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Selection of P-Glycoprotein Inhibitor and Formulation of Combinational Nanoformulation Containing Selected Agent Curcumin and DOX for Reversal of Resistance in K562 Cells

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Abstract

Purpose

To select P-glycoprotein (P-gp) inhibitor from natural source for reversal of DOX resistance in K562 cells and to develop selected one in to nanoformulation in combination with DOX.

Methods

DOX resistant K562 (K562R) cells were developed and reversal of resistance by P-gp inhibitor was validated by co-treatment with verapamil. The p-gp inhibitors were evaluated for their potential to inhibit P-gp (calcein assay) and to reverse drug resistance (XTT cell viability assay). The selected agent, curcumin was formulated in to liposome along with DOX and characterized for size, zeta potential, encapsulation efficiency and release rate. Uptake, P-gp inhibition and reversal of acquired drug resistance in K562R cells were performed.

Results

P-gp inhibitors such as biochanin-A and curcumin were marked suitable for combination with DOX. However, only curcumin could increase the sensitivity of DOX at all dosing levels, therefore used for further studies. Liposomes loaded with curcumin were formulated and characterized where a prolonged release was observed. The uptake of liposomal curcumin was comparable to nanodispersed curcumin but had lower cytotoxicity. DOX and curcumin coloaded liposomes successfully reversed DOX resistance in K562 cells. Conclusion: The coloaded liposomes increased the safety of curcumin with improved efficacy thus can be employed for reversal of acquired DOX resistance.

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Abbreviations

CAN:

Acetonitrile

BioA:

Biochanin A

Cur:

Curcumin

Daid:

Daidzein

DHF:

Dihydrofisetin

DMSO:

Dimethyl sulphoxide

DOX:

Doxorubicin

DSPE-PEG:

1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)]

EE:

Encapsulation efficiency

HSPC:

Hydrogenated soy phosphatidyl choline

MFI:

Mean fluorescent intensities

OPA:

Ortho-phosphoric acid

PFA:

Paraformal dehyde

P-gp:

P-glycoprotein

EPR:

Enhanced permeation and retention

Gen:

Genistein

mPEG:

Methoxy polyethyleneglycol

PCL:

Poly-ε- caprolactone

Resv:

Resveratrol

Sily:

Silymarin

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Acknowledgments and Disclosures

Authors acknowledge SERB, Govt. of India, for funding “Fast Track Scheme for Young Scientists” grant (No. SR/FT/LS-136/2011) and NISER, DAE, Govt. of India, for financial support as well as fellowship.

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Authors and Affiliations

  1. School of Biological Sciences, National Institute of Science Education and Research (NISER), Homi Bhabha National Institute (HBNI), PO- Bhimpur-Padanpur, Via- Jatni, Khurda, 752050, India

    Tapan K. Dash & V. Badireenath Konkimalla

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  1. Tapan K. Dash
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  2. V. Badireenath Konkimalla
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Correspondence to V. Badireenath Konkimalla.

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Dash, T.K., Konkimalla, V. Selection of P-Glycoprotein Inhibitor and Formulation of Combinational Nanoformulation Containing Selected Agent Curcumin and DOX for Reversal of Resistance in K562 Cells. Pharm Res 34, 1741–1750 (2017). https://doi.org/10.1007/s11095-017-2182-7

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