what does the transition state of this reaction look like? note that a key step in the previous...
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What does the transition state of this reaction look like?
• Note that a key step in the previous mechanism is the loss of an alkoxy (sugar) moiety from the position (C2) next to the carboxylate group.
• This creates a carbocation (Sn1 process), which is sp2 hybridized.
• The initial substrate is sp3 hybridized at C2• Thus it was decided to try to synthesize and test
compounds which had a double bond to C2, with the prospect of identifying something which bound tighter than the substrate and which could, therefore, function as an effective inhibitor.
Chemical evolution of neuraminidase inhibitors
• Note that lower Ki values correspond to more active inhibitors• The final product, Relenza (Zanamivir), has a positively charged guanidinium cation (southern end of molecule)• Thus, it is too polar to be absorbed orally and must be administered by inhalation.
Further chemical evolution of neuraminidase inhibitors
While the C5 substituent (glycerol side chain) bound to a polar pocket, some of the more recent analogs have shown there is also a hydrophobic pocket, which can bind more hydrophobic C5 side chains, such as the tertiary amide side chain of I.
• Note that the actual reaction intermediate (lower left) involves a resonance-stabilized carbocation (with oxygen providing electrons to stabilize the C2 carbocation.• Thus it was decided that the double bond of Relenza (Zanamivir) might be in a somewhat incorrect position for optimal binding.• To prepare a stable compound with the double bond in the optimal position, they had to replace the oxygen of the six-membered ring with a carbon (called a carbon isostere).•Note the improvement in activity as the double bond position is altered.
Further chemical evolution of neuraminidase inhibitors
• Thus a series of compounds was prepared having an appropriately placed C=C and an adjacent hydrophobic group (in this case a substituted ether linkage).• The most active of these (above) has a branched side chain (3-pentyl side chain) on the ether.• Due to the improvement in inhibitory activity, it was possible to remove the highly polar guanidinium side chain and replace it with a slightly less polar amine side chain.
Evolution leading to the final product, Tamiflu.
• The carboxylic acid of GS 4071 is still too polar.• Thus, they replaced the acid with an ester, which can be hydrolyzed by esterases.• Compounds related to II (above) are currently in clinical trials. These seem to show a further improvement in selectivity against viruses.
Other Drugs to Treat Influenza
Antiviral Drugs• amantadine (Symmetrel): used
prophylactically against influenza A in high-risk individuals.
• rimantidine (Flumadine): used for treatment and prophylaxis of influenza A.
Mechanism of Action of Amantadine and Rimantadine
• These agents were discovered by random screening and are now known to interfere with a viral ion channed called matriz (M2) protein.
• This causes an inhibition of uncoating of the virus.
• At high concentrations, they also buffer the pH of the endosomes and prohibit the acidic environment needed for Hemaglutanin (HA) to fuse the viral membrane with that of the endosome.
What is HSV?
• HSV = Herpes Simplex Virus• HSV-1 is the ‘usual’ cause of cold sores• HSV-2 is the ‘usual’ cause of genital
herpes• Both types look the same under the
microscope and share about 50% of their DNA.
What is the difference between HSV-1 and HSV-2?
• Both types infect the body’s mucosal surfaces, usually mouth or genitals, and then establish latency in the nervous system.
• For both types, at least two-thirds of the infected people have no symptoms, or symptoms too mild to notice.
• However, both types can recur and spread, even after a period in which there were no symptoms.
http://www.healthscout.com/animation/68/21/main.html
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HSV-1 and HSV-2
The differences• HSV-1 usually establishes
latency in the trigeminal ganglion, a collection of nerve cells near the ear. Then it tends to recur on the lower lip or face.
• HSV-2 usually establishes latency in sacral ganglion at the base of the spine. From there, it recurs in the genital area.
• However, one can have HSV-1 both genitally and orally.
• HSV-1 is usually mild, especially when it infects the lips, face, or genitals.
• However, HSV-1 can recur in the eye, causing ocular herpes, which can lead to blindness, and can even spread spontaneously to the brain, causing herpes encephalitis, which can lead to death.
HSV-1
HSV-2
• 22% of adult Americans have HSV-2• Like HSV-1, HSV-2 symptoms are usually
mild, so mild, in fact, that two-thirds of infected people don’t know they have it.
