U.S. patent application number 13/963898 was filed with the patent office on 2013-12-05 for antifungal nail coat and method of use.
This patent application is currently assigned to NexMed Holdings, Inc.. The applicant listed for this patent is Apricus Biosciences, Inc. Invention is credited to Stanley W. Kepka, Mingqi Lu, Y Joseph Mo, William R. Pfister, Hang-Yong Wang.
Application Number | 20130323189 13/963898 |
Document ID | / |
Family ID | 33098141 |
Filed Date | 2013-12-05 |
United States Patent
Application |
20130323189 |
Kind Code |
A1 |
Kepka; Stanley W. ; et
al. |
December 5, 2013 |
ANTIFUNGAL NAIL COAT AND METHOD OF USE
Abstract
Antifungal nail coat compositions containing an allylamine, an
N,N-di(C.sub.1-C.sub.8) alkylamino substituted, (C.sub.4-C.sub.18)
alkyl (C.sub.2-C.sub.18) carboxylic ester or a pharmaceutically
acceptable acid addition salt thereof, a hydrophilic polymer, and a
pharmaceutically acceptable, volatile organic carrier are
disclosed. The composition provides a substantially water-soluble,
fungicidal coating upon contact with a fungally susceptible or
infected nail.
Inventors: |
Kepka; Stanley W.; (Ringoes,
NJ) ; Mo; Y Joseph; (Princeton, NJ) ; Wang;
Hang-Yong; (East Brunswick, NJ) ; Lu; Mingqi;
(Lawrenceville, NJ) ; Pfister; William R.;
(Robbinsville, NJ) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Apricus Biosciences, Inc |
San Diego |
CA |
US |
|
|
Assignee: |
NexMed Holdings, Inc.
East Windsor
NJ
Apricus Biosciences, Inc
San Diego
CA
|
Family ID: |
33098141 |
Appl. No.: |
13/963898 |
Filed: |
August 9, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12316009 |
Dec 9, 2008 |
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13963898 |
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10514190 |
Nov 10, 2004 |
7462362 |
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PCT/US2004/008618 |
Mar 22, 2004 |
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12316009 |
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60456684 |
Mar 21, 2003 |
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Current U.S.
Class: |
424/61 |
Current CPC
Class: |
A61K 47/14 20130101;
A61P 3/02 20180101; A61K 31/137 20130101; A61P 17/00 20180101; A61K
47/32 20130101; A61K 9/08 20130101; A61K 9/0014 20130101; A61Q 3/02
20130101; A61P 31/00 20180101; Y10S 424/808 20130101; A61K 8/41
20130101; A61P 31/02 20180101; A61K 9/7015 20130101; A61P 31/10
20180101; A61K 8/44 20130101 |
Class at
Publication: |
424/61 |
International
Class: |
A61K 31/137 20060101
A61K031/137; A61K 9/00 20060101 A61K009/00 |
Claims
1-22. (canceled)
23. A method for ameliorating or preventing fungal infection of a
toenail or fingernail comprising contacting a fungally susceptible
or infected nail and skin tissue adjacent thereto with a nail coat
composition consisting essentially of, on a total composition
weight basis: about 0.5 to about 10 weight percent terbinafine
hydrochloride: about 0.1 to about 25 weight percent
dodecyl-2-(N,N-dimethylamino) isopropionate hydrochloride: about
0.1 to about 10 weight perecent benzyl alcohol; about 0.1 to about
5 weight percent polyvinylpyrrolidone; and the remainder
ethanol.
24. The method according to claim 23 wherein the contact is
maintained for a period of at least about 0.5 hours.
25. The method according to claim 23 wherein the method is
performed at least once a day.
26. The method according to claim 24 wherein the method is
performed at least once a day.
27. The method according to claim 23, wherein the method is
practiced daily until new nail growth is free of fungal
infection.
28. The method according to claim 24, wherein the method is
practiced daily until new nail growth is free of fungal
infection.
29. The method according to claim 23, wherein the composition is
applied by brushing or spraying.
30. The method according to claim 23, wherein the nail coat
composition consists essentially of, on a total composition weight
basis: about 10 weight percent terbinafine hydrochloride: about 0.5
weight percent dodecyl-2-(N,N-dimethylamino) isopropionate
hydrochloride: about 0.75 weight perecent benzyl alcohol; about 0.5
weight percent polyvinylpyrrolidone; and the remainder ethanol.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation of U.S. patent
application Ser. No. 10/514,190, filed Nov. 10, 2004, now U.S. Pat.
No. 7,462,362, which is a U.S. National Stage of international
application No. PCT/US2004/008618, filed Mar. 22, 2004, which
application claims benefit of U.S. Provisional Patent Application
No. 60/456,684, filed Mar. 21, 2003, incorporated herein by
reference.
TECHNICAL FIELD OF THE INVENTION
[0002] This invention relates to topical antifungal compositions
useful for ameliorating or preventing onychomycoses of the toenails
or fingernails, as well as adjacent skin. More particularly, the
invention relates to a dual action antifungal nail coat composition
and the method for applying the antifungal composition to a
fungally susceptible or infected nail and/or adjacent skin.
BACKGROUND OF THE INVENTION
[0003] Superficial fungal infections of skin, hair, nails, or
mucous membranes, are still very common among all populations. In
particular, onychomycosis is a fungal infection of the nails. The
onychomycosis are frequent, involving up to about 15% of persons
between the ages of 40 and 60 years. Some estimates suggest that
onychomycosis affects about 6 to about 13% of the North American
population, with an estimated 4.9 to 12.3 million people being
affected in the United States. In European populations, the
estimated overall prevalence of onychomycosis is in the range of
about 3 to about 10%.
[0004] Delivery of antifungal agents through the nail into the nail
bed and surrounding skin has been minimally effective for the
treatment of onychomycosis (infections of the fingernails,
toenails, and immediate adjacent surrounding skin) in the form of a
nail lacquers, primarily because the film forming water-insoluble
polymers used limit the diffusion of the drug from the dried film
into the nail and skin, and because previous nail lacquer
compositions do not contain the optimum balance of permeation
enhancement to deliver the drug to both the nail and surrounding
skin in an amount sufficient for optimal antifungal activity.
[0005] Fungal infection of the nails, commonly referred to as
onychomycosis, is most frequently caused by dermatophytes, but can
also be caused by molds and Candida sp. Onychomycosis is
predominantly present in toenails rather than fingernails, in
males, and in the elderly. Onychomycosis is most commonly caused by
Trichophyton rubrum (T. rubrum), Trichophyton mentagrophytes (T.
mentagrophytes), and Epidermophyton floccusum (E. floccusum).
Onychomycosis due to nondermatophytes is usually caused by Candida
species, such as Candida albicans, and is more likely to cause
invasive nail disease in fingernails than in toenails of
immunocompetent individuals.
[0006] Onychomycosis has medical significance especially in
individuals having certain diseases, such as diabetes and others
where the individual is immunocompromised. Also onychomycosis can
have a substantial undesirable effect on daily living activities,
such as ambulation, and spontaneous remission is rare. The current
treatments of onychomycosis include oral administration of
antifungal agents, such as itraconazole (distributed under the
tradename, SPORONOX.RTM., by Ortho Biotech Products L.P.), and
terbinafine (distributed under the tradename, LAMISIL.RTM., by
Novartis Pharmaceuticals Corporation). While itraconazole and
terbinafine hydrochloride offer significant cure rates, shorter
treatment regimens and lower levels of adverse events compared to
the imidazoles (e.g., ketoconazole), clinically significant drug
interactions can occur and the therapeutic period requires at least
a few months. Thus, there is an ongoing need and desire for a
non-oral management of onychomycosis.
[0007] One attempt has been made employing the antifungal agent,
ciclopirox distributed commercially under the trade name PENLAC.TM.
Nail Lacquer by Dermik Laboratories, Inc.), as an 8% topical
solution containing a water-insoluble, film-forming polymer, and is
described in U.S. Pat. No. 4,957,730 to Bohn, et al. Another
antifungal nail lacquer compositions utilizing a water-insoluble
film forming polymer is described in U.S. Pat. No. 6,495,124 by
Samour. Yet another nail lacquer formulation contains 5%
amorolfine, a morpholine derivative, and is manufactured by Roche
Laboratories under the trade name LOCERYL.TM.. However,
water-insoluble film-forming polymers, such as used in conventional
nail lacquer compositions are fast drying (less than one minute)
solution of water-insoluble polymers and, if brushed onto the skin
area surrounding the nail, tend to irritate the skin area.
Additionally, such traditional, water-insoluble, fast drying,
film-forming polymers produce high viscosity nail lacquer
compositions and thus limit the mobility and time for active
exchange of the antifungal agent between the film and the nail
plate resulting in loss of treatment efficacy. In some instances,
the nail lacquers are suitable only for treatment of mild
onychomycosis without nail matrix involvement, and systemic
treatment is still required for severe onychomycosis involving the
nail bed.
[0008] An attempt employing azole derivatives at 0.5-1%
concentration applied from a composition containing water-insoluble
fatty components, solubilizers and a quick drying, water-soluble,
polyvinylpyrrolidone, or vinylacetate copolymers and terpolymers
thereof, is described in Canadian Patent No. 1,175,355 and European
Patent No. 055,397.
[0009] The present dual action antifungal topical nail coat
compositions and methods provide a fungicidal regimen suitable for
the treatment of onychomycosis of varying severities in mammals in
need of such treatment.
SUMMARY OF THE INVENTION
[0010] A dual action antifungal nail coat composition containing an
antifungal agent for ameliorating or preventing fungal infections
of nails and surrounding skin, and onychomycosis in particular, is
disclosed. The present composition delivers the active ingredient
both through the nail plate as well as through the surrounding skin
tissue. Also disclosed are methods for topically applying the dual
action antifungal nail coat composition to a fungally-susceptible
or infected nail. The bioavailability of the antifungal agent is
optimized by the practice of the present invention.
[0011] The antifungal nail coat compositions can be formulated as
"one-coat" type and "two-coat" type compositions.
[0012] A preferred one-coat type antifungal nail coat composition
embodiment comprises: [0013] an effective fungicidal amount of an
antifungal agent; [0014] a permeation enhancing amount of a
substantially non-volatile, permeation enhancer selected from the
group consisting of an N,N-di(C.sub.1-C.sub.8) alkylamino
substituted, (C.sub.4-C.sub.18) alkyl (C.sub.2-C.sub.18) carboxylic
ester or pharmaceutically acceptable acid addition salt thereof, a
pharmaceutically acceptable alcohol, and mixtures thereof; [0015] a
film-forming amount of a hydrophilic polymer; and [0016] a
pharmaceutically acceptable, volatile organic carrier.
