Convenient synthesis of novel unmetalled and metallophthalocyanines bearing coumarin derivatives: synthesis, characterization, aggregation behaviors and antimicrobial activity

Article abstract of DOI:10.1007/s10847-016-0655-7

Metal-free and metallophthalocyanines 3–10 carrying four coumarins group on the periphery were prepared by cyclotetramerisation of a new precursor, namely 4-(4-Methyl-2-oxo-2H-chromen-7-yloxy)-phthalonitrile 1 and 4-(2-Oxo-2H-chromen-7-yloxy)-phthalonitri

Full text of DOI:10.1007/s10847-016-0655-7

J Incl Phenom Macrocycl Chem
DOI 10.1007/s10847-016-0655-7
ORIGINAL ARTICLE
Convenient synthesis of novel unmetalled
and metallophthalocyanines bearing coumarin derivatives:
synthesis, characterization, aggregation behaviors
and antimicrobial activity
R. Medyouni1 A. C. Mtibaa2 L. Mellouli2 A. Romerosa4 N. Hamdi1,3
•
•
•
•
Received: 25 June 2016 / Accepted: 1 September 2016
Ó Springer Science+Business Media Dordrecht 2016
Abstract Metal-free and metallophthalocyanines 3–10
carrying four coumarins group on the periphery were pre-
pared by cyclotetramerisation of a new precursor, namely
Introduction
Phthalocyanines have been the subject of a great deal of wide-
ranging research for over 60 years. Phthalocyanines properties
have interesting potential technical applications such as molec-
ular electronics, semiconductor and electrochromic display
devices, photovoltaic and solar cells, gas sensors, synthetic
metals, liquid crystals, optical disks, photodynamic therapy of
cancer, electrophotography, and non-linear optics [1–10].
On the other hand coumarins, comprise a very large and
important family of compounds that occur widely in nature.
They are found in a wide range of plants such as tonka
bean, vanilla grass and cinnamon. A number of coumarin
derivatives are used in the pharmaceutical industry as
precursor molecules for the synthesis of many synthetic
pharmaceutical compounds including anticoagulants and
vitamin K antagonists, while others are used in the treat-
ment of lymphedema [11]. Coumarin derivatives exhibit
antibacterial, antifungal, anticancer, anticoagulant, anti-
inflammatory [11–16]. The synthesis and characterization
of Pc compounds including four peripheral 6-hydroxy-4-
methylcoumarin substituents units on the periphery have
been studied previously. [17].
4-(4-Methyl-2-oxo-2H-chromen-7-yloxy)-phthalonitrile
1
and 4-(2-Oxo-2H-chromen-7-yloxy)-phthalonitrile 2 in the
presence of the corresponding divalent metal salts (Zn (II),
Cu(II) and Co(II). The aggregation behaviors of the
unmetalled 3 and metallophthalocyanines 5 were investi-
gated. The novel phthalonitrile derivatives 1–2 were syn-
thesized by the reaction between 4-nitrophthalonitrile with
7-hydroxy-4-methylcoumarin and 7-hydroxycoumarin respec-
tively in polar aprotic solvents (DMF or DMSO) in the pres-
ence of dry K2CO3 as base catalyst. The characterization of the
new products involved a combination of methods including
elemental analyses, IR, UV–vis, 1H NMR, 13C NMR and mass
spectroscopies. Most of the new compounds 3–10 exhibited
important antimicrobial activity.
Keywords Metallophthalocyanine Á Zinc Á Synthesis Á
Antimicrobial activity
In this regard, we report herein the synthesis of metal-
free phthalocyanines (3,4) and metallophthalocyanines
(5–10) carrying coumarins groups. The spectroscopic char-
acterization and the electronic and aggregation behaviors of
these newly synthesized complexes are also presented.
