Nanovector therapeutics
Mauro Ferrari1,2
An ideal injected therapeutic drug would travel through the
vasculature, reach the intended target at full concentration, and
there act selectively on diseased cells and tissues only, without
creating undesired side effects. Unfortunately, even the best
current therapies fail to attain this ideal behavior, by a wide
margin. A primary reason is the fact that the target recognition
abilities of the current therapeutics molecules are quite limited.
Furthermore, the natural defenses of the body present a
sequence of formidable obstacles on the drug’s pathway to the
intended lesion. Requiring any molecule to have sufficient
therapeutic efficacy, target recognition specificity, as well as all
of the tools required to bypass multiple biological barriers is
probably unrealistic. A different approach is to decouple the
problem (i.e. employ the drug molecules for their therapeutic
action only, and deliver them to the intended site by vectors
that can be preferentially concentrated at desired body
locations through the concurrent action of multiple targeting
mechanisms). These vectors must also be large enough to
comprise all the requirements for the evasion of the body
defenses, while still sufficiently small so as not to create
undesired blockages of even the smallest of blood vessels —
and thus, by definition, nanotechnological.
Addresses
1
The Ohio State University, 473 West 12th Avenue, Columbus OH
43210-1002, USA
2
National Cancer Institute, 31 Center Drive MSC 2580, Room 10A52,
Bethesda, MD 20892, USA
Corresponding author: Ferrari, M (ferrari@lvd1.bme.ohio-state.edu)
Current Opinion in Chemical Biology 2005, 9:343–346
This review comes from a themed issue on
Next-generation therapeutics
Edited by Chris J Vlahos And Michael Coghlan
Available online 20th June 2005
1367-5931/$ – see front matter
# 2005 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.cbpa.2005.06.001
Introduction
Injected, nano-scale drug delivery systems, or ‘nanovec-
tors’, are ideal candidates to provide breakthrough solu-
tions to the time-honored problem of optimizing
therapeutic index for a treatment (i.e. to maximize effi-
cacy, while reducing health-adverse side effects). Even
modest amounts of progress towards this goal have his-
torically engendered substantial benefits across multiple
fields of medicine, with the translability from, for exam-
ple, a subfield of oncology to a field as distant as the
www.sciencedirect.com
treatment of infectious disease being granted by the fact
that the progresses had a single common denominator in
the underlying technological platform. The case of lipo-
somes illustrates the point, in view of the advances they
have yielded in the treatment of breast and ovarian
cancers, Kaposi’s sarcoma, as well as fungal infections
in multiple clinical contexts. Liposomes were the first
nanovector therapy to reach health-care fruition, have
enjoyed widespread clinical use for over 10 years, and
continue to draw substantial research interest. They have
shown the way towards the revolutionary advances that
medicine can expect in the current decade. Today, many
hundreds if not thousands of entirely different nanovector
technology platforms have joined liposomes at this fron-
tier, each with different properties, strengths, and weak-
nesses. Most frequently discussed among these are
polymer-based platforms [1,2,3–5], dendrimers [6–8],
gold nano-shells [9,10–12], semiconductor nano-crystals
[13,14,15], carbon-60 fullerenes, biologically derived
nano-constructs [16–18], silicon- and silica-based nano-
systems [19,20,21] and superparamagnetic nanoparticu-
lates [22,23], among others. The objective of this review is
to identify the specific strategic directions and challenges
that nanovectors can solve, and, within this context,
provide a brief overview of leading nanovector technol-
ogies for drug delivery. To simplify the presentation, the
main focus is placed on cancer therapeutics, but confi-
dence in the applicability of progresses to other branches
of medicine is implied.
Defining the crucial pathways for nanovectors
The fundamental breakthrough opportunities for nano-
vector delivery are summarized in three, closely inter-
related main aspects: first, the recognition of target cells
and tissues; second, the ability to reach the diseased sites
where the target cells and tissues are located; and third,
the ability to deliver multiple therapeutic agents. The
first two aspects comprise the notion of achieving pre-
ferred, substantially higher concentration of therapeutic
action at lesion sites, a phenomenon that will be called
‘localization’ in this article, as opposed to the term ‘tar-
geting’ that is often used to identify drugs that provide
specific action against a target biological pathway. Each of
the three fundamental aspects further articulates into
several statements of challenge, and is accordingly pre-
sented in what follows.
