Crystal structures and antifungal activities of fluorine-containing thioureido complexes with nickel(II)

Article abstract of DOI:10.3390/molecules181215737

Ni(II) complexes with N-2-fluorobenzoylpiperidine-1-carbothioimidate (L2-), N-4-fluorobenzoylpiperidine-1-carbothioimidate (L3-), N-2- fluorobenzoylmorpholine- 1-carbothioimidate (L5-) and N-4- fluorobenzoylmorpholine-1-carbothioimidate (L6-) have been synthesized and characterized by elemental analysis, FTIR and 1H-NMR. The crystal structures of three ligands (HL2, HL3 and HL6) and the corresponding Ni(II) complexes ([Ni(L2)2], [Ni(L3)2] and [Ni(L6)2]) have been determined by X-ray diffraction. The antifungal activities of the Ni(II) complexes together and the corresponding ligands against the fungi Botrytis cinerea, Trichoderma spp., Myrothecium and Verticillium spp. have been investigated. The experimental results showed that the ligands and their complexes have antifungal abilities. When the fluorine was substituted on the para-benzoyl moiety, the antifungal activity of the ligands was obviously increased. Moreover, the ligands were stronger than their complexes in inhibiting fungal activities. The antifungal ability of HL6 is especially strong, and similar to that of the commercial fungicide fluconazole.

Full text of DOI:10.3390/molecules181215737

Molecules 2013, 18, 15737-15749; doi:10.3390/molecules181215737
OPEN ACCESS
molecules
ISSN 1420-3049
www.mdpi.com/journal/molecules
Article
Crystal Structures and Antifungal Activities of
Fluorine-Containing Thioureido Complexes with Nickel(II)
Chun Li 1,2, Wen Yang 2, Huanhuan Liu 2, Mengying Li 3, Weiqun Zhou 2,* and Juan Xie 2
1
College of Pharmacy, Nantong Tichen Health School, Nantong 226007, China
2
College of Chemistry, Chemical Engineering and Materials Science, Soochow University,
199 Ren’ai Road, Suzhou 215123, China
3
School of Biology & Basic Medical Sciences, Soochow University, 199 Ren’ai Road,
Suzhou 215123, China
* Author to whom correspondence should be addressed; E-Mail: wqzhou@suda.edu.cn;
Tel.: +86-512-6588-4827.
Received: 13 November 2013; in revised form: 3 December 2013 / Accepted: 5 December 2013 /
Published: 17 December 2013
Abstract: Ni(II) complexes with N-2-fluorobenzoylpiperidine-1-carbothioimidate (L2−),
N-4-fluorobenzoylpiperidine-1-carbothioimidate (L3−), N-2-fluorobenzoylmorpholine-
1-carbothioimidate (L5−) and N-4-fluorobenzoylmorpholine-1-carbothioimidate (L6−)
1
have been synthesized and characterized by elemental analysis, FTIR and H-NMR.
The crystal structures of three ligands (HL2, HL3 and HL6) and the corresponding Ni(II)
complexes ([Ni(L2)2], [Ni(L3)2] and [Ni(L6)2]) have been determined by X-ray diffraction.
The antifungal activities of the Ni(II) complexes together and the corresponding ligands
against the fungi Botrytis cinerea, Trichoderma spp., Myrothecium and Verticillium spp.
have been investigated. The experimental results showed that the ligands and their
complexes have antifungal abilities. When the fluorine was substituted on the para-benzoyl
moiety, the antifungal activity of the ligands was obviously increased. Moreover, the ligands
were stronger than their complexes in inhibiting fungal activities. The antifungal ability of
HL6 is especially strong, and similar to that of the commercial fungicide fluconazole.
