DNA interaction analysis with automated biosensor of paraben derivative s-triazines

Article abstract of DOI:10.1016/j.molstruc.2020.128925

Among all heterocyclic compounds, the triazine scaffold has an important place due to its wide range of biological activities. Triazine has been preferred by many researchers as the target structure for the design and development of new drugs, as it is found in many biologically active molecules with promising biological potential such as anti-inflammatory, anti-mycobacterial, anti-viral, anti-cancer properties. In this study, a series of paraben substituted s-triazine compounds 1-5 were synthesized and their structures were determined by ¹H and ¹³C NMR spectroscopies as well as mass spectrometry. Also, structure of compound 5 was adjusted by X-ray crystallographic technique and crystallographic analysis revealed that 5 was crystallized in the trigonal space group R-3. According to X-ray analysis, compound 5 has very important intermolecular interaction such as nonclassical H-bond and pi-pi interaction. These interactions lead formation of supramolecular network. The thermal stabilities were investigated by thermogravimetric analysis. According to thermogravimetric analysis results, all of the synthesized compounds have high thermal stability. DNA interaction studies of the compounds 1-5 were performed with an automatic biosensor device that can measure the efficiency of DNA hybridization on the biochip surface. The biosensor results show that benzyl paraben substituted s-triazine compound 5, the most reactive compound at low concentration, was showed 61percent and 75percent activity reduction at concentrations 12.5 and 25 μM.

Full text of DOI:10.1016/j.molstruc.2020.128925

Journal Pre-proof
DNA Interaction Analysis with Automated Biosensor of Paraben
Derivative s-Triazines
¨
Elif S¸ enkuytu , Derya Davarcı , Zehra Olc¸er ,
¨
Gonu¨l Yenilmez C¸ iftc¸i
PII:
S0022-2860(20)31250-3
DOI:
Reference:
https://doi.org/10.1016/j.molstruc.2020.128925
MOLSTR 128925
To appear in:
Journal of Molecular Structure
Received date:
Revised date:
Accepted date:
29 March 2020
16 July 2020
16 July 2020
¨
¨
Please cite this article as: Elif S¸ enkuytu , Derya Davarcı , Zehra Olc¸er , Gonu¨l Yenilmez C¸ iftc¸i , DNA
Interaction Analysis with Automated Biosensor of Paraben Derivative s-Triazines, Journal of Molecular
Structure (2020), doi: https://doi.org/10.1016/j.molstruc.2020.128925
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Highlights
 A series of paraben substituted s-triazine compounds were synthesized and
characterized.
 DNA interaction studies of the compounds were performed with an automatic
biosensor device.
 The benzyl paraben substituted s-triazine compound 5 was showed 61% and
75% activity reduction at concentrations 12.5 and 25 µM.
1

DNA Interaction Analysis with Automated Biosensor of
Paraben Derivative s-Triazines
Elif Şenkuytu1,2*, Derya Davarcı2, Zehra Ölçer2 and Gönül Yenilmez Çiftçi2
1Department of Chemistry, Science of Faculty, Atatürk University, 25240, Erzurum, Turkey
2Department of Chemistry, Gebze Technical University, Gebze 41400, Kocaeli, Turkey
Corresponding author:
Dr. Elif Şenkuytu,
e-mail: elif.senkuytu@atauni.edu.tr ; Tel: +90 442 231 4441
2

ABSTRACT
Among all heterocyclic compounds, the triazine scaffold has an important place due to its
wide range of biological activities. Triazine has been preferred by many researchers as the
target structure for the design and development of new drugs, as it is found in many
biologically active molecules with promising biological potential such as anti-inflammatory,
anti-mycobacterial, anti-viral, anti-cancer properties. In this study, a series of paraben
substituted s-triazine compounds 1-5 were synthesized and their structures were determined
1
by H and 13C NMR spectroscopies as well as mass spectrometry. Also, structure of
compound 5 was adjusted by X-ray crystallographic technique and crystallographic analysis
revealed that 5 was crystallized in the trigonal space group R-3. According to X-ray analysis,
compound 5 has very important intermolecular interaction such as nonclassical H-bond and
pi-pi interaction. These interactions lead formation of supramolecular network. The thermal
stabilities were investigated by thermogravimetric analysis. According to thermogravimetric
analysis results, all of the synthesized compounds have high thermal stability. DNA
interaction studies of the compounds 1-5 were performed with an automatic biosensor device
that can measure the efficiency of DNA hybridization on the biochip surface. The biosensor
results show that benzyl paraben substituted s-triazine compound 5, the most reactive
compound at low concentration, was showed 61% and 75% activity reduction at
concentrations 12.5 and 25 µM.
3

