Advanced Review

Synthesis of poly(alkyl
cyanoacrylate)-based colloidal
nanomedicines
Julien Nicolas ∗ and Patrick Couvreur1
Nanoparticles developed from poly(alkyl cyanoacrylate) (PACA) biodegradable
polymers have opened new and exciting perspectives in the field of drug delivery
due to their nearly ideal characteristics as drug carriers in connection with
biomedical applications. Thanks to the direct implication of organic chemistry,
polymer science and physicochemistry, multiple PACA nanoparticles with
different features can be obtained: nanospheres and nanocapsules (either oil- or
water-containing) as well as long-circulating and ligand-decorated nanoparticles.
This review aims at emphasizing the synthetic standpoint of all these nanoparticles
by describing the important aspects of alkyl cyanoacrylate chemistry as well as the
experimental procedures and the different techniques involved for the preparation
of the corresponding colloidal devices.
 2008 John Wiley & Sons, Inc. Wiley Interdiscipl. Rev. Nanomed. Nanobiotechnol. 2009 1 111–127

N anotechnology has emerged as a promising

area of research in which scientists from both
academia and industry put a lot of effort, hoping
Introduced more than 25 years ago in the field of
pharmacology, PACA drug carriers have demon-
strated significant results in numerous pathologies
for the best with regard to life in future. It is such as cancer, severe infections (viral, bacteriologic,
a highly multidisciplinary field which consists of parasite) as well as several metabolic and autoimmune
engineering functional systems at the molecular scale diseases, well-reviewed in the recent literature.1–6 As
and covers applied physics, materials science, interface a complementary work, the objective of the present
and colloid science, supramolecular chemistry as well review is to emphasize the synthetic aspect of these
as chemical, mechanical, and electrical engineering. colloidal carriers by describing, as precisely as pos-
One of the direct applications of nanotechnology is sible, the chemistry of the cyanoacrylate monomers,
devoted to the medical and pharmacology areas, also their polymerization as well as the different structures
called nanomedicine, the most famous example being and morphologies of the corresponding nanoparticles.
nanoparticle drug delivery. In particular, description of this PACA-based nan-
Indeed, a crucial impulse was given to otechnology will start from the simplest nanocarriers
nanomedicine with the development of various to more sophisticated and ‘smart’ drug delivery
types of drug-carrier nanodevices, made possible by devices. The reader who would like a more exhaustive
means of multidisciplinary approaches–organic and point of view about the biologic and pharmaceutical
polymer chemistry, physicochemistry, pharmacology, aspects of PACA nanoparticles as well as the drugs
etc. Among suitable nanodevices for drug delivery, successfully incorporated in such colloidal devices is
nanoparticles on the basis of biodegradable poly(alkyl referred to the above-mentioned references.
cyanoacrylate) (PACA) polymers have appeared as
an established technology for colloidal nanomedicine.
ALKYL CYANOACRYLATE
∗ Correspondence to: Julien Nicolas, Laboratoire de Physico- MONOMERS AND RELATED
Chimie, Pharmacotechnie et Biopharmacie, UMR CNRS
8612, Univ Paris-Sud, 92296 Châtenay Malabry, France. POLYMERS
E-mail: [email protected]
1
General Features of Alkyl Cyanoacrylate
Laboratoire de Physico-Chimie, Pharmacotechnie et Biopharmacie,
UMR CNRS 8612, Univ Paris-Sud, 92296 Châtenay Malabry, Monomers
France. Alkyl cyanoacrylates are widely known monomers,
DOI: 10.1002/wnan.015 extremely appreciated for their very high reactivity

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 Advanced Review www.wiley.com/wires/nanomed

