Synthesis and hyperpolarisation of eNOS substrates for quantification of NO production by (1)H NMR spectroscopy

Fernando Fernandez Diaz-Rullo, Francesco Zamberlan, Ryan E. Mewis, Marianna Fekete, Lionel Broche, Lesley A. Cheyne, Sergio Dall'Angelo, Simon B. Duckett, Dana Dawson, Matteo Zanda

Research output: Contribution to journalArticle

2 Citations (Scopus)
3 Downloads (Pure)

Abstract

Hyperpolarization enhances the intensity of the NMR signals of a molecule, whose in vivo metabolic fate can be monitored by MRI with higher sensitivity. SABRE is a hyperpolarization technique that could potentially be used to image nitric oxide (NO) production in vivo. This would be very important, because NO dysregulation is involved in several pathologies, including cardiovascular ones. The nitric oxide synthase (NOS) pathway leads to NO production via conversion of L-arginine into L-citrulline. NO is a free radical gas with a short half-life in vivo (≈5 s), therefore direct NO quantification is challenging. An indirect method – based on quantifying conversion of an L-Arg- to L-Cit-derivative by 1H NMR spectroscopy – is herein proposed. A small library of pyridyl containing L-Arg derivatives was designed and synthesised. In vitro tests showed that compounds 4a–j and 11a–c were better or equivalent substrates for the eNOS enzyme (NO2 production = 19–46 μM) than native L-Arg (NO2 production = 25 μM). Enzymatic conversion of L-Arg to L-Cit derivatives could be monitored by 1H NMR. The maximum hyperpolarization achieved by SABRE reached 870-fold NMR signal enhancement, which opens up exciting future perspectives of using these molecules as hyperpolarized MRI tracers in vivo.

Original languageEnglish
Pages (from-to)2730-2742
Number of pages13
JournalBioorganic & Medicinal Chemistry
Volume25
Issue number10
Early online date21 Mar 2017
DOIs
Publication statusPublished - 15 May 2017

Fingerprint

Nitric Oxide
Magnetic Resonance Spectroscopy
Citrulline
Nitric Oxide Synthase
Libraries
Free Radicals
Half-Life
Arginine
Gases
Pathology
Enzymes
Proton Magnetic Resonance Spectroscopy

Keywords

  • Animals
  • Arginine
  • Biocatalysis
  • Cattle
  • Magnetic Resonance Spectroscopy
  • Nitric Oxide
  • Nitric Oxide Synthase Type III
  • Recombinant Proteins
  • Substrate Specificity
  • Journal Article
  • Hyperpolarization
  • L-Arginine
  • MRI
  • Real-time imaging
  • SABRE

Cite this

Synthesis and hyperpolarisation of eNOS substrates for quantification of NO production by (1)H NMR spectroscopy. / Fernandez Diaz-Rullo, Fernando; Zamberlan, Francesco; Mewis, Ryan E.; Fekete, Marianna; Broche, Lionel; Cheyne, Lesley A.; Dall'Angelo, Sergio; Duckett, Simon B.; Dawson, Dana; Zanda, Matteo.

In: Bioorganic & Medicinal Chemistry, Vol. 25, No. 10, 15.05.2017, p. 2730-2742.

Research output: Contribution to journalArticle

Fernandez Diaz-Rullo, Fernando ; Zamberlan, Francesco ; Mewis, Ryan E. ; Fekete, Marianna ; Broche, Lionel ; Cheyne, Lesley A. ; Dall'Angelo, Sergio ; Duckett, Simon B. ; Dawson, Dana ; Zanda, Matteo. / Synthesis and hyperpolarisation of eNOS substrates for quantification of NO production by (1)H NMR spectroscopy. In: Bioorganic & Medicinal Chemistry. 2017 ; Vol. 25, No. 10. pp. 2730-2742.
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abstract = "Hyperpolarization enhances the intensity of the NMR signals of a molecule, whose in vivo metabolic fate can be monitored by MRI with higher sensitivity. SABRE is a hyperpolarization technique that could potentially be used to image nitric oxide (NO) production in vivo. This would be very important, because NO dysregulation is involved in several pathologies, including cardiovascular ones. The nitric oxide synthase (NOS) pathway leads to NO production via conversion of L-arginine into L-citrulline. NO is a free radical gas with a short half-life in vivo (≈5 s), therefore direct NO quantification is challenging. An indirect method – based on quantifying conversion of an L-Arg- to L-Cit-derivative by 1H NMR spectroscopy – is herein proposed. A small library of pyridyl containing L-Arg derivatives was designed and synthesised. In vitro tests showed that compounds 4a–j and 11a–c were better or equivalent substrates for the eNOS enzyme (NO2 − production = 19–46 μM) than native L-Arg (NO2 − production = 25 μM). Enzymatic conversion of L-Arg to L-Cit derivatives could be monitored by 1H NMR. The maximum hyperpolarization achieved by SABRE reached 870-fold NMR signal enhancement, which opens up exciting future perspectives of using these molecules as hyperpolarized MRI tracers in vivo.",
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note = "University of Aberdeen’s Development Trust is gratefully acknowledged for a PhD studentship to F.F.D.-R. We thank Dr Serena Montanari for conducting some preliminary experiments. S.B.D. would like to thank the Wellcome Trust (grant and 098335) for funding.",
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AU - Fernandez Diaz-Rullo, Fernando

AU - Zamberlan, Francesco

AU - Mewis, Ryan E.

AU - Fekete, Marianna

AU - Broche, Lionel

AU - Cheyne, Lesley A.

AU - Dall'Angelo, Sergio

AU - Duckett, Simon B.

AU - Dawson, Dana

AU - Zanda, Matteo

N1 - University of Aberdeen’s Development Trust is gratefully acknowledged for a PhD studentship to F.F.D.-R. We thank Dr Serena Montanari for conducting some preliminary experiments. S.B.D. would like to thank the Wellcome Trust (grant and 098335) for funding.

PY - 2017/5/15

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N2 - Hyperpolarization enhances the intensity of the NMR signals of a molecule, whose in vivo metabolic fate can be monitored by MRI with higher sensitivity. SABRE is a hyperpolarization technique that could potentially be used to image nitric oxide (NO) production in vivo. This would be very important, because NO dysregulation is involved in several pathologies, including cardiovascular ones. The nitric oxide synthase (NOS) pathway leads to NO production via conversion of L-arginine into L-citrulline. NO is a free radical gas with a short half-life in vivo (≈5 s), therefore direct NO quantification is challenging. An indirect method – based on quantifying conversion of an L-Arg- to L-Cit-derivative by 1H NMR spectroscopy – is herein proposed. A small library of pyridyl containing L-Arg derivatives was designed and synthesised. In vitro tests showed that compounds 4a–j and 11a–c were better or equivalent substrates for the eNOS enzyme (NO2 − production = 19–46 μM) than native L-Arg (NO2 − production = 25 μM). Enzymatic conversion of L-Arg to L-Cit derivatives could be monitored by 1H NMR. The maximum hyperpolarization achieved by SABRE reached 870-fold NMR signal enhancement, which opens up exciting future perspectives of using these molecules as hyperpolarized MRI tracers in vivo.

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