Preclinical evaluation of 1,2,4-triazole-based compounds targeting voltage-gated sodium channels (VGSCs) as promising anticonvulsant drug candidates

Epilepsy is a chronic neurological disorder affecting nearly 65–70 million people worldwide. Despite the observed advances in the development of new antiepileptic drugs (AEDs), still about 30-40% of patients cannot achieve a satisfactory seizure control. In our current research, we aimed at using the combined results of radioligand binding experiments, PAMPA-BBB assay and animal experimentations in order to design a group of compounds that exhibit broad spectrum of anticonvulsant activity. The synthesized 4-alkyl-5-substituted-1,2,4-triazole-3-thione derivatives were primarily screened in the maximal electroshock-induced seizure (MES) test in mice. Next, the most promising compounds (17, 22) were investigated in 6 Hz (32 mA) psychomotor seizure model. Protective effect of compound 22 was almost similar to that of levetiracetam. Moreover, these compounds did not induce genotoxic and hemolytic changes in human cells as well as they were characterized by low cellular toxicity. Taking into account the structural requirements for good anticonvulsant activity of 4-alkyl-5-aryl-1,2,4-triazole-3-thiones, it is visible that small electron-withdrawing substituents attached to phenyl ring have beneficial effects both on affinity towards VGSCs and protective activity in the animal models of epilepsy

Znaczenie kannabinoidów w funkcjonowaniu ośrodkowego układu nerwowego

At present, there is a great emphasis of public opinion on the legalisation of medical marijuana, i.e. the top parts of the cannabis plants rich in tetrahydrocannabinol (THC). Nevertheless, in the cannabis plants, there are many various cannabinoids, including cannabidiol (CBD). Scientific reports to-date indicate the possibility for using pharmacologically active cannabinoids in the treatment of such diseases/symptoms as: anorexia, vomiting, neuropathic pain, inflammatory diseases, multiple sclerosis, degenerative diseases of the central nervous system (Parkinson’s disease, Huntington’s disease, Alzheimer’s disease, Tourette’s syndrome), epilepsy, schizophrenia, and obesity. The article presents up-to-date information on the results of experimental studies concerning the effectiveness of cannabinoids, with particular consideration of diseases related with the central nervous system, including epilepsy, neuropathic pain, mental disorders, as well as obesity and anorexia.Aktualnie obserwuje się duży nacisk opinii społecznej na legalizację medycznej marihuany, czyli szczytowych części roślin konopi indyjskich bogatych w tetrahydrokannabinol (THC). Tymczasem, w konopiach jest wiele różnych kannabinoidów, między innymi kannabidiol (CBD). Aktualne doniesienia naukowe wskazują na możliwość wykorzystania farmakologicznej aktywności kannabinoidów w obszarze leczenia takich chorób/ objawów jak: anoreksja, wymioty, ból neurogenny, choroby zapalne, stwardnienie rozsiane, choroby degeneracyjne ośrodkowego układu nerwowego (choroba Parkinsona, Huntingtona, Alzheimera oraz zespół Tourette’a), padaczka, schizofrenia, otyłość. W pracy przedstawiono aktualne informacje na temat wyników prowadzonych dotychczas badań nad skutecznością kannabinoidów, ze szczególnym uwzględnieniem chorób związanych z ośrodkowym układem nerwowym, w tym: padaczką, bólem neuropatycznym, chorobami psychicznymi oraz otyłością i anoreksją

Additive interaction for three-drug combination of carbamazepine, lacosamide and lamotrigine against maximal electroshock-induced seizures – a type I isobolographic analysis

