Curr. Med. Chem.

- Anti-Cancer Agents, 2003, 3, 439-450 439

The Quinolone Family: From Antibacterial to Anticancer Agents

Claudia Sissi* and Manlio Palumbo

Dept. of Pharmaceutical Sciences, V. Marzolo 5, 35100 Padova, Italy

Abstract: The present review focuses on the structural modifications responsible for the transformation of an antibacterial
into an anticancer agent. Indeed, a distinctive feature of drugs based on the quinolone structure is their remarkable ability
to target different type II topoisomerase enzymes. In particular, some congeners of this drug family display high activity
not only against bacterial topoisomerases, but also against eukaryotic topoisomerases and are toxic to cultured mammalian
cells and in vivo tumor models. Hence, these cytotoxic quinolones represent an exploitable source of new anticancer
agents, which might also help addressing side-toxicity and resistance phenomena. Their ability to bind metal ion co-
factors represents an additional means of modulating their pharmacological response(s). Moreover, quinolones link
antibacterial and anticancer chemotherapy together and provide an opportunity to clarify drug mechanism across divergent
species.

THE QUINOLONE FAMILY: FROM ANTIBAC-
TERIAL TO ANTICANCER AGENTS
The history of quinolones begins in the early 1960’s with
the synthesis of the first member of the family, nalidixic acid
(Fig. 1) [1]. Its antibacterial properties appeared immediately
to be very interesting. Unfortunately, its applications in
humans revealed only a moderate response and it was soon
dropped from clinical use. However, the generally simple
and flexible synthetic route to quinolone derivatives, allowed
to producing a large number of analogs including the most
active and broad spectrum oral antibacterials currently in use
[2].
A most successful achievement was the preparation of
fluoroquinolones. In fact, the introduction of a fluorine atom
at the C-6 position increased several folds the activity of the
drug and led to compounds active against a broader spectrum
of bacteria. The first fluoroquinolone clinically used was
norfloxacin (Fig. 1). Its poor tissue distribution limited its
applications to the treatment of urinary tract infections.
Modification of the substituent at N-1 position produced
ciprofloxacin (Fig. 1). This drug presents an excellent
systemic activity upon oral administration and has become
indeed one of the most frequently prescribed antibiotics.
More recently, fluoroquinolones have been further optimised
to improve both pharmacokinetic and pharmacodynamic
properties like favorable bioavailability allowing oral
administration, good tolerability, high tissue concentrations
as well as superior bactericidal activity against a broad
spectrum of clinically relevant pathogens including
enterobacteria, Pseudomonas aeruginosa, Staphylococcus
aureus and Streptococcus pneumoniae. Thus, new
generations of fluoroquinolones are now available [2]. They
preserve excellent potency against Gram-negative bacteria
and, at the same time, they show an increased activity
towards Gram-positive bacteria compared to ciprofloxacin.
*Address correspondence to this author at the Dept. of Pharmaceutical
Sciences, V. Marzolo 5, 35100 Padova, Italy; Tel: +39-049-827-5717;
Fax: +39-049-827-5711; E-mail: claudia.sissi@unipd.it
At the moment, the main interest for these compounds is
related to the treatment of respiratory diseases [3].
O O
O
H3 C N N
NALIDIXIC ACID
O O O O
F F
O O
N N N N
HN HN
NORFLOXACIN CIPROFLOXACIN
Fig. (1). Chemical structure of antibacterial quinolones.
Unfortunately, the high efficiency and broad spectrum of
activity shown by quinolones are now seriously challenged
by the selection of resistant strains, even for naturally highly
susceptible species like Escherichia coli [4]. Thus, the
present research in the field is specially devoted to overcome
resistance phenomena that are emerging as the most relevant
complication in antibacterial therapies.
Biological Targets of Antibacterial Quinolones
Forty years of research in the quinolone field have
allowed to elucidating some key aspects of their mechanism
of action. Their main biological targets are the DNA gyrase
(Gyr) and topoisomerase IV (Top4), two enzymes belonging
to the type II topoisomerase family [5, 6]. These enzymes are
essential in all bacteria and are expressed in prokaryotic
organisms only, features that make them ideal antibacterial
drug targets. Indeed, the prominent selectivity of quinolones

1568-0118/03 $41.00+.00 © 2003 Bentham Science Publishers Ltd.

