TY - JOUR
T1 - Novel Oligo-Guanidyl-PEG Carrier Forming Rod-Shaped Polyplexes
AU - Malfanti, Alessio
AU - Mastrotto, Francesca
AU - Han, Yanxiao
AU - Král, Petr
AU - Balasso, Anna
AU - Scomparin, Anna
AU - Pozzi, Sabina
AU - Satchi-Fainaro, Ronit
AU - Salmaso, Stefano
AU - Caliceti, Paolo
N1 - Publisher Copyright:
© 2019 American Chemical Society.
PY - 2019/4/1
Y1 - 2019/4/1
N2 - A novel unconventional supramolecular oligo-cationic structure (Agm 6 -M-PEG-OCH 3 ) has been synthesized to yield high efficiency therapeutic oligonucleotide (ON) delivery. Agm 6 -M-PEG-OCH 3 was obtained by a multistep protocol that included the conjugation of agmatine (Agm) moieties to maltotriose (M), which was further derivatized with one poly(ethylene glycol) (PEG) chain. Gel electrophoresis analysis showed that the 19 base pairs dsDNA model ON completely associates with Agm 6 -M-PEG-OCH 3 at 3 N/P molar ratio, which is in agreement with the in silico molecular predictions. Isothermal titration calorimetry (ITC) analyses showed that the Agm 6 -M-PEG-OCH 3 /ON association occurs through a combination of mechanisms depending on the N/P ratios resulting in different nanostructures. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) revealed that the Agm 6 -M-PEG-OCH 3 /ON polyplexes have rod-shape structure with a mean diameter of 50-75 nm and aspect ratio depending on the N/P ratio. The polyplexes were stable over time in buffer, while a slight size increase was observed in the presence of serum proteins. Cell culture studies showed that neither Agm 6 -M-PEG-OCH 3 nor polyplexes displayed cytotoxic effects. Cellular uptake depended on the cell line and polyplex composition: cellular internalization was higher in the case of MCF-7 and KB cells compared to MC3T3-E1 cells and polyplexes with smaller aspect ratio were taken-up by cells more efficiently than polyplexes with higher aspect ratio. Finally, preliminary studies showed that our novel carrier efficiently delivered ONs into cells providing gene silencing.
AB - A novel unconventional supramolecular oligo-cationic structure (Agm 6 -M-PEG-OCH 3 ) has been synthesized to yield high efficiency therapeutic oligonucleotide (ON) delivery. Agm 6 -M-PEG-OCH 3 was obtained by a multistep protocol that included the conjugation of agmatine (Agm) moieties to maltotriose (M), which was further derivatized with one poly(ethylene glycol) (PEG) chain. Gel electrophoresis analysis showed that the 19 base pairs dsDNA model ON completely associates with Agm 6 -M-PEG-OCH 3 at 3 N/P molar ratio, which is in agreement with the in silico molecular predictions. Isothermal titration calorimetry (ITC) analyses showed that the Agm 6 -M-PEG-OCH 3 /ON association occurs through a combination of mechanisms depending on the N/P ratios resulting in different nanostructures. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) revealed that the Agm 6 -M-PEG-OCH 3 /ON polyplexes have rod-shape structure with a mean diameter of 50-75 nm and aspect ratio depending on the N/P ratio. The polyplexes were stable over time in buffer, while a slight size increase was observed in the presence of serum proteins. Cell culture studies showed that neither Agm 6 -M-PEG-OCH 3 nor polyplexes displayed cytotoxic effects. Cellular uptake depended on the cell line and polyplex composition: cellular internalization was higher in the case of MCF-7 and KB cells compared to MC3T3-E1 cells and polyplexes with smaller aspect ratio were taken-up by cells more efficiently than polyplexes with higher aspect ratio. Finally, preliminary studies showed that our novel carrier efficiently delivered ONs into cells providing gene silencing.
KW - cationic carriers
KW - nanoparticles
KW - nonviral carriers
KW - oligonucleotides delivery
KW - polyplexes
UR - http://www.scopus.com/inward/record.url?scp=85063535285&partnerID=8YFLogxK
U2 - 10.1021/acs.molpharmaceut.9b00014
DO - 10.1021/acs.molpharmaceut.9b00014
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AN - SCOPUS:85063535285
VL - 16
SP - 1678
EP - 1693
JO - Molecular Pharmaceutics
JF - Molecular Pharmaceutics
SN - 1543-8384
IS - 4
ER -