[0017] In a one-coat type, dual action antifungal nail coat
composition, the organic carrier preferably assists in distributing
the drug, i.e., the antifungal agent, substantially uniformly on
contact of the nail coat composition with a fungally susceptible or
infected nail and or adjacent skin and volatilizes, within about
one to five minutes following application to provide a
substantially water-soluble, fungicidal film coating on the nail
and adjacent skin tissue containing the drug and one or more
substantially non-volatile penetration enhancer for on-going
amelioration or prevention of fungal infection.
[0018] Another preferred dual action antifungal nail coat
composition is a two-coat type composition comprising: [0019] a
first antifungal nail coat composition for providing an antifungal
primer coat, comprising an effective fungicidal amount of
antifungal agent dispersed in a pharmaceutically acceptable,
volatile organic carrier; [0020] a second antifungal nail coat
composition for providing an antifungal film coat, comprising a
film-forming amount of hydrophilic polymer, an effective fungical
amount of antifungal agent, and a pharmaceutically acceptable,
volatile organic carrier, and [0021] wherein either one of the
first or second antifungal nail coat composition optionally
includes a substantially non-volatile permeation enhancer.
[0022] In a two-coat type, dual action composition, the antifungal
agent is quickly released from the first antifungal nail coat
composition to a fungally-susceptible or infected nail on contact
therewith to provide a fungicidal primer coat. The second
antifungal nail coat composition provides a fungicidal film coat
over the foregoing fungicidal primer-coated nail on subsequent
contact therewith. The fungicidal film coat provides a depot for
additional antifungal agent which can be released over an extended
time period and provides a protective nail barrier to maintain
sustained release of antifungal agent from the primer coat to the
nail to optimize the topical bioavailability of antifungal agent,
and to minimize further accessibility of fungal spores from the
environment to the infected nail.
[0023] The antifungal agent is preferably selected from the group
consisting of allylamine and azole antifungals. The allylamine
antimycotic terbinafine, usually as terbinafine hydrochloride, is
particularly preferred. The azole antifungals include azoles,
imidazoles, as well as triazoles.
[0024] The hydrophilic polymer may be a film-forming polymer
comprising a vinylpyrrolidone monomer unit, including homopolymers
(such as, polyvinylpyrrolidone), copolymers, and complexes thereof,
a gum, a resin, or the like. Preferably, the hydrophilic polymer is
polyvinylpyrrolidone (PVP).
[0025] The volatile organic carrier preferably is a
pharmaceutically acceptable aliphatic alkanol having 2 to about 5
carbon atoms, and more preferably is ethanol.
[0026] Particularly preferred substantially non-volatile permeation
enhancers are dodecyl-2-(N,N-dimethylamino) isopropionate (DDAIP),
benzyl alcohol and combinations thereof.
[0027] The dual action antifungal nail coat compositions of this
invention can include one or more penetration enhancers in an
amount effective to achieve an antifungal concentration of the
antifungal drug in the nail and surrounding skin, as well as an
auxiliary anti-infective, such as an antibacterial agent, an
antiseptic agent, and the like, to augment the efficacy of the
treatment.
[0028] Fungal infection of a toenail or fingernail may be
ameliorated or prevented by fungicidal regimens in which the
disclosed dual action antifungal topical nail coat compositions are
applied in the form of either a one-coat type or a two-coat type by
the methods described herein. The methods of this invention are
preferably performed at least once a day for as long as needed to
ameliorate or prevent fungal infection.
[0029] The practice of this invention using dual action antifungal
nail coat compositions is desired for increasing the topical
bioavailability of an antifungal drug, especially in the treatment
of onychomycoses of the toenails or fingernails. Beneficially, the
dual action antifungal nail coat composition can shorten the total
therapeutic period, avoid or eliminate adverse systemic events
usually associated with oral therapies, and improve clinical
efficacy.
BRIEF DESCRIPTION OF THE DRAWINGS
[0030] In the drawings,
[0031] FIG. 1 is a graphical representation of terbinafine uptake
by human nail clippings from a selected individual expressed as the
concentration of terbinafine remaining in a terbinafine source
solution as a function of time, wherein Solution A was 10 weight
percent terbinafine hydrochloride in anhydrous ethanol, Solution B
was 10 weight percent terbinafine hydrochloride in anhydrous
ethanol plus 10 weight percent of polyvinylpyrrolidone (K-30),
Solution C was 10 weight percent terbinafine hydrochloride in
anhydrous ethanol plus 0.5 weight percent DDAIP.HCl, and Solution D
was 10 weight percent terbinafine hydrochloride in anhydrous
ethanol plus 1 weight percent DDAIP.HCl;
[0032] FIG. 2 is a graphical representation of terbinafine release
from the human nail clippings of the selected individual of FIG. 1
expressed as a calculated amount of terbinafine hydrochloride
remaining in the nail clippings as a function of time, wherein
Sample A was previously treated with Solution A, Sample B was
previously treated with Solution B, Sample C was previously treated
with Solution C, and Sample D was previously treated with Solution
D;
[0033] FIG. 3 is a graphical representation of the calculated
amount of terbinafine hydrochloride retained in human nail
clippings as a function of DDAIP.HCl concentration in a solution of
terbinafine hydrochloride in anhydrous ethanol; and
[0034] FIG. 4 is a graphical representation of the permeation of
terbinafine hydrochloride in human nail clippings as a function of
time from a 10 weight percent solution of terbinafine hydrochloride
in anhydrous ethanol and from a 10 weight percent solution of
terbinafine hydrochloride in anhydrous ethanol and also containing
0.5 weight percent DDAIP.HCl.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0035] The term "dual action" as applied to antifungal nail coat
compositions of this invention means that the nail coat composition
provides a water-soluble, fungicidal film coating that contains the
antifungal drug on the nail and adjacent skin tissue, and a
substantially non-volatile penetration enhancer that promotes the
penetration of antifungal drug into the nail as well as surrounding
skin tissue.
[0036] There is no particular limitation on the antifungal agents
useful for the dual action antifungal nail coat compositions of
this invention, as long as the antifungal agent is effective
against fungi known to cause infections of the toenails or
fingernails, as well as surrounding skin, and onychomycosis, in
particular. A listing of antifungal agents, without limitation
thereto, may be found, for example, in the Thirteenth Edition of
The Merck Index (2001) under the headings "Antifungal (Antibiotic)"
and "Antifungal (Synthetic)" in the Therapeutic Category and
Biological Activity Index section incorporated herein by
reference.
[0037] Suitable antifungal agents include, for example allylamines,
such as terbinafine, naftifine and butenafine; and azoles, such as
imidadazoles and triazoles, and the like. Imidazoles include
ketoconazole, bifonazole, butoconazole, chlordantoin,
chlormidazole, cloconazole, clotrimazole, econazole, enilconazole,
fenticonazole, flutrimazole, isoconazole, lanoconazole, miconazole,
neticonazole, omoconazole, oxiconazole nitrate, sertaconazole,
sulconazole, and tioconazole. Triazoles include fluconazole,
itraconazole, posaconazole, saperconazole, terconazole and
voriconazole. Particularly preferred is the allylamine,
terbinafine; the imidazole, ketoconazole; and the triazoles
fluconazole, and itraconazole.
[0038] This invention as described is particularly applicable to
terbinafine and its acid addition salts without limitation thereto.
The practice of this invention using terbinafine is desired since
increasing the topical bioavailability of this antifungal drug is
useful in the treatment of onychomycosis of the toenails or
fingernails. Beneficially, the dual action antifungal nail coat
composition can shorten the total therapeutic period, avoid and
eliminate systemic adverse events, and improve clinical efficacy
because it is applied to the target site of the fungal
infection.
[0039] Terbinafine is designated chemically as
(E)-N-(6,6-dimethyl-2-hepten-4-ynyl)-N-methyl-1-naphthalene
methanamine and has the following structural formula:
##STR00001##
[0040] The term "terbinafine" as used herein includes the free base
form of this compound as well as chemotherapeutically acceptable
acid addition salts thereof. Suitable salt forms include
hydrochloride, hydrogen fumarate or naphthaline-1,5-disulphonate.
For purposes of the present invention, the inorganic acid salt,
terbinafine hydrochloride, is particularly preferred. Terbinafine
hydrochloride is a synthetic antimycotic allylamine related to
naftifine, and is the active ingredient (equivalent to 250 mg base)
of a commercial antifungal medication sold under the name
LAMISIL.RTM. (Novartis Pharmaceuticals Corporation) formulated in
tablets for oral administration. The preparation of propenylamines,
which includes terbinafine, is described in U.S. Pat. No. 4,755,534
and topical application dosage forms for pharmaceutical use
reported therein are ointments or creams at concentrations of from
0.05 to 5, and 0.1 to 1 weight percent, in particular.
[0041] The present invention permits optimization of the clinical
and mycological efficacy of terbinafine in a comprehensive
management program based on topical treatment for ameliorating
various severities of onychomycosis of fingernails and toenails as
well as the surrounding skin where dermatophytes harbor. The
comprehensive management program preferably comprises a daily
regimen of topically applying a dual action antifungal nail coat
composition as described below to ameliorate or prevent
onychomycosis, and preferably includes removal of the unattached
infected nail at least monthly.
[0042] The dual action antifungal nail coat composition embodiments
can be formulated as a "one-coat" type composition or as a
"two-coat" type composition. The term "one-coat type composition,"
as used herein, means that the antifungal nail coat composition
contains a volatile carrier to assist in initially distributing the
drug on contact with the nail and the surrounding skin, and then
volatilize relatively quickly, (i.e., within a period in the range
of about 0.5 to about 10 minutes), so that the hydrophilic polymer
and substantially non-volatile permeation enhancer can provide a
substantially uniform fungicidal film coat on the nail and adjacent
skin tissue as a depot for the drug to provide ongoing amelioration
or fungicidal prevention efficacy. The fungicidal film coat thus
remains in contact with the nail until the nail coat is removed,
such as by water rinsing or bathing. In this manner, an undesirable
build-up of polymeric carrier films encountered by prior art
antifungal nail lacquers is avoided.
[0043] A one-coat type composition is preferably applied at least
once daily, as needed, and can be re-applied with or without an
intervening water rinse.