& N. Hamdi
naceur.hamdi@isste.rnu.tn
1
Chemistry Department, College of Science and Arts, Qassim
University, Al-Rass, Kingdom of Saudi Arabia
2
Laboratory of Microorganisms and Biomolecules of the
Center of Biotechnology of Sfax-Tunisia, Road of Sidi
Mansour, Km 6, B.P. 1117, 3018 Sfax, Tunisia
Results and discussion
3
Heterocyclic and Organometallic Chemistry Laboratory,
Higher Institute of Environmental Sciences and Technology,
University of Carthage, Hammam-Lif 2050, Tunisia
The synthesis of substituted phthalonitrile derivatives is an
important step in Pc synthesis. Nonperipherally substituted
phthalonitrile derivatives are synthesized through reactions
4
´
Area de Quımica Inorganica, Universidad de Almerıa,
´
04120 Almerıa, Spain
´
´
´
123

J Incl Phenom Macrocycl Chem
Scheme 1 protocol route for
the synthesis of compounds 1–2
R
O
R
O
HO
O
CN
O2N
CN
i or ii
CN
O
CN
O
4-nitrophthalonitrile
R = CH3 (1)
R = H (2)
R = CH3 ; (i) : K2CO3, dry DMF, r.t , 24h
R = H ; (ii) : K2CO3, DMSO, r.t , 72h
between 4-nitrophthalonitrile and O-, S-, or N-nucleophiles
[18–23]. Using this synthetic strategy, the synthesis and
characterization of metallophthalocyanines 5-10 and their
precursors 1–2 are reported. The 4-(4-Methyl-2-oxo-2H-
chromen-7-yloxy)-phthalonitrile 1 was obtained from the
reaction of 7-Hydroxy-4-methyl coumarin to 4-nitropph-
thalonitrile in the presence of K2CO3/DMSO under N2
atmosphere at room temperature for 24 h. The synthesis of
4-(2-oxo-2H-chromen-7-yloxy)-phthalonitrile 2 was achieved
in 74 % yield through base-catalyzed aromatic displace-
ment of 4-nitrophthalonitrile with 7-hydroxycoumarin
using K2CO3 as the base in dry DMF. The reaction was
carried out at room temperature under N2 atmosphere for
72 h (Scheme 1).
The characterization of the products 1–2 involved a
1
combination of methods including, H NMR, 13C NMR,
and NMR 2D, FT-IR, UV–vis, elemental analysis and LC–
MS spectroscopy.
In the IR spectrum of compound 1, stretching vibrations of
CN groups at 2227 cm-1 and aromatic groups (ArCH) at
1
3079 cm-1 appeared at expected frequencies. The H NMR
spectrum of 1 in DMSO-d6 showed signals with ranging from
7.43 to 8.11 ppm belonging to aromatic protons and H2,3. The
methyl protons appeared at 2.42 ppm. In the 13C NMR spectra
of 1, the aromatic, and CH3 carbon atoms of 1 appeared
between 116.2 and 161.7 ppm and at18.5 ppm, respectively,
whereas carbon atom C2 was observed at 159.8 ppm.
1H NMR spectral assignments are also supported via
1
The electron-withdrawing capability of dinitrile func-
tionalities makes 4-nitrophthalonitrile susceptible to
nucleophilic attack [24, 25].
1H–13C HMBC experiment. H–13C HMBC is more help-
ful to get detailed information about which protons corre-
late with certain carbons. From the 1H–13C HMBC spectra,
Scheme 2 Synthesis route for
unmetalled phthalocyanine
compound (3,4)
R
R
O
DMAE, 150°C, 24h
1
O
O
O
O
DBU
O
N
HN
N
N
N
N
NH
N
O
O
O
O
O
O
DMAE, 150°C, 24h
R
R
R = CH (3) ; R = H (4)
3
DBU
2
123

J Incl Phenom Macrocycl Chem
H3 proton at 7.43 ppm correlates with C2 at 159.8 ppm and
C4 at 153.2 ppm. The mass spectra of phthalonitrile
derivatives 1–2 gave the characteristic molecular ion peaks
at m/z = 287.2, 301.2 respectively.
phthalonitrile compound 1 suggested the formation of
compound phthalocyanine derivative 3.
1
The H NMR spectrum of compound 3 exhibited char-
acteristic signal for methyl (-CH3) at d: 2.42, the aromatic
protons appeared between 8.32 and 8.42 ppm, the typical
shielding of inner core protons could not be observed due
to the probable strong aggregation (especially for our
molecules, due to multiplicities of the numbers of aromatic
benzenes) of the molecules [26]. The elemental analysis
results confirmed the structure of the desired compound 3.
The sharp peak for the (C:N) vibrations of phthalonitriles 2
at 2225 cm-1 disappeared after conversion into metal-free 4.
Researchers generally use substituted phthalonitriles or
1,3-diimino-1H-isoindoles as starting materials for syn-
thesis of Pcs [27–29]. In this work, phthalonitrile com-
pounds (1 and 2) were also used as starting compounds for
synthesis of the metallo-Pcs (Zn, Cu, Co) due to mild
reaction conditions.
