U.S. patent application number 17/688126 was filed with the patent office on 2022-06-23 for method of treating a skin condition.
The applicant listed for this patent is The Procter & Gamble Company. Invention is credited to Teresa DiCOLANDREA, Tomohiro (NMN) HAKOZAKI, Leo Timothy LAUGHLIN, II, Jason Martin WINGET.
Application Number | 20220193061 17/688126 |
Document ID | / |
Family ID | |
Filed Date | 2022-06-23 |
United States Patent
Application |
20220193061 |
Kind Code |
A1 |
HAKOZAKI; Tomohiro (NMN) ;
et al. |
June 23, 2022 |
METHOD OF TREATING A SKIN CONDITION
Abstract
An improved method of treating a skin barrier condition is
disclosed. The method involves identifying a target portion of skin
on a person where treatment is desired and applying a low-pH
composition to the target portion of skin. The low-pH composition
contains an effective amount of a vitamin B.sub.3 compound and has
a pH of less than 5.0.
Inventors: |
HAKOZAKI; Tomohiro (NMN);
(Cincinnati, OH) ; LAUGHLIN, II; Leo Timothy;
(Mason, OH) ; WINGET; Jason Martin; (Mason,
OH) ; DiCOLANDREA; Teresa; (Wyoming, OH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
The Procter & Gamble Company |
Cincinnati |
OH |
US |
|
|
Appl. No.: |
17/688126 |
Filed: |
March 7, 2022 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16460308 |
Jul 2, 2019 |
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17688126 |
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62693613 |
Jul 3, 2018 |
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International
Class: |
A61K 31/455 20060101
A61K031/455; A61P 17/06 20060101 A61P017/06; A61K 45/06 20060101
A61K045/06 |
Claims
1. A method of treating a skin barrier condition, comprising: a)
identifying a target portion of skin on a person where treatment is
desired; and b) applying a low-pH composition to the target portion
of skin during a treatment period, wherein the low-pH composition
comprises an effective amount of niacinamide and has a pH of about
3.5 to less than 4.0 and reduces the amount of S100-A7 [SEQ ID NO:
2] according to the S100-A7 Protein Measurement method.
2. The method of claim 1, wherein the composition comprises about
0.01% to about 10% of the niacinamide.
3. The method of claim 1, wherein the composition downregulates
S100A7 [SEQ ID NO: 1]
4. The method of claim 1, wherein the synergistic reduction in
5100-A7 [SEQ ID NO: 2] is at least 10% more than a predicted
additive amount of S100-A7 [SEQ ID NO: 2] reduction.
5. The method of claim 1, wherein the composition comprises at
least one additional skin care active wherein the at least one
additional skin care active comprises vitamins, minerals, peptides,
sugar amines, sunscreens, oil control agents, flavonoid compounds,
anti-oxidants, protease inhibitors, tyrosinase inhibitors,
anti-inflammatory agents, moisturizing agents, exfoliating agents,
skin lightening agents, anti-acne agents, anti-wrinkle agents,
phytosterols, N-acyl amino acid compounds, antimicrobials,
antifungals, or a combination thereof.
6. The method of claim 1, wherein the composition comprises about
0.1% to about 10% of a stable fatty alcohol thickener.
7. The method of claim 6, wherein the stable fatty alcohol
thickener comprises cetyl alcohols, stearyl alcohols, or a
combination thereof.
8. The method of claim 1, wherein the skin harrier condition is
psoriasis or atopic dermatitis.
9. The method of claim 1, wherein the low-pH composition is free of
neutralized thickeners.
10. A method of treating psoriasis or atopic dermatitis,
comprising: a) determining an effective amount of niacinamide and a
pH for providing a reduction in the expression of 5100 calcium
binding protein A7 [SEQ ID NO: 1] by contacting keratinocytes with
a composition comprising niacinamide and having a pH of about 3.5
to less than 4.0, and measuring the change in the amount of 5100-A7
[SEQ ID NO: 2]; b) combining the effectives amount of niacinamide
with a dermatologically acceptable carrier to make a skin care
composition at the determined pH; c) providing the skin care
composition to a person exhibiting a symptom of psoriasis or atopic
dermatitis; and d) applying the composition to a portion of skin
exhibiting the symptom of psoriasis or atopic dermatitis.
11. The method of claim 10, wherein the amount of niacinamide is
between 0.01% and 10%, by weight of the composition.
12. The method of claim 10, wherein the skin care composition
comprises at least one additional skin care active.
13. The method of claim 10, wherein the composition reduces the
S100-A7 level by at least 10% compared to a corresponding neutral
pH composition.
14. The method of claim 10, wherein the low-pH composition is free
of neutralized thickeners.
Description
FIELD
[0001] The present disclosure is directed generally to a method of
treating the symptoms of a skin barrier condition. More
specifically, the present disclosure is directed to a method of
treating the symptoms of psoriasis and/or atopic dermatitis with an
effective amount of a vitamin B3 compound in a low-pH
composition.
BACKGROUND
[0002] Skin is the first line of defense against environmental
insults that would otherwise damage sensitive underlying tissue and
organs. For example, skin maintains a relatively water-impermeable
barrier between an organism and its environment to prevent
dehydration. Additionally, skin plays a key role in a person's
physical appearance. Generally, most people desire to have healthy
skin that looks healthy and maintains adequate barrier protection.
However, a variety of intrinsic and extrinsic factors can lead to a
decline in skin appearance and barrier function. For example, skin
conditions such as psoriasis and atopic dermatitis (e.g., eczema)
can result in itchy, red, and/or scaly patches of skin that impair
the ability of skin to provide adequate barrier function.
[0003] Numerous agents, both natural and synthetic, are known for
use in skin care compositions marketed to treat various skin
conditions, especially those associated with psoriasis and atopic
dermatitis. One example of a well-known skin care agent used in
cosmetic skin agents is niacinamide. U.S. Pat. No. 5,833,998
discloses the use of niacinamide for regulating the oily/shiny
appearance on skin, and U.S. Pat. No. 5,968,528 discloses the use
of niacinamide for regulating the signs of skin aging.
