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Di-Peptide-Modified Gemini Surfactants as Gene Delivery Vectors: Exploring the Role of the Alkyl Tail in Their Physicochemical Behavior and Biological Activity

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Abstract

The aim of this work was to elucidate the structure-activity relationship of new peptide-modified gemini surfactant-based carriers. Glycyl-lysine modified gemini surfactants that differ in the length and degree of unsaturation of their alkyl tail were used to engineer DNA nano-assemblies. To probe the optimal nitrogen to phosphate (N/P) ratio in the presence of helper lipid, in vitro gene expression and cell toxicity measurements were carried out. Characterization of the nano-assemblies was accomplished by measuring the particle size and surface charge. Morphological characteristics and lipid organization were studied by small angle X-ray scattering technique. Lipid monolayers were studied using a Langmuir-Blodgett trough. The highest activity of glycyl-lysine modified gemini surfactants was observed with the 16-carbon tail compound at 2.5 N/P ratio, showing a 5- to 10-fold increase in the level of reporter protein compared to the 12 and 18:1 carbon tail compounds. This ratio is significantly lower compared to the previously studied gemini surfactants with alkyl or amino- spacers. In addition, the 16-carbon tail compound exhibited the highest cell viability (85%). This high efficiency is attributed to the lowest critical micelle concentration of the 16-tail gemini surfactant and a balanced packing of the nanoparticles by mixing a saturated and unsaturated lipid together. At the optimal N/P ratio, all nanoparticles exhibited an inverted hexagonal lipid assembly. The results show that the length and nature of the tail of the gemini surfactants play an important role in determining the transgene efficiency of the delivery system. We demonstrated here that the interplay between the headgroup and the nature of tail is specific to each series, thus in the process of rational design, the contribution of the latter should be assessed in the appropriate context.

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Acknowledgments

Mays Al-Dulaymi is a fellow of the Canadian Institutes of Health Research Training grant in Health Research Using Synchrotron Techniques (CIHR-THRUST) and thanks the program for financial support. The authors acknowledge Mr. McDonald Donkuru, Dr. M. Jake Pushie, Dr. Masoomeh Poorghorban, and Mr. Osama Alaidi for the assistance on software training and experimental runs. The SAXS work was carried out at the Stanford Synchrotron Radiation Lightsource (SSRL), a Directorate of SLAC National Accelerator Laboratory, and an Office of Science User Facility operated for the US Department of Energy (DOE) Office of Science by Stanford University. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research, the National Institutes of Health (NIH), and the National Institute of General Medical Sciences (NIGMS; including P41GM103393). The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH. We acknowledge the assistance of Dr. Thomas Weiss, SSRL, with instrument setting and data collection. We thank Dr. Matthew F. Paige for facilitating the access to the Langmuir minitrough system.

This work was funded by the Saskatchewan Health Research Foundation, Natural Sciences and Engineering Research Council and The College of Pharmacy and Nutrition, University of Saskatchewan.

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Authors and Affiliations

  1. College of Pharmacy and Nutrition, University of Saskatchewan, Saskatoon, Saskatchewan, Canada

    Mays A. Al-Dulaymi, Waleed Mohammed-Saeid, Pawel Grochulski & Ildiko Badea

  2. Department of Chemical and Biological Engineering, University of Saskatchewan, Saskatoon, Saskatchewan, Canada

    Jackson M. Chitanda

  3. Department of Chemistry, University of Saskatchewan, Saskatoon, Saskatchewan, Canada

    Hessamaddin Younesi Araghi & Ronald E. Verrall

  4. Canadian Light Source, Saskatoon, Saskatchewan, Canada

    Pawel Grochulski

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  1. Mays A. Al-Dulaymi
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Correspondence to Ildiko Badea.

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Al-Dulaymi, M.A., Chitanda, J.M., Mohammed-Saeid, W. et al. Di-Peptide-Modified Gemini Surfactants as Gene Delivery Vectors: Exploring the Role of the Alkyl Tail in Their Physicochemical Behavior and Biological Activity. AAPS J 18, 1168–1181 (2016). https://doi.org/10.1208/s12248-016-9906-1

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