Technology in Cancer Research & Treatment

ISSN 1533-0346
Volume 4, Number 4, August (2005)
©Adenine Press (2005)

Nanotechnology-based Drug Delivery for Cancer K. K. Jain, M.D.

www.tcrt.org Jain PharmaBiotech

Nanobiotechnologies have been applied to improve drug delivery and to overcome some of Blaesiring 7
the problems of drug delivery in cancer. These can be classified into many categories that CH-4057 Basel, Switzerland
include use of various nanoparticles, nanoencapsulation, targeted delivery to tumors of var-
ious organs, and combination with other methods of treatment of cancer such as radiother-
apy. Nanoparticles are also used for gene therapy for cancer. Some of the technologies
enable combination of diagnostics with therapeutics which will be important for the person-
alized management of cancer. Some of the limitations of these technologies and prospects
for future development are discussed.

Keywords: Cancer; Targeted drug delivery; Nanoparticles; Nanotechnology; Nanobiotech-

nology; Anticancer drugs; Cancer therapy; and Gene therapy.

Introduction

Nanotechnology is the creation and utilization of materials, devices, and systems

through the control of matter on the nanometer-length scale, i.e., at the level of
atoms, molecules, and supramolecular structures. This term is applied to
describe the construction and utilization of functional structures with at least one
characteristic dimension measured in nanometers (a nanometer is one billionth
of a meter (10-9 m). Nanobiotechnology is the application of nanotechnology in
life sciences. These technologies are described in detail elsewhere (1). The most
important applications in healthcare are in diagnostics, drug delivery, and devel-
opment of nanomedicine including nanosurgical procedures. Trend towards
miniaturization of carrier particles, i.e., use of microparticles, had already start-
ed prior to the introduction of nanotechnology in drug delivery. These technolo-
gies have been applied to improve drug delivery and to overcome some of the
problems of drug delivery in cancer. Drug delivery in cancer is important for
optimizing the effect of drugs and reducing toxic side effects. Several
nanobiotechnologies, mostly based on nanoparticles, have been used to facilitate
drug delivery in cancer. Although most of the initial research in nanotechnolo-
gy has been done in academic environments, a considerable development for
drug delivery application is now carried out in an industrial environment. Of the
large number of nanoparticle types and other technologies that are available,
those relevant to cancer are classified as shown in Table I.

Nanoparticle-based Drug Delivery in Cancer

Corresponding Author:
Characteristics of nanoparticles used for drug delivery in cancer are listed in K. K. Jain, M.D.
Table II. Email: jain@pharmabiotech.ch

407
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Gold Nanoparticles for Drug Delivery in Cancer
Gold nanoparticles are the most commonly used nanoparti-
cles for diagnostics and drug delivery. The unique chemical
properties of colloidal gold make it a promising targeted
Technology in Cancer Research & Treatment, Volume 4, Number 4, August 2005
Jain
delivery approach for drugs or genes to specific cells. The
physical and chemical properties of colloidal gold permit
more than one protein molecule to bind to a single particle of
colloidal gold. Tumor necrosis factor (TNF) can be bound to
gold nanocrystals and delivered safely and effectively to
tumor-burdened mice and dogs (2). This will form the basis
of a new formulation of the potent anticancer drug TNF-α,
which destroys tumors but it is also very toxic to healthy tis-
sues. By coupling TNF-α to colloidal gold, this will become
a safe as well as effective anticancer therapy.
Gold and silica composite nanoparticles have been investi-
gated as nanobullets for cancer. Gold atoms bind to silicon
atoms and serve as seeds for the growth of aluminum islands.
The large electron affinity of gold causes a significant change
in the electronic structure of silica resulting in a substantial
reduction in the highest occupied and the lowest unoccupied
molecular orbital and the optical gap, thus allowing it to
absorb near infrared radiation. This suggests that a small
cluster can have a similar effect in the treatment of cancer as
the large size nanoshell but with a different mechanism (3).
Dendrimers for Anticancer Drug Delivery
Earlier studies of dendrimers in drug delivery systems
focused on their use for encapsulating drug molecules.
However, it was difficult to control the release of the drug.
One solution to this problem involves the use of dendrimers
with pH-sensitive hydrophobic acetal groups on the den-
drimer periphery. Loss of acetal group at mildly acidic pH
triggers the disruption of micelles and release of the drug.
Dendrimers have been used to facilitate boron neutron cap-
ture therapy as well as photodynamic therapy of cancer.
Nanotechnology-based Drug Delivery for Cancer
Developments in polymer and dendrimer chemistry have pro-
vided a new class of molecules called ‘dendronized polymer’,
i.e., linear polymer that bear dendrons at each repeat unit.
