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Rational design of agonists for bitter taste receptor TAS2R14: from modeling to bench and back

Abstract

Human bitter taste receptors (TAS2Rs) are a subfamily of 25 G protein-coupled receptors that mediate bitter taste perception. TAS2R14 is the most broadly tuned bitter taste receptor, recognizing a range of chemically diverse agonists with micromolar-range potency. The receptor is expressed in several extra-oral tissues and is suggested to have physiological roles related to innate immune responses, male fertility, and cancer. Higher potency ligands are needed to investigate TAS2R14 function and to modulate it for future clinical applications. Here, a structure-based modeling approach is described for the design of TAS2R14 agonists beginning from flufenamic acid, an approved non-steroidal anti-inflammatory analgesic that activates TAS2R14 at sub-micromolar concentrations. Structure-based molecular modeling was integrated with experimental data to design new TAS2R14 agonists. Subsequent chemical synthesis and in vitro profiling resulted in new TAS2R14 agonists with improved potency compared to the lead. The integrated approach provides a validated and refined structural model of ligand–TAS2R14 interactions and a general framework for structure-based discovery in the absence of closely related experimental structures.

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References

  1. 1.

    Jacobson KA (2015) New paradigms in GPCR drug discovery. Biochem Pharmacol 98(4):541–555

  2. 2.

    Hauser AS, Attwood MM, Rask-Andersen M, Schioth HB, Gloriam DE (2017) Trends in GPCR drug discovery: new agents, targets and indications. Nat Rev Drug Discov 16(12):829–842. https://doi.org/10.1038/nrd.2017.178

  3. 3.

    Pluskal T, Weng JK (2018) Natural product modulators of human sensations and mood: molecular mechanisms and therapeutic potential. Chem Soc Rev 47(5):1592–1637. https://doi.org/10.1039/c7cs00411g

  4. 4.

    Di Pizio A, Ben Shoshan-Galeczki Y, Hayes JE, Niv MY (2018) Bitter and sweet tasting molecules: it’s complicated. Neurosci Lett. https://doi.org/10.1016/j.neulet.2018.04.027

  5. 5.

    Wang M, Yao Y, Kuang D, Hampson DR (2006) Activation of family C G-protein-coupled receptors by the tripeptide glutathione. J Biol Chem 281(13):8864–8870. https://doi.org/10.1074/jbc.M512865200

  6. 6.

    Di Pizio A, Niv MY (2014) Computational studies of smell and taste receptors. Isr J Chem 54(8–9):1205–1218. https://doi.org/10.1002/ijch.201400027

  7. 7.

    Takeda S, Kadowaki S, Haga T, Takaesu H, Mitaku S (2002) Identification of G protein-coupled receptor genes from the human genome sequence. FEBS Lett 520(1):97–101. https://doi.org/10.1016/S0014-5793(02)02775-8

  8. 8.

    Behrens M, Meyerhof W (2010) Oral and extraoral bitter taste receptors. Results Probl Cell Differ 52:87–99

  9. 9.

    Trivedi BP (2012) Neuroscience: hardwired for taste. Nature 486(7403):S7–S9. https://doi.org/10.1038/486S7a

  10. 10.

    Clark AA, Dotson CD, Elson AET, Voigt A, Boehm U, Meyerhof W, Steinle NI, Munger SD (2015) TAS2R bitter taste receptors regulate thyroid function. FASEB J 29(1):164–172. https://doi.org/10.1096/fj.14-262246

  11. 11.

    Lee R, Cohen N (2015) Taste receptors in innate immunity. Cell Mol Life Sci 72(2):217–236. https://doi.org/10.1007/s00018-014-1736-7

  12. 12.

    Malki A, Fiedler J, Fricke K, Ballweg I, Pfaffl MW, Krautwurst D (2015) Class I odorant receptors, TAS1R and TAS2R taste receptors, are markers for subpopulations of circulating leukocytes. J Leukoc Biol. https://doi.org/10.1189/jlb.2a0714-331rr

  13. 13.