440 Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6
in poisoning the bacterial type II topoisomerases vs. the
corresponding mammalian enzymes represents the rational
basis for their safe use in antibacterial chemotherapy.
Gyr and Top4 exhibit close structural and functional
similarities. This emerges from their high level of primary
sequence homology and from their common catalytic cycle.
In fact, both cleave two DNA strands in a concerted
sequence of events and transport one duplex DNA segment
through the break. In so doing, they can catenate or
decatenate duplex DNA rings and alter DNA topology. The
major difference between the two enzymes rests on the fact
that Gyr is able to introduce negative supercoils into DNA,
whereas Top4 is not.
Gyr is formed by two different subunits, GyrA and GyrB,
that are arranged in a A2B2 tetramer to constitute the active
protein [7, 8].
GyrA is a 97 kDa protein (875 aa) encoded by gyrA.
Tryptic cleavage of this subunit generates two fragments that
correspond to two main functional domains. The 64kDa N-
terminal domain (residues 7 to 571) contains the catalytic
tyrosine (Tyr122), whereas the 33kDa C-terminal domain
(residues 572 to 875) is involved in DNA binding and
wrapping.
GyrB is an 89.9 kDa (804 aa) protein encoded by the
gyrB gene. Also this protein contains two functional
domains. The 43kDa N-terminal fragment (aa 2-393) is the
ATP binding and hydrolysis domain; the 47 kDa C-terminal
domain (aa 394-804) is involved in interactions with GyrA
and DNA.
More recently, the identification of the genes encoding
Top4 subunits, parC and parE, confirmed a similar
quaternary structure for the two prokaryotic enzymes [9,10].
In fact parC encoded an 83.7 kDa protein of 752 aa
called ParC, whereas parE encoded a 70.2 kDa protein of
630 aa called ParE [10, 11]. These two proteins share an
extended sequence homology with GyrA and GyrB
respectively and a similar distribution of functional domains
[11,12]. In addition, the Top4 active form is constituted by a
C2E2 tetramer as in the case of Gyr.
The catalytic cycle of the two enzymes is essentially the
same. They initially bind a dsDNA segment (G segment).
Complex formation with ATP induces the closure of an
ATP-operated clamp formed by the N-terminal domains of
the GyrB or ParE dimer that lead to the capture of a second
dsDNA fragment, the T segment [13]. Thus, a concerted
mechanism occurs: trapping of the T segment induces
cleavage of the G segment on both strands. In the cleavage
complex a tyrosine is covalently bound to the 5’-end of the
DNA cleavage site. On the opposite strand, the cut is 4-bp
staggered [14]. The combination of these events triggers
nucleotide hydrolysis. ADP rapidly dissociates from the
proteins, thus opening the clamp and allowing the release of
the T-segment after its crossing through the G-gate. The re-
sealing and release of the G-segment allows the enzyme to
start a new catalytic cycle [15].
Despite the extensive similarity in sequence, structure
and functional properties, the two topoisomerases have
distinct identities and roles.
Sissi and Palumbo
In vitro, the two enzymes discriminate selectively
between intra and inter-molecular topological changes: while
Top4 catalyses, preferentially intermolecular reactions like
decatenation, Gyr favors an intramolecular reaction such as
supercoiling [16].
These differences are particularly significant in vivo. In
fact, although both enzymes are essential for the proper
occurrence of chromosome duplication and segregation
processes, they probably work at different stages of
replication. In particular, experimental evidence suggests
that Top4 prefers to unlink precatenane, whereas DNA
gyrase works to remove positive supercoils [17-19].
Mechanism of Action of Antibacterial Quinolones
Quinolones act in a similar fashion against the two
prokaryotic type II enzymes. They essentially block the
topoisomerase catalytic cycle when the protein is covalently
linked to the cleaved DNA (cleavage complex). Stabilization
of the enzyme-DNA complex has been confirmed to block
the progression of the replicative machinery and to create
DNA lesions that induce bacterial SOS response [20-22].
At the molecular level, the interactions of quinolones
occur mainly with the A subunit of Gyr and with ParC of
Top4. In fact, a region spatially close to the active tyrosine
and called Quinolone Resistance Determining Region
(QRDR) has been identified. Aminoacid mutations in this
region greatly reduce the affinity of quinolones for the
enzyme-DNA complex. The most relevant positions in E.
Coli are Ser83 and Asp87 in GyrA, and the corresponding
Ser79 and Asp83 in ParC [23, 24]. Mutations in GyrB and
ParE, although implicated in fluoroquinolone resistance, are
less frequent. The current hypothesis is that they occur at the
interface between the two subunits inducing subtle structural
changes in the overall protein architecture.
The structural features of the quinolone-gyrase-DNA
complex have been only partially elucidated [25].
Experimental evidence suggests that quinolones, in the
presence of proper metal ions, bind single stranded DNA and
GyrA with moderate affinity [26, 27]. However, they bind
efficiently to the gyrase-DNA complex [28].
At the DNA level, cleavage is not required to stimulate
drug binding: the structural distortion of the double helix
bound to the protein and stabilized by selective contacts with
protein residues, appears to be sufficient to provide a
favorable interaction site for quinolones [29]. Additionally,
the presence of quinolones alters not only the structural
features of GyrA (binding site) but also those of the overall
A2B2-DNA complex [27, 30] inducing modifications in the
kinetic rates of different catalytic steps.
Role of Quinolones in Eukaryotic Cells
In eukaryotes, DNA gyrase and topoisomerase IV are
functionally replaced by two isoenzymes: topoisomerase II α
and β, a 170 and a 180 kDa protein respectively [5]. These
proteins share a similar catalytic cycle, the main difference
resting in the fact that the active form of eukaryotic
topoisomerase II (Top2) is constituted by a homodimer.
However, as supported by the extended sequence homology,
Top2 can be seen as the fusion of the GyrA-GyrB or ParC-
The Quinolone Family
ParE subunits. Additionally, although no crystallographic
data are available at present for any of the above mentioned
full length enzymes, the structures of the crystallized
fragments of GyrB and GyrA have been found to be
complementary to the corresponding fragments of yeast
Top2 [31-33].
Like bacterial Gyr and Top4, also eukaryotic Top2 is a
validated pharmacological target. In fact, many clinically
useful anticancer drugs exert their cytotoxic action by
interfering with this enzyme. Such drugs can be divided into
two main classes: inhibitors and poisons.
Topoisomerase inhibitors, by binding to the enzyme,
prevent it from performing its catalytic cycle. This impairs
DNA processing and causes cytostatic effects. As it is highly
preferable to kill a cancer cell than to stop its growth, Top2
inhibitors are not particularly appealing as drug candidates.
On the other side, Top2 poisons act in the same way as
quinolones: by increasing the DNA cleavage rate or by redu-
cing the DNA religation rate, the so-called cleavage complex
spans its life and DNA breaks persist longer. Stabilization of
the cleavage complex converts the topoisomerase into an
endogenous toxin that “freezes” DNA damage in the
genome. This triggers a sequence of biochemical events that
finally leads to a programmed cell death, called apoptosis.
Common features in the mechanism of action of antibac-
terial quinolones and antitumor drugs have suggested that
both compounds have a similar mode of interaction with the
type II DNA topoisomerase-DNA complexes. In particular,
comparison between quinolones and antitumor drugs of the
epipodophyllotoxin family (etoposide and teniposide) has
been considered especially relevant, since neither type of
drug is a DNA intercalator [34]. Further indications came
from studies carried out within archaebacteria. These
organisms occupy an intermediate phylogenetic position
between eukaryotes and eubacteria [35]. In these systems,
comparable sensitivity to poisons of each type II topoiso-
merase was observed and the DNA cleavage patterns induced
by ciprofloxacin and etoposide were found to be very similar
[36, 37]. These findings strongly supported a common mode
of interaction with the DNA-DNA topoisomerase II com-
plexes for ciprofloxacin and etoposide.
How can a small molecule discriminate efficiently
between remarkably similar enzymes? Due to the lack of
information concerning the cleavage complex in the
presence/absence of drugs, no final answer is available at the
moment. However, this open question gives a new input in
the field: use quinolones as leading compounds to be
optimized not against their “traditional” targets (Gyr/Top4)
but towards mammalian Top2, thus transforming antibac-
terial into anticancer drugs.
One of the first reports in this direction came in the
middle 80’s, when the influence of some antibacterial
quinolones on elements of the eukaryotic DNA replication
machinery was tested in vitro [38]. It was found that
quinolones such as ciprofloxacin, norfloxacin and ofloxacin
interfered only slightly with the function of calf thymus
Top2, the prokaryotic enzyme Gyr being approximately 100-
fold more sensitive to inhibition than its eukaryotic
counterpart. Starting from this pioneering study, systematic
Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6 441
investigations on quinolone selectivity between bacterial and
mammalian type II topoisomerases have been carried out
[39-41]. It emerged that, generally, the inhibitory activities
of quinolones against prokaryotic or eukaryotic type II
topoisomerases significantly correlated with their
antimicrobial or cytotoxic activities respectively. However,
no relationship has been established between the two
mechanisms as large differences in selectivity were observed
on varying the nature of the test quinolones.
In the late eighties, quinolones having a 4-pyridinyl
group at C-7 were synthesized. Among them rosoxacin
gained interest as it was particularly effective in treating
gonococcal infections (Fig. 2) [42]. With the aim to increase
the quinolone activity toward gram-positive bacteria various
7-(4-pyridinyl) derivatives were examined [43]. Remarkably,
some of these new compounds produced an unacceptable
toxicity profile in vitro inducing micronuclei, a genetic
toxicity endpoint.
At the same time, a new 6,8-difluoro-7-pyridyl-4-
quinolone, CP-67, 015, with potent antibacterial activity, was
shown to strongly enhance topoisomerase II-mediated DNA
cleavage, which was related to its reported clastogenic effect
on DNA in cell culture and to its positive mutagenic
response in mouse lymphoma cells [44].
Since then, different directions were followed to eluci-
date the structure activity relationships responsible for the
interference of quinolones with eukaryotic topoisomerase II.
Cytotoxic Quinolones: Structure Activity Relationships
Initial approaches to define proper SAR for cytotoxic
quinolones were based on systematic structural modifica-
tions of canonical antibacterial fluoroquinolones. These lead
to the identification of characteristic structural features
linked to cytotoxic activity against eukaryotic cells. In parti-
cular, starting from norfloxacin or ciprofloxacin, it emerged
that no single position has a controlling effect on the
observed cytotoxicity but, more likely, a combination of the
various substituents contributes to directing the poisoning
activity. A schematic evolution of cytotoxic quinolones is
reported in Fig. (2).
The presence of a fluorine atom at position C-6 of the
quinolone ring is a general requirement to grant potent
antibacterial activity in “classical” quinolones [45]. It has
been proved that, in order to stabilize the cleavage complex
with mammalian Top2, the presence of two halogens at C-6
and C-8 of quinolone skeleton is preferred. In fact, a direct
comparison of 6-fluoro (CP-115, 955) and 6,8-difluoro (CP-
115, 953) derivatives showed that removal of the C-8
fluorine decreased the ability of the quinolone to stimulate
enzyme-mediated DNA cleavage by approximately 2.5-fold.
Activity on the mammalian enzyme parallels the cell killing
ability observed towards Chinese hamster ovary cells, confir-
ming that the C-8 fluorine increases the potency of quinolone
derivatives against Top2 and mammalian cells [46].
In the presence of fluorine atoms at 6 and 8 positions, the
change of N-1 substituent from ethyl to cyclopropyl was also
critical for antiproliferative activity. In fact, in most
homogeneous series of derivatives, the presence of a
cyclopropyl group increased the activity compared to an
442 Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6 Sissi and Palumbo

