The AAPS Journal

, 20:34 | Cite as

Targeted Delivery of Doxorubicin via CD147-Mediated ROS/pH Dual-Sensitive Nanomicelles for the Efficient Therapy of Hepatocellular Carcinoma

  • Chenxi Qu
  • Jizhao Li
  • Yejuan Zhou
  • Shudi Yang
  • Weiliang Chen
  • Fang Li
  • Bengang You
  • Yang Liu
  • Xuenong Zhang
Research Article


Low accumulation in tumor sites and slow intracellular drug release remain as the obstacles for nanoparticles to achieve effective delivery of chemotherapeutic drugs. In this study, multifunctional micelles were designed to deliver doxorubicin (Dox) to tumor sites to provide more efficient therapy against hepatic carcinoma. The micelles were based on pH-responsive carboxymethyl chitosan (CMCh) modified with a reactive oxygen species (ROS)-responsive segment phenylboronic acid pinacol ester (BAPE) and an active targeted ligand CD147 monoclonal antibody. The Dox-loaded micelles provided rapid and complete drug release in pH 5.3 incubation conditions with 1 mM H2O2. In addition, an in vitro cell uptake study revealed that CD147 modification significantly enhanced cellular internalization due to the high affinity to CD147 receptors, which are overexpressed on tumor cells. An in vivo study revealed that CD147-modified micellar formulations exhibited high accumulation in tumor sites and markedly enhanced antiproliferation effects with fewer side effects than other formulations. In conclusion, this CD147 receptor targeted delivery system with ROS/pH dual sensitivity provides a promising strategy for the treatment of hepatic carcinoma.


CD147 doxorubicin micelles ROS/pH dual sensitivity 



This work was supported by the National Natural Science Foundation of China (Nos. 81773183 and 81571788), Jiangsu Province Science and Technology Support Plan (BE2011670), and Priority Academic Program Development of Jiangsu Higher Education Institutions.


