Advanced Drug Delivery Reviews 64 (2012) 270–279

Contents lists available at SciVerse ScienceDirect

Advanced Drug Delivery Reviews

journal homepage: www.elsevier.com/locate/addr

Advanced drug delivery devices via self-assembly of amphiphilic block copolymers☆

Annette Rösler, Guido W.M. Vandermeulen, Harm-Anton Klok ⁎
Max-Planck-Institute for Polymer Research, Ackermannweg 10, D-55128 Mainz, Germany

a r t i c l e i n f o a b s t r a c t

Available online 13 September 2012 Amphiphilic block copolymers are well established as building blocks for the preparation of micellar drug carriers.
Over the past decade, the effectiveness of such self-assembled drug delivery devices has been demonstrated
Keywords: numerous times. This review will discuss two approaches that can be used to further improve the effectiveness
Block copolymer micelle of amphiphilic block copolymer-based drug delivery systems. The first approach involves the chemical modifica-
Nanocapsule tion of the block copolymer building blocks. Several examples will be discussed of amphiphilic block copolymers
Crosslinking
modified with crosslinkable groups in order to increase the stability of the micellar drug carriers, or of block
Targeted drug delivery
Auxiliary agent
copolymers containing specific ligands that could ultimately allow targeted drug delivery. The second approach
Channel Protein to improve the performance of micellar drug carriers is the addition of auxiliary agents. To illustrate this approach,
Metal particle the feasibility of channel proteins and metal (nano)particles to improve temporal control over the drug release
process is discussed.
© 2012 Published by Elsevier B.V.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 270
0
2. Functional block copolymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 271
0
2.1. Core crosslinkable micelles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 272
0
2.2. Shell crosslinkable micelles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275
0
2.3. Surface functionalized micelles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 276
0
3. Auxiliary agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 276
0
3.1. Channel proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 277
0
3.2. Metal (nano)particles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278
0
4. Conclusions and outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278
0
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278
0
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278
0

1. Introduction transported at concentrations that can exceed their intrinsic water-

The continuous development of new drug delivery systems is driven
by the need to maximize therapeutic activity while minimizing negative
side effects. One class of drug delivery vehicle that has received
widespread attention over the past decades is micelles formed by
self-assembly of amphiphilic block copolymers in aqueous solution
[1–3]. Block copolymer micelles are of interest for drug delivery applica-
tions for a number of reasons. First of all, hydrophobic drugs can
be physically entrapped in the core of block copolymer micelles and
☆ PII of original article: S0169-409X(01)00222-8. The article was originally published
in Advanced Drug Delivery Reviews 53 (2001) 95–108.
⁎ Corresponding author. Tel.: +49 6131 379 306; fax: +49 6131 379 100.
E-mail address: hak@mpip-mainz.mpg.de (H.-A. Klok).
solubility. Secondly, the hydrophilic blocks, which are often composed
of poly(ethylene oxide) (PEO), can form hydrogen bonds with the
aqueous surroundings and form a tight shell around the micellar core.
Diblock copolymer micelles with a PEO corona resist protein ad-
sorption and cellular adhesion. As a result, the contents of the hydropho-
bic core are effectively protected against hydrolysis and enzymatic
degradation. In addition, the PEO corona prevents recognition by the
reticuloendothelial system and therefore preliminary elimination of the
micelles from the bloodstream. Thus, these so-called ‘stealth’ properties
of the PEO corona result in increased blood circulation times and
allow drugs to be administered over prolonged periods of time. A final
feature that makes amphiphilic block copolymers attractive for drug
delivery applications is the fact that their chemical composition, total
molecular weight, and block length ratios can be easily changed, which
allows control of the size and morphology of the micelles [4,5].

0169-409X/$ – see front matter © 2012 Published by Elsevier B.V.

http://dx.doi.org/10.1016/j.addr.2012.09.026
 A. Rösler et al. / Advanced Drug Delivery Reviews 64 (2012) 270–279
Two important issues when discussing the effectiveness of a drug
therapy are the extent to which temporal and distribution control can
be achieved [6]. Temporal control refers to the ability to adjust the
period of time over which drug release is supposed to take place or
to the possibility to trigger the release process at a specific time
during treatment. The aim of distribution control is to precisely direct
the drug delivery system to the desired site of activity. A very elegant
approach to improve temporal control involves the use of block
copolymers where one of the segments possesses a lower-critical
solution temperature (LCST). Below their LCST, polymers such as
poly(N-isopropylacrylamide) (PNIPAAm) are water-soluble, while
raising the temperature of an aqueous solution above the LCST results
in phase separation. Based on this concept, several PNIPAAm containing
block copolymers have been reported, which were used to prepare
thermosensitive micellar drug carriers [7–9]. Changing the temperature
of the environment slightly above or below the LCST can result in desta-
bilization of the micelle and trigger a burst-like release of the encapsu-
lated drugs. Although this approach is clinically feasible, it has the
disadvantage that the block copolymer micelles cannot be directly
addressed and the body temperature of the patient needs to be locally
increased or decreased. Distribution control, especially with respect to
the treatment of solid tumors, has been achieved by taking advantage
of the enhanced vascular permeability of tumor tissue compared to
healthy tissue [10–12]. As a result, high molecular weight substances
can be preferentially accumulated at these sites. This is an example of
passive drug targeting, which is referred to as the enhanced permeabil-
ity and retention effect (EPR).
