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δ Opioidmimetic Antagonists: Prototypes for Designing a New Generation of Ultraselective Opioid Peptides

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Abstract

Background

Tyr-Tic (1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid) and Tyr-Tic-Ala were the first peptides with δ opioid antagonist activity lacking Phe, considered essential for opioid activity based on the N-terminal tripeptide sequence (Tyr-d-Xaa-Phe) of amphibian skin opioids. Analogs were then designed to restrain the rotational flexibility of Tyr by the substitution of 2,6-dimethyl-l-tyrosine (Dmt).

Materials and Methods

Tyr and Dmt peptides were synthesized by solid phase and solution methods using Fmoc technology or condensing Boc-Dmt-OH or Boc-Tyr(But)-OH with H-l-Tic-OBut or H-d-Tic-OBut, respectively. Peptides were purified (>99%) by HPLC and characteristics determined by 1H-NMR, FAB-MS, melting point, TLC, and amino acid analyses.

Results

H-Dmt-Tic-OH had high affinity (Kiδ = 0.022 nM) and extraordinary selectivity (Kiμ/Kiδ = 150,000); H-Dmt-Tic-Ala-OH had a Kiδ = 0.29 nM and δ selectivity = 20,000. Affinity and selectivity increased 8700- and 1000-fold relative to H-Tyr-Tic-OH, respectively. H-Dmt-Tic-OH and H-Dmt-Tic-NH2 fitted one-site receptor binding models (η = 0.939−0.987), while H-Dmt-Tic-ol, H-Dmt-Tic-Ala-OH and H-Dmt-Tic-Ala-NH2 best fitted two-site models (η = 0.708−0.801, F 18.9−26.0, p < 0.0001). Amidation increased μ affinity by 10- to 100-fold and acted synergistically with d-Tic2 to reverse selectivity (δ → μ). Dmt-Tic di- and tripeptides exhibited δ antagonist bioactivity (Ke = 4−66 nM) with mouse vas deferens and lacked agonist μ activity (> 10 µM) in guinea-pig ileum preparations. Dmt-Tic analogs weakly interacted with κ receptors in the 1 to >20 µM range.

Conclusions

Dmt-Tic opioidmimetic peptides represent a highly potent class of opioid peptide antagonists with greater potency than the nonopioid δ antagonist naltrindole and have potential application as clinical and therapeutic compounds.

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Acknowledgments

We are indebted to Dr. J. H. Dygos (G. D. Searle and Co.) for generously providing us with 2′,6′-dimethyl-l-tyrosine. We thank Dr. R. Anacardio, Dompé SpA L’Aquila-Italy for the FAB-mass spectroscopic determinations. We appreciate the part-time assistance of K. Jefferies and T. McElwee, and the consistent library support by S. Fuller, R. J. Hester, E. M. Leadem, and F. T. Lyndon. We are grateful to Dr. C. Gruppi at Morgan & Finnegan, Attorneys at Law, New York, NY, for studiously preparing and submitting our data for a patent application, Serial Number 08/347.531, through the aegis of the National Institutes of Health Office of Technology Transfer. S. Salvadori was supported in part by grants from CNR Progetto Finalizzato Chimica Fine e Secondaria II and Murst.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Ferrara, Ferrara, Italy

    Severo Salvadori, Giofranco Balboni & Remo Guerrini

  2. Institute of Pharmacology, University of Ferrara, Ferrara, Italy

    Clementina Bianchi

  3. Department of Pharmacy, Division of Pharmacology and Toxicology, University of Helsinki, Helsinki, Finland

    Martti Attila

  4. Peptide Neurochemistry, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, USA

    Sharon D. Bryant & Lawrence H. Lazarus

  5. Department of Chemistry, University of Naples Federico II, Naples, Italy

    Orlando Crescenzi, Delia Picone & Piero A. Temussi

  6. Institute of Chemistry, MIB del CNR, Naples, Italy

    Teodorico Tancredi

  7. NIEHS, P.O. Box 12233, Research Triangle Park, North Carolina, 27709, USA

    Lawrence H. Lazarus

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  1. Severo Salvadori
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Salvadori, S., Attila, M., Balboni, G. et al. δ Opioidmimetic Antagonists: Prototypes for Designing a New Generation of Ultraselective Opioid Peptides. Mol Med 1, 678–689 (1995). https://doi.org/10.1007/BF03401608

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