Skip to main content
SpringerLink
Log in

UFP-101, a high affinity antagonist for the nociceptin/orphanin FQ receptor: radioligand and GTPγ35S binding studies

  • Original Article
  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Abstract

Studies of the pharmacology of nociceptin/orphanin FQ (N/OFQ) and its receptor (NOP) have been hampered by the lack of a range of high potency antagonists. In this study we have examined the effects of a novel N/OFQ analogue [Nphe1,Arg14,Lys15]N/OFQ NH2 hereafter referred to as UFP-101. [3H]N/OFQ competition binding and GTPγ35S binding assays were performed using CHO cells expressing the human NOP receptor (CHOhNOP). UFP-101 (pKi of 10.14±0.09) and a range of NOP selective agonists displaced [3H]N/OFQ binding with the following rank order of affinity: [Arg14,Lys15]N/OFQ>[(pF)Phe4]N/OFQ(1–13)NH2>N/OFQ(1–13)NH2>UFP-101>N/OFQ>Ro64–6198>[Nphe1]N/OFQ(1–13)NH2. N/OFQ, N/OFQ(1–13)NH2, [(pF)Phe4]N/OFQ(1-13)NH2, [Arg14,Lys15]N/OFQ and Ro64–6198 also produced a concentration dependent (pEC50 values of 8.75±0.11, 9.28±0.15, 9.69±0.04, 9.12±0.11 and 8.09±0.07 respectively) and saturable stimulation of GTPγ35S binding and all were full agonists. UFP-101 did not stimulate GTPγ35S binding per se, but produced a concentration dependent and parallel rightward shift in the concentration response curves to all agonists. UFP-101 yielded pA2 values in the range 8.4–9.0. For comparison a pA2 for [Nphe1]N/OFQ(1–13)NH2 (the template for UFP-101) against N/OFQ of 7.33±0.08 was obtained. Slope factors for the Schild regression lines were ~1 indicating competitivity. When UFP-101 is compared with its template molecule [Nphe1]N/OFQ(1–13)NH2, Arg14,Lys15 substitution produced ~1 log greater potency. We suggest that due to its high potency UFP-101 should prove a further useful tool in the evaluation of the N/OFQ-NOP receptor system.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2.
Fig. 3.

Similar content being viewed by others

References

  • Becker JA, Wallace A, Garzon A, Ingallinella P, Bianchi E, Cortese R, Simonin F, Kieffer BL, Pessi A (1999) Ligands for kappa-opioid and ORL1 receptors identified from a conformationally constrained peptide combinatorial library. J Biol Chem 274:27513–27522

    CAS  PubMed  Google Scholar 

  • Berger H, Calo G, Albrecht E, Guerrini R, Bienert M (2000) [Nphe1]NC(1–13)NH2 selectively antagonizes nociceptin/orphanin FQ-stimulated G-protein activation in rat brain. J Pharmacol Exp Ther 294:428–433

    CAS  PubMed  Google Scholar 

  • Bigoni R, Rizzi A, Rizzi D, Becker JA, Kieffer BL, Simonin F, Regoli D, Calo G (2000) In vitro pharmacological profile of peptide III-BTD: a novel ligand for nociceptin/orphanin FQ and opioid receptors. Life Sci 68:233–239

    Article  CAS  PubMed  Google Scholar 

  • Bigoni R, Calo G, Rizzi A, Okawa H, Regoli D, Smart D, Lambert DG (2002a) Effects of naloxone benzoylhydrazone on native and recombinant nociceptin/orphanin FQ receptors. Can J Physiol Pharmacol 80:407–412

    Article  CAS  PubMed  Google Scholar 

  • Bigoni R, Rizzi D, Rizzi A, Camarda V, Guerrini R, Lambert DG, Hashiba E, Berger H, Salvadori S, Regoli D, Calo G (2002b) Pharmacological characterization of [(pX)Phe4]nociceptin(1–13)amide analogs. I. In vitro studies. Naunyn-Schmiedebergs Arch Pharmacol 365:442–449

    Article  CAS  Google Scholar 

  • Calo G, Guerrini R, Bigoni R, Rizzi A, Marzola G, Okawa H, Bianchi C, Lambert DG, Salvadori S, Regoli D (2000a) Characterization of [Nphe1]nociceptin(1–13)NH2, a new selective nociceptin receptor antagonist. Br J Pharmacol 129:1183–1193

