U.S. patent application number 15/304818 was filed with the patent office on 2018-12-06 for topical compositions and methods for reducing oxidative stress.
The applicant listed for this patent is Kimberly-Clark Worldwide, Inc.. Invention is credited to Josh Gregorio, Seungjun 'Jason' Lee, Debbie Ngai, Lucia Piccotti, Penkanok Sriwiriyanont, David J. Tyrrell, Rebecca Uelmen, Scott W. Wenzel.
Application Number | 20180344622 15/304818 |
Document ID | / |
Family ID | 54359470 |
Filed Date | 2018-12-06 |
United States Patent
Application |
20180344622 |
Kind Code |
A9 |
Piccotti; Lucia ; et
al. |
December 6, 2018 |
TOPICAL COMPOSITIONS AND METHODS FOR REDUCING OXIDATIVE STRESS
Abstract
Compositions for reducing skin aging resulting from oxidative
stress and/or photodamage are disclosed herein. The compositions
can be topically applied to a skin region to reduce or prevent skin
wrinkles, fine lines, thinning skin, sagging skin, skin dryness,
and skin itchiness.
Inventors: |
Piccotti; Lucia; (Houston,
TX) ; Tyrrell; David J.; (Milwaukee, WI) ;
Sriwiriyanont; Penkanok; (Neenah, WI) ; Ngai;
Debbie; (Neenah, WI) ; Gregorio; Josh;
(Neenah, WI) ; Wenzel; Scott W.; (Neenah, WI)
; Uelmen; Rebecca; (Greenville, WI) ; Lee;
Seungjun 'Jason'; (Neenah, WI) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Kimberly-Clark Worldwide, Inc. |
Neenah |
WI |
US |
|
|
Prior
Publication: |
|
Document Identifier |
Publication Date |
|
US 20170042799 A1 |
|
|
US 20180064633 A9 |
March 8, 2018 |
|
|
Family ID: |
54359470 |
Appl. No.: |
15/304818 |
Filed: |
April 30, 2014 |
PCT Filed: |
April 30, 2014 |
PCT NO: |
PCT/US2014/036199 PCKC 00 |
371 Date: |
October 17, 2016 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61Q 19/08 20130101;
A61K 36/28 20130101; A61Q 19/004 20130101; A61K 36/03 20130101;
A61K 9/0014 20130101; A61K 2800/10 20130101; A61K 8/9711 20170801;
A61P 17/00 20180101; A61K 8/9789 20170801; A61K 2800/522 20130101;
A61K 36/28 20130101; A61K 2300/00 20130101; A61K 36/03 20130101;
A61K 2300/00 20130101 |
International
Class: |
A61K 8/97 20060101
A61K008/97; A61Q 19/08 20060101 A61Q019/08 |
Claims
1. A method for reducing oxidative stress of cells in an individual
in need thereof, the method comprising: topically applying a
composition that comprises at least one active agent selected from
the group consisting of an Undaria extract and a Bidens extract,
and a hydrophilic carrier to a target skin region of the
individual.
2. The method of claim 1 wherein the target skin region is selected
from the group consisting of facial skin, neck skin, breast skin,
shoulder skin, chest skin, leg skin, hand skin, feet skin, and
combinations thereof.
3. The method of claim 1 wherein the active agent is an Undaria
extract, wherein the Undaria extract is present in the composition
in an amount of from about 0.01% (v/v) to about 10% (v/v).
4. The method of claim 3 wherein the Undaria extract is selected
from the group consisting of Undaria crenata, Undaria peterseniana,
Undaria pinnatifida, Undaria undarioides, and combinations
thereof.
5. The method of claim 1 wherein the active agent is a Bidens
extract, wherein the Bidens extract is present in the composition
in an amount of from about 0.01% (v/v) to about 10% (v/v).
6. The method of claim 5 wherein the Bidens extract is selected
from the group consisting of Bidens alba, Bidens amplectens, Bidens
amplissima, Bidens aristosa, Bidens asymmetrica, Bidens aurea,
Bidens beckii, Bidens bidentoides, Bidens bigelovii, Bidens
bipinnata, Bidens biternata, Bidens campylotheca, Bidens cernua,
Bidens cervicata, Bidens chippii, Bidens conjuncta, Bidens connata,
Bidens coronata, Bidens cosmoides, Bidens cynapiifolia, Bidens
discoidea, Bidens eatonii, Bidens ferulifolia, Bidens forbesii,
Bidens frondosa, Bidens gardneri, Bidens hawaiensis, Bidens henryi,
Bidens heterodoxa, Bidens heterosperma, Bidens hillebrandiana,
Bidens hyperborean, Bidens laevis, Bidens lemmonii, Bidens
leptocephala, Bidens leptophylla, Bidens macrocarpa, Bidens mannii,
Bidens mauiensis, Bidens maximowicziana, Bidens menziesii, Bidens
micrantha, Bidens mitis, Bidens molokaiensis, Bidens x multticeps,
Bidens parviflora, Bidens pilosa, Bidens polylepis, Bidens
populifolia, Bidens radiate, Bidens reptans, Bidens sandvicensis,
Bidens schimperi, Bidens simplicifolia, Bidens socorrensis, Bidens
squarrosa, Bidens subalternans, Bidens tenuisecta, Bidens torta,
Bidens trichosperma, Bidens tripartita, Bidens triplinervia, Bidens
valida, Bidens vulgata, Bidens wiebkei, and combinations
thereof.
7. The method of claim 1 wherein the cells are selected from the
group consisting of dermal fibroblasts, epidermal keratinocytes,
and combinations thereof.
8. A method for reducing oxidative stress-induced apoptosis of
cells in an individual in need thereof, the method comprising:
topically applying a composition that comprises at least one active
agent selected from the group consisting of an Undaria extract and
a Bidens extract, and a hydrophilic carrier to a target skin region
of the individual.
9. The method of claim 8 wherein the target skin region is selected
from the group consisting of facial skin, neck skin, breast skin,
shoulder skin, chest skin, leg skin, hand skin, feet skin, and
combinations thereof.
10. The method of claim 8 wherein the active agent is an Undaria
extract, wherein the Undaria extract is present in the composition
in an amount of from about 0.01% (v/v) to about 10% (v/v).
11. The method of claim 10 wherein the Undaria extract is selected
from the group consisting of Undaria crenata, Undaria peterseniana,
Undaria pinnatifida, Undaria undarioides, and combinations
thereof.
12. The method of claim 8 wherein the active agent is a Bidens
extract, wherein the Bidens extract is present in the composition
in an amount of from about 0.01% (v/v) to about 10% (v/v).
13. The method of claim 12 wherein the Bidens extract is selected
from the group consisting of Bidens alba, Bidens amplectens, Bidens
amplissima, Bidens aristosa, Bidens asymmetrica, Bidens aurea,
Bidens beckii, Bidens bidentoides, Bidens bigelovii, Bidens
bipinnata, Bidens biternata, Bidens campylotheca, Bidens cernua,
Bidens cervicata, Bidens chippii, Bidens conjuncta, Bidens connata,
Bidens coronata, Bidens cosmoides, Bidens cynapiifolia, Bidens
discoidea, Bidens eatonii, Bidens ferulifolia, Bidens forbesii,
Bidens frondosa, Bidens gardneri, Bidens hawaiensis, Bidens henryi,
Bidens heterodoxa, Bidens heterosperma, Bidens hillebrandiana,
Bidens hyperborean, Bidens laevis, Bidens lemmonii, Bidens
leptocephala, Bidens leptophylla, Bidens macrocarpa, Bidens mannii,
Bidens mauiensis, Bidens maximowicziana, Bidens menziesii, Bidens
micrantha, Bidens mitis, Bidens molokaiensis, Bidens x multticeps,
Bidens parviflora, Bidens pilosa, Bidens polylepis, Bidens
populifolia, Bidens radiate, Bidens reptans, Bidens sandvicensis,
Bidens schimperi, Bidens simplicifolia, Bidens socorrensis, Bidens
squarrosa, Bidens subalternans, Bidens tenuisecta, Bidens torta,
Bidens trichosperma, Bidens tripartita, Bidens triplinervia, Bidens
valida, Bidens vulgata, Bidens wiebkei, and combinations
thereof.
14. The method of claim 8 wherein the cells are selected from the
group consisting of dermal fibroblasts, epidermal keratinocytes,
and combinations thereof.
15. A method for reducing photodamage of cells in an individual in
need thereof, the method comprising: topically applying a
composition that comprises at least one active agent selected from
the group consisting of an Undaria extract and a Bidens extract,
and a hydrophilic carrier to a target skin region of the
individual.
16. The method of claim 15 wherein the active agent is an Undaria
extract, wherein the Undaria extract is present in the composition
in an amount of from about 0.01% (v/v) to about 10% (v/v).
17. The method of claim 16 wherein the Undaria extract is selected
from the group consisting of Undaria crenata, Undaria peterseniana,
Undaria pinnatifida, Undaria undarioides, and combinations
thereof.
18. The method of claim 15 wherein the active agent is a Bidens
extract, wherein the Bidens extract is present in the composition
in an amount of from about 0.01% (v/v) to about 10% (v/v).
19. The method of claim 18 wherein the Bidens extract is selected
from the group consisting of Bidens alba, Bidens amplectens, Bidens
amplissima, Bidens aristosa, Bidens asymmetrica, Bidens aurea,
Bidens beckii, Bidens bidentoides, Bidens bigelovii, Bidens
bipinnata, Bidens biternata, Bidens campylotheca, Bidens cernua,
Bidens cervicata, Bidens chippii, Bidens conjuncta, Bidens connata,
Bidens coronata, Bidens cosmoides, Bidens cynapiifolia, Bidens
discoidea, Bidens eatonii, Bidens ferulifolia, Bidens forbesii,
Bidens frondosa, Bidens gardneri, Bidens hawaiensis, Bidens henryi,
Bidens heterodoxa, Bidens heterosperma, Bidens hillebrandiana,
Bidens hyperborean, Bidens laevis, Bidens lemmonii, Bidens
leptocephala, Bidens leptophylla, Bidens macrocarpa, Bidens mannii,
Bidens mauiensis, Bidens maximowicziana, Bidens menziesii, Bidens
micrantha, Bidens mitis, Bidens molokaiensis, Bidens x multticeps,
Bidens parviflora, Bidens pilosa, Bidens polylepis, Bidens
populifolia, Bidens radiate, Bidens reptans, Bidens sandvicensis,
Bidens schimperi, Bidens simplicifolia, Bidens socorrensis, Bidens
squarrosa, Bidens subalternans, Bidens tenuisecta, Bidens torta,
Bidens trichosperma, Bidens tripartita, Bidens triplinervia, Bidens
valida, Bidens vulgata, Bidens wiebkei, and combinations
thereof.
