Contents

Abstract
………………………………………………………………….1
1-Introduction
1.1-History
………………………………………………………………….2
1.2-properities and chemistry
……………………………………………………………….....2
1.3-synthesis
………………………………………………………………….3
1.4-production
………………………………………………………………….3
1.5 discussion of influenza virus
……………………………………………………………….4-5
-Miscellaneous
……………………………………………………………….6-8
2-pharmacology
2.1-mechanism of action
…………………………………………………………………9
2.2-pharmacokinetikes
…………………………………………………………………9

1
2.3-pharmacodynamices
………………………………………………………….....9-10
2.4-resistance
………………………………………………………………10
-Miscellaneous
………………………………………………………………11
3-Adminstration and dosing
3.1-posology and method of administration
………………………………………………………………12
3.2-Dosage according to indication
……………………………………………………….…..12-13
3.3-Storage
……………………………………………………………….13
-Miscellaneous
……………………………………………………………….14
4- Contraindication, precaution and side effects
…………………………………………………………...15-16
5-Conclusion
……………………………………………………………...17
6-Reference
……………………………………………………………..18

2
 Abstract
Tamiflu® is a semi synthetic alkaloid that has
antiviral activity. It classified as neuraminidase
inhibitor specifically for influenza A and B. Tamiflu®
contains oseltamivir as phosphate salt.
Oseltamivir derived from many natural sources like
shikimic acid and quinic acid. Shikimic acid is
obtained from star anise fruit (Illicium verum) or
biotechnology. Quinic acid is obtained from cinchona
bark (Cinchona officinalis).
In the pandemic virus of 2009 Tamiflu® plays an
important role in relieve the virulence of the H1N1
virus and save life of thousands of people, however
resistant to the drug is reported Tamiflu® is antiviral
agent that deserves studying and research.
This assay is concerned to overview the properties,
pharmacological and mechanistic of Tamiflu®
especially with the excessive using and dependence of
medical society as the last defense against the
epidemic virus of 2009.

Key words: - Tamiflu®, oseltamivir, shikimic acid,

star anise and influenza

3
 I-Introduction
1.1History [1]
The original synthesis was first reported by Kim et al. (1997). Later,
successful modifications were added by Rohloff et al. (1998), and the
synthesis was reviewed by Abdel-Magid et al. (2001).

It was produced as an attempt to identify potentially orally bioavailable

influenza virus neuraminidase inhibitors, so a series of carbocyclic
transition-state analog inhibitors of the influenza virus neuraminidases
have been designed and synthesized by Kim et al. (1997).

In these compounds, lipophilic side chains replaced the polar glycerol

moiety, and an amino group replaced the guanidino group of the sialic
acid based inhibitor (zanamivir) fig.1.1 in trial to overcome problems of
poor bioavailability.

Although oseltamivir carboxylate is more lipophilic than zanamivir, its

oral bioavailability in rats was still poor.

However, a lack of the polar guanidino and glycerol groups present in

zanamivir and the carboxylic acid group offered an opportunity to make
the molecule orally bioavailable through the prodrug approach.

Ester prodrugs have often been used to increase the bioavailability of

carboxylic acid-containing compounds, once absorbed; these prodrugs are
readily hydrolyzed by a variety of esterases ubiquitously present in the
blood and tissues of many species to release the parent drug that has the
pharmacological activity.

The ethyl ester prodrug of oseltamivir carboxylate appears to satisfy all of

these requirements. After oral administration, oseltamivir is readily
absorbed from the gastrointestinal tract and extensively converted to the
active metabolite, oseltamivir carboxylate.

1.2Properities and chemistry [1]

Oseltamivir fig.1.2 is the pharmacologically active compound in
Tamiflu®. It is an ethyl esterprodrug requiring ester hydrolysis for
conversion to the active form, oseltamivir carboxylate, which is a potent

4
carbocyclic transition-state analog inhibitor of influenza virus
neuraminidase with activity against both influenza A and B viruses in
vitro.

It is white crystalline solid highly water-soluble and non-hygroscopic

prodrug of the active metabolite oseltamivir carboxylate.

1.3 syntheses [1]

In the current commercial method, the starting material in the synthesis of
oseltamivir phosphate is the epoxide, which itself is synthesized in five
steps from either (–)-shikimic acidfig.1.3 or (–)-quinic acidfig.1.4, both of
which are derived from biological sources plant or fermentation origin

Shikimic acid originates from star anisefig1.5 or from fermentation

(using genetically engineered E. coli), and quinic acid is derived from
cinchona bark.fig1.6 There are three chiral centers in the molecule.

