The present work aimed to develop and evaluate a colloidal system composed of poly (DL-lactide-co-glycolide) (PLGA) nanoparticles (NPs) associated with chlorambucil (CHB) and its effects on cancer cells. The nanoparticles showed %EE (>92%), a mean particle size in the range of 240 to 334 nm and zeta potential of -16.7 to -26.0 mV. In vitro release profile showed a biphasic pattern, with an initial burst for all formulations. The scanning electron microscopy of CHB-nanoparticles showed regular spherical shapes, smooth surface without aggregations. Differential scanning calorimetry thermograms, UV-vis absorption, fluorescence emission and Fourier transform infrared spectroscopy were performed showing the entrapment of the antitumoral in drug delivery system. CHB encapsulated in PLGA nanoparticles decrease the survival rates of the breast cancer cells: 68.9% reduction of cell viability on MCF-7 cell line and 59.7% on NIH3T3. Our results indicated that polymeric nanoparticles produced by classical methods are efficient drug delivery systems for CHB.
References
[1]
Zhang, H., Ye, Z.W. and Lou, Y.J. (2004) Metabolism of Chlorambucil by Rat Liver Microsomal Glutathione S-Transferase. Chemico-Biological Interactions, 149, 61-67. http://dx.doi.org/10.1016/j.cbi.2003.07.002
[2]
Schmidt, M. (2014) Chemotherapy in Early Breast Cancer: When, How and Which One? Breast Care (Basel), 9, 154- 160. http://dx.doi.org/10.1159/000363755
[3]
Cummings, S.R., Tice, J.A., Bauer, S., Browner, W.S., Cuzick, J., Ziv, E., Vogel, V., Shepherd, J., Vachon, C., Smith- Bindman, R. and Kerlikowske, K. (2009) Prevention of Breast Cancer in Postmenopausal Women: Approaches to Estimating and Reducing Risk. Journal of the National Cancer Institute, 101, 384-398.
http://dx.doi.org/10.1093/jnci/djp018
[4]
Zhang, W.J., Feng, M.L., Zheng, G.P., Chen, Y., Wang, X.D., Pen, B., Yin, J., Yu, Y.H. and He, Z.M. (2012) Chemoresistance to 5-Fluorouracil Induces Epithelial-Mesenchymal Transition via Up-Regulation of Snail in MCF7 Human Breast Cancer Cells. Biochemical and Biophysical Research Communications, 417, 679-685.
http://dx.doi.org/10.1016/j.bbrc.2011.11.142
[5]
Bergh, J., Jonsson, P.E., Glimelius, B., Nygren, P. and Care, S.B.-G.S.C.O.T.A.I.H. (2001) A Systematic Overview of Chemotherapy Effects in Breast Cancer. Acta Oncologica, 40, 253-281. http://dx.doi.org/10.1080/02841860151116349
[6]
Anderson, W.F., Rosenberg, P.S., Prat, A., Perou, C.M. and Sherman, M.E. (2014) How Many Etiological Subtypes of Breast Cancer: Two, Three, Four, or More? Journal of the National Cancer Institute, 106, 1-11.
[7]
Pettersson, A., Graff, R.E., Ursin, G., Santos Silva, I.D., McCormack, V., Baglietto, L., Vachon, C., Bakker, M.F., Giles, G.G., Chia, K.S., Czene, K., Eriksson, L., Hall, P., Hartman, M., Warren, R.M., Hislop, G., Chiarelli, A.M., Hopper, J.L., Krishnan, K., Li, J., Li, Q., Pagano, I., Rosner, B.A., Wong, C.S., Scott, C., Stone, J., Maskarinec, G., Boyd, N.F., van Gils, C.H. and Tamimi, R.M. (2014) Mammographic Density Phenotypes and Risk of Breast Cancer: A Meta-Analysis. Journal of the National Cancer Institute, 106, 1-11.