and the excellent adhesive properties of the resulting CN

(a) CN − H2O
polymers. On one hand, the famous Superglue n + n CH2O
n
(manufactured by Henkel), which contains short COOR
O O
alkyl chain cyanoacrylates, is commonly employed R
by the general public for repairing and do-it-yourself
activities, whereas longer alkyl chain cyanoacrylates (b) CN CN
have been developed for biomedical purposes such as ∆
n
surgical glue for the closure of skin wounds7–13 and n O
embolitic material for endovascular surgery.10,11,14 O O O
R R
Indeed, several commercial products have emerged
from the use of cyanoacrylates in the biomedical FIGURE 2 | Synthesis of alkyl cyanoacrylate monomer via
area, mainly devoted to tissue adhesion. For instance, Knoevenagel condensation reaction (a) and subsequent thermal
methyl cyanoacrylate (MCA, Figure 1) is the main depolymerization (b).
component of the Biobond tissue adhesive and longer
alkyl ester chain cyanoacrylates, such as n-butyl
cyanoacrylate (nBCA, Figure 1) or octyl cyanoacrylate with small amounts of a free-radical inhibitors to
(OCA, Figure 1), were commercialized under the prevent repolymerization (Figure 2).
trademarks of Indermil, Liquidband, and Dermabond, From that moment on, the synthetic protocol
respectively. remained almost unchanged. It was only slightly mod-
The synthesis of alkyl cyanoacrylate monomers ified and improved essentially by playing with the
has been described in the patent literature since nature of the solvent mixture,19,20 by applying a
1949.15–18 Basically, the main strategy to achieve transesterification approach for making cyanoacry-
α-cyanoacrylates comprises two steps. First, the lates bearing longer alkyl ester chains,21 or by using
corresponding alkyl cyanoacetate is reacted with a more efficient catalyst (namely pyrrolidine) for the
formaldehyde in the presence of a basic catalyst, to condensation step.22
form PACA oligomers (by the so-called Knoevenagel
condensation reaction). The catalyst is a base, either
inorganic (e.g., sodium or potassium hydroxide, Polymerization of Alkyl Cyanoacrylates
ammonia) or organic (e.g., quilonine, piperidine, in Homogeneous Media
dialkyl amines). Then, pure alkyl cyanoacrylate On the fringe of typical vinyl monomers [styrenics,
monomer is recovered by a thermal depolymerization (meth)acrylates, etc.] is the alkyl cyanoacrylate family,
reaction of the previously obtained oligomers, using which seems to be an exotic class of polymerizable
suitable stabilizers such as protonic or Lewis acids compounds. Indeed, due to the presence of two
powerful electro-withdrawing groups in the α-carbon
CN CN CN CN of the double bond, namely ester (COOR) and
O O O O cyano (CN), alkyl cyanoacrylate monomers exhibit
O O O O a remarkable reactivity toward nucleophiles such
as anions (hydroxide, iodide, alcoholate, etc.) or
weak bases (alcohol, amine, etc.), resulting in a very
MCA ECA nBCA IBCA high polymerization rate. Even traces of one of the
CN CN CN CN CN
above-mentioned compounds in the reaction medium
are sufficient to initiate such a fast polymerization.
O O O O O
O O O O O
This explains why alkyl cyanoacrylates are extremely
difficult to handle under their pure form and that
4 13 14 O
batches of these monomers are usually maintain stable
n with a small amount of acidic stabilizers (e.g., SO2 ,
IHCA OCA ISCA HDCA MePEGCA sulfonic acid, etc.).
PACA can be synthesized according to
FIGURE 1 | Structure of alkyl cyanoacrylates described in the three distinct types of polymerization: (1) anionic,
literature: methyl cyanoacrylate (MCA), ethyl cyanoacrylate (ECA), (2) zwitterionic, and (3) radical (Figure 3). In prac-
n-butyl cyanoacrylate (nBCA), isobutyl cyanoacrylate (IBCA), isohexyl tice, because of the exceptional reactivity of alkyl
cyanoacrylate (IHCA), octyl cyanoacrylate (OCA), isostearyl cyanoacrylate derivatives, anionic and zwitterionic
cyanoacrylate (ISCA), hexadecyl cyanoacrylate (HDCA), and
polymerization mechanisms are by far predominant
methoxypoly(ethylene glycol) cyanoacrylate (MePEGCA).
under conventional experimental conditions with

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 WIREs Nanomedicine and Nanobiotechnology Synthesis of PACA-based colloidal nanomedicines

(a)
CN B CN
– –
B
O O CN
CN
O O B
R R n
O
O O O
R R
(b)
CN Nu CN
Nu –
O O CN
CN
O O Nu
R R n
O
O O O
R R
(c)
CN P CN
P
O O CN
CN
O O P
R R n
O
O O O
R R

FIGURE 3 | Initiation and propagation steps involved during anionic (a), zwitterionic (b), and radical (c) polymerizations of alkyl cyanoacrylate
monomer initiated by a base (B− ), a nucleophile (Nu), and a radical (P• ), respectively.

respect to a pure radical process. This explains why acetates) as the initiating species for the polymeriza-
studies on alkyl cyanoacrylates polymerization in both tion of nBCA at 20–40◦ C in tetrahydrofuran (THF)
homogeneous (i.e., bulk or solution) and hetero- and reported a nearly ideal living polymerization in
geneous (i.e., emulsion, microemulsion) media were the case of the hydroxide-based initiator.31–33
mainly devoted to anionic and zwitterionic processes. Even though anionic and zwitterionic mecha-
nisms are more likely to occur for the polymerization
Synthesis of Homopolymers of alkyl cyanoacrylates, free-radical polymerization
In this field, an extensive work has been accom- was believed to be the main chain-extension
plished by Pepper and coworkers to get a better process during homopolymerization34–37 and
understanding of the involved polymerization mecha- copolymerization37,38 carried out in bulk when a suit-
nisms depending on the experimental conditions.23–26 able inhibitor is introduced in the reaction medium.
Indeed, the homopolymerization in solution of ethyl However, even under these specific inhibition condi-
cyanoacrylate (ECA, Figure 1) and nBCA were initi- tions, anionic polymerization is not totally suppressed
ated either by simple anions (CH3 COO− , CN− , I− , but is made negligible regarding the timescale of the
etc.) or by covalent organic bases (Et3 N, pyridine, polymerization reaction. In particular, Canale et al.34
etc.), leading to anionic or zwitterionic polymer- used in 1960, boron trifluoride–acetic acid complex
izations, respectively.23 For zwitterionic polymeriza- while conducting free-radical bulk polymerization
tion of nBCA, the influence of the nature of the of MCA at 60◦ C initiated by azobisisobutyronitrile
initiator as well as other experimental conditions (AIBN), whereas Bevington et al.36 used propane-
(inhibiting species, presence of water, etc.) on both 1,3-sultone as an efficient inhibitor against anionic
the main characteristics of the obtained polymer polymerization for the free-radical polymerization of
(number-average molecular weight, molecular weight MCA in bulk or in 1,4-dioxane at 60◦ C, initiated by
distribution) and polymerization kinetics (monomer AIBN or benzoylperoxide (BPO). In 1983, Yamada
conversion, polymerization rate, etc.) were investi- et al.37 polymerized ECA in bulk at 30◦ C with a
gated through a small library of covalent organic bases small amount of acetic acid or propane-1,3-sultone
such as phosphine,26–29 pyridine24,27 and amine25,27,30 and from their results, they extracted very high
derivatives. Considering anionic polymerization, the propagation rate constants: kp = 1622 l mol−1 s−1
same research group used tetrabutyl ammonium in the presence of acetic acid and kp = 1610 l mol−1
salts (hydroxide, bromide, acetate, and substituted s−1 in the presence of propane-1,3-sultone. As a

