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www.wjpps.com Vol 3, Issue 6, 2014.

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Muruganandam World Journal of Pharmacy and Pharmaceutical Sciences

IN VITRO ANTIBACTERIAL, ANTIFUNGAL, CYTOTOXIC AND

ANTICANCER STUDIES OF Co(II), Ni(II) AND Cu(II) METAL

COMPLEXES OF A NEW MANNICH BASE LIGAND N-

[MORPHOLINO(PHENZYL)METHYL]BENZAMIDE

L.MURUGANANDAM

Dr. Loganathan Muruganandam

Department of Chemistry, Saranathan College of Engineering, Tiruchirapalli-12

ABSTRACT

The coordination complexes of Co(II), Ni(II) and Cu(II) derived from

N-[Morpholino(phenzyl)methyl]benzamide(MBB) have been

synthesized by Mannich condensation reaction. These compounds

have been characterized by elemental analysis, IR, molar conductance,

electronic spectra, 1H & 13C-NMR, FAB-mass, ESR, magnetic

susceptibility and thermal analysis. Analytical data reveal that all the

complexes exhibited 1:1 (metal: ligand) ratio with coordination

number 4 or 6. IR data shows that the ligand coordinates with the

metal ions in a bidentate and tridentate manner through the carbonyl

oxygen, azomethine nitrogen and amido nitrogen. The Mannich base

and metal complexes are active against the six bacterial species and two fungal species.

Cytotoxic and anticancer activities of all the compounds were also studied. These studies

indicate that the activity was increased in the complexes relative to the parental ligand.

Keywords: Molecular architecture, Stereochemistry, Axial symmetry, Chelation theory.

INTRODUCTION

Metal complexes have received considerable attention for many years, due to their

interesting characteristics in the field of material science and biological systems[1].

Optoelectronic, electrical and magnetic properties of the metals and metalloids can be

tailored by reacting them with different ligands[2-4]. The Mannich bases have high affinity to

chelate with the transition metal ions: hence are attracting attention due to potential

WWOORRLLDD JJOOUURRNNAALL OOFF PPHHAARRMMAACCYY AANNDD PPHHAARRMMAACCEEUUTTIICCAALL SSCCIIEENNCCEESS SSJJIIFF IImmppaacctt FFaaccttoorr 22..778866

VVoolluummee 33,, IIssssuuee 66,, 11558855--11559988.. RReesseeaarrcchh AArrttiiccllee IISSSSNN 2278 – 4357

Article Received on 12 April 2014, Revised on 30 April 2014, Accepted on 21 May 2014

*Correspondence for Author

Dr. Loganathan

Muruganandam

Department of Chemistry,

Saranathan College of

Engineering, Tiruchirapalli


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applications[5-8] in areas viz. biology, catalysis, thermal, electrical, optical, magnetic etc.

Metal complexes are suitable to molecular materials, on the basis of electronic properties

associated with the metal center. Mannich base ligands containing O and N donor atoms play

an important role in coordination chemistry related to catalysis and enzymatic reactions,

magnetism and molecular architecture[9]. Metal complexes with Mannich base ligands

containing benzamide and its derivatives; have been extensively studied[10,11].

In this study, we report the synthesis, physicochemical characterization and biological

significance of Co(II), Ni(II) and Cu(II) complexes derived from N-[Morpholino(phenzyl)

methyl]benzamide(MBB). The reaction was carried out by Mannich condensation method.

The metal complexes formed with this new ligand are used as precursors for the synthesis of

new compounds.

MATERIALS AND METHODS

All the reagents used for synthesizing the ligand and its complexes were of A.R. grade and

the solvents used were commercial products of the highest available purity and were further

purified by distillation.

Micro elemental data were obtained with Carlo Erba 1108 elemental analyzer. Metal

contents were estimated by usual procedure[12], after digesting the complexes with

con.HNO3. Conductance data were obtained in ~10-3 M DMF solution of the complexes

using digital conductivity meter. IR spectra were recorded using Perkin Elmer FT-IR

spectrometer by using KBr pellets. Absorbance in UV-Visible region was recorded in DMF

solution using UV-Visible spectrometer. The 1H and 13C NMR of the ligand were recorded

on a Bruker instrument employing TMS as internal reference and DMSO-d6 as solvent. The

FAB mass for the ligand was carried out using Mass spectrometer. The room temperature

magnetic susceptibility measurements of the complexes were made by using a Gouy

Magnetic Balance. The antimicrobial activity was determined with the disc diffusion

method.

