Synthesis, antibacterial, anticancer and molecular docking studies of macrocyclic metal complexes of dihydrazide and diketone

Accepted Manuscript Original article Synthesis, antibacterial, anticancer and molecular docking studies of macrocyclic metal complexes of dihydrazide and diketone Sabir Ali, Vandna Singh, Preeti Jain, Vishwas Tripathi PII: DOI: Reference:

S1319-6103(18)30048-6 https://doi.org/10.1016/j.jscs.2018.04.005 JSCS 959

To appear in:

Journal of Saudi Chemical Society

Received Date: Accepted Date:

28 December 2017 10 April 2018

Please cite this article as: S. Ali, V. Singh, P. Jain, V. Tripathi, Synthesis, antibacterial, anticancer and molecular docking studies of macrocyclic metal complexes of dihydrazide and diketone, Journal of Saudi Chemical Society (2018), doi: https://doi.org/10.1016/j.jscs.2018.04.005

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Synthesis, antibacterial, anticancer and molecular docking studies of macrocyclic metal complexes of dihydrazide and diketone

Sabir Ali a*, Vandna Singh a, Preeti Jain b and Vishwas Tripathi c

a - Department of Applied Chemistry, School of Vocational Studies and Applied Sciences, Gautam Buddha University, Greater Noida, Uttar Pradesh - 201310, India. b - School of Basic Sciences and Research, Sharda University, Greater Noida, Uttar Pradesh 201310, India. c – School of Biotechnology, Gautam Buddha University, Greater Noida, Uttar Pradesh 201310, India.

*Corresponding author: Tel: 0120-2344393, E-mail: [email protected]

Abstract The complexes of Co(II), Ni(II), Cu(II) and Zn(II) metal ions has been synthesized through template method

by the condensation of succinic acid dihydrazide with 5-chloroisatin in

alcoholic medium. Complexes were characterized by C H N analysis, molar conductance, thermal analysis, magnetic susceptibility, mass spectrometry, FTIR, EPR, 1H NMR, UV-Visible spectroscopy. These studies suggest an octahedral geometry for all the complexes. The compounds were found active against B. subtilis and S. aureus and P. aeruginosa and E. coli bacteria. The Zn (II) complex showed significant anticancer activity against Squamous Cell Carcinoma cells tested by the MTT assay method. Molecular docking studies with EGFR tyrosine kinase were also carried out. All these results show that some of the synthesized compounds have remarkable antibacterial and anticancer property. 1

Introduction

A large number of macrocyclic metal complexes have been reported as antibacterial [1], anticancer [2] antifungal [3], antioxidant [4] and M.R.I contrast agents [5]. The metal ions accelerates drug action and efficiency of therapeutic compounds [6]. Tweedy’s chelation theory reported that chelation/coordination reduces the polarity of metal complexes and accelerates penetration into bacterial lipid membranes. The metal complexes

may inhibit the protein

synthesis because of interference in the cellular respiration [7] [8]. Although, Cancer is leading cause of fatality in recent years and remained as a challenge for the world [9]. The available chemotherapeutic agents have many undesirable side effects [10] so a need occur to develop safe anticancer drugs. The epidermal growth factor receptor (EGFR) tyrosine kinase is considered as an significant target for drug development because of its important roles in aberrant signaling, growth regulation and apoptosis of cancer cells [11] [12]. The EGFR inhibitors have made significant contribution in the development of anticancer agents e.g. erlotinib and gefitinib [13].

The review literatures reveal that isatin have been reported as a versatile molecule for designing various bioactive compounds like antiviral [14], antibacterial [15], antifungal [16] [17], anticonvulsant [18] and anti-HIV [19] agents. We report here the template synthesis of macrocyclic complexes of succinic acid dihydrazide and 5-chloroisatin in presence of Co(II), Ni(II), Cu(II) and Zn(II) metal ions. In vitro antibacterial and anticancer activity of complexes were evaluated. The molecular docking has been performed to study the inhibitory effect on EGFR tyrosine kinase. In excess activation of receptor tyrosine kinase (RTK) signaling pathways is strongly linked with carcinogenesis. Thus it is becoming reasonable that impaired deactivation of RTK may be a mechanism in cancer. Therefore we preferred EGFR tyrosine kinase for docking study of synthesized complexes.

