U.S. patent application number 11/100905 was filed with the patent office on 2005-11-03 for topical anti-cancer compositions and methods of use thereof.
Invention is credited to Conney, Allan H., Huang, Mou-Tuan, Paine, Christine, Seiberg, Miri, Shapiro, Stanley S..
Application Number | 20050244523 11/100905 |
Document ID | / |
Family ID | 27804381 |
Filed Date | 2005-11-03 |
United States Patent
Application |
20050244523 |
Kind Code |
A1 |
Seiberg, Miri ; et
al. |
November 3, 2005 |
Topical anti-cancer compositions and methods of use thereof
Abstract
Described are skin-care compositions containing non denatured
soy products and optionally other anti-cancer or anti-aging agents.
The compositions can be applied topically to reduce the risk of
UV-induced cutaneous tumors.
Inventors: |
Seiberg, Miri; (Princeton,
NJ) ; Shapiro, Stanley S.; (Roseland, NJ) ;
Paine, Christine; (Hoboken, NJ) ; Conney, Allan
H.; (Princeton, NJ) ; Huang, Mou-Tuan;
(Englewood Cliffs, NJ) |
Correspondence
Address: |
DANN, DORFMAN, HERRELL & SKILLMAN
1601 MARKET STREET
SUITE 2400
PHILADELPHIA
PA
19103-2307
US
|
Family ID: |
27804381 |
Appl. No.: |
11/100905 |
Filed: |
April 7, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11100905 |
Apr 7, 2005 |
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10108248 |
Mar 27, 2002 |
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10108248 |
Mar 27, 2002 |
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09698454 |
Oct 27, 2000 |
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Current U.S.
Class: |
424/757 |
Current CPC
Class: |
A61Q 17/04 20130101;
A61K 36/48 20130101; A61K 2800/782 20130101; A61K 8/44 20130101;
A61K 8/494 20130101; A61K 8/671 20130101; A61K 2800/51 20130101;
A61Q 19/00 20130101; A61K 8/347 20130101; A61Q 19/02 20130101; A61K
45/06 20130101; A61Q 19/08 20130101; A61K 8/9789 20170801; Y10S
514/887 20130101; A61K 2800/522 20130101; A61P 35/00 20180101; A61K
36/48 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/757 |
International
Class: |
A61K 035/78 |
Claims
1-49. (canceled)
50. A method for preventing and reducing the risk of skin cancer
comprising topical application of at least one composition
comprising a non-denatured, soy product in an amount of from about
0.01-99% by weight in a carrier.
51. The method of claim 50, wherein said composition is applied at
least once daily on a continuous basis.
52. The method of claim 50, wherein said composition is applied at
least twice daily for at least eight weeks and at least once daily
on a continuous basis thereafter.
53. The method of claim 50, wherein a first composition comprising
non-denatured, soy product in an amount of 0.05-80% by weight is
applied for at least about four to about ten weeks followed by
topical application of a second composition comprising
non-denatured, soy product in an amount of 0.01-20% by weight on a
daily basis thereafter.
54. The method of claim 50, wherein said composition further
comprises a cosmetically acceptable vehicle.
55. The method of claim 50, wherein said composition further
comprises at least one compound selected from the group consisting
of anti-oxidants, sunscreens, moisturizers, bleaching agents,
depigmentation agents, darkening agents, surfactants, foaming
agents, conditioners, humectants, fragrances, anti-aging agents,
anti-inflammatory agents, and anti-cancer agents.
56. The method of claim 55, wherein said composition further
comprises at least one anti-inflammatory agent.
57. The method of claim 55, wherein said composition further
comprises at least one anti-cancer agent.
58. The method of claim 55, wherein said composition further
comprises at least one anti-oxidant.
59. The method of claim 55, wherein said composition further
comprises at least one sunscreen.
60. The method of claim 50, wherein said non-denatured, soy product
is selected from the group consisting of soymilk, soybean powder,
soymilk powder, and Kunitz-type soybean trypsin inhibitor.
61. The method of claim 60, wherein said non-denatured, soy product
is soymilk.
62. The method of claim 60, wherein said non-denatured, soy product
is soybean powder or soymilk powder.
63. The method of claim 60, wherein said non-denatured, soy product
is Kunitz-type soybean trypsin inhibitor.
64. The method of claim 61, wherein said soymilk is in an amount of
0.05% to 80%, said composition further comprising from about 0.1%
to about 20% emulsifier and a preservative in an effective
amount.
