s targeted!nanopar · i e s for p, c es d b s peo oh + ch 3 o ε-cl peo noh + n 3 ε-cl 3 synthesis...
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Synthesis of func.onalized amphiphilic copolymers for the realiza.on of mul.-‐
targeted nanopar.cles by
M. Malinconico1, G. Dal Pogge1o1, A. Romanelli2, F. Quaglia2, P. Laurienzo1
1 Ins&tute for Polymers, Composites and Biomaterials, IPCB-‐CNR, Pozzuoli 2 Department of Pharmacy, University of Naples Federico II, Naples, Italy
e-‐mail: mario.malinconico@ipcb.cnr.it
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coordinated by:
Drug Delivery Laboratory, Department of Pharmacy, University of Napoli Federico II, Naples, Italy
in collabora4on with:
Department of Biology, University of Padova, Padua, Italy InsItute of Biostructure and Bioimaging, CNR, Naples, Italy
IG 2014 15764
The activities are performed in the framework of the project “Strategies to drive chemotherapeutics to solid tumors: multifunctional nanoparticles from tailor-made block copolymers” granted by AIRC-2014.
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+PEO OH
ε-CL CH3O
+PEO OH
ε-CL N3 N3
Synthesis of functionalized self-assembling amphiphilic copolymers
“Click” Chemistry
PEO-PCL
NPs decorated with peptides containing ‘’CendR’’ motif are able to accumulate at tumor level as they undergo transport pathway in vascular endothelial cells of tumor vessels
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Active targeting
Vascular targeting
Passive targeting
Our general strategy is to design double-‐targeted NPs exposing on the surface both Tumor Penetra&ng Pep&des and folate groups binding folate receptor, which is over-‐expressed on the vast majority of cancer &ssues while its expression is limited in healthy &ssues. Furthermore, NPs slowly release their drug cargo in the body once homing at tumor level is achieved.
• Biomime&c • Enhanced transport through tumor vasculature and improved distribu&on inside tumor inters&&um through decora&on with RGD and CendR mo&fs
• Enhanced accumula&on in cancer cells through folate decora&on
• Sustained docetaxel release in tumor inters&&um
AIM
Amphiphilic diblock copolymers
Strategy
o PCL-‐b-‐PEG with op&mal block length and hydrophilic/lipophilic balance providing NP stability in the blood and long circula&on;
o PCL-‐b-‐PEG-‐TPP modified at PEG site through covalent linkage of already known or novel pepIde sequences that promote specific transport of NPs through tumor vasculature and accumula&on in the tumor;
o PCL-‐b-‐PEG-‐Fol modified at PEG site with folate that promote NP uptake in cancer cells via receptor-‐mediated endocytosis.
PCL
PEO
PepIde (NGR, CendR)
Folate
PEG2000-‐b-‐PCL4000
Fol –PEG2000-‐b-‐PCL4000
TPP–PEG2000-‐b-‐PCL4000 Docetaxel
DBLS/DBL-Fol
DBLL/DBL-Fol PCL-PEGL PCL-PEGS
DBLS DBLL
PCL-PEGS +PCL-PEG-Fol*
PCL-PEGS +PCL-PEG-Fol*
* 4:1 wt ratio
PCL4.7-PEG1.5-Fol
PCL4.7-PEG1.0
PCL4.7-PEG2.0
PCL-PEGS
PCL-PEGL
PCL-PEG-Fol
Om
OO
O
OHn
CH3
OHO
NH
O
NH
N
ONH
NH2N
NO
HNN
NN
m
OO
O
OHn
PCL-PEG1000
PCL-PEG1500-Fol
PCL-PEG2000
PCL-PEG1500-Fol
DBL1000 DBL2000
Folate exposi&on
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mPEG1000-OHO
O
mPEG1000-PCL4000
N3-PEG1500-OHO
O
N3-PEG1500-PCL4000
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Step ISO
OCl S
O
O
NaN3
Step II
O
O
Step III
OH-PEG1,5k-OH O-PEG1,5k-OH
SO
OO-PEG1,5k-OH N3-PEG1,5k-OH
N3-PEG1,5k-OH N3-PEG1,5k-PCL4k-OH
HETERO BUILDING BLOCK
Ag2O, KI
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TAGS
FOLIC ACID
PEPTIDE
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FOLATE HPLC MASS
O OH
NH
O
NH
NO
HN
H2N N N
O
OH
NH2
O OH
NH
O
NH
NO
HN
H2N N N
O
HN
O
NH
O
NH
NO
HN
H2N N N
O
OH
NH
γ
α
propargylamine
X
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PCL-PEG-FOL SYNTHESIS O OH
NH
O
NH
NO
HN
H2N N N
O
NH
O OH
NH
O
NH
NO
HN
H2N N N
O
HN
N NN PEG1,5k-PCL4k-OH
α
γ
N3-PEG1,5k-PCL4k-OHγ
α
N3-‐PEG1.5k-‐PCL4k FOL-‐PEG1.5k-‐PCL4k
_
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TEM image of a single DBLS/DBL-Fol SEM images of DBLS/DBL-
Fol freeze-dried
NPs were prepared by solvent diffusion of an organic phase added dropwise in water under magnetic stirring. After solvent evaporation, NPs were filtered, freeze-dried and stored at 4 °C.
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CONFOCAL MICROSCOPY KB cells, 1 h pre-incubation 1 mM free fol 1 h NP incubation (20 µg/ml NPs) in medium + 10% serum
PEG1000 PCL4000
Free fol + PEG1000 PCL4000 Free fol + 20% fol PEG1000 PCL4000
20% fol PEG1000 PCL4000
Effective signal reduction
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Future developments:
POLYROTAXANE
polymer+cd+endcapping
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Synthesis of funcIonalized amphiphilic copolymers for the realizaIon of mulI-‐targeted nanoparIcles
thanks for the a:en.on!
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