Use of Topoisomerase inhibitors

for treatment of Cancer

By
Shahzada Khurram Syed

Cancer is basically a disease of cells characterized by a
shift in the control mechanisms that govern cell
proliferation and differentiation.
Neoplastic cells proliferate excessively and form local
tumors that can compress or invade adjacent normal
structures.


Introduction
Lymphomas
Originate from lymphatic system
Sarcomas
They originate in the connective tissues like
ligaments, cartilage
Carcinomas
They originate from the epithelial tissues which are
lining of organs like the lungs and liver
Classes of cancer




Leukimias:
is a type of cancer of the blood or bone marrow characterized
by an abnormal increase of immature white blood cells called
"blasts".
Hodgkin: Is a type of lymphoma, which is a cancer originating
from white blood cells called lymphocytes.
When doctors detect the presence of a specific type of
abnormal cell called a Reed-Sternberg cells, the lymphocyte is
called Hodgkin.
Genetic predisposition
exposure to environmental carcinogens.
Chemical carcinogens (particularly those in tobacco
smoke) as well as azo dyes, aflatoxins, asbestos, and
benzene
Radiations
ultra violet radiations
Coal tars



Causes of Cancer
Tumor suppressor genes, may be deleted or damaged,
with resulting neoplastic change.
A single gene in this class, the p53 gene, has been shown to
have mutated from a tumor suppressor gene to an
oncogene in a high percentage of cases of several human
tumors,
Certain herpes and papilloma group DNA viruses and type
C RNA viruses have also been implicated as causative
agents in animal cancers and are responsible for some
human cancers as well.


Causes
The Cell Cycle G0 phase (resting stage): The cell has
not yet started to divide. Cells spend much of their lives
in this phase. Depending on the type of cell, G0 can last
from a few hours to a few years. When the cell gets a
signal to reproduce, it moves into the G1 phase.
G1 phase: During this phase, the cell starts making more
proteins and growing larger, so the new cells will be of
normal size. This phase lasts about 18 to 30 hours
Cell cycle
Cell cycle
S phase: In the S phase, the chromosomes containing the genetic
code (DNA) are copied so that both of the new cells formed will have
matching strands of DNA. The S phase lasts about 18 to 20 hours.

G2 phase: In the G2 phase, the cell checks the DNA and gets ready
to start splitting into 2 cells. This phase lasts from 2 to 10 hours

M phase (mitosis): In this phase, which lasts only 30 to 60 minutes,
the cell actually splits into 2 new cells. This cell cycle is important
because many chemotherapy drugs work only on cells that are
actively reproducing (not cells that are in the resting phase, G0).
Some drugs specifically attack cells in a particular phase of the cell
cycle (the M or S phases


Topoisomerase inhibitors are agents designed to interfere with
the action of topoisomerase enzymes (topoisomerase I and II),
which are enzymes that control the changes in DNA structure by
catalyzing the breaking and rejoining of the phosphodiester
backbone of DNA strands during the normal cell cycle.
In recent years, topoisomerases have become popular targets
for cancer chemotherapy treatments. It is thought that
topoisomerase inhibitors block the ligation step of the cell cycle,
generating single and double stranded breaks that harm the
integrity of the genome. Introduction of these breaks
subsequently leads to apoptosis and cell death.
TOPOISOMERASE INHIBITOR
Topoisomerase inhibitors are often divided according to which
type of enzyme it inhibits.

Topoisomerase I inhibitors: irinotecan, topotecan, camptothecin
and lamellarin D all target type IB topoisomerases,

Topoisomerase II inhibitors: etoposide (VP-16), teniposide,
doxorubicin, daunorubicin, mitoxantrone, amsacrine, ellipticines,
aurintricarboxylic acid, and HU-331, a quinolone synthesized from
cannabidiol.


CLASSIFICATION
These inhibitors are split into two main classes: topoisomerase
poisons, which target the topoisomerase-DNA complex, and
topoisomerase inhibitors, which disrupt catalytic turnover.
Topo II poisons
Examples of topoisomerase poisons include the following:
eukaryotic type II topoisomerase inhibitors (topo II): amsacrine,
etoposide, etoposide phosphate, teniposide and doxorubicin.
These drugs are anti-cancer therapies.
bacterial type II topoisomerase inhibitors (gyrase and topo IV):
fluoroquinolones. These are antibacterials and include such
fluoroquinolones as ciprofloxacin.

TOPOISOMERASE II INHIBITORS
Some of these poisons encourage the forward cleavage reaction
(fluoroquinolones), while other poisons prevent the re-ligation
of DNA (etoposide and teniposide).
Interestingly, poisons of type IIA topoisomerases can target
prokaryotic and eukaryotic enzymes preferentially, making them
attractive drug candidates. Ciprofloxacin targets prokaryotes in
excess of a thousandfold more than it targets eukaryotic topo
IIs. The mechanism for this specificity is unknown, but drug-
resistant mutants cluster in regions around the active site.