    Di Pizio A, Behrens M, Krautwurst D (2019) Beyond the flavour: the potential druggability of chemosensory G protein-coupled receptors. Int J Mol Sci. https://doi.org/10.3390/ijms20061402

  14. 14.

    Drewnowski A, Gomez-Carneros C (2000) Bitter taste, phytonutrients, and the consumer: a review. Am J Clin Nutr 72(6):1424–1435

  15. 15.

    Thawabteh A, Lelario F, Scrano L, Bufo SA, Nowak S, Behrens M, Di Pizio A, Niv MY, Karaman R (2018) Bitterless guaifenesin prodrugs—design, synthesis, characterization, in vitro kinetics and bitterness studies. Chem Biol Drug Des. https://doi.org/10.1111/cbdd.13409

  16. 16.

    Lee SJ, Depoortere I, Hatt H (2019) Therapeutic potential of ectopic olfactory and taste receptors. Nat Rev Drug Discov 18(2):116–138. https://doi.org/10.1038/s41573-018-0002-3

  17. 17.

    Stern L, Giese N, Hackert T, Strobel O, Schirmacher P, Felix K, Gaida MM (2018) Overcoming chemoresistance in pancreatic cancer cells: role of the bitter taste receptor T2R10. J Cancer 9(4):711–725. https://doi.org/10.7150/jca.21803

  18. 18.

    Hilger D, Masureel M, Kobilka BK (2018) Structure and dynamics of GPCR signaling complexes. Nat Struct Mol Biol 25(1):4–12. https://doi.org/10.1038/s41594-017-0011-7

  19. 19.

    Manglik A, Lin H, Aryal DK, McCorvy JD, Dengler D, Corder G, Levit A, Kling RC, Bernat V, Hübner H, Huang X-P, Sassano MF, Giguère PM, Löber S, Da D, Scherrer G, Kobilka BK, Gmeiner P, Roth BL, Shoichet BK (2016) Structure-based discovery of opioid analgesics with reduced side effects. Nature 537(7619):185–190. https://doi.org/10.1038/nature19112

  20. 20.

    Männel B, Jaiteh M, Zeifman A, Randakova A, Möller D, Hübner H, Gmeiner P, Carlsson J (2017) Structure-guided screening for functionally selective D2 dopamine receptor ligands from a virtual chemical library. ACS Chem Biol 12(10):2652–2661. https://doi.org/10.1021/acschembio.7b00493

  21. 21.

    Di Pizio A, Kruetzfeldt LM, Cheled-Shoval S, Meyerhof W, Behrens M, Niv MY (2017) Ligand binding modes from low resolution GPCR models and mutagenesis: chicken bitter taste receptor as a test-case. Sci Rep 7(1):8223. https://doi.org/10.1038/s41598-017-08344-9

  22. 22.

    Brockhoff A, Behrens M, Niv MY, Meyerhof W (2010) Structural requirements of bitter taste receptor activation. Proc Natl Acad Sci USA 107(24):11110–11115. https://doi.org/10.1073/pnas.0913862107

  23. 23.

    Born S, Levit A, Niv MY, Meyerhof W, Behrens M (2013) The human bitter taste receptor TAS2R10 is tailored to accommodate numerous diverse ligands. J Neurosci 33(1):201–213

  24. 24.

    Di Pizio A, Shy N, Behrens M, Meyerhof W, Niv MY (2018) Molecular features underlying selectivity in chicken bitter taste receptors. Front Mol Biosci 5:6. https://doi.org/10.3389/fmolb.2018.00006

  25. 25.

    Sandal M, Behrens M, Brockhoff A, Musiani F, Giorgetti A, Carloni P, Meyerhof W (2015) Evidence for a transient additional ligand binding site in the TAS2R46 bitter taste receptor. J Chem Theory Comput 11(9):4439–4449. https://doi.org/10.1021/acs.jctc.5b00472

  26. 26.

    Biarnés X, Marchiori A, Giorgetti A, Lanzara C, Gasparini P, Carloni P, Born S, Brockhoff A, Behrens M, Meyerhof W (2010) Insights into the binding of phenylthiocarbamide (PTC) agonist to its target human TAS2R38 bitter receptor. PLoS One 5(8):e12394. https://doi.org/10.1371/journal.pone.0012394

  27. 27.