2

Experimental

2.1

Materials and methods

Analytical reagent grade 5-chloroisatin, Succinic acid dihydrazide, Methanol, Ethanol, Dimethyl sulfoxide (DMSO), Dimethyl formamide (DMF), Acetonitrile, Acetic acid, Acetone, divalent transition metal salts of Sigma-Aldrich, CDH, HPLC and Fisher Scientific make were used. The C H N analysis was done with elemental analyzer Perkin Elmer, model 2400. Electronic spectra were recorded with LAMBDA 25 PerkinElmer spectrophotometer ranging from 200-1100 nm in DMSO. The infrared spectra were recorded with Thermo Scientific Nicolet iS50 FTIR Spectrometer (4000-400 cm-1) using KBr pellets and attenuated total reflection (ATR) for powder sample. The 1H NMR spectrum was recorded by Bruker spectrometer in CDCl3 at 400 MHz. The percentage of metal was determined by Atomic absorption spectrophotometer, PG instruments. Magnetic susceptibility was measured on Gouy balance. Molar Conductivity (10-

3

M) were measured on digital conductivity meter, Hach. Thermo gravimetric analysis (TGA)

was carried out with Universal TA instruments. Mass spectra were recorded with Agilent G6530AA (LC-HRMS-Q-TOF) and DART mass with JMS- T100LC, Accu TOF Mass Spectrometer, m/z ranging from 50 -1000 amu. The X-band ESR spectra were recorded by ESRJEOL Model: JES - FA200 ESR Spectrometer (free radical g = 2.00277) at room temperature.

2.2

Synthesis

In a hot stirring 20 mL methanolic solution of succinic acid dihydrazide (1.46 g, 10 mmol), a 20 mL of methanolic solution of 5-chloroisatin (1.81 g, 10 mmol) was added and refluxed for 30 min. After that 20 mL methanolic solution of metal salt (10 mmol) and 2–4 drops of glacial acetic acid were added. The mixture was refluxed for 8-10 h and filtered the precipitates. Then washed with methanol and dried in vacuum desiccators and purity was checked by TLC. Yield: 56-62%.

2.3

Antibacterial activity

Preliminary screening of the complexes against B. subtilis and S. aureus and P. aeruginosa and E. coli bacteria done by reported agar well diffusion method [20] [21] and minimum inhibitory concentration (MIC) determined by reported macrodilution tube method [22]

Preliminary screening: Mueller Hinton agar (MHA) petri plates were prepared and swabbed with 100 μL bacterial inoculum of 0.5 McFarland standard turbidity in Muller Hinton Broth (MHB). After 15 minute wells (8 mm) were created and filled with 100 μL of test compounds and ciprofloxacin dissolved

in DMSO (2 mg/mL). DMSO used as negative control. After 24 h incubation at 37 °C, Thezone of growth inhibition were measured for each compound.

MIC Determination: The complexes which exhibited remarkable antibacterial activity in preliminary screening were screened for determination of MIC against bacterial strains by macrodilution tube method using different concentrations (128, 64, 32, 16, 8, 4, 2, 1, 0.5 and 0.25μg/mL) in DMSO. Ciprofloxacin were also tested for MIC. After 24 h incubation at 37 ºC, the lowest concentration which inhibited the growth of microorganism was considered as MIC.

2.4

Anticancer activity

The synthesized complexes were tested against Squamous Cell Carcinoma (SCC4) cell line by the reported MTT assay method [23] [24]. SCC4 cells obtained from human tongue carcinoma. For the test, 100 μL (5x103 cells) of cell suspension was filled in 96 well plates and incubated for 24 h in a highly humidified atmosphere with 5% CO2 at 37°C. After that, the medium was discarded and a 200 μL of test compounds of 10 μM - 100 μM in 5% DMSO were added to wells and incubated for 24 h, 48 h and 72 h in CO2 incubator at 37 °C. Cisplatin and DMSO were also tested. Thereafter, 20 μL of 5 mg MTT /mL phosphate buffer saline solution was added and incubated for 4 h again. After incubation purple colour formazan crystals were observed. To dissolve the crystals 150 μL DMSO was added. ELISA plate reader was used to record absorbance at 570 nm wavelength. The IC50 value was calculated for each compound. Samples were analyzed in triplicate.

