Doxorubicin-loaded PLGA nanoparticles by nanoprecipitation: preparation, characterization and in vitro evaluation.

AIMS The lack of specificity of chemotherapeutic agents to cancer tissue commonly leads to dose-limiting side effects and poor therapeutic results. Drug delivery systems promise to improve the deficiencies of chemotherapeutic treatment by modifying the biodistribution and pharmacokinetics of the drug in vivo. Here, we report the preparation, characterization and in vitro evaluation of a carrier for the chemotherapeutic drug doxorubicin based on acid-capped poly(lactic-co-glycolic acid) nanoparticles. METHODS Doxorubicin-loaded nanoparticles were prepared by nanoprecipitation with bovine serum albumin as the stabilizer. Nanoparticles were characterized and their interaction with MDA-MB-231 breast cancer cells was examined with confocal microscopy and a toxicological assay. RESULTS Spherical particles with an average diameter of 230 nm, a zeta-potential of -45 mV and a maximum drug loading of 5 wt% were prepared. Doxorubicin was found to be quickly released at endolysosomal pH of 4.0 but was released at a slower rate at pH 7.4. Nanoparticles were found to deliver the drug into cells quickly and in higher quantity than when presented in solution and were found to result in a therapeutic efficacy comparable to the free drug. DISCUSSION Nanoprecipitation was found to be a promising method for the preparation of nanoparticles with relatively high doxorubicin loading. The pH-dependent release behavior is discussed to possibly be a result of accelerated degradation of the polymer and decreasing ionic interaction between the drug and the polymer at acidic pH. Additional studies are needed to determine why increased nuclear localization of the drug when delivered in the form of nanoparticles did not result in increased therapeutic efficacy in vitro. CONCLUSION Nanoparticles formulated by nanoprecipitation of acid-ended poly(lactic-co-glycolic acid) were found to be able to control the release of doxorubicin in a pH-dependent manner and to effectively deliver high payloads of the drug in an active form to MDA-MB-231 breast cancer cells.

[1]  C. Ropert,et al.  Body distribution of fully biodegradable [14C]-poly(lactic acid) nanoparticles coated with albumin after parenteral administration to rats. , 1992, Biomaterials.

[2]  P. Legrand,et al.  Polymeric nanocapsules as drug delivery systems. A review , 1999 .

[3]  P. Couvreur,et al.  Adsorption/desorption of human serum albumin at the surface of poly(lactic acid) nanoparticles prepared by a solvent evaporation process. , 1993, Journal of biomedical materials research.

[4]  Allan G. A. Coombes,et al.  Surface Modification of Poly(lactide-co-glycolide) Nanospheres by Biodegradable Poly(lactide)-Poly(ethylene glycol) Copolymers , 1994, Pharmaceutical Research.

[5]  D. McDonald,et al.  Significance of blood vessel leakiness in cancer. , 2002, Cancer research.

[6]  T. Okano,et al.  Targetable drug carriers: present status and a future perspective , 1996 .

[7]  Chi‐Hwa Wang,et al.  In vitro study of anticancer drug doxorubicin in PLGA-based microparticles. , 2005, Biomaterials.

[8]  M. Morishita,et al.  Encapsulation of hydrophilic and lipophilic drugs in PLGA nanoparticles by the nanoprecipitation method. , 1999, Drug development and industrial pharmacy.

[9]  Hans P Merkle,et al.  Microencapsulation by solvent extraction/evaporation: reviewing the state of the art of microsphere preparation process technology. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[10]  T. Park,et al.  Biodegradable Nanoparticles Containing Doxorubicin-PLGA Conjugate for Sustained Release , 1999, Pharmaceutical Research.

[11]  A. Gabizon.  Pegylated Liposomal Doxorubicin: Metamorphosis of an Old Drug into a New Form of Chemotherapy , 2001, Cancer investigation.

[12]  V. Torchilin.  STRATEGIES AND MEANS FOR DRUG TARGETING: AN OVERVIEW , 2002 .

[13]  F. Torti,et al.  Doxorubicin: Alteration of Dose Scheduling as a Means of Reducing Cardiotoxicity , 1985, Drug Intelligence & Clinical Pharmacy.

[14]  Lisbeth Illum,et al.  Polymers in Controlled Drug Delivery , 1988 .

[15]  P. Legrand,et al.  Poly(D,L-lactide) nanocapsules prepared by a solvent displacement process: influence of the composition on physicochemical and structural properties. , 2000, Journal of pharmaceutical sciences.

[16]  M. A. Arangoa,et al.  Modulation of the cellular immune response after oral or subcutaneous immunization with microparticles containing Brucella ovis antigens. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[17]  K. Janes,et al.  Chitosan nanoparticles as delivery systems for doxorubicin. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[18]  P. Couvreur,et al.  Biodegradable microparticles for the mucosal delivery of antibacterial and dietary antigens. , 2002, International journal of pharmaceutics.

[19]  J. Folkman,et al.  ISOLATION OF A TUMOR FACTOR RESPONSIBLE FOR ANGIOGENESIS , 1971, The Journal of experimental medicine.

