INTRODUCTION

This project has been aimed at the synthesis of some novel heterocyclic
compounds like Schiff bases and their cyclisation to produce (Azetidinones) Betalactam derivatives of biological significance.

1.1

General Introduction of Schiff bases

A Schiff base is a nitrogen analog of an aldehyde or ketone in which the C=O

group is replaced by C=N-R group. It is usually formed by condensation of an

aldehyde or ketone with a primary amine according to the following scheme:

Where R, may be an alkyl or an aryl group. Schiff bases that contain aryl
substituents are substantially more stable and more readily synthesized, while those
which contain alkyl substituents are relatively unstable. Schiff bases of aliphatic
aldehydes are relatively unstable and readily polymerizable1,2 while those of aromatic
aldehydes having effective conjugation are more stable3-6.
The formation of a Schiff base from an aldehydes or ketones is a reversible
reaction and generally takes place under acid or base catalysis, or upon heating.

The formation is generally driven to the completion by separation of the

product or removal of water, or both. Many Schiff bases can be hydrolyzed back to
their aldehydes or ketones and amines by aqueous acid or base.
The mechanism of Schiff base formation is another variation on the theme of
neucleophilic addition to the carbonyl group. In this case, the neucleophile is the
amine. In the first part of the mechanism, the amine reacts with the aldehyde or
ketone to give an unstable addition compound called carbinolamine.The
carbinolamine loses water by either acid or base catalyzed pathways. Since the
carbinolamine is an alcohol, it undergoes acid catalyzed dehydration.

Typically the dehydration of the carbinolamine is the rate-determining step of Schiff

base formation and that is why the reaction is catalyzed by acids. Yet the acid
concentration cannot be too high because amines are basic compounds. If the amine is
protonated and becomes non-neucleophilic, equilibrium is pulled to the left and
carbinolamine formation cannot occur. Therefore, many Schiff bases synthesis are
best carried out at mildly acidic pH.
The dehydration of carbinolamines is also catalyzed by base. This reaction is
somewhat analogous to the E2 elimination of alkyl halides except that it is not a
concerted reaction. It proceeds in two steps through an anionic intermediate.

The Schiff base formation is really a sequence of two types of reactions, i.e.
addition followed by elimination7.

1.2. Chemistry and Biological Importance of Schiff bases.

Schiff bases have a large number of synthetic uses in organic chemistry.
Acylation of Schiff bases8,9 by acid anhydrides, acid chlorides and acyl cyanides is
initiated by attack at the nitrogen atom and leads to net addition of the acylating agent
to the carbon-nitrogen double bond. Reactions of this type have been put to good use
in natural product synthesis.
Schiff bases appear to be an important intermediate in a number of enzymatic
reactions involving interaction of an enzyme with an amino or a carbonyl group of the
substrate. One of the most important types of catalytic mechanism is the biochemical
process which involves the condensation of a primary amine in an enzyme usually
that of a lysine residue, with a carbonyl group of the substrate to form an imine, or
Schiff base. Stereochemical investigation carried out with the aid of molecular model
showed that Schiff base formed between methylglyoxal and the amino group of the
lysine side chains of proteins can bent back in such a way towards the N atom of
peptide groups that a charge transfer can occur between these groups and oxygen
atoms of the Schiff bases. In this respect pyridoxal Schiff bases derived from
pyridoxal and amino acids have been prepared and studied from the biological point
3

of view. Transition metal complexes of such ligands are important enzyme models.
The rapid development of these ligands resulted in an enhance research activity in the
field of coordination chemistry leading to very interesting conclusions.
The carbon-nitrogen double bond of Schiff bases like the carbon-oxygen
double bond is readily reduced by complex metal hydrides8,9. Reduction of this type is
probably the most efficient and convenient method for the conversion of C=N into
amino compounds. Thus lithium aluminium hydride in THF at room temperature (or
in difficult cases at elevated temperature) smoothly reduces Schiff bases in high yield
(> 90 %) to secondary amines. Sodium borohydride is an equally effective reducing
agent and is preferred to lithium aluminium hydride because of its inertness to a wider
range of solvent media and because of its greater specificity in that other substituents
such as nitro or chloro reducible by lithium aluminium hydride are unaffected by
sodium borohydride. An even more effective reagent of this type is sodium
cyanoborohydride (NaBH3CN).
Also the base catalyzed condensation of acetyl chlorides (bearing an electron
withdrawing group and at least one hydrogen atom at the -position) with Narylaldimines occurs by initial acylation at the nitrogen atom and leads to -lactams
of interest in penicillin chemistry [Scheme 1; (1) + (2) (4)]10.

Iminium salt11 (R2C=N+R2) at the other extremes are very rapidly hydrolyzed
by water and have to be prepared under rigorously anhydrous conditions. The facility
of iminium salt hydrolysis has been put to use in a synthesis of secondary amines
from primary amines which involves conversion into the aldimine (R1CH=NR2) and
then by alkylation in to the iminium salt [R1CH=N+R2 (R3) X-] followed by hydrolysis
4

to give the secondary amines (R2NHR3). Because of the involvement of Schiff base
hydrolysis in a number of enzyme mediated processes, the detailed mechanism of
hydrolytic cleavage of carbon-nitrogen double bonds has been the subject of close
scrutiny both under in vivo and under in vitro conditions12. Imines hydrolysis is also a
key step in the Sommelet13, Stephen14, Sonn-MulIer15 and Gattermann16 aldehyde
synthesis.
Alkoxides add in the expected fashion to Schiff bases, giving the
corresponding -alkoxy amino compounds9. Addition of this type provides the key
step in an elegant 'one pot' stereo specific synthesis of penicillin intermediates which
can be further elaborated to new cephalosporin derivatives [Scheme 2; (5)(8)]17.
This involves the N-chlorination-dehydrochlorination of readily accessible penicillin
amides (5) using t-butyl hypochloride in methanolic borate at 0C to give acylimines
(7), which adds methanol from the less hindered -face to give the adduct (8) with
the desired stereochemistry.

Hydrogen sulphide reacts readily with N-substituted ketimines at low

temperature (-40 to 0C) to give initial adducts which rapidly react further to afford
gemdithiols. N-arylaldimines readily add thiols in the form of their metal salts to give
the expected 1,2-adducts which tend to disproportionate with net reduction of the
imines to the corresponding amine and concomitant formation of the disulphide8,9.
Reactions of Schiff bases with primary amines results in adducts which tend to
decompose to a new imine and primary amine8,18, the overall process corresponding
to imine exchange. The rate of imine exchange increases with increase in the basicity
of the primary amine effecting displacement. Sodamide18 reacts with aldimines with
5

formal replacement of the imidyl hydrogen to give amidines [Scheme 3; (9)(10)].

Iminium salts also readily add primary, secondary and tertiary amines with the
formation of aminals (gem-diamino compounds) or their quaternary salts19.

Schiff bases react in general with ethereal solutions of chloramines in a few hours at
room temperature to give moderate to high yields (40-70%) of diaziridines [Scheme
4; (11) (13)]20. This formal cycloaddition reaction has wide scope and is
applicable, using chloramine itself (or the comparable reagent hydroxalamine-osulphonic acid), as well as N-substituted chloramines to a variety of Schiff bases
derived from aldehydes and acyclic and cyclic ketones. In many cases the Schiff base
can be converted in situ into the diaziridine (yield 50-80%) by reacting the
corresponding carbonyl compound with ammonia or primary or secondary amines in
the presence of hydroxalamine-o-sulphonic acid or an N-substituted hydroxalamine-osulphonic acid. Diazidine formation is believed to result from initial nucleophilic
addition to the carbon-nitrogen double bond followed by eliminative ring closure
(11)(12)(13).

The addition of hydrogen cyanide to Schiff bases occurs readily and provides a
viable route to -amino nitriles, which can in turn be used as precursors for the
synthesis (via hydrolysis) of amino acids (Strecker synthesis)21.This reaction (which
is akin to cyanohydrin formation from carbonyl compounds) is usually carried out
6

using anhydrous hydrogen cyanide in inert solvents such as ether or benzene.

However, a more convenient procedure, which gives improved yields, is to use
sodium cyanide in a phosphate buffer. Trimethyl silyl cyanide has been
recommended22 as a safer alternative to hydrogen cyanide in the Strecker synthesis.
This reagent adds to Schiff bases in the presence of catalysts such as aluminum
chloride, zinc Iodide or trisacetylacetonate aluminum, to give -cyano-N-trimethyl silylamines which are not isolated but are converted by neutral hydrolysis into amino nitriles in high yield (80-90%). Alternatively, the cyanotrimethylsilyl adducts
can be converted by acidic hydrolysis directly into -amino acids.
Schiff bases lacking hydrogen atoms to the carbon-nitrogen double bond
react with Grignard and organolithium reagents (alkyl and aryl) analogously to
carbonyl compounds to give adducts which on hydrolytic work-up afford secondary
amines in good to excellent yield (60-90%) [Scheme 5]. Reaction occurs best with
aryl aldimines and provides the general method for the synthesis of secondary amines.
One disadvantage in this respect is that addition of organomettalic reagent is subject
to steric hindrance by Schiff base substituents (e.g. N-t-butyl) although sustituents on
the organometallic reagent do not appear to exert stearic effect. A further
disadvantage is that in some cases (particularly with Grignards reagent) the yields of
secondary amines are lowered by competing reductive dimerization of the Schiff base
(14)(16). The scope of this amine synthesis has been widened23 by the use of
alkylidine arenesuphenamides (14; R3=S Ar) as the Schiff base component. Reaction
of these substrates with organolithium reagents followed by hydrolytic work-up
affords moderate to good yields (40-80%) of secondary and tertiary alkyl primary
amines (16; R3=H).

