Antiviral therapies

BIO 2032
July 2013
(a) Bullet shaped rabies. (b) The segmented helical virus of
influenza. (c) A bacteriophage with an icosahedral head and
helical tail. (d) An enveloped icosahedral herpes simplex
virus. (e) The unenveloped polio virus. (f) The icosahedral
human immunodeficiency virus with spikes on its envelope.
The infection
and
replication
depends on
whether a
virus is
enveloped or
non-
enveloped,
has DNA or
RNA genome
and also on
the cell type.
 Stages
• The first step in the replication process
is attachment.
• During attachment the virus adsorbs to a
susceptible host cell.
• High specificity exists between virus and cell,
and the envelope receptor.
 Stages
• The next step is penetration of the virus or
the viral genome into the cell.
• May occur by endocytosys; or the membrane
fusion.
• The virus envelop fuses with the cell
membrane.
• The replication steps of the process occur
next.
• The protein capsid is stripped away from the
genome, and the genome is freed in the cell
cytoplasm.
• If the genome is positive RNA, the genome
may act as a messenger RNA molecule.
• It is translated for the synthesis of enzymes
and other proteins.
• The enzymes are used for the synthesis of viral
genomes and capsomeres and the assembly of
these components into new viruses.
• If the genome is negative sense mRNA, it is
used to transcribe postive RNA.
• The latter may be used as mRNA to make
proteins.
Antiviral therapies target viral cycle
• The drugs used to treat viral infections target
attachment, uncoating, replication, integration or
any of the steps involved in viral particle
production.
• #1) These drugs interfere with a virus-specific
function.
• The virus function may be unique to the virus.
• The virus function e.g. DNA replication may be
similar to the host function but may be much
less susceptible to the drug.
• #2)Drugs may interfere with a cellular function
so that the virus cannot replicate.
• To be specific, the anti-viral drug must only kill
virus-infected cells.
• This could be done so that drugs get activated
only in cells infected by the virus.
 Antiviral therapies
• Unlike bacteria viruses are not really living organisms.
• Antibiotics are ineffective against virus infections.
• To control infections by viruses, antiviral chemotherapy
and vaccines have been the mainstay.
• Of the two approaches, vaccines are preferred because
• 1) They are comparatively cheaper to produce.
• 2) They are safe.
• 3) They offer long-lasting immunity.
 Antiviral theprapies
• Even the use of vaccines has not been
universally successful.
• Vaccines for viruses such as influenza,
retroviruses and herpesviruses, have been
hard to develop.
• 1) Some change their antigens rapidly.
• 2) Others have long latent periods after
infection such that one may not realize they
are infected.
 Antiviral chemotherapy
• There are examples of antiviral drugs that
have been successful in controlling infections.
• These include RANTES, acyclovir and AZT.
• They are not broad spectrum as the case is for
antibiotics.
• Some serve as a peptide that mimics the
receptor such as soluble CD4 protein.
• This would bind HIV gp120 and stop it binding
to the receptor on the cell surface.
• They may bind to the CCR5 chemokine
receptor and interfere with virus attachment.
• They may down-regulate the co-receptor by
endocytosis making it more difficult for the
virus to bind.
• They however, cannot completely stop viral
replication.
• Some viral particles always seems to be able
to circumvent the drug-induced blockade.
 Examples
• #1) Natural products of plants. These may be
single or mixtures of several plant chemicals.
• They may inhibit virus replication or interfere
with virus-host cell interaction.
• This blocks virus infection and spread.
• #2) Synthetic oligonucleotides,
oligosaccharides, simple inorganic and organic
compounds and
• #3) Nucleoside analogues.
• Some interfere with virus interaction with
receptors.
• For example HIV interacts with CD4 receptors.
• Drugs are available that interfere with this
interaction process.
• Others have been developed to interfere with
virus uncoating.
• Others interfere with virus replication or
transcription.
Inhibitors of herpes DNA polymerase
• The DNA polymerase of the herpes simplex
has been shown to be sensitive to drugs that
target this enzyme.
• Acyclovir inhibits the function of herpes DNA
polymerase.
• There are several factors that have been taken
into consideration for such drugs.
• (1) The drug must be taken up only into
infected cells.
• (2) The actual inhibitory molecule should be
generated inside the infected cell by
enzymatic activity
• (3) The inhibitor should have a selective effect
on a virus enzyme.
• Acyclovir demonstrates all of the above
characteristics.
 Inhibitors of viral reverse
transcriptase
• AZT and the majority of other compounds act
as chain terminators.
• AZT triphosphate binds to and inhibit virus RT
more effectively than normal cellular DNA
polymerases and so some antiviral specificity
is achieved.