2.5

Molecular Docking studies

Molecular docking studies of the complexes which found highly active in anticancer study were performed by Discovery Studio 4.0, Accelrys Software, Inc. CDOCKER protocol was used to dock ligand (synthesized compound) at rigid active site

of EGFR tyrosine kinase [25]. It

implements CDOCKER algorithm and it is a grid based method [26]. The EGFR-erlotinib complex (PDB ID: 1M17) co-crystal structure with resolution of 2.6A° was downloaded from Data Bank (www.rcsb.org). The erlotinib ligand and water molecules were detached and hydrogen was added to the structure. The active site of 10.5A° radius into EGFR tyrosine kinase was defined according to PDB site record and CHARmm force field was used to prepare the protein [27]. The 3D structures of complex was prepared by ChemDraw Ultra 12.0 and minimized to RMS gradient of 0.01 using Conjugate Gradient Algorithm. The minimized structure of complex was docked at selected active site. The binding free energy was calculated for the best docked pose of complex [28].

3

Results and discussion

All synthesized complexes were coloured, soluble in DMSO and DMF, non-hygroscopic and non-electrolytic nature in DMSO (Molar Conductance, 6.48-14.27 ohm-1 cm2 mole-1) [29]. On the basis of various analytical study the synthesized complexes may be written by [M(C12H10N5O2Cl)X2] as given in Fig.1. Here, M = divalent metal ion and X = Cl–, NO3– or CH3COO– ion. The spectroscopic and magnetic studies conclude octahedral geometry of the complexes. The elemental analysis results agree well with the calculated values of elements and results are listed in Table 1.

3.1

Infrared Spectra

The IR spectrum of succinic acid dihydrazide has a pair of bands at 3260 and 3315 cm -1 corresponding to starching vibrations of ν(NH2). However, the spectra of synthesized compounds have a single medium band at 3177-3286 cm–1, it may be assign to ν(NH) stretching vibrations as shown in Fig. 2. The spectrum of dihydrazide has a strong band at 1671 cm–1 corresponding to ν(C=O) of the CONH group. But, In metal complexes this band shifted to 1618–1654 cm–1 suggesting coordination of oxygen with the metal [30]. The strong bands of ν(C=O) vibrations at1730 - 1745 cm-1 of 5-chloroisatin were absent in complexes and new strong bands were observed at 1588-1597 cm-1 assigned to stretching vibrations of azomethine ν(C=N) group [31]. The lower values of ν(C=N) vibration suggests that azomethine nitrogen atom coordinate to metal ion [32].The bands observed at 3012-3048 cm-1 , 1416-1586 cm-1 and 1022-1352 cm-1 assigned for ν(C-H), ν(C=C) and ν(C-N) vibrations respectively of 5-chloroisatin [32][33]. The complexes which contains nitrate showed three (N–O) bands at 1415–1460 cm–1 (ν5), 1302– 1318cm–1 (ν1) and 1018–1035 cm–1 (ν2). The separation of the two bands (ν 5 − ν1) suggests coordinated of nitrate groups in unidentate mode [34] [35]. The acetate containing complexes have bands at 1628–1639 cm–1 (ν1) and 1378–1394 cm–1 (ν2), It suggest that acetate group coordinates in unidentate mode [36]. The bands at 512–566 cm–1 and 462–497 cm–1 were corresponding to ν(M–O) and ν(M–N) vibrations respectively [37] [38].

3.2

1

H NMR spectra

The spectrum of Zn(II) complex exhibited a singlet peak at 2.46 ppm corresponding to -CH2 protons [39]. The multiple peaks at 7.22 – 7.94 ppm observed due to hydrogens of aromatic ring of 5-chloroisatin [40]. A singlet peak appeared at 8.83 ppm, may be assigned to –NH protons of the –CONH moiety of succinic acid dihydrazide [41]. A broad singlet peak observed at 10.89 ppm may be corresponding to –NH group of 5-chloroisatin [42].