Comparison of the IR spectral data clearly indicated the
formation of phthlonitrile derivative 2 by the disappearance
of the phenolic OH of coumarins derivatives at 3448 cm-1
as expected and NO2 band of 4-phtalonitrile at 1538–1355
cm-1 and appearance intense absorption band at 2225
cm-1 for (2) corresponding to the CN stretching. The
characteristic carbonyl vibrations of coumarin lactone ring
were observed at 1731 cm-1.
Metal-free phthalocyanine compound (3) was synthesized
by heating compound 1 in 2-N,N-dimethylaminoethanol under
nitrogen atmosphere in the presence of DBU (Scheme 2).
The synthesis of the metal-free phthalocyanine com-
pound (4) was accomplished by the self heating of 4-(2-
oxo-2H-chromen-7-yloxy)-phthalonitrile 2 without metal
salt at 150 °C under a N2 atmosphere in the presence of
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) for 24 h.
The characterization of the new products (3,4) involved
a combination of methods including elemental analyses,
The synthesis of the metallophthalocyanines 5–10 was
achieved by metal ion mediated cyclotetramerization
reaction of the corresponding precursors 1-2 in high boiling
dimethylaminoethanol (DMAE) using Zn(ClO4)2.6H2O,
CuCl2.2H2O and CoCl2.6H2O salts under N2 atmosphere
(Scheme 3).
1
IR, UV–vis and H NMR spectroscopies.
The IR spectrum of metal-free phthalocyanine 3 showed
a peak at 3276 cm-1 due to NH vibrations. The disap-
pearance of the CN stretching vibration on the IR spectra of
Metallophthalocyanines 8, 9, and 10 were obtained from
with
the reaction of phthalonitrile derivative
2
CH3
Co
6
H
R
M
Zn
5
Cu
Zn
8
Co
9
Cu
10
7
Compound
Scheme 3 Protocol synthesis of compound 5–10
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J Incl Phenom Macrocycl Chem
corresponding metal salts Zn(ClO4)2Á6H2O for complex 8,
CuCl2Á2H2O for complex 10, and CoCl2Á6H2O for 9 in
2-(dimethylamino) ethanol at 170 °C for 24 h. The yields
were considerably high (approximately 61 % for 8, 81 %
for 9 and 82 % for 10).
The Q bands appear at the range of 600–750 nm and
consist of one sharp and intense band and two low-intense
shoulders. The B bands appear at the range of 250–400 nm
as a broad band. In absorption spectra of metallo-ph-
thalocyanines, Q bands are originated from the electron
transitions between the HOMO and LUMO and B bands
correspond to the electron transitions between deeper
levels of occupied MO and LUMO.
The novel metallophthalocyanines are effortlessly sol-
uble in dimethyl sulfoxide (DMSO).
The UV–vis spectral data of the studied phthalocyanines
3–10 in DMSO were given in Table 1.
R
CH3
H
M
Zn
Co
Cu
Zn
Co
Cu
Compound
5
6
7
8
9
10
Antimicrobial activity
The synthesized compounds (3–10) were evaluated for
in vitro antimicrobial activity by the well diffusion method.
The antimicrobial activity of the tested compounds was
presented in Table 2 by the formation of an inhibitory
zone. In parallel, the MIC values were determinated in
Table 3. The diameter of the zone of inhibition indicates
the degree of sensitivity of the microorganisms to the
compounds the well contains. It was obtained that, five
compounds (3, 4, 5, 6 and 7) present the best antimicrobial
activity with a broad spectrum. The compounds 9 showed
antimicrobial activity against all bacteria and fungi tested
with MIC values of 0.625–5 mg/ml. The compounds 3 and
4 exhibited an important antibacterial activity against M.
luteus, S. aureus, L. monocytogenes and P. aeruginosa and
antifungal activity against C. albicans and C. tropicalis
with an important diameter ranging from 16 to 26 mm. The
Compound 5 exhibited the highest in vitro antibacterial
activity against M. luteus with MIC values of 0.312 mg/ml
while the Compound 6 showed a moderate antibacterial
activity against M. luteus, S. aureus, L. monocytogenes, S.
Typhimurium and P. aeruginosa with MIC values ranging
from 5 to 10 mg/ml.
1
Elemental analysis, H NMR, IR and UV–vis spectra
confirmed the proposed structures of the metallophthalo-
cyanines 5–10.