Recognizing intended cells and tissues
The most widely investigated modalities for the recogni-
tion of the cells and tissues against which the therapy is
directed involve the attachment of therapeutic agents (in
nanoparticle format or not) to biological recognition
Current Opinion in Chemical Biology 2005, 9:343–346
344 Next-generation therapeutics
moieties, such as antibodies, aptamers, substrates and
ligands. Although successful in some instances, this
approach presents limitations on at least three fronts:
first, the specificity of the recognition agent; second,
the relative localizing overexpression of the conjugate
‘target’ molecules expressed, for instance, as cell-surface
antigens of microenvironmental markers such as integrins
on neovascular endothelium; and third, the fact that the
biological recognition agents are themselves mostly macro-
molecules, and the object of adverse action by the natural
defenses of the body, as discussed in the next section. Even
therapeutic antibodies by themselves reach their intended
targets only in the ratios of 1 in 10 000 to 1 in 100 000. While
improvement in biological targeting is certainly an impor-
tant direction to pursue, and is being actively investigated
by the nanotechnology community [8,13,24,25–27], the
use of nanovector delivery affords substantial advantages
for what pertains to a broader consideration of target
selectivity. These are obtained from the decoupling of
the therapeutic action per se from the localization require-
ments — the latter are left to the nanovector, as opposed to
the drug they carry.
The key strategic direction is thus the use of multimodal
localization strategies, of which biological affinity-based is
but one, on the basis that localization probabilities are
essentially additive: as long as they are non-interfering,
each contributes favorably to the achievement of preferred
spatial concentrations. Some promising non-biomolecular
targeting mechanisms are briefly discussed next.
Localization by size and shape
Years of liposome research and clinical use have demon-
strated that tailoring nanovector size to match the fenes-
trations of the cancer neovasculature yields preferential
concentration at tumor sites — a phenomenon termed
enhanced penetration and retention (EPR). Yet, the
spherical shape of liposomes, and sizes of about 50–
100 nm tend to keep them in the middle of blood flow
[8,9,10–13,14,15,16–19,20,21–23,24,25–28,29] as
opposed to the vicinity of the endothelial wall, and there-
fore diminish the likelihood of convection through the
fenestrations, and molecular targeting to the neovascular
endothelium. Similar considerations apply to the near
totality of nanovectors presented in the literature, since
they are all spherical or nearly spherical. Recent advances
in nanofabrication technology, however, may open the
way towards the development of more suitable geome-
tries for injectable, silicon- and polymer-based delivery
systems (Decuzzi P and Ferrari M, unpublished data).
Localization by physical properties
The recognition that the surface charge of nanovectors
influences tumor uptake is not very recent [30,31] but has
been essentially forgotten by the community. Contem-
porary mathematical models have advanced the under-
standing of the relationships between surface charge and
Current Opinion in Chemical Biology 2005, 9:343–346
stromal accumulation [29], and may induce the devel-
opment of novel classes of nanovectors.
Localization by remote or environmental activation
Regardless of the details of the distribution of injected
nanovectors in the body, exquisite localization of the
effect may be attained if the cytotoxic action is released
only at the intended target sites, by irradiation with an
exogenous, and locally focused source of energy. Remote
activation approaches that have been demonstrated in the
literature include the triggering of gold nanoshells by
near-infrared radiation [9], leading to localized thermal
ablation of tumor xenografts in animal models [10–12].
Enhanced photodynamic therapy by targeted silica nano-
particles [21,32], neutron-capture therapy with gadoli-
nium nanoparticles [23], the ultrasound-based delivery
from lipid-encapsulated microbubbles [33] and the mag-
netic field activation of cytolysis [34] have been reported.