Their behavior differs from that of linear polymers and pro-
vides drug delivery advantages because of their longer circu-
lation time and numerous possibilities peripheral attachments
of drugs (4). Another approach is to attach the drug to the
periphery of the dendrimer so that the release of the drug can
be controlled by incorporating a degradable linkage between
the drug and the dendrimer. DNA-linked dendrimer nan-
ocluster platform enables the delivery of drugs, genetic mate-
rials, and imaging agents to cancer cells, offering the poten-
tial for developing combinatorial therapeutics (5).
In further research at the Michigan Nanotechnology Institute
for Medicine and the Biological Sciences (Ann Arbor, MI),
modified PAMAM dendritic polymers <5 nm in diameter
have been used as carriers (6). They were conjugated to folic
acid as a targeting agent and then coupled to methotrexate
and injected intravenously into animals bearing tumor that
overexpress the folate receptor. Folate molecules bind to
receptors on tumor cell membranes and facilitate the trans-
port of methotrexate to inside of the tumor cell. A computer
model of the dendrimer is shown in Figure 1.
Drug Nanoparticle-albumin Conjugate
Abraxane (Abraxane Oncology), containing paclitaxel as
albumin-bound particles in an injectable suspension, is
approved for the treatment of breast cancer after failure of
combination chemotherapy for metastatic disease or relapse
within 6 months of adjuvant chemotherapy. It is based on
nanoparticle technology, which integrates biocompatible
proteins with drugs to create the nanoparticle form of the
drug having a size of about 100-200 nanometers to overcome
insolubility problems encountered with paclitaxel. The sol-
vent Cremophor- EL, used previously in formulations of
paclitaxel, causes severe hypersensitivity reactions. To
reduce the risk of allergic reactions when receiving paclitax-
el, patients must undergo premedication using steroids and
antihistamines and be given the drug using slow infusions
lasting a few hours. With Abraxane, the active component
(paclitaxel) can be delivered into the body at a 50% higher
dose over 30 minutes. Because Abraxane is solvent-free,
solvent-related toxicities are eliminated. In a randomized
Phase III trial, the response rate of Abraxane was almost
twice that of the solvent-containing drug Taxol (7).
Abraxane enables higher doses of paclitaxel to be adminis-
tered, which may account in part for its increased antitumor
activity. In addition, albumin normally transports nutrients
to cells and has been shown to accumulate in rapidly grow-
ing tumors. Therefore, Abraxane’s increased effectiveness
may also be due to preferential delivery of albumin-bound
paclitaxel to cancer cells. Abraxane is being developed for
Technology in Cancer Research & Treatment, Volume 4, Number 4, August 2005
409
Methotrexate
Folic acid (drug)
(targeting)
G5-
dendrimer
Fluoresceine (platform)
(imaging)
Figure 1: This simplified computer model shows the dendrimer’s branch-
ing structure and how molecules and drugs are attached. Photo credit: Dr.
Jolanta Kukowska-Latallo, Ph.D. and James R. Baker, Jr., M.D. of the
Michigan Nanotechnology Institute for Medicine and Biological Sciences
(Ann Arbor, MI). Reproduced by permission of the original authors.
the treatment of a variety of cancers. In addition to the stan-
dard infusion formulation of Abraxane, oral and pulmonary
delivery formulations are also being investigated.
Anticancer Drug Particles Incorporated in Liposomes
Several injectable and biodegradable systems have been syn-
thesized based on incorporation of anti-estrogens (AEs) in
nanoparticles and liposomes. Both nanospheres and
nanocapsules (polymers with an oily core in which AEs were
solubilized) incorporated high amounts of 4-hydroxy-tamox-
ifen (4-HT) or RU 58668 (8). Liposomes containing various
ratios of lipids enhanced the apoptotic activity of RU 58668
in several multiple myeloma cell lines tested by flow cytom-
etry. These cell lines expressed both estrogen receptor alpha
and beta subtypes. RU-loaded liposomes, administered
intravenously in an animal model induced the arrest of tumor
growth. Thus, the drug delivery of anti-estrogens enhances
their ability to arrest the growth of tumors which express
estrogen receptors and are of particular interest for estrogen-
dependent breast cancer treatment. In addition it represents
a new potent therapeutic approach for multiple myeloma.
Nanoparticles Containing Antisense Oligonucleotides
Antisense oligonucleotides present many problems in drug
delivery. Nanoparticles consisting of human serum albumin
and containing different antisense oligonucleotides have
been used for facilitating drug delivery to tumors (9). The
glutaraldehyde crosslinking procedure of the particle matrix
410
was identified as a crucial parameter for biodegradability and
drug release of the nanoparticles. The drug loading efficien-
cy increased with longer chain length and employment of a
phosphorothioate backbone of antisense compounds. The
entrapment of a fluorescent labeled oligonucleotide within
the particle matrix was used for the detection of the intracel-
lular drug release of the carrier systems. Confocal laser
scanning microscopy revealed that nanoparticles crosslinked
with low amounts of glutaraldehyde, rapidly degraded intra-
cellularly, leading to a significant accumulation of the ASO
in cytosolic compartments of the tumor cells.
