1

CHAPTER 1
Thiosemicarbazones-Biologicallyactive materials, Analytical reagents,
Coordinating agents,structure and bonding.
1.1.Introduction:
Coordination chemistry is exactly, the chemistry of metal atoms "coordinated" by
moleculesor atoms which has always been a challenging task to the inorganic chemist.
In the beginning stage of chemistry as a separate branch of study, coordination
compounds look unusual (hence the name complexes) to hold the usual rules of
valence. But now-a-days, this compromises a large body of current inorganic
research. According to survey of various articles in recent issues of journal “Inorganic
Chemistry”indicates that perhaps 70% could be interested to deal with coordination
compounds. The usual bonding theories can be extended to accommodate these
compounds whichstill provide many challenging problems that are to be resolved. But
in synthetic work, this continues to facea challenge in the laboratory. Interdisciplinary
research work has become a mark of modern inorganic chemistry. Many chemists
today are using the concepts and methods of modern coordination chemistry to solve
the problems in research areas such as catalysis, energy conversion,materials
chemistry and biochemistryetc. The fast progressing field of bioinorganic chemistry
primarily focused on the presence of coordination compounds in many living systems.
Chemical reactions were known to man ever since chemistry had been recognizedas
specifically isolated branch of study. It was found that substances would change their
properties under specific external conditions, and this observation is a characteristic of
various chemical reactions involved. For example, the ancient Egyptians discovered
that if malachite, a green ore, was burnt with charcoal, a red metal forms which was
called as copper. The development of modern medicinal inorganic chemistry has
excited by the discovery of cis-dichlorodiamine platinum (II) [cisplatin] and its
following usage as a drug in treatment of several human tumors[1-3], has been made
easier bythe inorganic chemist's intensive knowledge on coordination and redox
properties of metal ions. The positively charged metal centers, bind to negatively
charged biomolecules and the constituents of proteins and nucleicacids offer as
excellent ligands for binding to these metal ions. Hence the metal complexes have
excellent potential in pharmaceutical usage.
 2

Thiosemicarbazones (hydrazinecarbothioamides) are a family of compounds having

high biological activity. They are very good ligands. The biological activity of these
ligandsis related to their ability to coordinate to metal centers in enzymes. An
interesting feature of pharmaceutically promising thiosemicarbazone derivatives is
that,these derivatives possess additional functional group that are not coordinated to
their "primary" metal ion, there by suggesting that the biological activity may also
depend on the non-coordinating groups. This class of compounds isin general
prepared by condensation of carbonyl group (ketone or aldehyde) with TSC or with
N(4) substitution. Based upon the nature of the ketone or aldehyde the resulting
thiosemicarbazone may differ. The following strategy may be involved to evaluate the
biological activity of TSCs.
a.Thesulphur atom of the thiocarbanyl group may be replaced by imine, oxime,
selenium or oxygen.
b. The sulphur center may be modified by alkylation.
c. The heterocyclic ring is added to TSC moiety point of attachment.
d. The terminalN(4) position, undergoing various substitutions.
e. By variation inthe nature of condensing aldehyde or ketone.
The metal complexes of TSCs have drawn attention as improved drugs because of the
following advantages:
1. Since the breakdown of metal complexes results into metal ion, which interacts
with the organism, the long term side effects of Therapeutic agents can be avoided.
2. The role of metal complexes to act as a vehicle for the ligand activation, which is
the principal cytotoxic agent.
3. Reduction of drug resistance by several magnitudes is due to complexation with
metal ion .
4. Biologically essential elements such as Cu, Fe and Zn forms large number of such
complexes.
1.2.Thiosmicarbazones in the biological field - a survey:
Nearly 50 years of search out has been done for thiosemicarbazones and their metal
complexes, because of their wide range of biological activity, it can be used in various
drugs for various diseases[4-6]. Thiosemicarbazones and their metal complexes have
been studied in various field such as anticancer[7], antitumour[8], antifungal[9],
 3

antibacterial [10], antimalarial[11], antifilarial[12], antiviral[13] and anti-HIV[14]