Antimicrobial Screening

Antibacterial and antifungal activities[13] of the ligand and six of its metal complexes were

tested in vitro against six bacterial species viz E.coli, P.aeruginosa, S.typhi, B.subtilis,

S.pyogenes, S.aureus and two fungal species A.niger and A.flavus by disc diffusion method

using agar nutrient as medium and gentamycin as control. The paper disc containing the


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compound (10, 20 and 30 µg/disc) was placed on the surface of the nutrient agar plate,

previously spread with 0.1 mL of sterilized culture of microorganism. After incubating this

at 37oC for 24 hrs, the diameter of inhibition zone around the paper disc was measured.

Anticancer Screening

In-Vitro Bioactivity Test for Immunomodulatory and Cytotoxicity Assessment

Separation of Human Peripheral Blood Mononuclear Cells

Venous blood was drawn and defibrinated in a sterile flask containing stone beads by gently

swirling the flasks. Blood was then diluted with equal volume of RPMI 1640 media and

carefully layered over histopaque in the ratio of 2:1. After centrifugation for 20 minutes, at

25˚C, the whole mononuclear cell layer seen at the interface was carefully transferred to a

tube containing RPMI 1640 medium. The cells were thoroughly mixed with the medium and

washed by centrifugation. One drop of cell suspension and one drop of tryphan blue solution

were mixed, fed into a haemocytometer. Live and dead cells were counted under phase

contrast objective. The cell concentration was adjusted to the desired number of viable

mononuclear cells/mL of RPMI 1640 medium. Care was taken to obtain a cell suspension

with 95% to 98% mononuclear cells with less than 5% to 7% contamination of erythrocytes,

granulocytes, platelets and dead cells14.

MTT Assay Method

A known concentration of the complexes was reconstituted with known volume of PBS pH-

7.2, which was the stock concentration of the complexes. This was centrifuged and the

supernatant was membrane filtered and used. From the stock, various concentrations of the

complexes were prepared and subjected for the in-vitro analysis on human PBMC. Constant

cell number was maintained. The assay was performed in a 96 well tissue culture plate using

various negative controls like plain media, complete media, vehicle, cell and solutions of

complexes. Positive controls like a known immunomodulator, PHA and a known cytotoxic

compound, LPS were also maintained15. After the addition of cell media and complex

solution, the cultures were incubated in an incubator with 95% air, 5% CO2 and humidified

atmosphere at 37˚C.

The assay was monitored after 72 hours based on spectrophotometer method. After reading

the plates, the tetrazolium compound was added to the wells and incubated. After incubation

period, the MTT was reduced by mitochondrial dehydrogenase as a result of which the

colour changed. Detergent was added to the wells to solubilize the formazan crystals. The


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absorbance was read by making use of an ELISA reader at 570 nm. The data was analyzed

by plotting cell number versus absorbance in a graph. The rate of tetrazolium reduction is

directly proportional to the rate of cell proliferation16.

Cancer Screening

Cell Lines

Raji and Jukart cell lines were used in this study. The number of cells is massively expanded

in a minimal number of passages, and the cells are cryopreserved to provide a consistent,

long-term frozen stock for future use. Cells are grown and passaged in antibiotic-free growth

medium to ensure the absence of microbial contaminants.

Preparation and Inoculation of cells

Cells are separated into single cell suspensions, and then counted using tryphan-blue

exclusion on a Hemocytometer. After counting, dilutions are done to give the appropriate

cell densities for inoculation onto the microtiter plates. Cells are inoculated in a volume of

100 µL per well at densities between 5000 and 4000 cells per well. A 100 µL aliquot of

complete medium is added to cell free wells. Prior to inclusion of cell lines in the screening

panel, their growth and compatibility were determined17.

Sample preparation

All the samples were initially solubilized in phosphate buffer saline at 1:1 ratio and were

filtered using membrane filter to avoid microbial contamination.