Keywords: fluorobenzoyl thioureas; Ni(II) complexes; crystal structures; antifungal activity

Molecules 2013, 18
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1. Introduction
As we know, pyridine, morpholine and thiourea exhibit broad biological activities, including
antibacterial and antifungal effects [1–11]. 1,3-Dialkyl- or diarylthioureas present significant antifungal
activity against the plant pathogens Pyricularia oryzae and Drechslera oryzae [12]. Acyl thioureas are
well known for their superior pesticidal, fungicidal, antiviral and plant growth regulating activity [13].
Moreover, thioureas containing both carbonyl and thiocarbonyl groups are versatile ambidentate donor
ligands for transition metal ions [14].
The introduction of fluorine at a strategic position of a molecule is a powerful and versatile tool for
the development of organic molecules which have potential biological activities by changing the steric
and electronic parameters [15–21]. The inclusion of fluorine into organic molecules can increase the
lipophilicity and thus enhance the rate of cell penetration and transport of a drug to an active site [22].
Fluorinated thioureas constitute a novel class of potent influenza virus neuraminidase inhibitors.
The biological activities of metal complexes are different from those of either their ligands or the free
metal ions. In many cases, upon coordination to metal ions, the bioactivity of these complexes increases,
suggesting that complexation can be an interesting dose reduction strategy [23–28]. However, some
Co(III) and Ni(II) complexes had the weaker antibacterial activities than the corresponding ligands [11,29].
The proximate experiments of our group on fluorobenzoylthioureas and their Co(III) complexes [29] have
revealed the introducing a fluorine atom on para-benzoylthiourea increased the antibacterial activities,
while when the fluorine atom was substituted on an ortho-benzoyl group, the antibacterial activity
was weakened.
In this paper, we synthesized four complexes, [Ni(L2)2], [Ni(L3)2], [Ni(L5)2] and [Ni(L6)2].
The crystal structures of the three ligands (HL2, HL3, HL6) and the corresponding complexes were
determined by X-ray diffraction. In consideration of the knwon good biological effects of nickel ion and
its complexes [30,31], we were interested in exploring the effect of fluorine substitution and the
coordination with Ni (II) on the antifungal activities.
2. Results and Discussion
2.1. Structural Characteristic
The molecule structures of the three ligands and the corresponding complexes with Ni(II) are shown
in Figures 1–3, and their X-ray diffraction parameters are summarized in Table 1. Three ligands show
trans-conformation between the C=S and C=O. No significant differences appear in the bond lengths
and bond angles of the three ligands (Table 3). The amidic C(1)-N(1), thiomide N(2)-C(8) and
N(1)-C(8) bonds are shorter than the C–N single bond length of 1.472(5) Ǻ. The partial double bond
character of the structure is presumed as a result of the p-π conjugation. The length of the bond
C(8)-S(1) lies between the values of C–S single and double bonds. In the crystals of HL3, two
molecules have some small conformational distinctions.

Molecules 2013, 18
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Figure 1. Molecular structures of HL2 and [Ni(L2)2], with thermal ellipsoids drawn at
40% probability.
HL2
[Ni(L2)2]
Figure 2. Molecular structures of HL3 and [Ni(L3)2], with thermal ellipsoids drawn at 40% probability.
HL3
[Ni(L3)2]

Molecules 2013, 18
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Figure 3. Molecular structures of HL6 and [Ni(L6)2] with thermal ellipsoids drawn at 40% probability.
HL6
[Ni(L6)2]
Table 1. Summary of X-ray diffraction data.