[8]
Bielawski, K., Bielawska, A., Muszynska, A., Poplawska, B. and Czarnomysy, R. (2011) Cytotoxic Activity of G3 PAMAM-NH2 Dendrimer-Chlorambucil Conjugate in Human Breast Cancer Cells. Environmental Toxicology and Pharmacology, 32, 364-372. http://dx.doi.org/10.1016/j.etap.2011.08.002
[9]
Benitah, N., de Lorimier, L.-P., Gaspar, M. and Kitchell, B.E. (2003) Chlorambucil-Induced Myoclonus in a Cat with Lymphoma. Journal of the American Animal Hospital Association, 39, 283-287. http://dx.doi.org/10.5326/0390283
[10]
Hehn, S.T., Dorr, R.T. and Miller, T.P. (2003) Mood Alterations in Patients Treated with Chlorambucil. Clinical lymphoma, 4, 179-182. http://dx.doi.org/10.3816/CLM.2003.n.028
[11]
Reux, B., Weber, V., Galmier, M.J., Borel, M., Madesclaire, M., Madelmont, J.C., Debiton, E. and Coudert, P. (2008) Synthesis and Cytotoxic Properties of New Fluorodeoxyglucose-Coupled Chlorambucil Derivatives. Bioorganic & Medicinal Chemistry, 16, 5004-5020. http://dx.doi.org/10.1016/j.bmc.2008.03.038
[12]
Ganta, S., Paxton, J.W., Baguley, B.C. and Garg, S. (2008) Pharmacokinetics and Pharmacodynamics of Chlorambucil Delivered in Parenteral Emulsion. International Journal of Pharmaceutics, 360, 115-121.
http://dx.doi.org/10.1016/j.ijpharm.2008.04.027
[13]
Yordanov, G.G., Bedzhova, Z.A. and Dushkin, C.D. (2010) Preparation and Physicochemical Characterization of Novel Chlorambucil-Loaded Nanoparticles of Poly(Butylcyanoacrylate). Colloid and Polymer Science, 288, 893-899.
http://dx.doi.org/10.1007/s00396-010-2219-5
[14]
Descoteaux, C., Leblanc, V., Brasseur, K., Gupta, A., Asselin, E. and Berube, G. (2010) Synthesis of D-and L-Tyrosine- Chlorambucil Analogs Active against Breast Cancer Cell Lines. Bioorganic & Medicinal Chemistry Letters, 20, 7388- 7392. http://dx.doi.org/10.1016/j.bmcl.2010.10.039
[15]
Omoomi, F.D. (2013) Molecular Chlorambucil-Methionine Conjugate: Novel Anti-Cancer Agent against Breast MCF-7 Cell Model. Journal of Cancer Science & Therapy, 5, 75-84.
[16]
Millard, M., Gallagher, J.D., Olenyuk, B.Z. and Neamati, N. (2013) A Selective Mitochondrial-Targeted Chlorambucil with Remarkable Cytotoxicity in Breast and Pancreatic Cancers. Journal of Medicinal Chemistry, 56, 9170-9179.
http://dx.doi.org/10.1021/jm4012438
[17]
Gomes, A.J., Barbougli, P.A., Espreafico, E.M. and Tfouni, E. (2008) Trans-[Ru(NO)(NH3)4(py)](BF4)3·H2O Encapsulated in PLGA Microparticles for Delivery of Nitric Oxide to B16-F10 Cells: Cytotoxicity and Phototoxicity. Journal of Inorganic Biochemistry, 102, 757-766. http://dx.doi.org/10.1016/j.jinorgbio.2007.11.012
[18]
Gomes, A.J., Espreafico, E.M. and Tfouni, E. (2013) Trans-[Ru(NO)Cl(Cyclam)](PF6)2 and [Ru(NO)(Hedta)] Incorporated in PLGA Nanoparticles for the Delivery of Nitric Oxide to B16-F10 Cells: Cytotoxicity and Phototoxicity. Molecular Pharmaceutics, 10, 3544-3554.