    CAS  PubMed  Google Scholar 

  • Calo G, Remo Guerrini, Rizzi A, Salvadri S, Regoli D (2000b) Pharmacology of nociceptin and its receptor: a novel therapeutic target. Br J Pharmacol 129:1261–1283

    CAS  PubMed  Google Scholar 

  • Calo G, Rizzi A, Rizzi D, Bigoni R, Guerrini R, Marzola G, Marti M, McDonald J, Morari M, Lambert DG, Salvadori S, Regoli D (2002) [Nphe1,Arg14,Lys15]Nociceptin-NH2, a novel potent and selective antagonist of the nociceptin/orphanin FQ receptor. Br J Pharmacol 136:303–311

    CAS  PubMed  Google Scholar 

  • Cheng Y, Prusoff WH (1973) Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction. Biochem Pharmacol 22:3099–3108

    CAS  PubMed  Google Scholar 

  • Cox BM, Chavkin C, Christie MJ, Civelli O, Evans C, Hamon MD, Hoellt V, Kieffer B, Kitchen I, McKnight AT, Meunier JC, Portoghese PS (2000) Opioid receptors. In: Girdlestone D (ed) The IUPHAR compendium of receptor characterization and classification. IUPHAR Media, London, pp 321–333

  • Guerrini R, Calo G, Rizzi A, Bianchi C, Lazarus LH, Salvadori S, Temussi PA, Regoli D (1997) Address and message sequences for the nociceptin receptor: a structure-activity study of nociceptin-(1–13)-peptide amide. J Med Chem 40:1789–1793

    CAS  PubMed  Google Scholar 

  • Guerrini R, Calo G, Bigoni R, Rizzi A, Varani K, Toth G, Gessi S, Hashiba E, Hashimoto Y, Lambert DG, Borea PA, Tomatis R, Salvadori S, Regoli D (2000a) Further studies on nociceptin-related peptides: discovery of a new chemical template with antagonist activity on the nociceptin receptor. J Med Chem 43:2805–2813

    CAS  PubMed  Google Scholar 

  • Guerrini R, Calo G, Rizzi A, Bigoni R, Rizzi D, Regoli D, Salvadori S (2000b) Structure-activity relationships of nociceptin and related peptides: comparison with dynorphin A. Peptides 21:923–933

    Article  CAS  PubMed  Google Scholar 

  • Guerrini R, Calo G, Bigoni R, Rizzi D, Regoli D, Salvadori S (2001a) Structure-activity relationship of [Nphe1]-NC-(1–13)-NH2, a pure and selective nociceptin/orphanin FQ receptor antagonist. J Pept Res 57:215–222

    Article  CAS  PubMed  Google Scholar 

  • Guerrini R, Calo G, Bigoni R, Rizzi D, Rizzi A, Zucchini M, Varani K, Hashiba E, Lambert DG, Toth G, Borea PA, Salvadori S, Regoli D (2001b) Structure-activity studies of the Phe4 residue of nociceptin(1–13)-NH2: identification of highly potent agonists of the nociceptin/orphanin FQ receptor. J Med Chem 44:3956–3964

    CAS  PubMed  Google Scholar 

  • Hashiba E, Harrison C, Galo G, Guerrini R, Rowbotham DJ, Smith G, Lambert DG (2001) Characterisation and comparison of novel ligands for the nociceptin/orphanin FQ receptor. Naunyn-Schmiedebergs Arch Pharmacol 363:28–33

    Article  CAS  Google Scholar 

  • Hashiba E, Lambert DG, Jenck F, Wichmann J, Smith G (2002) Characterisation of the non-peptide nociceptin receptor agonist, Ro64–6198 in Chinese hamster ovary cells expressing recombinant human nociceptin receptors. Life Sci 70:1719–1725

    Article  CAS  PubMed  Google Scholar 

  • Hashimoto Y, Calo G, Guerrini R, Smith G, Lambert DG (2000) Antagonistic effects of [Nphe1]nociceptin(1–13)NH2 on nociceptin receptor mediated inhibition of cAMP formation in Chinese hamster ovary cells stably expressing the recombinant human nociceptin receptor. Neurosci Lett 278:109–112

    CAS  PubMed  Google Scholar 

  • Ichikawa D, Ozaki S, Azuma T, Nambu H, Kawamoto H, Iwasawa Y, Takeshima H, Ohta H (2001) In vitro inhibitory effects of J-113397 on nociceptin/orphanin FQ-stimulated. Neuroreport 12:1757–1761

