title: Assessment of enzymatic prodrug stability in human, dog and simulated intestinal fluids.
abstract: The aim of this study was to determine the stability of three ester prodrugs, chloramphenicol succinate, enalapril and candesartan cilexetil, in human proximal small intestinal fluid (HIF), dog proximal small intestinal fluids (DIF) and simulated intestinal fluid (FaSSIF), with the addition of pancreatin. The total protein content in the proximal jejunal fluids was determined in HIF and DIF, respectively. Candesartan cilexetil was significantly degraded in HIF (initial t(1/2(0-5 min))=5.4 ± 0.5 min) and in DIF (initial t(1/2(0-5 min))=5.7 ± 0.1 min), while chloramphenicol succinate and enalapril were stable in both media. The degradation of candesartan cilexetil was shown to be mediated by enzymes following Michaelis-Menten enzyme kinetics and was inhibited by addition of esterase inhibitors. The enzymatic capacity reflected by V(max) was 4-fold higher in DIF than in HIF and correlated to its 2-fold higher protein concentration. The degradation of candesartan cilexetil in the FaSSIF-pancreatin solution was slower (t(1/2)=207 ± 34 min) than the degradation in both HIF and DIF. Changing the pH to the enzyme optima or increasing the amount of pancreatin, increased the degradation rate of candesartan cilexetil, but not in the magnitude as in HIF. As a result, two in vitro models, based on in vivo intestinal fluids, were developed using candesartan cilexetil as a model drug. The DIF seems to be a reasonably good model for HIF, although the degradation capacity seems to be somewhat higher, possibly due to the higher enzyme concentration in DIF. Future investigations will develop novel enzymatic based in vitro models for rapid assessment and biopharmaceutical screening tools for prodrugs.
authors.affiliation: AstraZeneca R&D, Mölndal, Sweden.
authors.forename: A S
authors.initials: AS
authors.lastname: Borde
authors.forename: E M
authors.initials: EM
authors.lastname: Karlsson
authors.forename: K
authors.initials: K
authors.lastname: Andersson
authors.forename: K
authors.initials: K
authors.lastname: Björhall
authors.forename: H
authors.initials: H
authors.lastname: Lennernäs
authors.forename: B
authors.initials: B
authors.lastname: Abrahamsson
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D001562:Benzimidazoles / Q000032:analysis / Q000737:chemistry; D001702:Biopharmaceutics / Q000379:methods; D001713:Biphenyl Compounds / Q000032:analysis / Q000737:chemistry; D001826:Body Fluids / Q000737:chemistry*; D002701:Chloramphenicol / Q000031:analogs & derivatives / Q000032:analysis / Q000737:chemistry; D004285:Dogs; D004355:Drug Stability; D004656:Enalapril / Q000032:analysis / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007419:Intestinal Secretions / Q000737:chemistry* / Q000378:metabolism; D007421:Intestine, Small / Q000737:chemistry*; D008297:Male; D010194:Pancreatin / Q000737:chemistry; D011355:Prodrugs / Q000032:analysis* / Q000737:chemistry*; D013777:Tetrazoles / Q000032:analysis / Q000737:chemistry; D055815:Young Adult
chemical_list: D001562:Benzimidazoles; D001713:Biphenyl Compounds; D004791:Enzyme Inhibitors; D011355:Prodrugs; D013777:Tetrazoles; D002701:Chloramphenicol; D004656:Enalapril; D010194:Pancreatin; C077793:candesartan cilexetil; C026016:chloramphenicol succinate