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Pharmaceutical Research

, 35:77 | Cite as

A Smart Paclitaxel-Disulfiram Nanococrystals for Efficient MDR Reversal and Enhanced Apoptosis

  • Imran Shair Mohammad
  • Wei He
  • Lifang Yin
Research Paper

Abstract

Purpose

A multidrug resistance (MDR) modulator, disulfiram (DSF), was incorporated into pure paclitaxel (PTX) nanoparticles to construct a smart paclitaxel-disulfiram nanococrystals (PTX-DSF Ns) stabilized by β-lactoglobulin (β-LG), with the aim to reverse MDR and therefore enhnce cytotoxicity towards Taxol-resistant A549 cells (A549/TAX).

Method

PTX-DSF Ns was prepared by antisolvent precipitation method. Flow cytometry was used to determine the cell uptake, drug efflux inhibition, cell cycle phase arrest and apoptosis. MDR-1 gene expression level was detected by real time quantitative PCR and gel electrophoresis.

Results

PTX-DSF Ns prepared from the optimized formulation had an optimum diameter of 160 nm, was stable and had a high drug-loading capacity. Importantly, the uptake of PTX-DSF Ns in A549/TAX cells was 14-fold greater than the uptake of PTX Ns. Furthermore, PTX-DSF Ns promoted 5-folds increase in apoptosis, enabled 7-folds reduction in the IC50, and rendered 8.9-fold decrease in the dose compared with free PTX.

Conclusion

PTX-DSF Ns with a precise mass ratio offer efficient cytotoxicity against Taxol-resistant cells and a novel approach for codelivery and sensitizing MDR cancer to chemotherapy. In addition, the use of nanosuspensions as a combined treatment provides a new research avenue for nanosuspensions.

Keywords

apoptosis combined therapy multidrug resistance nanosuspensions p-glycoprotein 

Abbreviations

A549

Sensitive human lung adenocarcinoma cell line

A549/TAX

Taxol resistant human lung adenocarcinoma cell line

ABC

ATP-Binding Cassette

BCA

Bicinchoninic acid

β-LG

β-lactoglobulin

CD

Circular Dichroism

cDNA

Complementary DNA

CI

Combination index

CLSM

Confocal laser scanning microscope

DAPI

4, 6-diamidino-2-phenylindole

DL

Drug loading

DLS

Dynamic light scattering

DMSO

Dimethyl sulfoxide

DRI

Dose reduction index

DSF

Disulfiram

EE

Encapsulation efficiency

FBS

Fetal bovine serum

FDA

Food and Drug administration

FITC

Fluorescein 5(6)-isothiocyanate

HPLC

High-performance liquid chromatography

IC50

Half maximal inhibitory concentration

MDR

Multidrug resistance

MDR-1

Multidrug resistance gene-1

MTT

3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide

PBS

Phosphate buffer saline

PDI

Polydispersity index

P-gp

P-glycoprotein

PI

Propidium iodide

PTX

Paclitaxel

PTX-DSF

Free Paclitaxel-Disulfiram formulation

PTX-DSF Ns

Paclitaxel-Disulfiram nanococrystals

PXRD

Powder X-ray Diffraction

RNA

Ribonucleic acid

RT-qPCR

Real time quantitative PCR

SEM

Scanning electron microscopy

TEM

Transmission electron microscopy

Trp

Tryptophan

Notes

Acknowledgments and Disclosures

This study was supported by grants from the National Natural Science Foundation of China (Nos. 81673377, 81473152, and 81402869), the Natural Science Foundation of Jiangsu Province (No. BK20140671), and the Fostering Plan of University Scientific and Technological Innovation Team and Key Members of the Outstanding Young Teacher of Jiangsu Qing Lan Project (2014 and 2016). We also thank Xiaonan Ma, Minhui Sun and Yingjian Hou from the Cellular and Molecular Biology Center of China Pharmaceutical University for providing technical assistance. The authors report no conflicts of interest with this work.

