17.

Antiviral agents

Structure
Life cycle (herpes simplex)
 Drugs against viruses
Virus inside cell - hidden from immune defence and drugs...

Use host cell biochemistry - few unique targets...

...few drugs on the market!

1. HIV
2. Herpes
3. Hepatitis Targets should be:
4. Influenza
1. Important to virus life cycle
2. Differ from human proteins
3. Common to different viruses
4. Important in early stages of infection
 Drugs against DNA-viruses
Aciclovir
 Drugs against DNA-viruses
Aciclovir
 Drugs against DNA-viruses
Cidofovir against viruses lacking thymidine kinase
 Drugs against RNA-viruses
Structure and life cycle of HIV
Drugs against RNA-viruses

Three classes of drugs:

1. Nucleoside reverse transcriptase inhibitors

2. Non-nucleoside reverse transcriptase inhibitors
3. Protease inhibitors
Reverse transcriptase inhibitors
Non-nucleoside reverse transcriptase
inhibitors
HIV protease inhibitors
HIV protease inhibitors — TS analogs
HIV protease inhibitors — TS analogs
HIV protease inhibitors — TS analogs
HIV protease inhibitors — TS analogs
HIV protease inhibitors — TS analog
Saquinavir
 NEWS
Drugs against RNA-viruses — flu p911 Eggs-tra
evidence: Scientists
p914 Under fire:
NIH scientists say
p916 Rebel with a
cause: Transgendered
set high burden they feel punished researcher Ben Barres
of proof for fertility for the misdeeds of fights for women in
finding. a few colleagues. science.

Threat of pandemic brings flu drug back to life

© 2005 Nature Publishing Group http://www.nature.com/naturemedicine

With an effective avian influenza vaccine
nowhere on the horizon, health officials are
scrambling for ways to head off a pandemic.
Governments have been purchasing millions of
doses of Roche’s Tamiflu, but would that alone
be enough to head off a pandemic? Should other
drugs be stockpiled? Are other drugs available?
The influenza drug Relenza, available since
1999, had been a footnote to Tamiflu but, after
glowing reports in August of its relative effective-
ness, has jumped back into the spotlight.
Relenza, marketed by GlaxoSmithKline, and
Roche’s Tamiflu both bind to the active site of the
virus’ neuraminidase protein, rendering the virus
unable to escape its host cell and infect others.
In August Germany announced it would buy
1.7 million doses of Relenza as part of its bird
flu preparedness strategy. “Germany’s purchase
shows that countries are starting to take a
balanced view of influenza preparedness,” says
Simon Tucker, head of research at Melbourne-
based Biota, which developed Relenza.
Bulk orders of Tamiflu from the UK, France
and other countries to cover 20% or more
of their population are pushing Roche’s
production capacity to the limit. But Relenza,
which was the first neuraminidase inhibitor
on the market, claims only one percent of the
growing flu drug market. The drug has suffered
from lackadaisical marketing efforts, according
to a lawsuit that Biota, which gets a percentage of
sales, is bringing against GlaxoSmithKline.
Relenza has also been unpopular in part
because it must be breathed in with an inhaler.
GlaxoSmithKline is considering repackaging
Relenza as a shot or nebulizer to increase sales.
Relenza is at least as effective as Tamiflu and
has fewer side effects, including nausea and
headaches, according to an article published
13 August (Lancet 366, 533–534; 2005). The
report, based on data compiled from the
companies’ clinical trials and from subsequent
studies, also says there is no evidence of
resistance to Relenza, compared with resistance
levels of up to 18% in those taking Tamiflu
(Lancet 364, 759–765; 2004). The researchers
recommend stockpiling both.
The Tamiflu resistance data are based on stud-
ies in Japan, which in past years has consumed
nearly 75% of the drug’s worldwide supply.
Other influenza drugs, such as amantadine, have
already been rendered ineffective against bird flu
by misuse (Nature 435, 1009; 2005).
“In a pandemic situation, resistance is a major
concern,” says University of Wisconsin virologist
Yoshihiro Kawaoka, who led the study in Japan.
He adds that Relenza has not been used widely
enough to judge its potential for creating resis-
tance. “It has been underestimated,” he says.
Meanwhile, Biota is conducting phase 1 trials,
boosted by a $5.5 million National Institutes of
Health grant, of a neuraminidase inhibitor that
could be taken only once a week—compared
with twice a day for Relenza and Tamiflu.
But will any—or a combination—of the
drugs help? A pandemic virus would require
quick access to the drugs. After the first few
days of infection, says Graeme Laver, whose
work led to the design of neuraminidase
inhibitors, “you may be able to stop the
virus, but the immune response would likely
kill you.” To prevent misuse of the drugs, he
adds, they should be sold in tandem with an
influenza diagnostic test.
David Cyranoski, Tokyo

