Oxidation of Salicylic Acid and Aspirin by Human Cytochromes P450

Abstract

Aspirin (acetylsalicylic acid) is a commonly used analgesic. It is rapidly deacetylated both enzymatically and non‐enzymatically to salicylic acid. Salicylic acid can then be further metabolized by conjugation with glycine or glucuronides or by hydroxylation to form 2,3‐dihydroxybenzoic acid (2,3‐DHBA) and 2,5‐dihydroxybenzoic acid (2,5‐DHBA). This oxidation has been suspected to occur from oxygen radicals via Fenton reactions as well as by cytochrome P450 enzymes. We analyzed the formation of 2,3‐DHBA and 2,5‐DHBA in the presence of human liver microsomes by HPLC with fluorescence detection. We were able to determine that microsomal oxidation was much faster for salicylic acid than aspirin. 2,3‐DHBA was found to be the major oxidative product formed from salicylic acid, whereas aspirin could detect only 2,5‐DHBA. Recombinant human P450s 2C8, 2C9, 2C19, 2D6, 2E1, and 3A4 all catalyzed the 5‐hydroxylation of salicylic acid. Inhibitor studies were able to confirm that all six of these enzymes played a role in the formation of 2,3‐ and 2,5‐DHBA, as well as found that 2A6 and 2B6 played a role in the hydroxylation to 2,5‐DHBA. It was also shown in the inhibitor studies that P450 2E1 was the major enzyme contributing to the in both 3‐ and 5‐hydroxylations.This work was supported in part by National Institutes of Health grant R37 CA090426.

Pharmacokinetic Parameters of Watermelon (Rind, Flesh, and Seeds) Bioactive Components in Human Plasma: A Pilot Study to Investigate the Relationship to Endothelial Function

“…In contrast, 2,5-dihydroxybenzoic acid showed significantly higher plasma concentrations after WS intake compared to WF and WR consumption. The plasma concentration of 2,5-dihydroxybenzoic acid started to increase from 0.5 h after watermelon seed intake and reached maximum concentration at 8 h. One hypothesis for this pharmacokinetic pattern is that 2,5-dihydroxybenzoic acid could be a metabolite of other compounds, possibly salicylic acid . Salicylic acid was detected in WF, WR, and WS as free acid and in the form of salicylic acid- O -hexoside.…”