65. The method of claim 64, wherein said composition further
comprises at least one compound selected from the group consisting
of anti-oxidants, sunscreens, moisturizers, bleaching agents,
depigmentation agents, darkening agents, surfactants, foaming
agents, conditioners, humectants, fragrances, anti-aging agents,
anti-inflammatory agents, and anti-cancer agents.
66. The method of claim 65, wherein said composition further
comprises at least one anti-oxidant.
67. The method of claim 65, wherein said compositions further
comprises at least one anti-cancer agent.
68. The method of claim 65, wherein said composition further
comprises at least one sunscreen.
69. The method of claim 65, wherein said composition further
comprises at least one anti-inflammatory agent.
70. The method of claim 62, wherein said soybean powder or soymilk
powder is in an amount of 0.05% to 80%, said composition further
comprising from about 0.1% to about 20% emulsifier and a
preservative in an effective amount.
71. The method of claim 70, wherein said composition further
comprises at least one compound selected from the group consisting
of anti-oxidants, sunscreens, moisturizers, bleaching agents,
depigmentation agents, darkening agents, surfactants, foaming
agents, conditioners, humectants, fragrances, anti-aging agents,
anti-inflammatory agents, and anti-cancer agents.
72. The method of claim 71, wherein said composition further
comprises at least one anti-oxidant.
73. The method of claim 71, wherein said compositions further
comprises at least one anti-cancer agent.
74. The method of claim 71, wherein said composition further
comprises at least one sunscreen.
75. The method of claim 71, wherein said composition further
comprises at least one anti-inflammatory agent.
76. The method of claim 63, wherein said Kunitz-type soybean
trypsin inhibitor is in an amount of from about 0.01 to about 10%,
said composition further comprising from about 0.1 to about 20%
emulsifier and a preservative in an effective amount.
77. The method of claim 76, wherein said composition further
comprises at least one compound selected from the group consisting
of anti-oxidants, sunscreens, moisturizers, bleaching agents,
depigmentation agents, darkening agents, surfactants, foaming
agents, conditioners, humectants, fragrances, anti-aging agents,
anti-inflammatory agents, and anti-cancer agents.
78. The method of claim 77, wherein said composition further
comprises at least one anti-oxidant.
79. The method of claim 77, wherein said compositions further
comprises at least one anti-cancer agent.
80. The method of claim 77, wherein said composition further
comprises at least one sunscreen.
81. The method of claim 77, wherein said composition further
comprises at least one anti-inflammatory agent.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application is a continuation in part of U.S.
Application Ser. No. 09/698,454 filed Oct. 27, 2000, the entire
disclosure of which is incorporated by reference herein.
FIELD OF THE INVENTION
[0002] This invention relates to compositions containing
non-denatured soy products, or soy trypsin inhibitors, and
optionally additional anti-cancer or cosmetically active agents.
These compositions can be applied topically to reduce the risk of
UV-induced cutaneous tumors.
BACKGROUND OF THE INVENTION
[0003] Skin, the largest organ of the human body, is continuously
exposed to environmental insults such as smoke, pollution, and
ultraviolet (UV) irradiation. The thinning of the ozone layer,
which is expected to progress for at least several decades, reduces
a major barrier to the passage of ultraviolet-B radiation (UVB)
through the atmosphere. UVB, that is., light whose wavelength is in
the range between about 280 and about 320 nm, is the main cause of
sunburn, tanning, aging of the skin, and skin cancer.
[0004] The non-melanoma skin cancers (NMSC), including basal-cell
and squamous-cell carcinoma, are the most common types of cancer
among Caucasian populations. The incidence of NMSC has increased
worldwide over the last few decades. Increased recreational and
occupational sunlight exposure is commonly regarded as one of the
reasons for the higher incidence of cutaneous cancers. The increase
in UVB exposure associated with the thinning of the ozone layer is
another significant factor. Mortality from NMSC is low, but the
estimated recurrence rate of about 50% after five years and the
local invasiveness of this type of cancer result in high medical
costs. Therefore, NMSC constitutes a substantial public health
concern. (Reviewed in Holick and Kligman, editors: Biologic effects
of light. Walter de Gruyter, Berlin and New York, 1992).
[0005] Photo-carcinogenesis results from a complex interplay of
simultaneous and sequential biochemical events. These events,
initiated by irradiation of an organism with UV light of an
appropriate wavelength, include the formation of DNA
photo-products, inaccuracies in DNA repair, mutation of
proto-oncogenes and tumor suppressor genes, and UV-induced
production of radical species which produce subsequent effects on
existing mutations and independently induce further mutations. In
addition, other epigenetic events such as immunological responses,
antioxidant defenses, and dietary factors may influence the course
of carcinogenesis. (Black, H. S., deGruijl, F. R., Forbes P. D.,
Cleaver, J. E., Ananthaswamy, H. N., deFabo, E. C., Ullrich, S. E.,
Tyrrell, R. M., Photo-carcinogenesis: an overview. J. Photochem.