In the IR spectra of the phthalocyanines 5–7, the proof
of the cyclotetramerizations were the absence of the
-C:N stretching vibration observed at 2227 cm-1 of the
compounds 1. The main differences between metal-free (3,
4) and metallophthalocyanines (5–10) are inner core N–H
stretching vibrations observed at 3276, 3418 cm-1 in the
IR spectra, respectively. The rest of the spectra are similar
to that of compounds 1–2.
1
The H NMR spectra of the peripherally tetra-substi-
tuted Pcs are just about the same as those of the initial
compound, except for extension and small shifts of the
peaks. It is expected that the broadening is due to both
chemical exchange caused by aggregation-disaggregation
equilibrium in CDCl3 [30].
1H NMR Spectrum of the phthalocyanines 5, 8 provided
the characteristic chemical shifts and confirmed the pro-
posed structures. In that spectrum taken in DMSO-d6 of
comound 5, aromtic protons appeared between 8.12 and
8.45 ppm. In addition to these results, elemental analysis
These results show that the most synthesized com-
pounds have an effective antimicrobial potential against
food- borne pathogens and clinical microorganisms.
1
confirmed the proposed structure. The H NMR spectrum
of 8 exhibited characteristic signals for aromatic protons
between 8.38 and 8.50 ppm.
Compounds 6, 7, 9 and 10 have paramagnetic atoms
(Co2? and Cu2?) in the inner core. Paramagnetic com-
pounds would affect the magnetic shimming. For this
reason, paramagnetic compounds generally are not char-
Aggregation behaviours
Aggregation is usually depicted as a coplanar association
of rings progressing from monomer to dimer and higher
order complexes. It is dependent on the concentration,
nature of the solvent, nature of the substituents, complexed
metal ions and temperature [33–35]. In this study, the
aggregation behaviour of the metal-free 3 and metalloph-
thalocyanine complexes 5, are investigated in DMSO
(Schemes 4, 5).
1
acterized via NMR. The H NMR spectra of the param-
agnetic phthalocyanines 6, 7, 9 and 10 were not measured.
1
The H-NMR spectra of CuPc (7,10) and CoPc (9,6) were
excluded due to their paramagnetic properties [31].
UV–vis electronic spectra are especially practical for
identifying the structure of Pcs. Generally, for Pc com-
plexes, UV–vis spectra show typical electronic spectra with
2 strong absorption bands known as Q and Soret bands
(B) [32].
As the concentration was increased, the intensity of
absorption of the Q band corresponding to monomeric
123

J Incl Phenom Macrocycl Chem
Table 1 UV–Vis data for the
phthlocyanine compounds 3–10
Compounds
Solvent
kmax (nm) (loge/dm3 mol-1 cm-1
)
3
DMSO
DMSO
DMSO
DMSO
DMSO
DMSO
DMSO
DMSO
339 (4.729), 593 (4.535), 620 (4.633), 660 (5.005), 704 (5.066)
331 (4.531), 601 (4.201), 628 (4.408), 664 (4.758), 710 (5.010)
336 (5.044), 611 (4.642), 678 (5.197)
4
5
6
342 (4.946),628 (4.706), 668 (5.060)
7
333 (4.881), 616 (4.507), 675 (5.020)
8
340 (5.10), 634 (4.89), 679 (5.22).
9
350 (4.95), 612 (4.55), 679 (5.24).
10
342 (4.79), 626 (4.67), 672 (5.07)
species also increased and there were no new bands due to
the aggregated species for both of the complexes. Beer–
Lambert law was obeyed in the concentrations ranging
from 6 9 10-4 to 14 9 10-6M for unmetalled 3 and
metallophthalocyanines 5. It can clearly be concluded that
the phthalocyanines derivatives (3 and 5) did not show
aggregation in DMSO for the studied concentrations.
0.3 % of the theoretical values. The UV spectra were
recorded on a Perkin Elmer Lambda 11 spectrophotometer.
Synthesis of 4-(4-Methyl-2-oxo-2H-chromen-7-
yloxy)-phthalonitrile 1
The synthesis of 2 was similar to that of 1, except 7-hy-
droxy-4-methylcoumarin (1.232 g, 7 mmol) was employed
instead of 7-hydroxycoumarin. The amounts of the other
reagents were: 4-nitrophthalonitrile, 0.865 g (5 mmol) and
anhydrous potassium carbonate, 1.035 g (7.55 mmol).
Yield: (75 %). M.p = 330 °C.
