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A biomimetic approach for enhancing the in vivo half-life of peptides

Abstract

The tremendous therapeutic potential of peptides has not yet been realized, mainly owing to their short in vivo half-life. Although conjugation to macromolecules has been a mainstay approach for enhancing protein half-life, the steric hindrance of macromolecules often harms the binding of peptides to target receptors, compromising the in vivo efficacy. Here we report a new strategy for enhancing the in vivo half-life of peptides without compromising their potency. Our approach involves endowing peptides with a small molecule that binds reversibly to the serum protein transthyretin. Although there are a few molecules that bind albumin reversibly, we are unaware of designed small molecules that reversibly bind other serum proteins and are used for half-life extension in vivo. We show here that our strategy was effective in enhancing the half-life of an agonist for GnRH receptor while maintaining its binding affinity, which was translated into superior in vivo efficacy.

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Figure 1: Crystal structure of hTTR bound to AG10, and effect of binding to TTR on the half-life of AG10.
Figure 2: TLHE1 and its peptide conjugates bind selectively to hTTR in buffer and human serum.
Figure 3: Binding to TTR increased the stability of TLHE1–peptides 5, 6 and 7 in vitro, and extended the t1/2 of 7 in rats.
Figure 4: Compound 8 preferentially binds to GnRH-R over hTTR and does not interfere with holo-RBP-TTR interaction.
Figure 5: Compound 8 displayed extended t1/2 and superior efficacy in rats.

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Acknowledgements

This work was supported by a New Investigator Award from the American Association of Colleges of Pharmacy and the US National Institutes of Health grant 1R15GM110677-01 (M.M.A.). The support by a National Science Foundation Instrumentation grant (NSF-MRI-0722654) is gratefully acknowledged.

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Contributions

M.M.A. conceived and supervised the project. M.M.A, S.C.P. and M.R.M. designed the experiments. S.C.P., M.R.M. and A.P. performed the chemical synthesis, PK studies and biological assays. H.J. and J.T. performed the modeling study. J.D. and P.B. helped with mass spectrometry analysis. N.R.M., V.S. and A.F. did the NMR spectrometry analysis. J.X. and W.K.C. did the western blot assay and provided advice. M.S.P. designed and analyzed the PK studies. T.C. and J.M. helped with chemical synthesis. M.M.A., S.C.P. and M.R.M. wrote the manuscript, and all authors refined the manuscript.

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Correspondence to Mamoun M Alhamadsheh.

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Supplementary Results, Supplementary Table 1, Supplementary Figures 1–10 and Supplementary Notes 1 and 2 (PDF 3752 kb)

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Penchala, S., Miller, M., Pal, A. et al. A biomimetic approach for enhancing the in vivo half-life of peptides. Nat Chem Biol 11, 793–798 (2015). https://doi.org/10.1038/nchembio.1907

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