.

Pyrimidine analogues
Pyrimidine analogues is nucleoside analog antimetabolites which mimic the structure of metabolic
pyrimidines.

Examples:
5-Fluorouracil (5FU) which inhibits thymidylate synthase.
Floxuridine (FUDR)
Cytarabine (Cytosine arabinoside)
6-azauracil (6-AU)

Pyrimidine

Fluorouracil

Floxuridine

Gemcitabine
Cytarabine

6-Azauracil

Fluorouracil
A pyrimidine analog that is an antineoplastic antimetabolite. It interferes with DNA
synthesis by blocking the thymidylate synthetase conversion of deoxyuridylic acid to
thymidylic acid.

This compound belongs to the class of organic compounds known as halopyrimidines.

These are aromatic compounds containing a halogen atom linked to a pyrimidine ring.
Pyrimidine is a 6-membered ring consisting of four carbon atoms and two nitrogen
centers at the 1- and 3- ring positions.

Indication:

For the topical treatment of multiple actinic or solar keratoses. In the 5% strength it is
also useful in the treatment of superficial basal cell carcinomas when conventional
methods are impractical, such as with multiple lesions or difficult treatment sites.
Fluorouracil injection is indicated in the palliative management of some types of cancer,
including colon, esophageal, gastric, rectum, breast, biliary tract, stomach, head and neck,
cervical, pancreas, renal cell, and carcinoid.

Mechanism of action:

The precise mechanism of action has not been fully determined, but the main mechanism
of fluorouracil is thought to be the binding of the deoxyribonucleotide of the drug
(FdUMP) and the folate cofactor, N510-methylenetetrahydrofolate, to thymidylate
synthase (TS) to form a covalently bound ternary complex. This results in the inhibition
of the formation of thymidylate from uracil, which leads to the inhibition of DNA and
RNA synthesis and cell death. Fluorouracil can also be incorporated into RNA in place of
uridine triphosphate (UTP), producing a fraudulent RNA and interfering with RNA
processing and protein synthesis.
 Floxuridine
An antineoplastic antimetabolite that is metabolized to fluorouracil when administered
by rapid injection. Floxuridine is available as a sterile, nonpyrogenic, lyophilized powder
for reconstitution. When administered by slow, continuous, intra-arterial infusion, it is
converted to floxuridine monophosphate. It has been used to treat hepatic metastases of
gastrointestinal adenocarcinomas and for palliation in malignant neoplasms of the liver
and gastrointestinal tract.

Indication:

For palliative management of gastrointestinal adenocarcinoma metastatic to the liver,

when given by continuous regional intra-arterial infusion in carefully selected patients
who are considered incurable by surgery or other means. Also for the palliative
management of liver cancer (usually administered by hepatic intra-arterial infusion).

Mechanism of action:

Floxuridine is rapidly catabolized to 5-fluorouracil, which is the active form of the drug.
The primary effect is interference with DNA synthesis and to a lesser extent, inhibition of
RNA formation through the drug's incorporation into RNA, thus leading to the
production of fraudulent RNA. Fluorouracil also inhibits uracil riboside phophorylase,
which prevents the utilization of preformed uracil in RNA synthesis. As well, the
monophosphate of floxuridine, 5-fluoro-2'-deoxyuridine-5'-phosphate (FUDR-MP)
inhibits the enzyme thymidylate synthetase. This leads to the inhibition of methylation
of deoxyuridylic acid to thymidylic acid, thus interfering with DNA synthesis.

Cytarabine
A pyrimidine nucleoside analog that is used mainly in the treatment of leukemia,
especially acute non-lymphoblastic leukemia. Cytarabine is an antimetabolite
antineoplastic agent that inhibits the synthesis of DNA. Its actions are specific for the S
phase of the cell cycle. It also has antiviral and immunosuppressant properties

Indication:

For the treatment of acute non-lymphocytic leukemia, acute lymphocytic leukemia and
blast phase of chronic myelocytic leukemia.