In spite of the advances discussed above and the successful imple-
mentation of block copolymer-based drug delivery devices there is still
a need to further improve the performance of these systems both with
respect to temporal as well as distribution control. These issues have
gained increasing importance with the recent developments in the
Fig. 1. Schematic representation of the three different classes of functional amphiphilic block copolymers discussed. Depending on the type of functionalization, either shell
crosslinked micelles, core crosslinked micelles or surface functionalized micelles can be obtained.
271
design of peptide drugs and the emergence of gene therapy. The clinical
success of these very potent pharmaceutical agents will strongly depend
on the possibility to precisely direct the drug to the desired site of activity
in the body and to accurately control the rate at which the drug is
released and the time period over which the drug is administered.
During the past years, an increasing number of publications has
appeared that describe the development of advanced drug delivery
systems with the aim of further improving temporal and/or distribu-
tion control of the release process. This review highlights some of the
recent developments. Due to the large number of papers that has
been published and the limited space available for this review it is
virtually impossible to give a complete overview of the field. Rather,
we decided to select two different approaches for the development
of advanced block copolymer-based drug delivery devices and illus-
trate these concepts with examples from the recent literature. In the
remaining part of this review we will successively discuss the use of
functional block copolymers and the use of auxiliary agents.
Functionalization of block copolymers with crosslinkable groups
can increase the stability of the corresponding micelles and improve
temporal control. Substitution of block copolymer micelles with specific
ligands is a very promising strategy to target a broader range of sites of
activity with a much higher selectivity in comparison with the EPR effect
discussed above. The use of appropriate auxiliary agents can allow direct
addressing of the block copolymer micelles, without the need for hyper-
thermia or hypothermia.
2. Functional block copolymers
Functional block copolymers designed to improve the perfor-
mance of self-assembled drug delivery systems can be divided into
three major classes (Fig. 1). The first class of block copolymers con-
tains one or more functional groups on the hydrophobic block,
272 A. Rösler et al. / Advanced Drug Delivery Reviews 64 (2012) 270–279
which allow crosslinking of the core of the micelle. In the second class
of block copolymers these substituents are located on the hydrophilic
part of the molecule and enable crosslinking of the micellar
corona. The third class of block copolymers is surface functionalized
with ligands that allow the drug delivery device to be directed to
the specific site of interest.
Chemical fixation of micelles by crosslinking of either the core or
the corona is of interest for a number of reasons. Crosslinking gener-
ally increases the stability of the micelles. Crosslinked micelles are
stable at concentrations below the critical micelle concentration
(CMC) of the block copolymer, can be isolated and redissolved as
stable nanoparticles and are less likely to collapse, for example in
the blood stream. As a result, crosslinking can lead to increased circu-
lation times, allowing drugs to be administered over longer periods of
time. Crosslinking thus provides a means to improve temporal con-
trol. In the case of shell-crosslinked micelles, however, the effect of
crosslinking is two-fold. First of all, as discussed above, the stability
of the micelle increases. In addition, crosslinking will also affect the
permeability of the corona, and therefore might provide a means to
fine-tune the rate at which drugs can be released from the micellar
core. The aim of functionalizing block copolymers with ligands that
help target a specific site of interest is obvious and clearly contributes
to improving distribution control of the drug delivery process.
Fig. 2. The preparation of well-defined layers of surface-bound PEG–PLA micelles via alternate deposition of aldehyde functionalized PEG–PLA micelles and polyallylamine (adapted
from Ref. [17]).
Below the three different approaches towards functional block
copolymers illustrated in Fig. 1 will be discussed with some recent
examples. At this point it is worthwhile mentioning that block
copolymers can also contain features that are typical for more than
one of the classes introduced above. Although this makes it difficult
to classify these materials, this is of course a very effective strategy
to tailor the properties of block copolymer-based drug delivery
systems.
2.1. Core crosslinkable micelles
There are various possibilities to functionalize amphiphilic block
copolymers in such a way that they will allow crosslinking of the
core of the micelle. Probably the most suitable method for drug
delivery purposes is to modify the hydrophobic chain end of the
block copolymer with a polymerizable group. In this case, the micellar
stability can be increased by crosslinking of the functional groups
without affecting the drug loading capacity, i.e. the hydrophobic
space available to encapsulate drugs. Alternative strategies to achieve
core crosslinking include the introduction of polymerizable groups
along the hydrophobic segment of the block copolymer or the encap-
sulation and polymerization of low molecular weight monomer in the
hydrophobic core of the micelle. The last two methods, however, have
 A. Rösler et al. / Advanced Drug Delivery Reviews 64 (2012) 270–279
Fig. 3. Schematic representation of the formation of reversibly crosslinked micelles from thiol-modified PEG–PLL block copolymers (adapted from Ref. [20]).

273
274 A. Rösler et al. / Advanced Drug Delivery Reviews 64 (2012) 270–279
Fig. 4. The formation of shell-crosslinked block copolymer micelles via the self-assembly of amphiphilic block copolymers. Degradation and removal of the core of these micelles
results in a polymeric nanocontainer or nanocage (adapted from Ref. [23]).
the disadvantage that the crosslinking reaction will reduce the free
volume of the hydrophobic core, resulting in a decrease in the drug
loading capacity.
