Élvezeti szerekhez történő hozzászokás in vivo és in vitro vizsgálata = In vivo and in vitro study of drugs with abuse potential

A pályázat az élvezeti szerekhez történő hozzászokás in vivo és in vitro vizsgálatát tűzte ki célul. Tanulmányoztuk a központi idegrendszerben előforuló idegi növekedési faktor és számos neuropeptid (PACAP, ghrelin, endomorfin) hatását in vivo a morfin és nikotin hozzászokáshoz, valamint in vitro körülmények között a sejtmámbránból felszabaduló eikonazoidokra. A vizsgált peptidek hatásosan módosítják az élvezeti szerekhez való hozzászokás általunk vizsgált paraméterit. Figyelmet érdemel az a tény, hogy az endogén módon előforduló endomorfin módosult származéka az anyavegyületnél biológiailag hatásosabb és mint ilyen további fejlesztés céljából potenciális vegyületként jöhet szóba. | This project is aimed at investigating the effects of drugs of abuse in vivo and in vitro. The effects of brain-derived neurotrophic factor and several neuropeptides (PACAP, ghrelin and endomorphine) were investigated on the behavioral effects of morphine and nicotine in vivo and on membrane-derived eicosanoid products in vitro. The peptides investigated can modify the behavioral response to drugs of abuse effectively under our experimental conditions. It should be emphasized that an endomorphine analogue is more efficient in biological response, than the parent molecule and it may serve as a potential candidate for future development

Neurohormonok, neuropeptidek szerepe az idegrendszer és az endokrin rendszer adaptatív működésének szabályozásában = The role of neurohormones and neuropeptides in the regulation of the adaptive function of the neural and endocrine system

Összefoglalás A vizsgált periodusban a vállalt feladat több neuropeptidnek mint a PACAP 38, orexin A és B endomorphin-1, a MERF, apelin-13, urocortin 1 , 2 és 3, valamint a ghrelinnek vizsgálata olyan idegrendszeri funkciókban mint a tanulás és memória, az állat mozgás aktivitásában uj környezetben, mint az open-field aktivitásban, és megszokott környezetben telemetriásan mérve, testhőmérséklet változásában, valamint a hypophysis-mellékvesekéreg rendszer aktivitásában. A hatások mechanizmusára vonatkozó vizsgálatokban annak kideritésére, hogy a kapott változásokban milyen transmitterek vesznek részt, az állatokat a megfelelő receptor blokkolókkal vagy antiserumokkal kezeltük és vizsgáltuk a neuropeptidek hatásában létrejövő változásokat. Az egyes neuropeptidek hyperthermiával kapcsolatos vizsgálataink során angol kutatókkal végzett kooperációban találtunk rá az isatinra, amely olyan lázcsillapitónak bizonyult, amely a prostaglandin után fejti ki hatását és amelynek egyes analogjai ezerszer erősebb lázcsillapitók mint az isatin maga. Ezek az eredmények jelenleg szabadalmaztatási eljárás alatt állnak. A részletekre vonatkozó eredmények a részletes részben találhatók. | In the reported period the aim was to find out the action of certain neuropeptides on such brain function as learning and memory, locomotion in nonfamiliar environment (open-field activity) and familiar environment, in the animals home cage, measured by telemetric measurement, body temperature and pituitary-adrenal function. The following neuropeptides were used: PACAP 38, orexin A and B, endomorphin-1, MERF, apelin-13, urocortin 1, 2 and 3 as well as ghrelin. In oder to learn the mechanism of action, the involvement of transmitters in the action of neuropeptides, different receptor blockers or antisera against peptide were utilized. During the study of the action of certain neuropeptides on body temperature, which was carried out collaboration with British partners, we discovered that isatin has strong antipyretic action and the action is located after the prostaglandin synthesis, because even the PG?induced higher temperature can be blocked by isatin. We have found that some analogues of isatin thousand times are more potent antipyretics than the isatin itself. These observation are under patenting procedure. The details are in the following section

Prediction of short- and medium-term efficacy of biosimilar infliximab therapy. Do trough levels/antidrug antibody levels or clinical/biochemical markers play a more important role?

