Promising anticonvulsant and/or analgesic compounds among 5-chloro-2- or 5-chloro-4-methyl derivatives of xanthone coupled to aminoalkanol moieties-Design, synthesis and pharmacological evaluation.

A series of 10 aminoalkanol derivatives of 5-chloro-2- or 5-chloro-4-methylxanthone was synthetized and evaluated for anticonvulsant properties (MES test, mice, intraperitoneal) and compared with neurotoxicity rotarod test (NT, mice, i.p.). The best results both in terms of anticonvulsant activity and protective index value were obtained for 3: 5-chloro-2-([4-hydroxypiperidin-1-yl]methyl)-9H-xanthen-9-one hydrochloride. Compounds: 1-3, 7 and 10 revealed ED50 values in MES test: 42.78, 31.64, 25.76, 46.19 and 52.50 mg/kg b.w., respectively. 3 showed 70% and 72% of inhibition control specific binding of sigma-1 (σ1) and sigma-2 (σ2) receptor, respectively. 3 exhibited also antinociceptive activity at dose 2 mg/kg b.w. after chronic constriction injury in mice. 1, 3, 7 and 10 were evaluated on gastrointestinal flora and proved safe. In genotoxicity test (UMU-Chromotest) compounds 1, 7 and 10 proved safe at dose 150-300 µg/ml. The pharmacokinetic analysis showed rapid absorption of all studied molecules from the digestive tract (tmax  = 5-30 min). The bioavailability of the compounds ranged from 6.6% (1) to 16% (10). All studied compounds penetrate the blood-brain barrier with brain to plasma ratios varied from 4.15 (3) to 7.6 (compound 7), after i.v. administration, and from 1 (7) to 5.72 (3) after i.g. administration.

Identification of the New Metabolite of Nebivolol Using Liquid Chromatography Coupled with High-Resolution Mass Spectrometry and Chemometrics.

In this study, the phase I hepatic metabolism pathway of a cardiovascular drug nebivolol was proposed on the basis of a human liver microsomes assay with the use of LC-HR-MS coupled with the chemometric method. Six biotransformation products were found with the assistance of chemometric analysis. Five of them were identified as the previously reported products of alicyclic hydroxylation and dihydroxylation, aromatic hydroxylation, as well as alicyclic oxidation of the parent compound. Moreover, one metabolite, not reported so far, was found to be a product of N-dealkylation of nebivolol-2-amino-1-(6-fluoro-3,4-dihydro-2H-1-benzopyran-2-yl)ethan-1-ol. The novel metabolite was submitted to an in silico toxicity analysis to assess its biological properties. The applied computational methods indicated a significantly elevated risk of its mutagenic activity, compared to the parent molecule. Several metabolites of the nebivolol described in the literature were not detected in this study, indicating their non-hepatic origin.

Evaluation of the predictivity of Acute Oral Toxicity (AOT) structure-activity relationship models.

Several public efforts are aimed at discovering patterns or classifiers in the high-dimensional bioactivity space that predict tissue, organ or whole animal toxicological endpoints. The current study sought to assess and compare the predictions of the Globally Harmonized System (GHS) categories and Dangerous Goods (DG) classifications based on Lethal Dose (LD50) from several available tools (ACD/Labs, Leadscope, T.E.S.T., CATMoS, CaseUltra). External validation was done using dataset of 375 substances to demonstrate their predictive capacity. All models showed very good performance for identifying non-toxic compounds, which would be useful for DG classification, developing or triaging new chemicals, prioritizing existing chemicals for more detailed and rigorous toxicity assessments, and assessing non-active pharmaceutical intermediates. This would ultimately reduce animal use and improve risk assessments. Category-to-category prediction was not optimal, mainly due to the tendency to overpredict the outcome and the general limitations of acute oral toxicity (AOT) in vivo studies. Overprediction does not specifically pose a risk to human health, it can impact transport and material packaging requirements. Performance for compounds with LD50 ≤ 300 mg/kg (approx. 5% of the dataset) was the poorest among all groups and could be potentially improved by including expert review and read-across to similar substances.