MTT Assay

The assay was performed in a well tissue culture plate, using various negative controls and

positive controls. After the addition of cell, media and complexes, the cultures were

incubated in an incubator with 95% air, 5% CO2 and humidified atmosphere at 37˚C for 72

hours. The assay was monitored based on spectrophotometer method. After reading the

plates, the 10 µL tetrazolium compound MTT was added to the wells and incubated. After

incubation period, the MTT was reduced by mitochondrial dehydrogenase as a result of

which the colour changed. Detergent SDS was added to the wells to solubilize the formazan

crystals. The absorbance was read by making use of an ELISA reader at 570 nm. The rate of

tetrazolium reduction is directly proportional to the rate of cell proliferation.


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Synthesis of the ligand

N-[Morpholino(phenzyl)methyl]benzamide(MBB) was synthesized by employing the

Mannich synthetic route[18]. Ethanolic solution of benzamide (12.1 g, 0.1 mol) was mixed

with benzaldehdye (10 mL, 0.1 mol) followed by morpholine (9 mL, 0.1 mol) with constant

stirring in an ice bath. After 15 days, the colourless solid obtained was washed with water

and with acetone. The compound was dried at 60oC and recrystallised from ethanol. It was

insoluble in water but completely soluble in methanol, ethanol, propanol, butanol,

chloroform, carbon tetrachloride, acetone, ether, benzene, n-hexane, dimethyl formamide etc.

Synthesis of metal complexes

The methanolic solution of each of the metal salts was added slowly with constant stirring to

the ethanolic solution of the ligand in 2:1 mol ratio at warm condition. The insoluble

complex formed in each case was filtered, washed with ethanol and methanol to remove the

unreacted metal and ligand, and then dried at 80oC.

RESULTS AND DISCUSSION

Structural Characterization of MBB

Molecular formula: C18H20N2O2, Yield: 96%, MP: 162-165oC, Mol.wt: 218, FT-IR KBr in

cm-1: 3292(NH), 1637(C=O), 3063, 3030(νCH aromatic), 2962, 2914, 2893(νCH alicyclic),

2852(νCH aliphatic), 1602(νC=C, νC-N), 1522(δCH2 morpholine ring), 1489, 1447(δNH

secondary amide), 1354, 1330, 1310(νCN mixed with δNH), 1210(νring), 1136, 1071(νCNC)

1111(C-N-C), 1024(νC-O-C as + νC-O-C sy morpholine), 1007,948(δCH+o.p.b morpholine),

915(πCH+ δCH+ o.p.b ring), 747-897(δNH wagging and twisting) , 747(δCH o.p.b benzene ring)

698(o.p.b of ring C=C-H benzene), 634(i.p.b of benzene), 409(o.p.b ring C=C). 1H NMR

(300MHz, DMSO-d6): δ 8.82(s, NH), 5.93 & 5.90(d, CH), 7.94 - 7.27(m, CH benzene ring)

3.65 (s, O(CH2)2 of morpholine), 2.51(s, N( CH2)2 of morpholine). 13C NMR (300MHz,

DMSO-d6): 167.62 (s, C=O), δ134.73-127.90(m, C phenyl ring), 66.74(s, O(CH2)2 of

morpholine), 49.33(s, N(CH2)2 of morpholine). UV-Vis(DMF): 274(n→π*), 236(π→π*).

FABMS: m/z = 296(C18H20N2O2), m/z = 120(C7H6NO+), m/z = 105(C7H5O+), m/z = 70

(C6H5+). Calculated: C 72.97%, H 6.76% and N 9.46%. Found: C 73.05%, H 6.80% and N

9.53%.

Based on the data obtained from various physical and chemical studies, the molecular

structure of MBB is shown in Fig.1.


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O N

NH

O

Fig.1: N-[Morpholino(phenyl)methyl]benzamide Characterization of CoII, NiII and CuII Complexes of MBB

All the complexes were stable and non-hygroscopic in nature. They were insoluble in

organic solvents and were soluble in DMF and DMSO solvents.

