[Show abstract][Hide abstract] ABSTRACT: 8-(N-2-hydroxy-5-chlorobenzoyl)-amino-caprylic acid (5-CNAC), a compound lacking pharmacological activity enhances the absorption of salmon calcitonin, when co-administered. Disposition and biotransformation of 5-CNAC was studied in six healthy postmenopausal women following a single oral dose of 200mg (14)C-radiolabeled 5-CNAC (as disodium monohydrate salt). Blood, plasma, urine and feces collected over 7 days were analyzed for radioactivity. Metabolite profiles were determined in plasma and excreta and metabolite structures were elucidated by LC-MS/MS, LC-(1)H NMR, enzymatic methods and by comparison with reference compounds. Oral 5-CNAC was safe and well tolerated in this study population. 5-CNAC absorption was rapid (t(max)=0.5h; C(max)=9.00 ± 2.74 μM (mean ± SD, n=6) and almost complete. The elimination half-life (t(½)) was 1.5 ± 1.1h. The radioactive dose was excreted mainly in urine (≥ 90%) in form of metabolites and 0.071% as intact 5-CNAC. Excretion of radioactivity in feces was minor and mostly as metabolites (<3%). Radioactivity in plasma reached C(max) (35.4 ± 7.9 μM) at 0.75 h and declined with a half-life of 13.9 ± 4.3h. 5-CNAC accounted for 5.8% of the plasma radioactivity AUC(0-24h). 5-CNAC was rapidly cleared from the systemic circulation, primarily by metabolism. Biotransformation of 5-CNAC involved: (a) stepwise degradation of the octanoic acid side chain and (b) conjugation of 5-CNAC and metabolites with glucuronic acid at the 2-phenolic hydroxyl group. The metabolism of 5-CNAC in vivo could be reproduced in vitro in human hepatocytes. No metabolism of 5-CNAC was observed in human liver microsomes.
European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2012; 47(1):44-55. DOI:10.1016/j.ejps.2012.04.023 · 3.35 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Aliskiren (2(S),4(S),5(S),7(S)-N-(2-carbamoyl-2-methylpropyl)-5-amino-4-hydroxy-2,7-diisopropyl-8-[4-methoxy-3-(3-methoxypropoxy)phenyl]-octanamid hemifumarate) is the first in a new class of orally active, nonpeptide direct renin inhibitors developed for the treatment of hypertension. The absorption, distribution, metabolism, and excretion of [(14)C]aliskiren were investigated in four healthy male subjects after administration of a single 300-mg oral dose in an aqueous solution. Plasma radioactivity and aliskiren concentration measurements and complete urine and feces collections were made for 168 h postdose. Peak plasma levels of aliskiren (C(max)) were achieved between 2 and 5 h postdose. Unchanged aliskiren represented the principal circulating species in plasma, accounting for 81% of total plasma radioactivity (AUC(0-infinity)), and indicating very low exposure to metabolites. Terminal half-lives for radioactivity and aliskiren in plasma were 49 h and 44 h, respectively. Dose recovery over 168 h was nearly complete (91.5% of dose); excretion occurred almost completely via the fecal route (90.9%), with only 0.6% recovered in the urine. Unabsorbed drug accounted for a large dose proportion recovered in feces in unchanged form. Based on results from this and from previous studies, the absorbed fraction of aliskiren can be estimated to approximately 5% of dose. The absorbed dose was partly eliminated unchanged via the hepatobiliary route. Oxidized metabolites in excreta accounted for at least 1.3% of the radioactive dose. The major metabolic pathways for aliskiren were O-demethylation at the phenyl-propoxy side chain or 3-methoxy-propoxy group, with further oxidation to the carboxylic acid derivative.
