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SCIENCE CHINA Materials, Volume 61, Issue 11: 1462-1474(2018) https://doi.org/10.1007/s40843-018-9277-8

Enzyme/pH-sensitive dendritic polymer-DOX conjugate for cancer treatment

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  • ReceivedFeb 20, 2018
  • AcceptedApr 10, 2018
  • PublishedMay 22, 2018

Abstract

It is in a great demand to design a biodegradable, tumor microenvironment-sensitive drug delivery system to achieve safe and highly efficacious treatment of cancer. Herein, a novel pH/enzyme sensitive dendritic pdiHPMA-DOX conjugate was designed. diHPMA dendritic copolymer with GFLG segments in the branches which are sensitive to the intracellular enzyme of the tumor was prepared through RAFT polymerization. DOX was attached to dendritic diHPMA polymer through a pH-sensitive hydrazone bond. The dendritic pdiHPMA-DOX conjugate self-assembled into nanoparticles with an ideal spherical shape at a mean size of 103 nm. The DOX attached to the polymeric carrier was released in an acidic environment, and the GFLG linker for synthesizing the dendritic vehicle with a high molecular weight (MW, 220 kDa) was cleaved to release low MW segments (<40 kDa) in the presence of cathepsin B. The dendritic polymeric conjugate was internalized via an endocytic pathway, and then released the anticancer drug, which led to significant cytotoxicity for tumors. The blood circulation time was profoundly prolonged, resulting in high accumulation of DOX into tumors. In vivo anti-tumor experiments with 4T1 tumor bearing mice demonstrated that the conjugate had a better antitumor efficacy in comparison with free DOX. Additionally, body weight measurements and histological examinations indicated that the conjugate showed low toxicities to normal tissues. This dendritic polymeric drug carrier in a response to intracellular enzyme and acidic pH of tumor tissue or cells holds great promise in tumor-targeted therapy.


Funded by

the National Natural Science Foundation of China(51673127,8162103)

International Science and Technology Cooperation Program of China(2015DFE52780,81220108013)

International Science and Technology Cooperation Program of Chengdu(2016-GH03-00005-HZ)


Acknowledgment

This work was supported by the National Natural Science Foundation of China (51673127 and 8162103), International Science and Technology Cooperation Program of China (2015DFE52780 and 81220108013) and International Science and Technology Cooperation Program of Chengdu (2016-GH03-00005-HZ).


Interest statement

The authors declare that they have no conflict of interest.


Contributions statement

Chen K designed the study, performed the experiments, analyzed the data, and wrote the manuscript. Liao S and Guo S performed experiments. Zhang H and Cai H analyzed the data, wrote the manuscript. Gong Q, Gu Z and Luo K designed the study, wrote the manuscript and they are responsible for funding support, resources and project administration.


Author information

Kai Chen received his BSc degree from Army Medical University (Third Military Medical University), Chongqing, in 2015. Now he is a PhD candidate at the National Engineering Research Center for Biomaterials, Sichuan University. His current research focuses on the treatment of breast cancer with enzyme/pH sensitive polymer-drug conjugates-based nanoscale delivery system.


Kui Luo is a Professor in West China Hospital and National Engineering Research Center for Biomaterials, Sichuan University, China. He obtained his PhD degree from the National Engineering Research Center for Biomaterials, Sichuan University in 2009, and then became an assistant professor in this center. From 2009 to 2011, he carried out his postdoctoral work on polymeric nanomedicines at the Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, USA. Dr. Luo was promoted to an associate professor in 2012 and Full Professor in 2013 in Sichuan University. From 2016, he was also a Full professor in Huaxi MR Research Center (HMRRC), Department of Radiology, West China Hospital, Sichuan University. His research focuses on stimuli-responsive and biodegradable polymeric gene/drug delivery vehicles and imaging probes for cancer diagnosis and therapy, especially the study of synthetic macromolecules as potential cancer therapeutic and diagnostic agents, and the relationships between their actions and structural features.


Supplement

Supplementary information

Supporting data are available in the online version of the paper.


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  • Figure 1

    The 1H NMR spectra of the dendritic pdiHPMA-DOX conjugate recorded in D2O.

  • Scheme 1

    Preparation of the dendritic pdiHPMA-DOX conjugate.

  • Figure 2

    Particle size of dendritic conjugate. (a) SEM images. (b) Size distribution by dynamic light scattering (DLS).

  • Figure 3

    (a) SEC profiles of dendritic conjugate and degraded product. (b) Cumulative DOX release profile from the dendritic conjugate at pH 5.0 and pH 7.4 at 37°C. The buffer was mixed with or without cathepsin B. The data shown are mean ± SD (n=3).

  • Figure 4

    Cytotoxicity of the dendritic conjugate (a) and the drug-free dendritic conjugate against 4T1 cells (b) incubation for 48 h at different concentrations. The data shown are mean ± SD (n=5).

  • Figure 5

    In vitro cellular uptake of free DOX (a) and the dendritic conjugate (b) in 4T1 cells after incubation for 0.5, 2, and 4 h under a CLSM. Cell nuclei were stained with Hoechst 33342. Bar = 25 μm.

  • Figure 6

    (a) Confocal images of cellular uptake of the dendritic conjugate by 4T1 cells after 4 h of incubation with the conjugate at 37°C. The acidic organelles were stained with Lysotracker Green, and cell nuclei with Hoechst 33342. Scale bars: 25 μm. (b) Analysis of 4T1 cell apoptosis induced by free DOX and the dendritic conjugate after 48 h incubation by flow cytometry.

  • Figure 7

    Pharmacokinetic profiles of free DOX and the dendritic conjugate after injection in healthy mice at a DOX dose of 5 mg kg−1 body weight. The data represent the mean ± SD (n=5).

  • Figure 8

    Anti-tumor study in the 4T1 breast tumor model (n=7). (a) The dendritic conjugate presented significant tumor suppression (**p<0.01, compared to saline; #p<0.05, compared to free DOX). The black arrows indicate the date of administration via tail vein. (b) On day 27, weights of tumor tissues. (**p<0.01, compared to saline; #p<0.05, compared to free DOX). (c) Tumor growth inhibition on the basis of the tumor weight. (d) Monitoring of body weight of the mice administrated with the dendritic conjugate, free DOX, and saline. (**p<0.01, free DOX in comparison with saline).

  • Figure 9

    H&E staining of major organs harvested from tumor-bearing mice administrated with saline, free DOX and the conjugate (×200).

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