10.1002/anie.202107221
Angewandte Chemie International Edition
COMMUNICATION
2016, 57, 943–949.
(Figure 4b), and that both targeting and high drug loading is
necessary in the tested setup to see lowered cell viability.
[6]
[7]
A. Younes, N. L. Bartlett, J. P. Leonard, D. A. Kennedy, C. M.
Lynch, E. L. Sievers, A. Forero-Torres, N. Engl. J. Med. 2010, 363,
1812–1821.
S. Yaghoubi, M. H. Karimi, M. Lotfinia, T. Gharibi, M. Mahi-
Birjand, E. Kavi, F. Hosseini, K. Sineh Sepehr, M. Khatami, N.
Bagheri, M. Abdollahpour-Alitappeh, J. Cell. Physiol. 2020, 235,
31–64.
R. P. Lyon, J. R. Setter, T. D. Bovee, S. O. Doronina, J. H. Hunter,
M. E. Anderson, C. L. Balasubramanian, S. M. Duniho, C. I. Leiske,
F. Li, P. D. Senter, Nat. Biotechnol. 2014, 32, 1059–1062.
L. Wang, G. Amphlett, W. A. Blättler, J. M. Lambert, W. Zhang,
Protein Sci. 2005, 14, 2436–2446.
R. Y. Zhao, S. D. Wilhelm, C. Audette, G. Jones, B. A. Leece, A. C.
Lazar, V. S. Goldmacher, R. Singh, Y. Kovtun, W. C. Widdison, J.
M. Lambert, R. V. J. Chari, J. Med. Chem. 2011, 54, 3606–3623.
P. M. LoRusso, D. Weiss, E. Guardino, S. Girish, M. X. Sliwkowski,
Clin. Cancer Res. 2011, 17, 6437–6447.
J. R. Junutula, H. Raab, S. Clark, S. Bhakta, D. D. Leipold, S. Weir,
Y. Chen, M. Simpson, S. P. Tsai, M. S. Dennis, Y. Lu, Y. G. Meng, C.
Ng, J. Yang, C. C. Lee, E. Duenas, J. Gorrell, V. Katta, A. Kim, K.
McDorman, K. Flagella, R. Venook, S. Ross, S. D. Spencer, W. Lee
Wong, H. B. Lowman, R. Vandlen, M. X. Sliwkowski, R. H.
Scheller, P. Polakis, W. Mallet, Nat. Biotechnol. 2008, 26, 925–
932.
The construct developed herein would likely have a low in
vivo half-life as a result of degradation of the DNA nanostructure
used. To investigate this the serum stability of the final construct
was investigated in vitro, and it was found that the structure is
completely degraded over a 24 h period in 10% FBS, Fig. S11 in
the supplementary information. This would likely mean that if the
structure developed here was used in vivo untargeted release of
MMAE would occur. Another issue for the construct used here is
that its large scale production would likely be difficult. With this
study we hope to inspire the development and use of simpler
oligonucleotide nanostructures combined with antibodies, with
life-time prolonging modifications such as PEG chains or non-
natural oligonucleotides implemented.
[8]
[9]
[10]
[11]
[12]
To conclude, we have demonstrated a new approach to
generate an ADC with an exact number of drug molecules, in this
case seven copies of MMAE. Each of the drugs are covalently
linked to ssDNA strands via a cleavable linker. Seven of the
MMAE modified strands are assembled in a small DNA cube,
which was used as the drug carrier and easily attached to a
Trastuzumab-DNA
conjugate
by
hybridization.
DNA
[13]
[14]
Y. T. Adem, K. A. Schwarz, E. Duenas, T. W. Patapoff, W. J.
Galush, O. Esue, Bioconjug. Chem. 2014, 25, 656–664.
C. A. Boswell, E. E. Mundo, C. Zhang, D. Bumbaca, N. R. Valle, K.
R. Kozak, A. Fourie, J. Chuh, N. Koppada, O. Saad, H. Gill, B. Q.
Shen, B. Rubinfeld, J. Tibbitts, S. Kaur, F. P. Theil, P. J. Fielder, L.
A. Khawli, K. Lin, Bioconjug. Chem. 2011, 22, 1994–2004.
G. Falck, K. M. Müller, Antibodies 2018, 7, 4.
M. Madsen, K. V Gothelf, Chem. Rev. 2019, 6384–6458.
V. L. Andersen, M. Vinther, R. Kumar, A. Ries, J. Wengel, J. S.
Nielsen, J. Kjems, Theranostics 2019, 9, 2662–2677.
R. Hu, X. Zhang, Z. Zhao, G. Zhu, T. Chen, T. Fu, W. Tan, Angew.
Chemie - Int. Ed. 2014, 53, 5821–5826.
Y. X. Zhao, A. Shaw, X. Zeng, E. Benson, A. M. Nyström, B.
Högberg, ACS Nano 2012, 6, 8684–8691.