Russia

Canada

Netherlands

Virus First
Kazakhstan Korea strain reported
Mongolia Cambodia H5N1 Jan 2004
Canada H7N3 Feb 2004
China Japan
China H5N1 Nov 1996
(mainland)
Hong Kong H5N1 May 1997
Indonesia H5N1 Jan 2004
Laos Kazakhstan H5N1 July 2005
Vietnam Korea H5N1 Dec 2003
Thailand
Mongolia H5N1 Aug 2005
Malaysia
Netherlands H7N7 Feb 2003
Russia

Kimberly Caesar
H5N1 July 2005
Cambodia
Thailand H5N1 Jan 2004
Indonesia Vietnam H5N1 Jan 2004

Room to roam: W ith u n pre dictable migratory ro utes, th e H5N1 virus—which has cause d th e d eath or d estructio n of more than 150 millio n birds in
so uth east Asia—is spreading toward Euro p e. Kazak hstan, Russia, M o ngolia an d Tib et are th e latest to register cases.
* As of 2 0 August 2 0 0 5.

NATURE MEDICINE VOLUME 11 | NUMBER 9 | SEPTEMBER 2005 909

 Drugs against RNA-viruses
Structure and life cycle of influenza virus
Neuraminidase inhibitors
 Neuraminidase inhibitors

C2 “flattened” in TS!
 Neuraminidase inhibitors

TS analogs with sp2-hybridized C2:

 Neuraminidase inhibitors

Added ionic interactions:

 Neuraminidase inhibitors

Carbocyclic TS-analogs:
 Neuraminidase inhibitors

Carbocyclic TS-analogs:
 Neuraminidase inhibitors

Oseltamivir (Tamiflu):
 Neuraminidase inhibitors
Tamiphosphor of September 26 2007:

Published on Web 09/12/2007

Synthesis of Tamiflu and its Phosphonate Congeners Possessing Potent

Anti-Influenza Activity
Jiun-Jie Shie,† Jim-Min Fang,*,†,‡ Shi-Yun Wang,† Keng-Chang Tsai,† Yih-Shyun E. Cheng,†
An-Suei Yang,† Shih-Chia Hsiao,† Ching-Yao Su,† and Chi-Huey Wong†,§
The Genomics Research Center, Academia Sinica, Taipei, 11529, Taiwan. ‡Department of Chemistry,
National Taiwan UniVersity, Taipei 106, Taiwan. §Department of Chemistry and the Skaggs Institute of
Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037

Received June 1, 2007; E-mail: jmfang@ntu.edu.tw

Influenza remains a major health problem for humans and

animals.1 At present, four drugs are approved for influenza
prophylaxis and treatment:2 amantadine and rimantadine act as the
M2 ion channel blockers, whereas Tamiflu (the phosphate salt of
oseltamivir ethyl ester) and Relenza (zanamivir) inhibit the activity
of neuraminidase (NA). The NA inhibitors (NAIs) are designed to
have (oxa)cyclohexene scaffolds to mimic the oxonium transition-
state in the enzymatic cleavage of sialic acid.3 Tamiflu (1, shown
in Scheme 1) is an orally administrated anti-influenza drug.4 On
hydrolysis by hepatic esterases, the active carboxylate, oseltamivir
(2, also known as GS4071), is exposed to interact with three Figure 1. Molecular models of oseltamivir 2 (left panel) and the
arginine residues (Arg118, Arg292, and Arg371) in the active site phosphonate compound 3a (right panel) in the active site of influenza virus
neuraminidase (N1 subtype). The complex of the phosphonate compound
of NA.3 3a has more extensive hydrogen bonding interactions (8 pairs ligand-NA
The phosphonate group is generally used as a bioisostere of H-bonds) with key residues in the NA active site than the oseltamivir-NA
carboxylate in drug design.5 In comparison with the carboxylate- complex (6 pairs ligand-NA H-bonds)
guanidinium ion pair, a phosphonate ion exhibits stronger electro-
Table 1. Inhibitory Activities against Wild-Type and Mutant
static interactions with the guanidinium ion. Our preliminary Influenza Virus Neuraminidases
molecular docking experiments (Figure 1) using the known N1
neuraminidase inhibition, IC50 (nM)
crystal structure (PDB code: 2HU4)3c reveal that the putative
phosphonate inhibitor 3a indeed binds strongly with the triarginine compd Wt (WSN)a Mut (WSN)b Wt (Hanoi)c Mut (Hanoi)d
residues of NA, in addition to other interactions exerted by the 2 5.90 (( 0.62) 295 (( 31) 62.9 (( 5.7) 971 (( 54)
C3-pentyloxy, C4-acetamido, and C5-amino groups in the binding 3e 0.30 (( 0.05) 526 (( 44) 13.3 (( 1.0) 1210 (( 490)
pocket similar to the NA-oseltamivir complex. Because the 13a 4.10 (( 0.51) 252 (( 31) 160 (( 32) 1150 (( 380)
13be 0.12 (( 0.02) 7.39 (( 0.67) 1.82 (( 0.11) 19.5 (( 1.4)
previously reported methods4 for the synthesis of oseltamivir/ 14a 36700 NDf NDf NDf
Tamiflu are not amenable to exchange of the C-1 carboxyl group 14be 3200 NDf NDf NDf
to a phosphonate group, we thus explored a novel approach to the
a NA from influenza virus A/WSN/1933 (H1N1). b NA (H274Y) from
synthesis of both oseltamivir/Tamiflu and the phosphonate conge-
influenza virus A/WSN/1933 (H1N1). c NA from influenza virus A/Hanoi/
ners using D-xylose as an appropriate chiral precursor (Scheme 1). 30408/2005 (H5N1). d NA (H274Y) from influenza virus A/Hanoi/30408/
In brief, our present synthetic method is straightforward to 2005 (H5N1). e As the ammonium salt depicted in Scheme 1. f Not
culminate in an enantioselective synthesis of Tamiflu, oseltamivir, determined.
 Quick questions:
1. What is the difference between phase I and phase II clinical trials?