Nanoencapsulation of Anticancer Drugs
Enclosing Drugs in Lipid Nanocapsules
The development of less toxic, liposomal formulations of cis-
platin has been hampered by the low water solubility and low
lipophilicity of cisplatin, resulting in very low encapsulation
efficiencies. A novel method enables the efficient encapsula-
tion of cisplatin in a lipid formulation; it is based on repeated
freezing and thawing of a concentrated solution of cisplatin in
the presence of negatively charged phospholipids (10). The
method is unique in that it generates nanocapsules, which are
small aggregates of cisplatin covered by a single lipid bilay-
er. The nanocapsules have an unprecedented drug-to-lipid
ratio and an in vitro cytotoxicity up to 1000-fold higher than
the free drug. Analysis of the mechanism of nanocapsule for-
mation suggests that the method may be generalized to other
drugs showing low water solubility and lipophilicity.
SuperFluids™ technology of Aphios Corporation (Woburn,
MA) uses phospholipid nanosomes to form stable biocom-
patible aqueous formulations of poorly soluble anticancer
and antiviral drugs such as paclitaxel and camptothecin for
intravenous administration. The process has been used for
the nanoencapsulation of paclitaxel in a formulation called
Taxosomes™, which reduces the toxicity and doubles the
efficacy of the drug as shown in studies using nude mice
with breast cancer xenografts. The process has also been
used for the nanoencapsulation of camptothecin in a stable
aqueous liposomal formulation called Camposomes™.
Water soluble derivatives of camptothecin, a unique topoiso-
merase 1 inhibitor, have been approved by the FDA for use
in colorectal cancer. Camposomes™ have been shown to be
very effective against lymphomas in nude mice.
Encapsulating Drugs in Hydrogel Nanoparticles
NOF Corporation (Tokyo, Japan) has developed Hydrogel-
Nanoparticles based upon hydrophobic polysaccharides
(PUREBRIGHT®) for drug delivery in which proteins
and/or antibodies can be encapsulated. Cholesterol pullulan
shows unique characteristics. In the water four cholesterol
Technology in Cancer Research & Treatment, Volume 4, Number 4, August 2005
Jain
molecules gather to form hydrophobic core, then usually
eleven units of these core make polymers self-aggregated
with localizing pullulan outside. Resulting cholesterol
nanoparticles can stabilize proteins and/or antibodies by
forming hybrid-complex. These particles also stimulate
immune system as they are easily trapped by dendritic cells
in the blood, so that vaccine therapy for cancer may be facil-
itated by encapsulating cancer specific MAbs such as Her2.
Exosomes for Cancer Drug Delivery
Exosomes are small (50-100 nm), spherical vesicles pro-
duced and released by most cells to facilitate intercellular
communication. These vesicles are of endosomal origin
and are secreted in the extracellular milieu following fusion
of late endosomal multivesicular bodies with the plasma
membrane. They have a defined protein composition that
confers specific biological activities contingent on the
nature of the producing cell. Tumors release membranous
material mimicking these ‘exosomes,’ resulting in deletion
of reactive lymphocytes. Although exosomes express tumor
antigens, leading to their proposed utility as tumor vaccines,
they also can suppress T-cell signaling molecules and
induce apoptosis (11). The first phase I clinical trial using
autologous exosomes pulsed with MAGE 3 peptides for the
immunization of stage III/IV melanoma patients has shown
the feasibility of large scale exosome production and the
safety of exosome administration (12).
Exosomes produced by dendritic cells are called dexosomes
and contain essential components to activate both adaptive
and innate immune responses. Anosys (Menlo Park, CA) is
developing dexosome vaccines that use patient-specific dex-
osomes loaded with tumor antigen-derived peptides to treat
cancer. Exosome research continues to reveal unique prop-
erties which broaden their fields of application. Anosys’
Exosome Display Technology provides the ability to manip-
ulate exosome composition and tailor exosomes with new
desirable properties opening up opportunities in the field of
recombinant vaccine and monoclonal antibody (MAb)
preparation. This is achieved by generating genes coding for
chimeric proteins linking an exosome addressing sequence
to antigens or biologically active proteins. The resulting pro-
teins are targeted to exosomal compartment and released in
the extracellular milieu bound to exosomes.
Targeted Drug Delivery with Nanoparticles
PEG-coated Nanoparticles
PEG-coated biodegradable nanoparticles can be coupled
to folic acid to target the folate-binding protein; this mol-
ecule is the soluble form of the folate receptor that is over-
expressed on the surface of many tumor cells. The specif-
Nanotechnology-based Drug Delivery for Cancer
ic interaction between the conjugate folate-nanoparticles
and the folate-binding protein has been evaluated by sur-
face plasmon resonance and confirmed a specific binding
of the folate-nanoparticles to the folate-binding protein
(13). Thus, folate-linked nanoparticles represent a poten-
tial new drug carrier for tumor cell-selective targeting.
