) was evaluated by the formalin method in mice. The anti-acetylcholinesterase properties were evaluated in vivo in four rat brain structures.
Results: The total ion chromatogram of MEPT demonstrated two alkaloids, one coumarin, one iridoid and two terpene derivatives. The highest phenol, flavonoid, flavonol and condensed tannin concentrations were found in the extract. A comprehensive explanation of the leaf micromorphology and histochemistry was presented. MEPT was significantly inhibited by the DPPH, β-carotene and MDA models. MEPT (30, 100 and 300 mg/kg) reduced the inflammation and hyperalgesic parameters in a carrageenan model and reduced formalin-induced nociception in both phases, which were cold sensitivity and oedema formation. The oral administration of 30 and 100 mg/kg MEPT significantly inhibited AChE activity in the frontal cortex.
Conclusion: This is the first chemical and biological study performed with a P. tomentosa methanolic extract and anatomical and histochemical analysis. The present study showed that MEPT inhibited pain and inflammatory parameters contributing, at least in part, to explain the popular use of this plant as analgesic natural agent. Also, anatomical and histochemistry of leaves described in the present study provide microscopical information, which aids species identification.
Regulation of acetylcholinesterase during the lipopolysaccharide-induced inflammatory responses in microglial cells
The non-classical function of acetylcholine (ACh) has been reported in neuroinflammation that represents the modulating factor in immune responses via activation of α7 nicotinic acetylcholine receptor (α7 nAChR), i.e., a cholinergic anti-inflammatory pathway (CAP). Acetylcholinesterase (AChE), an enzyme for ACh hydrolysis, has been proposed to have a non-classical function in immune cells. However, the involvement of AChE in neuroinflammation is unclear. Here, cultured BV2 cell, a microglial cell line, and primary microglia from rats were treated with lipopolysaccharide (LPS) to induce inflammation and to explore the regulation of AChE during this process. The expression profiles of AChE, α7 nAChR, and choline acetyltransferase (ChAT) were revealed in BV2 cells.
The expression of AChE (G4 form) was induced significantly in LPS-treated BV2 cells: the induction was triggered by NF-κB and cAMP signaling. Moreover, ACh or α7 nAChR agonist suppressed the LPS-induced production of pro-inflammatory cytokines, as well as the phagocytosis of microglia, by activating α7 nAChR and followed by the regulation of NF-κB and CREB signaling. The ACh-induced suppression of inflammation was abolished in AChE overexpressed cells, but did not show a significant change in AChE mutant (enzymatic activity knockout) transfected cells. These results indicate that the neuroinflammation-regulated function of AChE may be mediated by controlling the ACh level in the brain system.
Nanobodies as binding-chaperones stabilize the recombinant Bombyx mori acetylcholinesterase and protect the enzyme activity in pesticide detection
In our previous study, the recombinant type II acetylcholinesterase from Bombyx mori (rBmAChE) presented outstanding sensitivity to pesticides, which exhibited great potential in pesticides detection. However, the poor stability of rBmAChE and also the unclear mechanism of its sensitivity hindered the applications in on-site testing of pesticides residues. In this study, we constructed an immune nanobody library, in which we obtained 48 rBmAChE-specific nanobodies. Among them, Nb4 and Nb9 were verified as the most prominent enhancers of the enzyme activity and stabilizers under thermal stress, which indicated their usage as protective reagents for rBmAChE.
The simultaneously addition of the two Nbs enhanced the thermal-stability of rBmAChE against exposure to 50-70 °C, and also remained 100% residual activity after 30 days storage at – 20 °C or 4 °C, whereas 80% and 62% at – 80 °C and 25 °C. The homologous modeling and docking of Nb4 and Nb9 to rBmAChE indicated the stabilization of Nb4 to the peripheral anion site (PAS) of rBmAChE while Nb9 protected the C-terminal structure. Substrate docking demonstrated the importance of electrostatic attraction during catalytic process, that might be enhanced by Nbs. As a result, Nb4 and Nb9 were proved to have great potential on rBmAChE applications due to their regulation on enzyme activity and protection against thermal-inactivation and long-term storage of rBmAChE.