[0044] In a one-coat type of antifungal nail composition, the
amount of antifungal agent present usually is in the range of about
0.1 to about 20 weight percent, preferably in the range of about
0.5 to about 15 weight percent, and most preferably in the range of
about 1 to about 5 weight percent.
[0045] The term "two-coat type antifungal nail composition," as
used herein, refers to a two-part, dual action antifungal nail coat
composition formulation, each of which is sequentially applied, at
least once daily. Thus a two-coat type antifungal nail composition
comprises: a first antifungal nail coat composition, which provides
a fungicidal primer coat for relatively quick, substantially
uniform, permeation of terbinafine into, across and onto the nail
plate and the adjacent tissue area, and a second antifungal nail
coat composition which subsequently provides a substantially
uniform film depot coat over the fungicidal primer-coated nail to
act as a nail-protective barrier and a depot for additional
terbinafine which can be gradually released. Thus, the second
antifungal nail composition is applied directly to the
primer-coated nail with no intervening water rinse.
[0046] In a two-coat type embodiment of an antifungal nail coat
composition, the amount of antifungal agent present in the
respective first and second antifungal nail coat compositions can
vary, but preferably, the weight ratio of antifungal agent in the
second nail coat composition relative to that in the first nail
coat composition is less than about one. Depending on the severity
of the infection, the amount of antifungal agent in the first
antifungal nail coat composition can vary in the range of about 0.1
to about 20 percent by weight of the total composition, and the
amount of antifungal agent in the second nail coat composition may
be an amount in the range of about 0.1 to about 15 percent by
weight of the total composition.
[0047] A preferred first antifungal nail coat composition
embodiment for a two-coat type composition is a substantially
clear, colorless solution containing terbinafine at a concentration
in the range of about 0.5 to about 20 weight percent, more
preferably about 10 weight percent dissolved in a volatile,
pharmaceutically acceptable carrier. The volatile carrier
preferably is an alkanol having 2 to about 5 carbon atoms, such as
ethanol, propanol, isopropanol, butanol, isobutanol, and the like.
Ethanol is particularly preferred. The volatile carrier can also
serve as a penetration enhancer.
[0048] A particularly preferred first antifungal nail coat
composition comprises about 10 percent terbinafine in ethanol on a
weight/weight basis. Preferably, the first antifungal nail coat
composition wicks along the capillary system of and across the nail
plate to reach and immobilize fungal spores in the nail plate and
nail bed. A particularly preferred second antifungal nail coat
composition for a two-coat composition embodiment preferably
comprises terbinafine at a concentration in the range of about 0.1
to not more than about 10 weight percent, an effective film-forming
amount of a hydrophilic film-forming polymer and a pharmaceutically
acceptable, volatile carrier as described above as the remainder.
The volatile carrier in the first and second compositions can be
the same or different as desired.
[0049] The hydrophilic polymer may be a film-forming polymer
comprising a vinylpyrrolidone monomer unit, including a
homopolymer, (i.e., polyvinylpyrrolidone), a copolymer and a
complex thereof, a gum, a resin, or the like. The term "copolymer"
as used herein and in the appended claims means any polymer
comprising two or more different monomer repeating units and
includes polymers commonly referred to as "terpolymers,"
"tetrapolymers" and the like.
[0050] Exemplary film-forming polymers containing vinylpyrrolidone
(VP) monomer units, are polyvinylpyrrolidone (PVP), sold in a range
of viscosity grades, and varying weight average molecular weights
in the range of about 8,000 to about 3,000,000 Daltons (PVP K
homopolymer series). PVP is sold under the trade name KOLLIDON.RTM.
CL by BASF Corporation. A USP grade of povidone (PVP) is preferred.
Exemplary film-forming copolymers include
vinylpyrrolidone/vinylaacetate (VA) copolymers available in a range
of mole ratios of VP/VA such as the PVP/VA copolymer series sold by
ISP, and the like. An exemplary VP complex is povidone-iodine
(PVP-I).
[0051] The hydrophilic polymer preferably is a polyvinylpyrrolidone
having a "K" value of about 30 (i.e., a weight average molecular
weight in the range of about 45,000-60,000 Daltons.
[0052] Exemplary gums include agar gum, carrageenan gum, ghati gum,
kara-ya gum, rhamson gum, xanthan gum and the like.
[0053] Exemplary resins include carbomer, a polyacrylic acid
polymer lightly cross-linked with polyalkenyl polyether. It is
commercially available from Noveon Inc. (Cleveland, Ohio) under the
designation "CARBOPOL.RTM.." A particularly preferred grade of
carbomer is that designated as "CARBOPOL.RTM. 940." Other
polyacrylic acid polymers suitable for use are those commercially
available under the designation "PEMULEN.RTM." (Noveon Inc.) and
POLYCARBOPHIL.TM. (A. H. Robbins Company, Inc., Richmond, Va.), is
a polyacrylic acid cross-linked with divinyl glycol. The
PEMULEN.RTM. polymers are copolymers of C.sub.10 to C.sub.30 alkyl
acrylates and one or more monomers of acrylic acid, methacrylic
acid or one of their simple esters cross-linked with an allyl ether
of sucrose or an allyl ether of pentaerythritol.
[0054] There is no limitation on the form (i.e., liquid or powder)
of hydrophilic film-forming polymer used, or the amount used as
long as the nail coat composition can be easily applied to the nail
and form a film thereon.
[0055] The present dual action antifungal nail coat composition can
include one or more substantially non-volatile penetration
enhancers, auxiliary anti-infectives, such as antibacterial agents,
antiseptic agents, and the like, and mixtures thereof. In two-coat
composition embodiments, one or more substantially non-volatile
penetration enhancers can be included in either the first
antifungal nail coat composition or the second antifungal nail coat
composition or in both. The penetration enhancers in the antifungal
nail coat compositions of this invention preferably enhance the
penetration of the drug into the nail as well as the surrounding
skin tissue area.
[0056] Among preferred skin penetration enhancers are ethanol,
propylene glycol, glycerol, ethyl laurate, isopropyl palmitate,
isopropyl myristate, laurocapram (AZONE.RTM.), dioxolanes
(described in U.S. Pat. No. 4,861,764), macrocyclic ketones,
1-decyl-thiolthyl-2-pyrrolidone (HP-101), oxazolidones and
biodegradable penetration enhancers (described in U.S. Pat. Nos.
4,980,378 and 5,082,866 to Wong et al. such as
alkyl-2-(N,N-disubstituted amino) alkanoates (e.g.,
dodecyl-2-(N,N-dimethylamino) isopropionate (DDAIP)),
N,N-disubstituted amino alkanol alkanoates) and mixtures thereof.
Aliphatic and aromatic alcohols are primarily nail penetration
enhancers.
[0057] The penetration enhancer is present in an amount sufficient
to enhance the penetration of the antifungal agent. The specific
amount varies necessarily according to the desired release rate and
the specific antifungal agent used. Generally, the penetration
enhancer is present in an amount ranging from about 0.1 weight
percent to about 25 weight percent, based on the total weight of
the antifungal nail coat composition. Preferably, the penetration
enhancer is present in an amount ranging from about 0.1 weight
percent to about 10 weight percent, more preferably, in an amount
ranging from about 0.5 weight percent to about 5 weight percent of
the antifungal nail coat composition.
[0058] In general, suitable penetration enhancers can be chosen
from those listed above, as well as aliphatic and aromatic
alcohols, sulfoxides, fatty acids, fatty acid esters, polyols,
amides, surfactants, terpenes, alkanones, organic acids and
mixtures thereof. See generally Chattaraj, S.C. and Walker, R. B.,
Penetration Enhancer Classification, pp.5-20 in Maibach, H. I., and
Smith, H. E., (eds.), Percutaneous Penetration Enhancers, CRC
Press, Inc., Boca Raton, Fla. (1995) and Buyuktimkin, N., et al.,
Chemical Means of Transdermal Drug Permeation Enhancement, in
Ghosh, T. K., and Pfister, W. R. (eds.) Transdermal and Topical
Drug Delivery Systems, Interpharm Press, Inc., Buffalo Grove, Ill.
(1997).
[0059] Suitable alcohols include, without limitation, ethanol,
propanol, butanol, pentanol, hexanol, octanol, nonanol, decanol,
2-butanol, 2-pentanol, benzyl alcohol, phenoxyethanol, caprylic
alcohol, decyl alcohol, lauryl alcohol, 2-lauryl alcohol, myristyl
alcohol, cetyl alcohol, stearyl alcohol, oleyl alcohol, linolyl
alcohol, linolenyl alcohol and mixtures thereof. Volatile aliphatic
alcohols having 2 to about 5 carbon atoms can provide a dual
function of serving both as volatile carrier and penetration
enhancer. The aromatic alcohols, such as benzyl alcohol,
phenoxyethanol, and the like can provide a dual function of serving
both as a substantially non-volatile, permeation enhancer and
auxiliary anti-infective. Preferred alcohols are ethanol and benzyl
alcohol.
[0060] Suitable sulfoxides include dimethylsulfoxide (DMSO),
decylmethylsulfoxide, and mixtures thereof.
[0061] Suitable fatty acids include valeric, heptanoic, pelargonic,
caproic, capric, lauric, myristic, stearic, oleic, linoleic,
linolenic, caprylic, isovaleric, neopentanoic, neoheptanoic,
neononanoic, trimethyl hexanoic, neodecanoic and isostearic acids,
and mixtures thereof.
[0062] Suitable fatty acid esters include isopropyl n-butyrate,
isopropyl n-hexanoate, isopropyl n-decanoate, isopropyl myristate,
isopropyl palmitate, octyldodecyl myristate, ethyl acetate, butyl
acetate, methyl acetate, methylvalerate, methylpropionate, diethyl
sebacate, ethyl oleate, ethyl laurate and mixtures thereof.
Suitable polyols include propylene glycol, polyethylene glycol,
ethylene glycol, diethylene glycol, triethylene glycol, dipropylene
glycol, glycerol, propanediol, sorbitol, dextrans, butanediol,
pentanediol, hexanetriol, and mixtures thereof.