O O

N
N
ROSOXACIN

O O O O
F F
O O
H3 C
N
N F N F

CP-67,015 CH3
WIN 57294

O O O

F F H
O
H3 C
N R

F N F
HO
CH3
CP-67,804 R = H, CH 3, C6H5

O O O OH
F F
O
H3 C
HO
F N F
HO

CP-115,953 CH3
WIN 64593

Fig. (2). Historical development of some cytotoxic quinolones.

ethyl group [47, 48]. Thus, it seems that at N-1, the SAR
profile obtained for antibacterial activity is conserved for cell
cytotoxicity. Combining the above findings, it is suggested
that the concerted presence of the halogen at C-8 and the
cyclopropyl at N-1 may produce better interference of
quinolones with Top2 than with Gyr [49, 50].
A variety of substitutions were also introduced at
position C-7. In fact, the presence of a basic amino group in
the aliphatic cyclic substituent at position C-7 is a strict
requirement for antibacterial activity. Additionally,
modifications at this position where shown to play a key role
in directing the drug preferentially towards Gyr or Top4
[51]. These results suggested a direct interaction of this
portion of the quinolone with the enzyme. Conceivably, the
C-7 substituent should affect the prokaryotic/eukaryotic
enzyme selectively. Indeed, methyl substituents at the C-7
piperazine group influenced potency against the mammalian
enzyme, stereochemistry being a discriminating factor. In
fact 3,5-dimethylpiperazinyl derivatives were active in
stimulating enzyme-mediated DNA cleavage only in the
trans configuration. This was particularly interesting because
cis- or trans-methyl substitution on the piperazine had little
effect on the activity against Gyr, suggesting that only in the
mammalian enzyme an asymmetric barrier exists which
The Quinolone Family
influences productive quinolone interaction and favors the
less bulky trans-3,5-dimethylpiperazine substituent at C-7
[52].
Parallel studies evidenced that compounds with C-7
pyrrolidine substituents were more cytotoxic than those with
piperazine substituents [50]. This is actually in line with
observations made on derivatives related to the earlier
cytotoxic quinolones [44]. Looking both at the stimulation of
DNA cleavage and at the cytotoxicity against cultured
mammalian cells, it was observed that the presence of an
aromatic group contributed greatly to drug activity. In
particular, in this series of compounds, a 4'-hydroxyphenyl
substituent at the C-7 position was critical for potency
towards the mammalian Top2 [48]. The compendium of
these studies on structure-activity relationships is represented
by CP-115, 953, [6,8-difluoro-7-(4'-hydroxyphenyl)-1-
cyclopropyl-4- quinolone-3-carboxylic acid], one of the most
cytotoxic quinolone thus far synthesized (Fig. 2) [53].
More recently, novel substitutions were introduced at
position C-7, which can provide good models for further
design of potent antitumor quinolones. A remarkable example
is the introduction of an unsaturated aminoazabicyclo group
[47]. Compounds of this type are peculiar because they
associate maximal activity with the presence of an atypical
amino group at C-5 of the quinoline carboxylic acid. The
possibility to substitute the 5-H with NH2 or F, maintaining
the activity against mammalian Top2 was further confirmed
in other series of derivatives [52, 54].
In the absence of an aromatic moiety at position C-7, a 7-
[4-(2-hydroxyiminoethyl)piperazin-1-yl] group has been
introduced. Among various combinations, 1-ethyl-6-fluoro-7-
{4-[2-(4-chlorophenyl)-2-hydroxyiminoethyl]-1-piperazinyl}-
4-oxo-1,4-dihydro-3-quinoline carboxylic acid demonstrated
the most significant activity against renal cancer cell lines
[55].
Other intriguing results concern position C-3. It is well
established that, in antibacterial quinolones, the 3-COOH or
its isosteric replacement is a fundamental requirement for
activity (Fig. 2). However, this functional group can be
replaced even with an hydrogen still granting effective
poisoning of Top2. It seems that, to inhibit the eukaryotic
enzyme, the basic requirement is the coplanarity of the C-3
substituent with the quinoline ring. Thus, although in 3-H
derivatives small substituents can be beneficial at C-2, no
such residue can be introduced if the carboxyl group is
present, as it will destabilize the coplanar orientation of the
acidic moiety [54].
Removal of the carboxylic group opened up new
synthetic opportunities. In fact, introduction of a phenyl
group or a related heteroaromatic ring at C-2 enhanced Top2
poisoning activity [56]. The distance between the two
aromatic moieties is apparently crucial, as only a methylene
linker allowed to maintain biological activity. Further
increase in cytotoxic activity was obtained upon introduction
of hydroxyl- substituents into the C-2 phenyl group. In
particular, the 2,6-dihydroxy benzyl derivative (Win 64593)
is the most active compound available to date. The role of
hydroxyl substitution is not clear yet. In quinolones carrying
the carboxyl group, the acidic moiety can participate with the
ketone at position 4 in chelating metal ions or can be
Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6 443
involved in hydrogen bonding with the target macromole-
cules. In the new derivatives, the profound modifications
introduced suggest the onset of new interactions involving
the drug aromatic and/or hydroxyl-groups in the cleavage
complex.
In the 7-(2,6-dimethyl-4-pyridinyl) family, the substitu-
tion of the nitrogen at position 1 with a sulphur atom leads to
inactive derivatives [54]. However, as reported in Fig. (3),
other positions can support the introduction of this heteroa-
tom without the loss of activity. An example is represented
by the isothiazoloquinolones A-65281 and A-65282 [57].
These derivatives exhibit a heterocyclic moiety added to the
quinolone system, which can be considered as a modification
of the carboxylic group. It is worth noting that, in spite of the
fact that these derivatives do not exhibit an aromatic
substituent at position C-7, they are active against both
bacterial and eukaryotic type II topoisomerases.
More recently, hetero substitution produced thieno[3'
2':4.5]thieno[2,3-c]quinolones [58]. In this series, derivatives
bearing a 3-dimethylaminopropyl substituent on the quinolone
nitrogen and a methoxycarbonyl group at position 9 exhi-
bited prominent antitumor activity. Cytotoxicity was reduced
when an anilido substituent was present at position 9.
A similar approach leads to the screening of 1,8-
naphthyridine derivatives. Indeed, a series of 1,7-
disubstituted-6-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine-
3-carboxylic acids with cytotoxic activity comparable to that
of etoposide on cancer cell lines in vitro and on murine P388
leukemia in vivo has been very recently disclosed, the
leading compound being AT 3639-1 (Fig. 3) [59]. SAR
studies performed by changing N-1 and C-7 positions and
the core ring structure led to the conclusion that a
heterocyclic 2-thiazolyl group at the N-1 position of the
naphthyridine structure is the best substituent for antitumor
activity. Regarding core ring structure, the naphthyridine
derivative, reminding nalidixic acid, is the most active
followed by pyridopyrimidine analogs. Finally, aminopyrro-
lidine substituents at the C-7 position were more effective
than other amines or thioethers.
Other cytotoxic derivatives are based upon the ofloxacin
or pyrimido[1,6-a]benzimidazole skeleton and thus they
present an additional ring with reference to classical
quinolones. Examples are WIN 58161 and Ro 47-3359
respectively, reported in Fig. (4). The WIN derivative was
identified as a topoisomerase II poison during the screening
of several enantiomerically pure (2,6-dimethyl-4-pyridinyl)
quinolones, previously shown to be potent inhibitors of
bacterial DNA gyrase. Among these and other analogs,
topoisomerase II inhibitory potency was found to depend on
the size and substitution of the bridge spanning the 1- and 8-
positions of the quinoline ring [60]. The use of the
enantiomerically pure WIN 58161 confirms that Gyr and
Top2 inhibitory activities are enantioselective processes. In
fact only the (S) enantiomers show biological activity [61].
The pyrimido[1,6-a]benzimidazole family is another
class of drugs modelled on quinolones and generally active
against DNA gyrase [62]. To introduce cytotoxic activity,
the aliphatic group (4-N-methylpiperazine) at the ring
position equivalent to C-7 in quinolones was again replaced
444 Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6 Sissi and Palumbo

