Ex vivo gut-hepato-biliary organ perfusion model to characterize oral absorption, gut-wall metabolism, pre-systemic hepatic metabolism and biliary excretion; application to midazolam.

To date, characterization of the first-pass effect of orally administered drugs consisting of local intestinal absorption and metabolism, portal vein transport and hepatobiliary processes remains challenging. Aim of this study was to explore the applicability of a porcine ex-vivo perfusion model to study oral absorption, gut-hepatobiliary metabolism and biliary excretion of midazolam. Slaughterhouse procured porcine en bloc organs (n = 4), were perfused via the aorta and portal vein. After 120 min of perfusion, midazolam, atenolol, antipyrine and FD4 were dosed via the duodenum and samples were taken from the systemic- and portal vein perfusate, intestinal faecal effluent and bile to determine drug and metabolite concentrations. Stable arterial and portal vein flow was obtained and viability of the perfused organs was confirmed. After intraduodenal administration, midazolam was rapidly detected in the portal vein together with 1-OH midazolam (EG-pv of 0.16±0.1) resulting from gut wall metabolism through oxidation. In the intestinal faecal effluent, 1-OH midazolam and 1-OH midazolam glucuronide (EG-intestine 0.051±0.03) was observed resulting from local gut glucuronidation. Biliary elimination of midazolam (0.04±0.01 %) and its glucuronide (0.01±0.01 %) only minimally contributed to the enterohepatic circulation. More extensive hepatic metabolism (FH 0.35±0.07) over intestinal metabolism (FG 0.78±0.11) was shown, resulting in oral bioavailability of 0.27±0.05. Ex vivo perfusion demonstrated to be a novel approach to characterize pre-systemic extraction of midazolam by measuring intestinal as well as hepatic extraction. The model can generate valuable insights into the absorption and metabolism of new drugs.

Glucose and insulin metabolism in ruminating and veal calves fed high and low fat diets.

Holstein male calves were maintained on conventional (milk to 6 wk of age, fed grain and hay after weaning) and veal (milk replacer only) diets to 16 wk of age. Within each of these 2 physiological states (ruminating or non-ruminating), calves were fed low or high fat diets (ruminating: 3 and 10%; veal: 10 and 18%). Glucose tolerance tests were undertaken at 8 and 16 wk of age in each group. Basal concentrations (4 hr postfeeding) and areas under the response curves for plasma glucose and insulin were higher in veal calves (P < .0001). Ruminating calves fed higher fat utilized glucose more readily (smaller areas under the curves for both glucose and insulin, P < .10) than those fed lower fat. Age did not influence basal glucose concentrations (P > .10), but older calves had higher basal insulin (P < .0001) and greater areas under the curves (P < .0005) for both glucose and insulin after a glucose challenge. Rate of clearance (k) was greater in ruminating calves (P < .001). Though rate of clearance in veal calves was slower, larger plasma pool size caused veal calves on average to utilize glucose at a 15% greater rate per kg body weight than ruminating calves. Whereas fat concentration in the diets did not influence glucose metabolism in veal calves, the high lactose content (> 50% of diet dry matter) of veal diets induced severe insulin resistance in these calves.