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

doi:10.4155/fmc-2016-0232

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 journal › Article › peer-review

25 Scopus citations

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 K_i ^μ = 530 ± 90 nM and K_i ^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, E_Max = 76 ± 4%) as well as δ-receptor expressing cells (EC₅₀ = 42 ± 10 nM, E_Max = 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 language	English (US)
Pages (from-to)	859-869
Number of pages	11
Journal	Future Medicinal Chemistry
Volume	9
Issue number	9
DOIs	https://doi.org/10.4155/fmc-2016-0232
State	Published - Jun 2017

Keywords

DPDPE
GPCRs
biphalin
fluorescein
fluorescent probes
opioid peptides

ASJC Scopus subject areas

Molecular Medicine
Pharmacology
Drug Discovery

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title = "Fluorescent-labeled bioconjugates of the opioid peptides biphalin and DPDPE incorporating fluorescein-maleimide linkers",

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.",

keywords = "DPDPE, GPCRs, biphalin, fluorescein, fluorescent probes, opioid peptides",

author = "Azzurra Stefanucci and Wei Lei and Hruby, {Victor J.} and Giorgia Macedonio and Grazia Luisi and Simone Carradori and Streicher, {John M.} and Adriano Mollica",

note = "Publisher Copyright: {\textcopyright} 2017 Future Science Ltd.",

year = "2017",

month = jun,

doi = "10.4155/fmc-2016-0232",

language = "English (US)",

volume = "9",

pages = "859--869",

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T1 - Fluorescent-labeled bioconjugates of the opioid peptides biphalin and DPDPE incorporating fluorescein-maleimide linkers

AU - Stefanucci, Azzurra

AU - Lei, Wei

AU - Hruby, Victor J.

AU - Macedonio, Giorgia

AU - Luisi, Grazia

AU - Carradori, Simone

AU - Streicher, John M.

AU - Mollica, Adriano

PY - 2017/6

Y1 - 2017/6

N2 - 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.

AB - 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.

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

Abstract

Keywords

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