[0004] In some instances, the combination of niacinamide and other
skin agents has been disclosed. For example, US 2012/0121534 and
U.S. Pat. No. 5,053,230 disclose compositions for promoting the
growth of skin cells to improve the appearance of wrinkled skin.
The compositions in the '534 application and '230 patent are
disclosed as essentially being growth media for stimulating growth
or promoting trophism in skin cells. However, it was not recognized
that using a low pH skin care composition comprising niacinamide
may alleviate symptoms of psoriasis and/or atopic dermatitis,
thereby improving skin barrier function and skin appearance.
[0005] Typically, cosmetic compositions are formulated to have a
slightly acidic to neutral pH (i.e., from 4.0-7.0) which is
believed to improve the stability of certain ingredients in the
composition (e.g., niacinamide, salicylates, and neutralized
thickeners). However, formulating a skin care composition at a
lower pH (e.g., 1.0-4.0) may bolster the acid mantle of the skin,
provide flexibility in other types of skin agents that can be
included in the composition, and/or provide an exfoliation benefit.
Accordingly, it would be desirable to provide a low pH skin care
composition that includes niacinamide for improving skin barrier
function and skin appearance.
SUMMARY
[0006] A method of treating a skin barrier condition is provided
herein. The method involves identifying a target portion of skin on
a person in need of treatment, and then applying a low-pH
composition to the target portion of skin during a treatment
period. The low-pH composition contains an effective amount of a
vitamin B3 compound and has a pH of less than 5.0.
BRIEF DESCRIPTION OF THE DRAWING
[0007] The FIGURE is a bar chart illustrating the synergistic
effect of niacinamide and low pH on the level of normalized S100-A7
[SEQ ID NO: 2].
DETAILED DESCRIPTION
[0008] The undesirable symptoms of psoriasis and atopic dermatitis
are well known. However, it has now been discovered that using
niacinamide at low-pH may be useful for treating symptoms of these
and other skin barrier conditions. Surprisingly, it has also been
discovered that the combination of niacinamide and low pH appears
to provide a synergistic reduction in S100-A7 [SEQ ID NO: 2], which
is a protein believed to play an important role in causing symptoms
associated psoriasis, atopic dermatitis, and other skin barrier
disorders.
[0009] Reference within the specification to "embodiment(s)" or the
like means that a particular material, feature, structure and/or
characteristic described in connection with the embodiment is
included in at least one embodiment, optionally a number of
embodiments, but it does not mean that all embodiments incorporate
the material, feature, structure, and/or characteristic described.
Furthermore, materials, features, structures and/or characteristics
may be combined in any suitable manner across different
embodiments, and materials, features, structures and/or
characteristics may be omitted or substituted from what is
described. Thus, embodiments and aspects described herein may
comprise or be combinable with elements or components of other
embodiments and/or aspects despite not being expressly exemplified
in combination, unless otherwise stated or an incompatibility is
stated.
[0010] In all embodiments, all percentages are by weight of the
cosmetic composition, unless specifically stated otherwise. All
ratios are weight ratios, unless specifically stated otherwise. All
ranges are inclusive and combinable. The number of significant
digits conveys neither a limitation on the indicated amounts nor on
the accuracy of the measurements. All numerical amounts are
understood to be modified by the word "about" unless otherwise
specifically indicated. Unless otherwise indicated, all
measurements are understood to be made at approximately 25.degree.
C. and at ambient conditions, where "ambient conditions" means
conditions under about 1 atmosphere of pressure and at about 50%
relative humidity. All numeric ranges are inclusive of narrower
ranges; delineated upper and lower range limits are interchangeable
to create further ranges not explicitly delineated.
[0011] The compositions of the present invention can comprise,
consist essentially of, or consist of, the essential components as
well as optional ingredients described herein. As used herein,
"consisting essentially of" means that the composition or component
may include additional ingredients, but only if the additional
ingredients do not materially alter the basic and novel
characteristics of the claimed compositions or methods. As used in
the description and the appended claims, the singular forms "a,"
"an," and "the" are intended to include the plural forms as well,
unless the context clearly indicates otherwise.
SEQUENCE LISTING
[0012] A sequence listing that sets forth the nucleotide sequence
for S100 calcium binding protein A7 ("S100A7") [SEQ ID NO: 1] and
protein S100-A7 [SEQ ID NO: 2] are being filed concurrently with
the present application as an ASCII text file titled
"15304P_seq_list_ST25". This ASCII text file was created on Jun.
26, 2018 and is approximately 4.94 KB in size. In accordance with
MPEP .sctn. 605.08 and 37 CFR .sctn. 1.52(e), the subject matter in
the ASCII text file is incorporated herein by reference.
Definitions
[0013] "Apply" or "application", as used in reference to a
composition, means to apply or spread the compositions of the
present invention onto a human skin surface such as the
epidermis.
[0014] "Cosmetic agent" means any substance, as well any component
thereof, intended to be rubbed, poured, sprinkled, sprayed,
introduced into, or otherwise applied to a mammalian body or any
part thereof to provide a cosmetic effect. Cosmetic agents may
include substances that are Generally Recognized as Safe (GRAS) by
the US Food and Drug Administration, food additives, and materials
used in non-cosmetic consumer products including over-the-counter
medications.
[0015] "Effective amount" means an amount of a compound or
composition sufficient to significantly induce a positive benefit
to keratinous tissue over the course of a treatment period. The
positive benefit may be a health, appearance, and/or feel benefit,
including, independently or in combination, the benefits disclosed
herein. In a specific example, an effective amount of a vitamin B3
compound is an amount sufficient to improve the health and/or
appearance of psoriatic skin during a treatment period. In some
instances, an effective amount may be demonstrated using ex vivo
and/or in vitro methods.
