Antineoplastic Agents



• Introduction – Cell cycle, Cell kill hypothesis

• Drug Resistance to chemotherapy

• General Classification of Antineoplastic agents

• Mechanism of action of alkylating agents

• Uses and toxic / side effects of individual agents

• Antimetabolite drugs

• Mechanism and uses

• Plant products – Podophyllotoxin, Etoposide

• Vinca Alkaloids – Structures, mode of action and uses and
toxic effects

• Miscellaneous compounds – Cisplatin, hydoxy urea,
asparaginase, pipobroman

• Hormones – mode of action and structures, uses and side
effects of individual compounds

• Immunotherapy

• Antibiotics

• Miscellaneous agents


At the end of this
lecture, student will be able to

• Describe various stages of cell cycle

• Explain Drug Resistance to Chemotherapy

• Classify antineoplastic agents

• Discuss the mode of action of Alkylating agents

• Illustrate uses and side effects of Alkylating agents 

• Discuss the different antimetabolites as anticancer drugs

• Explain mechanism of action of antimetabolites

• Discuss the structures, uses of vinca alkaloids

• Describe the mode of action of compounds like cisplatin,

• Explain the mode of action of Hormones for Antineoplastic

• Discuss aspects involved in Immunotherapy

• Discuss the different plant products as anticancer drugs

• Explain mechanism of action of different plant products

• Discuss the different antibiotics as anticancer drugs

• Discuss miscellaneous agents as anticancer drugs


• Antineoplastic agents are the drugs used for the treatment
of cancer

• Cancer or neoplasm refers not to a single disease but to a
group of diseases probably caused by several agents such as certain chemical
compounds, radiant energy, certain viruses and cellular mutation of unknown

• The basic differences between cancer cells and normal
cells are uncontrolled cell proliferation, decreased cellular differentiation,
ability to invade surrounding tissue and ability to establish new growth at
ectopic sites (metastasis)

• Cancer is more difficult to cure than bacterial infections

• There are qualitative differences between human and
bacterial cells, but the differences between normal and neoplastic human cells
are mostly quantitative.

• Immune mechanisms and other host defenses are very
important in killing bacteria and other foreign cells, whereas they play
negligible role in killing cancer cells.

• A single cancer cell can reestablish the tumor, and it is
extremely difficult to effect complete kill of cancer cells

• A higher proportion of resistant cells would be present,
which would mean that retreatment with the same agent would achieve a lesser
response than before

Cell cycle: –

• Cell cycle is divided into ‘s’ phase, where DNA
replication takes place and ‘M’ phase where the mitosis takes place

• Between these two phases is G1 and G2, which are called as
resting phases

• G1 is highly variable because of another phase G0

Typical duration:

• S = 10 – 20 hours

• G2 = 2 – 10 hours

• M = 0.5 – 1 hour

• G1 = highly variable

• Cells in the S or M phase are highly susceptible to
anti-neoplastic agents unlike cells in resting phase which are resistant

• Most anticancer drugs block the biosynthesis or
transcription of nucleic acids or they prevent cell division by interfering
with mitotic spindles.

Cell-kill hypothesis:

• This hypothesis states that the effect of antitumor drugs
on tumor cell populations follow first-order kinetics. This means that the
number of cells killed is proportional to the dose.

• Thus, chemotherapy follows an exponential or log-kill
model in which a constant proportion, not a constant number of cancer cells are

• So, theoretically cancer chemotherapy can never reduce
tumor populations to zero complete eradication requires another effect, such as
the immune response.

Drug resistance

• Drug resistance to chemotherapy usually involves the selection
of certain cell populations

• Populations of drug – resistance cells can be produced by

• Clonal evolution or by Mutation

• Mutagenic agents increase the frequency of generation of drug-resistant

• Many anti-tumor agents are mutagenic.

• Intracellular effects that cause drug resistance may be secondary
to cellular adaptation or altered enzyme levels or properties

• For example, resistance to methotrexate involves increased
levels of the target enzymes, “dihydrofolate reductase”.

