Synthesis of a Smart Nanovehicle for Targeting Liver

  • Arnab De
  • Sushil Mishra
  • Seema Garg
  • Subho Mozumdar
Part of the Methods in Molecular Biology book series (MIMB, volume 1141)


A protocol for the synthesis of a smart drug delivery system based on gold nanoparticles has been described in this chapter. The synthesized drug delivery system has been shown to release the bioactive material in response to an intracellular stimulus (glutathione concentration gradient) and hence shown to behave in an intelligent manner. Gold nanoparticles have been employed as the core material with the surface functionalities of thiolated PEG. PEG owing to its non-immunogenicity and non-antigenicity would impart considerable stability and longer in vivo circulation time to the gold nanoparticles. The end groups of PEG chains have been derivatized with functional groups like aldehyde (–CHO) and amine (–NH2) which could behave as flexible arms for the attachment of “target specific ligands” and other bioactive substances. Lactose, a liver targeting ligand, has been employed as the target specific moiety. A Coumarin derivative has been synthesized and used as the model fluorescent tag as well as a linker to examine the glutathione-mediated release through fluorescence spectroscopy and for the conjugation of bioactive molecules, respectively. A check for the cytocompatibility of the synthesized nanovehicle on the cultured mammalian cell lines has also been carried out. Finally, in the latter parts of the chapter (mimicking the in vivo conditions), time-dependent in vitro release of the model fluorescent moiety has also been analyzed at different glutathione concentrations.

Key words

Smart drug delivery system Targeting liver Gold nanoparticles Thiolated PEG Lactose Coumarin Glutathione Fluorescence spectroscopy 



The authors thank the Department of Science and Technology (DST), New Delhi for the financial assistance in the form of Junior Research Fellowship and the University Science Instrumentation Centre, University of Delhi, for providing the characterization facilities. The authors also thank Dr. Y. Singh (Scientist “G”, IGIB, Delhi) for carrying out the cytotoxicity work in his lab.


