In Vitro Membrane Permeation Studies and in Vivo Antinociception of Glycosylated Dmt1-DALDA Analogues

Steven Ballet, Cecilia Betti, Alexandre Novoa, Csaba Tömböly, Carsten Uhd Nielsen, Hans Christian Helms, Anna Lesniak, Patrycja Kleczkowska, Nga N Chung, Andrzej W Lipkowski, Birger Brodin, Dirk Tourwé, Peter W Schiller

Publikation: Bidrag til tidsskriftLetterForskningpeer review

Resumé

In this study the μ opioid receptor (MOR) ligands DALDA (Tyr-d-Arg-Phe-Lys-NH2) and Dmt(1)-DALDA (Dmt-d-Arg-Phe-Lys-NH2, Dmt = 2',6'-dimethyltyrosine) were glycosylated at the N- or C-terminus. Subsequently, the modified peptides were subjected to in vitro and in vivo evaluation. In contrast to the N-terminally modified peptide (3), all peptide analogues derivatized at the C-terminus (4-7) proved to possess high affinity and agonist potency at both MOR and DOR (δ opioid receptor). Results of the Caco-2 monolayer permeation, as well as in vitro blood-brain barrier model experiments, showed that, in the case of compound 4, the glycosylation only slightly diminished the lumen-to-blood and blood-to-lumen transport. Altogether, these experiments were indicative of transcellular transport but not active transport. In vivo assays demonstrated that the peptides were capable of (i) crossing the blood-brain barrier (BBB) and (ii) activating both the spinal ascending as well as the descending opioid pathways, as determined by the tail-flick and hot-plate assays, respectively. In contrast to the highly selective MOR agonist Dmt(1)-DALDA 1, compounds 4-7 are mixed MOR/DOR agonists, expected to produce reduced opioid-related side effects.

OriginalsprogEngelsk
TidsskriftA C S Medicinal Chemistry Letters
Vol/bind5
Udgave nummer4
Sider (fra-til)352-357
ISSN1948-5875
DOI
StatusUdgivet - 10. apr. 2014
Udgivet eksterntJa

Fingeraftryk

Permeation
Membranes
Peptides
Opioid Receptors
Opioid Analgesics
Assays
Blood
Glycosylation
Monolayers
Experiments
Ligands
tyrosyl-arginyl-phenylalanyl-lysinamide
In Vitro Techniques
Blood-Brain Barrier
2',6'-dimethyltyrosyl-arginyl-phenylalanyl-lysinamide

Citer dette

Ballet, Steven ; Betti, Cecilia ; Novoa, Alexandre ; Tömböly, Csaba ; Nielsen, Carsten Uhd ; Helms, Hans Christian ; Lesniak, Anna ; Kleczkowska, Patrycja ; Chung, Nga N ; Lipkowski, Andrzej W ; Brodin, Birger ; Tourwé, Dirk ; Schiller, Peter W. / In Vitro Membrane Permeation Studies and in Vivo Antinociception of Glycosylated Dmt1-DALDA Analogues. I: A C S Medicinal Chemistry Letters. 2014 ; Bind 5, Nr. 4. s. 352-357.
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title = "In Vitro Membrane Permeation Studies and in Vivo Antinociception of Glycosylated Dmt1-DALDA Analogues",
abstract = "In this study the μ opioid receptor (MOR) ligands DALDA (Tyr-d-Arg-Phe-Lys-NH2) and Dmt(1)-DALDA (Dmt-d-Arg-Phe-Lys-NH2, Dmt = 2',6'-dimethyltyrosine) were glycosylated at the N- or C-terminus. Subsequently, the modified peptides were subjected to in vitro and in vivo evaluation. In contrast to the N-terminally modified peptide (3), all peptide analogues derivatized at the C-terminus (4-7) proved to possess high affinity and agonist potency at both MOR and DOR (δ opioid receptor). Results of the Caco-2 monolayer permeation, as well as in vitro blood-brain barrier model experiments, showed that, in the case of compound 4, the glycosylation only slightly diminished the lumen-to-blood and blood-to-lumen transport. Altogether, these experiments were indicative of transcellular transport but not active transport. In vivo assays demonstrated that the peptides were capable of (i) crossing the blood-brain barrier (BBB) and (ii) activating both the spinal ascending as well as the descending opioid pathways, as determined by the tail-flick and hot-plate assays, respectively. In contrast to the highly selective MOR agonist Dmt(1)-DALDA 1, compounds 4-7 are mixed MOR/DOR agonists, expected to produce reduced opioid-related side effects.",
author = "Steven Ballet and Cecilia Betti and Alexandre Novoa and Csaba T{\"o}mb{\"o}ly and Nielsen, {Carsten Uhd} and Helms, {Hans Christian} and Anna Lesniak and Patrycja Kleczkowska and Chung, {Nga N} and Lipkowski, {Andrzej W} and Birger Brodin and Dirk Tourw{\'e} and Schiller, {Peter W}",
year = "2014",
month = "4",
day = "10",
doi = "10.1021/ml4004765",
language = "English",
volume = "5",
pages = "352--357",
journal = "A C S Medicinal Chemistry Letters",
issn = "1948-5875",
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Ballet, S, Betti, C, Novoa, A, Tömböly, C, Nielsen, CU, Helms, HC, Lesniak, A, Kleczkowska, P, Chung, NN, Lipkowski, AW, Brodin, B, Tourwé, D & Schiller, PW 2014, 'In Vitro Membrane Permeation Studies and in Vivo Antinociception of Glycosylated Dmt1-DALDA Analogues', A C S Medicinal Chemistry Letters, bind 5, nr. 4, s. 352-357. https://doi.org/10.1021/ml4004765