Other modes of
resistance to antimetabolites include

• Reduced drug transport into the cells

• Reduced affinity of the molecular target

• Stimulation of alternate biosynthetic pathways 

• Impaired activation or increased metabolism of the drug

• A major factor in resistance to alkylating agents is the
ability of tumor cell to repair DNA lesions such as cross-linkings and breakage
of DNA strands caused by alkylation

• Chemotherapy is not the initial treatment used against

• If the cancer is well defined and accessible, surgery is
preferred skin cancers and certain localized tumors are treated by

• Generally, chemotherapy is important when the tumor is
inoperable or has metastasized.

• Chemotherapy is finding increasing use as an ‘adjuvant’
after surgery to ensure that few cells remain to regenerate the parent tumors
are killed.


Antineoplastic agents are classified as

• Alkylating agents

• Antimetabolites

• Antibiotics

• Plant products

• Hormones

• Miscellaneous agents

• Radioactive isotopes

• Immunotherapy


• Nitrogen mustards

– Mechlorethamine

– Cyclophosphamide

– Melphalan

• Alkyl sulphonate ester

– Busulfan

• Nitrosourea

• Lomustine

• Carmustine

• Aziridine

– Thiotepa


• Folate antagonist

• Methotrexate

• Purine antagonists

– 6-mercapto purine

– 6-thioguanine

• Pyrimidine antagonists

– 5-fluorouracil

– Cytarabine


• Etoposide

• Taxol

• Camphothecin

• Vinblastine

• Vincristine


– Dactinomycin

– Bleomycin

– Mitomycin


– Dromostanolone

– Megestrol


– Cisplatin, Asparginase, Hydroxyurea


– Interferon α2a and 2b

Alkylating agents: (Present working hypothesis)

• Alkylation is defined as the replacement of hydrogen on an
atom by an alkyl group. The alkylation of nucleic acids or proteins involves a
substitution reaction in which a nucleophilic atom (nu) of the biopolymer
displaces a leaving group from the alkylating agent.

Nu-H + alkyl-y → alkyl-nu + H+ + Y

• The reaction rate depends on the nucleophilicity of the
atom (S, N, O), which is greatly enhanced if the nucleophile is ionized.

• Alkylating agents produce cytotoxic, mutagenic and
carcinogenic effects by reacting with cellular DNA.

• They also react with RNA and proteins. The most active
clinical alkylating agents are bi-functional compounds capable of crosslinking
DNA. The cross-linking process can be inter-strand or intra-strand. 

• Inter-strand cross-linking occurs with mechlorethamine or
other ‘two-armed’ mustards.

• Intra-strand cross-linking occurs with busulfan.

• In DNA, ‘7’position (Nitrogen) of guanine – ribose linkage
susceptible to cleavage. This cleavage results in the deletion of guanine and
also cleavage of 8-9 bond in the purine nucleus.

• Other base positions of DNA attacked by alkylating agents
are N-2 and N-3 of guanine; N-3, N-1 & N-7 of adenine; O-6 of thymine; and
N-3 of cytosine.


Use: – in the
treatment of Hodgkin’s disease, lymphomas and mycosis fungoides can be treated
with mechloethamine



• Active orally and parenterally and can be given in
fractionated doses over prolonged periods


• Active against multiple myeloma, chronic lymphocytic
leukemia (CLL) and acute leukemia of children

• In combination with other C.T agents, it cures Burkett’s
lymphoma and acute lymphoblastic leukemia (ALL) in children.

Toxic effects

• Alopecia, nausea and vomiting, leukopenia, sterile
hemorrhagic cystitis. 