Localized release may also be activated by environmental
conditions, such as metabolic markers [15], and the
acidity levels that accompany inflammatory states, infec-
tions, and neoplastic processes [35]. Therapeutic target-
ing may be attained by two-step activation methodologies
such as polymer-directed enzyme prodrug therapy
(PDEPT) and polymer-enzyme liposome therapy
(PELT) [2].
Reaching the target lesions
No level of targeting sophistication will produce substan-
tial benefits in therapeutic index, unless the agents of the
therapeutic actions can reach the intended lesions sites.
Intrinsic to the body defense systems are several extre-
mely effective obstacles (collectively termed ‘biobar-
riers’) that largely prevent injected chemicals,
biomolecules, nanoparticles and any other foreign agents
of therapeutic action from reaching their intended desti-
nations. Biobarriers are sequential in nature, and there-
fore the probability of reaching the therapeutic objective
is the product of the individual probabilities of over-
coming each one of them. A corollary is that any efficient
delivery method must be provided with tools that allow it
to overcome all of these barriers, because opening all of
the doors but one along a single path will obviously not
suffice. Requiring a therapeutic agent to be provided with
a sufficient collection of weaponry to conquer all barriers
and still be small enough for safe vascular injection is a
challenge that only nanotechnology can meet, by defini-
tion. Multiple barrier-avoidance methodologies are under
investigation. Some of the leading developments are
summarized next, categorized by bio-barrier typology.
Endothelial and epithelial barriers
The blood–brain barrier (BBB) poses a formidable obsta-
cle to penetration by therapeutic agents. Iron-oxide nano-
particles, however, by way of their physical properties,
have demonstrated considerable efficacy in reaching
brain tumors [36–38], and serve locally as signal enhancers
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for the pre-, post- and intra-operative mapping of cancer
lesions by magnetic resonance and optical imaging means
[39–41]. These particles are therefore promising candi-
dates as vectors for combined therapeutics and diagnostics.
An example of nanotechnologies for overcoming epithelial
barriers involves the co-localized delivery of a therapeutic
biomolecule with a penetration enhancer, such as a zonula
occludens toxin (ZOT). This acts to open intracellular
tight junctions in a short-term, reversible and localized
fashion, thus affording transport of the therapeutic bio-
molecule into the vascular compartment, without concur-
rent increased risks of opportunistic infection [42].
Sequestration by the reticulo-endothelial system
The circulatory clearance half-time of liposomes [26,43]
was increased from minutes to hours or days by the
attachment of polyethylene glycol (PEG) to their surfaces.
PEG provides a shielding ‘stealth’ effect, delaying recog-
nition and sequestration by the resident macrophages of
the reticulo-endothelial system. Unfortunately, this
‘immunostealthing’ function is concurrent with the anni-
hilation of biomolecular targeting capabilities, because the
PEG molecules hide localizing antibodies conjugated on
the liposomal surfaces. PEG may be expected to increase
the circulatory time of other nanovectors as well, but new
strategies are needed to provide an increase in clearance
time without detriment to lesion localization.
Adverse osmotic pressure
As cancer lesions grow, they develop an increased internal
hydrostatic pressure that counters convective extravasa-
tion from the vascular compartment into the tumor. This
leaves diffusion as the only mechanism of transport of
therapeutic agents into the tumor — a very unlikely route
for large molecules or nanovectors. This is a very central
problem, which unfortunately has not been convincingly
addressed in the literature to date. A speculative approach
based on the use of molecular motors as active transpor-
ters was recently presented [44].