• It is toxic.
 Nucleoside analogues
• These have the structure of nucleosides.
• Many occur in nature and are synthesized and
tested in the laboratory.
• Examples are those used against herpesvirus
as inhibitors.
Acyclovir
• When animal host cells get infected by a virus,
they produce antibodies.
• These are protein molecules cells use to
inhibit and prevent an infection.
• They neutralize viral infectivity.
• Vaccines are substances developed to boost
antibody production in the body.
• A virus coat protein when it interacts with a
receptor causes cells to make antibodies.
• This is called an antigen (= antibody generator).
• There is usually a specific part of a virus coat
protein that interacts with cells to cause antibody
production.
• This part is called an epitope.
• Antibodies that inhibit viral infectivity usually
reacts with just a few epitopes on one or two
surface antigens of the infecting agent.
 Endogenous proteins
• Endogenous antigens are those produced
within cells of the body.
• They include:
• A). Viral proteins produced during viral
replication,
• B). Proteins produced by intracellular bacteria
such as Rickettsias and Chlamydias during
their replication.
 Endogenous proteins
• C). Proteins that have escaped into the cytosol
from the phagosome of phagocytes such as
antigen-presenting cells.
• D). Tumor antigens produced by cancer cells.
• E) Self peptides from human cell proteins.
 Body reaction
• When a cell gets infected the body marks it for
destruction.
• This is done by placing peptide epitopes from
these endogenous antigens on their surface
by way of MHC-I molecules.
• Cytotoxic T-lymphocytes (CTLs) are then able
to recognize peptide/MHC-I complexes.
• They use T-cell receptors (TCRs) and CD8
molecules and kill the cells to which they bind.
• 1. Endogenous antigens, such as viral proteins,
pass through proteasomes.
• Proteosomes are cell machinery that degrade
proteins into a series of peptides.
• 2. The peptides are transported into the rough
endoplasmic reticulum (ER) by a transporter
protein called TAP.
3. The peptides then bind to the grooves of
newly synthesized MHC-I molecules.
• 4. The endoplasmic reticulum transports the
MHC-I molecules with bound peptides to the
Golgi complex.
• 5. The Golgi complex, in turn, transports the
MHC-I/peptide complexes by way of an
exocytic vesicle to the cytoplasmic membrane.
• They become anchored to the membrane.
• Here, the peptide and MHC-I/peptide
complexes can be recognized by CTLs by way
of TCRs.
• CTLs and CD8 molecules have shapes
complementary to the MHC-1/peptide
complex.
• A vaccine therefore becomes ineffective if the
protective epitopes are subject to pronounced
antigenic drift.
• This is the case seen in influenza.
• Infected cells which express viral antigen on
their cell surface may be killed.
• This process is facilitated by antibodies
produced in response to viral infection.
• Antibody may facilitate the removal of debris
(a scavenging mechanism)
 An effective vaccine
• It must activate Antigen-Presenting Cells to
initiate antigen processing and producing
interleukins.
• It must cctivation of both T and B cells to give
a high a high yield of memory cells.
• Generation of Th and Tc cells to several
epitopes, to overcome the variation in the
immune response in the population due to
MHC variations.
 Examples of vaccines
• Live whole virus vaccines
• Killed whole virus vaccines
• Subunit vaccines;- purified or recombinant
viral antigen
• Recombinant virus vaccines
• DNA vaccines
 Live Vaccines
• These are made in several ways including:
• #1) Use of a related virus from another
animal.
• Example was the use of cowpox to prevent
smallpox.
• #2) Administration of pathogenic or partially
attenuated virus by an unnatural route.
• This reduces the virulence of the virus.
• Production of temperature sensitive mutants.
 Inactivated whole virus vaccines
• The virus is treated with heat or a chemical.
• The outer virion coat should be left intact but
the replicative function should be destroyed.
• The chemicals used include formaldehyde or
beta- propiolactone.
• The traditional agent for inactivation of the
virus is formalin
Potential problems with live vaccines
• Underattenuation
• Mutation leading to reversion to virulence
• Preparation instability
• Contaminating viruses in cultured cells
• Heat lability
• Should not be given to immunocompromized
or pregnant patients
 Problems
• Killed vaccines
• Incomplete inactivation
• Increased risk of allergic reactions due to large
amounts of antigen involved
 Subunit Vaccines
• These are made through the purification of
virus and viral antigens.
• You identify and the peptide sites of the virus
coat protein that are highly antigenic.
• You purify these and use them as vaccines.
antigenic sites of viral antigens, from which
highly purified subunit vaccines can be
produced
• Example of purified subunit vaccines is the HA
vaccines for influenza A and B.