Supramolecular Lipid Nano Assemblies of PEGMA-co-Stearic Acid Block

Copolymer via RAFT Polymerization for Enhanced Anti-Cancer Drug Delivery

Nayan Chakraborty,Priyatosh Sarkar,Rima Saha,Kishor Sarkar*
Gene Therapy and Tissue Engineering Lab (GTTL), Department of Polymer Science and Technology,
University of Calcutta, 92, A.P.C. Road, Kolkata-700009, India
*Email- kishorpst@gmail.com/ kspoly@caluniv.ac.in

Abstract:In pursuit of innovative solutions for cancer therapy,we present a novel approach to drug delivery through the creation of supramolecular lipid nano-assemblies.This strategy leverages reversible addition-
fragmentation chain transfer (RAFT) Polymerization to engineer core-shell polymer structures with distinct hydrophilic and hydrophobic regions.The hydrophobic segment is crafted from stearic acid,while the
hydrophilic counterpart is derived from polyethylene glycol methyl ether methacrylate (PEGMA).Through controlled RAFT polymerization,these components are skillfully combined to yield the final block
copolymer,poly (PEGMA-co-Stearic Acid) BCP.These versatile BCPs serve as the foundation for our advanced drug delivery platform,which we have designed to enhance the efficacy of anti-cancer medications.To
demonstrate the potential of this delivery system,we loaded these BCPs with doxorubicin (DOX),a well-established anti-cancer drug.The results were remarkable,showcasing the controlled release of DOX in an acidic
environment,mirroring the Ph conditions prevalent in cancerous tissues.Furthermore,our investigations revealed significant cellular uptake of DOX-loaded BCPs by MDA-MB-231 triple-negative breast cancer
cells.This uptake led to a remarkable 35-fold increase in anti-cancer activity compared to free DOX.The efficacy of this approach was further validated through cellular apoptosis,as observed through scanning electron
microscopy (SEM) imaging.Our findings suggest that these supramolecular lipid nano-assemblies represent a highly promising avenue for the enhanced and targeted delivery of anti-cancer drugs.By harnessing the
power of RAFT polymerization and utilizing PEGMA-co-Stearic Acid BCPs,we offer a potential breakthrough in cancer therapy,with the potential to significantly improve treatment outcomes.

Recent Cancer statistics Schematic representation of Characterization of polymers

synthesis of PEGMA-co-SEMA
block co-polymer

1H NMR spectra of synthesized-

(a) poly(PEGMA)
(b) poly(PEGMA-co-SEMA)(BCP50) polymer and
(c) FTIR spectra of poly(PEGMA) and poly(PEGMA-
Graphical representation of the synthesis co-SEMA) (BCP50) polymer.

TEM & size distribution of BCP

Fig.(a) shows the regular shape nanoparticle

morphology of BCP50 and
(b) shows the size distri-bution curve where
maximum nanoparticles are within size range from
90–100 nm

Drug release and Cellular uptake Confocal microscopy images

(b)

Mass ratio (polymer : DOX) DOX feed (mg) DL% EE%

BCP10 1 : 0.1 10 2.53

BCP25 1 : 0.5 10 5.1

BCP50 1:1 10 5.98

Physical entrapment of hydrophobic DOX to a hydrophobic core is dependent
upon the hydrophobe chain length & the quantity
It is found that DOX entrapment is proportional with the increase of stearic acid
content i.e.,BCP10 shows only 2.53% drug loading & BCP50 shows higher drug
loading capability
Confocal microscopy images of fixed MDA MB-231 cells incubated with BCP10-DOX, BCP25-DOX
and BCP50-DOX for 4 hours
(a)–(c) corresponds to the stained nucleus by DAPI
Similarly, (a1)–(c1) are the corresponding confocal images of cells which shows successful uptake
of DOX loaded nanoparticle
(a2)–(c2) are the bright field images of the corresponding cells
(a3)–(c3) are the merged images where purple colour shows co-localization of the blue and red
colour due to DAPI and DOX fluorescence which suggests successful uptake of the DOX loaded
nanoparticle
(a4)–(c4) are the 3D interactive plots of intracellular DOX and DAPI fluorescence and (a5)–(c5) are
the measured intra- cellular DOX intensity

Conclusion: A core-shell nano template have been designed by

Cell viability assay & SEM study using PEGMA and SEMA via RAFT polymerization for the
delivery of DOX to MDA MB-231 cancer cell. Thus, wrapping them
in an amphiphilic polymer with a lipid segment may thereby
increase their biodistribution and access to the cancer environment.
These results suggested that in near future, BCP50 could be a
suitable nanomaterial for hydrophobic anticancer drug delivery
systems.

References:
•G. Kwon, S. Suwa, M. Yokoyama, T. Okano, Y. Sakurai and K. Kataoka, J. Controlled Release,
1994
(a) Cell viability assay of BCP10, BCP25 and BCP50 using MDA-MB-231 •M. J. Joralemon, S. McRae and T. Emrick, Chem. Commun., 2010, 46, 1377–1393
Cellular morphology under SEM where,
cell line (a) control (without treatment)
•R. Saha, S. Bhayye, S. Ghosh, A. Saha and K. Sarkar, ACS Appl. Bio Mater., 2019, 2, 5349–5365
(b) MTT assay of free DOX and DOX loaded P(PEGMA-co- SEMA) block
(b) DOX treated and
co-polymer (BCP10-DOX, BCP25-DOX, BCP50-DOX) (c) BCP50-DOX treated cells
(c) determined 50% inhibitory concentration (IC50) value of DOX, BCP10-
DOX, BCP25-DOX, BCP50-DOX
Acknowledgement:
The work was financially supported by DST-SERB. Thanks