activities.
The delivery of radioactive isotopes of Copper tumour or leucocytes by hypoxic
selectivity of Copper bis (Thiosemicarbazones) in the form of vehicles has drawn
much attention recently[15-17]. The substituent’s on the back bone of Carbon
strongly affects the hypoxic selectivity. Previous studies say that the biological
properties of thiosemicarbazones and their metal complexes is due to biologically
active thiosemicarbazone molecule in a planar structure and presence of a pyridine
ring or NNS tridentate system[18].
But recent studies say that biological activity majorly due to parent aldehyde or
ketone[19-20], while the presence of a bulky group at the terminal Nitrogen
remarkably increases the biological activity[21]. Therefore the biological activity can
be greatly increases by an additional potential bonding site together with the presence
of bulky group at the N(4) position of thiosemicarbazone[22-24].
It is also believed that the biological activity is due to the ability of thiosemicarbazone
molecule to chelate with traces of metals in the biological system. Some of the side
effects may be reduced by coordination, the lipophilicity which controls the rate of
entry into cell, is modified[25-26]. Thiosemicarbazones may block DNA synthesis in
mammalian cells by inhibiting the enzyme, ribo-nucleosidediphosphatereductase
either bychelation with an iron ion required by the enzyme or because a preformed
metal chelate of the inhibitor interacts with the target enzyme [27-28]. The study also
show the ability of thiosemicarbazones to severe the DNA strands [29]. A major
clinical challenge in treatment of cancer with anticancer drugs is that fewtumors cells
develop a particular phenotype, known as multi drug resistance (MDR), which
transform or convert these cells resistant to other classes of anticancer agents to which
the tumor cells have not been treated earlier[30].Synthesis and characterization of a
palladium complex of phenathrenequinonethiosemicarbazone and evaluation of
itsantiproliferative properties on breast cancer cells and normal cells have been
described [31].
The studies reveal that the complex is a potent antineoplastic agent that has selective
activity against tumour cells and is much effective against drug resistant breast cancer
cells. In recent years there is much research going on biological and medicinal
 4

properties of transition metal complexes of thiosemicarbazones. Recently a square

planar complexes having a general formula [M(NNS)CI] (M = Pd(II), Pt(II); NNS =
anionic forms of the 6-methyl-2-tonnylpyridine Schiff bases of S-methyl and S-
benzyldithiocarbazates) have been prepared. Both the Schiff bases exhibit strong
cytotoxicity against the human ovarian cancer (Caov-3) cell lines.The S-methyl
derivative is two times more active than the S-benzyl derivative[32]. Palladium(II)
and platinum(II) complexes of 5-chloro-I,3-dihydro-3-[2-(phenyl)ethylidene]-2H-
indol-2-one-hydrazine carbothioamide have been synthesized and screened for their
antimicrobial activity against the fungi Macrophominaphaseolina and
Fusariumoxysporumby agar plate technique.It has been proved that these compounds
showing antimicrobial properties and metal chelates exhibit greater or high inhibitory
effects than theparent ligands. The biological potency of these complexes can be
increased by increasing the lipophilic character[33].
1.3.Thiosemicarbazones in the analytical field:
Thiosemicarbazones have immense applications in analytical field also. These
complexes produce highly colored complexes with metal ions. Thiosemicarbazones
have been proposed as analytical reagents that can be used in sensitive and selective
determinations of metal ions [34-35]. Ferrocene derivatives containing
thiosemicarbazone side chain have been investigated by position annihilation lifetime
(PAL) and cyclic voltammetry measurements. In molecular solids,Positrons can form
the positron-electron pair. Positron-electron pairis of great interest because of its
formation and also its lifetime depends upon the physical and chemical properties of
the solid. It has been proved that the electron capture & redox processes have taken
place on the Fe atom [36],Co(Il), Cu(II) and Fe(lI) in pharmaceutical preparations
could be determined using solvent extraction and pre-column derivatization with 2-
acetylpyridine-4-phenyl-3-thiosemicarbazone as complexing reagent [37].
1.4.Thiosemicarbazones - the coordinating agents:
Thiosemicarbazones are derivatives of thioureaprepared or synthesized by
condensation of N(4)-substituted thiosemicarbazide or thiosemicarbazide with a
suitable ketone or aldehyde. These thiosemicarbazones are shown by the general
formula (I) and when N(4) is substituted with a formula(II).The numbering scheme as
shown in the figure is in accordance with IUPAC system. However, it should be noted
 5