The chirality of the epoxide starting material has been confirmed, and
mechanistic, spectroscopic and X-ray structure analyses have shown that
oseltamivir phosphate, as used in the pre-clinical and clinical
development program and marketed, is in the 3R,4R,5S configuration.

1.4 Production [2]

According to Roche (producer company), the major bottleneck in
oseltamivir production is the availability of shikimic acid, which cannot
be synthesized economically and is only effectively isolated from
Chinese star anise.

Unfortunately, the plant takes six years to flower, and it is difficult to

cultivate. One estimate is that 10 years would be needed to produce
sufficient quantities to treat just 20% of the world’s population.

Extraction of shikimic acid from star anise is not easy process that may
take one year, however it considered a safe process if compared with the
conversation of epoxide adduct to azide (which is a reaction that produces
highly explosive material) and limited number of companies around the
world ( Currently only one US company and two European companies)
are approved by the drug regulatory authorities in America and Europe to
carry out the process.

5
Although the fact that star anise is the industrial source for production,
Roche begin to develop alternative methods for synthesize shikimic acid,
especially after the temporary shortage in star anise in 2005.

These alternative methods include organic chemical synthesis (Mirza and

Harvey, 1991; Takeuchi et al., 2000) and methods based on genetically
modified Escherichia coli strains [3] (Chandran et al., 2003; Draths et al.,
1999; Knop et al., 2001).fig.1.7

Also, more safe methods including an azide free process to avoid the
explosion danger, however it is longer (17 steps) and more costive
process or beginning with 5-amino shikimic acid as precursor but using
of this route is more commercially expensive.

See the schematic figure that overall production process fig.1.8

1.5-Discussion of influenza virus [1]

Influenza virus fig.1.9 infections afflicted humans throughout historyand
continue to be a serious health concern. Each year, influenza infects an
estimated 120 million people in the United States, Europe, Japan and
Australia and is a major cause of morbidity and mortality.

In the United States, out of an estimated 20 million cases of flu every

year, up to40, 000 Americans die and more than 400,000 are hospitalized

In periods of flu pandemics, infection rates have been even higher .In
1918–1919, the influenza commonly called" Spanish flu" killed more
than 20 million people with some one billion people affected by the
disease—half of the total human population at the time. New, emerging
influenza viruses such as the H5N1 avian flu and H1N1 swine flu are also
of major concern.

Vaccines provide only partial protection due to their underutilization and

the occasional failure to accurately predict annual changes in the
antigenic determinants of the surface glycoproteins (Couch, 1993).

If the current H1N1 swine influenza outbreak ever assumes the role of a
pandemic, vaccines will become available only after a significant period
of time and then to only a small percentage of the population.

6
 As a result, the use of antivirals could be critical in limiting the initial
spread of a new pandemic. Until recently, only limited options existed for
the control of influenza virus infections.

M2 proton channel blockers (amantadine and rimantadine) have a narrow

spectrum of antiviral activity. They are not active against influenza B or
the H5N1and H1N1 influenza virus

Also, virulent strains resistant to these agents develop quickly (Belshe et

al., 1989). Thus, there was and is an urgent need to identify new antiviral
agents that can be used to prevent and treat influenza virus infections. In
the last few years, new developments in the prevention and treatment of
influenza have been introduced.

The influenza virus neuraminidase (sialidase) has long been considered a

potential antiviral target. This enzyme, which is expressed on the surfaces
of influenza A and B viruses, hydrolyzes terminal sialic acid residues
from glycoproteins, glycolipids, and oligosaccharides.

The influenza virus neuraminidase is required for the aggregation and

release of newly synthesized viral particles from infected cells and is
essential for virus replication (Palese and Compans, 1976).

Influenza virus (NA target site of oseltamivir)

7
 Miscellaneous

Fig.1.1 Structure of carbocyclic sialic acid Fig.1.2 structure of oseltamivir (lipophilic-chain-

analogue zanamivir (less lipophilic –less removal-of guanidino group more lipophilic
bioavailability) properties and formulated in carboxylic form
prodrug-more bioavailability)

Fig.1.3Shikimic acid the main precursor for Fig.1.4Quinic acid an alternate precursor for
production of oseltamivir obtained from production of oseltamivir
star anise or bacterial fermentation or by
organic synthesis

Fig.1.5 star anise the main natural Fig.1.6 cinchona bark source of quinic acid
source to obtain shikimic acid