Supplementary material

11095_2018_2370_MOESM1_ESM.docx (1.3 mb)
ESM 1 (DOCX 1281 kb)

References

  1. 1.
    Hirsch FR, Suda K, Wiens J, Bunn PA Jr. New and emerging targeted treatments in advanced non-small-cell lung cancer. Lancet. 2016;388:1012–24.CrossRefPubMedGoogle Scholar
  2. 2.
    Hirsch FR, Scagliotti GV, Mulshine JL, Kwon R, Curran WJ, Wu YL, et al. Lung cancer: current therapies and new targeted treatments. Lancet. 2017;389:299–311.CrossRefPubMedGoogle Scholar
  3. 3.
    Saad M, Garbuzenko OB, Minko T. Co-delivery of siRNA and an anticancer drug for treatment of multidrug-resistant cancer. Nanomedicine (London). 2008;3:761–76.CrossRefGoogle Scholar
  4. 4.
    Gao Z, Zhang L, Sun Y. Nanotechnology applied to overcome tumor drug resistance. J Control Release. 2012;162:45–55.CrossRefPubMedGoogle Scholar
  5. 5.
    Li W, Zhang H, Assaraf YG, Zhao K, Xu X, Xie J, et al. Overcoming ABC transporter-mediated multidrug resistance: molecular mechanisms and novel therapeutic drug strategies. Drug Resist Updat. 2016;27:14–29.CrossRefPubMedGoogle Scholar
  6. 6.
    Sandberg T, Rosenholm J, Hotokka M. The molecular structure of disulfiram and its complexation with silica. A quantum chemical study. J Mol Struc-Theochem. 2008;861:57–61.CrossRefGoogle Scholar
  7. 7.
    Sauna ZE, Peng XH, Nandigama K, Tekle S, Ambudkar SV. The molecular basis of the action of disulfiram as a modulator of the multidrug resistance-linked ATP binding cassette transporters MDR1 (ABCB1) and MRP1 (ABCC1). Mol Pharmacol. 2004;65:675–84.CrossRefPubMedGoogle Scholar
  8. 8.
    Duan X, Xiao J, Yin Q, Zhang Z, Yu H, Mao S, et al. Smart pH-sensitive and temporal-controlled polymeric micelles for effective combination therapy of doxorubicin and disulfiram. ACS Nano. 2013;7:5858–69.CrossRefPubMedGoogle Scholar
  9. 9.
    Lehar J, Krueger AS, Avery W, Heilbut AM, Johansen LM, Price ER, et al. Synergistic drug combinations tend to improve therapeutically relevant selectivity. Nat Biotechnol. 2009;27:659–66.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Hu CMJ, Aryal S, Zhang L. Nanoparticle-assisted combination therapies for effective cancer treatment. Ther Deliv. 2010;1:323–34.CrossRefPubMedGoogle Scholar
  11. 11.
    Mehra NK, Jain K, Jain NK. Pharmaceutical and biomedical applications of surface engineered carbon nanotubes. Drug Discov Today. 2015;20:750–9.CrossRefPubMedGoogle Scholar
  12. 12.
    Misra R, Acharya S, Sahoo SK. Cancer nanotechnology: application of nanotechnology in cancer therapy. Drug Discov Today. 2010;15:842–50.CrossRefPubMedGoogle Scholar
  13. 13.
    Gao H. Progress and perspectives on targeting nanoparticles for brain drug delivery. Acta Pharm Sin B. 2016;6:268–86.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Shen Y, Jin E, Zhang B, Murphy CJ, Sui M, Zhao J, et al. Prodrugs forming high drug loading multifunctional nanocapsules for intracellular cancer drug delivery. J Am Chem Soc. 2010;132:4259–65.CrossRefPubMedGoogle Scholar
  15. 15.
    Guo S, Lin CM, Xu Z, Miao L, Wang Y, Huang L. Co-delivery of cisplatin and rapamycin for enhanced anticancer therapy through synergistic effects and microenvironment modulation. ACS Nano. 2014;8:4996–5009.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Rabinow BE. Nanosuspensions in drug delivery. Nat Rev Drug Discov. 2004;3:785–96.CrossRefPubMedGoogle Scholar