3.3

Electronic spectra and Magnetic studies

DMSO was used to record the electronic spectra and Gouy balance for magnetic moment measurement. The ligand field parameters i.e. B′, β, and ν2/ν1 were calculated to study the nature of complex. Cobalt complexes: The Co(II) complexes absorption spectra exhibited three bands at 9542-11860 (ν1), 14987 – 17008 (ν2) and 20108 – 20434 cm-1 (ν3) corresponding to 4T1g (F) → 4T2g (F), 4T1g (F) → 4A2g (F) and 4T1g (F) → 4T1g (P) transitions. It is the characteristic of octahedral geometry [20] [43]. The complexes shown magnetic moments values µeff = 4.83–4.89 B.M. in agreement with the octahedral geometry corresponding to three unpaired electrons [20] [44]. The value of Racah parameter B’ found from 850-858 cm-1 for the complexes whereas free ion value is 971cm-1. It indicates covalence nature. The naphelauxetic parameter β value is 0.87-0.88 indicates appreciable covalent nature of bond. Furthermore, the value of the ν 2/ν1 ratio is 170-1.73 supports the octahedral geometry of the complexes. Nickel complexes: Ni(II) complexes exhibited three absorption bands at 12224–12298 cm-1(ν1), 15892–16844 cm1

(ν2), and 26936–27605 cm-1(ν3) for 3A2g (F) → 3T2g (F), 3A2g (F) → 3T1g (F) and 3A2g (F) → 3T1g

(P) transitions respectively and supports octahedral geometry [45] [46]. The magnetic moment were around µeff = 2.97-3.10 BM in agreement with the octahedral geometry with respect to two unpaired electrons [47]. The Racah parameter B’ was calculated 638-650 cm-1 whereas for free Ni(II) ion is 1041 cm-1. The Nephelauxetic parameter β is about 0.59-0.63. These indicate the covalent nature of metal ligand bond. Also the ratio of ν 2/ν1 (1.45-1.47) supports octahedral geometry [47] [48]. Copper complexes: The spectra of Cu (II) complexes show single broad band at about15504–19065 cm-1 it may be due to the mixing of all transitions like 2B1g → 2A2g (ν1), 2B1g → 2B2g (ν2), and 2B1g → 2E1g (ν3). The broadness of band may be because of Jahn Teller effect. The magnetic moment, µeff = 1.811.85 BM suggesting the distorted octahedral geometry [46] [49]. Zinc complex: The Zn (II) complex spectrum has only one band at 32720 cm-1 assignable to charge transfer from ligand to metal. Zn (II) complex is diamagnetic due to complete d-orbital therefore there is no appearance of d→d transitions and octahedral geometry is proposed supported by molar conductance results [48].

3.4

ESR Spectra of Copper complexes

The X-band spectra were recorded on a frequency of 9.4 GHz at room temperature. The values of g|| = 2.164 - 2.220 and g⊥ = 2.092 – 2.204 have been found. The values of g|| (2.164 - 2.220) > g⊥ (2.092 – 2.204)> 2.0023 point out that unpaired electron is situated in dx2-y2 ground state and it supports the distorted octahedral geometry. The values of g|| below 2.3, indicates covalent character of the metal ligand bond. The value of geometric parameter (G = 1.26-2.14) was find

out by the relationship, G = g ||-2.0023/g⊥-2.0023. By the Hathaway and Billing if G > 4, the exchange interaction is insignificant in the solid complex but when G < 4 a significant exchange interaction [49] [50].

3.5

Mass spectral studies

The obtained molecular ion peaks are mentioned in the Table 1 which supports the stoichiometric compositions of the complexes e.g. mass spectrum of Zn(C 12H10N5O2Cl)Cl2 complex exhibited M+ peak at (m/z) 426.9 a.m.u. which agree with molecular formula proposed from elemental analysis data as shown in Fig.3.

3.6

Thermal studies

Thermo-gravimetric analysis (TGA) of the complexes shows that weight loss start from 200 ºC onwards. It indicate the absence of any lattice or coordinated water and decomposed in a single stage from 200-500 ºC with the formation of respective metal oxides e.g. TGA of Co(C12H10N5O2Cl)Cl2 complex in Fig. 4 shows weight loss from 200-450 ºC and loss is about 90%. This weight loss may be due to continuous sublimation of ligand moieties. At 700 ºC an air stable compound remains which may be due to the formation of metal oxide.