Keywords: s-Triazine, Parabens, DNA Binding, Electrochemical Biosensor
Introduction
According to the World Health Organization (WHO), cancer disease, the second leading
cause of death worldwide, is responsible for an estimated 9.6 million deaths in 2018.
Globally, 1 out of 6 deaths (approximately 17% of all deaths) is due to cancer. Also, it is
known that approximately 70% of cancer deaths occur in low- and middle income countries
[1,2]. Parabens have been known for many years with their antimicrobial, antifungal
properties and are widely used in the cosmetic and pharmaceutical industry [3,4]. Also,
studies on the biological properties of parabens are frequently done [5,6]. In recent years,
researchers have been great effort to identify molecules with anticancer properties from both
natural and synthetic sources. Among the wide range of synthetic compounds known as
potential anticancer drugs, molecules based on triazine scaffolds drew great attention [7,8].
One of the triazine isomers, s-triazine (1,3,5-triazine), is a symmetrical and six-
membered nitrogen-containing heterocyclic compound. s-Triazine is a very useful scaffold for
multifunctional molecular arrangements and a stepwise substitution reaction because it
provides progressive substitution over three chlorine atoms [9-12]. s-Triazine has been used
as starting compound for preparing of wide variety derivatives bearing the s-triazine moiety,
due to the low-cost chemical, commercial availability, and easily substitution of the three
chlorine atoms with many of nucleophiles under controlled temperature [13-18]. In addition,
the chemical and physical properties of triazine derivatives can be modulated by the type,
number and orientation of functional side groups [19-24] and substituted triazines are very
stable, do not deteriorate under very aggressive chemical conditions. These fascinating
compounds derived from triazine have been examined intensively and been the subject of
many academic articles because of their widely applications in the pharmaceutical, cosmetic,
textile, plastic industries. Triazine derivatives are used as pesticides, dyestuffs, optical
4

bleaches, explosives, and surface-active agents. Triazine derivatives have been studied
especially in applications of medicinal chemistry [25-28]. The triazine scaffold provides the
source for the design of biologically related molecules in many studies such as antileishmanial
[29, 30], antimalarial [31], antimicrobial [32-34], antibacterial [35, 36], anticancer [37, 38]
and antitumor [39, 40].
DNA (deoxyribonucleic acid) provides chemists a very powerful tool to monitoring a
wide variety of diseases such as viral infections, infectious diseases. While molecules that
damage DNA can cause cancer, their role as anti-cancer drugs is very important.
Understanding of how molecules interact with DNA is important for the design of new drugs
[41-43]. There are a lot of studies in literature have used biosensors for DNA interaction
analysis and chemical toxicity tests [44, 45]. Electroanalytical techniques represent highly
sensitive tools for determination of chemical carcinogen compounds or mutagens and their
toxic effects on DNA [46, 47]. Although electroanalytical techniques have been proven to be
a useful tool for genotoxicity tests, lack of automated and fast devices prevented its use as the
routine toxicology analysis. Among the different detection methods, electrochemical DNA
biosensors have been used effectively to study the interactions of DNA with various
molecules [48, 49]. Moreover, due to the attractive advantages such as high sensitivity, low
cost and minimum power requirements, electrochemical DNA biosensors are rapidly
developing in clinical diagnosis and drug analysis areas. In this context, in the study, reactions
of s-triazine with methyl-, ethyl-, propyl-, butyl- and benzyl-paraben were carried out in
tetrahydrofuran, and fully paraben-substituted s-triazine compounds 1-5 were synthesized,
respectively. The structures of s-triazine series compounds 1-5 were characterized by
1
13
MALDI-TOF mass analysis, H and C NMR spectroscopies. Also, the solid-state structure
and geometry of compound 5 was determined using single-crystal X-ray structural analysis.
The thermal stability of compounds 1-5 were investigated by thermogravimetric analysis.
5

