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Materials Chemistry B
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FEATURE ARTICLE
sensitive drug release, incorporation of agents for imaging and the attachment of ligands for active
targeting. By putting it all together, it may be possible to obtain an ideal multifunctional nanocarrier for
cancer therapy. Among the many eorts made so far to obtain one, SLN/NLC should have a place in this
DOI: 10.1039/c3tb20990c
search for a combined therapeutic and diagnostic system with dramatically enhanced ecacy in cancer
www.rsc.org/MaterialsB
therapy.
Introduction
Table 1
Feature Article
New trends with lipid nanocarriers for enhanced tumor activity and diagnosis
Formulation
Drug/agent
Added function
Reference(s)
SLN
SLN
SLN
SLN
SLN
NLC
SLN
SLN
PEGylation
PEGylation
PEGylation
PEGylation
Co-loading
Co-loading
Co-loading
Co-loading
15 and 19
21 and 22
23
24
29
30
31 and 32
33
Co-loading/pegylation/gene delivery
Co-loading/pH1 stimulus-sensitive
pH stimulus-sensitive
pH stimulus-sensitive
34
35
7
38
Imaging diagnostic
Imaging diagnostic
Imaging diagnostic
PEGylation/co-loading/imaging
diagnostic/gene delivery
Active targeting/imaging diagnostic
39
40
42
41
Active targeting
Active targeting
Active targeting
Active targeting/PEGylation/gene delivery
Active targeting/PEGylation/gene delivery
Active targeting/PEGylation
45
46
47
48
49
50
Active targeting/PEGylation
51
Active targeting/PEGylation/co-loading
Active targeting/PEGylation/ph stimulus-sensitive
Active targeting
52
53
59
SLN
SLN
SLN
SLN
SLN
SLN
SLN
SLN
SLN
SLN
SLN
SLN
SLN
SLN
NLC
SLN
SLN
NLC
NLC
and NLC loaded with doxorubicin were 2.5 more eective than
the respective free drugs.9,10 SLN loaded with docetaxel were
shown to be more eective than the commercial docetaxel,
Taxotere, in vitro against colorectal (C-26) and malignant
melanoma (A-375) cell lines. In in vivo tests with C-26-implanted
BALB/c mice, SLN loaded with docetaxel showed better tumor
inhibitory ecacy and survival than Taxotere.11
SLN carrying dierent antineoplastic agents (doxorubicin,
paclitaxel, cholesteryl butyrate, and anti-VEGF antisense oligonucleotides) were tested in experimental animal models of
cerebral gliomas and induced an eective anti-tumoral therapeutic response.12 An explanation involves the accumulation of
drugs encapsulated in SLN in brain aer intravenous administration. This was attributed to the transport of intact SLN
through the bloodbrain barrier and also to the sustained
release from the lipid matrix.12,13,14 Pharmacokinetics studies
showed a prolonged circulation up to 24 hours, a signicant
increase in t1/2, Cmax and low uptake of SLN by the reticuloendothelial system (RES). A lower concentration of doxorubicin
was found in heart when administered as SLN, indicating the
possibility of lower cardiotoxicity compared to the commercial
doxorubicin solution, which is one of the eects limiting the
usefulness of doxorubicin.15
31
Thus, SLN and NLC can oer many benets for cancer
therapy. Recent works have described new proposals to further
optimize chemotherapy using these drug delivery systems. They
have been successfully multifunctionalized to capture a payload
of drugs, to target specic cells and to release entrapped drugs
in a controlled manner. The following topics of this review focus
on these new trends, related to the addition of more functions
to these nanoparticles, such as PEGylation, co-encapsulation of
anticancer agents, pH-sensitivity, magnetic nanoparticle
loading and other agents for imaging and for attachment of
ligands on the surface (see Table 1).
