Synthesis, photophysical properties and DNA-Binding Studies of p-Hydroxy and m-Hydroxy Amphiphilic Porphyrins

Article abstract of DOI:10.14233/ajchem.2014.16441

Three p-hydroxy and m-hydroxy amphiphilic porphyrins had been synthesized and fully characterized by 1H NMR, IR and FAB-MS. Their photophysical properties were also determined. Furthermore, the UV-visible absorption and fluorescence emission titration exp

Full text of DOI:10.14233/ajchem.2014.16441

Asian Journal of Chemistry; Vol. 26, No. 3 (2014), 933-936
http://dx.doi.org/10.14233/ajchem.2014.16441
Synthesis, Photophysical Properties and DNA-Binding Studies of
p-Hydroxy and m-Hydroxy Amphiphilic Porphyrins
*
JIE ZHANG , HAIWEI JIN, DONGCHUN GAO, WENPING JIA, FANG LI, PING ZHANG, TONGTONG SONG and JIALI WANG
School of Pharmaceutical and Chemical Engineering, Taizhou University, Taizhou 318000, P.R. China
*
Corresponding author: Tel: +86 57688660177; E-mail:zhang_jie@tzc.edu.cn
Received: 28 September 2013; Accepted: 12 November 2013;
Published online: 30 January 2014;
AJC-14671
1
Three p-hydroxy and m-hydroxy amphiphilic porphyrins had been synthesized and fully characterized by H NMR, IR and FAB-MS.
Their photophysical properties were also determined. Furthermore, the UV-visible absorption and fluorescence emission titration experiments
indicate that porphyrins and DNA existed an outside binding mode. In addition, the DNA binding constant of 4pOH-TPP, 3pOH-TPP and
4mOH-TPP were determined.
Keywords: Porphyrins, Amphiphilic, Photophysical, DNA-binding.
INTRODUCTION
EXPERIMENTAL
During the past two decades, a great interest has been
The chemicals were purchased from Sigma-Aldrich Com-
pany and used as received. Silica gel 60 (0.04-0.063 mm) for
column chromatography was purchased from Merck. NMR
spectra were recorded on a BrukerAvance III 400MHz NMR
drawn to porphyrin-based materials. The porphyrins are known
for their multiple biological functions and excellent metal
complexing ability. And, porphyrin-based materials were
1
1
widely used in many areas including photodynamic therapy ,
spectrometer. Chemical shifts of H NMR spectra were refer-
enced to internal deuteriated solvents and then recalculated to
2
3
4
antiviral , anticancer therapies , NLO materials , OLED and
5
solar cell . Porphyrins and its derivatives are also one of
6
themost studied DNA binding agents . According to some
4
SiMe (δ0.00). FAB mass spectra were obtained with a Quattro
micro API mass spectrometer. Electronic absorption spectra
in the UV-Visible region were recorded with a Shimadzu UV-
2401 PC spectrophotometer instrument. The IR spectra (KBr
pellets) were recorded with a Nicolet 5700 FTIR spectrometer.
Steady-state visible fluorescence and PL excitation spectra
were recorded on a Cary Eclipse spectrofluorimeter.
previous studies, there are several binding modes to native
and synthetic double stranded DNA. The binding modes were
marked affected by periphery groups, positive charges and
the nature of DNA of the porphyrin derivatvies. It has been
shown that the grooving binding, outside random binding
and stacking along the DNA template are the common bonding
General procedure
7
modes .
It was reported that hydroxylporphyrins derivatives
8
showed good water solubility and bioactivities . Moreover,
3p-OH-TPP: A solution of benzaldehyde (0.53 g, 5 mmol)
and p-hydroxybenzaldehyde (1.83 g, 15 mmol) in propionic
acid (200 mL) was heated to 130 °C. Then freshly distilled
pyrrole (1.34 g, 20 mmol) in propionic acid (50 mL) was added
slowly to the solution over a period of 0.5 h. The reaction
mixture was heated to reflux for another 0.5 h and then cooled
to room temperature. Then the reaction mixture was reduced
to dry under reduced pressure. The resultant was chromato-
these porphyrins could be easily synthesized and separated.
However, to the best of our knowledge, there are few reported
about the DNA bonding properties of the hydroxylporphyrins.
For such reasons, we prepared a series of three p-hydroxy and
m-hydroxy amphiphilic porphyrins, in which a hydroxy is
attached to the para and meta position. The main purpose of
this paper is to reveal the DNA binding mode of the hydroxyl
amphiphilic porphyrins and CT-DNA. At the same time, the
graphed on a silica gel column with CHCl
band gave the desired 3p-OH-TPP.Yield: 106 mg (3.2 %). H
NMR (CDCl ): δ = 8.86-8.90 (m, 8H), 8.23-8.25 (m, 4H),
8.09-8.11 (m, 4H), 7.76-7.81 (m, 5H), 7.23-7.25 (m,4H) and
3
as eluent. The fourth
1
1
novel porphyrins were fully characterized by IR, MS and H
3
NMR.