PEG-coated biodegradable polycyanoacrylate nanoparti-
cles conjugated to transferrin could also be an effective
carrier for paclitaxel delivery (14).
Carbon Magnetic Nanoparticles for Targeted Drug Delivery
in Cancer
Carbon magnetic nanoparticles (CMNP) are composed of
spherical particles, 40-50 nm in diameter, with iron/iron
oxide particles dispersed in a carbon-based host-structure
(15). Free doxorubicin (DOX) molecules are immobilized
onto the surfaces of activated CMNP particles to form
CMNP-DOX conjugates. The in vitro antiproliferative
activity of immobilized doxorubicin in the conjugates has
been demonstrated in tumor cell cytotoxicity assays. It is
suggested that this CMNP-DOX system can be used for tar-
geted drug-delivery systems in cancer.
Targeted Drug Delivery with
Nanoparticle-aptamer Bioconjugates
Nucleic acid ligands (aptamers) are potentially well suited
for the therapeutic targeting of drug encapsulated controlled
release polymer particles in a cell- or tissue-specific manner.
Poly(lactic acid)-block-polyethylene glycol (PLA-PEG)
copolymer have been synthesized with a terminal carboxylic
acid functional group (PLA-PEG-COOH), and encapsulated
rhodamine-labeled dextran (as a model drug) within PLA-
PEG-COOH nanoparticles (16). These nanoparticles have
the following desirable characteristics:
• Negative surface charge may minimize nonspe-
cific interaction with the negatively charged
nucleic acid aptamers.
• Carboxylic acid groups on the particle surface
facilitates covalent conjugation to amine-modi-
fied aptamers.
• Presence of PEG on particle surface decreased
uptake in nontargeted cells and enhances circu-
lating half-life.
Nanoparticle-aptamer bioconjugates, generated with RNA
aptamers, bind to the prostate-specific membrane antigen
(PSMA), a well-known prostate cancer tumor marker that
is overexpressed on prostate acinar epithelial cells. These
bioconjugates could efficiently target and get taken up
selectively by the prostate epithelial cells, which express
the PSMA protein.
Technology in Cancer Research & Treatment, Volume 4, Number 4, August 2005
411
Nanodroplets for Site-specific Cancer Treatment
Nanodroplets are being developed by ImaRx Therapeutics
(Tucson, AZ) for site-specific cancer treatment. Nano-
droplets will be loaded with cancer drugs and targeted to
specifically seek out cancer cells and release the drug pay-
load. The therapy will also use ultrasound to further aid in
specifically targeting cancer cells. One purpose of this
research is to develop a nanoparticle delivery system for
camptothecin-based drugs because their poor solubility and
labile lactone ring pose challenges for drug delivery. After
initial investigations SN-38 was selected as the candidate
camptotheca alkaloid for further development.
Nanoparticles comprising SN-38, phospholipids and poly-
ethylene glycol were developed and studied in vitro and in
vivo (17). The SN-38 formulations were stable in human
serum albumin and high lactone concentrations were
observed even after 3 h. In vivo studies in nude mice
showed prolonged half-life of the active (lactone form) drug
in whole blood and increased efficacy compared to
Camptosar in a mouse xenograft tumor model.
Lipid-based Nanocarriers
LiPlasomes, which are in development by LiPlasome
Pharma (Lyngby, Denmark), are smart lipid-based
nanocarriers that can be applied for targeted transport of
anticancer drugs. The targeted drug delivery principle
consists of long circulating nanoparticles such as lipo-
somes or micelles that accumulate in porous cancer tissue
with a high phospholipase A 2 (PLA2) activity. The carri-
er nanoparticles are composed of special prodrug lipids
whose degradation products, after exposure to PLA2, are
converted to active drugs such as anticancer lysolipids
and/or fatty acid drug derivatives (18). The PLA2 hydrol-
ysis products will furthermore act as locally generated
permeability enhancers that promote the absorption of the
released drugs across the cancer cell membranes into puta-
tive intracellular target sites. This innovative prodrug and
drug delivery concept allows for intravenous transport of
high concentrations of anticancer drugs directly to the
tumor target. It enables, without any prior knowledge of
the position and size of the tumor, to release the anticancer
drugs specifically at the tumor target site.
The Polymerized Liposomal Nanoparticle (PLN) of
NanoMed Technologies LLC (Columbus, OH) is a non-
viral nanoparticle technology incorporating a targetable
and customizable drug delivery system for chemothera-
peutic applications. The nanoparticles are non-immuno-
genic, display no acute toxicity and can be highly concen-
trated. Intracellular degradation and excretion rates of the
particles can be modulated by controlling the degree of
polymerization.
