Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014 Jan 28;12(4):566-73.
doi: 10.1039/c3ob42168f.

Convergent diversity-oriented side-chain macrocyclization scan for unprotected polypeptides

Yekui Zou  1 , Alexander M Spokoyny, Chi Zhang, Mark D Simon, Hongtao Yu, Yu-Shan Lin, Bradley L Pentelute
Affiliations

Convergent diversity-oriented side-chain macrocyclization scan for unprotected polypeptides

Yekui Zou et al. Org Biomol Chem. .

Abstract

Here we describe a general synthetic platform for side-chain macrocyclization of an unprotected peptide library based on the SNAr reaction between cysteine thiolates and a new generation of highly reactive perfluoroaromatic small molecule linkers. This strategy enabled us to simultaneously "scan" two cysteine residues positioned from i, i + 1 to i, i + 14 sites in a polypeptide, producing 98 macrocyclic products from reactions of 14 peptides with 7 linkers. A complementary reverse strategy was developed; cysteine residues within the polypeptide were first modified with non-bridging perfluoroaryl moieties and then commercially available dithiol linkers were used for macrocyclization. The highly convergent, site-independent, and modular nature of these two strategies coupled with the unique chemoselectivity of a SNAr transformation allows for the rapid diversity-oriented synthesis of hybrid macrocyclic peptide libraries with varied chemical and structural complexities.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Convergent diversity-oriented synthetic (DOS) platform for a peptide macrocyclization scan utilizing a library of chemically tailorable bifunctional linkers. AAx refers to a specific amino acid residue in the peptide chain sequence.
Figure 2
Figure 2
An unprotected peptide macrocyclization scan enabled by SNAr transformation between thiols and activated perfluoroaromatics: (a) the reaction scheme highlighting the regioselectivity of the SNAr process leading to preferential substitution at para position of the pentafluoroaryl moiety with respect to the R substituent; (b) the reactivity trend of perfluoroaryl-based electrophiles governed by two independent activation modes; (c) the library of bifunctional perfluoroaryl-based linkers containing thioether moieties capable of activating corresponding para-CF moiety towards nucleophilic attack via resonance stabilization of the Meisenheimer intermediate; (d) two independent strategies designed to probe macrocyclization scan with linkers La-Lg; and (e) 14 model cysteine-containing unprotected peptides used for the studies.
Figure 3
Figure 3
LC-MS chromatograms (total ion current) of purified peptide 7 and corresponding macrocyclization reactions with linkers La-Lg analyzed in situ. Peaks labeled as * represent oxidized disulfide by-product.
Figure 4
Figure 4
LC-MS chromatograms (total ion current) of purified peptide 14 and corresponding macrocyclization reactions with linkers La-Lg analyzed in situ. Peaks labeled as * represent oxidized disulfide by-product.
Figure 5
Figure 5
Bar graph summary of the macrocyclization scan with peptides 1–14 and linkers La-Lg. Number within each bar represents corresponding yield of the macrocyclization product determined by LC-MS analysis of the unpurified product mixture. Note, for yields denoted with *, re-optimized conditions were employed (see SI).
Figure 6
Figure 6
Synthesis of cysteine perfluorinated peptides 7a′ and 7b′, LC-MS chromatograms (total ion current) of purified peptides 7a′ and 7b′, and corresponding macrocyclization reactions with dithiol linkers (highlighted in grey) analyzed in situ.
See this image and copyright information in PMC

References

    1. Baeriswyl V, Heinis C. Chem Med Chem. 2013;8:377–384. - PubMed
    2. Smith JM, Frost JR, Fasan R. J Org Chem. 2013;78:3525–3531. - PubMed
    1. Craik DJ. Science. 2006;311:1563–1564. - PubMed

    1. Selected reviews: Lambert NJ, Mitchell JP, Roberts KD. J Chem Soc, Perkin Trans. 2001;1:471–484.Davies JS. J Pept Sci. 2003;9:471–501.White CJ, Yudin AK. Nature Chem. 2011;3:509–524.

    1. Representative examples: Timmerman P, Beld J, Pujik WC, Meloen RH. Chem Bio Chem. 2005;6:821–824.Muppidi A, Doi K, Edwardraja S, Drake EJ, Gulick AM, Wang HG, Lin Q. J Am Chem Soc. 2012;134:14734–14737.Heinis C, Rutherford T, Freund S, Winter G. Nature Chem Bio. 2009;5:502–507.Smeenk LEJ, Dailly N, Hiemstra H, van Maarseveen JH, Timmerman P. Org Lett. 2012;14:1194–1197.Jo H, Meinhardt N, Wu Y, Kulkarni S, Hu X, Low KE, Davies PL, DeGrado WF, Greenbaum DC. J Am Chem Soc. 2012;134:17704–17713.Kawakami T, Ishizawa T, Fujino T, Reid PC, Suga H, Murakami H. ACS Chem Bio. 2013;8:1205–1214.

    1. Representative examples, olefin metathesis: Miller SJ, Blackwell HE, Grubbs RH. J Am Chem Soc. 1996;118:9606–9614.Kim YW, Grossmann TN, Verdine GL. Nature Prot. 2011;6:761–771.Kim YW, Kutchukian PS, Verdine GL. Org Lett. 2010;12:3046–3049.Kim YW, Verdine GL. Bioorg & Med Chem Lett. 2009;19:2533–2536.Bautista AD, Appelbaum JS, Craig CJ, Michel J, Schepartz A. J Am Chem Soc. 2010;132:2904–2906.Moellering RE, Cornejo M, Davis TN, Del Bianco C, Aster JC, Blacklow SC, Kung AL, Gilliland DG, Verdine GL, Bradner JE. Nature. 2009;462:182–188.Reichwein JF, Wels B, Kruijtzer JAW, Versluis C, Liskamp RMJ. Angew Chem, Int Ed. 1999;38:3684–3687.Yeo DJ, Warriner SL, Wilson AJ. Chem Commun. 2013;49:9131–9133.aryl cross-coupling: Dong H, Limberakis C, Price D, James K. Chem Commun. 2012;48:11644–11646.Meyer FM, Collins JC, Borin B, Bradow J, Liras S, Limberakis C, Mathiowetz AM, Philippe L, Price D, Song K, James K. J Org Chem. 2012;77:3099–3114.

Publication types