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Folate-Mediated Targeted Delivery of Combination Chemotherapeutics Loaded Reduced Graphene Oxide for Synergistic Chemo-Photothermal Therapy of Cancers

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Abstract

Purpose

Larger surface area for drug incorporation and superior optical activity makes reduced graphene oxide (rGO) a suitable drug carrier for combination chemotherapeutics delivery. And folate receptors are potential mediators for cancer targeted delivery. This study mainly aimed to prepare irinotecan (IRI)- and docetaxel (DOC)-loaded, folate (FA)-conjugated rGO (FA-P407-rGO/ID) for synergistic cancer therapy.

Methods

FA-P407-rGO/ID was prepared as aqueous dispersion. Characterization was performed using high performance liquid chromatography (HPLC), transmission electron microscopy (TEM), atomic force microscopy (AFM), ultraviolet/visible spectroscopy, fourier transform infrared spectroscopy (FTIR) and drug release. In vitro cellular studies were performed using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS), fluorescence-activated cell sorting (FACS) and western blot analyses.

Results

Our results revealed successful preparation of stable FA-P407-rGO/ID formulation with enhanced drug release profiles in acidic microenvironment. In vitro cytotoxicity of the formulation on folate receptor-expressing human mammary carcinoma (MCF-7) cells was higher than that when free IRI/DOC combination (ID) was used; such increased cytotoxicity was not observed in folate receptor-negative hepatocellular carcinoma (HepG2) cells. Cellular uptake of FA-P407-rGO/ID in MCF-7 cells was higher than in HepG2 cells. Further, FACS and western blot analysis revealed better apoptotic effects of the formulation in MCF-7 cells than in HepG2 cells, suggesting the important role of folate receptors for targeted chemotherapy delivery to cancer cells. Near infrared irradiation further enhanced the apoptotic effect in cancer cells, resulting from the photothermal effects of rGO.

Conclusions

Hence, FA-P407-rGO/ID can be considered as a potential formulation for folate-targeted chemo-photothermal therapy in cancer cells.

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Abbreviations

1H-NMR:

Proton nuclear magnetic resonance

AFM:

Atomic force microscopy

CDI:

1,1ʹ-carbonyldiimidazole

DOC:

Docetaxel

FA:

Folic acid

FACS:

Fluorescence-activated cell sorting

FITC:

Fluorescein-5(6)-isothiocyanate

FTIR:

Fourier transform infrared spectroscopy

HPLC:

High performance liquid chromatography

ID:

Irinotecan and docetaxel combination

IRI:

Irinotecan

MTS:

3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium

NIR:

Near infrared

P407:

Poloxamer 407

rGO:

Reduced graphene oxide

TEM:

Transmission electron microscopy

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ACKNOWLEDGMENTS AND DISCLOSURES

This research was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (No. 2015R1A2A2A01004118, 2015R1A2A2A04004806). This work was also supported by the Medical Research Center Program (2015R1A5A2009124) through the NRF funded by MSIP.

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Authors and Affiliations

  1. College of Pharmacy, Yeungnam University, 280 Daehak-Ro, Gyeongsan, 712-749, South Korea

    Raj Kumar Thapa, Yongjoo Choi, Jee-Heon Jeong, Chul Soon Yong & Jong Oh Kim

  2. School of Pharmacy, SungKyunKwan University, 300 Cheoncheon-dong, Jangan-gu, Suwon, 440-746, South Korea

    Yu Seok Youn

  3. College of Pharmacy, Hanyang University, 55, Hanyangdaehak-ro, Sangnok-gu, Ansan, 426-791, South Korea

    Han-Gon Choi

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  1. Raj Kumar Thapa
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Correspondence to Chul Soon Yong or Jong Oh Kim.

Electronic supplementary material

Characterizations of FA-P407 conjugate and FA-P407-rGO/ID; cytotoxicity and cellular uptake analysis.

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Thapa, R.K., Choi, Y., Jeong, JH. et al. Folate-Mediated Targeted Delivery of Combination Chemotherapeutics Loaded Reduced Graphene Oxide for Synergistic Chemo-Photothermal Therapy of Cancers. Pharm Res 33, 2815–2827 (2016). https://doi.org/10.1007/s11095-016-2007-0

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