Schiff bases incapable of enolization (i.e. lacking -hydrogen atoms) also

react with Reformatsky reagent with stereo specific addition to the carbon-nitrogen
double bond to give erythro--amino esters which can be isolated at low temperature
(-10C) but otherwise spontaneously cyclize, providing a useful synthetic route to lactams24. Alkali metals react with fully aryl-substituted Schiff bases to give N,C-bismetallated derivatives which can be sequentially alkylated or acylated at carbon then
at nitrogen to yield mixed C,N-alkylated, acylated, or alkylated-acylated products8,9,25.
The deprotonation of -C, N-bis-alkylated Schiff base can in principle give
rise either to l-aza-allyl anion or 2-aza-allyl anion. In practice deprotonation to the laza-allyl anion is preferred owing to its greater stability26.
However, 2-aza-allyl anions are readily accessible27 by the deprotonation of
Schiff bases containing N-alkyl substituents, and lacking -hydrogen atoms, at low
temperature (-70C) using lithium diisopropylamide in THF [Scheme 6; (18)(19)]
or by the conrotatory ring-opening of readily accessible N-lithioaziridines (20)(19).

In terms of their nucleophilic reactivity, anions of this type (21) have been
shown28 to react readily with aldehydes and ketones to afford alkylidene amino
alcohols which can be dehydrated (using thionyl chloride-pyridine) or hydrolyzed
(using aqueous hydrochloric acid) to provide synthetic routes to 2-azabuta-l,3-dienes
(23) and -amino alcohols (24) respectively [Scheme 7].

Reactions of anils with carbonyls-stabilized sulphonium ylides also provide a

useful method for the synthesis of -aminoalkenyl carbonyl compounds [Scheme 8;
(25)+(26)(29)]29. A course for these reactions involving initial nucleophilic
addition followed by ring closure and subsequent ring opening of an aziridine
intermediate is supported by the demonstration30 that, by analogy with the formation
of oxirans from carbonyl compounds, anils react with dimethoxyoxosulphonium
methylide to give moderate yields of aziridines.

[1+2] Cycloaddition reactions of carbenes and carbenoids to Schiff bases are

well documented31,32 and constitute the useful method for the synthesis of aziridines.
In its simplest form, cycloaddition of this type is exemplified by the reaction of
benzylidene -aniline with dichlorocarbene to give 2,2-dichloro-1,3-diphenylaziridine
9

in good yield (55%) [Scheme 9; (30)(31)]. Aziridines are also the products of the
reactions of Schiff bases with the Simmons-Smith reagent (methylene di-iodide/zinccopper couple)33.

Cycloaddition of Schiff bases to ketenes is highly stereoselective34, implying a

concerted process. However, a two-step mechanism involving a dipolar intermediate
[Scheme 10], adequately accounts for the observed stereoselectivity and is strongly
supported by mechanistic studies34,35 of -lactam (35) formation via intermediate of
the type (34)from Schiff bases (32) and ketens (33).

Cycloaddition of Schiff bases to simple isocyanates31,36,37 is preceded by the

reversible formation of a zwitter ion intermediate: [Scheme 11; (36)+(37)=(38)],
which in some cases36 can be isolated. The subsequent course of the reaction is then
dictated both by the stability of the zwitter-ion intermediate (38) (and hence
principally by the steric and electronic effects of the Schiff base substituents) and by
10

the operation of kinetic or thermodynamic control. Broadly, electron donating parasubstituents in the benzylidine moiety of the Schiff base (36) and low reaction
temperatures, with ensuing kinetic control, favors direct ring-closure of the zwitterion
(38) to the [2+2] cycloadduct (39). Conversely electron-withdrawing parasubstituents in the benzylidene nucleus of the Schiff base (36) and relatively high
reaction temperatures, with consequent thermodynamic control, promote dipolar
[4+2] cyclo addition of the zwitterion (38) with the Schiff base (36) or the isocyanate
(37) (present in equilibrium) to give the 2:1 and 1:2 adducts (40) and (41)36,37. The
cycloaddition reactions of Schiff bases with acyl isocyanates38 and acyl
isothiocyanates39 broadly follow the same pattern.

Schiff bases capable of imine-enamine tautomerism react with isocyanates to

afford acyclic adducts whose formation is rationalized in terms of [2+2] cycloaddition
of the isocyante (44) to the enamine tautomer followed by ring-opening of the
unstable adduct formed [Scheme 12; (42) = (43) (45) (46)]36,37,40.

11

The formal [2+2] cycloaddition of the carbon-nitrogen double bond to a

carbon-phosphorus double bond followed by retrocycloaddition accounts41 for the
Wittig-like reaction of Schiff bases (48) with arylidene phosphoranes (47) to afford
alkenes (51) and phosphinamines (52) via intermediates (49) and (50) [Scheme 13].

Schiff bases react readily with diazoalkanes (e.g. diazomethane) in the

presence of catalytic amounts of methanol or water to afford 2 -1,2,3-triazolines
[Scheme 14; (54)(55)]42. Electron-withdrawing groups on the Schiff base promote
this cycloaddition while electron-donating groups hinder it. These substituents effects
in conjuction with the catalytic effect of methanol and water and the high
regiospecificity observed, namely exclusive l,2,3-triazoline formation as oppose to the
alternative 1 -l,2,4-triazoline formation (54)(53), are consistent with a concerted
1,3-dipolar cycloaddition involving a polar transition state in which there is maximum
12

stabilization by overlap of the highest occupied molecular orbital (HOMO) in the

dipole with the lowest unoccupied molecular orbital (LUMO) in the dipolarophile31,42.

Nitrile oxides add to carbon-nitrogen double bonds of Schiff bases much more
readily than to carbon-oxygen double bonds, providing a general high yield route to
2 -1,2,4-oxadiazolines [Scheme 15; (56)+(57)(58)]42. The Schiff base component
(64) in cycloaddition of this type can be an acyclic N-alkyl or N-ary1 aldimine or
ketimine [(56; X = alkyl or aryl)] or can be integral with a ring as in the
cycloaddition of nitrile oxides with 2-pyrazolines to give fused structures [(57)+(59;
X=NR) (60; X=NR)].

The importance of electron-withdrawl in diene component is strikingly

demonstrated by the enhanced reactivity of hetero1,3-dienes in [4+2] cycloaddition in
comparison with their homo l,3-diene counterparts. Thus acyl and thioacyl
isocyanates and acyl isothiocyanates readily undergo l,4-cycloaddition with simple
Schiff bases to afford excellent yields of

1,3,5-oxadiazine and I,3,5-thiadiazine

derivatives [Scheme 16; (61) + (62) (63)]31,38,39,43.

13

The uncontrolled oxidation9 of a Schiff base with a peroxy acid results in

cleavage of the carbon-nitrogen double bond to give a carbonyl compound and a
nitroso compound, respectively. On the other hand, oxidation using peroxy acids at
low temperature (00C) affords an excellent synthetic route to oxaziridines [Scheme
17; (64 66)8,20,44.

Peroxy acid oxidation of Schiff base (69) [from primary amine (67) and
heterocyclic aldehyde (68) to an oxaziridine (70) followed by base-catalyzed
rearrangement has been shown45 to provide a model [Scheme 18] for the pyridoxal
pyrophosphate mediated enzymatic oxidative deamination of -amino acids to
pyruvic acids, which finds analogy in the well-known double bond transposition of
allylic alcohols via oxiran intermediates.

14

Initial hydroperoxide formation also accounts for the oxidative cleavage of

the Schiff base (71) from 2-aminopyridine and isobutyraldehyde, by potassium tbutoxide in dimethyl sulfoxide which affords acetone and 2-formamidopyridine (73)
in high yield (80-90%). The emission of blue light associated with this oxidative
transformation is consistent with the mechanism [Scheme 19] involving the formation
and cleavage of a dioxetan intermediate (72) and supports a similar mechanism for
bacterial luminescence46.

The oxidation of Schiff bases by metal based oxidants has been most
extensively investigated in the case of leadtetraacetate (LTA)44,47. LTA oxidation of
simple anils48 results in the formation of aldehyde, the arylamine, and the

15

corresponding azobenzene derivative. Formation of the amine and the azobenzene

derivative has been attributed to the involvement of a nitrene intermediate produced
by the ionic breakdown of an initially formed lead derivative [Scheme 20;
(74)(75)(76)+(77)(78)+(79)]. Where the anil contains, a suitable ortho
substituent (hydroxyl or amino) in the N-aryl ring. LTA oxidation leads to heterocycle
(benzimidazole, benzoxazole) formation [(74; Z = NHR or OH) (80; X = NR or
0)]44,47. Lead tetraacetate dehydrogenates N-unsubstituted aldimines to the
corresponding nitriles. Since the requisite aldimine can be generated in situ by
reaction of an aldehyde with ammonia, this constitutes a useful method for the
synthesis of nitriles from aldehydes.