Block copolymers having a polymerizable group at their hydro-
phobic chain end have been extensively studied by Kataoka and
coworkers [13–17]. These authors have reported a number of poly(D,
L-lactide)-b-poly(ethylene glycol) (PLA–PEG) copolymers containing a
methacryloyl group at the PLA chain end. After micellization the
methacryloyl groups could be polymerized either thermally by adding
a radical initiator or under the influence of UV light in the presence of
a photo initiator. The size of the resulting core crosslinked micelles
was constant up to 60 °C. These core crosslinked micelles did not col-
lapse upon dissolution in non-selective organic solvents and also
resisted treatment with sodium dodecyl sulfate.
In a number of cases, heterobifunctional PLA–PEG block copolymers
have been reported which carry a crosslinkable group at the PLA chain
end and an aldehyde group at the PEG terminus. Coating of these
micelles on an amine functionalized surface in the presence of NaCNBH3
resulted in fixation of the PEG chain end on the surface [15]. In
the case of crosslinked PLA–PEG micelles, the core-shell structure
was maintained even after chemical fixation to the substrate, while
the non-polymerized PLA–PEG micelles disrupted when they were
attached to the substrate. Such surface tethered micelles might also be
used to entrap and release drugs. As is shown in Fig. 2, the surface
concentration and thus the drug loading capacity of such surface-
bound micelles can be nicely controlled by alternate deposition of
PLA–PEG diblock copolymer micelles and polyallylamine [16,17]. It
was found that such multilayer structures prevent adsorption of
proteins and can release hydrophobic reagents in a controlled manner.
Several authors have explored amphiphilic block copolymers
containing crosslinkable groups laterally distributed along the hydro-
phobic block. Bates et al., for example, described the chemical fixation
of worm-like poly(ethylene oxide)-b-poly(butadiene) (PEO–PB)
micelles via crosslinking of the PB core [18]. The use of a redox ini-
tiating system of potassium peroxodisulfate (K2S2O8) and sodium
disulfite (Na2S2O5)-ferrosulfate (FeSO4 · 7H2O) resulted in quantitative
crosslinking without affecting the morphology of the micelles. Follow-
ing the same strategy of crosslinking lateral substituents at the hydropho-
bic block, Henselwood and Liu prepared water-soluble nanospheres by
UV irradiation of poly(2-cinnamoylethyl methacrylate)-b-poly(acrylic
acid) (PCEMA–PAA) block copolymers [19]. Since these nanospheres
were intended to be used for the uptake of organics from aqueous solu-
tion, the crosslinking density of the PCEMA block was deliberately kept
low.
A very elegant example of the use of laterally functionalized
amphiphilic block copolymers, more directly related to drug delivery
applications, was described by the group of Kataoka [20,21]. By the
introduction of thiol groups on a small fraction (10–20%) of the lysine
 A. Rösler et al. / Advanced Drug Delivery Reviews 64 (2012) 270–279
Fig. 5. Chemical structures of three differently end-functionalized diblock copolymers (top) and the elution profiles that were recorded after passing micellar solutions of these
block copolymers through an RCA-1 functionalized affinity column (bottom) (adapted from Ref. [35]).
repeat units of poly(ethylene glycol)-b-poly(L-lysine) (PEG–PLL) diblock
copolymers, reversibly crosslinkable micelles could be prepared. The
concept pursued by Kataoka et al. is illustrated in Fig. 3.
Upon the addition of a negatively charged polyelectrolyte, such
as an oligonucleotide (ODN) or poly(aspartic acid), the thiolated
PEG–PLL diblock copolymers form so-called polyion complex
micelles (PICs). Under ambient conditions, oxidation of the thiol
groups results in the formation of disulfide bonds and crosslinking
of the micellar core. The unique feature of disulfide crosslinks is
that they can be broken under the influence of an appropriate
reducing agent. Two types of reducing agents were investigated:
(i) dithiothreitol (DTT) and (ii) glutathione (GSH). When applied
under the appropriate conditions, both reagents could induce disso-
ciation of the micelles, resulting in release of the encapsulated poly-
electrolyte. Of the two investigated reducing agents GSH is probably
the most interesting as it is the most abundant reducing agent in
cells. The intracellular GSH concentration is 300 times higher than
the concentration in the blood. Kataoka et al. demonstrated that
the PICs were stable to low concentrations of GSH, whereas increas-
ing the GSH concentration to the millimolar range, as present in
cells, resulted in dissociation of the micelles and release of the
entrapped ODN. These experiments clearly indicate the potential
of these block copolymer micelles as carriers for DNA in gene
therapy.
2.2. Shell crosslinkable micelles
One of the first examples of the concept of chemically crosslinking
the corona of amphiphilic diblock copolymer micelles was reported by
Wooley and coworkers in 1996 [22]. Since then, this strategy has been
elaborated in detail and has been picked up by a number of other groups
as well [23–33]. The synthetic route towards shell-crosslinked block co-
polymer micelles (or ‘shell crosslinked knedels’, as they were termed by
Wooley) is outlined schematically in Fig. 4.
The chemistry involved in the preparation of shell-crosslinked
block copolymer micelles is basically limited to the preparation of
an appropriate amphiphilic diblock copolymer. As a result, advantage
can be taken from the versatility of block copolymer chemistry to
tune the structure and chemical composition of the crosslinked
275
micelles. For the synthesis of the hydrophobic core-forming block, a
broad range of polymers has been used, including, for example, sty-
rene, isoprene, butadiene, ε-caprolactone and methylmethacrylate.
