In this study, we determined the effect of methoxime (MMB-4), asoxime (HI-6), obidoxime (LüH-6), trimedoxime (TMB-4), and pralidoxime (2-PAM) on redox homeostasis in vitro. Cultured human hepatoma cells (HepG2) were exposed to oximes at concentrations equivalent to their IC50 (assessed using MTT assay) and evaluated 1, 4 and 24 h after incubation. Additionally, intact, early and late apoptotic and necrotic cells were quantified by microcapillary flow cytometry. Intracellular levels of oxygen/nitrogen species were determined using two fluorescent probes (2',7'-dichlorodihydrofluorescein diacetate and dihydroethidium). Malondialdehyde and 3-nitrotyrosine were measured by LC-MS/MS. Non-protein thiols and non-protein disulfides were evaluated using HPLC-UV to reflect antioxidant capacity. Oxidative and nitrosative stress was induced by LüH-6, TMB-4 and MMB-4, whereas 2-PAM and HI-6 appeared as weak oxidative stressors with no activity towards nitrosative stress in HepG2 cells. Based on these results, bisquartenary oxime reactivators containing two functional oxime groups at the position 4 of pyridinium ring appear as more intense oxidative and nitrosative inducers. Activation of apoptosis and necrosis do not seem to correlate with generation of RONS. On the other hand, both processes rather reflect MDA concentrations, i.e. the damage of biomolecules.
- MeSH
- acetylcholinesterasa metabolismus MeSH
- buňky Hep G2 MeSH
- lidé MeSH
- oxidační stres účinky léků MeSH
- oximy farmakologie MeSH
- reaktivátory cholinesterasy farmakologie MeSH
- reaktivní formy dusíku metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Novel tacrine-benzyl quinolone carboxylic acid (tacrine-BQCA) hybrids were designed based on multi-target directed ligands (MTLDs) paradigm, synthesized and evaluated in vitro as inhibitors of human acetylcholinesterase (hAChE) and human butyrylcholinesterase (hBChE). Tacrine moiety is represented herein as 7-methoxytacrine, 6-chlorotacrine or unsubstituted tacrine forming three different families of seven members, i.e. 21 compounds in overall. Introducing BQCA, a positive modulator of M1 muscarinic acetylcholine receptors (mAChRs), the action of novel compounds on M1 mAChRs was evaluated via Fluo-4 NW assay on the Chinese hamster ovarian (CHO-M1WT2) cell line. All the novel tacrine-BQCA hybrids were able to block the action of hAChE and hBChE in micromolar to nanomolar range. The hAChE kinetic profile of 5p was found to be mixed-type which is consistent with our docking experiments. Moreover, selected ligands were assessed for their potential hepatotoxicity on HepG2 cell line and presumable permeation through the blood-brain barrier by PAMPA assay. Expected agonistic profile towards M1 mAChRs delivered by BQCA moiety was not confirmed. From all the hybrids, 5o can be highlighted as non-selective cholinesterase inhibitor (hAChE IC50 = 74.5 nM; hBChE IC50 = 83.3 nM) with micromolar antagonistic activity towards M1 mAChR (IC50 = 4.23 μM). A non-selective pattern of cholinesterase inhibition is likely to be valuable during the onset as well as later stages of AD.
- MeSH
- acetylcholinesterasa metabolismus MeSH
- Alzheimerova nemoc farmakoterapie metabolismus MeSH
- buněčné linie MeSH
- butyrylcholinesterasa metabolismus MeSH
- chinoliny chemie farmakologie MeSH
- cholinesterasové inhibitory chemická syntéza chemie farmakologie MeSH
- Cricetulus MeSH
- lidé MeSH
- molekulární struktura MeSH
- takrin chemie farmakologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