Drug Metabolism and Disposition 09/2007; 35(8):1418-28. DOI:10.1124/dmd.106.013797 · 3.25 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The highly positively charged, cell-penetrating beta3-octaarginine has been prepared with a radioactive label by acetylation at the N-terminus with a doubly (14)C-labeled acetyl group ((14)CH3-(14)CO). With the radioactive compound, an ADME study (Absorption, Distribution, Metabolism, Excretion) was performed in male rats following an intravenous or oral dose of 1 mg/kg. Sampling was carried out after periods ranging from 5 min to 4 d or 7 d for blood/excretia and quantitative whole-body autoradioluminography (QWBA), respectively. After p.o. dosing, no systemic exposure to peptide-related radioactivity was observed, and the dose was completely excreted in the feces within 24 h suggesting the absence of relevant absorption; less than 3% of the i.v. dose was excreted from the animals within 4 d. Blood levels, after i.v. dosing, dropped within 4 d to less than 2% of Cmax and decreased afterwards only very slowly. No metabolites were observed in the systemic circulation. QWBA Data indicated that the distribution of the acetyl-beta-octaarginine-related radioactivity in the organs and tissues shifted over time. Notably, after 7 d, the highest concentration was measured in the lymph nodes, and the largest amount was found in the liver. A comparison with the results of two previous ADME investigations of beta-peptides (cf. Table 1) reveals that the distribution of the compounds within the animals is structure-dependent, and that there is a full range from oral availability with rather rapid excretion (of a tetrapeptide) to essentially complete lack of both oral absorption and excretion after i.v. administration (of a highly charged octapeptide). A discussion is presented about the in vivo stability and 'drug-ability' of peptides. In general, beta-peptides bearing proteinogenic side chains are compared with peptides consisting entirely of D-alpha-amino acid residues (the enantiomers of the 'natural' building blocks), and suggestions are made regarding a possible focus of future biomedical investigations with beta-peptides.
[Show abstract][Hide abstract] ABSTRACT: The solid-phase synthesis and an ADME investigation with albino and pigmented male rats of the doubly 14C-labelled beta/alpha-tetrapeptide derivative Ac-beta3 hTyr-(D)Trp-beta3 hLys-beta3 hThr-lactone (3; Fig. 3) are described. After intravenous (i.v.) and peroral (p.o.) administration of the peptide, its concentration in blood and plasma, its tissue distribution, and the metabolism and the excretion of the peptide were analyzed over a period of up to 7 days post dose. The tetrapeptide in its ring opened form, 5, has a bioavailability of ca. 25%; radioactivity is distributed in the animals in an organ-specific way, and the compound appears to pass the blood-brain barrier to a very small extent, if at all (Tables 1-3 and Figs. 2-6). Excretion (37% renal, 44% fecal, including biliary) of the tetrapeptide 4 days after i.v. administration is almost complete, with only 4.3% remaining in the carcass; 4 days after p.o. administration 97% of the dose has been excreted in the feces. Radiochromatograms taken of plasma (0.5 and 24 h after i.v. dosing) and of urine and feces extracts (0-48 h collected) reveal the presence of lactone 3 and/or the corresponding hydroxy acid 5 with essentially no or very minor other peaks, respectively, representing possible metabolites (Tables 4-6, and Fig. 7 and 8). A comparison with a previous ADME investigation of a beta-nonapeptide show that--except for the lack of metabolism--all aspects of exposure, distribution, and elimination are different (structure-specific properties). The investigated tetrapeptide 3 is a potent and highly specific agonist of the somatostatin receptor hsst4, rendering the results described herein promising for diagnostic and therapeutic applications of beta-peptides.
[Show abstract][Hide abstract] ABSTRACT: In vitro studies: In CaCo-2 cell monolayers the β-nonapeptide H(β-HAla-β-HLys-β-HPhe)3-OH·4HCl (1), 14C-labeled on both C atoms of the CH2–CO moiety of the central β-HPhe residue, showed a low intrinsic permeability (<1%) and is subject to a prominent efflux system. The β-peptide (1) binds to human and rat plasma protein in vitro independent of the concentration of 1 and of the species (30–36% bound fraction at 50, 500, and 5000 ng/ml), and has only low affinity for the corresponding blood cells (less than 5% of compound 1 in blood cells).
