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Cite this: New J. Chem., 2012, 36, 227–240

www.rsc.org/njc PERSPECTIVE
Dendrimer therapeutics: covalent and ionic attachmentsw
Saı̈d El Kazzouli,*a Serge Mignani,a Mosto Bousminaab and
Jean-Pierre Majoral*ac
Published on 08 September 2011. Downloaded by MIT Library on 2/10/2021 12:35:19 AM.

Received (in Montpellier, France) 31st May 2011, Accepted 18th August 2011
DOI: 10.1039/c1nj20459a

This concise review focuses on dendrimers functionalized by different linkers required for covalent
and ionic attachments of drugs to the dendrimer’s surface and highlights the importance of the
chemical nature of linkers for controlled release of free drugs. The stability of linkers under
physiological conditions and their lability under acidic conditions such as those of endosomes and
lysosomes or under enzymatic conditions will be discussed. Especially, we review functionality
of the most recently reported drug–dendrimer conjugates. Then, we give a short comparison of
dendrimer conjugates versus either dendrimer complexes or polymer therapeutics and finally we
briefly summarize the importance of both targeted and nontargeted covalently conjugated drugs.

a
Institute of Nanomaterials & Nanotechnology (INANOTECH), 1. Introduction
Moroccan Advanced Science, Innovation and Research (MAScIR)
Foundation, Avenue de l’Armée Royale, Madinat El Irfane, 10100, During the last decade, pertinent reviews on dendrimers
Rabat, Morocco. E-mail: s.elkazzouli@mascir.com;
Fax: +212 (0)537570880; Tel: +212 (0)537576194 synthesis and application in various areas have been reported,
b
Hassan II academy of Science and Technology, Rabat, Morocco mainly in the field of nanomedicine.1–31 The most attractive
c
Laboratoire de Chimie de Coordination du CNRS, and promising therapeutic applications of dendrimers are in
205 Route de Narbonne, 31077 Toulouse Cedex 4, France. gene delivery and imaging techniques, and as vaccine,
E-mail: majoral@lcc-toulouse.fr
w This article is part of the themed issue Dendrimers II, guest-edited antiviral, antibacterial, anticancer and anti-inflammatory
by Jean-Pierre Majoral. agents. For a recent and very impressive example, in vivo

Saı¨d El Kazzouli was born in Serge Mignani was born in

Beni Mellal (Morocco) in
1975. He obtained his MSc
at the University of Orleans
(France) then in 2004 he
received his PhD in organic
chemistry under the super-
vision of Prof. G. Guillaumet
and Prof. A. Mouaddib from
the University of Orleans. In
2004 he worked at the same
University as a postdoctoral
with Prof. L. Agrofoglio
(nucleoside chemistry) then,
Saı̈d El Kazzouli in 2005 with Prof. S. Berteina-
Raboin in collaboration with
Syngenta firm (Switzerland). In 2006 he joined NCI, NIH in
USA to work as a postdoctoral with Dr V. E. Marquez for
3 years on the design and synthesis of anticancer agents. In 2009
he became a scientist researcher at INANOTECH, Foundation
MAScIR in Rabat, Morocco, where he is working in dendrimer’s
group with Prof. Jean Pierre Majoral. His main research
interests are the design and synthesis of bioactive molecules,
drug delivery as well as catalysis.
France. He joined the Catholic
University of Louvain-la-
Neuve (Belgium) in 1978 for
a PhD training under the
supervision of Prof H. G. Viehe.
He obtained his PhD in 1981.
In 1985–86, he joined the
University of Madison (USA)
as a postdoctoral under the
supervision of Prof B. M.
Trost. In 1981, he joined
Rhône-Poulenc (currently
Sanofi-Aventis) at the Vitry
Serge Mignani Research Center, where he
was a Head of the Medicinal
Chemistry Department and Scientific Director. He is the author
of more than 90 publications and more than 100 patents. He is a
member of the ‘‘Comite´ National de la Chimie’’ (December
2009), of the ‘‘Board of Consulting Editors’’ of Bioorganic &
Medicinal Chemistry Letters and Bioorganic & Medicinal
Chemistry (2003–2008), Vice-President of the ‘‘French-
Japanese Society for Fine and Medicinal Chemistry’’, and
recently he was nominated National Representative from France
to the IUPAC Division of Chemistry and Human Health.

This journal is c The Royal Society of Chemistry and the Centre National de la Recherche Scientifique 2012 New J. Chem., 2012, 36, 227–240 227