412
Targeted Antiangiogenic Therapy using Nanoparticles
Integrin-targeted nanoparticles for site-specific delivery of a
therapeutic payload form the basis of proprietary technolo-
gy of Targesome (Palo Alto, CA). Selective targeting of
upregulated αvβ3 and Flk-1 on the neovasculature of
tumors is a novel antiangiogenesis strategy for treating a
wide variety of solid tumors. A study provides proof of
principle that targeted radiotherapy works using different
targeting agents on a nanoparticle (NP), to target both the
integrin αvβ3 and the vascular endothelial growth factor
receptor (19). These encouraging results demonstrate the
potential therapeutic efficacy of the IA-NP-90Y and anti-
Flk-1 MAb-NP-90Y complexes as novel therapeutic agents
for the treatment of a variety of tumor types.
The synthetic peptide bearing Arg-Gly-Asp (RGD) sequence
is considered to specifically bind to αvβ3 integrin expressed
on endothelial cells in the angiogenic blood vessels, which
provides a potential to inhibit the tumor growth. Hydropho-
bically modified glycol chitosan, capable of forming nano-
sized self-aggregates, was used as a carrier for the RGD pep-
tide, which was labeled with fluoresein isothiocyanate
(FITC-GRGDS) and loaded into self-aggregates by solvent
evaporation methods (20). The self-aggregates loaded with
FITC-GRGDS might be useful for monitoring or destroying
the angiogenic vessels surrounding the tumor tissue.
Nanoparticles for Delivery of Drugs to Brain Tumors
Nanoparticles may be especially helpful for the treatment of
the disseminated and very aggressive brain tumors.
Intravenously injected doxorubicin-loaded polysorbate 80-
coated nanoparticles enabled a 40% cure rate in rats with
intracranially transplanted glioblastomas 101/8 (21). A fur-
ther study has evaluated the acute toxicity of doxorubicin
associated with polysorbate 80-coated nanoparticles in
healthy rats and to establish a therapeutic dose range for this
formulation in rats with intracranially implanted 101/8
glioblastoma (22). The presence of polysorbate 80 in the for-
mulations was not associated with changes in toxicity com-
pared with free or nanoparticulate drug. The results in tumor-
bearing rats were similar to those in healthy rats. These results
demonstrate that the cardiotoxicity of the doxorubicin bound
to nano-particles was lower than that of free doxorubicin.
Smart superparamagnetic iron oxide particle conjugates can
be used to locate brain tumors earlier and more accurately
than current methods and to target the tumors (23). Use of
folic acid com-bined with PEG can further enhance the spe-
cific targeting capability of the nanoparticles and enhance
their intracellular uptake. A variety of small molecules tar-
geting tumor receptors and even chemotherapy agents can be
attached to the nanoparticles.
Technology in Cancer Research & Treatment, Volume 4, Number 4, August 2005
Jain
Doxorubicin, an anticancer drug, bound to polysorbate-coat-
ed nanoparticles crosses the intact blood-brain barrier and
reaches therapeutic concentrations in the brain. NanoDel
technology has been shown to deliver doxorubicin to brain
tumors. Another formulation of doxorubicin with polysor-
bate-coated nanoparticles has been administered in rat mod-
els with implanted brain tumors (24). Rats treated with dox-
orubicin bound to polysorbate-coated nanoparticles had sig-
nificantly higher survival times compared with all other
groups. Over 20% of the animals in this group showed a
long-term remission. This study showed that therapy with
doxorubicin bound to nanoparticles offers a therapeutic
potential for the treatment of human glioblastoma.
Combination of Nanoparticles with Radiotherapy
Application of Dendrimers in Boron Neutron Capture Therapy
Boron neutron capture therapy (BNCT) offers a potential
method for localized destruction of tumor cells. The technolo-
gy is based on the nuclear reaction between thermal neutrons
and boron-10 (10B) to yield alpha particles and lithium-7
nuclei. The destructive effect of this reaction is limited to a
range of about the diameter of a single cell. In order for BNCT
to be effective in cancer therapy, there must be selective deliv-
ery of an adequate concentration of 10B to tumors. Various
types of antibodies as well as epidermal growth factor have
been utilized to investigate receptor-mediated boron delivery,
however in vivo studies have demonstrated only a small per-
centage of the total administered dose actually accumulates in
tumors while high concentrations end up in the liver.
In normal as well as cancer cells, the low molecular weight
vitamin, folic acid, is required for a number of enzymatic
pathways. Cell membrane receptors mediating endocytic
transport of folic acid into cells are expressed in elevated lev-
els in a variety of human tumors. Folic acid conjugates with
macromoledules such as toxins, enzymes, antibodies, genes,
and liposomes have been shown to be internalized into tumor
cells overexpressing folate receptors. These strategies have
been employed to enhance the effect of BNCT. The use of
dendrimers as boron carriers for antibody conjugation is
based on their well-defined structure and multivalency.
The use of dendrimers as boron carriers for antibody conjuga-
tion is based on their well-defined structure and multivalency.