• HSV-2 rarely causes complications or spreads to other parts of the body.
• Oral HSV-2 infections are rare. But even when an infection does occur, recurrent oral outbreaks are uncommon.
Transmission of HSV-2• In the first year of HSV-2 infection, people shed the
virus from the genital area about 6 to 10% of those days when they are asymptomatic. This decreases over time and can also be further lessened by the use of oral medication. Sex should be avoided in the presence of symptomatic lesions.
• Having a previous HSV-1 infection seems to provide some immunity to an HSV-2 infection. This is probably the reason that oral HSV-2 infections are rare, given the studies which show that a significant proportion of the population practices oral sex.
How severe an infection?• HSV is a lifelong illness• But HSV-2 usually produces only mild symptoms
or signs or no symptoms at all. However, HSV-2 can cause recurrent painful genital sores in many adults, and HSV-2 infection can be severe in people with suppressed immune systems.
• Another factor is how long a person has had the infection. It seems to decrease in severity over time, for reasons which are unclear.
Symptoms
• If signs and symptoms occur during the first episode, they can be quite pronounced. The first episode usually occurs within two weeks after the virus is transmitted, and the sores typically heal within two to four weeks.
• Other signs and symptoms during the primary episode may include a second crop of sores, or flu-like symptoms, including fever and swollen glands.
Is there a cure?
• There is no treatment that can cure herpes, but antiviral medications can shorten and prevent outbreaks during the period of time the person takes the medication.
Vaccines?
• NIH is now in the midst of Phase III clinical trial of an HSV-2 vaccine. This vaccine appears to be about 50% effective.
• If approved, it would be available in 2008.
Antiviral Chemotherapy for HSV
• There are several prescription antiviral medications for controlling herpes outbreaks, include acyclovir (Zovirax), valacyclovir (Valtrex), famcyclovir (Famvir), and pencyclovir.
• Acyclovir was the original and prototypical member of this class
• Valacyclovir and famcyclovir are prodrugs of acyclovir and pencyclovir respectively, with improved oral bioavailability.
Chemotherapy for HSV
Acyclovir (Zovirax) Valacyclovir (Valtrex),
pencyclovir Famcyclovir (Famvir),
O
N
NHN
N
HO
O
H2N
OAc
N
NHN
N
AcO
O
H2N
OH
N
NHN
N
HO
O
H2N
O
N
NHN
N
O
O
H2N
O
H2N
Mechanism of Action of Antivirals to treat HSV
• Both acyclovir and pencyclovir work by interfering with viral replication, effectively slowing the replication rate of the virus, and providing a greater opportunity for the immune response to intervene.
• All drugs in this class depend on the activity of the viral thymidine kinase to convert the drug to a monophosphate form and subsequently interfere with viral DNA replication.
DNA Virus• Recall that HSV is a DNA virus (influenza was an
RNA virus)• In general, more drugs are available to treat DNA
viruses than for RNA viruses (excluding those used to treat HIV).
• Most of the drugs available for treatment of DNA viruses have been developed against herpesviruses.
• Diseases include cold sores, genital herpes, chickenpox, shingles, mononucleosis, etc.
Acyclovir (ZOVIRAX)
• Discovered by random compound screening and introduced into the market in 1981.
• It was the first non-toxic herpes drug to be used systemically.
• It is used for the treatment of infections due to both HSV-1 and HSV-2.
http://www.cat.cc.md.us/biotutorials/dna/dnareppr.html• http://www.dnalc.org/ddnalc/resources/
sangerseq.html
• Aciclovir interferes with DNA synthesis, but must first become activated.
•To become activated, Aciclovir must be phosphorylated (3x)
• However, Aciclovir itself is not a good substrate for mammalian kinases, thus it relies on the viral thymidine kinase to become phosphorylated the first time.
• This is good, since the drug cannot interfere with DNA synthesis in cells that are not infected with the virus, thus reducing the toxicity of the drug.
• The second and third phosphorylations, however, are performed by the cellular thymidylate kinase.
•Aciclovir triphosphate is mistaken for deoxyguanosine triphosphate.
• However, since it lacks the 3’-OH group, it cannot be linked to the adjacent residue in the ‘usual’ fashion.
Valacyclovir
• Valaciclovir is an esterified version of aciclovir, that has greater oral bioavailability.
