Published on Web 01/09/2009

Entrapment of Hydrophobic Drugs in Nanoparticle Monolayers with Efficient

Release into Cancer Cells
Chae Kyu Kim,† Partha Ghosh,† Chiara Pagliuca,‡ Zheng-Jiang Zhu,† Stefano Menichetti,‡ and
Vincent M. Rotello*,†
Department of Chemistry, UniVersity of Massachusetts at Amherst, Amherst, Massachusetts 01003, and
Dipartimento di Chimica Organica “U. Schiff”, UniVersità di Firenze, 50019, Italy
Received October 15, 2008; E-mail: [email protected]

Drug delivery systems (DDSs) provide an important tool for

increasing the efficacy of pharmaceuticals through improved
pharmacokinetics and biodistribution.1 A wide variety of nanoscale
materials such as liposomes, polymeric micelles, and dendrimers
have been employed as drug carriers.2 Both covalent and nonco-
valent approaches can be applied to the conjugation of drugs into/
onto these DDSs.3 Noncovalent approaches have the capability of
employing active drugs, whereas covalent attachment generally
requires chemical modification which can cause reduced efficiency
of drug release or incomplete intracellular processing of a prodrug.4
Recently, gold nanoparticle (AuNP) based drug/gene delivery
systems have attracted attention due to their functional versatility,5
biocompatibility,6 and low toxicity.7 Recent studies have demon-
strated a controlled release of payload by intracellular thiols.8
However, controlled dissociation of drugs in active form from
covalent AuNP-drug conjugates remains a challenge for clinical
applications.2
Noncovalent incorporation of drugs into AuNP monolayers
provides an alternative delivery strategy with the potential for
avoiding drug release and prodrug processing issues. The structure
of commonly used water-soluble AuNPs is similar to that of
unimolecular micelles such as dendrimers, featuring a hydrophobic
interior and a hydrophilic exterior.9 The alkanethiol monolayer of
the nanoparticle coupled with the radial nature of the ligands10
creates “hydrophobic pockets” inside the monolayer of the AuNP
where organic solutes can be partitioned, as demonstrated by
Lucarini and Pasquato.11 We report here the use of these pockets
to encapsulate drugs and deliver them with high efficiency to cells.
The biocompatible AuNPs used in this study feature two
functional domains: a hydrophobic alkanethiol interior and a
hydrophilic shell composed of a tetra(ethylene glycol) (TEG) unit
terminated with a zwitterionic headgroup. Particles with this general
structure have been shown to minimize nonspecific binding with Figure 1. (a) Delivery of payload to cell through monolayer-membrane
interactions. (b) Structure of particles and guest compounds: Bodipy, TAF,
biomacromolecules.12 and LAP, the number of encapsulated guests per particle, and log P of the
We chose three different hydrophobic guest compounds: 4,4- guests. (c) Release of Bodipy from AuNPZwit-Bodipy in DCM-aqueous
difluoro-4-bora-3a,4a-diaza-s-indacene (Bodipy) as a fluorescent solution two-phase systems (λex) 499 nm, λem) 517 nm). (d) PL intensity
probe13 and the highly hydrophobic therapeutics tamoxifen (TAF) of AuNPZwit-Bodipy in cell culture medium and 100% serum, indicating
little or no release relative to AuNPZwit-Bodipy in PBS after NaCN-
and β-lapachone (LAP) as drugs (Figure 1). The nanoparticle-payload induced release of guest molecules (λex) 499 nm, λem) 510 nm).
conjugates (AuNPZwit-Bodipy, TAF, and LAP) were prepared
by a solvent displacement method.14 First, AuNPZwit (Au core: determined from 1H NMR spectra and NaCN-induced decomposi-
2.5 ( 0.4 nm) and guest were dissolved in acetone/water, and the tion experiments (see Supporting Information) and varied depending
solvent slowly evaporated. The bulk of the excess guest precipitated on size, hydrophobicity (log P), and molecular structure of
out and was removed by filtration; the particles were further purified hydrophobic molecules (Figure 1). The particle/guest complexes
by multiple filtrations through a molecular weight cutoff filter until are stable in buffer for >1 month and to extended dialysis, a level
no free guest was observed, followed by dialysis against buffer. of kinetic entrapment greater than that observed with dendrimers.4
The number of entrapped guest molecules per particle was The ability of the delivery systems to release their payload was
first explored in Vitro using AuNPZwit-Bodipy in a two-phase
†
University of Massachusetts at Amherst.
dichloromethane (DCM)-water system,15 where the dye is quenched
‡
Università di Firenze. by the AuNP and photoluminescence (PL) was only observed upon
1360 9 J. AM. CHEM. SOC. 2009, 131, 1360–1361 10.1021/ja808137c CCC: $40.75  2009 American Chemical Society
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Figure 3. Cytotoxicity of AuNPZwit complexes measured by Alamar blue

assay after 24 h incubation with MCF-7 cells. IC50 of AuNP (NP), equivalent
Figure 2. CLSM images of MCF-7 cell treated with AuNPZwit-Bodipy drugs (Drug), and free drugs are shown in table.
for 2 h: (a) green channel, (b) bright field, and (c) overlap. TEM images of
fixed cell treated with (d) AuNPZwit-Bodipy and (e) AuNPTTMA as a accumulation in tumor tissues by the enhanced permeability and
positive control. Endosomally trapped AuNPs are marked by arrow. (f) ICP- retention (EPR) effect.17 Additionally, the noninteracting nature of
MS measurement (200 000 cells/well), indicating low cellular uptake of
AuNPZwit (31 ng/well after 4 h). their monolayer should make these systems highly amenable to
targeting strategies. We are currently exploring these applications
dye release. In these studies, a rapid increase in PL intensity is as well as the role of monolayer and guest structure in the
observed along with transfer of Bodipy into the DCM layer (Figure encapsulation process.
1c). Significantly, since no release is observed in monophasic
aqueous conditions and no particle was observed in the DCM layer, Acknowledgment. This research was support by the NIH
payload release occurs interfacially. (GM077173).
Payload delivery to cells using AuNPZwit-Bodipy was deter- Supporting Information Available: Experimental procedures,
mined by confocal laser scanning microscopy (CLSM) using human synthesis of gold nanoparticles and complexes, and TEM of gold
breast cancer (MCF-7) cells. Efficient delivery of the dye to the nanoparticles. This information is available free of charge via the
cytosol is observed after 2 h of incubation with AuNPZwit-Bodipy Internet at http://pubs.acs.org.
(Figure 2a-c). Cellular uptake of nanoparticles was studied using
transmission electron microscopy (TEM) and inductively coupled References
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tionality should provide long circulation lifetimes and preferential JA808137C

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