that the numbering schemes for crystal structures are in accordance with the atoms
present. In formula (II) R3 and R4 can be aryl are alkyl groups or a part of a cyclic
system. According to IUPAC recommendations for naming for naming organic
compounds,derivatives of thioureacan be shown by general formula R1R2C=N-NH-
CS-NR3R4, may be named by adding the class name of condensed ketone or aldehyde
[38,39].

Fig: 1.1 Generalstructures of thiosemicarbazones.

Thiosemicarbazoneshas a variety of different coordination modes. In such cases, they
coordinate as bidentate ligands via azomethine nitrogen and thione/thiolato sulfur.But
when additional coordination functionality is present in the proximity of the donating
centers,then the ligands will coordinate in a tridentate manner. This can be achieved
either by neutral molecules or by the monobasic anion upon loss of hydrogen. In solid
statethione form predominates whilesolutions of thiosemicarbazone molecules show a
mixture of both tautomers.Based upon the preparative conditions,the metalcomplexes
may be anionic,cationic or neutral.Hence therehave been many cases where the
anionic and neutral forms of the ligand are involved in coordination[40]. The various
coordination geometries encountered in this study may be discussed in the following
chapters.
The ligands are linked or bonded to the central metal ion through their donor atoms.
The metallic atomsto which the ligands are linked through coordinating bonds is
called the central metallic atom. The oxidation state of metallic atom may be zero,
negative or positive. Based on the number of donor atoms contained, ligands may be
classified as uni,di,tri or quadridentate.
1.5.Coordination number of central metal atom / ion:
Coordination number of the central metal atom in a given complex is equal to the total
number of donor atoms,which are actuallylinked with the central metal atom. In case
of complex compounds which contain only monodentate ligands, thecoordination
 6

number of the central metallic atom is equal to the number of mono dentate ligands
co-ordinated to the metal atom. But this rule is not applicable for the complexes
containing polydentate ligands. Coordination number of metallic atom indicates the
geometry of the complex compound. Thus for coordination numbers 2,3,4,5&6 the
geometry of the complex compounds formed are linear,trigonal,planar,tetrahedral and
octahedral respectively. Hence coordination number is that which gives us
information about the way in which the ligands are bonded around the central metal
atom[41].
1.6.Structure, bonding and stereochemistry:
In general TSCs exist in the thione form in the solid state, but in solution it tends to
exist as an equilibrium mixture. Thiol and thione forms as follows

NR1R2

H NR1R2 N
R1
N N SH
R1

N S
R2

R2

ThioneThiol
Fig 1.2 Structures of thione and thiole forms of thiosemicarbazones.
Itis assumed that the TSC generally exist in the E form (trans).In such cases the
compound may act as a unidentate ligand,by bonding through sulfur atom only[42].If
the sulfur center is substituted, the bonding may takes place by hydrazine nitrogen and
the amide nitrogen.
Generally, the coordination takes place by the thiovsthione sulfur and thehydrazine
nitrogen. But when additional binding sites are present adjacent to theS-N donor
system, the ligand will act as a tridentate species. In majority of cases the third
coordinating species will be in form of ketonicor aldehydicmoiety[45]. The
availability of the third coordinating site may lead to the formation of polymeric
complexes[46] or could change the oxidation state of metal or may bring changes in
the coordination environment of the metal ion. The changes taking place may affect
the biological properties of the compound. The alkylation of the thiocarbonyl sulfur
induces a complexation through terminal amido group and hence results for mono
 7

acidic character of the ligand[47].Such ligands may sometimes condense with a

second aldehyde or ketone to result quadridentate ligands.
The stereochemistry of the TSC ligands with the transition metal ions essentially
depends on the presence of additional coordination sites in the ligand moiety and also
on the charge of the ligand. This in turn is effected by the thiol-thione equilibrium in
the reaction medium. The equilibrium and the nature of complexationof thiol-
thionedepend on the pH of the medium/solvent(s) used. The TSCs Generally act as
neutral bidentate ligandswhich results into formation of octahedral and square planar
geometries. TSC derivatives show increased biological activity as antitumour,
antimalarial agents[48].
TSCs can bring about changesin bonding and stereochemistry of the compounds,
which may decide the mechanism of the compound inbiological systems.
HS NH2 HS NH2