20. The method of claim 15 wherein the cells are selected from the
group consisting of dermal fibroblasts, epidermal keratinocytes,
and combinations thereof.
21. The method of claim 1 wherein the composition further includes
beta-sitosterol.
22. The method of claim 21, wherein the beta-sitosterol is present
in the composition in an amount of from about 0.001% by weight of
the composition to about 10.0% by weight of the composition.
23. The method of claim 1, wherein the composition further includes
a vasodilator.
24. The method of claim 23, wherein the vasodilator is selected
from the group consisting of glyceryl trinitrate, resveratrol,
caffeine, ginger extract, ginseng and combinations thereof.
25. The method of claim 8 wherein the composition further includes
beta-sitosterol.
26. The method of claim 25, wherein the beta-sitosterol is present
in the composition in an amount of from about 0.001% by weight of
the composition to about 10.0% by weight of the composition.
27. The method of claim 8, wherein the composition further includes
a vasodilator.
28. The method of claim 27, wherein the vasodilator is selected
from the group consisting of glyceryl trinitrate, resveratrol,
caffeine, ginger extract, ginseng and combinations thereof.
29. The method of claim 15 wherein the composition further includes
beta-sitosterol.
30. The method of claim 29, wherein the beta-sitosterol is present
in the composition in an amount of from about 0.001% by weight of
the composition to about 10.0% by weight of the composition.
31. The method of claim 15, wherein the composition further
includes a vasodilator.
32. The method of claim 31, wherein the-vasodilator is selected
from the group . consisting of glyceryl trinitrate, resveratrol,
caffeine, ginger extract, ginseng and combinations thereof.
Description
BACKGROUND OF THE DISCLOSURE
[0001] The present disclosure relates generally to compositions and
methods for enhancing cellular response to oxidative stress and
photodamage, and in particular, enhancing cellular response in
dermal fibroblasts and epidermal keratinocytes, which can result in
signs of skin aging of the face and body. More particularly, the
present disclosure relates to compositions including an Undaria
extract and/or Bidens extract and methods of topically applying the
compositions for enhancing cellular mitochondrial activity and
antioxidant cellular response to oxidative stress and UV
irradiation, thereby reducing signs of aging of the face and body.
Signs of aging that affect the skin of the face and body include,
for example, wrinkles, fine lines, thinning skin, sagging skin,
skin dryness and skin itchiness.
[0002] With respect to skin aging, a distinction is made between
so-called "intrinsic" and "extrinsic" aging, a decisive factor for
the latter being the exogenous effect, in particular the effect of
ultraviolet (UV) radiation ("photoaging"). Mechanistically,
oxidative stress plays a major role in both intrinsic and extrinsic
aging of the skin since reactive oxygen species (ROS) are generated
in the process of normal cellular metabolism or by physiological
processes and, particularly, are produced by the UVA and UVB
components of UV radiation. If not suppressed upon formation, ROS
have far-reaching effects on the integrity of all cellular
biomolecules such as DNA, proteins and lipids with regard to
UV-induced aging of the skin. Particularly, overproduction of ROS
can lead to lipid peroxidation, damage to biomolecules, and effects
on cellular viability. An immediate consequence of particular
importance in photoaging is the ROS induction of matrix
metalloproteinases that increases degradation of collagen proteins,
creating an imbalance between collagen synthesis and degradation,
leading to skin thinning Thus far, strategies for preventing
photoaging consist of a reduction in UV-exposure, physical
protection from UV-exposure and/or the application of specific
vitamins, such as vitamin C or vitamin E.
[0003] Particularly, a wide variety of creams and lotions exist
that can cosmetically improve the appearance, and sometimes, the
structure of the skin on the face and body. Such compositions often
employ retinoids, hydroxy acids and/or exfoliants to encourage skin
rejuvenation, increase firmness or otherwise cosmetically improve
the skin. Many cosmetic products provide moisture and can enhance
the skin's appearance by plumping the skin using irritants that
cause inflammation. Other cosmetic products are available that
claim to rejuvenate and create a more youthful appearance by
targeting extracellular matrix proteins such as elastin and
collagen, which are produced by fibroblasts to provide skin
strength and resilience.
[0004] Medical procedures, such as dermal injections and
reconstructive surgery, are also available to reduce the signs of
aging. Cosmetic surgery has recently grown in popularity to
aesthetically enhance the appearance of skin. These procedures,
however, are not always desirable options as surgery can be costly,
painful and very invasive.
[0005] While the cosmetic products and medical procedures described
above are suitable for treating aged skin of the face and body,
alternative compositions and methods for improving skin are
desirable. Accordingly, there exists a need to develop alternative
compositions and methods for preventing and/or reducing oxidative
stress that can lead to increased signs of skin aging of the face
and body. It would be highly advantageous if the compositions and
methods could be topically applied such that invasive, painful, and
costly medical procedures could be avoided.
BRIEF DESCRIPTION OF THE DISCLOSURE
[0006] The present disclosure is generally directed to compositions
and methods for enhancing the cellular response to photoaging and
oxidative stress such to reduce the effects of UV irradiation,
oxidative stress and oxidative stress-induced apoptosis of skin
cells, and in particular, dermal fibroblasts and epidermal
keratinocytes. These negative effects on cells can result in
increased signs of skin aging on the face and body.
[0007] In one aspect, the present disclosure is directed to a
method for reducing oxidative stress of cells in an individual in
need thereof. The method includes topically applying a composition
that comprises at least one active agent selected from the group
consisting of an Undaria extract and a Bidens extract, and a
hydrophilic carrier to a target skin region of the individual.
[0008] In another aspect, the present disclosure is directed to a
method for reducing oxidative stress-induced apoptosis of cells in
an individual in need thereof. The method includes topically
applying a composition that comprises at least one active agent
selected from the group consisting of an Undaria extract and a
Bidens extract, and a hydrophilic carrier to a target skin region
of the individual.
[0009] In yet another aspect, the present disclosure is directed to
a method for reducing photodamage of cells in an individual in need
thereof. The method includes topically applying a composition that
comprises at least one active agent selected from the group
consisting of an Undaria extract and a Bidens extract, and a
hydrophilic carrier to a target skin region of the individual.
[0010] In yet another aspect, the present disclosure is directed to
a method for maintaining cellular glutathione concentration of
cells in an individual in need thereof. The method includes
topically applying a composition that comprises at least one active
agent selected from the group consisting of an Undaria extract and
a Bidens extract, and a hydrophilic carrier to a target skin region
of the individual.
BRIEF DESCRIPTION OF THE DRAWINGS
[0011] The disclosure will be better understood, and features,
aspects and advantages other than those set forth above will become
apparent when consideration is given to the following detailed
description thereof. Such detailed description makes reference to
the following drawings, wherein:
[0012] FIG. 1 is a graphical illustration showing viability
following treatment of cells with various concentrations of active
components as discussed in Example 1.
[0013] FIG. 2 is a graph depicting fluorescence emission as an
indicator of oxidative stress as analyzed in Example 2.
[0014] FIGS. 3A-3D are flow cytometer readings of Side Scatter
(SSC) and Forward Scatter (FSC) as measured in Example 3. FIG. 3A
represents control fibroblasts; FIG. 3B represents fibroblasts
treated with H.sub.2O.sub.2 for 1 hour; FIG. 3C represents
fibroblasts treated with UNDARINE.TM. for 12 hours; and FIG. 3D
represents fibroblasts treated with UNDARINE.TM. for 12 hours
followed by exposure to H.sub.2O.sub.2 for 1 hour.
[0015] FIG. 4 is a graph depicting fluorescence emission as an
indicator of oxidative stress as analyzed in Example 4.
[0016] FIG. 5 is a graph depicting fluorescence emission as an
indicator of UV photodamage as analyzed in Example 4.
[0017] FIGS. 6A-6F are flow cytometer readings of Side Scatter
(SSC) and Forward Scatter (FSC) as measured in Example 5. FIG. 6A
represents control fibroblasts; FIG. 6B represents fibroblasts
treated with H.sub.2O.sub.2 for 1 hour; FIG. 6C represents
fibroblasts treated with ECOBIDENS.TM. for 12 hours followed by
exposure to H.sub.2O.sub.2 for 1 hour; FIG. 6D represents
fibroblasts treated with ECOBIDENS.TM. for 12 hours; FIG. 6E
represents fibroblasts exposed to UV irradiation; and FIG. 6F
represents fibroblasts treated with ECOBIDENS.TM. for 12 hours
followed by exposure to UV irradiation.
[0018] FIG. 7 is a graph depicting decreased cellular GSH
concentration due to UV irradiation or H.sub.2O.sub.2-induced
oxidative stress as analyzed in Example 6.
[0019] FIG. 8 is a graph depicting activity of ECOBIDENS.TM. on the
concentration of H.sub.2O.sub.2 as analyzed in Example 7.
[0020] FIG. 9 is a graph depicting fluorescence emission as an
indicator of H.sub.2O.sub.2-induced oxidative stress as analyzed in
Example 8.
[0021] FIG. 10 is a graph depicting fluorescence emission as an
indicator of UV exposure as analyzed in Example 8.
[0022] While the disclosure is susceptible to various modifications
and alternative forms, specific embodiments thereof have been shown
by way of example in the drawings and are herein described below in
detail. It should be understood, however, that the description of
specific embodiments is not intended to limit the disclosure to
cover all modifications, equivalents and alternatives falling
within the spirit and scope of the disclosure as defined by the
appended claims.
