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AAPS PharmSciTech

, Volume 19, Issue 1, pp 201–212 | Cite as

Antiproliferative Activity and VEGF Expression Reduction in MCF7 and PC-3 Cancer Cells by Paclitaxel and Imatinib Co-encapsulation in Folate-Targeted Liposomes

  • Marco Júnio Peres-Filho
  • Alexandre Pereira dos Santos
  • Thais Leite Nascimento
  • Renato Ivan de Ávila
  • Fabrícia Saba Ferreira
  • Marize Campos Valadares
  • Eliana Martins LimaEmail author
Research Article
  • 436 Downloads

Abstract

Co-encapsulation of anticancer drugs paclitaxel and imatinib in nanocarriers is a promising strategy to optimize cancer treatment. Aiming to combine the cytotoxic and antiangiogenic properties of the drugs, a liposome formulation targeted to folate receptor co-encapsulating paclitaxel and imatinib was designed in this work. An efficient method was optimized for the synthesis of the lipid anchor DSPE-PEG(2000)-folic acid (FA). The structure of the obtained product was confirmed by RMN, FT-IR, and ESI-MS techniques. A new analytical method was developed and validated for simultaneous quantification of the drugs by liquid chromatography. Liposomes, composed of phosphatidylcholine, cholesterol, and DSPE-mPEG(2000), were prepared by extrusion. Their surface was modified by post-insertion of DSPE-PEG(2000)-FA. Reaction yield for DSPE-PEG(2000)-FA synthesis was 87%. Liposomes had a mean diameter of 122.85 ± 1.48 nm and polydispersity index of 0.19 ± 0.01. Lyophilized formulations remained stable for 60 days in terms of size and drug loading. FA-targeted liposomes had a higher effect on MCF7 cell viability reduction (p < 0.05) when compared with non-targeted liposomes and free paclitaxel. On PC-3 cells, viability reduction was greater (p < 0.01) when cells were exposed to targeted vesicles co-encapsulating both drugs, compared with the non-targeted formulation. VEGF gene expression was reduced in MCF7 and PC-3 cells (p < 0.0001), with targeted vesicles exhibiting better performance than non-targeted liposomes. Our results demonstrate that multifunctional liposomes associating molecular targeting and multidrug co-encapsulation are an interesting strategy to achieve enhanced internalization and accumulation of drugs in targeted cells, combining multiple antitumor strategies.

KEY WORDS

liposomes co-encapsulation folate receptor tumor targeting VEGF 

Notes

Acknowledgments

This work was financially supported by the following Brazilian research funding agencies: Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Financiadora de Estudos e Pesquisas (FINEP), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Apoio à Pesquisa da Universidade Federal de Goiás (FUNAPE), and Fundação de Apoio à Pesquisa do Estado de Goiás (FAPEG). The authors would like to thank Prof. Dr. Cecília Maria Alves de Oliveira and Dr. Monique Ferreira Marques from the Chemistry Institute/Federal University of Goias, for the assistance with synthesis and chemical characterization of the phospholipid anchor.

Author Contributions

Not applicable.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflicts of interest.

Supplementary material

12249_2017_830_Fig8_ESM.gif (19 kb)
Fig. S1

a 1H NMR spectrum of the DSPE-PEG(2000)-FA reaction product and b detail of the spectrum. Notes: The spectra obtained demonstrated regions containing aliphatic hydrogens at δ 0.87 (t, CH3, 6H), δ 1.25 (s, CH2, ~57H), δ 1.58 (m, CH2CH2CO, 4H), δ 2.27 (m, CH2, ~6H), δ 3.96 (m, CH2CH2N, 4H), and δ 3.66 (s, PEG, ~175H). Additionally, signals related to hydrogen next to the amide and phosphate groups were observed at δ 4.18 (t, CH2OCONH, 2H), δ 4.37 (dd, trans-PO4CH2CH, 1H), and δ 5.21 (m, PO4CH2CH, 1H), respectively, indicating binding of DSPE-PEG(2000)-NH2 to folic acid. The hydrogen related to the aromatic moiety of the molecule were observed at δ 6.62 (m, 3′,5′-H, 1H) and δ 7.91 (m, 1H) (Fig. S1 B). Abbreviations: DSPE-PEG(2000)-FA 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(folate(polyethylene glycol)-2000), DSPE-PEG-NH 2 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(amino(polyethylene glycol)-2000). (GIF 19 kb)

12249_2017_830_MOESM1_ESM.tif (260 kb)
High-resolution image (TIFF 260 kb)

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Copyright information

© American Association of Pharmaceutical Scientists 2017

Authors and Affiliations

  • Marco Júnio Peres-Filho
    • 1
    • 2
  • Alexandre Pereira dos Santos
    • 3
  • Thais Leite Nascimento
    • 2
  • Renato Ivan de Ávila
    • 3
  • Fabrícia Saba Ferreira
    • 2
  • Marize Campos Valadares
    • 3
  • Eliana Martins Lima
    • 2
    • 4
    Email author
  1. 1.State University of GoiásCampus ItumbiaraBrazil
  2. 2.Laboratory of Pharmaceutical Nanotechnology and Drug Delivery Systems, School of PharmacyFederal University of GoiásGoiâniaBrazil
  3. 3.Laboratory of Cellular Pharmacology and Toxicology, School of PharmacyFederal University of GoiásGoiâniaBrazil
  4. 4.Faculdade de FarmáciaUniversidade Federal de GoiásGoiâniaBrazil

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