N N
N N

R2 R2
R1 R1

Z(CIS)FORM E(TRANS) FORM

Fig: 1.3 Tautomerism in thiosemicarbazones
But the recent studies shows that the stereochemistry of the ligand is depends upon
the steric effects of the various substituents present in the TSC moiety [51]. While
coordinating to the metal ion, the compound is always found to be in Z-form.This
phenomenon is due to the "chelate effect" increased stability due to better electron
delocalisation in a chelated ring system- resulting from the coordination with metal
center.
1.7.Important role of metals:
Zinc was discovered centuries ago in the form of metallic which are used for making
brass and zinc. Zinc compounds were used for healing wounds and sore eyes. Zinc
ores are widespread geologically and geographically. Zinc is found abundantly in the
earth's crust which is nearly of about 132 ppm by weight. In general zinc ores are
found in combination with copper, lead, silver, gold as well as other metals. The
mined ore is rarely good enough to be used directly by the smelters which should be
concentrated. Generally Zinc ores contain 3% to 10% zinc and zinc concentrates
 8

usually contain55% zinc. Zinc is one of the essential trace element, found in high
concentration in the human red blood cells as enzymecarbonic anhydrase, which helps
to undergo many reactions like carbon dioxide metabolism. Zinc plays an important
role in biological functions such as disease, wound healing, resistance, digestion,
physical growth, reproduction, diabetes control, taste, smell [1]etc. Every cell in the
human body requires zinc to get multiplied and more than 300 enzymes require zinc
for proper functioning. According to "World Health Report 2002" estimated that one-
third (33%) of the world's population is at risk of inadequate zinc intake. The effects
of zinc deficiency may be severe, such as impaired neuropsychological functions,
growth retardation and stunting, immune disorders, impaired reproduction, lethargy,
dermatitis, loss of hair and loss of appetite [55-56].
Zinc has d10s2 electronic configuration and generally forms M2+ ions. Many of their
compounds are covalent in nature. Due to stability of the filled d orbital, this element
shows characteristic properties of transition metals despite its position in the d-block
of the periodic table. Zincalso resembles other transition metals information of stable
complexes with N,O and S-donor ligands like halide, cyanide etc. This shows the high
flexibility in the structure, co-ordination mode and coordination number of the
complexes are produced. Among these complexes, some has special attention as
model compounds for active sites of zinc-containing enzymes [57],and their functions
strongly depend upon the nature of coordination around the zinc ion. For creating
functional zinc complexes, it is very important to consider the relationship of the
coordination characteristics particular to zinc ion. Biological activity of
thiosemicarbazones and semicarbazones increases on complexation with transition
metals[58] and higher activity can be obtained with substitution at N4-
position[59].Tetrahedral and octahedral Zn(II) complexes of thiosemicarbazide and
TSC of bidentate ligands have been reported[60]. On the other hand, ligands, such as
ethylacetoacetatesemicarbazone and TSC acting astridentate in zinc complexes [61],
in which the third coordinating center is provided by the carbonyl group. This chapter
deals with the preparation and characterization of eight Zn(II) complexes of N4-
substituted thiosemicarbazones.
Cadmium is one of the relatively abundant available elements which were invented in
1817 by FredrichStromeyer. It is found as an impurity in zinc carbonate. Cadmium
 9