O O
F
NH
N N S

N F
O O
F A 65281

NH O O
R X N S F
R NH
isothiazoloquinolones N N S
H2N
F

A 65282
O O
F
O

N N N
H2N

N S

AT 3639-1

Fig. (3). Chemical structure of cytotoxic quinolones containing a sulphur atom.

O O O O
F F
O O
H3 C
N N N
N O N S CH3
H3 C
H
OFLOXACIN CH3
WIN 58161

OH O OH
O
N N
O F N O
F N
H3 C
N
N N
N N
H3 C
Ro 46-2825 CH3
(CH2) 2N(CH3 )2 Ro 47-3359

N N
F O

H3 C
N
N F

CH3
WIN 63320

Fig. (4). Chemical structure of cytotoxic quinolones containing an extended planar ring system.

with an aromatic basic substituent (2,6-dimethylpyridine) An extended planar ring system is present also in WIN
yielding RO 47-3359 (Fig. 4) [63]. 63320, a pyrazoloquinoline derivative (Fig. 4) [64]. The
The Quinolone Family
pyrazole-ring fusion blocks the carbonyl group in the
conformation previously reported as the active one, which
results in an increment of the in vitro and in vivo cytotoxicity
[54].
Starting from the fact that DNA-intercalating drugs often
exhibit cytotoxic properties, the quinolone structure has been
manipulated in order to yield a more extended planar surface
possibly granting efficient insertion into a double-helical
nucleic acid. A screening program to search for novel
quinolone-related antitumor agents lead to the identification
of the quinobenzoxazine family, characterized by a fused
hetero-tetracycle [65-67]. Several compounds were
synthesized and investigated, some of which exhibited
excellent in vitro cytotoxic activity using cancer cell lines,
including the multidrug resistant P388/ADR line, and also
possessed good activity in vivo against both systemic tumors
and subcutaneously implanted murine solid tumors as well as
human tumor xenografts. The drugs’ potency was typically
in the sub-micromolar range and produced a significantly
increased life span.
Among the tested compounds, two analogs emerged as
particularly interesting, A-62176 (1-(3-aminopyrrolidin-1-
yl)-2-fluoro-4-oxo-4H-quino[2, 3, 4-ij][1, 4]benzoxazine-5-
carboxylic acid) and its amino-analog A-85226, reported in
Fig (5). In both derivatives, the R substituent at position 1 is
an amino-pyrrolidine. Activity was retained and even
increased when introducing an amino group at the 3 position
of the condensed ring system (A-85226).
R5 O the micromolar range.
F COOH
N H
O
H2N
R5 = H A-62176
R5 = NH2 A-85226
Fig. (5). Chemical structure of quinobenzoxazines.
O O
F COOH F COOH
R N R N
O O
1 2
N or
R=
HN
Fig. (6). Chemical structure of pyridobenzophenoxazines.
Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6 445
SAR studies, in particular connecting DNA-binding and
biological activity, were performed for a number of A-62176
analogs [68]. The results indicate that DNA-binding affinity
is predictive for topo II inhibition response. In addition, data
showed the requirement for the carbonyl group at position 4,
for the free amine at position 1 and the possibility of
increasing DNA-binding/topo II activity by introducing
aliphatic (methyl) substitutions at position 9. On the
contrary, fluorine substitution at the latter position resulted in
strong reduction of drug potency.
Finally, considering that the phenyl ring portion of the
quinobenzoxazines might be inserted between DNA base
pairs in the cleavage complex, the drug intercalation ability
was increased by extending the phenyl ring to a naphthyl
ring. A series of new pyridobenzophenoxazine analogs
(compounds 1-3 in Fig. 6) were thus designed and
synthesized [69]. Because of the steric hindrance between
the hydrogens on the pyrido- and benzo- rings, the naphthyl
and quinolone portions of compound 1 are not coplanar. This
may have a negative effect on its DNA intercalation ability.
Therefore, compound 1 was less potent than compounds 2
and 3 and A-62176. Some of the new analogs had a greater
DNA unwinding ability than A-62176, which was apparently
related to higher potency in topoisomerase II inhibition and
increased cytotoxicity against tumor cells.
The tested analogs showed strong cytotoxic effects on
several human and murine cell lines with IC50 values in the
range of 4-2,000 nM. They also showed potent inhibitory
effects against human topoisomerase II, with IC50 values in
Mechanism of Action of Mammalian Topoisomerase II-
Directed Quinolones
The discovery of the poisoning activity of quinolones
against eukaryotic topoisomerase II prompted a relevant
number of studies devoted to clarify the molecular details of
this process. In fact, the drug interactions in the type II
topoisomerase-DNA complex are still ill defined, whereas
the need for such a kind of information is multiple. These
new quinolone derivatives can yield a deep insight of what
occurs in the cleavage complex in the presence of type II
O
F COOH
R N
O
3
N
H2N
446 Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6
topoisomerases of divergent species. A more precise model
for the ternary complex would be useful in designing new
antibacterial derivatives safer for human use on one hand and
new more potent congeners with anticancer activity on the
other hand.
In this connection, CP-115,953 was the most widely
investigated drug. In fact, as an antibacterial, it has the broa-
dest activity spectrum of any other quinolone, being very
effective toward both Gyr and Top4. In addition, it showed
an activity higher than etoposide against any eukaryotic
topoisomerases II examined to date [48, 53, 70-73].
As previously mentioned, the starting point of this
research was a possible similarity between etoposide and
quinolone topoisomerase poison mechanisms suggesting
parallel interactions in the cleavage complex [54]. This
hypothesis was sustained by comparable DNA cleavage
patterns produced by quinolone and etoposide in the
presence of different type II topoisomerases [37, 71].
As expected for topo II poisons, both drugs generate the
typical 4 bases staggered double strand breaks on DNA.
Interestingly, sequence analysis of the drug-stimulated DNA
cleavage showed that all test cytotoxic quinolones display a
preference for C at position –1 associated with a weaker
G preference at +5, this profile being shared by etoposide
[74, 75].
Cross-relations of these consensus sequences and drug
potency performed with several quinolones modified at
positions 3 and 8 suggest possible interactions of specific
substituents on the quinolone derivatives with DNA and/or
topoisomerase. In particular, it is proposed that quinolones
may have at least one DNA-interactive region (involving
position 1 and/or 8) and at least one topoisomerase-
interactive region (at position 3) [75].
Additionally, by in vitro mutagenesis followed by in vivo
selection, it was found that a single point mutation
(His1012Tyr) can confer resistance both to CP-115, 953 and
to etoposide [48, 72]. As determined by DNA cleavage
assays, this mutant was resistant both prior to and following
the DNA strand-passage event. In marked contrast, the
mutant enzyme displayed sensitivity to amsacrine (a DNA
intercalating agent) comparable to the wild type and was
several folds hypersensitive to another intercalator,
ellipticine. At first glance, it might appear that type II
topoisomerases can distinguish only between non-
intercalative and intercalative agents.
However, several lines of evidences indicate a non-
complete identity of binding sites for quinolones and
etoposide. Clear indications came from investigations
performed in the presence of “designed” mutant enzymes.
In E. Coli, high levels of quinolones resistance occur
when Ser83 in GyrA is mutated to Trp or Leu. Introduction
of the corresponding mutation in the human or yeast