Bonding Atoms and Stereochemistry

Comparison of IR Spectra of the metal complexes with that of the ligand suggests the

coordination sites of the ligand, the mode of coordination of chloride, nitrate and sulphate

groups to the metal centre. The magnetic moment, electronic and electron paramagnetic

resonance measurements of the complexes have also been carried out. From these studies, an

attempt is made to assign the stereochemistry around the metal ions.

Microanalysis

To find out the stoichiometry[19] of the complexes, the percentage of the metal ions, anions

and CHN were determined. The CHN analyses were in good agreement with the calculated

values. From the data obtained the formation of the complexes in the molar ratio (1:1) ligand

to metal (L:M) was confirmed. This was in agreement with the ratio found, according to

elemental analysis Table(1). Table 1: Analytical and Conductance Data for CoII, NiII and CuII Complexes of MBB

Complex % C Obs

(Cal.)

% H Obs. (Cal.)

% N Obs. (Cal.)

%Metal Obs. (Cal.)

%Anion Obs. (Cal.)

ΛM ohm-1cm2

mol-1 Co(NO3)2. MBB.2H2O

35.19 (34.67)

3.32 (3.53)

4.85 (4.49)

10.54 (9.46)

20.72 (19.90) 16.90

CoSO4.MBB. H2O

36.72 (36.30)

3.66 (3.36)

5.02 (4.71)

9.45 (9.90)

15.99 (16.13) 20.46

NiCl2.MBB 40.05 (40.45)

3.31 (3.75)

4.79 (5.24)

11.43 (10.99)

12.65 (13.28) 33.32

Ni(NO3)2. MBB

37.02 (36.80)

3.26 (3.41)

5.15 (4.77)

9.48 (10.00)

20.77 (21.12) 48.11

Cu(NO3)2. MBB.2H2O

37.18 (37.63)

4.06 (3.48)

5.25 (4.88)

10.71 (11.07)

21.28 (21.60) 12.64

CuSO4.MBB. 2H2O

36.88 (37.18)

3.90 (3.44)

5.24 (4.82)

8.57 (9.16)

15.32 (15.74) 16.29


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Molar Conductance Measurements

The electrical conductance measurements of the complexes in ~10-3 M DMF solutions at

room temperature were done, in order to ascertain whether the anion was within or outside

the coordination sphere of the complex. The molar conductance values “ΛM” confirmed that

all the complexes are non-electrolytes.

IR Spectra

In the IR spectra of all the MBB complexes(Table 2), the stretching frequency, νco suffers a

negative shift of 36, 02, 13, 39, 18 and16 cm-1 suggesting the coordination of carbonyl O to

the metal atom. The νCNC band suffers a negative shift of 95, 91, 88, 60, 63 and 07 cm-1

indicating the coordination of amino N atoms. So, the ligand acts as ON donor. But in CoII

sulphato complex, in addition to the C=O and C-N-C bonds the amido NH bond is also

shifted to lower frequency(45 cm-1), which indicates the tridentate bonding nature of the

ligand[20].

The IR spectra of the CoII and CuII sulphato complexes show the presence of coordinated

sulphato group. The bands at 1136, 1020 & 964 and 1148, 995 & 962 cm-1 (SO stretching

mode, ν3), 852 and 864 cm-1(ν1), 665, 628 & 604 and 661, 638 & 617 cm-1 (OSO bending

mode, ν4) and 490 and 541 cm-1 (ν2) are due to coordinated sulphato group with bidentate

chelation[21].

CoII, NiII and CuII nitrato complexes exhibit ν5, ν1, ν2 and ν6 bands at 1385, 1309, 1016 & 912

and 1311, 1052, 1384 & 912 and 1385, 1347, 1048 and 891 cm-1 respectively, due to nitrato

group. The difference between ν5 and ν1 bands is 76, 73 & 38 cm-1, suggesting the unidentate

coordination of the nitrato group[22].