Parameter
HL2
[Ni(L2)2]
HL3
[Ni(L3)2]
HL6
[Ni(L6)2]
Empirical formula C13H15FN2OS C26H28F2N4NiO2S2 C13H15FN2OS C26H28F2N4NiO2S2 C12H13FN2O2S C24H24F2N4NiO4S2
Formula weight
Crystal system
Space group
a (Å)
266.33
Monoclinic
P 21/c
589.35
Orthorhombic
P b c a
11.6111(17)
9.8778(17)
45.925(7)
90
266.33
Triclinic
P-1
589.35
Triclinic
P-1
268.3
Monoclinic
C 2/c
593.3
Triclinic
P-1
11.843(3)
13.282(3)
8.432(2)
90
8.3759(15)
11.191(2)
13.960(3)
89.981(10)
89.969(10)
79.699(9)
4
9.5530(16)
11.890(2)
12.548(2)
104.350(4)
100.377(3)
97.110(4)
2
21.214(6)
9.7391(16)
14.637(7)
90
9.3383(6)
11.5273(9)
12.7230(8)
105.206(6)
101.420(5)
96.112(6)
2
b (Å)
c (Å)
α (°)
β (°)
108.474(5)
90
90
125.91(3)
90
γ (°)
90
Z
4
8
8
Dcalc (g/cm3)
Radiation (MoKα)
(Å)
1.406
1.486
1.374
1.464
1.455
1.543
0.71075
0.7107
0.7107
0.7107
0.71075
0.71073
μ(Mo Kα) (mm−1)
θ Range (°)
Reflections
collected
0.259
0.941
0.253
0.927
0.272
0.977
3.03 to 27.50
3.02 to 25.35
3.17 to 25.34
3.01 to 25.35
3.34 to 27.50
3.03 to 26.37
7538
2850
34,645
4799
12,652
4678
12,987
4872
10,790
2773
13,024
5220
Independent
reflections
R(int)
0.022
2850
0.0687
4799
0.0719
4678
0.0292
4872
0.0514
2773
0.0208
5220
Data
restraints
0
0
2
0
0
17
Parameters
GOF on F2
R1 [I > 2sigma(I)]
wR2 [I > 2sigma(I)]
R1 [all data]
wR2 [all data]
Largest difference
peak (e.A−3)
165
344
334
335
163
334
1.011
0.0436
0.1141
0.0551
0.1222
0.362 and
−0.377
1.009
0.0904
0.1785
0.11
1.098
1.079
0.999
1.023
0.0838
0.1771
0.1575
0.2080
0.265 and
−0.316
0.0536
0.1242
0.0723
0.1353
0.627 and
−0.297
0.0422
0.1084
0.0636
0.1190
0.331 and
−0.449
0.0406
0.1002
0.0564
0.1105
0.552 and
−0.315
0.1892
0.728 and
−0.413

Molecules 2013, 18
15741
The crystal structure of [Ni(L2)2] shows static disorder for the fluorine atoms on one phenyl ring so
the positions of atoms cannot be determined with a sufficient accuracy. All of complexes exhibit
cis-tetracoordinate structures around nickel(II) in the crystalline molecules, which are similar to those of
[Ni(L4)2] [32], cis-bis(1,1-diethyl-3-benzoylthiourea)-nickel(II) [33] and cis-bis(1,1-diethyl-3-(2-
chlorobenzoylthiourea-O,S)-nickel(II) [34]. The slightly distorted square geometry is also found in the
structure of the three complexes. The O–Ni–O, S–Ni–S and O–Ni–S angles deviate slightly from 90.0°.
The conformation isomerizes, which must have happened during the coordination process. The
C1–N1–C8–S1 and C14–N3–C21–S2 torsion angles (Table 2) corresponding to the cis-conformation
are completely different from the trans-conformations of the ligands.
