Disposition and metabolism of cannabinoids.

Předkládaná dizertační práce popisuje formou komentáře k vlastním originálním publikacím výsledky výzkumu problematiky kanabinoidů, tj. fytokanabinoidů a některých syntetických kanabinoidů, jejich farmakokinetiky a účinků. Práce je složena ze čtyř tematických okruhů: farmakokinetika delta-9- tetrahydrokanabinolu (THC) a kanabidiolu (CBD) u potkanů v závislosti na způsobu podání; časový profil THC u člověka (po inhalační aplikaci) a implikace pro bezpečnost v dopravě; farmakokinetický profil syntetických kanabinoidů u potkanů; extrakce a stanovení fytokanabinoidů v rostlinném materiálu. Předmětem prvního okruhu dizertační práce bylo stanovení farmakokinetických profilů THC, CBD po jejich jednotlivém a kombinovaném podání (váhový poměr 1:1) potkanům s ohledem na podání obvyklá u člověka, a to podání inhalační, perorální a subkutánní. V průběhu studie byl sledován časový profil hladin THC a jeho metabolitů (11-hydroxy-tetrahydrokanabinol, 11-OH-THC; kyselina 11-nor-delta-9- karboxytetrahydrokanabinolová, THCOOH) a/nebo CBD v séru a mozcích zvířat ve 24 hodinovém experimentálním intervalu. Kromě inhalačního podání, po aplikaci perorální a subkutánní koadministrace CBD inhibovala metabolismus THC, což vedlo k nárůstu koncentrací THC v séru i mozku pokusných potkanů, vzhledem ke kontrolním hodnotám po...This thesis describes in the form of a commentary on own original publications research on the problems of cannabinoids, ie. phytocannabinoids and some synthetic cannabinoids, their pharmacokinetics and effects. The work consists of four thematic areas: the pharmacokinetics of delta-9- tetrahydrocannabinol (THC) and cannabidiol (CBD) in rats, depending on the route of administration; THC concentration time profile in humans (after inhalation) and implications for transport safety; the pharmacokinetic profile of synthetic cannabinoids in rats; extraction and determination of phytocannabinoids in plant material. The first part of the thesis was to determine pharmacokinetic profiles of THC, CBD and combination thereof (1:1 weight ratio) in rats with respect to administration common in humans, i.e. inhalation, oral and subcutaneous administration. THC, its metabolites (11-hydroxy-tetrahydrocannabinol, 11-OH-THC; 11-nor-delta-9- carboxytetrahydrocannabinol, THCOOH) and CBD concentrations in serum and brains of animals were monitored at the 24 hours experimental interval during the study. Except for inhalation administration, co-administration of CBD inhibited THC metabolism (after both oral and subcutaneous), resulting in an increase in THC concentrations in both serum and brain of the rats relative to...Institute of Legal Medicine and Toxicology First Faculty of Medicine Charles UniversityÚstav soudního lékařství a toxikologie 1. LF UK a VFN1. lékařská fakultaFirst Faculty of Medicin

