‫معهد واكادميية النهال‬ DR.

Marwan Imad

Antineoplastic Agents
1-Alkylating Agents
The alkylating agents are a class of drugs that are capable of forming covalent bonds with
important biomolecules. The major targets of drug action are nucleophilic groups present
on DNA (especially the 7-position of guanine); however, proteins and RNA among others
may also be alkylated.
Alkylation of DNA is thought to lead to cell death.

Potential mechanisms of cell death include activation of apoptosis caused by p53 activation
and disruption of the template function of DNA.
Most of the currently used alkylating agents are nonselective regarding the sequence of DNA with
which they react. Therefore, it is uncertain whether alkylation will lead to a cytotoxic event.

Resistant to alkylating agents can result from:

1) Decreased cellular uptake
2) Increased inactivation by detoxifying nucleophilic thiols such as glutathione
3) Increased DNA repair processes
4) Decreased drug activation when this necessary for generation of an alkylating species

NITROGEN MUSTARDS
Mustards such as mechlorethamine are classified as dialkylating agents in that one
mustard molecule can alkylate two nucleophiles.
The initial acid–base reaction is necessary to release the lone pair of electrons on nitrogen,
which subsequently displaces chloride to give the highly reactive aziridinium cation.
Nucleophilic attack can then occur at the aziridinium carbon to relieve the small ring strain
and neutralize the charge on nitrogen.
 ‫معهد واكادميية النهال‬ DR.Marwan Imad
Mechlorethamine is highly reactive, in fact, too reactive and therefore nonselective, making it
unsuitable for oral administration and necessitating direct injection into the tumor. In cases of
extravasation (drug escapes from the tumor into the underlying tissue), the antidote sodium
thiosulfate (Na2S2O3), a strong nucleophile, may be administered.
The lack of selectivity of mechlorethamine led to attempts to improve on the agent. One
rationale was to reduce the reactivity by reducing the nucleophilicity of nitrogen, thereby
slowing aziridinium cation formation. This could be accomplished by replacement of the
weakly electron-donating methyl group with groups that were electron withdrawing. This is
seen in the case of chlorambucil and melphalan by attachment of nitrogen to a phenyl ring.
Reactivity was reduced such that these compounds could be administered orally.

THIOTEPA
Thiotepa containing the thiophosphoramide functionality was found to be more stable than
the oxa-analog (TEPA). Thiotepa incorporates a less reactive aziridine ring compared with
that formed in mechlorethamine. The adjacent thiophosphoryl is electron withdrawing and,
therefore, reduces the reactivity of the aziridine ring system.
Monoalkylation is also possible as a result of aziridine formation via hydrolysis of thiotepa.
 ‫معهد واكادميية النهال‬ DR.Marwan Imad
ORGANOPLATINUM COMPOUNDS
There are several organometallic compounds based on platinum that play a central role
in many cancer treatment protocols. the dichloro species is maintained in the blood stream
as a result of the relatively high chloride concentration. Movement into the tumor cells is
accomplished by passive diffusion or carrier-mediated transport. Once inside the tumor cell, the
drug encounters a lower chloride concentration and one chloro group is substituted by a water
molecule in a process known as aquation. This serves to “trap” the molecule in the cell as a
result of ionization. Reaction with DNA occurs preferentially at the N-7 of guanine of two
adjacent guanine residues resulting in primarily (95%) intrastrand cross -links.
Cisplatin administration is also associated with significant nephrotoxicity and neurotoxicity that is
dose limiting. These factors lead to the development of less reactive platinum compounds such as
carboplatin and oxaliplatin in which the leaving group was incorporated into a chelate.

NITROSOUREAS
General structural requirement:
1) Urea moiety
2) Nitroso group linked to the urea
3) Electronegative atom with good leaving properties

These compounds are reasonably stable at pH = 4.5 but undergo both acid and base
catalyzed decomposition at lower and higher pH, respectively.
alkylation of DNA involves abstraction of the NH proton, which is relatively acidic (pK a =
8–9), followed by rearrangement to give an isocyanate and a diazohydroxide.
The diazohydroxide, upon protonation followed by loss of water, yields a diazo species that
decomposes to a reactive carbocation. The isocyanate functions to carbamylate proteins and RNA,
whereas the carbocation is believed to be the agent responsible for DNA alkylation. Alternative
mechanisms of decomposition have also been proposed involving formation of chlorovinyl
carbocations. In those cases where there is a chloroethyl moiety attached to the N-nitroso urea
functionality, crosslinking of DNA occurs. Alkylation occurs preferentially at the N-7 position of
guanine with minor amounts of alkylation at guanine O-6.
 ‫معهد واكادميية النهال‬ DR.Marwan Imad
Detoxification pathways of the nitrosoureas are also possible and can play a role in
resistance to this group of agents. Two major routes of inactivation have been
identified.The first of these involves dechlorination, which is facilitated by CYP participation
and involves cyclization to give 4,5-dihydro-[1,2,3]oxadiazole and the isocyanate, which is
still capable of carbamylating proteins. The second route involves denitrosation.

