Enzymatic and base hydrolysis studies of GEM metabolites
| Metabolite Identification3-a | Treatment Results3-b | ||
|---|---|---|---|
| β-Glucuronidase (Helix Pomatia) | β-Glucuronidase (E. coli) | Base Hydrolysis | |
| 1-O-GlcUA, 4′-OH-GEM | N.A.3-c | +3-d | + |
| 4′-O-GlcUA-GEM | −3-e | + | − |
| 1-O-GlcUA, 5′-CH2OH-GEM | − | ±3-f | + |
| 1-O-GlcUA, 5′-COOH-GEM | − | − | + |
| 5′-COO-GlcUA-GEM | − | + | + |
| 1-O-GlcUA-GEM | + | N.A. | N.A. |
Purified HPLC fractions were analyzed for their ability to be cleaved by base or by β-glucuronidase preparations
↵3-a Refer to Fig. 3 for structures of metabolites.
↵3-b Metabolites were subjected to enzyme incubations and base hydrolysis as described in Materials and Methods.
↵3-c N.A., treatment not attempted.
↵3-d +, Positive result; metabolite was cleaved.
↵3-e −, Negative result; metabolite was not cleaved.
↵3-f ±, Inconclusive result; a small percentage (less than 10%) of the metabolite was cleaved.