[20]  Hyun-chul Lee,et al.  Adriamycin release from self-assembling nanospheres of poly(DL-lactide-co-glycolide)-grafted pullulan. , 2006, International journal of pharmaceutics.

[21]  Michel Veillard,et al.  Non-stealth (poly(lactic acid/albumin)) and stealth (poly(lactic acid-polyethylene glycol)) nanoparticles as injectable drug carriers , 1995 .

[22]  Jayanth Panyam,et al.  Fluorescence and electron microscopy probes for cellular and tissue uptake of poly(D,L-lactide-co-glycolide) nanoparticles. , 2003, International journal of pharmaceutics.

[23]  K. Margolin,et al.  Targeting Vascular Endothelium with Antibodies , 2002 .

[24]  Y. Ikada,et al.  Biodegradation and antitumour effect of adriamycin-containing poly(L-lactic acid) microspheres. , 1991, Biomaterials.

[25]  Tae Gwan Park,et al.  Folate receptor targeted biodegradable polymeric doxorubicin micelles. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[26]  J. Folkman,et al.  Fundamental concepts of the angiogenic process. , 2003, Current molecular medicine.

[27]  M. Shive,et al.  Biodegradation and biocompatibility of PLA and PLGA microspheres , 1997 .

[28]  M. Alonso,et al.  Protein encapsulation and release from poly(lactide-co-glycolide) microspheres: effect of the protein and polymer properties and of the co-encapsulation of surfactants. , 1998, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[29]  S. Schulman,et al.  Electronic absorption spectra and protolytic equilibria of doxorubicin: direct spectrophotometric determination of microconstants. , 1977, Journal of pharmaceutical sciences.

[30]  C. Astete,et al.  Synthesis and characterization of PLGA nanoparticles , 2006, Journal of biomaterials science. Polymer edition.

[31]  K. Akiyoshi,et al.  Hydrogel nanoparticle formed by self-assembly of hydrophobized polysaccharide. Stabilization of adriamycin by complexation , 1996 .

[32]  L. Brannon-Peppas,et al.  Molecular weight distribution changes during degradation and release of PLGA nanoparticles containing epirubicin HCl , 2003, Journal of biomaterials science. Polymer edition.

[33]  Jian Ji,et al.  Novel biomimetic polymersomes as polymer therapeutics for drug delivery. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[34]  Si-Shen Feng,et al.  Folate-decorated poly(lactide-co-glycolide)-vitamin E TPGS nanoparticles for targeted drug delivery. , 2007, Biomaterials.

[35]  L. Brannon-Peppas,et al.  Optimization of Preparation Techniques for Poly(Lactic Acid-Co-Glycolic Acid) Nanoparticles , 2000 .

[36]  A. P. Chapman,et al.  PEGylated antibodies and antibody fragments for improved therapy: a review. , 2002, Advanced drug delivery reviews.

[37]  Jayanth Panyam,et al.  Biodegradable nanoparticles for drug and gene delivery to cells and tissue. , 2003, Advanced drug delivery reviews.

[38]  M. Chevallier,et al.  Increase of doxorubicin sensitivity by doxorubicin-loading into nanoparticles for hepatocellular carcinoma cells in vitro and in vivo. , 2005, Journal of hepatology.

[39]  A. Lamprecht,et al.  In Vitro Evaluation of Betamethasone-Loaded Nanoparticles , 2005, Drug development and industrial pharmacy.

[40]  V. Labhasetwar,et al.  Characterization of nanoparticle uptake by endothelial cells. , 2002, International journal of pharmaceutics.

[41]  T. Park,et al.  Biodegradable polymeric micelles composed of doxorubicin conjugated PLGA-PEG block copolymer. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[42]  J. Błasiak,et al.  A comparison of the in vitro genotoxicity of anticancer drugs idarubicin and mitoxantrone. , 2002, Acta biochimica Polonica.

[43]  Patrick Couvreur,et al.  Controlled drug delivery with nanoparticles : current possibilities and future trends , 1995 .

[44]  G. Tosi,et al.  Nanoparticle formulation may affect the stabilization of an antiischemic prodrug. , 2006, International journal of pharmaceutics.

[45]  Lisa Brannon-Peppas,et al.  Recent advances on the use of biodegradable microparticles and nanoparticles in controlled drug delivery , 1995 .

[46]  You Han Bae,et al.  Doxorubicin loaded pH-sensitive polymeric micelles for reversal of resistant MCF-7 tumor. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[47]  Hua Ai,et al.  Multifunctional polymeric micelles as cancer-targeted, MRI-ultrasensitive drug delivery systems. , 2006, Nano letters.

[48]  Jean-Pierre Benoit,et al.  Physico-chemical stability of colloidal lipid particles. , 2003, Biomaterials.

[49]  R. A. Jain,et al.  The manufacturing techniques of various drug loaded biodegradable poly(lactide-co-glycolide) (PLGA) devices. , 2000, Biomaterials.