Delivering multiple agents of therapy
Contemporary cancer therapeutics generally involve the
simultaneous use of multiple drugs, based on the need to
intervene on a spectrum of cancer-overactive mechanisms
possibly acting in a parallel or redundant fashion. The
probability of two different drugs reaching the same site is
obtained by multiplying the individual probabilities, at a
massive increment of potential side effects. In an over-
simplified computational example, two drugs with a prob-
ability of 1 in 1000 of reaching the intended target only
combine to act with a probability of 1 in 1 000 000. At the
same time, essentially twice the amount of active prin-
ciples reaches unintended sites than would with either
one of the agents alone, and thus causes much greater
undesired collateral effects. This argues in favor of co-
localizing multi-drug therapies by incorporation in nano-
vector carriers — an approach that has not yet been
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Nanovector therapeutics Ferrari 345
substantially articulated in the literature. However, the
benefits of combined nanovectored treatment by liposo-
mal gene delivery and radiation or chemotherapy, has
been convincingly illustrated [26].
Conclusions
Based on clinical evidence on liposomes to date, and a
great wealth of recent technological advances in nanos-
cale manufacturing and synthesis, nanovectors yield sub-
stantial promise for the development of highly beneficial
and innovative methods for the treatment of cancer and
other pathologies. The opportunities for nanovector ther-
apeutics depend on the successful addressing of the three
critical challenges: first, the development of multimodal
localization capabilities, that exploit multiple, concurrent
mechanisms to obtain preferential concentrations of the
therapeutic action at the intended lesion sites; second, the
integration in nanovectors of multiple methods for the
sequential overcoming of the natural defenses that create
barriers against the reaching of the intended lesions,
regardless of their intrinsic targeting abilities; and third,
the ability of comprising multiple therapeutic agents
within nanovectors, to attain their synergistic co-delivery
at lesion sites.
Significant progress has been reported in the literature
toward addressing the first two of these challenges, but
attention to the development of a system that incorpo-
rates all of the desired functions has been minimal. While
the present review has focused explicitly on cancer and
the delivery of therapeutics agents, the considerations
presented may be extended beyond oncology, to embrace
gene delivery, and preventative medicine.
Acknowledgements
The author is grateful to the National Cancer Institute, the State of Ohio
BRTTF, and The Ohio State University for their support of this work.
References and recommended reading
Papers of particular interest, published within the annual period of
review, have been highlighted as:
 of special interest
 of outstanding interest
1. Becker ML, Wooley KL: Peptide-polymer bioconjugates: hybrid
block copolymers generated via living radical polymerizations
from resin-supported peptides. Chem Commun (Camb)
2003:180-181.
2. Duncan R: The dawning era of polymer therapeutics. Nat Rev
 Drug Discov 2003, 2:347-360.
A comprehensive review of polymer-based delivery system, with insights
into future potential breakthrough approaches.
3. La Van DA, McGuire T, Langer R: Small-scale systems for in-vivo
drug delivery. Nat Biotechnol 21, 10:184-1191, 2003.
4. Nakanishi T, Fukushima S, Okamoto K, Suzuki M, Matsumura Y,
Yokoyama M, Okano T, Sakurai Y, Kataoka K: Development of
the polymer micelle carrier system for doxorubicin. J Control
Release 2001, 74:295-302.
5. Soppimath KS, Rudzinski WE: Biodegradable polymeric
nanoparticles as drug delivery devices. J Control Release 2001,
70:1-20.
Current Opinion in Chemical Biology 2005, 9:343–346
346 Next-generation therapeutics
6. Cloninger MJ: Biological applications of dendrimers. Curr Opin
Chem Biol 2002, 6:742-748.
7. Gilles ER, Frechet JMJ: Designing macromolecules for
therapeutic applications: Polyester dendrimer-polyethylene
oxide ‘bow-tie’ hybrids with tunable molecular weights and
architecture. J Am Chem Soc 2002, 124:14137-14146.
8. Quintana A, Baker JN Jr: Design and function of a
dendrimer-based therapeutic nanodevice targeted to
tumor cells through the folate receptor. Pharm Res 2002,
19:1310-1316.
9. Charnay C, Halas NJ: Reduced symmetry metallodielectric
 nanoparticles: chemical synthesis and plasmonic properties.
J Phys Chem B 2003, 107:7327-7333.
Insight into the fabrication of gold nanoshells may yield inspiration for the
mathematical modeling of their plasmonic response, towards tunability of
the system.
