A C-terminal cysteine residue is required for peptide-based inhibition of the NGF/TrkA interaction at nM concentrations: implications for peptide-based analgesics

Andrew Poole, Laura Frigotto, Matthew E. Smith, Claudia Baar, Gabriela Ivanova-Berndt, Agnes Jaulent, Catherine Stace, Christopher G. Ullman, Anna V. Hine

Research output: Contribution to journalArticle

Abstract

Inhibition of the NGF/TrkA interaction presents an interesting alternative to the use of non-steroidal anti-inflammatories and/or opioids for the control of inflammatory, chronic and neuropathic pain. Most prominent of the current approaches to this therapy is the antibody Tanezumab, which is a late-stage development humanized monoclonal antibody that targets NGF. We sought to determine whether peptides might similarly inhibit the NGF/TrkA interaction and so serve as future therapeutic leads. Starting from two peptides that inhibit the NGF/TrkA interaction, we sought to eliminate a cysteine residue close to the C-terminal of both sequences, by an approach of mutagenic analysis and saturation mutagenesis of mutable residues. Elimination of cysteine from a therapeutic lead is desirable to circumvent manufacturing difficulties resulting from oxidation. Our analyses determined that the cysteine residue is not required for NGF binding, but is essential for inhibition of the NGF/TrkA interaction at pharmacologically relevant peptide concentrations. We conclude that a cysteine residue is required within potential peptide-based therapeutic leads and hypothesise that these peptides likely act as dimers, mirroring the dimeric structure of the TrkA receptor.
LanguageEnglish
Article number930
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 30 Jan 2019

Fingerprint

Nerve Growth Factor
Cysteine
Analgesics
Peptides
trkA Receptor
Antibodies, Monoclonal, Humanized
Neuralgia
Therapeutics
Mutagenesis
Chronic Pain
Opioid Analgesics
Anti-Inflammatory Agents
Antibodies

Bibliographical note

This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Funding: This work was funded by BBSRC Grant BB/I016481/1 to A.V.H.

Cite this

Poole, Andrew ; Frigotto, Laura ; Smith, Matthew E. ; Baar, Claudia ; Ivanova-Berndt, Gabriela ; Jaulent, Agnes ; Stace, Catherine ; Ullman, Christopher G. ; Hine, Anna V. / A C-terminal cysteine residue is required for peptide-based inhibition of the NGF/TrkA interaction at nM concentrations : implications for peptide-based analgesics. In: Scientific Reports. 2019 ; Vol. 9, No. 1.
@article{1cbeba4082c34bf1b4a3946e2aced47a,
title = "A C-terminal cysteine residue is required for peptide-based inhibition of the NGF/TrkA interaction at nM concentrations: implications for peptide-based analgesics",
abstract = "Inhibition of the NGF/TrkA interaction presents an interesting alternative to the use of non-steroidal anti-inflammatories and/or opioids for the control of inflammatory, chronic and neuropathic pain. Most prominent of the current approaches to this therapy is the antibody Tanezumab, which is a late-stage development humanized monoclonal antibody that targets NGF. We sought to determine whether peptides might similarly inhibit the NGF/TrkA interaction and so serve as future therapeutic leads. Starting from two peptides that inhibit the NGF/TrkA interaction, we sought to eliminate a cysteine residue close to the C-terminal of both sequences, by an approach of mutagenic analysis and saturation mutagenesis of mutable residues. Elimination of cysteine from a therapeutic lead is desirable to circumvent manufacturing difficulties resulting from oxidation. Our analyses determined that the cysteine residue is not required for NGF binding, but is essential for inhibition of the NGF/TrkA interaction at pharmacologically relevant peptide concentrations. We conclude that a cysteine residue is required within potential peptide-based therapeutic leads and hypothesise that these peptides likely act as dimers, mirroring the dimeric structure of the TrkA receptor.",
author = "Andrew Poole and Laura Frigotto and Smith, {Matthew E.} and Claudia Baar and Gabriela Ivanova-Berndt and Agnes Jaulent and Catherine Stace and Ullman, {Christopher G.} and Hine, {Anna V.}",
note = "This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Funding: This work was funded by BBSRC Grant BB/I016481/1 to A.V.H.",
year = "2019",
month = "1",
day = "30",
doi = "10.1038/s41598-018-37585-5",
language = "English",
volume = "9",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",