    Jaggupilli A, Singh N, De Jesus VC, Gounni MS, Dhanaraj P, Chelikani P (2018) Chemosensory bitter taste receptors (T2Rs) are activated by multiple antibiotics. FASEB J. https://doi.org/10.1096/fj.201800521rr

  28. 28.

    Hariri BM, McMahon DB, Chen B, Freund JR, Mansfield CJ, Doghramji LJ, Adappa ND, Palmer JN, Kennedy DW, Reed DR, Jiang P, Lee RJ (2017) Flavones modulate respiratory epithelial innate immunity: anti-inflammatory effects and activation of the T2R14 receptor. J Biol Chem 292(20):8484–8497

  29. 29.

    Gentiluomo M, Crifasi L, Luddi A, Locci D, Barale R, Piomboni P, Campa D (2017) Taste receptor polymorphisms and male infertility. Hum Reprod 32(11):2324–2331. https://doi.org/10.1093/humrep/dex305

  30. 30.

    Levit A, Nowak S, Peters M, Wiener A, Meyerhof W, Behrens M, Niv MY (2014) The bitter pill: clinical drugs that activate the human bitter taste receptor TAS2R14. FASEB J 28(3):1181–1197. https://doi.org/10.1096/fj.13-242594

  31. 31.

    Behrens M, Gu M, Fan SJ, Huang C, Meyerhof W (2018) Bitter substances from plants used in traditional Chinese medicine exert biased activation of human bitter taste receptors. Chem Biol Drug Des 91(2):422–433. https://doi.org/10.1111/cbdd.13089

  32. 32.

    Di Pizio A, Niv MY (2015) Promiscuity and selectivity of bitter molecules and their receptors. Bioorg Med Chem 23(14):4082–4091. https://doi.org/10.1016/j.bmc.2015.04.025

  33. 33.

    Dagan-Wiener A, Di Pizio A, Nissim I, Bahia MS, Dubovski N, Margulis E, Niv MY (2018) BitterDB: taste ligands and receptors database in 2019. Nucleic Acids Res. https://doi.org/10.1093/nar/gky974

  34. 34.

    Karaman R, Nowak S, Di Pizio A, Kitaneh H, Abu-Jaish A, Meyerhof W, Niv MY, Behrens M (2016) Probing the binding pocket of the broadly tuned human bitter taste receptor TAS2R14 by chemical modification of cognate agonists. Chem Biol Drug Des 88(1):66–75. https://doi.org/10.1111/cbdd.12734

  35. 35.

    Ballesteros JA, Weinstein H (1995) [19] Integrated methods for the construction of three-dimensional models and computational probing of structure-function relations in G protein-coupled receptors. Methods Neurosci 25(25):366–428. https://doi.org/10.1016/s1043-9471(05)80049-7

  36. 36.

    Nowak S, Di Pizio A, Levit A, Niv MY, Meyerhof W, Behrens M (2018) Reengineering the ligand sensitivity of the broadly tuned human bitter taste receptor TAS2R14. Biochim Biophys Acta 1862(10):2162–2173. https://doi.org/10.1016/j.bbagen.2018.07.009

  37. 37.

    Lassalas P, Gay B, Lasfargeas C, James MJ, Tran V, Vijayendran KG, Brunden KR, Kozlowski MC, Thomas CJ, Smith AB 3rd, Huryn DM, Ballatore C (2016) Structure property relationships of carboxylic acid isosteres. J Med Chem 59(7):3183–3203

  38. 38.

    Fang Z, Yn Song, Zhan P, Zhang Q, Liu X (2014) Conformational restriction: an effective tactic in ‘follow-on’-based drug discovery. Future Med Chem 6(8):885–901. https://doi.org/10.4155/fmc.14.50

  39. 39.

    Ueda T, Ugawa S, Yamamura H, Imaizumi Y, Shimada S (2003) Functional interaction between T2R taste receptors and G-protein α subunits expressed in taste receptor cells. J Neurosci 23(19):7376–7380. https://doi.org/10.1523/JNEUROSCI.23-19-07376.2003

  40. 40.

