AAPS PharmSciTech

, Volume 19, Issue 5, pp 2395–2406 | Cite as

Co-delivery of Metformin and Paclitaxel Via Folate-Modified pH-Sensitive Micelles for Enhanced Anti-tumor Efficacy

  • Yu Xiao
  • Shuang Wang
  • Qingyu Zong
  • Zongning YinEmail author
Research Article


Single chemotherapeutic agent like paclitaxel (PTX) has shown some limitations in anti-tumor treatment, such as undesirable side effects, multidrug resistance, and high toxicity. In order to reduce the toxicity of PTX and increase the anti-tumor effect, folate-modified amphiphilic and biodegradable biomaterial was developed to co-deliver PTX and metformin (MET) for exerting the synergistic effect. PTX was physically entrapped in the hydrophobic inner core of the amphiphilic block copolymer by a solvent evaporation method, whereas MET was chemically conjugated to the hydrophilic terminals of copolymer via a pH-sensitive cis-aconityl linkage (Cis). The in vitro release behaviors of the drugs were analyzed by high-performance liquid chromatography (HPLC), and the synergistic effect of the drugs was evaluated by a Q value method. Results showed that drug-loaded micelles with an average size about 100 nm were successfully constructed. In acidic environments, the chemically conjugated MET was rapidly released after the breakage of sensitive bond between drug and copolymer. In vitro anti-tumor studies demonstrated that MET and PTX had a synergistic effect and co-delivery micelles induced higher cytotoxicity and apoptosis against 4T1 breast cancer cells than free drugs. Furthermore, folate-targeted co-delivery micelles increased the cellular uptake of drugs and were found to be effective for the treatment of solid tumor in vivo. These findings indicated that co-delivery of MET and PTX through the polymeric micelles is a promising strategy for cancer therapy.


metformin paclitaxel polymeric micelles folic acid pH-sensitive 



We are thankful to the Key Laboratory of Drug Targeting and Drug Delivery Systems for providing the 4T1 cell and the near-infrared fluorescence dye, DiD.

Funding Information

This research was financially supported by the National Natural Science Foundation of China (No. 81673363).


