Fluorescent-labeled bioconjugates of the opioid peptides biphalin and DPDPE incorporating fluorescein-maleimide linkers

Azzurra Stefanucci, Wei Lei, Victor J Hruby, Giorgia Macedonio, Grazia Luisi, Simone Carradori, John M. Streicher, Adriano Mollica

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Aim: The conjugation of fluorescent labels to opioid peptides is an extremely challenging task, which needs to be overcome to create new classes of probes for biological assays. Materials & methods: Three opioid peptide analogs of biphalin and [D-Pen2,5]-Enkephalin (DPDPE) containing a fluorescein-maleimide motif were synthesized. Results & discussion: The biphalin analog 17 binds to opioid receptors with Ki μ = 530 ± 90 nM and Ki δ = 69.8 ± 16.4 nM. We then tested the ability of the compounds to stimulate G-protein-coupling, 17 activated μ-receptor expressing cells (EC50 = 16.7 ± 6.7 nM, EMax = 76 ± 4%) as well as δ-receptor expressing cells (EC50 = 42 ± 10 nM, EMax = 34 ± 8%). However, 17 was not able to fluorescently label receptor in live or fixed cells. Conclusion: Our data suggest that the biphalin scaffold could be employed to develop fluorescent ligands with the appropriate fluorescent motif, and suggest a means for further probe development.

Original languageEnglish (US)
Pages (from-to)859-869
Number of pages11
JournalFuture Medicinal Chemistry
Volume9
Issue number9
DOIs
StatePublished - Jun 1 2017

Fingerprint

D-Penicillamine (2,5)-Enkephalin
Opioid Peptides
Enkephalins
Opioid Receptors
GTP-Binding Proteins
Biological Assay
Ligands
fluorescein 5-maleimide
biphalin

Keywords

  • biphalin
  • DPDPE
  • fluorescein
  • fluorescent probes
  • GPCRs
  • opioid peptides

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology
  • Drug Discovery

Cite this

Fluorescent-labeled bioconjugates of the opioid peptides biphalin and DPDPE incorporating fluorescein-maleimide linkers. / Stefanucci, Azzurra; Lei, Wei; Hruby, Victor J; Macedonio, Giorgia; Luisi, Grazia; Carradori, Simone; Streicher, John M.; Mollica, Adriano.

In: Future Medicinal Chemistry, Vol. 9, No. 9, 01.06.2017, p. 859-869.

Research output: Contribution to journalArticle

Stefanucci, Azzurra ; Lei, Wei ; Hruby, Victor J ; Macedonio, Giorgia ; Luisi, Grazia ; Carradori, Simone ; Streicher, John M. ; Mollica, Adriano. / Fluorescent-labeled bioconjugates of the opioid peptides biphalin and DPDPE incorporating fluorescein-maleimide linkers. In: Future Medicinal Chemistry. 2017 ; Vol. 9, No. 9. pp. 859-869.
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AU - Streicher, John M.

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