Metabolism of Levulinate in Perfused Rat Livers and Live Rats: CONVERSION TO THE DRUG OF ABUSE 4-HYDROXYPENTANOATE

• Published in: The Journal of biological chemistry 2011-02, Vol.286 (7), p.5895-5904
• Main Authors: Harris, Stephanie R, Zhang, Guo-Fang, Sadhukhan, Sushabhan, Murphy, Anne M, Tomcik, Kristyen A, Vazquez, Edwin J, Anderson, Vernon E, Tochtrop, Gregory P, Brunengraber, Henri
• Format: Article
• Language: English
• Subjects: Animals; Calcium - pharmacology; Central Nervous System Depressants - pharmacology; Cytoplasm - enzymology; Enzyme Inhibitors - adverse effects; Enzyme Inhibitors - pharmacokinetics; Enzyme Inhibitors - pharmacology; Ethanol - pharmacology; Levulinic Acids - adverse effects; Levulinic Acids - pharmacokinetics; Levulinic Acids - pharmacology; Liver - enzymology; Male; Metabolism; Mitochondria, Liver - enzymology; Oxidation-Reduction; Pentanoic Acids - adverse effects; Pentanoic Acids - metabolism; Perfusion; Rats; Rats, Sprague-Dawley; Substance-Related Disorders
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M110.196808

Calcium levulinate (4-ketopentanoate) is used as an oral and parenteral source of calcium. We hypothesized that levulinate is converted in the liver to 4-hydroxypentanoate, a new drug of abuse, and that this conversion is accelerated by ethanol oxidation. We confirmed these hypotheses in live rats, perfused rat livers, and liver subcellular preparations. Levulinate is reduced to (R)-4-hydroxypentanoate by a cytosolic and a mitochondrial dehydrogenase, which are NADPH- and NADH-dependent, respectively. A mitochondrial dehydrogenase or racemase system also forms (S)-4-hydroxypentanoate. In livers perfused with [¹³C₅]levulinate, there was substantial CoA trapping in levulinyl-CoA, 4-hydroxypentanoyl-CoA, and 4-phosphopentanoyl-CoA. This CoA trapping was increased by ethanol, with a 6-fold increase in the concentration of 4-phosphopentanoyl-CoA. Levulinate is catabolized by 3 parallel pathways to propionyl-CoA, acetyl-CoA, and lactate. Most intermediates of the 3 pathways were identified by mass isotopomer analysis and metabolomics. The production of 4-hydroxypentanoate from levulinate and its stimulation by ethanol is a potential public health concern.