Conclusion
In this research novel phthalonitriles and phthlocyanines
1
were synthesized and characterized. IR, UV–vis, H and
FT-IR (KBr) m cm-1: 3079 (Ar–CH), 2227 (CN), 1610
13C-NMR, mass spectroscopy and elemental analysis were
used to characterize the new compounds. All the com-
pounds show excellent solubility in DMSO. The aggregation
behaviour of the metal-free 3 and metallophthalocyanine
complexes 5, were investigated in DMSO and they did not
show aggregation for concentrations ranging from 6 9 10-4
to 14 9 10-6 M. Most of the new compounds 3–10 exhibited
an important antibacterial activity. The Compound 7 exhib-
ited the highest in vitro antibacterial activity against M. luteus
with MIC values of 0, 312 mg/ml while the Compound 8
showed a moderate antibacterial activity against M. luteus, S.
aureus, L. monocytogenes, and S. Typhimurium and P.
aeruginosa with MIC values ranging from 5 to 10 mg/ml.
1
(C=C), 1735 (C=O, lactone and ester), 1495 (C–O–C). H
NMR.(DMSO-d6, 300 MHz) (d: ppm) 2.42 (s, 3H, CH3),
7.43-8.11 (m, 7H, Harom ? H2,3).
13C-NMR (DMSO-d6, 75 MHz) (d: ppm) 18.5(CH3),
159.8(C2), 153.2(C4), 108.6(C3), 115.4(CN), 115.7(CN),
116.2–161.7(Carom).
Calcd for C18H10O3N2: C, 71.52; H, 3.33; N, 9.26.
Found: C, 71.5; H, 3.3; N, 9.2
MS (LCMS-MS) m/z: Calc. 302.284; Found: 302.2: m/z
[M]?.
Synthesis of 4-(2-Oxo-2H-chromen-7-yloxy)-
phthalonitrile 2
4-Nitrophthalonitrile (2.25 g, 12.34 mmol) was dissolved
in dry DMF (15 mL) under argon and 7-Hydroxycoumarin
(2.00 g, 12.34 mmol) was added. After stirring for 15 min
at room temperature, finely ground anhydrous potassium
carbonate (3.67 g, 26.6 mmol) was added in portions dur-
ing 2 h with efficient stirring. The reaction mixture was
stirred under argon atmosphere at room temperature for
24 h after which time, the ensuing mixture was poured into
100 mL iced water and the precipitate filtered off, washed
with methanol and then dried. The crude product was
chromatographered over a silica gel column using a mix-
ture of CHCl3: MeOH (100:5 (v:v)) as eluent, giving a
powder of 4-(2-Oxo-2H-chromen-7-yloxy)-phthalonitrile
1. Finally, the pure powder was dried in a vacuum. Yield:
Experimental
All reagents were obtained from Fluka and Aldrich. The
purity of the products was tested in each step by TLC
(SiO2, CHCl3/MeOH and THF/MeOH). Melting points
were determined using an Electrothermal apparatus and are
1
uncorrected. H NMR and 13C NMR spectra were carried
on a Varian Gemini 300 (300 MHz) spectrometer using
TMS as internal standard (d = 0 ppm). IR spectra were
recorded on a Perkin-Elmer 398 Spectrophotometer. MS
were recorded on a LC–MS–MS 8030 Shimadzu. Ele-
mental analyses were performed on Perkin-Elmer 2400
elemental analyzer, and the values found were within
123

J Incl Phenom Macrocycl Chem
Table 2 Antimicrobial activity spectrum of the synthesized
compound
Table 2 continued
Microrganism indicator
Compounds
7
Inhibition zone (mm)
19
Microrganism indicator
Compounds
Inhibition zone (mm)
8
16
18
14
14
13
16
16
–
Micrococcus luteus
1
2
17
16
26
26
22
14
27
18
13
20
12
11
25
23
–
9
LB 14110
10
1
3
Pseudomonas aeruginosa
4
ATCC 49189
2
5
3
6
4
7
5
8
6
12
20
18
16
13
9
9
7
10
1
8
Staphylococcus aureus
9
ATCC 6538
2
10
1
3
Candida albicans
4
2
8
5
3
22
12
–
6
14
18
13
14
16
12
10
21
20
–
4
7
5
8
6
–
9
7
10
–
10
1
8
Listeria monocytogenes
9
13
14
11
10
20
16
–
ATCC 19117
2
10
1
3
Candida tropicalis
4
R2 CIP203.
