Derivatives of ribavirin

Introduction

Ribavirin is the first synthetic nucleoside with a broad spectrum of antiviral

activity. Ribavirin is a guanosine analog that is phosphorylated intracellularly by host cell
enzymes. Although its mechanism of action has not been fully elucidated, it appears to
interfere with the synthesis of guanosine triphosphate, to inhibit capping of viral messenger
RNA, and to inhibit the viral RNA-dependent polymerase of certain viruses. Ribavirin
triphosphate inhibits the replication of a wide range of DNA and RNA viruses, including
influenza A and B, parainfluenza, respiratory syncytial virus, paramyxoviruses, HCV, and
HIV-1. (1) It is a broad-spectrum virustatic antiviral agent, first synthesised in 1972. (2)

Chemistry

Chemically ribavirin is 1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxamide. (3)

Fig.1.1 Structure of ribavirin

Fig. 1.2 Structure of Ribose

In ribavirin, a 5 membered heterocyclic furan ring is hydroxylated at positions 2 and 3 with a
methanol group at position 4 and a 1 2 4-triazole-3-carboximide ring attached at position 1, a
trait that differentiates it from the ribose sugar which contains a hydroxyl group at position 1.

Mechanism of Action

The exact mechanism of action of ribavirin is poorly understood as of now but there are many
theories regarding it. One such theory states that ribavirin is a guanosine analog and it has
broad-spectrum antiviral activity against both DNA and RNA viruses. It is phosphorylated by
adenosine kinase to the triphosphate, resulting in the inhibition of viral specific RNA
polymerase, disrupting messenger RNA and nucleic acid synthesis. (3) In combination with
pegylated interferon alfa, ribavirin is the standard of care for the treatment of chronic
hepatitis C.

Chemical synthesis of ribavirin

Ribavirin can be synthesized by the reaction of a methyl ester of 1,2,4-triazol-3-carboxylic

acid with O-2,3,5-triacetyl-beta-D-ribofuranose. This reaction yields a methyl ester of 1-O-
2,3,5-triacetyl-beta-D-ribofuranosyl-1,2,4-triazol-3-carboxylic acid, which can be treated
with methanolic ammonia to deacylate the carbohydrate part amidate the carboxyl part to
produce ribavirin. (4)
Derivatives of ribavirin

Many derivatives of ribavirin have been synthesized but only a few of them have shown
potential as medicinal substances. Ribavirin was conceived from the two natural
ribonucleosides, showdomycin and pyrazomycin, both of which show significant broad-
spectrum antiviral activity. (5) Most of the derivatives of this drug are in their early stages of
development and have not been approved for use in humans or animals alike. This is mostly
because of the fact that many of these so-called derivatives are either toxic or do not show
sufficient pharmacological activity. Except for one such derivative, Taribavirin or viramidine,
almost all of the derivatives are experimental in nature and different views and reports exist
regarding their purposed efficacy and/or safety. Ribavirin is associated with significant
haemolytic anaemia, which may require dose reduction, discontinuation or treatment with
recombinant human erythropoietin.

Taribavirin (aka Viramidine)

Fig. 1.3 Structures of ribavirin and taribavirin

The prodrug taribavirin (1-b-D-ribofuranosyl-1H-1, 2, 4-triazole-3-carboxamidine) is a

synthetic nucleoside (guanosine) analog under development for the treatment of patients with
chronic hepatitis C. Taribavirin is metabolized by the liver and converted into its active
metabolite, ribavirin. This pathway reduces exposure to red blood cells (RBCs) and increases
exposure to the liver, the site of HCV replication. (3) As opposed to ribavirin, which
produces haemolysis, taribavirin is supposedly free of such serious side-effects.
It also belongs to the class of organic compounds known as triazole ribonucleosides and
ribonucleotides. These are nucleoside derivatives containing a ribose (or deoxyribose) moiety
which is N-glycosylated to a triazole. Nucleotides have a phosphate group linked to the C5
carbon of the ribose (or deoxyribose) moiety. It is currently in Phase 3 trials. (6)