[19]
Birnbaum, D., Kosmala, J. and Brannon-Peppas, L. (2000) Optimization of Preparation Techniques for Poly(Lactic Acid-Co-Glycolic Acid) Nanoparticles. Journal of Nanoparticle Research, 2, 173-181.
http://dx.doi.org/10.1023/A:1010038908767
[20]
Kranz, H. and Bodmeier, R. (2007) A Novel in Situ Forming Drug Delivery System for Controlled Parenteral Drug Delivery. International Journal of Pharmaceutics, 332, 107-114. http://dx.doi.org/10.1016/j.ijpharm.2006.09.033
[21]
Vandervoort, J. and Ludwig, A. (2002) Biocompatible Stabilizers in the Preparation of PLGA Nanoparticles: A Factorial Design Study. International Journal of Pharmaceutics, 238, 77-92.
http://dx.doi.org/10.1016/S0378-5173(02)00058-3
[22]
Wischke, C. and Schwendeman, S.P. (2008) Principles of Encapsulating Hydrophobic Drugs in PLA/PLGA Microparticles. International Journal of Pharmaceutics, 364, 298-327. http://dx.doi.org/10.1016/j.ijpharm.2008.04.042
[23]
Locatelli, E. and Comes Franchini, M. (2012) Biodegradable PLGA-b-PEG Polymeric Nanoparticles: Synthesis, Properties, and Nanomedical Applications as Drug Delivery System. Journal of Nanoparticle Research, 14, 1-17.
http://dx.doi.org/10.1007/s11051-012-1316-4
[24]
Manchanda, R., Fernandez-Fernandez, A., Nagesetti, A. and McGoron, A.J. (2010) Preparation and Characterization of a Polymeric (PLGA) Nanoparticulate Drug Delivery System with Simultaneous Incorporation of Chemotherapeutic and Thermo-Optical Agents. Colloids and Surfaces B: Biointerfaces, 75, 260-267.
http://dx.doi.org/10.1016/j.colsurfb.2009.08.043
[25]
Danhier, F., Ansorena, E., Silva, J.M., Coco, R., Le Breton, A. and Preat, V. (2012) PLGA-Based Nanoparticles: An Overview of Biomedical Applications. Journal of Controlled Release, 161, 505-522.
http://dx.doi.org/10.1016/j.jconrel.2012.01.043
[26]
Makadia, H.K. and Siegel, S.J. (2011) Poly Lactic-co-Glycolic Acid (PLGA) as Biodegradable Controlled Drug Delivery Carrier. Polymers (Basel), 3, 1377-1397. http://dx.doi.org/10.3390/polym3031377
[27]
Mao, S., Xu, J., Cai, C., Germershaus, O., Schaper, A. and Kissel, T. (2007) Effect of WOW Process Parameters on Morphology and Burst Release of FITC-Dextran Loaded PLGA Microspheres. International Journal of Pharmaceutics, 334, 137-148. http://dx.doi.org/10.1016/j.ijpharm.2006.10.036
[28]
Gomes, A.J., Assuncao, R.M.N., Rodrigues, G., Espreafico, E.M. and Machado, A.E.D. (2007) Preparation and Characterization of Poly(D,L-lactic-co-glycolic Acid) Nanoparticles Containing 3-(Benzoxazol-2-yl)-7-(N,N-diethyl ami- no)chromen-2-one. Journal of Applied Polymer Science, 105, 964-972. http://dx.doi.org/10.1002/app.26204
[29]
Gomes, A.J., Faustino, A.S., Lunardi, C.N., Lunardi, L.O. and Machado, A.E.H. (2007) Evaluation of Nanoparticles Loaded with Benzopsoralen in Rat Peritoneal Exudate Cells. International Journal of Pharmaceutics, 332, 153-160.
http://dx.doi.org/10.1016/j.ijpharm.2006.09.035
[30]
Gomes, A.J., Lunardi, L.O., Marchetti, J.M., Lunardi, C.N. and Tedesco, A.C. (2005) Photobiological and Ultrastructural Studies of Nanoparticles of Poly(lactic-co-glycolic Acid)-Containing Bacteriochlorophyll-a as a Photosensitizer Useful for PDT Treatment. Drug Delivery, 12, 159-164. http://dx.doi.org/10.1080/10717540590931846
[31]
Gomes, A.J., Lunardi, C.N., Lunardi, L.O., Pitol, D.L. and Machado, A.E.H. (2008) Identification of Psoralen Loaded PLGA Microspheres in Rat Skin by Light Microscopy. Micron, 39, 40-44.
