Targeting a hydrophobic patch on the surface of the Grb2-SH2 domain leads to high-affinity phosphotyrosine-containing peptide ligands

Fretz, Heinz; Furet, Pascal; Schoepfer, Joseph; García‐Echeverría, Carlos; Rahuel, Joseph; Caravatti, Giorgio

doi:10.1007/0-306-46862-x_309

Cited by 2 publications

(7 citation statements)

References 2 publications

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“…39,40 The linear Shc(Y317) peptide D-D-P-S-pY-V-N-V-Q (24) was employed as a reference and gave an IC 50 of 1.3 µM in this system. Consistent with previous reports, 19,20 the parent pTyr-containing 18b also exhibited very potent binding affinity (IC 50 ) 0.07 µM). The importance of the "phosphate pharmacophore" to the overall binding of 18b was confirmed by the loss of all measurable binding affinity upon removal of the phosphate group (compound 18a).…”

Section: Resultssupporting

confidence: 92%

“…A series of new inhibitors 18c − f , 20d , and 20f incorporating pTyr mimetics 15c − f was prepared, and Grb2 SH2 domain binding affinities were determined by surface plasmon reasonance (Figures and ). , The linear Shc(Y317) peptide D-D-P-S-pY-V-N-V-Q ( 24 ) was employed as a reference and gave an IC 50 of 1.3 μM in this system. Consistent with previous reports, , the parent pTyr-containing 18b also exhibited very potent binding affinity (IC 50 = 0.07 μM). The importance of the “phosphate pharmacophore” to the overall binding of 18b was confirmed by the loss of all measurable binding affinity upon removal of the phosphate group (compound 18a ).…”

Section: Resultssupporting

confidence: 91%

“…A Novartis group has accordingly reported several high-affinity pTyr-containing ligands, which, in addition to having in common a pTyr residue that binds in the pTyr pocket and an Asn residue in the pTyr + 2 position, contain other features that enhance Grb2 binding affinity. 36 Among these features are elements which induce β-bend conformations, as shown by cyclic peptide 21 (IC 50 ) 0.37 µM) 37 and β-bend-mimicking open-chain tripeptides typified by 22 (IC 50 ) 0.011 µM) 19,20 which utilize a 1-aminocyclohexanecarboxylic acid in the pTyr + 1 position to facilitate β-bend formation. 38 This latter type of structure provided an ideal starting point for the current study, which is focused on interactions within the pTyr binding pocket.…”

Section: Resultsmentioning

confidence: 99%

“…Plasmon Reasonace Binding Studies. To examine pTyr mimetics in the context of Grb2 SH2 inhibitors, the recently disclosed high-affinity pTyr-containing tripeptide 18b 19,20 was chosen as a model platform on which to append phosphotyrosyl-mimicking analogues. A series of new inhibitors 18c-f, 20d, and 20f incorporating pTyr mimetics 15c-f was prepared, and Grb2 SH2 domain binding affinities were determined by surface plasmon reasonance (Figures 1 and 2).…”

Section: Resultsmentioning

confidence: 99%

“…pTyr-containing β-bend mimetic 18b is within the structural family of Grb2 SH2 domain antagonists previously synthesized by solid-phase techniques. 19,20 Despite repeated attempts, we were unable to achieve the synthesis of 18b by solid-phase methods while maintaining the N-terminal naphthylpropylamide. This failure was most probably related to steric crowding induced by the naphthylpropylamido group proximal to the R-amino and carboxyl moieties of the β-bend-forming 1-aminocyclohexanecarboxylic acid residue (13).…”

Section: Synthesismentioning

confidence: 99%

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Potent Inhibition of Grb2 SH2 Domain Binding by Non-Phosphate-Containing Ligands

et al. 1998

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Development of Grb2 Src homology 2 (SH2) domain binding inhibitors has important implications for treatment of a variety of diseases, including several cancers. In cellular studies, inhibitors of Grb2 SH2 domain binding have to date been large, highly charged peptides which relied on special transport devices for cell membrane penetration. Work presented in the current study examines a variety of pTyr mimetics in the context of a high-affinity Grb2 binding platform. Among the analogues studied are new non-phosphorus-containing pTyr mimetics 23a and 23b which, when incorporated into tripeptide structures 18f and 20f, are able to inhibit Grb2 SH2 domain binding with affinities among the best yet reported for non-phosphorus-containing SH2 domain inhibitors (IC50 values of 6.7 and 1.3 microM, respectively). The present study has also demonstrated the usefulness of the Nalpha-oxalyl group as an auxiliary which enhances the binding potency of both phosphorus- and non-phosphorus-containing pTyr mimetics. When combined with the (phosphonomethyl)phenylalanine (Pmp) residue to give analogues such as L-20d, potent inhibition of Grb2 SH2 domain binding can be achieved both in extracellular assays using isolated Grb2 SH2 domain protein and in intracellular systems measuring the association of endogenous Grb2 with its cognate p185erbB-2 ligand. These latter effects can be achieved at micromolar to submicromolar concentrations without prodrug derivatization. The oxalyl-containing pTyr mimetics presented in this study should be of general usefulness for the development of other Grb2 SH2 domain antagonists, independent of the beta-bend-mimicking platform utilized for their display.

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Section: Resultssupporting

confidence: 92%

Section: Resultssupporting

confidence: 91%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Synthesismentioning

confidence: 99%

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Potent Inhibition of Grb2 SH2 Domain Binding by Non-Phosphate-Containing Ligands

et al. 1998

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Utilization of a β-Aminophosphotyrosyl Mimetic in the Design and Synthesis of Macrocyclic Grb2 SH2 Domain-Binding Peptides

et al. 2003

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Grb2 SH2 domains are protein-docking modules that exert important functions in both normal and pathogenic signal transduction processes. Development of synthetic Grb2 SH2 domain binding ligands is being pursued by several groups as potential new therapies for a variety of diseases, including certain cancers. In these efforts, macrocyclization has been successfully utilized to take advantage of preferential recognition by Grb2 SH2 domains of ligands in beta-bend conformations. Recent examples of this approach include olefin-metathesis-derived macrocycles that employ ring closure at the beta-position of key pTyr-mimicking residues. In the current study, a novel phosphatase-stable beta-amino-pTyr mimetic designated "Pmp(beta)" was utilized to prepare variants of previously reported olefin-metathesis-derived macrocycles. An initial set of simplified cyclic peptides lacking key naphthyl side chain functionality was first synthesized to determine optimum ring size, with results indicating that a four-unit ring-closing segment was appropriate. On the basis of these findings, macrolactamization was undertaken with a more highly functionalized, naphthyl-containing gamma-amino acid analogue. The resulting cyclic beta-amino peptide is the first of a new class of pTyr-mimetic-containing ligands that may have utility in the development of antagonists of both Grb2 SH2 domains and other pTyr-dependent signaling systems.

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Targeting a hydrophobic patch on the surface of the Grb2-SH2 domain leads to high-affinity phosphotyrosine-containing peptide ligands

Cited by 2 publications

References 2 publications

Potent Inhibition of Grb2 SH2 Domain Binding by Non-Phosphate-Containing Ligands

Potent Inhibition of Grb2 SH2 Domain Binding by Non-Phosphate-Containing Ligands

Utilization of a β-Aminophosphotyrosyl Mimetic in the Design and Synthesis of Macrocyclic Grb2 SH2 Domain-Binding Peptides

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