[0063] Suitable amides include urea, dimethylacetamide,
diethyltoluamide, dimethylformamide, dimethyloctamide,
dimethyldecamide, pyrrolidone derivatives,
1-alkyl-4-imidazolin-2-one, cyclic amides, hexamethylenelauramide
and its derivatives, diethanolamine, triethanolamine and mixtures
thereof. Suitable pyrrolidone derivatives include
1-methyl-2-pyrrolidone, 2-pyrrolidone, 1-lauryl-2-pyrrolidone,
1-lauryl-4-carboxy-2-pyrrolidone, 1-methyl-4-carboxy-2-pyrrolidone,
1-hexyl-4-carboxy-2-pyrrolidone, 1-decylthioethyl-2-pyrrolidone
(HP-101), N-cyclohexylpyrrolidone,
1-methyl-4-methoxycarbonyl-2-pyrrolidone,
1-hexyl-4-methoxycarbonyl-2-pyrrolidone,
1-lauryl-4-methoxycarbonyl-2-pyrrolidone,
N-dimethylaminopropylpyrrolidone, N-cocoylpyrrolidone,
N-tallowylpyrrolidone, fatty acid esters of
N-(2-hydroxymethyl)-2-pyrrolidone, and mixtures thereof. Suitable
cyclic amides include, 1-dodecylazacycloheptan-2-one (laurocapram,
AZONE.RTM.), 1-geranylazacycloheptan-2-one,
1-farnesylazacycloheptan-2-one,
1-geranylgeranylazacycloheptan-2-one,
1-(3,7-dimethyloctyl)azacycloheptan-2-one, 1-(3
,7,11-trimethyloctyl)azacycloheptan-2-one,
1-geranylazacyclohexan-2-one, 1-geranylazacyclopentan-2,5-dione,
1-farnesylazacyclopentan-2-one, and mixtures thereof.
[0064] Suitable surfactants include anionic surfactants, cationic
surfactants, nonionic surfactants, amphoteric surfactants, bile
salts and lecithin. Suitable anionic surfactants include sodium
laurate, sodium lauryl sulfate, and mixtures thereof. Suitable
cationic surfactants include cetyltrimethylammonium bromide,
tetradecyltrimethylammonium bromide, benzalkonium chloride,
octadecyltrimethylammonium chloride, cetylpyridinium chloride,
dodecyltrimethylammonium chloride, hexadecyltrimethylammonium
chloride, and mixtures thereof. Suitable nonionic surfactants
include
.alpha.-hydro-.omega.-hydroxypoly(oxyethylene)-poly(oxypropyl)
poly(oxyethylene) block copolymers, polyoxyethylene ethers,
polyoxyethylene sorbitan esters, polyethylene glycol esters of
fatty alcohols, and mixtures thereof. Suitable
.alpha.-hydro-.omega.-hydroxy-poly(oxyethylene)-poly(oxypropyl)
poly(oxyethylene) block copolymers include Poloxamers 182, 184,
231, and mixtures thereof. Suitable polyoxyethylene ethers include
PEG-4 lauryl ether (BRIJ.RTM. 30), PEG-2 oleyl ether (BRIJ.RTM.
93), PEG-10 oleyl ether (BRIJ.RTM. 96), PEG-20 oleyl ether
(BRIJ.RTM. 99), and mixtures thereof. Suitable polyoxyethylene
sorbitan esters include the monolaurate (TWEEN.RTM. 20) the
monopalmitate (TWEEN.RTM. 40), the monostearate (TWEEN.RTM. 60),
the monooleate (TWEEN.RTM. 80), and mixtures thereof. Suitable
polyethylene glycol esters of fatty acids include polyoxyethylene
(8) monostearate (MYRJ.RTM. 45), polyoxyethylene (30) monostearate
(MYRJ.RTM. 51), the polyoxyethylene (40) monostearate (MYRJ.RTM.
52), and mixtures thereof.
[0065] Suitable amphoteric surfactants include, without limitation
thereto, lauramidopropyl betaine, cocamidopropyl betaine, lauryl
betaine, cocobetaine, cocamidopropylhydroxysultaine, aminopropyl
laurylglutamide, sodium cocoamphoacetate, sodium lauroamphoacetate,
disodium lauroamphodiacetate, disodium cocoamphodiacetate, sodium
cocoamphopropionate, disodium lauroamphodipropionate, disodium
cocoamphodipropionate, sodium lauriminodipropionate, disodium
cocoamphocarboxymethylhydroxypropylsulfate, and the like.
[0066] Suitable bile salts include sodium cholate, sodium salts of
laurocholic, glycolic and desoxycholic acids, and mixtures
thereof.
[0067] Suitable terpenes include D-limonene, .alpha.-pinene,
.beta.-enrene, .alpha.-terpineol, terpinen-4-ol, carvol, carvone,
pulegone, piperitone, menthone, menthol, geraniol, cyclohexene
oxide, limonene oxide, .alpha.-pinene oxide, cyclopentene oxide,
1,8-cineole, ylang ylang oil, anise oil, chenopodium oil,
eucalyptus oil, and mixtures thereof. Suitable alkanones include
N-heptane, N-octane, N-nonane, N-decane, N-undecane, N-dodecane,
N-tridecane, N-tetradecane, N-hexadecane, and mixtures thereof.
Suitable organic acids include citric acid, succinic acid,
salicylic acid, salicylates (including the methyl, ethyl and propyl
glycol derivatives), tartaric acid, and mixtures thereof.
[0068] A preferred, substantially non-volatile, penetration
enhancer comprises an N,N-di(C.sub.1-C.sub.8) alkylamino
substituted, (C.sub.4-C.sub.18) alkyl (C.sub.2-C.sub.18) carboxylic
ester or pharmaceutically acceptable acid addition salt thereof. As
used herein, the term "(C.sub.4-C.sub.18) alkyl (C.sub.2-C.sub.18)
carboxylic ester" means an ester of a (C.sub.4-C.sub.18) alcohol
and a (C.sub.2-C.sub.18) carboxylic acid. The term
"N,N-di(C.sub.1-C.sub.8) alkylamino substituted," in reference to a
(C.sub.4-C.sub.18) alkyl (C.sub.2-C.sub.18) carboxylic ester means
that either the alcohol portion or the carboxylic acid portion from
which the ester is prepared bears an amino substituent
NR.sub.xR.sub.y, wherein R.sub.x and R.sub.y are each independently
a (C.sub.1-C.sub.8) alkyl group. Preferably R.sub.x and R.sub.y are
both methyl groups.
[0069] Preferred are dodecyl-2-(N,N-dimethylamino) propionate
(DDAIP); dodecyl-2-(N,N-dimethylamino)-acetate (DDAA);
1-(N,N-dimethylamino)-2-propyl dodecanoate (DAIPD);
1-(N,N-dimethylamino)-2-propyl myristate (DAIPM);
1-(N,N-dimethylamino)-2-propyl oleate (DAIPO); and pharmaceutically
acceptable acid addition salts thereof.
[0070] A particularly preferred skin permeation enhancer is DDAIP,
alone or in combination with an auxiliary permeation enhancer.
DDAIP.HCl is available from Steroids, Ltd. (Chicago, Ill.) and
Pisgah Laboratories (Pisgah Forest, N.C.). Particularly preferred
is the hydrochloride of DDAIP (DDAIP.HCl). The preparation of DDAIP
and crystalline acid addition salts thereof is described in U.S.
Pat. No. 6,118,020 to Buyuktimkin, et al., which is incorporated
herein by reference. Long chain similar amino substituted, alkyl
carboxylic esters can be synthesized from readily available
compounds as described in U.S. Pat. No. 4,980,378 to Wong, et al.,
which is incorporated herein by reference to the extent that it is
not inconsistent herewith.
[0071] The term "anti-infective agent" as used herein includes a
topical antibacterial, antiseptic, or the like, that can augment
the efficacy of the dual action antifungal nail coat composition.
Suitable antibacterial agents include bacteriostatic preservatives,
such as benzyl alcohol, phenoxyethanol, phenethylalcohol,
iodopropynl butyl carbamate, paraben, and the like. Benzyl alcohol
is particularly preferred, and when present can serve a dual
purpose as penetration enhancer and anti-infective.
[0072] Suitable antiseptic agents include alcohol (i.e., ethanol,
isopropanol), halogen containing compounds, (i.e., povidone-I,
triclosan, and the like); quaternary ammonium compounds (i.e.,
benzethonium chloride, cetylpyridimum chloride, and the like).
[0073] Those skilled in the art will recognize that one or more of
the foregoing ingredients can serve more than one function.
[0074] A preferred dual action, one-coat type antifungal nail coat
composition embodiment comprises: [0075] an effective fungicidal
amount of an antifungal agent; [0076] a permeation enhancing amount
of a substantially non-volatile, [0077] permeation enhancer
selected from the group consisting of an N,N-di(C.sub.1-C.sub.8)
alkylamino substituted, (C.sub.4-C.sub.18) alkyl (C.sub.2-C.sub.18)
carboxylic ester or pharmaceutically acceptable acid addition salt
thereof, a pharmaceutically acceptable alcohol, and mixtures
thereof; [0078] a film-forming amount of a hydrophilic polymer; and
[0079] a pharmaceutically acceptable, volatile organic carrier.
[0080] Preferably, the one-coat antifungal nail coat composition is
a substantially clear formulation.
[0081] A preferred dual action, one-coat type embodiment of
antifungal nail coat composition comprises on a total composition
weight basis: [0082] antifungal agent in an amount in the range of
about 0.1 to about 20 weight percent, more preferably in the range
of about 0.5 to about 15 weight percent; most preferably in the
range of about 1 to about 5 weight percent; [0083] a substantially
non-volatile permeation enhancer in a total amount in the range of
about 0.1 to about 25 weight percent, more preferably in the range
of about 1 to about 10 weight percent; [0084] a hydrophilic
film-forming polymer in an amount in the range of about 0.1 to
about 5 weight percent, more preferably in the range of about 0.25
to about 1 weight percent; and [0085] the remainder comprising a
pharmaceutically acceptable volatile organic carrier. A preferred
volatile organic carrier is an aliphatic alcohol preferably present
in an amount in the range of about 50 to about 99.5 weight percent,
more preferably in the range of about 85 to about 99, based on a
total composition weight basis.
[0086] A particularly preferred substantially clear, dual-action,
one-coat type antifungal nail coat composition comprises on a total
composition weight basis: terbinafine hydrochloride present in an
amount in the range of about 0.5 to about 10 weight percent, more
preferably in the range of about 1 to about 5 weight percent;
[0087] DDAIP.HCl present in an amount in the range of about 0.1 to
about 25 weight percent, more preferably in the range of about 0.1
to about 10 weight percent; [0088] benzyl alcohol present in an
amount in the range of about 0.1 to about 10 weight percent, more
preferably in the range of about 0.5 to about 1.5 weight percent;
[0089] polyvinylpyrrolidone present in an amount in the range of
about 0.1 to about 5 weight percent, more preferably in the range
of about 0.25 to about 1 weight percent; and [0090] the remainder
being ethanol.