was named after the Latin word for calamine, since the metal was found in zinc
compound.Cadmium containing ores are very rarely found and thus to occur in small
quantities [62].Gadreenockite (CdS), the only cadmium mineral is combined with
sphalerite (ZnS).As a result cadmium is obtained as a byproduct from mining,
smelting and refining sulfide ores of zinc and to a lesser degree for lead and copper. A
very small amount of Cd (i.e) about 10% is obtained from secondary sources, mainly
from dust generated by recycling iron and steel scrap. It’s Production in United States
started in 1907, but it was not used until after World War I. The Only place where
metalliccadmium can be observed is the Vilyuy River basin in Siberia [63]. An
important commercial use of cadmium is that, it is used as an electrodeposited coating
on iron and steel for preventing from corrosion. The second-largest application is
Nickelcadmium batteries, which pigment and chemical uses third. A very small
amount of cadmium are used in low-melting-point alloys,similar to Wood metal, in
automatic fire sprinklers, in brazing alloys, solders, and bearings. Compounds of
Cadmium are also used as stabilizers in plastics and also in the production of
cadmium phosphors. Cadmium isotope 113 has a great neutron-absorbing capacity,
hence usedfor nuclear reactors. Cadmium has d10s2 electronic configuration and it
forms M2+ ions and most of its compounds are covalent in nature. The most stable
form of cadmium is Cd(II).Cd(II) complexes shows tetrahedral, square pyramidal,
trigonalbipyramidal and octahedral geometries. There is substantial interest in the
coordination chemistry of cadmium complexes due to toxic environmental effect of
cadmium. The mobilization and immobilization of Cd in the environment, in
organisms, and in technical processes have been shown to depend significantly on the
complexation of the centrlmetal by chelating with nitrogen donor ligands[64].
Cadmium forms complexes with carboxy and acetate ligands as well to result both
neutral and charged compounds. The possibility of forming geometries with higher
coordination numbers has resulted in the unusual coordination geometries about the
metal atom and the formation of polymeric species [65].The most stable Cd(II)
complexes forms with soft donor atoms (S>>N >O). The stability of the complexes
increases with the increase in number of coordination groups contributed by the
ligand. It has been noticed that some cadmium complexes of TSCs shows quite large
SHG efficiency [66].
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Copper is one of the important and most abundantlyavailable element in the earth’s
crust. It exists in two oxidation states i.e, Cu + and Cu +2
. Among this the most
common oxidation state of Cu is +2 and copper(II)complexes have been widely
studied. These complexes shows tetrahedral,octahedral, square planar and
trigonalbipyramidal geometries [67]. The role of copper in organic reactions depends
upon its oxidation states. Copper metal is widely used in electrical industry due to its
high conductivity and it is also used for water pipes because of its inert property.
Copper catalyses redox reactions arebused in biological systems primarily in the
reduction of oxygen to water [68]. Due to the presence of unpaired e-, all the
copper(II) complexes are paramagnetic in nature. More interest in Schiff base
compounds containing TSCs and their transition metal complexes haveimproved in
the area of chemistryand biology due to biological activity[69-70]. But semicarbazone
analogs got much less attention. However, they possess versatile structural features
[71]. Metal complexes containing Cu(II) and iron(II) are more active than the
uncoordinated TSCsand this increase in biological activity [72]. Cu (II) complexes are
more interesting due to their roles in biological and medicinal properties.
Nickel exists in three oxidation states such asNi(I), Ni(II) or Ni(III) in biological
systems as an active sites of certain hydrogenases and dehydrogenases [73]. Hence
nickel species are of much interest to inorganic biochemists. Nickle is predominantly
divalent and ionic in simple compounds.Nickel shows4, 5 and 6 coordinate complexes
such as square planar, tetrahedral, trigonalbipyramidal, square pyramidal and
octahedral geometries. The coordination compounds of nickel are also studied for
their magnetic property. In last few years, Ni(II) complexes containing sulfur donors
has got most considerable attention due to sulfur rich coordination environment in
biological nickel centers such as at the active sites of certain ureases, methyl-S-
coenzyme-M-methyl reductase, hydrogenases and can play an active role in
mutagenicity of nickel compounds. According to the studylabile four coordinated
Ni(II) complexes with tridentate TSC and semicarbazone ligands shows antibacterial
activities, where as six coordinated Ni(II) complexes with TSC and semicarbazone
ligands exhibit any biologicalactivity against the microorganism test[74].
The synthesis and reactivity of cobalt complexes of Schiff base ligands have played a
very important role in coordination chemistry[75-76]. Coshows+2 and +3 oxidation
 11