The behavior of the carbon-nitrogen double bond of Schiff bases towards

reducing agents broadly parallels that of the carbon-oxygen double bond. Metalproton donor reagents (e.g. sodium, sodium amalgam, magnesium or aluminium in
ethanol; zinc or aluminium in aqueous alkali; zinc in acetic acid) smoothly reduce
Schiff bases (81), (prepared in situ from an aldehyde or ketone and an amine) to the
corresponding amines (82)8,9. Alkali metals in inert solvents such as ether or toluene
tend to promote reductive dimerization by a radical-coupling mechanism to afford
diamino compounds (83) as the major products8,9,49.

16

Schiff bases are readily reduced to amines by hydrogenation over platinum,

nickel or chromium catalysts8,9,50. Thus, anils are reduced to secondary amines in
essentially quantitative yield by hydrogenation over a platinum catalyst at 60oC. The
hydrogenation of mixtures of ammonia and primary amines with alkyl and aryl
aldehydes and ketones over nickel, platinum and palladium catalysts involves the in
situ reduction of Schiff bases and is a simple yet important general method for the
synthesis under mild conditions of primary and secondary amines in good to excellent
yield. This useful amine synthesis is applicable to polyfunctional molecules i.e.
synthesis of -amino acids (e.g. alanine) from -keto acids (e.g. pyruvic acid).
Asymmetric induction has also been demonstrated51 in the catalytic hydrogenation of
Schiff bases and leads to chiral amines in good optical yield. A recent new method for
the efficient conversion of Schiff bases into amines in excellent yield (80-90%) under
mild conditions involves reduction with alkyl silanes in the presence of transition
metal catalysts [e.g. PdC12, (Ph3P)3RhCI]52.
The carbon-nitrogen double bond is reduced electrolytically more readily
than the carbon-oxygen double bond. Schiff bases derived from aldehydes and
ketones are reducible electrolytically over a wide pH range to give the corresponding
17

amines. Thus the controlled potential reduction of a solution of cyclohexanone in

methylamine gives N-methyl cyclohexylamine in high yield. The electrolytic
reduction of N-substituted aryl aldimines and ketimines (84) occurs by the stepwise
addition of two electrons and gives initially a radical anion (85) and then by
protonation a redical intermediate (86) which can be further reduced to a secondary
amine (87) or dimerize to a 1,2-diamino compound (88), [Scheme 22]53-55.

Synthesis of Schiff bases have also been reported by different authors56-73.

Ugras et al74 have reported the synthesis, complexation, antifungal and antibacterial
activity studies of a macro cyclic Schiff base (Scheme 23).

(Scheme 23)
Preparation, physical characterization and antibacterial activity of Ni (II) Sciff
base complex (Scheme 24) was reported by Morad et al75.

18

(Scheme 24)
Elzahany et al76 have synthesized some transition metal complexes with Schiff
bases derived from 2-formylindole, salicylaldehyde and N-amino Rhodamine. The
Schiff base ligands were characterized by elemental analysis, IR, Mass, 1H NMR and
electronic spectra. The free ligands and their metal complexes were also screened for
antimicrobial activities against Bacillus cerens, Escherichia coli, Pseudomonas
aeruginosa, Staphylococcus aureus and Candida albicans. The results indicated that
the ligands do not have any activity, where as their complexes showed more activity
against the same organisms under identical experimental conditions

Synthesis and pharmacological studies of novel schiff bases of 4Hydroxy-6- carboxyhydrazino benzofuran was reported by Gopal Krishna et al77
(Scheme 25).

(Scheme 25)
19

4-Chloro-2-oxo-2H-chromene-3-carbaldehyde was made to react with

different anilines in rectified spirit to yield a series of Schiff bases (Scheme 26) of the
type 4-chloro -3- (substituted-phenylimino) methyl) -2H-chromen-2-one78. These
following compounds were characterized on the basis of their spectral (IR, 1H NMR)
data and evaluated for antimicrobial activity in vitro against fungi, gram positive and
gram negative bacteria.

(Scheme 26)
Yi YI and coworkers79 have reported the synthesis and color-tunable
fluorescence properties of Schiff base Zinc complexes which are used as
electroluminescent materials. These authors have reported that the Schiff base
(Scheme 27) Zinc complexes synthesized by them have good thermo stability,
solubility and film forming capability and can be used as organic electroluminescent
materials. These new complexes may afford the feasibility to realize full-color display
with materials based on similar molecular structures.

20

Vijey Aanandhi et al80 have reported the synthesis of a series of 1-(5substituted-2-oxoindolin-3-ylidene)-4-(substituted-pyridin-2-yl)

thiosemicarba-zide

derivatives (Scheme 28). These compounds were screened for in vitro antibacterial
and antifungal activity against B.subtilis, S. aureus, E.coli, P. aeruginosa, C. albicans,
and A. niger. All the compounds were reported to exhibit moderate to good
antibacterial and antifungal activity.

(Scheme 28)
Karaoglan et al81 have reported the synthesis and characterization of a new
Schiff base and its metal complexes. The Schiff base ligand were characterized by
FT-IR, 1H-NMR, UV-Visible, Mass spectra, elemental analysis and fluorescence
spectrophotometry.
Farias and Bastos82 have studied the electro chemical behavior of copper (II)
complexes of the schiffs base (111) N, N-ethylene bis(salicylidimine)in aqueous
phosphate (pH 7) by polarographic and voltametric techniques at a mercury electrode.
It is a symmetrical molecule and exhibits chiral properties.

(111)

Khalil et al83 were the first group to announce the possibility of using a Schiff
base as an acid-base indicator. This surprising phenomenon can be considered as an
interest due to the fact that Schiff bases are usually unstable in solutions and definitely
21

undergo hydrolysis. It was found that such a specific observation depends merely
upon the chemical structure and type of the substitute of amine that reacts with
aldehyde to give the Schiff base. The latter reagent 4{(4-dimethylamino-benzylidine)amino}-benzene sulfonamide was synthesized from the condensation of sulfanilamide
with p-dimethylaminobenzaldehyde. The reagent solution shows a reproducible
change in its color due to the addition of acid and base. A UV-Visible spectroscopic
characterization and acid-base equilibrium study of the reagent for its possible use as
an indicator were investigated. The results show that the reagent is amphoteric which
possesses four ionization constants Kal, Ka2, Kb1 and Kb2 of weak dibasic and
diacidic properties. It was concluded that the benzyl sulfonamide group plays a key
role in the stability of the reagent towards hydrolysis and also for indicator
characteristics through breaking the conjugation. Jamil and coworkers84 have reported
the synthesis, characterization and antimicrobial activities of novel organotin schiff
base compounds.
Metal complexes of Schiff bases derived from 2-furancarboxaldehyde and ophenylenediamine and 2-thiopheneacarboxaldehyde and 2-aminothiophenol was
reported by Geindy et al85.These authors have reported the ligand dissociation as well
as the metal-ligand stability constants for these complexes. The synthesized ligands,
in comparison to their metal complexes were also screened for their antibacterial
activity against bacterial species, Escherichia coli, Pseudomonas aeruginosa and
Staphylococcus Pyogones as well as fungi (Candida). The activity data reveal that the
metal complexes are found to be more potent antibacterial than the parent Schiff base
ligand against one or more bacterial species.

22

A series of biologically active pyrazine-derived Schiff base ligands (115, 116)

have been synthesized by the condensation reaction of 2-aminopyrazine with
salicylaldehyde and acetamidobenzylaldehyde. Then their Co(II), Ni(II) & Zn (II)
complexes have been prepared. The biological evaluation of the simple uncomplexed
ligand in comparison to their complexes have been determined against bacterial
strains namely Escherichia coli, Staphylococcus aureous and Pseudomonas
aeruginosa86.

SARI and coworkers87 have reported the synthesis and antibacterial activities
of some new amino acid-Schiff bases as follows:

(117)

(118)

(119)

Mixed ligand transition metal complexes of Cu+2, Ni+2 and Co+2 ions with
Schiff base ligands (Scheme 29) derived from the condensation of o-hydroxy
benzaldehyde with amino phenols and nitrogen donor amine bases was reported by
Saidul Islam et al88 The authors have also studied the antibacterial and antifungal
activities of the compounds.
23

(Scheme 29)
Daniel Thangadurai and Son-Ki Ihm89 have reported the synthesis,
characterization, catalytic and antibacterial studies of chiral Schiff base Ruthenium
complexes. These authors have tentatively proposed an octahedral structure for all the
new complexes. The catalytic and antibacterial activities of these compounds have
also been reported (126).

(126)
90

Baluja et al

have studied the biological activities of the following Schiff

base and metal complexes (Scheme 30).

(Scheme 30)
24

Metal complexes ML2Cl2 where M is Fe (II), Co (II), Ni(II), Cu(II), Zn(II) or

Cd(II) and Li is the Schiff base formed by the condensation of 2thiophenecarboxaldehyde

with

2-aminopyridine,

N-(2-thienylmethylidene)-2-

aminopyridine (TNAPY) have been reported by Spinu et al91 (130).

(130)

Thilagavathi and coworkers92 have reported the synthesis (Scheme 31) of 3{4[4-(benzylideneamino) benzenesulfonyl]-phenyl}-2-phenylquinazolin-4(3H)-one.

(Scheme 31)
Synthesis, characterization and electrochemical behaviour of Cu, Co, Ni and
Zn complexes derived from acetylacetone and p-anisidine was reported by Raman and
coworkers93. These authors have observed that the complexes synthesized by them
show fairly good antimicrobial activity (133).