Feature Article
tumors due to the defective vasculature, resulting in a passive
targeting. This is a well-established phenomenon, termed the
enhanced permeability and retention (EPR) eect.16,17 Poly(ethylene)glycol (PEG) derivatives and others hydrophilic polymers have been widely employed to modify nanoparticles
surface. This modication results in a steric stabilization of the
carriers and reduction of their uptake by the RES cells. The
steric stabilization is a phenomenon in which the solid
particulates are protected from interactions with dierent
solutes and this term has been used to describe the polymermediated protection.18
Drug-free PEGylated SLN and non-PEGylated SLN were
prepared and injected intravenously in rats to study their tissue
distribution and transport across the bloodbrain barrier.19 The
average size of these lipid nanoparticles was below 100 nm. To
PEGylate the SLN, stearic acid-PEG 2000 at 0.15% w/v was
added. Localization of SLN toward in the brain and in the
cerebrospinal uid (CSF) was higher for both formulations
comparing to the free drug. However, there was a lower uptake
in the liver and lung for PEGylated-SLN than for the nonPEGylated-SLN, conrming the inuence of PEGylation.19
Another study investigated the potential of the same PEGylated
and non-PEGylated SLN for improved parenteral delivery of
doxorubicin.20 The pharmacokinetics and tissue distribution of
doxorubicin in these SLN were compared with a commercial
solution of doxorubicin. There was a signicantly prolonged
circulation, an increase of t1/2 and of Cmax with PEGylated SLN.20
The inuence of stearic acid-PEG 2000 concentration for these
SLN was also evaluated. The AUC of doxorubicin increased as a
function of the amount of PEGylating agent present in the SLN.
Doxorubicin was detected in blood 6 hours post-injection from
both non-PEGylated and PEGylated SLN, while no doxorubicin
was detectable aer intravenous injection of doxorubicin solution. The highest concentration of doxorubicin in the blood was
detected in PEGylated SLN at the highest concentration of
stearic acid-PEG 2000.15
The PEGylation of SLN also increased their cytotoxicity and
uptake for some cancer cell lines. PK-L4 was synthesized as a
potential anticancer compound. IC50 values were signicantly
lower for PK-L4-loaded SLNs with stearic acid-PEG 2000 in the
human lung carcinoma cell line (A549) and a human hepatocellular liver carcinoma cell line (HepG2). This eect was
strongly connected to the SLN incorporation of PEG and was
attributed to the degradation delay using stabilizing surfactants
such as stearic acid-PEG 2000. However, further investigations
about the mechanism need to be done. There was a signicantly
higher uptake of the PK-L4-loaded SLN PEGylated with stearic
acid-PEG 2000 at 10 wt% than with the plain SLN, but the
cytotoxicity was lower and the IC50 of the PEGylated SLN was
higher than with plain SLN.21,22
Long-circulating SLN made with two dierent surfactants,
Pluronic F68 (also known as poloxamer) and Brij78, revealed
marked dierences in pharmacokinetic parameters, especially
in the t1/2 and AUC, in comparison with Cremophor EL
containing a solution of paclitaxel.23 F68 SLN and Brij 78-SLN
were long-circulating with a 7.4-fold and 3.6-fold higher t1/2,
respectively. This was attributed to the reduced uptake of SLN
Feature Article
with paclitaxel. In KB cell-xenograed mice, this SLN loaded
with paclitaxel and complexed with siMCL1 signicantly
reduced the growth of tumors. Thus, it veried the potential of
SLN for the co-delivery of lipophilic anticancer drug and therapeutic siRNA.
Our previous work describes a new SLN co-loaded with
doxorubicin and docosahexaenoic acid (DHA), an omega 3
polyunsaturated fatty acid that shows synergic eect with
anthracyclines.35 Since doxorubicin is a water soluble drug, its
encapsulation in a matrix lipid is challenging. We proposed
the formation of an ionic pair between the cationic doxorubicin molecule and the anionic DHA in situ, during the SLN
preparation, to improve drug encapsulation and anticancer
activity. The formulation had a size of about 90 nm, entrapment eciency 100% and drug loading of 31 mg g1 for
doxorubicin. In vitro evaluation showed higher uptake for
drugs encapsulated in SLN compared to the free drugs as well
as higher cytotoxicity against the human lung cancer cell line,
A549.35
Feature Article
Fig. 1 Schematic of doxorubicin release from SLN at lowered pH. The high encapsulation of doxorubicin into the lipid matrix was achieved by an in situ ionic pair
formation that resulted in a lipophilic salt [(1) DOX-DHA Ionic pair], as previously described in ref. 35. At low pH of 5, the DHA is protonated (2) and will undo the ionic
interaction with doxorubicin (3). Due to the hydrophilic character of the cationic doxorubicin, its diusion from the lipid matrix would be faster and result in enhanced
drug release (4).
Feature Article
SLN were more cytotoxic and were preferably taken up by A549
tumor cells. Pharmacokinetic studies revealed improved
bioavailability, longer half-life and increased mean residence
time of doxorubicin upon mannose conjugation. They also
veried a higher concentration of doxorubicin in tumors for the
mannosylated-SLN as well as lower damage to renal and hepatic
tissues. Thus, the attachment of the small molecule mannose to
the SLN surface seems to be promising to target cancer cells.
However, studies concerning the higher ecacy in vivo of this
formulation have to be investigated.
