Tritium-labelled isovaleryl-RYYRIK-NH2 as potential antagonist probe for ORL1 nociceptin receptor

Shogo Inamine, Hirokazu Nishimura, Jinglan Li, Kaname Isozaki, Ayami Matsushima, Tommaso Costa, Yasuyuki Shimohigashi

研究成果: ジャーナルへの寄稿記事

2 引用 (Scopus)

抄録

IsoVa-RYYRIK-NH2 is a highly specific antagonist ligand of the opioid receptor-like 1 (ORL1) receptor, an endogenous ligand of which is 17-mer peptide nociceptin. ORL1 antagonists have potential for clinical use as analgesic and antineuropathic drugs, and thus information on the receptor-binding characteristics of antagonists is very important for rational drug design. In the present study, we prepared tritium-labelled isova-RYYRIK-NH2 from its precursor with the 3-methylcrotonyl (CH3)2CCHCO group by a catalytic reduction using tritium gas. The resulting [3H]isoVa-RYYRIK-NH2 was evaluated in a saturation binding assay using the COS-7 cell membrane preparations of transiently expressed ORL1. It exhibited more than 90% specific binding with a dissociation constant of 1.21 ± 0.03 nM. From the mutual heterologous binding assays using [3H]isoVa-RYYRIK-NH2 and [3H]nociceptin, isoVa-RYYRIK-NH2 and nociceptin were found to share the receptor-binding site, but each also had a separate specific binding site of its own. They differentiated the two different binding states or conformations of ORL1, which might represent the agonist-active and antagonist-inactive conformations of ORL1. [3H]isoVa-RYYRIK-NH2 is thus a key tracer to uncover the amino acid residues important for receptor inactivation.

元の言語英語
ページ(範囲)5902-5909
ページ数8
ジャーナルBioorganic and Medicinal Chemistry
22
発行部数21
DOI
出版物ステータス出版済み - 11 1 2014

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Narcotic Antagonists
Tritium
Opioid Receptors
Binding Sites
Conformations
Ligands
Assays
Drug Design
COS Cells
Analgesics
Cell membranes
Gases
Cell Membrane
Amino Acids
Peptides
nociceptin receptor
nociceptin
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

これを引用

Tritium-labelled isovaleryl-RYYRIK-NH2 as potential antagonist probe for ORL1 nociceptin receptor. / Inamine, Shogo; Nishimura, Hirokazu; Li, Jinglan; Isozaki, Kaname; Matsushima, Ayami; Costa, Tommaso; Shimohigashi, Yasuyuki.

:: Bioorganic and Medicinal Chemistry, 巻 22, 番号 21, 01.11.2014, p. 5902-5909.

研究成果: ジャーナルへの寄稿記事

Inamine, Shogo ; Nishimura, Hirokazu ; Li, Jinglan ; Isozaki, Kaname ; Matsushima, Ayami ; Costa, Tommaso ; Shimohigashi, Yasuyuki. / Tritium-labelled isovaleryl-RYYRIK-NH2 as potential antagonist probe for ORL1 nociceptin receptor. :: Bioorganic and Medicinal Chemistry. 2014 ; 巻 22, 番号 21. pp. 5902-5909.
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abstract = "IsoVa-RYYRIK-NH2 is a highly specific antagonist ligand of the opioid receptor-like 1 (ORL1) receptor, an endogenous ligand of which is 17-mer peptide nociceptin. ORL1 antagonists have potential for clinical use as analgesic and antineuropathic drugs, and thus information on the receptor-binding characteristics of antagonists is very important for rational drug design. In the present study, we prepared tritium-labelled isova-RYYRIK-NH2 from its precursor with the 3-methylcrotonyl (CH3)2CCHCO group by a catalytic reduction using tritium gas. The resulting [3H]isoVa-RYYRIK-NH2 was evaluated in a saturation binding assay using the COS-7 cell membrane preparations of transiently expressed ORL1. It exhibited more than 90{\%} specific binding with a dissociation constant of 1.21 ± 0.03 nM. From the mutual heterologous binding assays using [3H]isoVa-RYYRIK-NH2 and [3H]nociceptin, isoVa-RYYRIK-NH2 and nociceptin were found to share the receptor-binding site, but each also had a separate specific binding site of its own. They differentiated the two different binding states or conformations of ORL1, which might represent the agonist-active and antagonist-inactive conformations of ORL1. [3H]isoVa-RYYRIK-NH2 is thus a key tracer to uncover the amino acid residues important for receptor inactivation.",
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AU - Inamine, Shogo

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AU - Li, Jinglan

AU - Isozaki, Kaname

AU - Matsushima, Ayami

AU - Costa, Tommaso

AU - Shimohigashi, Yasuyuki

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AB - IsoVa-RYYRIK-NH2 is a highly specific antagonist ligand of the opioid receptor-like 1 (ORL1) receptor, an endogenous ligand of which is 17-mer peptide nociceptin. ORL1 antagonists have potential for clinical use as analgesic and antineuropathic drugs, and thus information on the receptor-binding characteristics of antagonists is very important for rational drug design. In the present study, we prepared tritium-labelled isova-RYYRIK-NH2 from its precursor with the 3-methylcrotonyl (CH3)2CCHCO group by a catalytic reduction using tritium gas. The resulting [3H]isoVa-RYYRIK-NH2 was evaluated in a saturation binding assay using the COS-7 cell membrane preparations of transiently expressed ORL1. It exhibited more than 90% specific binding with a dissociation constant of 1.21 ± 0.03 nM. From the mutual heterologous binding assays using [3H]isoVa-RYYRIK-NH2 and [3H]nociceptin, isoVa-RYYRIK-NH2 and nociceptin were found to share the receptor-binding site, but each also had a separate specific binding site of its own. They differentiated the two different binding states or conformations of ORL1, which might represent the agonist-active and antagonist-inactive conformations of ORL1. [3H]isoVa-RYYRIK-NH2 is thus a key tracer to uncover the amino acid residues important for receptor inactivation.

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