Acetylcholinesterase |
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| AP78858 | SAB | 1mg | 2640 EUR |
Acetylcholinesterase |
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| AP80050 | SAB | 1mg | 2640 EUR |
Acetylcholinesterase |
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| AP80187 | SAB | 1mg | 2640 EUR |
Acetylcholinesterase |
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| AP80192 | SAB | 1mg | 2640 EUR |
Acetylcholinesterase |
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| AP80202 | SAB | 1mg | 2640 EUR |
Acetylcholinesterase (AChE) |
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| 018-39 | PHOENIX PEPTIDE | 100 μg | 140.4 EUR |
Acetylcholinesterase Reagent |
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| DACEN25ML | BioAssay Systems | 100 | 89 EUR |
Acetylcholinesterase, NT (ACHE) |
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| MBS6012768-005mg | MyBiosource | 0.05(mg | 745 EUR |
Acetylcholinesterase, NT (ACHE) |
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| MBS6012768-5x005mg | MyBiosource | 5x0.05mg | 3195 EUR |
Acetylcholinesterase Antibody |
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| E38PA1007 | EnoGene | 100ul | 225 EUR |
Acetylcholinesterase, Antibody |
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| GWB-4F28A5 | GenWay Biotech | 0.1 ml | Ask for price |
Acetylcholinesterase Antibody |
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| GWB-51EE94 | GenWay Biotech | 1 ml | Ask for price |
Acetylcholinesterase, Antibody |
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| GWB-520BDB | GenWay Biotech | 0.1 mg | Ask for price |
Acetylcholinesterase Antibody |
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| GWB-7EB934 | GenWay Biotech | 0.1 ml | Ask for price |
Acetylcholinesterase Antibody |
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| R35265-100UG | NSJ Bioreagents | 100 ug | 339.15 EUR |
Acetylcholinesterase Antibody |
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| MBS9232450-01mL | MyBiosource | 0.1mL | 415 EUR |
Acetylcholinesterase Antibody |
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| MBS9232450-5x01mL | MyBiosource | 5x0.1mL | 1841 EUR |
Acetylcholinesterase Antibody |
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| MBS8580273-01mL | MyBiosource | 0.1mL | 305 EUR |
Acetylcholinesterase Antibody |
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| MBS8580273-01mLAF405L | MyBiosource | 0.1mL(AF405L) | 465 EUR |
Acetylcholinesterase Antibody |
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| MBS8580273-01mLAF405S | MyBiosource | 0.1mL(AF405S) | 465 EUR |
Acetylcholinesterase Antibody |
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| MBS8580273-01mLAF610 | MyBiosource | 0.1mL(AF610) | 465 EUR |
Acetylcholinesterase Antibody |
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| MBS8580273-01mLAF635 | MyBiosource | 0.1mL(AF635) | 465 EUR |
Acetylcholinesterase Assay Kit |
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| abx298834-100Assays | Abbexa | 100 Assays | 529.2 EUR |
Acetylcholinesterase Mouse mAb |
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| E2600521 | EnoGene | 100ul | 225 EUR |
Acetylcholinesterase Assay Kit |
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| Z5030044 | Biochain | 100 assays | 841 EUR |
Acetylcholinesterase assay kit |
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| BC096-50T24S | ELK Biotech | 50T/24S | 110 EUR |
Anti-human Glioma Cancer Potentials of Neobavaisoflavone as Natural Antioxidant Compound and Its Inhibition Profiles for Acetylcholinesterase and Butyrylcholinesterase Enzymes with Molecular Modeling and Spin Density Distributions Studies
- In this study, the carcinogenic potential of Neobavaisoflavone as a natural antioxidant compound and the inhibitory profiles of acetylcholinesterase and butyrylcholinesterase were investigated by molecular modeling and spin density distribution studies. To evaluate the antioxidant properties of neobavaisoflavone, DPPH test was performed in the presence of butyl hydroxytoluene as a control. Neobavaisoflavone cell viability was low compared to normal human glioma cancer cell lines, namely LN-229, U-87 and A-172 cell lines, without any effect of cytotoxicity on normal cell line. Neobavaisoflavone inhibited half of DPPH at 125 μg/mL.
- The best effects of Neobavaisoflavone antihypertensive glioma against the above cell lines were in the LN-229 cell line. In addition, the significant anti-cancer potential of human glioma Neobavaisoflavone against the popular human glioma cancer cell lines is related in this study. IC50 values were calculated by Neobavaisoflavone diagrams, 63.87 nM for AChE and 112.98 nM for BuChE, % Activity- [Inhibitor]. According to the above results, Neobavaisoflavone can be used to treat a variety of human glioma cancers in humans.
- In addition, molecular modeling calculations were performed to compare the biochemical activities of the Neobavaisoflavone molecule with enzymes. After molecular insertion calculations, ADME/T analysis was performed to investigate the properties of the neobavaisoflavone molecule, which will be used as a drug in the future. Then, different parameters for the antioxidant activity of the neobavaisoflavone molecule were calculated.