Journal of the American Chemical Society
ACKNOWLEDGMENTS
Page 4 of 5
nude mice bearing MDA-MB-231-Luc xenograft tumors in both
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flanks. After 2.5 hours, remarkably high tumor accumulation of
N1C4 (2:2) NPs was measured (Figure 5A). Fluorescence signals
from the liver and kidneys were also visualized. Ex vivo imaging of
harvested organs confirmed effective tumor uptake (Figure S42).
Moreover, luciferase activity in the tumors was greatly reduced
after intratumoral (IT) injection of 2.5 mg/kg siLuc. Luciferase was
quantified by bioluminescence (Figure 5B) and by tissue
homogenization on total protein and tissue levels (Figure 5C).
We gratefully acknowledge financial support from the Cancer
Prevention and Research Institute of Texas (CPRIT) Grant R1212
and the Welch Foundation Grant I-1855.
REFERENCES
(1) a) Tian, H. Y.; Tang, Z. H.; Zhuang, X. L.; Chen, X. S.; Jing, X. B.
Prog. Polym. Sci. 2012, 37, 237; b) Jerome, C.; Lecomte, P. Adv. Drug
Deliv. Rev. 2008, 60, 1056; c) Pounder, R.; Dove, A. Polym. Chem. 2010,
1, 260; d) Seyednejad, H.; Ghassemi, A. H.; van Nostrum, C. F.;
Vermonden, T.; Hennink, W. E. J. Controlled Release 2011, 152, 168; e)
Hao, J.; Rainbolt, E. A.; Washington, K.; Biewer, M. C.; Stefan, M. C.
Curr. Org. Chem. 2013, 17, 930.
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
(2) a) Pack, D. W.; Hoffman, A. S.; Pun, S.; Stayton, P. S. Nat. Rev. Drug
Discov. 2005, 4, 581; b) Whitehead, K.; Langer, R.; Anderson, D. Nat.
Rev. Drug Discov. 2009, 8, 129; c) Kanasty, R.; Dorkin, J. R.; Vegas, A.;
Anderson, D. Nat. Mater. 2013, 12, 967.
(3) Albertsson, A. C.; Varma, I. K. Adv. Polym. Sci. 2002, 157, 1.
(4) a) Akinc, A.; Zumbuehl, A.; Goldberg, M.; Leshchiner, E. S.; Busini,
V.; Hossain, N.; Bacallado, S. A.; Nguyen, D. N.; Fuller, J.; Alvarez, R.;
Borodovsky, A.; Borland, T.; Constien, R.; de Fougerolles, A.; Dorkin, J.
R.; Narayanannair Jayaprakash, K.; Jayaraman, M.; John, M.;
Koteliansky, V.; Manoharan, M.; Nechev, L.; Qin, J.; Racie, T.;
Raitcheva, D.; Rajeev, K. G.; Sah, D. W.; Soutschek, J.; Toudjarska, I.;
Vornlocher, H. P.; Zimmermann, T. S.; Langer, R.; Anderson, D. G. Nat.
Biotechnol. 2008, 26, 561; b) Siegwart, D. J.; Whitehead, K. A.; Nuhn, L.;
Sahay, G.; Cheng, H.; Jiang, S.; Ma, M. L.; Lytton-Jean, A.; Vegas, A.;
Fenton, P.; Levins, C. G.; Love, K. T.; Lee, H.; Cortez, C.; Collins, S. P.;
Li, Y. F.; Jang, J.; Querbes, W.; Zurenko, C.; Novobrantseva, T.; Langer,
R.; Anderson, D. G. Proc. Natl. Acad. Sci. U.S.A. 2011, 108, 12996; c)
Jayaraman, M.; Ansell, S. M.; Mui, B. L.; Tam, Y. K.; Chen, J. X.; Du, X.
Y.; Butler, D.; Eltepu, L.; Matsuda, S.; Narayanannair, J. K.; Rajeev, K.
G.; Hafez, I. M.; Akinc, A.; Maier, M. A.; Tracy, M. A.; Cullis, P. R.;
Madden, T. D.; Manoharan, M.; Hope, M. J. Angew. Chem. Int. Ed. 2012,
Figure 5. (A) N1C4 (2:2) NPs provided effective accumulation in
tumor xenografts after IV injection. A representative mouse is
shown from three angles. Luciferase silencing was measured in
tumors 24 hours after injection by (B) bioluminescence imaging or
(
C) in tissue lysates normalized against total protein level or total
tissue amount (n=4; *P < 0.05).
5
1, 8529; d) Scholz, C.; Wagner, E. J. Controlled Release 2012, 161, 554.