(133)

New Schiff base of the type, 2-[4-methyl-2-oxo-2H-chromen-7-yl)oxy]-N1(substituted methylene)acetohydrazides were synthesized94 by the condensation of
aryl/hetero aromatic aldehydes with 2-[(4-methyl-2-oxo-2H-chromen-7-yl)oxy]
acetohydrazides under conventional and microwave conditions and characterized

25

through IR, 1H NMR and mass spectral data. The synthesized compounds have been
screened for antimicrobial activity.
Raman et al95 have reported the synthesis of the following Schiff base ligand
(134) These authors have also studied the DNA cleavage and antimicrobial activity of
the Schiff base transition metal complexes .

(134)

Chittilappilly and Yusuff

96

have reported the synthesis, characterization and

biological properties of ruthenium Schiff base complexes derived from 3-hydroxy

quinoxaline-2-carboxaldehyde and salicylaldehyde (135).

(135)

Two new Schiff base ligand containing cyclobutane and thiazole rings 4-(1methyl-1-mesitylcyclobutane-3yl)-2-(2,4dihydroxybenzylidenehydrazino)Thiazole
and

4-(1-methyl-1-mesitylcyclobutane-3-yl)-2-(2-hydroxy-3-methoxybenzylid

enehydrazino)thiazole and their mononuclear complexes with Co(II), Cu(II),Ni(II)

and Zn(II) in EtOH was reported by Cukurovali and co workers97 (Scheme 32).

26

(Scheme 32)

Hearn and Cynamon98 have reported the synthesis and antitubercular activity
of Schiff base of the following type.

(142)

Nair and coworkers

99

have studied the synthesis and antibacterial activity of

some Schiff base complexes. The Schiff bases (143) showed greater activity than their
metal complexes .

(143)

27

Vanden Ancker et al100 have reported the synthesis of the following bis-imine
Schiff bases (Scheme 33). These authors have claimed that bis-imine Schiff bases are
obtained in high yield (>95%) when aliphatic diamine/aldehyde condensation
reactions are carried out under solvent-free conditions or in polypropylene glycol
(PPG) as a recyclable reaction medium with negligible waste.

(Scheme 33)

Potentially heptadentate tripodal Schiff-base ligand (146) have been

prepared101 and characterized by various spectroscopic methods such as IR, FAB-MS
and NMR.

Dong-Hoon Won and coworkers102 have studied the synthesis and crystal
structure of the following Schiff base macrocycles bearing thiophene (147).

28

(147)

Shabani et al

103

have reported the synthesis, characterization and anti-tumor

activity of Iron Schiff base complexes. Iran Sheikhshoaie and Samira104 have reported
the synthesis, characterization and nonlinear optical properties of the following Schiff
bases (Scheme 34).

(150)

(151)

29

(Scheme 34)
Gao and Zheng105 have reported the synthesis of optically active Schiff base
ligand (155) derived from condensation of 2-hydroxyacetophenone and 1,2
diaminocyclohexane.

(155)

Synthesis of Schiff bases of naptha[1,2-d]thiazol-2-amine and metal

complexes of 2-(21-hydroxy)benzylideneaminonapthathiazole as potential
antimicrobial agent (Scheme 35) was reported by Faizul and coworkers106.

30

(Scheme 35)
Ibrahim and Sharif107 have studied the synthesis, characterization of following
Schiff bases (Scheme 36) which can be used as fluorometric analytical reagents.

(Scheme 36)
31

Gudasi et al108 have reported the synthesis, characterization and biological

studies of dioxouranium and thorium complexes of Schiff base (Scheme 37) derived
from 2-amino pyridine and acetophenones.

More et al109 have reported the synthesis of the following Schiff base (169).
These authors have studied the proton ligand stability constant of the Schiff bases and
the formation constants of their transition metal complexes.

(169)

Bag et al

110

have synthesized a series of Schiff bases of benzidene and

examined the mercuration reaction (Scheme 38).

(Scheme 38)
32

Following two (174,175) new Schiff bases and transition metal complexes
derived from 2, 3-diminopyridine (DAPY) and ortho-vanillin have been
synthesized111 and characterized by elemental analysis, magnetic susceptibility
measurements, IR and NMR spectra. The Schiff bases and most of the metal
complexes display antibacterial activity.

(174)

(175)

Rajendran and Karvembu112 have reported the synthesis of following Schiff

bases derived from 3-amino-2Hpyrano[2,3-b]quinolin-2-ones. The synthesized Schiff
base (Scheme 39) compounds were screened against the fungal strains, such as
Aspergillus niger and Fusarium sp..

(Scheme 39)
Rosu et al113 have reported the synthesis of Cu complexes derived from
following Schiff base ligands (178, 179) obtained by the condensation of 2-hydroxybenzaldehyde or terephthalic aldehyde with 4-aminoantipyrine.

33

(178)

(179)

Jarrahpour et al114 have reported the synthesis of twelve new bis-Schiff bases
of istatin, benzylisatin and 5-fluoroisatin by the condensation with primary aromatic
amines (Scheme 40).

(Scheme 40)
34

The UV visible spectra of some Schiff bases derived from 2-aminopyridine

and 2-aminopyrazine have been investigated in acetonitrile and toluene. The
compounds (183, 184) were in tautomeric equilibrium in polar and non polar solvents.
This was suggested by Asiri et al115.

(183)

(184)

Sivakumar et al116 have reported the proton dissociation constant of the ligand
and the stability of the complexes of Co(II), Ni(II), Zn(II), Cd(II), Hg(II) and Pb(II)
ions with 2-phenyl-3-(2-hydroxy-5- benzylidine)-quinazoline-4-(3H)-one. The
proton-ligand and metal-ligand stability constants of the complexes have been
determined pH metrically by the Calvin Bjerrum titration technique (Scheme 41).

(Scheme 41)
Wei et al117 synthesized a pair of iso structural azido or thiocyanato bridged
centre of symmetric dinuclear copper (II) complexes derived from the Schiff base
ligand, 4-nitro-2-[(2-diethylaminoethylimino)methyl]phenol (188). These compounds
are characterized by elemental analysis, IR spectra and single X-ray diffraction. The
antimicrobial activities of the complexes have been tested.

(188)

Natarajan Raman et al118 have reported the synthesis of a novel 14-membered

35

macrocyclic Schiff base (189) derived from 3-cinnamalidene- acetanalide and ophenylenediamine which acts as a tetradentate and strongly conjugated ligand to form
a cationic solid complex with Cu(II)/Ni(II)/Co(II) and /Zn(II). The ligand and the
complexes were characterized by the usual spectral and analytical techniques. The
antimicrobial tests were also recorded and gave good results in the presence of metal
ions in the ligand system.

(189)

An investigation dealing with the impact of following Schiff base (190) derived from
anthranilic acid and acetoacetanilide and its copper complex on instar larvae of
Spodopetra litura was done by Raman et al119.

(190)

Twenty Schiff bases of 2-amino-5-1,3,4-oxadiazoles have been synthesized

with different aromatic aldehydes120.The antibacterial properties of the compounds
were investigated against Proteus mirabilis and Basilus subtilus. A series of 4substituted-emoni-methyltetrazolo[1,5-a]quinoline with appropriate aromatic amine
by refluxing in dioxane. They have been evaluated for their anti-inflammatory and
antimicrobial activities121 (Scheme 42).

36

(191)

(192)

(193)

(Scheme 42)
Biyala et al122 have studied the synthesis of mono basic bidentate Schiff base
complexes of palladium and platinum from 1H-indol-2,3-dione thiosemicarbazone.
These complexes were characterized on the basis of elemental analysis, molecular
weight determination, 1H NMR and UVspectral studies. Antimicrobial effects of both
the ligands (194,195), and their complexes on different species of pathogenic fungi
and bacteria have been recorded and these are found to possess significant fungicidal
and bactericidal properties.

(194)

(195)

Synthesis, crystal structures and antimicrobial activities of two thiocyanatobridged

dinuclear

diethylaminoethlimino)

copper

complexes

methyl]

phenol

derived
and

from

2,4-dibromo-6-[(2-

4-nitro-2-[(2-thylamino

ethyl-

imino)methyl]phenol was proposed by Zhe Hong123. These complexes have been

characterized by physico-chemical and spectroscopic methods. These are found to be
antimicrobial. In-vitro antibacterial and antifungal activities of five different amino
acid Schiff bases derived from the reaction of 2-hydroxy-1-napthaldehyde with
glycine, L-alanine, L-phenylalanine, L-histidine, Ltryptophaneandthe manganese(III)
complexes of these bases were investigated by Sakiyan et al124.The in-vitro activities
against some Gram positive and Gram negative bacteria and fungi were determined.
Tarafder et al125 have reported the synthesis of complexes of a tridentate
schiff base from the condensation of S-benzyldithiocarbazate with salicylaldehyde.
These complexes have been characterized by elemental analysis and spectral analysis.
Square planar structures are proposed for the Ni and Cu complexes. These authors
have also studied the antimicrobial tests which indicate that Schiff base and five of the
37

metal complexes of Cu, Ni, U, Zr and Sb are strongly active against bacteria. The
Schiff base exists in tautomeric form (196,197).

Ispir et al126 have reported the synthesis of Schiff base ligands (198, 199)
containing -SiOH3 or -SiOCH2CH3 groups, 4-{[(3-trimethyoxysilanepropyl) imino]
methyl}benzene-1,3-diol and 4-{[(3-triethoxysilanepropyl) imino] methyl}benzene1,3-diol from 2,4-dihydroxy-benzaldehyde and 3-amino propyltrimethoxysilane and
3-aminopropyl-triethoxysilane.