• Antimetabolites are compounds that prevent the
biosynthesis or use of normal cellular metabolites

• All the clinical agents are related to metabolites and
cofactors in the biosynthesis of nucleic acids

• They are closely related in structure to the metabolite
that is antagonized

• Most antimetabolites are effective cancer chemotherapeutic
agents via interaction with the biosynthesis of nucleic acids

• Therefore several of the useful drugs used in
antimetabolite therapy are purines, pyrimidines, folates, and related compounds

• Many antimetabolites are enzyme inhibitors, by combining
with the active site as if they were the substrate or cofactor or bind to an
allosteric regulatory site

• Most of these targeted enzymes and processes are involved
in the regulatory steps of cell division and cell/tissue growth

• Sometimes, the antimetabolites must be transformed
biosynthetically into the active inhibitor

• The administration of many purine and pyrimidine
antimetabolites requires the formation of the nucleoside and finally the
corresponding nucleotide for antimetabolite activity

• An antimetabolite and its transformation products may
inhibit several different enzymes involved in tissue growth

• These substances are generally cell cycle specific with
activity in the S phase

• Folate antagonist

• Methotrexate

• Purine antagonists

– 6-mercapto purine

– 6-thioguanine

• Pyrimidine antagonists

– 5-fluorouracil

– Cytarabine

Folic acid



• Methotrexate is the classic antimetabolite of folic acid
structurally derived by N-methylation of the para-aminobenzoic acid residue
(PABA) and replacement of a pteridine hydroxyl by the bioisosteric amino group

• Methotrexate due to its structural similarity with folic
acid inhibits the enzyme dihydrofolate reductase and there by prevent the
formation dihydrofolic acid and tetrahydrofolic acid – which are needed for
nucleic acid synthesis in tumor cell

• This drug competitively inhibits the binding of the
substrate folic acid to the enzyme DHFR, resulting in reductions in the
synthesis of nucleic acid bases, perhaps most importantly, and the conversion
of uridylate to thymidylate as catalyzed by thymidylate synthetase


• In the treatment of acute lymphocytic leukemia and acute
lymphoblastic leukemia.

• Since it has some ability to enter the CNS, it is used in
the treatment and prophylaxis of meningeal leukemia.



• In the treatment of acute lymphocytic leukemia and acute

• Since it has some ability to enter the CNS, it is used in
the treatment and prophylaxis of meningeal leukemia

• 6-mercaptopurine is converted into the corresponding
ribonucleotide, which is potent inhibitor of the conversion of 5phosphoribosyl
pyrophosphate into 5-phosphoribosylamine, which is the rate controlling step in
the synthesis of purines in microbial cells

• Mercaptopurine is used alone or with other chemotherapy drugs
to treat acute lymphocytic leukemia

Toxicity: bone
marrow depression and orogastrointestinal damage


• Thioguanine is converted into its ribonucleotide by the
same enzyme that acts on 6-mercaptopurine. It is converted further into the di-
& triphosphates. These species inhibit most of the same enzymes that are
inhibited by 6-mercaptopurine.

• Thioguanine is also incorporated into RNA and its 2’-deoxy
metabolite’ is incorporated into DNA

• Use:
exclusively for the treatment of leukemias.

Toxicity: bone
marrow depression and orogastrointestinal damage


• In certain tumors uracil is used as the major precursor
for the nucleic acid pyrimidine biosynthesis.

• 5-fluorouracil is the antimetabolite of uracil.

• 5-fluorouracil−−−−−−−−−−→5-fluoro-2-deoxyuridylic
acid−−−−−−−−−−−−−−−−−−−−−−−−−→Thymidylate synthetase (inhibits –powerful
competitive inhibitors)

• (Thymidylate synthetase – responsible for the conversion
of 2deoxyuridylic acid to thymidylic acid – to pyrimidine synthesis in tumor

• The tetrahydrofuranyl derivative of 5-fluorouracil
(TEGAFUR) is active in clinical cancer and less myelosuppressant than

• TEGAFUR is slowly metabolized to 5-fluorouracil and so it
is considered as a pro-drug

• Use: – in the
treatment of carcinomas of breast, colon, pancreas, stomach and rectum

• Topically in the treatment of premalignant keratosis of


• Floxuridine is a fluorinated pyrimidine monophosphate
analogue of 5-fluoro-2′-deoxyuridine-5′-phosphate (FUDR-MP) with antineoplastic

• As an antimetabolite, floxuridine inhibits thymidylate
synthase, resulting in disruption of DNA synthesis and cytotoxicity

• This agent is also metabolized to fluorouracil and other
metabolites that can be incorporated into RNA and inhibit the utilization of
preformed uracil in RNA synthesis

• Floxuridine (FUDR) is a pyrimidine analogue used as an
antineoplastic agent

• Used to treat hepatic metastases of gastrointestinal
adenocarcinomas and for palliation in malignant neoplasms of the liver and
gastrointestinal tract


• Cytarabine is used in the treatment of acute myelogenous
leukemia and CML

• This drug is a deoxycytidine analog

• It is active following intracellular activation to the
nucleotide metabolite ara-CTP.

