Combinatorial synthesis of chemically diverse core-shell nanoparticles for intracellular delivery
Edited* by Alexander M. Klibanov, Massachusetts Institute of Technology, Cambridge, MA, and approved June 20, 2011 (received for review May 2, 2011)
Abstract
Analogous to an assembly line, we employed a modular design for the high-throughput study of 1,536 structurally distinct nanoparticles with cationic cores and variable shells. This enabled elucidation of complexation, internalization, and delivery trends that could only be learned through evaluation of a large library. Using robotic automation, epoxide-functionalized block polymers were combinatorially cross-linked with a diverse library of amines, followed by measurement of molecular weight, diameter, RNA complexation, cellular internalization, and in vitro siRNA and pDNA delivery. Analysis revealed structure-function relationships and beneficial design guidelines, including a higher reactive block weight fraction, stoichiometric equivalence between epoxides and amines, and thin hydrophilic shells. Cross-linkers optimally possessed tertiary dimethylamine or piperazine groups and potential buffering capacity. Covalent cholesterol attachment allowed for transfection in vivo to liver hepatocytes in mice. The ability to tune the chemical nature of the core and shell may afford utility of these materials in additional applications.
Acknowledgments.
The authors thank Boris Klebanov, Manos Karagiannis, Christian J. Kastrup, Avi Schroeder, Carmen Barnes, and Robert S. Siegwart for their help. This work was supported by Alnylam Pharmaceuticals and National Institutes of Health (NIH) Grants R01-EB000244-27 and 5-R01-CA132091-04. D.J.S. acknowledges postdoctoral support from NIH NRSA award F32-EB011867.
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References
1
A Balazs, T Emrick, T Russell, Nanoparticle polymer composites: Where two small worlds meet. Science 314, 1107–1110 (2006).
2
T Taton, C Mirkin, R Letsinger, Scanometric DNA array detection with nanoparticle probes. Science 289, 1757–1760 (2000).
3
A Shipway, E Katz, I Willner, Nanoparticle arrays on surfaces for electronic, optical, and sensor applications. ChemPhysChem 1, 18–52 (2000).
4
D LaVan, D Lynn, R Langer, Moving smaller in drug discovery and delivery. Nat Rev Drug Discov 1, 77–84 (2002).
5
ME Davis, Z Chen, DM Shin, Nanoparticle therapeutics: An emerging treatment modality for cancer. Nat Rev Drug Discov 7, 771–782 (2008).
6
H Geysen, F Schoenen, D Wagner, R Wagner, Combinatorial compound libraries for drug discovery: An ongoing challenge. Nat Rev Drug Discov 2, 222–230 (2003).
7
S Brocchini, K James, V Tangpasuthadol, J Kohn, A combinatorial approach for polymer design. J Am Chem Soc 119, 4553–4554 (1997).
8
T Boussie, et al., A fully integrated high-throughput screening methodology for the discovery of new polyolefin catalysts: Discovery of a new class of high temperature single-site group (IV) copolymerization catalysts. J Am Chem Soc 125, 4306–4317 (2003).
9
XD Xiang, et al., A combinatorial approach to materials discovery. Science 268, 1738–1740 (1995).
10
R Hoogenboom, M Meier, U Schubert, Combinatorial methods, automated synthesis and high-throughput screening in polymer research: Past and present. Macromol Rapid Commun 24, 16–32 (2003).
11
D Webster, Combinatorial and high-throughput methods in macromolecular materials research and development. Macromol Chem Phys 209, 237–246 (2008).
12
S Wong, N Sood, D Putnam, Combinatorial evaluation of cations, pH-sensitive and hydrophobic moieties for polymeric vector design. Mol Ther 17, 480–490 (2009).
13
AW Bosman, et al., High-throughput synthesis of nanoscale materials: Structural optimization of functionalized one-step star polymers. J Am Chem Soc 123, 6461–6462 (2001).
14
DG Anderson, DM Lynn, R Langer, Semi-automated synthesis and screening of a large library of degradable cationic polymers for gene delivery. Angew Chem Int Ed Engl 42, 3153–3158 (2003).
