Targeting angiogenesis is a strategy to better control tumor growth and metastasis. αvβ3 is an integrin, involved in the regulation of angiogenesis and overexpressed in angiogenic endothelial cells and various cancers including breast, prostate, pancreatic, and brain cancers. cRGDfK peptide has high specificity towards αvβ3 integrin receptors. Docetaxel (DTX) is a broad spectrum anticancer drug, widely used to treat breast, ovarian, prostate, non-small-cell lung, gastric, and neck cancers. Its clinical application is limited owing to its poor aqueous solubility, low oral bioavailability, and nonspecific cytotoxicity. The nanocarriers help to overcome these limitations and further can be surface-modified to conjugate ligand to achieve selective delivery to tumor. d-α-Tocopheryl polyethylene glycol succinate (TPGS) is a water soluble derivative of natural d-α-tocopherol (Vit E). TPGS-based engineered nanocarrier systems have been shown to transport and deliver anticancer drugs more efficiently than the pristine drugs. Herein, we attempt to improve the therapeutic potential of DTX and to target the integrin receptor overexpressing angiogenic tumors, by encapsulating the DTX in nanomicelles and conjugating to cRGDfK peptide for tumor targeting. These nanomicelles are characterized by various analytical techniques and their potential of selective targeting is also evaluated. In the present chapter, we provide the general procedure used in this study: (1) synthesis and characterization of succinoyl-TPGS, (2) preparation and characterization of docetaxel loaded TPSA nanomicelles (DNM), (3) bioconjugation, quantification, and characterization of cRGDfK peptide to DNM (PDNM), (4) in vitro evaluation of cytotoxicity of the nanoparticles, (5) antiangiogenic activity, and (6) stability studies.
Nanomicelles Docetaxel Angiogenesis TPGS Bioconjugation Targeted delivery Cancer therapy
This is a preview of subscription content, log in to check access.
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
H.K. and T.S. Reddy are thankful to the Director, IICT-RMIT Research Centre for providing the Junior Research Fellowships. D.P. and A.K.B. acknowledge the Council of Scientific and Industrial Research (CSIR), New Delhi and the University Grants Commission (UGC), New Delhi, respectively, for awarding Senior Research Fellowships. This work is partially supported by a CSIR grant under project Advanced Drug Delivery Systems (CSC 0302). D.J.A. is an Australian Research Council (ARC) Australian Professorial Fellow.
Declaration: The work described in this book chapter has been published as Kulhari et al., Cyclic-RGDfK peptide conjugated succinoyl-TPGS nanomicelles for targeted delivery of docetaxel to integrin receptor overexpressing angiogenic tumors. Nanomedicine. 2015;11(6):1511–1520. The work is reprinted after permission from Elsevier under license number 3671851307186.
Zhaofei L, Fan W, Xiaoyuan C (2008) Integrin αvβ3-targeted cancer therapy. Drug Dev Res 69:329–339CrossRefGoogle Scholar
Thobe MN, Gurusamy D, Pathrose P, Waltz SE (2010) The Ron receptor tyrosine kinase positively regulates angiogenic chemokine production in prostate cancer cells. Oncogene 29:214–226CrossRefPubMedGoogle Scholar
Cho K, Wang X, Nie S, Chen ZG, Shin DM (2008) Therapeutic nanoparticles for drug delivery in cancer. Clin Cancer Res 14:1310–1316CrossRefPubMedGoogle Scholar
Baker J, Ajani J, Scotte F, Winther D, Martin M, Aapro MS et al (2009) Docetaxel-related side effects and their management. Eur J Oncol Nurs 13:49–59CrossRefPubMedGoogle Scholar
