SANGUINARINE IS REDUCED BY NADH THROUGH A COVALENT ADDUCT

a 2019

SANGUINARINE IS REDUCED BY NADH THROUGH A COVALENT ADDUCT

SÁNDOR, Roman; Jiří SLANINA; Adam MIDLIK; Kristýna ŠEBRLOVÁ; Lucie NOVOTNÁ et al.

Základní údaje

Originální název

SANGUINARINE IS REDUCED BY NADH THROUGH A COVALENT ADDUCT

Autoři

SÁNDOR, Roman; Jiří SLANINA; Adam MIDLIK; Kristýna ŠEBRLOVÁ; Lucie NOVOTNÁ; Petr TÁBORSKÝ; Eva TÁBORSKÁ a Ondřej PEŠ

Vydání

20. ročník Školy hmotnostní spektrometrie, 2019

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

30104 Pharmacology and pharmacy

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14110/19:00110706

Organizační jednotka

Lékařská fakulta

ISBN

978-80-88195-14-6

Klíčová slova anglicky

benzophenanthridine alkaloids; metabolism; LC MS; NADH; biotransformation; ene cyclization

Štítky

rivok

Změněno: 17. 9. 2019 16:24, doc. Mgr. Ondřej Peš, Ph.D.

Anotace

V originále

Benzo[c]phenanthridine alkaloids are a family of compounds with cytotoxic, anti-fungal and antimicrobial properties, which are heavily metabolized by the CYP 450 system. Most of the research of these compounds has been focused rather on their biological effects than their metabolism; however, it has already been found that the first step of their biotransformation is a reduction to their respective dihydro forms. Interestingly, this reaction step may proceed with NAD(P)H even when no additional enzymatic apparatus is present. We decided to study this interaction with sanguinarine (SA), as this effect has been noted but never examined any further. From the results of Matkar et al.[1], who found an unexpected compound on RP-TLC and 1D electrophoresis of NADH and SA, we hypothesized that SA forms a negatively charged, covalent adduct with NADH. We used TLC and LC-MS to identify the compound, observed at 997 Da (singly charged) and 499 Da (doubly charged). The kinetics of the equimolar mixture and the kinetics of the adduct isolated by semi-preparative chromatography was studied by the UV-Vis and MS/MS-MS spectrometry to infer the state of all reaction compounds at different time points. The kinetics of the equimolar mixture showed a decrease in concentration of SA while the concentration of the reduced form, dihydrosanguinarine, and the adduct were increasing. The kinetics of the isolated adduct was more complicated, as the adduct partially decomposed to the reactants. The results strongly supported the hypothesis that during the non-enzymatic reduction of SA by NADH, a covalent semi-stable adduct was present, and its decomposition to products was the rate-limiting step.

Návaznosti

MUNI/A/0976/2018, interní kód MU

Název: Příspěvek chemických a biochemických metodik ke studiu molekulární podstaty vybraných patologických stavů a onemocnění

Investor: Masarykova univerzita, Příspěvek chemických a biochemických metodik ke studiu molekulární podstaty vybraných patologických stavů a onemocnění, DO R. 2020_Kategorie A - Specifický výzkum - Studentské výzkumné projekty

MUNI/A/1550/2018, interní kód MU

Název: Farmakologický výzkum v oblasti farmakokinetiky, neuropsychofarmakologie a onkologie

Investor: Masarykova univerzita, Farmakologický výzkum v oblasti farmakokinetiky, neuropsychofarmakologie a onkologie, DO R. 2020_Kategorie A - Specifický výzkum - Studentské výzkumné projekty

Citovat

SÁNDOR, Roman; Jiří SLANINA; Adam MIDLIK; Kristýna ŠEBRLOVÁ; Lucie NOVOTNÁ; Petr TÁBORSKÝ; Eva TÁBORSKÁ a Ondřej PEŠ. SANGUINARINE IS REDUCED BY NADH THROUGH A COVALENT ADDUCT. In 20. ročník Školy hmotnostní spektrometrie. 2019. ISBN 978-80-88195-14-6.

@proceedings{1558176,
   author = {Sándor, Roman and Slanina, Jiří and Midlik, Adam and Šebrlová, Kristýna and Novotná, Lucie and Táborský, Petr and Táborská, Eva and Peš, Ondřej},
   booktitle = {20. ročník Školy hmotnostní spektrometrie},
   keywords = {benzophenanthridine alkaloids; metabolism; LC MS; NADH; biotransformation; ene cyclization},
   language = {eng},
   isbn = {978-80-88195-14-6},
   title = {SANGUINARINE IS REDUCED BY NADH THROUGH A COVALENT ADDUCT},
   url = {http://www.skolams.cz/},
   year = {2019}
}

TY  - CONF
ID  - 1558176
AU  - Sándor, Roman - Slanina, Jiří - Midlik, Adam - Šebrlová, Kristýna - Novotná, Lucie - Táborský, Petr - Táborská, Eva - Peš, Ondřej
PY  - 2019
TI  - SANGUINARINE IS REDUCED BY NADH THROUGH A COVALENT ADDUCT
SN  - 9788088195146
KW  - benzophenanthridine alkaloids
KW  - metabolism
KW  - LC MS
KW  - NADH
KW  - biotransformation
KW  - ene cyclization
UR  - http://www.skolams.cz/
L2  - http://www.skolams.cz/
N2  - Benzo[c]phenanthridine alkaloids are a family of compounds with cytotoxic, anti-fungal and antimicrobial properties, which are heavily metabolized by the CYP 450 system. Most of the research of these compounds has been focused rather on their biological effects than their metabolism; however, it has already been found that the first step of their biotransformation is a reduction to their respective dihydro forms. Interestingly, this reaction step may proceed with NAD(P)H even when no additional enzymatic apparatus is present. We decided to study this interaction with sanguinarine (SA), as this effect has been noted but never examined any further. From the results of Matkar et al.[1], who found an unexpected compound on RP-TLC and 1D electrophoresis of NADH and SA, we hypothesized that SA forms a negatively charged, covalent adduct with NADH. We used TLC and LC-MS to identify the compound, observed at 997 Da (singly charged) and 499 Da (doubly charged). The kinetics of the equimolar mixture and the kinetics of the adduct isolated by semi-preparative chromatography was studied by the UV-Vis and MS/MS-MS spectrometry to infer the state of all reaction compounds at different time points. The kinetics of the equimolar mixture showed a decrease in concentration of SA while the concentration of the reduced form, dihydrosanguinarine, and the adduct were increasing. The kinetics of the isolated adduct was more complicated, as the adduct partially decomposed to the reactants. The results strongly supported the hypothesis that during the non-enzymatic reduction of SA by NADH, a covalent semi-stable adduct was present, and its decomposition to products was the rate-limiting step.
ER  -

SÁNDOR, Roman; Jiří SLANINA; Adam MIDLIK; Kristýna ŠEBRLOVÁ; Lucie NOVOTNÁ; Petr TÁBORSKÝ; Eva TÁBORSKÁ a Ondřej PEŠ. SANGUINARINE IS REDUCED BY NADH THROUGH A COVALENT ADDUCT. In \textit{20. ročník Školy hmotnostní spektrometrie}. 2019. ISBN~978-80-88195-14-6.

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