8
 Fig.1.7The production of shikimic acid in E. coli.

Intermediates(abbreviations): glucose-6-phosphate (G6P), phosphoenolpyruvic

acid(PEP), 3-deoxy-D-arabino-heptulosonic acid (DAHP), 3-dehydroquinic acid
(DHQ), 3-dehydroshikimic acid (DHS), shikimic acid (SA), shikimic acid-3-phosphate
(S3P), quinic acid (QA), gallic acid (GA). Enzymes (encoding genes): (a, b, c) DAHP
synthase (a¼aroF, b¼aroG, c¼aroH). (d) 3-dehydroquinate synthase (aroB), (e) 3-
dehydroquinate dehydratase (aroD), (f) shikimic acid dehydrogenase (aroE), (g)
shikimic acid kinase I (aroK), (h) shikimic acid kinase II (aroL), PPP¼pentose

9
 Commercial Alternative
main source sources

Star Anise E.coli& Organic Cinchona bark

synthesis

Shikimic acid Quinic acid

5-aminoshikimic
acid

Epoxide

Conversation to azide
Explosion hazards (4steps)

Azide free process Azide

(17 steps)

Multistep synthesis
(6steps)

Oseltamivir in form
of Carboxylate

(Prodrug)

Fig.1.8 schematic presentation shows the different routes of

production of oseltamivir.

10
 II-pharmacology [4]
2.1- Mechanism of action
The proposed mechanism of action of oseltamivir is via inhibition of
influenza virus neuraminidase with the possibility of alteration of virus
particle aggregation and release. Fig.2.1

2.2- Pharmacokinetics:
Absorption: - Oseltamivir phosphate is readily absorbed from the GI tract
after oral administration; co-administration with food has no significant
effect on the peak plasma concentration and the area under the plasma
concentration time curve of oseltamivir carboxylate.

Distribution - The binding of oseltamivir carboxylate to human plasma

protein is low (3%). The binding of oseltamivir to human plasma protein
is 42%, which is insufficient to cause significant displacement-based drug
interactions.

Metabolism - Oseltamivir is extensively converted to oseltamivir

carboxylate by esterases located predominantly in the liver fig.2.2.
Neither oseltamivir nor oseltamivir carboxylate is a substrate for, or
inhibitor of, CYP-450 isoforms.

Excretion - Absorbed oseltamivir is primarily (more than 90%)

eliminated by conversion to oseltamivir carboxylate. Plasma
concentrations of oseltamivir declined with a half-life of 1 to 3 hours in
most subjects after oral administration. Oseltamivir carboxylate is not
further metabolized and is eliminated in the urine. Plasma concentrations
of oseltamivir carboxylate declined with a half-life of 6 to 10 hours in
most subjects. Oseltamivir carboxylate is eliminated entirely (more than
99%) by renal excretion. Renal clearance (18.8 L/h) exceeds glomerular
filtration rate.

2.3- Pharmacodynamic properties

Oseltamivir phosphate is a pro-drug of the active metabolite (oseltamivir
carboxylate). The active metabolite is a selective inhibitor of influenza
virus neuraminidase enzymes, which are glycoproteins found on the
virion surface.

11
 Viral neuraminidase enzyme activity is important both for viral entry
into uninfected cells and for the release of recently formed virus particles
from infected cells, and for the further spread of infectious virus in the
body.

Oseltamivir carboxylate inhibits influenza A and B neuraminidases in

vitro.
-Oseltamivir phosphate inhibits influenza virus infection and replication
in vitro.
-Oseltamivir given orally inhibits influenza A and B virus replication and
pathogenicity in vivo in animal models of influenza infection at antiviral
exposures similar to that achieved in man with 75 mg twice daily.

2.4-Resistance:-
Resistance to the drug has occurred infrequently among different viruses
in response to drug treatment, including A H5N1 viruses, but most
notably has emerged among recently circulating A H1N1 viruses and has
spread throughout the population in the absence of drug use [5].

Crystal structures of enzyme–drug complexes, together with enzymatic

properties, of mutants of H5N1 neuraminidase have provided
explanations for high level oseltamivir resistance due to the common
H275Y mutation [5] .fig.2.3

Influenza A virus isolates with reduced susceptibility to oseltamivir

carboxylate have been recovered by serial passage of virus in cell culture
in the presence of increasing concentrations of oseltamivir carboxylate [6].

Genetic analysis of these isolates showed that reduced susceptibility to

oseltamivir carboxylate is associated with mutations that result in amino
acid changes in the viral neuraminidase or viral hemagglutinin [6].

Cross-resistance [6]: Cross-resistance between zanamivir-resistant

influenza mutants and oseltamivir-resistant influenza mutants has been
observed in cell culture.

