title: Synthesis and evaluation of novel lipidated neuromedin U analogs with increased stability and effects on food intake.
abstract: Neuromedin U (NMU) is a 25 amino acid peptide expressed and secreted in the brain and gastrointestinal tract. Data have shown that peripheral administration of human NMU decreases food intake and body weight and improves glucose tolerance in mice, suggesting that NMU receptors constitute a possible anti-diabetic and anti-obesity drug target. However, the clinical use of native NMU is hampered by a poor pharmacokinetic profile. In the current study, we report in vitro and in vivo data from a series of novel lipidated NMU analogs. In vitro plasma stability studies of native NMU were performed to investigate the proteolytic stability and cleavage sites using LC-MS. Native NMU was found to be rapidly cleaved at the C-terminus between Arg(24) and Asn(25) , followed by cleavage between Arg(16) and Gly(17) . Lipidated NMU analogs were generated using solid-phase peptide synthesis, and in vitro potency was investigated using a human embryonic kidney 293-based inositol phosphate accumulation assay. All lipidated analogs had preserved in vitro activity on both NMU receptors with potency improving as the lipidation site was moved away from the receptor-interacting C-terminal octapeptide segment. In vivo efficacy was assessed in lean mice as reduction in food intake after acute subcutaneous administration of 1, 0.3, 0.1, and 0.03 µmol/kg. These lipidated NMU analogs prolonged the anorectic effect of NMU in a dose-dependent manner. This was likely an effect of improved pharmacokinetic properties because of improved vitro plasma stability. Accordingly, the data demonstrate that lipidated NMU analogs may represent drug candidates for the treatment of obesity.
authors.affiliation: Gubra ApS, Agern Alle 1, 2970, Hørsholm, Denmark; The Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Blegdamsvej 3B, 2200, Copenhagen N, Denmark.
authors.forename: Louise S
authors.initials: LS
authors.lastname: Dalbøge
authors.forename: Søren L
authors.initials: SL
authors.lastname: Pedersen
authors.forename: Søren B
authors.initials: SB
authors.lastname: van Witteloostuijn
authors.forename: Jakob E
authors.initials: JE
authors.lastname: Rasmussen
authors.forename: Kristoffer T G
authors.initials: KT
authors.lastname: Rigbolt
authors.forename: Knud J
authors.initials: KJ
authors.lastname: Jensen
authors.forename: Birgitte
authors.initials: B
authors.lastname: Holst
authors.forename: Niels
authors.initials: N
authors.lastname: Vrang
authors.forename: Jacob
authors.initials: J
authors.lastname: Jelsing
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004355:Drug Stability; D004435:Eating / Q000187:drug effects*; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D009479:Neuropeptides / Q000097:blood / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D055550:Protein Stability
chemical_list: D009479:Neuropeptides; C056900:neuromedin U
keywords: NMU; NMU analogs; acylation; food intake; lipidation; obesity