Table 2. Some structure parameters of the determined crystals
Distances (Å)
Ni(1)-O(1)
Ni(1)-O(2)
Ni(1)-S(1)
Ni(1) -S(2)
S(1)-C(8)
HL2
[Ni(L2)2]
1.859 (4)
1.846 (4)
2.130 (2)
2.147 (2)
HL3
[Ni(L3)2]
1.864 (2)
1.866 (2)
2.140 (11)
2.139 (11)
1.742 (4)
1.731 (4)
HL6
[Ni(L6)2]
1.861 (18)
1.864 (18)
2.145 (8)
2.142 (8)
1.737 (2)
1.673 (15) 1.728 (7)
1.723 (6)
1.668 (5)
1.666 (5)
1.679 (2)
1.219 (2)
1.392 (2)
1.397 (2)
S(2)-C(21)
S(2)-C(20)
O(1)-C(1)
O(2)-C(14)
O(2)-C(13)
N(1)-C(1)
N(3)-C(14)
N(3)-C(13)
N(1)-C(8)
N(3)-C(21)
1.731 (3)
1.268 (3)
1.215 (19) 1.271 (7)
1.269 (7)
1.233 (5)
1.233 (6)
1.275 (4)
1.268 (4)
1.272 (3)
1.321 (3)
1.378 (18) 1.319 (7)
1.309 (7)
1.358 (6)
1.358 (6)
1.312 (4)
1.319 (5)
1.307 (3)
1.342 (3)
1.413 (19) 1.337 (8)
1.341 (8)
1.434 (6)
1.430 (6)
1.340 (5)
1.334 (5)
The conformational disposition in the complexes must be derived from the electron delocalization
along the S1–C8–N1–C1–O1 fragment. The N1–C1, N3–C14, N1–C8 and N3–C21 distances are shorter
than the N1–C1 and N1–C8 distances in the ligands. The distances of N2–C8 and N4–C21 in the
complexes are longer than the distance of N2–C8 in the ligands. As Table 2 shows, all N–C bond
distances are between a C–N single bond (1.468 Å) and double bond (1.25 Å). The O–C and S–C bonds
in the complexes are longer than those in the ligands.
2.2. Antifungal Activities
The average values of the minimum inhibitory concentration (MIC) of the ligands and their
complexes against the fungal strains are shown in Table 3. Taking the commercial fluconazole, the
ligands N-(piperidine-1-thioyl)benzamide (HL1), N-(morpholine-4-thioyl)benzamide (HL4) and their
complexes, [Ni(L1)2] and [Ni(L4)2] as reference, the ligands and the complexes containing fluorine
atoms had good performance inhibiting the growth of the fungi. Especially, the antifungal abilities of
HL6 are equal to that of commercial fluconazole; the MIC values against the fungi Botrytis cinerea,
Trichoderma spp., Myrothecium and Verticillium spp. are 8.0, 8.5, 7.0 and 8.5 μmol/L, while those of

Molecules 2013, 18
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fluconazole are 7.0, 8.0, 7.5 and 7.5 μmol/L, respectively. Moreover, four ligands also have the stronger
antifungal abilities than the corresponding complexes. For example, the MIC values for [Ni(L6)2] are
31.0 (Botrytis cinerea), 25.0 (Trichoderma spp.), 27.5 (Myrothecium) and 27.5 μmol/L (Verticillium
spp.), respectively, much higher than that of HL6 (mentionedabove). The results are similar to the
antibacterial activities [24]. The inhibitory abilities of the piperidinyl ligands are inferior to those of
morpholine ligands. The MIC values for HL3 were 20.5 (Botrytis cinerea), 18.0 (Trichoderma spp.),
22.5 (Myrothecium) and 25.0 μmol/L (Verticillium spp.), respectively, which differ from the
antibacterial activities [24]. The position effect is more apparent. The introduction of a F atom at the
para-position of the benzoyl increased the antifungal activities notably. Contrarily, when the F atom
substitutes the benzoyl ortho-position, the antifungal abilities decrease.
Table 3. The MIC of the ligands and their complexes against the studied fungi (SD: standard deviation).
Samples
Fluconazoleole
HL1
Botrytis cinerea Trichoderma spp. Myrothecium Verticillium. spp.