topoisomerase II, leads to enzymes that acquire the ability to
discriminate between quinolone and etoposide. In fact, a
Ser740Trp yeast or Ser763Trp human enzyme mutant
become resistant to quinolones but hypersensitive to
etoposide (VP-16) [74, 76-77]. In particular, these purified
mutants show enhanced levels of etoposide-stabilized
Sissi and Palumbo
covalent complex as compared to the wild type enzyme and
a reduced cleavage in the presence of CP-115, 953.
Although, it is not apparent whether tryptophan
intercalation into the DNA double helix or a structural
rearrangement of the enzyme conformation occurs in the
mutant, it is suggested that the Ser-Thr mutation affects
mainly the DNA-protein interactions and that changes at the
enzyme/DNA interface might be critical also for the drug
recognition process. Moreover, as it affects the sensitivity of
quinolone as well as of etoposide, the serine residue is likely
(directly or indirectly) involved in a common (or at least
overlapping) site.
This serine residue is located in a conserved CAP homo-
logy domain (a helix-turn-helix motif), a region involved in
DNA interaction. The importance of this region was further
assessed producing more mutants in Saccharomyces
cerevisiae type II topoisomerases. In particular, studies using
Ser740Trp, Gln743Pro, and Thr744Pro mutants showed that
the α4-helix of this domain is critical for selecting the sites
of DNA cleavage prior to cleavage of the G strand. They
also demonstrated that the solvent-exposed residues in the
α4-helix are important in determining the binding as well as
the activity of quinolones/etoposide against topoisomerase
II-DNA complexes [78].
Studies focused on the single catalytic steps of topoiso-
merase II in the presence of etoposide/CP-115,953 have also
been carried out.
Typical poisons of human Top2 like etoposide or m-
AMSA, stabilize the DNA-protein cleaved complex
primarily by reducing the rate of topoisomerase II–mediated
DNA religation [79]. However, the behavior of quinolones
with reference to eukaryotic Top2 is distinct, as they do not
impair the religation rate of cleaved DNA. In fact, the
quinolone-mediated stabilization of the cleavage complex is
due to increased levels of intermediates formed by an
increment of the enzyme’s forward rate of DNA cleavage
[53]. Nonetheless, etoposide and CP-115,953 were found
mutually exclusive in stimulating cleavage reaction as well
as DNA strand passage [80].
Considering the enzyme-catalysed ATP hydrolysis step,
it emerged that, although neither CP-115,953 nor etoposide
share an interaction domain with novobiocin, etoposide can
moderately inhibit ATPase activity [81]. This latter finding
undercuts the common assumption that DNA cleavage-
enhancing drugs are clustered to the cleavage/religation steps
of topoisomerase II.
Furthermore, data on cell lines are available. In particular,
the VpmR-5 line (an epipodophyllotoxin-resistant Chinese
hamster ovary line) showed cross-resistance to CP-67,804
(approximately 3.7-fold) and CP-115,953 (approximately
1.3-fold). Although quinolone cross-resistance was less
pronounced when compared to etoposide (approximately 12-
fold), this further confirms that topoisomerase II is a
physiological target for CP-67,804 and CP-115,953 in mam-
malian cells [53].
Altogether, these experimental observations led to a
model according to which quinolones and etoposide share
overlapping but not identical site on topoisomerase II [82].
The Quinolone Family
The role of direct DNA-drug interactions in modulating
potency and specificity is another interesting issue. As far as
quinolones, is well establish that they bind efficiently only
single stranded DNA with the assistance of Mg2+ ions. It is to
note that this metal ion is also required by topoisomerase to
perform its catalytic cycle and to bind quinolone. Thus, it
appears that the metal cofactor participates in stabilizing the
drug-cleavage complex interactions. Interestingly, etoposide
shows a similar behavior, since it also binds preferentially
single stranded DNA in the presence of Mg2+ [83].
Since quinobenzoxazines are quinolone derivatives
designed to increment DNA affinity, a number of studies
were focused on their DNA-binding properties [84, 85]. Also
in this case and similarly to their antibacterial congeners,
DNA interaction was found to be dependent upon Mg2+.[26].
It is proposed that the quinobenzoxazine binds duplex DNA
through intercalation in the presence of Mg2+, which bridges
the phosphate backbone of the DNA and the β-ketoacid unit
of the intercalated molecule. A 2:2 drug:Mg2+ complex
appears to form a "heterodimer complex" in which one drug
molecule is intercalated into DNA and the second drug
molecule is externally bound, held to the first molecule by
two Mg2+ bridges, which themselves are chelated to
phosphates on DNA. A cooperativity in binding of the
quinobenzoxazines to DNA, and a 4:4 drug:Mg2+ complex is
proposed, in which the two externally bound molecules from
two different 2:2 dimers via π−π interactions. Further insight
into the metal ion mediated drug-DNA interaction has been
obtained by photochemical studies on A-62176 (Fig. 5). The
quinobenzoxazine undergoes photodecomposition to highly
fluorescent products. DNA accelerates the photochemical
decomposition of A-62176 up to 80-fold. This process
requires Mg2+, duplex DNA, molecular oxygen, and
intercalation of the drug into the DNA duplex. Furthermore,
it was suggested that the precise positioning of the
intercalated molecule, which is modulated by the structure
and stereochemistry of the externally bound molecule, plays
an important role in determining the rate of photoreaction on
DNA. Apparently, the photocleavage reaction occurs by a
free radical mechanism initiated by abstraction of the 4'- and
1'-hydrogens from the DNA minor groove, indicating that
the quinobenzoxazine-drug-Mg2+ complex is located on the
minor groove of the nucleic acid. Photochemical cleavage of
DNA can occur in the presence of antibacterial
fluoroquinolones but not with the same efficiency [86].
An investigation on the ability of the quinobenzoxazines
to interfere with mammalian Top2 showed that they are
potent inhibitors of this enzyme, both in vitro and in vivo
[87]. According to this study, the quinobenzoxazines,
surprisingly, appear to belong to the class of topo II
suppressors that interfere with the catalytic activity of the
enzyme at a step prior to the formation of the cleavage
complex. However, a subsequent study on A-62176, showed
that, at low drug concentrations and pH 6-7, the
quinobenzoxazine enhances the formation of the cleavage
complex at certain sites but inhibits it at high drug
concentrations [88]. Conversely, at another topo II cleavage
site, A-62176 inhibits the cleaved complex formation at both
low and high drug concentrations. Hence, different
mechanisms impairing topo II activity seem to be operating
at different sites. The reason for this uncommon behavior is
Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6 447
not apparent at present. Using a photocleavage assay,
mismatched sequences, and competition experiments
between psorospermin and A-62176, the drug-binding site
where enzyme poisoning occurs has been suggested to be
located on the DNA base pairs between positions +1 and +2
relative to the cleaved phosphodiester bonds. Based upon
these results, the authors suggest that topo II induces
transient structural distortion, possibly unwinding, of the
DNA that is captured or stabilized by the binding of A-
62176 at the topo II cleavage gate.
