RATIONAL CHEMICAL MULTIFUNCTIONALIZATION OF
GRAPHENE INTERFACE ENHANCES TARGETED
CANCER THERAPY
M.A LUCHERELLI, G.REINA, A.BIANCO,
Y.YU, E.MIYAKO, G.ABELLÀN
PRESENTED BY–
SHIVIKA
MSC (USHS) CHEMISTRY SEM 4
27802109319
ANGEW.CHEM. PUBLISHED IN 2020

CANCER :
 Cancer specifically refers to new growth which has
ability to spread to sorrounding tissues which may
eventually lead to death.
 Cancer is the leading cause of death worldwide,
accounting for nearly 10 millions of death in 2020.
 The most common cancers are breast, lung, colon
and rectum or prostate cancer.
 It can be cured if detected early and treated
effectively.

TRADITIONAL WAYS TO TREATING CANCER :
CHEMOTHERAPY:
 Use of anti-cancer drugs to destroy cancer
cells.
 Works by disrupting the growth of cancer
cells.
RADIATION THERAPY:
Use in 2 different ways:
 To cure cancer by destroying tumors.
 To reduce symptoms by reducing size of a
tumor.

NANOTECHNOLOGY:
 Nanotechnology is rapidly evolving domain.
 In the last decades, The interest on the applications of nanomaterials in
Nano medicine has increased Rapidly.
 Large varieties of reactions and their properties like specific cell
targeting , controlled drug release, photothermal and photodynamic
therapies and imaging. Targeted drug delivery approach minimize drug-
originated systemic toxic effects.
 A great advantage is their ability to complex a high quantity of
hydrophobic drugs and improving the uptake and release of drug and
improving the selectivity of free drug.
 Properties like biocompatibility, amount of drug loading, interaction
with biological environment and the biodistribution into the body after
administration.

GRAPHENE :
 Graphene is an allotrope of carbon consisting of a sheet of 2D single
layer sp2 hybridized C atoms.
 Each C atom is attached to it’s 3 nearest neighbors C atoms in same
plane. Thinnest known material.
 Graphene has been found to have good biocompatibility and
biodegradability.
 Moreover, it’s high surface/mass ratio allows a high drug loading , by
absorbing organic molecule through π-π stacking.
 Due to easy functionalization and good water dispersibility , GO has
been widely studied as drug carrier .
 But graphene presents a higher chemical stability wrt GO to reductive
conditions and fewer interaction with cell membranes.

MULTIFUNCTIONAL GRAPHENE PLATFORM
(MFG):
 Covalent multifunctional Graphene platform (mfG).
 Focus on targeting , tracking , delivery and photothermal activity of mfg on cancer cells .
 We selected ICG as fluorophore , FA as targeting agent , DOX as therapeutic molecule.
 The starting material to prepare this Platform corresponds to a multifunctional Graphene which is obtained by
Covalent multifunctionalization of KC8 with diazonium salts

BOC : 3° butyloxycarbonyl
CBZ : benzyloxycarbonyl
Indocyanine green : ICG
PHT : Pthalamide
MOLECULAR STRUCTURES :
Doxorubicin : DOX

SYNTHESIS OF NEW TARGETED DRUG
DELIVERY CONJUGATE :
 The order of introduction of new functionalities is crucial.
 The chemical stability of each functional group under the
deprotection conditions of each amine are initially
investigated.
 The PEG(Polyethylene glycol) Chain plays 2 important roles :
1) To Increase water dispersibility and biocompatibility and 2)
To act as a spacer for linking FA to conjugate.
 The characteristics of imine bond is its reversibility in acidic
medium , at PH<5.5.
 The low rxn yield due to steric hindrance caused by PEG
chain and the other functionalities on the Surface of
graphene.

THERAPEUTIC POTENTIAL :
 Water – soluble tetrazolium (WST) – based assay.
 48 hr of incubation at increasing doses – a slight cell
mortality was observed for Hela cells.
 2 factors : low water dispersibility and low amount of
DOX .
 SOLUTION : we sonicated the prepared mfG/PEG-
FA/ICG/DOX in a BSA and Dox Solution.
 Drug release efficiency in neutral and acidic conditions.
 Evaluated again the in vitro performance.
 Selective anti-cancer activity mediated by folate receptor
– directed targeting effect.

DOX- COMPLEXED AND FOLIC ACID –FUNCTIONALIZED-MFG
GRAPHENE NANOSHEETS COULD BE USED AS AN EFFECTIVE
NANOCARRIER FOR SELECTIVE CANCER KILLING.

CELLULAR UPTAKE BEHAVIOUR OF mfg
NANO CONJUGATES :
 Hela cells were incubated with ICG labelled
mfG for 4h and then subjected to live imaging
after washing with PBS. The fluorescence
signal Of mfG/PEG-FA/ICG was clearly
observed.
 Fluorescent image merged with DIC images
revealed that mfG Nano conjugates were
accumulated Into the cytoplasm .
 The release of doxorubicin by using confocal
fluorescence microscopy. We observe the
release of Dox after 4h from incubation .
 PBS – Phosphate-buffered saline used in
biological research. (PH ~ 7.4)

PHOTOTHERMAL PROPERTIES OF GRAPHENE :
 The heating of nanomaterials under NIR enhances the
activity of Dox at high temperature.
 Hela cells were incubated with BSA, ICG and mfG/PEG-
FA/ICG/Dox for 4h and then have been irradiated for 10 min
using a 785 nm NIR laser at 1W.
 The viability of the cells was measured at 0 and 24h of post-
irradiation .
 Significant reduction of viability observed after 24 h likely to
be attributed to the synergistic effect of heat induced by
laser irradiation and a fast Dox release from mfG.
 Result : mfG/PEG-FA/ICG/Dox has potential to be used for
multimodal therapy (chemo- and phototherapy).

BIOSAFETY OF mfG NANOSHEETS :
In vitro toxicity:
 Incubated Hela cells with a wide range of concentration of
mfG. Results revealed that mfG did not affect cell viability even
up to the Concentration of Approx. 150 microgram per ml
indicating it is safe for biomedical use .
In vivo toxicity:
 Examined by key blood parameters. Mice were intravenously
injected 100 microlitre of the starting mfG. The hematological
and biochemical parameters did not differ underscoring the
biosafety of mfG.

CONCLUSION :
 Designed and synthesized a multifunctional Graphene-based
platform for cancer therapy application.
 We demonstrate combined chemo- and photothermal properties of
the mfG as drug delivery carrier.
 Targeting ability of folic acid for Hela cells.
 The in vitro and in vivo toxicity of starting material.
 Good cancer killing activity shown by composite Functionalized
with doxorubicin.
 Internalization was confirmed by live microscopy exploiting ICG as
fluorophore.
 Combined affect of photo and chemotherapy after NIR irradiation.

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