15
A Akinc, et al., A combinatorial library of lipid-like materials for delivery of RNAi therapeutics. Nat Biotechnol 26, 561–569 (2008).
16
A Hook, et al., High throughput methods applied in biomaterial development and discovery. Biomaterials 31, 187–198 (2010).
17
WA Braunecker, K Matyjaszewski, Controlled/living radical polymerization: Features, developments, and perspectives. Prog Polym Sci 32, 93–146 (2007).
18
G Moad, E Rizzardo, S Thang, Living radical polymerization by the RAFT process. Aust J Chem 58, 379–410 (2005).
19
H Zhang, V Marin, M Fijten, U Schubert, High-throughput experimentation in atom transfer radical polymerization: A general approach toward a directed design and understanding of optimal catalytic systems. J Polym Sci A Polym Chem 42, 1876–1885 (2004).
20
K Matyjaszewski, N Tsarevsky, Nanostructured functional materials prepared by atom transfer radical polymerization. Nat Chem 1, 276–288 (2009).
21
A Fire, et al., Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–811 (1998).
22
SM Elbashir, et al., Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411, 494–498 (2001).
23
K Whitehead, R Langer, D Anderson, Knocking down barriers: Advances in siRNA delivery. Nat Rev Drug Discov 8, 129–138 (2009).
24
K Love, et al., Lipid-like materials for low-dose, in vivo gene silencing. Proc Natl Acad Sci USA 107, 1864–1869 (2010).
25
DM Lynn, DG Anderson, D Putnam, R Langer, Accelerated discovery of synthetic transfection vectors: Parallel synthesis and screening of degradable polymer library. J Am Chem Soc 123, 8155–8156 (2001).
26
D Anderson, et al., A polymer library approach to suicide gene therapy for cancer. Proc Natl Acad Sci USA 101, 16028–16033 (2004).
27
J Green, R Langer, D Anderson, A combinatorial polymer library approach yields insight into nonviral gene delivery. Acc Chem Res 41, 749–759 (2008).
28
B Sun, X Liu, M Buck, D Lynn, Azlactone-functionalized polymers as reactive templates for parallel polymer synthesis: Synthesis and screening of a small library of cationic polymers in the context of DNA delivery. Chem Commun 46, 2016–2018 (2010).
29
A Nagayasu, K Uchiyama, H Kiwada, The size of liposomes: A factor which affects their targeting efficiency to tumors and therapeutic activity of liposomal antitumor drugs. Adv Drug Delivery Rev 40, 75–87 (1999).
30
G Zheng, C Pan, Reversible addition-fragmentation transfer polymerization in nanosized micelles formed in situ. Macromolecules 39, 95–102 (2006).
31
H Gao, K Matyjaszewski, Synthesis of functional polymers with controlled architecture by CRP of monomers in the presence of cross-linkers: From stars to gels. Prog Polym Sci 34, 317–350 (2009).
32
C Lee, J MacKay, J Frechet, F Szoka, Designing dendrimers for biological applications. Nat Biotechnol 23, 1517–1526 (2005).
33
M Davis, et al., Evidence of RNAi in humans from systemically administered siRNA via targeted nanoparticles. Nature 464, 1067–U1140 (2010).
34
D Chen, H Peng, M Jiang, A novel one-step approach to core-stabilized nanoparticles at high solid contents. Macromolecules 36, 2576–2578 (2003).
35
H Peng, D Chen, M Jiang, A one-pot approach to the preparation of organic core-shell nanoobjects with different morphologies. Macromolecules 38, 3550–3553 (2005).
36
HC Kolb, MG Finn, KB Sharpless, Click chemistry: Diverse chemical function from a few good reactions. Angew Chem Int Ed Engl 40, 2004–2021 (2001).
37
NV Tsarevsky, SA Bencherif, K Matyjaszewski, Graft copolymers by a combination of ATRP and two different consecutive click reactions. Macromolecules 40, 4439–4445 (2007).
38
A van der Ende, E Kravitz, E Harth, Approach to formation of multifunctional polyester particles in controlled nanoscopic dimensions. J Am Chem Soc 130, 8706–8713 (2008).