Biological activities and DNA interaction studies of the compounds 1-5 were also examined
by automatic biosensor device.
Scheme. Reactions of s-triazine with paraben derivatives
2. EXPERIMENTAL SECTION
2.1. General material and methods
1,3,5-s-triazine and paraben derivatives were obtained from Aldrich. Methyl 4-
hydroxybenzoate (99.0%), ethyl 4-hydroxybenzoate (99.0%), n-propyl 4-hydroxybenzoate
6

(99.0%), n-butyl 4-hydroxybenzoate (99.0%) were obtained from Alfa Aesar and benzyl 4-
hydroxybenzoate (99.0%) was obtained from Aldrich. Tetrahydrofuran (THF) (≥99.0%),
dichloromethane (DCM) (≥99.0%), n-hexane (≥95.0%) were bought from Merck. THF was
distilled over a sodium–potassium alloy under an atmosphere of dry argon. Silica gel 60
(230–400 mesh) for column chromatography was obtained from Merck. CDCl3 for NMR
spectroscopy was obtained from Goss Scientific. Positive ion and linear mode MALDI-MS of
compounds were obtained in dihydroxybenzoic acid as MALDI matrix using nitrogen laser
accumulating 50 laser shots using Bruker Microflex LT MALDI-TOF mass spectrometer. All
reagents were purchased from Aldrich and used without further purification and all solvents
were obtained from Merck. All reactions were monitored by thin layer chromatography using
Merck TLC Silica gel 60 F254. Silica gel 60 (particle size: 0.040-0.063 mm, 230-400 mesh
ASTM) for column chromatography was obtained from Merck. All reactions were carried out
1
under an argon atmosphere. H and 13C NMR spectra were recorded for all compounds in
CDCl3 solutions on a Varian INOVA 500 MHz spectrometer. Thermogravimetric analyses
(TGA) were performed on a Mettler Toledo TGA/SDTA 851. Powder samples of the
compounds were loaded into pans and heated at a ramp rate of 10 °C min−1 from room
temperature to 700 °C under a N2 atmosphere.
2.2. X-ray Crystallography
Suitable single crystals of compound 5 was carefully separated from other crystals in
the crystallization vessel under a polarizing microscope and placed on a thin glass fiber using
perfluoropolyether oil. The solid-state structures of the complexes were confirmed by X-ray
crystallography. Data were obtained on a Bruker APEX II QUAZAR three-circle
diffractometer using monochromatized Mo Kα X‒radiation (λ = 0.71073 Å). Indexing was
performed using APEX2 [50]. Data integration and reduction were carried out with SAINT
[51]. Absorption correction was performed by multi-scan method implemented in SADABS
7

[52]. The structure was solved using SHELXT [53] and then refined by full-matrix least-
squares refinements on F2 using the SHELXL [54] in OLEX 2 program package [55]. All
non-hydrogen atoms were refined anisotropically using all reflections with I > 2σ(I).
Aromatic and aliphatic C-bound H atoms were positioned geometrically and refined using a
riding mode. Crystallographic data and refinement details of the data collection for 5 were
given in Table S1. Crystal structure validations and geometrical calculations were performed
using Platon software [56]. Mercury software [57] was used for visualization of the cif files.
Additional crystallographic data with CCDC reference number (1964264 for 5) has been
deposited
within
the
Cambridge
Crystallographic
Data
Center
via
www.ccdc.cam.ac.uk/deposit.
2.3. Biochip, Biosensor device and DNA interaction tests
General information about biochip, biosensor device and DNA interaction tests were
given in Supporting Information.
2.4. Synthesis
Synthesis methods of the compounds are very similar. Therefore, the details of the
synthesis sections of the compounds were given in the supporting information.
Spectral data of compound 1: MS (MALDI-TOF) m/z (%): Calc. for (C27H21N3O9): 531.13;
found 531.97 [M]+ (Fig.S1). 1H NMR, CDCl3, 298 K: δ ppm, δH 7.86 (d, 3JHH = 8.75 Hz, 6H,
3
13
Ha); 7.00 (d, JHH = 8.75 Hz, 6H, Hb); 3.93 (s, 9H, -OCH3) (Fig.S2). C NMR (126 MHz,
CDCl3, 293 K, δ ppm): 173.29(C1), 166.02(C6), 154.80(C2), 131.27(C4), 128.24(C5),
121.38(C3), 52.26(C7) (Fig.S3).
Spectral data of compound 2: MS (MALDI-TOF) m/z (%): Calc. for (C30H27N3O9): 573.56;
1
3
found 5734.46 [M+H]+ (Fig.S4). H NMR, CDCl3, 298 K: δH ppm, 8.07 (d, JHH = 8.41 Hz,
8