  1. 1.
    Shenoy D, Fu W, Li J, Crasto C, Jones G, DiMarzio C, Sridhar S, Amiji M (2006) Surface functionalization of gold nanoparticles using hetero-bifunctional poly(ethylene glycol) spacer for intracellular tracking and delivery. Int J Nanomedicine 1:51CrossRefGoogle Scholar
  2. 2.
    Gerasimov OV, Boomer JA, Qualls MM, Thompson DH (1999) Cytosolic drug delivery using pH- and light-sensitive liposomes. Adv Drug Deliv Rev 38:317–338CrossRefGoogle Scholar
  3. 3.
    Alvarez-Lorenzo C, Bromberg L, Concheiro A (2009) Light-sensitive intelligent drug delivery systems. Photochem Photobiol 85:848–860CrossRefGoogle Scholar
  4. 4.
    Sawahata K, Hara M, Yasunaga H, Osada Y (1990) Electrically controlled drug delivery system using polyelectrolyte gels. J Control Release 14:253–262CrossRefGoogle Scholar
  5. 5.
    Kwon IC, Bae YH, Okano T, Kim SW (1991) Drug release from electric current sensitive polymers. J Control Release 17:149–156CrossRefGoogle Scholar
  6. 6.
    Yuk SH, Cho SH, Lee HB (1992) Electric current-sensitive drug delivery systems using sodium alginate/polyacrylic acid composites. Pharm Res 9:955–957CrossRefGoogle Scholar
  7. 7.
    Kost J, Leong K, Langer R (1998) Ultrasonically controlled polymeric drug delivery. Paper presented at Makromolekulare Chemie, Macromolecular Symposia 1988Google Scholar
  8. 8.
    Kost J, Leong K, Langer R (1989) Ultrasound-enhanced polymer degradation and release of incorporated substances. Proc Natl Acad Sci U S A 86:7663–7666CrossRefGoogle Scholar
  9. 9.
    Satturwar P, Eddine MN, Ravenelle F, Leroux J-C (2007) pH-responsive polymeric micelles of poly (ethylene glycol)-b-poly (alkyl (meth) acrylate-co-methacrylic acid): influence of the copolymer composition on self-assembling properties and release of candesartan cilexetil. Eur J Pharm Biopharm 65:379–387CrossRefGoogle Scholar
  10. 10.
    Na K, Lee KH, Bae YH (2004) pH-sensitivity and pH-dependent interior structural change of self-assembled hydrogel nanoparticles of pullulan acetate/oligo-sulfonamide conjugate. J Control Release 97:513–525CrossRefGoogle Scholar
  11. 11.
    Chen S-C, Wu Y-C, Mi F-L, Lin Y-H, Yu L-C, Sung H-W (2004) A novel pH-sensitive hydrogel composed of N,O-carboxymethyl chitosan and alginate cross-linked by genipin for protein drug delivery. J Control Release 96:285–300CrossRefGoogle Scholar
  12. 12.
    Hrubý M, Koňák Č, Ulbrich K (2005) Polymeric micellar pH-sensitive drug delivery system for doxorubicin. J Control Release 103:137–148CrossRefGoogle Scholar
  13. 13.
    Ito Y, Casolaro M, Kono K, Imanishi Y (1989) An insulin-releasing system that is responsive to glucose. J Control Release 10:195–203CrossRefGoogle Scholar
  14. 14.
    Shiino D, Murata Y, Kataoka K, Koyama Y, Yokoyama M, Okano T, Sakurai Y (1994) Preparation and characterization of a glucose-responsive insulin-releasing polymer device. Biomaterials 15:121–128CrossRefGoogle Scholar
  15. 15.
    Hisamitsu I, Kataoka K, Okano T, Sakurai Y (1997) Glucose-responsive gel from phenylborate polymer and poly(vinyl alcohol): prompt response at physiological pH through the interaction of borate with amino group in the gel. Pharm Res 14:289–293CrossRefGoogle Scholar
  16. 16.
    Dong-June C, Yoshihiro I, Yukio I (1992) An insulin-releasing membrane system on the basis of oxidation reaction of glucose. J Control Release 18:45–53CrossRefGoogle Scholar
  17. 17.
    Ulijn RV (2006) Enzyme-responsive materials: a new class of smart biomaterials. J Mater Chem 16:2217–2225CrossRefGoogle Scholar
  18. 18.
    Thornton PD, McConnell G, Ulijn RV (2005) Enzyme responsive polymer hydrogel beads. Chem Commun 5913–5915Google Scholar
  19. 19.
    Toledano S, Williams RJ, Jayawarna V, Ulijn RV (2006) Enzyme-triggered self-assembly of peptide hydrogels via reversed hydrolysis. J Am Chem Soc 128:1070–1071CrossRefGoogle Scholar
  20. 20.
    Miyata T, Asami N, Uragami T (1999) Preparation of an antigen-sensitive hydrogel using antigen-antibody bindings. Macromolecules 32:2082–2084CrossRefGoogle Scholar
  21. 21.