[0091] In a two-coat type embodiment utilizing first and second
antifungal nail coat compositions, the second antifungal nail coat
composition preferably is formulated so that the film coat
deposited on fingernails is substantially more resistant to ready
removal with water than the primer film coat deposited on
toenails.
[0092] A particularly preferred, two-coat type dual action
antifungal nail coat composition comprises, in the first or primer
antifungal nail coat composition, on a total composition weight
basis, about 10 weight percent terbinafine in ethanol, and in the
second antifungal nail coat composition preferably not more than
about 5 weight percent terbinafine. A presently preferred second or
depot antifungal nail coat composition comprises about 20 parts by
weight polyvinylpyrrolidone, about 3 parts by weight terbinafine,
and about 47 parts by weight ethanol.
[0093] Based on in vitro test studies using human nail clippings,
it was found that terbinafine applied as a 10% solution in ethanol
can diffuse across a nail membrane and, in a period of about one
hour, can reach a concentration above the minimum inhibition
concentration (MIC) for fungi.
[0094] Fungal infection of a toenail or fingernail may be
ameliorated or prevented by a one-coat method, or a two-coat method
as described below.
[0095] A one-coat type dual action antifungal nail coat composition
can be applied to provide a substantially uniform fungicidal
coating on a fungally susceptible or infected nail and adjacent
skin tissue and maintained in contact therewith for a period of at
least about 0.5 hour. In a one-coat method, the nail coat
composition can be removed subsequently by rinsing with water. In a
multiple-coat method, the composition can be re-applied at least
twice with or without an intervening water rinse. The nail coat
composition is preferably applied in a daily regimen for a period
sufficient to achieve fungicidal efficacy.
[0096] A two-coat type dual action antifungal nail coat composition
of this invention can be applied by the following multiple-coat
method.
[0097] (1) A first antifungal nail coat composition containing an
effective fungicidal amount of antifungal agent is applied at least
once to an infected fingernail or toenail, and surrounding skin
area, to provide an active fungicidal primer coat;
[0098] (2) The active fungicidal primer coat is allowed to
substantially dry for about 10 minutes or until the fungicidal
primer-coated nail is substantially dry to the touch; and then
[0099] (3) The substantially dry fungicidal primer-coated nail is
coated with a sufficient fungicidal amount of a second antifungal
nail coat composition to provide a fungicidal film coat thereon for
further release of antifungal agent to the nail.
[0100] In the initial period of a fungicidal regimen with a
two-coat type dual action antifungal nail coat composition,
multiple applications of the first antifungal nail coat composition
can be applied, by performing sequential steps (1) and (2) at least
twice before performing step (3) to further optimize the
bioavailability of antifungal agent.
[0101] The methods of this invention are preferably practiced daily
until new nail growth is visibly free of fungal infection.
[0102] It was found that the practice of a two-coat method of this
invention with terbinafine extended the residence time of the
terbinafine applied from the first antifungal nail coat
composition, that the hydrophilic polymer film coat of the second
antifungal nail coat composition promoted the formation of an
internal and external barrier membrane, and that a high efficacy in
ameliorating or preventing onychomycosis within a relatively short
period of about four weeks was achieved.
[0103] The nail coat compositions of the present invention can be
applied to the nail by any convenient method, such as by brushing
or spraying. Preferably the applied composition is substantially
dry to the touch within a period in the range of about 0.5 to about
10 minutes, more preferably within a period in the range of about
one to about five minutes, depending upon the amount of volatile
organic carrier present.
[0104] The fungicidal nail coat compositions may be provided in kit
form with instructional indicia included therein for use. The first
and second antifungal coat compositions of a two-coat dual action,
antifungal nail coat composition may be individually packaged in
similar or dissimilar shaped packages or are color coded to visibly
distinguish the first and second compositions from one another to
aid the user in following the therapeutical order of
application.
[0105] Instructional indicia includes, without limitation, printed
media, aural media, visual aids, electronic media or a combination
thereof which inform and instruct the user. Printed media includes,
but is not limited to, labels, pamphlets, books, flyers and the
like. Aural media includes, but is not limited to, tape recordings,
audio compact disks, records, and the like. Visual aids include,
but are not limited, to photographs, slides, movies, videos, DVDs,
and the like. Electronic media includes all forms of electronic
data storage media, such as, but not limited to, diskettes,
interactive CD-ROMs, interactive DVDs, and the like.
[0106] The following examples are intended to illustrate, but not
limit, the present invention.
EXAMPLE 1
[0107] The preliminary efficacy and safety of terbinafine
hydrochloride in a two-coat type dual action, antifungal nail coat
composition and method of this invention was studied with patients
having toenail and/or fingernail fungal infection. The patients
participated in an open label, single hospital pilot clinical study
over a period of three months.
[0108] Up to 20 patients were selected assessed as having mild to
severe onychomycosis, as measured by using a scale of infection,
based on nail plate separation from the nail bed, hyperkeratosis,
and discoloration. The extent of onychomycosis, hyperkeratosis, and
discoloration were assessed using the following scale ratings:
[0109] Onychomycosis [0110] 0=absence of separation of nail plate
from nail bed. [0111] 1=.ltoreq.50% separation of nail plate.
[0112] 2=>50% but s 75% separation of nail plate. [0113]
3=>75% separation of nail plate. [0114] Hyperkeratosis [0115]
0=absence of subungual debris. [0116] 2=thickening of .ltoreq.50%
of the subungual region. [0117] 2=>505 but .ltoreq.75%
thickening of the subungual region. [0118] 3=>75% thickening of
the subungual region. [0119] Discoloration [0120] 0=absence of any
unusual coloration (white, yellow, etc.) of the nail plate. [0121]
1=discoloration extending to .ltoreq.50% of the nail plate. [0122]
2=discoloration extending to >50% but 5.75% of the nail plate.
[0123] 3=discoloration extending to >75% of the nail plate.
[0124] For inclusion in the study, the criteria were: onychomycosis
patients between the ages of 18-70 years, having a nail involvement
of at least 25% of the whole nail surface that included any
destroyed or missing part of the nail plate. Onychomycosis of the
finger nail or toenail was confirmed as follows by KOH staining
microscopic examination and fungal culture.
[0125] The nail plate and hard debris were softened by leaving the
fragments, along with several drops of potassium hydroxide (25% KOH
with 5% glycerine), in a watch glass covered with a petri dish for
24 hours. Light microscope was used for the fungal examination. The
small fragments of scale were placed on a microscope slide and a
coverslip was applied. The preparation was studied carefully at low
power. Dermatophytes appear as translucent branching, rod-shaped
filaments of uniform width. If the presence of hyphae is confirmed
by examination with the 40.times. objective, the test result is
judged as positive.
[0126] Fungal culture was carried out using the standard culture
medium, Sabouraud's agar, (agar 18 g, peptone 10 g, glucose 40 g,
distilled water 1000 ml). Most medically important fungi are grown
aerobically on this culture medium over an incubation period of
about 24 hours to about 48 hours at a temperature of about
28.degree. C.
[0127] The criteria for exclusion from the study were:
onychomycosis caused by molds (Candida sp.); hypersensitivity to
terbinafine; abnormal liver function (twice the upper limit value);
receipt of topical treatment within 2 weeks or oral treatment
within two months; concurrent treatment with H-blockers, antacid,
rifampin, phenobarbital, phenytoin, carbamazepine, terfenadine
(e.g. SELDANE.TM.) or digoxin; use of any investigational drugs
with one month; psoriasis or history of psoriasis; serious
concurrent disease that might influence the trial; and pregnant
women or nursing mothers.
[0128] Twenty patients (six females, 14 males) between the ages of
35-59 years, with an average age of 46 years, met the inclusion
criteria. Of these 20 subjects, 17 completed 12 weeks of treatment.
At the start of the study, the extent of onychomycosis was assessed
as mild (i.e., .ltoreq.40% infected nail) for 15%, and as severe
(i.e., >40% infected nail) for the remaining 85% of the 20
patients. Of the 20 patients, 45% of the patients had separation of
nail plate; 45% had hyperkeratosis; and 10% had discoloration.
[0129] The primary efficacy criteria were mycological cure based on
achieving a negative KOH staining microscopic examination and a
negative fungal culture.
[0130] The secondary efficacy criteria were the physicians's
assessment of the mycological cure and clinical efficacy. Clinical
efficacy evaluation was assessed as follows: "Cleared" (i.e., no
signs of mycosis, without residual nail deformity, no requirement
for further therapy); "Markedly Improved" (i.e., minimal nail
involvement with significantly decreased signs of mycosis; and
"Slightly to Moderately Improved" (i.e., slight to moderate
reduction in extent of nail involvement and signs of mycosis).
[0131] After the completion of the study, the clinical safety and
efficacy of administration were analyzed by investigators based on
adverse events, KOH staining microscopic examination, fungal
culture, clinical efficacy assessment (i.e., planimetric
measurement of the involved area, photographic comparison of new
nail growth, and reduction in extent of nail involvement) and the
physician's global evaluation.
[0132] The primary safety parameters included adverse events, vital
signs, clinical laboratory tests, physical examinations, and
electrocardiograms (ECG).
[0133] The patients assigned to the study were each provided with
two bottles having brush applicators, each bottle containing nail
coat composition (about 20 grams in each bottle), and identified as
"A" and "B". Bottle "A" contained terbinafine hydrochloride 10%
(weight/weight) in ethanol. Bottle "B" contained 20 parts by weight
polyvinylpyrrolidone (PVP, KOLLIDON.RTM. 30, weight average
molecular weight in the range of about 45,000-60,000 Daltons), 3
parts by weight terbinafine hydrochloride, and 47 parts by weight
ethanol.
[0134] The patients were instructed to cleanse their feet or hands
by using warm water, and cut or clean infected nails as much as
possible, but not to file the nails. The patients were also
instructed to apply the antifungal nail coat composition on the
infected nail directly once each night substantially immediately
after washing their feet.
[0135] The patients were instructed to first apply Solution A with
the brush, let Solution A dry, and then apply Solution B with the
brush and let Solution B dry. There were no limitations to avoid
wetting or washing their feet. The coating was easy to wash off
before re-applying the dual action antifungal nail coat
composition. The patients were instructed that, after washing off
the coating, the patient re-apply the antifungal nail coat
composition right away. The doctors encouraged the patients to use
the antifungal nail coat composition on a daily basis, especially
for the first month.