Introduction. Treatment of epilepsy patients with one antiepileptic drug often fails and then the insufficiently medicated patients need two or three antiepileptic drugs combined together to stop their seizures. However, polytherapy is always associated with drug-drug interactions whose nature may or may not be favorable for epilepsy patients. Preclinical studies on animals can help to select beneficial combinations of antiepileptic drugs that could be used in further clinical settings. Aim. To isobolographically characterize anticonvulsant effects of a combination of three antiepileptic drugs (carbamazepine, lacosamide and lamotrigine) at the fixed drug dose ratio of 1:1:1 in the mouse maximal electroshock-induced seizure test. Material and methods. Maximal electroconvulsions were evoked in male Swiss mice by a current (25 mA, 500 V, 0.2 s stimulus duration) delivered via auricular electrodes. Type I isobolographic analysis was applied to assess the interaction among carbamazepine, lacosamide and lamotrigine. Results. Isobolographic analysis revealed that the combination of carbamazepine, lacosamide and lamotrigine produced additive interaction in the mouse maximal electroshock-induced seizure test. Conclusions. Additivity among carbamazepine, lacosamide and lamotrigine in this preclinical study can be translated to clinical settings and this three-drug combination can be recommended as a treatment option for epilepsy patients who are resistant to standard treatment regimens

N-Acetyl-L-cysteine Affects Ototoxicity Evoked by Amikacin and Furosemide Either Alone or in Combination in a Mouse Model of Hearing Threshold Decrease

Drug-induced ototoxicity resulting from therapy with aminoglycoside antibiotics and loop diuretics is one of the main well-known causes of hearing loss in patients. Unfortunately, no specific protection and prevention from hearing loss are recommended for these patients. This study aimed at evaluating the ototoxic effects produced by mixtures of amikacin (AMI, an aminoglycoside antibiotic) and furosemide (FUR, a loop diuretic) in the mouse model as the hearing threshold decreased by 20% and 50% using auditory brainstem responses (ABRs). Ototoxicity was produced by the combinations of a constant dose of AMI (500 mg/kg; i.p.) on FUR-induced hearing threshold decreases, and a fixed dose of FUR (30 mg/kg; i.p.) on AMI-induced hearing threshold decreases, which were determined in two sets of experiments. Additionally, the effects of N-acetyl-L-cysteine (NAC; 500 mg/kg; i.p.) on the hearing threshold decrease of 20% and 50% were determined by means of an isobolographic transformation of interactions to detect the otoprotective action of NAC in mice. The results indicate that the influence of a constant dose of AMI on FUR-induced hearing threshold decreases was more ototoxic in experimental mice than a fixed dose of FUR on AMI-induced ototoxicity. Moreover, NAC reversed the AMI-induced, but not FUR-induced, hearing threshold decreases in this mouse model of hearing loss. NAC could be considered an otoprotectant in the prevention of hearing loss in patients receiving AMI alone and in combination with FUR

Funkcjonowanie społeczne ludzi starszych zamieszkujących obszary wiejskie

Background. The aim of this work was to assess social functioning of elderly people living in rural areas. Material and methods. The authors used own interview questionnaire to collect the data. The study was conducted in 504 citizens, older than 65 years, from eight villages. Results. Women were more frequently widowed than men, had low educational level and lived alone. As most responses showed (51.98%), the received benefits did not cover the respondents’ current needs and 5.95% of them claimed that they were insufficient. The remaining respondents, i.e. 42.06%, stated that the available resources fulfilled their needs. The respondents would also point to family’s aid (n=411; 81.55%) or spouse’s aid (n=147; 29.56%). Only 37 people benefited from social care (n=504; 7.34%), of which 24 (64.86%) claimed that the help provided by social care was insufficient. Conclusions. Old women in rural areas tend to live alone more frequently, are widowed and have lower level of education than men. The received financial benefits do not fully cover current needs of older residents of rural areas regardless of sex. Among people of over 65 years living in rural areas, the majority (81.55%) would point to family support and only 2.18% indicated social care as a source of income.Wprowadzenie. Celem pracy była ocena funkcjonowania społecznego osób w podeszłym wieku zamieszkujących obszary wiejskie. Materiał i metody. Dane w badaniu zostały zebrane przy pomocy autorskiego kwestionariusza. Badania ankietowe przeprowadzono wśród 504 obywateli ośmiu wsi u osób powyżej 65 roku życia. Wyniki. Kobiety były częściej wdowami niż mężczyźni, miały niski poziom wykształcenia i żyły samotnie. Zgodnie z większością odpowiedzi (51,98%), otrzymane świadczenia nie pokrywają bieżących potrzeb, 5,95% badanych stwierdziło, że zasoby były niewystarczające. Pozostała liczba respondentów 42,06% stwierdziła, że zasoby zaspokajają ich potrzeby. Najczęściej wskazywano na pomoc rodziny (n = 411; 81,55%) lub współmałżonka (n = 147, 29,56%). Tylko 37 osób korzystało z pomocy społecznej (n = 504; 7,34%), spośród których 24 osoby (64,86%) twierdziły, że pomoc zapewniona przez opiekę społeczną jest niewystarczająca. Wnioski. Starsze kobiety żyjące na obszarach wiejskich częściej mieszkają samotnie, są wdowami i mają niższe wykształcenie niż mężczyźni. Wszystkie otrzymywane korzyści finansowe nie w pełni pokrywają bieżące potrzeby starszych mieszkańców obszarów wiejskich bez względu na płeć. Wśród osób powyżej 65 roku życia mieszkających na obszarach wiejskich większość (81,55%) wskazała pomoc rodzinną, a 2,18% wskazało opiekę społeczną jako źródło wsparcia