[0016] "Improve the appearance of" means providing a measurable,
desirable change or benefit in skin appearance, which may be
quantified, for example, by a decrease in redness, inflammation,
and/or plaque scales.
[0017] "Low pH" means a pH of less than 5.0 (e.g., 1.5 to 5.0
(exclusive); 2.0 to 4.5, 2.5 to 4.0, or about 3.5). A suitable
method of determining the pH of a composition is described in more
detail below.
[0018] "Neutral pH" means a pH of 5.0 to 8.0.
[0019] "Safe and effective amount" means an effective amount of an
ingredient that is low enough to avoid serious side effects (within
the scope of sound medical judgment).
[0020] "Skin care" means regulating and/or improving a skin
condition. Some nonlimiting examples include improving skin
appearance and/or feel by providing a smoother, more even
appearance and/or feel; increasing the thickness of one or more
layers of the skin; improving the elasticity or resiliency of the
skin; improving the firmness of the skin; and reducing the oily,
shiny, and/or dull appearance of skin, improving the hydration
status or moisturization of the skin, improving the appearance of
fine lines and/or wrinkles, improving skin exfoliation or
desquamation, plumping the skin, improving skin barrier properties,
improve skin tone, reducing the appearance of redness or skin
blotches, and/or improving the brightness, radiancy, or
translucency of skin.
[0021] "Skin care active" means a compound or combination of
compounds that, when applied to skin, provide an acute and/or
chronic benefit to skin or a type of cell commonly found therein.
Skin care actives may regulate and/or improve skin or its
associated cells (e.g., improve skin elasticity, hydration, skin
barrier function, and/or cell metabolism).
[0022] "Skin care composition" means a composition that includes a
skin care active and regulates and/or improves skin condition.
[0023] "Synergy" and variations thereof mean a bilirubin degrading
effect provided by using niacinamide in combination with a low-pH
composition that is more than the predicted additive effect of the
vitamin B.sub.3 compound and low pH.
[0024] "Treatment period," as used herein, means the length of time
and/or frequency that a material or composition is applied to a
target skin surface.
[0025] "Vehicle control" means a negative control that is identical
to the test composition except that it does include the particular
active(s) of interest (e.g., does not contain a vitamin B.sub.3
compound).
[0026] Composition
[0027] The skin care compositions herein are intended for topical
application to human skin for improving the appearance and/or
function of psoriatic skin. The compositions herein include an
effective amount of a vitamin B.sub.3 compound and have a pH of
less than 5.0 (e.g., less than 4.5, 4.0, 3.5, 3.0, 2.5 or even 2.0
or less). The compositions are formed by mixing the vitamin B.sub.3
compound with a dermatologically acceptable carrier, which may be
done using conventional methods known to those skilled in the art.
The compositions may optionally include one or more skin actives of
the type commonly included in skin care compositions of the type.
The compositions may be cosmetic compositions, pharmaceutical
compositions, or cosmeceutical compositions, and may be provided in
various product forms, including, but are not limited to,
solutions, suspensions, lotions, creams, gels, toners, sticks,
sprays, aerosols, ointments, cleansing liquid washes and solid
bars, pastes, foams, mousses, shaving creams, wipes, strips,
patches, electrically-powered patches, hydrogels, film-forming
products, facial and skin masks (with and without insoluble sheet),
and the like. The composition form may follow from the particular
dermatologically acceptable carrier chosen, if present in the
composition.
[0028] Vitamin B.sub.3 Compound
[0029] The compositions of the present invention include a safe and
effective amount of a vitamin B.sub.3 compound. In addition to
treating one or more symptoms of psoriasis and/or atopic
dermatitis, the vitamin B.sub.3 compound may also be useful for
regulating other skin condition, for example, as described in U.S.
Pat. No. 5,939,082. The compositions herein may contain 0.01% to
15%, by weight, of the vitamin B.sub.3 compound, based on the
weight or volume of the composition (e.g., 0.1% to 10%, 0.1% to 3%,
0.5% to 8%, 1% to 5%, or even 2% to 4%).
[0030] As used herein, "vitamin B.sub.3 compound" means a compound
having the formula:
##STR00001##
Where:
[0031] R is CONH.sub.2 (i.e., niacinamide), COOH (i.e., nicotinic
acid) or CH.sub.2OH (i.e., nicotinyl alcohol); derivatives thereof;
and salts of any of the foregoing.
[0032] Exemplary derivatives of vitamin B.sub.3 compounds include
nicotinic acid esters, including non-vasodilating esters of
nicotinic acid (e.g., tocopheryl nicotinate, myristyl nicotinate)
nicotinamide riboside, nicotinyl amino acids, nicotinyl alcohol
esters of carboxylic acids, nicotinic acid N-oxide, and niacinamide
N-oxide.
[0033] Dermatologically Acceptable Carrier
[0034] The compositions herein include a dermatologically
acceptable carrier (which may be referred to as a "carrier"). The
phrase "dermatologically acceptable carrier" means that the carrier
is suitable for topical application to the keratinous tissue, has
good aesthetic properties, is compatible with the actives in the
composition, and will not cause any unreasonable safety or toxicity
concerns. In one embodiment, the carrier is present at a level of
from about 50% to about 99%, about 60% to about 98%, about 70% to
about 98%, or, alternatively, from about 80% to about 95%, by
weight of the composition.
[0035] The carrier can be in a wide variety of forms. In some
instances, the solubility or dispersibility of the components
(e.g., extracts, sunscreen active, additional components) may
dictate the form and character of the carrier. Non-limiting
examples include simple solutions (e.g., aqueous or anhydrous),
dispersions, emulsions, and solid forms (e.g., gels, sticks,
flowable solids, or amorphous materials). In some instances, the
dermatologically acceptable carrier is in the form of an emulsion.