Boronated PAMAM dendrimers have been designed to target
the epidermal growth factor receptor, a cell surface receptor
that is frequently overexpressed in brain tumor cells (25).
Nanoengineered Silicon for Brachytherapy
BrachySil™, a nanoparticle preparation containing immobi-
lized isotope 32-phosphorus, is in development as
Nanotechnology-based Drug Delivery for Cancer
brachytherapy by pSiMedica Ltd (Worcs, UK). It demon-
strates a very high degree of isotope retention following
injection into the liver, thus reducing the risk of soluble
radioactive material affecting healthy hepatic tissue, or
entering the circulation and causing systemic toxicity.
Unlike titanium seeds, which remain for ever in the body,
phosphorus seeds degrade over time and enable repetition of
treatment if necessary. Other treatments for primary liver
cancer include a variety of embolization and radio-frequen-
cy ablation techniques. BrachySil offers a more versatile
and safer product for the treatment of such tumors. The pro-
cedure is undertaken without surgery under local anesthetic
and patients can be discharged the following day. A phase
IIa trial in primary liver cancer has shown that it is safe and
effective in tumor regression with increased efficacy.
Following the completion of analysis of the final Phase IIa
trial results, a dose profiling study will be conducted during
2005 followed by multicenter pivotal registration trials to
obtain data to support registration of BrachySil as an
approved treatment for primary liver cancer. Its use will be
expanded for a wider range of solid tumor indications.
Combination of Nanoparticles with Physical Modalities of
Cancer Therapy
Nanoshells and Laser Ablation of Tumors
Nanospectra Biosciences Inc (Houston, TX) is developing
nanoshells for the targeted destruction of various cancers.
The company’s research to date has demonstrated its ability
to target specific cells or tissues and non-invasively ablate
tumors using an external laser without damage to surround-
ing tissue. The blood vessels inside tumors develop poorly,
allowing small particles like nanoshells to leak out and
accumulate inside tumors. The advantages of Nanoshell-
based tumor cell ablation include:
• Targeting to specific cells and tissues to avoid
damage to surrounding tissue;
• Less adverse effects than targeted chemothera-
peutic agents or photodynamic therapy;
• Repeatability because of lack of “tissue memory”
as in radiation therapy; and
• Ability to treat such as glioblastoma multiforme,
metastases and inoperable tumors.
Nanotechnology-based Adjuncts to Photodynamic Therapy
Photodynamic therapy (PDT) uses light-activated drugs
called photosensitizers to treat a range of diseases character-
ized by rapidly growing tissue, including the formation of
abnormal blood vessels, such as cancer. The more tradition-
al name for this therapy is photoradiation therapy. Treatment
with PDT consists of a two-step process that starts with
Technology in Cancer Research & Treatment, Volume 4, Number 4, August 2005
413
administration of the drug, or photosensitizer, by intravenous
injection. Once the drug enters the bloodstream, it attaches
itself to low-density lipoproteins already circulating. As cells
undergoing rapid growth require an above-average supply of
lipoproteins, the drug reaches these types of cells more quick-
ly and in higher concentrations. Once the necessary level of
concentration is attained, the second step is to activate the
drug with a specific dose of light of a particular wavelength.
This causes the conversion of normal oxygen found in tissue
to a highly energized form called singlet oxygen, which in
turn, disrupts normal cellular functions. Neither the drug nor
the light exerts any effect until combined.
Numerous studies have used liposomes, oils, and polymer-
ic micelles as encapsulation methods, with some success.
However, all of these techniques suffer from one unpleas-
ant side effect: after controlled release and photosensitiza-
tion, the drug is free to circulate the body, accumulating in
the eyes and skin. This leads to phototoxic side effects,
rendering the patient highly sensitive to light. A further
disadvantage is that liposomes can be engulfed and
destroyed by cells of the RES. Such problems have limit-
ed the emerging field of PDT, but combination of this tech-
nique with nanotechnology is promising.
The possibility of improving dendrimers through appropriate
functionalization of their periphery makes them promising
carriers of PDT. The use of 5-aminolevulinic acid (ALA) is
one approach to PDT based on dendrimers. ALA is a natural
precursor of the photosensitizer protoporphyrin IX (PIX) and
its administration increases the cellular concentrations of PIX.
An approach to deep tissue penetration is based on two-pho-
ton excitation with near-infrared lasers. Multivalent aspects
of dendrimer scaffold have been used to conjugate several
two-photon absorbing chromophores to the porphyrin core
(26). This system has been shown to generate singlet oxy-
gen efficiently on light irradiation at 780 nm wavelength.
Dynamic ceramics is a new development that addresses the
various problems of PDT for cancer. An adjunct anticancer
drug, 2-devinyl-2-(1-hexyloxyethyl) pyropheophorbide
(HPPH), was encapsulated within a ceramic-based nanopar-
ticle, which are ∼35 nm diameter and are made from silica
(27). They are stable to fluctuations in temperature and pH
and small enough to evade the reticuloendothelial system
(RES). The success of the technique relies upon the tiny
pores in the ceramic particle, which range from 0.5-1.0 nm.