studies in mice showed that azabisphosphonate dendrimers
prevent complete ankle joint inflammation effects.32
In the nanomedicine domain, dendrimers are gaining more
and more interest in the area of drug carriers and drug
targeting, because they can combine the required character-
istics of the efficient vectors for specific therapeutic targeting:
(i) surface functionalization with ligands for specific recogni-
tion of receptors or antigens at the desired site, (ii) anchoring
of hydrophilic terminal surface groups to avoid the opsoniza-
tion or the macrophages recognition and elimination by the
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RES (reticular endothelial system),33 (iii) size tuning at the
nanometric scale (smaller than 100 nm) for prolonged
circulating time, (iv) incorporation of probes for in vivo
imaging and monitoring. Dendrimers are therefore powerful
and efficient nanocarriers and their biodistribution and
pharmacokinetic parameters can be controlled with respect
to the number and the nature of termini groups.
Several dendrimer-based nanomedicines have already been
approved by the FDA. The major accomplishment in the
field is the phase I/II clinical trial development of Vivagels
(SPL-7013) by StarPharma (Melbourne, Australia). Vivagels
is an anionic G4-poly(L-lysine)-type dendrimer functionalized
with thirty-two naphthalene disulfonate groups showing a
topical vaginal microbicidal activity. Completed phase I
dose-ranging studies in patients have demonstrated positive
safety, tolerability, and pharmacokinetic properties of
SPL-7013. Mild abdominal pain or discomfort are the main
adverse side effects. These studies represent a bridgehead for
other clinical developments of dendrimers.5
Many reports on drug loaded-dendrimers have shown that
the release of the free drug can be enhanced by suitable choice
of the linker between the dendrimer and the drug, especially,
the linker/spacer length and flexibility. The stability of the
bond between the dendrimer, linker and drug under physio-
logical conditions as well as the lability of the linker–drug
bond in desired tissues are the key-factors for specific drug
Mosto Bousmina was a full
professor and Canada research
Chair on Polymer Physics and
Nanomaterials at Laval
University, Canada, from
1994 to 2008. He then went
to Morocco, where he is
presently the General Director
of INANOTECH and the
Chancellor of the Hassan II
Academy of Science and Tech-
nology. He is the author
of more than 150 scientific
publications and 6 patents.
Mosto Bousmina He received various awards:
Louis-Pasteur Award in
1993, Innovation Award from the Quebec Ministry of
Commerce and Technology in 1998, and Canada-top-20
explorer Award in 2002 from CIAR (Canadian Institute for
Advanced Research), International Morand Lambla Award
from the Polymer Processing Society in 2000 and the prestigious
Steacie Award from NSERC-Canada in 2005.
228 New J. Chem., 2012, 36, 227–240 This journal is
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controlled release. Thus, reaching a proper stability/lability
balance for the linker between the drug and the dendrimer is
critical. This can be achieved by adequate selection of both the
dendrimer end-groups and the linker chemical structure for
each kind of targeted tissue.34 This concise review does not
intend to be exhaustive but aims to report and to discuss the
most recent developments in the field of the dendrimer-based
drug delivery by selecting pertinent and representative
examples. The different kinds of linkers used so far (Fig. 1)
as well as the chemical nature of the attachment of drugs to
dendrimers will be discussed. In addition, we will comment on
the drug release process and biological activities of drug
loaded-dendrimers versus the plain drug whenever they are
studied. We will also review and compare dendrimer
conjugates versus either dendrimer complexes or polymer
conjugates. This review is therefore a literature updated
supplement of the recent reviews reported by Haag et al.,6
Kojima,35 Jain et al.16 and Farokhzard et al.36
2. Covalent drug binding to dendrimer termini
Fig. 1 shows representative examples of drug–dendrimer
conjugate linkers and the corresponding drugs. The main used
points of attachment are: amides, esters, disulfides, hydra-
zones, thiol-maleimide and sulfinyls.
2.1 Amide linkers
One of the earliest studies on drug-loaded dendrimers was
reported by Zhuo et al.37 Poly(amido-amine) dendrimers,
with a 1,4,7,10-tetraazacyclododecane core, were partially
acetylated leading to an enhancement of their water solubility.
5-Fluorouracil (5-Fu), an antimetabolite anticancer drug,
typically administered with leucovorin and acting as an
irreversible inhibitor of thymidylate synthase, was attached
by a simple amide linker to the acetylated dendrimers leading
to 5-FU–dendrimer conjugates (Scheme 1). Assays on the
Jean-Pierre Majoral is an
Emeritus Director of
Researches Exceptional Class
at CNRS. After his PhD he
worked as a Post-doc with
A. Katritzky (Norwich, UK).
He became Directeur de
Recherche at CNRS in 1978.
He was in charge of a French-
Polish European Institute
(LEA, 2000–2008). His
research interest is mainly
focused on main group
elements, especially phosphorus
Jean-Pierre Majoral in different areas of chemistry.
Presently, he is involved in the
preparation and properties of dendrimers and hyperbranched
polymers with emphasis on their applications in different fields
ranging from biology, nanomaterials and catalysis. He is a
member of several Academy of Sciences worldwide and received
various international awards. He is the author of around 500
publications and 40 patents.
c The Royal Society of Chemistry and the Centre National de la Recherche Scientifique 2012
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Fig. 1 Linkers used for the attachment of different drugs onto dendrimers. The arrow indicates the side of site of attachment of the drug to the
dendrimer.
Scheme 1 Poly(amide-amine) dendrimer–5-FU conjugates.
release of the conjugate have shown that hydrolysis in a
phosphate buffer solution (pH 7.4) at 37 1C released free
5-FU. However, the amount of the released 5-FU is influenced
by the conjugates generation and the highest drug release was
obtained with the highest conjugate generation.
Kopecek and collaborators38 have reported the synthesis of
original star-like HPMA copolymers, which were prepared by
the reaction of semitelechelic poly[N-(2-hydroxypropyl)-
methacrylamide] macromolecules (ST-PHPMA), bearing
chemically reactive ester groups, with amino end-groups of
G2, G3, and G4 PAMAM dendrimers. Doxorubicin (DOX),
an anthracycline antibiotic, used in cancer chemotherapy and
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acting by intercalating the DNA, is linked to ST-PHPMA–
DOX (conjugate B) via an amide linkage. The final
ST-PHPMA–DOX dendrimers are obtained with good overall
yields. Additionally, linear N-(2-hydroxypropyl)methacryl-
amide copolymers containing DOX (HPMA–DOX) as well
as the DOX–HPMA copolymer conjugate via a tetrapeptide
sequence (GFLG–DOX, conjugate A) were also prepared. The
cytotoxicity of ST-PHPMA–DOX toward human ovarian
carcinoma A2780 cells was evaluated and compared to the
plain drug (DOX), HPMA–DOX and GFLG–DOX
conjugates. The biological results showed that cytotoxicities
of all conjugates were lower than the free DOX and the most
cytotoxic conjugate was ST-PHPMA–DOX followed by
GFLG–DOX and then HPMA–DOX. The authors proposed
that the differences in cytotoxic activities may result from the
difference in the rate of DOX released by lysosomal enzymes
via amido group cleavage and, probably, the cellular penetra-
tion rate (Scheme 2).
Another example was reported by Jiang’s group39 and
consists of the synthesis of different DOX–PEGylated
PAMAM conjugates. DOX was conjugated to differently
PEGylated PAMAM dendrimers by two different amide link-
age types. The first linker is pH-sensitive (cis-aconityl linkage)
and the second linker is pH-insensitive (succinic linkage),
which leads to PCD and PPSD conjugates, respectively
(Scheme 3). In vitro evaluation studies have shown that
DOX release from pH-sensitive conjugates increased with
increasing PEGylation degree. In addition, cytotoxicity of
pH-sensitive conjugates against ovarian cancer (SKOV-3) cells
increased, while cellular uptake decreased with increasing
PEGylation degree. In contrast, pH-insensitive conjugates
released negligible drug concentration at any tested pH
conditions explaining their low cytotoxicity. The conjugates
were internalized in SKOV-3 cells via successively clathrin-
mediated and adsorptive endocytosis processes. DOX was
New J. Chem., 2012, 36, 227–240 229
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Scheme 2 Structure of HPMA copolymer–DOX conjugates.
Scheme 3 PPCD conjugate and PPSD conjugate.
delivered by acidic lysosomes from pH-sensitive conjugates.
Then, DOX diffused into the nuclei of the cell to induce its
biological activity. Interestingly, pH-sensitive conjugates with
the highest PEGylation degree showed the best tumour accu-
mulation in mice inoculated with SKOV-3 cells.
In another interesting report, Kannan and collaborators40
have studied the cytotoxicity of dendrimers linked with
methotrexate (MTX). MTX is an antimetabolite anticancer
drug acting as an inhibitor of dihydrofolate reductase. The
authors have prepared two different PAMAM dendrimer–
MTX conjugates (Scheme 4) using two different amide linkage
types. The two starting molecules were the anionic half
generation G2.5 PAMAM dendrimer and the cationic G3
PAMAM dendrimer. The first amide bond was achieved by
the reaction between the carboxylic acid-terminated G2.5
dendrimer and the amine group of MTX (conjugate A). The
second amide bond was the result of the reaction between the
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Scheme 4 Synthesis of G2.5-PAMAM–MTX (conjugate A). (for
conjugate B, G3-PAMAM–MTX, a similar procedure was used).
amine-terminated G3 dendrimer and the carboxylic acid group
of MTX (conjugate B). Interestingly, 24-fold improvement of
the half maximal inhibitory concentration value (IC50) was
observed with conjugate A against MTX resistant CEM/MTX
and RII cell lines compared to free MTX. In contrast,
conjugate B showed relatively little cytotoxicity versus
conjugate A. The authors argued that the difference in the
activities between the anionic and cationic conjugates is
probably due to the distinct intracellular release profiles of
the drug from the conjugates.
2.2 Ester linkers
D’Emanuele et al.41 have reported the synthesis and the
in vitro evaluation of new drug carrier systems based on
first-generation G1-PAMAM dendrimers. Terfenadine (Ter)
is an antihistaminic compound formerly used for the treatment
of allergies. Ter is a potent-glycoprotein (P-gp) efflux substrate
and a water-insoluble drug. Ter was attached to the dendrimer
surface using either a succinyl linker (single ester linker)
leading to the G1–suc–Ter conjugate or a succinicdiethylene
glycol linker (double ester linker) leading to the G1–suc-deg–
Ter conjugate (Scheme 5). Two lauroyl chains (L) were
attached to G1–suc-deg–Ter and give the L2–G1–suc-deg–Ter
derivative which led to an enhancement of its permeability in
the Caco-2 model (the Caco-2 monolayers model for the
prediction of intestinal drug absorption). All conjugates were
more hydrophilic than the free drug and the biological assays
indicated that the conjugates had no impact on the viability of
Caco-2 cells up to a concentration of 1 mM.
Baker, Jr et al.42 have synthesized PAMAM dendrimer-
based multifunctional therapeutic devices. In addition to
MTX which was attached to the PAMAM dendrimers via
an ester linker, imaging agents such as fluorescein isocyanate
(FI) or fluorescein isothiocyanate (FITC) were attached
through urea or thiourea linkage. Moreover, folic acid (FA)
was attached via an amide function and was used as a specific
targeting agent (Scheme 6). In other reports, Baker, Jr and
co-workers43,44 have shown that the G5–FI–FA–MTX conju-
gate binds, internalizes, and induces cytotoxicities specifically
in the overexpressed FA receptor-expressing KB cells in vitro.
This interesting result demonstrates clearly the applicability
of dendrimers for the specific delivery of drugs into cells.
c The Royal Society of Chemistry and the Centre National de la Recherche Scientifique 2012
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Scheme 5 G1–suc-deg–Ter and L2-G1–suc-deg–Ter conjugates.
Scheme 6 Schematic representation of the G5–FITC–FA–MTX
conjugate.
Recently, G3–PAMAM–FI–MTX was synthesized by linking
both MTX and FI via ester bonds. The conjugate was able to
target FR-expressing cells without the need to conjugate FA.45
In a recent report, Haandel and Stobaugh46 have developed
a bioanalytical assay for the detection of the G5–MTX–FA
conjugate, and the released MTX as well as its main plasma
metabolite 7-hydroxymethotrexate. Recently, Baker’s group47
has published an interesting work on the synthesis of
G5–MTX conjugates via an ester linkage. They used riboflavin
(RF) as a specific targeting ligand (Scheme 7). Thus, the
G5–FITC–RF–MTX conjugate was synthesized and was
evaluated in vitro against KB cells. The biological results have
shown that the conjugate was carried into the cells through a
RF-receptor mediated mechanism, which can lead to a specific
release of MTX.
Following the same approach, Baker’s group48 has
synthesized the G5–FI–FA–paclitaxel conjugate. Paclitaxel
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Scheme 7 G5-PAMAM–RFn–MTXm conjugate.
(also known as Taxols) is a well known mitotic inhibitor used
in cancer chemotherapy. The synthesis started by the neutra-
lization of primary amino groups on the surface of the
G5-PAMAM dendrimer by partial acetylation, leading to
enhanced solubility of the dendrimer itself. Then, FITC, FA
and paclitaxel were successively conjugated to the remaining
nonacetylated primary amino groups using thiourea, amide
and diester linkages, respectively (Scheme 8). These multi-
functional dendrimer conjugates have been tested in vitro for
targeted delivery of chemotherapeutic and imaging agents to
specific cancer cells. In vitro testing has shown uptake and the
cytotoxic effect of the conjugate.
In another interesting report, the kinetic rate of the hydro-
lysis of the dendrimer-Taxol conjugates in phosphate buffered
saline (PBS) was investigated.49 For the preparation of
G5–FITC–FA–Taxol conjugates, the Taxol moiety was linked
to the G5-PAMAM dendrimer unit using either succinic acid
(G5-PAMAM–FITC–su–Taxol) or glutaric acid (G5-PAMAM–
FITC–glu–Taxol) linkers (Scheme 9). This study showed that
the succinic acid linker–dendrimer conjugate released the
Taxol drug through bond hydrolysis in a time-dependent
fashion with a half-life of 10 hours. In contrast, the glutaric
acid linker–dendrimer conjugate was very stable and did not
release the Taxol compound even after seven days under
similar experimental conditions. Strongly, this result confirms
the use of succinic acid as a useful linker.
Chai and collaborators50 have reported an elegant synthesis
of novel water-soluble and photostable dendrimers containing
the 3,4-dihydroxy-L-phenylalanine (L-DOPA) scaffold, which
Scheme 8 Schematic representation of the G5–FITC–FA–Taxol
conjugate.
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Scheme 9 G5-PAMAM–FITC–su–Taxol and G5-PAMAM–FITC–