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comparison, methyl methacrylate (MMA) which CN CN CN CN

is considered as a highly reactive monomer gave Me2NH
O + O + CH2O
kp = 450 l mol−1 s−1 at 30◦ C.39
x y
O O EtOH, 20 °C
O O O O

14 O
Synthesis of Copolymers n
14 O
n
Alkyl cyanoacrylates were also copolymerized with
more ‘common’ vinyl monomers through a free- FIGURE 4 | Synthesis of random, comb-like poly[(hexadecyl
radical process (using trifluoride–acetic acid complex cyanoacrylate)-co-methoxypoly(ethylene glycol) cyanoacrylate]
as an efficient inhibitor against anionic polymeriza- [P(HDCA-co-MePEGCA)] copolymer via Knoevenagel condensation
tion) to give different kinds of copolymers, depending reaction.
on the nature of the comonomer.38 Random copoly-
mers with MMA were achieved in bulk, whereas diblock and PIBCA-b-PEG-b-PIBCA triblock copoly-
alternating copolymers with styrene were reported in mers with tuneable compositions in good match with
benzene solution at 60◦ C under AIBN initiation. Con- the initial stoichiometry.
sidering bulk properties, random copolymers with Synthesis of poly[(hexadecyl cyanoacrylate)-
10% MMA exhibit physical properties similar to co-methoxypoly(ethylene glycol) cyanoacrylate]
the PMCA homopolymer, whereas alternating copoly- [P(HDCA-co-MePEGCA)] comb-like copolymers
mers with styrene had an enhanced thermal stability exhibiting amphiphilic properties was reported by
compared with random copolymers. Hall et al., who Peracchia et al.46 This original approach derived from
previously investigated the reactions of electron-rich Knoevenagel condensation reaction where corre-
olefins with electron-poor olefins,40–42 confirmed the sponding cyanoacetates, namely hexadecyl cyanoac-
alternating copolymer starting from a 1:1 styrene : etate and PEG monomethyl ether cyanoacetate, were
MCA mixture, either initiated by AIBN under UV reacted with formaldehyde in the presence of dimethy-
light at 40◦ C in benzene solution or produced spon- lamine as the catalyst (Figure 4). Thanks to the slow,
taneously at room temperature.43 However, when in situ formation of the cyanoacrylate monomers, it
using other comonomers such as isobutyl vinyl ether, allowed the polymerization process to be better con-
p-methoxystyrene or β-bromostyrene, copolymeriza- trolled compared with a direct anionic polymerization.
tions with MCA led to mixtures of (co)polymers Besides, the composition of the copolymer (and thus
and/or small adducts.43 its hydrophilicity/hydrophobicity) can be adjusted
In 1978, a comprehensive synthetic approach of simply by varying the initial cyanoacetates feed ratio.
bis(alkyl cyanoacrylate)s was proposed by Buck start-
ing from anthracene adducts.44 These difunctional POLY(ALKYL
alkyl monomers derived from cyanoacrylates were
CYANOACRYLATE)-BASED
copolymerized with monofunctional alkyl cyanoacry-
lates such as MCA and isobutyl cyanoacrylate
NANOPARTICLES
(IBCA, Figure 1), resulting in crosslinked macro- General Consideration on the Synthesis
molecular adhesive compositions exhibiting supe- of Poly(alkyl cyanoacrylate) Nanoparticles
rior mechanical properties under both dry and wet Nanoparticle is a collective name for two different
environments than the noncrosslinked counterparts, types of colloidal objects, namely nanospheres (NS)
which could be advantageously employed as pit and and nanocapsules (NC), which can be separately
fissure sealant in dentistry. obtained depending on the preparation process.
More sophisticated macromolecular architec- Basically, nanospheres are matrix systems constituted
tures such as diblock and triblock copolymers com- by the polymer in which the drug is physically
prising poly(ethylene glycol) (PEG) and PACA blocks and uniformly dispersed, whereas nanocapsules are
were also synthesized in homogeneous media via zwit- vesicular systems in which the drug is solubilized in
terionic polymerization.45 The synthesis involved the a liquid core, either water (w-NC) or oil (o-NC),
preparation of triphenylphosphine end-capped mono- surrounded by a thin polymer layer (Figure 5).
hydroxyl and dihydroxyl PEGs, giving the correspond- During the last 25 years, an important break-
ing monofunctional and difunctional macrozwitteri- through in this field has been witnessed with the
onic initiator. The polymerization of IBCA was then development of PACA nanoparticles as colloidal
initiated with each one of the macroinitiators in drug carriers. Polymerizations in heterogeneous
THF at ambient temperature to afford PIBCA-b-PEG media (i.e., emulsion, dispersion, miniemulsion,

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 WIREs Nanomedicine and Nanobiotechnology Synthesis of PACA-based colloidal nanomedicines