The emulsion may have a continuous aqueous phase (e.g., an
oil-in-water or water-in-oil-in-water emulsion) or a continuous oil
phase (e.g., water-in-oil or oil-in-water-in-oil emulsion). The oil
phase of the present invention may comprise silicone oils,
non-silicone oils such as hydrocarbon oils, esters, ethers, and
mixtures thereof. The aqueous phase typically comprises water and
water-soluble ingredients (e.g., water-soluble moisturizing agents,
conditioning agents, anti-microbials, humectants and/or other skin
care actives). However, in some instances, the aqueous phase may
comprise components other than water, including but not limited to
water-soluble moisturizing agents, conditioning agents,
anti-microbials, humectants and/or other water-soluble skin care
actives. In some instances, the non-water component of the
composition comprises a humectant such as glycerin and/or other
polyol(s).
[0036] In some instances, the compositions herein are in the form
of an oil-in-water ("O/W") emulsion that provides a sensorial feel
that is light and non-greasy. Suitable O/W emulsions herein may
include a continuous aqueous phase of more than 50% by weight of
the composition, and the remainder being the dispersed oil phase.
The aqueous phase may include 1% to 99% water, based on the weight
of the aqueous phase, along with any water soluble and/or water
miscible ingredients. In these instances, the dispersed oil phase
will typically be present at less than 30% by weight of composition
(e.g., 1% to 20%, 2% to 15%, 3% to 12%, 4% to 10%, or even 5% to
8%) to help avoid some of the undesirable feel effects of oily
compositions. The oil phase may include one or more volatile and/or
non-volatile oils (e.g., botanical oils, silicone oils, and/or
hydrocarbon oils). Some nonlimiting examples of oils that may be
suitable for use in the present compositions are disclosed in U.S.
Pat. No. 9,446,265 and U.S. Publication No. 2015/0196464.
[0037] The carrier may contain one or more dermatologically
acceptable, hydrophilic diluents. As used herein, "diluent"
includes materials in which the vitamin B.sub.3 compound can be
dispersed, dissolved, or otherwise incorporated. Hydrophilic
diluents include water, organic hydrophilic diluents such as lower
monovalent alcohols (e.g., C.sub.1-C.sub.4) and low molecular
weight glycols and polyols, including propylene glycol,
polyethylene glycol (e.g., molecular weight of 200 to 600 g/mole),
polypropylene glycol (e.g., molecular weight of 425 to 202.5
g/mole), glycerol, butylene glycol, 1,2,4-butanetriol, sorbitol
esters, 1,2,6-hexanetriol, ethanol, isopropanol, sorbitol esters,
butanediol, ether propanol, ethoxylated ethers, propoxylated ethers
and combinations thereof.
[0038] Emulsifier
[0039] When the dermatologically acceptable carrier is in the form
of an emulsion, it may be desirable to include art emulsifier to
provide a stable composition (e.g., does not phase separate). When
included, the emulsifier may be present at an amount of 0.1% to 10%
(e.g., 1% to 5%, or 2%-4%). Emulsifiers may be nonionic, anionic or
cationic. Some non-limiting examples of emulsifiers that may be
suitable for use herein are disclosed in U.S. Pat. Nos. 3,755,560;
4,421,769; and McCutcheon's Detergents and Emulsifiers, North
American Edition, pages 317-324 (1986).
[0040] Thickeners
[0041] In some instances, it may be desirable to use thickeners
that tolerate a lower range of pH. For example, neutralized
thickeners may degrade at lower pH and thus may not impart the
desired thickening or feel properties to the composition. On the
other hand, fatty alcohol thickeners such as cetyl alcohols and
stearyl alcohols are generally stable at low pH (e.g., pH of less
than 5.0 or even between a pH of about 2.5 to about 4.0), and thus
may be particularly suited for use in the low pH compositions
herein. Accordingly, the present compositions may be free or
substantially free of neutralized thickeners and/or may have from
0.1% to 10% (e.g., from about 0.5% to about 8%, from about 1.0% to
about 5%, or even from about 2% to about 4%) of a fatty alcohol
thickener.
[0042] Other Optional Ingredients.
[0043] The present composition may optionally include one or more
additional ingredients commonly used in cosmetic compositions
(e.g., colorants, skin care actives, anti-inflammatory agents,
sunscreen agents, emulsifiers, buffers, rheology modifiers,
combinations of these and the like), provided that the additional
ingredients do not undesirably alter the skin health or appearance
benefits provided by the present compositions. The additional
ingredients, when incorporated into the composition, should be
suitable for use in contact with human skin tissue without undue
toxicity, incompatibility, instability, allergic response, and the
like. Some nonlimiting examples of additional actives include
vitamins, minerals, peptides and peptide derivatives, sugar amines,
sunscreens, oil control agents, particulates, flavonoid compounds,
hair growth regulators, anti-oxidants and/or anti-oxidant
precursors, preservatives, protease inhibitors, tyrosinase
inhibitors, anti-inflammatory agents, moisturizing agents,
exfoliating agents, skin lightening agents, sunless tanning agents,
lubricants, anti-acne actives, anti-cellulite actives, chelating
agents, anti-wrinkle actives, anti-atrophy actives, phytosterols
and/or plant hormones, N-acyl amino acid compounds, antimicrobials,
and antifungals. Other non-limiting examples of additional
ingredients and/or skin care actives that may be suitable for use
herein are described in U.S. Publication Nos. 2002/0022040;
2003/0049212; 2004/0175347; 2006/0275237; 2007/0196344;
2008/0181956; 2008/0206373; 2010/00092408; 2008/0206373;
2010/0239510; 2010/0189669; 2010/0272667; 2011/0262025;
2011/0097286; US2012/0197016; 2012/0128683; 2012/0148515;
2012/0156146; and 2013/0022557; and U.S. Pat. Nos. 5,939,082;
5,872,112; 6,492,326; 6,696,049; 6,524,598; 5,972,359; and
6,174,533.