7.0 ± 0.1
31.0 ± 0.0
33.0 ± 0.0
27.5 ± 0.0
31.0 ± 0.1
20.5 ± 0.1
22.5 ± 0.0
2.5 ± 0.1
8.0 ± 0.1
22.5 ± 0.1
31.5 ± 0.1
29.0 ± 0.0
33.0 ± 0.1
18.0 ± 0.1
31.0 ± 0.0
2 ± 0.0
7.5 ± 0.1
32.5 ± 0.1
36.0 ± 0.0
27.5 ± 0.1
31.0 ± 0.1
22.5 ± 0.1
29.0 ± 0.1
2.3 ± 0.0
7.5 ± 0.1
34.0 ± 0.0
38.0 ± 0.1
28.0 ± 0.0
31.5 ± 0.1
25.0 ± 0.0
31.0 ± 0.0
0.8 ± 0.1
[Ni(L1)2]
HL2
[Ni(L2)2]
HL3
[Ni(L3)2]
HL4
[Ni(L4)2]
HL5
29.0 ± 0.1
31.0 ± 0.1
32.5 ± 0.1
8.0 ± 0.0
32.5 ± 0.1
27.5 ± 0.0
31.0 ± 0.1
8.5 ± 0.0
25.0 ± 0.0
35.5 ± 0.1
27.5 ± 0.0
31.0 ± 0.1
7.0 ± 0.1
35.5 ± 0.1
31.0 ± 0.1
31.5 ± 0.0
8.5 ± 0.0
[Ni(L5)2]
HL6
[Ni(L6)2]
31.0 ± 0.0
27.5 ± 0.1
27.5 ± 0.0
2.3. Structure-Activity Relationships
On the one hand, organic molecules containing F have the good lipophilicity and thus enhanced rates
of cell penetration and transport to an active site [23]. Furthermore, the fluorination adjacent to a
π-system or heteroatom-containing functional group can strongly polarize the parent molecule [35].
Taking morpholine and piperidine as the donors and F atoms as the acceptors, the ligands containing F
would form a D-A molecular system. The D-A model molecules will be more active in the interaction
with biomolecules than the common, so the molecules should have strong antimicrobial activity.
The change of the antimicrobial activity is mostly attributed to the change of the structures in the
ligands and the complexes. The change in the structures must result in the change of the charge
population on the atoms of the complexes and accordingly induce the differences of antimicrobial
activities. In our recent work, it was found that the lower the LUMO energy was, the more polar the C–N
of the acyl was, and the higher the antibacterial activity was [29]. In the crystal structures of HL2, HL3
and HL6, the average C–N bond of the acyl measures 1.374, 1.368 and 1.392 Å, respectively. HL6 has
the longest C–N bond, so the C–N polarity is the largest and the antimicrobial activity must be the
strongest. Upon coordination, the C–O and C–S bonds weaken and the amide C–N bonds gradually
strengthen. The average C–N bond distances of 1.319(7), 1.312(4) and 1.321(3) Å in [Ni(L2)2],

Molecules 2013, 18
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[Ni(L3)2] and [Ni(L6)2] are shorter than those of the C–N bond of the amide, 1.378(18), 1.358(6) and
1.392(2) Å in the corresponding free ligands, HL2, HL3 and HL6. The shortened bond length of the
amide C–N bond is a disadvantage for the polarity increase, so the antifungal activity of the complexes
will decrease.
For HL2, the intramolecular hydrogen bonding N–H···F will decrease the polarity of the molecules,
and is also disadvantage for the polarity increase of the C–N bond of the amide. For HL3, no
intramolecular hydrogen bonding interactions exist, but, a dimer is formed in the crystals of HL3 due to
the strong intermolecular N–H···O hydrogen bonding interactions, with a H···O distance of 2.118, N···O
distance of 2.965Ǻ, and ∠N–H···O of 173.8°. This hydrogen bonding results in a longer C–O bond
(1.233 Ǻ), which disturbs the polarity of amide C–N bond. In the crystal structure of HL6, because the
O atom of morpholine replaces the carbon atom of piperidine, two weak intermolecular N–H···O(1)
interactions with H···O distances of 2.423 and 2.922, N···O distances of 3.408 and 3.104 Ǻ, and
∠N–H···O, 125.3° and 135.4° are observed instead of the strong N–H···O intermolecular hydrogen
bonding interactions in HL3 (Figure 4). The weak interactions do not affect the polarity of the amide
C–N bond, so HL6 has the strongest antifungal activity.