In conclusion, the quinobenzoxazine represents an
interesting novel pharmacophore with biological properties
distinct from those of antibacterial/cytotoxic quinolones.
A further line of research is based on the intriguing idea
of combining the information obtained using mammalian and
bacterial enzymes.
Competition experiments clearly showed that, indeed, the
drug interaction domain on type II topoisomerases has been
conserved during the evolution from Gyr to Top4 and finally
to Top2.
Indication that similar interactions occur in the two types
of cleavage complex came also from the observation that
ciprofloxacin is actually able to displace both etoposide and
CP-115,953 from their DNA-Top2 cleavage complex, thus
reducing DNA damage. This competitive character is
conserved in vivo as ciprofloxacin inhibited the cytotoxicity
of CP-115,953 and etoposide in mammalian cells to an
extent that paralleled its in vitro attenuation of enzyme-
mediated cleavage [89].
In DNA gyrase, specific residues required to grant
efficient interaction with quinolones have been identified. A
triple mutant of human Top2α in, which three amino acids
have been mutated to the corresponding residues present in
gyrase, Met762Ser, Ser763Ala and Met766Asp showed a
three-fold increase in sensitivity to ciprofloxacin in vitro and
similar sensitivities to a range of other quinolones. On the
other hand, the activity of CP-115,953 was unaffected by any
of these mutations, whereas they conferred resistance to the
non-intercalating agent etoposide [90]. As reported above
and in agreement with these results, also a Thr(744)Pro
substitution in the yeast type II topoisomerase was found to
render the enzyme sensitive to antibacterial quinolones [78].
The above mutations occur in a relatively small region of
topoisomerase. Hence, this region is likely to be involved in
determining the extent of interference of quinolones with the
topoisomerase II-DNA complexes. Thus, the presence of
defined amino acids at positions 762 to 767 appears to
contribute directly (with their side chains) or indirectly
(affecting the enzyme conformation) to create a binding site
for quinolone recognition.
However, it is interesting to point out that subtle changes
are actually occurring from one enzyme to the other in this
overall conserved drug interaction domain. In fact, in
prokaryotic type II topoisomerases (with maximal evidence
in gram positive Top4), the stabilization of the cleavage
complex by quinolones is insensitive to the presence of an
aromatic moiety at position C-7 and is produced by an
inhibition of the religation rate. On the opposite, quinolones
act against Top2 by accelerating the rate of DNA cleavage
448 Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6
and require aromatic substituents at C-7 [71]. Nonetheless,
they operate against the sensitive Thr(744)Pro yeast Top2
closely resembling their action against prokaryotic enzymes,
supporting a conserved mechanisms of drug action along the
phylogenetic evolution.
CONCLUSIONS
Quinolones can be considered as double-edged drugs:
depending upon fine details in their structure, they can
switch their recognition of the cleavage complex from
bacteria to humans. Of course, this may raise concerns and
hopes. In fact, while cytotoxicity represents a serious
drawback when pursuing antibacterial therapy, it becomes a
desired property when developing anticancer agents.
Although exploitation of topoisomerase II is not new in the
cancer field, the use of novel chemical entities is desirable to
increase the effectiveness of a treatment, in particular by
reducing the onset of drug resistance. Indeed, pleiotropic
mechanisms related to MDR are not likely to be overcome
by antineoplastic quinolones, but these drugs might be useful
in the treatment of cancers with mutated top2s that have
become resistant to the traditionally used enzyme poisons.
The knowledge thus far available on the mechanisms
governing drug selectivity towards eukaryotic vs.
prokaryotic species is as yet far from being fully understood.
It appears that fine tuning processes occur by slightly
changing quinolone structure, that render quinolones more
effective against the bacterial or the mammalian enzyme.
The numerous modifications thus far examined give some
hints of the SAR for selectivity.
For anticancer quinolones, there is no need to preserve
antibacterial efficacy. Hence, chemical functions that are not
suitable to modification in antibacterial chemotherapy can be
proficiently substituted to improve antineoplastic response.
In fact, the SAR referring to antibacterial properties does not
simply hold for antitumor potency. In general, two major
types of modification appear to drive the quinolone action
towards human Top2:
1) Breakage of the zwitterionic properties of the
compounds by modifying either the C-7 basic substituent or
the C-3 carboxyl group, or both will change the electron
density distribution of the quinolone and its protonation
equilibria. It is interesting to note that the keto-carboxyl
O O
F F
O O
N N
O O
H2N
O
QQ58
Fig. (7). Chemical structure of fluoroquinophenoxazine (QQ58) and fluoroquinoanthoxazine (FQA-CR).
Sissi and Palumbo
group is generally preserved in potent antibacterial
quinolones.
2) Increase in the extent of aromatic/condensed rings is
likely to increase the affinity of the quinolone for double
stranded DNA, favoring stacking processes with the planar
structure produced by base pairing. Interestingly, classical
quinolones interact better with single-stranded DNA and are
poorly effective on the B structure of the nucleic acid. They
are, therefore better suited to interfere with plasmids, in
which distortion of base pairing occurs as a result of
supercoiling.
In addition, structural changes that are likely to impair
bactericidal activity do not necessarily grant removal of the
adverse effects known for antibacterial quinolones. Particular
concern can be raised by the reported phototoxic properties
of the classical quinolones, which might represent an
additional drawback in anticancer therapy. However, this
unwanted side effect can be turned into an appreciable
therapeutic tool when considering its potential applications
in photodynamic therapy, as already proposed for
quinobenzoxazines [86].
When introducing structural changes into quinolones,
other cell-affecting mechanisms can be activated. This is
indeed the case for the benzoxazines reported in Fig. (7)
[91]. In fact, the extended planar system introduced with the
aim of further stabilizing stacking interactions with DNA,
renders the new compounds available for efficient binding to
G-quartet structures [92]. This, on turn, raised the possibility
that the enzyme telomerase, activated in the vast majority of
tumor cells, might represent an additional target for
quinolone action and, hence, led to an improved
chemotherapeutic application of the new compounds.
In addition to practical applications, the possibility of
dual targeting for quinolones represents an effective
biochemical tool to dissect the molecular mechanism(s) of
topoisomerase poisoning and to pinpoint similarities and
differences produced by evolution in enzymes having a
unique role in cell survival and proliferation.
In conclusion, quinolones represent a very valuable
family of multi-faceted drugs, the chemical synthesis of,
which is flexible and can be easily adapted to prepare new
congeners with rationally devised structures. This is
witnessed by the description of many thousands of
O O
N N
H2N
FQA-CR
The Quinolone Family Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6 449