DETAILED DESCRIPTION OF THE DISCLOSURE
[0023] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which the disclosure belongs. Although
any methods and materials similar to or equivalent to those
described herein can be used in the practice or testing of the
present disclosure, the preferred methods and materials are
described below.
[0024] In accordance with the present disclosure, compositions and
methods have been discovered that surprisingly allow for reducing
the signs of skin aging. The methods of the present disclosure have
a broad and significant impact, as they protect cells from UV
exposure and excessive production of ROS by stimulating the
cellular antioxidant response (e.g., increase in reduced
gluthatione concentration), thereby resulting in a more youthful
appearance of aged skin of the face and body. More particularly, in
the dermis, fibroblasts produce collagen and elastin, which provide
skin strength and resilience. Photodamage and ROS-induced oxidative
stress are among the main causes of fibroblast degeneration and
death; thereby, accelerating the process of skin aging and
exacerbating skin aging symptoms such as skin thinning, sagging,
fragility, and loss of skin resiliency.
[0025] As used herein, "body" refers to an individual's entire
body, and particularly, includes the regions of the face (including
forehead, cheeks, chin, and eyelids), neck, shoulders, breast,
chest, legs, hands, feet and vulvar area including vulvar skin, for
example, the vulva, labia, labia majora, labia minor, mons pubis,
vulval vestibule, and combinations thereof
[0026] As used herein, "apoptosis" refers to the process of
programmed cell death that may occur in multicellular organisms.
Particularly, a cell initiates intracellular apoptotic signaling in
response to a stress (e.g., UV irradiation, oxidative stress
resulting from ROS overproduction, etc.).
COMPOSITIONS
Active Ingredients/Extracts
[0027] The present disclosure has unexpectedly found that
particular extracts can be topically applied, alone or in
combination, to an individual's skin as active ingredients in
compositions to reduce the signs of skin aging. Particularly, while
the active ingredients of the compositions have been previously
known to stimulate collagen and elastin, it has now been
unexpectedly found that these active ingredients are effective at
protecting the skin from factors that cause extrinsic skin aging,
and in particular, to protect the dermal layer from exposure to UV
irradiation and ROS-induced damage. It is believed that this
protective effect is due to the active agents enhancing cellular
defense mechanisms, and in particular, cellular mitochondrial
activity and antioxidant cellular response. This provides the user
with plumper, fuller, more resilient skin and a more youthful,
smoother skin appearance.
Undaria Extract
[0028] In one aspect, the present disclosure is directed to
compositions for reducing the signs of skin aging of the face and
body; the compositions generally include an Undaria extract as the
active ingredient. As used herein, "extract" refers to the active
solid components from the Undaria plant.
[0029] An Undaria extract is an extract obtained from Undaria, a
genus of brown alga that includes U crenata, U peter seniana, U
pinnatifida and U undarioides. Undaria extracts are known to
stimulate collagen and elastin, and have previously been described
in vendor literature to increase 14 genes linked to collagen and
elastin production. As disclosed herein, the Undaria extracts used
in the compositions of the present disclosure have further
surprisingly been found to protect cells, and in particular, dermal
fibroblasts and epidermal keratinocytes, from photodamage and
oxidative stress.
[0030] As used herein, Undaria extract, refers to a liquid extract
from the Undaria plant. In particularly suitable embodiments, the
Undaria extract has not been further supplemented/standardized for
fucoidan content. Particularly, the Undaria extracts in these
suitable embodiments are extracts obtained by soaking or passing
the Undaria algae plants in/over water with no further chemical
treatment. The soaking/passing over can be accomplished with any
method known in the extraction arts, such as, for example,
lixiviation, infusion steeping, percolation, extraction, and the
like, and combinations thereof. In some embodiments, the Undaria
plant could be mascerated to further obtain the extract.
Additionally, the water used in these processes could be heated to
obtain the Undaria extract.
[0031] One particularly suitable method for obtaining the Undaria
extract is lixiviation of fertile bases from lyophilized Undaria
algae plants. As used herein, "lixiviation" refers to a technique
where water is slowly passed over a solid (here, Undaria) in a
layer of varying depth to extract soluble material. The general
technique is described in FR 2693917, which is incorporated herein
to the extent it is consistent herewith, and is performed at room
temperature. After removal of the extract, in some embodiments, the
extract is further diluted with diluents including water, glycerin,
propanediol, butylene glycol, propylene glycol and combinations
thereof.
[0032] By contrast, Undaria extracts that are
supplemented/standardized for fucoidan content typically are
obtained by an extraction procedure utilizing an acid/water mixture
having a pH of between 0 and 2, preferably between about 0 and 1,
at temperatures between about 0 and 30.degree. C., preferably
between about 15 and 25.degree. C.
[0033] Suitable Undaria extracts can be obtained from commercially
available sources (e.g., Barnet Products Corp., Englewood Cliffs,
N.J.; Changsha Organic Herb Inc., Hunan, China). A particularly
suitable commercially available formulation including Undaria
extract can be, for example, UNDARINE.TM., which is a formulation
including glycerin, water and U. pinnatifida extract (commercially
available from Barnet Products Corp., Englewood Cliffs, N.J.).
[0034] Another suitable commercially available Undaria extract can
be, for example, the U. pinnatifida extract WAKAMINE.RTM. (INCI:
water (and) Undaria pinnatifida extract), commercially available
from SOLIANCE (France), which has previously been used to lighten
skin and prevent age-spots.
[0035] Suitable amounts of an Undaria extract in the composition
can be from about 0.01% (w/w) to about 10% (w/w), from about 0.05%
(w/w) to about 7.5% (w/w), from about 0.1% (w/w) to about 5% (w/w),
and from about 1% (w/w) to about 5% (w/w). As used herein "w/w"
refers to the amount of the component "by weight of the
composition".
Bidens Extract
[0036] In another aspect, the present disclosure is directed to a
composition including a Bidens extract as the active agent. As used
herein, "extract" refers to the active solid components from the
Bidens plant. Bidens is a genus of plants in the Asteraceae family
that includes many species members. Bidens is known to increase
collagen and elastin production.
[0037] Suitable Bidens extracts include extracts of Bidens alba,
Bidens amplectens, Bidens amplissima, Bidens aristosa, Bidens
asymmetrica, Bidens aurea, Bidens beckii, Bidens bidentoides,
Bidens bigelovii, Bidens bipinnata, Bidens biternata, Bidens
campylotheca, Bidens cernua, Bidens cervicata, Bidens chippii,
Bidens conjuncta, Bidens connata, Bidens coronata, Bidens
cosmoides, Bidens cynapiifolia, Bidens discoidea, Bidens eatonii,
Bidens ferulifolia, Bidens forbesii, Bidens frondosa, Bidens
gardneri, Bidens hawaiensis, Bidens henryi, Bidens heterodoxa,
Bidens heterosperma, Bidens hillebrandiana, Bidens hyperborean,
Bidens laevis, Bidens lemmonii, Bidens leptocephala, Bidens
leptophylla, Bidens macrocarpa, Bidens mannii, Bidens mauiensis,
Bidens maximowicziana, Bidens menziesii, Bidens micrantha, Bidens
mitis, Bidens molokaiensis, Bidens x multticeps, Bidens parviflora,
Bidens pilosa, Bidens polylepis, Bidens populifolia, Bidens
radiate, Bidens reptans, Bidens sandvicensis, Bidens schimperi,
Bidens simplicifolia, Bidens socorrensis, Bidens squarrosa, Bidens
subalternans, Bidens tenuisecta, Bidens torta, Bidens trichosperma,
Bidens triparfita, Bidens triplinervia, Bidens valida, Bidens
vulgata, Bidens wiebkei, and combinations thereof.
[0038] Particularly suitable Bidens extracts can be, for example,
Bidens pilosa extract, Bidens bipinnata extract, and Bidens
tripartita extract. Suitable Bidens extract can be obtained from
commercially available sources (e.g., Chemyunion Quimica Ltd., Sao
Paulo, Brazil; and Carrubba Inc., Milford, Conn.). Suitable Bidens
extracts can also include Bidens extract blends with hydrophilic
carriers such as for example, water, glycerin, propanediol,
butylene glycol, propylene glycol and combinations thereof and
Bidens extract blends with hydrophobic carriers such as for example
natural oils, synthetic oils and combinations thereof. A
particularly suitable Bidens extract can be, for example,
ECOBIDENS.TM. (commercially available from Chemyunion Quimica Ltd.,
Sao Paulo, Brazil). ECOBIDENS.TM. is a glycerin extract of Bidens
pilosa L. Other commercially available Bidens extracts include, for
example, Water Agrimony Extract H. G., a B. tripartita extract from
Provital Group (Spain); Bidens Extract M9983-WS, a B. pilosa
extract, and Bidens bipinnata Extract N0061-WS, a B. bipinnata
extract, both available from Carrubba Inc. (Milford, Conn.).
[0039] Suitable amounts of Bidens extract in the topical
compositions can be from about 0.01% (w/w) to about 10% (w/w), from
about 0.05% (w/w) to about 7.5% (w/w), from about 0.1% (w/w) to
about 5% (w/w), and from about 1% (w/w) to about 5% (w/w).
Undaria Extract and Bidens Extract
[0040] In some particularly suitable aspects, the present
disclosure is directed to a composition including combination of
the active agents of an Undaria extract and a Bidens extract.
[0041] When used in a combination, suitable amounts of the Undaria
extract in the composition can be from about 0.01% (w/w) to about
20% (w/w), from about 0.05% (w/w) to about 15% (w/w), and from
about 0.1% (w/w) to about 10% (w/w). Suitable amounts of Bidens
extract in the composition can be from about 0.01% (w/w) to about
20% (w/w), from about 0.05% (w/w) to about 15% (w/w), and from
about 0.1% (w/w) to about 10% (w/w).
Additional Optional Ingredients
[0042] The compositions described herein can further include
additional ingredients and optional ingredients.
[0043] Generally, the compositions include a carrier with the
active ingredient(s).