Characterization of Phase I Hepatic Metabolites of Anti-Premature Ejaculation Drug Dapoxetine by UHPLC-ESI-Q-TOF.

Determination of the metabolism pathway of xenobiotics undergoing the hepatic pass is a crucial aspect in drug development since the presence of toxic biotransformation products may result in significant side effects during the therapy. In this study, the complete hepatic metabolism pathway of dapoxetine established according to the human liver microsome assay with the use of a high-resolution LC-MS system was described. Eleven biotransformation products of dapoxetine, including eight metabolites not reported in the literature so far, were detected and identified. N-dealkylation, hydroxylation, N-oxidation and dearylation were found to be the main metabolic reactions for the investigated xenobiotic. In silico analysis of toxicity revealed that the reaction of didesmethylation may contribute to the increased carcinogenic potential of dapoxetine metabolites. On the other hand, N-oxidation and aromatic hydroxylation biotransformation reactions possibly lead to the formation of mutagenic compounds.

Identification and characterization of citalopram new metabolites with the use of UHPLC-Q-TOF technique: In silico toxicity assessment of the identified transformation products.

In this study the metabolite profiling of citalopram with the use of human liver microsomes as well as the complementary photocatalytic method were established. This strategy allowed the detection of five metabolites of citalopram including 3-hydroxycitalopram and 3-oxocitalopram which were found as a new and not previously described metabolites of this drug The photocatalytic simulation of metabolism was carried out using tungsten (VI) oxide nanopowders with the different particle sizes, which allowed to examine the effect of this photocatalyst parameter on the mapping of metabolic processes. The accurate characterization of all observed structures was possible due to the use of ultra-high-pressure liquid chromatography and high-resolution mass spectrometry combined system as a highly useful technique in drug metabolism studies. In order to perform the toxicity prediction of citalopram and its metabolites, the acute toxicity to rodents, as well as genotoxicity, carcinogenicity, developmental toxicity and receptor-mediated toxicity was calculated basing on the in silico tools.

Mimicking of Phase I Metabolism Reactions of Molindone by HLM and Photocatalytic Methods with the Use of UHPLC-MS/MS.

Establishing the metabolism pathway of the drug undergoing the hepatic biotransformation pathway is one of the most important aspects in the preclinical discovery process since the presence of toxic or reactive metabolites may result in drug withdrawal from the market. In this study, we present the structural elucidation of six, not described yet, metabolites of an antipsychotic molecule: molindone. The elucidation of metabolites was supported with a novel photocatalytical approach with the use of WO3 and WS2 assisted photochemical reactions. An UHPLC-ESI-Q-TOF combined system was used for the registration of all obtained metabolite profiles as well as to record the high resolution fragmentation spectra of the observed transformation products. As a reference in the in vitro metabolism simulation method, the incubation with human liver microsomes was used. Chemometric comparison of the obtained profiles pointed out the use of the WO3 approach as being more convenient in the field of drug metabolism studies. Moreover, the photocatalysis was used in the direction of the main drug metabolite synthesis in order to further isolation and characterization.

Simulation of phase I metabolism reactions of selected calcium channel blockers by human liver microsomes and photochemical methods with the use of Q-TOF LC/MS.

The in vitro phase I metabolism of perhexiline and flunarizine, two calcium channel blockers was investigated during this study with the use of human liver microsomes (HLM) method compared with TiO2, WO3 and ZnO catalyzed photochemical reaction. In order to determine the structures of metabolites an quadrupole time-of-flight mass spectrometry combined with liquid chromatography (Q-TOF LC/MS) system was used. The obtained high resolution mass spectra enabled to identify thirteen products of metabolism of selected drugs including three not yet described metabolites of perhexiline and two new metabolites of flunarizine. The vast majority of metabolites were confirmed also with the participation of photocatalytic approach of the drug metabolism simulation. The comparison of all metabolic profiles made with the use of computational methods drew attention particularly to TiO2 and WO3 catalyzed photochemical reaction as similar to HLM incubation. Additionally, in silico toxicity assessment of the detected transformation products of the analyzed substances was also evaluated.