[0044] When including optional ingredients in the compositions
herein, it may be desirable to select ingredients that do not form
complexes or otherwise undesirably interact with other ingredients
in the composition at low pH, especially pH sensitive ingredients
like niacinamide, salicylates and peptides. In some instances, it
may be desirable to select skin care actives that function via
different biological pathways so that the actives do not interfere
with one another, which could reduce the efficacy of both agents.
When present, the optional ingredients may be included at amounts
of from 0.0001% to 50%; from 0.001% to 20%; or even from 0.01% to
10% (e.g., 50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5% or
0.1%), by weight of the composition.
Methods of Use
[0045] The low-pH compositions herein include an effective amount
of a vitamin B.sub.3 compound and are formulated for topical
application to skin. The method involves identifying a target
portion of skin on a person in need of treatment or where treatment
is desired (e.g., skin that is exhibiting impaired barrier
function) and applying the low-pH composition to the target portion
of skin. The target portion of skin may be on a facial skin surface
such as the forehead, perioral, chin, periorbital, nose, and/or
cheek) or another part of the body (e.g., hands, arms, legs, back,
chest). The target portion of skin may be identified according to
known methods of identifying skin with impaired barrier function.
For example, the target portion of skin may be identified as
needing treatment if it exhibits signs of psoriasis or atopic
dermatitis (redness, itchiness, painfulness, inflammation, plaques,
scales, etc.). In another example, the target portion of skin may
be identified as needing treatment if it exhibits a trans-epidermal
water loss (TEWL) that exceeds a threshold level. In still another
example, the target portion of skin may be identified as needing
treatment if expression of S100A7 [SEQ ID NO: 1] is greater than a
threshold value, for example, as demonstrated by a high level of
S100-A7 [SEQ ID NO: 2]. In a further example, a target portion of
skin may be selected that does not currently exhibit signs of
reduced barrier function, but the user desires to provide a
preventative benefit, especially if the target portion of skin has
previously exhibited signs of reduced barrier function.
[0046] In some instances, the method of treating a skin barrier
condition involves selecting an effective amount of a vitamin
B.sub.3 compound and a suitable pH for a skin care composition that
will provide a synergistic reduction in S100-A7 [SEQ ID NO: 2]. The
selected amount of vitamin B.sub.3 can be combined with a
dermatologically acceptable carrier to make the skin care
composition at the selected pH. The resulting skin composition is
then provided to person in need of treatment, for example, via a
retail store or direct-to-consumer sale. In this example, the
effective amount of vitamin B3 compound and/or pH of the skin
composition may be selected by determining their ability to
synergistically reduce S100-A7 [SEQ ID NO: 2] in at least one of an
in vitro assay, ex vivo assay, of in vivo assay.
[0047] The composition may be applied locally to the target portion
of skin in need of treatment and, if desired, to the surrounding
skin at least once a day, twice a day, or on a more frequent daily
basis, during a treatment period. When applied twice daily, the
first and second applications are separated by at least 1 to 12
hours. Typically, the composition is applied in the morning and/or
in the evening before bed. When used according to the methods
herein, the present compositions may improve the appearance and/or
barrier function of skin, for example, by reducing redness,
inflammation, itchiness, pain, dry/flaky skin, trans-epidermal
water loss, the size and/or number of psoriatic plaques, and/or the
level of S100-A7 [SEQ ID NO: 2]).
[0048] The treatment period is ideally of sufficient time for the
vitamin B.sub.3 compound present in the low-pH composition to
improve the appearance and/or barrier function of a target portion
of skin. The treatment period typically lasts for at least 1 week
(e.g., about 2 weeks, 4 weeks, 8 weeks, or even 12 weeks). In some
instances, the treatment period may extend over multiple months
(i.e., 3-12 months). In some instances, the composition is applied
most days of the week (e.g., at least 4, 5 or 6 days a week), at
least once a day or even twice a day during a treatment period of
at least 2 weeks, 4 weeks, 8 weeks, or 12 weeks.
[0049] The step of applying the composition may be accomplished by
localized application. In reference to application of the
composition, the terms "localized", "local", or "locally" mean that
the composition is delivered to the targeted area (e.g., a
psoriatic plaque) while minimizing delivery to skin surfaces where
treatment is not desired. The composition may be applied and
lightly massaged into an area of skin. The form of the composition
or the dermatologically acceptable carrier should be selected to
facilitate localized application. While certain embodiments herein
contemplate applying a composition locally to an area, it will be
appreciated that compositions herein can be applied more generally
or broadly to one or more skin surfaces. In certain embodiments,
the compositions herein may be used as part of a multi-step beauty
regimen, wherein the present composition may be applied before
and/or after one or more other compositions.
EXAMPLE
[0050] This example demonstrates the unexpected ability of a low-pH
composition comprising niacinamide to potentially improve the
appearance and/or barrier function of skin suffering from
psoriasis, atopic dermatitis, and/or skin barrier conditions. In
this example, changes in the level of S100-A7 [SEQ ID NO: 2] are
used as a surrogate for the effect of the low-pH, niacinamide
containing composition on skin barrier function.
[0051] Skin care agents that can downregulate S100A7 [SEQ ID NO: 1]
are considered a promising approach to treating symptoms of
psoriasis, atopic dermatitis, and other skin barrier conditions.
S100A7 [SEQ ID NO: 1], also called psoriasin, is a member of the
S100 multigene family that is encoded in the epidermal
differentiation complex on chromosome 1q21. S100A7 [SEQ ID NO: 1]
is highly expressed in epidermal hyperproliferative disease, and
overexpression of S100A7 [SEQ ID NO: 1] is believed to contribute
to the symptoms of psoriasis (Ekman, et al., "Overexpression of
Psoriasin (S100A7) Contributes to Dysregulated Differentiation in
Psoriasis," Acta Derm Venereol, 2017 Apr. 6, 97(4); 441-448).