Synergy, Additivity and Antagonism between Esculetin and Six Commonly Used Chemotherapeutics in Various Malignant Melanoma Cell Lines—An Isobolographic Analysis

(1) Malignant melanomas are dangerous skin cancers, and the treatment of melanomas with various cytostatic drugs often causes side effects and after their prolonged use resistance to these drugs appears. The aim of this study was to evaluate the anticancer effects of esculetin (a simple coumarin) and to assess pharmacodynamic interactions between esculetin and six commonly used cytostatic drugs (cisplatin, epirubicin, docetaxel, paclitaxel, mitoxantrone and vemurafenib) using an isobolographic analysis. (2) The experiments were carried out on four human malignant melanoma cell lines (FM55P, A375, FM55M2 and SK-MEL28). The effects of esculetin on viability, cell proliferation and cytotoxicity were verified in the range of concentrations of 2–200 μM. (3) Esculetin inhibited, in a dose-dependent manner, malignant melanoma cell viability and proliferation. The IC50 for esculetin ranged from 18.20 ± 2.93 to 120.64 ± 30.39 μM depending on the melanoma cell lines used. The combinations of esculetin with epirubicin and vemurafenib showed antagonistic interactions, the combinations of esculetin with cisplatin, docetaxel and paclitaxel showed additive interactions. For the combinations of esculetin with mitoxantrone, the isobolographic analysis displayed synergy. (4) In the treatment of malignant melanoma, esculetin should not be combined with epirubicin or vemurafenib, due to the reduction of their anticancer effects, while the synergistic interactions (esculetin + mitoxantrone) deserve a preclinical recommendation as a beneficial combination during anticancer therapy

Synergy, Additivity, and Antagonism between Cisplatin and Selected Coumarins in Human Melanoma Cells

(1) Cisplatin (CDDP) is used in melanoma chemotherapy, but it has many side effects. Hence, the search for natural substances that can reduce the dose of CDDP, and CDDP-related toxicity, is highly desired. Coumarins have many biological properties, including anticancer and antiproliferative effects. (2) An in vitro 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay on two human melanoma cell lines (FM55P and FM55M2) examined the antitumor properties of CDDP and five naturally occurring coumarins (osthole, xanthotoxin, xanthotoxol, isopimpinellin, and imperatorin). The antiproliferative effects produced by combinations of CDDP with the coumarins were assessed using type I isobolographic analysis. (3) The most potent anticancer properties of coumarins were presented by osthole and xanthotoxol. These compounds were characterized by the lowest median inhibitory concentration (IC50) values relative to the FM55P and FM55M2 melanoma cells. Isobolographic analysis showed that for both melanoma cell lines, the combination of CDDP and osthole exerted synergistic and additive interactions, while the combination of CDDP and xanthotoxol exerted additive interactions. Combinations of CDDP with xanthotoxin, isopimpinellin, and imperatorin showed antagonistic and additive interactions in two melanoma cell lines. (4) The combination of CDDP and osthole was characterized by the most desirable synergistic interaction. Isobolographic analysis allows the selection of potential candidates for cancer drugs among natural substances

