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A Convenient Approach to Prepare Topologically Segregated Bilayer Beads for One-Bead Two-Compound Combinatorial Peptide Libraries

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Abstract

One-bead one-compound (OBOC) combinatorial peptide libraries have been used to identify ligands and modulators for a wide variety of biological targets. While being very efficient with linear peptides, OBOC libraries with N-terminally blocked peptides or with unsequenceable building blocks require encoding. To fully exploit OBOC combinatorial methods with cyclic peptides and peptidomimetics, topologically segregated bilayer beads have been developed. This strategy offers the opportunity to synthesize two compounds per bead, i.e. with one compound exposed on the bead surface for screening, and the other one found within the inner layer as a tag for sequencing and compound identification. Bead segregation often involves the use of unstable derivatives or requires a series of protection–deprotection steps. In order to expedite and optimize bead segregation, the performance of various reagents has been studied. The results obtained herein show that bead segregation can be efficiently performed with commercially available reagents. Finally, in order to control outer/inner layer ratios in segregated beads, the effects of different parameters have been evaluated. We report a straightforward and efficient procedure to prepare topologically segregated bilayer beads in a wide range of controllable, predictable, and reproducible outer versus inner ratios.

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Scheme 1
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Abbreviations

APCI:

Atmospheric pressure chemical ionization

B:

β-Alanine

Boc:

Tert-butyloxycarbonyl

tBu:

Tert-butyl

DBU:

8-Diazabicyclo[5.4.0]undec-7-ene

DCC:

Dicyclohexylcarbodiimide

DCM:

Dichloromethane

DIPEA:

N,N-diisopropylethylamine

DMF:

N,N-dimethylformamide

Et2O:

Diethyl ether

Fmoc:

9-Fluorenylmethyloxycarbonyl

h:

Homoserine lactone

HCTU:

2-(6-Chloro-1H-benzotriazole-1-yl)-1,1,3,3-tetramethylaminium hexafluorophosphate

HOBt:

1-Hydroxybenzotriazole

HOSu:

N-hydroxysuccinimide

MALDI-TOF:

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry

MeCN:

Acetonitrile

MeOH:

Methanol

NMM:

4-Methylmorpholine

OBOC:

One-bead one-compound

OBTC :

One-bead two-compound

PyBOP:

Benzotriazol-1-yl-oxy-tris(pyrrolidino)phosphonium

TFA:

Trifluoroacetic acid

TG:

TentaGel resin

TIS:

Triisopropylsilane

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Acknowledgments

The authors gratefully acknowledge the Banting Research Foundation, the Fonds de recherche en Santé du Québec (FRSQ) and the Fonds Québécois de recherche en Nature et Technologie (FQRNT) for financial support. Anick Girard thanks the Fonds d’enseignement et de recherche de la Faculté de pharmacie de l’Université Laval for postgraduate Scholarships. The authors are grateful to Dr. Richard Poulin for his help in the preparation of this manuscript.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Faculty of Pharmacy, Université Laval, Pavillon Vandry, Quebec City, QC, G1V 0A6, Canada

    François Bédard, Anick Girard & Éric Biron

  2. Laboratory of Medicinal Chemistry, CHUQ Research Center (CHUL Section), 2705 Boulevard Laurier, Quebec City, QC, G1V 4G2, Canada

    François Bédard, Anick Girard & Éric Biron

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  1. François Bédard
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Correspondence to Éric Biron.

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Bédard, F., Girard, A. & Biron, É. A Convenient Approach to Prepare Topologically Segregated Bilayer Beads for One-Bead Two-Compound Combinatorial Peptide Libraries. Int J Pept Res Ther 19, 13–23 (2013). https://doi.org/10.1007/s10989-012-9316-x

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