Phage Selection of Peptide Macrocycles against β‐Catenin To Interfere with Wnt Signaling

Bertoldo, Davide; Khan, Maola M. G.; Dessen, Pierre; Held, Werner; Huelsken, Joerg; Heinis, Christian

doi:10.1002/cmdc.201500557

Cited by 34 publications

(27 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The remainder of the screens were varied in their models and approach, encompassing virtual screen techniques to identify potential modifiers of the DKK1 interactions to the use of reporters in zebrafish scales [ 15 , 16 ]. In one case, the use of phage display allowed for the screening of 12 billion bicyclic peptides that might interact with the β-catenin ARM repeats [ 17 ].…”

Section: Resultsmentioning

confidence: 99%

Wnt Drug Discovery: Weaving Through the Screens, Patents and Clinical Trials

Green

Farr

et al. 2016

Cancers

View full text Add to dashboard Cite

The Wnt signaling pathway is intricately involved in many aspects of development and is the root cause of an increasing number of diseases. For example, colorectal cancer is the second leading cause of death in the industrialized world and aberration of Wnt signaling within the colonic stem cell is the cause of more than 90% of these cancers. Despite our advances in successfully targeting other pathways, such as Human Epidermal Growth Factor Receptor 2 (HER2), there are no clinically relevant therapies available for Wnt-related diseases. Here, we investigated where research activities are focused with respect to Wnt signaling modulators by searching the United States Patent and Trade Office (USPTO) for patents and patent applications related to Wnt modulators and compared this to clinical trials focusing on Wnt modulation. We found that while the transition of intellectual property surrounding the Wnt ligand-receptor interface to clinical trials is robust, this is not true for specific inhibitors of β-catenin, which is constitutively active in many cancers. Considering the ubiquitous use of the synthetic T-cell Factor/Lymphoid Enhancer Factor (TCF/Lef) reporter system and its success in identifying novel modulators in vitro, we speculate that this model of drug discovery does not capture the complexity of in vivo Wnt signaling that may be required if we are to successfully target the Wnt pathway in the clinic. Notwithstanding, increasingly more complex models are being developed, which may not be high throughput, but more pragmatic in our pursuit to control Wnt signaling.

show abstract

Section: Resultsmentioning

confidence: 99%

Wnt Drug Discovery: Weaving Through the Screens, Patents and Clinical Trials

Green

Farr

et al. 2016

Cancers

View full text Add to dashboard Cite

show abstract

“…B. mithilfe von Split-Mix-Bibliotheken bicyclischer Peptide identifiziert. B. S.-Aureus-Sortase A, [70] Her2, [71] b-Catenin, [72] Urokinase, [73] Kallikrein im Plasma [67a, 74] und Notch1, [75] zu finden. [66] Randomisierte Peptidbibliotheken, die drei konservierte Cysteinreste enthalten, kçnnen auch auf filamentçsen Phagen präsentiert und einer Cyclisierung mittels thiolreaktiver organischer Gerüststrukturen unterzogen werden (Abbildung 5a).…”

Section: Bicyclische Gerüststrukturenunclassified

“…Diese Bibliotheken wurden herangezogen, um Bindungspartner füreine Vielzahl biologischer Zielstrukturen, z. B. S.-Aureus-Sortase A, [70] Her2, [71] b-Catenin, [72] Urokinase, [73] Kallikrein im Plasma [67a, 74] und Notch1, [75] zu finden.…”

Section: Bicyclische Gerüststrukturenunclassified

Neue Modalitäten für schwierige Zielstrukturen in der Wirkstoffentwicklung

et al. 2017

View full text Add to dashboard Cite

Unsere sich stetig erweiternden Kenntnisse über biologische Systeme haben eine Vielzahl an hoch interessanten neuen biologischen Zielstrukturen aufgedeckt, deren Modulation möglicherweise neuartige Therapiemöglichkeiten für viele Krankheiten erschließt. Diese Angriffspunkte umfassen insbesondere Protein‐Protein‐ und Protein‐Nukleinsäure‐Wechselwirkungen, die jedoch durch klassische niedermolekulare Substanzen oftmals nicht adressierbar sind. Andere Substanzklassen oder Modalitäten werden daher benötigt, um diese Ziele anzuvisieren. Einige akademische Forschungsgruppen und pharmazeutische Unternehmen greifen daher zunehmend das Konzept der so genannten “neuen Modalitäten” auf. Dieser Aufsatz definiert erstmals diesen Begriff, der neuartige Peptidgerüststrukturen, Oligonukleotide, Hybride, molekulare Konjugate sowie auf neuartige Weise eingesetzte, klassische niedermolekulare Verbindungen berücksichtigt. Wir stellen die repräsentativsten Beispiele dieser Modalitäten vor, die auf große Bindungsoberflächen, wie sie in Protein‐Protein‐Wechselwirkungen vorkommen, sowie auf zentrale biologische Zellregulationsvorgänge abzielen.

show abstract

“…14 However, peptide cyclization has been emphasized to a lesser extent, [15][16][17][18][19] and has been predominantly achieved by the formation of disulfide bonds or nucleophilic substitution using bifunctional linkers on lysine and cysteine residues. 3,6,20,215,22,23 More recently, enzymatic cyclization has been used to construct peptide macrocycles on the surface of yeast 24 and bacteria, 25 or within bacteria. 26 In one study, a library of lanthipeptides was cyclized using the ProcM enzyme, resulting in macrocyclic peptides displayed on the surface of yeast cells that were screened to identify ligands for αvβ3 integrin.…”

Section: Introductionmentioning

confidence: 99%

Screening of yeast display libraries of enzymatically-cyclized peptides to discover macrocyclic peptide ligands

Bowen

Schneible

Labar

et al. 2020

Preprint

View full text Add to dashboard Cite

Abstract. We present the construction and screening of yeast display libraries of cyclic peptides wherein site-selective enzymatic cyclization of linear peptides is achieved using bacterial transglutaminase. To this end, we developed two alternative routes, namely (i) yeast display of linear peptides followed by treatment with recombinant transglutaminase in solution; or (ii) intracellular co-expression of linear peptides and transglutaminase to achieve cyclization in the endoplasmic reticulum prior to yeast surface display. The cyclization yield was evaluated via orthogonal detection of epitope tags integrated in the yeast-displayed peptides by flow cytometry, and via comparative cleavage of cyclic vs. linear peptides by tobacco etch virus (TEV) protease. Subsequently, yeast display libraries of transglutaminase-cyclized peptides were screened to isolate binders to the N-terminal region of the Yes-Associated Protein (YAP) and its WW domains using magnetic selection and fluorescence activated cell sorting (FACS). The identified cyclic peptide cyclo[E-LYLAYPAH-K] featured a KD of 1.67 μM for YAP and 0.84 μM for WW as well as high binding selectivity against albumin and lysozyme. These results demonstrate the usefulness of yeast surface display for screening transglutaminase-cyclized peptide libraries, and efficient identification of cyclic peptide ligands.

show abstract

Phage Selection of Peptide Macrocycles against β‐Catenin To Interfere with Wnt Signaling

Cited by 34 publications

References 21 publications

Wnt Drug Discovery: Weaving Through the Screens, Patents and Clinical Trials

Wnt Drug Discovery: Weaving Through the Screens, Patents and Clinical Trials

Neue Modalitäten für schwierige Zielstrukturen in der Wirkstoffentwicklung

Screening of yeast display libraries of enzymatically-cyclized peptides to discover macrocyclic peptide ligands

Contact Info

Product

Resources

About