S100A7 [SEQ ID NO: 1] has also been reported to be upregulated in
cases of atopic dermatitis and other skin barrier conditions
(Glaser, et al., (2008); "The Antimicrobial Protein Psoriasin
(S100A7) Is Upregulated in Atopic Dermatitis and after Experimental
Skin Barrier Disruption" Journal of Investigative Dermatology,
129(3), 641-649). Thus, it is believed, without being limited by
theory, that inhibiting the expression of S100A7 [SEQ ID NO: 1]
and/or reducing the level of S100-A7 [SEQ ID NO: 2] will alleviate
symptoms of psoriasis, atopic dermatitis, and other skin barrier
conditions associated with overexpression of S100A7 [SEQ ID NO: 1]
and/or accumulation of 5100-A7 [SEQ ID NO: 2].
[0052] In this example, a low-pH composition (pH 2.5) comprising
niacinamide at 5% (w/v) was tested to determine its ability to
regulate the expression of S100A7 [SEQ ID NO: 1]. A vehicle control
was used in this Example as the negative control. The expression
level of S100A7 [SEQ ID NO: 1] was determined by measuring the
normalized amount of protein S100-A7 [SEQ ID NO: 2] present in each
sample.
Sample Prep and Test Method
[0053] Keratinocytes from human donors (available from Lonza, N.J.)
are cultivated with Complete KBM Gold media until they reached
70-80% confluency. The keratinocytes are then subcultured per
manufacturer's recommendations and used at either passage 1 or 2.
For growth of keratinocytes on de-epidermized dermis (DED), two
media are used. Medium 1 is used for the first three days while the
cultures remained submerged and Medium 2 is used when cultures are
raised to the air-liquid interface and then until the time of
collection.
[0054] Medium 1 consists of: Dulbecco's Modified Eagle Medium
(DMEM) and Ham's F-12 Nutrient Mixture at a ratio of 3:1, followed
by the addition of Hyclone Cosmic Calf Serum (5%), Hydrocortisone
(0.4 .mu.g/ml), epidermal growth factor (0.02 mg/ml), transferrin
(3 mg/me, insulin (5 .mu.g/ml), cholera toxin (0.02 .mu.g/ml),
triiodothyronine (2.times.10.sup.-11 M), adenine (0.18 mM), sodium
pyruvate 1.times., GlutaMax 1.times. (Invitrogen), CaCl.sub.2 (300
uM), 1.times.CD lipid concentrate 300 .mu.M, fibroblast growth
factor 7 (FGF-7) (10 ng/ml), and penicillin/streptomycin
1.times..
[0055] Medium 2 consists of: medium 1 modified with the addition of
1% serum and removal of FGF-7 and 1 mM CaCl.sub.2. Medium 1 is used
for two days while the cultures remain submerged and Medium 2 is
used for cultures raised to the air-liquid interface.
[0056] De-epidermized dermis (DED) is prepared by removing fat from
the skin sample with a scalpel, cutting the skin into squares
measuring 1.25 cm.sup.2, and placing the samples in 1M NaCl plus
10.times. penicillin/streptomycin. The sample is incubated
overnight at 37.degree. C. The following day, the epidermis is
carefully peeled off with forceps and dermal tissue is stored in
phosphate-buffered saline (PBS) plus 2.times.
penicillin/streptomycin at 4.degree. C. until ready for use.
[0057] Approximately 5.times.10.sup.5 keratinocytes in 50 .mu.l of
Medium 1 are pipetted into 8 mm cloning cylinders placed atop DEDs.
2 ml of Medium 1 is added to the bottom of the 6-well plate
containing the transwell. The plates are incubated overnight at
33.degree. C. in 5% CO.sub.2 and 55% RH. The following day, the
cloning cylinders are removed and cultures are submerged in Medium
1. At three days, cultures are raised to the air-liquid interface
in Medium 2.
[0058] At day 7 at the air-liquid interface, the cultures are
treated topically with one of four test compositions (a vehicle
control at pH 2.5 or pH 5, or a 5% (w/v) niacinamide composition at
pH 2.5 or pH 5), and one sample is left untreated, for a total of
five test legs. Each test leg has four replicates. The pH of each
sample can be adjusted using 1M HCl pH titration, for example,
performed separately before the experiment set up to determine how
much 1M HCl is needed for each leg to achieve targeting pH level.
Cultures are dosed topically twice per day for 5 days. The
keratinocytes are isolated from each culture on day 5 using 3.8%
ammonium thiocyanate. The keratinocytes are weighed and then
flash-frozen for mass spectrometry and protein level
determination.
S100-A7 Protein Measurement
[0059] Frozen keratinocyte samples are thawed. 20 ul/mg (wet
weight) of keratinocytes are placed in a solution of 4% sodium
dodecyl sulphate, 0.1 M dithiothreitol, and 150 mM Tris (pH 7.5)
and sonicated for 30 minutes to lyse the cells. Filter-aided sample
preparation is used to prepare peptides for Liquid
Chromatography-Mass Spec analysis. Lysate prepared above is
centrifuged for 10 minutes at 18,000.times.g to clarify, and the
supernatant is then mixed in a 1:10 ratio with 180 .mu.l 8M Urea,
150 mM Tris (pH 8.5). This mixture is placed into YM-30 centrifugal
filter unit (Millipore) and centrifuged for 10 minutes at
14,000.times.g. Proteins are captured on the filter. We then add
200 .mu.l 8M Urea, 150 mM Tris pH 8.5 to the filter and spin as
above to wash. 100 .mu.l 50 mM iodoacetamide is added to the filter
and incubated for 30 minutes at ambient temperature in the dark to
alkylate cysteine residues. This buffer is then centrifuged through
the filter and followed with 3 wash steps of 200 .mu.l 8M Urea, 150
mM Tris pH 8.5 as above. This is followed by 3 wash steps of 200
.mu.l 100 mM Ammonium bicarbonate. 40 .mu.l of 0.5 .mu.g/.mu.l
Trypsin/LysC (Promega) in 100 mM Ammonium bicarbonate is added to
the filter. Filter units are then incubated 16 hours in a
37.degree. C. incubator. Resulting peptides are collected in clean
tubes by centrifugation as above. Synthetic isotopically labeled
internal standards (e.g., from New England Peptide) are added in
equal amounts to each sample. These standards include the peptide
sequence GTNYLADVFEK, with the C-terminal lysine labeled using
13C(6)15N(2). Peptides are analyzed using an Agilent 1690 Infinity
LC system coupled to an Agilent 6490 QQQ mass spectrometer (or
equivalent) against a scheduled multiple reaction monitoring
method. Peptides are loaded onto an Agilent Zorbax RRHD Eclipse
Plus 95 .ANG. C18, 2.1.times.150 mm, 1.8 .mu.m, 1200 bar column
heated to 50.degree. C. Mobile phases are A: 0.1% Formic acid in
water and B: 0.1% Formic Acid, 90% Acetonitrile, 9.9% water. The
flow rate is 0.4 ml/min. The dynamic MRM method contains 4
transitions for the GTNYLADVFEK peptide. These transitions are
based on the doubly-charged precursor 628.8115 m/z for the
endogenous peptide and 632.8186 m/z for the labeled internal
standard. Peaks are manually verified using the Skyline software
and peak areas are exported for further normalization and
analysis.