Simulation of phase I metabolism reactions of clozapine by HLM and photocatalytic methods with the use of UHPLC-ESI-MS/MS.

In this study the comparison of human liver microsomes in in vitro incubation as well as ZnO- and TiO2 -assisted photocatalytic degradation of clozapine as a mimicking method of phase I metabolism transformation was performed. Based on reversed-phase UHPLC separation and high-resolution MS/MS data, eight transformation products were identified and seven of them were found to be hepatic metabolites of the parent compound. The multivariate chemometric comparison of the obtained results shows ZnO-assisted photocatalysis to be a more suitable approach to phase I metabolism simulation. The photocatalytic experiments demonstrated that the disappearance of clozapine followed pseudo-zero order kinetics.

Imitation of phase I metabolism reactions of MAO-A inhibitors by titanium dioxide photocatalysis.

The imitation of phase I metabolism of moclobemide and toloxatone, two monoamine oxidase type A (MAO-A) inhibitors, was performed with the use of titanium dioxide photocatalytic process. Ultra high pressure liquid chromatography system coupled with an accurate hybrid ESI-Q-TOF mass spectrometer was used for the evaluation of metabolic profiles, structural elucidation of the identified transformation products and quantitative analysis of the process. Based on high resolution MS and MS/MS data, eleven transformation products were characterized in photocatalytic experiments for moclobemide and seven products for toloxatone. A significant number of these products were found as hepatic metabolites under the incubation of the selected MAO-A inhibitors with human liver microsomes (HLM). What is important, some of these HLM metabolites are not yet described in the literature. It was also found that the multivariate chemometric analysis allowed an effortless characterization of the registered metabolic profiles which can be a useful method for a fast preliminary drug metabolism study. Additionally, principal component analysis (PCA) of the registered TOF (MS) photocatalytic and HLM profiles of moclobemide and toloxatone shows that shorter irradiation time is preferred for photocatalytic metabolism experiments. A heterogeneous photocatalysis with the use of titanium dioxide was found to be a powerful tool for mimicking phase I metabolic reactions, as a fast, sensitive and inexpensive method.

Manganese neurotoxicity and protective effects of resveratrol and quercetin in preclinical research.

BACKGROUND: Exposure to Mn results in a neurological syndrome known as manganism. METHODS: We examined how 4-week Mn exposure (20mg/kg MnCl2po, 5days/week) induces neurotoxic effects in rats. Oxidized-to-reduced glutathione ratio (GSSG/GSH), malondialdehyde (MDA), superoxide dismutase (SOD) activity, catalase (CAT) activity, vitamin E content and caspase-3 activity were measured in several rat brain structures. Further, we examined protective effects of the polyphenols: resveratrol (R) or quercetin (QCT) against Mn-induced neurotoxicity. RESULTS: After exposure to Mn, we found a rise in GSSG/GSH ratio and a reduction in SOD activity in the rat striatum (STR), while in the nucleus accumbens (NAC) decreases in alpha-tocopherol content and in SOD activity were noted. In the frontal cortex (FCX), an enhancement in GSSG/GSH ratio and a reduction in SOD and CAT activities were observed. In the cerebellum (CER), a significant increase in the caspase-3 activity paralleled a rise in the GSSG/GSH ratio and a diminution of SOD activity. In the rat hippocampus (HIP), Mn evoked an enhancement in GSSG/GSH ratio. There were no changes in the MDA levels. Pretreatment with R and QCT protected against the Mn-induced (i) enhancement in GSSG/GSH ratio in the STR, (ii) decreases in the NAC alpha-tocopherol content and (iii) reduction in SOD activity in FCX, NAC and CER. CONCLUSION: Repeated Mn administration induces toxic effects in several rat brain structures and treatment with R and QCT may be a potential therapeutic strategy to attenuate the metal neurotoxicity.

