Patent ID: 7939248

Claim:
A method for producing a metabolite selected from the group consisting of 4-OH-tolbutamide, 4-OH-warfarin, 6-OH-warfarin, 7-OH-warfarin, 6-OH-chlorzoxazone, dextrorphan, 3-methoxy-morphinan, 3-hydroxy-morphinan, 7-HC sulfate, 7-HC glucuronide, 1-OH-midazolam, 4-OHmidazolam, and acetaminophen from an exogenous precursor, which metabolite is one that is produced by normal mammalian hepatocytes in vivo when the precursor is supplied, said method, comprising: (a) preparing cultured hepatocytes by (i) culturing a single cell suspension of mammalian hepatocytes in a culture medium for a period of between about 12 and about 168 hours in a culture vessel having a flat surface under conditions that permit aggregation of hepatocytes into spheroids which comprise viable hepatocytes; (ii) transferring said spheroids into a rotating wall vessel (RV W) culture chamber where said spheroids remain substantially in suspension during subsequent culture with rotation; (iii) incubating said suspended spheroids under conditions of vessel rotation such that said spheroids remain substantially in suspension; wherein suspended spheroids remain viable and capable of: (1) producing and secreting normal hepatocyte products; and (2) metabolizing drugs or toxins via enzymes of the cytochrome P450 system, which drugs or toxins are ones that are normally metabolized by hepatocytes in vivo for more than 30 days; (b) providing to said metabolite the precursor which is to be converted into said metabolite for a period of time sufficient to convert the precursor into said metabolite; and (c) obtaining said metabolite from the culture medium, the cells or both, thereby producing said metabolite.