(198)

(199)

Singh127 proposed a reaction of bis(triorganotin)oxide with Schiff bases

derived by condensation of heterocyclic ketones, 2-acetylfuran and 2-acetylthiophene
with various sulfa drugs. The structures of the complexes have been established by
spectral studies. Molar conductance measurements were carried out for the Schiff
bases. The results of antimicrobial effects of some representative complexes on
different species of pathogenic fungi and bacteria have also been recorded.
Raman et al128 have reported the synthesis of Schiff bases (200) of 4aminoantipyrine neutral complexes of Cu (II) from salicylidine-4-aminoantipyrine

38

and PhNH2 substituted anilines. These authors confirmed their structure using IR,
UV- visible, 1HNMR and 13C- NMR spectra.

(200)

Schiff bases of some biological interest have also been studied by some other
authors129-134.

39

AZETIDINONES (BETA-LACTAMS)
The -lactams are 4-membered cyclic amides derived from 3-aminopropanoic
acids. Though the first member synthesized by Staudinger129 in 1907, the -lactams as
a class acquired importance since the discovery of penicillin which contains -lactam
unit as an essential structural feature of its molecule, this interest continued unabated
because of the therapeutic importance of -lactam antibiotics and recent finding of
new naturally occurring -lactams. As a result of vigorous research, a vast literature
has been accumulated over the years, and the chemistry of azetidinones continues to
be blossoming field.
Recent years have seen a resurgence of interest in the development of stereo
and enatioselective methodologies. The utility of azetidinones as synthons for various
biologically active compounds, as well as their recognition as cholesterol absorption
inhibitors and enzyme inhibitors has given impetus to these studies.
The -lactams are 4-membered cyclic amides derived from 3-aminopropanoic
acids. Though the first member synthesized by Staudinger129 in 1907, the -lactams as
a class acquired importance since the discovery of penicillin which contains -lactam
unit as an essential structural feature of its molecule. In the late 1990s, several groups
reported novel methodologies for the synthesis of azetidinones of potential biological
activities by applying known methods130-147.

2.1. Chemistry of Azetidinones

In the literature, monocyclic -lactams are usually referred to as azetidin-2ones or 2-oxoazetidine, based on the nomenclature of the parent heterocycle,
azetidine. However, the trivial names penam148 for the fused -lactam (201 a) and
cepham for the bicyclic system (202 a) are also used149,150. Similarly, the term openam, o-cepham, azapenam and azacepham were coined for the bicyclic -lactam
(201 b), (202 b), (201 c) and (202 c) respectively. This trivial system of nomenclature
is inadequate, especially in the case of fused -lactams having no bridge head nitrogen
atom, and in those having no heteroatom at position 1 or alterations in the positions of
the hetero atom of the non -lactam ring. This discrepancy can be removed by
adopting a new system in which fused -lactams (203) and (204) may be called
Alkanam and isoalkanam respectively. Thus, -lactams containing 7, 8 and 9
40

atoms in the bicyclic system (203) may be given generic names, heptanam, octanam,
nonanam and so on, using the corresponding latin roots. The numbering system as
shown in (201 d) and (202 d) is in conformity with the convention followed in the
case of penam-cepham nomenclature. Thus, the conventional penam will be termed as
1-thiaheptanam, and cepham as 1-thiaoctanam according to this system. Similarly, the
fused -lactams of the type (204) may be termed as isoheptanam, isoctanam,
isononanam and so on, depending on the number of atoms in the bicyclic system. The
numbering of ring atoms in this case may be the one used for azetidin-2-ones, and is
shown in (205).

(a) Z = S, (b) Z = O, (c) Z = NH, (d) Z = CH2

A bicyclic -lactam containing a double bond in the ring system may be given
the corresponding generic name derived from the collective name Alkenam or
Isoalkenam depending on the mode of fusion of the rings. For stereo description of
the molecule, the terms and denoting the configuration of the substituents,
which may be below or above the plane of the -lactam ring, may be used as in case
of steroids.

Construction of -lactam ring

There are diverse synthetic routes to -lactams151-155 and in principle the 4-membered
heterocycle could be constructed by the formation of one, two, three or all four bonds
of the ring system during the process of cyclisation156-171.
41

Cyclisation of 3-aminopropanoic acid derivatives

Five 1,3,4-triarylazetidin-2-ones (207) were prepared by treating (206) with
benzenesulfonyl chloride and alkali. Saponification of 2, 2-disubstituted-3-benzamido
propanoic acid esters (208) was found to give -lactams besides the acid
derivatives172. The cyclisation is possibly initiated by removal of the amidic proton,
followed by Dieckmann reaction (Scheme 43).

(Scheme 43)
Bicyclic -lactams such as penicillins173,174, cephalosporin175 analogs176,177 and
the compound (210)178 were synthesized by this method, using carbodimides as
Cyclising agents.

Blinkovsky et al179 have suggested the synthesis of beta-lactam antibiotics

containing alpha-aminophenylacetyl group in the acyl moiety catalyzed by Dphenylglycyl-beta-lactamide amidohydrolase.
42

Carlos Cativiela et al180 have reported the asymmetric Synthesis of Betalactams by Diastereoselective Alkylation of Chiral 2-Cyano Esters.

Addition of imines
The first -lactam was prepared by the ketene-imine interaction. Usually
ketenes are generated in situ by dehydrohalogenation of suitable acetyl chlorides in
the presence of a tertiary base. Also, photolysis and thermal decomposition of
diazoketones were employed for generating ketenes, which were trapped by imines to
give -lactams. Thermal fragmentation of acetylenic ethers to aldoketenes was also
reported.
The choice of ketene precursor is important, because it gives -lactams with a
suitable group at the carbon atom to the -lactam carbonyl function154. The
structural requirements of the imines are difficult to define due to the inconsistency in
the results obtained from different procedures. Imidylchlorides, o-alkyloximes and
phenylhydrazones did not give azetidin-2-ones. Addition of diphenyl ketenes on acyl
hydrazones is reported to give -lactams. Imines such as (210 a) and (210 b) gave
(211 a) and (211 b), on treatment with diphenyl- and dimethylketenes respectively153.
Tert-butylcyanoketenes with imino ethers gave -lactam (211 c)181. Diphenyl ketenes
with imines (210 d) gave -lactams (211 d), but their reactivity and yields varied
considerably with change in the substituents in the aromatic ring. Conjugated
diimines182,183and carbodiimides154 also gave -lactams with suitable ketenes (Scheme
44).

(a)

R1 = morpholine, R2 = H, R3 = Ph, R4 = R5 = Me or Ph

(b)

R1 = Ph, R2 = MeS-, R3 = Ph, R4 = R5 = Me or Ph

(c)

R1 = H, R2 = -OMe, R3 = R4= t-Bu, R5 = CN

(d)

R1 = R3 = Substitutedphenyl, R2 = H, R4 = R5 = Ph
(Scheme 44)

43

Reactions of isocyanates
Diazomethane was found to give -lactams (213) when treated with phenyland p-bromophenylisocyanates184. Indolyl-3-isocyanate reacted similarly152.

(Scheme 45)

Ring expansion of 3-membered rings

The aziridine (214) in the presence of thionylchloride or oxalylchloride
rearranges to -lactam (215) in benzene, possibly via a mixed anhydride which
undergoes ring expansion. The conversion is stereospecific and yields are good185
(Scheme 46).

(Scheme 46)

Ring contraction of 5-membered rings

Photolytic Wolf rearrangement of 3-diazopyrrolidine-2,4-diones (216), in the
presence of tert-butylcarbazate afforded -lactams (217)186,187. This method was
extended to the synthesis of azetidin-2-one (218), which was found to be biologically
inactive188. The fused system (219) under similar conditions produced (220), which
was found to be highly unstable189 (Scheme 47).

44

(Scheme 47)

Passerini reaction
The reaction of carbonyl compounds with 3-aminopropanoic acids, followed
by treatment with a suitable isocyanide afforded -lactam derivatives. This is an
extension of the Passerini reaction and it was useful for the preparation of monocyclic
and bicyclic -lactams (222) and (223) respectively. The reaction envisages formation
of a cyclic compound (221) which on transannular acyl migration gave the -lactam
(222). It is note worthy that the configuration of newly formed asymmetric center in
the penicillin analog (223) is predetermined by the steric disposition of the reacting
molecule (Scheme 48).

(Scheme 48)

45

Rearrangement reactions
There were several cycloadducts which undergo thermal or photochemical
fragmentation, generating ketenes and imines which recombine to give -lactams.
This method is of limited use because of the drastic conditions involved and possible
side reactions. Beckmann rearrangement of o-sulfonyloximes (224) was reported to
give novel -lactams (225) but now the revised structure has been proposed154.

Reactions of Beta-lactams
Cleavage of the -lactam bond
The -lactam bond undergoes rupture in the presence of an alkali, acid and lactamase, yielding 3-aminopropanoic acids. By selective degradation the natural
-lactams could afford useful amino acids. In the presence of dry hydrogen chloride, a
-amino acid hydrochloride is generated. For example, the compound (226) gave
(227) on treatment with hydrogen chloride in methylene chloride190. Similarly, the lactam may be cleaved by imines191 (Scheme 49).

