Publikace se zaměřuje na farmakorezistenci u schizofrenie, deprese, úzkosti a obsedantně-kompulzivní poruchy. Určeno odborné veřejnosti.; Publikace se zaměřuje na problematiku rezistence vůči psychofarmakům u pacientů se závažnými duševními onemocněními, jako jsou schizofrenie, bipolární porucha, deprese, vybrané úzkostné stavy a obsedantně-kompulzivní porucha. Autorka přináší přehled výzkumných metod a vývoje v oblasti léčby farmakorezistentních stavů a zároveň popisuje slepé cesty a výzkumné přístupy, které se původně jevily jako slibné, ale v praxi se ukázaly jako neúčinné. Vysvětluje mechanismy působení psychofarmak a rozkrývá genetické základy farmakogenomiky, které pomáhají objasnit individuální variace v účincích léků. Odhaluje také složitost výzkumu a metodologické nástrahy, jež ovlivňují interpretaci studií na poli farmakorezistence. Kniha je cenným zdrojem informací pro odborníky, kteří se chtějí zorientovat v komplexnosti farmakorezistentních stavů a efektivněji podporovat své pacienty na cestě k zotavení.

• MeSH
• Mental Disorders MeSH
• Drug Resistance MeSH
• Psychotropic Drugs MeSH
• Publication type
• Monograph MeSH

• Conspectus
• Psychiatrie
• NML Fields
• psychiatrie
• psychofarmakologie

• MeSH
• Aggression psychology   MeSH
• Diagnosis, Differential * MeSH
• Child MeSH
• Anti-N-Methyl-D-Aspartate Receptor Encephalitis diagnosis   psychology   MeSH
• Infant MeSH
• Humans MeSH
• Adolescent MeSH
• Brain Neoplasms diagnosis   complications   MeSH
• Nervous System Diseases diagnosis   classification   psychology   MeSH
• Child Behavior Disorders * diagnosis   classification   MeSH
• Speech Disorders diagnosis   etiology   psychology   MeSH
• Child, Preschool MeSH
• Reye Syndrome chemically induced   diagnosis   MeSH
• Check Tag
• Child MeSH
• Infant MeSH
• Humans MeSH
• Adolescent MeSH
• Male MeSH
• Child, Preschool MeSH
• Female MeSH
• Publication type
• Case Reports MeSH

N-Methyl-d-aspartate receptors (NMDARs) play a crucial role in excitatory neurotransmission, with numerous pathogenic variants identified in the GluN subunits, including their ligand-binding domains (LBDs). The prevailing hypothesis postulates that the endoplasmic reticulum (ER) quality control machinery verifies the agonist occupancy of NMDARs, but this was tested in a limited number of studies. Using microscopy and electrophysiology in the human embryonic kidney 293 (HEK293) cells, we found that surface expression of GluN1/GluN2A receptors containing a set of alanine substitutions within the LBDs correlated with the measured EC50 values for glycine (GluN1 subunit mutations) while not correlating with the measured EC50 values for l-glutamate (GluN2A subunit mutations). The mutant cycle of GluN1-S688 residue, including the pathogenic GluN1-S688Y and GluN1-S688P variants, showed a correlation between relative surface expression of the GluN1/GluN2A receptors and the measured EC50 values for glycine, as well as with the calculated ΔGbinding values for glycine obtained from molecular dynamics simulations. In contrast, the mutant cycle of GluN2A-S511 residue did not show any correlation between the relative surface expression of the GluN1/GluN2A receptors and the measured EC50 values for l-glutamate or calculated ΔGbinding values for l-glutamate. Coexpression of both mutated GluN1 and GluN2A subunits led to additive or synergistic alterations in the surface number of GluN1/GluN2A receptors. The synchronized ER release by ARIAD technology confirmed the altered early trafficking of GluN1/GluN2A receptors containing the mutated LBDs. The microscopical analysis from embryonal rat hippocampal neurons (both sexes) corroborated our conclusions from the HEK293 cells.