Photobiol. B 40:1, 29-47, August, 1997).
[0006] The skin possesses an elaborate antioxidant defense system
to deal with UV-induced oxidative stress. Excessive exposure to UV
radiation, however, can overwhelm the cutaneous antioxidant
capacity, leading to oxidative damage and ultimately to skin cancer
and premature skin aging. Therefore, one strategy for
photo-protection is to support the endogenous antioxidant system by
induction or transdermal delivery of antioxidant enzymes or
nonenzymatic antioxidants. Antioxidants such as glutathione,
alpha-tocopherol, ascorbate and beta-carotene have been found to be
very effective in photoprotection. Components of the antioxidant
pathway have also been identified and applied to the skin of
patients. Although skin treatments with single components of the
antioxidant system such as vitamin E were successful against a wide
variety of types of photodamage, they were not shown to affect the
progression of UV-induced tumors. The most promising results were
obtained in studies combining several compounds, which often
resulted in synergy between the protective effects. (Steenvoorden
D. D., van Henegouwen G. M., The use of endogenous antioxidants to
improve photoprotection, J. Photochem. Photobiol., B 41:1-2, 1-10,
November, 1997).
[0007] Epidemiological studies suggest that components of
vegetables, particularly legumes, are beneficial in lowering the
incidence rates of many types of cancer. For example, the rates of
breast, colon and prostate cancer are significantly lower among the
inhabitants of most countries of the Pacific Basin, but offspring
of Pacific Basin natives who have migrated to the United States
develop the common Western cancers at approximately the same rate
as native Westerners. Such epidemiological studies suggest that
dietary and other environmental factors, rather than genetic
differences, contribute more significantly to the risk of
susceptibility to these cancers. The high consumption of soybean
products in Pacific Basin countries, such as Japan, implicates diet
as one factor contributing to the relatively extremely low rates of
cancer mortality in these countries. (E.g., Wu et al., Soy intake
and risk of breast cancer in Asians and Asian Americans. Am. J.
Clin. Nutr. 68: 6 Suppl., 1437S-1443S, December, 1998).
[0008] Soybeans are a rich source of isoflavones, which possess
weak estrogenic activity. Genistein, the main soybean isoflavone,
is a specific inhibitor of protein tyrosine kinases and of other
enzymes involved in signal transduction. Genistein has been shown
to suppress the growth of numerous cancer cells in vitro, and to
protect animals in experimental carcinogenesis models from
developing both hormone- and non-hormone related cancers. (Reviewed
in A. R. Kennedy, Chemopreventive agents: Protease inhibitors,
Pharmacology Theories 78 (3), 167-209), 1998 and in Messina et al.,
Soy intake and cancer risks: A review of the in vitro and in vivo
data, Nutrition and Cancer 21 (2), 113-131, 1994).
[0009] Soybeans also contain a number of protease inhibitors such
as BBI and STI. It is important to note that soy foods do not
contain high concentrations of active STI and BBI, because these
protease inhibitors block the action of trypsin and other enzymes
needed for protein digestion. Although STI is denatured by cooking,
heat alone does not inactivate BBI, and consumption of soy products
containing high levels of these protease inhibitors leads to
serious digestive problems, chronic deficiency in amino acid
uptake, and cancer. Indeed, the Chinese did not serve soybeans as
food until fermentation techniques were developed to destroy the
anti-digestive properties of the soy foods (2nd century B.C.E.).
During the production of soy foods today, pureed soybeans are
soaked in an alkaline solution and then pressure-heated to
115.sup.EC in order to denature the protease inhibitors as much as
possible.
[0010] Limtrakul et al. attempted to identify a safe level of soy
proteins for nutritional consumption, which would be beneficial in
the prevention of cancer. Skin tumors were chemically induced in
mice, which were fed soy protein isolate (SPI) exclusively, and in
mice which were fed SPI supplemented with soymilk proteins (SMP).
It was reported that "the percentage of tumor-bearing mice and the
volume of tumor tended to be lower in the mice on the SMP diet".