[0044] In one embodiment, for example, the composition includes the
active ingredient(s) and a hydrophilic carrier, a hydrophilic
thickener, and/or a penetration enhancer. Suitable hydrophilic
carriers can be, for example, water, alcohols, glycerin, glycerin
derivatives, glycols, water-soluble emollients, and combinations
thereof. Suitable examples of alcohols could include, but are not
to be limited to, ethanol and isopropyl alcohol. Suitable examples
of glycerin derivatives could include, but are not to be limited
to, PEG-7 glyceryl cocoate. Suitable glycols could include, but are
not to be limited to, propylene glycol, butylene glycol, pentylene
glycol, ethoxydiglycol, dipropylene glycol, propanediol, and PEG-8.
Suitable examples of water-soluble emollients could include, but
are not to be limited to, PEG-6 Caprylic Capric Glycerides,
Hydrolyzed Jojoba Esters, and PEG-10 Sunflower Glycerides.
[0045] In particularly suitable embodiments, the topical
compositions are liquid compositions desirably containing water as
the carrier. Suitable amounts of water can be from about 0.1% by
weight of the composition to about 99.9% by weight of the
composition. More typically, the amount of water can be from about
40% by weight of the composition to about 99.9% by weight of the
composition. Preferably, the amount of water can be from about 60%
by weight of the composition to about 99.9% by weight of the
composition.
[0046] In another embodiment, the composition includes the active
ingredient(s) and a hydrophobic carrier. Suitable hydrophobic
carriers can be, for example, natural oils, synthetic oils and
combinations thereof
[0047] The topical compositions described herein can further
include a skin penetrating enhancer or a mixture of skin
penetration enhancers. Examples of suitable skin penetration
enhancers include sulfoxides, alcohols, fatty acids, fatty acid
esters, polyols, amides, surfactants, terpenes, alkanones, and
organic acids, among others. Specific examples of suitable
sulfoxides include dimethylsulfoxide (DMSO) and
decylmethylsulfoxide, among others. Suitable alcohols include
alkanols such as ethanol, propanol, butanol, pentanol, hexanol,
octanol, n-octanol, nonanol, decanol, 2-butanol, 2-pentanol, and
benzyl alcohol; fatty alcohols, such as caprylic alcohol, decyl
alcohol, lauryl alcohol, 2-lauryl alcohol, myristyl alcohol, cetyl
alcohol, stearyl alcohol, oleyl alcohol, linoleyl alcohol, and
linolenyl alcohol; and isopropyl alcohol. Examples of suitable
fatty acids include linear fatty acids such as valeric acid,
heptanoic acid, pelagonic acid, caproic acid, capric acid, lauric
acid, myristic acid, stearic acid, oleic acid, and caprylic acid;
and branched fatty acids, such as isovaleric acid, neopentanoic
acid, neoheptanoic acid, neononanoic acid, trimethyl hexanoic acid,
neodecanoic acid, and isostearic acid. Examples of suitable fatty
acid esters include aliphatic fatty acid esters such as isopropyl
n-butyrate, isopropyl n-hexanoate, isopropyl n-decanoate, isopropyl
myristate, isopropyl palmitate, and octyldodecyl myristate; alkyl
fatty acid esters such as ethyl acetate, butyl acetate, methyl
acetate, methylvalerate, methylpropionate, diethyl sebacate, and
ethyl oleate; and diisopropyl adipate and dimethyl isosorbide.
Examples of suitable polyols include propylene glycol, butylene
glycol, polyethylene glycol, ethylene glycol, diethylene glycol,
triethylene glycol, dipropylene glycol, ethoxydiglycol, pentylene
glycol, glycerol, propanediol, butanediol, pentanediol,
hexanetriol, and glycerin. Examples of suitable amides include
urea, dimethylacetamide, diethyltoluamide, dimethylformamide (DMF),
dimethyloctamide, dimethyldecamide, biodegradable cyclic urea
(e.g., 1-alkyl-4-imidazoline-2-one), pyrrolidone derivatives,
biodegradable pyrrolidone derivatives (e.g., fatty acid esters of
N-(2-hydroxyethyl)-2-pyrrolidone), cyclic amides,
hexamethylenelauramide and its derivatives, diethanolamine, and
triethanolamine. Examples of pyrrolidone derivatives include
1-methyl-2-pyrrolidone, 2-pyrrolidone, 1-lauryl-2-pyrrolidone,
1-methyl-4-carboxy-2-pyrrolidone, 1-hexyl-4-carboxy-2-pyrrolidone,
1-lauryl-4-carboxy-2-pyrrolidone,
1-methyl-4-methoxycarbonyl-2-pyrrolidone,
1-hexyl-4-methoxycarbonyl-2-pyrrolidone,
1-lauryl-4-methoxycarbonyl-2-pyrrolidone, N-cyclohexylpyrrolidone,
N-dimethylaminopropylpyrrolidone, N-cocoalkypyrrolidone,
N-tallowalkylpyrrolidone, and N-methylpyrrolidone. Examples of
cyclic amides include 1-dodecylazacycloheptane-2-one (e.g.,
Azone.RTM.), 1-geranylazacycloheptan-2-one, 1-farnesylaz
acycloheptan-2-one, 1-geranylgeranylazacycloheptan-2-one,
1-(3,7-dimethyloctyl)azacycloheptan-2-one,
1-(3,7,11-trimethyldodecyl)azacyclohaptane-2-one,
1-geranylazacyclohexane-2-one, 1-geranylazacyclopentan-2,5-dione,
and 1-farnesylazacyclopentan-2-one.
[0048] Typically, the compositions of the present disclosure
include from about 0.01% (by weight of the composition) to about
25% (by weight of the composition) of a skin penetration enhancer,
including from about 1% (by weight of the composition) to about 15%
(by weight of the composition) of a skin penetration enhancer, and
including from about 2% (by weight of the composition) to about 10%
(by weight of the composition) of a skin penetration enhancer.
[0049] Optionally, the Undaria and/or Bidens-containing topical
compositions may be formulated with a polar co-solvent to increase
the permeability of the Undaria and Bidens into the skin.
Preferably, the polar co-solvent is fully miscible in the
composition, and has a high affinity for the intercellular spaces
in the stratum corneum. Without wishing to be bound by any
particular theory, it is believed that polar co-solvents with such
characteristics are driven by osmosis into the intercellular spaces
in the stratum corneum, causing the stratum corneum to swell. In
such a swollen state, the intercellular spaces are more liquid-like
and disordered, which enables the Undaria and/or Bidens extracts to
more easily diffuse through the stratum corneum.
[0050] Examples of suitable polar co-solvents for inclusion in the
compositions of the present disclosure include glycerin,
propanediol, ethanol, propylene glycol, butanol, isopropanol,
propanol, dimethyl isosorbide, butylene glycol, polyethylene
glycol, dipropylene glycol, ethoxydiglycol, pentylene glycol, and
combinations thereof
[0051] Typically, the compositions of the present disclosure
include from about 1% (by weight of the composition) to about 99%
(by weight of the composition) of a polar co-solvent, including
from about 1% (by weight of the composition) to about 75% (by
weight of the composition) of a polar co-solvent, including from
about 1% (by weight of the composition) to about 50% (by weight of
the composition) of a polar co-solvent, including from about 1.5%
(by weight of the composition) to about 25% (by weight of the
composition) of a polar co-solvent, including from about 2% (by
weight of the composition) to about 15% (by weight of the
composition) of a polar co-solvent, and including from about 2.5%
(by weight of the composition) to about 10% (by weight of the
composition) of a polar co-solvent.
[0052] In some embodiments, the compositions can further include
beta-sitosterol. Suitable amounts of beta-sitosterol can be from
about 0.001% by weight of the composition to about 10.0% by weight
of the composition. More typically, the amount of beta-sitosterol
can be from about 0.005% by weight of the composition to about 7.5%
by weight of the composition. Even more suitable, the amount of
beta-sitosterol can be from about 0.01% by weight of the
composition to about 5.0% by weight of the composition.
[0053] The composition can further include other known collagen,
elastin, and extracellular matrix-stimulating ingredients. Collagen
is a protein found in the connective tissue of the skin and other
tissues of the body. Suitable collagen enhancers can be, for
example, vitamins such as ascorbic acid and derivatives thereof,
peptides such as palmitoyl tripeptide-5, botanical extracts such as
pomegranate or mushroom, and minerals such as hematite.
[0054] Elastin is a protein found in the connective tissue of the
skin and other tissues of the body. Suitable elastin enhancers can
be, for example, vitamins such as ascorbic acid and derivatives
thereof, peptides such palmitoyl hexapeptide-12, botanical extracts
such as kudzu, horsetail, rice, dill and rosemary, and minerals
such as zinc and copper.
[0055] The compositions can further include a vasodilator.
Vasodilators can increase the blood flow within the skin. Suitable
vasodilators can be, for example, glyceryl trinitrate, resveratrol,
caffeine, ginger extract, ginseng and other botanical extracts such
as, for example, hawthorn, mint, ivy, coffee and tea.
[0056] The compositions can further include a skin soothing agent.
As used herein, "skin soothing agent" refers to compounds that
reduce or prevent skin irritation. Skin irritation can result from
loss of moisture, a change in pH, sweat, contact dermatitis from
perfumes, powders, laundry detergent from clothing, and other
compounds. Skin soothing agents can reduce irritation by
neutralizing an irritant, down-regulating inflammatory cascades in
the skin, and/or providing a protective layer on the skin. Suitable
skin soothing agents can be, for example, botanical extracts such
as calendula, chamomile, aloe, comfrey, coneflower; active
materials such as allantoin, bisabolol, panthenol, beta-glucan,
colloidal oatmeal, and ingredient blends such as SYMCALMIN (INCI:
butylene glycol, pentylene glycol, hydroxyphenyl propamidobenzoic
acid; commercially available from Symrise (Holzmiden, Germany) and
SEPICALM (INCI: sodium palmitoyl proline, nymphaea alba flower
extract; commercially available from Seppic (Fairfield, N.J.).
