Cyclic peptides incorporating phosphotyrosine mimetics as potent and specific inhibitors of the Grb7 breast cancer target

Gabrielle M Watson, Menachem J Gunzburg, Nigus D Ambaye, William S Lucas, Daouda A K Traore, Ketav P Kulkarni, Katie M Cergol, Richard J Payne, Santosh Panjikar, Stephanie C Pero, Patrick Perlmutter, Matthew C Wilce, Jacqueline A Wilce

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The Grb7 adaptor protein is a therapeutic target for both TNBC and HER2+ breast cancer. A nonphosphorylated cyclic peptide 1 (known as G7-18NATE) inhibits Grb7 via targeting the Grb7-SH2 domain, but requires the presence of phosphate ions for both affinity and specificity. Here we report the discovery of malonate bound in the phosphotyrosine binding pocket of the apo-Grb7-SH2 structure. Based on this, we carried out the rational design and synthesis of two analogues of peptide 1 that incorporate carboxymethylphenylalanine (cmF) and carboxyphenylalanine (cF) as mimics of phosphotyrosine (pY). Binding studies using SPR confirmed that affinity for Grb7-SH2 domain is improved up to 9-fold over peptide 1 under physiological phosphate conditions (KD = 2.1-5.7 muM) and that binding is specific for Grb7-SH2 over closely related domains (low or no detectable binding to Grb2-SH2 and Grb10-SH2). X-ray crystallographic structural analysis of the analogue bearing a cmF moiety in complex with Grb7-SH2 has identified the precise contacts conferred by the pY mimic that underpin this improved molecular interaction. Together this study identifies and characterizes the tightest specific inhibitor of Grb7 to date, representing a significant development toward a new Grb7-targeted therapeutic.
Original languageEnglish
Pages (from-to)7707 - 7718
Number of pages12
JournalJournal of Medicinal Chemistry
Volume58
Issue number19
DOIs
Publication statusPublished - 2015

Cite this

Watson, Gabrielle M ; Gunzburg, Menachem J ; Ambaye, Nigus D ; Lucas, William S ; Traore, Daouda A K ; Kulkarni, Ketav P ; Cergol, Katie M ; Payne, Richard J ; Panjikar, Santosh ; Pero, Stephanie C ; Perlmutter, Patrick ; Wilce, Matthew C ; Wilce, Jacqueline A. / Cyclic peptides incorporating phosphotyrosine mimetics as potent and specific inhibitors of the Grb7 breast cancer target. In: Journal of Medicinal Chemistry. 2015 ; Vol. 58, No. 19. pp. 7707 - 7718.
@article{4d05b02e916f4004be0c52a83f3196e9,
title = "Cyclic peptides incorporating phosphotyrosine mimetics as potent and specific inhibitors of the Grb7 breast cancer target",
abstract = "The Grb7 adaptor protein is a therapeutic target for both TNBC and HER2+ breast cancer. A nonphosphorylated cyclic peptide 1 (known as G7-18NATE) inhibits Grb7 via targeting the Grb7-SH2 domain, but requires the presence of phosphate ions for both affinity and specificity. Here we report the discovery of malonate bound in the phosphotyrosine binding pocket of the apo-Grb7-SH2 structure. Based on this, we carried out the rational design and synthesis of two analogues of peptide 1 that incorporate carboxymethylphenylalanine (cmF) and carboxyphenylalanine (cF) as mimics of phosphotyrosine (pY). Binding studies using SPR confirmed that affinity for Grb7-SH2 domain is improved up to 9-fold over peptide 1 under physiological phosphate conditions (KD = 2.1-5.7 muM) and that binding is specific for Grb7-SH2 over closely related domains (low or no detectable binding to Grb2-SH2 and Grb10-SH2). X-ray crystallographic structural analysis of the analogue bearing a cmF moiety in complex with Grb7-SH2 has identified the precise contacts conferred by the pY mimic that underpin this improved molecular interaction. Together this study identifies and characterizes the tightest specific inhibitor of Grb7 to date, representing a significant development toward a new Grb7-targeted therapeutic.",
author = "Watson, {Gabrielle M} and Gunzburg, {Menachem J} and Ambaye, {Nigus D} and Lucas, {William S} and Traore, {Daouda A K} and Kulkarni, {Ketav P} and Cergol, {Katie M} and Payne, {Richard J} and Santosh Panjikar and Pero, {Stephanie C} and Patrick Perlmutter and Wilce, {Matthew C} and Wilce, {Jacqueline A}",
year = "2015",
doi = "10.1021/acs.jmedchem.5b00609",
language = "English",
volume = "58",
pages = "7707 -- 7718",
journal = "Journal of Medicinal Chemistry",
issn = "0022-2623",
publisher = "AMER CHEMICAL SOC",
number = "19",