3.7

Antimicrobial activity results

The complexes were found active against test microorganisms by agar well diffusion assay. The antimicrobial activity may be due to the following mechanisms: (a) By inhibition of ribonucleoside diphosphate reductase enzyme which helps in DNA synthesis; (b) By oxidative

rupture, creation of lesions in DNA strand; (c) By binding to the nitrogen base of DNA or RNA results in inhibit base replication. The compounds were screened for their MIC upto 128 µg/mL and results are given in Table 2. The results reveal that some of the metal complexes have good antibacterial property but less than ciprofloxacin. Among all the complexes against B. subtilis, the complex no. 4 & 10 were found highly active. Against S. aureus, the complex no. 8 & 10 have shown best antibacterial activity. Against P. aeruginosa the complex no. 8 was found highly active. Against E. coli complex no. 9 was moderately active. However, other complexes showed moderate to low antibacterial activity.

3.8

Anticancer activity results and discussion

In vitro MTT assay was performed to analyze the anticancer effect of complexes against SCC4 cells at different time points. The IC50 values were calculated by dose survival curves and given in Table 3. Cisplatin was used as a standard anticancer drug. Some complexes exhibited the remarkable anticancer activity against SCC4 cancer cell line with IC50 value less than 100 μM. All compounds showed time dependent effect with decrease in IC50 value at longer time period. The Zn(II) complex [Zn(C12H10N5O2Cl)Cl2] and Ni(II) complex [Ni(C12H10N5O2Cl)(OAc)2] exhibited the highest anticancer activity with IC50 values 41.4 μM and 45.4 μM respectively at 72 h.

3.9

Molecular docking study results

The compounds which have shown highest in vitro anticancer activity were docking within active site of EGFR tyrosine kinase consisting of ALA719, ASP831, GLN767, GLY772,

LEU694, LEU764, LEU768, LEU820, MET769, PHE771, PRO770, THR766 and THR830 amino acid residues. The compounds were successfully docked and studied for type of interactions with receptor protein like electrostatic interactions, hydrogen bonding, and hydrophobic bonding. The best docked pose of Zn(C12H10N5O2Cl)Cl2 complex with EGFR is shown in Fig. 5. The complex exhibited eight hydrophobic interactions with ALA719, VAL702, LYS721 and LEU820 at bond distance of 3.69A° to 5.41A° and six electrostatic interactions with LYS721, ASP831 and PHE699 amino acid residues at bond distance of 2.89A ° to 5.35A° as shown in Fig. 6. The complex showed good affinity to the EGFR receptor protein, it may be due to more hydrophobic interactions. The binding free energy of Zn complex was -402.29 kcal/mol. The obtained results show a good correlation between in vitro anticancer activity and binding free energy of Zn complex. On the bases of molecular docking results, It may be conclude that Zn complex have good anticancer potential.

Conclusions Different physico-chemical studies were used for characterization of synthesized complexes and octahedral geometry has been assigned. Two oxygen and two nitrogen are coordinated with the metal. The Cu(C12H10N5O2Cl)(NO3)2 and Zn(C12H10N5O2Cl)Cl2 complexes have highest antibacterial property among all the complexes. In vitro anticancer study shows that Ni(C12H10N5O2Cl)(OAc)2 and Zn(C12H10N5O2Cl)Cl2 complexes have remarkable anticancer property against SCC4 cell line. Furthermore, molecular docking studies have also reveal that the Zn complex have strong binding affinity with EGFR tyrosine kinase and act as an EGFR inhibitor. Thus, it supports that

Zn complex may have highest anticancer potential. In conclusion, these finding may be helpful for the design of novel antibacterial and anticancer agents. In vivo anticancer activity may be the future scope of the work.

Acknowledgements Authors acknowledge the thanks to Thermo Fisher Scientific Technical Centre- Mumbai and SAIF, IIT Bombay for spectral studies and Department of Applied Chemistry, Gautam Buddha University for laboratory facilities.

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Fig. 1: Synthesis of macrocyclic metal complexes

Fig. 2: FTIR spectrum of [Ni(C12H10N5O2Cl)(NO3)2] complex

Fig. 3: Mass Spectra of [Zn(C12H10N5O2Cl)Cl2]

Fig. 4: TGA of [Co(C12H10N5O2Cl)Cl2] complex

Fig. 5: Molecular docking of Zn(C12H10N5O2Cl)Cl2 complex with EGFR tyrosine kinase

Fig. 6: Zn(C12H10N5O2Cl)Cl2 complex Interaction with amino acid residue at active site of EGFR tyrosine kinase

Table 1: Analytical and physical data of synthesized compounds S.

Compound

N.