K

Efficacy and safety of the biosimilar infliximab CT-P13 treatment in inflammatory bowel diseases: a prospective, multicentre, nationwide cohort

Background and Aims: Biosimilar infliximab CT-P13 is approved for all indications of the originator product in Europe. Prospective data on its efficacy, safety, and immunogenicity in inflammatory bowel diseases are lacking. Methods: A prospective, nationwide, multicentre, observational cohort was designed to examine the efficacy, safety, and immunogenicity of CT-P13 infliximab biosimilar in the induction treatment of Crohn’s disease [CD] and ulcerative colitis [UC]. Demographic data were collected and a harmonised monitoring strategy was applied. Early clinical remission, response, and early biochemical response were evaluated at Week 14, steroid-free clinical remission was evaluated at Week 30. Therapeutic drug level was monitored using a conventional enzyme-linked immunosorbent assay. Results: In all, 210 consecutive inflammatory bowel disease [126 CD and 84 UC] patients were included in the present cohort. At Week 14, 81.4% of CD and 77.6% of UC patients showed clinical response and 53.6% of CD and 58.6% of UC patients were in clinical remission. Clinical remission rates at Week 14 were significantly higher in CD and UC patients who were infliximab naïve, compared with those with previous exposure to the originator compound [ p < 0.05]. Until Week 30, adverse events were experienced in 17.1% of all patients. Infusion reactions and infectious adverse events occurred in 6.6% and 5.7% of all patients, respectively. Conclusions: This prospective multicentre cohort shows that CT-P13 is safe and effective in the induction of clinical remission and response in both CD and UC. Patients with previous infliximab exposure exhibited decreased response rates and were more likely to develop allergic reactions

Long term efficacy, safety and immunogenicity of biosimilar infliximab after one year in a prospective nationwide cohort

K

Valproate treatment and platelet function: The role of arachidonate metabolites

Purpose: Valproate (VPA) is an extensively used drug in the therapy of epilepsies. One of the most frequently reported side effects of VPA is hemorrhagic diathesis. Some authors emphasized the decreased platelet count as the basis of VPA-induced hemorrhagic diathesis, but some reports suggested that a significant proportion of patients with normal platelet count may still have an altered platelet function. The mechanism of the VPA-induced platelet dysfunction has not yet been elucidated. A determining element of platelet functions is the arachidonate cascade. Present ex vivo experiments were designed to determine whether a relation exists between the incidence of hemostasis caused by VPA and the effect of this drug on the arachidonate cascade of platelets. Methods: Platelets were isolated from patients receiving long-term VPA treatment (serum level, 36.04 +/- 16.12 mu g/ml; n = 10) or carbamazepine (CBZ) treatment (serum level, 5.24 +/- 2.67 mu g/ml; n = 10) and were labeled with [C-14]arachidonic acid. (CBZ-treated patients were chosen as a control group, because CBZ causes blood dyscrasias similar to those elicited by VPA, but there has been no report that CBZ induces a platelet dysfunction.) The C-14-eicosanoids were separated by means of overpressure thin-layer chromatography and determined quantitatively by liquid scintillation. Results: Even when the mean plasma concentration of the drug was low, VPA treatment reduced the activity of the arachidonate cascade in platelets. VPA effectively inhibited the cyclooxygenase pathway and the synthesis of the strong platelet aggregator thromboxane A(2). Conclusions: Inhibition of the platelet arachidonate cascade may contribute to the platelet-function alterations caused by VPA

The effects of valproate on the arachidonic acid metabolism of rat brain microvessels and of platelets

Long-term administration of the antiepileptic drug valproate can induce hematologic, hepatic and endocrine abnormalities and morphologic alterations in the brain capillaries and glial cells. Valproate elicits bone marrow suppression, reducing the number of red blood cells and platelets, and causes platelet functional abnormalities. Various data suggest that more than one mechanism of valproate-associated toxicity may exist, but the pathomechanism of cell function alterations elicited by valproate has not yet been elucidated. The reported ex vivo experiments were designed to investigate the effects of valproate on the arachidonic acid cascade of rat brain capillaries and platelets. Valproate was administered (300 mg/kg body weight/day) in the drinking water to male Wistar rats for 2 weeks. isolated platelets and brain microvessels were labelled with [C-14]arachidonic acid and the released [C-14]eicosanoids were separated by overpressure thin-layer chromatography and determined quantitatively by liquid scintillation counting. Valproate treatment reduced the synthesis of cyclooxygenase and lipoxygenase products in rat platelets, in brain microvessels valproate stimulated the synthesis of Lipoxygenase metabolites and attenuated the cyclooxygenase pathway. Modifications of the arachidonate cascade in platelets and brain microvessels may contribute to the cell function alterations caused by valproate. (C) 2000 Elsevier Science B.V. All rights reserved