Daphnetin, a Coumarin with Anticancer Potential against Human Melanoma: In Vitro Study of Its Effective Combination with Selected Cytostatic Drugs

(1) The treatment of metastatic or drug-resistant melanoma is still a significant therapeutic problem. The aim of this study was to evaluate the anticancer potential of daphnetin (7,8-dihydroxycoumarin) and its combinations with five different cytostatic drugs (mitoxantrone, docetaxel, vemurafenib, epirubicin and cisplatin). (2) The viability, proliferation and cytotoxicity of daphnetin against four human malignant melanoma cell lines were evaluated. The interactions were assessed using isobolographic analysis for the combinations of daphnetin with each of the five cytostatic drugs. (3) Daphnetin showed anticancer activity against malignant melanoma, with IC50 values ranging from 40.48 ± 10.90 µM to 183.97 ± 18.82 µM, depending on the cell line. The combination of daphnetin with either vemurafenib or epirubicin showed an antagonistic interaction. Moreover, additive interactions were observed for the combinations of daphnetin with cisplatin and docetaxel. The most desirable synergistic interactions for human melanoma metastatic cell lines were observed for the combination of daphnetin with mitoxantrone. (4) The obtained results suggest that daphnetin should not be combined with vemurafenib or epirubicin in the treatment of malignant melanoma due to the abolition of their anticancer effects. The combination of daphnetin with mitoxantrone is beneficial in the treatment of metastatic melanoma due to their synergistic interaction

Antiseizure Effects of Scoparone, Borneol and Their Impact on the Anticonvulsant Potency of Four Classic Antiseizure Medications in the Mouse MES Model—An Isobolographic Transformation

Numerous botanical drugs containing coumarins and terpenes are used in ethnomedicine all over the world for their various therapeutic properties, especially those affecting the CNS system. The treatment of epilepsy is based on antiseizure medications (ASMs), although novel strategies using naturally occurring substances with confirmed antiseizure properties are being developed nowadays. The aim of this study was to determine the anticonvulsant profiles of scoparone (a simple coumarin) and borneol (a bicyclic monoterpenoid) when administered separately and in combination, as well as their impact on the antiseizure effects of four classic ASMs (carbamazepine, phenytoin, phenobarbital and valproate) in the mouse model of maximal electroshock-induced (MES) tonic-clonic seizures. MES-induced seizures were evoked in mice receiving the respective doses of the tested natural compounds and classic ASMs (when applied alone or in combinations). Interactions for two-drug and three-drug mixtures were assessed by means of isobolographic transformation of data. Polygonograms were used to illustrate the types of interactions occurring among drugs. The total brain content of ASMs was measured in mice receiving the respective drug treatments with fluorescent polarization immunoassay. Scoparone and borneol, when administered alone, exerted anticonvulsant properties in the mouse MES model. The two-drug mixtures of scoparone with valproate, borneol with phenobarbital and borneol with valproate produced synergistic interactions in the mouse MES model, while the remaining tested two-drug mixtures produced additivity. The three-drug mixtures of scoparone + borneol with valproate and phenobarbital produced synergistic interactions in the mouse MES model. Verification of total brain concentrations of valproate and phenobarbital revealed that borneol elevated the total brain concentrations of both ASMs, while scoparone did not affect the brain content of these ASMs in mice. The synergistic interaction of scoparone with valproate observed in the mouse MES model is pharmacodynamic in nature. Borneol elevated the brain concentrations of the tested ASMs, contributing to the pharmacokinetic nature of the observed synergistic interactions with valproate and phenobarbital in the mouse MES model