Monomers that have been employed for the preparation of the hydro-
philic, water-soluble corona include, amongst others, 4-vinylpyridine,
methacrylic acid and 2-dimethylaminoethyl methacrylate. The coro-
na of the micelles is chemically fixed in aqueous solution after
self-assembly of the amphiphilic diblock copolymer. Crosslinking
has been accomplished with covalent bonds, ionic bonds and a com-
bination of the two. Poly(acrylic acid)-based systems are typically
crosslinked by the addition of a diamine under conditions also used
in peptide chemistry [23–26]. 2-(Dimethylaminoethyl)methacrylate
containing shells have been crosslinked via quaternization with
1,2-bis(2-iodoethoxy)ethane [27,28]. Crosslinked poly(isoprene)- or
poly(styrene)-b-poly(2-cinnamoylethyl methacrylate) shells have
been prepared by UV irradiation [29–31]. Micellar coronas composed
of 4-vinylpyridine have been crosslinked by quaternization with
p-chloromethylstyrene, followed by photoinduced radical polymeri-
zation of the styrene double bonds [22,32].
As shown in Fig. 4, the strategy discussed above is not limited
to shell-crosslinked micelles, but can, by removal of the core, be ex-
tended to the preparation of nanosized hollow particles [23–26,30].
Such nanocontainers might be of particular interest for drug delivery
applications due to their potentially higher loading capacity as com-
pared with the micellar precursors and due to the fact that they
might provide an interior suitable for the transport and controlled
release of hydrophilic guests that cannot be encapsulated in the
hydrophobic interior of most water-soluble amphiphilic diblock
copolymer micelles. Removal of the core of shell-crosslinked micelles
has been accomplished in a number of ways, including ozonolysis
of poly(isoprene) [24,30], UV photolysis of poly(silanes) [25], acid
catalyzed hydrolysis of poly(ε-caprolactone) [26] and thermolysis
of a C–ON bond connecting the poly(styrene) and poly(acrylic acid)
segments of a block copolymer obtained via a combination of
nitroxide-mediated and atom transfer radical polymerization [23].
Although shell-crosslinked block copolymer micelles and their
corresponding nanocages have only been developed recently, and
detailed studies with respect to their feasibility for drug delivery
applications are lacking at this moment, it is undeniable that they
276 A. Rösler et al. / Advanced Drug Delivery Reviews 64 (2012) 270–279

Fig. 6. Schematic representation of a PMOXA–PDMS–PMOXA vesicle containing the enzyme β-lactamase. (a) In the absence of a channel protein, no transport of small molecules
into and out of the vesicle is possible. (b) After incorporation of the channel protein (OmpF), transport of small molecules into and out of the vesicle becomes possible. This is
evidenced by the detection of ampicillinoic acid, which is the product of the reaction of ampicillin with the enzyme. The OmpF channels can be closed by the addition of a
polyelectrolyte. This process can be reversed by the addition of a low molecular weight electrolyte or by simple dilution (b↔c) (adapted from Ref. [37]).

possess great potential for the development of advanced drug deliv-
ery systems. These expectations are inspired by first experiments
that suggest that control of release rates as well as active drug
targeting may become possible by appropriately preparing and/or
functionalizing the nanocages [23,33].
2.3. Surface functionalized micelles
The core and shell crosslinking strategies discussed above can lead
to an increased stability of the micelle and improve temporal control
of the drug release process. An equally important aspect, however, is
the ability to direct the drug loaded micelle to the site of interest
(distribution control). Two major mechanisms can be distinguished
for addressing the desired sites for drug release: (i) passive targeting
and (ii) active targeting. An example of passive drug targeting was
mentioned in the Introduction and involves the preferential accumu-
lation of chemotherapeutic agents in solid tumors as a result of the
enhanced vascular permeability of tumor tissues compared with
healthy tissue. A strategy that could allow active targeting involves
the modification of the chain end of the hydrophilic part of the
block copolymer with ligands that are selectively recognized by
receptors on the surface of the cells of interest. Since ligand–receptor
interactions can be highly selective this could allow a more precise
targeting of the site of interest. In addition, active targeting is not
restricted to, for example, solid tumors, but is applicable to a much
broader range of targets.
Some examples of amphiphilic diblock copolymers functionalized
at their hydrophilic chain end with a ligand that can mediate recogni-
tion and binding of the target will be discussed below. In addition to
the ligand, some of the block copolymers also contain functional
groups that can be used to crosslink the micelles. An ideal block
copolymer-based drug delivery system would probably combine
core or shell crosslinking with the presence of ligands at its periphery.
Only if a micelle is stable enough to retain all its drug until the
target is reached and is able to release the drug only at the desired
target in vivo, then: (1) the effect of the drug can be maximal and
(2) the side-effects of the drug, resulting from undesired release at
non-targets, can be avoided or minimized.
The aldehyde functionalized PLA–PEG diblock copolymers discussed
in Section 2.1 do not only allow attachment of the crosslinked micelles
to appropriate surfaces, but also enable the introduction of peptide
ligands through Schiff base formation and subsequent reductive
amination [34]. Functionalization of block copolymer micelles with
(oligo)peptides is not only of interest for the development of targeted
drug delivery systems, but could also be used to study the effect of
surface charges on the pharmacokinetic behaviour of the micelles.