  17. 17.
    He W, Xin X, Li Y, Han X, Qin C, Yin L. Rod-shaped drug particles for cancer therapy: the importance of particle size and participation of caveolae pathway. Part Part Syst Charact. 2017;34(6)Google Scholar
  18. 18.
    He W, Wang Y, Lv Y, Xiao Q, Ye L, Cai B, et al. Denatured protein stabilized drug nanoparticles: tunable drug state and penetration across the intestinal barrier. J Mater Chem B. 2017;5:1081–97.CrossRefGoogle Scholar
  19. 19.
    Attili-Qadri S, Karra N, Nemirovski A, Schwob O, Talmon Y, Nassar T, et al. Oral delivery system prolongs blood circulation of docetaxel nanocapsules via lymphatic absorption. P Natl Acad Sci USA. 2013;110:17498–503.CrossRefGoogle Scholar
  20. 20.
    Fuhrmann K, Gauthier MA, Leroux JC. Targeting of injectable drug nanocrystals. Mol Pharm. 2014;11:1762–71.CrossRefPubMedGoogle Scholar
  21. 21.
    Müller RH, Gohla S, Keck CM. State of the art of nanocrystals – special features, production, nanotoxicology aspects and intracellular delivery. Eur J Pharm Biopharm. 2011;78:1–9.CrossRefPubMedGoogle Scholar
  22. 22.
    Lu Y, Qi J, Dong X, Zhao W, Wu W. The in vivo fate of nanocrystals. Drug Discov Today. 2017;22:744–50.CrossRefPubMedGoogle Scholar
  23. 23.
    Gao H. Perspectives on dual targeting delivery systems for brain tumors. J Neuroimmune Pharmacol. 2017;12:6–16.CrossRefPubMedGoogle Scholar
  24. 24.
    Li Y, Wu Z, He W, Qin C, Yao J, Zhou J, et al. Globular protein-coated paclitaxel nanosuspensions:interaction mechanism, direct cytosolic delivery, and significant improvement in pharmacokinetics. Mol Pharm. 2015;12:1485–500.CrossRefPubMedGoogle Scholar
  25. 25.
    He W, Lu Y, Qi J, Chen L, Hu F, Wu W. Food proteins as novel nanosuspension stabilizers for poorly water-soluble drugs. Int J Pharm. 2013;441:269–78.CrossRefPubMedGoogle Scholar
  26. 26.
    Zhao R, Hollis CP, Zhang H, Sun L, Gemeinhart RA, Li T. Hybrid nanocrystals: achieving concurrent therapeutic and bioimaging functionalities toward solid tumors. Mol Pharm. 2011;8:1985–91.CrossRefPubMedGoogle Scholar
  27. 27.
    Chen SY, Hu SS, Dong Q, Cai JX, Zhang WP, Sun JY, et al. Dong. Establishment of paclitaxel-resistant breast cancer cell line and nude mice models, and underlying multidrug resistance mechanisms in vitro and in vivo. Asian Pac J Cancer Prev. 2013;14:6135–40.CrossRefPubMedGoogle Scholar
  28. 28.
    Jiang N, Dong XP, Zhang SL, You QY, Jiang XT, Zhao XG. Triptolide reverses the taxol resistance of lung adenocarcinoma by inhibiting the NF-κB signaling pathway and the expression of NF-κB-regulated drug-resistant genes. Mol Med Rep. 2016;13:153–9.CrossRefPubMedGoogle Scholar
  29. 29.
    Chou T-C. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev. 2006;58:621–81.CrossRefPubMedGoogle Scholar
  30. 30.
    Tallarida RJ. An overview of drug combination analysis with isobolograms. J Pharmacol Exp Ther. 2006;319:1–7.CrossRefPubMedGoogle Scholar
  31. 31.
    Li J, Yang YL, Li LZ, Zhang L, Liu Q, Liu K, et al. Succinate accumulation impairs cardiac pyruvate dehydrogenase activity through GRP91-dependent and independent signaling pathways: therapeutic effects of ginsenoside Rb1. Biochim Biophys Acta. 2017;1863(11):2835–47.CrossRefPubMedGoogle Scholar
