Zosuquidar (LY335979) is a widely used experimental P-glycoprotein (P-gp) inhibitor, which is commended as very potent but also as very specific for P-gp. In this in vitro and in silico study, we demonstrated for the first time that zosuquidar also inhibits organic cation transporters (OCT) 1-3, albeit less potently than P-gp. This still has to be kept in mind when zosuquidar is used to inhibit cellular efflux of P-gp substrates that are concurrently transported into the cells by OCTs. To avoid interference in these assays, zosuquidar concentrations should be kept below 1 μM.
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
- dizocilpinmaleát * farmakologie MeSH
- hematoencefalická bariéra metabolismus účinky léků MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- molekulární struktura MeSH
- neuroprotektivní látky * farmakologie chemie chemická syntéza MeSH
- potkani Sprague-Dawley MeSH
- receptory N-methyl-D-aspartátu * antagonisté a inhibitory metabolismus MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
N-methyl-D-aspartate receptors (NMDARs) play a significant role in developing several central nervous system (CNS) disorders. Currently, memantine, used for treating Alzheimer's disease, and ketamine, known for its anesthetic and antidepressant properties, are two clinically used NMDAR open-channel blockers. However, despite extensive research into NMDAR modulators, many have shown either harmful side effects or inadequate effectiveness. For instance, dizocilpine (MK-801) is recognized for its powerful psychomimetic effects due to its high-affinity and nearly irreversible inhibition of the GluN1/GluN2 NMDAR subtypes. Unlike ketamine, memantine and MK-801 also act through a unique, low-affinity "membrane-to-channel inhibition" (MCI). We aimed to develop an open-channel blocker based on MK-801 with distinct inhibitory characteristics from memantine and MK-801. Our novel compound, K2060, demonstrated effective voltage-dependent inhibition in the micromolar range at key NMDAR subtypes, GluN1/GluN2A and GluN1/GluN2B, even in the presence of Mg2+. K2060 showed reversible inhibitory dynamics and a partially trapping open-channel blocking mechanism with a significantly stronger MCI than memantine. Using hippocampal slices, 30 μM K2060 inhibited excitatory postsynaptic currents in CA1 hippocampal neurons by ∼51 %, outperforming 30 μM memantine (∼21 % inhibition). K2060 exhibited No Observed Adverse Effect Level (NOAEL) of 15 mg/kg upon intraperitoneal administration in mice. Administering K2060 at a 10 mg/kg dosage resulted in brain concentrations of approximately 2 μM, with peak concentrations (Tmax) achieved within 15 minutes. Finally, applying K2060 with trimedoxime and atropine in mice exposed to tabun improved treatment outcomes. These results underscore K2060's potential as a therapeutic agent for CNS disorders linked to NMDAR dysfunction.
- MeSH
- antagonisté excitačních aminokyselin farmakologie MeSH
- dizocilpinmaleát * farmakologie MeSH
- excitační postsynaptické potenciály účinky léků MeSH
- hipokampus účinky léků metabolismus MeSH
- lidé MeSH
- memantin farmakologie MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- neurony účinky léků metabolismus MeSH
- receptory N-methyl-D-aspartátu * antagonisté a inhibitory metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Therapeutic options for Alzheimer's disease are limited. Dual compounds targeting two pathways concurrently may enable enhanced effect. The study focuses on tacrine derivatives inhibiting acetylcholinesterase (AChE) and simultaneously N-methyl-D-aspartate (NMDA) receptors. Compounds with balanced inhibitory potencies for the target proteins (K1578 and K1599) or increased potency for AChE (K1592 and K1594) were studied to identify the most promising pro-cognitive compound. Their effects were studied in cholinergic (scopolamine-induced) and glutamatergic (MK-801-induced) rat models of cognitive deficits in the Morris water maze. Moreover, the impacts on locomotion in the open field and AChE activity in relevant brain structures were investigated. The effect of the most promising compound on NMDA receptors was explored by in vitro electrophysiology. The cholinergic antagonist scopolamine induced a deficit in memory acquisition, however, it was unaffected by the compounds, and a deficit in reversal learning that was alleviated by K1578 and K1599. K1578 and K1599 significantly inhibited AChE in the striatum, potentially explaining the behavioral observations. The glutamatergic antagonist dizocilpine (MK-801) induced a deficit in memory acquisition, which was alleviated by K1599. K1599 also mitigated the MK-801-induced hyperlocomotion in the open field. In vitro patch-clamp corroborated the K1599-associated NMDA receptor inhibitory effect. K1599 emerged as the most promising compound, demonstrating pro-cognitive efficacy in both models, consistent with intended dual effect. We conclude that tacrine has the potential for development of derivatives with dual in vivo effects. Our findings contributed to the elucidation of the structural and functional properties of tacrine derivatives associated with optimal in vivo pro-cognitive efficacy.