Disposition and metabolism of cannabinoids

Předkládaná dizertační práce popisuje formou komentáře k vlastním originálním publikacím výsledky výzkumu problematiky kanabinoidů, tj. fytokanabinoidů a některých syntetických kanabinoidů, jejich farmakokinetiky a účinků. Práce je složena ze čtyř tematických okruhů: farmakokinetika delta-9- tetrahydrokanabinolu (THC) a kanabidiolu (CBD) u potkanů v závislosti na způsobu podání; časový profil THC u člověka (po inhalační aplikaci) a implikace pro bezpečnost v dopravě; farmakokinetický profil syntetických kanabinoidů u potkanů; extrakce a stanovení fytokanabinoidů v rostlinném materiálu. Předmětem prvního okruhu dizertační práce bylo stanovení farmakokinetických profilů THC, CBD po jejich jednotlivém a kombinovaném podání (váhový poměr 1:1) potkanům s ohledem na podání obvyklá u člověka, a to podání inhalační, perorální a subkutánní. V průběhu studie byl sledován časový profil hladin THC a jeho metabolitů (11-hydroxy-tetrahydrokanabinol, 11-OH-THC; kyselina 11-nor-delta-9- karboxytetrahydrokanabinolová, THCOOH) a/nebo CBD v séru a mozcích zvířat ve 24 hodinovém experimentálním intervalu. Kromě inhalačního podání, po aplikaci perorální a subkutánní koadministrace CBD inhibovala metabolismus THC, což vedlo k nárůstu koncentrací THC v séru i mozku pokusných potkanů, vzhledem ke kontrolním hodnotám po...This thesis describes in the form of a commentary on own original publications research on the problems of cannabinoids, ie. phytocannabinoids and some synthetic cannabinoids, their pharmacokinetics and effects. The work consists of four thematic areas: the pharmacokinetics of delta-9- tetrahydrocannabinol (THC) and cannabidiol (CBD) in rats, depending on the route of administration; THC concentration time profile in humans (after inhalation) and implications for transport safety; the pharmacokinetic profile of synthetic cannabinoids in rats; extraction and determination of phytocannabinoids in plant material. The first part of the thesis was to determine pharmacokinetic profiles of THC, CBD and combination thereof (1:1 weight ratio) in rats with respect to administration common in humans, i.e. inhalation, oral and subcutaneous administration. THC, its metabolites (11-hydroxy-tetrahydrocannabinol, 11-OH-THC; 11-nor-delta-9- carboxytetrahydrocannabinol, THCOOH) and CBD concentrations in serum and brains of animals were monitored at the 24 hours experimental interval during the study. Except for inhalation administration, co-administration of CBD inhibited THC metabolism (after both oral and subcutaneous), resulting in an increase in THC concentrations in both serum and brain of the rats relative to...Katedra analytické chemieDepartment of Analytical ChemistryFaculty of SciencePřírodovědecká fakult

Behavioral and Pharmacokinetic Profile of Indole-Derived Synthetic Cannabinoids JWH-073 and JWH-210 as Compared to the Phytocannabinoid Δ9-THC in Rats

Synthetic cannabinoid compounds are marketed as “legal” marijuana substitutes, even though little is known about their behavioral effects in relation to their pharmacokinetic profiles. Therefore, in the present study we assessed the behavioral effects of systemic treatment with the two synthetic cannabinoids JWH-073 and JWH-210 and the phytocannabinoid Δ9-THC on locomotor activity, anxiety-like phenotype (in the open field) and sensorimotor gating (measured as prepulse inhibition of the acoustic startle response, PPI), in relation to cannabinoid serum levels. Wistar rats were injected subcutaneously (sc.) with JWH-073 (0.1, 0.5, or 5 mg/kg), JWH-210 (0.1, 0.5, or 5 mg/kg), Δ9-THC (1 or 3 mg/kg) or vehicle (oleum helanti) in a volume of 0.5 ml/kg and tested in the open field and PPI. Although JWH-073, JWH-210, Δ9-THC (and its metabolites) were confirmed in serum, effects on sensorimotor gating were absent, and locomotor activity was only partially affected. Δ9-THC (3 mg/kg) elicited an anxiolytic-like effect as suggested by the increased time spent in the center of the open field (p < 0.05). Our results further support the potential anxiolytic-like effect of pharmacological modulation of the endocannabinoid system

Optimization of conditions for derivatization of fatty acids and perfluorinated organic acids with methylchloroformate

Statistical methods designed for optimization were applied for selection of optimal experimental conditions of derivatization process for perfluorinated acids, PFA (C8, C10 ,C12), fatty acids, FA (C8, C12, C14) and their mixtures. The calibration curves and the extraction efficiences for system acetonitrile/heptane were measured for all studied acids. The Plackett-Burman screening was performed for obtaining the significant experimental parameters from their original series and significant ones were remitted to optimization via central composite design, which takes a second order effects and mutual interactions in between parameters into account. Optimal conditions of derivatization were found for PFA, FA and their mixtures. The principle of the search of optimal selective derivatization conditions for one group of acids in the presence of the other is based on the search of optimum on the response surface where the dependent variable is either sum or difference of acid group responses normalized to common internal standard. It was found that FA influence total response more than PFA and that in the case of demand for selective derivatization of PFA in the presence of FA in the sample, the most significant parameter is the volume of methylchloroformate added. The complete selectivity of..