10. Hirsch LR, Halas NJ, West JL: Nanoshell-mediated near-
infrared thermal therapy of tumors under magnetic resonance
guidance. Proc Natl Acad Sci USA 2003, 100:13549-13554.
11. Loo C, Halas NJ, West J, Drezek R: Nanoshell-enabled
photonics-based imaging and therapy of cancer. Technol
Cancer Res Treat 2004, 3:33-40.
12. O’Neal DP, Halas NJ, West JL: Photo-thermal tumor ablation in
mice using near infrared-absorbing nanoparticles. Cancer Lett
2004, 209:171-176.
13. Akerman ME, Chan WCW, Laakkonen P, Bhatia SN, Ruoslahti E:
Nanocrystal targeting in vivo. Proc Natl Acad Sci USA 2002,
99:12617-12621.
14. Derfus AM, Chan WCW, Bhatia SN: Probing the cytotoxicity of
 semiconductor quantum dots. Nano Lett 2004, 4:11-18.
Nanoparticle biocompatibility is a fundamental concern. The methods
displayed in this paper may guide researchers working with different
nanoparticle formulations, as well.
15. Ishii D, Aida T: Chaperonin-mediated stabilization and
 ATP-triggered release of semiconductor nanoparticles.
Nature 2003, 423:628-632.
Adapting nanovector design to attain preferential, locally triggered acti-
vation offers potential advantages in therapeutic index.
16. Raja KS, Manchester M, Finn MG: Hybrid virus-polymer
materials. 1. Synthesis and properties of PEG-decorated
cowpea mosaic virus. Biomacromolecules 2003, 4:472-476.
17. He XX, Li J: Bioconjugated nanoparticles for DNA protection
from cleavage. J Am Chem Soc 2003, 125:7168-7169.
18. Vijayanathan V, Thomas TJ: DNA nanoparticles and
development of DNA delivery vehicles for gene therapy.
Biochemistry 2002, 41:14085-14094.
19. Cohen MH, Melnik K, Boiarski A, Ferrari M, Martin FJ:
Microfabrication of silicon-based nanoporous particulates for
medical applications. Biomedical Microdevices 2003, 5:253-259.
20. Ferrari M: Therapeutic Microdevices and Methods of Making and
 Using Same. US Patent No. 6,107,102, August 22, 2000.
Multiple concepts and designs for injected delivery systems are presented.
21. Yan F, Kopelman R: The embedding of meta-
tetra(hydroxyphenyl)-chlorin into silica nanoparticle platforms
for photodynamic therapy and their singlet oxygen production
and pH-dependent optical properties. Photoch Photobiol 2003,
78:587-591.
22. Yan F, Kopelman R, Reddy R: Synthesis and characterization of
silica-embedded iron oxide nanoparticles for magnetic
resonance imaging. J Nanosci Nanotechnol 2004, 4:72-76.
23. Oyewumi MO, Mumper RJ: Engineering tumor-targeted
gadolinium hexanedione nanoparticles for potential
application in neutron capture therapy. Bioconjug Chem 2002,
13:1328-1335.
24. Li L, Wartchow CA, Danthi SN, Shen Z, Dechene N, Pease J, Choi
 HS, Doede T, Chu P, Ning S: A novel antiangiogenesis therapy
using an integrin antagonist or anti-Flk-1 antibody coated
90Y-labeled nanoparticles. Int J Radiat Oncol Biol Phys 2004,
58:1215-1227.
Current Opinion in Chemical Biology 2005, 9:343–346
Nanoparticles are generally excluded from extravasation through healthy
vasculature, because of their ‘large’ size. This actually presents an
opportunity: improved conditions for the localization on neoplastic
endothelium, and the delivery of antiangiogenesis therapy thereon (see
also [20,27]).
25. Pan D, Wooley KL: Folic acid-conjugated nanostructured
materials designed for cancer cell targeting. Chem Commun
(Camb) 2003:2400-2401.