(Scheme 49)

Cleavage of the 2,3-bond in azetidin-2-ones

1-Haloazetidin-2-ones (228) undergo photolytic or thermolytic cleavage to
give isocyanates (229) capable of undergoing secondary cyclisation under suitable
conditions192. Similarly, 3-azidoazetidin-2-one (230) on refluxing in diglyme,
underwent ring expansion through 2,3-bond cleavage193(Scheme 50).
46

(Scheme 50)

Cleavage of 5,6-bond in penicillin

Rearrangement of penicillin to penilloic acid (233) involves cleavage of the
5,6-bond186 (Scheme 51). Similar bond cleavage was observed in penicillin-1oxide195.

(Scheme 51)

Cleavage of the 1,4-bond in azetidin-2-ones and collapse of the bridge

in bicyclic Beta-lactams
-Lactams bearing a C-4 hetero atom are unstable and easily undergo 1,4-bond
cleavage196.

For

example,

the

4-mercaptoazetidin-2-one

(234)

changes

to

isothiazolinone (235) in 40% yield, on treatment with dimethylsulfoxide197 (Scheme

52).

(Scheme 52)
47

Fragmentation of Beta-lactams
Monocyclic Beta-lactams on photolysis or thermolysis break up into ketenes
and imines or alkenes and isocyanates, depending on the substituents present in the
molecule and which ever fragmentation is energetically profitable198. This process is
essentially a case of retrocycloaddition. Reagent induced fragmentation leads to
diverse products, depending on the substituents and reagents used. Fragmentation of
penicillin199 and cephalosporin200 occurred on treatment with trifluoroacetic acid, the
fragments being amido ketenes, and 2-thiazoline and 2-1,3-thiazine derivatives
respectively. Sometimes the fragment formed as primary products may undergo
secondary reactions. For example, -lactam (236) on retro Michael reaction, gave
(237) and subsequently (238) and (239)201 (Scheme 53).

(Scheme 53)
Enzyme catalyzed fragmentation of benzylpenicilin was reported202,203. It is
noteworthy that the azido group in -lactam (240 a) on reduction with Adams
catalyst and subsequent-acylation with phenoxyacetylchloride and triethylamine
afforded the 6-phenoxy compound (240 c)204. Such an unusual result may be
explained only on the assumption that the 6-amino compound (240 b) undergoes
fragmentation and generates a 2-thiazoline, which then reacts with phenoxyacetyl
chloride and triethylamine in the usual way.

48

2.2. Biological Importance of Beta-lactam Derivatives.

2.2.1. Beta-lactam derivatives as antimicrobial agents.
Azetidine and their derivatives have been extensively explored for their
applications in the field of medicine205-209. Likewise, azetidin-2-ones are of great
importance because of -lactam derivatives as an antibacterial agent210-214. Recently,
incorporation of these compounds have witnessed a great upsurge in the treatment of
tuberculosis and other chemotherapeutic diseases215. Sharma et al216 reported
synthesis and antibacterial activity of some N-sulphonamoylphenylamino-3-chloro-4phenylazetidin-2-ones. Most of the compounds exhibited significant antibacterial
activity.Comp.1 [4(5,6 dimethoxy pyrimidino sulphonamoyl)phenylamino]-3-chloro4-phenylazetidin-2-one (241) has been found to be very potent compound against E.
coli.

A series of 1-[5-(N10-phenothiazinomethyl)-1,3,4-thiadiazol-2-yl]-4-substituted-2azetidinones as antifungal agents have been reported by Rawat et al217. All the
compounds were screened for their antifungal activity against the fungi Candida
albicans, Rhizopus oryzae and Crysosporium pannical218. The fungicidal data
indicated that all the compounds were moderately to highly toxic. The toxicity of
compounds depends upon the nature and position of the substituents at the aryl
moiety. Compound (242) displayed promising antifungal activity.

Shah et al219 synthesized azetidinones (243) from hydrazine thieno [3,2d]pyrimidines as potential antimicrobial agents. All the products have been evaluated
for their in vitro growth inhibitory activity against several microbes like B. megatilis,
49

B. subtilis, E. coli, A. aerogens and A. awamori. Most of the compounds exhibited

maximum activity in the range of 21-27 mm against A. aerogens. Other compounds
showed either moderate or less activity against these organisms. None of the
compounds synthesized was found to exhibit significant activity against B. subtilis.

Parmar et al220 reported synthesis of azetidinones from hydrazinopyrimidine as

potential antimicrobial agents. All the products were evaluated for their in vitro
growth inhibitory activity against several microbes like B. megaterium, B. subtilis, E.
coli, P. fluorescens and A. awamori. All the compounds exhibited mild to moderate
antimicrobial activity against all microorganisms except (244) which exhibited
promising activity with ampicillin and chloramphenicol against P. fluorescens.

Antimicrobial activity of azetidin-2-ones has also been reported by various

221-228

authors

50

Mechanism of action of Beta-lactam derivatives as Antimicrobial

agents.
Beta-lactams inhibit cell wall synthesis (Figure No. 1). The peptidoglycan is
composed of sugars and amino acids. The sugar components consist of alternating
residues of N-acetyl glucosamine and N-acetyl muramic acid residues. Peptide chain
of 3-5 amino acids is attached with N-acetyl muramic acid. The peptide chain can be
cross linked to peptide chain of another strand forming mesh like layer by
transpeptidase. Beta-Lactams bind to PBPs which catalyse transpeptidation reaction.
They inhibit transpeptidation (final stage in the synthesis of cell wall) (Figure No. 2).

Figure No. 1

51

(Figure No. 2) Peptidoglycan synthesis

2.2.2. Beta-lactam derivatives as antitubercular agents.

Synthesis and antitubercular activity of Beta-lactam derivatives229-232, has been
reported by different authors. The representative compounds were tested in vitro for
their anti-tubercular activity against M. tuberculosis H37Rv. The data were compared
with standard drug Rifampin. All the compounds showed moderate antitubercular
activity against M. tuberculosis.
Patel et al233 have reported synthesis and antitubercular activity of 2-[4-(4substitutedphenyl)-3-chloro-2-azetidinon-1-yl]-4-[2-(4-chlorobenzene

sulphona

mido)-phenyl] thiazoles (245). Primary screening of the compounds for antitubercular

activity was conducted at 12.5 mcg/ml against M. tuberculosis H37Rv. Compounds
demonstrating at least 99% inhibition in the primary screening were tested at lower
concentrations against this microorganism to determine actual minimum inhibitory
concentration. The antitubercular activity data showed that most of the azetidinone
derivatives exhibited 100% inhibition in the primary screen at 12.5 mcg/ml
concentration.
52

Vashi et al234 have reported synthesis and antitubercular activity of 2azetidinones bearing thymol moiety. The products displayed moderate to good
tuberculostatic activity. Synthesis and antitubercular activity of 2-(4-aryl-3-chloro-2azetidinon-1-yl-amino)-6-(4-chlorophenyl)-5-cyano-3-N-methyl-3,4dihydropyrimidin
-4-ones is reported by Modha et al235. All the products displayed mild to moderate
antitubercular activity against M. tuberculosis. Compound (246) was the most active
member of this series.

2.2.3. Beta-lactam derivatives as anti-inflammatory agents.

Several such comp. like 1-[5-(carbazolylmethyl)-1,3,4-thiadiazol-2-yl)]-4(substituted phenyl) -3-chloro-2-oxo-azetidines have been synthesized and evaluated
for their anti-inflammatory activity by Srivastava et al236. All the compounds
displayed mild to moderate anti-inflammatory activity except compound (247) that
showed anti-inflammatory activity that was comparable to standard drug
phenylbutazone.

53

Several comp. like 1-[5-(N10-2-chlorophenothiazinomethyl)-1,3,4-thiadiazol2-yl]-4-(substituted phenyl)-3-chloro-2-oxoazetidines have been synthesized and

evaluated for their anti-inflammatory activity by Srivastava et al237. All the
compounds tested for anti-inflammatory activity exhibited mild to moderate activity.
The compound (248) was the most potent and active member of this series. It
displayed comparable anti-inflammatory activity but lesser than the standard
phenylbutazone.

