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. 2019 Apr 10;14:2573-2589.
doi: 10.2147/IJN.S195934. eCollection 2019.

Characterization and Bioactivity of Self-Assembled Anti-Angiogenic Chondroitin sulfate-ES2-AF Nanoparticle Conjugate

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Free PMC article

Characterization and Bioactivity of Self-Assembled Anti-Angiogenic Chondroitin sulfate-ES2-AF Nanoparticle Conjugate

Liang Xing et al. Int J Nanomedicine. .
Free PMC article

Abstract

Background: In the past few years, significant progress has been made in inhibiting neovascularization at the tumor site, cutting off the nutrient supply of the tumor, and inhibiting tumor growth and metastasis. However, many proteins/peptides have the disadvantage of poor stability, short half-life, and uncertain targeting ability. Chemical modification can be used to overcome these disadvantages; many polyethylene glycol-modified proteins/peptides have been approved by US FDA. The purpose of this study was to obtain a novel anti-angiogenic chondroitin sulfate (CS)-peptide nanoparticle conjugate with efficient anti-neovascularization and tumor targeting ability and an acceptable half-life.

Materials and methods: The CS-ES2-AF nanoparticle conjugate was synthesized and characterized using 1H-nuclear magnetic resonance spectroscopy, transmission electron microscopy, and particle size and zeta potential analyzer. The anti-angiogenic ability was studied using MTT, migration, tube formation, and chick chorioallantoic membrane assays. The targeting ability of CS-ES2-AF was studied by ELISA, surface plasmon resonance, and bioimaging. The pharmacokinetics was also studied.

Results: The CS-ES2-AF could self-assemble into stable nanoparticles in aqueous solution, which significantly enhances its anti-neovascularization activity, tumor targeting more explicit, and prolongs its half-life.

Conclusion: CS is an effective protein/peptide modifier, and CS-ES2-AF displayed good potential in tumor targeting therapy.

Keywords: ES2-AF; anti-angiogenesis; chondroitin sulfate; nanoparticles; targeting.

Conflict of interest statement

Disclosure The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
Synthesis and 1H-NMR spectra of CS-ES2-AF. (A) Synthetic route of CS-ES2-AF. 1H-NMR spectra of (B) CS, (C) CS-TBA, and (D) CS-ES2-AF. Abbreviations: CS, chondroitin sulfate; 1H-NMR, 1H-nuclear magnetic resonance spectroscopy.
Figure 2
Figure 2
Characterization of CS-ES2-AF nanoparticles. (A) TEM image of CS-ES2-AF and ES2-AF nanoparticles. (B) Size distribution of CS-ES2-AF and ES2-AF nanoparticles. Abbreviations: CS, chondroitin sulfate; TEM, transmission electron microscopy.
Figure 3
Figure 3
(A) Inhibitory effects of peptide ES2-AF and conjugate CS-ES2-AF on the proliferation of EA.hy926 endothelial cells after incubation for 48 hours. (B) Comparison of the effects of ES2-AF, the mixture of CS&ES2-AF, and CS-ES2-AF conjugation based on in vitro ELISA assay. Data represent mean ± SD (n=5). **P<0.01 is considered significantly different compared with the ES2-AF group. Abbreviation: CS, chondroitin sulfate.
Figure 4
Figure 4
Effects of different samples on the inhibition of cell migration, tube formation, and CAM: (i) represents the ES2-AF sample; (ii) represents the CS-ES2-AF sample; and (iii) represents the CS&ES2-AF sample. (A) Cell migration assay. (B) Tube formation assay. (C) CAM assay. Comparison of different samples for (D) cell migration and (E) tube formation. (F) Comparison of the inhibition of ES2-AF and its derivatives and the mixture on CAM angiogenesis. Data represent mean ± SD (n=5). #P<0.05 is considered significantly different compared with the control group. ##P<0.01 is considered significantly different compared with the control group. *P<0.05 is considered significantly different compared with the ES2-AF group. **P<0.01 is considered significantly different compared with the ES2-AF group.
Figure 4
Figure 4
Effects of different samples on the inhibition of cell migration, tube formation, and CAM: (i) represents the ES2-AF sample; (ii) represents the CS-ES2-AF sample; and (iii) represents the CS&ES2-AF sample. (A) Cell migration assay. (B) Tube formation assay. (C) CAM assay. Comparison of different samples for (D) cell migration and (E) tube formation. (F) Comparison of the inhibition of ES2-AF and its derivatives and the mixture on CAM angiogenesis. Data represent mean ± SD (n=5). #P<0.05 is considered significantly different compared with the control group. ##P<0.01 is considered significantly different compared with the control group. *P<0.05 is considered significantly different compared with the ES2-AF group. **P<0.01 is considered significantly different compared with the ES2-AF group.
Figure 5
Figure 5
Binding ability of CS-ES2-AF to the CD44 receptor in vitro. (A) pH scouting of CD44 in acetate buffer solution. (B) SPR sensorgram of different concentrations of CS-ES2-AF over a chip with immobilized CD44. (C) SPR sensorgram of different concentrations of ES2-AF over a chip with immobilized CD44. Abbreviations: CS, chondroitin sulfate; SPR, surface plasmon resonance.
Figure 6
Figure 6
In vivo tumor-targeting ability of CS-ES2-AF in nude mice bearing B16 tumor xenografts, monitored by the Optix system. Tumor is indicated using red circle. Abbreviation: CS, chondroitin sulfate.
Figure 7
Figure 7
Pharmacokinetic time curves of ES2-AF and CS-ES2-AF in plasma after single intravenous administration. The results are shown as means ± SD (n=5). Abbreviation: CS, chondroitin sulfate.

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References

    1. Novoa-Carballal R, Perez-Martin R, Blanco M, et al. By-products of Scyliorhinus canicula, Prionace glauca and Raja clavata: A valuable source of predominantly 6S sulfated chondroitin sulfate. Carbohydr Polym. 2017;157:31–37. doi: 10.1016/j.carbpol.2016.09.050. - DOI - PubMed
    1. Yamada S, Sugahara K. Potential therapeutic application of chondroitin sulfate/dermatan sulfate. Curr Drug Discov Technol. 2008;5(4):289–301. - PubMed
    1. Zhang JS, Imai T, Suenaga A, Otagiri M. Molecular-weight-dependent pharmacokinetics and cytotoxic properties of cisplatin complexes prepared with chondroitin sulfate A and C. Int J Pharm. 2002;240(1–2):23–31. - PubMed
    1. Onishi H, Matsuyama M. Conjugate between chondroitin sulfate and prednisolone with a glycine linker: preparation and in vitro conversion analysis. Chem Pharm Bull (Tokyo) 2013;61(9):902–912. - PubMed
    1. Lo YL, Sung KH, Chiu CC, Wang LF. Chemically conjugating polyethylenimine with chondroitin sulfate to promote CD44-mediated endocytosis for gene delivery. Mol Pharm. 2013;10(2):664–676. doi: 10.1021/mp300432s. - DOI - PubMed

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