Mechanism of action:

Cytarabine acts through direct DNA damage and incorporation into DNA. Cytarabine is
cytotoxic to a wide variety of proliferating mammalian cells in culture. It exhibits cell phase
specificity, primarily killing cells undergoing DNA synthesis (S-phase) and under certain
conditions blocking the progression of cells from the G1 phase to the S-phase. Although
the mechanism of action is not completely understood, it appears that cytarabine acts
through the inhibition of DNA polymerase. A limited, but significant, incorporation of
cytarabine into both DNA and RNA has also been reported.
 6-azauracil
An analog of inosine with a methylthio group replacing the hydroxyl group in the 6-position

This compound belongs to the class of organic compounds known as glycosylamines.

These are compounds consisting of an amine with a beta-N-glycosidic bond to a
carbohydrate, thus forming a cyclic hemiaminal ether bond (alpha-amino ether).

Purine analogues
Purine analogues are antimetabolites that mimic the structure of metabolic purines.

Azathioprine is the main immunosuppressive cytotoxic substance. It is widely used in

transplantations to control rejection reactions. It is nonenzymatically cleaved to 6-
mercaptopurine that acts as a purine analogue and an inhibitor of DNA synthesis. By
preventing the clonal expansion of lymphocytes in the induction phase of the immune
response, it affects both the cell and the humoral immunity. It also successfully
suppresses autoimmunity.
Mercaptopurine
Thiopurines such as thioguanine are used to treat acute leukemias and remissions in
acute granulocytic leukemias
Fludarabine inhibits function of multiple DNA polymerases, DNA primase, and DNA
ligase I, and is S phase-specific (since these enzymes are highly active during DNA
replication).
Pentostatin and cladribine are adenosine analogs that are used primarily to treat hairy
cell leukemia.

Purine

Mercaptopurine

Thioguanine


Fludarabine

Azathioprine

Pentostatin

Azathioprine (AZA):
is an immunosuppressive drug used in organ transplantation and autoimmune diseases and
belongs to the chemical class of purine analogues. Synthesized originally as a cancer drug and a
prodrug for mercaptopurine in 1957, it has been widely used as an immunosuppressant for more
than 50 years.

Azathioprine acts as a prodrug for mercaptopurine, inhibiting an enzyme required for the
synthesis of DNA. Thus, it most strongly affects proliferating cells, such as the T cells and B cells
of the immune system.

The main adverse effect of azathioprine is bone marrow suppression, which can be life-
threatening, especially in people with a genetic deficiency of the enzyme thiopurine S-
methyltransferase. It is also listed by the International Agency for Research on Cancer as a group
1 carcinogen (carcinogenic to humans).

Azathioprine is produced by a number of manufacturers under different brand names (Azasan,

Imuran ).

Azathioprine is a prodrug, a substance that is not an active drug itself but is activated in the body.
This happens in several steps; at first it is slowly and almost completely converted to 6-
mercaptopurine (6-MP) by reductive cleavage of the thioether (S). This is mediated by
glutathione and similar compounds in the intestinal wall, the liver and on red blood cells, without
the aid of enzymes. 6-MP is metabolized analogously to natural purines, giving thioguanosine
triphosphate (TGTP) and thio-deoxyguanosine triphosphate (TdGTP) via thioinosine
monophosphate (TIMP) and several further intermediates. On a second path, the sulfur atom of
6-MP and TIMP is methylated. The end products of azathioprine metabolism are thiouric acid
(38%) and various methylated and hydroxylated purines, which are excreted via the urine.

Mechanism of action:
The purine molecule is the framework for two of the four bases that occur in DNA, adenine and
guanine. Consequently, blocking the synthesis of purine also hinders DNA synthesis and thus
inhibits the proliferation of cells, especially fast-growing cells without a method of nucleotide
salvage ("recycling"), such as lymphocytes. Two types of lymphocytes, T cells and B cells, are
particularly affected by the inhibition of purine synthesis.