934 Zhang et al.
Asian J. Chem.
-
2.75 (s, 2H). FAB MS (positive mode): m/z = 647 [M-OH +
acid to give the product 4m-OH-TPP (4.4 %). The amphiphilic
porphyrins showed good solubility in common organic solvents
+
-1
H] . IR (KBr, νmax, cm ): λ = 3508, 1608, 1510, 1350, 1267,
171, 966, 799 and 723.
p-OH-TPP: benzaldehyde (0.53 g, 5 mmol), p-hydroxy-
1
3 2 2 3
like CHCl , CH Cl , CH OH, DMF and DMSO etc. The comp-
1
lexes were fully characterized by H NMR, IR, FAB-MS, UV-
4
1
visible spectrum and fluorescence analysis. The H NMR spectra
benzaldehyde (1.83 g, 15 mmol) and propionic acid (200 mL)
were used. The fifth band gave the desired 4p-OH-TPP.Yield:
of hydroxyl amphiphilic porphyrins show singlets at δ = -2.75
to -2.96 ppm for the inner NH protons of the porphyrin rings
(experimental section).
1
00 mg (5.9 %). H NMR (CDCl
2
8
7
6
1
3
): δ = 8.86-8.90 (m, 8H),
.22-8.25 (m, 4H), 8.09-9.11 (m, 4H), 7.77-7.79 (m, 4H), 7.24-
.26 (m, 4H) and -2.75 (s, 2H). FAB MS (positivemode): m/z =
Absorption titrations: The UV-visible absorption spectra
of amphiphilic porphyrinsin the absence and presence of
increasing amounts of CT DNA are given in Fig. 1. The UV-
visible absorptions of these amphiphilic porphyrins retain the
+
47 [M-2OH + H] . IR (KBr, νmax, cm ): λ = 3945, 1599, 1506,
-1
469, 1340, 1261, 1171, 970, 800 and 711.
4
m-OH-TPP: A solution of m-hydroxybenzaldehyde
2.44 g, 20 mmol) in propionic acid (200 mL) was heated to
30 °C. Then freshly distilled pyrrole (1.34 g, 20 mmol) in
(
2
characteristics features of H TPP. They show a strong Soret
1
band in the blue region and weak Q-bands in the visible region.
The titration of DNA induces large spectral perturbations in
the Soret bands of all the porphyrins. The hypochromism reach
58.0, 61.4 and 42.8 % for 4pOH-TPP, 3pOH-TPP and 4mOH-
TPP, respectively (Table-1). These porphyrins exhibited a
remarkable decrease in intensity, but the absorption bands do
not have any red shift with the titration of DNA.
propionic acid (50 mL) was added slowly to the solution over
a period of 0.5 h. The reaction mixture was heated to reflux
for another 0.5 h and then cooled to room temperature. Then
the reaction mixture was reduced to dry under reduced pres-
sure. The resultant was chromatographed on a silica gel column
1
as eluent.Yield: 149 mg (4.4 %). H NMR (CDCl
with CHCl
3
3
):
δ = 9.90 (s, 4H), 8.90 (m, 6H), 7.61-7.63 (m, J = 8.4Hz, 10H),
.32-7.39 (m, 2H), 6.85-7.08 (m, 2H)and -2.96 (s, 2H). FAB
The UV-visible spectra of amphiphilic porphyrins show
no red shift and hypochromism of Soret bands from 42.8 to
61.4 %, which indicated the porphyrins adopt an outside
7
MS (positive mode): m/z = 679 [M] . IR (KBr, νmax, cm ): λ =
+
-1
9
3
8
387, 1701, 1582, 1440, 1288, 1223, 1163, 1080, 922 and
02.
binding mode .All physical data about UV-titration experiments
are listed inTable-1.
Emission titrations: The fluorescence spectra of these
amphiphilic porphyrins also retain the characteristics features
RESULTS AND DISCUSSION
Synthesis of hydroxy amphiphilic porphyrins: The
2
of H TPP. The products emitted at about 650 and 720 nm. The
synthetic routes for the preparation of hydroxy amphiphilic-
porphyrins are shown in Scheme-I. p-hydroxybenzaldehyde,
benzaldehyde and pyrrole was refluxed in propionic acid to
give the products 3p-OHTPP (3.2 %) and 4p-OHTPP (5.9 %).
m-Hydroxybenzaldehyde and pyrrole was refluxed in propionic
fluorescence spectra and data of amphiphilic porphyrins are
shown in Fig. 2. We also carried out the emission titration
experiments for 4pOH-TPP, 3pOH-TPP and 4mOH-TPP with
DNA (Fig. 2). In the presence of CT DNA, a decrease in intensity
of fluorescence emission is depicted for all porphyrins (21.4-
OH
OH
CHO
CHO
propionic acid
NH
N
NH
N
+
+
OH
+
HO
HO
N
H
N
HN
reflux
N
HN
OH
OH
p-OH-TPP
OH
OH
p-OH-TPP
3
4
CHO
CHO
HO
propionic acid
reflux
NH
N
N
+
+
HO
N
H
HN
OH
HO
4m-OH-TPP
Scheme-I: Synthesis of 4p-OHTPP, 3p-OHTPP and 4m-OHTPP