Table 2: Important IR Absorption Bands (cm-1) of MBB and of CoII, NiII and CuII Complexes

Compound νNH νC=O νCNC ν3 ν4 ν1 ν2 ν5 ν6 MBB 3292 1637 1111 - - - - - - Co(NO3)2.MBB.2H2O

3398 1601 1016 - - 1309 1016 1385 912

CoSO4. MBB.H2O

3247 1635 1020 1136,1020

665, 628, 604

864 541 - -

NiCl2.MBB 3396 1624 1023 - - - - - - Ni(NO3)2.MBB 3390 1598 1051 - - 1311 1052 1384 912 Cu(NO3)2. MBB.2H2O

3434 1619 1048 - - 1347 1048 1385 891


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CuSO4. MBB.2H2O

3486 1621 1104 1148, 995, 962

661, 638, 617

852 490 - -

The bands around 3875-3247, 1704-1621, 891-852, 692-617 and 541-462 cm-1 found in the

spectra of CoII and CuII complexes of MBB respectively indicate the presence of coordinated

water molecule[23]. The Far-IR absorption bands are observed around 589&534, 470,

403&375 and 220 cm-1 which are assignable to the M-O, M-N and M-Cl stretching modes

respectively in NiII chloro complex.

Electronic Absorption Spectra and Magnetic Moment

Based on the electronic absorption spectra and magnetic movement studies of the complexes,

the octahedral structure is assigned for nitrato and sulphato complexes CoII. The chloro and

nitrato complexes of NiII exhibit tetrahedral environment around NiII ion. The distorted

octahedral geometry is assigned for nitrato and sulphato complexs of CuII.

Table 3: Colour, Electronic Spectral Bands, Transition Assignments, Metal Environment And Magnetic Moment Values Of CoII, NiII and CuII Complexes Of MBB

Complex Colour (µeff. B.M) Environment

Absorption maxima (cm-1)

Transition Assignment

Co(NO3)2.MBB.2H2O Violet (5.48) N, 5O

6987 14823 18756 25019

4T1g(F)→4T2g(F) 4T1g(F)→4A2g(F) 4T1g(F)→4T1g(P)

CT

CoSO4.MBB.H2O Pink (5.11) N, 5O

7136 15181 19024 30448

4T1g(F)→4T2g(F) 4T1g(F)→4A2g(F) 4T1g(F)→4T1g(P)

CT

NiCl2.MBB Yellowish

green (3.86)

N, O, 2Cl

3994 8457 15082

25396,28473

3T1g(F)→3 T2g(F) 3T1g(F)→3A2g(F) 3T1g(F)→3 T2g(P)

CT

Ni(NO3)2.MBB Yellowish

green (3.52)

N, 3O

3565 7849 15012 34561

3T1g(F)→3 T2g(F) 3T1g(F)→3A2g(F) 3T1g(F)→3 T2g(P)

CT Cu(NO3)2. MBB.2H2O

Green (1.79) N, 5O 11954

29103 2Eg→2T2g

CT

CuSO4.MBB.2H2O Green (1.76) N, 5O

9116, 11785 14933

23401,32856

2B1g→2A2g 2B1g→2B2g

2Eg→2T2g(F) CT


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The X band EPR spectra(Table 4) of polycrystalline nitrato and sulphato complexes of CuII

are recorded at LNT (77 K). They show EPR spectra of axial symmetry type indicating

distorted octahedral geometry around copper centre.

Table 4: EPR Spectral Parameters Of CuII Complexes

Complex g║ g┴ gav G Cu(NO3)2.MBB.2H2O 2.14 2.02 2.08 7.78 CuSO4.MBB.2H2O 2.07 2.01 2.03 8.79

Based on the above results, hexa-coordinated geometries are assigned to nitrato and sulphato

complexes of CoII and CuII. NiII chloro and nitrato complexes have tetrahedral structure. The

structures proposed, tentatively are in Fig.2, 3, 4, 5, 6 &7.

Co

ONO 2

OH2

NO

NH

O

H2O

O2NO

CoO

OS O

O

NO

NH

O

H2O

Fig.2: Co(NO3)2.MBB.2H2O Fig.3: CoSO4.MBB.H2O

Ni

ClCl

NO

NH

O

Ni

ONO2

NO

NH

O

O2NO

Fig.4: NiCl2.MBB Fig.5: Ni(NO3)2.MBB

Cu

ONO 2

NO

NH

O

O2NO

H2O OH2

Cu

OO

S

OO

OH 2

NO

NH

O

H 2O

Fig.6: Cu(NO3)2.MBB.2H2O Fig.7: CuSO4.MBB.2H2O


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Biological Studies

Antibacterial Activity

The results of the percentage of inhibition of the bacterial growth are presented in Table 5. In

all the seven compounds tested for antibacterial assay, it was found that activity increases

with an increase in the concentration of the test compound. The copper(II) nitrato complex

was found to be the most active against six bacterial species among the complexes screened.

