Evaluation of Lipopeptides as Toll-like Receptor 2 Ligands

Title:Evaluation of Lipopeptides as Toll-like Receptor 2 Ligands

Volume: 14 Issue: 7

Author(s): Waleed M. Hussein, Phil M. Choi, Cheng Zhang, Mei Su, Emma Sierecki, Wayne Johnston, Vincent Fagan, Kirill Alexandrov, Mariusz Skwarczynski, Yann Gambin, Istvan Toth and Pavla Simerska*

Affiliation:

• School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072,Australia

Keywords: AlphaScreen assay, lipoamino acid, lipopeptide, peptide synthesis, TLR2 agonists, vaccines.

Abstract: Background: Peptide-based vaccines are considered to be the next generation of modern immunizations, as they are safe, easy to produce and well-defined. However, due to their weak immunogenic effect, it is important to first develop an appropriate adjuvant for peptide-based vaccines.

Objective: The aim of this work was to synthesize a series of four adjuvanting moieties as alkyne derivatives, incorporating dipalmitoyl serine (DPS), 1,3-diglyceride (DG), two hexadecane lipoamino acids (diLAA), and 2,3-dipalmitoyl-S-glycerylcysteine (Pam2Cys). Next aim was to synthesize and attach the azide derivative of biotinylated J14 peptide (model B-cell epitope) to the alkynes through copper- catalyzed alkyne-azide 1,3-dipolar cycloaddition (CuAAC) reaction. Final aim was to test the ability of the final biotin labeled conjugates to directly interact with in vitro expressed TLR2 and 8 using AlphaScreen proximity assay.

Method: All of the peptides were synthesized by manual stepwise solid phase peptide synthesis (SPPS) on rink amide MBHA resin using HATU/DIPEA Fmoc-chemistry. The target compounds were synthesized in a solution phase using CuAAC reaction.

Results: Pam2Cys analogue bound to TLR2 as expected. Analogues of DPS and C16-LAA showed also affinity to TLR2, while it did not bind to the control protein (TLR8), demonstrating ability of the DPS and C16-LAA to be recognized by TLR2.

Conclusion: Four alkyne derivatives of lipids were successfully synthesized and coupled to a biotinylated J14 peptide to give a series of self-adjuvanting ligands. These ligands showed different affinity to TLR2 upon testing by AlphaScreen assay. The DPS derivative showed the most promising affinity in comparison to the standard TLR2 agonist, Pam2Cys.

Cite this article as:

Hussein M. Waleed, Choi M. Phil , Zhang Cheng, Su Mei, Sierecki Emma , Johnston Wayne, Fagan Vincent, Alexandrov Kirill, Skwarczynski Mariusz , Gambin Yann, Toth Istvan and Simerska Pavla *, Evaluation of Lipopeptides as Toll-like Receptor 2 Ligands, Current Drug Delivery 2017; 14 (7) . https://dx.doi.org/10.2174/1567201813666160804114107