Effects of 3-aminopyridine-induced seizures on platelet eicosanoid synthesis

We investigated the influence of recurrent epileptic seizures on the arachidonic acid (AA) cascade in platelets and brain microvessels, using [C-14]AA as a tracer substrate and chromatographic determination. The recurrent epileptic seizures of male Wistar rats were induced every second day with 3-aminopyridine (3-AP, 25 mg/kg ip) for two weeks. In the chronic 3-AP model, the earlier epileptic insults resulted in a decreased incidence of limbic seizures and higher survival rate at later administration of 3-AP. After 3-AP treatment, the formation of lipoxygenase products was unchanged, but the total amount of cyclooxygenase (COX) metabolites was decreased both in platelets and brain microvessels:- The reduction in COX-mediated eicosanoid synthesis after recurrent seizures was due to the decreased synthesis of vasodilator and vasoconstrictor COX metabolites. In platelets, the 3-AP-treatment reduced the synthesis of vasodilator prostacyclin (PGI(2)), prostaglandin E-2 (PGE(2)) and 12-L-hydroxy5,8, 10-heptadecatrienoic acid (12-HHT), while the synthesis of prostaglandin D-2 (PGD(2)) remained unchanged. In isolated brain capillaries, the PGD2, PGE2 and 12-HHT synthesis was decreased after recurrent seizures. As for the vasoconstrictor COX metabolites, both platelets and brain microvessels synthesized significantly lesser amount of prostaglandin F-2 alpha, (PGF(2 alpha)) and thromboxane A(2) (TxA(2)) upon 3-AP administration. Our results indicate that platelets and isolated brain capillaries synthesize significantly lesser amount of COX metabolites after chronic 3-AP treatment. The decreased conversion of AA into different COX products may play a role in the neuroprotective/preconditional adaptation of the brain against subsequent seizures

Prediction of short- And medium-term efficacy of biosimilar infliximab therapy. Do trough levels and antidrug antibody levels or clinical and biochemical markers play the more important role?

Background and Aims: Biosimilar infliximab CT-P13 received European Medicines Agency [EMA] approval in June 2013 for all indications of the originator product. In the present study, we aimed to evaluate the predictors of short- and medium-term clinical outcome in patients treated with the biosimilar infliximab at the participating inflammatory bowel disease [IBD] centres in Hungary. Methods: Demographic data were collected and a harmonised monitoring strategy was applied. Clinical and biochemical activities were evaluated at Weeks 14, 30, and 54. Trough level [TL] and anti-drug antibody [ADA] concentrations were measured by enzyme-linked immunosorbent assay [ELISA] [LT-005, Theradiag, France] at baseline at 14, 30 and 54 weeks and in two centres at Weeks 2 and 6. Results: A total of 291 consecutive IBD patients (184 Crohn's disease [CD] and 107 ulcerative colitis [UC]) were included. In UC, TLs at Week 2 predicted both clinical response and remission at Weeks 14 and 30 (clinical response/remission at Week 14: area under the curve [AUC] = 0.81, p < 0.001, cut-off: 11.5 μg/ml/AUC = 0.79, p < 0.001, cut-off: 15.3μg/ml; clinical response/remission at Week 30: AUC = 0.79, p = 0.002, cut-off: 11.5 μg/ml/AUC = 0.74, p = 0.006, cut-off: 14.5 μg/ml), whereas ADA positivity at Week 14 was inversely associated with clinical response at Week 30 [58.3% vs 84.8%,p = 0.04]. Previous anti-tumour necrosis factor [TNF] exposure was inversely associated with short-term clinical remission [Week 2: 18.8% vs 47.8%, p = 0.03, at Week 6: 38.9% vs 69.7%, p = 0.013, at Week 14: 37.5% vs 2.5%, p = 0.06]. In CD, TLs at Week 2 predicted short-term [Week 14 response/remission, AUCTLweek2= 0.715-0.721, p = 0.05/0.005] but not medium-term clinical efficacy. In addition, early ADA status by Week 14 [p = 0.04-0.05 for Weeks 14 and 30], early clinical response [p < 0.001 for Weeks 30/54] and normal C-reactive protein [CRP] at Week 14 [p = 0.005-0.0001] and previous anti-TNF exposure [p = 0.03-0.0001 for Weeks 14, 30, and 54] were associated with shortand medium-term clinical response and remission. Conclusions: In UC, early TLs were predictive for short- and medium-term clinical efficacy, whereas in CD, Week 2 TLs were associated only with short-term clinical outcomes