su–Taxol conjugates.

is a commonly used drug to treat Parkinson’s disease. The

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L-DOPA-dendrimers were synthesized by connection of indi-

vidual L-DOPA moieties to one another via hydrolysable
diester linkages (Scheme 10). A G3-generation L-DOPA
dendrimer, containing 30 L-DOPA residues, showed a 20-fold
increase in aqueous solubility and enhanced photostability in
solutions over L-DOPA under identical experimental condi-
tions. L-DOPA-dendrimers represent a useful biocompatible
dendrimeric scaffold similar to the well known examples of
polyether-scaffold dendrimers,51 polyester-scaffold dendrimers52
and recently polypropylimine-scaffold dendrimers.53
Lim and Simanek54 have described the synthesis of triazine
dendrimers bearing the Taxol moiety on their surfaces (Scheme 11).
Scheme 11 Schematic representation of the Taxol loaded triazine
dendrimer.
These new dendrimer conjugates contain two Bolton–Hunter-
type groups, used for biodistribution studies, and multiple
Taxol groups attached by labile ester linkage. The conjugates
were PEGylated leading to a water soluble drug–dendrimer
conjugate.
In another study, Kannan et al.55 have reported the synthesis
of simple hyperbranched polyol scaffolds and G4-OH PAMAM
dendrimers bearing methyl prednisolone (MP) (Scheme 12).

Scheme 10 Structure of the G3-L-DOPA dendrimer.