Penciclovir
• Penciclovir is used primarily as a cream (it has poor oral bioavailability) to treat herpesvirus infections.
• It is the active ingredient in the cold sore medications Denavir and Fenistil..
Famciclovir
• Famciclovir is a di-esterified version of penciclovir• The ester groups give the drug better oral
bioavailability• To treat shingles, it is taken three times a day for
seven days.• To treat genital herpes, it is taken twice a day for five
days.
Major members of the herpesvirus group of the family Herpesviridae
• Herpes simplex viruses: HSV-1 and HSV-2
• Cytomegalovirus (CMV)
• Varicella-zoster virus (VZV)
• Epstein-Barr virus (EBV)
Varicella-zoster virus (VZV)
• The Varicella zoster virus (VZV) is one of the eight herpes viruses known to affect humans (and other vertebrates). It commonly causes chickenpox in children and shingles later in life.
Shingles
Treatment of Varicella Infection• Acyclovir has been shown to reduce fever
and skin lesions in patients with varicella and it use is recommended in healthy patients over 18 years of age.
Acyclovir (ZOVIRAX)
Cytomegalovirus
• Cytomegalovirus (CMV) is a common virus that infects most people during their life. Most people are infected in early childhood (under 3 years of age) or in the teenage years. Usually there are no symptoms of CMV infection.
• Since most CMV infections are mild and usually do not cause long-term problems, most people don't even know they are infected.
• However, CMV can cause problems in a developing baby if the mother gets the infection during pregnancy.
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Cytomegalovirus (CMV)
• After a person has a CMV infection, the virus stays in the body but is not active. The virus can reactivate months or years later, which occurs most often when a person's immune system is weakened.
Cytomegalovirus (CMV)
• Anyone with a weakened immune system is at risk for problems with CMV infection. A weakened immune system can be related to infection with the human immunodeficiency virus (HIV) or medical treatments.
• Medical treatments that can weaken the immune system include: chemotherapy, radiation therapy, steroids, and stem cell or organ transplantation.
Cytomegalovirus (CMV)
• A few people will have symptoms such as sore throat, fever, headache and fatigue. People who have weakened immune systems may develop severe symptoms, such as pneumonia or infections of the eyes, liver, or intestinal tract. People with HIV infection should be sure to let their doctor know if they are having any painless blurring of their vision, "floaters" in the eye, light flashes, areas of blindness, or shortness of breath.
Treatment of CMV Infection• Gangiclovir, a nucleoside analog of acyclovir,
inhibits CMV replication and reduces the severity of CMV syndromes, such as retinitis and gastrointestinal disease.
Deoxyguanosine Acyclovir Gangciclovir
O
OH
N
NHN
N
HO
O
H2N
O
N
NHN
N
HO
O
H2N
O
OH
N
NHN
N
HO
O
H2N
Ganciclovir
Ganciclovir is used to treat or prevent cytomegalovirus
Gangciclovir for ocular use is marketed under the trade name Vitrasert.
Valganciclovir
Valganciclovir is a more orally bioavailable version of this drug.
Epstein-Barr Virus (EBV)
• EBV is the etiologic agent of infectious mononucleosis• Symptoms include fever, tender lymph nodes, sore throat,
mental fatigue• Treatment is usually supportive• Acyclovir can decrease replication of EBV in culture, but has
little impact on clinical illness.
Assigned Reading
• Wasik, Mitzi; Kachlic, Marlowe Djuric. A review of common sexually transmitted diseases. Formulary (2009), 44(3), 78-85.
• De Clercq, Erik; Field, Hugh J. Antiviral prodrugs - the development of successful prodrug strategies for antiviral chemotherapy. British Journal of Pharmacology (2006), 147(1), 1-11.
Homework Questions:1. Describe the methods used for diagnostic testing for HSV.
2. List the accepted treatment for this disease.
3. From one point of view, all antiviral nucleosides can be considered prodrugs. Explain. What enzyme do they require for activation?
4. What two undesirable properties of acyclovir (ACV) prompted the search for a prodrug?
5. What protein is responsible for the improved bioavailability of valaciclovir?
6. Gangiciclovir (GCV) has superior activity than acyclovir against which virus?
7. Oseltamivir (Tamiflu) is also a prodrug. Which functionality will be transformed in vivo? Why was this functionality incorporated?