Figure 4. The packing diagrams of the corresponding hydrogen bonds of ligands.
HL2
HL3
HL6

Molecules 2013, 18
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3. Experimental
3.1. Materials
Melting points were measured with Kofler melting point apparatus (Tallmadge, OH, USA) and
were uncorrected. IR spectra were obtained in KBr discs using a Nicolet 170SX FT-IR spectrometer
1
(Thermo Fisher Scientific, Shanghai, China). H-NMR (400.13 MHz) spectra were recorded in
chloroform-d (CDCl3) solution at room temperature on an INOVA 400 instrument (VARIAN, Palo
Alto, CA, USA) with TMS as internal reference. Elemental analyses were performed on a Yanaco
CHNSO Corder MT-3 analyzer (PerkinElmer, Waltham, MA, USA). X-ray diffraction determination
was carried out with a Rigaku Mercury CCD diffractometer (Rigaku, Japan).
3.2. Synthesis
The synthetic route to the target ligands given in the literature [28] was used, as outlined in Scheme 1.
Fluorobenzoyl chlorides were obtained by the acylation reaction of the corresponding fluorobenzoic
acids with thionyl chloride.
Scheme 1. Synthetic route of the ligands and the complexes.
O
SOCl2
C
Cl
COOH
R1
R
R
O
C
KSCN
NCS
O
C
S
HN
R2
NH C
R1
HN
O
N
N
N
O
O
O
HL4: R1=-H, R2=
HL5: R1=o-F, R2=
HL1: R1=-H, R2=
HL2: R1=o-F, R2=
N
N
N
HL6: R1=p-F, R2=
HL3: R1=p-F, R2=
(C2 H7 )3 N
2[(C2 H7 )3 NH]+
[Ni(Ln)2 ]
Ni2+ + 2HLn
+
(n=2,3,5,6)
Yie ld:
[Ni(L2)2]
[Ni(L3)2]
[Ni(L5)2]
80%
82%
85%
81%
[Ni(L6)2]
Then, a freshly prepared acetone solution of benzoyl isothiocyanate synthesized by the
condensation of fluorobenzoyl chloride with KSCN was added dropwise to a stoichiometric amount of
morpholine or piperidine solution with stirring. The mixture was heated and refluxed for 1–2 h. After
that, the mixture was poured on crushed ice with vigorous stirring. The isolated solids was filtered,

Molecules 2013, 18
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washed with water and dried to yield the target ligands. The synthees of the complexes are also
depicted in Scheme 1. The complexes were obtained by the reaction of anhydrous NiSO4 with the
corresponding ligands (molar ratio of Ni2+:L = 1:2) in ethanol in the presence of triethylamine. All of
ligands and complexes are air-stable enough to be used for the antimicrobial property studies.
[Ni(L2)2]: Dark green solid, m.p.:186.5–188.4 °C. Anal. Calc. for C26H28F2N4NiO2S2 (% C, H, N and
S): 52.97, 4.75, 9.51 and 10.87. Found (% C, H, N and S): 52.92, 4.81, 9.46 and 10.92. FTIR (cm−1):
3016 (w,υPh-H), 2938 (w, υCH2), 2857 (w, υCH2), 1585 (w, υPh-H), 1526 (m, υPh-H), 1489 (m, υPh-H), 1414
1
(m, δCH2), 1358 (w, υC-F), 1244 (w, υC-O), 1132 (w, υC-N), 1020 (w, υC-S), 758 (w, δPh-H). H-NMR
(ppm): 7.96–7.93 (t, 2H, J = 6.8, 6.4 Hz), 7.38 (s, 2H), 7.14–7.10 (t, 2H, J = 7.2, 7.2 Hz), 7.05–7.00
(t, 2H, J = 9.6, 9.2 Hz), 4.06 (s, 2H), 3.96 (s, 2H), 3.96 (s, 4H), 1.68–1.61 (d, 12H, J = 28.8 Hz).