[0136] The efficacy, based on primary efficacy (mycological cure),
clinical efficacy (appearance of the new nail, disappearance of
signs and symptoms), and total efficacy (i.e., both mycological
evaluation and clinical evaluation assessments) at the end of the
first, second and third month of the study period is summarized in
Table 1.
TABLE-US-00001 TABLE 1 Month 1 Month 2 Month 3 Patients % Patients
% Patients % Evaluated 18 100 17 100 17 100 Primary Efficacy 7 38.9
8 47.1 9 52.9 Clinical Efficacy 6 33.3 10 58.8 16 94.1 Total
Efficacy 7 38.9 8 47.1 9 52.9
[0137] As shown in Table 1, based on the assessed change in nail
involvement, change in signs of fungal infection, and new nail
growth, the clinical efficacy (including patients rated as
"slightly to moderately improved," "markedly improved," and
"cleared") at the end of the first, second, and third month of
treatment, was 33.3%, 58.8% and 94.1%, respectively.
[0138] As shown in Table 2, the number of patients initially
assessed as having severe onychomycosis decreased at the end of the
first, second, and third month of the study period, and
concurrently, the number of patients assessed as having mild
onychomycosis increased.
TABLE-US-00002 TABLE 2 Month 0 Month 1 Month 2 Month 3
Onychomycosis Patients % Patients % Patients % Patients % Evaluated
20 100 18 100 17 100 17 100 Mild (.ltoreq.40%) 3 15 4 22.2 4 23.5 6
35.3 Severe (.gtoreq.40%) 17 85 14 77.8 13 76.5 11 64.7
[0139] One patient having "mild" onychomycosis and one patient
having "severe" onychomycosis were judged as showing significant
improvement at the end of the third month.
[0140] During the study period, the patients also maintained a
diary from which the patient's experiences of any adverse events
were recorded. No adverse events were reported by any of the
patients during the study period.
[0141] It is recognized that new nail growth takes time. The nail
reportedly grows continuously at the rate of 3-4 millimeters (mm) a
month (0.112 to 0.132 mm a day), so some 4.5-5 months are required
for a complete renewal of the nail. It is also recognized that the
speed of nail growth differs between individuals as well as age
groups (nail growth being more rapid in the young), and that
certain health disorders and medications can upset the rate of
growth. Thus, mycological evaluation was judged as the most proper
objective primary efficacy criteria to best predict full future
clinical efficacy. The efficacy of the two-coat type dual action
antifungal nail composition within the short-term study period as
judged safe and effective for ameliorating onychomycosis of varying
intensity.
EXAMPLE 2
[0142] The uptake of antifungal agent by a nail substrate was
evaluated in vitro using human nail clippings collected from one
individual. The nail clippings were cleaned and extracted with
anhydrous ethyl alcohol for several days before applying the
antifungal agent, terbinafine hydrochloride.
[0143] About 15 mL of four antifungal containing solutions each
were prepared comprising the following indicated amount, on a total
composition volume basis, terbinafine hydrochloride, volatile
organic carrier (ethanol), film-forming hydrophilic polymer
(polyvinylpyrrolidone (PVP)), or penetration enhancer
dodecyl-2-(N,N-dimethylamino) isopropionate hydrochloride
(DDAIP.HCl)).
[0144] Solution A. 10 weight percent terbinafine hydrochloride in
anhydrous ethyl alcohol.
[0145] Solution B. 10 weight percent terbinafine hydrochloride and
10 weight percent PVP (KOLLIDON.RTM. 30, BASF) in anhydrous ethyl
alcohol.
[0146] Solution C. 10 weight percent terbinafine hydrochloride and
0.5 weight percent DDAIP.HCl in anhydrous ethyl alcohol.
[0147] Solution D. 10 weight percent terbinafine hydrochloride and
1 weight percent DDAIP.HCl in anhydrous ethyl alcohol.
[0148] The nail clippings were separately immersed in about 5 mL of
each of solution A, B, C, and D (solid:liquid ratio of about 1:10),
and the uptake of terbinafine was determined by measuring the
concentration of terbinafine in the solution as a function of time
over a period starting from immersion to about 24 hours.
Measurement was made using High Performance Liquid Chromatography
(HPLC) technique using a Waters Alliance HPLC. (Waters Symmetry
C18, 3.5 nm 4.2.times.75 mm column was equipped for the
separations, UV 224 nm for detection, flow rate 1.5 mL/min.,
injection 20 .mu.L). The buffer was composed of two parts
triethylamine and 1000 parts of deionized water and the pH was
adjusted to pH 7 with phosphoric acid. The mobile phase composition
was 25 parts of buffer and 75 parts acetonitrile.
[0149] As shown graphically in FIG. 1, an initially fast decrease
in the solution concentration of terbinafine was observed in all
cases, which gradually approached equilibrium after about five
hours, remaining substantially unchanged up to 24 hours, indicating
that saturation had been reached. Uptake from Solution A reached
equilibrium in less than about one hour, somewhat sooner than from
Solutions B, C or D. In all cases, the average amount of
terbinafine uptake was judged to be about 5.2 mg/100 mg nail or
about 5.2% on a nail weight basis.
[0150] The terbinafine-treated nail clippings were then separately
recovered from each test solution and rinsed with 10 mL of ethyl
alcohol to remove antifungal liquid from the surface cavity. The
rinsed nail clippings from each test were then separately immersed
in another 5 mL portion of anhydrous ethyl alcohol to assess the
rate of terbinafine release from the nail structure, by determining
the concentration of terbinafine hydrochloride released as a
function of time using the HPLC technique described above. The
amount of terbinafine hydrochloride initially released from the
nail, based on release measurements over a period of about 48
hours, was greater from nail treated with Solution A, than from
nail treated with Solution B, C, or D. As shown graphically in FIG.
2, the amount of terbinafine hydrochloride retained in the nail
reached equilibrium in a period of about 10 hours. The order of
efficacy of treatment was Solution D>Solution C>Solution
B>Solution A.
[0151] FIG. 3 graphically shows the retention of terbinafine
hydrochloride update in the nails treated with Solutions C and D.
The data indicated that the film-forming polymer in Solution B, and
the penetration enhancer in Solutions C and D, contributed
beneficially to increasing the residence time of terbinafine in the
nail.
EXAMPLE 3
[0152] The permeation of terbinafine hydrochloride by human nail
clippings as a function of time was compared using Solution A and
C, prepared as in Example 2. Nail clippings having a substantially
similar dry thickness (+/-5%) were selected. A selected nail
clipping was anchored by being placed between two open metal
frames, a sealant material was placed between the rim of the frame
and the edge of the nail, and the edges of the nail were then
compressed to stabilize the nail and provide a nail holder. The
nail holder thus had an opening for permeation and was sealed
against leakage when the anchored nail was placed in a horizontal
Franz diffusion cell as a permeable membrane. The volume capacity
of each of the donor cell and receiving cell was 3 mL, and the
permeation area of about 78.5 square mm. The donor solution was the
antifungal solution (Solution A or Solution C) and the receiver
solution was anhydrous ethyl alcohol. The receiver solution was
sampled periodically over a period of up to about 100 hours, and
analyzed by HPLC, as in Example 2.
[0153] The cumulative permeation of terbinafine hydrochloride in
the receiver is graphically shown in FIG. 4, and indicates an
enhanced permeation of terbinafine hydrochloride through the nail
from Solution C containing 10% terbinafine hydrochloride and 0.5%
DDAIP.HCl over that of Solution A containing 10% terbinafine
hydrochloride in anhydrous ethyl alcohol.
EXAMPLE 4
[0154] This example illustrates formulations for one-coat type dual
action, antifungal nail coat compositions, (A), (B), (C), (D) and
(E).
TABLE-US-00003 TABLE 3 WEIGHT PERCENT INGREDIENT (A) (B) (C) (D)
(E) Terbinafine.cndot.HCl 1 5 10 1 1 DDAIP.cndot.HCl 0.5 0.5 0.5
2.5 5 PVP, USP 0.5 0.5 0.5 0.5 0.5 Benzyl alcohol 0.75 0.75 0.75
0.75 0.75 Ethanol to 100% q.s. q.s. q.s. q.s. q.s. q.s. = quantity
sufficient
EXAMPLE 5
[0155] This example illustrates, in a recognized guinea pig model
of dermatophytosis caused by Trichophyton mentagrophytes (T.
mentagrophytes) (ATCC 24953), the in vivo clinical and fungicidal
efficacy of one-coat type dual action, antifungal nail coat
compositions containing varying amounts of terbinafine
hydrochloride and of penetration enhancer, DDAIP.HCl. Ten
compositions were prepared having the amounts indicated in Table
4.
TABLE-US-00004 TABLE 4 Weight Percent Ingredient Terbinafine PVP,
Benzyl Ethanol Example HCl DDAIP.cndot.HCl USP Alcohol to 100% 5(A)
None None 0.5 0.75 q.s. (control) 5(B) None 0.5 0.5 0.75 q.s. 5(C)
1 None 0.5 0.75 q.s. 5(D) 5 None 0.5 0.75 q.s. 5(E) 10 None 0.5
0.75 q.s. 5(F) 1 0.5 0.5 0.75 q.s. 5(G) 1 2.5 0.5 0.75 q.s. 5(H) 1
5 0.5 0.75 q.s. 5(I) 5 0.5 0.5 0.75 q.s. 5(J) 10 0.5 0.5 0.75
q.s.
[0156] The procedures of the in vivo evaluation protocol used were
in compliance with the Animal Welfare Act, the Guide for the Care
and Use of Laboratory Animals, and the Office of Laboratory Animal
Welfare. The protocol also was approved by the Institutional Animal
Care and Use Committee (IACUC), and the IACUC Guidelines were
followed. The evaluation was carried out at the Center for Medical
Mycology and Mycology Reference Laboratory of Case Western Reserve
University, Cleveland, Ohio.
[0157] Male albino Guinea-Pigs Harlan-Sprague-Dawley (San Diego,
Calif.) having a body weight of about 400 to about 450 grams were
acclimated for a minimum of five days prior to use. The
environmental controls for the animal room were set to maintain a
temperature in the range of about 16 to about 22.degree. C., a
relative humidity in the range of about 30 to about 70%, and a 12
hour light/12 hour dark cycle. Guinea pigs are naturally
susceptible to dermatophyte infection and need no special
manipulation, such as immunosuppression.
[0158] Each test guinea pig was anesthetized with an intramuscular
(IM) injection of 0.1 ml of an anesthetic cocktail of xylazine,
ketamine and acepromazine (3:3:1 by volume). Using an electric
shaver, hair was clipped on the left side of the guinea pig's back.
A closer shave was given with a safety razor. Using a stencil, a
shaved skin area of about 2.5.times.2.5 cm.sup.2 square was marked
in quadrants, and the marked skin area was abraded with sterile
fine grit sandpaper. The guinea pig was then infected topically by
thoroughly rubbing onto the abraded skin a cell suspension of T.
mentagrophytes, (ATCC 24953).
[0159] The T. mentagrophytes suspension was prepared by
sub-culturing T. mentagrophytes (from frozen stock) on Potato
Dextrose Agar (PDA) (Difco Laboratories) plates and incubating the
plates at a temperature of about 30.degree. C. for a period of
about five to about seven days. The colonies were scraped from the
plate using sterile saline solution (NaCl 0.85%). After washing
three times with sterile saline solution, the conidia were
re-suspended in sterile saline solution. A ten-fold dilution of
conidia suspension was prepared and counted using a hemacytometer.
A working suspension of conidia was prepared at a final
concentration of 1.times.10.sup.7 Colony Forming Units (CFU) per
100 microliters normal saline solution. The inoculum counts of the
ten-fold dilution of T. mentagrophytes working conidial suspension
was checked by plating the suspension onto Sabouraud Dextrose Agar
(Difco Laboratories) media, incubating the plate at a temperature
of about 30.degree. C. for a period of about three to about four
days, and then determining the colony counts.
[0160] Three days after the inoculation and infection with the
dermatophyte, the guinea pigs were each treated, once daily for a
period of seven days, with 0.1 mL/application of one of the
selected nail coat compositions, 5(A-J), listed in Table 4. Three
days after completion of the seven-day test period, mycological and
clinical efficacy was examined.
[0161] Mycological efficacy was examined by removing hair samples
with a sterile forceps from four quadrants, (10 representative
hairs per quadrant). The hair samples were planted in a
corresponding quadrant on a Potato Dextrose Agar plate and
incubated at a temperature of about 30.degree. C. for about two
days. Following the two-day incubation period, the fungal growth at
the hair root was examined under a stereo-microscope. The
effectiveness of a test composition in reducing the number of
mycologically positive hair samples per treated animal group was
expressed as percentage efficacy relative to the untreated control
group of animals using the following formula: %
efficacy=100-(T.times.100/K), where T=positive hair in the test
group and K=positive hair in the untreated control group.
[0162] Four guinea pigs were tested with the composition of Ex.
5(A) as a placebo (vehicle control) group (Group 1), five guinea
pigs were tested with each one of the example formulations (Exs.
5(B-J) shown in Table 4, (identified as Groups 2-10 respectively),
and one group of four guinea pigs was maintained as an infected
control group (Group 11).
[0163] The hairs from the infected, control guinea pigs (Group 11)
showed growth of fungal filaments indicating invasion of the hair
roots. Substantially similar invasion of the hair roots was noted
in the infected guinea pigs treated with placebo (Group 1) and with
the drug-free composition of Ex. 5(B) (Group 2). All of the
compositions containing terbinafine HCl, Exs. 5(C-J), had
mycological efficacy as demonstrated by the absence of fungal
elements in the hair.
[0164] Clinical efficacy was assessed by examining local changes in
the appearance of the skin and regrowth of hair at the test sites,
using the following numerical score criteria: 0=no lesions; 1=few
slightly erythematous places on the skin; 2=well defined redness,
swelling with bristling hairs; 3=large areas of marked redness
incrustation, scaling, bald patches, ulcerated in places; 4=partial
damage to the integument and loss of hair; and 5=extensive damage
to the integument and complete loss of hair at the site of
infection. The assessment of clinical evaluation in the change of
scores per treated animal group was expressed as a percentage
relative to the untreated control group of animals using the
following formula: % efficacy=100-(T.times.100/K), where T=scores
in the test group and K=scores in the untreated control group.
[0165] The infected control guinea pigs (Group 11) showed patches
of hair loss and readily visible ulcerated or scaly skin.
Substantially similar lesions were noted in the Group 1 guinea pigs
treated with the placebo, Ex. 5(A), and the Group 2 guinea pigs
treated with the drug-free composition, Ex. 5(B). All of the
terbinafine-containing compositions, Exs. 5(C-J) had clinical
efficacy, based on an improved appearance of the skin as
demonstrated by healthier skin and regrowth of hair in the Groups
3-10 guinea pigs compared to that of guinea pigs treated with the
placebo (vehicle) control and drug-free composition, Exs. 5(A-B).
Clinical efficacy was judged optimized at a drug concentration of
about 1 weight % (Ex. 5(C)) and at a DDAIP penetration enhancer
concentration of about 0.5 weight % (Ex. 5(F)), because increasing
the drug content or increasing the penetration enhancer content did
not provide a further beneficial increase in clinical efficacy.
[0166] At the end of the study, all surviving animals were
sacrificed by an intravenous injection of a euthanasia solution and
disposed to the Animal Resource Center for incineration.
EXAMPLE 6
[0167] This example illustrates in vitro the permeability of a
one-coat type dual action, antifungal nail coat composition
containing terbinafine hydrochloride through hard keratin, using an
animal hoof keratin model (horse hoof) and an agar plate diffusion
assay.
[0168] Three discs, (I, II, and III) were cut from horse hoof
keratin to a thickness in the range of about 0.5 to about 1
millimeter (mm) (Disc I); a thickness in the range of about 1.1 to
about 1.5 mm (Disc II); and thickness in the range of about 1.6 to
about 2 millimeter (mm) (Disc III). The side edges and one face of
each disc was coated with Vaseline to prevent seepage of the
antifungal drug during agar diffusion evaluation leaving the
opposing face exposed.
[0169] In one diffusion assay evaluation, three separate antifungal
coat solutions, 6(A), 6(B), 6(C), were prepared, respectively
containing 25 mg/ml, 0.5 mg/ml, and 1 mg/ml amounts of terbinafine
hydrochloride in dimethylsulfoxide (DMSO). Each antifungal coat
solution was applied to the exposed face of each selected hoof disc
(I, II, and III). The antifungally-coated face of the hoof disc was
then placed in contact with an agar plate seeded with a lawn of
conidial suspension of T. mentagrophytes (ATCC 24953) at a
concentration of 5.times.10.sup.5, and incubated for a period of
about eight hours. The zone of inhibition (diameter of area
remaining clear, i.e., lacking growth) was then measured in
millimeters (mm).
[0170] The results showed that, at all concentrations of
terbinafine hydrochloride, diffusion took place through the hoof
and that the permeate retained bioactivity. The measured zones of
inhibition were generally inversely proportional in diameter to the
thickness of the hoof disc. Hoof disc II having a thickness in the
range of about 1.1 to about 1.5 mm is judged similar to the
thickness of human nails.
EXAMPLE 7
[0171] This example simulates the clinical use of a one-coat type
dual action, antifungal nail coat composition on human nails using
the horse hoof model described in Example 6.
[0172] The general procedure for simulating clinical use is as
follows: The horse hoof is cleaned and washed three times with
buffer. Sections of horse hoof having a thickness of about 100
micrometers are cut using an Arbor blade and are sterilized by
autoclaving. Individual hoof sections are then coated with a
selected nail coat composition containing the amount of terbinafine
hydrochloride and penetration enhancer shown in Compositions 7(A-H)
of Table 5, and left in contact with the nail coat composition. For
comparison, sections of horse hoof are similarly contacted with a
commercial topical nail lacquer, PENLAC.TM. containing the
synthetic antifungal, ciclopirox, (Ex. 7(I)). The treated hoof
sections are then each placed on an agar plate seeded with a lawn
of conidial suspension of T. mentagrophytes (ATCC 24953) at a
concentration of 5.times.10.sup.5, and incubated for a period of
about four days at a temperature of about 35.degree. C. The zone of
inhibition was then measured.
TABLE-US-00005 TABLE 5 Weight Percent Ingredient Terbinafine PVP,
Benzyl Ethanol Example HCl DDAIP.cndot.HCl USP Alcohol to 100% 7(A)
1 None 0.5 0.75 q.s. 7(B) 5 None 0.5 0.75 q.s. 7(C) 10 None 0.5
0.75 q.s. 7(D) 1 0.5 0.5 0.75 q.s. 7(E) 1 2.5 0.5 0.75 q.s. 7(F) 1
5 0.5 0.75 q.s. 7(G) 5 0.5 0.5 0.75 q.s. 7(H) 10 0.5 0.5 0.75 q.s.
7(I) Comparative PENLAC .TM. Nail Lacquer Solution Topical Solution
8% Note: Ex. 7(I) contains 80 mg ciclopirox in a solution base
consisting of ethyl acetate, NF; and butyl monoester of
poly[methylvinylether/maleic acid] in isopropyl alcohol (Dermik
Laboratories, Inc.).
EXAMPLE 8
[0173] This example illustrates the fungicidal activity of one-coat
type dual action, antifungal nail coat compositions containing
terbinafine hydrochloride and, as a penetration enhancer,
DDAIP.HCl, against three strains of the dermatophyte Trichopyton
rubrum (T. rubrum), nine strains of the dermatophyte Trichophyton
mentagrophytes (T. mentagrophytes), and ten strains of the yeast
Candida albicans (C. albicans).
[0174] Nail coat compositions containing terbinafine hydrochloride,
Exs. 8(A), 8(B), and drug free control, Ex. 8(C) were prepared
having the amounts shown in Table 6 and fungicidal efficacy
compared against that of a commercial composition, Ex. 8(D):
PENLAC.TM. Nail Lacquer Solution Topical Solution 8% (Containing
ciclopirox).
TABLE-US-00006 TABLE 6 Weight Percent Ingredient Terbinafine PVP,
Benzyl Ethanol Example HCl DDAIP.cndot.HCl USP Alcohol to 100% 8(A)
1 None 0.5 0.75 q.s. 8(B) 1 0.5 0.5 0.75 q.s. 8(C) None 0.5 0.5
0.75 q.s. 8(D) Comparative PENLAC .TM. Nail Lacquer Solution
Topical Solution 8% Note: Ex. 8(D) contains 80 mg ciclopirox in a
solution base consisting of ethyl acetate, NF; and butyl monoester
of poly[methylvinylether/maleic acid] in isopropyl alcohol (Dermik
Laboratories, Inc.).