}

Cyclic peptides incorporating phosphotyrosine mimetics as potent and specific inhibitors of the Grb7 breast cancer target. / Watson, Gabrielle M; Gunzburg, Menachem J; Ambaye, Nigus D; Lucas, William S; Traore, Daouda A K; Kulkarni, Ketav P; Cergol, Katie M; Payne, Richard J; Panjikar, Santosh; Pero, Stephanie C; Perlmutter, Patrick; Wilce, Matthew C; Wilce, Jacqueline A.

In: Journal of Medicinal Chemistry, Vol. 58, No. 19, 2015, p. 7707 - 7718.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Cyclic peptides incorporating phosphotyrosine mimetics as potent and specific inhibitors of the Grb7 breast cancer target

AU - Watson, Gabrielle M

AU - Gunzburg, Menachem J

AU - Ambaye, Nigus D

AU - Lucas, William S

AU - Traore, Daouda A K

AU - Kulkarni, Ketav P

AU - Cergol, Katie M

AU - Payne, Richard J

AU - Panjikar, Santosh

AU - Pero, Stephanie C

AU - Perlmutter, Patrick

AU - Wilce, Matthew C

AU - Wilce, Jacqueline A

PY - 2015

Y1 - 2015

N2 - The Grb7 adaptor protein is a therapeutic target for both TNBC and HER2+ breast cancer. A nonphosphorylated cyclic peptide 1 (known as G7-18NATE) inhibits Grb7 via targeting the Grb7-SH2 domain, but requires the presence of phosphate ions for both affinity and specificity. Here we report the discovery of malonate bound in the phosphotyrosine binding pocket of the apo-Grb7-SH2 structure. Based on this, we carried out the rational design and synthesis of two analogues of peptide 1 that incorporate carboxymethylphenylalanine (cmF) and carboxyphenylalanine (cF) as mimics of phosphotyrosine (pY). Binding studies using SPR confirmed that affinity for Grb7-SH2 domain is improved up to 9-fold over peptide 1 under physiological phosphate conditions (KD = 2.1-5.7 muM) and that binding is specific for Grb7-SH2 over closely related domains (low or no detectable binding to Grb2-SH2 and Grb10-SH2). X-ray crystallographic structural analysis of the analogue bearing a cmF moiety in complex with Grb7-SH2 has identified the precise contacts conferred by the pY mimic that underpin this improved molecular interaction. Together this study identifies and characterizes the tightest specific inhibitor of Grb7 to date, representing a significant development toward a new Grb7-targeted therapeutic.

AB - The Grb7 adaptor protein is a therapeutic target for both TNBC and HER2+ breast cancer. A nonphosphorylated cyclic peptide 1 (known as G7-18NATE) inhibits Grb7 via targeting the Grb7-SH2 domain, but requires the presence of phosphate ions for both affinity and specificity. Here we report the discovery of malonate bound in the phosphotyrosine binding pocket of the apo-Grb7-SH2 structure. Based on this, we carried out the rational design and synthesis of two analogues of peptide 1 that incorporate carboxymethylphenylalanine (cmF) and carboxyphenylalanine (cF) as mimics of phosphotyrosine (pY). Binding studies using SPR confirmed that affinity for Grb7-SH2 domain is improved up to 9-fold over peptide 1 under physiological phosphate conditions (KD = 2.1-5.7 muM) and that binding is specific for Grb7-SH2 over closely related domains (low or no detectable binding to Grb2-SH2 and Grb10-SH2). X-ray crystallographic structural analysis of the analogue bearing a cmF moiety in complex with Grb7-SH2 has identified the precise contacts conferred by the pY mimic that underpin this improved molecular interaction. Together this study identifies and characterizes the tightest specific inhibitor of Grb7 to date, representing a significant development toward a new Grb7-targeted therapeutic.

UR - http://www.ncbi.nlm.nih.gov/pubmed/26359549

U2 - 10.1021/acs.jmedchem.5b00609

DO - 10.1021/acs.jmedchem.5b00609

M3 - Article

VL - 58

SP - 7707

EP - 7718

JO - Journal of Medicinal Chemistry

JF - Journal of Medicinal Chemistry

SN - 0022-2623

IS - 19

ER -