[0057] The compositions can further include a humectant. Humectants
can elevate the hydration of the skin, in particular the epidermis
and the dermis. Suitable humectants can be, for example, glycerol,
glycerin, lactic acid, urea, aloe vera, betaine, hyaluronic acid,
propanediol, propylene glycol, butylene glycol, and combinations
thereof
[0058] The compositions can further include an emulsifier, and in
particular, an emulsifier that creates liquid crystalline networks
or liposomal networks. Suitable non-limiting exemplary emulsifiers
include, for example, OLIVEMO 1000 (INCI: Cetearyl Olivate (and)
Sorbitan Olivate; commercially available from HallStar Company
(Chicago, Ill.)), Arlacel.TM. LC (INCI: Sorbitan Stearate (and)
Sorbityl Laurate; commercially available from Croda (Edison, N.J.),
CRYSTALCAST.RTM. MM (INCI: Beta Sitosterol (and) Sucrose Stearate
(and) Sucrose Distearate (and) Cetyl Alcohol (and) Stearyl Alcohol;
commercially available from MMP Inc. (South Plainfield, N.J.),
UNIOX CRISTAL (INCI: Cetearyl Alcohol (and) Polysorbate 60 (and)
Cetearyl Glucoside; commercially available from Chemyunion (Sao
Paulo, Brazil). Other suitable emulsifiers include lecithin,
hydrogenated lecithin, lysolecithin, phosphatidylcholine,
phospholipids, and combinations thereof.
[0059] The compositions can further include a preservative to
preserve the stability. Preservatives can also prevent the growth
of microbial organisms in the compositions. Suitable preservatives
are known in the art, and include, for example, methylparaben,
phenoxyethanol, capryl glycol, glyceryl caprylate, benzoic acid,
sorbic acid, gallic acid, propylparaben and combinations
thereof.
[0060] The compositions can further include a pH adjuster to
control/maintain the pH of the composition within the range of skin
pH. A suitable pH range of the composition can be from about 3.5 to
about 6.
[0061] The compositions can further include fragrances, scents,
dyes, surfactants, rheology modifiers, film formers and other
components known to be useful in personal care formulations.
Methods of Use
[0062] In another aspect, the present disclosure is directed to
methods of using the compositions to reduce skin aging of an
individual's face and body.
[0063] Thus, in one aspect, the present disclosure is directed to a
method for reducing oxidative stress of cells, and in particular
dermal fibroblasts and epidermal keratinocytes, thereby reducing
signs of skin aging of the face and body in an individual in need
thereof. As used herein, "skin aging" refers to increased skin
wrinkling, increased appearance of fine lines, thinning skin,
sagging skin, skin dryness, and skin itchiness. Oxidative stress of
cells alters the equilibrium between cellular death and cellular
proliferation, and further, can impair tissue regeneration.
Typically, oxidative stress is a result of the overproduction of
reactive oxygen species (ROS), which causes damage to the inner and
outer mitochondrial membranes and opens the mitochondrial
permeability transition pores, thereby inducing apoptosis. The
method includes topically applying a composition including an
Undaria extract to a target skin region of the individual. The
target skin region can be, for example, facial skin, neck skin,
breast skin, shoulder skin, chest skin, leg skin, hand skin, feet
skin, and combinations thereof. The target skin region can also be,
for example, vulvar skin, for example, the vulva, labia, labia
majora, labia minor, mons pubis, vulval vestibule and combinations
thereof
[0064] In another aspect, the present disclosure is directed to a
method for reducing oxidative stress-induced apoptosis of cells,
and in particular dermal fibroblasts and epidermal keratinocytes,
in an individual in need thereof. The method includes topically
applying a composition including an Undaria extract to a target
skin region of the individual. The target skin region can be, for
example, facial skin, neck skin, breast skin, shoulder skin, chest
skin, leg skin, hand skin, feet skin, and combinations thereof. The
target skin region can also be, for example, vulvar skin, for
example, the vulva, labia, labia majora, labia minor, mons pubis,
vulval vestibule and combinations thereof
[0065] In yet another aspect, the present disclosure is directed to
a method for reducing photodamage of cells, and in particular
dermal fibroblasts and epidermal keratinocytes, in an individual in
need thereof. As used herein, "photodamage" refers to the damage
caused to skin, and in particular, dermal fibroblasts, as a result
of exposure to UV irradiation. The method includes topically
applying a composition including an Undaria extract to a target
skin region of the individual. The target skin region can be, for
example, facial skin, neck skin, breast skin, shoulder skin, chest
skin, leg skin, hand skin, feet skin, and combinations thereof. The
target skin region can also be, for example, vulvar skin, for
example, the vulva, labia, labia majora, labia minor, mons pubis,
vulval vestibule and combinations thereof.
[0066] In yet another aspect, the present disclosure is directed to
a method for maintaining the cellular reduced glutathione
concentration of cells, and in particular dermal fibroblasts and
epidermal keratinocytes, in an individual in need thereof.
Particularly, the methods are directed to maintaining cellular
reduced glutathione concentration of dermal fibroblasts under
oxidative stress and/or UV irradiation conditions. The methods
include topically applying a composition that comprises an Undaria
extract and a hydrophilic carrier to a target skin region of the
individual. The target skin region can be, for example, facial
skin, neck skin, breast skin, shoulder skin, chest skin, leg skin,
hand skin, feet skin, vulvar skin, for example, the vulva, labia,
labia majora, labia minor, mons pubis, and vulval vestibule, and
combinations thereof.
[0067] In yet other embodiments of the present disclosure, the
methods include topically applying a composition including a Bidens
extract to a target skin region of the individual to reduce
oxidative stress of cells, and in particular dermal fibroblasts and
epidermal keratinocytes. The target skin region can be, for
example, facial skin, neck skin, breast skin, shoulder skin, chest
skin, leg skin, hand skin, feet skin, and combinations thereof. The
target skin region can also be, for example, vulvar skin, for
example, the vulva, labia, labia majora, labia minor, mons pubis,
vulval vestibule and combinations thereof
[0068] In another aspect, the present disclosure is directed to a
method for reducing oxidative stress-induced apoptosis of cells,
and in particular dermal fibroblasts and epidermal keratinocytes,
in an individual in need thereof. The method includes topically
applying a composition including a Bidens extract to a target skin
region of the individual. The target skin region can be, for
example, facial skin, neck skin, breast skin, shoulder skin, chest
skin, leg skin, hand skin, feet skin, and combinations thereof. The
target skin region can also be, for example, vulvar skin, for
example, the vulva, labia, labia majora, labia minor, mons pubis,
vulval vestibule and combinations thereof
[0069] In yet another aspect, the present disclosure is directed to
a method for reducing photodamage of cells, and in particular
dermal fibroblasts and epidermal keratinocytes, in an individual in
need thereof. The method includes topically applying a composition
including a Bidens extract to a target skin region of the
individual. The target skin region can be, for example, facial
skin, neck skin, breast skin, shoulder skin, chest skin, leg skin,
hand skin, feet skin, and combinations thereof. The target skin
region can also be, for example, vulvar skin, for example, the
vulva, labia, labia majora, labia minor, mons pubis, vulval
vestibule and combinations thereof
[0070] In yet another aspect, the present disclosure is directed to
a method for maintaining the cellular reduced glutathione
concentration of cells, and in particular dermal fibroblasts and
epidermal keratinocytes, in an individual in need thereof.
Particularly, the methods are directed to maintaining cellular
reduced glutathione concentration of dermal fibroblasts under
oxidative stress and/or UV irradiation conditions. The methods
include topically applying a composition that comprises a Bidens
extract and a hydrophilic carrier to a target skin region of the
individual. The target skin region can be, for example, facial
skin, neck skin, breast skin, shoulder skin, chest skin, leg skin,
hand skin, feet skin, vulvar skin, for example, the vulva, labia,
labia majora, labia minor, mons pubis, and vulval vestibule, and
combinations thereof.
[0071] As used herein, an "individual in need" refers to an
individual showing increased visible signs of skin aging such as,
for example, wrinkles, fine lines, thinning skin, sagging skin,
skin dryness, skin itchiness, skin fragility, skin tears, loss in
skin tone, loss in skin fullness, loss in skin plumpness, and
combinations thereof, due to extrinsic conditions, and in
particular, oxidative stress and/or photodamage of cells, and in
particular, dermal fibroblasts and epidermal keratinocytes. In some
embodiments, the individual in need is an individual in which the
cellular glutathione concentration of dermal fibroblasts has
decreased as a result of oxidative stress and/or UV irradiation. As
such, in some embodiments, the methods disclosed herein are
directed to a subset of the general population such that, in these
embodiments, not all of the general population may benefit from the
methods. Based on the foregoing, because some of the method
embodiments of the present disclosure are directed to specific
subsets or subclasses of identified individuals (that is, the
subset or subclass of individuals "in need" of assistance in
addressing one or more specific conditions noted herein), not all
individuals will fall within the subset or subclass of individuals
as described herein.
[0072] As used herein, the term "individual" refers to a male human
or a female human. In certain embodiments, the individual is a
postmenopausal female human.
[0073] The compositions used in the methods described herein can
further include additional ingredients as described herein and
other components known to be useful in personal care
formulations.
[0074] The composition can be applied to the target skin region by
any suitable delivery vehicle. For example, the composition can be
applied as a lotion, as a wash, as a gel, as a salve, as an
ointment, as a cream, as a solid stick and as a foam. Additionally,
the composition can be applied with a wipe, with mitts and gloves,
using an aerosol dispenser, using a pump spray, using a trigger
spray and using a squeeze bottle.
[0075] The compositions can be applied daily, every other day,
every couple of days, every week, every month, and every year, as
desired. The compositions can be applied multiple times per day,
multiple times per week and/or multiple times per month.
[0076] In some embodiments, the compositions of the present
disclosure can be used with additional skin care compositions as
part of a skin care regimen. For example, in facial treatment and
care, users typically use multiple products for cleansing, toning,
and treating the skin of the face. Accordingly, the first product
comprises a first composition typically capable of providing a
first benefit to a user, and the second product comprises a second
composition typically capable of providing a second benefit to a
user. In the present disclosure, it should be understood that at
least one of the products of the regime includes the topical
composition of the present disclosure, thereby providing the
benefit of enhancing the cellular response to oxidative stress
and/or photodamage. In some embodiments, it should be understood by
one skilled in the art, that while the first product and second
product can independently provide any benefit known in the art of
the particular care regimen, in each particular multi-product care
regimen, the first product and second product may include different
compositions and thus, provide different benefits to the user.