3
6H, Ha); 7.21 (d, JHH = 8.41 Hz, 6H, Hb); 4.42-4.36 (m, 6H, -OCH2); 1.43-1.38 (m, 9H, -
CH3) (Fig.S5). 13C NMR (126 MHz, CDCl3, 293 K, δ ppm): 173.09(C1), 165.54(C6),
154.72(C2), 131.24(C4), 128.59(C5), 121.32(C3), 61.15(C7), 14.30(C8) (Fig.S6).
Spectral data of compound 3: MS (MALDI-TOF) m/z (%): Calc. for (C33H33N3O9): 615.64;
found 616.08 [M+H]+ (Fig.S7). 1H NMR, CDCl3, 298 K: δH ppm, 8.08 (d, 3JHH = 8.53 Hz, 6H,
3
Ha); 7.22 (d, JHH = 8.53 Hz, 6H, Hb); 4.41-4.27 (m, 6H, -OCH2); 1.84-1.76 (m, 6H, -CH2);
13
1.06-1.01 (m, 9H, -CH3) (Fig.S8). C NMR (126 MHz, CDCl3, 293 K, δ ppm): 173.30(C1),
165.60(C6), 154.71(C2), 131.25(C4), 128.62(C5), 121.33(C3), 66.73(C7), 22.09(C8), 10.48(C9)
(Fig.S9).
Spectral data of compound 4: MS (MALDI-TOF) m/z (%): Calc. for (C39H39N3O9): 657.72;
1
3
found 658.05 [M+H]+ (Fig.S10). H NMR, CDCl3, 298 K: δH ppm, 8.07 (d, JHH = 8.67 Hz,
3
6H, Ha); 7.22 (d, JHH = 8.67 Hz, 6H, Hb); 4.36-4.31 (m, 6H, -OCH2); 1.79-1.72 (m, 6H, -
CH2); 1.53-1.44 (m, 6H, -CH2); 1.02-0.97 (m, 9H, -CH3) (Fig.S11). 13C NMR (126 MHz,
CDCl3, 293 K, δ ppm): 173.30(C1), 165.59(C6), 154.70(C2), 131.26(C4), 128.64(C5),
121.33(C3), 65.03(C7), 30.74(C8), 19.25(C9), 13.74(C10), (Fig.S12).
Spectral data of compound 5: MS (MALDI-TOF) m/z (%): Calc. for (C45H33N3O9): 759.77;
found 759.06 [M]+ (Fig.S13). 1H NMR, CDCl3, 298 K: δH ppm, 8.12 (d, 3JHH = 8.69 Hz, 6H,
3
Ha); 8.75 (d, JHH = 6.70 Hz, 6H, -CH); 7.44-7.40 (m, 6H, -CH); 7.39-7.35 (m, 3H, -CH);
3
7.22 (d, JHH = 8.69 Hz, 6H, Hb); 5.37 (s, 6H, -OCH2), (Fig.S14). 13C NMR (126 MHz,
CDCl3, 293 K, δ ppm): 173.27(C1), 165.34(C6), 154.87(C2), 135.85(C8), 131.45(C4), [128.62,
128.31, 128.25, 128.16 (C5/C9/ C10/C11)], 121.42(C3), 66.90(C7) (Fig.S15).
3. RESULTS AND DISCUSSION
3.1. Synthesis and characterization of the compounds (1-5)
9