Saccharides play an important role in cell–protein and cell–cell
interactions and therefore are another class of ligands that are of
great interest to achieve active drug targeting. Kataoka et al. prepared
galactose and glucose functionalized PLA–PEG block copolymers
through the ring-opening polymerization of successively ethylene
oxide and D,L-lactide, using protected sugars as the initiator [35].
The binding ability of the sugar-functionalized micelles was con-
firmed by means of affinity chromatography, using a RCA-1 lectin
functionalized column. RCA-1 is a plant lectin well known to bind
β-D-galactose residues. As shown in Fig. 5 the unsubstituted and glu-
cose functionalized micelles were not recognized by the lectin and
were rapidly eluted, giving a sharp, single peak in the chromatogram.
The galactose-bearing micelles, in contrast, were found to interact
strongly with the column, indicating recognition and binding by
RCA-1.
Similarly, Yonese et al. prepared sugar-substituted poly(γ-
methylglutamate)-b-poly(ethylene oxide) (PMG–PEO) block co-
polymers via ring-opening polymerization of L-glutamic acid γ-
methylester N-carboxy anhydride [36]. The polymerization was
performed with a heterobifunctional initiator, which contained
a primary amine group to start the ring-opening polymeriza-
tion at one terminus and a lactose residue to allow targeted
drug delivery at the other chain end. Recognition and binding
of the sugar ligands was evidenced by addition of RCA120 lectin
to a micellar solution of the block copolymers, which lead to a
rapid increase in the turbidity of the solution. This binding pro-
cess could be reversed by adding a large excess of lactose to
the solution containing the lectin–micelle complexes.
3. Auxiliary agents
An alternative to chemical modifications is the application of aux-
iliary agents to improve the performance of block copolymer-based
drug delivery devices. Auxiliary agents, which can be organic, inor-
ganic, high molecular weight or low molecular weight, are incorpo-
rated in the block copolymer micelle and can be located in the core
and/or in the corona. Auxiliary agents can be chosen which affect
either the properties (e.g. stability, permeability) of the micellar car-
rier or which can influence the characteristics of the encapsulated
drug (e.g. by transforming it from a lipophilic precursor into the
 A. Rösler et al. / Advanced Drug Delivery Reviews 64 (2012) 270–279
Fig. 7. (a) IR light-induced thermal activation of Au nanoparticles can cause collapse of acrylamide-based hydrogels. (b) Repetitive exposure of a hydrogel to IR laser light can be
used to create multiple bursts of encapsulated molecules.
active hydrophilic drug under a certain set of conditions). Auxiliary
agents are generally used to improve temporal control over the
drug release process, for example by allowing pulsatile drug release
under the influence of appropriate external stimuli. Below, we will
discuss two examples of auxiliary agents which have already been
successfully used, or, which we believe, could also have great poten-
tial for the advancement of block copolymer-based drug delivery
devices.
3.1. Channel proteins
In some very elegant recent work, Meier and coworkers demon-
strated that channel proteins can be incorporated without loss of activ-
ity within the hydrophobic domain of vesicles or membranes obtained
via self-assembly of amphiphilic triblock copolymers [37–39]. In their
experiments, Meier et al. focused on the so-called outer membrane pro-
tein OmpF, which occurs in the outer cell wall of Gram-negative bacte-
ria. The porin OmpF has at least two properties that make it an attractive
candidate for drug release applications. First of all, OmpF can act as a
size-selective filter, only allowing passive diffusion of small solutes,
such as ions or antibiotics. Species with a molecular weight above
400 Da are sterically excluded. This is attractive for drug delivery pur-
poses, since drugs encapsulated in a nanocage functionalized with
OmpF would be effectively protected from premature degradation by
enzymes. The drug molecules, however, would be able to diffuse freely
277
through the channel, driven by the concentration difference between
the interior and the exterior of the nanocage. The second attractive
feature of OmpF is that the channels can be closed by applying a suffi-
cient Donnan potential across the membrane it is incorporated in [40].
Upon removal of the Donnan potential, for example by the addition of
a competing low molecular weight electrolyte or by simple dilution,
the OmpF channels can be opened again. For drug delivery applications
this may allow the use of external stimuli to trigger the release of mul-
tiple bursts of drugs from nanocages functionalized with channel
proteins.
The experiments reported by Meier et al. are shown schematically in
Fig. 6. To demonstrate the feasibility of channel proteins such as OmpF
for the preparation of stimuli-sensitive nanocontainers, amphiphilic
poly(2-methyloxazoline)-b-poly(dimethylsiloxane)-b-poly(2-methyl-
oxazoline) (PMOXA–PDMS–PMOXA) triblock copolymers were used.
Mixing a solution of this triblock copolymer with a solution of the chan-
nel protein and the enzyme β-lactamase resulted in vesicles with the
OmpF protein bridging the hydrophobic PDMS shell and the enzyme
encapsulated in the nanocontainer. If desired, the nanocontainers
could be stabilized by photopolymerization of methacrylate groups
present at the terminus of the hydrophilic oxazoline blocks [41]. The
enzyme β-lactamase was encapsulated in the nanocontainers to dem-
onstrate that OmpF can be reversibly activated and deactivated with
external stimuli. Under normal circumstances, the OmpF channels are
open and substrates for the enzyme such as ampicillin can freely diffuse
278 A. Rösler et al. / Advanced Drug Delivery Reviews 64 (2012) 270–279
into the nanocontainer where it is hydrolyzed into ampicillinoic acid.
