Isobolographic analysis demonstrates the additive and synergistic effects of gemcitabine combined with fucoidan in uterine sarcomas and carcinosarcoma cells

Background: Uterine sarcomas and carcinosarcoma are associated with unfavorable prognosis. The regimens that are used in chemotherapy are associated with high incidence of side effects and usually do not significantly increase patients’ survival rates. In this study we investigated the activity and interactions between gemcitabine and fucoidan, the natural compound known for its anti-tumor properties, in human sarcomas and carcinosarcoma cell models. Methods: SK-UT-1, SK-UT1-B (carcinosarcoma), MES-SA (leiomyosarcoma), and ESS-1 (endometrial stromal sarcoma) cell lines were used for the experiments. Cells were incubated in the presence of gemcitabine, fucoidan, and mixtures, after the incubation the MTT tests were performed. In order to assess the interactions between tested compounds isobolographic analysis was performed. Additional assessments of apoptosis and cell cycle were done. Results: Additive effect of combined treatment with gemcitabine and fucoidan was observed in ESS-1 and SK-UT-1 cell line. Although the supra-additive (synergistic) effect noticed in SK-UT-1B cell line. It was not possible to determine the interactions of fucoidan and gemcitabine in MES-SA cell line due to insuffcient response to treatment. Addition of fucoidan to gemcitabine enhances its proapoptotic activity, what was observed especially in ESS-1 and SK-UT-1B cell lines. The arrest of cell cycle induced by mixture of gemcitabine and fucoidan, superior comparing gemcitabine alone was observed in SK-UT-1B. Conclusions: Obtained data showed that a combination of fucoidan and gemcitabine in uterine endometrial stromal sarcoma and carcinosarcoma cell lines has additive or even synergistic effect in decreasing cell viability. Furthermore, this drug combination induces apoptosis and arrest of cell cycle. The resistance of uterine leiomyosarcoma cell line, justifies searching for other drugs combinations to improve therapy effcacyThe research was founded by Medical University of Lublin (grants No. DS 120, DS 121 and MNmd129)

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

N-Benzyl-(2,5-dioxopyrrolidin-1-yl)propanamide (AS-1) with hybrid structure as a candidate for a broad-spectrum antiepileptic drug

In our recent studies, we identified compound N-benzyl-2-(2,5-dioxopyrrolidin-1-yl)propanamide (AS-1) as a broad-spectrum hybrid anticonvulsant which showed potent protection across the most important animal acute seizure models such as the maximal electroshock (MES) test, the subcutaneous pentylenetetrazole (s.c. PTZ) test, and the 6-Hz (32 mA) test in mice. Therefore, AS-1 may be recognized as a candidate for new anticonvulsant effective in different types of human epilepsy with a favorable safety margin profile determined in the rotarod test in mice. In the aim of further pharmacological evaluation of AS-1, in the current study, we examined its activity in the 6-Hz (44 mA) test, which is known as the model of drug-resistant epilepsy. Furthermore, we determined also the antiseizure activity in the kindling model of epilepsy induced by repeated injection of pentylenetetrazole (PTZ) in mice. As a result, AS-1 revealed relatively potent protection in the 6-Hz (44 mA) test, as well as delayed the progression of kindling induced by repeated injection of PTZ in mice at doses of 15 mg/kg, 30 mg/kg, and 60 mg/kg. Importantly, the isobolographic analysis showed that a combination of AS-1 and valproic acid (VPA) at the fixed ratio of 1:1 displayed a supra-additive (synergistic) interaction against PTZinduced seizures inmice. Thus, AS-1may be potentially used in an add-on therapy with VPA. Moreover, incubation of zebrafish larvae with AS-1 substantially decreased the number, cumulative but not the mean duration of epileptiform-like events in electroencephalographic assay. Finally, the in vitro ADME-Tox studies revealed that AS-1 is characterized by a very good permeability in the parallel artificial membrane permeability assay test, excellent metabolic stability on human liver microsomes (HLMs), no significant influence on CYP3A4/CYP2D6 activity, and moderate inhibition of CYP2C9 in a concentration of 10 $\mu$M, as well as no hepatotoxic properties in HepG2 cells (concentration of 10 $\mu$M)

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 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

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