232 New J. Chem., 2012, 36, 227–240 This journal is c The Royal Society of Chemistry and the Centre National de la Recherche Scientifique 2012
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Scheme 12 MP–G4-PAMAM dendrimer conjugate.
MP is an adrenocortical steroid typically used in the anti-
inflammatory field. In this study, the MP was conjugated to
PAMAM dendrimers and polyol scaffolds using either a
glutaric acid (GA) or a succinic acid (SA) spacer. The number
of drug payload was the highest with polyol whatever the
spacer type, while in the case of the PAMAM dendrimer, the
higher payload was achieved with the GA than the SA spacer.
The anti-inflammatory activity of these MP–polymer conju-
gates was assessed by the evaluation of the inhibition of
prostaglandin synthesis. The inhibition potency follows the
order: MP–SA–dendrimer conjugate 4 MP–GA–polyol con-
jugate 4 MP–GA–dendrimer conjugate. In conclusion, in this
study, Kannan’s group has shown that drug release from
hyperbranched polymer–drug conjugates and the subsequent
activity are influenced by the branching architecture and the
type of the linker. Interestingly, the same group has conju-
gated MP to the G4-PAMAM dendrimer using a diester
linkage.56 The intranasal administration of the MP–dendrimer
conjugate was effective in reducing the ovalbumin-induced
airway inflammatory response in a mouse model. Moreover,
significant efficacy of the drug was observed even at the lowest
dose tested and the MP–dendrimer conjugate was able to
reduce the inflammatory reaction. It is noteworthy that the
MP–dendrimer conjugate has no undesired effects such as
non-specific inflammation and has a longer residence time in
the lungs.
In continuation of their work, Baker, Jr et al.57 have
reported the simultaneous conjugation of FA and MTX to
the G5-PAMAM dendrimer through ester linkages in a ‘‘one
pot’’ approach (see Scheme 4). In vitro studies performed on
FA receptor-expressing KB cells have shown that the
G5–FA–MTX conjugate has a similar affinity and cytotoxic
potency to G5–FA–MTX synthesized using the traditional
multiple-step approach.
2.3 Disulfide linkers
Kannan et al.58 have reported the synthesis of PAMAM-
COOH containing N-acetyl cysteine (NAC, Fluimicils). NAC
was used in different therapeutical indications such as
anti-inflammatory and for the treatment of paracetamol
(acetaminophen) overdose. The conjugate was prepared using
a disulfide linker (glutathione sensitive linker) which enables
relatively rapid intracellular release of the free drug.
The new conjugate is non-toxic even at the highest concentra-
tion tested in vitro and is a more effective antioxidant and
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Scheme 13 Cationic G4-NH2 and anionic G3.5-COOH PAMAM
dendrimers bearing NAC.
anti-inflammatory agent, when compared to free NAC in vitro.
Even at a low dose (0.5 mM of NAC) the NAC–dendrimer
conjugate showed a high inhibitory rate. The same group59 has
reported the synthesis of two different NAC–G4-PAMAM
dendrimer conjugates using two disulfide linkers (GSH-sensitive
linker): the first conjugate with a cationic G4-PAMAM–NH2
and the second with anionic G3.5-PAMAM–COOH dendri-
mers (Scheme 13).
The results of NAC release from the conjugates have shown
that 70% of NAC payload was released at intracellular GSH
concentration and only negligible NAC release was observed
at extracellular GSH levels. FITC-labeled conjugates showed
rapid penetration and localization in the cytoplasm of lipo-
polysaccharide (LPS)-activated microglial cells. Both G4-NH2
and anionic G3.5-COOH PAMAM–NAC conjugates showed
better nitric inhibition when compared to free NAC.
2.4 Hydrazone linkers
To date, few works on dendrimers bearing drugs
through hydrazone bonds have been reported. Fréchet and
collaborators60 have synthesized dendritic polyester systems
using 2,2-bis(hydroxymethyl)propanoic acid as the building
block. This new family of dendrimers has excellent water
solubility. To investigate the feasibility of the drug–dendrimer
conjugate, authors have successfully attached DOX using a
hydrazone bond (pH sensitive linkage) (Scheme 14).
Recently, the same group61 has reported the synthesis of a
new tubulysin analogue, a potent anticancer drug, which was
also conjugated via a hydrazone linker (acid labile bond) to the
PEGylated dendrimer with a polypeptide core. The conjugate
has a higher water solubility compared to the free tubulysin
analogue and shows no sign of toxicity, even at high doses of
the conjugate. In addition, the conjugate exhibited a longer
lifespan (90% increase on average) (Scheme 15).
5-Aminosalicylic acid (5-ASA), an anti-inflammatory agent,
was attached to the PAMAM dendrimer using two different
spacers, p-aminobenzoic acid (PABA) and p-aminohippuric
acid (PAH). The new water-soluble PAMAM dendrimer–
5-ASA conjugates with a designed spacer for colonic delivery62
have shown colon-specific and gradual release of 5-ASA after
incubation with rat cecal contents at 37 1C (45.6% from
PABA linker/dendrimer and 57% from PAH linker/dendrimer
conjugates). Based on these results Wiwattanapatapee et al.62
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Scheme 14 Dendritic polyester–DOX conjugate.
Scheme 15 Tubulysin analogue–PEGylated dendrimer conjugate.
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Scheme 16 Glycopeptide dendrimer–colchicine conjugates.
have suggested that these dendrimers have potential for use as
colon-specific drug carriers.
2.5 Thiol-maleimide and sulfinyl linkages
Barth and coworkers63,64 have reported the first example of
urea linkage using thiol-maleimide coupling for the synthesis
of boronated monoclonal antibodies–dendrimer conjugates.
These conjugates were designed as delivery systems for
neutron capture therapy. Later, Wängler and collaborators65
have loaded DOTA onto the PAMAM dendrimer using the
same linker type.
Colchicine, which is a natural product with high cyto-
toxicity, was loaded onto the peptide dendrimer using sulfinyl
acetamide66 (Scheme 16). In contrast to free colchicine,
biological results have shown that colchicine–dendrimer
conjugates present selective cytotoxicities toward cancer cell
lines, such as HeLa cells, than normal cells.
3. Ionic attachment to dendrimer termini
(cisplatin and copper)
In drug delivery approaches by dendrimers developed so far,
cisplatin (trade names Platinol and Platinol-AQ) is the most
investigated metallodrug. Cisplatin is a potent anticancer
agent used in chemotherapy to treat various types of cancers.67
Thus, cisplatin was loaded into dendrimers using two
different coordination strategies (Scheme 17). (i) (NH2)2-Pt–
(dendrimer)2: the bonds between the platinum entity and the
dendrimer should be labile and should lead to the release of
the free drug, ‘Cl2Pt(NH2)2’.68 (ii) (Cl)2-Pt–(N-dendrimer)2:
the amine group on the dendrimer (at least one N–H moiety)
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Scheme 17 Synthesis of cisplatin dendrimer conjugates.
could establish hydrogen bonding and the conjugates may bind
directly to DNA without the release of the free cisplatin.69
In early studies, Malik et al.70 conjugated G3.5 PAMAM-
CO2H dendrimers with cisplatin giving PAMAM–platinate
complexes (mono-, bi- and cross-linked-dentates). These
complexes are less toxic than cisplatin itself and release
platinum slowly in vitro. In in vivo tests, the dendrimer–Pt
displayed similar potency as cisplatin administered by the
intraperitoneal route against the L1210 leukemia tumour
model in mice. At high doses, dendrimer–Pt, given by the
intraperitoneal route, showed good activity against the
B16F10 melanoma model whereas cisplatin did not. Intravenous
administration of dendrimer–Pt induced potency against the
subcutaneous B16F10 model, while cisplatin was inactive. After
intravenous injection of dendrimer–Pt and cisplatin, selective
accumulation of dendrimer–Pt in solid tumour versus blood
was observed probably due to the EPR effect (vide supra).
Biocompatible anionic linear-globular dendrimer–cis–
platinum(II) conjugates have been described.71 The G2-citric-
acid-based dendrimer showed potent in vitro cytotoxic activities
against HT1080 and CT26 cell lines (up to 8 fold versus plain
cisplatin) and, interestingly, against the Pt resistant cell line,
SKOV3 (B2 fold versus plain cisplatin), whereas the corres-
ponding G1 dendrimer showed lower potencies. Hemolytic
impacts and cell death mechanisms of these conjugates on
human blood and HT1080 cell line were investigated as well.
The same hemolysis behaviour (1.5, 3 and 6 mg ml1) was
observed for all conjugates except for the G2–Pt complex at
the concentration of 6 mg ml1. Both apoptosis and necrosis
mechanisms (B2 fold versus cisplatin) were attributed to
conjugates and cisplatin in a direct correlation between the
concentration and the degree of cell death.
Kokotos and coworkers72 have synthesized poly-propylene-
imines containing chiral amines with 8, 16 and 32 amino
groups on the surface. Dendrimers with cisplatin moieties at
the periphery were obtained by the reaction between the
free amine dendrimers and potassium tetrachloroplatinate(II)
leading to highly insoluble conjugates.
Chu and his group73 have recently prepared platinum and
copper metallodendrimer conjugates (Scheme 18).
The condensation reaction of platinum chloride and copper
chloride afforded G0-PAMAM–py4–[PtCl2]4 and G0-PAMAM–
py4–[CuCl2]7, respectively. The conjugate cytotoxicities were
studied toward leukemia cells, breast cancer cells and Chang
Liver cells. G0-PAMAM–py4–[PtCl2]4 was inactive to MCF-7
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Scheme 18 G0-PAMAM–py4–[PtCl2]4 and G0-PAMAM–py4–[CuCl2]7
conjugates.
and Chang liver and slightly active toward MOLT-4 cells.
However, G0-PAMAM–py4–[CuCl2]7 displayed higher cyto-
toxicity than both platinum conjugates and free cisplatin itself
against MOLT-4 and MCF-7 cells. This difference in cyto-
toxicity may be attributed to the difference in solubility,
number of metals loaded onto the dendrimer and their
coordination type with the ligands. Other types of conjugates
have also being reported using ionic attachments. For
example, cis-dichlorodiammineplatinum(II), a derivative of
cisplatin, was loaded onto dendrimer phthalocyanine and
poly(ethylene glycol)-block-poly(L-aspartic acid)74 which led
to polymer–metal complex micelles.75
4. Comparison between different linkers
Various reports have investigated the difference between
linkers and their effect for the release of drugs under similar
experimental conditions. In this paragraph, we review the
different studies reported so far.
Baker et al.76 have reported the synthesis of the trifunctional
G5-(Ac)96–FITC5–FA3–MTX4 conjugate which provide a
targeting, imaging, and intracellular delivering system.
MTX was conjugated either through an amide linkage (non
