High metabolic in vivo stability and bioavailability of a palmitoylated ghrelin receptor ligand assessed by mass spectrometry

Kostelnik, Katja B.; Els‐Heindl, Sylvia; Klöting, Nora; Baumann, Sven; Bergen, Martin von; Beck‐Sickinger, Annette G.

doi:10.1016/j.bmc.2014.12.008

Cited by 18 publications

(27 citation statements)

References 48 publications

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“…Although the metabolic stability was not tested in the current study, it has previously been shown that adding polyethylene glycol to a peptide can increase metabolic stability. 89-90 By incorporating the polyethylene glycol-like PEDG20 into our design, it may increase the likelihood of identifying a suitable in vivo probe.…”

Section: Resultsmentioning

confidence: 99%

An in Vitro and in Vivo Investigation of Bivalent Ligands That Display Preferential Binding and Functional Activity for Different Melanocortin Receptor Homodimers

et al. 2016

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Pharmacological probes for the melanocortin receptors have been utilized for studying various disease states including cancer, sexual function disorders, Alzheimer's disease, social disorders, cachexia, and obesity. This study focused on the design and synthesis of bivalent ligands to target melanocortin receptor homodimers. Lead ligands increased binding affinity by 14- to 25-fold and increased cAMP signaling potency by 3- to 5-fold compared to their monovalent counterparts. Unexpectedly, different bivalent ligands showed preferences for particular melanocortin receptor subtypes depending on the linker that connected the binding scaffolds suggesting structural differences between the various dimer subtypes. Homobivalent compound 12 (CJL-1-140) possessed a functional profile that was unique from its monovalent counterparts providing evidence of the discrete effects of bivalent ligands. Lead compound 7 (CJL-1-87) significantly decreased feeding in mice after intracerebroventricular administration. To the best of our knowledge, this is the first report of a melanocortin bivalent ligand's in vivo physiological effects.

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Section: Resultsmentioning

confidence: 99%

An in Vitro and in Vivo Investigation of Bivalent Ligands That Display Preferential Binding and Functional Activity for Different Melanocortin Receptor Homodimers

et al. 2016

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“…Inositol phosphate (IP) accumulation assay for the ghrelin receptor was carried out as described previously . Shortly, COS‐7 cells stably transfected with the human GhrR were seeded out in 48‐well plates (80.000 cells/well) and cultured overnight.…”

Section: Methodsmentioning

confidence: 99%

A stable meta‐carborane enables the generation of boron‐rich peptide agonists targeting the ghrelin receptor

Worm

Els‐Heindl

Kellert

et al. 2018

Journal of Peptide Science

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Boron neutron capture therapy (BNCT) is a binary cancer therapy, which combines the biochemical targeting of a boron-containing drug with the regional localization of radiation treatment. Although the concept of BNCT has been known for decades, the selective delivery of boron into tumor cells remains challenging. G protein-coupled receptors that are overexpressed on cancer cells in combination with peptidic ligands can be potentially used as shuttle system for a tumor-directed boron uptake. In this study, we present the generation of short, boron-rich peptide conjugates that target the ghrelin receptor. Expression of the ghrelin receptor on various cancer cells makes it a viable target for BNCT. We designed a novel hexapeptide super-agonist that was modified with different specifically synthesized carborane monoclusters and tested for ghrelin receptor activation. A meta-carborane building block with a mercaptoacetic acid linker was found to be optimal for peptide modification, owing to its chemical stability and a suitable activation efficacy of the conjugate. The versatility of this carborane for the development of peptidic boron delivery agents was further demonstrated by the generation of highly potent, boron-loaded conjugates using the backbone of the known ghrelin receptor ligands growth hormone releasing peptide 6 and Ipamorelin.

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“…Neuropeptide Y (NPY), ghrelin and the inverse agonist KbFwLK(Pam)-NH 2 (b = β-(3-benzothienyl)-D-alanine) were synthesized by Fmoc/ tert -butyl strategy as described before2834. Fluorescent tracer [Dpr 3 -Oct; Dpr 16 -atto520]-ghrelin (Dpr, diaminopropionic acid; Oct, octanoic acid) for in vitro functionality assay was prepared similarly as described for a fluorescein-conjugated ghrelin analogue34.…”

Section: Methodsmentioning

confidence: 99%

“…Here, we report results on the molecular dynamics of the human growth hormone secretagogue (GHS) receptor, which plays a key role in the regulation of appetite and food intake and, therefore, represents an interesting target for intervening with eating disorders25. The ligand of the GHS receptor is the octanoylated growth hormone ghrelin consisting of 28 amino acids262728. We used solid-state NMR spectroscopy to study the molecular dynamics of the GHS receptor reconstituted into DMPC or POPC membranes.…”

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confidence: 99%

Expression, Functional Characterization, and Solid-State NMR Investigation of the G Protein-Coupled GHS Receptor in Bilayer Membranes

Schrottke

Kaiser

Vortmeier

et al. 2017

Sci Rep

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The expression, functional reconstitution and first NMR characterization of the human growth hormone secretagogue (GHS) receptor reconstituted into either DMPC or POPC membranes is described. The receptor was expressed in E. coli. refolded, and reconstituted into bilayer membranes. The molecule was characterized by 15N and 13C solid-state NMR spectroscopy in the absence and in the presence of its natural agonist ghrelin or an inverse agonist. Static 15N NMR spectra of the uniformly labeled receptor are indicative of axially symmetric rotational diffusion of the G protein-coupled receptor in the membrane. In addition, about 25% of the 15N sites undergo large amplitude motions giving rise to very narrow spectral components. For an initial quantitative assessment of the receptor mobility, 1H-13C dipolar coupling values, which are scaled by molecular motions, were determined quantitatively. From these values, average order parameters, reporting the motional amplitudes of the individual receptor segments can be derived. Average backbone order parameters were determined with values between 0.56 and 0.69, corresponding to average motional amplitudes of 40–50° of these segments. Differences between the receptor dynamics in DMPC or POPC membranes were within experimental error. Furthermore, agonist or inverse agonist binding only insignificantly influenced the average molecular dynamics of the receptor.

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High metabolic in vivo stability and bioavailability of a palmitoylated ghrelin receptor ligand assessed by mass spectrometry

Cited by 18 publications

References 48 publications

An in Vitro and in Vivo Investigation of Bivalent Ligands That Display Preferential Binding and Functional Activity for Different Melanocortin Receptor Homodimers

An in Vitro and in Vivo Investigation of Bivalent Ligands That Display Preferential Binding and Functional Activity for Different Melanocortin Receptor Homodimers

A stable meta‐carborane enables the generation of boron‐rich peptide agonists targeting the ghrelin receptor

Expression, Functional Characterization, and Solid-State NMR Investigation of the G Protein-Coupled GHS Receptor in Bilayer Membranes

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