[en] Free fatty acid-2 (FFA2) receptor is a G-protein coupled receptor of interest in the development of therapeutics in metabolic and inflammatory disease areas. The discovery and optimization of an N-thiazolylamide carboxylic acid FFA2 agonist scaffold is described. Dual key objectives were to i) evaluate the potential of this scaffold for lead optimization in particular with respect to safety de-risking physicochemical properties, i.e. lipophilicity and aromatic content, and ii) to demonstrate the utility of selected lead analogues from this scaffold in a pertinent in vivo model such as oral glucose tolerance test (OGTT). As such, a concomitant improvement in bioactivity together with lipophilic ligand efficiency (LLE) and fraction sp3 content (Fsp3) parameters guided these efforts. Compound 10 was advanced into studies in mice on the basis of its optimized profile vs initial lead 1 (ΔLLE = 0.3, ΔFsp3 = 0.24). Although active in OGTT, 10 also displayed similar activity in the FFA2-knockout mice. Given this off-target OGTT effect, we discontinued development of this FFA2 agonist scaffold.
Disciplines :
Chemistry
Author, co-author :
Hoveyda, Hamid R; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium. Electronic address: hamidhoveyda@hotmail.com
Fraser, Graeme L; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Zoute, Ludivine; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Dutheuil, Guillaume; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Schils, Didier ; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Brantis, Cyrille; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Lapin, Alexey; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Parcq, Julien; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Guitard, Sandra; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Lenoir, François; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Bousmaqui, Mohamed El; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Rorive, Sarah; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Hospied, Sandrine; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Blanc, Sébastien; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Bernard, Jérôme; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
Ooms, Frédéric ; Université de Liège - ULiège > HEC Liège Research > HEC Liège Research: Strategy & Performance for the Society ; Ogeda SA (formerly Euroscreen), 47, rue Adrienne Bolland, 6041 Gosselies, Belgium
McNelis, Joanne C; University of California, Department of Medicine, San Diego, CA 92161, USA
Olefsky, Jerrold M; University of California, Department of Medicine, San Diego, CA 92161, USA
This work was supported in part by the Ministry of Sustainable Development and Public Works, Walloon Region, Belgium (at Ogeda) and by Digestive and Kidney Diseases grants (DK033651, DK074868, DK063491 and DK09062) to J. M. O.
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It is worth to note in passing that the presence of N-thiazolylamide in both the foregoing agonist scaffold as well as in the Amgen allosteric agonist 3 (Fig. 1) was fortuitous in that we were unaware of the existence of 3 until well after our discovery and first round lead optimization of the N-thiazolylamide carboxylic acid scaffold discussed herein.
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Hudson, B.D., Due-Hansen, M.E., Christiansen, E., et al. Defining the molecular basis for the first potent and selective orthosteric agonists of the FFA2 free fatty acid receptor. J Biol Chem 288 (2013), 17296–17312 In contrast to other researchers who have used 8 extensively to explore various aspects of the FFA2 receptor pharmacology; e.g.
To be noted, Brown et al have recently reported that the initial lead 1 from the foregoing scaffold as well as Amgen's allosteric agonist 3 are active in ex vivo lipolysis assays in murine adipocyte tissue explants from wild-type mice (at 75 µM, 1% BSA), but not from FFA2 knock-out mice, as evidence that the ex-vivo lipolysis effect is FFA2 mediated; see: Brown AJ, Tsoulou C, Ward E, Gower E, Bhudia N, Chowdhury F, Dean TW, Faucher N, Gangar A, Dowell SJ, Pharmacological properties of acid N-thiazolylamide FFA2 agonists. Pharma Res. Per. 2015, e00141, 1-15. However, in repeat testing, we have found that in mice 1 is inactive in both in vivo lipolysis and in OGTT assays despite its acceptable PK profile (Cmax = 300 μM, 30 mg/kg oral dose). In addition, Hudson et al. (ref. 31) have reported that the early generation lead 8 exhibits inhibitory effect in ex vivo lipolysis assays using human and murine cell lines. Here again the effect was not demonstrated in vivo and nor was the mode-of-action verified through knock-out mice.
Compound 13 is the best lead structure herein due to its superior in vivo oral exposure (Table 2) in step with its improved physicochemical properties (Table 1) when compared to 10. Analogue 10 was used for the mechanism-of-action studies as this lead was discovered earlier and was deemed adequate for these studies.
Due-Hansen, M.E., Pandey, S.K., Christiansen, E., Andersen, R., Hansen, S.V.F., Ulven, T., A protocol for amide bond formation with electron deficient amines and sterically hindered substrates. Org Biomol Chem 14 (2016), 430–433 Due-Hansen et al advocate in situ acyl fluoride activation at 80 °C for more efficient coupling between N-cyclopropylthiazolamine (24-25, Scheme 1) and carboxylic acid 17 (racemate used in their work) versus HATU/DIEA amide coupling at ambient temperature. We obtain comparable yields with HATU/DIEA coupling at 60 °C, as also noted in our initial patent (see ref 3 above).
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