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The Use of Surfactants to Solubilise a Glucagon Analogue

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ABSTRACT

Purpose

The peptide hormone glucagon, used to treat hypoglycaemic incidents, is prone to aggregation. Generating alternatives with better stability is of pharmaceutical interest in the treatment of diabetes. Here we investigate the impact of six different surfactants on the solubility and stability of ZP-GA-1, a stable version of glucagon.

Methods

We use chemical surfactants (sodium dodecyl sulphate, dodecyl maltoside and polysorbate 20) and the biosurfactants rhamnolipid, sophorolipid and surfactin. We investigate their interaction with ZP-GA-1 by pyrene fluorescence, circular dichroism and isothermal titration calorimetry.

Results

All six surfactants induce α-helical structure in ZP-GA-1, SDS having the biggest impact and polysorbate 20 the smallest. SDS keeps ZP-GA-1 solubilised over >48 days as opposed to 29 days in DDM, 3 days in polysorbate 20 and 0 days in buffer. Similarly, much less SDS than DDM, polysorbate 20 or biosurfactant is needed to redissolve aggregated ZP-GA-1. ITC confirms this trend, with SDS exhibiting very strong, and polysorbate 20 very weak interactions.

Conclusion

Simple surfactant structures promote stronger peptide interactions. ITC shows promise as a general strategy to predict surfactants’ solubilising powers. Stronger enthalpic interactions improved the absolute solubility of ZP-GA-1 and their strength correlated to the absolute solubility of the peptides though not to the kinetics of precipitation.

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Abbreviations

DDM:

Dodecyl maltoside

Ps20:

Polysorbate 20

RL:

Rhamnolipid

SDS:

Sodium dodecyl sulphate

SF:

Surfactin

SL:

Sophorolipid

ZP-GA-1:

Glucagon analogue

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Acknowledgments and Disclosures

J.K.M. is funded by an Industrial PhD fellowship cofunded by Zealand Pharma A/S and the Danish Innovation Foundation (grant no. 1355–00136).

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Authors and Affiliations

  1. Interdisciplinary Nanoscience Centre (iNANO) and Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 14, DK 8000, Aarhus C, Denmark

    Jens Kvist Madsen & Daniel E. Otzen

  2. Department of Pharmaceutical Development, Zealand Pharma A/S, Smedeland 36, DK–Glostrup, Denmark

    Jens Kvist Madsen & Lise Giehm

Authors
  1. Jens Kvist Madsen
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  2. Lise Giehm
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  3. Daniel E. Otzen
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Correspondence to Lise Giehm or Daniel E. Otzen.

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Madsen, J.K., Giehm, L. & Otzen, D.E. The Use of Surfactants to Solubilise a Glucagon Analogue. Pharm Res 35, 235 (2018). https://doi.org/10.1007/s11095-018-2494-2

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