Azathioprine's active metabolite methyl-thioinosine monophosphate (MeTIMP) is a purine

synthesis inhibitor that works by blocking the enzyme amidophosphoribosyltransferase, possibly
among others. Additional mechanisms, mediated by other metabolites, have been discovered
several decades after the drug's introduction, mainly in the 1990s and 2000s: Thioguanosine
triphosphate (TGTP) is incorporated into RNA, compromising its functionality. It also interacts
with the GTP-binding protein Rac1, blocking upregulation of the protein Bcl-xL and thus sending
activated T cells into apoptosis (a kind of programmed cell death). The closely related thio-
deoxyguanosine triphosphate (TdGTP) is built into DNA. Thioinosinic acid impedes later steps of
DNA synthesis via enzymes such as adenylosuccinate synthase and IMP dehydrogenase.
Moreover, azathioprine blocks the downstream effects of CD28 costimulation, a process required
for T cell activation. In vivo data indicate inflammatory bowel disease patients treated with
azathioprine have more mononuclear cells that have undergone apoptosis than untreated
controls, indicating this mechanism may be responsible for the in vivo response to the drug in
this disease.

Pharmacokinetics

Azathioprine is absorbed from the gut to about 88%. Bioavailability varies greatly between
individual patients, between 30 and 90%, because the drug is partly inactivated in the liver.
Highest blood plasma concentrations, counting not only the drug itself but also its metabolites,
are reached after one to two hours; and the average plasma half-life is 26 to 80 minutes for
azathioprine and three to five hours for drug plus metabolites. 20 to 30% are bound to plasma
proteins while circulating in the bloodstream.

Azathioprine is a prodrug, a substance that is not an active drug itself but is activated in the body.
This happens in several steps; at first it is slowly and almost completely converted to 6-
mercaptopurine (6-MP) by reductive cleavage of the thioether (S). This is mediated by
glutathione and similar compounds in the intestinal wall, the liver and on red blood cells, without
the aid of enzymes. 6-MP is metabolized analogously to natural purines, giving thioguanosine
triphosphate (TGTP) and thio-deoxyguanosine triphosphate (TdGTP) via thioinosine
monophosphate (TIMP) and several further intermediates. On a second path, the sulfur atom of
6-MP and TIMP is methylated. The end products of azathioprine metabolism are thiouric acid
(38%) and various methylated and hydroxylated purines, which are excreted via the urine

Mechanism of action

The purine molecule is the framework for two of the four bases that occur in DNA, adenine and
guanine. Consequently, blocking the synthesis of purine also hinders DNA synthesis and thus
inhibits the proliferation of cells, especially fast-growing cells without a method of nucleotide
salvage ("recycling"), such as lymphocytes. Two types of lymphocytes, T cells and B cells, are
particularly affected by the inhibition of purine synthesis.

Azathioprine's active metabolite methyl-thioinosine monophosphate (MeTIMP) is a purine

synthesis inhibitor that works by blocking the enzyme amidophosphoribosyltransferase, possibly
among others. Additional mechanisms, mediated by other metabolites, have been discovered
several decades after the drug's introduction, mainly in the 1990s and 2000s: Thioguanosine
triphosphate (TGTP) is incorporated into RNA, compromising its functionality. It also interacts
with the GTP-binding protein Rac1, blocking upregulation of the protein Bcl-xL and thus sending
activated T cells into apoptosis (a kind of programmed cell death). The closely related thio-
deoxyguanosine triphosphate (TdGTP) is built into DNA. Thioinosinic acid impedes later steps of
DNA synthesis via enzymes such as adenylosuccinate synthase and IMP dehydrogenase.
Moreover, azathioprine blocks the downstream effects of CD28 costimulation, a process required
for T cell activation. In vivo data indicate inflammatory bowel disease patients treated with
azathioprine have more mononuclear cells that have undergone apoptosis than untreated
controls, indicating this mechanism may be responsible for the in vivo response to the drug in
this disease.