In vivo studies: The in vivo pharmacokinetic characteristics after i.v. administration of 5 mg/kg (to male rats and to bile-duct-operated rats) were: (i) negligible in vivo biotransformation of 1 (in urine, plasma and feces unchanged 1 represented virtually the only compound-related molecule); (ii) rapid initial decline (0–8 h post dose) of levels of compound 1 in blood and plasma followed by a slower decline (8–96 h post dose); (iii) in non-operated animals after 96 h only 38% of the dose was excreted and after 168 h 49% of the dose was found remaining in the carcass; elimination through the intestine wall represented the major elimination pathway in non-operated animals while in bile-duct-cannulated animals biliary excretion was not found to contribute substantially to elimination (iv) quantitative whole-body autoradioluminography (QWBAL) investigations revealed that the kidney was by far the most important target organ of distribution; other tissues with high concentrations of compound-related radioactivity were cartilage, lymph nodes, and liver, whereas lowest levels were found in white fat and in the brain. After p.o. administration (10 mg/kg) negligible radioactivity was observed in the systemic circulation, indicating negligible absorption; essentially the entire oral dose was recovered unchanged in feces collected over a period of 96 h. Copyright
[Show abstract][Hide abstract] ABSTRACT: 5-CNAC (8-(N-2-hydroxy-5-chloro-benzoyl)-amino-caprylic acid) is used an excipient and as a delivery agent to enhance the oral absorption of salmon calcitonin (sCT). 5-CNAC exerts no pharmacological effects. 5-CNAC was developed by Emisphere's Eligen Technology and licensed in by Novartis. The present study was performed to investigate the absorption, pharmacokinetics, metabolism and excretion (ADME) of 14C‑radiolabeled 5-CNAC in six healthy post-menopausal female subjects, following a single oral dose of 200 mg [14C]5‑CNAC di-sodium monohydrate salt. The compound was administered in a gelatin capsule, without sCT. Subjects were dosed in a fasted state.
There were no serious adverse events, and none of the subjects discontinued the study prematurely. Absorption was rapid (tmax= 0.5 h; Cmax= 2.30.86 g/mL) and virtually complete. Unchanged 5‑CNAC contributed to 6% of the 14C‑AUC0‑24h in plasma; the rest of the 14C‑AUC0‑24h was due to metabolites. Elimination of 5-CNAC was rapid (t= 1.51.1 h) and occurred mainly by metabolism. 5‑CNAC appeared to undergo a substantial first pass metabolism, mainly by stepwise degradation of the octanoic acid side chain of 5‑CNAC, analogous to the fatty-acid -oxidation pathway. In plasma at 0.5 hours post-dose, apart from 5-CNAC, six metabolites were observed. Three of them (M5, M3, M2) were main metabolites: the 2‑O‑glucuronide of the butanoic acid metabolite of 5‑CNAC (M5) and of 5‑CNAC itself (M3), and the butanoic acid metabolite of 5-CNAC (M2). Overall, about 30%, 21%, and 8% of 14C‑AUC0‑24h were accounted for by the M5, M3, and M2, respectively. 5‑CNAC-related radioactivity disappeared from plasma with a terminal half-life of 144 hours. 5‑chlorosalicylic acid was observed as a minor metabolite which disappeared from plasma beyond 24 hours post-dose almost in parallel to total radioactivity. The biotransformation of 5-CNAC involves essentially two major metabolic pathways, namely: (i) stepwise degradation of the octanoic acid side chain of 5-CNAC, followed by 2‑O‑glucuronidation and (ii) direct conjugation of 5‑CNAC at the 2‑hydroxy group.
*= position of the carbon-14 radiolabel
Excretion of the radioactivity was almost complete (~ 90% of dose) after 1 day, and occurred predominantly via urine. Fecal excretion was minor (≤ 2%). In urine and feces, < 0.08% and ~ 0.3% of dose were excreted as unchanged 5-CNAC, respectively. By 7 days, the mass balance was close to complete (94.03.5%).
Overall, 5-CNAC is predominantly cleared by metabolism, primarily through ‑oxidation pathway (68% of dose), and additionally through glucuronidation at the 2-O-phenolic position of 5‑CNAC and its metabolites (79%).
10th European Regional International society for the study of xenobiotics Meeting;