[0060] The results of the test are summarized below in Table 1. The
protein values shown are the averaged amount of S100-A7 [SEQ ID NO:
2] detected for the 4 replicates in each test leg. The measured
protein amount is the log 2 normalized intensity after fitting
nonlinear regression models using the default settings for the
MSstats statistical analysis package. The delta versus untreated
value is calculated by subtracting the measured protein amount for
each leg from the measured protein amount for the untreated leg.
The normalized protein amount reflects the amount of measured
S100-A7 normalized to the untreated sample, which is assigned a
baseline value of 100. The p-value is determined by combined
pairwise ANOVA comparison of the experimental conditions corrected
for multiple hypothesis testing using the Benjamini & Hochberg
method. A p-value of 0.1 or less is considered statistically
significant.
TABLE-US-00001 TABLE 1 Measured protein .DELTA. vs. Normalized
Sample (log2) Untreated protein p-value Untreated 18.75 0 100
Vehicle pH 2.5 17.8 0.95 51.8 0.2057 Vehicle pH 5 17.6 1.15 45
0.0496 5% Niacinamide at pH 2.5 15.5 3.25 10.5 <0.0001 5%
Niacinamide at pH 5 17.5 1.25 42 0.0613
[0061] Table 2 and FIG. 1, illustrate the unexpected reduction in
S100-A7 [SEQ ID NO: 2] caused by the low-pH, 5% niacinamide
composition. The protein amounts provided in Table 2 are based on
the normalized protein amounts from Table 1. As can be seen in
Table 2 and FIG. 1, lowering the pH of the vehicle control does not
result in a statistically significant change in protein level. The
5% niacinamide composition also appears to have no significant
effect on protein level at neutral pH. Based on the observed
individual effects of low pH and niacinamide at neutral pH, it
would be expected that the combined effect would not significantly
affect the level of S100-A7 [SEQ ID NO: 2]. However, the observed
effect of using niacinamide in a low-pH composition surprisingly
resulted in a significant change in the level of S100-A7 [SEQ ID
NO: 2]. Thus, providing a low-pH skin care composition that
includes a vitamin B.sub.3 compound such as niacinamide may provide
a better way to treat skin barrier conditions related to an
overexpression of S100A7 [SEQ ID NO: 1].
TABLE-US-00002 TABLE 2 .DELTA. Normalized Test Leg Comparison
Protein p-value A. Effect of Niacinamide 3 0.9823 (Vehicle pH 5-5%
Niacinamide pH 5) B. Effect of low pH -6.8 0.8789 (Vehicle pH
5-Vehicle pH 2.5) C. Observed effect of low pH + Niacinamide 34.5
<0.0005 D. Expected effect of low pH + 5% -3.8 0.7011
Niacinamide (A + B)
[0062] The dimensions and values disclosed herein are not to be
understood as being strictly limited to the exact numerical values
recited. Instead, unless otherwise specified, each such dimension
is intended to mean both the recited value and a functionally
equivalent range surrounding that value. For example, a dimension
disclosed as "40 mm" is intended to mean "about 40 mm".
[0063] Every document cited herein, including any cross referenced
or related patent or application and any patent application or
patent to which this application claims priority or benefit
thereof, is hereby incorporated herein by reference in its entirety
unless expressly excluded or otherwise limited. The citation of any
document is not an admission that it is prior art with respect to
any invention disclosed or claimed herein or that it alone, or in
any combination with any other reference or references, teaches,
suggests or discloses any such invention. Further, to the extent
that any meaning or definition of a term in this document conflicts
with any meaning or definition of the same term in a document
incorporated by reference, the meaning or definition assigned to
that term in this document shall govern.
[0064] While particular embodiments of the present invention have
been illustrated and described, it would be obvious to those
skilled in the art that various other changes and modifications can
be made without departing from the spirit and scope of the
invention. It is therefore intended to cover in the appended claims
all such changes and modifications that are within the scope of
this invention.
