Dextromethorphan metabolite profiles generated by in-capillary microreaction and subsequent electrophoretic separation

ZEISBERGEROVÁ, Aleš MÁDR a Zdeněk GLATZ. Dextromethorphan metabolite profiles generated by in-capillary microreaction and subsequent electrophoretic separation. In Book of Abtracts 16th Latin-American Symposium LACE 2010. první. Florianopolis Brazílie: prof. Emanuel Carrilho, 2010, s. 127-127.

Základní údaje

• Originální název: Dextromethorphan metabolite profiles generated by in-capillary microreaction and subsequent electrophoretic separation
• Název česky: Generování metabolitů dextromethorphanu pomocí mikroreakce v kapiláře
• Název anglicky: and subsequent electrophoretic separation
• Autoři: ZEISBERGEROVÁ, Aleš MÁDR a Zdeněk GLATZ.
• Vydání: první. Florianopolis Brazílie, Book of Abtracts 16th Latin-American Symposium LACE 2010, od s. 127-127, 1 s. 2010.
• Nakladatel: prof. Emanuel Carrilho

Další údaje

• Typ výsledku: Stať ve sborníku
• Utajení: není předmětem státního či obchodního tajemství
• Organizační jednotka: Přírodovědecká fakulta
• Klíčová slova česky: EMMA; cytochromy P450; metabolismus léčiv; dextromethorphan
• Klíčová slova anglicky: EMMA; Cytochromes P450; Drug Metabolism; Dextromethorphan
• Příznaky: Mezinárodní význam, Recenzováno

Anotace

The determination of metabolite profile of potential pharmaceuticals has a substantial role in the drug development phase. Different human-derived in vitro systems like liver slices, microsomes or recombinant enzyme systems are utilized for metabolite identification. Human liver microsomes represent the generally accepted in vitro system for these studies. They credibly mimic liver functions as they contain many drug-metabolizing enzymes, mainly cytochromes P450 (CYPs). Alternatively, recombinant cytochrome P450 enzymes (rCYP) are suitable for "frontline" predictive human metabolism studies. rCYP are a favorite in vitro system for their availability and ease employment in high throughput assays. They are a valuable tool in the CYP phenotyping, i.e. searching for which CYP enzyme is involved in the biotransformation of new agents. The metabolism pathway of dextromethorphan (DEX) was chosen as a model system. DEX is an antitusive component of many medications which is chemically a non-narcotic synthetic analog of codeine. DEX is a probe drug of CYP2D6 enzyme. CYP 2D6 is one of the most important enzymes catalyzing biotransformation of xenobiotics. Further, CYP 3A4, the most abundant CYP enzyme also contributes to DEX metabolism. Thus DEX metabolite profile should be completely contained when microsomes are applied. Nowadays, most of drug metabolism studies are performed by liquid chromatography-mass spectrometry methods characterized by off-line setup. However, capillary electrophoresis (CE) represents an innovative approach to perform automated enzyme assays. Different methods and procedures of enzymatic activity studies have been reported. The aim of our work was a simultaneous determination of the parent drug, DEX and its metabolites generated by in-capillary microreaction at direct injection of microsomes or rCYP enzymes. The analytes were separated in 50-um fused silica capillary (48 cm total length) at 14 kV. The background electrolyte consisted of tetraborate buffer (80 mM, pH 9.7) with addition of 2-propanol (8 %, v/v). The capillary temperature was kept at 37 oC and the data were recorded at 200 nm. The partial filling method enabled to combine separation and incubation buffers within one capillary thus maintaining the optimal incubation conditions. Different injection strategies will be shown and compared in the term of reaction efficacy and versatility.

Anotace česky

Byla vypracována metoda pro monitorováni reakce mikrosomů a rekombinantních formem cytochromů P450 s dextromethorphanem v kapiláře.

Anotace anglicky

The determination of metabolite profile of potential pharmaceuticals has a substantial role in the drug development phase. Different human-derived in vitro systems like liver slices, microsomes or recombinant enzyme systems are utilized for metabolite identification. Human liver microsomes represent the generally accepted in vitro system for these studies. They credibly mimic liver functions as they contain many drug-metabolizing enzymes, mainly cytochromes P450 (CYPs). Alternatively, recombinant cytochrome P450 enzymes (rCYP) are suitable for "frontline" predictive human metabolism studies. rCYP are a favorite in vitro system for their availability and ease employment in high throughput assays. They are a valuable tool in the CYP phenotyping, i.e. searching for which CYP enzyme is involved in the biotransformation of new agents. The metabolism pathway of dextromethorphan (DEX) was chosen as a model system. DEX is an antitusive component of many medications which is chemically a non-narcotic synthetic analog of codeine. DEX is a probe drug of CYP2D6 enzyme. CYP 2D6 is one of the most important enzymes catalyzing biotransformation of xenobiotics. Further, CYP 3A4, the most abundant CYP enzyme also contributes to DEX metabolism. Thus DEX metabolite profile should be completely contained when microsomes are applied. Nowadays, most of drug metabolism studies are performed by liquid chromatography-mass spectrometry methods characterized by off-line setup. However, capillary electrophoresis (CE) represents an innovative approach to perform automated enzyme assays. Different methods and procedures of enzymatic activity studies have been reported. The aim of our work was a simultaneous determination of the parent drug, DEX and its metabolites generated by in-capillary microreaction at direct injection of microsomes or rCYP enzymes. The analytes were separated in 50-um fused silica capillary (48 cm total length) at 14 kV. The background electrolyte consisted of tetraborate buffer (80 mM, pH 9.7) with addition of 2-propanol (8 %, v/v). The capillary temperature was kept at 37 oC and the data were recorded at 200 nm. The partial filling method enabled to combine separation and incubation buffers within one capillary thus maintaining the optimal incubation conditions. Different injection strategies will be shown and compared in the term of reaction efficacy and versatility.

Návaznosti

• GAP206/10/0057, projekt VaV: Název: Miniaturizovaný systém pro "on-line" studie metabolismu léčiv založený na kapilární elektroforéze

Investor: Grantová agentura ČR, Miniaturizovaný systém pro "on-line" studie metabolismu léčiv založený na kapilární elektroforéze

• LC06023, projekt VaV: Název: Integrované bioanalytické technologie pro mikroanalýzy a diagnostiku s využitím LIF a hmotnostní spektrometrie

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Integrované bioanalytické technologie pro mikroanalýzy a diagnostiku s využitím LIF a hmotnostní spektrometrie

• MSM0021622413, záměr: Název: Proteiny v metabolismu a při interakci organismů s prostředím

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Proteiny v metabolismu a při interakci organismů s prostředím