derivatives in the literature, and has allowed, and still allows [41] Takei, M.; Fukuda, H.; Yasue, T.; Hosaka, M.; Oomori, Y.
to-day, to accurately describe their QSAR, which is essential Antimicrob. Agents Chemother., 1998, 42, 2678.
[42] Dobson, R.A.; O'Connor, J.R.; Poulin, S.A.; Kundsin, R.B.; Smith,
for effective drug design and optimization, and to discover T.F.; Came, P.E. Antimicrob. Agents Chemother., 1980, 18, 738.
new and unprecedented activities, which makes quinolones [43] Carabateas, P.M.; Brundage, R.P.; Gellote, M.D.; Lorenz, R.R.;
as yet endless source of (pleasant) surprises and, hopefully, Opalka, C.J.; Thielking, W.H., Williams, G.L.; Lesher, G.Y. J.
of further clinically useful drugs. Heterocycl. Chem., 1984, 21, 1875.
[44] Barrett, J.F.; Gootz, T.D.; McGuirk, P.R.; Farrell, C.A.;
Sokolowski, S.A. Antimicrob. Agents Chemother., 1989, 33, 1697.
REFERENCES [45] Peterson, L.R. Clin. Infect. Dis., 2001, 33 Suppl 3, S180.
[46] Robinson, M.J.; Martin, B.A.; Gootz, T.D.; McGuirk, P.R.;
[1] Lesher, G.Y. J. Med. Pharm. Chem., 1962, 5, 1063.
Osheroff, N. Antimicrob. Agents Chemother., 1992, 36, 751.
[2] Kim, O.K.; Ohemeng, K.; Barrett, J.F. Expert Opin. Investig.
[47] Arakawa, H.; Mano, E.; Hakoda, N.; Yoshinari, T.; Nakagawa, S.;
Drugs, 2001, 10, 199.
Okura, A. Anticancer Drug Des., 1996, 11, 221.
[3] Zhanel, G.G.; Ennis, K.; Vercaigne, L.; Walkty, A.; Gin, A.S.;
Embil, J.; Smith, H.; Hoban, D.J. Drugs, 2002, 62, 13. [48] Elsea, S.H.; McGuirk, P.R.; Gootz, T.D.; Moynihan, M.; Osheroff,
N. Antimicrob. Agents Chemother., 1993, 37, 2179.
[4] Heisig, P. Planta Med., 2001, 67, 3.
[49] Yamashita, Y.; Ashizawa, T.; Morimoto, M.; Hosomi, J.; Nakano,
[5] Wang, J.C. Annu. Rev. Biochem., 1985, 54, 665.
H. Cancer Res., 1992, 52, 2818.
[6] Higgins, P.G.; Fluit, A.C.; Schmitz, F.J. Curr. Drug Targets, 2003,
4, 181. [50] Suto, M.J.; Domagala, J.M.; Roland, G.E.; Mailloux, G.B.; Cohen,
[7] Gellert, M.; Mizuuchi, K.; O'Dea, M.H.; Nash, H.A. Proc. Natl. M.A. J. Med. Chem., 1992, 35, 4745.
[51] Alovero, F.L.; Pan, X.S.; Morris, J.E.; Manzo, R.H.; Fisher, L.M.
Acad. Sci. U. S. A., 1976, 73, 3872.
Antimicrob. Agents Chemother., 2000, 44, 320.
[8] Reece, R.J.; Maxwell, A. Crit. Rev. Biochem. Mol. Biol., 1991, 26,
[52] Gootz, T.D.; McGuirk, P.R.; Moynihan, M.S.; Haskell, S.L.
335.
[9] Kato, J.; Nishimura, Y.; Yamada, M.; Suzuki, H.; Hirota, Y. J Antimicrob. Agents Chemother., 1994, 38, 130.
[53] Robinson, M.J.; Martin, B.A.; Gootz, T.D.; McGuirk, P.R.;
Bacteriol, 1988, 170, 3967.
Moynihan, M.; Sutcliffe, J.A.; Osheroff, N. J. Biol. Chem., 1991,
[10] Kato, J.; Nishimura, Y.; Imamura, R.; Niki, H.; Hiraga, S.; Suzuki,
266, 14585.
H. Cell, 1990, 63, 393.
[11] Peng, H.; Marians, K.J. J. Biol. Chem., 1993, 268, 24481. [54] Wentland, M.P.; Lesher, G.Y.; Reuman, M.; Gruett, M.D.; Singh,
B.; Aldous, S.C.; Dorff, P.H.; Rake, J.B.; Coughlin, S.A. J. Med.
[12] Marians, K.J.; Hiasa, H. J. Biol. Chem., 1997, 272, 9401.
[13] Williams, N.L.; Howells, A.J.; Maxwell, A. J. Mol. Biol., 2001, Chem., 1993, 36, 2801.
[55] Fang, K.C.; Chen, Y.L.; Sheu, J.Y.; Wang, T.C.; Tzeng, C.C. J.
306, 969.
[14] Morrison, A.; Cozzarelli, N.R. Cell, 1979, 17, 175. Med. Chem., 2000, 43, 3809.
[15] Kampranis, S.C.; Bates, A.D.; Maxwell, A. Proc. Natl. Acad. Sci. [56] Eissenstat, M.A.; Kuo, G.H.; Weaver, I.J.; Wentland, M.P.;
U. S. A., 1999, 96, 8414. Robinson, R.G.; Klingbeil, K.M.; Danz, D.W.; Corbett, T.H.;
Coughlin, S.A. Bioorg. Med. Chem. Lett., 1995, 5, 1021.
[16] Ullsperger, C., Cozzarelli, N.R. J. Biol. Chem., 1996, 271, 31549.
[57] Kohlbrenner, W.E.; Wideburg, N.; Weigl, D.; Saldivar, A.; Chu,
[17] Levine, C.; Hiasa, H.; Marians, K.J. Biochim. Biophys. Acta., 1998,
1400, 29. D.T. Antimicrob. Agents Chemother., 1992, 36, 81.
[18] Khodursky, A.B.; Peter, B.J.; Schmid, M.B.; DeRisi, J.; Botstein, [58] DoganKoruznjak, J.; Slade, N.; Zamola, B.; Pavelic, K.;
Karminski-Zamola, G. Chem. Pharm. Bull. (Tokyo), 2002, 50, 656.
D.; Brown, P.O.; Cozzarelli, N.R. Proc. Natl. Acad. Sci. U. S. A.,
[59] Tomita, K.; Tsuzuki, Y.; Shibamori, K.; Tashima, M.; Kajikawa,
2000, 97, 9419.
[19] Deibler, R.W.; Rahmati, S.; Zechiedrich, E.L. Genes. Dev., 2001, F.; Sato, Y.; Kashimoto, S.; Chiba, K.; Hino, K. J. Med. Chem.,
2002, 45, 5564.
15, 748.
[20] Willmott, C.J.; Critchlow, S.E.; Eperon, I.C.; Maxwell, A. J. Mol. [60] Wentland, M.P.; Lesher, G.Y.; Reuman, M.; Pilling, G.M.;
Saindane, T.; Perni, R.B.; Eissenstat, M.A.; Weaver, I.J.; Singh, B.;
Biol., 1994, 242, 351.
[21] Wentzell, L.M.; Maxwell, A. J. Mol. Biol., 2000, 304, 779. Rake, J.; Coughlin, S.A. Bioorg. Med. Chem. Lett., 1993, 3, 1711.
[61] Coughlin, S.A.; Danz, D.W.; Robinson, R.G.; Klingbeil, K.M.;
[22] Shea, M.E.; Hiasa, H. J. Biol. Chem., 2000, 275, 14649.
Wentland, M.P.; Corbett, T.H.; Waud, W.R.; Zwelling, L.A.;
[23] Yoshida, H.; Bogaki, M.; Nakamura, M.; Nakamura, S.
Antimicrob. Agents Chemother., 1990, 34, 1271. Altschuler, E.; Bales, E.; Rake, J.B. Biochem. Pharmacol., 1995,
50, 111.
[24] Piddock, L.J. Drugs, 1999, 58 Suppl 2, 11.
[25] Heddle, J.G.; Barnard, F.M.; Wentzell, L.M.; Maxwell, A. [62] Gmunder, H.; Kuratli, K.; Keck, W. Nucleic Acids Res., 1997, 25,
604.
Nucleosides Nucleotides Nucleic Acids, 2000, 19, 1249.
[26] Palu, G.; Valisena, S.; Ciarrocchi, G.; Gatto, B.; Palumbo, M. Proc. [63] Corbett, A.H.; Hong, D.; Osheroff, N. J. Biol. Chem., 1993, 268,
14394.
Natl. Acad. Sci. U. S. A., 1992, 89, 9671.
[27] Sissi, C.; Perdona, E.; Domenici, E.; Feriani, A.; Howells, A.J.; [64] Wentland, M.P.; Aldous, S.C.; Guett, M.D.; Perni, R.B.; Poeles,
R.G.; Danz, D.W.; Klingbeil, K.M.; Peverly, A.D.; Robinson, R.G.;
Maxwell, A.; Palumbo, M. J. Mol. Biol., 2001, 311, 195.
[28] Shen, L.L.; Pernet, A.G. Proc. Natl. Acad. Sci. U. S. A., 1985, 82, Corbett, T.H.; Rake, J.; Coughlin, S.A. Bioorg. Med. Chem. Lett.,
1995, 5, 405.
307.
[65] Chu, D.T.; Hallas, R.; Clement, J.J.; Alder, J.; McDonald, E.;
[29] Critchlow, S.E.; Maxwell, A. Biochemistry, 1996, 35, 7387.
[30] Kampranis, S.C.; Maxwell, A. J. Biol. Chem., 1998, 273, 22606. Plattner, J.J. Drugs Exp. Clin. Res., 1992, 18, 275.
[66] Chu, D.T.; Hallas, R.; Tanaka, S.K.; Alder, J.; Balli, D.; Plattner,
[31] Maxwell, A. Nat. Struct. Biol., 1996, 3, 109.
[32] Berger, J.M.; Gamblin, S.J.; Harrison, S.C.; Wang, J.C. Nature, J.J. Drugs Exp. Clin. Res., 1994, 20, 177.
[67] Clement, J.J.; Burres, N.; Jarvis, K.; Chu, D.T.; Swiniarski, J.;
1996, 379, 225.
Alder, J. Cancer Res., 1995, 55, 830.
[33] Berger, J.M.; Fass, D.; Wang, J.C.; Harrison, S.C. Proc. Natl.
Acad. Sci. U. S. A., 1998, 95, 7876. [68] Kwok, Y.; Sun, D.; Clement, J.J.; Hurley, L.H. Anticancer Drug
Des., 1999, 14, 443.
[34] Chen, G.L.; Yang, L.; Rowe, T.C.; Halligan, B.D.; Tewey, K.M.;
Liu, L.F. J. Biol. Chem., 1984, 259, 13560. [69] Zeng, Q.; Kwok, Y.; Kerwin, S.M.; Mangold, G.; Hurley, L.H. J.
Med. Chem., 1998, 41, 4273.
[35] Woese, C.R. Microbiol. Rev., 1987, 51, 221.
[70] Anderson, V.E.; Gootz, T.D.; Osheroff, N. J. Biol. Chem., 1998,
[36] Forterre, P.; Nadal, M.; Elie, C. Syst. Appl. Microbiol., 1986, 7, 67.
273, 17879.
[37] Sioud, M.; Forterre, P. Biochemistry, 1989, 28, 3638.
[71] Anderson, V.E.; Zaniewski, R.P.; Kaczmarek, F.S.; Gootz, T.D.;
[38] Hussy, P.; Maass, G.; Tummler, B.; Grosse, F.; Schomburg, U.
Osheroff, N. J. Biol. Chem., 1999, 274, 35927.
Antimicrob. Agents Chemother., 1986, 29, 1073.
[72] Elsea, S.H.; Hsiung, Y.; Nitiss, J.L.; Osheroff, N. J. Biol. Chem.,
[39] Hoshino, K.; Sato, K.; Une, T.; Osada, Y. Antimicrob. Agents
Chemother., 1989, 33, 1816. 1995, 270, 1913.
[73] Spitzner, J.R.; Chung, I.K.; Gootz, T.D.; McGuirk, P.R.; Muller,
[40] Akasaka, T.; Kurosaka, S.; Uchida, Y.; Tanaka, M.; Sato, K.;
M.T. Mol. Pharmacol., 1995, 48, 238.
Hayakawa, I. Antimicrob. Agents Chemother., 1998, 42, 1284.
450 Curr. Med. Chem. – Anti-Cancer Agents, 2003, Vol. 3, No. 6 Sissi and Palumbo