The synthesis of paraben-triazine compounds have been mentioned in a Patent [58]. The
reported procedure uses toluene as the solvent and pyridine as the proton abstractor. To avoid
the use of pyridine and also to replace high boiling toluene as solvent, we developed a new
methodology for the synthesis of paraben substituted triazine compounds. In the current
study, paraben substituted triazine compounds 1-5 were successfully synthesized and their
synthesis strategies were summarized in Scheme. The potential biological properties of the
parabens and paraben derivative compounds are known [59-62]. The triazine ring is very
useful platform for preparing of new biological systems due to its ability to combining
different functional groups. In these frameworks, reactions of 1,3,5-s-triazine with methyl-,
ethyl-, propyl-, butyl- and benzyl paraben were then performed in the presence of sodium
hydride and tetrahydrofuran to obtain full paraben substituted triazine compounds 1-5. All
the synthesized compounds were purified by column chromatography and each of the
1
obtained compounds 1-5 was characterized by mass (MALDI-TOF), H and 13C NMR
spectroscopies. The molecular structure of compound 5 was also determined by X-ray
crystallography. The appropriate crystallographic data was reported in Table S1. The structure
1
analysis data (mass, H and 13C NMR results) of the compounds are presented in the
experimental section. Also, the spectral data of compounds 1-5 are given in the supporting file
(Fig. S1-S31).
13
The 1H NMR chemical shift-coupling constants of all protons and C NMR chemical
shifts of all carbons in the experimental part confirmed the structures of synthesized
compounds 1-5. In general, the aromatic and aliphatic protons for paraben-substituted
triazines 1-5 were observed between 8.12-7.00 and 5.37-0.97 ppm, respectively. All integral
values for compounds 1-5 confirmed the proposed structures. Also, while aromatic carbons of
triazine compounds were observed between 173-121 ppm, aliphatic carbons were observed
13
1
13
among 66-10 ppm in C NMR spectra. The H and C NMR spectra of compound 4 was
10

depicted as an example in Fig 1a and 1b. While the -Ha and -Hb protons were observed as
double peaks at 8.07 and 8.67 ppm, the -OCH2 protons were seen as triple peaks at
approximately 4.33 ppm. in addition, the -CH2 and -CH3 protons were observed as multiple
peaks in the range of 1.79-1.44 ppm and as triple peaks at approximately 1.00 ppm,
respectively (Fig. 2a).
1
Figure 1. NMR spectra of the compound 4 in CDCl3 solution (a) H NMR spectrum and (b)
13C NMR spectrum.
3.2. Thermal Stabilities of Compounds 1-5
All of the compounds were stable in air at room temperature. The thermal stability of
the 1,3,5-s-triazine and paraben substituted triazine compounds (1-5) were investigated by
determining the weight loss of a sample upon linearly increasing the temperature by
°
°
conventional thermogravimetric analyses (TGA) at a heating rate of 10 C/min up to 700 C
11

under nitrogen flow. The thermal degradation curves of the compounds were given in Fig. 2.
The weight loss of 99.5 % was observed at 154.6 °C for 1,3,5-s-triazine. Synthesized
compounds 1- 5 have one-step thermal disruption curves. The compounds 1-5 were thermally
stable at 370.9 °C, 401.6 °C, 394.2 °C, 381.5 °C, 414.5 °C and remained amounts of
unspoiled were 22.1 %, 13.6 %, 7.7 %, 6.8 %, 17.6 %, respectively. The thermal stability of
1,3,5-s-triazine was lower than compounds 1-5. According to thermogravimetric analyses, all
of the synthesized compounds have high thermal stability and they are suitable for use in the
construction of high temperature resistant organic materials.
Figure 2. TGA curves of 1-5.
3.3. Crystal Structure of Compound 5
The X-ray structure of compound 5 was consistent with those proposed on the basis of
the NMR assignments. Crystallographic data, selected bond lengths and angles of compound
5 were summarized in Table S1-S3. Crystallographic analysis revealed that 5 was crystallized
in the trigonal space group R-3. Compound 5 consist of a six-membered ring N3C3, which is
fully substituted benzyl paraben on the C atoms (Fig. 3). As given in Table S2, the N1-C1i
and N1-C1 bond lengths are 1.333 (3) and 1.314 (3) Å; the N-C-N and C-N-C bond angles are
127.5 (2)°-112.5 (9)°, respectively. These values are typical for triazine derivatives [63-66].
Non-covalent interactions with H-bonds or aromatic clouds such as C-H…π, XH···π and π···π
play an important role for formation of stable supramolecular network. It was seen that there
12