(a) CN – HO CN
HO
–
O O
O O
R R
(b)
HO CN CN CN CN
– + n HO –
NS w-NC o-NC O O n
O O O OO O
FIGURE 5 | Schematic representation of nanospheres (NS), R R R R
water-containing nanocapsules (w-NC), and oil-containing
nanocapsules (o-NC).
CN CN CN CN
(c) HO –
H
HO
n n H
microemulsion)47,48 and spontaneous emulsification O OO O HO
–
O OO O
techniques49–51 are two well-known approaches for R R R R
the preparation of polymeric particles, which have
also been intensively used for the confection of PACA FIGURE 6 | Schematic representation of poly(alkyl cyanoacrylate)
nanoparticles as colloidal drug carriers for in vivo formation via the stepwise anionic polymerization mechanism in
administration. emulsion/dispersion. Initiation step (a), reversible propagation step (b),
and reversible termination step (c).
Synthesis of Nanospheres
In 1979, Couvreur et al. first developed a simple terminated by the acid-inhibiting agents present in
process to directly generate stable MCA or ECA the monomer. This step is followed by a re-initiation
nanospheres, consisting of a dropwise addition reaction of terminated species by still living chains,
of the monomer into a vortexed HCl solution leading to further polymerization until a molecular
(2 < pH < 3) containing a nonionic or a macro- weight balancing is reached, similar to depolymeriza-
molecular surfactant.52 Since then, numerous studies tion/repolymerization events.66 One should be aware
aiming at establishing relevant parameters governing that in all these mechanisms, the polymerization is
the polymerization kinetics as well as the characteris- postulated to be initiated by the hydroxyl ions from
tics of the macromolecules and the nanospheres have the aqueous phase independently of other reactants
been reported. It has been shown that the nature and existing in the polymerization medium.
the concentration of the surfactant played a significant On the basis on an interfacial polymerization
role on the particle size,53–61 whereas the type of both mechanism,67,68 Limouzin et al. polymerized nBCA
the monomer and the surfactant strongly influenced in emulsion and miniemulsion in the presence
the molar mass of the obtained polymer.55–58 Besides of dodecylbenzenesulfonic acid (DBSA) acting as
the monomer concentration,53,59,60,62 the pH of the both surfactant and terminating agent (also termed
reaction medium53,55–58,60–64 and the concentration of tersurf).69 By releasing protons at the water/oil
sulfur dioxide (acting as a polymerization inhibitor)57 interface, DBSA allowed the interfacial, anionic
were also crucial parameters which strongly affected polymerization to be drastically slowed down through
the macromolecular and/or colloidal properties of a (reversible) termination reaction and to proceed
the nanospheres. The size of the colloidal objects under a fairly controlled fashion leading to stable
which can be obtained usually ranged from 50 to high solids content (∼20%) PnBCA nanospheres.
300 nm,54,59,60 which is a well-adapted window The miniemulsion technique was also used by Weiss
for colloidal drug delivery devices, especially by et al. for the preparation of PnBCA nanospheres. By
intravenous administration. varying the concentration of the surfactant (SDS),
For a more fundamental standpoint, several and by adding sodium hydroxide as the initiating
tentative mechanisms have been postulated.65,66 species, high solids content dispersions up to 10%
It has been reported that the emulsion/dispersion with average diameters ranging from 110 to 360 nm
polymerization in acidic medium is not that trivial and were obtained.70
proceeds via a stepwise, anionic mechanism compris-
ing reversible propagation and reversible termination Synthesis of Nanocapsules
steps63,64 (Figure 6). Basically, PACA oligomers are Nanocapsules are reservoir-type nanoparticles in
formed in the monomer droplets and are reversibly which drugs can be encapsulated according to their

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 Advanced Review www.wiley.com/wires/nanomed

intrinsic solubility. In other words, oil-containing as surfactants. Basically, the alkyl cyanoacrylate
nanocapsules will be able to encapsulate hydropho- monomer is added to the preformed (micro)emulsion
bic drugs, whereas hydrophilic ones will be efficiently and, in a similar way to that of oil-containing nanocap-
encapsulated into water-containing nanocapsules.71 sules, spontaneous anionic polymerization occurred
The nature of the nanocapsules (i.e., water-containing at the water/oil interface to form a thin PACA
or oil-containing) is determined by the nature of the layer surrounding an aqueous core. Depending on
dispersed phase involved in a heterogeneous poly- the nature of the surfactant and the starting system
merization process, usually emulsion or microemul- (emulsion or microemulsion), which are parameters
sion. Basically, the macromolecular shell is formed governing the surface properties of these colloidal
by the spontaneous anionic polymerization of alkyl objects, this process led to 50–350 nm diameter, stable
cyanoacrylate occurring at the interface between the nanocapsules.76–79
dispersed and the continuous phase. Historically, However, because the inverse (micro)emulsion
oil-containing nanocapsules were first developed by processes conduct to water-containing nanocapsules
Fallouh et al.49 through a simple protocol: a solu- dispersed in oil (which are suitable for oral route
tion of monomer and oil in a water-miscible solvent administration), intravenous injection cannot be
(usually ethanol) is poured into an aqueous solution directly performed with a nonaqueous dispersing
of surfactant (usually Poloxamer 188) under vigor- medium. To circumvent this limitation, a recent
ous stirring, leading to small oil/monomer droplets method aiming at transferring the nanocapsules
at the interface of which the polymerization is ini- from an oil-dispersing medium to a water-dispersing
tiated by hydroxide ions present in water. Gallardo medium was recently suggested by Couvreur and
et al.65 reported that the crucial parameters for achiev- coworkers and consisted in a centrifugation step of
ing nanocapsules lies: (1) in the diffusion behavior of the nanocapsules onto an aqueous layer.77,79
the organic solvent (acting as a monomer support) To synthesize nanocapsules with preformed
within the aqueous phase, which ultimately governs polymers, homopolymer of alkyl cyanoacrylate are
the reservoir nature of the nanoparticles, and (2) in required and synthesized separately, for instance by
the simultaneous precipitation of the polymer at the dripping the monomer in pure water, the polymer
water/oil interface (i.e., the polymer should be insol- being subsequently recovered by lyophilization. The
uble in both the aqueous and the organic phase). nanocapsules preparation method, also called interfa-
Usually, nanocapsules exhibit average diameter rang- cial deposition, consists of the addition of a solution
ing from 200 to 350 nm, the latter being governed by of the homopolymer and a small amount of oil,
several physicochemical parameters such as the nature for instance Miglyol (which will constitute the oily
and the concentration of the monomer and encapsu- core of the nanocapsules), into an aqueous phase.
lated drug, the amount of surfactant and oil as well as The oil-containing nanocapsules form instantaneously
the speed of diffusion of the organic phase within the by deposition of the homopolymer at the oil/water
aqueous phase. However, Altinbas et al. have demon- interface, which precipitate as a macromolecular
strated that when a miniemulsion is applied instead shell.71,80,81 In general, a surfactant is added in the
of an emulsion, nanocapsules of an average diameter aqueous phase to ensure colloidal stability of these
below 100 nm can be obtained.72 nanocapsules.
The main drawback often encountered in this
approach is the contamination of the nanocapsule
population by a substantial amount of nanospheres, Synthesis of Poly(alkyl cyanoacrylate)
resulting from a partial polymerization in the organic Nanoparticles with Controlled Surface
phase.65 However, it has been shown that an Properties
optimized ethanol/oil ratio,65,73 the acidification of In this topic, the major breakthrough is undoubt-
the organic phase,74 and the inhibition of the edly the grafting of PEG, a nonionic, flexible, and
polymerization in the organic phase by aprotic hydrophilic polymer, onto nanoparticles (which also
solvents75 (acetonitrile, acetone) each avoided the applies for other colloidal drug carriers such as
formation of matrix-type nanoparticles. liposomes). This approach, termed ‘PEGylation’, rep-
Water-containing nanocapsules have been devel- resented a milestone in the drug delivery area.82,83
oped more recently than were the oil-containing Indeed, non-‘PEGylated’ nanoparticles are quickly
counterparts. They are usually prepared by water eliminated from the bloodstream because of the
in oil (w/o) (micro)emulsion, also called an inverse adsorption of blood proteins (opsonins) onto their
(micro)emulsion, using polysorbate, sorbit monoleate surface, which triggers the recognition of the mononu-
or poly(ethylene oxide) lauryl ester (Brij 35) clear phagocyte system (MPS) by the macrophages.