• MeSH
• Glycine metabolism   MeSH
• HEK293 Cells MeSH
• Hippocampus cytology   metabolism   MeSH
• Rats MeSH
• Glutamic Acid metabolism   MeSH
• Humans MeSH
• Ligands MeSH
• Mutation genetics   MeSH
• Protein Domains MeSH
• Nerve Tissue Proteins MeSH
• Receptors, N-Methyl-D-Aspartate * metabolism   genetics   chemistry   MeSH
• Protein Transport physiology   genetics   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Alzheimer's disease (AD) is a multifaceted neurodegenerative disorder for which current treatments provide only symptomatic relief, primarily through cholinesterase (ChE) inhibition and N-methyl-d-aspartate receptor (NMDAR) antagonism. To improve therapeutic efficacy and safety, we designed and synthesized 16 novel tacrine derivatives modified at position 7 with various (hetero)aryl groups or deuterium substitution. Initially, in silico screening predicted favorable CNS permeability and oral bioavailability. Subsequent in vitro evaluations demonstrated significant inhibitory potency against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with derivatives 5i and 5m displaying particularly promising profiles. Metabolic stability assessed using human liver microsomes revealed enhanced stability for compound 5e, whereas 5i and 5m underwent rapid metabolism. Notably, compound 7 showed improved metabolic stability attributed to deuterium incorporation. The newly synthesized compounds were further tested for antagonistic activity on the GluN1/GluN2B subtype of NMDAR, with compound 5m exhibiting the most potent and voltage-independent inhibition. The ability of these compounds to permeate the blood-brain barrier (BBB) was confirmed through in vitro PAMPA assays. In preliminary hepatotoxicity screening (HepG2 cells), most derivatives exhibited higher cytotoxicity than tacrine, emphasizing the ongoing challenge in hepatotoxicity management. Based on its overall favorable profile, compound 5m advanced to in vivo pharmacokinetic studies in mice, demonstrating efficient CNS penetration, with brain concentrations exceeding plasma levels (brain-to-plasma ratio 2.36), indicating active transport across the BBB. These findings highlight compound 5m as a promising tacrine-based multi-target-directed ligand, supporting further preclinical development as a potential therapeutic candidate for AD.

• MeSH
• Acetylcholinesterase metabolism   MeSH
• Alzheimer Disease * drug therapy   metabolism   MeSH
• Biological Availability MeSH
• Butyrylcholinesterase metabolism   MeSH
• Cholinesterase Inhibitors * pharmacology   chemistry   chemical synthesis   MeSH
• Blood-Brain Barrier metabolism   MeSH
• Microsomes, Liver metabolism   MeSH
• Humans MeSH
• Ligands MeSH
• Molecular Structure MeSH
• Mice MeSH
• Receptors, N-Methyl-D-Aspartate * antagonists & inhibitors   metabolism   MeSH
• Tacrine * pharmacology   chemistry   chemical synthesis   MeSH
• Dose-Response Relationship, Drug MeSH
• Structure-Activity Relationship MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

The central nervous system is a well-known steroidogenic tissue producing, among others, cholesterol metabolites such as neuroactive steroids, oxysterols and steroid hormones. It is well known that these endogenous molecules affect several receptor classes, including ionotropic GABAergic and NMDA glutamatergic receptors in neurons. It has been shown that also ionotropic purinergic (P2X) receptors are cholesterol metabolites' targets. Among P2X receptors, purinergic P2X4 and P2X7 receptors are expressed in microglia, the innate immune cells involved in the brain inflammatory response. In this study, we explore the ionotropic purinergic receptors modulation by cholesterol metabolites in microglia. Patch-clamp experiments were performed in BV2 cells, a murine microglia cell line, to evaluate effects of cholesterol metabolites using micro- and nanomolar concentrations. About P2X4 receptor, we found that testosterone butyrate (20 μM and 200 nM) and allopregnanolone (10 μM and 100 nM) both potentiated its current, while neither 25-hydroxycholesterol (10 μM and 100 nM) nor 17β-estradiol (1 μM) showed any effects. On the other hand, P2X7 receptor current was potentiated by allopregnanolone (10 μM) and 25-hydroxycholesterol (10 μM and 100 nM). Taken together, our data show that modulation of either P2X4 and P2X7 current is affected differently by cholesterol metabolites, suggesting a structure-activity relationship among these players. Identifying the possible link between purinergic transmission, microglia and cholesterol metabolites will allow to define new targets for drug development to treat neuroinflammation.