2. Which part of the molecule below would you expect to be the most labile in
the gastrointestinal tract?
HO
O

HO O

3. Describe one way to stabilize it.

4. A group of atoms in a molecule can be replaced with a different group of

atoms that mimics them. What is the replacing group of atoms called?

1. Pharmacophore X. Isostere 2. Lead structure

 Neuraminidase inhibitors
Tamiphosphor of September 26 2007:
Published on Web 09/12/2007

O
u and its PhosphonateO Congeners Possessing
O
O
Potent O
O P
Anti-Influenza
O Activity O P
O
O
OEt
2xNH4
Fang,*,†,‡ Shi-Yun Wang,† Keng-Chang Tsai,† Yih-Shyun E. Cheng,† AcHN
† Shih-Chia Hsiao, † Ching-Yao Su,† andAcHN
AcHN 2xNH4 †,§
Chi-Huey Wong HN NH
NH3 NH2
Center, Academia Sinica, Taipei, 11529, Taiwan. ‡Department of Chemistry,
H2PO4 NH2
ty, Taipei 106, Taiwan. §Department of Chemistry and the Skaggs Institute of
s Research Institute, 10550 Tamiphosphor
North Torrey Pines Road,
Tamiflu La Jolla, CaliforniaActive
92037 against NAI resistant mutants
EC50 31 nM EC50 4.7 nM EC50 19 nM
Received June 1, 2007; E-mail: jmfang@ntu.edu.tw

blem for humans and

pproved for influenza
rimantadine act as the
(the phosphate salt of
ivir) inhibit the activity
(NAIs) are designed to
he oxonium transition-
id.3 Tamiflu (1, shown
nti-influenza drug.4 On
arboxylate, oseltamivir
to interact with three Figure 1. Molecular models of oseltamivir 2 (left panel) and the
g371) in the active site phosphonate compound 3a (right panel) in the active site of influenza virus
neuraminidase (N1 subtype). The complex of the phosphonate compound
 Search

q
Related Story
Keeping One Step
April 4, 2005 Ahead Of The Flu
Volume 83, Number 14 [C&EN, Feb. 28, 2005]
pp. 47-52
Business