  1. 1.
    Birsoy K, Possemato R, Franziska K, Lorbeer FK, et al. Metabolic determinants of cancer cell sensitivity to glucose limitation and biguanides. Nature. 2014;508(7494):108–12. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Bodmer M, Becker C, Meier C, Jick SS, Meier CR. Use of metformin and the risk of ovarian cancer: a case-control analysis. Gynecol Oncol. 2011;123(2).
  3. 3.
    Evans JM, Donnelly LA, et al. Metformin and reduced risk of cancer in diabetic patients. BMJ. 2005;330(7503):1304–5. Scholar
  4. 4.
    Kalender A, Selvaraj A, Kim SY, Gulati P, Brûlé S, Viollet B, et al. Metformin, independent of AMPK, inhibits mTORC1 in a rag GTPase-dependent manner. Cell Metab. 2010;11(5):390–401. Scholar
  5. 5.
    Rocha GZ, Dias MM, Ropelle ER, Osorio-Costa F, Rossato FA, Vercesi AE, et al. Metformin amplifieschemotherapy-induced AMPK activation and antitumoral growth. Clin Cancer Res. 2011;17(12):3993–4005. Scholar
  6. 6.
    Hanna RK, Zhou C, Malloy KM, Sun L, Zhong Y, Gehrig PA, et al. Metformin potentiates the effects of paclitaxel in endometrial cancer cells through inhibition of cell proliferation and modulation of the mTOR pathway. Gynecol Oncol. 2012;125(2):458–69. Scholar
  7. 7.
    Jin T, Li TJ, Wu T. Antitumor machanism and toxicology of paclitaxel. J Northeast Agric Univ. 2005;36(6):816–9.Google Scholar
  8. 8.
    Yared JA, Tkaczuk KH. Update on taxane development: new analogs and new formulations. Drug Des Devel Ther. 2012;6:371–84. Scholar
  9. 9.
    Wang H, Zhao Y, Wu Y, Hu YL, Nan K, Nie G, et al. Enhanced anti-tumor efficacy by co-delivery of doxorubicin and paclitaxel with amphiphilic methoxy PEG–PLGA copolymer nanoparticles. Biomaterials. 2011;32(32):8281–90. Scholar
  10. 10.
    Ma YK, Fan XH, Li L. pH-sensitive polymeric micelles formed by doxorubicin conjugated prodrugs for co-delivery of doxorubixin and paclitaxel. Carbohydr Polym. 2016;137(10):19–29. Scholar
  11. 11.
    Ma Y, Liu D, Wang D, Wang Y, Fu Q, Fallon JK, et al. Combinational delivery of hydrophobic and hydrophilic anticancer drugs in single nanoemulsions to treat MDR in cancer. Mol Pharm. 2014;11(8):2623–30. Scholar
  12. 12.
    Zhao X, Chen Q, Li Y, et al. Doxorubicin and curcumin co-delivery by lipid nanoparticles for enhanced treatment of diethylnitrosamine-induced hepatocellular carcinomainmice. Eur J Pharm Biopharm. 2015;93:27–36. Scholar
  13. 13.
    Iliopoulos D, Hirsch HA, Struhl K. Metformin decreases the dose of chemotherapy for prolonging tumor remission in mouse xenografts involving multiple cancer cell types. Cancer Res. 2011;71(9):3196–201. Scholar
  14. 14.
    Kim HJ, Kwon H, Lee JW, Kim HJ, Lee SB, Park HS, et al. Metformin increases survival in hormone receptor-positive, HER2-positive breast cancer patients with diabetes. Breast Cancer Res. 2015;17(1):64–78. Scholar
  15. 15.
    Yu G, Fang W, Xia T, Chen Y, Gao Y, Jiao X, et al. Metformin potentiates rapamycin and cisplatin in gastric cancer in mice. Oncotarget. 2015;6(14):12748–62. Scholar
  16. 16.
    Li W, Wang QL, Liu X, Dong SH, Li HX, Li CY, et al. Combined use of vitamin D3 and metformin exhibits synergistic chemopreventive effects on colorectal neoplasia in rats and mice. Cancer Prev Res. 2015;8(2):139–48. Scholar
  17. 17.
    Sharma SV, Bell DW, Settleman J, Haber DA. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer. 2007;7(3):169–81. Scholar
  18. 18.
    Graham GG, Punt J, Arora M, et al. Clinical pharmacokinetics of metformin. Clin Pharmacokinet. 2010;50(2):81–98. Scholar
  19. 19.
    Cetin M, Atila A, Sahin S, Vural I. Preparation and characterization of metformin hydrochloride loaded-EudragitRSPO and EudragitRSPO/PLGA nanoparticles. Pharm Dev Technol. 2013;18(3):570–6. Scholar
  20. 20.
    Marathe PH, Wen Y, Norton J, Greene DS, Barbhaiya RH, Wilding IR. Effect of altered gastric emptying and gastrointestinal motility on metformin absorption. Br J Clin Pharmacol. 2000;50(4):325–32. Scholar
  21. 21.
    Tian Y, Mao SR. Amphiphilic polymeric micelles as the nanocarrier for peroral delivery of poorly soluble anticancer drugs. Expert Opin Drug Deliv. 2012;9(6):687–700. Scholar
  22. 22.
    Zhang JM, Li YB, Gao W, et al. Andrographolide-loaded PLGA–PEG–PLGA micelles to improve its bioavailability and anticancer efficacy. Expert Opin Drug Deliv. 2015;12(4):689. Scholar
  23. 23.
    Greco F, Vicent MJ. Combination therapy: opportunities and challenges for polymer–drug conjugates as anticancer nanomedicines. Adv Drug Deliv Rev. 2009;61(13):1203–13. Scholar
  24. 24.
    Fang XB, Zhang JM. pH-sensitive micelles based on acid-labile pluronic F68–curcumin conjugates for improved tumor intracellular drug delivery. Int J Pharm. 2016;50(2):28–37. Scholar
  25. 25.
    Kumar A, Balakrishna T, Rajiv J, et al. Formulation and evaluation of mucoadhesive microcapsules of metformin HCl with gum karaya. Int J Pharm Pharm Sci. 2011;3:150–5.Google Scholar