These are too small to allow the drug to escape its encapsu-
lation but small enough to enable oxygen to diffuse back and
forth. Thus, HPPH can exert tumor-killing effects without
being released into the blood stream. The ceramic nanopar-
ticles accumulate exclusively in the tumor tissue without the
need for active targeting. After phototherapy, the ceramic
414
spheres retain the drug, and are unlikely to cause the side
effects that were seen with earlier encapsulation methods. In
an extension of the technique, a magnetic core is encapsulat-
ed within a silica nanoparticle and targeted to tumor tissue.
The magnetically impregnated tumor cells could then be
killed by exposing the tissue to a magnetic field, which is a
potentially complementary technique to PDT.
Iron Nanoparticles and Thermal Ablation of Tumors
An experimental procedure for the treatment of breast can-
cer called “magnetic thermal ablation” has been examined
under in vivo animal conditions. The method consists in the
intratumoral application of iron nanoparticles and the expo-
sure of the breast to an alternating magnetic field, whereby
the tumor is eliminated by heat (28). An innovation of ther-
mal ablation developed by Triton BioSystems (Chelmsford,
MA) is a non-invasive method of killing cancer using local-
ized lethal heat with negligible damage to healthy tissues.
This approach bonds iron nanoparticles and MAbs into bio-
probes. The Company’s product is referred to as the
Targeted Nano-Therapeutics™ (TNT) system. The TNT
system consists of two components:
I. The injectable component referred to as T-
probes. Each T-probe is made of two parts: (i) a
MAb that acts only as a guidance system direct-
ing the T-probe (40 nanometers long) to cancer
cells and (ii) a nanoscale particle made of a spe-
cial material composite, that constitutes the
lethal payload when activated.
II. The Magnetic Field Device that serves to activate
the T-probes in the treatment area.
Trillions of T-probes are dispensed into the body in serum
form by infusion into the blood stream. Once the T-probes
are attached to cancer cells, a focused magnetic field selec-
tively activates their magnetic particles. The magnetic field
energy is converted to lethal heat by the particles causing a
rapid temperature increase to more than 1700° C at the sur-
face of the cancer cells, killing them and their blood supply
with negligible damage to surrounding healthy tissues. The
Company believes this will eliminate many of the side effects
currently associated with chemotherapies and radiotherapies.
Anti-HER2 antibody can induce antitumor responses, and
can be used in delivering drugs to HER2-overexpressing
cancer. Anti-HER2 immunoliposomes containing mag-
netite nanoparticles, which act as tumor-targeting vehi-
cles, have been used to combine anti-HER2 antibody ther-
apy with hyperthermia (29). When introduced into
SKBr3 breast cancer cells in vitro, 60% of magnetite
nanoparticles incorporated into SKBr3. The cells were
then heated at 42.5° C under an alternating magnetic field,
Technology in Cancer Research & Treatment, Volume 4, Number 4, August 2005
Jain
resulting in strong cytotoxic effects. These results sug-
gest that this novel therapeutic tool is applicable to treat-
ment of HER2-overexpressing cancer.
Interaction of Nanoparticles with Ultrasound Radiation
Nanoparticles have been introduced in tumors followed by
ultrasound-induced cavitation for safe and efficient drug and
gene delivery. In a study on athymic nude mice bearing
human colon KM20 tumors, polystyrene nanoparticles (100-
280 nm) were injected intravenously in combination with
ultrasound to enhance delivery of chemotherapeutic agent 5-
fluorouracil (30). This combination significantly decreased
tumor volume and resulted in complete tumor regression at
optimal irradiation conditions.
Nanoparticle-mediated Gene Therapy
The success of the gene therapy for clinical applications, in
part, would depend on the efficiency of the expression vec-
tor as determined by the level as well as the duration of
gene expression. Although various cationic polymers and
lipid-based systems are being investigated, most of these
systems exhibit higher-level but transient gene expression.
Most often, the emphasis is on the level of gene expression
rather than on the duration of gene expression. In certain
disease conditions, a relatively low level of gene expres-
sion (therapeutic level) but for a sustained duration may be
more effective than higher-level but transient gene expres-
sion. Therefore, a gene expression system that can modu-
late the level as well as the duration of gene expression in
the target tissue is desirable. Polymer-based sustained
release formulations such as nanoparticles have the poten-
tial of developing into such a system.