  1. 1.
    Maya S, Kumar LG, Sarmento B, Sanoj RN, Menon D, Nair SV, et al. Cetuximab conjugated O-carboxymethyl chitosan nanoparticles for targeting EGFR overexpressing cancer cells. Carbohydr Polym. 2013;93(2):661–9. Scholar
  2. 2.
    Kim DW, Kim SY, Kim HK, Kim SW, Shin SW, Kim JS, et al. Multicenter phase II trial of Genexol-PM, a novel Cremophor-free, polymeric micelle formulation of paclitaxel, with cisplatin in patients with advanced non-small-cell lung cancer. Ann Oncol. 2007;18(12):2009–14. Scholar
  3. 3.
    Nam JP, Park SC, Kim TH, Jang JY, Choi C, Jang MK, et al. Encapsulation of paclitaxel into lauric acid-O-carboxymethyl chitosan-transferrin micelles for hydrophobic drug delivery and site-specific targeted delivery. Int J Pharm. 2013;457(1):124–35. Scholar
  4. 4.
    Gong XY, Yin YH, Huang ZJ, Lu B, Xu PH, Zheng H, et al. Preparation, characterization and in vitro release study of a glutathione-dependent polymeric prodrug Cis-3-(9H-purin-6-ylthio)-acrylic acid-graft-carboxymethyl chitosan. Int J Pharm. 2012;436(1–2):240–7. Scholar
  5. 5.
    Prabhakar U, Maeda H, Jain RK, Sevick-Muraca EM, Zamboni W, Farokhzad OC, et al. Challenges and key considerations of the enhanced permeability and retention effect for nanomedicine drug delivery in oncology. Cancer Res. 2013;73(8):2412–7. Scholar
  6. 6.
    Moosavian SA, Abnous K, Badiee A, Jaafari MR. Improvement in the drug delivery and anti-tumor efficacy of PEGylated liposomal doxorubicin by targeting RNA aptamers in mice bearing breast tumor model. Colloids Surf, B. 2016;139:228–36. Scholar
  7. 7.
    Li C, Li H, Wang Q, Zhou M, Li M, Gong T, et al. pH-sensitive polymeric micelles for targeted delivery to inflamed joints. J Control Release. 2017;246:133–41. Scholar
  8. 8.
    Trachootham D, Alexandre J, Huang P. Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach? Nat Rev Drug Discov. 2009;8(7):579–91. Scholar
  9. 9.
    Khan M, Ong ZY, Wiradharma N, Attia AB, Yang YY. Advanced materials for co-delivery of drugs and genes in cancer therapy. Adv Healthc Mater. 2012;1(4):373–92. Scholar
  10. 10.
    Kim KS, Lee D, Song CG, Kang PM. Reactive oxygen species-activated nanomaterials as theranostic agents. Nanomedicine. 2015;10(17):2709–23. Scholar
  11. 11.
    Li N, Cai H, Jiang L, Hu J, Bains A, Hu J, et al. Enzyme-sensitive and amphiphilic PEGylated dendrimer-paclitaxel prodrug-pased nanoparticles for enhanced stability and anticancer efficacy. ACS Appl Mater Interfaces. 2017;9(8):6865–77. Scholar
  12. 12.
    Chen WL, Yang SD, Li F, Li JZ, Yuan ZQ, Zhu WJ, et al. Tumor microenvironment-responsive micelles for pinpointed intracellular release of doxorubicin and enhanced anti-cancer efficiency. Int J Pharm. 2016;511(2):728–40. Scholar
  13. 13.
    Singh B, Jiang T, Kim YK, Kang SK, Choi YJ, Cho CS. Release and cytokine production of bmpb from bmpb-loaded pH-sensitive and mucoadhesive thiolated eudragit microspheres. J Nanosci Nanotechnol. 2015;15(1):606–10. Scholar
  14. 14.
    Yuan F, Wang S, Chen G, Tu K, Jiang H, Wang LQ. Novel chitosan-based pH-sensitive and disintegrable polyelectrolyte nanogels. Colloids Surf, B. 2014;122:194–201. Scholar
  15. 15.
    Chen WL, Li F, Tang Y, Yang SD, Li JZ, Yuan ZQ, et al. Stepwise pH-responsive nanoparticles for enhanced cellular uptake and on-demand intracellular release of doxorubicin. Int J Nanomedicine. 2017;12:4241–56. Scholar
  16. 16.
    Liu X, Xiang J, Zhu D, Jiang L, Zhou Z, Tang J, et al. Fusogenic reactive oxygen species triggered charge-reversal vector for effective gene delivery. Adv Mater. 2016;28(9):1743–52. Scholar
  17. 17.
    Luo CQ, Xing L, Cui PF, Qiao JB, He YJ, Chen BA, et al. Curcumin-coordinated nanoparticles with improved stability for reactive oxygen species-responsive drug delivery in lung cancer therapy. Int J Nanomedicine. 2017;12:855–69. Scholar
  18. 18.
    Zhang D, Wei Y, Chen K, Zhang X, Xu X, Shi Q, et al. Biocompatible reactive oxygen species (ROS)-responsive nanoparticles as superior drug delivery vehicles. Adv Healthc Mater. 2015;4(1):69–76. Scholar
  19. 19.
    Wang F, Chen Y, Zhang D, Zhang Q, Zheng D, Hao L, et al. Folate-mediated targeted and intracellular delivery of paclitaxel using a novel deoxycholic acid-O-carboxymethylated chitosan-folic acid micelles. Int J Nanomedicine. 2012;7:325–37. Scholar
  20. 20.
    Guo H, Zhang D, Li C, Jia L, Liu G, Hao L, et al. Self-assembled nanoparticles based on galactosylated O-carboxymethyl chitosan-graft-stearic acid conjugates for delivery of doxorubicin. Int J Pharm. 2013;458(1):31–8. Scholar
  21. 21.
    Qi X, Qin J, Fan Y, Qin X, Jiang Y, Wu Z. Carboxymethyl chitosan-modified polyamidoamine dendrimer enables progressive drug targeting of tumors via pH-sensitive charge inversion. J Biomed Nanotechnol. 2016;12(4):667–78. Scholar
  22. 22.
    Kuang Y, Balakrishnan K, Gandhi V, Peng X. Hydrogen peroxide inducible DNA cross-linking agents: targeted anticancer prodrugs. J Am Chem Soc. 2011;133(48):19278–81. Scholar
  23. 23.