2.2.4. Beta-lactam derivatives as anticancer agents.

Shah. et al238 thoroughly analysed the mechanism of inhibiton of human
leukocyte elastase (HLE) by a monocyclic lactam. This work led to the identification
of 4-[(4-carboxyphenyl)-oxy]-3,3-diethyl-1-[(phenylmethyl) amino] carbonyl] -2azetidinone as the first orally active inhibitor of human leucocyte elastase (HLE).
Analogues w i t h different substituents on the urea-N were synthesized and evaluated
for their activity in vitro against HLE as well as in vivo in a hamster lung
haemorrhage model. Compounds with a methyl or methoxy group in the para position
of the benzene ring were very potent in both assays. Park et al239 synthesized and
evaluated two known phenolic metabolites of paclitaxel. The C3-phenolic metabolite
of paclitaxel was prepared from 7-(triethylsilyl)-baccatin III and enantioenriched Nbenzoyl-2-azetidinone. The C2 phenolic metabolite was synthesized from paclitaxel
via selective C2 debenzoylation reacylation. Both the metabolites were found to have
good anticancer activity. Spletstoser et al240 synthesized and evaluated a novel
paclitaxel derivatives. The synthesis involved the preparation of an azide-containing
C-13 side chain through a Staudinger cycloaddition followed by a growth and a
variety of other cell lines. Compounds inhibited tubulin polymerization with potencies
commensurate with their cytotoxic activity and a more lipase-mediated kinetic
resolution through which azetidinone in 99% cc was obtained. Coupling of the
54

enantiopure side chain precursor to 7-TES-baccatin and subsequent silylether

deprotection afforded 3-(4-azidophenyl)-3-dephenyl paclitaxel, which was shown to
be as active as paclitaxel in tubulin assembly and cytotoxicity assays. A series of
novel 1,4-diaryl-2-azetidinones was synthesized by Sun et al241 using stereo specific
staudinger reaction as conformationally restricted analogues of combrestatin because
molecular modeling studies suggested close geometric similarities. They were
evaluated for cytotoxicity against a number of human tumor and normal cell lines.
Strong potencies were observed, with the best compound exhibiting IC (50s) of 2574 nm against human neuroblastoma IMR 32 cell soluble aniline-containing analogue
was found to be very effective in inhibiting the growth of AR42J rat pancreatic
tumors when transplanted into the nude mice. Boge et al242 synthesized and evaluated
novel cyclohexyl analogues of taxol and taxotere. Compound 2-(cyclohexyl
carbonyl)-2-debenzoylbaccatin was prepared from baccatin by hydrogenation.
Subsequent coupling of 2-(cyclohexyl carbonyl)-2-debenzoylbaccatin with N-t-BOC3-[(tert-butyldimethylsilyl)oxy]-4-phenyl-2-azetidinone, followed by removal of the
protecting groups afforded 2-(cyclohexyl carbonyl)-2-debenzoyl taxotere. In a similar
synthetic sequence, 3-cyclohexyl-3-dephenyl taxol was prepared from N-benzoyl-3[(tert-butyldimetylsilyl) oxy]-4-cyclohexyl-2-azetidinone and (triethylsilyl) baccatin.
The taxol analogue, in which all three taxol phenyl groups were substituted by a
cyclohexyl moiety, was synthesized in one step from taxol via hydrogenation. All
analogues exhibited strong activity in the microtubule assembly assay and
cytotoxicity comparable to taxol against B16 melanoma cells. Different authors243-251
have reported the Synthesis of anticancer Beta-lactams & their mechanism of action.

2.2.5. Beta-lactam derivatives as cholesterol lowering agents.

Fluorescent analogues of the cholesterol absorption inhibitor (CAI), have been
synthesized by Burnett et al252 as enantiomers. Biological testing revealed that they
were potent cholesterol absorption inhibitors (CAIs) and were suitable tools for the
investigation of the azetidinone cholesterol absorption inhibiting mechanism of
action. Ezetimibe253, (1-(4-fluorophenyl) -(3R) -[3-(4-fluorophenyl) -(3S) hydroxylpropyl] - (4S)-(4-hydroxyphenyl)-2-azetidinone) potentially and selectively inhibited
the intestinal absorption of cholesterol, thereby reducing plasma cholesterol in
55

preclinical models of hypercholesterolemia. In rhesus monkeys fed a diet containing

375mg/day of cholesterol, 0.1mg/kg of ezetimibe completely prevented the doubling
of

plasma

cholesterol

normally

(ED50=0.0005mg/kg). Low-density

induced

under

these

dietary

conditions

lipoprotein (LDL) cholesterol was dose

dependently reduced, while high-density lipoprotein (HDL) cholesterol and plasma

triglyceride were unchanged. Clader et al254 synthesized a series of azetidinone
cholesterol absorption inhibitors (CAI) and compounds were evaluated for their
activity to inhibit hepatic cholesteryl ester formation in a cholesterol fed hamster
model. Although originally designed as acyl CoA: cholesteryl acyl transferase
(ACAT) inhibitors , comparison of in vivo potency with in vitro activity in a
microsomal ACAT assay indicated no correlation between activity in these two
models. Examination of the in vivo activity of a range of compounds has revealed
clear structure-activity relationships consistent with a well defined molecular target.
Two derivatives, of a novel cholesterol absorption inhibitors were glucuronidated
with the help of glucuronyl transferase derived from bovine and dog liver
microsomes. An efficient procedure for the iodination was developed on an analytical
scale to be used for the preparation of a radioactive255 glucuronide. Different
authors256-258 have reported the Beta-lactam derivatives as cholesterol lowering
agents.

2.2.6. Beta-lactam derivatives as human tryptase & chymase

inhibiters.
Sutton et al259 prepared a series of non guanidine N1-activated C4-carboxy
azetidinone tryptase inhibitors by solid phase methodology to quickly assess the SAR
associated with the distal functionality on the N1-activating group . From these
studies , potent inhibitors with improved specificity were discovered. Qian et al260
synthesized a highly stereo selective novel tryptase inhibitor. Key to this synthesis
was the discovery and development of a high diastereo selective demethoxy
carbonylation of diester to form the trans-azetidinone. Derivatives of 3benzylazetidine-2-one were designed and evaluated as a novel series of chymase
inhibitors by Aoyama et al261. Structure activity relationship of 3-benzylazetidine-2ones led to compounds, which exhibited 3.1nm inhibition of human chymase and
enhancement of stability in human plasma (t1/2=6hrs). Bisacchi et al262 synthesized a
number of potent azetidinone tryptase inhibitors in which the guanidine moiety at the
56

ring C-3 position is replaced with primary or secondary amine or amino pyridine
functionality . These compounds were found to be highly potent tryptase inhibitors,
which has excellent selectivity against trypsin and most other related serine proteases.
Different authors263,264 have reported the Beta-lactam derivatives as human tryptase
& chymase inhibiters.

2.2.7. Beta-lactam derivatives as anti-hepatitis agents.

Hepatitis A virus (HAV) 3C enzyme is a picornaviral cysteine proteinase
involved in the processing of the initially synthesized viral poly protein is therefore
important for viral maturation and infectivity. Although it is a cysteine proteinase, this
enzyme has a topology similar to those of the chymotrypsin like serine proteinases.
Since the enzyme recognizes peptide substrates with a glutamine residue at the P (1)
site , a number of ketone-containing glutamine compounds analogous to nano molar
inhibitors of cathepsin k were synthesized by Ramtohul et al265 and tested for
inhibition against HAV 3C proteinase. In addition, a 3-azetidinone scaffold was
incorporated into the glutamine fragment but gave only modest inhibition . However,
introduction of a phthalhydrazido group alpha to ketone moiety gave significant better
inhibitors with IC50 values ranging from 13 to 104 m, presumably due to the effect
of intra molecular hydrogen bonding to the ketone. Lall et al266 synthesized a number
of serine and threonine beta-lactones and were tested against HAV 3C proteinase. The
D-N-Cbz-serine beta-lactones displayed competitive reversible inhibition with a K(i)
value of 1.50 10-6 M. Its enantiomer, L-N-Cbz-serine beta-lactone is an irreversible
inactivator with K(inact) = 0.07min-1 , K(lota) = 1.84 10-4 M and K(inact) / K(lota)
= 3800 M-1min-1 . Mass spectrometry and HMQC NMR studies using 13C-labelled LN-Cbz-serine beta-lactone showed that inactivation of the enzyme occurs by
neucleophilic attack

of the cysteine thiol (cys-172) at the beta-position of the

oxetanone ring. Although the N-Cbz-serine beta-lactones displayed potent inhibition ,

other related analogues with an N-Cbz side chain, such as the five-membered ring
homoserine gamma-lactones, the four-membered ring beta-lactam, 2-methylene
oxetane, cyclobutanone and 3-azetidinone, failed to give significant inhibition of
HAV 3C proteinase, thus the importance of the beta-lactone ring for binding has been
demonstrated.

57

2.2.8. Beta-lactam derivatives reduces ethanol consumption in

alcohol-preferring rats.
Changes in glutamatergic transmission affect many aspects of neuroplasticity
associated with ethanol and drug addiction. For instance, ethanol and drug seeking
behavior is promoted by increased glutamate transmission in key regions of the
motive circuit. Youssef Sari et al267 hypothesized that because glutamate transporter 1
(GLT1) is responsible for the removal of most extracellular glutamate, up-regulation
or activation of GLT1 would attenuate ethanol consumption. Behavioral drinking,
Statistical analyses revealed a significant reduction in daily ethanol, but not sucrose,
consumption following Ceftriaxone (CEF) treatment. During the post treatment
period, there was a recovery of ethanol intake across days. Dose-dependent increases
in water intake were manifest concurrent with the CEF-induced decreases in ethanol
intake. Nevertheless, CEF did not affect body weight. An examination of a subset of
the CEF-treated ethanol-drinking rats, on the third day post CEF treatment, revealed
increases in GTL1 expression levels within the prefrontal cortex and nucleus
accumbens. These results indicate that CEF effectively reduces ethanol intake,
possibly through activation of GLT1, and may be a potential therapeutic drug for
alcohol addiction treatment.

