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Pharmacological profile of the cyclic nociceptin/orphanin FQ analogues c[Cys10,14]N/OFQ(1–14)NH2 and c[Nphe1,Cys10,14]N/OFQ(1–14)NH2

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Abstract

In this study we describe the activity of two cyclic nociceptin/orphanin FQ (N/OFQ) peptides; c[Cys10,14]N/OFQ(1–14)NH2 (c[Cys10,14]) and its [Nphe1] derivative c[Nphe1,Cys10,14]N/OFQ(1–14)NH2 (c[Nphe1,Cys10,14]) in native rat and mouse and recombinant human N/OFQ receptors (NOP). Cyclisation may protect the peptide from metabolic degradation.

In competition binding studies of rat, mouse and human NOP the following rank order pKi was obtained: N/OFQ(1–13)NH2(reference agonist)>N/OFQ=c[Cys10,14]>>c[Nphe1Cys10,14]. In GTPγ35S studies of Chinese hamster ovary cells expressing human NOP (CHOhNOP) c[Cys10,14] (pEC50 8.29) and N/OFQ(1–13)NH2 (pEC50 8.57) were full agonists whilst c[Nphe1Cys10,14] alone was inactive. Following 30 min pre-incubation c[Nphe1Cys10,14] competitively antagonised the effects of N/OFQ(1–13)NH2 with a pA2 and slope factor of 6.92 and 1.01 respectively. In cAMP assays c[Cys10,14] (pEC50 9.29, Emax 102% inhibition of the forskolin stimulated response), N/OFQ(1–13)NH2 (pEC50 10.16, Emax 103% inhibition) and c[Nphe1Cys10,14] (~80% inhibition at 10 μM) displayed agonist activity. In the mouse vas deferens c[Cys10,14] (pEC50 6.82, Emax 89% inhibition of electrically evoked contractions) and N/OFQ(1–13)NH2 (pEC50 7.47, Emax 93% inhibition) were full agonists whilst c[Nphe1Cys10,14] alone was inactive. c[Nphe1Cys10,14] (10 μM) competitively antagonised the effects of N/OFQ(1–13)NH2 with a pKB of 5.66. In a crude attempt to assess metabolic stability, c[Cys10,14] was incubated with rat brain membranes and then the supernatant assayed for remaining peptide. Following 60 min incubation 64% of the 1 nM added peptide was metabolised (compared with 54% for N/OFQ-NH2).

In summary, we report that c[Cys10,14] is a full agonist with a small reduction in potency but no improvement in stability whilst c[Nphe1Cys10,14] displays tissue (antagonist in the vas deferens) and assay (antagonist in the GTPγ35S assay and agonist in cAMP assay) dependent activity.

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Acknowledgement

The authors would like to thank the International Association for the Study of Pain for provision of a collaborative travel grant between Leicester (UK) and Ferrara (Italy).

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Authors and Affiliations

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

    M. Kitayama, T. A. Barnes, J. McDonald, D. J. Rowbotham, G. Smith & D. G. Lambert

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

    G. Carra & G. Calo

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

    R. Guerrini

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  1. M. Kitayama
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  2. T. A. Barnes
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  3. G. Carra
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  4. J. McDonald
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  5. G. Calo
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  7. D. J. Rowbotham
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  8. G. Smith
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  9. D. G. Lambert
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Correspondence to D. G. Lambert.

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Presented in part to The British Pharmacological Society at the Brighton, UK Meeting January 2003

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Kitayama, M., Barnes, T.A., Carra, G. et al. Pharmacological profile of the cyclic nociceptin/orphanin FQ analogues c[Cys10,14]N/OFQ(1–14)NH2 and c[Nphe1,Cys10,14]N/OFQ(1–14)NH2 . Naunyn-Schmiedeberg's Arch Pharmacol 368, 528–537 (2003). https://doi.org/10.1007/s00210-003-0821-5

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