Association of Calcium and Phosphate Balance, Vitamin D, PTH, and Calcitonin in Patients With Adolescent Idiopathic Scoliosis.

STUDY DESIGN: A cross-sectional study of 2 groups of patients with scoliosis, and an age-matched control group was conducted. Each of the groups such as patients with adolescent idiopathic scoliosis (AIS) as well as control group were divided additionally into 2 groups: premenarcheal and postmenarcheal girls. OBJECTIVE: The aim of the study was to determine the levels of 25-OH-vitamin D3, calcium and phosphate, parathyroid hormone (PTH), and calcitonin in serum of pre- and postmenarcheal girls with AIS and corresponding groups of scoliosis-free controls. SUMMARY OF BACKGROUND DATA: The primary etiology and pathogenesis of AIS remains unknown. It is assumed that vitamin D deficiency and genetic predisposition, for example, polymorphisms of vitamin D receptor, have a great significance. Vitamin D plays a key role in skeletal development and prevents bone atrophy, affects the absorption of calcium, maintains calcium-phosphate homeostasis, and the bone matrix mineralization. Its deficiency can result in a wide variety of skeletal deformities, low bone mass, and then leads to the disappearance of bone. Defects in trabecular bone structure and/or bone mineralization are the main features of scoliosis. Some studies have reported that Vitamin D deficiency is common among patients with AIS. The mechanism of Vitamin D action on scoliosis development is still unclear. METHODS: Determination of serum 25-OH-D3 levels was performed using high-performance liquid chromatography chromatography; concentrations of calcium and phosphate were measured using colorimetric methods, and concentration of PTH and calcitonin was measured using ELISA system. RESULTS: Reduction in the serum levels of 25-OH-D3 and calcitonin in girls with AIS compared with healthy girls was demonstrated. CONCLUSION: The phosphate-calcium balance and PTH level seem to be normal in patients with AIS. The calcitonin level in girls with AIS is 2-fold lower than in healthy subjects. It is possible that the deficiency of vitamin D can be involved in AIS. LEVEL OF EVIDENCE: 4.

Oxidative Stress in Neurodegenerative Diseases.

The pathophysiologies of neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD), and Alzheimer's disease (AD), are far from being fully explained. Oxidative stress (OS) has been proposed as one factor that plays a potential role in the pathogenesis of neurodegenerative disorders. Clinical and preclinical studies indicate that neurodegenerative diseases are characterized by higher levels of OS biomarkers and by lower levels of antioxidant defense biomarkers in the brain and peripheral tissues. In this article, we review the current knowledge regarding the involvement of OS in neurodegenerative diseases, based on clinical trials and animal studies. In addition, we analyze the effects of the drug-induced modulation of oxidative balance, and we explore pharmacotherapeutic strategies for OS reduction.

Oxidative stress as an etiological factor and a potential treatment target of psychiatric disorders. Part 1. Chemical aspects and biological sources of oxidative stress in the brain.

Oxidative stress is a dysfunctional state of living cells, caused by the disturbance of the pro-/antioxidative equilibrium. This dynamic equilibrium, constitutive for all aerobic organisms, is an inevitable necessity of maintaining the level of oxidative factors on non-destructive value to the cell. Among these factors reactive oxygen species (ROS) and reactive nitrogen species (RNS) are the best known molecules. This review article shows the current state of knowledge on the chemical specificity, relative reactivity and main sources of ROS and RNS in biological systems. As a Part 1 to the report about the role of oxidative stress in psychiatric disorders (see Smaga et al., Pharmacological Reports, this issue), special emphasis is placed on biochemical determinants in nervous tissue, which predisposed it to oxidative damage. Oxidative stress can be identified based on the analysis of various biochemical indicators showing the status of antioxidant barrier or size of the damage. In our article, we have compiled the most commonly used biomarkers of oxidative stress described in the literature with special regard to potentially effective in the early diagnosis of neurodegenerative processes.