    Behrens M, Blank K, Meyerhof W (2017) Blends of non-caloric sweeteners saccharin and cyclamate show reduced off-taste due to TAS2R bitter receptor inhibition. Cell Chem Biol 24(10):1199. https://doi.org/10.1016/j.chembiol.2017.08.004

  41. 41.

    Fish I, Stößel A, Eitel K, Valant C, Albold S, Hübner H, Möller D, Clark MJ, Sunahara RK, Christopoulos A, Shoichet BK, Gmeiner P (2017) Structure-based design and discovery of new M2 receptor agonists. J Med Chem 60(22):9239–9250. https://doi.org/10.1021/acs.jmedchem.7b01113

  42. 42.

    Liu H, Hofmann J, Fish I, Schaake B, Eitel K, Bartuschat A, Kaindl J, Rampp H, Banerjee A, Hübner H, Clark MJ, Vincent SG, Fisher JT, Heinrich MR, Hirata K, Liu X, Sunahara RK, Shoichet BK, Kobilka BK, Gmeiner P (2018) Structure-guided development of selective M3 muscarinic acetylcholine receptor antagonists. Proc Natal Acad Sci 115(47):12046–12050

  43. 43.

    Pegoli A, She X, Wifling D, Hübner H, Bernhardt G, Gmeiner P, Keller M (2017) Radiolabeled dibenzodiazepinone-type antagonists give evidence of dualsteric binding at the M2 muscarinic acetylcholine receptor. J Med Chem 60(8):3314–3334. https://doi.org/10.1021/acs.jmedchem.6b01892

  44. 44.

    Lachmann D, Studte C, Männel B, Hübner H, Gmeiner P, König B (2017) Photochromic dopamine receptor ligands based on dithienylethenes and fulgides. Chemistry 23(54):13423–13434. https://doi.org/10.1002/chem.201702147

  45. 45.

    Sommer T, Hübner H, El Kerdawy A, Gmeiner P, Pischetsrieder M, Clark T (2017) Identification of the beer component hordenine as food-derived dopamine D2 receptor agonist by virtual screening a 3D compound database. Sci Rep 7:44201. https://doi.org/10.1038/srep44201

  46. 46.

    Hübner H, Schellhorn T, Gienger M, Schaab C, Kaindl J, Leeb L, Clark T, Möller D, Gmeiner P (2016) Structure-guided development of heterodimer-selective GPCR ligands. Nat Commun 7:12298. https://doi.org/10.1038/ncomms12298

  47. 47.

    Conklin BR, Farfel Z, Lustig KD, Julius D, Bourne HR (1993) Substitution of three amino acids switches receptor specificity of Gq alpha to that of Gi alpha. Nature 363(6426):274–276. https://doi.org/10.1038/363274a0

  48. 48.

    Jiang LI, Collins J, Davis R, Lin KM, DeCamp D, Roach T, Hsueh R, Rebres RA, Ross EM, Taussig R, Fraser I, Sternweis PC (2007) Use of a cAMP BRET sensor to characterize a novel regulation of cAMP by the sphingosine 1-phosphate/G(13) pathway. J Biol Chem 282(14):10576–10584. https://doi.org/10.1074/jbc.M609695200

  49. 49.

    Du H, Brender JR, Zhang J, Zhang Y (2015) Protein structure prediction provides comparable performance to crystallographic structures in docking-based virtual screening. Methods 71:77–84. https://doi.org/10.1016/j.ymeth.2014.08.017

  50. 50.

    Congreve M, Oswald C, Marshall FH (2017) Applying structure-based drug design approaches to allosteric modulators of GPCRs. Trends Pharmacol Sci 38(9):837–847. https://doi.org/10.1016/j.tips.2017.05.010

  51. 51.

    Lyu J, Wang S, Balius TE, Singh I, Levit A, Moroz YS, O’Meara MJ, Che T, Algaa E, Tolmachova K, Tolmachev AA, Shoichet BK, Roth BL, Irwin JJ (2019) Ultra-large library docking for discovering new chemotypes. Nature 566(7743):224. https://doi.org/10.1038/s41586-019-0917-9

  52. 52.