12
 Miscellenious

Fig.2.1 Neuraminidase inhibitor mechanism of action

Fig.2.2 metabolism of oseltamivir (Pro drug to parent drug)

13
 III-Administration and Dosage

3.1-Posology and method of administration [7]:-

Tamiflu is supplied in two dosage forms
-capsule. fig.3.1
-suspension fig.3.2

Tamiflu capsules and Tamiflu suspension are bioequivalent

formulations. 75 mg doses can be administered as either:-
- One 75 mg capsule or
- One 30 mg capsule plus one 45 mg capsule or
- By administering one 30 mg dose plus one 45 mg dose of suspension.
Adults, adolescents or children (> 40 kg) who are unable to swallow
capsules may receive appropriate doses of Tamiflu suspension.

During situations when commercially manufactured Tamiflu oral

suspension is not readily available, adults, adolescents or children who
are unable to swallow capsules may receive appropriate doses of Tamiflu
by opening capsules and pouring the contents of capsules into a suitable,
small amount (1 teaspoon maximum) of sweetened food product such as
regular or sugar-free chocolate syrup, honey (only for children two years
or older), light brown or table sugar dissolved in water, dessert toppings,
sweetened condensed milk, apple sauce or yogurt to mask the bitter taste.

The mixture should be stirred and the entire contents given to the patient;
it must be swallowed immediately after its preparation.

Oseltamivir may be taken without regard to food. However, when taken

with food, tolerability may be enhanced.

3.2-Dosage according to indication [7]

Treatment of influenza:

- Adults and adolescents 13 years of age and older - The recommended

oral dose of oseltamivir is 75 mg twice daily for 5 days. Begin treatment
within 2 days of onset of symptoms of influenza.
-In children: - The recommended oral dose of oseltamivir oral suspension
for children 1 year of age and older or adults who cannot swallow a
capsule is in fig.3.3

14
Influenza prophylaxis:
The recommended oral dose of oseltamivir for influenza prophylaxis in
adults and adolescents 13 years of age and older following close contact
with an infected individual is 75 mg once daily for at least 10 days.
Therapy should begin within 2 days of exposure.

The recommended dose for prophylaxis during a community outbreak of

influenza is 75 mg once daily.
Safety and efficacy have been demonstrated for up to 6 weeks. The
duration of protection lasts for as long as dosing is continued.

-Children (1 year of age and older) - Prophylaxis in children following

close contact with an infected individual is recommended for 10 days.
Prophylaxis in patients 1 to 12 years of age has not been evaluated for
longer than 10 days' duration. Begin therapy within 2 days of exposure.

The recommended oral dosage of oseltamivir oral suspension for children

1 year of age and older following close contact with an infected
individual is listed in fig.3.4
3.3 – STORAGE OF THE PHARMACY-COMPOUNDED
SUSPENSION [7]:

Refrigeration: Stable for 5 weeks (35 days) when stored in a refrigerator

at 2° to 8°C (36° to 46°F).

Room Temperature: Stable for five days (5 days) when stored at room
temperature, 25°C (77°F).

Note: The storage conditions are based on stability studies of

compounded oral suspensions, using the above mentioned vehicles,
which were placed in amber glass and amber polyethyleneterephthalate
(PET) bottles. Stability studies have not been conducted with other
vehicles or bottle types.

15
 Miscellaneous

Fig.3.1 Tamiflu capsule dosage form 30mg (45.75

Fig.3.2 Tamiflu suspension dosage form 12mg
is also supplied)

Number of capsules
Body Body Number of bottles needed to obtain the
weight weight Recommended needed to obtain the recommended dose for a
(kg) (lbs) dose for 5 days recommended dose 5-day regimen
≤ 15 ≤ 33 30 mg twice daily 1 10 (30 mg)
> 15 to > 33 to 45 mg twice daily 2 10 (45 mg)
23 51
> 23 to > 51 to 60 mg twice daily 2 20 (30 mg)
40 88
> 40 > 88 75 mg twice daily 3 10 (75 mg)

Fig.3.3Oseltamivir Dosing for the Treatment of Influenza in Children

Number of bottles Number of capsules
Body Body Recommended needed to obtain needed to obtain the
weight weight dosage for 10 the recommended recommended dose
(kg) (lbs) days dose for a 10-day regimen
≤ 15 kg ≤ 33 lbs 30 mg once daily 1 10 (30 mg)
> 15 to > 33 to 45 mg once daily 2 10 (45 mg)
23 kg 51 lbs
> 23 to > 51 to 60 mg once daily 2 20 (30 mg)
40 kg 88 lbs
> 40kg > 88 lbs 75 mg once daily 3 10 (75 mg)