SLN loaded with paclitaxel and modied with octaarginine
(R8-PTX-SLN) were proposed by ref. 46. The octaarginine was
attached to the SLN through a linkage with stearic acid. The
results showed a higher cytotoxicity against human lung cancer
cells (A549) and an increase of the cell uptake. The authors of
ref. 47 developed SLN loaded with doxorubicin and the antiepithelial growth factor receptor, anti-EGFR, which was electrostatically attached to the catanionic (cationic and anionic)
surface of the particle. Immunochemical staining revealed that
the crosslinked anti-EGFR on the surface of SLN preserved a
high specicity for EGFR on U87MG cells, and it induced a
growth-inhibition eect. Another study proposed a linkage of
transferrin (Tf) on polyethylene glycol-phosphatidylethanolamine (PEG-PE) to obtain Tf-PEG-PE ligands. This polymer was
used for the surface modication of a cationic SLN for dual gene
(plasmid pEGFP) and dexamethasone delivery. In vitro and in
vivo studies showed increase in transfection due to the attachment of Tf.48
To target cancer cells and macrophages in the liver, SLN were
modied with dual ligands Tf and mannan (M). Tf and M were
linked to polyethylene glycol-phosphatidylethanolamine (PEGPE) and PE separately to get transferrin-PEG-PE (T-PEG-PE) and
mannan-PE (M-PE) ligands for the surface modication of SLN.
The in vivo transfection eciency was veried using uorescence protein plasmid (GFP) that was promoted in the presence
of the dual ligands. The evaluation in tumor-bearing animal
models showed that Tf-PEG-PE and M-PE functioned as excellent ligands to improve the cell targeting ability of the carriers.49
A folate-poly(PEG-cyanoacrylate-co-cholesteryl cyanoacrylate)
(FA-PEG-PCHL) was synthesized to modify docetaxel-loaded
NLC and to obtain a long blood circulating eect and improved
targeting.50 This is an amphiphilic copolymer able to locate at
the nanoparticle interface, exposing the PEG chain to the
aqueous phase with the folate moiety at the end. In vivo studies
indicated signicant dierences in the pharmacokinetic prole
and tissue distribution for FA-PEG-PCHL-modied docetaxelloaded NLC compared to the plain docetaxel-loaded NLC. The
PEGylation contributed to an increase of the AUC, t1/2 and MRT,
and there was a signicant accumulation of docetaxel in tumors
due to the folic acid attachment. The authors of ref. 51 veried
that the modication of paclitaxel-loaded SLN with folatepoly(ethylene glycol) (PEG)-phosphatidylethanolamine exhibited a higher tumor inhibition rate in mice with sarcoma180
ascites tumors. It was proposed the co-loading of docetaxel and
ketoconazole in SLN graed with folic acid, which was incorporated by a stearic acid and folic acid conjugate (SA-FA).52 Both
drugs are metabolized hepatically by the cytochrome P-450
Feature Article
system, and ketoconazole can inhibit p-gp eux of docetaxel at
the blood brain barrier. Thus, a potential drugdrug interaction
for these agents can be useful. The authors veried that the
combination of drugs in SLN plus attached folic resulted in
increased brain uptake of docetaxel and it was 44 times higher
than the commercial Taxotere, which suggests promise for
brain tumor treatment.
NLC loaded with paclitaxel were modied with tumor
microenvironment-sensitive polypeptides (TMSP) and folate.53
The aim of the ligands combination was to create a more
selective and ecient system to deliver the drug in cancer cells.
The TMSP and folate were conjugated to the distal ends of
DSPE-PEG2000-maleimide and DSPE-PEG5000-amine, respectively (see Fig. 2). The folate was attached to increase the
endocytosis by the higher interaction with folate-receptors,
which are overexpressed in some tumors. TMSP are shielded
in the blood circulation but are activated by matrix
Fig. 2 (A) A schematic of the dual-ligand DTX-NLC simultaneously modied with folate and TMSP (F/TMSP-DTX-NLC). (B) The schematic of the multivalent interactions
of F/TMSP-DTX-NLC with cancer cells. Reprinted from ref. 53, Copyright (2013), with permission from Elsevier (License number 3133680405511).
Feature Article
functions mentioned. However, the combination of all functions
would be very challenging. Much eort has been done with
liposomes and polymeric nanoparticles to obtain multifunctional nanocarriers. However, the advantages of SLN and NLC
over the others nanosystems qualify them as promising candidates in the development of combined therapeutic and diagnostic multifunctional pharmaceutical nanocarrier systems.
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