Nanoparticles for p53 Gene Therapy of Cancer
Transfer of p53 gene with sustained expression of the p53
protein in the target cells is a recognized approach in can-
cer gene therapy. A single-dose regimen results in only a
weak and transient inhibition of cell proliferation. Gene
delivery with nanoparticles usually requires direct intratu-
moral injection but tumor targeting via intravascular
administration is possible if nanoparticle surface is modi-
fied to avoid trapping of the nanoparticles by the reticu-
loendothelial system. Another approach is the use of
poly(D,L-lactide-co-glycolide) nanoparticles loaded with
wild type (wt) p53 DNA, which has demonstrated a sus-
tained antiproliferative effect in a breast cancer cell line
following slow intracellular release of the encapsulated
DNA from nanoparticles (31). The results of the study
suggest that wt-p53 DNA-loaded nanoparticles have
potential use in the therapy of breast and other cancers
associated with mutations in the p53 gene.
Nanotechnology-based Drug Delivery for Cancer
Intravenous Nanoparticle Formulation for
Delivery of FUS1 Gene
INGN 401 from Introgen Therapeutics Inc (Austin, TX) is an
intravenous nanoparticle formulation (liposomal
DOTAP:Chol-FUS1 complex) of the tumor suppressor gene
FUS1, which suppresses tumor growth and has led to tumor
regression in mouse models of metastatic lung cancer (32).
Mice treated with INGN 401 survived almost 70% longer than
untreated mice. Phase I clinical trials in patients with lung
cancer have started and initial survival results are encourag-
ing. Combining the systemic delivery capabilities of the
nanoparticle delivery with the cancer-specific activity of
tumor suppressor genes, such as FUS1, may provide a new
approach to meet the challenge of treating metastases as it has
the ability to attack cancer at multiple sites throughout the
body. Systemic administration of DNA-nanoparticles howev-
er, is associated with inflammation, which can be suppressed
by small molecule inhibitors without affecting transgene
expression (33). This strategy may be incorporated into future
clinical trials based on systemic non-viral vector gene therapy.
Strategies Combining Diagnostics and Therapeutics
Nanoshells as Adjuncts to Thermal Tumor Ablation
Metal nanoshells belong to a class of nanoparticles with tun-
able optical resonances that have been used for thermal abla-
tive therapy for cancer. Nanoshells can be tuned to strongly
absorb light in the near infrared range, where optical trans-
mission through tissue is optimal. Nanoshells placed at depth
in tissues can be used to deliver a therapeutic dose of heat by
using moderately low exposures of extracorporeally applied
near-infrared. In vivo studies under magnetic resonance guid-
ance have revealed that exposure to low doses of near-
infrared light in solid tumors treated with metal nanoshells
reached average maximum temperatures capable of inducing
irreversible tumor destruction within 4-6 min (34).
The ability to control both wavelength-dependent scattering
and absorption of nanoshells offers the opportunity to design
nanoshells which provide both diagnostic and therapeutic
capabilities in a single nanoparticle. A nanoshell-based all-
optical platform technology can integrate cancer imaging
and therapy applications. Immunotargeted nanoshells, engi-
neered to both scatter light in the near infrared range
enabling optical molecular cancer imaging and to absorb
light, enable selective destruction of targeted carcinoma cells
through photothermal therapy (35). In a proof of principle
experiment, dual imaging/therapy immunotargeted nano-
shells were used to detect and destroy breast carcinoma cells
that overexpress HER2, a clinically relevant cancer bio-
marker. The new approach has some significant advantages
over other alternatives that are under development. For
Technology in Cancer Research & Treatment, Volume 4, Number 4, August 2005
415
example, optical imaging is much faster and less expensive
than other medical imaging techniques. Gold nanoparticles
are also more biocompatible than other types of optically
active nanoparticles, such as quantum dots.
Paramagnetic Nanoparticles
Tiny melanoma tumors growing in mice, indiscernible from
the surrounding tissue by direct MRI scan, can be easily
located by MRI scan performed 30 minutes after the mice
are injected with paramagnetic nanoparticles (36).
Nanoparticles were equipped with tiny hooks that link only
to complementary loops found on cells in newly forming
blood vessels of the tumors, revealing their location. One of
the advantages of nanoparticles is that they enable detection
of tumors by using the same MRI equipment that is in stan-
dard use for heart or brain scans, so that the technique is use-
ful in a hospital setting. Another advantage is that the parti-
cles can be loaded with a wide variety of drugs that can be
targeted to growing tumors. The nanoparticles may also
enable more effective follow up, because a physician could
use them to detect whether a tumor was still growing after
radiation or chemotherapy treatments.
Concluding Remarks
Although most of the technologies described are promising
and fit well with the current methods of treatment, there are
still safety concerns of the introduction of nanoparticles in the
human body. These will require further studies before some
of the products can be approved. A few nanobiotechnolgy-
based products have been approved and include the anticancer
drug Abraxane. The most promising methods of drug deliv-
ery in cancer will be those that combine diagnostics with treat-
ment. These will enable personalized management of cancer
and provide an integrated protocol for diagnosis and follow-
up that is so important in management of cancer patients.
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Date Received: May 11, 2005
Date Accepted: June 20, 2005