were some short interactions for compound 5 molecule and these values were given in Table
S4a and 4b. As an example, 2D supramolecular structure like honeycomb (Fig. 4) formed
through the ab plane via short interaction of C9-H9…π(Cg3) (Fig. 5). In addition, the strong
aromatic interaction between the triazine rings along the c axis causes π···π stacking, which
leads to formation of the 3D supramolecular network. The distance between the N3C3 rings
(Cg1-Cg1) is 3.36 Å. (Fig 6).
Figure 3. ORTEP drawing of compound 5 (30% probability level) The grey, blue and red
coloured atoms represent C, N, O, respectively. All hydrogen atoms in 5 are omitted for
clarity. (Symmetry codes: (i) −x+y+1, −x+1, z; (ii) −y+1, x−y, z.).
13

Figure 4. C9-H9A…Cg3 interaction caused like honeycomb 2D supramolecular network
along the ab plane for compound 5
Figure 5. Perspective view of C9-H9…π (Cg3) intera2ction between compound 5 molecules
14

Figure 6. Perspective view of π···π stacking interaction (red dotted line) between N3C3 rings
(Cg1 rings) along the c axix.
Puckering analysis is a measure of ring conformation. It is calculation of the mean plane and
puckering parameters that afford a quantitative descriptor, the total puckering amplitude (Q)
[67]. In compound 5, N3C3 ring is almost planar (Fig. 7) and total puckering amplitudes for 6-
membered-ring couldn’t be analyzed (No C & P - Puckering Analysis since <Tau> = 0.1 <
5.0 Deg.) [68, 69]. QT values for N3C3 ring and deviations of nitrogen and carbon atoms from
the ring plane are given in Table S3.
Figure 7. The conformation of N3C3 ring for compound 5.
3.4. DNA Interaction Analysis with Automated Electrochemical Biosensor of the
Compounds 1-5.
DNA interaction assays with paraben substituted s-triazine compounds 1-5 were carried
out with MiSens® device, an automated electrochemical biosensor with sensitivity and fast
response [70-72]. The interactions consist of several steps on microfluidic channel integrated
15

biochip during flow (Fig. S16). It was studied a successful alkanethiol coating on the biochip
surface and the effect of the alkanethiol construction was measured using potassium
ferrocyanide solution. In Fig. S17 showed that electrochemical reduction and oxidation area
of ferrocyanide was larger on bare gold sensor chip than alkanethiol coated chip, and this
evidenced the formation of self-assembled monolayer for the NeutrAvidin immobilization.
For the investigation of synthesized compounds firstly, the biotinylated surface probe DNA
was captured on Neutravidin immobilized biochip surface. Hybridized complementary target
sequence and biotinylated detection probe DNA were incubated with varying concentrations
(0-50 μM) of compounds and then this mixture was injected to the capture probe immobilized
biochip surface in order to achieve the hybridization process of complementary surface probe
(capture probe). In the next step, Neutravidin and HRP enzyme immobilized gold
nanoparticles were injected to the biochip surface for binding via neutravidin and biotin
interaction, and then an amperometric biosensor signal was measured during the substrate
TMB injection (Fig. S18).
The individual reactive parabens (methyl-, ethyl-, propyl-, butyl-, and benzyl-
parabens), 1,3,5-s-triazine and the paraben substituted s-triazine compounds 1–5 were tested
by biosensor device and the hybridization activity results are shown Fig. 8 and Fig. S19
indicated by remaining from interaction and interacting DNA. While the individual parabens
interact with DNA at a rate of 35-20%, it was observed that the substituted-s-triazine
compounds 1-5 to which these parabens bind interact with DNA in approximately 70% at 50
μM concentration. Especially, when the hybridization activity of the synthesized products has
been compared to parabens, they have shown the high activity reduction in low concentrations
(12.5 and 25 μM) (Fig. 8 and Fig. S19). It was observed that among the synthesized
compounds, benzyl-paraben substituted triazine (5) was the most reactive compound at low
concentration. Compound 5 has shown 61% and 75% activity reduction in 12.5 and 25 μM
16