}

A C-terminal cysteine residue is required for peptide-based inhibition of the NGF/TrkA interaction at nM concentrations : implications for peptide-based analgesics. / Poole, Andrew; Frigotto, Laura; Smith, Matthew E.; Baar, Claudia; Ivanova-Berndt, Gabriela; Jaulent, Agnes; Stace, Catherine; Ullman, Christopher G.; Hine, Anna V.

In: Scientific Reports, Vol. 9, No. 1, 930, 30.01.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A C-terminal cysteine residue is required for peptide-based inhibition of the NGF/TrkA interaction at nM concentrations

T2 - Scientific Reports

AU - Poole, Andrew

AU - Frigotto, Laura

AU - Smith, Matthew E.

AU - Baar, Claudia

AU - Ivanova-Berndt, Gabriela

AU - Jaulent, Agnes

AU - Stace, Catherine

AU - Ullman, Christopher G.

AU - Hine, Anna V.

N1 - This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Funding: This work was funded by BBSRC Grant BB/I016481/1 to A.V.H.

PY - 2019/1/30

Y1 - 2019/1/30

N2 - Inhibition of the NGF/TrkA interaction presents an interesting alternative to the use of non-steroidal anti-inflammatories and/or opioids for the control of inflammatory, chronic and neuropathic pain. Most prominent of the current approaches to this therapy is the antibody Tanezumab, which is a late-stage development humanized monoclonal antibody that targets NGF. We sought to determine whether peptides might similarly inhibit the NGF/TrkA interaction and so serve as future therapeutic leads. Starting from two peptides that inhibit the NGF/TrkA interaction, we sought to eliminate a cysteine residue close to the C-terminal of both sequences, by an approach of mutagenic analysis and saturation mutagenesis of mutable residues. Elimination of cysteine from a therapeutic lead is desirable to circumvent manufacturing difficulties resulting from oxidation. Our analyses determined that the cysteine residue is not required for NGF binding, but is essential for inhibition of the NGF/TrkA interaction at pharmacologically relevant peptide concentrations. We conclude that a cysteine residue is required within potential peptide-based therapeutic leads and hypothesise that these peptides likely act as dimers, mirroring the dimeric structure of the TrkA receptor.

AB - Inhibition of the NGF/TrkA interaction presents an interesting alternative to the use of non-steroidal anti-inflammatories and/or opioids for the control of inflammatory, chronic and neuropathic pain. Most prominent of the current approaches to this therapy is the antibody Tanezumab, which is a late-stage development humanized monoclonal antibody that targets NGF. We sought to determine whether peptides might similarly inhibit the NGF/TrkA interaction and so serve as future therapeutic leads. Starting from two peptides that inhibit the NGF/TrkA interaction, we sought to eliminate a cysteine residue close to the C-terminal of both sequences, by an approach of mutagenic analysis and saturation mutagenesis of mutable residues. Elimination of cysteine from a therapeutic lead is desirable to circumvent manufacturing difficulties resulting from oxidation. Our analyses determined that the cysteine residue is not required for NGF binding, but is essential for inhibition of the NGF/TrkA interaction at pharmacologically relevant peptide concentrations. We conclude that a cysteine residue is required within potential peptide-based therapeutic leads and hypothesise that these peptides likely act as dimers, mirroring the dimeric structure of the TrkA receptor.

UR - https://doi.org/10.17036/researchdata.aston.ac.uk.00000375

UR - http://www.scopus.com/inward/record.url?scp=85060907390&partnerID=8YFLogxK

U2 - 10.1038/s41598-018-37585-5

DO - 10.1038/s41598-018-37585-5

M3 - Article

VL - 9

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

IS - 1

M1 - 930

ER -