[74] Strumberg, D.; Nitiss, J.L.; Dong, J.; Kohn, K.W.; Pommier, Y. J. [83] Chow, K.C.; Macdonald, T.L.; Ross, W.E. Mol. Pharm., 1988, 34,
Biol. Chem., 1999, 274, 28246. 467.
[75] Huff, A.C.; Robinson, R.G.; Evans, A.C.; Selander, K.N.; [84] Fan, J.Y.; Sun, D.; Yu, H.; Kerwin, S.M.; Hurley, L.H. J. Med.
Wentland, M.P.; Rake, J.B.; Coughlin, S.A. Anticancer Drug Des., Chem., 1995, 38, 408.
1995, 10, 251. [85] Yu, H.; Hurley, L.H.; Kerwin, S.M. J. Am. Chem. Soc., 1996, 118,
[76] Strumberg, D.; Nitiss, J.L.; Rose, A.; Nicklaus, M.C.; Pommier, Y. 7040.
J. Biol. Chem., 1999, 274, 7292. [86] Yu, H.; Kwok, Y.; Hurley, L.H.; Kerwin, S.M. Biochemistry, 2000,
[77] Hsiung, Y.; Elsea, S.H.; Osheroff, N.; Nitiss, J.L. J. Biol. Chem., 39, 10236.
1995, 270, 20359. [87] Permana, P.A.; Snapka, R.M.; Shen, L.L.; Chu, D.T.; Clement, J.J.;
[78] Strumberg, D.; Nitiss, J.L.; Dong, J.; Walker, J.; Nicklaus, M.C.; Plattner, J.J. Biochemistry, 1994, 33, 11333.
Kohn, K.W.; Heddle, J.G.; Maxwell, A.; Seeber, S.; Pommier, Y. [88] Kwok, Y.; Zeng, Q.; Hurley, L.H. J. Biol. Chem., 1999, 274,
Antimicrob. Agents. Chemother., 2002, 46, 2735. 17226.
[79] Robinson, M.J.; Osheroff, N. Biochemistry, 1991, 30, 1807. [89] Elsea, S.H.; Westergaard, M.; Burden, D.A.; Lomenick, J.P.;
[80] Corbett, A.H.; Guerry, P.; Pflieger, P.; Osheroff, N. Antimicrob. Osheroff, N. Biochemistry, 1997, 36, 2919.
Agents Chemother., 1993, 37, 2599. [90] Hammonds, T.R.; Foster, S.R.; Maxwell, A. J. Mol. Biol., 2000,
[81] Robinson, M.J.; Corbett, A.H.; Osheroff, N. Biochemistry, 1993, 300, 481.
32, 3638. [91] Kim, M.Y.; Duan, W.; Gleason-Guzman, M.; Hurley, L.H. J. Med.
[82] Osheroff, N.; Corbett, A.H.; Elsea, S.H.; Westergaard, M. Cancer Chem., 2003, 46, 571.
Chemother. Pharmacol., 1994, 34 Suppl, S19. [92] Neidle, S.; Parkinson, G. Nature Rev. Drug Discovery, 2002, 1,
383.