    CAS  PubMed  Google Scholar 

  • Kobayashi T, Ikeda K, Togashi S, Itoh N, Kumanishi T (1997) Effects of sigma ligands on the nociceptin/orphanin FQ receptor co-expressed with the G-protein-activated K+ channel in Xenopus oocytes. Br J Pharmacol 120:986–987

    CAS  PubMed  Google Scholar 

  • McDonald J, Barnes TA, Calo G, Guerrini R, Rowbotham DJ, Lambert DG (2001) In vitro characterisation of [(pF)Phe4]nociceptin(1–13)NH2 a novel agonist for the NOP receptor. Br J Pharmacol [Suppl]:258P

  • McDonald J, Barnes TA, Calo G, Guerrini R, Rowbotham DJ, Lambert DG (2002) Effects of [(pF)Phe4]nociceptin/orphanin FQ-(1–13)NH2 on GTPgamma35S binding and cAMP formation in Chinese hamster ovary cells expressing the human nociceptin/orphanin FQ receptor. Eur J Pharmacol 443:7–12

    Article  CAS  PubMed  Google Scholar 

  • Meunier JC, Mollereau C, Toll L, Suaudeau C, Moisand C, Alvinerie P, Butour JL, Guillemot JC, Ferrara P, Monsarrat B, Mazaguil H, Vassart G, Permentier M, Costentin J (1995) Isolation and structure of the endogenous agonist of opioid receptor-like ORL1 receptor. Nature 377:532–535

    CAS  PubMed  Google Scholar 

  • Mogil JS, Pasternak GW (2001) The molecular and behavioral pharmacology of the orphanin FQ/nociceptin peptide and receptor family. Pharmacol Rev 53:381–415

    CAS  PubMed  Google Scholar 

  • Okada K, Sujaku T, Chuman Y, Nakashima R, Nose T, Costa T, Yamada Y, Yokoyama M, Nagahisa A, Shimohigashi Y (2000) Highly potent nociceptin analog containing the Arg-Lys triple repeat. Biochem Biophys Res Commun 278:493–498

    CAS  PubMed  Google Scholar 

  • Ozaki S, Kawamoto H, Itoh Y, Miyaji M, Iwasawa Y, Ohta H (2000) A potent and highly selective nonpeptidyl nociceptin/orphanin FQ receptor (ORL1) antagonist: J-113397. Eur J Pharmacol 387:R17–18

    CAS  PubMed  Google Scholar 

  • Reinscheid RK, Nothacker HP, Bourson A, Ardati A, Henningsen RA, Bunzow JR, Grandy DK, Langen H, Monsma FJ Jr, Civelli O (1995) Orphanin FQ: a neuropeptide that activates an opioid-like G protein-coupled receptor. Science 270:792–794

    CAS  PubMed  Google Scholar 

  • Rizzi D, Rizzi A, Bigoni R, Camarda V, Marzola G, Guerrini R, De Risi C, Regoli D Calo G (2002) [Arg14,Lys15]nociceptin, a highly potent agonist of the nociceptin/orphanin FQ receptor: in vitro and in vivo studies. J Pharmacol Exp Ther 300:57–63

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We would like to thank Drs F. Jenck and J. Wichmann of CNS Research, Roche Pharma Division, Basel, Switzerland for provision of Ro64–6198.

Author information

Authors and Affiliations

  1. University Department of Anaesthesia, Critical Care and Pain Management, Leicester Royal Infirmary, Leicester, LE1 5WW, UK

    J. McDonald & D. G. Lambert

  2. Department of Experimental and Clinical Medicine, Section of Pharmacology and Neuroscience Center, University of Ferrara, via Fossato di Mortara 17, 44100, Ferrara, Italy

    G. Calo

  3. Department of Pharmaceutical Sciences and Biotechnology Center, University of Ferrara, via Fossato di Mortara 17, 44100, Ferrara, Italy

    R. Guerrini

Authors
  1. J. McDonald
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. Calo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R. Guerrini
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. G. Lambert
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to D. G. Lambert.

Rights and permissions

Reprints and permissions

About this article

Cite this article

McDonald, J., Calo, G., Guerrini, R. et al. UFP-101, a high affinity antagonist for the nociceptin/orphanin FQ receptor: radioligand and GTPγ35S binding studies. Naunyn-Schmiedeberg's Arch Pharmacol 367, 183–187 (2003). https://doi.org/10.1007/s00210-002-0661-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00210-002-0661-8

Keywords

Search

Navigation