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  • Hepatic, intestinal, renal, and plasma hydrolysis of prodrugs in human, cynomolgus monkey, dog, and rat: implications for in vitro-in vivo extrapolation of clearance of prodrugs.

Hepatic, intestinal, renal, and plasma hydrolysis of prodrugs in human, cynomolgus monkey, dog, and rat: implications for in vitro-in vivo extrapolation of clearance of prodrugs.

Drug metabolism and disposition: the biological fate of chemicals (2014-07-06)
Haruka Nishimuta, J Brian Houston, Aleksandra Galetin
ABSTRACT

Hydrolysis plays an important role in metabolic activation of prodrugs. In the current study, species and in vitro system differences in hepatic and extrahepatic hydrolysis were investigated for 11 prodrugs. Ten prodrugs in the data set are predominantly hydrolyzed by carboxylesterases (CES), whereas olmesartan medoxomil is also metabolized by carboxymethylenebutenolidase (CMBL) and paraoxonase. Metabolic stabilities were assessed in cryopreserved hepatocytes, liver S9 (LS9), intestinal S9 (IS9), kidney S9 (KS9), and plasma from human, monkey, dog, and rat. Of all the preclinical species investigated, monkey intrinsic hydrolysis clearance obtained in hepatocytes (CLint,hepatocytes) were the most comparable to human hepatocyte data. Perindopril and candesartan cilexetil showed the lowest and highest CLint,hepatocytes, respectively, regardless of the species investigated. Scaled intrinsic hydrolysis clearance obtained in LS9 were generally higher than CLint,hepatocytes in all species investigated, with the exception of dog. In the case of human and dog intestinal S9, hydrolysis intrinsic clearance could not be obtained for CES1 substrates, but hydrolysis for CES2 and CMBL substrates was detected in IS9 and KS9 from all species. Pronounced species differences were observed in plasma; hydrolysis of CES substrates was only evident in rat. Predictability of human hepatic intrinsic clearance (CLint,h) was assessed for eight CES1 substrates using hepatocytes and LS9; extrahepatic hydrolysis was not considered due to high stability of these prodrugs in intestinal and kidney S9. On average, predicted oral CLint,h from hepatocyte data represented 20% of the observed value; the underprediction was pronounced for high-clearance prodrugs, consistent with the predictability of cytochrome P450/conjugation clearance from this system. Prediction bias was less apparent with LS9, in particular for high-clearance prodrugs, highlighting the application of this in vitro system for investigation of prodrugs.

MATERIALS
Product Number
Brand
Product Description

Oseltamivir phosphate (impurity B free), European Pharmacopoeia (EP) Reference Standard
Benazepril for system suitability, European Pharmacopoeia (EP) Reference Standard
Quinapril hydrochloride, European Pharmacopoeia (EP) Reference Standard
Quinapril for system suitability, European Pharmacopoeia (EP) Reference Standard
Supelco
Mycophenolate Mofetil, Pharmaceutical Secondary Standard; Certified Reference Material
Candesartan cilexetil, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Candesartan cilexetil, ≥98% (HPLC)
Sigma-Aldrich
Mycophenolate mofetil, ≥98% (HPLC)
USP
Mycophenolate mofetil, United States Pharmacopeia (USP) Reference Standard
Mycophenolate mofetil, European Pharmacopoeia (EP) Reference Standard
Perindopril for peak identification, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Quinapril hydrochloride, ≥98% (HPLC), solid
Sigma-Aldrich
Ramipril, ≥98% (HPLC)
Sigma-Aldrich
Benazepril hydrochloride, ≥98% (HPLC), solid
Sigma-Aldrich
Perindopril erbumine
Benazepril hydrochloride, European Pharmacopoeia (EP) Reference Standard
Ramipril, European Pharmacopoeia (EP) Reference Standard
USP
Oseltamivir phosphate, United States Pharmacopeia (USP) Reference Standard
Quinapril for peak identification, European Pharmacopoeia (EP) Reference Standard
Candesartan cilexetil for peak identification, European Pharmacopoeia (EP) Reference Standard
Supelco
Ramipril, Pharmaceutical Secondary Standard; Certified Reference Material
Candesartan cilexetil for system suitability, European Pharmacopoeia (EP) Reference Standard
USP
Ramipril, United States Pharmacopeia (USP) Reference Standard
Perindopril tert-butylamine, European Pharmacopoeia (EP) Reference Standard
Perindopril for stereochemical purity, European Pharmacopoeia (EP) Reference Standard
Mycophenolate mofetil for peak identification, European Pharmacopoeia (EP) Reference Standard