  26. 26.
    Choudhury PK, Kar M. Controlled release metformin hydrochloride microspheres of ethyl cellulose prepared by different methods and study on the polymer affected parameters. J Microencapsul. 2009;26(1):46–53. Scholar
  27. 27.
    Snima KS, Jayakumar R, Lakshmanan VK. In vitro and in vivo biological evaluation of o-carboxymethyl chitosan encapsulated metformin nanoparticles for pancreatic cancer therapy. Pharm Res. 2014;31(12):3361–70. Scholar
  28. 28.
    Snima KS, Jayakumar R, Unnikrishnan AG, Nair SV, Lakshmanan VK. Ocarboxymethyl chitosan nanoparticles for metformin delivery to pancreatic cancer cells. Carbohydr Polym. 2012;89(3):1003–7. Scholar
  29. 29.
    Safavy A, Raisch KP, Mantena S, Sanford LL, Sham SW, Krishna NR, et al. Design and development of water-soluble curcumin conjugates as potential anticancer agents. J Med Chem. 2007;50(24):6284–8. Scholar
  30. 30.
    Dey S, Sreenivasan K. Conjugation of curcumin onto alginate enhances aqueous solubility and stability of curcumin. Carbohydr Polym. 2014;99(99):499–507. Scholar
  31. 31.
    Zhao X, Chen Q, Li Y, Tang H, Liu W, Yang X. Doxorubicin and curcumin co-delivery by lipid nanoparticles for enhanced treatment of diethylnitrosamine-induced hepatocellular carcinoma in mice. Eur J Pharm Biopharm. 2015;93:27–36. Scholar
  32. 32.
    Chango A, Nour AA, Bousserouel S, Eveillard D, Anton PM, Guéant JL. Time course gene expression in the one-carbon metabolism network using HepG2 cell line grown in folate-deficient medium. J Nutr Biochem. 2009;20(4):312–20. Scholar
  33. 33.
    Prabaharan M, Grailer JJ, Pilla S, Steeber DA, Gong S. Folate-conjugated amphiphilic hyperbranched block copolymers based on Boltorn H40, poly(L-lactide) and poly(ethylene glycol) for tumor-targeted drug delivery. Biomaterials. 2009;30(16):3009–19. Scholar
  34. 34.
    Zheng Y, Song XR, Darby M, Liang Y, He L, Cai Z, et al. Preparation and characterization of folate-poly (ethylene glycol)-grafted-trimethylchitosan for intracellular transport of protein through folate receptor-mediated endocytosis. J Biotechnol. 2010;145(1):47–53. Scholar
  35. 35.
    Shmeeda H, Amitay Y, Gorin J, Tzemach D, Mak L, Ogorka J, et al. Delivery of zoledronic acid encapsulated in folate-targeted liposome results in potent in vitro cytotoxic activity on tumor cells. J Control Release. 2010;146(1):76–83. Scholar
  36. 36.
    Pan Y, Ren XT, Wang S, Li X, Luo X, Yin Z. Annexin V-conjugated mixed micelles as a potential drug delivery system for targeted thrombolysis. Biomacromolecules. 2017;18(3):865–76. Scholar
  37. 37.
    Cai Y, Sun ZQ, Fang X, Fang X, Xiao F, Wang Y, et al. Synthesis, characterization and anti-cancer activity of Pluronic F-68-curcumin conjugate micelles. Drug Deliv. 2016;23(7):2587–95. Scholar
  38. 38.
    Hu F, Zhang Y, You J, et al. pH triggered doxorubicin delivery of PEGylated glycolipid conjugate micelles for tumor targeting therapy. Mol Pharm. 2012;9(9):2469–78. Scholar
  39. 39.
    He YW, Wang HS, Zeng J, Fang X, Chen HY, du J, et al. Sodium butyrate inhibits interferon-gamma induced indoleamine 2,3-dioxygenase expression via STAT1 in nasopharyngeal caecinoma cells. Life Sci. 2013;93(15):509–15. Scholar
  40. 40.
    Yang Z, Sun N, Cheng R, Zhao C, Liu Z, Li X, et al. pH multistage responsive micellar system with charge-switch and PEGlayer detachment for co-delivery of paclitaxel and curcumin to synergistically eliminate breast cancer stem cells. Biomaterials. 2017;147:53–67. Scholar
  41. 41.
    Hou J, Guo C, Shi Y, Liu E, Dong W, Yu B, et al. A novel high drug loading mussel-inspired polydopamine hybrid nanoparticle as a pH-sensitive vehicle for drug delivery. Int J Pharm. 2017;533(1):73–83. Scholar
  42. 42.
    Li Y, Xiao Y, Yin Z. Enhanced anti-inflammatory efficacy through targeting to macrophages: synthesis and in vitro evaluation of folate-glycine-celecoxib. AAPS PharmSciTech. 2016;18(3):1–9. Scholar
  43. 43.
    Shi J, Kantoff PW, Wooster R, Farokhzad OC. Cancer nanomedicine: progress, challenges and opportunities. Nat Rev Cancer. 2017;17(1):20–37. Scholar
  44. 44.
    Ke XY, Ng V, Gao SJ, et al. Co-delivery of thioridazine and doxorubicin using polymeric micelles for targeting both cancer cells and cancer stem cells. Biomaterials. 2014;35(3):1096–108. Scholar
  45. 45.
    Kataoka Y, Takeichi M, Uemura T. Developmental roles and molecular characterization of a drosophila homologue of arabidopsis argonaute1, the founder of a novel gene superfamily. Genes Cells. 2001;6(4):313–25. Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2018

Authors and Affiliations

  • Yu Xiao
    • 1
  • Shuang Wang
    • 1
  • Qingyu Zong
    • 1
  • Zongning Yin
    • 1
    Email author
  1. 1.Key Laboratory of Drug Targeting and Drug Delivery Systems, West China School of PharmacySichuan UniversityChengduChina

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