This pronounced activity may be explained as follows. The lipids and polysaccharides are

important constituents of the cell wall and membrane, which are very much preferred for the

metal ion interaction[24]. The cell also contains many aminophosphates and carbonyl and

cysteinyl ligands, which maintain the integrity of the membrane by acting as a diffusion

barrier. They also provide suitable sites for binding. Since the nitrato complex has two labile

water molecules and is coordinately unsaturated, the metal center in this complex can

exchange water molecules for biological binding sites. As a result of chelation, the polarity

of the metal ion is reduced and the increased lipophilic character favors the interaction of

metal complexes of the cell, resulting in interference with the normal cell processes[25].

Table 5: Antibacterial Activity Of Ligand And Its Complexes

Compound E. coli P. aer. S. typhi. B. sub. S. pyo. S. aur. Conc. (µg/disc) 10 20 30 10 20 30 10 20 30 10 20 30 10 20 30 10 20 30 Control 12 15 20 10 13 18 14 17 22 11 14 18 10 12 16 12 17 20 MBB 18 21 24 17 22 27 20 28 30 23 26 32 18 24 29 25 28 35 Co(NO3)2.MBB.2H2O 22 29 32 23 26 32 21 26 32 20 29 31 20 24 31 26 28 34 CoSO4. MBB.H2O 21 25 35 24 25 35 22 28 36 20 27 35 22 26 33 23 27 36 NiCl2.MBB 25 27 36 22 26 39 25 32 40 21 30 39 25 28 35 23 29 39 Ni(NO3)2.MBB 26 32 45 24 28 40 25 34 44 23 31 42 28 32 40 26 32 41 Cu(NO3)2. MBB.2H2O 31 35 47 34 39 52 35 43 58 30 37 50 36 47 59 33 46 58 CuSO4. MBB.2H2O 29 33 45 30 36 46 31 38 49 28 36 47 32 44 52 30 41 53

Antifungal Activity

It is interesting to note that the fungitoxicity of free ligand is less severe than that of metal

chelates (Table 6). Among the complexes screened, the order of activity was found to be

Cu(II) complexes > Ni(II) complexes > Co(II) complexes > MBB. A possible mechanism of

toxicity may be speculated in the light of chelation theory[26]. Chelation considerably reduces

the polarity of the metal ion mainly due to partial sharing of its positive charge with donor

groups and possible π-delocalization over the chelate ring. This increases the lipophilic

character of the neutral chelate, which favors its permeation through lipoid layers of fungus


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membranes. Furthermore, the mechanism of action of the test compounds may involve

formation of a hydrogen bond through the uncoordinated O, S, and N hetero atoms with the

active centers of the cell constituents. This results in interference with the normal cell

process. The presence of lipophilic and polar substituents like C=O, C=S, SH, and NH are

expected to enhance the fungitoxicity. These compounds have a greater chance of interaction

with either the nucleotide bases or biologically essential metal ions present in the biosystem.

They interact with coordinatively unsaturated metal present in the metal complexes, to

achieve a higher coordination number, by reacting with some of the enzymatic functional

groups[27].

Table 6: Antifungal Activity Of Ligand And Its Complexes

Compound A. niger A. flavus Conc. (µg/disc) 10 20 30 10 20 30

Control 11 14 17 13 18 21 MBB 14 18 25 15 23 26 Co(NO3)2.MBB.2H2O 17 22 29 17 25 29 CoSO4. MBB.H2O 20 26 32 18 23 31 NiCl2.MBB 19 28 35 21 27 34 Ni(NO3)2.MBB 23 30 38 22 28 39 Cu(NO3)2. MBB.2H2O 30 37 49 31 42 55 CuSO4. MBB.2H2O 26 31 40 27 34 47

Anticancer studies

The results of cytotoxic studies of MBB and its metal complexes of CoII, NiII and CuII

against peripheral blood mononuclear cells reveal that the cytotoxicity increases as the

concentration of test compound increases[28]. A very few metal complexes are found to be

more active than the ligand. The ligand acts as very good immunopotentiator. When

complexed with metal salts, the immunopotentiating property increases(Table 7).