- MeSH
- acetylcholinesterasa metabolismus MeSH
- antagonisté excitačních aminokyselin farmakologie MeSH
- bludiště - učení * účinky léků MeSH
- cholinesterasové inhibitory * farmakologie MeSH
- dizocilpinmaleát * farmakologie MeSH
- kognice * účinky léků MeSH
- krysa rodu rattus MeSH
- paměť účinky léků MeSH
- potkani Wistar * MeSH
- receptory N-methyl-D-aspartátu * antagonisté a inhibitory metabolismus MeSH
- skopolamin MeSH
- takrin * farmakologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Targeting Anaplastic lymphoma kinase (ALK) is a promising therapeutic strategy for aberrant ALK-expressing malignancies including neuroblastoma, but resistance to ALK tyrosine kinase inhibitors (ALK TKI) is a distinct possibility necessitating drug combination therapeutic approaches. Using high-throughput, genome-wide CRISPR-Cas9 knockout screens, we identify miR-1304-5p loss as a desensitizer to ALK TKIs in aberrant ALK-expressing neuroblastoma; inhibition of miR-1304-5p decreases, while mimics of this miRNA increase the sensitivity of neuroblastoma cells to ALK TKIs. We show that miR-1304-5p targets NRAS, decreasing cell viability via induction of apoptosis. It follows that the farnesyltransferase inhibitor (FTI) lonafarnib in addition to ALK TKIs act synergistically in neuroblastoma, inducing apoptosis in vitro. In particular, on combined treatment of neuroblastoma patient derived xenografts with an FTI and an ALK TKI complete regression of tumour growth is observed although tumours rapidly regrow on cessation of therapy. Overall, our data suggests that combined use of ALK TKIs and FTIs, constitutes a therapeutic approach to treat high risk neuroblastoma although prolonged therapy is likely required to prevent relapse.
- MeSH
- anaplastická lymfomová kináza * genetika metabolismus antagonisté a inhibitory MeSH
- apoptóza účinky léků genetika MeSH
- chemorezistence genetika účinky léků MeSH
- dibenzocyklohepteny * MeSH
- farnesyltranstransferasa * antagonisté a inhibitory metabolismus MeSH
- GTP-fosfohydrolasy * genetika metabolismus MeSH
- inhibitory proteinkinas * farmakologie terapeutické užití MeSH
- lidé MeSH
- membránové proteiny metabolismus genetika MeSH
- mikro RNA * genetika metabolismus MeSH
- mutace MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- neuroblastom * farmakoterapie genetika patologie metabolismus MeSH
- piperidiny * farmakologie terapeutické užití MeSH
- pyridiny * farmakologie terapeutické užití MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- synergismus léků MeSH
- xenogenní modely - testy protinádorové aktivity MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Bolesti hlavy spojené s onemocněním COVID-19 jsou novinkou posledních čtyř let. Postcovidová bolest hlavy se může se projevit migrenózním charakterem bolesti nebo tenzní cephaleou. Bolest hlavy může přetrvávat až 60 dní a zejména z tohoto důvodu je zapotřebí věnovat diagnostice a léčbě postcovidové bolesti hlavy pozornost. Z patofyziologického hlediska je přetrvávající aktivace imunitního systému a trigeminovaskulární aktivace. Prevence by měla být zvážena pro přetrvávající bolesti hlavy a indikace analgetik má být zvolena podle fenotypu postcovidové bolesti hlavy. Léčba postcovidové bolesti hlavy do značné míry řídí stávajícími pokyny pro primární bolesti hlavy s odpovídajícím fenotypem.