1

2

3

4

5

6

7

8

9

Elemental

analysis,

found

Colour

(Calculated)%

[Co(C12H10N5O2Cl)Cl2] [Co(C12H10N5O2Cl)(NO3)2]

[Co(C12H10N5O2Cl)(OAc)2] [Ni(C12H10N5O2Cl)Cl2] [Ni(C12H10N5O2Cl)(NO3)2] [Ni(C12H10N5O2Cl)(OAc)2] [Cu(C12H10N5O2Cl)Cl2] [Cu(C12H10N5O2Cl)(NO3)2] [Cu(C12H10N5O2Cl)(OAc)2]

10 [Zn(C12H10N5O2Cl)Cl2]

M.P. M+ (ºC)

Peak (m/z)

M

C

H

N

13.88

34.06

2.34

16.58

Dark

(13.98)

(34.19)

(2.39)

(16.61)

Yellow

12.21

30.28

2.10

20.48

Dark

(12.42)

(30.37)

(2.12)

(20.66)

Yellow

12.40

40.91

3.35

14.86

Dark

(12.57)

(41.00)

(3.44)

(14.94)

Brown

13.90

34.08

2.32

16.49

Lime

(13.93)

(34.21)

(2.39)

(16.62)

Yellow

12.24

30.33

2.05

20.54

Dark

(12.37)

(30.38)

(2.12)

(20.67)

Yellow

12.49

40.97

3.40

14.82

Dark

(12.53)

(41.02)

(3.44)

(14.95)

Brown

14.83

33.77

2.22

16.35

Lime

(14.91)

(33.82)

(2.37)

(16.43)

Yellow

13.18

29.98

2.08

20.30

(13.26)

(30.07)

(2.10)

(20.46)

13.31

40.52

3.34

(13.43)

(40.60)

15.16 (15.28)

267

419.9

270

473.9

266

468.0

273

420.9

279

472.9

270

467.0

276

425.9

Brown

269

477.9

14.71

Brown

280

472.0

(3.41)

(14.80)

Red

33.50

2.29

16.26

Orange

259

426.9

(33.68)

(2.35)

(16.36)

Table 2: MIC determination results of synthesized compounds against bacterial strains S.N.

Compound

MIC (µg/mL) B. subtilis

S. aureus

P. aeruginosa

E. coli

>128

>128

>128

>128

1

[Co(C12H10N5O2Cl)Cl2]

2

[Co(C12H10N5O2Cl)(NO3)2]

32

16

64

128

3

[Co(C12H10N5O2Cl)(OAc)2]

32

>128

64

128

4

[Ni(C12H10N5O2Cl)Cl2]

128

>128

128

64

5

[Ni(C12H10N5O2Cl)(NO3)2]

>128

>128

>128

>128

6

[Ni(C12H10N5O2Cl)(OAc)2]

>128

>128

>128

>128

7

[Cu(C12H10N5O2Cl)Cl2]

>128

>128

>128

>128

8

[Cu(C12H10N5O2Cl)(NO3)2]

8

8

8

128

9

[Cu(C12H10N5O2Cl)(OAc)2]

16

16

128

32

10

[Zn(C12H10N5O2Cl)Cl2]

8

8

32

64

Ciprofloxacin

5

5

5

5

Table 3: MTT Assay results of synthesized compounds against SCC4 Cell line

S.N.

Compound

IC50 Value (µM) 24 h

48 h

72 h

1

[Co(C12H10N5O2Cl)Cl2]

>100

97.2

95.5

2

[Co(C12H10N5O2Cl)(NO3)2]

>100

99.8

96.0

3

[Co(C12H10N5O2Cl)(OAc)2]

>100

>100

>100

4

[Ni(C12H10N5O2Cl)Cl2]

>100

98.0

94.4

5

[Ni(C12H10N5O2Cl)(NO3)2]

64.9

60.8

58.6

6

[Ni(C12H10N5O2Cl)(OAc)2]

50.5

47.5

45.4

7

[Cu(C12H10N5O2Cl)Cl2]

88.1

85.3

82.9

8

[Cu(C12H10N5O2Cl)(NO3)2]

>100

99.6

96.2

9

[Cu(C12H10N5O2Cl)(OAc)2]

97.6

95.0

93.6

10

[Zn(C12H10N5O2Cl)Cl2]

46.9

43.0

41.4

Cisplatin

8

6

5