    Lu ZR, Kopečková P, Kopeček J (2003) Antigen responsive hydrogels based on polymerizable antibody Fab′ fragment. Macromol Biosci 3:296–300CrossRefGoogle Scholar
  22. 22.
    Zhang R, Bowyer A, Eisenthal R, Hubble J (2007) A smart membrane based on an antigen-responsive hydrogel. Biotechnol Bioeng 97:976–984CrossRefGoogle Scholar
  23. 23.
    Koo AN, Lee HJ, Kim SE, Chang JH, Park C, Kim C, Park JH, Lee SC (2008) Disulfide-cross-linked PEG-poly (amino acid)s copolymer micelles for glutathione-mediated intracellular drug delivery. Chem Commun 6570–6572Google Scholar
  24. 24.
    Tsarevsky NV, Matyjaszewski K (2005) Combining atom transfer radical polymerization and disulfide/thiol redox chemistry: a route to well-defined (bio)degradable polymeric materials. Macromolecules 38:3087–3092CrossRefGoogle Scholar
  25. 25.
    Aerry S, De A, Kumar A, Saxena A, Majumdar D, Mozumdar S (2012) Synthesis and characterization of thermoresponsive copolymers for drug delivery. J Biomed Mater Res A 101A:2015–2026CrossRefGoogle Scholar
  26. 26.
    Bae YH, Okano T, Hsu R, Kim SW (1987) Thermo‐sensitive polymers as on‐off switches for drug release. Makromol Chem Rapid Commun 8:481–485CrossRefGoogle Scholar
  27. 27.
    Chung JE, Yokoyama M, Yamato M, Aoyagi T, Sakurai Y, Okano T (1999) Thermo-responsive drug delivery from polymeric micelles constructed using block copolymers of poly(N-isopropylacrylamide) and poly(butylmethacrylate). J Control Release 62:115–127CrossRefGoogle Scholar
  28. 28.
    Li Y, Pan S, Zhang W, Du Z (2009) Novel thermo-sensitive core-shell nanoparticles for targeted paclitaxel delivery. Nanotechnology 20:065104CrossRefGoogle Scholar
  29. 29.
    Hong R, Han G, Fernández JM, Kim B, Forbes NS, Rotello VM (2006) Glutathione-mediated delivery and release using monolayer protected nanoparticle carriers. J Am Chem Soc 128:1078–1079CrossRefGoogle Scholar
  30. 30.
    Jones DP, Carlson JL, Mody VC Jr, Cai J, Lynn MJ, Sternberg P Jr (2000) Redox state of glutathione in human plasma. Free Radic Biol Med 28:625–635CrossRefGoogle Scholar
  31. 31.
    Anderson ME (1998) Glutathione: an overview of biosynthesis and modulation. Chem Biol Interact 111:1–14CrossRefGoogle Scholar
  32. 32.
    Chompoosor A, Han G, Rotello VM (2008) Charge dependence of ligand release and monolayer stability of gold nanoparticles by biogenic thiols. Bioconjug Chem 19:1342–1345CrossRefGoogle Scholar
  33. 33.
    Han G, Chari NS, Verma A, Hong R, Martin CT, Rotello VM (2005) Controlled recovery of the transcription of nanoparticle-bound DNA by intracellular concentrations of glutathione. Bioconjug Chem 16:1356–1359CrossRefGoogle Scholar
  34. 34.
    Li D, Li G, Guo W, Li P, Wang E, Wang J (2008) Glutathione-mediated release of functional plasmid DNA from positively charged quantum dots. Biomaterials 29:2776–2782CrossRefGoogle Scholar
  35. 35.
    Liu J, Pang Y, Huang W, Huang X, Meng L, Zhu X, Zhou Y, Yan D (2011) Bioreducible micelles self-assembled from amphiphilic hyperbranched multiarm copolymer for glutathione-mediated intracellular drug delivery. Biomacromolecules 12:1567–1577CrossRefGoogle Scholar
  36. 36.
    Sun H, Guo B, Cheng R, Meng F, Liu H, Zhong Z (2009) Biodegradable micelles with sheddable poly(ethylene glycol) shells for triggered intracellular release of doxorubicin. Biomaterials 30:6358–6366CrossRefGoogle Scholar
  37. 37.
    Tang L-Y, Wang Y-C, Li Y, Du J-Z, Wang J (2009) Shell-detachable micelles based on disulfide-linked block copolymer as potential carrier for intracellular drug delivery. Bioconjug Chem 20:1095–1099CrossRefGoogle Scholar
  38. 38.
    Yang P-H, Sun X, Chiu J-F, Sun H, He Q-Y (2005) Transferrin-mediated gold nanoparticle cellular uptake. Bioconjug Chem 16:494–496CrossRefGoogle Scholar
  39. 39.
    Han G, Ghosh P, Rotello VM (2007) Functionalized gold nanoparticles for drug delivery. Nanomedicine 2:113–123CrossRefGoogle Scholar
  40. 40.
    Pingarrón JM, Yáñez-Sedeño P, González-Cortés A (2008) Gold nanoparticle-based electrochemical biosensors. Electrochim Acta 53:5848–5866CrossRefGoogle Scholar
  41. 41.
