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Molecular Targets of the Hydrophobic Block of Pluronics in Cells: a Photo Affinity Labelling Approach

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Abstract

Purpose

Pluronics are known as inhibitors of multidrug resistance thus making tumor cells sensitive to therapeutic doses of drugs. The purpose of our study consists in revealing molecular targets of the hydrophobic poly(propylene oxide) block of pluronics in living cells and the dependence of the polymers chemosensitizing efficiency upon targeting.

Methods

A photo sensitive tracer was attached to the hydrophobic poly(propylene oxide) block of 3H-labeled tert-Bu-EO-PO copolymer. The conjugate was used for treatment cells in culture. We searched for its complexes with cellular lipids or proteins using RP TLC and SDS-electrophoresis, respectively. The chemosensitizing efficiency of pluronics was evaluated by their least concentrations sufficient for MDR reversion (CMDR).

Results

The poly(propylene oxide) block inserts in the lipid core of plasma membrane. No preferential binding of the conjugate with any cellular protein, particularly P-gp, has been detected. FITC-labeled pluronic L61 bound to alcohol insoluble cellular targets did not participate in MDR reversion. CMDR values of 13 block copolymers have been determined. These values inversely correlated with the polymers affinity toward lipids and the ability to accelerate flip-flop.

Conclusion

Insertion of the hydrophobic poly(propylene oxide) block of amphiphiles in the lipid core of plasma membrane and acceleration of flip-flop of lipids underlie the mechanism of MDR reversion.

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Abbreviations

BSA:

Bovine serum albumin

CMDR :

The least polymer concentration sufficient for the maximum possible MDR inhibition

DMEM:

Dulbecco’s modified Eagle’s medium

DOX:

Doxorubicin

GPC:

Gel permeation chromatography

EO:

Ethylene oxide

FITC:

Fluorescein isothiocyanate

PAL:

Photo affinity labeling

PBS:

Dulbecco’s phosphate-buffered saline

PO:

Propylene oxide

PPO:

Poly(propylene oxide)

R18:

Octadecyl Rhodamine B chloride

REP:

Tert-butoxy-poly(ethylene oxide)-b-poly(propylene oxide) block copolymer

RP TLC:

Reverse phase TLC

TFMD:

p-1-trifluoromethyldiazirinyl benzoic acid

TLC:

Thin layer chromatography

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ACKNOWLEDGMENTS AND DISCLOSURES

The authors express their gratitude to Prof. G. A. Korshunova for her generous gift of p-1-trifluorometyldiazirinyl benzoic acid (TFMD) and to the Shared Research Center of MSU supported by the Russian Ministry of Education and Science for the equipment provided.

Funding

This work was partially supported by grant RFBR #18-03-01234.

Author information

Authors and Affiliations

  1. Department of Chemistry, Lomonosov Moscow State University, GSP-1, Leninskie gory 1, build. 3, Moscow, 119991, Russia

    A. Zhirnov, G. Badun, N. Melik-Nubarov & I. Grozdova

  2. Australian Institute for Bioengineering and Nanotechnology (AIBN), University of Queensland, Corner College and Cooper Rds. (Bldg. 75), Brisbane, QLD, 4072, Australia

    E. Nam

  3. Department of Chemistry, Aarhus University, 8000, Langelandsgade, 140, Aarhus, Denmark

    A. Romanyuk

  4. Faculty of Physics, Lomonosov Moscow State University, GSP-1, Leninskie gory 1, build. 2, Moscow, 119991, Russia

    A. Ezhov

Authors
  1. A. Zhirnov
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  2. E. Nam
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  3. G. Badun
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  6. N. Melik-Nubarov
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  7. I. Grozdova
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Corresponding author

Correspondence to I. Grozdova.

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Zhirnov, A., Nam, E., Badun, G. et al. Molecular Targets of the Hydrophobic Block of Pluronics in Cells: a Photo Affinity Labelling Approach. Pharm Res 35, 205 (2018). https://doi.org/10.1007/s11095-018-2484-4

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