The topoisomerases are essential nuclear enzymes for DNA
replication. They control DNA structure by maintaining the
correct super helical state within the cell, as well as by
resolving intertwined DNA strands. This requires the
formation of transient breaks in DNA. Topoisomerase I
generates single-stranded breaks, and topoisomerase II
introduces double-stranded breaks.

All cells require topoisomerases, but fast-growing cancer
cells need more of them. By inhibiting the enzymes, the drugs
selectively inflict more damage on cancer cells than on
normal cells.

Mechanism of actions of
Topoisomerase enzyme
The nuclear enzymes topoisomerase I and II are
critical for DNA function and cell survival, and recent
studies have identified these enzymes as cellular
targets for several clinically active anticancer drugs.

Importance of Topoisomerase
enzymes
Podophyllotoxins ( Etoposide and teniposide )

Mechanism of action:

Block cell cycle: in late S-G2 phase inhibition of
topoisomerase II -- which results in DNA damage
through strand breakage induced by the formation of a
ternary complex of drug, DNA, and enzyme.


Topoisomerase II inhibitors
Topoisomerase inhibitors are cell cycle specific , that is,
they only kill cells that are in a particular phase of cell
division and generally do not have any effect on other
cells. Examples of Topoisomerase inhibitors are
Etoposide and Topotecan

Topoisomerase Inhibitors

Nausea
vomiting
alopecia
significant hematopoietic toxicity
lymphoid toxicity
Adverse Effects of Topoisomerase II
Inhibitors
The drugs are water-insoluble and need to be
formulated in a Cremophor vehicle for clinical use.
These agents are administered via the intravenous route
and are rapidly and widely distributed throughout the
body except for the brain.
Up to 9095% of drug is protein-bound, mainly to
albumin. Dose reduction is required in the setting of
renal dysfunction.
Etoposide has clinical activity in germ cell cancer
Pharmacokinetics of Topoisomerase
II inhibitors
Etoposide has clinical activity in germ cell cancer,
lung cancer, Hodgkin's and non-Hodgkin's
lymphomas, and gastric cancer
High-dose therapy in the transplant setting for breast
cancer and lymphomas.
Teniposide's use is limited to acute lymphoblastic
leukemia
Therapeutic use
Camptothecins
The Camptothecins are natural products that are
derived from the Camptotheca acuminate tree
Mechanism of action
They inhibit the activity of topoisomerase I, the key
enzyme responsible for cutting and relegating single
DNA strands.
Inhibition of the enzyme results in DNA damage.
Topoisomerase I inhibitors
Topotecan is indicated in the treatment of patients of

Advanced ovarian cancer

approved as second-line therapy lung cancer.

Topoisomerase I Inhibitor drugs
The main route of elimination is renal excretion, and
for this reason caution must be exercised in patients
with abnormal renal function, with dosage reduction
being required.

Pharmacokinetics
Primary neutropenia

thrombocytopenia

anemia
Adverse effects
Table. Topoisomerase Inhibitor-Based Cancer Drugs
Product Target Indication Clinical Status Manufacturer
Hycamtin Topoisomerase I Small cell lung cancer Market GlaxoSmithKline
Irinotecan (Campto, Camptosar) Topoisomerase I Colorectal cancer Market Aventis, Pfizer, and Merck
Lucanthone Topoisomerase II Brain tumor Phase II SuperGen
MLN576 Topoisomerase I and II Various tumors Phase I Millennium
Novantrone Topoisomerase II Leukemia and prostate cancer Market OSI
Pegamotecan Topoisomerase I Gastroesopheagal cancer Phase II Enzon
Rubitecan Topoisomerase I Prostate cancer Phase III SuperGen
Topoisomerase II inhibitors (anthracyclines,
epipodophyllotoxins, etc.) are active against several
types of tumours. However, treatment with these
drugs often results in the development of the multi-
drug resistance. Because topoisomerase II-active
drugs have several different modes of action,
different mechanisms of resistance, including
decreased activation and increased detoxification by
glutathione-dependent enzymes, have also been
implicated.
Resistance to Topoisomerase II
inhibitors
Certain clinical limitations of the camptothecin derivatives are.
These include:
1) spontaneous inactivation to a lactone form in blood,
2) rapid reversal of the trapped cleavable complex after drug
removal, requiring prolonged infusions,
3) resistance of cancer cells overexpressing membrane
transporters, and
4) dose-limiting side effects of diarrhea and neutropenia.


Resistance to Topoisomerase I
inhibitors
To circumvent these limitations, Dr. Mark Cushman at Purdue
University and Dr. Yves Pommier at the National Cancer Institute
developed the non-camptothecin family of indenoisoquinoline
inhibitors of Top1. In contrast to the camptothecins, the
indenoisoquinolines are:
1) chemically stable in blood,
2) inhibitors of Top1 cleavable complexes at distinct sites,
3) not substrates of membrane transporters, and
4) more effective as anti-tumor agents in animal models.
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