    Broaders KE, Grandhe S, Frechet JM. A biocompatible oxidation-triggered carrier polymer with potential in therapeutics. J Am Chem Soc. 2011;133(4):756–8. Scholar
  24. 24.
    Wang M, Sun S, Neufeld CI, Perez-Ramirez B, Xu Q. Reactive oxygen species-responsive protein modification and its intracellular delivery for targeted cancer therapy. Angew Chem Int Ed Engl. 2014;53(49):13444–8. Scholar
  25. 25.
    Naito M, Ishii T, Matsumoto A, Miyata K, Miyahara Y, Kataoka K. A phenylboronate-functionalized polyion complex micelle for ATP-triggered release of siRNA. Angew Chem Int Ed Engl. 2012;51(43):10751–5. Scholar
  26. 26.
    Zhu R, Zhang CG, Liu Y, Yuan ZQ, Chen WL, Yang SD, et al. CD147 monoclonal antibody mediated by chitosan nanoparticles loaded with alpha-hederin enhances antineoplastic activity and cellular uptake in liver cancer cells. Sci Rep. 2015;5(1):17904. Scholar
  27. 27.
    Zhu S, Qian L, Hong M, Zhang L, Pei Y, Jiang Y. RGD-modified PEG-PAMAM-DOX conjugate: in vitro and in vivo targeting to both tumor neovascular endothelial cells and tumor cells. Adv Mater. 2011;23(12):H84–9. Scholar
  28. 28.
    Wu FL, Zhang J, Li W, Bian BX, Hong YD, Song ZY, et al. Enhanced antiproliferative activity of antibody-functionalized polymeric nanoparticles for targeted delivery of anti-miR-21 to HER2 positive gastric cancer. Oncotarget. 2017;8(40):67189–202. Scholar
  29. 29.
    Kocbek P, Obermajer N, Cegnar M, Kos J, Kristl J. Targeting cancer cells using PLGA nanoparticles surface modified with monoclonal antibody. J Control Release. 2007;120(1–2):18–26. Scholar
  30. 30.
    Chen ZN, Mi L, Xu J, Song F, Zhang Q, Zhang Z, et al. Targeting radioimmunotherapy of hepatocellular carcinoma with iodine (131I) metuximab injection: clinical phase I/II trials. Int J Radiat Oncol Biol Phys. 2006;65(2):435–44. Scholar
  31. 31.
    Sun H, Meng F, Cheng R, Deng C, Zhong Z. Reduction-responsive polymeric micelles and vesicles for triggered intracellular drug release. Antioxid Redox Signal. 2014;21(5):755–67. Scholar
  32. 32.
    Zhu WJ, Yang SD, Qu CX, Zhu QL, Chen WL, Li F, et al. Low-density lipoprotein-coupled micelles with reduction and pH dual sensitivity for intelligent co-delivery of paclitaxel and siRNA to breast tumor. Int J Nanomedicine. 2017;12:3375–93. Scholar
  33. 33.
    Zhu R. Study on the preparation and the liver cancer targeting of CD147 antibody-mediated α-Hederin chitosan nanoparticle [dissertation]. Soochow (China): Soochow University; 2014.Google Scholar
  34. 34.
    Oh BM, Lee SJ, Cho HJ, Park YS, Kim JT, Yoon SR, et al. Cystatin SN inhibits auranofin-induced cell death by autophagic induction and ROS regulation via glutathione reductase activity in colorectal cancer. Cell Death Dis. 2017;8(3):e2682. Scholar
  35. 35.
    Moon IJ, Kim KR, Chu HS, Kim SH, Chung WH, Cho YS, et al. N-acetylcysteine and N-nitroarginine methyl ester attenuate carboplatin-induced ototoxicity in dissociated spiral ganglion neuron cultures. Clin Exp Otorhinolaryngol. 2011;4(1):11–7. Scholar
  36. 36.
    Sun CY, Liu Y, Du JZ, Cao ZT, Xu CF, Wang J. Facile generation of tumor-pH-labile linkage-bridged block copolymers for chemotherapeutic delivery. Angew Chem Int Ed Engl. 2016;55(3):1010–4. Scholar
  37. 37.
    Zha Q, Wang X, Cheng X, Fu S, Yang G, Yao W, et al. Acid-degradable carboxymethyl chitosan nanogels via an ortho ester linkage mediated improved penetration and growth inhibition of 3-D tumor spheroids in vitro. Mater Sci Eng, Proc Conf. 2017;78:246–57. Scholar
  38. 38.
    Liu M, Min L, Zhu C, Rao Z, Liu L, Xu W, et al. Preparation, characterization and antioxidant activity of silk peptides grafted carboxymethyl chitosan. Int J Biol Macromol. 2017;104(Pt A):732–8. Scholar
  39. 39.
    Yhee JY, Lee S, Kim K. Advances in targeting strategies for nanoparticles in cancer imaging and therapy. Nano. 2014;6(22):13383–90. Scholar
  40. 40.
    Torchilin V. Tumor delivery of macromolecular drugs based on the EPR effect. Adv Drug Deliv Rev. 2011;63(3):131–5. Scholar
  41. 41.
    He Y, Su Z, Xue L, Xu H, Zhang C. Co-delivery of erlotinib and doxorubicin by pH-sensitive charge conversion nanocarrier for synergistic therapy. J Control Release. 2016;229:80–92. Scholar
  42. 42.
    Hinrichs MJ, Dixit R. Antibody drug conjugates: nonclinical safety considerations. AAPS J. 2015;17(5):1055–64. Scholar
  43. 43.
    Yang SD, Zhu WJ, Zhu QL, Chen WL, Ren ZX, Li F, et al. Binary-copolymer system base on low-density lipoprotein-coupled N-succinyl chitosan lipoic acid micelles for co-delivery MDR1 siRNA and paclitaxel, enhances antitumor effects via reducing drug. J Biomed Mater Res, Part B. 2017;105(5):1114–25. Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2018

Authors and Affiliations

  • Chenxi Qu
    • 1
  • Jizhao Li
    • 1
  • Yejuan Zhou
    • 1
  • Shudi Yang
    • 1
  • Weiliang Chen
    • 1
  • Fang Li
    • 1
  • Bengang You
    • 1
  • Yang Liu
    • 1
  • Xuenong Zhang
    • 1
  1. 1.Department of Pharmaceutics, College of Pharmaceutical Sciences, DuShuHu High Education ZoneSoochow UniversitySuzhouPeople’s Republic of China

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