concentrations, respectively. At low concentration, 12.5 μM, compounds 2, 3, and 4 have also
shown similar good activity reduction, approximately 65% DNA interaction.
(a)
100
100
100
100
100
92
100
80
60
40
20
0
65
57
52
47
39
48
45
44
42
34
0
25
36
32
12.5
23
25
50
1
2
3
4
5
Compounds
100
90
80
70
60
50
40
30
20
10
0
(b)
Comp. 1
Comp. 2
Comp. 3
Comp. 4
Comp. 5
0
10
20
30
40
50
Concentration (µM)
Figure 8. The amperometric responses were shown as percent relative hybridization
responses in the figures, indicated by remaining from interaction (a) and interacting DNA (b).
Compounds (0, 12.5, 25 and 50 µM) were incubated with DNA probes prior to the injection
on to the biochip.
17

4. CONCLUSION
In summary, in this study, a series of paraben substituted s-triazine compounds were
successfully synthesized and DNA interactions of these compounds 1-5 were investigated by
an automated biosensor device. The structural properties of all synthesized compounds were
1
examined by MALDI-TOF spectrometer, H, 13C NMR spectroscopy. The solid-state
structure and geometry of compound 5 was determined using single crystal X-ray structural
analysis. In addition, it was confirmed by thermogravimetric analyses (TGA) that all
compounds 1-5 showed high thermal stability. Generally, while the parabens interact with
DNA at a rate of 35-20%, it was observed that the paraben substituted-s-triazine compounds
interact with DNA in approximately 70% at 50 μM concentration. Especially, benzyl-paraben
substituted s-triazine compound 5, which the most reactive compound at low concentration,
was showed 61% and 75% activity reduction in 12.5 and 25 μM concentrations, respectively.
Compound 5 is highly effective in DNA binding activity so, this compound may be a
potential candidate as anticancer agents. It is important to quickly determine the effects of
drug candidate molecules by pre-screening tests. In this context, data from biosensor analysis
studies can help researchers select the most likely drug candidates for further research. In the
future application of the study, it is aimed to investigate the anticancer capabilities of the
compounds against different cancer cells.
AUTHOR INFORMATION
*Corresponding author: Dr. Elif Şenkuytu
e-mail: elif.senkuytu@atauni.edu.tr , Tel: +90 442 231 4441
SUPPLEMENTARY MATERIAL
Supplementary data were given as Supporting Information. Structure determination has been
deposited with the Cambridge Crystallographic Data Centre with references CCDC 1964264
18

for compound 5. These data can be obtained free of charge from The Cambridge
Crystallographic Data Centre via www.ccdc.cam.ac.uk/data_request/cif.
Sample CRediT author statement
Elif Şenkuytu: Formal analysis, Writing- Original draft preparation, Supervision.
Derya Davarcı: Investigation and Writing
Zehra Ölçer: Investigation and Writing
Gönül yenilmez Çiftçi: Reviewing and Editing
Declaration of interests
☒ The authors declare that they have no known competing financial interests or personal
relationships that could have appeared to influence the work reported in this paper.
ACKNOWLEDGEMENTS
This work was supported by grants from the Gebze Technical University Scientific Research
Projects (GTUBAP) (Grant no: BAP 2018-A105-34). We would like to thank BILGEM-
TUBITAK for their contribution to the manufacturing of biochip and detection platform.
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