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 WIREs Nanomedicine and Nanobiotechnology Synthesis of PACA-based colloidal nanomedicines

As a consequence, these nanoparticles are ineluctably of surface-active macromolecules. This point is of

accumulated in MPS organs such as the liver and great importance since nanoparticles with covalently
the spleen, restricting the therapeutic activity of the anchored stabilizing moieties at their surface would
entrapped compounds to liver diseases (i.e., hepatic behave differently in a biologic medium than those
primary hepatocarcinoma or metastasis as well as liver with adsorbed surfactants. As a consequence, this
intracellular infections). In contrast, when covered by is only later on that researches have been strictly
PEG chains, the obtained nanoparticles are able to devoted to surface engineering of PACA nanoparticles
efficiently escape this recognition system, resulting in in order to investigate any subtle change of the surface
long-circulating, colloidal devices, also called ‘stealth’ properties of the nanoparticles on their in vivo fate.
nanoparticles.82,83 However, almost unmarked, early works
After it has been demonstrated that PACA by Douglas et al. postulated that dextran or
nanoparticles can be seen as very promising β-cyclodextrin may also initiate the polymerization of
biodegradable drug carriers (the BioAlliance Pharma butyl cyanoacrylate (BCA) resulting in the formation
spin-off company is now producing doxorubicin- of amphiphilic copolymers, helping to stabilize the
loaded PACA nanoparticles for clinical uses in nascent nanoparticles.54 This approach was revisited
phase II/III trials with resistant liver hepatocarcinoma by Peracchia et al. using different linear PEGs acting
as main indication), their complexity was further as stabilizers and initiators for the emulsion poly-
increased by performing appropriate tuning of their merization of IBCA85,86 (Figure 7(a) and (b)). It was
surface properties in order to control their in vivo fate.
demonstrated that PEG chains exhibited different con-
formations at the surface of the nanospheres: (1) hairy
Surface Modification of Nanospheres nanospheres with PEG monomethyl ether due to a
First attempts concerning surface modification of single initiation site (Figure 7(a)) or (2) long loops
PACA nanospheres logically concerned the ‘PEGy- using PEG due to the divergent chain growth (two
lation’ concept, either via a simple adsorption of PEG initiating sites) during the polymerization of IBCA87
chains onto the nanoparticles or by a covalent linkage (Figure 7(b)). In the same spirit, the use of polysac-
of PEG chains with PACA polymers. However, the charides, such as dextran, dextran sulfate, chitosan,
adsorption approach does not fit the covalent linkage and thiolated chitosan, as stabilizing/initiating agents
criteria and is not really suitable as long as it has been under similar experimental conditions also led to sta-
demonstrated that these kinds of assemblies (PACA
ble nanospheres in the 100–500 nm range, exhibiting
nanoparticles on which poloxamer 388 or poloxam-
different surface properties; for instance, positively
ine 908 was adsorbed) are not stable during in vivo
charged with chitosan61,88,89 and from rather neutral
administration, resulting in a loss of coating and no
to negatively charged with dextran derivatives.88,90,91
significant influence on the biodistribution pattern.84
So far, anionic (mini)emulsion polymerization
Thus the covalent bond of the PEG chains at the sur-
was the most widespread and straightforward tech-
face of the nanoparticles is a prerequisite for this kind
nique to synthesize PACA nanospheres. Even though,
of application.
Basically, different types of hydrophilic in that case, the mechanism is on the basis of
molecules have been anchored, on purpose, to the anionic propagating species,63,64 Chauvierre et al.
surface of PACA nanoparticles (Figure 7). Efficient recently adapted Couvreur’s original protocol to a
surface modification of nanospheres can be achieved free-radical emulsion polymerization process, thanks
either in situ during the polymerization in aqueous to the polysaccharide/cerium IV (Ce4+ ) ions redox
dispersed media or from preformed amphiphilic couple as the initiator92 (Figure 8). Because of the fast
copolymers during emulsification processes. radical initiation rate, anionic polymerization is negli-
Concerning previous studies about anionic/zwit gible regarding the timescale of the experiment which
terionic emulsion polymerization of alkyl cyanoacry- makes way for a free-radical chain growth process.
late, the hydrophilic molecules introduced in the This technique was also employed for the emulsion
recipes (SDS, dextran, poloxamer, Tweens, cyclodex- polymerization of alkyl cyanoacrylate using different
trins, etc.) were solely used as stabilizing agents for kinds of polysaccharides,89,90,93,94 allowing a direct
investigating their effect on the stability, the average comparison with nanospheres obtained from anionic
diameter, and the particle size distribution. However, emulsion polymerization. The first difference is the
it was not fully understood at this time that some of conformation of polysaccharide chains at the surface
them, especially those containing nucleophilic func- of the nanospheres in direct relation with the structure
tional groups, might take part in the initiation of of the copolymer. Indeed, anionic emulsion polymer-
the polymerization, leading to a partial formation ization led to grafted copolymers, whereas linear block