http://dx.doi.org/10.1016/j.micron.2007.06.015
[32]
Gomes, A.J., Faustino, A.S., Machado, A.E.H., Zaniquelli, M.E.D., Rigoletto, T.D., Lunardi, C.N. and Lunardi, L.O. (2006) Characterization of PLGA Microparticles as a Drug Carrier for 3-Ethoxycarbonyl-2H-benzofuro[3,2-f]-1-ben- zopyran-2-one. Ultrastructural Study of Cellular Uptake and Intracellular Distribution. Drug Delivery, 13, 447-454.
http://dx.doi.org/10.1080/10717540600640369
[33]
Tfouni, E., Truzzi, D.R., Tavares, A., Gomes, A.J., Figueiredo, L.E. and Franco, D.W. (2012) Biological Activity of Ruthenium Nitrosyl Complexes. Nitric Oxide-Biology and Chemistry, 26, 38-53.
http://dx.doi.org/10.1016/j.niox.2011.11.005
[34]
Tfouni, E., Doro, F.G., Gomes, A.J., da Silva, R.S., Metzker, G., Benini, P.G.Z. and Franco, D.W. (2010) Immobilized Ruthenium Complexes and Aspects of Their Reactivity. Coordination Chemistry Reviews, 254, 355-371.
http://dx.doi.org/10.1016/j.ccr.2009.10.011
[35]
Gomes, A.J., Lunardi, L.O., Caetano, F.H., Machado, A.E.H., Oliveira-Campos, A.M.F., Bendhack, L.M. and Lunardi, C.N. (2011) Biodegradable Nanoparticles Containing Benzopsoralens: An Attractive Strategy for Modifying Vascular Function in Pathological Skin Disorders. Journal of Applied Polymer Science, 121, 1348-1354.
http://dx.doi.org/10.1002/app.33427
[36]
Mo, L., Hou, L., Guo, D., Xiao, X., Mao, P. and Yang, X. (2012) Preparation and Characterization of Teniposide PLGA Nanoparticles and Their Uptake in Human Glioblastoma U87MG Cells. International Journal of Pharmaceutics, 436, 815-824. http://dx.doi.org/10.1016/j.ijpharm.2012.07.050
[37]
Wang, Y.C., Xu, G.L., Jia, W.D., Han, S.J., Ren, W.H., Wang, W., Liu, W.B., Zhang, C.H. and Chen, H. (2012) Estrogen Suppresses Metastasis in Rat Hepatocellular Carcinoma through Decreasing Interleukin-6 and Hepatocyte Growth Factor Expression. Inflammation, 35, 143-149. http://dx.doi.org/10.1007/s10753-011-9299-3
Song, H., Nie, S., Yang, X., Li, N., Xu, H., Zheng, L. and Pan, W. (2010) Characterization and in Vivo Evaluation of Novel Lipid-Chlorambucil Nanospheres Prepared Using a Mixture of Emulsifiers for Parenteral Administration. International Journal of Nanomedicine, 5, 933-942. http://dx.doi.org/10.2147/IJN.S14596
[40]
de Jesus Gomes, A., Lunardi, C.N., Caetano, F.H., Lunardi, L.O. and da Hora Machado, A.E. (2006) Phagocytosis of PLGA Microparticles in Rat Peritoneal Exudate Cells: A Time-Dependent Study. Microscopy and Microanalysis, 12, 399-405. http://dx.doi.org/10.1017/S1431927606060284
[41]
Gupta, H., Aqil, M., Khar, R.K., Ali, A., Bhatnagar, A. and Mittal, G. (2010) Sparfloxacin-Loaded PLGA Nanoparticles for Sustained Ocular Drug Delivery. Nanomedicine: Nanotechnology Biology and Medicine, 6, 324-333.
