Identifikace peptidického konjugátu s toxickým metabolitem paracetamolu N-acetyl-p-benzochinon iminem

Abstract

Acetaminophen (paracetamol) is one of the most used analgesic drug. It is considered as a safe drug, although its administration in large doses can lead to due to known metabolism to impairment of hepatocytes with subsequent necrosis. Acetaminophen is metabolized primarily in the liver where it is converted to inactive compound by conjugation with sulphate or glucoronide. However a small proportion is metabolized by hepatic cytochrome P450 enzymes to a minor but toxic intermediate metabolite N-acetyl-p- benzoquinone imine (NAPQI). This metabolite binds with the macromolecules of the hepatic cells causing dysfunction of the enzymatic systems, structural and metabolic disarray and eventually necrotic cell death. This thesis is a part of larger work intended for preparation polyclonal antibodies which can be used for further research in vivo / in vitro. The objectives of this study are a) the confirmation that NAPQI is the toxic intermediate that can react with free thiol groups and b) the in vitro production of the formed NAPQI-peptide conjugate. Using dexamethasone-induced rat liver microsomal fraction led to the expected production of NAPQI, that was linked on a synthetic peptide containing free thiol group. This adduct was collected and purified by HPLC with gradient elution. The outcome of this...