In Vitro Metabolism of DWP16001, a Novel Sodium-Glucose Cotransporter 2 Inhibitor, in Human and Animal Hepatocytes

Abstract

DWP16001 is currently in a phase 2 clinical trial as a novel anti-diabetes drug for the treatment of type 2 diabetes by selective inhibition of sodium-glucose cotransporter 2. This in vitro study was performed to compare the metabolism of DWP16001 in human, dog, monkey, mouse, and rat hepatocytes, and the drug-metabolizing enzymes responsible for the metabolism of DWP16001 were characterized using recombinant human cytochrome 450 (CYP) and UDP-glucuronosyltransferase (UGT) enzymes expressed from cDNAs. The hepatic extraction ratio of DWP16001 in five species ranged from 0.15 to 0.56, suggesting that DWP16001 may be subject to species-dependent and weak-to-moderate hepatic metabolism. Five phase I metabolites (M1–M5) produced by oxidation as well as three DWP16001 glucuronides (U1–U3) and two hydroxy-DWP16001 (M1) glucuronides (U4, U5), were identified from hepatocytes incubated with DWP16001 by liquid chromatography-high resolution mass spectrometry. In human hepatocytes, M1, M2, M3, U1, and U2 were identified. Formation of M1 and M2 from DWP16001 was catalyzed by CYP3A4 and CYP2C19. M3 was produced by hydroxylation of M1, while M4 was produced by hydroxylation of M2; both hydroxylation reactions were catalyzed by CYP3A4. The formation of U1 was catalyzed by UGT2B7, but UGT1A4, UGT1A9, and UGT2B7 contributed to the formation of U2. In conclusion, DWP16001 is a substrate for CYP3A4, CYP2C19, UGT1A4, UGT1A9, and UGT2B7 enzymes. Overall, DWP16001 is weakly metabolized in human hepatocytes, but there is a potential for the pharmacokinetic modulation and drug–drug interactions, involved in the responsible metabolizing enzymes of DWP16001 in humans.

Dose-dependent glucosuria of DWP16001, a novel selective SGLT-2 inhibitor, in healthy subjects

“…The fraction of DWP16001 excreted in an unchanged form in the urine was less than 2.5%. This finding is consistent with nonclinical data that show that DWP16001 is extensively metabolized and mainly eliminated through a non-renal pathway in humans [17]. SGLT2 inhibitors have controversial effects on kidney injuries, but the PK characteristics of DWP16001 might be less affected by renal impairment.…”

“…In an in vitro study, DWP16001 selectively inhibited SGLT2 and showed high potency for SGLT2 inhibition-more than 1000 times that of SGLT1. DWP16001 is mainly metabolized by CYP3A4 and CYP2C19 into active metabolites M1 and M2 or via glucuronidation by UGT2B7, UGT1A4, and UGT1A9 [17]. The inhibitory potency of these metabolites for SGLT-2 was 10 times weaker than that of parent molecule.…”

Pharmacokinetics and Tissue Distribution of Enavogliflozin in Mice and Rats

“…UGT1A4, UGT1A9, and UGT2B7 participated in the formation of glucuronide conjugates (U1, U2). These latter metabolites were also found in the hepatocytes of all the species, but the intensity of the respective metabolites were different depending on the species [23]. In vitro hepatic clearance was calculated as 36.5 mL/min/kg for mouse, 8.3 mL/min/kg for rats, 17.9 mL/min/kg for dogs, 18.5 mL/min/kg for monkeys, and 4.5 mL/min/kg for human cells [23].…”

“…In a study for investigating drug metabolism of enavogliflozin in hepatocytes from mouse, rat, dog, monkey, and human, it showed species-different metabolism [23]. Kim et al identified five phase I metabolites from hepatocytes including, two monohydroxylated metabolites for which CYP3A4 and CYP2C19 were mainly involved, and three dihydroxylated metabolites.…”

“…Kim et al identified five phase I metabolites from hepatocytes including, two monohydroxylated metabolites for which CYP3A4 and CYP2C19 were mainly involved, and three dihydroxylated metabolites. Five glucuronide metabolites of enavogliflozin were identified, and UGT1A4 and UGT2B7 were primarily involved in the formation of these glucuronide metabolites [23]. Major phase I metabolites-M1 (6-hydroxy envogliflozin), M3 (subsequent oxidation of M1), and M2 (hydroxylation at the cyclopropyl benzene moiety)-were all found in mouse, rat, dog, monkey, and human hepatocytes.…”