2
5
3
6
13
20
16
18
14
13
14
–
4
7
5
8
6
–
9
7
15
18
14
12
10
1
8
Agrobacterium tumefaciens
9
2
10
3
4
–
5
–
(74 %). M.p = 325 °C FT-IR (KBr) m cm-1: 3075 (Ar–
CH), 2225 (CN), 1602 (C=C), 1731 (C=O, lactone and
6
–
1
7
19
–
ester), 1496 (C–O–C). H NMR.(DMSO-d6, 300 MHz) (d:
ppm) 6.47–8.18 (m, 8H, Harom ? H3). 13C-NMR (DMSO-
d6, 75 MHz) (d: ppm) 157.3(C2), 155.4(C4), 108.2(C3),
115.6(CN), 115.7(CN), 109.8–160.2(Carom).
Calcd for C17H8O3N2: C, 70.83; H, 2.79; N, 9.71.
Found: C, 70.80; H, 2.7; N, 9.7; MS (LC–MS) m/z: Calc.
287.2; Found: 287.2
8
9
–
10
1
–
Salmonella Typhimurium
11
10
12
12
–
ATCC 14028
2
3
4
General procedure for the synthesis of metal-free 3,4
5
6
12
A mixture of 4-(4-Methyl-2-oxo-2H-chromen-7-yloxy)-
phthalonitrile
1
or 4-(2-Oxo-2H-chromen-7-yloxy)-
123

J Incl Phenom Macrocycl Chem
Table 3 Determination of the minimum inhibitory concentrations
(MICs) expressed in mg/ml
Table 3 continued
Microorganism indicator
Compounds
MIC(mg/ml)
Microorganism indicator
Compounds
MIC(mg/ml)
Candida tropicalis
3
4
2.5
5
Micrococcus luteus
3
4
5
R2 CIP203
LB 14110
5
5
–
5
0.3125
10
1.25
4
6
–
6
7
5
7
8
–
8
9
2.3
5
9
1.5
8
10
10
3
Staphylococcus aureus
5
ATCC 6538
4
5
5
–
6
5
7
2.5
5
1,4
8
1,5
1
9
4
y = 89037x + 0,030
R² = 0,996
1,2
0,5
0
10
3
–
4,00E-06
9,00E-06
1,40E-05
1,0
Concentraꢀon (M)
Listeria monocytogenes
2.5
5
6.10-6 M
8.10-6 M
10.10-6 M
12.10-6 M
14.10-6 M
ATCC 19117
4
0,8
0,6
0,4
0,2
0,0
5
–
6
5
7
0.625
–
8
9
5
10
3
3.5
5
Salmonella Typhimurium
300
400
500
600
700
800
ATCC 14028
4
10
–
Wavelength (nm)
5
Scheme 4 The aggregation behavior of Unmetalled phthalocyanine 3
in DMSO
6
10
5
7
8
–
9
5
10
3
8
Pseudomonas aeruginosa
5
1,6
ATCC 49189
4
5
1,5
1,4
1
5
–
y = 71000x + 0,412
R² = 0,998
0,5
0
4,00E-06
6
10
2.5
5
1,2
9,00E-06
1,40E-05
Concentraꢀon (M)
7
1,0
0,8
0,6
0,4
0,2
0,0
8
9
5
6.10-6 M
8.10-6 M
10.10-6 M
12.10-6 M
14.10-6 M
10
3
2.2
2.5
5
Candida albicans
4
5
–
6
–
7
–
300
400
500
600
700
800
Wavelength (nm)
8
–
9
–
Scheme 5 The aggregation behavior of metallophthalocyanine 5 in
DMSO
10
2.5
123

J Incl Phenom Macrocycl Chem
phthalonitrile 2 (0.25 g, 0.487 mmol) and catalytic amount
of 1.8-diazabicyclo[5.4.0]undec-7-ene (DBU) in 2.5 ml of
DMAE was heated and stirred at 150 Æ C in a sealed glass
tube for 24 h under N2. After cooling to room temperature
the green crude product was precipitated with ethanol,
filtered and washed first with ethanol then diethyl ether and
then dried in vacuo. Finally, pure metal-free phthalocya-
nine was obtained by column chromatography which is
placed aluminum oxide using CHCl3:CH3OH (99:1) as
solvent system.
Elution solvent system: chloroform: methanol (100:3) as
eluent.
Yield: 45 %
M.p = 330 °C
FT-IR (KBr) m, cm-1: 3066 (C–Harom); 1390 (C–C);
1280 (C–N); 1605 (C=C); 1250 (C=N); 896 (Zn–N).