biodegradable) or through an ester linkage (hydrolyzed in the
endosome at low pH). Although both types of MTX triple
conjugates were internalized into KB cells in vitro, only the
MTX conjugated through an ester linkage was cytotoxic. The
conjugates (G5–FITC–FA) with amine-surfaced nanodevices
showed minimal uptake at 30 nM with KB cells and G5–FITC
gave similar uptake, which proved that the binding of both
conjugates (G5–FITC–FA and G5–FITC) was not specific to
the FA receptor. However, the modified surfaces containing
either carboxy, 2,3-dihydroxy propyl or acetamide moieties on
the nanodevice surfaces bind efficiently and rapidly to KB
cells. In this case, the controlled nanodevice without FA did
not bind to KB cells. It is noteworthy that an equimolar
concentration of the free drug (MTX) was 4-fold less effective
in killing the tumour cells than the drug conjugated through
ester linkage. The authors observed also that a drug conju-
gated through amide linkage, and the conjugate without a
drug were not toxic at any tested concentration.
Using polyester dendrimer–PEO hybrids, Fréchet and
collaborators77 have reported the synthesis of DOX–[G3]-
(PEO5k)8-[G4]-(OH)16 bow-tie dendrimers using either
carbamate (stable) or acyl hydrazone (pH-labile) linkages.
New J. Chem., 2012, 36, 227–240 235
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Scheme 19 G0-PAMAM–Naproxen conjugates.
The DOX–dendrimer conjugate through hydrazone linkage
was eliminated from serum with a half life of 16 hours and
its tumour uptake was nine-fold higher than free DOX at
48 hours. It was noticed that a single injection of the
DOX-dendrimer (20 mg kg1 DOX equivalents), 8 days after
tumour implantation, caused complete tumour regression and
100% survival of the mice over the 60-days experiment.
Biological results have shown also that no cure was achieved
in tumour-implanted mice treated with free DOX at its
tolerated dose, a drug free dendrimer, and DOX–dendrimer
through carbamate linkage.
D’Emanuele et al.78 have reported the synthesis and evalua-
tion of G0-PAMAM-Naproxen conjugates, which were
designed in order to enhance drug solubility and bioavailability.
Naproxen, a poorly water-soluble anti-inflammatory drug,
was directly conjugated to the G0-PAMAM dendrimer via
an amide bond or via an ester bon7d using either l-lactic acid
or diethylene glycol as a linker (Scheme 19). The synthesized
conjugates have shown better water solubility than free
Naproxen. All conjugates were chemically stable over 48 h
of incubation at different pH values such as: hydrochloric acid
buffer (pH 1.2), phosphate buffer (pH 7.4), and borate buffer
(pH 8.5). However, Naproxen was enzymatically released
from both ester conjugates in plasma (80% human plasma)
and the release from the lactic ester conjugate was slower than
from the diethylene glycol ester conjugate.
As part of their research program, Kono and coworkers79
have synthesized G4-PAMAM dendrimers having a glutamic
acid (Glu) residue as the end group, then, they have attached
PEG chains to amino groups of Glu residues. DOX was
conjugated to side chains of the Glu residues using either an
amide bond, [PEGeGlu(DOX)-G4], or a hydrazone bond,
[PEGe Glu(NHNe DOX)-G4]. Biological results have shown
that only dendrimers bearing DOX through hydrazone
linkage were stable under physiological conditions, and
released free DOX drug under acidic conditions such as
those of endosomes and lysosomes. These new conjugates
showed much lower toxicity against HeLa cells than free
DOX. In addition, PEGeGlu(NHNeDOX)-G4 was 7-fold
cytotoxic than PEGeGlu(DOX)-G4, which demonstrates
the importance of pH-sensitive hydrazone linkage for high
cytotoxicity.
236 New J. Chem., 2012, 36, 227–240 This journal is
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Scheme 20 Dendrimers 1–3 with 2 0 -ester and disulfide linkages.
In another report, Simanek et al.80 have reported the
synthesis of triazine dendrimers bearing Taxol using either
ester or ester/disulfide linkages (Scheme 20). Two different
PEG-N-hydroxysuccinimide (PEG-NHS) esters were used: for
conjugates 1 and 2, the PEG incorporates an ester-linked
succinic acid group, whereas, the PEG attached to conju-
gate 3 incorporates an ether-linked acetic acid group
(Scheme 20).
In this study, it was shown that the three conjugates
(1, 2 and 3) appear to be well tolerated in vivo at doses equal
to 200% the maximum tolerated dose of Taxols.
In addition, the tumour localization of the conjugates was
observed at low levels and the elimination half-lives of the
three conjugates were similar.
Recently, Kannan and coworkers81 have published a very
interesting work comparing the drug release from polymer–
drug conjugates using different linkers. To understand the
architecture and the linker effect on the release of ibuprofen, a
non-steroidal anti-inflammatory drug, they synthesized
G4-PAMAM dendrimer–ibuprofen conjugates using ester,
amide, and peptide linkers, in addition to a linear mPEG–
ibuprofen conjugate. Comparing the amide linker versus the
ester linker, the amide was more stable against hydrolysis,
whereas the ester-linked conjugates showed pH-dependent
release. This study has shown also that direct linked conju-
gates (amide- and ester) did not release ibuprofen enzymati-
cally in cathepsin B buffer and diluted human plasma. In
contrast, the mPEG conjugate released 65% of its payload
within 12 h in diluted plasma by esterase activity, and the
peptide-linked dendrimer conjugate released 40% of its
payload within 48 h by cathepsin B activity. It is noteworthy
that the steric crowding at the surface of PAMAM
dendrimer–drug conjugates, along with linking chemistry
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Scheme 21 Surface modification using different amino acid moieties.
govern the drug release mechanism as well as its kinetics.
Using amino acids as linkers, Kannan’s research group82 has
prepared new biocompatible dendrimer scaffolds which could
be used in drug delivery. The peripheries of G4-PAMAM
dendrimers were converted in one-step into diverse peripheral
groups (Scheme 21) which could be chemoselectively conju-
gated to different drugs. Dexamethasone and indomethacin,
which are widely used anti-inflammatory drugs, were conju-
gated to the functional dendrimers using ester and amide
linkages, respectively, while the imaging agent FITC was
attached through a thiourea linkage type. The in vitro
cytotoxicity and hemolysis assays have shown that the hetero-
bifunctional dendrimers were noncytotoxic even at high
concentrations (100 mg mL1 to 1 mg mL1).
Good and collaborators83 have shown that sialic acid
functionalized dendritic polymers can act as mimics of cell
surface sialic acid clusters and attenuate Ab-induced neuro-
toxicity. In continuation of their efforts in the development
of treatments for Alzheimer’s disease, Good et al.83 have
developed sialic acid dendrimer conjugates. Thus, two
different attachments were used to link sialic acid to PAMAM
dendrimers. The first linker was the result of the coupling
between PAMAM termini and the anomeric hydroxyl group
of sialic acid (physiological attachment). The second bond
was obtained by the reaction between the PAMAM
termini and carboxylic acid (nonphysiological attachment)
(Scheme 22). Biological results have shown that the use of
the attachment of sialic acid via the anomeric hydroxyl
instead of attachment via the carboxylic acid did not give a
significant enhancement of their ability to mitigate Ab induced
neurotoxicity.84
Scheme 22 Sialic acid PAMAM conjugates using two different
linkages (conjugate A: carboxylic acid, conjugate B: anomeric
hydroxyl group).
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5. Dendrimer conjugates versus dendrimer
complexes or polymer conjugates
5.1 Conjugates versus complexes?
In vitro and in vivo comparative studies of non-covalent drug
inclusion complexes and covalently conjugated drugs have
been carried out by different teams.
Briefly, one of the major potential advantages of covalent
attachment of drugs to the termini surface groups of
dendrimers in comparison with the encapsulation technique
is the better control over drug release. In fact, Baker and
co-workers25 have studied the release kinetics of methotrexate
from the folate–G5-PAMAM-dendrimer–methotrexate conjugate
and folate–G5-PAMAM-dendrimer–methotrexate encapsula-
tion. The authors emphasized that the dendrimer–drug
conjugate was more suitable for targeted drug delivery, while
the complex improved the solubility of methotrexate in
aqueous solution and displayed similar cytotoxicity against
KB cancer cells than plain methotrexate.
In addition, in a recent report, Jiang and coworkers85
presented the comparative evaluation of in vitro and in vivo
behaviors of methotrexate coupled to PEGylated or
non-PEGylated G4-PAMAM dendrimers both encapsulated
and conjugated. The in vitro studies have shown that there
were no strong differences between complexes and free
methotrexate in cytotoxicity against KB cell lines, whereas
the conjugates showed lower potency. Remarkably, in vivo
studies in rodents have shown that the methotrexate–
G4-PAMAM-dendrimer–PEG-conjugate exhibited a good
prolonged blood residence time (in normal rats, i.v. adminis-
tration) and strongest antitumour activity (in S-180 tumour-
bearing mice, i.v. administration).
The need to conduct more systematic investigations for
drugs complexed or conjugated with dendrimers on the long-
term pharmacodynamic (PD) and pharmacokinetic (PK)
behaviors has been brought up by Cheng and Xu.86
5.2 Polymer therapeutics or dendrimer therapeutics?
As shown in Table 1, Patel et al.87 have nicely summarized the
main properties of dendrimers versus linear polymers. Briefly,
many polymers are highly heterogeneous whereas size and
molecular mass of dendrimers can be highly controlled. For
these reasons, dendrimers display significant improvements
when compared to linear polymers related to their physical
characteristics such as polydispersity, structure, structural
control, architecture and compressibility as well as their
chemical characteristics such as easy synthesis and aqueous
solubility. The architecture of dendrimers induces a large
proportion of group exposition at their surfaces affording a
very high molecular surface to volume ratio (41000 m2 g1) in
contrast to classical linear polymers (80–300 m2 g1).
The main biological applications of polymer therapeutics
(biopolymers) encompass polymer–protein conjugates and
drug–polymer conjugates.88 Numerous polymer–protein
conjugates have been developed in order to improve the
stability and the pharmacokinetic behaviors (prolongation of
the biological half-life) and to reduce the immunogenicity of
considered native proteins. PEG-polymers have mainly been
New J. Chem., 2012, 36, 227–240 237