Sequence CWU 1
1
312918DNAHomo sapiens 1gtccaaacac acacatctca ctcatccttc tactcgtgac
gcttcccagc tctggtaagt 60ctcacctgcc tctttgcgtt ttctagaagt gcccagtgcc
tggcctgcca tgctgcctgt 120aggaagggaa agggctgtgg acaaagactc
agaagcctgg gtctaggcca gctctgccat 180tagctgggct gtgtgacctt
cacttgaccc caaaagccca cgtctgtata accaggtggc 240tgaaccagac
ctgtccattg ctcctttggg atctcctgag gcaactcctg tggaaatgtc
300cccagtggga cctgagcaca ggaccctccc tgtcctcttc ctccctggat
tcatcctcct 360gttggggtcc cggttgctaa atcccctccc cagctccagc
tccacagagg gtggagggct 420aggggtcccc tgtgaatgct ttggaggctc
tccaaacccc caggcctggg accccgacac 480agcatagccc tggccagatg
gcttgcccct acctgggtaa ggtattgaga aaccaacgct 540ccgtcactcc
ctaccacaac ctgcaaattg tcagtcaccc tctgggcaca gggcttgccc
600tctggccttg gcctcagatt caaaaaagct ctgggtggct cctcttagtc
agttgccggc 660atggagcttt ctcctgcatt gagaggtgtt gggggtttat
tgtcagacac ccatatgcat 720tagccaaggt cctttcctgg ttctttggaa
agtgtgggaa gtgagaactg cactgtccta 780tttttcaggc ttggggtgca
ggccagccca gactgactac tggctgaaag atctgctttg 840ggaagtcaga
catttttatt taaacagtca aggcttggtg attcagaggg tgcagataat
900tcccaagagc agacatgaag tgactcccac ctcactccca ccccgtccag
cagcttaaga 960accagagagt tatttctgtt ccttgtaaca gaggccattc
ctgtttctcg ttcccaaggg 1020aatacaagaa gggccccctg tgctcgggac
aggactgtgt ttcctgctaa cactcacgtc 1080cacgctcaca tgtgcacctg
gagcctgcac acaccgtgtc atgcagggct tgcacttgag 1140tcaaggtggg
gcccacggtg tctggttcct gaagcagccg aggagggtca gacaccagat
1200caccaaggcc attgtcctgc ccccatgact gtcaccccga ctgtgaagct
ggaggagatg 1260gatttggtgc tcttttaaga gtggggtctg ccatgtgctg
gctgaggctg gggtgtccca 1320gcttagttct tccatgtgtt gctggcgccc
gactctccct gcttctccct ctctgaagca 1380ctgtccacag ctggacttcc
gccatccacc tgcatctcct cttggccctg gccaggatgg 1440gccaggtctc
agacttggtc ctaccctctc atttttgagc aacttatccc atcacattaa
1500aaaaaaattg agttcgttct gctacacctt atccgtgttt ttactctgtc
ctcagccctc 1560cttccagcac tcccaaatag agaccctaac ttaaatgaag
ggaaaaaaat gattgtcttt 1620atttcctgaa ggctttttga aagcaaagat
gagcaacact caagctgaga ggtccataat 1680aggcatgatc gacatgtttc
acaaatacac cagacgtgat gacaagattg agaagccaag 1740cctgctgacg
atgatgaagg agaacttccc caacttcctt agtgcctgtg tgagtcggtg
1800tctagcttct caatgttgga ggatacattt tgctatgtgg ccttggacag
atcaccctca 1860ccctctgatt aaaaaatttc ccatttgcaa atagggcttt
attacttgga tcatatgtga 1920atctaaggat ggtaggcaaa aaagtatgaa
aatgggaaag agttatacag atataaagga 1980ggttattaga tggtcagcaa
agtgtgcagc ttgagggcag tgcacagtcc cctcccagca 2040ctcactgacc
ctctctctgt gagactgaaa ctcacatgct caggtcctgt ttcccaaagg
2100ccccacatca acatccctga gattactggg aaagcactag aagttccatt
gttagtagaa 2160atcaatagcc ctctttgcaa taaacaagac tcattagcaa
atctgaaaaa aaactgtggg 2220ctactaggta ccaaagggtc tagatgacca
agagtaggat acccagaaga tgagtttggt 2280ggaaaaggag aagaaaggag
aggaggccag agaaagaaga gaacaaaagc atgagggcca 2340attttgaggc
cctggggtag aactaaggta ggcagtgccc ttagaggctg ggacgggaaa
2400gagtgaggct ggggcagggg actgctctcc ccaaggtcac ttaacagagc
accttgggac 2460aggacctgcc tcccatcctg aggtgtgaga caaaaagact
ccctagagaa ctccagggat 2520aacactcacg tcctcacccc aacttcaccc
actcaccctg tgctctcagc cccgccacca 2580tgcctgtgca gatctaggta
cttgtctgta tctctgcctc ctcctctccc tcccagccca 2640aaacttgttt
gtgattgaat ttttctattt tgtatgtttt tctcttcaca ggacaaaaag
2700ggcacaaatt acctcgccga tgtctttgag aaaaaggaca agaatgagga
taagaagatt 2760gatttttctg agtttctgtc cttgctggga gacatagcca
cagactacca caagcagagc 2820catggagcag cgccctgttc cgggggcagc
cagtgaccca gccccaccaa tgggcctcca 2880gagaccccag gaacaataaa
atgtcttctc ccaccaga 29182101PRTHomo sapiens 2Met Ser Asn Thr Gln
Ala Glu Arg Ser Ile Ile Gly Met Ile Asp Met1 5 10 15Phe His Lys Tyr
Thr Arg Arg Asp Asp Lys Ile Glu Lys Pro Ser Leu 20 25 30Leu Thr Met
Met Lys Glu Asn Phe Pro Asn Phe Leu Ser Ala Cys Asp 35 40 45Lys Lys
Gly Thr Asn Tyr Leu Ala Asp Val Phe Glu Lys Lys Asp Lys 50 55 60Asn
Glu Asp Lys Lys Ile Asp Phe Ser Glu Phe Leu Ser Leu Leu Gly65 70 75
80Asp Ile Ala Thr Asp Tyr His Lys Gln Ser His Gly Ala Ala Pro Cys
85 90 95Ser Gly Gly Ser Gln 100311PRTArtificial
SequenceCommercially available amino acid sequence used as an
internal standard for protein measurement 3Gly Thr Asn Tyr Leu Ala
Asp Val Phe Glu Lys1 5 10
* * * * *