Synthesis of L-DOPA conjugated doxorubicin-polyethylenimine nanocarrier and evaluation of its cytotoxicity on A375 and HepG2 cell lines

Document Type : Research Paper

Authors

1 Pharmaceutical Sciences Research center, Shiraz University of Medical Sciences, Shiraz, Iran

2 Department of Pharmaceutics, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran

3 Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran

Abstract

Objective(s): Polyethylenimine (PEI) is one of the most-extensively investigated cationic polymers for gene and drug delivery. Recently, great attention has been directed to design of carriers for co-delivery of nucleic acids and small molecules. These delivery systems are able to overcome the limitations of gene or drug delivery alone. The aim of this study is to prepare a targeted nano-carrier for co-delivery of doxorubicin (Dox) and gene using polyethylenimine.
Materials and Methods: In order to prepare the ligand-containing polymer conjugates, succinic anhydride was conjugated onto the hydroxyl group of Dox through an ester bond following the protection of Dox amines by Fmoc. Drug-polymer conjugates were then coupled with L-DOPA in order to prepare the targeted nanocarriers to the cells through Large Amino Acid Transporter-1 (LAT-1). The PEI derivatives were characterized using 1H-NMR. The toxicity of conjugated polymer, Dox and PEI was assessed on HepG2 and A375 cell lines with different expression level of LAT-1 transporters using MTT assay.
Results: The chemical structure of PEI conjugate was confirmed by 1H-NMR. The cytotoxicity measurement demonstrated a cell line-dependent toxicity profile at the concentrations tested in this study. It was shown that there was no significant difference in cell-induced toxicity between conjugated polymer and its parent form in A375 cell line while the cytotoxicity of conjugated polymer was significantly lower than the parent PEI in HepG2 cells.
Conclusion: These results provide promising evidence for further evaluation of PEI conjugate for co-delivery of drug and gene via LAT-1 transporters.

Keywords

References

1.    Arcamone F. Doxorubicin: anticancer antibiotics. Elsevier; 2012.
2.    Carvalho C, Santos RX, Cardoso S, Correia S, Oliveira PJ, Santos MS, et al. Doxorubicin: the good, the bad and the ugly effect. Curr Med Chem. 2009;16(25):3267-3285.
3.    Yang Z, Gao D, Cao Z, Zhang C, Cheng D, Liu J, Shuai X. Drug and gene co-delivery systems for cancer treatment. Biomater Sci. 2015;3(7):1035-1049.
4.    Zakeri A, Kouhbanani MAJ, Beheshtkhoo N, Beigi V, Mousavi SM, Hashemi SA, Jahandideh S. Polyethylenimine-based nanocarriers in co-delivery of drug and gene: a developing horizon. Nano Rev Exp. 2018;9(1):1488497.
5.    Yin H, Kanasty RL, Eltoukhy AA, Vegas AJ, Dorkin JR, Anderson DG. Non-viral vectors for gene-based therapy. Nat Rev Genet. 2014; 15(8):541-555.
6.    Pollard H, Remy J-S, Loussouarn G, Demolombe S, Behr J-P, Escande D. Polyethylenimine but not cationic lipids promotes transgene delivery to the nucleus in mammalian cells. J Biol Chem. 1998;273(13):7507-7511.
7.    Oskuee RK, Dehshahri A, Shier WT, Ramezani M. Alkylcarboxylate grafting to polyethylenimine: a simple approach to producing a DNA nanocarrier with low toxicity. J Gene Med. 2009; 11(10):921-932.
8.    Dehshahri A, Sadeghpour H, Mohazzabieh E, Saatchi Avval S, Mohammadinejad R. Targeted double domain nanoplex based on galactosylated polyethylenimine enhanced the delivery of IL‐12 plasmid. Biotechnol prog. 2020;36(5):e3002.
9.    Häfliger P, Charles R-P. The L-type amino acid transporter LAT1—An emerging target in cancer. Int J Mol Sci. 2019;20(10):2428.
10.    Ong ZY, Chen S, Nabavi E, Regoutz A, Payne DJ, Elson DS, et al. Multibranched gold nanoparticles with intrinsic LAT-1 targeting capabilities for selective photothermal therapy of breast cancer. ACS Appl Mater  interfaces. 2017;9(45):39259-39270.
11.    Nagy A, Schally AV, Armatis P, Szepeshazi K, Halmos G, Kovacs M, Zarandi M, Groot K, Miazaki M, Jungwirth A, Horvath J. Cytotoxic analogs of luteinizing hormone-releasing hormone containing doxorubicin or 2-pyrrolinodoxorubicin, a derivative 500-1000 times more potent. Proc Natl Acad Sci. 1996 9;93(14):7269-7273.
12.    Dong DW, Tong SW, Qi XR. Comparative studies of polyethylenimine–doxorubicin conjugates with pH‐sensitive and pH‐insensitive linkers. J Biomed Mater Res A. 2013;101(5):1336-1344.
13.    Talebpour Z, Haghgoo S, Shamsipur M. 1H nuclear magnetic resonance spectroscopy analysis for simultaneous determination of levodopa, carbidopa and methyldopa in human serum and pharmaceutical formulations. Anal Chim Acta. 2004;506(1):97-104.
14.    Arcamone F. Structure-activity relationships in doxorubicin related compounds.  InStructure-Activity Relationships of Anti-Tumour Agents 1983 (pp.111-133). Springer,  Dordrecht.
15.    Zintchenko A, Philipp A, Dehshahri A, Wagner E. Simple modifications of branched PEI lead to highly efficient siRNA carriers with low toxicity. Bioconjug Chem. 2008;19(7):1448-1455.
16.    Dehshahri A, Sadeghpour H, Oskuee RK, Fadaei M, Sabahi Z, Alhashemi SH, Mohazabie E. Interleukin-12 plasmid DNA delivery using l-thyroxine-conjugated polyethylenimine nanocarriers. J Nanoparticle Res. 2014;16(5):1-14.
17.    Khalvati B, Sheikhsaran F, Sharifzadeh S, Kalantari T, Behzad Behbahani A, Jamshidzadeh A, Dehshahri A. Delivery of plasmid encoding interleukin-12 gene into hepatocytes by conjugated polyethylenimine-based nanoparticles. Artif Cells nanomed Biotechnol. 2017;45(5):1036-1044.
18.    Sadeghpour H, Khalvati B, Entezar-Almahdi E, Savadi N, Alhashemi SH, Raoufi M, Dehshahri A. Double domain polyethylenimine-based nanoparticles for integrin receptor mediated delivery of plasmid DNA. Sci Rep. 2018;8(1):1-12.
19.    Sheikhsaran F, Sadeghpour H, Khalvati B, Entezar-Almahdi E, Dehshahri A. Tetraiodothyroacetic acid-conjugated polyethylenimine for integrin receptor mediated delivery of the plasmid encoding IL-12 gene. Colloids Surf B. 2017;150:426-436.
Nanomedicine Journal
Volume 8, Issue 4
October 2021
Pages 264-269

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