PROCARBAZINE

2-ANTIMETABOLITES

Antimetabolites are compounds closely related in structure to a cellular precursor molecule. These
antimetabolites are similar enough in structure in many cases to interact with the normal cellular
process but differ in a manner sufficient to alter the outcome of that pathway.
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.
The antimetabolite drugs may exert their effects by several individual mechanisms
involving enzyme inhibition at active, allosteric, or related sites.
Often the administered drug is actually a prodrug form of an antimetabolite and requires
activation in vivo to yield 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.
The purine and pyrimidine antimetabolites are often compounds incorporated into nucleic acids and
the nucleic acid polymers (DNA, RNA, etc.). The antifolates are compounds designed to interact at
cofactor sites for enzymes involved in the biosynthesis of nucleic acid bases.

Pyrimidine Drugs
The pyrimidine derivative 5-fluorouracil (5-FU) was designed to block the conversion of
uridine to thymidine.
The normal biosynthesis of thymidine involves methylation of the 5-position of the
pyrimidine ring of uridine.

5-Fluorouracil is activated by conversion to the corresponding nucleotide species, 5-fluoro-2-

deoxyuridylic acid. The resulting 5-fluoro-2"-deoxyuridylic acid is a powerful inhibitor of
thymidylate synthetase, the enzyme that converts 2"-deoxyuridylic acid to thymidylic acid.
 ‫معهد واكادميية النهال‬ DR.Marwan Imad

Mechanism of action of 5-FU is by replacement of the hydrogen at the 5-position of uracil

with a fluorine results in antimetabolite drug, leading to the formation of a stable covalent
complex composed of 5-FU, thymidylate synthase and cofactor.

Attempts at chemical modification of 5-FU to protect from catabolic events have produced several
prodrug forms, which are converted via in vivo metabolic and/or chemical transformation to the
parent drug 5-FU foe example capecitabine. The activation sequence of it include:
1) Carbamate hydrolysis
2) Deamination
3) Hydrolysis of sugar moiety to yield 5-FU
 ‫معهد واكادميية النهال‬ DR.Marwan Imad
CYTARABINE

Cytarabine is used in the treatment of acute myelogenous leukemia. This drug is a

deoxycytidine analog. It is active following intracellular activation to the nucleotide
metabolite. The resulting metabolite 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 bioavailability, so it given intravenously.

FLOXURIDINE

The drug is used to treat metastatic GI adenocarcinoma. The mechanism of action of this
fluoropyrimidine deoxynucleoside analog involves metabolic conversion to 5-fluorouracil
(5-FU) metabolites resulting in inhibition of TS thus disrupting DNA synthesis, function,
and repair. Resistance can occur because of increased expression of TS.

Purine Drugs

The design of antimetabolites based on purine structure began with isosteric

thiol/sulfhydryl group to replace the 6-hydroxyl group of hypoxanthine and guanine.
This purine requires bioactivation to its ribonucleotide, 6-thioinosinate (6-MPMP), by the
enzyme HGPRT.

6-MERCAPTOPURINE
 ‫معهد واكادميية النهال‬ DR.Marwan Imad
The mechanism of action includes incorporation of mercaptopurine into DNA and RNA
via the triphosphate metabolite. This incorporation inhibits synthesis and function of the
resulting modified DNA or RNA. The parent drug is inactive and requires phosphorylation
for activity. Resistance can occur via decreased expression of the activating enzymes or
increased expression of the major catabolic enzymes.

THIOGUANINE
The drug is available in 40-mg tablets for oral use. Thioguanine is used to treat acute
nonlymphocytic leukemia. The mechanism of action involves incorporation of the
triphosphate into DNA and RNA, resulting in inhibition of processing and function.
Intracellular phosphorylation is required for activity and inhibition of purine biosynthesis.
Resistance can include decreased expression of the activating enzyme, decreased drug
transport, and/or increased expression of catabolic enzymes. The oral absorption of
thioguanine is poor, and the drug does not appear to cross the blood-brain barrier. Major
metabolic pathways involve deamination or methylation.

Antifolates

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.
The conversion of -OH to -NH2 increases the basicity of N-3 and yields greater enzyme
affinity. 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, the conversion of uridylate to thymidylate as catalyzed by thymidylate synthetase.
 ‫معهد واكادميية النهال‬ DR.Marwan Imad
3-ANTIBIOTICS AND NATURAL PRODUCTS

are derived from natural sources with several of these being obtained from microbial
sources (antibiotics). Many of the antineoplastic antibiotics are produced by the soil fungus
Streptomyces. Both the antibiotic and natural product classes have multiple inhibitory
effects on cell growth; however, they primarily act to disrupt DNA function and cell division.
There are several mechanisms by which these agents target DNA, including intercalation,
alkylation, and strand breakage either directly or as a result of enzyme inhibition.