Table 1 Main properties of dendrimers and linear polymers (adapted from Patel et al.)85
Properties Dendrimers
Programmed release of drugs High ( ) reduced toxicity, increased bioavailability, simplified dosing schedule )
Penetration abilities High (high cell membrane penetration ) an increased cellular uptake level of
the drugs complexed or conjugated)
Penetration and retention Enhanced ( ) preferential uptake of the dendrimers by cancer tissues)
(EPR) effect
Immunogenicity Low
a
Size, shape, surface properties of the polymeric carriers highly influence the pharmacodynamic (PD) and pharmacokinetic (PK) behaviors of
drugs encapsulated in/complexed to/conjugated to the dendrimers.
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the polymers developed for the construction of polymer–
protein conjugates. For instance, PEGylated L-asparaginase,
adenosine deaminase, interferon-a (INF-a), styrene-co-maleic
anhydride-neocarzinostatin (SMANCS) and granulocyte
colony stimulating factor (PEG-G-CSF) were described.89
Interestingly, the synthesis of the PEGylated 125I-EGF-OX26
monoclonal antibody construct has been described for
imaging human brain tumours.90
Development of drug–polymer conjugates with cleavable
linkers to improve the therapeutic index of several toxic drugs,
especially in the cancer chemotherapy, has been described.
Currently, phase I and phase II clinical trials are ongoing or
completed with different drugs such as, doxorubicin, taxol,
camptothecin, methotrexate and platinate(II) derivatives. The
main copolymers used are HPMA (N-(2-hydroxypropyl)-
methacrylamide), PEG, poly-L-glutamate, albumin, dextran
and 6-maleimidocaproyl hydrazone derivatives. Different
linker types were employed, namely: Gly-Phe-Leu-Gly,
Gly-6-aminohexanoyl-Gly, alanine ester, Gly-ester, ester and
acid-sensitive hydrazone.88,89
In addition, self-assembling block copolymer micelles have
also been explored as drug carriers. For instance, pluronic
block copolymer micelles including doxorubicin are able to
circumvent p-glycoprotein-mediated resistance which has
shown promising results in phase II clinical trials. The drug
was noncovalently entrapped in the micelle.91 A dual doxo-
rubicin covalently bonded to the PEG-poly(aspartic acid)
copolymer entrapping also free doxorubicin has been
described by Kataoka and colleagues.92 This doxorubicin-
copolymer accumulates preferentially in tumour tissue
through the EPR effect inducing its biological anti-cancer
activity.
One other important biomedical application of biopolymers
is the destruction of angiogenic vasculature of solid tumours.
Several targeted delivery systems were described such as:
polymer–angiogenesis inhibitor conjugates (e.g. HPMA-
TNF-470), antiendothelial immunoconjugates (e.g. L19-inter-
leukin 2, L19-astatine 211), antiendothelial fusion proteins
(e.g. endostatin-human prolactin antagonist), antiendothelial
peptide conjugates (e.g. NGRpeptide–doxorubicin) and anti-
endothelial liposomes (e.g. APRPGpeptide–liposome–
DPP–CNDAC (antitumour nucleoside)). These approaches
showed very promising results in in vitro and in vivo models
and had apparently no specific toxicities. Combination of
polymeric therapeutics in cancer chemotherapy including
oxidative stress inducers or antiangiogenic agents has been
also designed.88,89
238 New J. Chem., 2012, 36, 227–240 This journal is
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Linear polymers
a
High/low
No EPR effect
6. Targeted or nontargeted covalently conjugated
drugs?
In drug delivery by dendrimers, two different approaches have
been developed: active targeted (few examples) and passive
non-targeted drug–dendrimer conjugates (numerous examples).
For example, in the oncology domain, both targeted and non-
targeted drug–dendrimer conjugates can successfully extra-
vasate across tumour’s leaky vasculature and accumulate in
the cancer tissue.
Briefly, targeted drug–dendrimer conjugates have the
advantage to selectively bind cancerous cells versus normal
healthy cells. The receptor on the surface of the cancer cells
increases the residence time on the cell surface and enhances
the internalization kinetics into cells.14 In addition, targeted
drug–dendrimer conjugates allow the release of higher drug
dose loaded in the ‘‘right place’’ (not a premature release of the
drug under biological conditions). Interestingly, Baker and
co-workers have carried out outstanding kinetic studies
showing that the targeted dendrimer conjugates are preferen-
tially taken up by the targeted cancer cells resulting mainly due
to their longer residence times versus the endocytic rate.93
7. Concluding remarks and perspectives
The chemical nature of the linker between the drug and the
dendrimer has a pivotal role in the design of efficient drug
delivery systems. This review summarizes the recent develop-
ments in the design of covalent linkers such as amides, esters,
diesters, disulfides, hydrazones, thiol-maleimide and sulfinyls
and their efficiency in terms of drug delivery and therapeutic
biological activity. The cases of the ionic attachments for the
metallodrugs cisplatin and copper chloride were reported
and discussed as well. Some of the reviewed linkers are
pH-sensitive and have proven to enhance intracellular release
of the free drug. The biological evaluations have shown that,
in most cases, the conjugation of a drug with a dendrimer via a
linker leads to great enhancement of the biological activity
along with the reduction in toxicity compared to free drugs.
For example, in the case of nanodevices engineered for cancer
therapy, the pH-sensitive linkers can be cleaved more rapidly
in cancer cells (endo-lysosomes) than under normal physio-
logical conditions (normal cells). However, the nonspecific
release of free drugs both in cells and tissues remains a major
challenge. To overcome the difficulty, new delivery systems
containing the drug and targeting ligand, both connected to
the same dendrimer (preferentially PEGylated to guarantee
c The Royal Society of Chemistry and the Centre National de la Recherche Scientifique 2012

