Synthesis and Evaluation of Derivatives of Leucine Enkephalin as Potential Affinity Labels for δ Opioid Receptors

Heekyung Choi, Thomas F. Murray, Jane V. Aldrich

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

As part of an effort to develop peptide-based affinity labels for opioid receptors, [Leu5]enkephalin (LeuEnk) and DTLET (Tyr-D-Thr-Gly-Phe-Leu-Thr), potent agonists for δ receptors, were selected as the parent peptides for further modification. The affinity label derivatives were prepared using standard Fmoc solid-phase peptide synthesis in conjunction with Fmoc-Phe(p-NHAlloc) (Fmoc: 9-flourenylmethoxycarbonyl;) and selective modification of the p-amino group on this residue. The electrophilic isothiocyanate and bromoacetamide groups were introduced into the para position of Phe4; the corresponding free amine-containing peptides were also prepared for comparison. The pure peptides were evaluated in radioligand binding assays using Chinese hamster ovary (CHO) cells expressing δ and μ opioid receptors. Modification of Phe4 in LeuEnk and DTLET significantly decreased δ-receptor binding affinity (40 to >2,000-fold). Among the synthesized analogues, [Phe(p-NH2) 4]DTLET showed the highest δ-receptor binding affinity (IC 50 = 39 nM) and enhanced selectivity for δ receptors compared to DTLET while other derivatives exhibited much lower δ receptor affinity. The differences in affinities between the two series of analogues and between the derivatives of LeuEnk and N,N-dibenzyl[Leu5]Enk reported previously suggest subtle differences in interactions of Phe4 with δ receptors depending on other modifications in the sequences.

Original languageEnglish
Pages (from-to)552-557
Number of pages6
JournalBiopolymers
Volume71
Issue number5
DOIs
StatePublished - 2003
Externally publishedYes

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Affinity Labels
Leucine Enkephalin
Opioid Receptors
Peptides
Labels
Enkephalins
Derivatives
Radioligand Assay
Solid-Phase Synthesis Techniques
Cricetulus
Amines
Ovary
Assays
Cells
deltakephalin

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics

Cite this

Synthesis and Evaluation of Derivatives of Leucine Enkephalin as Potential Affinity Labels for δ Opioid Receptors. / Choi, Heekyung; Murray, Thomas F.; Aldrich, Jane V.

In: Biopolymers, Vol. 71, No. 5, 2003, p. 552-557.

Research output: Contribution to journalArticle

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