Page 51 - TD-1-1

Tumor Discovery Dual-targeting of cancer receptors by nanoparticles

Ideally, all NDDSs should improve stability, absorption, High pure RNA isolation kit was obtained from Roche
drug concentration, and long-term release into the target (Rotkreuz, Switzerland). EasyTM cDNA synthesis kit was
tissue . Besides, other factors, such as reducing drug purchased from Pars Tous Biotechnology (Mashhad, Iran).
[4]
injections to increase patient comfort and advanced drug The dialysis bag (MWCO = 10 kDa) was purchased from
delivery systems, should be considered . NDDSs based Sigma-Aldrich (St. Louis, MO, USA).
[4]
on magnetic nanoparticle carriers (MNCs) have greatly
revolutionized the field of treatment and diagnosis , 2.2. Synthesis of Fe O -DPN-HA
[5]
3
4
magnetic resonance imaging, and hyperthermia . According to our previous work, Fe O nanoparticles
[6]
[7]
4
3
MNCs can also be used in targeted drug delivery (NPs) were synthesized by heating the mixture of
with regard to features, such as biocompatibility, iron(III) acetylacetonate in the presence of benzyl ether
[18]
superparamagneticity, stability , remotely adjustable and oleylamine at 120 and 270°C . Furthermore, the
[8]
capability, and easy synthesis . Nevertheless, the anchoring of DPN to Fe O was carried out at 25°C under
[10]
[9]
3
4
[13]
important issues with MNCs are their hydrophobic surface, an argon blanket .
which leads to a shorter presence in the bloodstream due To prepare Fe O -DPN-HA, it is necessary to activate
3
4
to opsonization through plasma protein . A proposed the carboxylic acid (−COOH) groups of HA. For this
[11]
method to overcome this disadvantage is coating MNCs’ purpose, in the first step, 44 mg of HA (0.11 mmol, equal
surfaces with different functional groups to reduce the to the number of moles of carboxyl groups) was dissolved
amount of opsonization and subsequently increase the in 5 mL formamide and stirred at 70°C for 5 h. After
circulation time of the particles in the bloodstream . cooling for some time to room temperature, 45.5 mg or
[12]
Hyaluronic acid (HA), a suitable polymeric ligand 0.22 mmol dicyclohexylcarbodiimide (DCC) and 25.4 mg
with components consisting of acetyl glucosamine and or 0.22 mmol N-Hydroxysuccinimide (NHS) were added
glucuronic acid, was applied for modification of MNCs. It to the mixture and stirred at room temperature for 24 h.
displays unique properties, such as biocompatibility, non- For immobilizing of activated HA on Fe O -DPN, 353 mg
3
4
toxicity, and the ability to interact with a variety of drugs of Fe O -DPN (containing 0.11 mmol of NH groups)
4
3
2
directly or through drug carriers [13-15] . was dispersed to the reaction mixture and stirred for 24 h
Cluster of differentiation-44 (CD44), a non-kinase at 40°C. The pH of the reaction mixture shifts into the
transmembrane glycoprotein and a major receptor for alkaline range using 200 µL of triethylamine. The products
HA, is overexpressed in certain types of cancer cells. (compound 2 shown in Figure 1) were separated by an
HA has gained much attention in cancer-targeting drug external magnetic field (DynaMag TM-50 system) and
delivery and could reduce drugs side effects because it can washed several times with acetone, and then, freeze-drying
[21]
attach to the CD44 receptor and enters the host cell by was performed at −60°C for 4 h (yield was 58.61%) .
endocytosis [13,16] .
2.3. Synthesis of tosylated-Fe O -DPN-HA (HA-OTs)
3
4
In addition to CD44, folate receptor protein, where folic
acid (FA) has a high affinity to it, has been overexpressed on A 50 mg of Fe O -DPN-HA (containing 0.024 mmol
3
4
of hydroxyl groups of HA) was dispersed in 5 mL of an
many malignant cancer cells. [17-19] . Therefore, in this study, alkaline buffer (pH 9.5) and stirred and cooled to 4°C in
we synthesized a TMX-nanocarrier in which HA and FA an ice water bath under an argon blanket for 5 min. Then,
groups were conjugated simultaneously as the targeting 5.94 mg (0.031 mmol) of p-toluenesulfonyl chloride (TsCl)
agent on the surface of Fe O -DPA-PEG-NH (Fe O - was added to the reaction and stirred for 24 h at 4°C.
3
4
4
2
3
DPN) to treat MDA-MB-231 breast cancer cells. Besides Subsequently, 5 mL of dichloromethane was added to the
[20]
the synthesis and characterization of the nanostructure, reaction mixture to remove any unreacted TsCl from the
drug loading, cytotoxicity, and apoptosis ability were also bottom of the reaction balloon. Then, acetone was added
determined using MTT assay and real-time polymerase and the solid products were separated with the DynaMag
chain reaction (RT-qPCR).
TM-50 system. Eventually, the precipitates were washed
2. Materials and methods twice with acetone and dried using freeze-drying (yield
was 74.02%).
2.1. Materials
All chemicals were purchased from Merck (Darmstadt, 2.4. Synthesis of Fe O -DPN-HA-NH 2
3
4
Germany). MDA-MB-231 breast cancer cell lines were Briefly, all products of the previous steps were dispersed
obtained from Pasture Institute (Tehran, Iran). All media to 7 mL of ethylenediamine (EDA) and stirred overnight
and cell culture components were obtained from either under an argon atmosphere at 50°C. The products
Caisson labs (North Logan, UT) or Gibco (Carlsbad, CA). (compound 4 shown in Figure 1) were washed with
Volume 1 Issue 1 (2022) 2 https://doi.org/10.36922/td.v1i1.41