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∗
∗ ∗
O
O O NC OCH3
n
H n OCH3 H n OH
O

(a) (b) (c)

O
O
∗
HO
HO O O
O
OH
O
O
∗ NH2
HO
HO ∗
O
O

COOH
∗
COOH
COOH
COOH

COOH

(d) (e) (f)

FIGURE 7 | Schematic representation of poly(alkyl cyanoacrylate)-based nanospheres with controlled surface properties using poly(ethylene
glycol) monomethyl ether (a), poly(ethylene glycol) (b), poly[(hexadecyl cyanoacrylate)-co-methoxypoly(ethylene glycol) cyanoacrylate] copolymer (c),
polysaccharide chains under anionic initiation (d), polysaccharide chains under redox initiation (e), and amino acids (f). The moiety anchored at the
surface of the nanoparticles is marked by single asterisk.

copolymers were achieved under redox radical ini- is known to play a significant role in the non-
tiation (Figure 8), leading respectively to compact specific recognition events of the immune system.
loops (Figure 7(d)) and hairy polysaccharide chains Indeed, according to Peracchia et al., nanospheres
(Figure 7(e)) at the surface of the nanospheres.88,93 bearing big loops because of α,ω-dihydroxyl PEG
The size of the polysaccharide-decorated (Figure 7(b)) were shown to better prevent comple-
nanospheres was in the 80–800 nm range and ment consumption than do the hairy nanoparticles
depended on: (1) the molecular weight of the polysac- obtained from PEG monomethyl ether87 (Figure 7(a)).
charide, where a minimum value of about 6000 g Besides, Bertholon et al. demonstrated that, for both
mol−1 was required for ensuring an efficient colloidal
dextran and chitosan, an increase of the length of the
stability88,89 and (2) on the nature of the polysac-
compact loops (Figure 7(d)) resulted in an increase
charide: dextran-decorated nanospheres exhibited an
average diameter below 300 nm, dextran sulfate and of complement activation, whereas the opposite effect
chitosan led to a larger average diameter of about was obtained by increasing the length of the hairy
350–600 nm,88 whereas the use of heparin conducted polysaccharide chains88 (Figure 7(e)), which clearly
to 90-nm nanospheres.95,96 demonstrated that complement activation is highly
Another crucial difference resulting from the sensitive to any change of the surface chain conforma-
surface conformation of the hydrophilic chains, for tion. In a recent work, it was also suggested that the
either PEG derivatives or polysaccharides, concerns conformation of the coating material also affects the
the measure of the complement activation,87,88 which cytotoxicity profile of PACA nanoparticles.97

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 WIREs Nanomedicine and Nanobiotechnology Synthesis of PACA-based colloidal nanomedicines

FIGURE 8 | Anionic emulsion polymerization of alkyl cyanoacrylates initiated by hydroxyl groups of dextran (a) and redox radical emulsion
polymerization (RREP) of alkyl cyanoacrylates initiated by dextran/cerium IV (Ce4+ ) ions redox couple (b).

Recently, an interesting synthetic pathway technique which first requires the synthesis of
to functionalize PACA nanospheres using amino amphiphilic copolymers with PEG segments. PIBCA-
acids was proposed by Weiss et al.70 The authors b-PEG diblock and PIBCA-b-PEG-b-PIBCA tri-
used a miniemulsion process to prepare a stable block copolymers were synthesized from phosphine
pH 1 dispersion of nBCA nanoparticles stabilized end-capped PEG macroinitiators.45 With diblock
by SDS as the surfactant. Polymerization was then copolymers, unimodal size distribution and sta-
triggered by the addition of nucleophilic compounds ble nanoparticles in the range of 100–700 nm
such as amino acids (for instance, glycine), leading were obtained by nanoprecipitation or emulsifica-
to functionalized, stable nanospheres (as already tion/solvent evaporation, the average diameter being
discussed earlier, the similar miniemulsion process has controlled mainly by the amount of organic solvent
been applied to nonfunctionalized nanospheres when and by the composition of the polymers. However, the
sodium hydroxide was added as the initiator). This presence of phosphine groups within the synthesized
method allowed: (1) the solids content to be increased polymers may be a toxicological issue.
up to 10 wt% with average diameter ranging from 80 The amphiphilic, biodegradable copolymers
to 350 nm, depending on the amount of surfactant comprising poly(hexadecyl cyanoacrylate) hydropho-
as well as the nature of the amino acid and (2) a bic units and methoxypoly(ethylene glycol) cyano
convenient surface functionalization by amino acid acrylate hydrophilic units (Figure 4) were used to
moieties (Figure 7(f)). prepare the corresponding P(HDCA-co-MePEGCA)
The preparation of ‘PEGylated’ nanoparti- nanospheres exhibiting a biodegradable PACA
cles from preformed polymers is a well-established core and a shell of excretable PEG chains46,98,99