Mercaptopurine
Mercaptopurine (also called 6-mercaptopurine, 6-MP or its brand name Purinethol) is an
immunosuppressive drug. It is a thiopurine.,

Medical uses

It is used to treat leukemia, non-Hodgkin's lymphoma in children, polycythemia vera, psoriatic

arthritis, and inflammatory bowel disease (such as Crohn's disease and ulcerative colitis). It has
demonstrated some in vitro effectiveness against Mycobacterium paratuberculosis.

Mechanisms of action

Mercaptopurine (6-MP) competes with the purine derivatives hypoxanthine and guanine for
the enzyme HGPRT and is itself converted to thio inosine monophosphate (TIMP).
o TIMP inhibits several chemical reactions involving inosinic acid (IMP), including the
conversion of IMP to xanthylic acid (XMP) and the conversion of IMP to adenylic
acid (AMP) via adenylosuccinate (SAMP).
o In addition, 6-methylthioinosinate (MTIMP) is formed by the methylation of TIMP.
Both TIMP and MTIMP have been reported to inhibit glutamine-5-
phosphoribosylpyrophosphate amidotransferase, the first enzyme unique
to the de novo pathway for purine ribonucleotide synthesis. Experiments
indicate that radiolabeled mercaptopurine may be recovered from the DNA
in the form of deoxythioguanosine.
Some mercaptopurine is converted to nucleotide derivatives of 6-thioguanine (6-TG) by the
sequential actions of inosinate (IMP) dehydrogenase and xanthylate (XMP) aminase,
converting TIMP to thioguanylic acid (TGMP).
Animal tumors that are resistant to mercaptopurine often have lost the ability to convert
mercaptopurine to TIMP. However, it is clear that resistance to mercaptopurine may be
acquired by other means as well, particularly in human leukemias.
It is not known exactly which of any one or more of the biochemical effects of
mercaptopurine and its metabolites are directly or predominantly responsible for cell
death.

6-MP ribonucleotide inhibits purine nucleotide synthesis and metabolism by inhibiting an

enzyme called Phosphoribosyl pyrophosphate amidotransferase (PRPP Amidotransferase). PRPP
Amidotransferase is the rate limiting enzyme of purine synthesis. This alters the synthesis and
function of RNA and DNA. Mercaptopurine interferes with nucleotide interconversion and
glycoprotein synthesis.

Pharmacogenetics
The enzyme thiopurine S-methyltransferase (TPMT) is responsible, in part, for the inactivation of
6-mercaptopurine. TPMT catalyzes the methylation of 6-mercaptopurine into the inactive
metabolite 6-methylmercaptopurine - this methylation prevents mercaptopurine from further
conversion into active, cytotoxic thioguanine nucleotide (TGN) metabolites. Certain genetic
variations within the TPMT gene can lead to decreased or absent TPMT enzyme activity, and
individuals who are homozygous or heterozygous for these types of genetic variations may have
increased levels of TGN metabolites and an increased risk of severe bone marrow suppression
(myelosuppression) when receiving mercaptopurine.[12] In many ethnicities, TPMT
polymorphisms that result in decreased or absent TPMT activity occur with a frequency of
approximately 5%, meaning that about 0.25% of patients are homozygous for these variants.
However, an assay of TPMT activity in red blood cells or a TPMT genetic test can identify patients
with reduced TPMT activity, allowing for the adjustment of mercaptopurine dose or avoidance of
the drug entirely. The FDA-approved drug label for mercaptopurine recommends testing for
TPMT activity to identify patients at risk for myelotoxicity. Indeed, testing for TPMT activity is
currently one of the few examples of pharmacogenetics being translated into routine clinical care

Tioguanine
An antineoplastic compound which also has antimetabolite action. The drug is used in the
therapy of acute leukemia.