Calc. for C68H32N8O12Zn: C, 67.03; H, 2.64, N, 9.19
Found: C, 67.0; H, 2.6, N 9.1; 1H NMR.(DMSO-d6,
300 MHz) (d: ppm): 2.1(s,12H,CH3),8.12–8.45 (m, 28H,
Harom ? H3,4), UV–vis (DMSO) kmax/nm: 336 (5.044),
611 (4.642), 678 (5.197).
Metal free Pc (3).
Yiled: 63 %
Co(II)Pc (6).
Elution solvent system: chloroform:methanol (100:5)as
eluent.
M.p = 335 °C
FT-IR (KBr, mmax, cm-1): 3276 (N–H); 3071 (C–
arom); 1471 (C–C); 1265 (C–N); 1602 (C=C); 1208 (C–
H
O
Yield: 71 %
lactone); 1725 (C=Olactone).
M.p = 335 °C
Calc. for C68H34N8O12: C, 70.71;H, 2.96 N,
FT-IR (KBr, mmax, cm-1): 3064 (C–Harom); 3065 (C–
Harom); 1387 (C–C); 1282 (C–N); 1608 (C=C); 1254
1
9.701Found: C, 70.7; H, 2.9, N, 9.7; H NMR.(DMSO-d6,
300 MHz) (d: ppm): 2.42(s,12H,CH3),8.32–8.42 (m, 28H,
Harom ? H3,4).
(C=N); 894 (Co–N).
Calc. for C68H32N8O12Cu: C 67.13; H, 2.65,N
9.21Found: C 67.1; H, 2.6, N,9.2; UV–vis (DMSO) kmax/
nm: 342 (4.946),628 (4.706), 668 (5.060).
Cu(II)Pc (7)
UV/Vis (DMSO, kmax nm (log e)): 339 (4.729), 593
(4.535), 620 (4.633), 660 (5.005), 704 (5.066).
Metal free Pc (4)
Elution solvent system: Chloroform: methanol (100:2)
as eluent. Yield: 113 mg (77 %).
Elution solvent system: chloroform:methanol (100:5) as
eluent.
M.p = 315 °C
Yield: 83 %
FT-IR (KBr, mmax, cm-1): 3418 (N–H); 3019 (C–
arom); 1489 (C–C); 1276 (C–N); 1612 (C=C); 1214 (C–
M.p = 325 °C
H
O
FT-IR (KBr, mmax, cm-1): 3066 (C–Harom); 1390 (C–
C); 1281 (C–N); 1610 (C=C); 1250 (C=N); 898 (Cu–N).
Calc. for C72H40N8O12Cu: C 67.949; H, 3.168, N
8.805.993 Found: C 67.9; H, 3.1, N, 8.8.
UV–vis (DMSO) kmax/nm: 333 (4.881), 616 (4.507),
675 (5.020).
lactone); 1731 (C=Olactone);
Calc. for C72H42N8O12: C, 71.401; H,3.495, N, 9.252
Found: C, 71.4; H,3.4, N, 9.1; 1H NMR.(DMSO-d6,
300 MHz) (d: ppm): 7.92-8.40 (m, 32H,Harom ? H3,4),
UV–vis (DMSO) kmax/nm: 331 (4.531), 601 (4.201),
628 (4.408), 664 (4.758), 710 (5.010);
Zn(II)Pc (8)
Elution solvent system: chloroform: methanol (100:3) as
eluent.
General procedure for the synthesis of metal-free
and metallophthalocyanines 5–10
Yield: 61 %
M.p = 330 °C
FT-IR (KBr) m, cm-1: 1400 (C–C); 1284 (C–N); 1600
(C=C); 1550 (C=N); 896 (Zn–N).
Calc. for C72H40N8O12Zn: C, 67.850; H, 3.163, N,
Compounds (1) or (2) (0.24 mmol), dry N,N-dimethy-
laminoethanol
(DMAE)
(4 ml),
1,8-diazabicyclo
1
[4.5.0]undec-7-ene (DBU) (3 drops) and (0.06 mmol) of
corresponding anhydrous metal salts (Zn(ClO4)2Á6H2O,
CuCl2Á6H2O, CoCl2Á6H2O and NiCl2Á6H2O) were added in
schlenk tube. The mixture was heated at reflux temperature
of 170 °C for 24 h under N2 atmosphere. Then the mixture
was left for cooling at room temperature then treated with
ethylacetate to precipitate the product which was then fil-
tered off and suddenly washed with water. The green solid
product was washed with hot ethanol and dried in vacuo.
The raw product was purified by chromatography of silica
gel column.