Vo lu me 1, Jan u ary /Febru ary 2009  2008 Jo h n Wiley & So n s, In c. 119

 Advanced Review www.wiley.com/wires/nanomed

PHDCA-co-PEGCA who hypothesized the formation of lipoprotein parti-

0.25 PHDCA cle mimics recognized by the LDLR gene family in the
PHDCA-P80 brain endothelial cells of the BBB.108
PHDCA-Polox 908
0.20
% dose / g tissue

Synthesis of ‘PEGylated’ Nanocapsules

0.15 To the best of our knowledge, the only exam-
ples of ‘PEGylated’ PACA nanocapsules were
0.10 reported by Brigger et al.81 and Li et al.,109,110
both using P(HDCA-co-MePEGCA) copolymers.46
0.05
Although Brigger et al.81 prepared the corresponding
0.00
stealth, oil-containing nanocapsules by the interfacial
a deposition technique, Li et al. used a water-in-oil-in-
b water (w/o/w) double emulsion process to achieve
c
’PEGylated’, water-containing nanocapsules as tumor
FIGURE 9 | Concentration of radioactivity in right hemisphere (a), necrosis factor-α carriers.109,110 This two-step emul-
left hemisphere (b), and cerebellum (c), after intravenous administration sification protocol started by the emulsification of the
of 60 mg kg−1 of [14 C]-P(HDCA-co-MePEGCA) nanoparticles, aqueous phase containing the drug into the organic
poloxamine 908-coated [14 C]-PHDCA nanoparticles, polysorbate phase in which the P(HDCA-co-MePEGCA) copoly-
80-coated [14 C]-PHDCA nanoparticles, and uncoated [14 C]-PHDCA mer was dissolved (w/o), followed by its addition into
nanoparticles (mice at 1 h postinjection). an aqueous PVA solution (w/o/w). Stable nanocap-
sules of about 140–150 nm in diameter were then
(Figure 7(c)). Nanoprecipitation or emulsifica- collected by centrifugation.
tion/solvent evaporation techniques employing
P(HDCA-co-MePEGCA) polymers led to very stable
‘PEGylated’ nanospheres with average diameters Addressed Poly(alkyl cyanoacrylate)
in the 100–200 nm range and monomodal size Biodegradable Nanoparticles
distributions.98 These materials showed a reduced For the forthcoming years, the most exciting chal-
cytotoxicity toward mouse peritoneal macrophages, lenge in drug delivery, irrespective of the nature of the
and the presence of the PEG segments was found drug carriers (i.e., liposome, nanoparticles), will
to increase the degradability of the polymer in the be undoubtedly the synthesis of efficient ligands-
presence of calf serum.98 Besides, as a result of the decorated colloidal devices for achieving specific
PEG coating, an extended circulation time in the cells targeting, on the basis of molecular recog-
bloodstream was demonstrated.100 nition processes. Indeed, the main drawback of
The impressive result deriving from the use previous generation of drug carriers is their non-
of these stealth nanoparticles is their ability to specific drug release behavior. Nanoparticles are
significantly cross the blood–brain barrier (BBB) indeed unable to be efficiently addressed to the
compared with non-PEGylated counterparts and those desired cells and the therapeutic activity of the
with preadsorbed surfactants such as polysorbate 80 encapsulated drug may be partly hampered. Even
or poloxamine 9082,101–104 (Figure 9). for the remarkable case of brain-targeted P(HDCA-
This unique feature suggested that P(HDCA-co- co-MePEGCA) nanospheres,2,101–104 the linkage of a
MePEGCA) nanospheres exhibited appropriate prop- judicious ligand at their surface would certainly result
erties for entering the central nervous system (CNS) in a strongly higher extravasation yield across the
via the BBB. Even though a passive diffusion because BBB.
of an increased permeability of the BBB (when locally Thus, if a great deal of effort has been already
disrupted at the tumor site) may not be ruled out, devoted to this area, a lot of works remain
the mechanism by which those nanoparticles prefer- due to be done. The only example of the so-called
entially crossed the healthy BBB was assigned to a third-generation PACA nanoparticles involves folate-
specific adsorption of apolipoprotein E and B-100 decorated P(HDCA-co-MePEGCA) nanospheres
(Apo E and B-100) onto P(HDCA-co-MePEGCA) to target the folate receptor, which is overex-
nanospheres leading to their translocation mediated pressed at the surface of many tumor cells. For
by low-density lipoprotein receptors (LDLR).105–107 this purpose, the synthetic route for P(HDCA-
The involvement of Apo E on the translocation co-MePEGCA) copolymers46 was adapted to the
through the BBB of polysorbate 80-covered PACA synthesis of a poly[(hexadecyl cyanoacrylate)-
nanoparticles was also reported by Kreuter’s group co-aminopoly(ethylene glycol) cyanoacrylate]

120  2008 Jo h n Wiley & So n s, In c. Vo lu me 1, Jan u ary /Febru ary 2009

 WIREs Nanomedicine and Nanobiotechnology Synthesis of PACA-based colloidal nanomedicines