biocompatibility and long blood circulating time with a
sustained release effect), are under active investigation.
In addition, the multi-functional nature of the dendrimer
surface allows combining drugs, solubilising groups such as
PEG chains in the same surface unit. One of the important
additional advantages of the introduction of a PEG chain is
the inhibition of the blood serum opsonins by the PEGylated
dendrimer allowing its non-recognition by the phagocytic
cells. It is important to note that the surface-modified systems
can operate differently than the native dendrimers. The type of
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linker used to covalently link drugs to the surface termini of
the dendrimers affects directly the biological activity of the
drug–dendrimer conjugates and can be used to control the rate
of drug release (vide supra). The nanometric size of dendrimers
induces their entry into hyper-permeable vasculature of
tumour or inflamed tissues by the enhanced permeability
and retention effect (EPR effect), whereas their high molecular
weight and the lymphatic dysfunctioning causes their specific
localization. In addition, the presence of hydrophobic terminal
surface groups on the dendrimers prevent their recognition by
macrophages and their elimination by the Reticuloendothelial
System (RES).14,94,95
Moreover, the development of new drug-loaded dendrimers
cleavable by specific enzymes and other biological molecules
present exclusively in the targeted cells is also a very attractive
research area and may resolve the problem of non-specific
release. These novel approaches open new avenues to cancer
and other disease treatments. It’s not only a supplement to the
conventional chemotherapy and radiotherapy but also prevents
damage of normal tissues and prevents drug resistance.
We expect that future research on the development of drug-
linked dendrimers will face different important remaining
challenges which could be summarized as follows:
(a) Increase the degree of control in drug delivery by the
design and the synthesis of novel specific linkers that will be
recognized and selectively cleaved by biological molecules
present exclusively in the desired targeted cells versus normal
cells. The development of cell-based therapies for diseased
tissues is probably one of the most promising research fields in
regenerative medicine.
(b) Synthesis of dendrimer engineering towards tissues or
specific cellular compartments targeting (e.g. lung, skin,
nucleus, mitochondria . . .).
(c) Development of clusters of dendrimers for the assembly
of multifunctional therapeutic systems (e.g. combined anti-
cancer therapy . . .).
(d) Synthesis of biocompatible and biodegradable dendri-
mers based on simplified and optimized synthetic pathways
reducing cost of their production.
(e) Set-up appropriate in vivo studies to evaluate the
biocompatibility and the therapeutic profiles of drug–dendrimer
conjugates (PK/PD behaviors).
(f) Ensure the preparation of dendrimers in large quantities
at clinical-grade purity for clinical trials (under GMP conditions).
(g) Implementation of dendrimers technology for the
deployment of personalized medicine via the development of
combined companion diagnostics/therapeutics.
All these aspects should bring about increase of number of
commercialized dendrimers based drug delivery systems
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focused on specific disease at the cellular level (e.g. cell mem-
brane, nucleus, endolysosomes etc.), organ level (e.g. lung,
spleen, stomach, liver etc.) and/or tissue level (e.g. tumour
cells etc.).
Acknowledgements
The Hassan II Academy of Science and Technology is warmly
acknowledged for the financial support.
Notes and references
1 G. R. Newkome and C. Shreiner, Chem. Rev., 2010, 110, 6338.
2 D. Astruc, E. Boisselier and C. Ornelas, Chem. Rev., 2010,
110, 1857.
3 A. M. Caminade and J. P. Majoral, Chem. Soc. Rev., 2010,
39, 2034.
4 T. D. Patel, B. N. Parikh, G. D. Gothi, J. B. Dave and C. N. Patel,
J. Global Pharma Technol., 2010, 2, 5.
5 M. A. Mintzer and M. W. Grinstaff, Chem. Soc. Rev., 2010,
40, 173.
6 M. Calderón, M. A. Quadir, M. Strumia and R. Haag, Biochimie,
2010, 92, 1242.
7 C. M. Paleos, D. Tsiourvas, Z. Sideratou and L.-A. Tziveleka,
Expert Opin. Drug Delivery, 2010, 7, 1387.
8 J. M. Oliveira, A. J. Salgado, N. Sousa, J. F. Mano and R. L. Reis,
Prog. Polym. Sci., 2010, 35, 1163.
9 A.-M. Caminade, C.-O. Turrin and J.-P. Majoral, New J. Chem.,
2010, 34, 1512.
10 M. Raviña, P. Paolicelli, B. Seijo and A. Sanchez, Mini-Rev. Med.
Chem., 2010, 10, 73.
11 S. Zhu, M. Hong, L. Zhang, G. Tang, Y. Jiang and Y. Pei, Pharm.
Res., 2010, 27, 161.
12 M. Zablocka, A. Hameau, A. M. Caminade and J. P. Majoral,
Adv. Synth. Catal., 2010, 352, 2341.
13 A. M. Caminade, A. Hameau and J. P. Majoral, Chem.–Eur. J.,
2009, 15, 9270.
14 S. H. Medina and M. E. H. El-Sayed, Chem. Rev., 2009, 109, 3141.
15 J. R. Baker Jr., Am. Soc. Hematol., 2009, 708.
16 R. K. Tekade, P. V. Kumar and N. K. Jain, Chem. Rev., 2009,
109, 49.
17 O. Rolland, C.-O. Turrin, A.-M. Caminade and J.-P. Majoral,
New J. Chem., 2009, 33, 1809.
18 G. R. Newkome and C. D. Shreiner, Polymer, 2008, 49, 1.
19 P. Niederhafner, M. Reinis, J. Sebestik and J. Jezek, J. Pept. Sci.,
2008, 14, 556.
20 A. M. Caminade, C. O. Turrin and J. P. Majoral, Chem.–Eur. J.,
2008, 14, 7422.
21 A. M. Caminade, P. Servin, R. Laurent and J. P. Majoral, Chem.
Soc. Rev., 2008, 37, 56.
22 D. A. Tomalia, L. A. Reyna and S. Svenson, Biochem. Soc. Trans.,
2007, 35, 61.
23 Y. Cheng, Z. Xu, M. Ma and T. Xu, J. Pharm. Sci., 2007, 97, 123.
24 E. R. Gillies and J. M. J. Fréchet, Drug Discovery Today, 2005,
10, 35.
25 A. K. Patri, J. F. Kukowska-Latallo and J. R. Baker Jr., Adv. Drug
Delivery Rev., 2005, 57, 2203.
26 T. D. McCarthy, P. Karellas, S. A. Henderson, M. Giannis,
D. F. O’Keefe, G. Heery, J. R. A. Paull, B. R. Matthews and
G. Holan, Mol. Pharmaceutics, 2005, 2, 312.
27 E. E. Simanek and S. O. Gonzalez, J. Chem. Educ., 2002, 79, 1222.
28 A.-M. Caminade and J.-P. Majoral, Acc. Chem. Res., 2004,
37, 341.
29 K. A. Jacobson, Trends Pharmacol. Sci., 2010, 10, 575.
30 J. Ogier, T. Arnauld and E. Doris, Future Med. Chem., 2009,
4, 693.
31 (a) C. M. Paleos, D. Tsiourvas, Z. Sideratou and L.-A. Tziveleka,
Expert Opin. Drug Delivery, 2010, 7, 1387; (b) M. X. Tang,
C. T. Redemann and F. C. Szoka Jr., Bioconjugate Chem., 1996,
7, 703.
32 M. Hayder, M. Poupot, M. Baron, D. Nigon, C.-O. Turrin,
A.-M. Caminade, J.-P. Majoral, R. A. Eisenberg, J.-J. Fournié,
New J. Chem., 2012, 36, 227–240 239

