Stereoselective metabolism of pentoxifylline in vitro and in vivo in humans

Abstract

Pentoxifylline increases erythrocyte flexibility, reduces blood viscosity, and inhibits platelet aggregation and is thus used in the treatment of peripheral vascular disease. It is transformed into at least seven phase I metabolites, of which two, M1 and M5, are active. The reduction of the keto group of pentoxifylline to a secondary alcohol in M1 takes place chiefly in erythrocytes, is rapidly reversible, and creates a chiral center. The aims of this study were: to develop HPLC methods to separate the enantiomers of M1, to investigate the kinetics of the reversible biotransformation of pentoxifylline to (R)- and (S)-M1 in hemolysed erythrocyte suspension, and to quantify the formation of the enantiomers of M1 (as well as M4 and M5) after intravenous and oral administration of pentoxifylline to human volunteers. (R)- and (S)-M1 could be separated preparatively on a cellobiohydrolase column, while determination in blood or plasma was by HPLC after chiral derivatization with diacetyl-L-tartaric acid anhydride. The metabolism of pentoxifylline to (R)-M1 in suspensions of hemolysed erythrocytes followed simple Michaelis-Menten kinetics (K(m) = 11 mM), while that to (S)-M1 was best described by a two-enzyme model (K(m) = 1.1 and 132 mM). Studies with inhibitors indicated that the enzymes were of the carbonyl reductase type. At a therapeutic blood concentration of pentoxifylline, the calculated rate of formation of (S)-M1 is 15 times higher than that of the (R)-enantiomer. Back-conversion of M1 to pentoxifylline was 3-4 times faster for the (S)- than for the (R)-enantiomer. In vivo, the R:S plasma concentration ratio of M1 ranged from 0.010-0.025 after intravenous infusion of 300 or 600 mg of pentoxifylline, and from 0.019-0.037 after oral administration of 600 mg. The biotransformation of pentoxifylline to M1 was thus highly stereoselective in favor of the (S)-enantiomer both in vitro and in vivo.

Human Carbonyl Reduction Pathways and a Strategy for Their Study In Vitro

“…The stereochemistry of ketone reduction has been reported for a number of xenobiotics (Asami et al, 1996;Benedetti et al, 1991Benedetti et al, , 1993Breyer-Pfaff and Nill, 1995;Dow and Berg, 1995;Eyles and Pond, 1992;Hermans and Thijssen, 1989;Kume et al, 2000;Lawrence et al, 1990;McMahon et al, 1965;Nagata et al, 1992;Nicklasson et al, 2002;Rush et al, 1990;Skálová et al, 2003;Takasaki and Tanaka, 1992;Vickers et al, 1993;Volosov et al, 1999;Weil et al, 1989;Wsól et al, 1998a). However, the absolute configuration has not been determined in all cases.…”

Pharmacokinetic-Pharmacodynamic Modeling of Methylxanthine Derivatives in Mice Challenged with High-Dose Lipopolysaccharide

“…The almost 2-fold higher value of K m observed for lisofylline may be explained by the differences in metabolic pathways of these compounds. It has been shown that in human microsomes lisofylline is metabolized by potentially saturable cytochrome P450 enzymes, such as CYP3A4, CYP1A2 and to a minor extent CYP2E1 [26,27] , whereas most of the pentoxifylline metabolism is cytochrome-P-450 independent [28] . The values of pharmacokinetic parameters of lisofylline estimated in this study are quite similar to those reported previously in healthy CD-1 mice [29] .…”

Bioanalysis of pentoxifylline and related metabolites in plasma samples through LC‐MS/MS

“…3,7-Dimethyl-1-(5′-hydroxyhexyl)xanthine (namely lisofylline or M1), 1-(3′-carboxypropyl)-3,7-dimethylxanthyne (M5) and 1-(4′-carboxybutyl)-3,7-dimethylxanthine (M4) are the metabolites frequently cited in literature (Ward and Clissold, 1987;Bryce et al, 1989;Rocci et al, 1987), resulting from in-vivo reduction (M1) or oxidation (M4, M5) processes. Other four nonconjugated metabolites are known (Nicklasson et al, 2002). PTX is reduced to M1 in the human liver.…”