26. Pirollo KF, Xu L, Chang EH: Immunoliposomes: a targeted
delivery tool for cancer treatment. In Vector Targeting for
Therapeutic Gene Delivery. Curiel D(Ed) Wiley-Liss; 2002.
27. Winter PM, Wickline SA, Lanza GM: Molecular imaging of
angiogenesis in nascent Vx-2 rabbit tumors using a novel
alpha(nu)beta3-targeted nanoparticle and 1.5 Tesla magnetic
resonance imaging. Cancer Res 2003, 63:5838-5843.
28. Decuzzi PS, Decuzzi LM, Ferrari M: Adhesion of micro-
fabricated particles on vascular endothelium: a parametric
analysis. Annals Biomed Eng 2004, 32:793-802.
29. Decuzzi P, Lee S, Bhushan B, Ferrari M: Non-specific interaction
 of nanoparticles as drug delivery and nanoharvesting agents
within the vasculature. Annals Biomed Eng 2005, in press.
Mathematics will be increasingly important for bringing nanotechnology
to full fruition in medicine.
30. Juliano RL, Stamp D: Effect of particle size and charge on the
clearance rates of liposomes and liposome-encapsulated
drugs. Biochem Biophys Res Commun 1975, 63:651-658.
31. Dellian M, Yuan F, Trubetskoy VS, Torchilin VP, Jain RK: Vascular
permeability in a human tumor xenograft: molecular charge
dependence. Br J Cancer 2000, 82:1513-1518.
32. Roy I, Bergey EJ, Prasad PN: Ceramic-based nanoparticles
entrapping water-insoluble photosensitizing anticancer
drugs: a novel drug-carrier system for photodynamic therapy.
J Am Chem Soc 2003, 125:7860-7865.
33. May DJ, Allen JS, Ferrara KW: Dynamics and fragmentation of
thick-shelled microbubbles. IEEE Transactions on Ultrasonics
Ferroelectrics & Frequency Control 2002, 49:1400-1410.
34. Bergey EJ, Prasad PN: DC magnetic field induced
magnetocytolysis of cancer cells targeted by LH-RH magnetic
nanoparticles in vitro. Biomedical Microdevices 2002, 4:293-299.
35. Potineni A, Langer R, Amiji MM: Poly(ethylene oxide)-modified
poly(beta-amino ester) nanoparticles as a pH-sensitive
biodegradable system for paclitaxel delivery. J Control Release
2003, 86:223-234.
36. Koziara JM, Allen DD, Mumper RJ: In situ blood-brain barrier
transport of nanoparticles. Pharm Res 2003, 20:1772-1778.
37. Lockman PR, Allen DD: Nanoparticle technology for drug
deliver across the blood-brain barrier. Drug Dev Ind Pharm
2002, 28:1-13.
38. Lockman PR, Allen DD: Brain uptake of thiamine-coated
nanoparticles. J Control Release 2003, 93:271-282.
39. Kircher MF, Mahmood U, King RS, Weissleder R, Josephson L: A
multimodal nanoparticle for preoperative magnetic resonance
imaging and intraoperative optical brain tumor delineation.
Cancer Res 2003, 63:8122-8125.
40. Neuwelt EA, Varallyay P, Bago AG, Muldoon LL, Nesbit G, Nixon R:
Imaging of iron oxide nanoparticles with MR and light
microscopy in patients with malignant brain tumors.
Neuropathol Appl Neurobiol 2004, 30:456-471.
41. Steiniger SC, Gelperina SE: Chemotherapy of glioblastoma in
rats using doxorubicin-loaded nanoparticles. Int J Cancer
2004, 109:759-767.
42. Ferrari M, Martin F, Grove C: Particles for Oral Delivery of Peptides
and Proteins. US Patent No. 6,355,270, March 12, 2002.
43. Park JW: Liposome-based drug delivery in breast cancer
treatment. Breast Cancer Res 2002, 4:95-99.
44. Ferrari M: Cancer nanotechnology: opportunities and
challenges. Nat Rev Cancer 2005, 5:161-171.
www.sciencedirect.com