• MeSH
• Cell Line MeSH
• Microglia * metabolism   MeSH
• Pregnanolone * metabolism   MeSH
• Receptors, Purinergic P2X4 * metabolism   MeSH
• Testosterone * metabolism   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH

• Publication type
• Published Erratum MeSH

We aimed to prepare novel dibenzo [a,d][7]annulen derivatives that act on N-methyl-d-aspartate (NMDA) receptors with potential neuroprotective effects. Our approach involved modifying the tropane moiety of MK-801, a potent open-channel blocker known for its psychomimetic side effects, by introducing a seven-membered ring with substituted base moieties specifically to alleviate these undesirable effects. Our in silico analyses showed that these derivatives should have high gastrointestinal absorption and cross the blood-brain barrier (BBB). Our pharmacokinetic studies in rats supported this conclusion and confirmed the ability of leading compounds 3l and 6f to penetrate the BBB. Electrophysiological experiments showed that all compounds exhibited different inhibitory activity towards the two major NMDA receptor subtypes, GluN1/GluN2A and GluN1/GluN2B. Of the selected compounds intentionally differing in the inhibitory efficacy, 6f showed high relative inhibition (∼90 % for GluN1/GluN2A), while 3l showed moderate inhibition (∼50 %). An in vivo toxicity study determined that compounds 3l and 6f were safe at 10 mg/kg doses with no adverse effects. Behavioral studies demonstrated that these compounds did not induce hyperlocomotion or impair prepulse inhibition of startle response in rats. Neuroprotective assays using a model of NMDA-induced hippocampal neurodegeneration showed that compound 3l at a concentration of 30 μM significantly reduced hippocampal damage in rats. These results suggest that these novel dibenzo [a,d][7]annulen derivatives are promising candidates for developing NMDA receptor-targeted therapies with minimal psychotomimetic side effects.

• MeSH
• Dizocilpine Maleate * pharmacology   MeSH
• Blood-Brain Barrier metabolism   drug effects   MeSH
• Rats MeSH
• Humans MeSH
• Molecular Structure MeSH
• Neuroprotective Agents * pharmacology   chemistry   chemical synthesis   MeSH
• Rats, Sprague-Dawley MeSH
• Receptors, N-Methyl-D-Aspartate * antagonists & inhibitors   metabolism   MeSH
• Dose-Response Relationship, Drug MeSH
• Structure-Activity Relationship MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Anti-N-methyl D-aspartate receptor (anti-NMDAR) encephalitis is an autoimmune disorder characterized by IgG antibodies targeting NMDAR. The prevalence is remarkably higher in women and some develop the condition during pregnancy. While immunotherapies have shown good outcomes for pregnant mothers and their infants, the impact on early neurodevelopment remains elusive. This study investigates the effects of anti-NMDAR antibody on the development of primary cortical cultures. Anti-NMDAR antibody was administered to the cultures at day in vitro 5 for the following 5 days to assess dendritic branching and arbor complexity, and at day in vitro 14 for measuring the expression of brain-derived neurotrophic factor (BDNF) and synaptic proteins. Immature cultured neurons treated with anti-NMDAR antibody exhibited impaired dendritic branching and arbor complexity. Interestingly, BDNF expression was unaffected in mature neurons. Additionally, GluN1 expression, a mandatory NMDAR subunit, was significantly reduced, while no significant alterations were observed in PSD-95, gephyrin and synaptophysin expression. These findings shed light on the structural and synaptic impacts of anti-NMDAR antibody on immature neurons, providing evidence for their consequences in early neuronal development.