Q. Jiang, C. Song, J. Nangreave, X. Liu, L. Lin, D. Qiu, Z. G. Wang,
G. Zou, X. Liang, H. Yan, B. Ding, J. Am. Chem. Soc. 2012, 134,
13396–13403.
nanostructures are available in a plethora of different sizes and
shapes and offers great control and opportunities to vary the
number of drug molecules associated with the complex. Beside
the high control of drug loading, the structures also offer
advantages such as the charge of the oligonucleotides, which
serves to keep multiple copies of a hydrophobic drug such as
MMAE in solution. A dramatic decline in viability to approximately
40% is observed only for cells that express the HER2 receptor
and are exposed to the full conjugate prepared here. While this
approach provides proof-of-concept, some challenges of the
system must be addressed before this system can be applied in
vivo. Most importantly the DNA sequences must be modified or
replaced with nucleotide analogues to resist enzymatic
degradation and we will address this in future studies.
[15]
[16]
[17]
[18]
[19]
[20]
[21]
[22]
[23]
[24]
[25]
[26]
[27]
[28]
S. Bi, B. Xiu, J. Ye, Y. Dong, ACS Appl. Mater. Interfaces 2015, 7,
23310–23319.
T. Liu, P. Song, A. Märcher, J. Kjems, C. Yang, K. V. Gothelf,
ChemBioChem 2019, 20, 1014–1018.
N. S. Hsu, C. C. Lee, W. C. Kuo, Y. W. Chang, S. Y. Lo, A. H. J.
Wang, Bioconjug. Chem. 2020, 31, 1804–1811.
J. Fu, G. Stankeviciute, S. W. Oh, J. Collins, Y. Zhong, T. Zhang,
Curr. Top. Med. Chem. 2017, 17, 1815–1828.
J. W. Conway, C. K. Mc Laughlin, K. J. Castor, H. Sleiman, Chem.
Commun. 2013, 49, 1172–1174.
A. Alagia, M. Terrazas, R. Eritja, Molecules 2014, 19, 17872–
17896.
C. J. Serpell, T. G. W. Edwardson, P. Chidchob, K. M. M. Carneiro,
H. F. Sleiman, J. Am. Chem. Soc. 2014, 136, 15767–15774.
K. E. Bujold, J. Fakhoury, T. G. W. Edwardson, K. M. M. Carneiro,
J. N. Briard, A. G. Godin, L. Amrein, G. D. Hamblin, L. C. Panasci,
P. W. Wiseman, H. F. Sleiman, Chem. Sci. 2014, 5, 2449–2455.
T. Trinh, C. Liao, V. Toader, M. Barlóg, H. S. Bazzi, J. Li, H. F.
Sleiman, Nat. Chem. 2018, 10, 184–192.
A. Lacroix, E. Vengut-Climent, D. De Rochambeau, H. F. Sleiman,
ACS Cent. Sci. 2019, 5, 882–891.
Y. H. Tan, M. Liu, B. Nolting, J. G. Go, J. Gervay-hague, G. Liu, ACS
Acknowledgements
We would like to thank Professor Hanadi Sleiman for advice to
the design and assembly of the cube structure. We are grateful to
the Novo Nordic Foundation for support to Center for
Multifunctional Biomolecular Drug Design (Grant No.
NNF17OC0028070).
Conflict of interest
The authors declare no conflict of interest.
[29]
[30]
[31]
[32]
Keywords: antibody drug conjugate, controlled DAR, protein
conjugation, DNA nanotechnology, atomic force microscopy
[1]
[2]
[3]
D. M. Ecker, S. D. Jones, H. L. Levine, MAbs 2015, 7, 9–14.
B. Kelley, MAbs 2009, 1, 443–452.
A. A. Shukla, L. S. Wolfe, S. S. Mostafa, C. Norman, Bioeng.
Transl. Med. 2017, 2, 58–69.
Nano 2008, 2, 2374–2384.
P. J. Burke, J. Z. Hamilton, S. C. Jeffrey, J. H. Hunter, S. O.
Doronina, N. M. Okeley, J. B. Miyamoto, M. E. Anderson, I. J.
Stone, M. L. Ulrich, J. K. Simmons, E. E. McKinney, P. D. Senter, R.
P. Lyon, Mol. Cancer Ther. 2017, 16, 116–123.
F. S. Ekholm, S. K. Ruokonen, M. Redón, V. Pitkänen, A. Vilkman,
J. Saarinen, J. Helin, T. Satomaa, S. K. Wiedmer, Separations
[4]
[5]
M. X. Sliwkowski, I. Mellman, Science (80-. ). 2013, 341, 1192–
1198.
N. Krall, F. Pretto, M. Mattarella, C. Muller, D. Neri, J. Nucl. Med.
[33]
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