Abstract

One of the most significant challenges in the development of clinically viable delivery systems for RNA interference therapeutics is to understand how molecular structures influence delivery efficacy. Here, we have synthesized 1,400 degradable lipidoids and evaluate their transfection ability and structure–function activity. We show that lipidoid nanoparticles mediate potent gene knockdown in hepatocytes and immune cell populations on IV administration to mice (siRNA EC50 values as low as 0.01 mg kg−1). We identify four necessary and sufficient structural and pKa criteria that robustly predict the ability of nanoparticles to mediate greater than 95% protein silencing in vivo. Because these efficacy criteria can be dictated through chemical design, this discovery could eliminate our dependence on time-consuming and expensive cell culture assays and animal testing. Herein, we identify promising degradable lipidoids and describe new design criteria that reliably predict in vivo siRNA delivery efficacy without any prior biological testing.

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Acknowledgements

K.A.W. would like to thank D. Siegwart and C. Alabi for helpful discussions as well as F. Niroui and M. Ma for technical assistance. This work was funded by Alnylam Pharmaceuticals and NIH grants EB000244, R01CA115527 and R01CA132091. K.A.W. was supported by an NIH F32 fellowship (award number EB009623). O.V. acknowledges support from CDMRP, Department of Defense, postdoctoral fellowship award (#W81XWH-13-1-0215).

Author information

Author notes

    • Kathryn A. Whitehead

    Present address: Department of Chemical Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh, Pennsylvania 15217, USA

Affiliations

  1. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA

    • Kathryn A. Whitehead
    • , Arturo J. Vegas
    • , Philip H. Chang
    • , Omid Veiseh
    • , Jonathan Matthews
    • , Yunlong Zhang
    • , Karsten T. Olejnik
    • , Volkan Yesilyurt
    • , Delai Chen
    • , Robert Langer
    •  & Daniel G. Anderson
  2. Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA

    • J. Robert Dorkin
  3. Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA

    • Owen S. Fenton
  4. Alnylam Pharmaceuticals, 300 Third Street, Cambridge, Massachusetts 02142, USA

    • Scott Barros
    • , Boris Klebanov
    •  & Tatiana Novobrantseva
  5. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA

    • Robert Langer
    •  & Daniel G. Anderson
  6. The Institute for Medical Engineering and Science, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139 USA

    • Robert Langer
    •  & Daniel G. Anderson

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Contributions

K.A.W. designed and performed experiments, analysed data and wrote the paper. J.R.D., P.H.C., J.M., O.V., O.F., Y.Z., K.T.O., V.Y. and B.K. designed and performed experiments. D.C. designed and fabricated microfluidic devices. A.J.V., S.B. and T.N. provided conceptual advice and technical support. R.L. and D.G.A. supervised the study. All authors discussed the results and commented on the manuscript.

Competing interests

R.L. is a shareholder and member of the scientific advisory board of Alnylam. D.G.A. is a consultant with Alnylam Pharmaceuticals. R.L. and D.G.A. have sponsored research grants from Alnylam. Alnylam also has a license to certain intellectual property invented at Massachusetts Institute of Technology by K.A.W., J.R.D., A.J.V., R.L., and D.G.A. The remaining authors declare no competing financial interest.

Corresponding author

Correspondence to Daniel G. Anderson.

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DOI

https://doi.org/10.1038/ncomms5277

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