• MeSH
• Dendrites * drug effects   metabolism   MeSH
• Rats MeSH
• Cells, Cultured MeSH
• Membrane Proteins metabolism   immunology   MeSH
• Brain-Derived Neurotrophic Factor * metabolism   MeSH
• Neurons * metabolism   drug effects   MeSH
• Disks Large Homolog 4 Protein metabolism   MeSH
• Nerve Tissue Proteins immunology   metabolism   MeSH
• Receptors, N-Methyl-D-Aspartate * immunology   MeSH
• Synaptophysin metabolism   MeSH
• Carrier Proteins MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Alzheimerova choroba (AD) a další neurodegenerativní onemocnění vedoucí k demenci představují závažný zdravotní, sociální a ekonomický problém projevující se jako kognitivní porucha a demence. Současná léčba poklesu kognitivních funkcí je založena na podávání inhibitorů acetylcholinesterázy (AChEI) a / nebo memantinu, antagonisty N-methyl-D-aspartátového receptoru (NMDAR). Tedy, současná potenciace cholinergního přenosu a současné blokování Ca2 + excitotoxicity se jeví jako slibná strategie přinejmenším k oddálení progrese onemocnění, zpomalení neurodegenerace a ke zlepšení kognivních funkcí. Motivováni předchozími výsledky budeme zkoumat vliv memantinu na expresi NDMAR u lidských vzorků a následně potenciální přínosy duálně působících sloučenin jakožto kognitivních stimulantů a protektivních látek proti neurogeneraci. V rámci projektu budeme aplikovat in vitro a in vivo proces vývoje léčiv včetně validace farmakodynamického účinku pomocí GMO zvířat, tak abychom vyvinuli duálně účinné klinické kandidáty; Alzheimer ́s disease (AD) and other neurodegenerative diseases leading to dementia represent a serious health, social and economic problem symptomatologically manifesting as cognitive disorder and dementia. Currently available treatment of the cognitive decline is based on the administration of acetylcholinesterase inhibitors (AChEI) and/or N-methyl-D-aspartate receptor (NMDAR) antagonist memantine. Thus, to potentiate cholinergic transmission and simultaneously block the excessive Ca2+ excitotoxicity is supposed to be a promising strategy to, at least, delay the progression of the disease and enhance cognition. Motivated by previous findings, we will investigate the effect on memantine on NDMAR expression using human samples and subsequently potential benefits of dually acting compounds as the cognitive enhancers and neuroprotectants against neurogeneration. We will perform in vitro and in vivo drug development process including the GMO animals in order to deliver dually acting clinical candidates

• Keywords
• kognice, cognition, alzheimerova choroba, Alzheimer's disease, Acetylcholinesterasa, neuroprotekce, acetylcholinesterase, Neuroprotection, duální léčiva, NMDA receptor, dual drugs, NMDA receptor,

• NML Publication type
• závěrečné zprávy o řešení grantu AZV MZ ČR

N-Methyl-d-aspartate receptors (NMDARs), encoded by GRIN genes, are ionotropic glutamate receptors playing a critical role in synaptic transmission, plasticity, and synapse development. Genome sequence analyses have identified variants in GRIN genes in patients with neurodevelopmental disorders, but the underlying disease mechanisms are not well understood. Here, we have created and evaluated a transgenic mouse line carrying a missense variant Grin2bL825V , corresponding to a de novo GRIN2B variant encoding GluN2B(L825V) found in a patient with intellectual disability (ID) and autism spectrum disorder (ASD). We used HEK293T cells expressing recombinant receptors and primary hippocampal neurons prepared from heterozygous Grin2bL825V/+ (L825V/+) and wild-type (WT) Grin2b+/+ (+/+) male and female mice to assess the functional impact of the variant. Whole-cell NMDAR currents were reduced in neurons from L825V/+ compared with +/+ mice. The peak amplitude of NMDAR-mediated evoked excitatory postsynaptic currents (NMDAR-eEPSCs) was unchanged, but NMDAR-eEPSCs in L825V/+ neurons had faster deactivation compared with +/+ neurons and were less sensitive to a GluN2B-selective antagonist ifenprodil. Together, these results suggest a decreased functional contribution of GluN2B subunits to synaptic NMDAR currents in hippocampal neurons from L825V/+ mice. The analysis of the GluN2B(L825V) subunit surface expression and synaptic localization revealed no differences compared with WT GluN2B. Behavioral testing of mice of both sexes demonstrated hypoactivity, anxiety, and impaired sensorimotor gating in the L825V/+ strain, particularly affecting males, as well as cognitive symptoms. The heterozygous L825V/+ mouse offers a clinically relevant model of GRIN2B-related ID/ASD, and our results suggest synaptic-level functional changes that may contribute to neurodevelopmental pathology.

• MeSH
• Excitatory Postsynaptic Potentials physiology   MeSH
• HEK293 Cells MeSH
• Hippocampus metabolism   MeSH
• Humans MeSH
• Mutation, Missense MeSH
• Mice, Inbred C57BL MeSH
• Mice, Transgenic * MeSH
• Mice MeSH
• Neurons metabolism   MeSH
• Neurodevelopmental Disorders * genetics   physiopathology   metabolism   MeSH
• Receptors, N-Methyl-D-Aspartate * genetics   metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH