U.S. patent application number 13/523471 was filed with the patent office on 2013-06-13 for topical antiviral formulations.
This patent application is currently assigned to 3B Pharmaceuticals, Inc.. The applicant listed for this patent is Darby C. Brown, Kristen A. Brown. Invention is credited to Darby C. Brown, Kristen A. Brown.
Application Number | 20130150312 13/523471 |
Document ID | / |
Family ID | 45771138 |
Filed Date | 2013-06-13 |
United States Patent
Application |
20130150312 |
Kind Code |
A1 |
Brown; Darby C. ; et
al. |
June 13, 2013 |
TOPICAL ANTIVIRAL FORMULATIONS
Abstract
The disclosure provides antiviral pharmaceutical compositions
comprising one or more antiviral compounds and 2-deoxy-D-glucose in
the form of lip-balms, creams and ointments. A specific embodiment
discloses a lip-balm composition comprising acyclovir and
2-deoxy-D-glucose.
Inventors: |
Brown; Darby C.; (Parker,
CO) ; Brown; Kristen A.; (Parker, CO) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Brown; Darby C.
Brown; Kristen A. |
Parker
Parker |
CO
CO |
US
US |
|
|
Assignee: |
3B Pharmaceuticals, Inc.
Kansas City
KS
|
Family ID: |
45771138 |
Appl. No.: |
13/523471 |
Filed: |
June 14, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
12572584 |
Oct 2, 2009 |
8236768 |
|
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13523471 |
|
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61102729 |
Oct 3, 2008 |
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Current U.S.
Class: |
514/23 |
Current CPC
Class: |
A61K 31/522 20130101;
A61K 31/7004 20130101; A61K 31/522 20130101; A61P 31/12 20180101;
A61K 9/0014 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101;
A61K 31/7004 20130101 |
Class at
Publication: |
514/23 |
International
Class: |
A61K 31/7004 20060101
A61K031/7004; A61K 31/522 20060101 A61K031/522 |
Claims
1. A pharmaceutical composition comprising a therapeutically
effective amount of an antiviral compound selected from one or more
of the group consisting of acyclovir, vidarabine, azidothymidine,
ganciclovir, famciclovir, penciclovir, brivudine, cidofovir,
trifluridine, foscarnet, and a pharmaceutically acceptable salt or
hydrate thereof; and a therapeutically effective amount of
2-deoxy-D-glucose.
2. The composition of claim 1, wherein the antiviral compound is
selected from the group consisting of acyclovir, famciclovir,
penciclovir, and foscarnet.
3. The composition of claim 2 wherein the antiviral compound is
acyclovir.
4. The composition of claim 1 in a form selected from a lip-balm,
stick, cream or ointment.
5. The composition of claim 4 in a lip-balm form.
6. The composition of claim 5, further comprising one or more
polyethylene glycols.
7. The composition of claim 6, further comprising one or more
sweeteners or flavorings.
8. The composition of claim 7 wherein the sweetener is
stevioside.
9. The composition of claim 7 wherein the flavoring is spearmint
oil.
10. The composition of claim 3 wherein the composition comprises
from about 3 wt % to about 7 wt % of acyclovir and from about 0.1
wt % to about 5 wt % of 2-deoxy-D-glucose.
11. The composition of claim 10 wherein the composition comprises
about 5 wt % of acyclovir and about 0.2 wt % of
2-deoxy-D-glucose.
12. A method of treating a herpes viral infection of a patient in
need thereof comprising administering a pharmaceutical composition
comprising a therapeutically effective amount of an antiviral
compound selected from the group consisting of acyclovir,
vidarabine, azidothymidine, ganciclovir, famciclovir, penciclovir,
brivudine, cidofovir, trifluridine, foscarnet, and a
pharmaceutically acceptable salt thereof; and a therapeutically
effective amount of 2-deoxy-D-glucose.
13. The method of claim 12, wherein the antiviral compound is
acyclovir.
14. The method of claim 12, wherein the composition is in a form
selected from a lip-balm, stick, cream or ointment.
15. The method of claim 14, wherein the composition is in a
lip-balm form.
16. The method of claim 13, wherein the composition comprises from
about 3 wt % to about 7 wt % of acyclovir and from about 0.1 wt %
to about 5 wt % of 2-deoxy-D-glucose.
17. The method of claim 16, wherein the topical comprises about 5
wt % of acyclovir and about 0.2 wt % of 2-deoxy-D-glucose.
18. The method of claim 12, wherein the treatment is for latent
infection of herpes simplex type 1 infection.
19. The method of claim 12, wherein the treatment is for latent
infection of herpes simplex type 2 infection.
20. The method of claim 12, wherein the infection is an orofacial
herpes infection.
21. A method of reducing the duration of a herpes viral infection
outbreak in a mammal in need thereof comprising administering a
pharmaceutical composition comprising a therapeutically effective
amount of an antiviral compound selected from the group consisting
of acyclovir, vidarabine, azidothymidine, ganciclovir, famciclovir,
penciclovir, brivudine, cidofovir, trifluridine, foscarnet, and a
pharmaceutically acceptable salt thereof; and a therapeutically
effective amount of 2-deoxy-D-glucose.
22. The method of claim 21, wherein the antiviral compound is
selected from the group consisting of acyclovir, famciclovir,
penciclovir, and foscarnet.
23. The method of claim 21, wherein the antiviral compound is
acyclovir.
24. The method of claim 21 wherein the treatment is for latent
infection of herpes simplex type 1 infection.
25. The method of claim 21, wherein the treatment is for latent
infection of herpes simplex type 2 infection.
26. The method of claim 21, wherein the infection is an orofacial
herpes infection.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. patent application
Ser. No. 12/572,584, filed Oct. 2, 2009, which claims the benefit
of priority to U.S. Provisional Application No. 61/102,729, filed
Oct. 3, 2008, each of which is incorporated herein by reference in
its entirety.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0002] Not applicable.
BACKGROUND OF THE DISCLOSURE
[0003] 1. Field of the Disclosure
[0004] The disclosure provides novel topical antiviral
pharmaceutical compositions comprising an antiviral compound such
as acyclovir and 2-deoxy-D-glucose.
[0005] 2. Description of the Related Art
[0006] Herpes simplex virus (HSV) infections are ubiquitous, with
approximately 80% of the adult population infected with HSV type 1
and approximately 20% of the adult population also infected with
HSV type 2. HSV type 1 is the cause of herpes labialis, also called
orofacial herpes, or cold sores, and HSV encephalitis. HSV type 2
is the primary cause of initial and recurrent genital herpes, and
neonatal HSV. The typical manifestation of a primary HSV-1 or HSV-2
genital infection is clusters of inflamed papules and vesicles on
the outer surface of the genitals resembling cold sores. (Gupta et
al., 2007, "Genital herpes" Lancet 370 (9605): 2127-37).
[0007] Herpes viruses cycle between periods of active
disease--presenting as blisters containing infectious virus
particles--that last 2-21 days, followed by a remission period,
during which the sores disappear. Many HSV infected people
experience recurrence within the first year of infection. During
recurrence fewer lesions are likely to develop, lesions are less
painful, and lesions heal faster, than those occurring during the
primary infection. Subsequent outbreaks tend to be periodic or
episodic, occurring on average four to five times a year when not
using antiviral therapy.
[0008] Treatment of initial HSV infection, and reactivated latent
HSV infection, typically includes topical application of an
antiviral nucleoside composition to lesions at outbreak. Current
prescription topical treatments for herpes labialis include
Zovirax.RTM. cream (5% acyclovir, GlaxoSmithKline/Biovail
Pharmaceuticals, Inc.) which is FDA approved for the treatment of
recurrent herpes labialis (cold sores) in adults and adolescents
(12 years of age and older). Another topical treatment is
Denavir.RTM. (1% penciclovir, Novartis), which is FDA approved for
the treatment of recurrent herpes labialis (cold sores) in adults.
These treatments are noted to inhibit viral replication; shorten
healing time and duration of symptoms; and are soothing. Acyclovir
and penciclovir are antiviral nucleosides. Zovirax.RTM. is applied
five times a day for four days. Denavir.RTM. is applied every two
hours during the day for four days.
[0009] Zovirax.RTM. ointment (5% acyclovir, GlaxoSmithKline/Biovail
Pharmaceuticals, Inc.) is approved for topical administration and
is indicated in the management of initial genital herpes and in
limited non-life-threatening mucocutaneous Herpes simplex virus
infections in immunocompromised patients. Side effects include mild
pain upon application, pruritis, and rash.
[0010] Several patents list acyclovir as an ingredient in topical
formulations for the treatment of herpes simplex genitalis and
herpes labialis.
[0011] Jones and White, U.S. Pat. No. 4,963,555 is listed in the
electronic orange book for Zovirax.RTM. topical cream, 5%
acyclovir. The '555 patent discloses a topical pharmaceutical
formulation for use in treating virus infections of the skin or
mucosa and containing 9-(2-hydroxyethoxymethyl) guanine, or a salt
or ester thereof, which comprises a dispersed oil phase and a
continuous aqueous phase containing therein water, at least 30 wt %
of a polyhydric alcohol and solublized acyclovir.
[0012] Sintov et al., U.S. Pat. No. 5,585,379, disclose antiviral
topical pharmaceutical compositions containing acyclovir dispersed
in an aqueous gel carrier containing a gelling agent; a
water-soluble carboxylic or dicarboxylic acid salt, such as sodium
or potassium oleate; and a polyhydroxy compound such as glycerine,
propylene glycol and polyethylene glycol.
[0013] Kaufman and Faro, 1999, Clin. Obstet. Gynecol., 28 (1):
152-163 provide a review article describing the clinical features
of genital herpes virus infection as well as a discussion of some
of the epidemiologic features which may be related to the increased
frequency with which this disease is being seen and describing
treatment of genital herpes infections.
[0014] Spruance et al., 2002, Antimicrob. Agents Chemother. 46 (7):
2238-2243, provide a description of a placebo-controlled clinical
trial of a 5% acyclovir topical cream for the treatment of herpes
labialis wherein the mean duration of episodes was 4.3 days for
patients treated with acyclovir cream and 4.8 days for those
treated with the vehicle control.
[0015] Certain patents list 2-deoxy-D-glucose as an ingredient in
topical formulations for the treatment of herpes simplex genitalis
and herpes labialis.
[0016] Blough, U.S. Pat. No. 4,603,122, issued Jul. 29, 1986,
discloses a method of treating herpes virus infection by the
administration of 2-deoxy-D-glucose. For example, for herpes
labialis, the topical treatment consisted of two to three drops of
a 50 mM solution of 2-deoxy-D-glucose in sterile anhydrous
glycerol.
[0017] Blough, U.S. Pat. No. 4,315,001, issued Feb. 9, 1982
discloses a method of treating herpes simplex virus by the
administration of 2-deoxy-D-glucose.
[0018] Blough et al., 1979, J. Am. Med. Assoc, 241(26), 2798-280,
describe a clinical study of the treatment of genital herpes
infections with 2-deoxy-D-glucose.
[0019] Patient satisfaction with current remedies, particularly for
HSV-1--herpes labialis, has been poor. Unfortunately, topical
acyclovir therapy lacks efficacy as compared to oral or parenteral
administrations. The drug concentration in the skin after local
application is 2-3 times lower than after given orally (Greg et
al., 1992, J. Invest. Dermatol. 98:856-63). It is speculated that
the lack of efficacy of topical treatments may be related to the
poor water-solubility and lipophilicity of acyclovir, resulting in
its inadequate skin or mucous membrane partitioning ability.
However, it is still considered safer to give the drug locally.
[0020] It is possible that viral resistance to acyclovir should be
considered in patients who show poor clinical response during
therapy. Resistance of HSV can result from qualitative and
quantitative changes in the viral TK and/or DNA polymerase;
particularly noted in immunocompromised patients. Clearly, there is
room for improvement in topical antiviral compositions for the
rapid, efficacious treatment of HSV infections.
[0021] It is herein disclosed that a novel topical formulation
comprising an antiviral compound and 2-deoxy-D-glucose has been
found to be effective in the treatment of HSV infection.
SUMMARY
[0022] The disclosure provides a topical antiviral pharmaceutical
composition comprising one or more antiviral compounds and
2-deoxy-D-glucose. In one embodiment, the antiviral compound is
selected from one or more of acyclovir, vidarabine, azidothymidine,
ganciclovir, famciclovir, penciclovir, brivudine, cidofovir,
trifluridine, and foscarnet; or a pharmaceutically acceptable salt
or hydrate thereof. In a specific embodiment, the antiviral
compound is acyclovir.
[0023] In one embodiment, the topical composition is in a form
selected from a lip-balm, stick, cream or ointment. In one specific
aspect, the topical composition is in a lip-balm form. In one
aspect, the composition further comprises one or more polyethylene
glycols. In another aspect, the composition optionally further
comprises one or more sweeteners or flavorings. In one aspect, the
sweetener is stevioside. In another aspect, the flavoring is
spearmint oil.
[0024] In one embodiment, the composition comprises from about 3 wt
% to about 7 wt % of acyclovir and from about 0.1 wt % to about 5
wt % of 2-deoxy-D-glucose. In one aspect, the composition comprises
about 5 wt % acyclovir and about 0.2 wt % 2-deoxy-D-glucose.
[0025] The disclosure also provides a method of treating a herpes
viral infection of the skin or mucosa of a mammal which comprises
applying to the skin or mucosa a topical composition comprising a
therapeutically effective amount of an antiviral compound, or a
pharmaceutically acceptable salt thereof, and 2-deoxy-D-glucose. In
one embodiment, the method of treating comprises applying to the
skin or mucosa a composition comprising one or more antiviral
compounds selected from the group consisting of acyclovir,
vidarabine, azidothymidine, ganciclovir, famciclovir, penciclovir,
brivudine, cidofovir, trifluridine, and foscarnet; and
2-deoxy-D-glucose. In a specific embodiment, the method of treating
comprises applying to the skin or mucosa a composition comprising
acyclovir and 2-deoxy-D-glucose. In one aspect, the method of
treating involves topical application of the composition in a form
selected from a lip-balm, stick, cream or ointment. In one specific
aspect, the topical composition is in a lip-balm form. In one
aspect, the method of treating involves topical application of the
composition which further comprises one or more polyethylene
glycols. In another aspect, the method of treating involves topical
application of the composition further comprising one or more
sweeteners or flavorings. In one aspect, the sweetener is
stevioside. In another aspect, the flavoring is spearmint oil. In
one aspect the method is for treatment of latent infection of
herpes simplex type 1 infection. In another aspect, the method is
for treatment is for latent infection of herpes simplex type 2
infection.
[0026] In one aspect, the method of treating involves topical
application of the composition which comprises from about 3 wt % to
about 7 wt % of acyclovir and from about 0.1 wt % to about 5 wt %
of 2-deoxy-D-glucose. In one aspect, the composition comprises
about 5 wt % of the acyclovir and about 0.2 wt % of
2-deoxy-D-glucose.
[0027] The disclosure further provides a method of reducing the
duration of a herpes viral infection outbreak of the skin or mucosa
of a mammal which comprises applying to the skin or mucosa a
topical composition comprising a therapeutically effective amount
of an antiviral compound, or a pharmaceutically acceptable salt
thereof, and 2-deoxy-D-glucose. In one aspect the method is for
treatment of latent infection of herpes simplex type 1 infection.
In another aspect, the method is for treatment is for latent
infection of herpes simplex type 2 infection.
[0028] In one embodiment, the method of treating comprises applying
to the skin or mucosa a composition comprising one or more
antiviral compounds selected from the group consisting of
acyclovir, vidarabine, azidothymidine, ganciclovir, famciclovir,
penciclovir, brivudine, cidofovir, trifluridine, and foscarnet; and
2-deoxy-D-glucose. In a specific embodiment, the method of treating
involves topical application of a composition comprising the
antiviral compound acyclovir and 2-deoxy-D-glucose.
BRIEF DESCRIPTION OF THE DRAWINGS
[0029] FIG. 1 shows a concentration/response graph for in vitro
inhibition of HSV-1 infection of adult human dermal microvascular
endothelial cells when treated with acyclovir, 2-deoxy-D-glucose or
an equimolar combination of the two compounds.
[0030] FIG. 2 shows a concentration/response graph for in vitro
inhibition of HSV-2 infection of adult human dermal microvascular
endothelial cells when treated with acyclovir, 2-deoxy-D-glucose or
an equimolar combination of the two compounds.
DETAILED DESCRIPTION
[0031] The disclosure provides a novel topical antiviral
pharmaceutical composition comprising one or more antiviral
compounds and 2-deoxy-D-glucose. The disclosure provides topical
antiviral compositions in the form of a lip-balm, stick, cream,
ointment, lotion, gel, plaster, or pen. A specific embodiment
discloses a lip-balm stick composition comprising acyclovir and
2-deoxy-D-glucose.
[0032] According to one embodiment, the present invention provides
a pharmaceutical composition, which comprises a therapeutically
effective amount of an antiviral compound, or a pharmaceutically
acceptable salt or ester thereof, and 2-deoxy-D-glucose, together
with a pharmaceutically acceptable diluent or carrier.
[0033] The phrase "pharmaceutically acceptable" is employed herein
to refer to those compounds, materials, compositions, and/or dosage
forms which are, within the scope of sound medical judgment,
suitable for use in contact with the tissues of human beings and
animals without excessive toxicity, irritation, allergic response,
or other problem or complication, commensurate with a reasonable
benefit/risk ratio.
[0034] The phrase "pharmaceutically acceptable carrier" as used
herein means a pharmaceutically acceptable material, composition or
vehicle, such as a liquid or solid filler, diluent, excipient,
solvent or encapsulating material. Each carrier must be
"acceptable" in the sense of being compatible with the other
ingredients of the formulation and not injurious to the patient.
Some examples of materials which may serve as pharmaceutically
acceptable carriers include: (1) sugars, such as lactose, glucose
and sucrose; (2) starches, such as corn starch and potato starch;
(3) cellulose, and its derivatives, such as sodium carboxymethyl
cellulose, ethyl cellulose and cellulose acetate; (4) powdered
tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such
as cocoa butter and suppository waxes; (9) oils, such as peanut
oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil
and soybean oil; (10) glycols, such as propylene glycol; (11)
polyols, such as glycerin, sorbitol, mannitol and polyethylene
glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13)
agar; (14) buffering agents, such as magnesium hydroxide and
aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water;
(17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol;
(20) phosphate buffer solutions; (21) silica gel; and (22) other
non-toxic compatible substances employed in pharmaceutical
formulations.
[0035] "Optional" or "optionally" means that the subsequently
described ingredient may or may not be included in disclosed
compositions. "Optionally" is inclusive of embodiments in which the
described ingredient is present and embodiments in which the
described ingredient is not present.
[0036] The use of "wt %" and "w/w" indicates the relative weight
percent of a specified ingredient when compared to the weight of
the total formulation, unless otherwise specified.
[0037] Also included in the family of compounds of the present
invention are the pharmaceutically acceptable salts, and esters
thereof. The term "pharmaceutically acceptable salts" embraces
salts commonly used to form alkali metal salts and to form addition
salts of free acids or free bases. The nature of the salt is not
critical, provided that it is pharmaceutically acceptable. Suitable
pharmaceutically acceptable acid addition salts of compounds of the
present invention can be prepared from inorganic acid or from an
organic acid. Examples of such inorganic acids are hydrochloric,
hydrobromic, hydroiodic, nitric, carbonic, sulfuric, and phosphoric
acid. Appropriate organic acids may be selected from aliphatic,
cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and
sulfonic classes of organic acids, examples of which are formic,
acetic, propionic, succinic, glycolic, gluconic, lactic, malic,
tartaric, citric, ascorbic, glucoronic, maleic, fumaric, pyruvic,
aspartic, glutamic, benzoic, anthranilic, mesylic, salicylic,
p-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic),
methanesulfonic, ethylsulfonic, benzenesulfonic, sulfanilic,
stearic, cyclohexylaminosulfonic, algenic, galacturonic acid.
Suitable pharmaceutically-acceptable base addition salts of
compounds of the present invention include metallic salts made from
aluminum, calcium, lithium, magnesium, potassium, sodium and zinc
or organic salts made from N,N'-dibenzylethyleneldiamine, choline,
chloroprocaine, diethanolamine, ethylenediamine, meglumine
(N-methylglucamine) and procain. All of these salts may be prepared
by conventional means from the corresponding compounds of by
reacting, for example, the appropriate acid or base with the
compounds of the present invention.
[0038] As used herein, the term "pharmaceutically acceptable ester"
refers to esters which hydrolyze in vivo and include, but are not
limited to, those that break down readily in the human body to
leave the parent compound or a salt thereof. Suitable ester groups
include, for example, those derived from pharmaceutically
acceptable aliphatic carboxylic acids, particularly alkanoic,
alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl
or alkenyl moiety advantageously has not more than 6 carbon atoms.
Examples of particular esters include formates, acetates,
propionates, butyrates, acrylates and ethylsuccinates.
[0039] The compositions may be formulated for any route of
administration, in particular for oral, rectal, transdermal,
subcutaneous, intravenous, intramuscular or intranasal
administration. The compositions may be formulated in any
conventional form, for example, as tablets, capsules, caplets,
solutions, suspensions, dispersions, syrups, sprays, gels,
suppositories, patches and emulsions. In certain specific
embodiments, transdermal, topical formulations are disclosed.
[0040] The term "inhibit" or "inhibiting" refers to a statistically
significant and measurable reduction in activity, preferably a
reduction of at least about 10% versus control, more preferably a
reduction of about 20% or more, still more preferably a reduction
of about 30% or more.
[0041] A "therapeutically effective amount" is an amount of a
compound of the present invention or a combination of two or more
such compounds, such as an antiviral compound and
2-deoxy-D-glucose, which inhibits, totally or partially, the
progression of the condition or alleviates, at least partially, one
or more symptoms of the condition. A therapeutically effective
amount can also be an amount that is prophylactically effective.
The amount that is therapeutically effective will depend upon the
patient's size and gender, the condition to be treated, the
severity of the condition and the result sought. For a given
patient and condition, a therapeutically effective amount can be
determined by methods known to those of skill in the art. For
example, in reference to the treatment of a latent herpes viral
infection using the compositions of the present invention, a
therapeutically effective amount refers to that amount of an
antiviral compound which has the effect of (1) reducing the pain,
tingling, burning or itching of the outbreak, (2) reducing the
duration of the outbreak, (3) reducing the recurrence of outbreaks,
and/or, (4) relieving to some extent (or, preferably, eliminating)
one or more other symptoms associated with the outbreak such as,
for example, ulceration, crusting, weeping and scabbing.
[0042] The term "subject", or "patient", refers to an animal, for
example a mammal, who is the object of treatment. Preferably, the
patient is a human. The subject, or patient, may be either male or
female.
[0043] The term "about" when used to refer to a therapeutically
effective amount of an antiviral compound, or another compound,
includes the specified amount .+-.10%.
[0044] HSV-1 and 2 are virtually identical, sharing approximately
50% of their DNA and having over 80% of common antigens. Both types
infect the body's mucosal surfaces, usually the mouth or genitals,
and then establish latency in the nervous system. For both types,
at least two-thirds of infected people have no symptoms, or
symptoms too mild to notice. However, both types can recur and
spread even when no symptoms are present. By the time they are
teenagers or young adults, about 50% of Americans have HSV-1
antibodies in their blood. By the time they are over age 50, some
80-90% of Americans has HSV-1 antibodies. By comparison, almost all
HSV-2 is encountered after childhood, when people become sexually
active.
[0045] HSVs tend to infect cells of ectodermal origin. After direct
exposure to infectious material (i.e., saliva, genital secretions),
initial viral replication occurs at the entry site in the skin or
mucous membrane. After the initial nonspecific inflammatory
response to primary infection, specific antibody response occurs in
a few days, followed by a cellular immune response in the second or
third week. In persons with cellular immune defects, primary HSV
infection can result in life-threatening disseminated disease. In
rare cases, the initial replication may lead to disease and
life-threatening infection (e.g., encephalitis). After retrograde
axonal flow from neurons at the viral point of entry and local
replication, the viral genome becomes latent.
[0046] HSV latency is defined as the ability to remain in a
nonreplicating form in the dorsal root ganglia of the CNS. No viral
particles are produced during latency. A stimulus (e.g., physical
or emotional stress, fever, ultraviolet light) reactivates the
virus in the form of skin vesicles or mucosal ulcers, with symptoms
less severe than those of the primary infection. Latent HSV can be
reactivated from the trigeminal, sacral, and vagal ganglia. Herpes
labialis is the most frequent clinical sign of reactivation of HSV
infection.
[0047] Herpes labialis, also called orofacial herpes, or cold
sores, is most often caused by Herpes Simplex Virus Type 1 (HSV-1).
Reactivation of HSV, predominantly HSV-1, is rarely associated with
systemic signs and symptoms; rather a prodrome of localized pain,
tingling (parasthesia), burning, or itching frequently precede
recurrent orolabial lesions.
[0048] Herpes labialis outbreak proceeds through several stages.
Outbreaks can be triggered by any one of several factors including
stress, sunlight, fatigue, fever, illness, poor diet, food allergy,
and hormonal changes. The prodrome stage may last from a few hours
to a few days and is generally accompanied by a tingling or burning
sensation around the lips or nose. The blister stage occurs within
a day or two of the prodrome stage, there is the first visible sign
of clusters of small blisters. The blister stage is followed by the
weeping/ulcer stage. This stage is characterized by rupture of the
blisters leaving a shallow reddish ulceration. This is the most
painful and contagious stage. Viral shedding occurs generally
during the first 4-5 days of outbreak commencing during the
prodrome stage. The weeping ulcer stage is followed by the crusting
stage. A scab with a brown crust forms. If the scab cracks, the
sufferer will experience itching, burning and bleeding. The healing
stage follows the crusting stage. If a scab has formed, it will
flake off during the healing stage.
[0049] In recurrent orolabial herpetic infection, the lesions tend
to recur at the same site as the original lesions. Pain is most
severe at the onset of infection and diminishes after 4-5 days.
Patients with primary immunodeficiencies, AIDS, malignancy,
malnutrition, or burns and transplant recipients (e.g., bone
marrow, organs) receiving immunosuppressive therapy can have
unusually severe HSV infections. Beginning antiviral treatment when
prodrome is experienced can reduce the appearance and duration of
lesions in some individuals.
[0050] Genital herpes (herpes genitalis) is a sexually transmitted
disease (STD) caused by the herpes simplex viruses type 1 (HSV-1)
or type 2 (HSV-2). Most genital herpes is caused by HSV-2. Most
individuals have no or only minimal signs or symptoms from HSV-1 or
HSV-2 infection. When signs do occur, they typically appear as one
or more blisters on or around the genitals or rectum. The blisters
break, leaving tender ulcers (sores) that may take two to four
weeks to heal the first time they occur. Typically, another
outbreak can appear weeks or months after the first, but it almost
always is less severe and shorter than the first outbreak. Although
the infection can stay in the body indefinitely, the number of
outbreaks tends to decrease over a period of years. As in herpes
labialis, beginning antiviral treatment when prodrome is
experienced can reduce the appearance and duration of lesions.
[0051] The disclosure provides topical antiviral compositions which
reduce both the severity of symptoms and the duration of HSV
infection outbreak. The novel antiviral compositions comprise one
or more antiviral compounds and 2-deoxy-D-glucose for the treatment
of HSV type 1 and HSV type 2 infections. In one aspect, the
disclosure provides a method for reducing reactivation of a latent
infection of herpes viruses in a human comprising topically
administering a pharmaceutical composition comprising a
therapeutically effective amount of an antiviral compound and
2-deoxy-D-glucose.
[0052] Antiviral compounds to be combined with 2-deoxy-D-glucose in
the compositions of the present disclosure are selected from one or
more of but are not limited to, certain antiviral nucleosides
including Acyclovir (9-(2-hydroxyethoxymethyl)guanine;
2-Amino-1,9-dihydro-9-((2-hydroxyethoxy)methyl)-6H-purin-6-one; CAS
No. 59277-89-3; M.W. 225.207 g/mol), Acyclovir sodium
(2-Amino-1,9-dihydro-9-((2-hydroxyethoxy)methyl)-6H-purin-6-one
monosodium salt, CAS No. 69657-51-8; M.W. 247.189 g/mol),
Vidarabine (adenine arabinoside; CAS No. 5536-17-4), Vidaribine
monohydrate (9-beta-D-Arabinofuranosyl-9H-purine-6-amine
monohydrate; CAS No. 24356-66-9); Azidothymidine (AZT, Retrovir,
Zidovudine, 3-azido-3''-deoxythymidine; CAS No. 30516-87-1),
Ganciclovir (9-(1,3-dihydroxy-2-propoxy)methylguanine, DHPG; CAS
No. 82410-32-0), Ganciclovir sodium
(94(2-Hydroxy-1-(hydroxymethyl)ethoxy) methyl)guanine, Sodium;
2-amino-1,9-dihydro-9-((2-hydroxy-1-(hydroxymethyl)ethoxy)methyl)-
-6H-purin-6-one monosodium salt; CAS No. 107910-75-8 or
84245-13-6); Famciclovir (1,3-Propanediol,
2-(2-(2-amino-9H-purin-9-yl)ethyl)-, diacetate (ester); CAS No.
104227-87-4), Penciclovir
(9-(4-Hydroxy-3-(hydroxymethyl)butyl)guanine; CAS No. 39809-25-1),
Brivudine ((E)-5-(2-Bromovinyl)-2'-deoxyuridine; CAS No.
69304-47-8); Cidofovir (Phosphonic acid,
((2-(4-amino-2-oxo-1(2H)-pyrimidinyl)-1-(hydroxymethyl)ethoxy)methyl)-,
(S)--; CAS No. 113852-37-2); Trifluridine (5-Trifluorothymidine;
CAS No. 70-00-8) and any salts or hydrates thereof. The antiviral
nucleosides may be obtained commercially, or produced by synthetic
procedures known in the art, for example, as described in Izawa and
Shiragami, 1998, "Practical syntheses of antiviral nucleosides",
Pure & Appl. Chem., 70(2): 313-318.
[0053] In one embodiment, the topical antiviral compositions
contain from about a total of about 0.1 to about 10 wt % of one or
more antiviral compounds and 2-deoxy-D-glucose. In one aspect, the
topical antiviral composition comprises from about 3 wt % to about
7 wt % of an antiviral compound. In another aspect, the topical
antiviral compound is present from about 0.5 wt % to about 4 wt %
of an antiviral compound. In yet another embodiment, the topical
antiviral compound is present in from about 1% to about 3% weight
compared to the total weight of the formulation.
[0054] Acyclovir is a synthetic purine nucleoside analogue with in
vitro and in vivo inhibitory activity against herpes simplex virus
types 1 (HSV-1), 2 (HSV-2), and varicella-zoster virus (VZV). The
inhibitory activity of acyclovir is highly selective due to its
affinity for the enzyme thymidine kinase (TK) encoded by HSV and
VZV. This viral enzyme converts acyclovir into acyclovir
monophosphate, a nucleotide analogue. The monophosphate is further
converted into diphosphate by cellular guanylate kinase and into
triphosphate by a number of cellular enzymes. In vitro, acyclovir
triphosphate stops replication of herpes viral DNA. This is
accomplished in 3 ways: 1) competitive inhibition of viral DNA
polymerase, 2) incorporation into and termination of the growing
viral DNA chain, and 3) inactivation of the viral DNA polymerase.
The greater antiviral activity of acyclovir against HSV compared to
VZV is due to its more efficient phosphorylation by the viral
TK.
[0055] It is noted in the Zovirax.RTM. (acyclovir) Cream 5%
prescribing information that resistance of HSV to acyclovir can
result from quantitative and qualitative changes in the viral TK
and/or DNA polymerase. Clinical isolates of HSV with reduced
susceptibility to acyclovir have been recovered from
immunocompromised patients, especially with advanced HIV infection.
Most of the acyclovir-resistant mutants isolated thus far from
immunocompromised patients have been TK-deficient mutants, other
mutants involving the viral TK-gene (TK partial and TK altered) and
DNA polymerase have been isolated. TK--negative mutants cause
severe disease in immunocompromised patients and infants. The
possibility of viral resistance to acyclovir should be considered
in patients who show poor clinical response during therapy. A
multi-therapeutic approach to topical antiviral compositions is
herein disclosed as one solution to the problem of resistance. The
disclosure provides compositions comprising one or more antiviral
compounds and 2-deoxy-D-glucose.
[0056] The antiviral action of 2-deoxy-D-glucose has been known for
decades. 2-deoxy-D-glucose (deoxyglucose; 2-deoxy-D-arabino-hexose;
2-deoxy-D-mannose; 2-desoxy-D-glucose; CAS No. 154-17-6; M.W.
164.156 g/mol) is incorporated directly into glycoproteins and
glycolipids and appears to block the cellular synthesis of the
major glycosylated polypeptide of the herpes virus. For example, in
HSV-infected cells treated with 2-deoxy-D-glucose, hematosides are
notably reduced, with an accumulation of precursor molecules,
namely, the ceramide backbone. In addition, 2-deoxy-D-glucose
appears to prevent the synthesis and transport of nonstructural
glycolipids. 2-Deoxy-D-glucose, therefore, is a rational and
effective chemotherapetic agent in the treatment of genital herpes
because of its ability to prevent the synthesis of macromolecules
required for the envelope biogenesis and recognition phenomenon.
2-deoxy-D-glucose may be purchased commercially. In one embodiment,
the topical antiviral compositions contain from about 0.1 to about
10 wt % of 2-deoxy-D-glucose. In one aspect, the topical antiviral
composition comprises from about 0.1% to about 5% of
2-deoxy-D-glucose.
[0057] The compositions of the disclosure contain a weight ratio of
antiviral compound to 2-deoxy-D-glucose from about 1:100 to about
100:1. In one embodiment, the weight ratio of antiviral compound to
2-deoxy-D-glucose is from about 1:30 to about 30:1. In a specific
aspect, the ratio of acyclovir to 2-deoxy-D-glucose is about 25:1.
In one specific aspect, the molar ratio of acyclovir to
2-deoxy-D-glucose is about 18:1. In another specific aspect, the
molar ratio of acyclovir to 2-deoxy-D-glucose is about 1:1.
[0058] In a specific embodiment, the antiviral compound is
acyclovir. In a specific aspect, the topical antiviral composition
comprises from about 3 wt % to about 7 wt % acyclovir. In a
preferred specific aspect, the topical antiviral composition
comprises from about 5 wt % acyclovir. In another specific aspect,
the topical antiviral composition comprises from about 0.1 wt % to
about 5 wt % of 2-deoxy-D-glucose.
[0059] In another embodiment, the disclosure provides a formulation
comprising penciclovir and 2-deoxy-D-glucose. Penciclovir is an
acyclic nucleoside analog available as a 1% topical cream for the
treatment of recurrent herpes labialis (cold sores) in adults
(Denavir.RTM., Novartis). In one aspect, the penciclovir is present
in from about 0.1% to about 5 wt % of the weight of the
formulation. In a specific aspect, the penciclovir is present in
about 1 wt % of the total weight of the formulation. In another
specific aspect, the topical antiviral composition comprises from
about 0.1% to about 5 wt % of 2-deoxy-D-glucose.
[0060] In another embodiment, the disclosure provides a topical
formulation comprising the non-nucleoside antiviral compound
foscarnet and 2-deoxy-D-glucose. Foscarnet is a phosphonic acid
derivative marketed as foscarnet sodium in an injectable
formulation (Foscavir.RTM., AstraZeneca). Foscarnet is an antiviral
medication used to treat herpes viruses, including drug resistant
cytomegalovirus (CMV) and herpes simplex viruses types 1 and 2
(HSV-1 and HSV-2). Topical foscarnet has been used to treat severe
genital ulceration due to acyclovir-resistant HSV-2. See Pechere et
al., Dermatology, 1998; 197:278-280. In one aspect, the foscarnet
is present in from about 1 wt % to about 5 wt % of the total weight
of the formulation. In a specific aspect, foscarnet is present in
about 2.4 wt % of the weight of the formulation. In another
specific aspect, the topical antiviral composition comprises from
about 0.1% to about 5 wt % of 2-deoxy-D-glucose.
[0061] In one embodiment, the topical antiviral composition can
optionally further comprise one or more sweeteners. The optional
sweetener is added to increase patient acceptability and compliance
with the recommended dosing schedule. However, the sweetener may
not be selected from a simple sugar, as the presence of the simple
sugar may interfere with the action of the 2-deoxy-D-glucose. The
sweetener may be selected from a synthetic or natural sweetener,
for example, aspartame, a cyclamate, saccharin, acesulfame salts,
neo-hesperidin dihydrochalcone, sucralose, alitame, astevia,
stevioside, talin, glycerrhizin, thaumatin, xylitol, and mixtures
thereof. The term saccharin as used herein includes saccharin
itself, saccharin acids, and saccharin salts such as sodium
saccharin. In one aspect, the sweetener is stevioside. The
sweetener is optionally present from about 0.1% to about 5 wt % of
the weight of the topical antiviral compositions.
[0062] In another embodiment, the topical antiviral composition can
optionally further comprise one or more flavoring agents. The
optional flavoring agent is added to increase patient acceptability
and compliance with the recommended dosing schedule. The flavoring
agents that may be used include those flavors known to the skilled
artisan, such as natural and artificial flavors. These flavorings
may be chosen from synthetic flavor oils and flavoring aromatics
and/or oils, oleoresins and extracts derived from plants, leaves,
flowers, fruits, and so forth, and combinations thereof.
Non-limiting representative flavor oils include spearmint oil,
cinnamon oil, oil of wintergreen (methyl salicylate), peppermint
oil, clove oil, bay oil, anise oil, eucalyptus oil, thyme oil,
cedar leaf oil, oil of nutmeg, allspice, oil of sage, mace, oil of
bitter almonds, and cassia oil. Also useful flavorings are
artificial, natural and synthetic fruit flavors such as vanilla,
and citrus oils including, without limitation, lemon, orange, lime,
grapefruit, and fruit essences including apple, pear, peach, grape,
strawberry, raspberry, cherry, plum, pineapple, apricot and so
forth. These flavoring agents may be used in liquid or solid form
and may be used individually or in admixture. Commonly used flavors
include mints such as peppermint, menthol, artificial vanilla,
cinnamon derivatives, and various fruit flavors, whether employed
individually or in admixture. Other useful flavorings include
aldehydes and esters such as cinnamyl acetate, cinnamaldehyde,
citral diethylacetal, dihydrocarvyl acetate, eugenyl formate,
p-methylamisol, and so forth may be used. In a specific aspect, the
flavoring is spearmint oil. The flavor is optionally present from
about 0.1% to about 5% by weight of the topical antiviral
composition.
[0063] In a further embodiment, the topical antiviral composition
can optionally further comprise one or more colorants and/or
opacifiers in order to blend with the skin tone of the patient, so
long as the colorant or opacifier does not interfere with the
antiviral action of the formulation. Colorants include such
compounds as, by way of example and without limitation, titanium
dioxide, talc, FD&C Red No. 3, FD&C Red No. 20, FD&C
Yellow No. 6, FD&C Blue No. 2, FD&C Green No. 5, FD&C
Orange No. 5, FD&C Red No. 8, caramel, ferric oxide, other
FD&C dyes, lakes, and natural coloring agents such as grape
skin extract, beet red powder, beta-carotene, annato, carmine,
turmeric, paprika, and other materials known in the art. The amount
of coloring agent used will vary as desired.
[0064] The topical composition carrier formulation should be stable
and pharmaceutically acceptable. The composition should also enable
incorporation of sufficient amounts of the active ingredients to
give the proper penetration characteristics. In addition to
conventional excipient ingredients in lip-balms, sticks, creams,
lotions, gels or ointments, compositions based on phospholipids,
including sphingolipids can be advantageous. Various absorbent
ointment bases, emulsion ointment bases and water soluble ointment
bases and components are known in the art and may be utilized in
the compositions of the present disclosure, for example, as
described in Remington's Pharmaceutical Sciences, Eighteenth Ed.
1990, Mack Publishing Co., Easton Pa., pp. 1311-1314. Ointment
carrier bases may include, but are not limited to, waxes,
petrolatum, esters of fatty alcohols, and saturated fatty acids,
oleic acid, olive oil, paraffin, starch glycerin, lanolin, cetyl
alcohol, glyceryl monostearate, methylparaben, propylparaben,
glycol ethers, polyethylene glycols, polyoxyl 40 stearate, and
polysorbates. The composition may further comprise optional
additional ingredients selected from one or more of a penetration
enhancer, oil, waxy compound, surfactant, stabilizer, gelling
agent, moisturizer, water or a preservative.
[0065] Optional penetration enhancers serve to improve the
absorption across the skin of the antiviral compound. Penetration
enhancers include vitamin E TPGS (Eastman Chemical Company,
Kingsport, Tenn.), and other vitamin E derivatives as described in
U.S. Pat. No. 6,193,985, which is incorporated herein by reference;
and glyceryl monocaprylate/caprate (Cornwell et al. 1998, Int. J.
Pharmaceutics, 171(2): 243-255). Other penetration enhancers are
described in Smith and Maibach (eds.), Percutaneous Penetration
Enhancers, CRC Press, Inc., Boca Raton, F. L. (1995), which surveys
the use and testing of various skin penetration enhancers, and
Buyuktimkin et al., Chemical Means of Transdermal Drug Permeation
Enhancement in Transdermal and Topical Drug Delivery Systems, Gosh
T. K., Pfister W. R., Yum S. I. (Eds.), Interpharm Press Inc.,
Buffalo Grove, I. L. (1997).
[0066] The oils, waxy compounds, gelling agents and surfactants
selected for the formulation and stabilization of these
compositions are those traditionally employed in the dermatological
arts. The optional oils and/or waxy compounds can constitute from
0.5% to 99.9% of the total weight of the composition. The amount of
oil and/or wax depends on the actual form or physical state of the
composition. Exemplary of such oils are mineral oils (petrolatum);
vegetable oils (sweet almond, macadamia, blackcurrant-pip oil);
synthetic oils such as perhydrosqualene, fatty alcohols, acids or
esters (octyl palmitate, isopropyl lanolate, triglycerides
including those of capric/caprylic acids), oxyethylenated or
oxypropylenated fatty esters and ethers; and silicone oils
(cyclomethicone, polydimethylsiloxanes or PDMS) or fluorinated
oils. Exemplary waxy compounds include jojoba oil, paraffin,
carnauba wax and beeswax.
[0067] Exemplary surfactants (emulsifying and coemulsifying)
include the esters of fatty acids and polyethylene glycol (PEG),
esters of fatty acids and glycerol (glyceryl stearate) or esters of
fatty acids and sugar (sorbitan stearate), as well as the
polyoxyethylenated or polyoxypropylenated derivatives thereof,
cyclomethicones and dimethicone copolyols, and also anionic
surfactants (K or Na alkyl phosphate).
[0068] A preferred stabilizer includes glycol stearate or PEG-150
distearate. The stabilizer, when used, is typically present in an
amount from about 0.1 to 5 weight percent of the composition.
[0069] Exemplary gelling agents include modified clays (bentones),
metal salts of fatty acids (aluminum stearate), ethylene/acrylate
copolymers, silicas, polyethylenes, calcium silicates or,
alternatively, ethyl cellulose.
[0070] Preferred moisturizers include wheat protein (e.g.,
laurdimonium hydroxypropyl hydrolyzed wheat protein), hair keratin
amino acids, sodium peroxylinecarbolic acid, panthenol, tocopherol
(Vitamin E), dimethicone, and the like, and mixtures thereof.
Sodium chloride may also be present, particularly when hair keratin
amino acids are included as a moisturizer. Moisturizers, when used,
are typically present in an amount from about 0.01 to 2 weight
percent, preferably about 0.05 to 1.5 weight percent, more
preferably from about 0.1 to 1 weight percent of the
composition.
[0071] The water used is preferably deionized water.
[0072] Preferred preservatives include tetrasodium ethylene-diamine
tetraacetic acid (EDTA), methylparaben, benzophenone-4,
methylchloroisothiazolinone, methylisothiazolinone, and the like,
and mixtures thereof. Preservatives, when used, are typically
present in an amount from about 0.01 to 6 weight percent,
preferably about 0.05 to 4 weight percent, and more preferably from
about 0.1 to 2 weight percent.
[0073] In one embodiment, the disclosure provides a composition
comprising one or more antiviral compounds and 2-deoxy-D-glucose in
a lip-balm stick carrier formulation. In one aspect, the lip balm
stick carrier formulation is comprised of one or more polyethylene
glycols (PEGs). In a specific aspect, the lip-balm stick carrier
formulation is comprised of PEG 1450, PEG 300, silica gel, a
flavoring and a sweetener. The final mixture described above is
poured, while still warm and fluid, into appropriate tubes and
allowed to cool until solid. The resulting lip balm of the present
disclosure is in the form of a stick. However, the lip balm of the
present disclosure can also be marketed in a small wide mouth jar.
One preferred specific embodiment is shown in Example 4.
[0074] In another embodiment, the disclosure provides a composition
comprising acyclovir and 2-deoxy-D-glucose in an aqueous cream
carrier formulation. Preparation of aqueous creams is described,
for example, in U.S. Pat. No. 4,963,555, expired, which is
incorporated herein by reference. In one aspect, the aqueous cream
comprises 5 wt % acyclovir and 0.2 wt % 2-deoxy-D-glucose. A
specific embodiment is shown in Example 5.
[0075] In another embodiment, the disclosure provides a composition
comprising acyclovir and 2-deoxy-D-glucose in an ointment carrier
formulation. In one aspect the ointment carrier formulation
comprises a polyethylene glycol.
[0076] In one embodiment, the disclosure provides a method of
treating an HSV infection outbreak by topically administering a
composition comprising an antiviral compound and 2-deoxy-D-glucose.
A composition of the present disclosure can be administered
topically to the affected area in a single daily dose or in
multiple doses per day. In one aspect, the composition is
administered four times a day. In another aspect, the composition
is administered every three hours during waking hours. In another
aspect, the composition is topically administered every two hours
during waking hours. The treatment regimen may require
administration from a single dose up to multiple daily doses for an
extended period of time, for example, for several days or from one
to two weeks. In one specific aspect, the treatment regimen is
topical application of the composition every 2 hours during waking
hours for 5 days. The amount of antiviral compound and
2-deoxy-D-glucose administered per dose or the total amount
administered will depend on such factors as the nature and severity
of the infection, the age and general health of the patient.
[0077] In one aspect, an equimolar combination of the two compounds
acyclovir and 2-deoxy-D-glucose is unexpectedly more effective than
either individual compound at inhibiting HSV infection of primary
Adult Human Dermal Microvascular Endothelial Cells in vitro. In
vitro assays for anti-HSV-1 and anti-HSV-2 mRNA activity are
described in Example 1. Confluent Vero cell monolayers were
infected with HSV type 1 or HSV type 2 at a multiplicity of
infection of 10 and the resultant virus was used to infect primary
Adult Human Dermal Microvascular Endothelial Cells for 1 hour; the
supernatant was replaced with fresh culture media and then
incubated overnight in the presence of various concentrations of
acyclovir, 2-deoxy-D-glucose or an equimolar combination of the two
compounds. After overnight incubation, the cells were harvested,
lysed, and subjected to QuantiGene Plex 2.0 assay (Panomics, Inc.,
Fremont, Calif.) for quantitative assay of HSV-1 and HSV-2 mRNA.
The QuantiGene Plex 2.0 assay combines branched DNA (bDNA) signal
amplification and multi-analyte profiling beads (xMAP) technologies
to enable the detection and quantitation of multiple mRNA targets
simultaneously.
[0078] FIGS. 1 and 2 show concentration/response graphs for
inhibition of HSV-1 and HSV-2 infection of human endothelial cells
when treated with various concentrations of acyclovir,
2-deoxy-D-glucose or an equimolar combination of the two compounds.
Particularly, FIG. 1 shows a statistically significant leftward
shift of the dose-response curve for the combination of acyclovir
and 2-deoxy-D-glucose (IC.sub.50=1.009 .mu.M) as compared to either
acyclovir (IC.sub.50=1.782 .mu.M) or 2-deoxy-2-D-glucose
(IC.sub.50=20.9 .mu.M). Thus, the combination of acyclovir and
2-deoxy-D-glucose is unexpectedly more effective than either
individual compound at inhibiting HSV infection in vitro. The
combination of acyclovir and 2-deoxy-D-glucose can also be tested
in vitro in either HSV-1 or HSV-2 IgG ELISAs, as described in
Examples 2 and 3.
[0079] In one embodiment, the disclosure provides a method of
treating an HSV infection outbreak by application of the
composition of the present disclosure to a patient in need thereof.
In one aspect, the disclosure provides a method of decreasing the
duration of an outbreak of herpes labialis by application of the
composition of the present disclosure to a patient in need thereof.
In another aspect, application of the composition of the present
disclosure reduces the recurrence of outbreaks of herpes labialis
in a patient in need thereof. In one aspect, the compositions of
the present disclosure reduce the severity of outbreaks of herpes
labialis in clinical patients. In another aspect, the compositions
of the present disclosure reduce the pain associated with outbreaks
of herpes labialis in clinical patients.
[0080] In a preliminary, non-placebo controlled clinical trial,
patients diagnosed with herpes labialis were referred from a
dentist with a prescription for acyclovir. Patients either had an
ongoing outbreak, or were known to be susceptible to recurrent
outbreak following dental work. Patients received the lip-balm
formulation of Example 4 as described in Example 6. Patient surveys
were collected. Data is presented in Table 5. A reduction in the
duration of the outbreak from an average of 10.8 days to 5.2 days
was seen in patients who were administered the formulation of
Example 4. The frequency in the number of annual outbreaks was
reduced from 6.2 to 1.9 per year. Generally, if treatment was
started early at prodrome, no visual outbreak would occur. A
reduction in duration and severity of symptoms was seen if
treatment was started at blister or early crust. Occasionally,
outbreak persisted if treatment was started after prodrome, but an
improvement in symptoms occurred including relief of stinging
and/or reduction in visual severity. The formulation was considered
pleasant.
EXAMPLES
Example 1
In Vitro Assay for Anti-HSV-1 and Anti-HSV-2 mRNA Activity
[0081] Briefly, confluent Vero cell monolayers were infected with
HSV type 1 or HSV type 2 at a multiplicity of infection of 10 and
the resultant virus was used to infect primary Adult Human Dermal
Microvascular Endothelial Cells for 1 hour and then the supernatant
was replaced with fresh culture media and then incubated overnight
in the presence of various concentrations of acyclovir,
2-deoxy-D-glucose or an equimolar combination of the two compounds.
After overnight incubation, the cells were harvested, lysed, and
subjected to QuantiGene Plex 2.0 assay (Panomics, Inc., Fremont,
Calif.) for quantitative assay of HSV-1 and HSV-2 mRNA. The
QuantiGene Plex 2.0 assay combines branched DNA (bDNA) signal
amplification and multi-analyte profiling beads (xMAP.RTM.)
technologies to enable the detection and quantitation of multiple
mRNA targets simultaneously. Generally, this assay is considered to
be a more sensitive and specific assay (bDNA) for the final readout
when compared to an ELISA. The ELISA method while easier, can lack
specificity to determine the difference between HSV1 and HSV2.
[0082] The bDNA assay is a hybridization-based method of
target-specific RNA quantitation that amplifies signal rather than
target RNA, using labeled DNA probes. The QuantiGene Plex 2.0
system utilizes fluorescent microspheres (Capture Beads) as a
support to capture specific RNA molecules. The ability to quantify
multiple target-specific RNA molecules in a single sample lies in
the design of the Probe Sets. For each RNA molecule of interest, an
oligonucleotide Probe Set containing three types of synthetic
probes, Capture Extenders (CEs), Label Extenders (LEs), and
Blockers (BLs) that hybridize and span contiguous sequences of the
target RNA, is provided. The CEs discriminate among the different
Capture Beads within the bead array while capturing, via
cooperative hybridization, the target RNA.
[0083] Signal amplification is mediated by DNA amplification
molecules that hybridize to the tails of the LEs. Each
amplification unit contains multiple hybridization sites for
biotinylated Label Probes that bind Streptavidin-conjugated
R-Phycoerythrin (SAPE). The resulting fluorescence signal
associated with individual Capture Beads is read on a Luminex flow
cytometer. Signal is reported as median fluorescence intensity
(MFI) and is proportional to the number of target RNA molecules
present in the sample. All compounds were run in triplicate for
each concentration. Concentrations are expressed in micromolar
(.mu.M) concentrations.
[0084] The IC.sub.50 (the drug concentration that reduces the
number of virally infected cells by 50%) were determined by
plotting the reduction in fluorescence (RLU) versus the drug
concentration using Graph Pad Prism software. Specifically, the
variable slope model was used to calculate the IC.sub.50s with the
formula "Y=Bottom+(Top-Bottom)/(1+10 ((LogIC50-X)*HillSlope))"
using Graph Pad Prism software. The data is shown graphically in
FIGS. 1 and 2. The error bars represent the standard error of the
mean (SEM). The statistics for each evaluation are shown in Tables
1 and 2.
TABLE-US-00001 TABLE 1 IC.sub.50 of Acyclovir and/or
2-Deoxy-D-glucose against HSV-1. 2 Deoxy D Acyclovir + Nonlin fit
of HSV1 Acyclovir glucose 2DDG log(inhibitor) vs. Variable response
-- slope Best-fit values LogIC50 0.2509 1.32 0.003803 HillSlope
-1.108 -2.395 -1.406 IC50 1.782 20.9 1.009 Std. Error LogIC50
0.05128 0.03338 0.06276 HillSlope 0.1179 0.3951 0.2286 95%
Confidence Intervals LogIC50 0.1463 to 0.3555 1.252 to 1.388
-0.1240 to 0.1316 HillSlope -1.349 to -0.8678 -3.200 to -1.590
-1.871 to -0.9403 IC50 1.401 to 2.267 17.87 to 24.45 0.7517 to
1.354 Goodness of Fit Degrees of Freedom 31 32 33 R.sup.2 0.9715
0.9377 0.9422 Number of points Analyzed 34 35 36
TABLE-US-00002 TABLE 2 IC.sub.50 of Acyclovir and/or
2-Deoxy-D-glucose against HSV-2. 2 Deoxy D Acyclovir + Nonlin fit
of HSV2 Acyclovir glucose 2DDG log(inhibitor) vs. --Variable
response slope Best-fit values LogIC50 0.2805 0.9029 0.212
HillSlope -1.113 -3.957 -1.543 IC50 1.908 7.996 1.629 Std. Error
LogIC50 0.05897 0.03075 0.05944 HillSlope 0.1338 0.8319 0.2717 95%
Confidence Intervals LogIC50 0.1605 to 0.4005 0.8402 to 0.9655
0.09102 to 0.3330 HillSlope -1.386 to -0.8410 -5.652 to -2.262
-2.960 to -0.9895 IC50 1.447 to 2.515 6.922 to 9.237 1.233 to 2.153
Goodness of Fit Degrees of 33 32 33 Freedom R.sup.2 0.9618 0.945
0.9366 Number of points Analyzed 36 35 36
[0085] In these assays, the combination of the two compounds
acyclovir and 2-deoxy-D-glucose was unexpectedly more effective at
inhibiting HSV infection of cells in vitro than either compound
individually, as shown by a comparison of the IC.sub.50 as well as
the concentration/response graphs shown in FIGS. 1 and 2.
Particularly, FIG. 1 shows a statistically significant leftward
shift of the dose-response curve for the combination of acyclovir
and 2-deoxy-D-glucose (IC.sub.50=1.009 .mu.M) as compared to either
acyclovir (IC.sub.50=1.782 .mu.M) or 2-deoxy-2-D-glucose
(IC.sub.50=20.9 .mu.M).
Example 2
HSV-1 IgG ELISA
[0086] The presence of HSV IgG antibody is indicative of previous
exposure. A significant increase in HSV IgG is indicative of
reactivation, current or recent infection. IgM antibody is present
after primary HSV infection. The CALBIOTECH INC. (CBI, Spring
Valley, Calif., Cat. No. H1029G or H1029G4) HSV-1 ELISA Test system
can be used to test for the presence of HSV-1 IgG. A biological
sample is subjected to the HSV-1 ELISA which is an enzyme-linked
immunosorbant assay for the detection of IgG class antibodies to
HSV-1 in human serum. Generally, the CBI HSV-1 IgG ELISA package
insert directions are followed. HSV-1 antigen coated wells are
provided, along with positive and negative controls and
calibrators. For example, diluted patient serum is added to wells
coated with purified antigen. IgG specific antibody, if present,
binds to the antigen. All unbound materials are washed away and the
enzyme conjugate is added to bind to the antibody-antigen complex,
if present. Excess enzyme conjugate is washed off and TMB substrate
is added. The plate is incubated at room temperature for 10 minutes
to allow the hydrolysis of the substrate by the enzyme. Stop
solution is added after 10 minutes. The ELISA plate O.D. is read at
450 nm. The intensity of the color generated is proportional to the
amount of IgG specific antibody in the sample. Generally, an
antibody index <0.9 is interpreted as no detectable antibody to
HSV-1 IgG by ELISA; an antibody index of 0.9-1.1 is a borderline
positive; and an antibody index >1.1 indicates detectable
antibody to HSV-1 IgG by ELISA.
Example 3
HSV-2 IgG ELISA
[0087] The presence of HSV IgG antibody is indicative of previous
exposure. A significant increase in HSV IgG is indicative of
reactivation, current or recent infection. IgM antibody is present
after primary HSV infection. The CALBIOTECH INC. (CBI, Spring
Valley, Calif., Cat. No. H2031G or H2031G4) HSV-2 ELISA Test system
can be used to test for the presence of HSV-2 IgG. A biological
sample is subjected to the HSV-2 ELISA which is an enzyme-linked
immunosorbant assay for the detection of IgG class antibodies to
HSV-2 in human serum or plasma.
[0088] Generally, the CBI HSV-2 IgG ELISA package insert directions
are followed. HSV-2 antigen coated wells are provided, along with
positive and negative controls and calibrators. For example,
diluted patient serum is added to wells coated with purified
antigen. IgG specific antibody, if present, binds to the antigen.
All unbound materials are washed away and the enzyme conjugate is
added to bind to the antibody-antigen complex, if present. Excess
enzyme conjugate is washed off and TMB substrate is added. The
plate is incubated at room temperature for 10 minutes to allow the
hydrolysis of the substrate by the enzyme. Stop solution is added
after 10 minutes. The ELISA plate O.D. is read at 450 nm. The
intensity of the color generated is proportional to the amount of
IgG specific antibody in the sample. Generally, an antibody index
<0.9 is interpreted as no detectable antibody to HSV-2 IgG by
ELISA; an antibody index of 0.9-1.1 is a borderline positive; and
an antibody index >1.1 indicates detectable antibody to HSV-2
IgG by ELISA.
Example 4
Antiviral Topical Formulations
Lip-Balm Formulation.
[0089] Ingredients for one batch of a lip-balm formulation
comprising about 5% w/w of acyclovir and 0.2% w/w 2-deoxy-D-glucose
are provided in Table 3.
TABLE-US-00003 TABLE 3 Lip-balm Formulation. specific Amount per %
Ingredient Amount gravity chapstick (w/w) Acyclovir, USP 1.6667 g
0.333 g 4.5 2-Deoxy-D-glucose 0.0667 g 0.0133 g 0.2 Silica gel,
USP/NF powder 0.200 g 0.04 g 0.5 Stevioside, 90% powder 0.3333 g
0.0667 g 0.9 Polyethylene glycol 1450, 16.25 g 3.25 g 43.6 NF
granules Polyethylene glycol 300, 16.6667 mL 1.1 3.33 mL 49.1 NF
liquid Flavor, spearmint oil 0.5 mL 0.917 0.1 mL 1.2
[0090] The polyethylene 1450 and 300 was melted at 50.degree. C.
with stirring. The acyclovir, 2-deoxy-D-glucose, silica gel and
stevioside were triturated together. The triturated powders were
slowly sifted into the melted PEGs with stirring. The flavoring was
added, followed by thorough mixing. The mixture was poured into
applicator tubes and allowed to cool to room temperature. Amounts
of acyclovir and 2-deoxy-D-glucose employed may be varied in
accordance with the specification.
Example 5
Antiviral Topical Formulations
Aqueous Cream Formulation.
[0091] Ingredients for one batch of an aqueous cream formulation
comprising about 5% w/w of acyclovir and 0.2% w/w 2-deoxy-D-glucose
are provided in Table 4.
TABLE-US-00004 TABLE 4 Aqueous Cream Formulation. % Ingredient
Amount (w/w) Acyclovir, USP 50.0 g 5 2-Deoxy-D-glucose 2.0 g 0.2
Cetostearyl alcohol 67.5 g Sodium lauryl sulphate 7.5 g White soft
paraffin 125.0 g Liquid paraffin 50.0 g Propylene glycol 400.0 g
Purified water, Q.S. to 1000.0 g
[0092] A part of the acyclovir (5 g) is dissolved in the water with
the 2-deoxy-D-glucose and propylene glycol at ambient temperature
to produce an aqueous solution. The paraffins and emulsifiers
(cetostearyl alcohol and sodium lauryl sulphate) are mixed together
and heated to 60.degree. C., and emulsified with the aqueous
solution, also at 60.degree. C., using a laboratory mixer. The
remaining acyclovir is added, the mixture dispersed, allowed to
cool, and filled into lacquered aluminum tubes. Amounts of
acyclovir and 2-deoxy-D-glucose employed may be varied in
accordance with the specification.
Example 6
Clinical Results
[0093] Patients diagnosed with herpes labialis were referred from a
dentist with a prescription for acyclovir. Patients had an ongoing
outbreak, or were known to be susceptible to recurrent outbreak
following dental work. A patient population comprising about 40
patients received the lip-balm formulation of Example 4. Data to
verify treatment outcomes, adverse effects and patient comment was
collected by patient survey several months after dispensing the
formulation. Sixteen patients returned the survey to date. Surveyed
patients ranged in age from 14 to 65 years old, with an average age
of 42 years old. Nine males and seven females returned the survey.
Patients used the formulation for an average of 4.8 days on average
every 2-3 hours following outbreak signs of prodrome or early
crust. Table 5 shows data collected from patient surveys.
TABLE-US-00005 TABLE 5 Clinical Data. Duration of Frequency of
outbreak outbreak Patient Gender Days Number per year No. Age F M
Prior During use Prior During use 1 28 1 0 10 5 12 6 2 14 0 1 12 6
7 2.5 3 44 1 0 10.5 3 6 1 4 53 1 0 14 7 2 1 5 65 0 1 7 5 4.5 0 6 38
0 1 14 9 5 3 7 39 1 0 14 7 2.5 1.5 8 62 0 1 4 2 4 0 9 40 1 0 7 1.5
4 n/a 10 55 0 1 10.5 5 17 1 11 42 1 0 12 6 6 n/a 12 38 0 1 10 6 5
n/a 13 17 0 1 12 8 2.5 n/a 14 48 0 1 11 4 4.5 n/a 15 45 0 1 12 2
4.5 n/a 16 45 1 0 12 7 13 3 Average 42.1 7.0 9.0 10.8 5.2 6.2 1.9
Std. Dev. 14.0 2.8 2.2 4.2 1.9
[0094] A reduction in the duration of the outbreak from an average
of 10.8 days to 5.2 days was seen in all patients surveyed. The
frequency in the number of annual outbreaks was reduced from 6.2 to
1.9 per year. Generally, if treatment was started early at
prodrome, no visual outbreak would occur. A reduction in duration
and severity of symptoms was seen if treatment was started at
blister or early crust. Occasionally outbreak persisted if
treatment was started after prodrome, but an improvement in
symptoms occurred; including relief of stinging, reduction in
visual severity and/or shortened crusting time. Patient
acceptability was good with comments on pleasant smell.
Example 7
Further Clinical Results
[0095] As in Example 5, patients diagnosed with herpes labialis
were referred from a dentist with a prescription for acyclovir.
Patients had an ongoing outbreak, or were known to be susceptible
to recurrent outbreak following dental work. Most patients had
previously been treated with one or more of Zovirax.RTM. (topical
or oral acyclovir), Valtrex.RTM. (oral valacyclovir hydrochloride),
Famvir.RTM. (oral famciclovir), and/or over the counter medications
Abreva.RTM. (topical 10% docosanol), Zilactin.RTM. (topical 10%
benzyl alcohol), Novitra.RTM. (topical zincum oxydatum 2.times.)
and Campho-Phenique.RTM. (topical camphor 10.8% and phenol 4.7%).
The patient population received the lip-balm formulation of Example
4. Data to verify treatment outcomes, adverse effects and patient
comment was collected by patient survey several months after
dispensing the formulation.
[0096] One patient population started application of the topical
test formulation at a prodrome stage comprising tingling and/or
pain, but prior to outbreak. The data for patients who started
treatment by topical application of the test formulation at
prodrome, prior to full outbreak, is shown in Table 6.
TABLE-US-00006 TABLE 6 Clinical Data for Treatment Started at
Prodrome. Product Application Duration of Average outbreak Days
Patient Gender Days During No. Age F M Applied Times/Day Prior Use
1 22 0 1 15.5 5.5 10.5 14 2 48 0 1 9 3.5 4 9 3 51 1 0 1.5 3.5 2 2 4
24 1 0 7.5 5.5 7 6 5 38 0 1 5.5 3.5 7 5 6 50 0 1 7.5 5.5 10 6.5 7
56 1 0 11.5 5.5 14 12 8 20 1 0 7.5 3.5 6 6 9 44 0 1 5.5 3.5 6 1.5
10 53 0 1 3.5 3.5 5 3 11 49 1 0 3.5 3.5 5 1 12 31 1 0 3.5 5.5 7 4
13 36 1 0 1.5 1.5 9 0.5 14 33 0 1 3.5 5.5 10 4 15 45 1 0 1.5 3.5 8
2 16 45 1 0 7.5 5.5 6.5 3 17 52 0 1 3.5 3.5 10 4 18 45 0 1 5.5 5.5
12 2 19 48 0 1 5.5 5.5 4.5 4 20 41 0 1 1.5 1.5 5 3 21 22 0 1 11.5
5.5 14 6 22 59 1 0 3.5 1.5 6 2 23 67 1 0 3.5 3.5 3 3 24 63 1 0 3.5
5.5 7 3 Average 43.4 12.0 12.0 5.6 4.2 7.4 4.4
[0097] When topical treatment with the formulation of Example 4 was
started at prodrome, the average reduction in duration of outbreak
was about three days. Twenty-one of the 24 patients first treated
at prodrome preferred the test formulation to other medications.
All six of the six patients who reported previous use of
Valtrex.RTM., preferred the test formulation. Four of four patients
who reported previous use of Zovirax.RTM. preferred the test
formulation. Patients also generally preferred the test formulation
to Abreva.RTM. (18), Zilactin.RTM. (3), Novitra.RTM. (2), and
Camphophenique.RTM. (1).
[0098] Alternatively, another patient population started the test
formulation at outbreak; or at first blister, weeping or crusting
stage. The data for patients who started treatment by topical
application of the test formulation of Example 4 after outbreak is
shown in Table 7.
TABLE-US-00007 TABLE 7 Clinical Data for Treatment Started after
Outbreak Duration of Product outbreak Application Days Patient
Gender Days During No. Age F M Applied Times/Day Prior use 1 38 0 1
5.5 3.5 7 5 2 55 1 0 7.5 5.5 7 5 3 31 0 1 7.5 3.5 6 7 4 33 1 0 3.5
5.5 7 4 5 13 0 1 3.5 7.5 6 4 6 34 0 1 3.5 5.5 10 6 7 49 1 0 3.5 5.5
6 3 8 31 0 1 3.5 5.5 6 5 9 34 1 0 5.5 5.5 14 6.5 10 25 0 1 7.5 5.5
7 7 11 38 1 0 3.5 5.5 6 3 12 45 0 1 5.5 5.5 12 7 13 54 1 0 9.5 3.5
10 8 14 33 1 0 5.5 5.5 5 2 15 17 0 1 3.5 5.5 12 8 16 38 1 0 3.5 5.5
8.5 3.5 17 18 1 0 5.5 5.5 14 7 18 32 1 0 5.5 5.5 5 3 19 37 1 0 1.5
5.5 11 3 20 9 1 0 3.5 3.5 7 6 21 50 0 1 5.5 1.5 11 6 22 33 1 0 5.5
3.5 6 4 23 53 0 1 5.5 3.5 14 6 24 59 0 1 7.5 3.5 5 3 Average 35.8
13.0 11.0 5.1 4.8 8.4 5.1
[0099] When topical treatment with the formulation of Example 4 was
started after outbreak, the average reduction in duration of
outbreak was about 3.3 days. However, two of 24 patients treated
reported no benefit. Twenty of the 24 patients first treated after
outbreak preferred the test formulation to other medications. Of
four patients that had previously used Valtrex.RTM., three
preferred the test formulation. Of two patients that had previously
used Zovirax.RTM., one preferred the test formulation. Patients
also generally preferred the test formulation to Abreva.RTM. (14),
Zilactin.RTM. (4), Novitra.RTM. (4), and Camphophenique.RTM.
(2).
[0100] In general, regardless of whether the topical test
medication was started at prodrome or after outbreak, patients made
multiple comments that the blisters disappeared, ulcer crusting
time appeared to be shortened, and the tingling and pain stopped.
In addition, very little to no scabbing occurred.
Example 8
Clinical Survey in HSV-2 Patient
[0101] A 23 year old female diagnosed with HSV-2 typically
experienced about one recurrent outbreak per year with prodrome
symptoms of tingling, soreness, and tenderness around the leg.
Outbreak triggers included stress, sunlight, fatigue and hormonal
changes. Typical duration of an outbreak was about 7 days. At the
first signs of outbreak, including tingling and pain, the patient
treated the affected area 3 to 4 times a day for seven days with a
topical stick test formulation comprising 4.5 wt % acyclovir and
0.2 wt % 2-deoxy-D-glucose. The formulation employed was similar to
that of Table 3, but without sweetener or flavoring. The patient
experienced cessation of tingling and pain within 2 days and no
full outbreak occurred. The patient reported that the test
medication suppressed the HSV outbreak better than Valtrex.RTM.
when administered twice a day.
[0102] The various embodiments described above are provided by way
of illustration only and should not be construed to limit the
disclosure. Those skilled in the art will readily recognize various
modifications and changes that may be made to the present
disclosure without following the example embodiments and
applications illustrated and described herein, and without
departing from the true spirit and scope of the present disclosure,
which is set forth in the following claims.
* * * * *