In Vitro Membrane Permeation Studies and in Vivo Antinociception of Glycosylated Dmt1-DALDA Analogues. / Ballet, Steven; Betti, Cecilia; Novoa, Alexandre; Tömböly, Csaba; Nielsen, Carsten Uhd; Helms, Hans Christian; Lesniak, Anna; Kleczkowska, Patrycja; Chung, Nga N; Lipkowski, Andrzej W; Brodin, Birger; Tourwé, Dirk; Schiller, Peter W.

I: A C S Medicinal Chemistry Letters, Bind 5, Nr. 4, 10.04.2014, s. 352-357.

Publikation: Bidrag til tidsskriftLetterForskningpeer review

TY - JOUR

T1 - In Vitro Membrane Permeation Studies and in Vivo Antinociception of Glycosylated Dmt1-DALDA Analogues

AU - Ballet, Steven

AU - Betti, Cecilia

AU - Novoa, Alexandre

AU - Tömböly, Csaba

AU - Nielsen, Carsten Uhd

AU - Helms, Hans Christian

AU - Lesniak, Anna

AU - Kleczkowska, Patrycja

AU - Chung, Nga N

AU - Lipkowski, Andrzej W

AU - Brodin, Birger

AU - Tourwé, Dirk

AU - Schiller, Peter W

PY - 2014/4/10

Y1 - 2014/4/10

N2 - In this study the μ opioid receptor (MOR) ligands DALDA (Tyr-d-Arg-Phe-Lys-NH2) and Dmt(1)-DALDA (Dmt-d-Arg-Phe-Lys-NH2, Dmt = 2',6'-dimethyltyrosine) were glycosylated at the N- or C-terminus. Subsequently, the modified peptides were subjected to in vitro and in vivo evaluation. In contrast to the N-terminally modified peptide (3), all peptide analogues derivatized at the C-terminus (4-7) proved to possess high affinity and agonist potency at both MOR and DOR (δ opioid receptor). Results of the Caco-2 monolayer permeation, as well as in vitro blood-brain barrier model experiments, showed that, in the case of compound 4, the glycosylation only slightly diminished the lumen-to-blood and blood-to-lumen transport. Altogether, these experiments were indicative of transcellular transport but not active transport. In vivo assays demonstrated that the peptides were capable of (i) crossing the blood-brain barrier (BBB) and (ii) activating both the spinal ascending as well as the descending opioid pathways, as determined by the tail-flick and hot-plate assays, respectively. In contrast to the highly selective MOR agonist Dmt(1)-DALDA 1, compounds 4-7 are mixed MOR/DOR agonists, expected to produce reduced opioid-related side effects.

AB - In this study the μ opioid receptor (MOR) ligands DALDA (Tyr-d-Arg-Phe-Lys-NH2) and Dmt(1)-DALDA (Dmt-d-Arg-Phe-Lys-NH2, Dmt = 2',6'-dimethyltyrosine) were glycosylated at the N- or C-terminus. Subsequently, the modified peptides were subjected to in vitro and in vivo evaluation. In contrast to the N-terminally modified peptide (3), all peptide analogues derivatized at the C-terminus (4-7) proved to possess high affinity and agonist potency at both MOR and DOR (δ opioid receptor). Results of the Caco-2 monolayer permeation, as well as in vitro blood-brain barrier model experiments, showed that, in the case of compound 4, the glycosylation only slightly diminished the lumen-to-blood and blood-to-lumen transport. Altogether, these experiments were indicative of transcellular transport but not active transport. In vivo assays demonstrated that the peptides were capable of (i) crossing the blood-brain barrier (BBB) and (ii) activating both the spinal ascending as well as the descending opioid pathways, as determined by the tail-flick and hot-plate assays, respectively. In contrast to the highly selective MOR agonist Dmt(1)-DALDA 1, compounds 4-7 are mixed MOR/DOR agonists, expected to produce reduced opioid-related side effects.

U2 - 10.1021/ml4004765

DO - 10.1021/ml4004765

M3 - Letter

C2 - 24839540

VL - 5

SP - 352

EP - 357

JO - A C S Medicinal Chemistry Letters

JF - A C S Medicinal Chemistry Letters

SN - 1948-5875

IS - 4

ER -