states and salts of Co(II) are more stablesince they are difficult to oxidized to Co(III)
state.However, in basic solutions, oxidation of Co(II) to Co(III) takes place easily.
The overall formation constant is greater for higher oxidation state and thus the
complexation makes it difficult to reduce. However, In spite of stabilization of Co(III)
by complexation, high spin 6 coordinate, high/low 5 spin coordinate and 4 coordinate
complexes of Co(II) are mostly reported. It has been noticed that Co(II) forms
tetrahedral complexes than any other transition metal ion except Zn(II) because of its
d7 configuration.The cobalt complexes having tetradentate Schiff base ligands
arewidely used to mimic cobalamin (B12) coenzymes [77-80], oxygen activators[81-
84], dioxygen carriers and also in enantioselective reduction[85].60 Radioactive
isotope of cobalt is used in radiotherapy. Co compounds are in general used in the
production ofvarnishes, inks and pigments. Co acts as ferromagnetic and the Curie
temperature observed is 1388 K with1.6~1.7 Bohr Magneton/atom. Cobalt oxides,
CoO and Co3O4 are anti ferromagnetic in nature at low temperatures.
Manganese is a naturally occurred metal, found in many rocks. Mn is widely used in
steel production to improve its strength, stiffness and hardness. It is also used as an
additive in gasoline to improve octane rating. Manganese is naturally present in the
environment. Mn is required for normal physiologic functioning in humans and
animals. Chronic exposure by inhalation affects central nervous system (CNS). Visual
reaction time, eye-hand coordination and hand steadiness were affected in
chronically-exposed workers. A syndrome called manganism would result from
chronic exposure. This is characterized by feelings of weakness and laziness, tremors,
a mask-like face and psychological disturbances. Impotence and loss of libido has
been noticed in male workers afflicted with manganism.
Manganese is also an essential trace element, forming a number of active sites for
metalloprotiens. In metalloprotiens, Mn can exist in any of the five oxidation states or
in mixed valence states [86]. In photosystem II (PSII) it is found astetranuclear
manganese complex[86]. Manganese coordination compounds have improving as
homogeneous catalysts in oxidation reactions. Manganese has an important role in
many enzymatic systems such as peroxidase, peroxide dimutase, dioxygenase in
which mononuclear manganese active sites are seen[87]. Manganese metal complexes
play an important role starting from bioinorganic chemistry to solid state physics [88-
 12

89]. Manganese complexes are also studied for their magnetic property. The
chemistry of manganese in various oxidation states in various combination of nitrogen
and oxygen donor environment is currently witnessing intense activity [90-91]. The
common oxidation states of Mnare +2, +3, +4, +6 and +7.The most common and
stable oxidation state exhibited by manganese is +2and most of these complexes are
high spin paramagnetic d5 systems and shows colored.
1.8. Objectives and Scope of the present work
As discussed previously, transition metal complexes of thisemicarbazones possess
a large spectrum of properties. The significance of thiosemicarbazones and their metal
complexes, as part of their diverse chemical and structural characteristics, stems from
not only their potential but also their proved application as biologically active
molecules have encouraged as to explore their coordination chemistry further. The
work embodied in this thesis mainly aims at weaving a network about their interesting
spectroscopic, antimicrobial properties. Several new complexes have been
synthesized and characterized using elemental analysis, molar conductance, magnetic
measurements, IR and electronic spectral studies. The present study mainly aims at:-
1. To synthesize and physico-chemically characterize the following
thiosemicarbazone ligands without substitution at the N(4)-position.

a) Acetylcyclohexanethiosemicarbazone (HL1=ACTSC)
S

N
N NH2
H

b) 3-Methylbutanalthiosemicarbazone (HL2=MBTSC)

H
H2 N N
N

S
 13

c) 4-Bromo-2-thiophenecarboxaldehydethiosemicarbazone (HL3=BTTSC)
S S
Br N
N NH2
H
2. To synthesize some metal complexes of above ligands.
3. To investigate the spectral characteristics of these complexes.
4. To study the biological activities of the ligands and the complexes