This reaction product can be easily detected when it is released from
the nanocontainer. The addition of a polyelectrolyte to the aqueous so-
lution containing the vesicles, however, creates a Donnan potential and
results in closure of the OmpF channels. In this case, no ampicillinoic
acid could be detected, since transport of substrate into and release of
product from the nanocontainers was inhibited. This blockade could
be released by addition of a competitive low molecular mass electrolyte
or by simply diluting the aqueous solution.
3.2. Metal (nano)particles
The use of thermosensitive micelles to trigger a burst-like drug
release upon local hyper- or hypothermia was mentioned in the
Introduction. This strategy to improve temporal control over the
drug release process relies on the LCST behavior of one of the seg-
ments of the block copolymer that constitutes the micelle. Although
this method is clinically feasible it suffers from some restrictions. A
major disadvantage is that individual micelles cannot be addressed
and the phase transition of the thermosensitive block copolymer
can only be induced indirectly by locally raising or lowering the tem-
perature of the environment. As a result, only those thermosensitive
micelles located closely underneath the skin may be activated, and
it might become progressively difficult to trigger micelles that are
present deeper in the body, for example in the bloodstream.
A strategy that could allow thermosensitive micelles to be individ-
ually addressed and which may also pose less restrictions on the loca-
tion of the micelles is the use of metal or metaloxide (nano)particles
as auxiliary agents. Au and γ-Fe2O3 particles, for example, can be
thermally activated by irradiation with IR light or by exposure to
an alternating magnetic field, respectively [42,43]. Since IR light
can pass largely unabsorbed through tissue [44] and also exposure
to magnetic fields does not result in unspecific heating, metal or
metaloxide particles appear to be very attractive auxiliary agents to
allow individual activation of thermosensitive micelles.
The feasibility of Au and γ-Fe2O3 particles to trigger photo- or mag-
netic field-induced phase transitions in thermosensitive polymers has
been demonstrated by West et al. [42] and Takahashi and coworkers
[43], respectively. These authors did not investigate thermosensitive
micelles, but focused on hydrogels composed of N-isopropylacrylamide
and acrylamide. Exposure to an alternating magnetic field was found to
result in an almost 10 °C increase in temperature of γ-Fe2O3-containing
hydrogels. Raising the temperature above the LCST results in a collapse
of the hydrogel, and initiates a burst-like release of encapsulated material.
This is illustrated in Fig. 7, which shows that repetitive exposure to IR
irradiation over a short period can be used to create multiple bursts of
bovine serum albumin (BSA) from an acrylamide-based hydrogel. The
ability to trigger pulsatile release is of particular interest for the delivery
of insulin in the treatment of diabetes mellitus.
Although metal particles have only been applied as auxiliary agents
to improve the temporal release properties of hydrogels up to now,
we believe that they also offer great prospects for the development
of advanced drug delivery devices based on thermosensitive block
copolymer micelles.
4. Conclusions and outlook
Over the past decade or so, amphiphilic diblock copolymer
micelles have established themselves as effective carriers for drug
delivery applications. In this review, we have discussed two different
approaches that can be used to improve the effectiveness of such block
copolymer-based drug delivery devices. The first approach involves the
chemical modification of the amphiphilic block copolymer building
blocks. Generally, the goal of these modifications is to introduce
crosslinkable groups at one of the blocks or to substitute the hydrophilic
terminus of the block copolymer with a specific ligand. Crosslinking,
either of the core or of the corona, can increase the stability of the
micelles, resulting in increased circulation times and a more gradual
release profile. The introduction of crosslinkable groups at the micellar
corona offers the additional possibility to regulate the rate of release
via the crosslink density of the corona. Thus, the deliberate introduction
of specific amounts of crosslinkable groups presents various opportuni-
ties to improve temporal control of the drug release process. Special
peptide or saccharide ligands attached to the hydrophilic chain end of
an amphiphilic block copolymer, in contrast, can contribute to improv-
ing distribution control by mediating recognition and binding of the
drug carrier to the specific site of interest.
The second approach to improve the performance of block
copolymer-based drug delivery devices is the addition of auxiliary
agents. As typical examples, channel proteins and metal (nano)particles
have been discussed. A very attractive feature of this approach is that it
does not require chemical modification of the block copolymer, since
the auxiliary agents are simply physically entrapped in the micelles.
Application of such auxiliary agents can improve temporal control by
allowing pulsatile drug release under the influence of external stimuli
such as, for example, IR light or magnetic fields.
The examples discussed throughout this review clearly underline
that significant advances can be made in the effectiveness of block
copolymer-based drug delivery devices, both with respect to temporal
as well as to distribution control over the release process. So far, most
of the work that has been published has focused on only one particular
aspect, for example on the introduction of crosslinkable groups to
increase the micellar stability, on the introduction of special ligands to
allow active targeting or on the incorporation of auxiliary agents to
enable pulsatile release. A future challenge would be to combine all of
these features into one micellar drug carrier whose mode of operation
can be adjusted to the particular needs. Furthermore, several concepts
have been introduced in this review which are relatively new and
whose potential with respect to drug delivery applications has not
yet been fully explored. This holds particularly for the core-crosslinked
micelles and the corresponding nanocontainers and also for the applica-
tion of auxiliary agents. It will be very exciting to see how these recent
developments will affect future drug delivery systems.