In the spectrum of delivery systems, polymers have many
advantages including tunable structural composition, degradability,
and biocompatibility. Yet, they are currently less effective than
lipid-based delivery vehicles. To overcome this challenge, we have
incorporated key ionizable amines and hydrophobic alkyl chains
into polyesters. We synthesized a library of lipocationic polyesters
directly from functional monomers in high yield, fast time (~2
minutes), and in gram scale. This was accomplished with precise
monomer incorporation ratios to enable tunable hydrophobicity and
pKa. Formulated NPs enabled siRNA mediated silencing in vitro
and in vivo at low does. Notably, NPs could localize to tumors in
vivo after IV delivery and were able to silence gene expression in
tumor-bearing mice. This new class of lipocationic polyesters is a
promising step towards closing the activity gap between lipids and
polymers. Finally, we envision that the versatility of the chemical
methods may allow preparation of functional polyesters for a
variety of applications (not only for gene delivery) because nearly
any thiol can be used to synthesize functional monomers.
(5) a) Lynn, D. M.; Langer, R. J. Am. Chem. Soc. 2000, 122, 10761; b)
Green, J.; Langer, R.; Anderson, D. Acc. Chem. Res. 2008, 41, 749; c)
Gokhale, S.; Xu, Y.; Joy, A. Biomacromolecules 2013, 14, 2489; d) Yan,
Y. F.; Siegwart, D. J. Polym. Chem. 2014, 5, 1362.
(6) a) van der Ende, A.; Kravitz, E.; Harth, E. J. Am. Chem. Soc. 2008,
130, 8706; b) Nederberg, F.; Zhang, Y.; Tan, J. P. K.; Xu, K. J.; Wang, H.
Y.; Yang, C.; Gao, S. J.; Guo, X. D.; Fukushima, K.; Li, L. J.; Hedrick, J.
L.; Yang, Y. Y. Nat. Chem. 2011, 3, 409; c) Zhang, X. J.; Cai, M. M.;
Zhong, Z. L.; Zhuo, R. X. Macromol. Rapid Comm. 2012, 33, 693; d)
Darcos, V.; Antoniacomi, S.; Paniagua, C.; Coudane, J. Polym. Chem.
2
012, 3, 362; e) Zhang, Z. H.; Yin, L. C.; Xu, Y. X.; Tong, R.; Lu, Y. B.;
Ren, J.; Cheng, J. J. Biomacromolecules 2012, 13, 3456; f) Kim, H.;
Olsson, J. V.; Hedrick, J. L.; Waymouth, R. M. ACS Macro Lett. 2012, 1,
8
Edward, J. A.; Hickerson, R. P.; Kaspar, R. L.; Hedrick, J. L.; Waymouth,
R. M.; Wender, P. A. Proc. Natl. Acad. Sci. U.S.A. 2012, 109, 13171; h)
Jones, C. H.; Chen, C. K.; Jiang, M.; Fang, L.; Cheng, C.; Pfeifer, B. A.
Mol. Pharmaceutics 2013, 10, 1138; i) Winkler, M.; Raupp, Y. S.; Kohl,
L. A. M.; Wagner, H. E.; Meier, M. A. R. Macromolecules 2014, 47,
45; g) Geihe, E. I.; Cooley, C. B.; Simon, J. R.; Kiesewetter, M. K.;
2
9
842; j) Blake, T. R.; Waymouth, R. M. J. Am. Chem. Soc. 2014, 136,
252.
ASSOCIATED CONTENT
Supporting Information
(7) a) Love, K.; Mahon, K.; Levins, C.; Whitehead, K.; Querbes, W.;
Dorkin, J.; Qin, J.; Cantley, W.; Qin, L.; Racie, T.; Frank-Kamenetsky,
M.; Yip, K.; Alvarez, R.; Sah, D.; de Fougerolles, A.; Fitzgerald, K.;
Koteliansky, V.; Akinc, A.; Langer, R.; Anderson, D. Proc. Natl. Acad.
Sci. U.S.A. 2010, 107, 1864; b) Nelson, C. E.; Kintzing, J. R.; Hanna, A.;
Shannon, J. M.; Gupta, M. K.; Duvall, C. L. ACS Nano 2013, 7, 8870.
(8) Sutar, A. K.; Maharana, T.; Dutta, S.; Chen, C. T.; Lin, C. C. Chem
Soc Rev 2010, 39, 1724.
Includes detailed Synthetic Procedures for all aminothiols,
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monomers, and polymers (including characterization by H and
C
NMR, LC-MS, and GPC) (Figures S1-S37), Materials,
Instrumentation, and Biological Studies (Figures S38-S43). This
information is available free of charge via the Internet at
(9) a) Tempelaar, S.; Mespouille, L.; Dubois, P.; Dove, A. P.
Macromolecules 2011, 44, 2084; b) Silvers, A. L.; Chang, C. C.; Emrick,
T. J. Polym. Sci., Part A: Polym. Chem. 2012, 50, 3517.
(10) Leung, A. K. K.; Hafez, I. M.; Baoukina, S.; Belliveau, N. M.;
AUTHOR INFORMATION
Corresponding Author
Zhigaltsev, I. V.; Afshinmanesh, E.; Tieleman, D. P.; Hansen, C. L.;
Hope, M. J.; Cullis, P. R. J. Phys. Chem. C 2012, 116, 18440.
*E-mail: daniel.siegwart@utsouthwestern.edu
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