    El-Sayed IH, Huang X, El-Sayed MA (2005) Surface plasmon resonance scattering and absorption of anti-EGFR antibody conjugated gold nanoparticles in cancer diagnostics: applications in oral cancer. Nano Lett 5:829–834CrossRefGoogle Scholar
  42. 42.
    Shukla S, Priscilla A, Banerjee M, Bhonde RR, Ghatak J, Satyam P, Sastry M (2005) Porous gold nanospheres by controlled transmetalation reaction: a novel material for application in cell imaging. Chem Mater 17:5000–5005CrossRefGoogle Scholar
  43. 43.
    Chen J, Saeki F, Wiley BJ, Cang H, Cobb MJ, Li Z-Y, Au L, Zhang H, Kimmey MB, Li X (2005) Gold nanocages: bioconjugation and their potential use as optical imaging contrast agents. Nano Lett 5:473–477CrossRefGoogle Scholar
  44. 44.
    Cheng MM-C, Cuda G, Bunimovich YL, Gaspari M, Heath JR, Hill HD, Mirkin CA, Nijdam AJ, Terracciano R, Thundat T (2006) Nanotechnologies for biomolecular detection and medical diagnostics. Curr Opin Chem Biol 10:11–19CrossRefGoogle Scholar
  45. 45.
    Rosi NL, Mirkin CA (2005) Nanostructures in biodiagnostics. Chem Rev 105:1547–1562CrossRefGoogle Scholar
  46. 46.
    Pissuwan D, Niidome T, Cortie MB (2011) The forthcoming applications of gold nanoparticles in drug and gene delivery systems. J Control Release 149:65–71CrossRefGoogle Scholar
  47. 47.
    Ghosh P, Han G, De M, Kim CK, Rotello VM (2008) Gold nanoparticles in delivery applications. Adv Drug Deliv Rev 60:1307–1315CrossRefGoogle Scholar
  48. 48.
    Gu Y-J, Cheng J, Lin C-C, Lam YW, Cheng SH, Wong W-T (2009) Nuclear penetration of surface functionalized gold nanoparticles. Toxicol Appl Pharmacol 237:196–204CrossRefGoogle Scholar
  49. 49.
    Thomas M, Klibanov AM (2003) Conjugation to gold nanoparticles enhances polyethylenimine’s transfer of plasmid DNA into mammalian cells. Proc Natl Acad Sci U S A 100:9138–9143CrossRefGoogle Scholar
  50. 50.
    Zhang G, Yang Z, Lu W, Zhang R, Huang Q, Tian M, Li L, Liang D, Li C (2009) Influence of anchoring ligands and particle size on the colloidal stability and in vivo biodistribution of polyethylene glycol-coated gold nanoparticles in tumor-xenografted mice. Biomaterials 30:1928–1936CrossRefGoogle Scholar
  51. 51.
    Bhattacharya R, Patra CR, Earl A, Wang S, Katarya A, Lu L, Kizhakkedathu JN, Yaszemski MJ, Greipp PR, Mukhopadhyay D (2007) Attaching folic acid on gold nanoparticles using noncovalent interaction via different polyethylene glycol backbones and targeting of cancer cells. Nanomed Nanotechnol Biol Med 3:224–238CrossRefGoogle Scholar
  52. 52.
    Garg S, De A, Nandi T, Mozumdar S (2013) Synthesis of a smart gold nano‐vehicle for liver specific drug delivery. AAPS PharmSciTech 14:1219–1226CrossRefGoogle Scholar
  53. 53.
    Garg S (2013) Development of nano-particulate systems for their applications in biomedical area. Ph.D. Thesis submitted at the Department of Chemistry, University of DelhiGoogle Scholar
  54. 54.
    Kobayashi K, Sumitomo H, Ina Y (1985) Synthesis and functions of polystyrene derivatives having pendant oligosaccharides. Polym J 17:567–575CrossRefGoogle Scholar
  55. 55.
    Besson T, Coudert G, Guillaumet G (1991) Synthesis and fluorescent properties of some heterobifunctional and rigidized 7‐aminocoumarins. J Heterocycl Chem 28:1517–1523CrossRefGoogle Scholar
  56. 56.
    Furniss PS, Hannaford AJ, Smith PWG, Tatchell AR (2006) Vogel’s textbook of practical organic chemistry, 5th edn. Pearson Education, New DelhiGoogle Scholar
  57. 57.
    Stefanko MJ, Gun’ko YK, Rai DK, Evans P (2008) Synthesis of functionalised polyethylene glycol derivatives of naproxen for biomedical applications. Tetrahedron 64:10132–10139CrossRefGoogle Scholar
  58. 58.
    Ladd DL, Henrichs PM (1998) Synthesis and NMR characterization of monomethoxypoly(ethylene glycol) aldehydes from monomethoxypoly(ethylene glycol) tosylates. Synth Commun 28:4143–4149CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Arnab De
    • 1
  • Sushil Mishra
    • 2
  • Seema Garg
    • 2
  • Subho Mozumdar
    • 2
  1. 1.Department of Microbiology and ImmunologyColumbia UniversityNew YorkUSA
  2. 2.Department of ChemistryUniversity of DelhiDelhiIndia

Personalised recommendations