[0077] Furthermore, as with the first product and second product of
the care regimen, if more than two products are used in the
multi-product care regimen, such as third, fourth, and/or fifth
products (and more products if more than five products are
desired), it should be recognized that the additional products
should include active ingredients, each independently being capable
of providing additional benefits to a user.
[0078] In an alternative embodiment, the multi-product care regime
can include more than two products and can be configured such to
provide a multiple day regimen. Without being limiting, in one
example, the multi-product care regimen provides skin care for
multiple days and, as such, a first product includes, for example,
a cleansing composition, a second product includes the anti-skin
aging composition of the present disclosure, a third product
includes the same cleansing composition as the first product, and a
fourth product includes the same anti-skin aging composition as the
second product.
[0079] Without being limiting, examples of additional compositions
providing skin care benefits in addition to the anti-skin aging
composition of the present disclosure can include compositions for
cleansing, toning, treating, moisturizing, protecting, finishing,
and the like.
[0080] When the additional composition is a cleansing composition,
the cleansing composition may be in any form known in the art, such
as, for example, hand soaps, body soaps, body washes, shampoos,
surface cleaners, dish soaps, facial cleansers, hand washes, and
the like. These types of cleansing compositions typically include
at least one foaming agent, such as a surfactant. Although
discussed herein primarily in terms of a surfactant, it should be
understood that the cleansing compositions may comprise other
cleansing agents, and need not comprise a surfactant. For example,
in certain embodiments, the compositions may comprise a thickener,
a swellable clay, a foaming agent (which may or may not comprise a
surfactant (e.g., ethoxylated skin conditioning agents,
solubilizers, and derivatized silicone polymers)), and optionally a
solvent or other carrier. Examples of such compositions include,
for example, lotions, creams, anti-microbial compositions, and the
like.
[0081] Suitable surfactants for use in the cleansing composition
include anionic surfactants, amphoteric surfactants, cationic
surfactants, zwitterionic surfactants, non-ionic surfactants, and
combinations thereof
[0082] Suitable anionic surfactants include, for example, alkyl
sulfates, alkyl ether sulfates, alkyl aryl sulfonates, alpha-olefin
sulfonates, alkali metal or ammonium salts of alkyl sulfates,
alkali metal or ammonium salts of alkyl ether sulfates, alkyl
phosphates, silicone phosphates, alkyl glyceryl sulfonates, alkyl
sulfosuccinates, alkyl taurates, acyl taurates, alkyl sarcosinates,
acyl sarcosinates, sulfoacetates, alkyl phosphate esters, mono
alkyl succinates, monoalkyl maleates, sulphoacetates, acyl
isethionates, alkyl carboxylates, phosphate esters,
sulphosuccinates (e.g., sodium dioctylsulphosuccinate), and
combinations thereof. Specific examples of anionic surfactants
include sodium lauryl sulphate, sodium lauryl ether sulphate,
ammonium lauryl sulphosuccinate, ammonium lauryl sulphate, ammonium
lauryl ether sulphate, sodium dodecylbenzene sulphonate,
triethanolamine dodecylbenzene sulphonate, sodium cocoyl
isethionate, sodium lauryl sulphate, sodium N-lauryl sarcosinate,
and combinations thereof.
[0083] Suitable cationic surfactants include, for example, alkyl
ammonium salts, polymeric ammonium salts, alkyl pyridinium salts,
aryl ammonium salts, alkyl aryl ammonium salts, silicone quaternary
ammonium compounds, and combinations thereof Specific examples of
cationic surfactants include behenyltrimonium chloride,
stearlkonium chloride, distearalkonium chloride, chlorohexidine
diglutamate, polyhexamethylene biguanide (PHMB), cetyl pyridinium
chloride, benzammonium chloride, benzalkonium chloride, and
combinations thereof
[0084] Suitable amphoteric surfactants include, for example,
betaines, alkylamido betaines, sulfobetaines, N-alkyl betaines,
sultaines, amphoacetates, amophodiacetates, imidazoline
carboxylates, sarcosinates, acylamphoglycinates, such as
cocamphocarboxyglycinates and acylamphopropionates, and
combinations thereof. Specific examples of amphoteric surfactants
include cocamidopropyl betaine, lauramidopropyl betaine,
meadowfoamamidopropyl betaine, sodium cocoyl sarcosinate, sodium
cocamphoacetate, disodium cocoamphodiacetate, ammonium cocoyl
sarcosinate, sodium cocoamphopropionate, and combinations
thereof
[0085] Suitable zwitterionic surfactants include, for example,
alkyl amine oxides, silicone amine oxides, and combinations
thereof. Specific examples of suitable zwitterionic surfactants
include, for example,
4-[N,N-di(2-hydroxyethyl)-N-octadecylammonio]-butane-1-carboxylate,
5-[S-3
-hydroxypropyl-S-hexadecylsulfonio]-3-hydroxypentane-1-sulfate,
3-[P,P-diethyl-P-3,6,9-trioxatetradexopcylphosphonio]-2-hydroxypropane-1--
phosphate,
3-[N,N-dipropyl-N-3-dodecoxy-2-hydroxypropylammonio]-propane-1--
phosphonate,
3-(N,N-dimethyl-N-hexadecylammonio)propane-1-sulfonate,
3-(N,N-dimethyl-N-hexadecylammonio)-2-hydroxypropane-1-sulfonate,
4-[N,N-di(2-hydroxyethyl)-N-(2-hydroxydodecyl)ammonio]-butane-1-carboxyla-
te,
3-[S-ethyl-S-(3-dodecoxy-2-hydroxypropyl)sulfonio]-propane-1-phosphate-
, 3-[P,P-dimethyl-P-dodecylphosphonio]-propane-1-phosphonate,
5-[N,N-di(3-hydroxypropyl)-N-hexadecylammonio]-2-hydroxy-pentane-1-sulfat-
e, and combinations thereof.
[0086] Suitable non-ionic surfactants include, for example, mono-
and di-alkanolamides such as, for example, cocamide MEA and
cocamide DEA, amine oxides, alkyl polyglucosides, ethoxylated
silicones, ethoxylated alcohols, ethoxylated carboxylic acids,
ethoxylated amines, ethoxylated amides, ethoxylated alkylolamides,
ethoxylated alkylphenols, ethoxylated glyceryl esters, ethoxylated
sorbitan esters, ethoxylated phosphate esters, glycol stearate,
glyceryl stearate, and combinations thereof. It will be recognized
by one skilled in the art that many of the nonionic surfactants
described herein may act to improve the foaming properties of the
cleansing composition of the multi-product care system, and may
provide a more compact, reduced bubble size or creamy foam.
[0087] The cleansing composition may also include a thickener,
which acts to thicken or increase the viscosity of the cleansing
formulation. A variety of thickeners may be used in the cleansing
compositions described herein. In one embodiment, the thickener may
be a cellulosic thickener or gum. Examples of suitable cellulosic
or gum thickeners include xanthan gum, agar, alginates,
carrageenan, furcellaran, guar, cationic guar, gum arabic, gum
tragacanth, karaya gum, locust bean gum, dextran, starch, modified
starches, gellan gum, carboxymethylcellulose,
hydroxypropylcellulose, hydroyethylcellulose, propylene glycol
alginate, hydroxypropyl guar, amylopectin, cellulose gum, chitosan,
modified chitosan, hydroxypropyl methylcellulose, microcrystalline
cellulose, silica, fumed silica, colloidal silica, dehydroxanthan
gum, non-acrylic based carbomers, and combinations thereof
[0088] Alternately or in addition, the thickener may be an acrylic
based polymer. Non-limiting examples of suitable acrylic based
polymer thickeners include acrylates/C10-C30 alkyl acrylate
crosspolymers, certain carbomers, acrylates copolymers,
aminoacrylates copolymers, and combinations thereof. Examples of
commercially available acrylic based polymer thickeners include
Structure.RTM. Plus (Akzo Nobel, Pasadena, Calif.), which is an
acrylates/aminoacrylates/C10-30 alkyl PEG-20 itaconate copolymer,
Carbopol.RTM. Aqua SF-1 Polymer (Lubrizol Advanced Materials,
Cleveland, Ohio), which is an acrylates copolymer, PEMULENO TR-1
and TR-2 and Carbopol.RTM. ETD 2020 (available from Lubrizol
Advanced Materials), which are acrylates/C10-30 alkyl acrylates
crosspolymers, and the Carbopol.RTM. Ultrez series of polymers
(available from Lubrizol Advanced Materials), which are
carbomers.
[0089] Additional suitable agents for use in the cleansing
composition may include humectants, preservatives, fragrances,
chelating agents, and combinations thereof
[0090] A skin care regime may further include a toning composition.
Toning compositions provide such benefits as closing pores of a
user's skin, restoring the natural pH of the skin (typically, a pH
of from about 5.0 to about 5.5), removing skin impurities (e.g.,
dirt, oils, sebum, make-up, pollutants, and the like), hydrating
the skin, and generally preparing the skin for treatment using a
treatment composition, such as the anti-aging composition of the
present disclosure, and/or an additional treatment composition as
described below.
[0091] Generally, toning compositions for skin care include
astringents, humectants, carriers, and combinations thereof.
Suitable astringents include, for example, ethanol, witch hazel,
rose water, alum, oatmeal, yarrow, bayberry, cold water, rubbing
alcohol, astringent preparations such as silver nitrate, zinc
oxide, zinc sulfate, Burow's solution, tincture of benzoin, and
vegetable substances such as tannic and gallic acid, and
combinations thereof. As used herein, the term "cold water" refers
to water having a temperature below room temperature (approximately
25.degree. C. (77.degree. F.)).
[0092] Suitable humectants include, for example, glycerin, glycerin
derivatives, sodium hyaluronate, betaine, amino acids,
glycosaminoglycans, honey, sorbitol, glycols, polyols, sugars,
hydrogenated starch hydrolysates, salts of PCA, lactic acid,
lactates, and urea. A particularly preferred humectant is
glycerin.