  32. 32.
    Ha HK, Kim JW, Lee MR, Jun W, Lee WJ. Cellular uptake and cytotoxicity of β-lactoglobulin nanoparticles: the effects of particle size and surface charge. Asian Australas J Anim Sci. 2015;28:420–7.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Song W, Tang Z, Lei T, Wen X, Wang G, Zhang D, et al. Stable loading and delivery of disulfiram with mPEG-PLGA/PCL mixed nanoparticles for tumor therapy. Nanomed-Nanotechnol. 2016;12:377–86.CrossRefGoogle Scholar
  34. 34.
    Duan X, Li Y. Physicochemical characteristics of nanoparticles affect circulation, biodistribution, cellular internalization, and trafficking. Small. 2013;9:1521–32.CrossRefPubMedGoogle Scholar
  35. 35.
    Shen XC, Liou XY, Ye LP, Liang H, Wang ZY. Spectroscopic studies on the interaction between human hemoglobin and CdS quantum dots. J Colloid Interf Sci. 2007;311:400–6.CrossRefGoogle Scholar
  36. 36.
    Gorinstein S, Goshev I, Moncheva S, Zemser M, Weisz M, Caspi A, et al. Intrinsic tryptophan fluorescence of human serum proteins and related conformational changes. J Protein Chem. 2000;19:637–42.CrossRefPubMedGoogle Scholar
  37. 37.
    Claytonand AH, Sawyer WH. Site-specific tryptophan fluorescence spectroscopy as a probe of membrane peptide structure and dynamics. Eur Biophys J Biophys. 2002;31:9–13.CrossRefGoogle Scholar
  38. 38.
    Chadborn N, Bryant J, Bain AJ, Shea PO. Ligand-dependent conformational equilibria of serum albumin revealed by tryptophan fluorescence quenching. Biophys J. 1999;76:2198–207.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Roach P, Farrar D, Perry CC. Interpretation of protein adsorption: surface-induced conformational changes. J Am Chem Soc. 2005;127:8168–73.CrossRefPubMedGoogle Scholar
  40. 40.
    Brewer SH, Glomm WR, Johnson MC, Knag MK, Franzen S. Probing BSA binding to citrate-coated gold nanoparticles and surfaces. Langmuir. 2005;21:9303–7.CrossRefPubMedGoogle Scholar
  41. 41.
    Gao D, Tian Y, Bi S, Chen Y, Yu A, Zhang H. Studies on the interaction of colloidal gold and serum albumins by spectral methods. Spectrochim Acta A. 2005;62:1203–8.CrossRefGoogle Scholar
  42. 42.
    Xiao Q, Huang S, Qi ZD, Zhou B, He ZK, Liu Y. Conformation, thermodynamics and stoichiometry of HSA adsorbed to colloidal CdSe/ZnS quantum dots. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 2008;1784:1020–7.CrossRefGoogle Scholar
  43. 43.
    Jiang L, Yang BQ, Ma YD, Liu YC, Yang WS, Li TJ, et al. The binding of phosphorothioate oligonucleotides to CdS nanoparticles. Chem Phys Lett. 2003;380:29–33.CrossRefGoogle Scholar
  44. 44.
    Kelly SM, Jess TJ, Price NC. How to study proteins by circular dichroism. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 2005;1751:119–39.CrossRefGoogle Scholar
  45. 45.
    Zhang L, Xiao Q, Wang Y, Zhang C, He W, Yin L. Denatured protein-coated docetaxel nanoparticles: alterable drug state and cytosolic delivery. Int J Pharm. 2017;523:1–14.CrossRefPubMedGoogle Scholar
  46. 46.
    Loo TW, Clarke DM. Blockage of drug resistance in vitro by disulfiram, a drug used to treat alcoholism. J Natl Cancer Inst. 2000;92:898–902.CrossRefPubMedGoogle Scholar