Table 7: in-vitro bioactivity test for immunomodulatory and cytotoxicity assessment

(MTT method)

Compound Dye Exclusion MTT MBB Non-toxic Immunopotentiator at 100 ng Co(NO3)2.MBB.2H2O Non-toxic Cytotoxic CoSO4. MBB.H2O Non-toxic Immunopotentiator NiCl2.MBB Non-toxic Non-toxic Ni(NO3)2.MBB Non-toxic Cytotoxic Cu(NO3)2. MBB.2H2O Non-toxic Non-toxic up to 200ng

Cytotoxic at 400 ng CuSO4. MBB.2H2O Non-toxic Cytotoxic


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When the compounds were analyzed for anticancer activity, the results (Table 8) revealed

that CoSO4 have up regulated[29] Raji and Jukart cell line at 50 ng. MBB was found to be non

toxic against both the cell lines tested. All the remaining complexes were down regulated for

Raji and Jukart cell lines at 50ng. In the present study, it appears that there is a relationship

between the lability of M-O and M-N linkages and the activity as discerned from the

thermodynamic and kinetic stability. Based on the above preliminary studies, the following

possible explanations are drawn

1. Compound may affect the specificity of DNA by altering the hydrogen bonding

interactions[30] by accidental incorporation of antimetabolite in DNA in the place of the

essential metabolite(thymine) and slow down the rapid multiplication.

2. The metal ion may act as an intermediate to keep the protein bound to RNA and retain the

configuration or may inhibit the protein and nucleic acid synthesis[31]. So, metal chelation

augments the activity of the drug.

3. The lability of the M-O and M-N bonds and thermodynamic stability appear to influence

the activity of metal complexes. The activity is proportional to the lability of the metal-

donor bond.

Table 8: Consolidated Results Of The Compounds Against Cancer Cell Lines

Compound Cell Lines

Raji Jukart

MBB Non toxic Non toxic

Co(NO3)2.MBB.2H2O Down regulation at 50 ng Down regulation at 50 ng

CoSO4. MBB.H2O Up regulation at 50 ng Up regulation at 50 ng

NiCl2.MBB Down regulation at 50 ng Down regulation at 50 ng

Ni(NO3)2.MBB Down regulation at 50 ng Down regulation at 50 ng

Cu(NO3)2.

MBB.2H2O Down regulation at 25 ng Down regulation at 50 ng

CuSO4. MBB.2H2O Down regulation at 50 ng Down regulation at 50 ng

In general, the intake of a drug depends on the balance between hydrophilic and lipophilic

properties and the solubility which are substituent dependent. Metal coordination increases

the lipophilicity of a drug and this may be the reason for the enhanced activity upon


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complexation. Hydrogen bonding and the antimetabolite action of the compound may be an

important factor in anticancer mechanism[32].

CONCLUSION

A new Mannich base ligand, N-[Morpholino(phenzyl)methyl]benzamide(MBB) and its

complexes of CoII, NiII and CuII were synthesized and characterized. Based on the spectral

data, the ligand acted as bidentate and tridentates to the metal ion. Tetrahedral geometry was

assigned to the NiII chloro and nitrato complexes. The remaining complexes were assigned

distorted octahedral geometry. The ligand and the complexes were screened for their

antibiological activities. All the antimicrobial studies showed that the CuII nitrato complex

was more active than the rest. ACKNOWLEDGEMENT

The author is thankful to the Management, Secretary, Principal and Dean(R&D), Saranathan

College of Engineering, Tiruchirapalli for providing facilities and motivating him with their

constant encouragement. The research work was carried out at NIT, Tiruchirapalli. The

author is grateful to Dr.T.Thirunalasundari, Department of Biotechnology, Bharathidasan

University, Tiruchirapalli, for helping them to carry out antimicrobial studies at her

laboratory. REFERENCES

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