Vol. 26, No. 3 (2014)
Synthesis, Properties and DNA-Binding Studies of Amphiphilic Porphyrins 935
TABLE-1
CHANGES IN ELECTRONIC SPECTRA OF AMPHIPHILIC PORPHYRINS IN THE
ABSENCE OR PRESENCE OF CT-DNA IN tris-HCl BUFFER (pH = 7.2, 0.05 M NaCl)
UV–visible on the Soret band
Fluorescence emission
-
Kapp(M )
1
Compound
λmax (free) nm
λmax (bound) nm
∆λ (nm)
H (%)
Decrease of intensity (%)
7
-1
-1
-1
4
pOH-TPP
pOH-TPP
430
433
426
430
433
426
0
0
0
58.0
61.4
42.8
34.7
50.1
21.4
1.54 × 10 M
7
3
1.59 × 10 M
7
4
mOH-TPP
1.47 × 10 M
1
0
0
0
.0
.8
.6
.4
0.8
0.6
0
.5
.4
.3
.2
.1
.0
(
a)
(b)
(c)
0
0
0
0
0
0
.4
.2
0
0.2
0.0
0.0
400
500
600
700
400
500
Wavelength (nm)
600
700
400
500
600
700
Wavelength(nm)
Wavelength (nm)
Fig. 1. Absorption spectra of 4p-OHTPP (a), 3p-OHTPP (b) and 4m-OHTPP (c) in tris buffer (pH = 7.2, 0.05 M NaCl) at 25 °C in the presence of increasing
amounts of CT DNA. [Por] = 10 µM. Arrows indicate the change in absorbance upon increasing the DNA concentration
4
3
2
1
00
00
00
00
0
400
(
a)
(b)
(c)
3
2
1
00
00
00
0
300
200
100
6
00
650
700
750
600
700
6
00
700
Wavelength (nm)
Wavelength (nm)
Wavelength (nm)
Fig. 2. Emission spectra of 4p-OHTPP (a), 3p-OHTPP (b) and 4m-OHTPP (c) in trisbuffer (pH = 7.2, 0.05 MNaCl) at 25 °C in the presence of increasing
amounts of CT DNA. [Por] = 10 µM. Arrows indicate the change in emission upon increasing the DNA concentration
5
0.1 %, Table-1). This phenomenon indicated that self stack-
3.5
3.0
2.5
4pOH-TPP
3p-OHTPP
4m-OHTPP
ing of the neighbouring porphyrin molecules to each other
along the DNA surface.
DNA binding constant experiments: We carried out the
EB competitive binding experiments and the florescence
quenching plot are given in Fig. 3. The I
plot is good agreement with the linear Stern-Volmer equation
I/I = 1 + KQ) with a slope of 11.72, 12.02 and 10.65 for
pOH-TPP, 3pOH-TPP and 4mOH-TPP, respectively. We can
0
/I vs. [Por]/[DNA]
2.0
.5
1
(
0
4
1.0
0
also learn Fig. 3 that 50 % of EB molecules were replaced
from DNA-bound EB at a concentration ratio of [Por]/[EB] =
.00
0.05
0.10
0.15
0.20
R=[Por]/[DNA]
0
TPP, respectively. The Kapp of EB in the experimental condition
.65, 0.63 and 0.68 for 4pOH-TPP, 3pOH-TPP and 4mOH-
Fig. 3. Florescence quenching curves of DNA-bound EB by amphiphilic
porphyrins in tris buffer (pH = 7.2, 0.05 M NaCl). ([DNA] = 100
µM, [EB] = 16 µM, λexc = 537 nm)
7
is 1 × 10 M , therefore, the Kapp of 4pOH-TPP, 3pOH-TPP
-1
7
and 4mOH-TPP were 1.54 × 10 M and 1.59 × 10 M and
-1
7
-1
7
-1
10
.47 × 10 M , respectively (Table-1) .
to 61.4 %, which indicated the porphyrins adopt an outside
binding mode. The emission titration experiments indicate that
self stacking of the neighbouring porphyrin molecules to each
other along the DNA surface.The DNA binding constant
app
1
Conclusion
Three hydroxy amphiphilic porphyrins were prepared and
1
fully characterized by H NMR, IR, FAB-MS and UV-visible
experiments revealed the K of 4pOH-TPP, 3pOH-TPP and
7
4mOH-TPP were 1.54 × 10 M and 1.59 × 10 M and 1.47
-1
7
-1
spectroscopy. The UV-visible spectra of amphiphilic porphyrins
show no red shift and hypochromism of Soret bands from 42.8
7
-1
× 10 M , respectively.

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