A. Cantagrel, R. Poupot and J.-L. Davignon, Sci. Transl. Med.,
2011, 3, 81.
33 S. Svenson and D. A. Tomalia, Adv. Drug Delivery Rev., 2005,
57, 2106.
34 A. R. Menjoge, R. M. Kannan and D. A. Tomalia, Drug Discovery
Today, 2010, 15, 171.
35 C. Kojima, Expert Opin. Drug Delivery, 2010, 7, 307.
36 W. Gao, J. M. Chan and O. C. Farokhzard, Mol. Pharmaceutics,
2010, 7, 1913.
37 R. X. Zhuo, B. Du and Z. R. Lu, J. Controlled Release, 1999,
57, 249.
38 D. Wang, P. Kopeckova, T. Minko, V. Nanayakkara and
J. Kopecek, Biomacromolecules, 2000, 1, 313.
Published on 08 September 2011. Downloaded by MIT Library on 2/10/2021 12:35:19 AM.
39 S. Zhu, M. Hong, L. Zhang, G. Tang, Y. Jiang and Y. Pei, Pharm.
Res., 2010, 27, 161.
40 S. Gurdag, J. Khandare, S. Stapels, L. H. Matherly and
R. M. Kannan, Bioconjugate Chem., 2006, 17, 275.
41 M. Najlah, S. Freeman, D. Attwood and A. D’Emanuele, Biocon-
jugate Chem., 2007, 18, 937.
42 T. P. Thomas, I. J. Majoros, A. Kotlyar, J. F. Kukowska-Latallo,
A. Bielinska, A. Myc and J. R. Baker Jr., J. Med. Chem., 2005,
48, 3729.
43 I. J. Majoros, T. P. Thomas, C. B. Mehta and J. R. Baker Jr.,
J. Med. Chem., 2005, 48, 5892.
44 J. F. Kukowska-Latallo, K. A. Candido, Z. Cao, S. S. Nigavekar,
I. J. Majoros, T. P. Thomas, T. P. Balogh, M. K. Khan and
J. R. Baker Jr., Cancer Res., 2005, 65, 5317.
45 Y. Zhang, T. P. Thomas, K.-H. Lee, M. Li, H. Zong, A. M. Desai,
A. Kotlyar, B. Huang, M. M. Banaszak Holl and J. R. Baker Jr.,
Bioorg. Med. Chem., 2011, 19, 2557.
46 L. V. Haandel and J. F. Stobaugh, Anal. Bioanal. Chem., 2010,
397, 1841.
47 T. P. Thomas, S. K. Choi, M.-H. Li, A. Kotlyar and J. R. Baker
Jr., Bioorg. Med. Chem. Lett., 2010, 20, 5191.
48 I. J. Majoros, A. Myc, T. Thomas, C. B. Mehta and J. R. Baker Jr.,
Biomacromolecules, 2006, 7, 572.
49 X. Bi, X. Shi, I. J. Majoros, R. Shukla and J. R. Baker Jr.,
J. Comput. Theor. Nanosci., 2007, 4, 1187.
50 S. Tang, L. J. Martinez, A. Sharma and M. Chai, Org. Lett., 2006,
8, 4421.
51 N. Malik, R. Wiwattanapatapee, R. Klopsch, K. Lorenz, H. Frey,
J. W. Weener, E. W. Meijer, W. Paulus and R. Duncan,
J. Controlled Release, 2000, 65, 133.
52 H. R. Ihre, O. L. De Jesus, F. C. Szoka and J. M. J. Fréchet,
Bioconjugate Chem., 2002, 13, 443.
53 S. Jain, A. Kaur, R. Puri, P. Utreja, A. Jain, M. Bhide, R. Ratnam,
V. Singh, A. S. Patil, N. Jayaraman, G. Kaushik, S. Yadav and
K. L. Khanduja, Eur. J. Med. Chem., 2010, 45, 4997.
54 J. Lim and E. E. Simanek, Org. Lett., 2008, 10, 201.
55 O. Perumal, J. Khandare, P. Kolhe, S. Kannan, M. Lieh-Lai and
R. M. Kannan, Bioconjugate Chem., 2009, 20, 842.
56 R. Inapagollaa, B. Raja Gurua, Y. E. Kurtoglua, X. Gaob,
M. Lieh-Laic, D. J. P. Bassett and R. M. Kannan, Int. J. Pharm.,
2010, 399, 140.
57 Y. Zhang, T. P. Thomas, A. Desai, H. Zong, P. R. Leroueil,
I. J. Majoros and J. R. Baker Jr., Bioconjugate Chem., 2010,
21, 489.
58 B. Wang, R. S. Navath, R. Romero, S. Kannan and R. Kannan,
Int. J. Pharm., 2009, 377, 159.
59 R. S. Navath, Y. E. Kurtoglu, B. Wang, S. Kannan, R. Romero
and R. M. Kannan, Bioconjugate Chem., 2008, 19, 2446.
60 H. R. Ihre, O. L. Padilla De Jesus, F. C. Szoka Jr. and J. M. J.
Fréchet, Bioconjugate Chem., 2002, 13, 443.
61 W. C. Floyd, G. K. Datta, S. Imamura, H. M. Kieler-Ferguson,
K. Jerger, A. W. Patterson, M. E. Fox, F. C. Szoka, J. M. J.
Fréchet and J. A. Ellman, ChemMedChem, 2011, 6, 49.
62 R. Wiwattanapatapee, L. Lomlim and K. Saramunee, J. Controlled
Release, 2003, 88, 1.
240 New J. Chem., 2012, 36, 227–240 This journal is
View Article Online
63 F. Alam, A. H. Soloway, R. F. Barth, N. Mafune, D. M. Adams
and W. H. Knoth, J. Med. Chem., 1989, 32, 2326.
64 R. F. Barth, D. M. Adams, A. H. Soloway, F. Alam and
M. V. Darby, Bioconjugate Chem., 1994, 5, 58.
65 C. Wängler, S. Maschauer, O. Prante, M. Schäfer, R. Schirrmacher,
P. Bartenstein, M. Eisenhut and B. Wängler, ChemBioChem, 2010,
11, 2168.
66 T. Darbre and J.-L. Reymond, Acc. Chem. Res., 2006, 39, 925.
67 A. V. Klein and T. W. Hambley, Chem. Rev., 2009, 109, 4911.
68 E. Gabano, M. Ravera and D. Osella, Curr. Med. Chem., 2009,
16, 4544.
69 P. J. Pellechia, J. Gao, Y. Gu, H. J. Ploehn and C. J. Murphy,
Inorg. Chem., 2004, 43, 1421.
70 N. Malik, E. G. Evavgoraou and R. Duncan, Anti-Cancer Drugs,
1999, 10, 767.
71 I. Haririan, M. S. Alavidjeh, M. R. Khorramizadeh, M. S. Ardestani,
Z. Z. Ghane and H. Namazi, Int. J. Nanomed., 2010, 20, 63.
72 E. Bellis, L. Hajba, B. Kovacs, K. Sandor, L. Kollar and
G. Kokotos, J. Biochem. Biophys. Methods, 2006, 69, 151.
73 X. Zhao, S. C. J. Loo, P. P.-F. Lee, T. T. Y. Tan and C. K. Chu,
J. Inorg. Biochem., 2010, 104, 105.
74 W.-D. Jang, N. Nishiyama, G.-D. Zhang, A. Harada, D.-L. Jiang,
S. Kawauchi, Y. Morimoto, M. Kikuchi, H. Koyama, T. Aida and
K. Kataoka, Angew. Chem., Int. Ed., 2005, 44, 419.
75 J. Kim, H.-J. Yoon, S. Kim, K. Wang, T. Ishii, Y.-R. Kim and
W.-D. Jang, J. Mater. Chem., 2009, 19, 4627.
76 A. Quintana, E. Raczka, L. Piehler, I. Lee, A. Myc, I. Majoros,
A. K. Patri, T. Thomas, J. Mulé and J. R. Baker Jr., Pharm. Res.,
2002, 19, 1310.
77 C. C. Lee, E. R. Gillies, M. E. Fox, S. J. Guillaudeu, J. M. J.
Fréchet, E. E. Dy and F. C. Szoka, Proc. Natl. Acad. Sci. U. S. A.,
2006, 103, 16649.
78 M. Najlah, S. Freeman, D. Attwood and A. D’Emanuele, Int. J.
Pharm., 2006, 308, 175.
79 K. Kono, C. Kojima, N. Hayashi, E. Nishisaka, K. Kiura,
S. Watarai and A. Harada, Biomaterials, 2008, 29, 1664.
80 J. Lim, A. Chouai, S.-T. Lo, W. Liu, X. Sun and E. E. Simanek,
Bioconjugate Chem., 2009, 20, 2154.
81 Y. E. Kurtoglua, M. K. Mishraa, S. Kannanb and R. M. Kannan,
Int. J. Pharm., 2010, 384, 189.
82 R. S. Navath, A. R. Menjoge, B. Wang, R. Romero, S. Kannan
and R. M. Kannan, Biomacromolecules, 2010, 11, 1544.
83 D. A. Patel, J. E. Henry and T. A. Good, Biochim. Biophys. Acta,
Gen. Subj., 2006, 1760, 1802.
84 D. A. Patel, J. E. Henry and T. A. Good, Brain Res., 2007,
1161, 95.
85 Y. Y. Jiang, G.-T. Tang, L.-H. Zhang, S.-Y. Kong, S.-J. Zhu and
Y.-Y. Pei, J. Drug Targeting, 2010, 18, 389.
86 Y. Cheng and T. Xu, Eur. J. Med. Chem., 2008, 43, 2291.
87 S. N. Patel, P. Sethi, K. Ansari, D. Tiwari, A. Sharma and
A. K. Singhai, Int. J. Pharm. Life Sci., 2010, 1, 382.
88 R. Haag and F. Kratz, Angew. Chem., Int. Ed., 2006, 45, 1198.
89 R. Satchi-Fainaro, R. Duncan and C. M. Barnes, Adv. Polym. Sci.,
2006, 193, 1.
90 Y. Deguchi, A. Kurihara and W. M. Pardridge, Bioconjugate
Chem., 1999, 10, 32.
91 E. V. Batrakova, T. Y. Dorodnych, E. Y. Klinkii,
E. N. Kliushnenkova, O. B. Shemchukova, O. N. Goncharova,
S. A. Arjakov, V. Y. Alakhov and A. V. Kabanov, Br. J. Cancer,
1996, 74, 1545.
92 T. Nakanishi, S. Fukushima, K. Okamoto, M. Suzuki,
Y. Matsumura, M. Yokayama, T. Okano, Y. Sakarai and
K. Kataoka, J. Controlled Release, 2001, 74, 295.
93 S. Hong, P. R. Leroueil, I. J. Majoros, B. G. Orr, J. R. Baker and
M. M. B. Holl, Chem. Biol., 2007, 14, 107.
94 M. Najlah and A. D’Emanuele, Curr. Opin. Drug Discovery Dev.,
2007, 10, 756.
95 S. Svenson, Eur. J. Pharm. Biopharm., 2009, 71, 445.
c The Royal Society of Chemistry and the Centre National de la Recherche Scientifique 2012