39
MD Kurkuri, et al., Multifunctional polymer coatings for cell microarray applications. Biomacromolecules 10, 1163–1172 (2009).
40
BS Kim, HF Gao, AA Argun, K Matyjaszewski, PT Hammond, All-star polymer multilayers as pH-responsive nanofilms. Macromolecules 42, 368–375 (2009).
41
H Gao, et al., Site isolation of emitters within cross-linked polymer nanoparticles for white electroluminescence. Nano Lett 10, 1440–1444 (2010).
42
M Stenzel, Hairy Core-Shell Nanoparticles via RAFT: Where are the opportunities and where are the problems and challenges? Macromol Rapid Commun 30, 1603–1624 (2009).
43
S Hecht, J Frechet, Dendritic encapsulation of function: Applying nature’s site isolation principle from biomimetics to materials science. Angew Chem Int Ed Engl 40, 74–91 (2001).
44
V Rodionov, et al., Easy access to a family of polymer catalysts from modular star polymers. J Am Chem Soc 132, 2570–2572 (2010).
45
J Oh, R Drumright, D Siegwart, K Matyjaszewski, The development of microgels/nanogels for drug delivery applications. Prog Polym Sci 33, 448–477 (2008).
46
R O’Reilly, C Hawker, K Wooley, Cross-linked block copolymer micelles: Functional nanostructures of great potential and versatility. Chem Soc Rev 35, 1068–1083 (2006).
47
K Zhang, H Fang, Z Wang, J Taylor, K Wooley, Cationic shell-crosslinked knedel-like nanoparticles for highly efficient gene and oligonucleotide transfection of mammalian cells. Biomaterials 30, 968–977 (2009).
48
S Matsumoto, et al., Environment-responsive block copolymer micelles with a disulfide cross-linked core for enhanced siRNA delivery. Biomacromolecules 10, 119–127 (2009).
49
X Yin, H Stover, Hydrogel microspheres by thermally induced coacervation of poly(N,N-dimethylacrylamide-co-glycidyl methacrylate) aqueous solutions. Macromolecules 36, 9817–9822 (2003).
50
D Suzuki, H Kawaguchi, Modification of gold nanoparticle composite nanostructures using thermosensitive core-shell particles as a template. Langmuir 21, 8175–8179 (2005).
51
N Hantzschel, F Zhang, F Eckert, A Pich, M Winnik, Poly(N-vinylcaprolactam-co-glycidyI methacrylate) aqueous microgels labeled with fluorescent LaF3 : Eu nanoparticles. Langmuir 23, 10793–10800 (2007).
52
C Lorenz, P Hadwiger, M John, H Vornlocher, C Unverzagt, Steroid and lipid conjugates of siRNAs to enhance cellular uptake and gene silencing in liver cells. Bioorg Med Chem Lett 14, 4975–4977 (2004).
53
C Wolfrum, et al., Mechanisms and optimization of in vivo delivery of lipophilic siRNAs. Nat Biotechnol 25, 1149–1157 (2007).
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Published online: July 22, 2011
Published in issue: August 9, 2011
Acknowledgments
The authors thank Boris Klebanov, Manos Karagiannis, Christian J. Kastrup, Avi Schroeder, Carmen Barnes, and Robert S. Siegwart for their help. This work was supported by Alnylam Pharmaceuticals and National Institutes of Health (NIH) Grants R01-EB000244-27 and 5-R01-CA132091-04. D.J.S. acknowledges postdoctoral support from NIH NRSA award F32-EB011867.
Notes
*This Direct Submission article had a prearranged editor.
Authors
Competing Interests
Conflict of interest statement: R.L. is a shareholder and member of the Scientific Advisory Board of Alnylam. D.G.A. is a consultant with Alnylam. R.L and D.G.A have sponsored research grants from Alnylam. Alnylam also has a license to certain intellectual property invented at MIT. W.Q., C.Z., and T.N. are employed by Alnylam.
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Combinatorial synthesis of chemically diverse core-shell nanoparticles for intracellular delivery, Proc. Natl. Acad. Sci. U.S.A.
108 (32) 12996-13001,
https://doi.org/10.1073/pnas.1106379108
(2011).
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