High metabolic in vivo stability and bioavailability of a palmitoylated ghrelin receptor ligand assessed by mass spectrometry
| Autor: | Sylvia Els-Heindl, Martin von Bergen, Sven Baumann, Katja B. Kostelnik, Annette G. Beck-Sickinger, Nora Klöting |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Male
Stable isotopic labeling Lipoylation Clinical Biochemistry Drug Evaluation Preclinical Palmitic Acid Pharmaceutical Science Biological Availability Peptide Ligands Biochemistry Mass Spectrometry Isotopic labeling Structure-Activity Relationship Blood serum Drug Stability In vivo Drug Discovery Chlorocebus aethiops Inverse agonist Animals Humans Receptor Receptors Ghrelin Molecular Biology chemistry.chemical_classification Carbon Isotopes Mass spectrometry Chemistry Ghrelin receptor Organic Chemistry Selected reaction monitoring Rats Inbred Lew COS Cells Molecular Medicine Ghrelin Metabolic stability Peptides |
| Zdroj: | Kostelnik, K B, Els-Heindl, S, Kloting, N, Baumann, S, von Bergen, M & Beck-Sickinger, A G 2015, ' High metabolic in vivo stability and bioavailability of a palmitoylated ghrelin receptor ligand assessed by mass spectrometry ', Bioorganic & Medicinal Chemistry, vol. 23, no. 14, pp. 3925–3932 . https://doi.org/10.1016/j.bmc.2014.12.008 |
| Popis: | The constitutive activity of the ghrelin receptor is of high physiological and pathophysiological relevance. In-depth structure-activity relationship studies revealed a palmitoylated ghrelin receptor ligand that displays an in vitro binding affinity in the low nanomolar range. Activity studies revealed inverse agonistic as well as antagonistic properties and in vitro metabolic analysis indicated a high stability in blood serum and liver homogenate. For metabolic testing in vivo, a combined approach of stable isotopic labeling and mass spectrometry-based analysis was established. Therefore, a heavy isotopic version of the peptide containing a (13)C-labeled palmitic acid was synthesized and a 1:1 ratio of a (12)C/(13)C-peptide mixture was injected into rats. Biological samples were analyzed by multiple reaction monitoring allowing simultaneous peptide detection and quantification. Measurements revealed a suitable bioavailability over 24h in rat serum and subsequent high-resolution mass spectrometry investigations showed only negligible degradation and slow body clearance. Hence, this method combination allowed the identification and evaluation of a highly potent and metabolically stable ghrelin receptor ligand in vivo. |
| Databáze: | OpenAIRE |
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