Headaches associated with COVID-19 have been a new development for the past four years. Post-covid headache may present with a migrainous character of pain or tension cephalea. The headache may persist for up to 60 days and, particularly for this reason, attention should be paid to the diagnosis and treatment of post-covid headache. Pathophysiologically, there is persistent immune system activation and trigeminovascular activation. Prevention should be considered for persistent headache and the indication of analgesics should be chosen according to the phenotype of postcovid headache. Treatment of post-covid headache largely follows existing guidelines for primary headache with an appropriate phenotype.
- MeSH
- amitriptylin terapeutické užití MeSH
- bolesti hlavy * epidemiologie farmakoterapie patofyziologie MeSH
- látky ovlivňující centrální nervový systém terapeutické užití MeSH
- lidé MeSH
- neopioidní analgetika terapeutické užití MeSH
- postakutní syndrom COVID-19 * farmakoterapie patofyziologie patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- kazuistiky MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Dopamine type 2 receptors (D2Rs) constitute the main molecular target in the pharmacotherapy of schizophrenia. However, the second and third generation of antipsychotics comprises multi-target ligands, also binding serotonin type 3 receptors (5-HT3Rs) and other receptor classes as well. Here, we examined two experimental compounds (marked compound K1697 and K1700) from the group of 1,4-di-substituted aromatic piperazines, previously described in the study of Juza et al., 2021, and compared them with the chosen reference antipsychotic, aripiprazole. Their efficacy against schizophrenia-like behavior was tested in two different models of psychosis in the rat, induced by acute administration of either amphetamine (1.5 mg/kg) or dizocilpine (0.1 mg/kg), reflecting the dopaminergic and glutamatergic hypotheses of schizophrenia. The two models exhibited broadly similar behavioral manifestations: hyperlocomotion, disrupted social behavior and impaired prepulse inhibition of the startle response. However, they differed in their treatment responses as hyperlocomotion and prepulse inhibition deficit in the dizocilpine model were resistant to antipsychotic treatment, unlike the amphetamine model. One of the experimental compounds, K1700, ameliorated all the observed schizophrenia-like behaviors in the amphetamine model with an efficacy comparable to or greater than aripiprazole. Whereas social impairments caused by dizocilpine were strongly suppressed by aripiprazole, K1700 was less efficient. Taken together, K1700 showed antipsychotic properties comparable to those of aripiprazole, although the efficacy of the two drugs differed in specific domains of behavior and was also model-dependent. Our present results highlight the differences in these two schizophrenia models and their responsiveness to pharmacotherapy, and confirm compound K1700 as a promising drug candidate.
- MeSH
- amfetamin MeSH
- antipsychotika * terapeutické užití MeSH
- aripiprazol MeSH
- chinolony * MeSH
- dizocilpinmaleát MeSH
- dopamin metabolismus MeSH
- krysa rodu rattus MeSH
- psychotické poruchy * farmakoterapie MeSH
- receptory serotoninové MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Schizophrenia research has increased in recent decades and focused more on its neural basis. Decision-making and cognitive flexibility are the main cognitive functions that are impaired and considered schizophrenia endophenotypes. Cognitive impairment was recently connected with altered functions of N-methyl-d-aspartate (NMDAR) glutamatergic receptors, which increased cortical activity. Selective NMDAR antagonists, such as MK-801, have been used to model cognitive inflexibility in schizophrenia. Decreased GABAergic inhibitory activity has been shown elsewhere with enhanced cortical activity. This imbalance in the excitatory/inhibitory may reduce the entrainment of prefrontal gamma and hippocampal theta rhythms and result in gamma/theta band de-synchronization. The current study established an acute MK-801 administration model of schizophrenia-like cognitive inflexibility in rats and used the attentional set-shifting task in which rats learned to switch/reverse the relevant rule. During the task, we used in vivo optogenetic stimulations of parvalbumin-positive interneurons at specific light pulses in the prefrontal cortex and ventral hippocampus. The first experiments showed that acute dizocilpine in rats produced schizophrenia-like cognitive inflexibility. The second set of experiments demonstrated that specific optogenetic stimulation at specific frequencies of parvalbumin-positive interneurons in the prefrontal cortex and ventral hippocampus rescued the cognitive flexibility rats that received acute MK-801. These findings advance our knowledge of the pivotal role of parvalbumin interneurons in schizophrenia-like cognitive impairment and may guide further research on this severe psychiatric disorder.