    Roth BL, Irwin JJ, Shoichet BK (2017) Discovery of new GPCR ligands to illuminate new biology. Nat Chem Biol 13(11):1143–1151. https://doi.org/10.1038/nchembio.2490

  53. 53.

    Jaggupilli A, Howard R, Upadhyaya JD, Bhullar RP, Chelikani P (2016) Bitter taste receptors: novel insights into the biochemistry and pharmacology. Int J Biochem Cell B 77:184–196. https://doi.org/10.1016/j.biocel.2016.03.005

  54. 54.

    Venkatakrishnan AJ, Ma AK, Fonseca R, Latorraca NR, Kelly B, Betz RM, Asawa C, Kobilka BK, Dror RO (2019) Diverse GPCRs exhibit conserved water networks for stabilization and activation. Proc Natl Acad Sci USA 116(8):3288–3293. https://doi.org/10.1073/pnas.1809251116

  55. 55.

    Rajan KT, Hill AG, Barr A, Whitwell E (1967) Flufenamic acid in rheumatoid arthritis. Ann Rheum Dis 26(1):43

  56. 56.

    Ouellet M, Falgueyret J-P, Percival MD (2004) Detergents profoundly affect inhibitor potencies against both cyclo-oxygenase isoforms. Biochem J 377(3):675

  57. 57.

    Guinamard R, Simard C, Del Negro C (2013) Flufenamic acid as an ion channel modulator. Pharmacol Ther 138(2):272–284. https://doi.org/10.1016/j.pharmthera.2013.01.012

  58. 58.

    Féau C, Arnold LA, Kosinski A, Zhu F, Connelly M, Guy RK (2009) Novel flufenamic acid analogues as inhibitors of androgen receptor mediated transcription. ACS Chem Biol 4(10):834–843. https://doi.org/10.1021/cb900143a

  59. 59.

    Zhang X, Stevens RC, Xu F (2015) The importance of ligands for G protein-coupled receptor stability. Trends Biochem Sci 40(2):79–87. https://doi.org/10.1016/j.tibs.2014.12.005

  60. 60.

    Adeniji AO, Twenter BM, Byrns MC, Jin Y, Chen M, Winkler JD, Penning TM (2012) Development of potent and selective inhibitors of aldo-keto reductase 1C3 (type 5 17beta-hydroxysteroid dehydrogenase) based on N-phenyl-aminobenzoates and their structure-activity relationships. J Med Chem 55(5):2311–2323. https://doi.org/10.1021/jm201547v

  61. 61.

    Behrens M, Brockhoff A, Kuhn C, Bufe B, Winnig M, Meyerhof W (2004) The human taste receptor hTAS2R14 responds to a variety of different bitter compounds. Biochem Biophys Res Commun 319(2):479–485. https://doi.org/10.1016/j.bbrc.2004.05.019

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Acknowledgements

This research was supported in part by the German Research Foundation Grants Gm 13/12 (to P.G. and M.Y.N.), GRK 1910 (to P.G.), and ISF 494/16 (to M.Y.N). Lady Davis Fellowship and COST-STSM-CM1207 (GLISTEN) to A.D.P. are gratefully acknowledged. M.Y.N and A.D.P. participate in Mu.Ta.Lig—COST ACTION CA15135. The authors would like to thank Catherine Delaporte for the excellent technical assistance and Tamir Dingjan for the critical reading of the manuscript.

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Correspondence to Maik Behrens or Peter Gmeiner or Masha Y. Niv.

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Di Pizio, A., Waterloo, L.A.W., Brox, R. et al. Rational design of agonists for bitter taste receptor TAS2R14: from modeling to bench and back. Cell. Mol. Life Sci. 77, 531–542 (2020). https://doi.org/10.1007/s00018-019-03194-2

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Keywords

  • Bitter taste receptor
  • GPCRs
  • Drug design
  • Structure-based modeling
  • Bioisosteric replacement