Fig.3.4 Oseltamivir Dosing in Influenza Prophylaxis in Children

16
 IV-Contraindication, precaution and side effects
4.1-Contraindiaction and precautions [4]:-
Bacterial infections: Serious bacterial infections may begin with
influenza-like symptoms or may coexist with or occur as complications
during the course of influenza.
Start of treatment: Efficacy of oseltamivir in patients who begin treatment
after 40 hours of symptoms has not been established.
High-risk patients: Efficacy of oseltamivir in subjects with chronic cardiac
disease or respiratory disease has not been established.
Prevention of influenza: Use of oseltamivir should not affect the evaluation
of individuals for annual influenza vaccination in accordance with
guidelines of the Center for Disease Controls and Prevention Advisory
Committee on Immunization Practices. However there are reports that
oseltamivir isn't effective as a prophylactic treatment in case of H5N1 and
H1N1 virus [8].
In that respect, a case report [8] from Vietnam is alerting (Le et al. 2005).
A 14-year-old girl without contact with poultry cared for her 21-year-old
brother, who suffered from an H5N1 infection.
The girl received oseltamivir prophylactically, but nevertheless
contracted the virus apparently from her brother (some viral clones were
sequence identical). She was then treated with oseltamivir and recovered.
However, virologists isolated a drug-resistant H5N1 virus from her that
showed a characteristic neuraminidase mutation that is known to confer
resistance to this drug.

Repeated courses: Safety and efficacy of repeated treatment courses have

not been established.

Hypersensitivity reactions: Rare cases of anaphylaxis and serious skin

reactions, including toxic epidermal necrolysis, Stevens - Johnson
syndrome, and erythema multiforme, have been reported in
postmarketing experiences with oseltamivir. Stop oseltamivir and
institute appropriate treatment if an allergic-like reaction occurs or is
suspected.

Renal function impairment:

Influenza treatment - Dose adjustment is recommended for patients
with creatinine clearance between 10 and 30 mL/min receiving
oseltamivir for treatment of influenza. In these patients, it is
recommended that the dose be reduced to oseltamivir 75 mg once
daily for 5 days.

17
Influenza prophylaxis - For prophylaxis of influenza, dose adjustment is
recommended for patients with creatinine clearance between 10 and 30
mL/min receiving oseltamivir. In these patients, it is recommended that
the dose be reduced to 75 mg every other day.

Pregnancy and Lactation::Tamiflu is categorized in c category according to

F.D.A .It is not known whether oseltamivir or oseltamivir carboxylate is
excreted in human breast milk. Therefore, use oseltamivir only if the
potential benefit for the breast-feeding mother justifies the potential risk
to the breast-fed infant.

Children: The safety and efficacy in children less than 1 year of age have
not been established.

Drug Interactions
Probenecid: Co-administration of probenecid results in an approximate 2-
fold increase in exposure to oseltamivir carboxylate because of a decrease
in active anionic tubular secretion in the kidney.

Adverse Reactions
Adverse reactions occurring in at least 3% of patients include abdominal
pain, cough, diarrhea, fatigue, headache, nausea, and vomiting.

Conclusion
The simple and predictable pharmacokinetic profile of
oseltamivir and convenient oral regimen resulting in effective
antiviral systemic concentrations over the entire dosing interval
made it a useful agent that has become by far the most
frequently prescribed neuraminidase inhibitor for the treatment
and prevention of influenza.

18
Reference
1-Eugene J. Eisenberg, Biotechnology: Pharmaceutical Aspects, Volume
V, Prodrugs Challenges and Rewards Part 1, pages1323-1334, Springer
New York.

2-John W .Frost et al, synthesis of oseltamivir carboxylate, board of

trustees of Michigan State, pub.no.:us2007/0190621 Al, pub. Date
Aug.16, 2007
3-Louise Johansson et al, BIOTECHNOLOGY AND BIOENGINEERING, VOL. 92,
NO. 5, DECEMBER 5, 2005 , Shikimic Acid Production by a Modified Strain of E.
coli, Published online 20 October 2005 in Wiley InterScience

4- Arvind Subramanian MBA, DRUG FACTS & COMPARISONS® POCKET

VERSION REVIEWERS, Chapter 9 - Systemic Anti-Infective Agents,
oseltamivir,pages-1105-1107, Lippincott Williams & Wilkins,2009.

5- P.J. Collins et al, Structural basis for oseltamivir resistance of

influenza viruses, Vaccine 27 (2009) 6317–6323

6- U.S Food and Drug Administration website

http://www.fda.gov

7-Dailymed: -
http://dailymed.nlm.nih.gov/dailymed/drugInfo.cfm?id=11065#HS

8- Harald Brüssow, the Quest for Food A Natural History of Eating, We

as Food and Feeders, chapter7, page 682, Springer New York, 2007.

19