Advantages of taribavirin (viramidine) when compared to ribavirin

Viramidine is a liver-targeting, synthetic nucleotide prodrug of ribavirin. Viramidine is

predominantly taken up by the liver (first pass effect) and activated (converted) in the liver to
ribavirin by adenosine deaminase. Experimentally, hepatic retention of the ribavirin that is derived
from a single oral dose of viramidine is 3-fold greater than that of oral ribavirin. (7) Viramidine
produces 50% higher levels of ribavirin in the liver but only one-half in the plasma and red
blood cells (RBCs).(8) So, because it produces less concentration of ribavirin phosphates in
the blood or plasma, taribavirin produces considerably less haemolytic anaemia as compared
to ribavirin. Pharmacokinetic and safety studies of viramidine have demonstrated that it is
safe and tolerable. Most reported adverse events are mild. The respective percentages of
treatment-emergent adverse events that were deemed possibly related to viramidine 200, 600
and 1200 mg were 0, 26 and 50%, respectively.(8) The majority of adverse events were mild
and most resolved without consequence.

Chemistry

Taribavirin is quite similar to ribavirin in terms of its chemistry. The only structural
difference between ribavirin and taribavirin being the presence of an amidine group in the
triazole ring at the third position instead of the usual amide group. Being a prodrug,
taribavirin is converted into ribavirin in the liver. This almost always occurs in the liver due
to the higher concentration of hepatic enzymes that can convert amidines into amides.

Chemical synthesis of taribavirin (9)

Fig1.4 Chemical synthesis of taribavirin

Mizoribine (MZB or bredinin)

Mizoribine (MZB) is an imidazole nucleoside and an immunosuppressive agent produced by

the fungus Eupenicillium brefeldianum, an ascomycetes harvested from the soil of Hachijo
Island, Tokyo, Japan, in 1971. It produces mizoribine (MZB). MZB is a nucleoside of the
imidazole class, and was found to have weak antimicrobial activity against Candida albicans.
However, its man use lies in the domain of immunosuppression. This compound is also
known as bredinin and its chemical name is 3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-5-
hydroxyimidazole-4-carboxamide, it is a nucleoside analogue that is structurally similar to
ribavirin.

Chemistry

Fig 1.5 Structure of mizoribine

As figure 1.5 shows, mizoribine is also structurally similar to ribavirin with the changes being
done to the azole ring attached at position 1. Mizoribine contains a diazole ring instead of the
triazole ring and in mizoribine a hydroxyl group is attached to the diazole ring at position 5.

Mechanism of action of mizoribine

Bredinin exerts its immunosuppressive function through selective inhibition of the enzymes
inosine monophosphate dehydrogenase and guanosine monophosphate synthetase, both of
which are required for the generation of guanosine monophosphate from inosine
monophosphate in the de novo pathway. In contrast to azathioprine, bredinin is not
incorporated into nucleic acids in the cells, resulting in fewer side effects, such as
myelosuppression and hepatotoxicity. (10)

Synthesis of mizoribine

Being a natural compound, mizoribine is extracted from the fungus Eupenicillium

brefeldianum.

Merimepodib

Merimepodib (VX-497) is a novel noncompetitive inhibitor of IMPDH. Merimepodib is

orally bioavailable and inhibits the proliferation of primary human, mouse, rat, and dog
lymphocytes at concentrations of approximately 100 nM. (11) This drug is also still in
developmental stages.

Chemistry

This compound belongs to the class of organic compounds known as phenyl-1,3-oxazoles.

These are aromatic heterocyclic compounds containing a 1,3-oxazole substituted at one or
more positions by a phenyl group. The IUPAC name of merimepodib is (3S)-Tetrahydro-3-
furanyl [3-({[3-methoxy-4-(1,3-oxazol-5-yl)phenyl]carbamoyl}amino)benzyl]carbamate. It is
a N-[[3-[[[[3-methoxy-4-(5-oxazolyl)phenyl]amino]carbonyl]amino]phenyl]methyl]-, (3S)-
tetrahydro-3-furanyl ester of carbamic acid.