http://dx.doi.org/10.1016/j.nano.2009.10.004
[42]
Gunasekaran, S., Kumaresan, S., Balaji, R.A., Anand, G. and Seshadri, S. (2008) Vibrational Spectra and Normal Coordinate Analysis on Structure of Chlorambucil and Thioguanine. Pramana: Journal of Physics, 71, 1291-1300.
http://dx.doi.org/10.1007/s12043-008-0183-0
[43]
Yang, J., Lee, C.H., Park, J., Seo, S., Lim, E.K., Song, Y.J., Suh, J.S., Yoon, H.G., Huh, Y.M. and Haam, S. (2007) Antibody Conjugated Magnetic PLGA Nanoparticles for Diagnosis and Treatment of Breast Cancer. Journal of Materials Chemistry, 17, 2695-2699. http://dx.doi.org/10.1039/b702538f
[44]
Garcia, X., Escribano, E., Domenech, J., Queralt, J. and Freixes, J. (2011) In Vitro Characterization and in Vivo Analgesic and Anti-Allodynic Activity of PLGA-Bupivacaine Nanoparticles. Journal of Nanoparticle Research, 13, 2213- 2223. http://dx.doi.org/10.1007/s11051-010-9979-1
[45]
Mi, F.L., Lin, Y.M., Wu, Y.B., Shyu, S.S. and Tsai, Y.H. (2002) Chitin/PLGA Blend Microspheres as a Biodegradable Drug-Delivery System: Phase-Separation, Degradation and Release Behavior. Biomaterials, 23, 3257-3267.
http://dx.doi.org/10.1016/S0142-9612(02)00084-4
[46]
Namazi, H. and Jafarirad, S. (2011) In Vitro Photo-Controlled Drug Release System Based on Amphiphilic Linear-Dendritic Diblock Copolymers; Self-Assembly Behavior and Application as Nanocarrier. Journal of Pharmaceutical Sciences, 14, 162-180.
[47]
Ray, R.S., Rana, B., Swami, B., Venu, V. and Chatterjee, M. (2006) Vanadium Mediated Apoptosis and Cell Cycle Arrest in MCF7 Cell Line. Chemico-Biological Interactions, 163, 239-247. http://dx.doi.org/10.1016/j.cbi.2006.08.006
[48]
Fonseca, C., Simoes, S. and Gaspar, R. (2002) Paclitaxel-Loaded PLGA Nanoparticles: Preparation, Physicochemical Characterization and in Vitro Anti-Tumoral Activity. Journal of Controlled Release, 83, 273-286.
http://dx.doi.org/10.1016/S0168-3659(02)00212-2
[49]
Averineni, R.K.S., Shavi, G.V., Gurram, A.K., Deshpande, P.B., Arumugam, K., Maliyakkal, N., Meka, S.R. and Nayahirama, U. (2012) PLGA 50:50 Nanoparticles of Paclitaxel: Development, in Vitro Anti-Tumor Activity in BT-549 Cells and in Vivo Evaluation. Bulletin of Materials Science, 35, 319-326.
http://dx.doi.org/10.1007/s12034-012-0313-7
[50]
Mukerjee, A. and Vishwanatha, J.K. (2009) Formulation, Characterization and Evaluation of Curcumin-Loaded PLGA Nanospheres for Cancer Therapy. Anticancer Research, 29, 3867-3875.
[51]
Pirooznia, N., Hasannia, S., Lotfi, A.S. and Ghanei, M. (2012) Encapsulation of Alpha-1 Antitrypsin in PLGA Nanoparticles: In Vitro Characterization as an Effective Aerosol Formulation in Pulmonary Diseases. Journal of Nanobiotechnology, 10, 1-15.
[52]
Ribeiro-Costa, R.M., Alves, A.J., Santos, N.P., Nascimento, S.C., Goncalves, E.C., Silva, N.H., Honda, N.K. and Santos-Magalhaes, N.S. (2004) In Vitro and in Vivo Properties of Usnic Acid Encapsulated into PLGA-Microspheres. Journal of Microencapsulation, 21, 371-384. http://dx.doi.org/10.1080/02652040410001673919