Tioguanine (INN, BAN), or thioguanine (AAN, USAN), commonly referred to as 6-thioguanine

(6-TG) is a medication used to treat a number of types of leukemia.[1] It has been used less
frequently in recent years because of safety concerns. However, it is becoming more widely used
for treating ulcerative colitis and some autoimmune diseases.

It belongs to the thiopurine family of drugs that also include mercaptopurine and azathioprine,
which are examples of antimetabolites. It is a purine analogue of the nucleobase guanine. It is a
pale yellow, odorless, crystalline powder.

Indication:

Acute leukemias in both adults and children

Chronic myelogenous leukemia
Inflammatory bowel disease, especially ulcerative colitis
Psoriasis.

Mechanism of Action:

6-Thioguanine is a thio analogue of the naturally occurring purine base guanine. 6-thioguanine
utilises the enzyme hypoxanthine-guanine phosphoribosyltransferase (HGPRTase) to be
converted to 6-thioguanine monophosphate (TGMP). High concentrations of TGMP may
accumulate intracellularly and hamper the synthesis of guanine nucleotides via the enzyme
Inosine monophosphate dehydrogenase (IMP dehydrogenase).

TGMP is converted by phosphorylation to thioguanine diphosphate (TGDP) and thioguanine

triphosphate (TGTP). Simultaneously deoxyribosyl analogs are formed, via the enzyme
ribonucleotide reductase. The TGMP, TGDP and TGTP are collectively named 6-thioguanine
nucleotides (6-TGN). 6-TGN are cytotoxic to cells by: (1) incorporation into DNA during the
synthesis phase (S-phase) of the cell; and (2) through inhibition of the GTP-binding protein (G
protein) Rac1, which regulates the Rac/Vav pathway. An additional effect may be derived from
the incorporation of 6-thioguanine into RNA. This yields a modified RNA strand which cannot be
read by the ribosomes.

Fludarabine
Indication:

For the treatment of adult patients with B-cell chronic lymphocytic leukemia (CLL) who have not
responded to or whose disease has progressed during treatment with at least one standard
alkylating-agent containing regimen.

Mechanism of action:

Fludarabine phosphate is rapidly dephosphorylated to 2-fluoro-ara-A and then phosphorylated

intracellularly by deoxycytidine kinase to the active triphosphate, 2-fluoro-ara-ATP. This
metabolite appears to act by inhibiting DNA polymerase alpha, ribonucleotide reductase and
DNA primase, thus inhibiting DNA synthesis. The mechanism of action of this antimetabolite is
not completely characterized and may be multi-faceted.

Pentostatin
This compound belongs to the class of organic compounds known as imidazodiazepines. These
are organic compounds containing an imidazole ring fused to a diazepine ring. Imidazole is 5-
membered ring consisting of three carbon atoms, and two nitrogen centers at the 1- and 3-
positions. Diazepine is a 7-membered ring consisting of five carbon and two nitrogen atoms.

Indication:

For the treatment of hairy cell leukaemia refractory to alpha interferon.

Mechanism of action:

Pentostatin is a potent transition state inhibitor of adenosine deaminase (ADA), the greatest
activity of which is found in cells of the lymphoid system. T-cells have higher ADA activity than B-
cells, and T-cell malignancies have higher activity than B-cell malignancies. The cytotoxicity that
results from prevention of catabolism of adenosine or deoxyadenosine is thought to be due to
elevated intracellular levels of dATP, which can block DNA synthesis through inhibition of
ribonucleotide reductase. Intracellular activation results in incorporation into DNA as a false
purine base. An additional cytotoxic effect is related to its incorporation into RNA. Cytotoxicity is
cell cycle phase-specific (S-phase).
References:

1-Drug bank web site supported by the Canadian Institutes of Health Research

2- Wikipedia, the free encyclopedia.