[P(HDCA-co-H2 NPEGCA)] copolymer, starting (a) CN

CN
from a protected aminopoly(ethylene glycol)
n + n ROH
cyanoacetate.111 O O n
Then, the corresponding nanospheres were R
HO O
obtained by nanoprecipitation showing a narrow
size distribution for an average diameter of 80 nm.
The conjugation with N-hydroxysuccinimide–folate (b) CN CN
(NHS–folate) occurred via an amidation pathway CN
n m + (n – m)
directly at the surface of the nanospheres bearing avail- O O O O O
able amino groups (Figure 10). The specific interaction R R O
occurring between the folate-conjugated nanospheres R
m << n
and the folate-binding protein was demonstrated by
surface plasmon resonance. The apparent affinity of (c) CN
the folate bound to the nanospheres appeared 10-fold CN
higher than the free folate in solution, because of the n n + n CH2O
multivalency of the folate-decorated nanoparticles. O O COOR
R

Biocompatibility and Biodegradation CN

(d)
of Poly(alkyl cyanoacrylate) Polymers HO
CN CN
The degradation and toxicity of PACA nanoparti- n + CH2O
H n –1
O O
cles are a crucial point, especially for biomedical O O O O
R
applications. Indeed, a drug carrier device is suitable R R
for in vivo applications only if it is made of biocom-
patible, possibly biodegradable, or at least excretable FIGURE 11 | Possible degradation pathways for poly(alkyl
cyanoacrylate) (PACA) polymers: hydrolysis of ester functions (a),
(e.g., by the kidneys) materials. In fact, PACAs are
’unzipping’ depolymerization reaction (b), the inverse Knoevenagel
bioerodible polymers for which different degradation
condensation reaction (c), and release of formaldehyde from hydrolysis
pathways have been reported so far (Figure 11). of the α-hydroxyl functions (d).
The predominant mechanism occurs via the
hydrolysis of their side chain ester functions,55,112,113
producing the corresponding alkyl alcohol and
poly(cyanoacrylic acid) as the degradation products, affected by: (1) the length of the alkyl side chains;
the latter being fully water-soluble and readily the longer the alkyl side chains, the lower the toxicity
eliminated by kidney filtration (Figure 11(a)). This but the slower the hydrolysis55,114,115 and (2) the sur-
hydrolysis, which is believed to be the main degrada- rounding environment as it can be strongly catalyzed
tion mechanism in vivo, proceeds typically in a couple by esterases from serum, lysosomes, and pancreatic
of hours for PACA nanoparticles and is strongly juice.116,117 However, a complete excretion of these

∗ O
O

∗ NC n N
H
Folate
O Folate O
NC n NH2
O

O O
Folate Folate
N O

O
Folate

Folate

FIGURE 10 | Synthesis of folate-decorated poly[(hexadecyl cyanoacrylate)-co-aminopoly(ethylene glycol) cyanoacrylate] [P(HDCA-co-H2

NPEGCA)] nanospheres. The moiety anchored at the surface of the nanoparticles is marked by single asterisk.

Vo lu me 1, Jan u ary /Febru ary 2009  2008 Jo h n Wiley & So n s, In c. 121

 Advanced Review www.wiley.com/wires/nanomed

materials would occur only for low-molecular-weight CONCLUSION

PACA polymers, typically below 10,000 g mol−1 .
Even though the chemistry of alkyl cyanoacrylates is
It has been postulated that the ‘unzipping’ not as straightforward as for other ‘common’ vinyl
depolymerization reaction, initiated by a base, could monomers, the convergent involvements of organic
also take part in the biodegradation pathway of chemistry, polymer science, and physicochemistry
PACA,66 especially in biologic media where it can be made possible the development of more and more
theoretically induced by amino acids of proteins (Fig- sophisticated, biodegradable PACA-based nanopar-
ure 11(b)). Following the depolymerization of parent ticles, successfully used as drug delivery devices.
Indeed, from the pharmacology standpoint, PACA
polymers, instant repolymerization to form lower-
nanoparticles fulfill important requirements of ideal
molecular-weight polymers would occur, even if no drug delivery systems: ease and reproducibility of
clear description of this mechanism has been shown preparation, ease of storage and administration in
yet, possibly because of its too fast occurrence to be a sterile form, satisfying drug-loading capacity, low
unambiguously observed. toxicity, excellent biodegradability, and feasibility for
Finally, another suggested mechanism for the scale-up production. By playing with experimental
degradation of PACA polymers is on the basis of conditions (nature and amount of reactants, pro-
cess of preparation, etc.), various types of PACA
the well-known inverse Knoevenagel condensation
nanoparticles can be obtained, each of them exhibit-
reaction, which produces the corresponding alkyl ing specific features regarding the nature of the
cyanoacetate and formaldehyde (Figure 11(c)), even drug and/or the way the drug is encapsulated:
though the release of formaldehyde might also result nanospheres (matrix-type nanoparticles; oil-soluble
from hydrolysis of the α-hydroxyl functions of the drug) or nanocapsules (reservoir-type nanoparticles;
polymer chains, provided the hydroxyl ions have been either oil-soluble or water-soluble drug), nonsurface-
initially used as an initiator69 (Figure 11(d)). How- modified (mainly devoted to MPS organs) or ‘PEGy-
lated’ nanoparticles (long-circulating drug carriers) as
ever, the inverse Knoevenagel condensation reaction
well as ligand-decorated nanospheres (addressed drug
has been reported to a lesser extent in aqueous solu- delivery devices). As a result of constant efforts in
tion at physiological pH and too slow to compete this field, PACA nanotechnologies have thus opened
with the above-mentioned enzyme-catalyzed hydroly- exciting perspectives for the discovery of novel and
sis mechanism.118–120 more efficient nanomedicines.

NOTES
The authors thank Christine Vauthier, François Ganachaud, and Didier Desmaële for their fruitful discussions.
The CNRS and the University Paris-Sud are also acknowledged for financial support.

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