[0175] Fungicidal efficacy, based on minimum inhibitory
concentration (MIC) and minimum fungicidal concentration (MFC) of
the drug, was evaluated using a broth microdilution assay, as well
as an agar diffusion plate assay, measuring the zones of
inhibition.
[0176] The broth microdilution method was a modification of a NCCLS
M38-A standard method for the susceptibility testing of
conidium-forming filamentous fungi of the National Committee for
Clinical Laboratory Standards (NCCLS). The modified method was
developed at the Center for Medical Mycology, University Hospitals
of Cleveland, Cleveland, Ohio, based on the method described in
Jessup, et al., "Antifungal Susceptibility Testing of
Dermatophytes: Establishing a Medium for Inducing Conidial Growth
and Evaluation of Susceptibility of Clinical Isolates,"Journal of
Clinical Microbiology, 38, 341-344, published by the American
Society for Microbiology (2000), the disclosures of which are
incorporated herein by reference. Based on a multicenter study of
the reproducibility of the modified method for testing
dermatophytes, adoption of the modified method as an amendment to
the NCCLS M38-A standard has been proposed. The modified method is
described below.
[0177] Dermatophyte isolates are subcultured onto Potato Dextrose
Agar (PDA) and incubated at a temperature of about 30.degree. C.
for a period of about 4 to about 5 days or until good conidiation
is produced. T. rubrum isolates are subcultured onto cereal
(oatmeal) agar instead of PDA in order to induce conidia
production. A suspension of conidia in sterile saline is made by
gently swabbing the colony surface with a sterile swab. The
suspension is allowed to settle for about 5 to about 10 minutes and
the conidia is counted using a hemocytometer. Working suspensions
of conidia are prepared in 10 ml RPMI 1604 (Difco Laboratories)
medium to a final concentration of 1 to 3.times.10.sup.3 CFU/ml.
Yeast controls are subcultured onto PDA and incubated at a
temperature of about 35.degree. C. for about 48 hours. Yeast
inocula are prepared to a final concentration of 0.5 to
2.5.times.10.sup.3 CFU/ml. For MIC assay, each drug concentration
well and growth control well is inoculated with 100 microliters of
cell suspension, and the final volume in each microtiter well is
200 microliters. The dermatophyte plates are incubated at a
temperature of about 35.degree. C. for 4 days (yeast controls for
48 hours). Plates are examined visually for 50% and 80% growth
inhibition as compared to the growth control, and MIC results are
recorded in micrograms (.mu.g)/ml. The MIC endpoint is generally
defined as the lowest concentration that inhibited 80% of fungal
growth as compared to the growth control. To perform the MFC assay,
100 .mu.l is removed from each microtiter well without visible
growth and subcultured onto Potato Dextrose Agar plates. The lowest
concentration to produce <1-2 colonies is considered the MFC.
(Inoculum removed from the microtiter wells is streaked for
isolation--there are no zones of inhibition).
[0178] For the MIC assay, a broth dilution is performed in
microtiter wells with RPMI 1064 as the diluent. The MFC assay is
performed by subculturing the microtiter wells from the MIC
test.
[0179] For the agar diffusion assay, the standardized inoculum of
conidia is applied to the surface of a Potato Dextrose Agar plate
and allowed to dry. Wells are then cut into the agar and the test
composition is put into the wells and allowed to diffuse and
antifungal activity is evidenced by zones of growth inhibition
(i.e., area remaining clear, lacking growth) on the surface of the
plates measured in millimeters (mm) diameter.
[0180] An agar diffusion assay was performed using Potato Dextrose
Agar plates seeded with a lawn of conidial suspension at a
concentration of 5.times.10.sup.5 CFU/ml. The plates were
inoculated separately with undiluted test compositions of Exs.
8(A-D) by adding 200 .mu.l of undiluted test compositions to wells
cut into the agar and allowed to diffuse. The inoculated plates
were then incubated at about 35.degree. C. for 4 days for
dermatophytes and 48 hours for yeast. The range and mean diameter
in millimeters (mm) measurement of the Zone of Inhibition (Zone)
assays of the nail compositions in Table 6 are summarized in Table
6-A below.
TABLE-US-00007 TABLE 6-A Ex. 8(A) Ex. 8(B) Ex. 8(C) Ex. 8(D) Zone
(mm) Zone (mm) Zone (mm) Zone (mm) Organism Range Mean Range Mean
Range Mean Range Mean T. mentagrophytes, n = 9 95-100 97.9 95-100
97.4 13-18 16 30-36 32.2 T. rubrum, n = 3 55-100 84.3 50-98 81
16-18 17.3 30-34 32 C. albicans, n = 10 19-30 23.7 18-30 23.8 0-10
8.5 18-30 25.1
[0181] The data in Table 6-A show that terbinafine-containing nail
coat compositions, Exs. 8(A) and 8(B) were fungicidally active
against all three organisms and substantially equivalent in
activity to one another. The drug-free composition, Ex. 8(C) was
judged substantially ineffective against the yeast, and weakly
effective against the two dermatophytic fungi, indicating that such
activity was likely attributable to antimicrobial effects
contributed by benzyl alcohol and ethanol. The
terbinafine-containing nail coat compositions were judged about
three times more effective against the dermatophytic fungi, T.
mentagrophytes, and T. rubrum, than the commercial
ciclopirox-containing nail lacquer, and were substantially
equivalent to the commercial nail lacquer against the yeast, C.
albicans.
EXAMPLE 9
[0182] The fungicidal activity of terbinafine hydrochloride against
the dermatophytic fungi, T. mentagrophytes, (ATCC 24953), is
illustrated in the modified NCCLS broth dilution assay described in
Example 8, based on minimum inhibitory concentration (MIC) and
minimum fungicidal concentration (MFC) as well as an agar diffusion
plate assay measuring the zones of inhibition.
[0183] A placebo composition, Ex. 9(A), two nail coat compositions
containing terbinafine hydrochloride, Exs. 9(B) and 9(C), and a
drug-free comparative composition, Ex. 9(D) were prepared having
the amounts shown in Table 7. Also prepared were dimethylsulfoxide
(DMSO) solvent solutions of terbinafine hydrochloride, of the
penetration enhancer, DDAIP.HCl, and combinations thereof (Exs.
9(E-H) in the amounts also shown in Table 7. Included for
comparison, was the commercial PENLAC.TM. Nail Lacquer
solution.
TABLE-US-00008 TABLE 7 Weight Percent Ingredient Terbinafine PVP,
Benzyl Ethanol Example HCl DDAIP.cndot.HCl USP Alcohol to 100% 9(A)
None None 0.5 0.75 q.s. (control) 9(B) 1 0.5 0.5 0.75 q.s. 9(C) 1
None 0.5 0.75 q.s. 9(D) None 0.5 0.5 0.75 q.s. 9(E) None 1 mg/ml
None None None in DMSO 9(F) 1 mg/ml in None None None None DMSO
9(G) 1 .mu.g/ml in None None None None DMSO 9(H) 1 .mu.g/ml in 1
.mu.g/ml None None None DMSO in DMSO 9(I) Comparative PENLAC .TM.
Nail Lacquer Solution Topical Solution 8% Note: Ex. 9(I) contains
80 mg ciclopirox in a solution base consisting of ethyl acetate,
NF; isopropyl alcohol, USP; and butyl monoester of
poly[methylvinylether/maleic acid] in isopropyl alcohol (Dermik
Laboratories, Inc.).
[0184] MIC assay was determined using the broth dilution procedure
described in Example 8 performed in microtiter wells with RPMI 1604
as the diluent. Serial dilutions of each test composition were made
in RPMI diluent, and then 100 .mu.l of undiluted test composition
and of each diluted composition was added to a respective
microtiter well. Conidial suspension (100 .mu.l) was then added to
each well and the plates were incubated at an incubation
temperature of about 35.degree. C. for an incubation period of 4
days dermatophytes, and 48 hours for yeasts. For MFC determination,
undiluted test composition was added to wells cut into the agar and
allowed to diffuse. The MIC endpoint was the lowest concentration
that inhibited 80% of fungal growth as compared to the growth
control. The MFC endpoint was the lowest concentration to produce
1-2 colonies. The zone of inhibition size was measured (diameter of
area remaining clear, i.e., lacking growth).
[0185] The zone of inhibition (diameter size in mm), and the
dilution factors for the MIC and MFC assays obtained with each of
the compositions is shown in Table 7-A.
TABLE-US-00009 TABLE 7-A Example Zone size Dilution (Composition:
Diluent) No. (mm) MIC MFC 9(A) Zero 1:32 1:16 9(B) 80 >1:512
>1:512 9(C) 82 >1:512 >1:512 9(D) Zero 1:64 1:32 9(E) Zero
1:32 1:4 9(F) 80 >1:512 >1:512 9(G) 18 0.03 .mu.g/ml 0.125
.mu.g/ml 9(H) 18 0.03 .mu.g/ml 0.125 .mu.g/ml 9(I) 33 >1:512
>1:512
[0186] The terbinafine-containing nail coat compositions, Exs. 9(B)
and 9(C) were fungicidal at the highest dilution (>1:512). The
terbinafine-free compositions, Exs 9(A) and 9(D) were weakly
fungicidal, based on MIC assays, but produced no zone of
inhibition, indicating that any inhibitory effect observed was
likely attributable primarily to some antimicrobial contribution
from the benzyl alcohol and ethanol in the vehicle. The
terbinafine-containing compositions were judged about 2.4 times as
effective as the commercial nail lacquer, Ex. 9(I), at equivalent
volume concentrations, based on the zone of inhibition. The
commercial nail lacquer was comparable to the
terbinafine-containing compositions, based on MIC and MFC assays.
Some difficulty was encountered with the commercial nail lacquer at
the highest concentrations due to evaporation of the lacquer
vehicle and hardening of the lacquer in the microtiter well.
[0187] In DMSO solvent, the fungicidal efficacy at a dilution of
>1:512 of terbinafine hydrochloride at 1 mg/ml concentration was
again confirmed by Ex. 9(F), with at most some weak efficacy from
the penetration enhancer alone (Ex. 9(E)) based on MIC. At a
terbinafine hydrochloride concentration of 1 .mu.g/ml, the
fungicidal efficacy of the terbinafine hydrochloride was
substantially equivalent with or without the penetration enhancer
present (Exs. 9(G), 9(H)).
[0188] The foregoing is intended to be illustrative of the present
invention, but not limiting. Numerous variations and modifications
may be effected without departing from the true spirit and scope of
the invention.
* * * * *