Preparation and testing of new highly fluorinated chloroformates used as derivatizing agents for analysis of perfluorinated acids

Katedra analytické chemieDepartment of Analytical ChemistryFaculty of SciencePřírodovědecká fakult

Disposition and metabolism of cannabinoids.

This thesis describes in the form of a commentary on own original publications research on the problems of cannabinoids, ie. phytocannabinoids and some synthetic cannabinoids, their pharmacokinetics and effects. The work consists of four thematic areas: the pharmacokinetics of delta-9- tetrahydrocannabinol (THC) and cannabidiol (CBD) in rats, depending on the route of administration; THC concentration time profile in humans (after inhalation) and implications for transport safety; the pharmacokinetic profile of synthetic cannabinoids in rats; extraction and determination of phytocannabinoids in plant material. The first part of the thesis was to determine pharmacokinetic profiles of THC, CBD and combination thereof (1:1 weight ratio) in rats with respect to administration common in humans, i.e. inhalation, oral and subcutaneous administration. THC, its metabolites (11-hydroxy-tetrahydrocannabinol, 11-OH-THC; 11-nor-delta-9- carboxytetrahydrocannabinol, THCOOH) and CBD concentrations in serum and brains of animals were monitored at the 24 hours experimental interval during the study. Except for inhalation administration, co-administration of CBD inhibited THC metabolism (after both oral and subcutaneous), resulting in an increase in THC concentrations in both serum and brain of the rats relative to..

Anticonvulsive Effects and Pharmacokinetic Profile of Cannabidiol (CBD) in the Pentylenetetrazol (PTZ) or N-Methyl-D-Aspartate (NMDA) Models of Seizures in Infantile Rats

In spite of use of cannabidiol (CBD), a non-psychoactive cannabinoid, in pediatric patients with epilepsy, preclinical studies on its effects in immature animals are very limited. In the present study we investigated anti-seizure activity of CBD (10 and 60 mg/kg administered intraperitoneally) in two models of chemically induced seizures in infantile (12-days old) rats. Seizures were induced either with pentylenetetrazol (PTZ) or N-methyl-D-aspartate (NMDA). In parallel, brain and plasma levels of CBD and possible motor adverse effects were assessed in the righting reflex and the bar holding tests. CBD was ineffective against NMDA-induced seizures, but in a dose 60 mg/kg abolished the tonic phase of PTZ-induced generalized seizures. Plasma and brain levels of CBD were determined up to 24 h after administration. Peak CBD levels in the brain (996 ± 128 and 5689 ± 150 ng/g after the 10- and 60-mg/kg doses, respectively) were reached 1–2 h after administration and were still detectable 24 h later (120 ± 12 and 904 ± 63 ng/g, respectively). None of the doses negatively affected motor performance within 1 h after administration, but CBD in both doses blocked improvement in the bar holding test with repeated exposure to this task. Taken together, anti-seizure activity of CBD in infantile animals is dose and model dependent, and at therapeutic doses CBD does not cause motor impairment. The potential risk of CBD for motor learning seen in repeated motor tests has to be further examined

Citlivá CE-MS metoda pro monitorování hladin riociguátu a desmethylriociguátu v lidském séru