[0093] Carriers for the toning compositions can be any carrier
material typically known in the cosmetic and medical arts as a
basis for ointments, lotions, creams, salves, aerosols, gels,
suspensions, sprays, foams, and the like, and may be used in their
art-established levels. In one particular embodiment, the carrier
is an aqueous carrier. In another embodiment, the carrier is an
alcohol carrier. The alcohol carrier can be any suitable alcohol.
One particularly preferred alcohol is ethanol.
[0094] Other suitable carriers can also be used in the toning
compositions. In certain embodiments, the carriers themselves can
provide the skin care benefit. Non-limiting examples of suitable
carriers include emollients, sterols or sterol derivatives, natural
and synthetic fats or oils, polyols, surfactants, esters,
silicones, and other pharmaceutically acceptable carrier
materials.
[0095] In some embodiments, the skin care regimen includes a
treatment composition for treating the skin in addition to benefits
provided by the composition of the present disclosure. Exemplary
actives for the additional treatment composition may include
actives that are known to have a treating effect on the skin such
as improving the evenness of skin tone and reduction of acne. More
specifically, the treatment agent can be selected from the group
consisting of appearance modifying agents (e.g., exfoliating
agents, skin-firming agents, anti-callous agents, anti-acne agents,
wound care agents, enzyme agents, scar repair agents, humectant
agents), therapeutic agents, pharmaceuticals (e.g., drugs,
anti-oxidants, transdermal drug delivery agents, botanical
extracts, vitamins, magnets, magnetic metals, and foods),
xenobiotics, skin coloration agents (e.g., tanning agents,
lightening agents, and brightening agents, shine control agents and
drugs), shine control agents, colorant agents, surface conditioning
agents (e.g., pH adjusting agents, moisturizers, skin conditioners,
exfoliation agents, shaving lubricants, anti-callous agents,
anti-acne agents, anti-aging agents, wound care agents, skin
lipids, enzymes, scar care agents, humectants, powders, botanical
extracts, and drugs) external analgesic agents, anti-inflammatory
(e.g., anti-irritant agents, anti-allergy agents, wound care
agents, transdermal drug delivery, and drugs), fragrances, odor
neutralizing agents, soothing agents, calming agents, botanical
extracts (e.g., peppermint oil, eucalyptol, eucalyptus oil,
camphor, and tea tree oil), peptides, natural and synthetic fats or
oils, moisture absorbers, and combinations thereof.
[0096] In one embodiment, in addition to the compositions described
above, the skin care regimen includes a finishing composition. As
with the other compositions, it should be understood that the
various active ingredients described herein can be used in any of
the products of the multi-product care regimen without departing
from the scope of the disclosure.
[0097] When included, a finishing composition comprises a finishing
agent that typically delivers moisturization, skin protection, or a
moisture-barrier for sealing in moisture to the user. Specifically,
the finishing agent can be any moisturizing agent, skin protectant,
and/or moisture-barrier enhancing agent known in the art.
[0098] Additionally, the finishing agent may be capable of
providing aesthetic benefits such as skin smoothing or a powdery
feel. Examples of additional suitable finishing agents include skin
conditioning agents (e.g., pH adjusting agents, moisturizers, skin
conditioners, exfoliation agents, shaving lubricants, skin-firming
agents, anti-callous agents, anti-acne agents, anti-aging agents,
anti-wrinkle agents, anti-dandruff agents, wound care agents, skin
lipids, enzymes, scar care agents, humectants, powders, botanical
extracts, and drugs), fragrances, botanical extracts, powders, and
combinations thereof
[0099] It should be understood that the various active ingredients
can be used in any of the products of the multi-product care
regimen without departing from the scope of the disclosure. The
specific active ingredients of the various products will depend
upon the end daily regimen desired.
[0100] It should be understood by a skilled artisan that, while
skin care systems will be discussed herein, regimes using the
compositions of the present disclosure can be used for various
other daily regimens comprising steps to cleanse, treat, moisturize
and protect the skin. It is understood that skin care regimens can
combine all of these steps, some of these steps, or have multiple
iterations of the same steps so as to provide desired benefits to
the skin.
[0101] This written description uses examples to disclose the
invention, including the best mode, and also to enable any person
skilled in the art to practice the invention, including making and
using the compositions and performing any incorporated methods. The
patentable scope of the present disclosure is defined by the
claims, and may include other examples that occur to those skilled
in the art. Such other examples are intended to be within the scope
of the claims if they have structural elements that do not differ
from the literal language of the claims, or if they include
equivalent structural elements with insubstantial differences from
the literal languages of the claims.
[0102] The disclosure will be more fully understood upon
consideration of the following non-limiting Examples.
EXAMPLE 1
Cell Viability
[0103] In this Example, a cell viability test was conducted to test
cytotoxicity.
[0104] Primary human dermal fibroblasts were grown to confluence in
growth media (DMEM+10% FBS) at 37.degree. C. in 5% CO.sub.2
atmosphere. Samples of cells (n=3) were then treated with
increasing concentrations of UNDARINE.TM. or ECOBIDENS.TM. in
growth media (DMEM (Life Technologies, Grand Island, N.Y.) +2%
Fetal Bovine Serum (FBS) (Life
Technologies)+1.times.antibiotic/antimycotic (AA)) for either 1 or
24 hours. After the treatment, cells were washed twice in PBS, let
recover in growth media (DMEM+10% FBS) for 12 hours at 37.degree.
C. in 5% CO.sub.2 atmosphere, and then incubated with
ALAMARBLUE.RTM. (Life Technologies, Grand Island, N.Y.) for 2 hours
at 37.degree. C. in 5% CO.sub.2 atmosphere. The ALAMARBLUE.RTM.
assay incorporates a fluorometric/colorimetric growth indicator
based on detection of metabolic activity. Specifically, the system
incorporates an oxidation-reduction (REDOX) indicator that both
fluoresces and changes color in response to chemical reduction of
growth medium resulting from cell growth.
[0105] The mitochondrial uncoupling drug, Carbonyl cyanide
3-chlorophenylhydrazone (CCCP) (commercially available from
Sigma-Aldrich, St. Louis, Mo.), was utilized as a positive control
for cytotoxicity. Further, non-treated samples of cells were
utilized as a negative control.
[0106] The intensities of fluorescent emission were recorded using
SpectraMax.degree. M5 (Molecular Devices, Sunnyvale, California) at
585 nm. Viability was determined as follows:
Viability (%)=((fluorescence intensity at 585 nm of the treated
sample)/(fluorescence intensity at 585 nm of the non-treated
sample)).times.100.
[0107] As demonstrated in FIG. 1, neither of the tested active
agents significantly reduced cellular viability at concentrations
of 1% or 3%.
EXAMPLE 2
[0108] In this Example, an Undaria extract was analyzed for its
ability to protect fibroblasts against oxidative stress.
[0109] Primary fibroblasts from an abdominal explant of a 51-year
old woman were grown to confluence in growth media (DMEM+10%
FBS+1.times. antibiotic/antimycotic (AA)) and then treated for 20
hours with 3% v/v UNDARINE.TM. in growth media (DMEM (Life
Technologies, Grand Island, N.Y.)+2% Fetal Bovine Serum (FBS) (Life
Technologies)+1.times.antibiotic/antimycotic (AA)). Cells were
washed and then a sample of cells (n=4) was further treated with
200 .mu.M H.sub.2O.sub.2 for 1 hour at 37.degree. C. A second
sample of cells (n=4) was not treated with UNDARINE.TM., but was
treated with H.sub.2O.sub.2. After removing H.sub.2O.sub.2, cells
were allowed to recover in media at 37.degree. C. in 5% CO.sub.2
atmosphere. A sample of untreated cells (n=4) was used as a
control. After 12 hours, cells were incubated in PBS+10%
ALAMARBLUE.RTM. (Life Technologies, Grand Island, N.Y.) for 1 hour
at 37.degree. C. in 5% CO.sub.2 and fluorescence emission was
measured using SpectraMax.RTM. M5 (Molecular Devices, Sunnyvale,
Calif.) (560EX nm/590 nm). The results are shown in FIG. 2.
[0110] As shown in FIG. 2, UNDARINE.TM. protected the cells from
H.sub.2O.sub.2 exposure.
EXAMPLE 3
[0111] In this Example, an Undaria extract was analyzed for its
ability to protect fibroblasts against H.sub.2O.sub.2-induced
apoptosis.
[0112] Human dermal fibroblasts were grown to confluence in growth
media (DMEM+10% FBS) and then treated for 12 hours with 3% v/v
UNDARINE.TM. in growth media (DMEM (Life Technologies, Grand
Island, N.Y.)+2% Fetal Bovine Serum (FBS) (Life
Technologies)+1.times.antibiotic/antimycotic (AA)). Cells were
washed twice with PBS and then a sample of cells (n=4) was further
treated with 200 .mu.M H.sub.2O.sub.2 for 1 hour at 37.degree. C. A
second sample of cells (n=4) was not treated with UNDARINE.TM., but
was treated with H.sub.2O.sub.2. A sample of untreated cells (n=4)
was used as a control. After further washing with PBS, cells were
allowed to recover in media at 37.degree. C. in 5% CO.sub.2
atmosphere. After 12 hours, cells were stained with the apoptotic
marker Annexin V Alexa Fluor 488 (Life Technologies, Grand Island,
N.Y.) (fluorescent emission reported as Annexin). Annexin V is used
as an industry standard as a marker of early apoptosis.
Particularly, compositions and/or compounds that reduce its
expression would be considered highly favorable/effective at
improving skin function and appearance. The results are shown in
FIGS. 3A-3D.
[0113] As shown in FIGS. 3A-3D, UNDARINE.TM. prevented the increase
in an early apoptotic marker caused by the H.sub.2O.sub.2 insults.
Also, FSC and HSC values (size and granularity) indicated a
decrease in percentage of apoptotic cells in UNDARINET.TM.-treated
samples.
EXAMPLE 4
[0114] In this Example, the effect of a Bidens extract on
H.sub.2O.sub.2 or UV-induced photodamage in human dermal
fibroblasts was analyzed.