Fig 1.6 The chemical structure of merimepodib

The compound contains a furan ring like all derivatives of ribavirin. However, the rest of its
structure is quite different from ribavirin. It contains an oxazole ring instead of the usual
triazole ring. An ester linkage is present instead of the amide link and a carbamide linkage is
also present.

Chemical synthesis of merimepodib

Fig 1.7 Chemical synthesis of merimepodib
The oxidation of 3-methoxy-4-methylnitrobenzene (I) with CrO3, H2SO4 and AC2O in
acetic acid gives the gem-diacetate (II), which is hydrolyzed with HCl in refluxing dioxane to
yield 2-methoxy-4-nitrobenzaldehyde (III). Cyclization of (III) with tosylmethyl isocyanide
and K2CO3 in refluxing methanol affords 3-methoxy-4-(5-oxazolyl)nitrobenzene (IV), which
is reduced with H2 over Pd/C in ethyl acetate to provide the corresponding aniline (V). The
activation of (V) with carbonyldiimidazole (CDI) in THF gives the carboxamide (VI), which
is condensed with 3-(tert-butoxycarbonylaminomethyl)aniline (VII), obtained by reaction of
3-aminobenzylamine with Boc2O, and DMAP in refluxing THF to yield the urea (IX).
Deprotection of (IX) with TFA in dichloromethane affords the free benzylamine (X), which
is finally condensed with the 3-furyl ester of the succinimidyl-activated carbonate (XI) by
means of TEA in dichloromethane/DMF. (12)

Mechanism of action of merimepodib

Merimepodib is a orally active inhibitor of inosine monophospate dehydrogenase (IMPDH).

IMPDH inhibition leads to a reduction in intracellular guanosine triphosphate (GTP), a
molecule required for DNA and RNA synthesis. (13) Inosine-5′-monophosphate
dehydrogenase (IMPDH) is a purine biosynthetic enzyme that catalyzes the nicotinamide
adenine dinucleotide (NAD+)-dependent oxidation of inosine monophosphate (IMP)
to xanthosine monophosphate (XMP), the first committed and rate-limiting step towards
the de novo biosynthesis of guanine nucleotides from IMP.
Mycophenolic acid (MPA)

Mycophenolic acid is an antibiotic substance derived from Penicillium stoloniferum. It blocks

de novo biosynthesis of purine nucleotides by inhibition of the enzyme inosine
monophosphate dehydrogenase. Mycophenolic acid is important because of its selective
effects on the immune system. It prevents the proliferation of T-cells, lymphocytes, and the
formation of antibodies from B-cells. It also may inhibit recruitment of leukocytes to
inflammatory sites. It is an an immunosuppresant drug and potent anti-proliferative, and can
be used in place of the older anti-proliferative azathioprine. It is usually used as part of triple
therapy including a calcineurin inhibitor (ciclosporin or tacrolimus) and prednisolone to
prevent organ transplant rejection. It is marketed as mycophenolate mofetil, which is a
prodrug.

Chemistry

This compound belongs to the class of organic compounds known as phthalides. These are
compounds containing a 3-hydrocarbylidene-2-benzofuran-1(3H)-one moiety. Its chemical
name is 6-(4-Hydroxy-6-methoxy-7-methyl-3-oxo-5-phthalanyl)-4-methyl-4-hexenoic acid.

Fig 1.8 the chemical structure of mycophenolic acid

This compound contains the phthalide ring, which is basically a lactone.