- MeSH
- dizocilpinmaleát * farmakologie MeSH
- hipokampus metabolismus MeSH
- interneurony metabolismus MeSH
- kognice MeSH
- krysa rodu rattus MeSH
- optogenetika MeSH
- parvalbuminy metabolismus MeSH
- prefrontální mozková kůra metabolismus MeSH
- receptory N-methyl-D-aspartátu metabolismus MeSH
- schizofrenie * MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Schizophrenia is severe neuropsychiatric disease, which is commonly accompanied not only by positive or negative symptoms, but also by cognitive impairment. To study neuronal mechanisms underlying cognitive distortions and mechanisms underlying schizophrenia, animal pharmacological models of cognitive symptoms are commonly used. Between various cognitive impairments in schizophrenia patients, disturbed time perception has often been reported. Here, we examined temporal and spatial cognition in a modified Carousel maze task in the animal model of schizophrenia induced by non-competitive NMDA-receptor antagonists MK-801. Male Long-Evans rats (n = 18) first learned to avoid the aversive sector on a rotating arena in both dark and light intervals. We verified that during dark, rats used temporal cues, while during light they relied predominantly on spatial cues. We demonstrated that the timing strategy depends on the stable rotation speed of the arena and on the repositioning clues such as aversive stimuli. During testing (both in light and dark intervals), half of the rats received MK-801 and the control half received saline solution. We observed dose-dependent disruptions of both temporal and spatial cognition. Namely, both doses of MK-801 (0.1 and 0.12 mg/kg) significantly impaired timing strategy in the dark and increased locomotor activity. MK-801 dose 0.1 mg/kg, but not 0.12, also impaired spatial avoidance strategy in light. We found that the timing strategy is more sensitive to NMDA antagonist MK-801 than the spatial strategy. To conclude, a modified version of the Carousel maze is a useful and sensitive tool for detecting timing impairments in the MK-801 induced rodent model of schizophrenia.
- MeSH
- antagonisté excitačních aminokyselin aplikace a dávkování farmakologie MeSH
- bludiště - učení účinky léků MeSH
- chování zvířat účinky léků MeSH
- dizocilpinmaleát aplikace a dávkování farmakologie MeSH
- kognitivní dysfunkce chemicky indukované patofyziologie MeSH
- krysa rodu rattus MeSH
- modely nemocí na zvířatech MeSH
- potkani Long-Evans MeSH
- schizofrenie chemicky indukované MeSH
- učení vyhýbat se účinky léků MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
UV-induced fingerprint spectroscopy (UV-IFS), a new tool in a toolbox of analytical methods, is a powerful technique registering molecule-specific changes of fluorescence induced by UV irradiation. Analysis of fluorescence spectra of a sample prior and after UV irradiation enables an identification of a sample of a drug or pharmaceutics based on a comparison with signals of known standards. Moreover, UV-IFS uncovers the presence of undesired contaminations or intentional changes of the composition. Herein, we employ UV-IFS for qualitative as well as quantitative analysis of common medicines including analgesic/antipyretic (Acetaminophen), antihistamines (Loratadine and Desloratadine), and phosphodiesterase type 5 inhibitors (Tadalafil and Sildenafil citrate). UV irradiation (λem = 254 nm) for 2 - 10 min induced significant changes of fluorescence of the studied samples and according to the unique patterns, the quality and quantity were evaluated. Limits of detection for individual active ingredients were calculated as follows: Acetaminophen = 0.1 μg·mL-1, Loratadine = 0.1 μg·mL-1, Desloratadine = 0.01 μg·mL-1, Tadalafil = 0.04 μg·mL-1 and Sildenafil = 0.2 μg·mL-1. Moreover, genuine and fake CIALIS, VIAGRA and KAMAGRA tablets were reliably identified.
- MeSH
- loratadin * MeSH
- paracetamol * MeSH
- sildenafil citrát MeSH
- spektrální analýza MeSH
- tablety MeSH
- tadalafil MeSH
- Publikační typ
- časopisecké články MeSH
