TY - JOUR
T1 - A Strategy to Select Macrocyclic Peptides Featuring Asymmetric Molecular Scaffolds as Cyclization Units by Phage Display
AU - Oppewal, Titia Rixt
AU - Jansen, Ivar D.
AU - Hekelaar, Johan
AU - Mayer, Clemens
N1 - Funding Information:
C.M. and T.R.O. are thankful to Dr. Johan Kemmink and Pieter van der Meulen for their help in performing and analyzing the NMR experiments. The authors thank Gea Schuurman-Wolters for assistance with ITC measurements and Rudy Rubini for his guidance in molecular biology operations. C.M. is grateful to Prof. Christian Heinis for providing phage vectors fdg3p0ss21 and fd0D1D2 with permission from Prof. Franz. X. Schmid. C.M. and T.R.O. acknowledge Anthony Kromodimedjo and Catalina Bigalke for the synthesis of cyclization unit 3 as well as Mark Folkertsma for his contribution to the protocol for the construction of phage libraries. C.M. acknowledges the Netherlands Organization for Scientific Research for financial support (NWO, Veni grant 722.017.007).
Publisher Copyright:
© 2022 American Chemical Society. All rights reserved.
PY - 2022/3/2
Y1 - 2022/3/2
N2 - Macrocyclic peptides (MPs) have positioned themselves as a privileged class of compounds for the discovery of therapeutics and development of chemical probes. Aided by the development of powerful selection strategies, high-affinity binders against biomolecular targets can readily be elicited from massive, genetically encoded libraries by affinity selection. For example, in phage display, MPs are accessed on the surface of whole bacteriophages via disulfide formation, the use of (symmetric) crosslinkers, or the incorporation of non-canonical amino acids. To facilitate a straightforward cyclization of linear precursors with asymmetric molecular scaffolds, which are often found at the core of naturally occurring MPs, we report an efficient two-step strategy to access MPs via the programmed modification of a unique cysteine residue and an N-terminal amine. We demonstrate that this approach yields MPs featuring asymmetric cyclization units from both synthetic peptides and when linear precursors are appended onto a phage-coat protein. Finally, we showcase that our cyclization strategy is compatible with traditional phage-display protocols and enables the selection of MP binders against a model target protein from naïve libraries. By enabling the incorporation of non-peptidic moieties that (1) can serve as cyclization units, (2) provide interactions for binding, and/or (3) tailor pharmacological properties, our head-to-side-chain cyclization strategy provides access to a currently under-explored chemical space for the development of chemical probes and therapeutics.
AB - Macrocyclic peptides (MPs) have positioned themselves as a privileged class of compounds for the discovery of therapeutics and development of chemical probes. Aided by the development of powerful selection strategies, high-affinity binders against biomolecular targets can readily be elicited from massive, genetically encoded libraries by affinity selection. For example, in phage display, MPs are accessed on the surface of whole bacteriophages via disulfide formation, the use of (symmetric) crosslinkers, or the incorporation of non-canonical amino acids. To facilitate a straightforward cyclization of linear precursors with asymmetric molecular scaffolds, which are often found at the core of naturally occurring MPs, we report an efficient two-step strategy to access MPs via the programmed modification of a unique cysteine residue and an N-terminal amine. We demonstrate that this approach yields MPs featuring asymmetric cyclization units from both synthetic peptides and when linear precursors are appended onto a phage-coat protein. Finally, we showcase that our cyclization strategy is compatible with traditional phage-display protocols and enables the selection of MP binders against a model target protein from naïve libraries. By enabling the incorporation of non-peptidic moieties that (1) can serve as cyclization units, (2) provide interactions for binding, and/or (3) tailor pharmacological properties, our head-to-side-chain cyclization strategy provides access to a currently under-explored chemical space for the development of chemical probes and therapeutics.
UR - http://www.scopus.com/inward/record.url?scp=85125415824&partnerID=8YFLogxK
U2 - 10.1021/jacs.1c12822
DO - 10.1021/jacs.1c12822
M3 - Article
C2 - 35171585
AN - SCOPUS:85125415824
VL - 144
SP - 3644
EP - 3652
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
SN - 0002-7863
IS - 8
ER -