Acknowledgements
The authors are grateful to Prof. Klaus Müllen for his continued in-
terest and support of this work. Financial support from the Deutsche
Forschungsgemeinschaft (DFG, Emmy Noether Program, KL1049/2-1),
the Fonds der Chemischen Industrie and the Bundesministerium für
Bildung und Forschung (BMBF) is gratefully acknowledged.
References
[1] G.S. Kwon, K. Kataoka, Block copolymer micelles as long-circulating drug vehicles,
Adv. Drug Deliv. Rev. 16 (1995) 295–309.
[2] G.S. Kwon, T. Okano, Polymeric micelles as new drug carriers, Adv. Drug Deliv.
Rev. 21 (1996) 107–116.
[3] G.S. Kwon, Diblock copolymer nanoparticles for drug delivery, Crit. Rev. Ther.
Drug 15 (1998) 481–512.
[4] L. Yang, P. Alexandridis, Physicochemical aspects of drug delivery and release
from polymer-based colloids, Curr. Opin. Colloid In. 5 (2000) 132–143.
[5] C. Allen, D. Maysinger, A. Eisenberg, Nano-engineering block copolymer aggregates
for drug delivery, Colloids Surfaces B 16 (1999) 3–27.
[6] K.E. Uhrich, S.M. Cannizzaro, R.S. Langer, K.M. Shakesheff, Chem. Rev. 99 (1999)
3181–3198.
[7] S. Cammas-Marion, T. Okano, K. Kataoka, Functional and site-specific macromolecular
micelles as high potential drug carriers, Colloids Surfaces B 16 (1999) 207–215.
[8] J.E. Chung, M. Yokoyama, T. Okano, Inner core segment design for drug delivery con-
trol of thermo-responsive polymeric micelles, J. Control. Release 65 (2000) 93–103.
[9] M.D.C. Topp, P.J. Dijkstra, H. Talsma, J. Feijen, Thermosensitive micelle-forming
block copolymers of poly(ethylene glycol) and poly(N-isopropylacrylamide),
Macromolecules 30 (1997) 8518–8520.
[10] M. Yokoyama, T. Okano, Y. Sakurai, S. Fukushima, K. Okamoto, K. Kataoka, Selective
delivery of adriamycin to a solid tumor using a polymeric micelle carrier system,
J. Drug Target. 7 (1999) 171–186.
 A. Rösler et al. / Advanced Drug Delivery Reviews 64 (2012) 270–279
[11] H. Maeda, J. Wu, T. Sawa, Y. Matsumura, K. Hori, Tumor vascular permeability and
the EPR effect in macromolecular therapeutics: a review, J. Control. Release 65
(2000) 271–284.
[12] Y. Matsumura, H. Maeda, A new concept for macromolecular therapeutics in
cancer chemotherapy: mechanism of tumoritropic accumulation of proteins and
the antitumor agent smancs, Cancer Res. 46 (1986) 6378–6392.
[13] M. Iijima, Y. Nagasaki, T. Okada, M. Kato, K. Kataoka, Core-polymerized reactive
micelles from heterotelechelic amphiphilic block copolymers, Macromolecules
32 (1999) 1140–1146.
[14] J.-H. Kim, K. Emoto, M. Iijima, Y. Nagasaki, T. Aoyagi, T. Okano, Y. Sakurai, K.
Kataoka, Core-stabilized polymeric micelle as potential drug carrier: increased
solubilization of taxol, Polym. Adv. Technol. 10 (1999) 647–664.
[15] K. Emoto, Y. Nagasaki, K. Kataoka, Coating of surfaces with stabilized reactive micelles
from poly(ethylene glycol)–poly(D,L-lactic acid) block copolymer, Langmuir 15
(1999) 5212–5218.
[16] K. Emoto, Y. Nagasaki, K. Kataoka, A core-shell structured hydrogel thin layer on
surfaces by lamination of a poly(ethylene glycol)-b-poly(D,L-lactide) micelle and
polyallylamine, Langmuir 16 (2000) 5738–5742.
[17] K. Emoto, M. Iijima, Y. Nagasaki, K. Kataoka, Functionality of polymeric micelle
hydrogels with organized three-dimensional architecture on surfaces, J. Am.
Chem. Soc. 122 (2000) 2653–2654.
[18] Y.-Y. Won, H.T. Davis, F.S. Bates, Giant wormlike rubber micelles, Science 283
(1999) 960–963.
[19] F. Henselwood, G. Liu, Water-soluble nanospheres of poly(2-cinnamoylethyl
methacrylate)-block-poly(acrylic acid), Macromolecules 30 (1997) 488–493.
[20] Y. Kakizawa, A. Harada, K. Kataoka, Environment-sensitive stabilisation of
core-shell structured polyion complex micelle by reversible cross-linking of the
core through disulfide bond, J. Am. Chem. Soc. 121 (1999) 11247–11248.
[21] Y. Kakizawa, A. Harada, K. Kataoka, Glutathione-sensitive stabilisation of block
copolymer micelles composed of antisense DNA and thiolated poly(ethylene
glycol)-block-poly(L-lysine): a potential carrier for systemic delivery of antisense
DNA, Biomacromolecules 2 (2001) 491–497.