[0115] Primary fibroblasts were isolated from an abdominal skin
explant of a 32-year old Caucasian female. The fibroblasts were
cultured at 37.degree. C. in 5% CO.sub.2 atmosphere in 24-well
plates including growth media (DMEM (Life Technologies, Grand
Island, N.Y.)+2% Fetal Bovine Serum (FBS) (Life
Technologies)+1.times.antibiotic/antimycotic (AA)) up to 100%
confluence.
[0116] After culturing to 100% confluence, the cells were treated
for 20 hours with 3% v/v ECOBIDENS.TM. in growth media. The cells
were then washed twice with PBS, and either treated with 200 .mu.M
H.sub.2O.sub.2 (n=4) in growth media for one hour at 37.degree. C.
in 5% CO.sub.2 atmosphere or submerged in 0.5 ml of PBS and exposed
to 100 mJ UV/cm.sup.2 (n=4).
[0117] After washing twice with PBS, the cells were then left to
recover for approximately 12 hours in media at 37.degree. C. in 5%
CO.sub.2 atmosphere. The media was then collected and stored at
-80.degree. C. for further cytokine analysis and it was replaced
with PBS+10% ALAMARBLUE.RTM. (Life Technologies). After incubating
for one hour at 37.degree. C. in 5% CO.sub.2 atmosphere, the
fluorescence emission was measured using SpectraMax.RTM. M5
(Molecular Devices, Sunnyvale, Calif.) (560EX nm/590 nm). The
results are shown in in FIGS. 4 and 5.
[0118] Using the fluorescent emission, change in the viability of
the cells may be calculated by comparing the ratio of the
fluorescent emission of the treated cells to the untreated control
cells. As shown by the results, cells treated with ECOBIDENS.TM.
displayed a notably higher viability than non-pretreated control
cells.
EXAMPLE 5
[0119] In this Example, a Bidens extract was analyzed for its
ability to protect fibroblasts against UV- or
H.sub.2O.sub.2-induced apoptosis.
[0120] Human dermal fibroblasts were grown to confluence in growth
media (DMEM+10% FBS) and then treated for 12 hours with 3% v/v
ECOBIDENS.TM. in growth media (DMEM (Life Technologies, Grand
Island, N.Y.)+2% Fetal Bovine Serum (FBS) (Life
Technologies)+1.times.antibiotic/antimycotic (AA)). Cells were
washed with PBS and then samples of cells were further treated with
either 200 .mu.M H.sub.2O.sub.2 for 1 hour at 37.degree. C. or
irradiated with 100 mJ UV/cm.sup.2. A sample of untreated cells was
used as a control. Another sample of cells was not treated with
ECOBIDENS.TM., but was treated with either H.sub.2O.sub.2 or UV
(n=4). After further washing with PBS, cells were allowed to
recover in media at 37.degree. C. in 5% CO.sub.2 atmosphere. After
12 hours, cells were stained with the apoptotic marker Annexin V
Alexa Fluor 488 (Life Technologies, Grand Island, N.Y.)
(fluorescent emission reported as Annexin). The results are shown
in FIGS. 6A-6F. Results represent two independent experiments
analyzing 10,000 cells/reading.
[0121] As shown in FIGS. 6A-6F, ECOBIDENS.TM. prevented the
increase in early apoptotic markers caused by the UV and/or
H.sub.2O.sub.2 insults. Also, FSC and HSC values (size and
granularity) indicated a decrease in percentage of apoptotic cells
in ECOBIDENS.TM.-treated samples.
EXAMPLE 6
[0122] In this Example, a Bidens extract was analyzed for its
ability to minimize the decrease in cellular glutathione (GSH)
concentration due to UV irradiation or H.sub.2O.sub.2-induced
stress.
[0123] Human dermal fibroblasts were grown to confluence in growth
media (DMEM+10% FBS) and then treated for 24 hours with 3% v/v
ECOBIDENS.TM. in growth media (DMEM (Life Technologies, Grand
Island, N.Y.)+2% Fetal Bovine Serum (FBS) (Life
Technologies)+1.times.antibiotic/antimycotic (AA)). Cells were
washed with PBS and then samples of cells were further treated with
either 200 .mu.M H.sub.2O.sub.2 (n=4) for 1 hour at 37.degree. C.
or irradiated with 100 mJ UV/cm.sup.2 (n=4). A sample of untreated
cells (n=4) was used as a control. Another sample of cells was not
treated with ECOBIDENS.TM., but was treated with either
H.sub.2O.sub.2 or UV (n=4/treatment). After further washing the
cells with PBS three times, cells were allowed to recover for
approximately 20 hours in media (DMEM+10% FBS) at 37.degree. C. in
5% CO.sub.2 atmosphere.
[0124] Cells were harvested by trypsinization and centrifuged in
1.5 ml tubes at 1500 rpm for 5 minutes at 4.degree. C. to pellet
the cells. The supernatant was discarded and cells were resuspended
in PBS. Cells were then snap frozen by immersion in liquid nitrogen
and thawed quickly in a 37.degree. C. water bath. Four cycles of
freeze-thaw were undertaken. The tubes were centrifuged at
10,000.times.g at 4.degree. C. for 15 minutes. The supernatant was
transferred to new 1.5 ml tubes and stored at -80.degree. C. for
reduced glutathione (GSH) analysis using the SENSOLYTE.RTM.
Glutathione Cellular Assay Kit (AnaSpec, Inc., Fremont, Calif.).
The results of the GSH analysis are shown in FIG. 7.
[0125] As shown in FIG. 7, both the UV and H.sub.2O.sub.2 insults
caused a decrease in cellular GSH levels. The impact of
H.sub.2O.sub.2 and UV insults on GSH levels, however, was minimized
in the sample cells treated with ECOBIDENS.TM..
EXAMPLE 7
[0126] In this Example, the activity of a Bidens extract on
H.sub.2O.sub.2 concentration was analyzed.
[0127] Primary human dermal fibroblasts were grown to confluence in
growth media (DMEM+10% FBS) and then treated for 12 hours with 3%
v/v ECOBIDENS.TM. in growth media (DMEM (Life Technologies, Grand
Island, N.Y.)+2% Fetal Bovine Serum (FBS) (Life
Technologies)+1.times.antibiotic/antimycotic (AA)). Three control
samples were left untreated. N=4 for each treated and untreated
sample. Cells of all samples were then washed twice with PBS. One
control sample was treated at 37.degree. C. with 200 .mu.M
H.sub.2O.sub.2; one control sample was treated at 37.degree. C.
with 200 .mu.M H.sub.2O.sub.2 and catalase (available as Hydrogen
Peroxide Cell-Based Assay Kit as used according to the
manufacturer's instructions (Cayman Chemicals Company, Ann Arbo,
Mich.)). An ECOBIDENS.TM.-treated sample was also treated at
37.degree. C. with 200 .mu.M H.sub.2O.sub.2. After 1 hour of
treatment with H.sub.2O.sub.2, H.sub.2O.sub.2 concentration in the
sample was measured with a Hydrogen Peroxide Cell-Based Assay Kit
(Cayman Chemicals Company, Ann Arbo, Mich.). More particularly, the
kit measured fluorescence emission as being proportional to
H.sub.2O.sub.2 concentration.
[0128] As shown in FIG. 8, pretreatment with a Bidens extract does
not reduce the concentration of H.sub.2O.sub.2 in a sample. That
is, as the Bidens extract is washed prior to treatment with
H.sub.2O.sub.2, it has now been shown that residual amounts of the
extract are not causing the reduction in H.sub.2O.sub.2. By
contrast, it is believed that the extract enhances the cells own
defense mechanisms to reduce the concentration of
H.sub.2O.sub.2.
EXAMPLE 8
[0129] In this Example, extracts were analyzed for their ability to
enhance cell resilience to UV irradiation and/or oxidative
stress.
[0130] Primary human keratinocytes were grown to confluence in
growth media (EpiLife.RTM. Medium, Life Technologies, Grand Island,
N.Y., with 60 .mu.M calcium (Gibco.RTM., Life Technologies, Grand
Island, N.Y.), supplemented with EpiLife.RTM. Defined Growth
Supplement (EDGS) (Life Technologies, Grand Island, N.Y.) and
1.times.Antibiotics/antimycotics) and then treated for 12 hours
with 3%, 6%, or 9% v/v of either UNDARINE.TM. or ECOBIDENS.TM.. N=4
for each treated and untreated sample. One control sample was left
untreated and another control sample was treated with 5 .mu.M of
rotenone. Cells were washed and then samples of cells were further
treated with 200 .mu.M H.sub.2O.sub.2 for 1 hour at 37.degree. C.
or irradiated with 100 mJ/cm.sup.2 UV. Cells were allowed to
recover in media (DMEM+10% FBS) at 37.degree. C. in 5% CO.sub.2
atmosphere. After 16 hours, cells were incubated in PBS+10%
ALAMARBLUE.RTM. (Life Technologies, Grand Island, N.Y.) for 1 hour
at 37.degree. C. in 5% CO.sub.2 and fluorescence emission was
measured using SpectraMax.RTM. M5 (Molecular Devices, Sunnyvale,
Calif.) (560EX nm/590 nm). The results are shown in FIGS. 9 and
10.
[0131] As shown in FIGS. 9 and 10, both UNDARINE.TM. and
ECOBIDENS.TM. protected the cells from H.sub.2O.sub.2 and/or UV
exposure. It is believed that both UNDARINE.TM. and ECOBIDENS.TM.
enhanced keratinocyte resilience to oxidative stress caused by
H.sub.2O.sub.2 treatment and/or UV irradiation by enhancing
cellular mitochondrial activity and/or antioxidant cellular
response.
[0132] The above Examples show that both Undaria and Bidens
extracts are effective in protecting cells from photo-damage and
oxidative stress. Accordingly, compositions including these
extracts can be topically applied to protect cells from UV exposure
and excessive production of ROS, thereby resulting in a more
youthful appearance of aged skin of the face and body. It is
believed that these protective effects are due to Undaria and
Bidens being capable of enhancing cellular defense mechanisms, and
in particular, cellular mitochondrial activity and antioxidant
cellular response, thereby minimizing cell degeneration and death.
These effects have proven to slow and/or reduce the process of skin
aging and its the symptoms (e.g., skin thinning, sagging,
wrinkling, fragility, and loss of skin resiliency).
* * * * *