8.792 Found: C, 67.8; H, 3.1, N 8.7; H NMR.(DMSO-d6,
300 MHz) (d: ppm): 8.38-8.50 (m, 24H, Ar–H).
UV–vis (DMSO) kmax/nm: 340 (5.10), 634 (4.89), 679
(5.22).
Co(II)Pc (9). Elution solvent system: chloro-
form:methanol (100:2) as eluent. Yield: 114 mg (81 %).
mp = 338 °C.
FT-IR (KBr) m, cm-1: 3065 (C–Harom); 1406 (C–C);
1282 (C–N); 1598 (C=C); 1558 (C=N); 897 (Co–N).
Calc. for C68H32N8O12Co: C 67.38, H, 2.66, N 9.24,
Found: C 67.3, H, 2.6, N 9.2. UV–vis (DMSO) kmax/nm:
350 (4.95), 612 (4.55), 679 (5.24).
Zn(II)Pc (5).
123

J Incl Phenom Macrocycl Chem
Cu(II)Pc (10)
Elution solvent system: chloroform:methanol (100:5) as
eluent.
be a confluent lawn of growth. The antibacterial activity
was assayed by measuring in millimeters the diameter of
the inhibition zone formed around the well.
Yield: (82 %)
mp = 325 °C
MIC determination
FT-IR (KBr) m, cm-1: 1404 (C–C); 1284 (C–N); 1604
(C=C); 1554 (C=N); 896 (Cu–N).
Calc. for: C72H40N8O12Co: C 68.196; H, 3.179, N 8.837;
Found: C 68.2; H, 3.1, N, 8.8.
UV–vis (DMSO) kmax/nm: 342 (4.79), 626 (4.67), 672
(5.07).
The antimicrobial activities of the synthesized compounds
were determined by the minimum inhibitory concentration
(MICs) in accordance with NCCLS guideline M7-A6 and
M38-P [37]. The test was performed in sterile 96-well
microplates with a final volume in each microplate well of
100 ll. The synthesized compounds (20 mg/mL) were
properly prepared in solution of dimethylsulfoxide (20 %).
The inhibitory activity of each synthesized compounds was
transferred to each well in order to obtain a twofold serial
dilution of the original sample and to produce the con-
centration range of 0.0781–10 mg/mL.
Antimicrobial activities
Microorganisms and growth conditions
Bacterial strains: Gram-positive bacteria (Micrococcus
luteus LB 14110, Staphylococcus aureus ATCC 6538,
Listeria monocytogenes ATCC 19117 and Agrobacterium
tumefaciens), Gram-negative bacteria (Salmonella Typhi-
murium ATCC 14028 and Pseudomonas aeruginosa
ATCC 49189) were used as indicator microorganisms for
the antibacterial activity assays. Antifungal activity was
determined against the Candida albicans and Candida
tropicalis R2 CIP203.
To each test well 10 lL of cell suspension were added
to final inoculum concentrations of 106 CFU/ml for each
microorganisms. Positive growth control wells consisted
of microorganisms only in their adequate medium. Cells
suspension at the same concentration supplemented with
ampicillin (50 lg/mL) was used as negative control. The
plates were then covered with the sterile plate covers and
incubated at the appropriate temperature of each
microorganism. The MIC was defined as the lowest
concentration of the synthesized compounds at which the
microorganism does not demonstrate visible growth after
incubation. As an indicator of microorganism growth,
25 ll of Thiazolyl Blue Tetrazolium Bromide (MTT),
indicator solution (0.5 mg/ml) dissolved in sterile water
were added to the wells and incubated at room temper-
ature for 30 min.
The bacterial strains using as indicator microorganisms
were grown overnight in LB media under aerobic condi-
tions and constant agitation (200 rpm) at 30 °C for M.
luteus LB14110, L. monocytogenes ATCC 19117, and A.
tumefaciens, and at 37 °C for S. aureus ATCC 6538, S.
Typhimurium ATCC 14028 and P. aeruginosa ATCC
49189, and then diluted 1:100 in LB media and incubated
for 5 h under constant agitation (200 rpm) at the appro-
priate temperature. C. tropicalis R2 CIP203 was grown in
YP10 medium (10 g/l yeast extract, 10 g/l peptone, 100 g/l
glucose, 15 ml of 2 g/l adenine solution) and C. albicans
was grown in YEPD medium (10 g/l yeast extract, 20 g/l
peptone, 20 g/l dextrose) at 30 °C for 48 h in an orbital
incubator with shaking at 200 rpm.
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