Roman Akasov
1,2*, Maria Drozdova
1,3, Daria Zaytseva-Zotova
1, Maria Leko
4, Pavel Chelushkin
4, Annie Marc
5, Isabelle Chevalot
5, Sergey Burov
4, Natalia Klyachko
6, Thierry Vandamme
7, Elena Markvicheva
11 Polymers for Biology Laboratory, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 117997, Miklukho-Maklaya 16/10, Moscow, Russia.
2 Institute of Molecular Medicine, Sechenov First Moscow State Medical University, 119991, Trubetskaya str. 8-2, Moscow, Russia.
3 Institute for Regenerative Medicine, Sechenov First Moscow State Medical University, 119991, Trubetskaya str. 8-2, Moscow, Russia.
4 Synthesis of Peptides and Polymer Microspheres Laboratory, Institute of Macromolecular Compounds of the Russian Academy of Sciences, 199004, Bolshoi pr. 31, Saint-Petersburg, Russia.
5 UMR CNRS 7274, Laboratoire Réactions et Génie des Procédés, Université de Lorraine, 54518, 2 avenue de la Fort de Haye, Vandoeuvre lès Nancy, France.
6 Faculty of Chemistry, Lomonosov Moscow State University, 119991, Leninskiye Gory 1-3, Moscow, Russia.
7 CNRS UMR 7199, Laboratoire de Conception et Application de Molécules Bioactives, University of Strasbourg, 74 route du Rhin, 67401 Illkirch Cedex, France.
Abstract
Purpose: Multidrug resistance (MDR) of tumors to
chemotherapeutics often leads to failure of cancer treatment. The aim of the
study was to prepare novel MDR-overcoming chemotherapeutics based on
doxorubicin (DOX) derivatives and to evaluate their efficacy in 2D and 3D in vitro models.
Methods: To overcome MDR, we synthesized five DOX derivatives,
and then obtained non-covalent complexes with human serum albumin (HSA). Drug
efficacy was evaluated for two tumor cell lines, namely human breast
adenocarcinoma MCF-7 cells and DOX resistant MCF-7/ADR cells. Additionally,
MCF-7 cells were entrapped in alginate-oligochitosan microcapsules, and
generated tumor spheroids were used as a 3D in
vitro model to study cytotoxicity of the DOX derivatives.
Results: Due to 3D structure, the tumor spheroids were more
resistant to chemotherapy compared to monolayer culture. DOX covalently
attached to palmitic acid through hydrazone linkage (DOX-N2H-Palm
conjugate) was found to be the most promising derivative. Its accumulation
levels within MCF-7/ADR cells was 4- and 10-fold higher than those of native
DOX when the conjugate was added to cultivation medium without serum and to
medium supplemented with 10% fetal bovine serum, respectively. Non-covalent
complex of the conjugate with HSA was found to reduce the IC50 value
from 32.9 µM (for free DOX-N2H-Palm) to 16.8 µM (for HSA-DOX-N2H-Palm)
after 72 h incubation with MCF-7/ADR cells.
Conclusion: Palm-N2H-DOX conjugate was found to be
the most promising DOX derivative in this research. The formation of
non-covalent complex of Palm-N2H-DOX conjugate with HSA allowed
improving its anti-proliferative activity against both MCF-7 and MCF-7/ADR
cells.