Fig 1.9 phthalide ring

Mechanism of action of mycophenolic acid

Mycophenolic acid is a potent, selective, uncompetitive, and reversible inhibitor of inosine

monophosphate dehydrogenase (IMPDH), and therefore inhibits the de novo pathway of
guanosine nucleotide synthesis without incorporation into DNA. Because T- and B-
lymphocytes are critically dependent for their proliferation on de novo synthesis of purines,
whereas other cell types can utilize salvage pathways, mycophenolic acid has potent
cytostatic effects on lymphocytes. Mycophenolic acid inhibits proliferative responses of T-
and B-lymphocytes to both mitogenic and allospecific stimulation. Addition of guanosine or
deoxyguanosine reverses the cytostatic effects of mycophenolic acid on lymphocytes.
Mycophenolic acid also suppresses antibody formation by B-lymphocytes. Mycophenolic
acid prevents the glycosylation of lymphocyte and monocyte glycoproteins that are involved
in intercellular adhesion to endothelial cells and may inhibit recruitment of leukocytes into
sites of inflammation and graft rejection. It is marketed as mycophenolate mofetil in order to
increase its bioavailability. (14)

Fig 1.10 the chemical structure of mycophenolate mofetil

Mycophenolate mofetil is the 2-morpholinoethyl ester of mycophenolic acid. Following oral

and intravenous dosing, mycophenolate mofetil undergoes complete metabolism to MPA, the
active metabolite. Metabolism to MPA occurs presystemically after oral dosing. MPA is
metabolized principally by glucuronyl transferase to form the phenolic glucuronide of MPA
(MPAG) which is not pharmacologically active. In vivo, MPAG is converted to MPA via
enterohepatic recirculation.

Chemical synthesis of mycophenolate mofetil

Mycophenolic acid is used as a starting material, chlorination is performed with a reaction

with thionyl chloride, and then esterification reaction is done with 4-(2-hydroxyethyl)
morpholinem to get mycophenolate mofetil. (15)
Fig 1.11 chemical synthesis of mycophenolate mofetil

EICAR

5-Ethynyl-l-p-D-ribofuranosylimidazole-4-carboxamide (EICAR) is one of the most potent

derivatives of ribavirin. Its antiviral potency is approximately 10 to 100 fold times greater
than that of ribavirin and now it is being evaluated in Phase III clinical trials. The exact
antiviral mechanisms of ribavirin and EICAR are not yet definitely elucidated, although
many hypotheses have been proposed.

Chemistry

EICAR is an imidazole derivative of ribavirin. It also contains an ethyne (alkyne) group

attached to its imidazole ring, which is further attached to the ribofuranose ring at position 1.
Like ribavirin it also contains a amide group attached to the imidazole ring. These features
make it more similar to ribavirin than most other derivatives. It is more lipophilic than
ribavirin due to the introduction of the alkyne group ergo its activity is more pronounced than
ribavirin.
Fig 1.12 the structural similarity between EICAR and ribavirin

Tiazofurin

Tiazofurin is a synthetic nucleoside analogue with antineoplastic activity. Tiazofurin (TR) is

anabolized intracellularly to an analogue of NAD, tiazole-4-carboxamide adeninedinucleotide
(TAD), a potent inhibitor of IMP dehydrogenase (IMPDH); IMPDH is the rate-limiting
enzyme for de novo purine synthesis. Inhibition of IMPDH results in reduced levels of
guanylates, resulting in the inhibition tumor cell growth in vitro and in vivo. Tiazofurin was
rejected for medicinal use due to its severe toxicity. (16)

Fig 1.13 chemical structure of tiazofurin

Discussion
At present, most of the analogues or derivatives of ribavirin are in early research stages.
Many derivatives have been synthesized via chemical reactions by various scientists across
the globe but none have been recognized as medicinal substances as of yet as more research
is need into their pharmacological properties. Derivatives of a certain chemical compound, in
this case ribavirin, may exist but are only considered for further study or research if they
show considerable potential as medicinal substances. If a compound fails to show any
significant pharmacological activity or if it is absurdly toxic at therapeutic doses, it is
discarded and further study or discussion is considered futile.

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