Riociguat is novel antihypertensive drug for treatment of pulmonary hypertension. As such, it is still being tested in many clinical and pharmacokinetic trials. Existing methods that determine serum riociguat and desmethylriociguat (DMR) are based solely on liquid chromatography with mass spectrometry. Therefore, we present a novel capillary electrophoresis with mass spectrometry method (CE-MS) for their determination in human serum as alternative method for ongoing trials. Complete resolution of both analytes was achieved by means of pH optimization of ammonium formate background electrolytes that are fully compatible with ESI/MS detection. Simple liquid-liquid extraction was used as sample pretreatment. The calibration dependence of the method was linear (in the range of 10-1000 ng/mL), with adequate accuracy (90.1-114.9%) and precision (13.4%). LOD and LOQ were arbitrarily set at 10 ng/mL for both analytes. Clinical applicability was validated using serum samples from patients treated with riociguat in pharmacokinetic study and the results corresponded with reference HPLC-MS/MS values. Capillary electrophoresis proved to be sensitive and selective tool for the analysis of riociguat and DMR.Riociguat je nové antihypertenzivum pro léčbu plicní hypertenze. Jako takový je stále testován v mnoha klinických a farmakokinetických studiích. Stávající metody stanovení sérového riociguátu a desmethylriociguátu (DMR) jsou založeny výhradně na kapalinové chromatografii s hmotnostní spektrometrií. Tato práce prezentuje kapilární elektroforézu s metodou hmotnostní spektrometrie (CE-MS) pro jejich stanovení v lidském séru jako alternativní metodu pro probíhající studie. Úplného rozlišení obou analytů bylo dosaženo pomocí optimalizace pH elektrolytů založených na mravenčanu amonném, ​​které jsou plně kompatibilní s detekcí ESI / MS. Jako předúprava vzorku byla použita jednoduchá extrakce kapalina-kapalina. Závislost kalibrace metody byla lineární (v rozmezí 10–1000 ng/ml), s odpovídající přesností (90,1–114,9%) a precizností (13,4%). LOD a LOQ byly u obou analytů nastaveny na 10 ng/ml. Klinická použitelnost byla ověřena pomocí vzorků séra od pacientů léčených riociguatem ve farmakokinetické studii a výsledky odpovídaly referenčním hodnotám HPLC-MS/MS. Kapilární elektroforéza se ukázala jako citlivý a selektivní nástroj pro analýzu riociguátu a DMR

Dohled nad trhem: analýza parfémů na přítomnost alergenů a zakázaných látek

The market surveillance study was based on chemical analysis of 166 commercially available perfuming products in order to identify the presence of 24 regulated allergens and 21 prohibited substances. For this purpose, several GC methods were tested. The analytical approach for determination of 24 regulated allergens was based on GC-MS and GC-MS/MS analysis. Analyses were performed on two different GC columns to avoid potentially overestimated results due to matrix component co-elutions. Allergens determined by the chemical analyses were compared with the allergens declared on the product label to verify whether these products comply with the requirements of the Regulation No 1223/2009 of the European Parliament and of the Council. A specific proportion of the tested cosmetic products (43%) was found as non-compliant either due to a missing list of ingredients or due to the presence of undeclared allergens that were found to be present in the product above limits (0.001%) for required labelling. The GC-MS/MS analysis of 21 prohibited substances did not reveal any of the prohibited compounds in a concentration above LOQ except safrole, which was found in 12 out of 166 tested samples. The concentration of safrole did not exceed the concentration limit permitted by legislation for the presence from natural sources in any of these samples.Studie byla založena na chemické analýze 166 komerčně dostupných parfémovacích produktů s cílem identifikovat přítomnost 24 regulovaných alergenů a 21 zakázaných látek. Za tímto účelem bylo testováno několik metod GC. Analytický přístup pro stanovení 24 regulovaných alergenů byl založen na analýze GC-MS a GC-MS/MS. Analýzy byly provedeny na dvou různých GC kolonách, aby se zabránilo potenciálně nadhodnoceným výsledkům v důsledku společných elucí složek matrice. Alergeny stanovené chemickými rozbory byly porovnány s alergeny deklarovanými na etiketě výrobku, aby se ověřilo, zda tyto výrobky splňují požadavky Nařízení Evropského parlamentu a Rady č. 1223/2009. Konkrétní část testovaných kosmetických přípravků (43 %) byla shledána jako nevyhovující buď kvůli chybějícímu seznamu složek, nebo kvůli přítomnosti nedeklarovaných alergenů, které byly ve výrobku nalezeny nad limity (0,001 %) pro požadované označení. GC-MS/MS analýza 21 zakázaných látek neodhalila žádnou ze zakázaných sloučenin v koncentraci nad LOQ kromě safrolu, který byl nalezen ve 12 ze 166 testovaných vzorků. Koncentrace safrolu v žádném z těchto vzorků nepřekročila koncentrační limit povolený legislativou pro přítomnost z přírodních zdrojů