Alzheimer's disease (AD) is a complex disorder with unknown etiology. Currently, only symptomatic therapy of AD is available, comprising cholinesterase inhibitors and N-methyl-d-aspartate (NMDA) receptor antagonists. Drugs targeting only one pathological condition have generated only limited efficacy. Thus, combining two or more therapeutic interventions into one molecule is believed to provide higher benefit for the treatment of AD. In the presented study, we designed, synthesized, and biologically evaluated 15 novel fluoren-9-amine derivatives. The in silico prediction suggested both the oral availability and permeation through the blood-brain barrier (BBB). An initial assessment of the biological profile included determination of the cholinesterase inhibition and NMDA receptor antagonism at the GluN1/GluN2A and GluN1/GluN2B subunits, along with a low cytotoxicity profile in the CHO-K1 cell line. Interestingly, compounds revealed a selective butyrylcholinesterase (BChE) inhibition pattern with antagonistic activity on the NMDARs. Their interaction with butyrylcholinesterase was elucidated by studying enzyme kinetics for compound 3c in tandem with the in silico docking simulation. The docking study showed the interaction of the tricyclic core of new derivatives with Trp82 within the anionic site of the enzyme in a similar way as the template drug tacrine. From the kinetic analysis, it is apparent that 3c is a competitive inhibitor of BChE.

• Klíčová slova
• Alzheimer’s disease, N-methyl-d-aspartate receptor, acetylcholinesterase, butyrylcholinesterase, fluorene, in silico, in vitro, multi-target directed ligands,
• MeSH
• Alzheimerova nemoc farmakoterapie   enzymologie   genetika   patologie   MeSH
• butyrylcholinesterasa chemie   účinky léků   genetika   MeSH
• CHO buňky MeSH
• cholinesterasové inhibitory chemie   farmakologie   MeSH
• Cricetulus MeSH
• fluoreny chemie   farmakologie   MeSH
• hematoencefalická bariéra účinky léků   MeSH
• inhibitory enzymů farmakologie   MeSH
• lidé MeSH
• počítačová simulace MeSH
• receptory N-methyl-D-aspartátu antagonisté a inhibitory   genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• butyrylcholinesterasa MeSH
• cholinesterasové inhibitory MeSH
• fluorene MeSH Prohlížeč
• fluoreny MeSH
• inhibitory enzymů MeSH
• N-methyl D-aspartate receptor subtype 2A MeSH Prohlížeč
• NR2B NMDA receptor MeSH Prohlížeč
• receptory N-methyl-D-aspartátu MeSH

N-methyl-D-aspartate receptors (NMDARs) are ionotropic glutamate receptors that play an essential role in mediating excitatory neurotransmission in the mammalian central nervous system (CNS). Functional NMDARs are tetramers composed of GluN1, GluN2A-D, and/or GluN3A-B subunits, giving rise to a wide variety of NMDAR subtypes with unique functional properties. Here, we examined the surface delivery and functional properties of NMDARs containing mutations in the glycine-binding sites in GluN1 and GluN3A subunits expressed in mammalian cell lines and primary rat hippocampal neurons. We found that the structural features of the glycine-binding sites in both GluN1 and GluN3A subunits are correlated with receptor forward trafficking to the cell surface. In addition, we found that a potentially clinically relevant mutation in the glycine-binding site of the human GluN3A subunit significantly reduces surface delivery of NMDARs. Taken together, these findings provide novel insight into how NMDARs are regulated by their glycine-binding sites and may provide important information regarding the role of NMDARs in both physiological and pathophysiological processes in the mammalian CNS.

• MeSH
• buněčná membrána metabolismus   MeSH
• Cercopithecus aethiops MeSH
• COS buňky MeSH
• glycin metabolismus   MeSH
• HEK293 buňky MeSH
• hipokampus cytologie   MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• membránové glykoproteiny chemie   metabolismus   MeSH
• mutace genetika   MeSH
• neurony metabolismus   MeSH
• podjednotky proteinů chemie   metabolismus   MeSH
• proteinové domény MeSH
• receptory N-methyl-D-aspartátu chemie   metabolismus   MeSH
• sekvence aminokyselin MeSH
• vazebná místa MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• glycin MeSH
• GRIN3A protein, human MeSH Prohlížeč
• Grin3a protein, rat MeSH Prohlížeč
• membránové glykoproteiny MeSH
• NMDA receptor A1 MeSH Prohlížeč
• podjednotky proteinů MeSH
• receptory N-methyl-D-aspartátu MeSH