[22] K.B. Thurmond, T. Kowalewski, K.L. Wooley, Water-soluble knedle-like structures:
the preparation of shell-cross-linked small particles, J. Am. Chem. Soc. 118 (1996)
7239–7240.
[23] K.S. Murthy, Q. Ma, C.G. Clark Jr., E.E. Remsen, K.L. Wooley, Fundamental design
aspects of amphiphilic shell-crosslinked nanoparticles for controlled release
applications, Chem. Commun. (2001) 773–774.
[24] H. Huang, E.E. Remsen, T. Kowalewski, K.L. Wooley, Nanocages derived from shell
cross-linked micelle templates, J. Am. Chem. Soc. 121 (1999) 3805–3806.
[25] T. Sanji, Y. Nakatsuka, S. Ohnishi, H. Sakurai, Preparation of nanometer-sized
hollow particles by photochemical degradation of polysilane shell cross-linked
micelles and reversible encapsulation of guest molecules, Macromolecules 33
(2000) 8524–8526.
[26] Q. Zhang, K.L. Wooley, Synthesis and characterization of shell cross-linked
nanoparticles containing a degradable core domain, Polym. Prepr. 42 (2) (1999)
986–987.
279
[27] V. Bütün, N.C. Billingham, S.P. Armes, Synthesis of shell cross-linked micelles with
tunable hydrophilic/hydrophobic cores, J. Am. Chem. Soc. 120 (1998) 12135–12136.
[28] V. Bütün, A.B. Lowe, N.C. Billingham, S.P. Armes, Synthesis of zwitterionic shell-
crosslinked micelles, J. Am. Chem. Soc. 121 (1999) 4288–4289.
[29] J. Ding, G. Liu, Polystyrene-b-poly(2-cinnamoylethyl methacrylate) nanospheres
with cross-linked shells, Macromolecules 31 (1998) 6554–6558.
[30] S. Stewart, G. Liu, Hollow nanospheres from polyisoprene-block-poly(2-cinnamoylethyl
methacrylate)-block-poly(tert-butyl acrylate), Chem. Mater. 11 (1999) 1048–1054.
[31] J. Ding, G. Liu, Water-soluble hollow nanospheres as potential drug carriers,
J. Phys. Chem. B 102 (1998) 6107–6113.
[32] K.L. Wooley, From dendrimers to knedel-like structures, Chem. Eur. J. 3 (9) (1997)
1397–1399.
[33] J. Liu, K.L. Wooley, Internal and external surface functionalization of a shell
crosslinked nanocage: potential for drug delivery by bioconjugates of polymeric
nanomaterials, Polym. Mater. Sci. 84 (2001) 967–968.
[34] Y. Yamamoto, Y. Nagasaki, M. Kato, K. Kataoka, Surface charge modulation of
poly(ethylene glycol)–poly(D,L-lactide) block copolymer micelles: conjugation
of charged peptides, Colloids Surfaces B 16 (1999) 135–146.
[35] K. Yasugi, T. Nakamura, Y. Nagasaki, M. Kato, K. Kataoka, Sugar-installed polymer
micelles: synthesis and micellization of poly(ethylene glycol)–poly(D,L-lactide)
block copolymers having sugar groups at the PEG chain end, Macromolecules
32 (1999) 8024–8032.
[36] A. Toyotama, S. Kugimiya, J. Yamanaka, M. Yonese, Preparation of a novel aggregate
like sugar-ball micelle composed of poly(methylglutamate) and poly(ethyleneglycol)
modified by lactose and its molecular recognition by lectin, Chem. Pharm. Bull. 49
(2001) 169–172.
[37] C. Nardin, J. Widmer, M. Winterhalter, W. Meier, Amphiphilic block copolymer
nanocontainers as bioreactors, Eur. Phys. J. E 4 (2001) 403–410.
[38] C. Nardin, S. Thoeni, J. Widmer, M. Winterhalter, W. Meier, Nanoreactors based on
(polymerized) ABA-triblock copolymer vesicles, Chem. Commun. (2000) 1433–1434.
[39] W. Meier, C. Nardin, M. Winterhalter, Reconstitution of channel proteins in
(polymerized) ABA triblock copolymer membranes, Angew. Chem. Int. Ed. 39
(2000) 4599–4602.
[40] P. Van Gelder, F. Dumas, M. Winterhalter, Understanding the function of bacterial
outer membrane channels by reconstitution into black lipid membranes, Biophys.
Chem. 85 (2000) 153–167.
[41] C. Nardin, Th. Hirt, J. Leukel, W. Meier, Polymerized ABA triblock copolymer
vesicles, Langmuir 16 (2000) 1035–1041.
[42] S.R. Sershen, S.L. Westcott, N.J. Halas, J.L. West, Temperature-sensitive polymer-
nanoshell composites for photothermally modulated drug delivery, J. Biomed.
Mater. Res. 51 (2000) 293–296.
[43] N. Kato, A. Oishi, F. Takahashi, Enzyme reaction controlled by magnetic heating
due to the hysteresis loss of γ-Fe2O3 in thermosensitive polymer gels immobilized
β-galactosidase, Mater. Sci. Eng. C: Biol. 6 (1998) 291–296.
[44] C.R. Simpson, M. Kohl, M. Essenpreis, M. Cope, Near-infrared optical properties of
ex-vivo human skin and subcutaneous tissues measured using the Monte Carlo
inversion technique, Phys. Med. Biol. 43 (1998) 2465–2478.