[Ni(L3)2]: Dark green solid, m.p.: 238.9–240.6 °C. Anal. Calc. for C26H28F2N4NiO2S2 (% C, H, N and
S): 52.97, 4.75, 9.51 and 10.87. Found (% C, H, N and S): 52.95, 4.80, 9.54 and 10.80. FTIR (cm−1):
3030 (w, υPh-H), 2941 (w, υCH2), 2860 (w, υCH2), 1601 (w, υPh-H), 1518 (m, υPh-H), 1483 (m, υPh-H), 1424
(m, δCH2), 1364 (w, υC-F), 1244 (w, υC-O), 1150 (w, υC-N), 1018 (w, υC-S), 856 (w,δPh-H). 1H-NMR (ppm):
8.10 (s, 4H), 7.05 (s, 4H), 4.08–3.98 (d, 8H, J = 24 Hz), 1.69 (s, 12H).
[Ni(L5)2]: Brown solid, m.p.: 229.6–230.0 °C. Anal. Calc. for C24H24F2N4NiO4S2 (% C, H, N and S):
48.57, 4.05, 9.44 and 10.79. Found (% C, H, N and S): 47.90, 4.10, 9.42 and 10.68. FTIR (cm−1): 3076
(w, υPh-H), 2923 (w, υCH2), 2856 (w, υCH2), 1589 (w, υPh-H), 1521 (m, υPh-H), 1473 (m, υPh-H), 1419 (w,
δCH2), 1356 (w, υC-F), 1252 (w,υC-O), 1110 (w,υC-N), 1020 (w,υC-S), 755 (w,δPh-H). 1H-NMR (ppm): 7.97
(d, 2H, J = 0.8 Hz), 7.44 (d, 1H, J = 2.0 Hz), 7.18–7.15 (t, 2H, J = 6.0, 7.2 Hz), 7.09–7.05 (t, 2H,
J = 8.8, 8.4 Hz), 4.18–4.05 (d, 6H, J = 24 Hz), 3.78–3.74 (d, 6H, J = 16.4 Hz), 2.99–2.91 (d, 2H,
J = 29.6 Hz), 1.61(s, 2H).
[Ni(L6)2]: Pink solid, m.p.: 302.9–303.1 °C. Anal. Calc. for C24H24F2N4NiO4S2 (% C, H, N and S):
48.57, 4.05, 9.44 and 10.79. Found (% C, H, N and S): 48.63, 3.99, 9.42 and 10.82. FTIR (cm−1): 3046
(w, υPh-H), 2968 (w, υCH2), 2856 (w, υCH2), 1601 (w, υPh-H), 1515 (m, υPh-H), 1474 (m, υPh-H), 1419 (m,
1
δCH2), 1350 (w, υC-F), 1260 (w, υC-O), 1150 (w, υC-N), 1028 (w, υC-S), 852 (w, δPh-H). H NMR (CDCl3)
(ppm): 8.21 (s, 4H), 7.08–7.41 (t, 4H, J = 8.4, 8.4 Hz), 4.17–4.16 (d, 4H, J = 4.4 Hz), 4.05–4.04
(d, 4H, J = 4.4 Hz), 3.77–3.76 (d, 4H, J = 4.4 Hz), 2.18 (s, 2H), 1.62 (s, 2H).
3.3. X-ray Structure Determination
Crystals suitable for the X-ray diffraction determination were prepared by solvent evaporation
method in dimethylformamide (DMF). All measurements were operated on a Rigaku Mercury CCD
X-ray diffractometer [36] by using graphite monochromated MoKα radiation (λ = 0.71070 Å). Single
crystals were mounted with grease at the top of a glass fiber. Cell parameters were refined on all observed
reflections by using the program CrystalClear (Rigaku and MSC, Ver. 1.3, 2001) [37]. The collected
data were reduced by the program CrystalClear and an absorption correction (multiscan) was applied.