Oxidative stress as an etiological factor and a potential treatment target of psychiatric disorders. Part 2. Depression, anxiety, schizophrenia and autism.

The pathophysiology of psychiatric diseases, including depression, anxiety, schizophrenia and autism, is far from being fully elucidated. In recent years, a potential role of the oxidative stress has been highlighted in the pathogenesis of neuropsychiatric disorders. A body of clinical and preclinical evidence indicates that psychiatric diseases are characterized by higher levels of oxidative biomarkers and with lower levels of antioxidant defense biomarkers in the brain and peripheral tissues. In this article, we review current knowledge on the role of the oxidative stress in psychiatric diseases, based on clinical trials and animal studies, in addition, we analyze the effects of drug-induced modulation of oxidative balance and explore pharmacotherapeutic strategies for oxidative stress reduction.

The impact of Triton WR-1339 induced hyperlipidemia on the effects of benzo(a)pyrene or guaiacol on α- and γ-tocopherol pools and selected markers of pro-/antioxidative balance in rat plasma and erythrocytes.

The toxicity of carcinogenic benzo(a)pyrene (BaP) can be intensified by the pro-oxidative effects of metabolic activation. The oxidatively active products can be formed during enzymatic biotransformation or in the process of co-oxygenation with lipid peroxidation. This study assesses if the acute hyperlipidemia can increase pro-oxidative effects of BaP as a factor intensifying processes of lipid peroxidation and co-oxygenation. After three days of i.p. administration of BaP or guaiacol (equimolar dose 10mg/kg b.w.) without or with the hyperlipidemia inducer-Triton WR-1339 to male Wistar rats, the levels of α- and γ-tocopherol were measured in erythrocytes and plasma together with the level of lipid peroxidation as malonyldialdehyde (MDA) concentration. Guaiacol was chosen as a reference substance due to its high ability to co-oxygenate. Additionally, the activity of superoxide dismutase (Cu,ZnSOD) in erythrocytes and plasma was monitored. In normolipaemic groups the significant decrease in erythrocyte α-tocopherol pool and the increase in lipid peroxidation level were observed after BaP or guaiacol administration. In hyperlipaemic groups, despite the increase in the level of lipid peroxidation, there were no additional effects in tocopherol pools compared to the normolipaemic groups which could be attributed to co-oxygenation. Decrease of α-tocopherol in erythrocytes was proportional to the reduction in normolipemic subjects when accounting for the migration to hyperlipemic plasma. There was no co-oxygenation effect on the activity of superoxide dismutase (Cu,ZnSOD) in blood.

Effect of diet supplemented with quinoa seeds on oxidative status in plasma and selected tissues of high fructose-fed rats.

Oxidative stress plays an important role as a mediator of damage produced by fructose metabolism. This work was designed to investigate the effect of diet supplemented with quinoa seeds on oxidative stress in plasma, heart, kidney, liver, spleen, lung, testis and pancreas of fructose administered rats. Fructose administration (310 g/kg fodder for 5 weeks) caused oxidative stress that was manifested by the increase in plasma malondialdehyde (MDA) (p<0.05), and by the non-significant changes in the enzymatic antioxidant potential in plasma and most of tissues. Co-administration of quinoa seeds (310 g/kg fodder) maintained normal activities of some enzymes. It also influenced the oxidative stress as was evidenced by decreasing MDA in plasma, and decreasing the activities of antioxidant enzymes (erythrocyte superoxide dismutase - eSOD, catalase -CAT, plasma glutathione peroxidase - pGPX). These findings demonstrate that quinoa seeds can act as a moderate protective agent against potential of fructose-induced changes in rats by reducing lipid peroxidation and by enhancing the antioxidant capacity of blood (plasma) and heart, kidney, testis, lung and pancreas.