  47. 47.
    Fasehee H, Dinarvand R, Ghavamzadeh A, Esfandyari-Manesh M, Moradian H, Faghihi S, et al. Delivery of disulfiram into breast cancer cells using folate-receptor-targeted PLGA-PEG nanoparticles: in vitro and in vivo investigations. J Nanobiotecnol. 2016;14:32.CrossRefGoogle Scholar
  48. 48.
    Cintron-Colonand R, Vega I. Molecular mechanism of paclitaxel-induced degradation of SCG10: a potential neuropathy biomarker. FASEB J. 2014;28:651–14.Google Scholar
  49. 49.
    Saw PE, Park J, Jon S, Farokhzad OC. A drug-delivery strategy for overcoming drug resistance in breast cancer through targeting of oncofetal fibronectin. Nanomed-Nanotechnol. 2017;13:713–22.CrossRefGoogle Scholar
  50. 50.
    Gao L, Liu G, Ma J, Wang X, Zhou L, Li X. Drug nanocrystals: in vivo performances. J Control Release. 2012;160:418–30.CrossRefPubMedGoogle Scholar
  51. 51.
    Chen Y, Li T. Cellular uptake mechanism of paclitaxel nanocrystals determined by confocal imaging and kinetic measurement. AAPS J. 2015;17:1126–34.CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Fuhrmann K, Polomska A, Aeberli C, Castagner B, Gauthier MA, Leroux JC. Modular design of redox-responsive stabilizers for nanocrystals. ACS Nano. 2013;7:8243–50.CrossRefPubMedGoogle Scholar
  53. 53.
    Hu Q, Sun W, Wang C, Gu Z. Recent advances of cocktail chemotherapy by combination drug delivery systems. Adv Drug Deliv Rev. 2016;98:19–34.CrossRefPubMedGoogle Scholar
  54. 54.
    Noh J, Kwon B, Han E, Park M, Yang W, Cho W, et al. Amplification of oxidative stress by a dual stimuli-responsive hybrid drug enhances cancer cell death. Nat Commun. 2015;6:6907.CrossRefPubMedGoogle Scholar
  55. 55.
    Chen YZ, Zhang M, Jin HY, Tang YS, Wu AH, Huang YZ. Prodrug-like, PEGylated protein toxin trichosanthin for reversal of chemoresistance. Mol Pharm. 2017;14:1429–38.CrossRefPubMedGoogle Scholar
  56. 56.
    Ni J, Tian FC, Dahmani FZ, Yang H, Yue DR, He SW, et al. Curcumin-carboxymethyl chitosan (CNC) conjugate and CNC/LHR mixed polymeric micelles as new approaches to improve the oral absorption of P-gp substrate drugs. Drug Deliv. 2016;23:3424–35.CrossRefPubMedGoogle Scholar
  57. 57.
    Baird RD, Tan DS, Kaye SB. Weekly paclitaxel in the treatment of recurrent ovarian cancer. Nat Rev Clin Oncol. 2010;7:575–82.CrossRefPubMedGoogle Scholar
  58. 58.
    Liu Y, Huang L, Liu F. Paclitaxel nanocrystals for overcoming multidrug resistance in cancer. Mol Pharm. 2010;7:863–9.CrossRefPubMedPubMedCentralGoogle Scholar
  59. 59.
    Johansson B. Stabilization and quantitative determination of disulfiram in human plasma samples. Clin Chim Acta. 1988;177:55–63.CrossRefPubMedGoogle Scholar
  60. 60.
    B. Johansson. A review of the pharmacokinetics and pharmacodynamics of disulfiram and its metabolites. Acta psychiatrica Scandinavica Supplementum. 1992;369:15–26.Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Pharmaceutics, School of PharmacyChina Pharmaceutical UniversityNanjingPeople’s Republic of China
  2. 2.Key Laboratory of Druggability of BiopharmaceuticsChina Pharmaceutical UniversityNanjingPeople’s Republic of China

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