• The resulting ara-CTP is incorporated into DNA resulting
in chain termination and inhibition of DNA synthesis and function

• Resistance can occur because of decreased activation or
transport and increased catabolic breakdown

• Metabolic breakdown within the GI tract leads to poor

• Toxicities include myelosuppression, leukopenia and
thrombocytopenia, nausea and vomiting anorexia, diarrhea, and mucositis


• Azathioprine is in a class of medications called immunosuppressants

• Azathioprine for the treatment of autoimmune disorders is
significantly associated with an increased risk for developing acute myeloid
leukemia (AML)




• Bleomycin is a glycopeptide antibiotic complex isolated
from Streptomyces verticillus



• Daunorubicin is used to treat leukemia and other cancers.
It belongs to a class of drugs known as anthracyclines and works by slowing or
stopping the growth of cancer cells.


• Doxorubicin is a type of chemotherapy drug called an

It slows or stops the growth of cancer cells by blocking an
enzyme called topo isomerase II

• Acute lymphoblastic leukemia (ALL)

• Acute myeloblastic leukemia (AML)

• Bone sarcoma

• Breast cancer

Plant products

• Podophyllotoxin, obtained from the resin of the May apple,
podophyllum peltatum has antineoplastic activity but is highly toxic.

• This lignin inhibits mitosis by destroying the structural
organization of the mitotic apparatus. The newer analogues of podophyllotoxin
eg., Etoposide are much better antineoplastic agents. This drug will inhibit
topoisomerase II unlike podophyllotoxin



• In combination with other chemotherapeutic agents

• It is the first choice treatment for small cell lung

• In refractory testicular tumors, acute non-lymphocytic
leukemia, Hodgkin’s disease, non-Hodgkin’s lymphomas and Kaposi’s sarcoma.

Toxicity: –bone
marrow suppression, alopecia, nausea and vomiting.


• Are a family of important antitumor agents from plants.

• Isolated from periwinkle ‘Catharanthus rosea’.

• Composed of indole – containing moiety, Vindoline.

• Four closely related compounds have anti-tumor activity:
Vincristine, Vinblastine, Vinrosidine, Vinleurosine.

• Among this Vincristine and Vinblastine are proved clinical

Mode of Action:

• Vinca alkaloids cause mitotic arrest by promoting the
dissolution of microtubules in cells.

• Microtubule crystals containing the alkaloids are formed
in the cytoplasm.

• Vinblastine is the most active compound whereas
Vincristine is the only compound to cause irreversible inhibition of mitosis.


Effective against acute leukemia

In the treatment of Hodgkin’s disease (combination

Other tumors that respond to vincristine in combination with
other chemotherapeutic agents include lympho-sarcoma, reticulum cell sarcoma,
rhabdomyosarcoma, neuroblastoma and Wilms’ tumor.

Toxicity: –

Neurological (loss of deep tendon reflexes, pain and muscle


• Used for the palliative of a variety of neoplastic

• One of the most effective single agents against Hodgkin’s

• In the treatment of advanced testicular germinal cell

• In lymphocytic lymphoma, histiocytic lymphoma, mycosis fungoides,
Koposi’s sarcoma, Leterer-Siwe disease, resistant choriocarcinoma and carcinoma
of breast


• Leukopenia

• Gastrointestinal and neurological symptoms

• Cellulitis and phlebitis.




Miscellaneous compounds


Mode of action: –

• It interacts with DNA and Intra-strand crosslinks are
produced and causes changes in DNA conformation that affect replication.

