Structure–Activity Relationships and Characterization of Highly Selective, Long-Acting, Peptide-Based Cholecystokinin 1 Receptor Agonists

Abstract

A group of peptide-based, long-acting, stable, highly selective cholecystokinin 1 receptor (CCK-1R) agonists with the potential to treat obesity has been identified and characterized, based on systematic investigation of synthetic CCK-8 analogues with N-terminal linkage to fatty acids. Sulfated Tyr in such compounds was stable in neutral buffer. CCK-1R selectivity was achieved mostly by introducing d-N-methyl-Asp instead of Asp at the penultimate position of CCK-8. Our compound 9 (NN9056) showed similar in vitro CCK-1R potency and CCK-1R affinity as CCK-8, very high selectivity for CCK-1R over the cholecystokinin 2 receptor (CCK-2R), strong reduction of food intake in lean pigs for up to 48 h after one subcutaneous injection without adverse effects, a plasma half-life of 113 h in minipigs after intravenous injection, and acceptable chemical stability in a neutral liquid formulation. In addition, we found a highly selective CCK-2R agonist by replacing Gly in a CCK-8 derivative with Glu.

Long-acting CCK analogue NN9056 lowers food intake and body weight in obese Göttingen Minipigs

“…In vitro assays CCK receptor binding and activation studies CCK analogue NN9056 and sulfated native human CCK-8 was synthesised at Novo Nordisk A/S [23]. All experiments were carried out essentially as previously described [25].…”

“…Based on the available data, it was hypothesised that treatment with a long-acting CCK-1 receptor-selective analogue with 24-h plasma exposure coverage would result in a significant and clinically relevant reduction in food intake and body weight (BW) without causing pancreatitis in humans. The acylated CCK analogue NN9056 is a highly CCK-1 receptor-selective compound with a terminal plasma half-life of approximately 110 h in young lean minipigs [23]. Due to the high expression of CCK-1 receptors in the pancreas, rodents and dogs were deselected as preclinical efficacy models, leaving pigs and non-human primates as relevant and well-characterised obesity models.…”

Structures of the human cholecystokinin receptors in complex with agonists and antagonists

“…The copyright holder for this preprint this version posted May 21, 2021. ; https://doi.org/10.1101/2021.05. 19…”

“…The copyright holder for this preprint this version posted May 21, 2021. ; https://doi.org/10.1101/2021.05. 19.444887 doi: bioRxiv preprint Gs. The CCKBR-Gq and CCKBR-Gi2 complexes were assembled in the presence of the CCKBR selective endogenous peptide gastrin-17 and their structures were determined by single-particle cryo-EM analysis with the overall resolution of 3.1 Å and 3.3 Å (Fig.…”

“…Highly selective non-peptidic antagonists of CCKAR such as devazepide [3S(-)-N(2,3-Dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4benzodiazepine-3-yl)-1H-indole-2-carboxamide] and lintitript [1][2] thiazole-2-yl)aminocarbonyl]indolyl) acetic acid] (Extended Data Fig. 1) were developed to treat gastrointestinal disorders, neuropathic pain and pancreatic cancer 17,18 , while highly selective CCKAR agonists such as NN9056, a modified CCK-8 with high selectivity and longacting properties, have been proposed as potential treatment for obesity 19,20 . Meanwhile, antagonists such as Z-360 or JB95008 and agonists such as ceruletide targeting CCKBR have been implicated in treating diabetes, anxiety and thyroid cancer, etc 21,22 .…”