Roy A, Murakami M, Ernsting MJ, Hoang B, Undzys E, Li SD (2014) Carboxymethylcellulose-based and docetaxel-loaded nanoparticles circumvent P-glycoprotein-mediated multidrug resistance. Mol Pharm 11:2592–2599CrossRefPubMedPubMedCentralGoogle Scholar
Kulhari H, Pooja D, Shrivastava S, Naidu VGM, Sistla R (2014) Peptide conjugated polymeric nanoparticles as a carrier for targeted delivery of docetaxel. Colloids Surf B Biointerfaces 117:166–173CrossRefPubMedGoogle Scholar
Pooja D, Kulhari H, Tunki L, Chinde S, Kuncha M, Grover P, Rachamalla SS, Sistla R (2015) Nanomedicines for targeted delivery of etoposide to non-small cell lung cancer using transferrin functionalized nanoparticles. RSC Adv 5:49122–49131CrossRefGoogle Scholar
Sultana S, Khan MR, Kumar M, Kumar S, Ali M (2013) Nanoparticles-mediated drug delivery approaches for cancer targeting: a review. J Drug Target 21:107–125CrossRefPubMedGoogle Scholar
Farrell D, Ptak K, Panaro NJ, Grodzinski P (2011) Nanotechnology-based cancer therapeutics-promise and challenge-lessons learned through the NCI Alliance for nanotechnology in cancer. Pharm Res 28:273–278CrossRefPubMedGoogle Scholar
Kulhari H, Deep Pooja Singh MK, Chauhan AS (2013) Optimization of carboxylate-terminated poly(amidoamine) dendrimer-mediated cisplatin formulation. Drug Dev Ind Pharm 41(2):232–238CrossRefPubMedGoogle Scholar
Guo Y, Luo J, Tan S, Otieno BO, Zhang Z (2013) The applications of Vitamin E TPGS in drug delivery. Eur J Pharm Sci 49:175–186CrossRefGoogle Scholar
Zhang Z, Tan S, Feng SS (2012) Vitamin E TPGS as a molecular biomaterial for drug delivery. Biomaterials 33:4889–4906CrossRefGoogle Scholar
Mi Y, Zhao J, Feng SS (2012) Vitamin E TPGS prodrug micelles for hydrophilic drug delivery with neuroprotective effects. Int J Pharm 438:98–106CrossRefGoogle Scholar
Pooja D, Kulhari H, Singh MK, Mukherjee S, Rachamalla SS, Sistla R (2014) Dendrimer-TPGS mixed micelles for enhanced solubility and cellular toxicity of taxanes. Colloids Surf B Biointerfaces 121:461–468CrossRefGoogle Scholar
Duhem N, Rolland J, Riva R, Guillet P, Schumers JM, Jérome C, Gohy JF, Préat V (2011) Tocol modified glycol chitosan for the oral delivery of poorly soluble drugs. Int J Pharm 423:452–460CrossRefPubMedGoogle Scholar
Schmitt A, Schmitt J, Münch G, Gasic-Milencovic J (2005) Characterization of advanced glycation end products for biochemical studies: side chain modifications and fluorescence characteristics. Anal Biochem 15:201–215CrossRefGoogle Scholar
VanMeerloo J, Kaspers GJL, Cloos J (2011) Cell sensitivity assays: the MTT assay. Cancer cell culture. Springer, New York, pp 237–245CrossRefGoogle Scholar
Gao H, Yang Z, Zhang S, Cao S, Shen S, Pang Z, Jiang X (2013) Ligand modified nanoparticles increases cell uptake, alters endocytosis and elevates glioma distribution and internalization. Sci Rep 3:2534CrossRefPubMedPubMedCentralGoogle Scholar
Oommen S, Anto RJ, Srinivas G, Karunagaran D (2004) Allicin (from garlic) induces caspase-mediated apoptosis in cancer cells. Eur J Pharmacol 485(1):97–103CrossRefPubMedGoogle Scholar
Rieger AM, Nelson KL, Konowalchuk JD, Barreda DR (2011) Modified annexin V/propidium iodide apoptosis assay for accurate assessment of cell death. J Vis Exp 50:2597Google Scholar
Nagababu P, Barui AK, Bathini T (2015) Anti-angiogenic activity of mononuclear copper(II) polypyridyl complexes for the treatment of cancers. J Med Chem 58(13):5226–5241CrossRefPubMedGoogle Scholar
Barui AK, Veeriah V, Mukherjee S et al (2012) Zinc oxide nanoflowers make new blood vessels. Nanoscale 4:7861–7869CrossRefPubMedGoogle Scholar