The reflection data of the crystals were corrected by Lorentz and polarization effects. The crystal
structures were solved by direct methods and refined by full-matrix least-squares methods (F2) using the

Molecules 2013, 18
15746
SHELXTL software package [38,39]. Non-hydrogen atoms were refined anisotropically and hydrogen
atoms were placed at calculated positions. The structure of [Ni(L2)2], involved one orientation disorder,
and the molecule was refined in two positions, the main position of the atom F1A was 56.8 % and the
secondary position of the atom F1B was 43.2%. The summary of the key crystallographic information
and the selected bond lengths and bond angles for both crystals was listed in Tables 2–3 respectively.
The other information of the crystals had been deposited in the Cambridge Crystallographic Data Centre
as CCDC 836,547 (HL2), CCDC 845,010 (HL3), CCDC 877,566 (HL6), 875,172 ([Ni(L2)2]), 878,379
([Ni(L3)2]) and 882,281 ([Ni (L6)2]). These data can be obtained free of charge from The Cambridge
Crystallographic Data Centre via www.ccdc.cam.ac.uk/data_request/cif.
3.4. Determination of the Minimum Inhibitory Concentration (MIC)
The MIC of the test samples was determined by a two-fold serial dilution method. We pre-cultured
the studied fungi for 24 h at 28 °C in LB (Luria-Bertani) liquid medium (tryptone 10 g, yeast extract
5 g, NaCl 10 g and 500 mL of distilled water). Then, taking the same concentration of the studied fungi
cultured for 24 h in LB liquid medium. Subsequently, the different concentrations of the test samples
were added into the LB liquid medium containing the studied fungi and cultured for 24 h at 28 °C
(dimethyl sulfoxide (DMSO) as the control and fluconazole as the reference). The culture solution
(50 μL) was inoculated on the LB solid medium to observe the growth of the colony and the MIC was the
highest dilution concentration of no growth of the fungi. Each concentration repeated three times for each
fungus. Differences between the experimental group and the control group were analyzed by Least
Significant Difference (LSD) t-test. p < 0.05 was considered to indicate a statistically significant
difference. In this work, all results of experiments were measured up to p < 0.05.
4. Conclusions
Four fluorine-containing thiourea complexes ([Ni(L2)2], [Ni(L3)2], [Ni(L5)2], [Ni(L6)2]) have been
successfully synthesized. The crystal structures of HL2, HL3, HL6 and the nickel complexes have been
determined by X-ray diffraction. The [Ni(L2)2], [Ni(L3)2] and [Ni(L6)2] complexes have similar
cis-tetracoordinate structures.
The antifungal activity assays show the introducing fluorine in the para-position of the benzoyl
moiety in ligands is a very powerful and versatile tool to increase antifungal activities by changing the
electronic parameters and forming a D-A system. The presence of intra- and intermolecular hydrogen
bonding interactions and the coordination will be disadvantage for increasing the polarity of the amide
C-N bond and this decreases the antifungal activities, so the antifungal activity of HL6 is the strongest
among the studied samples, and is the same as that of the commercial fungicide fluconazole.
Acknowledgments
This work was supported by the National Natural Science Foundation of China (No. 51079094) and
the Natural Science Foundation of Jiangsu Province (No. BK2010215).

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Conflicts of Interest
The authors declare no conflict of interest.
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Sample Availability: Samples of the HL2, HL3, HL6, [Ni(L2)2], [Ni(L3)2] and [Ni(L6)2] are available from
the authors.
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distributed under the terms and conditions of the Creative Commons Attribution license
(http://creativecommons.org/licenses/by/3.0/).