Comparison of the oxidation of carcinogenic aristolochic acid I and II by microsomal cytochromes P450 in vitro: experimental and theoretical approaches

Václav Martínek,Petr Hodek,Heinz H Schmeiser,František Bárta,Marie Stiborová,Volker M Arlt,Eva Frei

doi:10.1007/s00706-017-2014-9

Abstract

The herbal drug aristolochic acid, a natural mixture of 8-methoxy-6-nitrophenanthro[3,4-d]-1,3-dioxole-5-carboxylic acid (AAI) and 6-nitrophenanthro[3,4-d]-1,3-dioxole-5-carboxylic acid (AAII), is derived from Aristolochia species and is the cause of two nephropathies. Ingestion of aristolochic acid is associated with the development of urothelial tumors linked with aristolochic acid nephropathy and is implicated in the development of Balkan endemic nephropathy-associated urothelial tumors. The O-demethylated metabolite of AAI, 8-hydroxyaristolochic acid (AAIa), is the detoxification product of AAI generated by its oxidative metabolism. Whereas the formation of AAIa from AAI by cytochrome P450 (CYP) enzymes has been found in vitro and in vivo, this metabolite has not been found from AAII as yet. Therefore, the present study has been designed to compare the amenability of AAI and AAII to oxidation; experimental and theoretical approaches were used for such a study. In the case of experimental approaches, the enzyme (CYP)-mediated formation of AAIa from both carcinogens was investigated using CYP enzymes present in subcellular microsomal fractions and recombinant CYP enzymes. We found that in contrast to AAI, AAII is oxidized only by several CYP enzymatic systems and their efficiency is much lower for oxidation of AAII than AAI. Using the theoretical approaches, such as flexible in silico docking methods and ab initio calculations, contribution to explanation of these differences was established. Indeed, the results found by both used approaches determined the reasons why AAI is better oxidized than AAII; the key factor causing the differences in AAI and AAII oxidation is their different amenability to chemical oxidation.Graphical abstract

Highlights

Aristolochic acid (AA), the natural extract of plants of the Aristolochiaceae family, is a mixture of structurally related nitrophenanthrene carboxylic acids, with two major components aristolochic acid I (8-methoxy-6-nitrophenanthro[3,4-d]1,3-dioxole-5-carboxylic acid, AAI; Fig. 1) and aristolochicV
There is compelling evidence that human exposure to AA leads to chronic renal disease and upper urinary tract cancer known as aristolochic acid nephropathy (AAN) [14, 15], which is recognized as a global disease [9]
We evaluated which of the cytochrome P450 (CYP) can participate in the formation of AAIa from AAI and AAII in these rat hepatic microsomes (Fig. 3)

Summary

Introduction

Aristolochic acid (AA), the natural extract of plants of the Aristolochiaceae family, is a mixture of structurally related nitrophenanthrene carboxylic acids, with two major components aristolochic acid I (8-methoxy-6-nitrophenanthro[3,4-d]1,3-dioxole-5-carboxylic acid, AAI; Fig. 1) and aristolochicV. In addition to the abilities of CYP1A1 and 1A2 to reduce AAI and AAII, these enzymes are efficient in AAI oxidative detoxification to form the Odemethylated metabolite, 8-hydroxyaristolochic acid (AAIa) [33, 34, 36,37,38,39,40] (Fig. 1).

Results

Conclusion