August 29, 2005

PREPARING FOR AVIAN FLU E-mail this article
to a friend
Volume 83, Number 35
pp. 20-22
Much controversy surrounds U.S. plans for defeating a Print this article
possible influenza pandemic E-mail the editor Roche Raises Output Of Bird Flu Drug
BETTE HILEMAN, C&EN WASHINGTON But firm will not meet demand for its antiviral drug
Tamiflu if an avian flu pandemic breaks out
The rapid growth of a
particularly nasty strain of on (–)-shikimic
Jean-Françoisacid: Tremblay,a Diels-Alder-based
C&EN Hong Kong one that uses furan and ethyl acrylate as starting
avian influenza has world materials andin another
These days, Hong Kong, it'sthat relies
still possible onintocatalytic
to walk hydrogenation
a drugstore, hand over HK$200 (U.S. of an isophthalic acid derivative
health officials very worried. followed by
$25) to enzymatic
a clerk, and walk out withdesymmetrization.
a box of 10 capsules of Tamiflu,And Frost
the antiviral drug has
that showsbegun to develop a microbial
September 12, Little can be done to halt the promise in treating humans for the deadly avian flu. Local pharmacists say they face no
2005 disease's spread, and unless
synthesis ofdifficulty
particular aminoshikimic acid, which
in stocking Tamiflu (oseltamivir phosphate) could reduce
and that public demandthefor it isneed for azide chemistry if used as a
not particularly strong. F. Hoffmann-La Roche, which makes the drug, says 10 capsules are
Vol. 83, Iss. 37 nations do more to prepare starting material
enough (Org.
to treat a bout Lett. 2004, 6, 1585). So far, however, none of these alternatives has
of the flu.
View Current Issue measured upwilltobecome
the harder
commercial
for a possible pandemic, the But the drug to find if the flu route
strain nowin terms
spreading of bird
in Asian cost and efficiency.
populations
death toll from this outbreak mutates into a form that can spread rapidly among people and precipitates a bird flu pandemic.
Back Issues could be very large. In the spring of 2003, when Hong Kong was tensely fighting off SARS (severe acute
respiratory syndrome), a teenager posted on a website the fake news that Hong Kong had
2005 Go! been declared a diseased harbor, something that would disrupt food shipments to the territory.
Already this avian influenza This caused thousands of people to rush to supermarkets in the middle of the afternoon to
stock up on essentials like staple foods and bottled water.
is endemic in poultry and
wild birds in 10 Asian Roche is well aware of the likelihood that demand for Tamiflu would surge during a bird flu
countries, and it has killed at COLLECTIVE ACTION A farmer herds his pandemic, not just in Hong Kong, but worldwide. The company insists there is no shortage at
present. But for the past few years, it has been advising health administrations around the
least 49 people. Most ducks in Thai Binh province, Vietnam. Ducks world to stock up on this neuraminidase inhibitor. “We have been very clear that if a pandemic
here often carry the avian influenza virus.
infectious disease experts STR/AFP/GETTY IMAGES
were to start tomorrow, the demand would be so steep that we could not guarantee that a
government could get its supply,” David Reddy, head of Roche's pandemic task force, says.
believe that a worldwide
influenza pandemic is almost inevitable within the next few years. The Few governments have been heeding Roche's advice. The company says it takes about 12
months to produce Tamiflu, which is one of the company's more complex drugs. At present,
United Nations World Health Organization has urged all member states the company requires 12–18 months to supply new orders. A few countries, including Norway,
to make pandemic preparedness plans and begin implementing them. Ireland, the U.K., and New Zealand, have enough capsules to treat up to 40% of their
An expert group convened by WHO last November concluded, "The populations. Fewer than 20 countries, however, have stocked up on Tamiflu, many have not
placed orders, and the supply of the drug worldwide is enough to treat only about 40 million
unpredictability of influenza viruses and the speed with which people.
transmissibility can improve means that the time for preparedness
To help overcome this apparent lack of preparedness, Roche says it will donate 30 million
planning is now." capsules of Tamiflu-enough for 3 million treatments-to the World Health Organization. The
capsules would allow WHO to attempt to contain a pandemic in its early stages, regardless of
Last August, the U.S. government unveiled a draft pandemic Chemical & Engineering News
whether the stricken area has its own stock of the drug.
preparedness plan. But some public health experts consider it highly ISSN 0009-2347
flawed. They claim that it puts way too much emphasis on vaccines, Copyright © 2005
Join ACS which take at least six months to produce and can be stockpiled for
only about 18 months, and too little emphasis on antiviral medicines
that can be stored for five years. They say that once a pandemic starts,
probably in Asia, it is likely to reach the U.S. in less than three months,
long before large supplies of vaccine are available.

The U.S. plan lays out no concrete strategies for allocating limited
supplies of vaccines and antiviral medicines to workers--such as health
care, transportation, and post office personnel--who are needed to keep
essential services operating. Some say the plan should be more like
that of the U.K., which contains very specific strategies for distributing
medicines and maintaining services during a pandemic. "If you have
antiviral medicines or even a vaccine, you won't have enough. You are
not going to be able to distribute this to everybody. So you need to
have a plan," says David M. Ozonoff, professor of environmental health
at Boston University School of Public Health.

THE VIRUSES involved in all pandemics originate in birds because

birds are the natural reservoir for flu viruses. Since the early 1500s, the