Сборник V международной

SYNTHESIS AND IN VITRO ANTITUMOR PROPERTY OF DOXORUBICIN 
CONJUGATED HYALURONAN NANOPARTICLES
Bekmirzayev J.N.1, Muhitdinov B.I.1, Turaev A.S.1, Huang Y.2, Sindarov B.A.1,
Wang H.2, Amonova D.M.1, Kirgizbayev H.H.1
1 Institute o f Bioorganic Chemistry, Uzbekistan Academy o f Sciences, 
2Shanghai Institute o f Materia Medica, Chinese Academy o f Sciences,
501 Hai Ke Road, 201203 Shanghai, China 
e-mail: [e-mail], [phone]
Recently studies have indicated that nano drug delivery technologies can be a promising approach 
for cancer treatment through targeted delivery o f therapeutic materials with reduced adverse effects. 
Biopolymers-based nanoplatforms have aroused great interest in cancer-specific drug delivery with respect 
to their biodegradable and biocompatible features. Natural polysaccharides such as chitosan, pectin, starch, 
cellulose, and hyaluronic acid abundantly occurring biopolymers have been intensively studied in cancer 
drug delivery. Hyaluronic acid (HA) is nonsulfated, linearly structured, and negatively charged glycosami- 
noglycan consisting of alternately repeated A-acetylglucosamine and glucuronic disaccharide. HA mac­
romolecule contains reactive functional groups for the synthesis o f modified derivatives. In addition, the 
biopolymer is a major component o f the extracellular matrix and plays a crucial role in the organism. Most 
importantly, along with biocompatible and biodegradable properties, the biopolymer specifically binds with 
HA-binding proteins frequently upregulated on malignant cells and the property has been used as the main 
tool for tumor drug delivery. To date, several approaches have been developed for preparing HA based 
nanoformulations for drug delivery.
In this study, we constructed a novel amphiphilic constitution on the HA chain by covalently attach­
ing hydrophobic palmitic acid (PA) and doxorubicin (DOX) as a model drug, using chemical conjugation 
methods. In the macromolecular system prepared, the HA-PA attachment was achieved using esterification 
reaction and the DOX conjugation was gained by peptide bond formation between the biopolymer and 
drug. The modified polysaccharide then nanoformulated into HA-PA-DOX nanoparticles by using self-as­
sembling methods. The nanoparticles prepared were purified by dialysis against distilled water to remove 
low molecular weight molecules. The drug loading capacity (12.48%, wt.) o f the nanoparticles was evaluat­
ed by UV spectroscopic method. In the IR spectroscopic studies, the nanoparticles were exhibited, the sig­
nals corresponding to HA glycoside bonds at 894-1060 cm-1. The presence of the DOX benzyl group in the 
samples was approved by a specific peak at 1454-1646 cm-1. The peaks appeared at 1600-1700 cm-1 showed 
HA-DOX peptide bond formation in the nanoparticles. The HA-PA ester bond formation was confirmed 
by the peak from ester C=O detected at 1643 cm-1. The dynamic light scattering measurements indicated 
that nanoparticles prepared were 201 nm by size and 0.16 by size distribution. The stable spherical-shaped 
nanoparticles were observed by the morphological studies by TEM, and the method confirmed nanoparti­
cle sizes similar to that detected by the dynamic light scattering method. Cytotoxicity o f the HA-PA-DOX 
nanoparticles against HeLa cells was evaluated by the MTT assay. The nanoparticles exhibited antitumor 
activity of 67.7%, while the free drug inhibited the cell growth by 64.2% at the same concentration (100 
hg/ml).
In summary, HA-PA-DOX nanoparticles with spherical-shaped morphological structure and an aver­
age size of 201 nm (size distribution 0.16) were prepared for tumor drug delivery. The prepared nanoparti­
cles demonstrated higher antitumor activity than that of the free drug at the same concentration against the 
HeLa cells. The nanoparticles are under study with respect to their detailed pharmacokinetic properties and 
in vivo antitumor activity.
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