Use: –

• Used (in combination with bleomycin and vinblastin) for
metastatic testicular tumors

• For the remission of metastatic ovarian tumors (a single
agent or in combination)

• Other uses: in penile cancer, bladder cancer, cervical
cancer, head & neck cancer and small cell cancer of the lung.

Toxicity: –

• Cumulative renal insufficiency associated with renal
tubular damage

• Myelo suppression, nausea & vomiting and ototoxicity


• Enzyme isolated commercially from E. Coli and E.

• Asparagine is required for the biosynthesis of proteins.

• Although normal cells can synthesise ‘asparagine’ tumors
such as acute lymphoblastic leukemia (ALL) lack this ability and depend on
exogenous compound.

• Administration of Asparaginase reduces the concentration
of Asparagine in plasma, making it unavailable to the leukemia cells.

Use: –

• To induce remissions in ALL and non-Hodgkin’s lymphoma.

Side effects: –

• Hypersensitivity, gastrointestinal damage, hepatic
toxicity and pancreatitis.



• Against hematological cancers, especially polycythemia
vera and chronic granulocytic leukemia.


• Steroids hormones including estrogen, androgens, progestin
and glucocorticoids act on the appropriate target tissues at the level of transcription.

• Target cells contain, in their cytoplasm specific protein
receptors with very high affinities for the hormones.

• Binding of the hormone to the receptor transform the
receptor structure, followed by migration of the resulting complex into the nucleus.

• In the nucleus, the complex interacts with an acceptor
site to influence transcription.

• Normal and well – differentiated neoplastic target cells
have a no. of hormone receptors and they depend on the hormones for

• Less-differentiated neoplastic cells become independent of
hormonal control and lose their specific receptors.

• Thus, some neoplasms are hormone dependent and responsive
to hormone-based therapy, whereas others are independent and unresponsive

Miscellaneous Compounds

• Cisplatin, cisplatinum, or cis-diamminedichloroplatinum
(II), is a well-known chemotherapeutic drug. It has been used for treatment of
numerous human cancers including bladder, head and neck, lung, ovarian, and
testicular cancers


• Mitotane is used to treat cancer of the adrenal gland that
cannot be treated with surgery

• It works by slowing growth or reducing the size of the


• Cells of neoplastic potential are continually produced in
the human body and our immune surveillance system destroys them.

• The development of tumors implies that this system is not functioning

• Evidence for this factors in carcinogenesis includes

A high rate of cancer in organ
– transplant patients whose immune systems are suppressed by drugs such as

A high correlation between
cancer and immune-deficiency diseases such as bacterial and viral infections.

Stimulation of the body’s
immune system is a valuable method of cancer treatment because it can eradicate
the neoplastic cells completely.

• Interferons are secreted by cells in response to viral
infections or other chemical or biological inducers.

• Three major classes of interferons – α, β, γ were

• They bind to specific high-affinity receptors on cell
surfaces, which induces a sequence of intracellular events, including the
indication of enzymes.

• This process causes enhancement of natural killer cell
activity, inhibition of certain oncogenes and increased specific cytotoxicity of
lymphocytes for target cells.

• Interferons alfa – 2a, alfa – 2b and alfa – n3 promote the
immunological response to neoplastic cells which results in significant

• These are the drugs of choice for treatment of hairy cell

• Also effective against cell cancer, multiple myeloma,
melanoma, kaposi’s sarcoma (clinical trials)

Alfa – 2a:

• Highly purified protein containing 165 amino acids.

Used: – in the
treatment of hairly cell leukemia and chronic myelogenous leukemia

Side effects:
hypersensitivity, (contraindicated), Flu-like syndrome, consisting of fever,
fatigue, myalgia, headache and chills, gastrointestinal and CNS symptoms.

Alfa – 2b:

• It is a highly purified protein

Use: – Used for
the treatment of Hairy cell leukemia.

• In treating melanoma and renal cell carcinoma.

Side effects: –
flu-like syndrome, CNS effects and cardiovascular effects including
hypotension, arrhythmia or tachycardia