Intercalation is a process by which a planar molecule of the appropriate size inserts

itself between adjacent base pairs of DNA and in so doing, it causes a local unwinding
that may disrupt the normal template function of DNA.
The overall result of these interactions is to cause a local bend or kink in DNA resulting in a local
shape distortion. This may produce several effects but is often associated with inhibition of normal
DNA function and inhibition of topoisomerase enzymes. Topoisomerase enzymes are responsible
for the unwinding and relaxation of DNA so that transcription may occur.

Actinomycins

The actinomycins are a group of compounds that are isolated from various species
of Streptomyces, all of which contain the same phenoxazone chromophore but
differ in the attached peptide portion.
From this group emerged actinomycin D, which is known as dactinomycin.

Dactinomycin binds noncovalently to double-stranded DNA by partial intercalation

between adjacent guaninecytosine bases resulting in inhibition of DNA function.
the planar phenoxazone ring, which facilitates intercalation between DNA base pairs. The
peptide loops are located within the minor groove and provide for additional interactions.

Mechanism of action
1) The primary effect of this interaction is the inhibition of DNA-directed RNA synthesis and
specifically RNA polymerase. 2) DNA synthesis may also be inhibited, and the agent is
considered cell cycle specific for the G1 and S phases. 3) Inhibition of topoismerase II also
occurs such that the enzyme-DNA complex is stabilized and strand breakage may be seen.
 ‫معهد واكادميية النهال‬ DR.Marwan Imad
Resistance is caused by a decreased ability of tumor cells to take up the drug
and P-glycoprotein (Pgp)-mediated efflux.

Anthracyclines

are characterized by a planar oxidized anthracene nucleus fused to a cyclohexane ring

that is subsequently connected via a glycosidic linkage to an amino sugar.

Doxorubicin

the accepted mechanism involves intercalation followed by inhibition of

topoisomerase II resulting in strand breakage leading to apoptosis.
Resistance involves decreased expression of topoisomerase II and mutations in this enzyme that
decrease binding of the drug. The agent may be actively secreted from cancer cells.

Epipodophyllotoxins

are semisynthetic derivatives of podophyllotoxin.

Etoposide acts on topoisomerase II stabilizing the cleavable complex leading to single-
and double-strand breaks. If enough breaks are initiated, apoptosis is activated. Etoposide
is believed to bind to topoisomerase II in the absence of DNA, because it shows little
tendency to interact with DNA alone. The etoposide-topoisomerase II complex then binds
DNA, and strand cleavage occurs.
 ‫معهد واكادميية النهال‬ DR.Marwan Imad
There are several mechanisms by which cells become resistant to the epipodophyllotoxins
including increased efflux by Pgp as seen for many of the other natural products. Additionally,
topoisomerase II levels may decrease or develop altered binding sites with lower affinity for these
agents. Increased DNA repair mechanisms may also decrease the effectiveness of these
agents.There are mechanisms by which double-strand breaks in DNA can be repaired.

Vinca Alkaloids
The alkaloids are composed of a catharanthine moiety containing the indole subunit and
the vindoline moiety containing the dihydroindole subunit joined by a carbon–carbon bond.
Vincristine and vinblastine differ only in the group attached to the dihydroindole nitrogen,
which is a methyl group in vinblastine and a formyl group in vincristine.

Vinca alkaloids bind to tubulin disrupting formation and function of the mitotic spindle.The
mitotic spindle is composed of the microtubules, which function as part of the cell’s
cytoskeleton and are important in maintaining cellular shape.

MITOMYCIN C
Mitomycin is sometimes included as an alkylating agent but is included here because it
is a naturally occurring material.
The drug contains what would appear to be reactive functionalities, including the quinone
and aziridine functionalities, both or which would be thought to be susceptible to
nucleophilic attack; however, the reactivity of these functionalities is reduced because of
steric and electronic effects in the parent molecule. It was reasoned that selective
activation could be achieved in a reductive environment such as that found in an area of
low oxygen content. This is known to occur in tumors where the fast-growing cells often
grow beyond the blood supply that would normally provide oxygen.
Mitomycin C is capable of being activated and alkylating DNA in an anaerobic
environment but there is actually little selectivity for hypoxic cells. Activation can occur
enzymatically by both one- and twoelectron processes. Reductive enzymes such as
NADPHCYP reductase and DT-diaphorase have been implicated in these processes.