2.2.9. Beta-lactams decreases acquisition of and motivation to

respond for cocaine, but not sweet food, in mice.
No medication is approved to treat cocaine addiction, but mounting evidence
by Ward et al268 reported that glutamate-directed approaches may reduce cocaine
dependence and relapse. The glutamate transporter subtype 1 activator, ceftriaxone,
disrupts acquisition of cocaine self-administration, motivation to self-administer
cocaine, and conditioned place preference in mice. Repeated ceftriaxone (200 mg/kg)
reduced the ability of mice to acquire cocaine and the motivation to self-administer
cocaine after successful acquisition without affecting acquisition of or motivation for
sweet food. Repeated ceftriaxone had no effect on cocaine-conditioned place
preference. These results suggest that a -lactam antibiotic reduces the direct
reinforcing strength of cocaine without producing nonspecific deficits in conditioned
learning processes.
58

2.2.10. Artesunate enhances the antibacterial effect of beta-lactam

derivatives.
Yao et al

269

suggested that although artesunate itself had no anti bacterial

ability, artesunate significantly increased the antibacterial effect of -lactam

antibiotics against E. coli ATCC 35218. Artesunate increased daunomycin
accumulation within E. coli in a dose-dependent manner and reduced the mRNA
expression of AcrAB-TolC, an important multidrug efflux system for Gram-negative
bacteria. The bacterial number was significantly reduced by as-ODN targeting AcrB,
but did not further decrease after additional artesunate treatment. In contrast,
artesunate lost its enhancement of -lactam antibiotics against E. coli AG100A, a
strain lacking the gene encoding AcrAB.

2.2.11. Genetic determinants involved in the susceptibility of

Pseudomonas aeruginosa to beta-lactam antibiotics.
Alvarez-Ortega et al270 suggested that the resistome of P. aeruginosa for three
-lactam antibiotics, namely, ceftazidime, imipenem, and meropenem, was
deciphered by screening a comprehensive PA14 mutant library for mutants with
increased or reduced susceptibility to these antimicrobials. Confirmation of the
phenotypes of all selected mutants was performed by E-test. of the total of 78
confirmed mutants, 41 demonstrated a reduced susceptibility phenotype and 37 a
super susceptibility (i.e., altered intrinsic resistance) phenotype, with 6 mutants
demonstrating a mixed phenotype, depending on the antibiotic. Only three mutants
demonstrated reduced (PA0908) or increased (glnK and ftsK) susceptibility to all
three antibiotics. Overall, the mutant profiles of susceptibility suggested distinct
mechanisms of action and resistance for the three antibiotics despite their similar
structures. More detailed analysis indicated important roles for novel and known lactamase regulatory genes, for genes with likely involvement in barrier function, and
for a range of regulators of alginate biosynthesis.

2.2.12. Allergy to beta-lactam antibiotics

Ponvert et al271 suggested, studies based on skin and challenge tests have
shown that 1260% of children with suspected beta-lactam hypersensitivity were

59

allergic to beta-lactams. Responses in skin and challenge tests were studied in 1865
children with suspected beta-lactam allergy272 (i) to confirm or rule out the suspected
diagnosis; (ii) to evaluate diagnostic value of immediate and non-immediate responses
in skin and challenge tests; (iii) to determine frequency of beta-lactam allergy in those
children, and (iv) to determine potential risk factors for beta-lactam allergy. The
work-up was completed in 1431 children, of whom 227 (15.9%) were diagnosed
allergic to beta-lactams. Beta-lactam hypersensitivity was diagnosed in 50 of the 162
(30.9%) children reporting immediate reactions and in 177 of the 1087 (16.7%)
children reporting non-immediate reactions (p < 0.001). The likelihood of beta-lactam
hypersensitivity was also significantly higher in children reporting anaphylaxis, serum
sickness-like reactions, and (potentially) severe skin reactions such as acute
generalized exanthematic pustulosis, StevensJohnson syndrome, and drug reaction
with systemic symptoms than in other children (p < 0.001). Skin tests diagnosed 86%
of immediate and 31.6% of non-immediate sensitizations. Cross-reactivity and/or
cosensitization among beta-lactams was diagnosed in 76% and 14.7% of the children
with immediate and non-immediate hypersensitivity, respectively. The number of
children diagnosed allergic to beta-lactams decreased with time between the reaction
and the work-up, probably because the majority of children with severe and worrying
reactions were referred for allergological work-up more promptly than the other
children. Sex, age, and atopy were not risk factors for beta-lactam hypersensitivity. In
conclusion, it is confirmed in numerous children that (i) only a few children with
suspected beta-lactam hypersensitivity are allergic to beta-lactams; (ii) the likelihood
of beta-lactam allergy increases with earliness and/or severity of the reactions; (iii)
although non-immediate-reading skin tests (intradermal and patch tests) may diagnose
non-immediate sensitizations in children with non-immediate reactions to betalactams (maculopapular rashes and potentially severe skin reactions especially), the
diagnostic value of non-immediate-reading skin tests is far lower than the diagnostic
value of immediate-reading skin tests, most non-immediate sensitizations to betalactams being diagnosed by means of challenge tests; (iv) cross-reactivity and/or
cosensitizations among beta-lactams are much more frequent in children reporting
immediate and/or anaphylactic reactions than in the other children; (v) age, sex and
personal atopy are not significant risk factors for beta-lactam hypersensitivity; and
(vi) the number of children with diagnosed allergy to beta-lactams (of the immediatetype hypersensitivity especially) decreases with time between the reaction and
60

allergological work-up. Immunologic cross-reactivity of aztreonam with other betalactam antibiotics has been studied by Saxon et al273 and Rodilla et al274.

2.2.13. Integrated detection of extended-spectrum beta-lactam

resistance.
Different ways of bacterial resistance278-286 have been shown in (Figure No.
3), Leinberger et al275 suggested that extended-spectrum beta-lactamases (ESBL) of
the TEM, SHV, or CTX-M type confer resistance to beta-lactam antibiotics in Gramnegative bacteria. The activity of these enzymes against beta-lactam antibiotics and
their resistance276,277 against inhibitors can be influenced by genetic variation at the
single-nucleotide level.He suggested the development and validation of an
oligonucleotide microarray for the rapid identification of ESBLs in Gram-negative
bacteria by simultaneously genotyping blaTEM, blaSHV, and blaCTX-M. The array
consists of 618 probes that cover mutations responsible for 156 amino acid
substitutions. As this comprises unprecedented genotyping coverage, the ESBL array
has a high potential for epidemiological studies and infection control. With an assay
time of 5 h, the ESBL microarray also could be an attractive option for the
development of rapid antimicrobial resistance tests in the future. The validity of the
DNA microarray was demonstrated with 60 blinded clinical isolates, which were
collected during clinical routines. Fifty-eight of them were characterized
phenotypically as ESBL producers. The chip was characterized with regard to its
resolution, phenotype-genotype correlation, and ability to resolve mixed genotypes.
ESBL phenotypes could be correctly ascribed to ESBL variants of blaCTX-M (76%),
blaSHV (22%), or both (2%), whereas no ESBL variant of blaTEM was found. The most
prevalent ESBLs identified were CTX-M-15 (57%) and SHV-12 (18%).

61

(Figure No. 3) Mechanism of bacterial resistance

2.2.14. Beta-lactam antibiotic inhibits development of morphine

physical dependence in rats.
Rawls et al287 suggested that -Lactam antibiotics enhance cellular glutamate
uptake. As increased glutamatergic transmission is a primary mediator of opiate
dependence, tested the hypothesis that a -lactam antibiotic (ceftriaxone) prevents
development of morphine physical dependence in rats. Morphine (20mg/kg) was
injected twice daily for 10 days to induce physical dependence. Naloxone (10mg/kg)
administration 1, 48, and 96h after the last morphine injection induced a withdrawal
syndrome characterized by the appearance of wet-dog shakes, teeth chattering, eye
blinking, jumping, and paw tremor. Ceftriaxone (150, 200mg/kg) injected once daily
during chronic morphine exposure inhibited each naloxone-precipitated withdrawal
sign. Ceftriaxone efficacy persisted even after the 96h-naloxone (10mg/kg) injection.
These results suggest that -lactam antibiotics inhibit processes leading to
development of morphine physical dependence.

62

2.2.15. Synergistic antibacterial effect of Beta-lactam derivatives

with silver nano-particles.
Ping et al288 The bactericidal action of silver nanoparticles and amoxicillin on
Escherichia coli is studied, respectively. Increasing concentration of both amoxicillin
(00.525 mg ml1) and silver nanoparticles (040 g ml1) showed a higher
antibacterial effect. Escherichia coli cells have different bactericidal sensitivity to
them. When amoxicillin and silver nanoparticles are combined, it results in greater
bactericidal efficiency on Escherichia coli cells than when they were applied
separately.

2.2.16. Beta-lactam derivatives offer neuroprotection.

Glutamate is the principle excitatory neurotransmitter in nervous system.
Inactivation of synaptic glutamate is handled by glutamate transporter (GLT1).
Animal studies show that the GLT1 is important for normal excitatory synaptic
transmission, while its dysfuction is implicated in acute and chronic neurological
disorders including stroke, brain tumours and epilepsy. It has been discovered that
may -lactam antibiotics are potent stimulators of GLT1,Glutamate transporter are
important in preventing glutamate neurotoxicity289.

2.2.17. Bacteriological antagonism between acylureidopenicillins

& cephalosporins, and combination of different antibiotics.
An antagonism is described by Grimm290 between cefoxitin and azlocillin by
means of agar-diffusion test and checker-board titrations of MIC. This phenomenon is
attributed to beta-lactamase-induction by cefoxitin. Cefuroxime is less antagonistic,
and cefotaxime is indifferent in combination with azlocillin. Combination of
mezlocillin and azlocillin with cephalosporin antibiotics has been studied by Neu et
al291 for their synergistic effects. Amdinocillin in combination with another betalactam antibiotic (ampicillin, cephalothin, cefamandole or cefoxitin) was studied by
Rosten et al292.

63