Isolation and identification of seven metabolites of a water-soluble platelet aggregation inhibitor in rat urine

Abstract

1. Seven metabolites of 7-piperidino-1,2,3,4,5-tetrahydroimidazo[2,1-b]quinazolin-2-one dihydrochloride monohydrate (DN-9693) were isolated from rat urine by extraction with Amberlite XAD-2 and purification by silica gel and Sephadex LH-20 open-column chromatography and preparative high-performance liquid chromatography (hplc). The structure assignment of the metabolites was performed by field desorption mass spectrometry and 200-MHz Fourier transform nmr spectroscopic analysis and comparison with authentic standards when available.

2. DN-9693 underwent metabolism mainly at the piperidine ring to give the 4-hydroxypiperidine derivative (III) and 2-hydroxy-piperidine derivative, which is further metabolized to lactam (II) or δ-aminovaleric acid (V). The acyl side chain of V was shortened by β-oxidation to form the 3-aminopropionic acid derivative (VII). V and/or VII underwent oxidative dealkylation to give the 7-amino derivative, which was conjugated with acetic acid to form the 7-acetylamino derivative (IV). DN-9693 also underwent hydrolysis of its lactam moiety to give VI.

3. The urinary excretion of III, V and VII was determined by liquid chromatography electrochemistry (LC/EC) and V proved to be the major metabolite in rat urine.

4. A procedure is also presented for the identification of DN-9693 metabolites using LC/EC.