Life Sciences 1993, 53, 1591-1596. When defatted soybeans are
treated first with alkaline, then with acid solution, SPI is the
precipitate and SMP is the supernatant. The Limtrakul study shows
the potential of soy proteins to affect skin cancer progression,
when the proteins are orally consumed. However, it was also
emphasized that higher levels of dietary intake of SMP would result
in major health problems.
[0011] It is clear that a need exists for safe, efficacious and
economical agents that prevent or reduce incidence of cancer,
particularly for NMSC, which may be simply and conveniently
administered. Further, economical and prophylactic compositions and
methods for the reduction, prevention or inhibition of the
progression of UV-induced cutaneous tumors are highly desirable.
Since topical application is very simple and convenient,
incorporating compositions that reduce skin cancer incidence into a
skin-care product would be extremely advantageous. While sunscreens
are known to reduce the damage resulting from UV exposure during
the period of their application, there is a need for a skin care
product that could also slow the progression of already-initiated
photocarcinogenic processes. It is an object of the invention to
provide such a product.
SUMMARY OF THE INVENTION
[0012] The present invention provides a method of reducing the risk
of developing UV-induced tumors of the skin of a mammal by
topically applying a skin-care composition, preferably to an
individual who has already been exposed to or irradiated with UV
light. A method of reducing the growth rate of UV-induced cutaneous
tumors by topically applying the skin-care composition is also
provided, as is a method of preventing the progression of cancer by
the same means.
[0013] The skin care composition for use in the methods of the
invention is formulated for the topical delivery of a non-denatured
soy product (e.g., to a mammal such as a human) and comprises a soy
product (e.g., a non-denatured soymilk or soybean powder or soybean
trypsin inhibitor) and a vehicle. The composition may optionally
comprise other anti-cancer or cosmetically active agents. Certain
skin care compositions appropriate for use in the present invention
have been described in U.S. patent application Ser. Nos.09/110,409,
09/621,565 and 09/698,454, filed Jul. 6, 1998, Jul. 21, 2000 and
Oct. 27, 2000, respectively, and in International Application No.
WO99/04752. Each of the foregoing patent documents is incorporated
herein by reference.
[0014] Other features and advantages of the present invention will
be apparent to those of skill in the art in light of the following
description and claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0015] FIGS. 1A-1C are a series of graphs showing the inhibitory
effects of STI and BBI on the development of skin tumors in SKH-1
mice previously treated with ultraviolet B light. (BSA--bovine
serum albumin; BBI--Bowman-Birk Inhibitor; STI--Soy Bean Trypsin
Inhibitor)
[0016] FIGS. 2A-2C are a series of graphs showing the inhibitory
effect of Soymilk on the development of skin tumors in SKH-1 mice
previously treated with ultraviolet B light. The inhibitory effects
of heat-denatured soymilk vs. non-denatured soymilk are
compared.
[0017] FIGS. 3A-3C are a series of graphs showing the inhibitory
effects caffeine on the development of skin tumors in SKH-1 mice
previously treated with ultraviolet B light.
[0018] FIGS. 4A-4C are three photographs showing the reduction in
size and number of UV-induced tumors among mice treated topically
with non-denatured soymilk (FIG. 4A) compared to mice treated with
heat-denatured soymilk (FIG. 4B) or water (FIG. 4C).
[0019] FIGS. 5A and 5B are a pair of photographs showing the
reduction in size and number of UV-induced tumors among mice
treated topically with a solution of caffeine in acetone (FIG. 5B)
compared to mice treated with acetone alone (FIG. 5A).
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0020] It is believed that one skilled in the art can, based upon
the description herein, utilize the present invention to its
fullest extent. The following specific embodiments are to be
construed as merely illustrative and not limitative of the
remainder of the disclosure. All publications, patent applications,
patents, and other references mentioned herein are incorporated by
reference in their entirety.
[0021] The present invention is directed to soy-containing
compositions and methods of use thereof in the prevention and
reduction of the risk of skin cancer. The novel compositions of
this invention contain legume products, and preferably soy
products, that may be in the form of a fluid (e.g., soymilk) or a
solid (e.g., a soybean powder or soymilk powder). What is meant by
"soy product" is a substance derived from the soybean, containing
the ingredients naturally found in soybeans, at the relative
concentrations as found in the beans, excluding water content. In
one embodiment, the soy product is a non denatured soy product.
[0022] "Denaturation" is defined in the Bantam Medical Dictionary
(1990 edition) as "the change in the physical and the physiological
properties of a protein, that are brought about by heat, X-rays or
chemicals. These changes include loss of activity (in the case of
enzymes) and loss (or alteration) of antigenicity (in the case of
antigens)".
[0023] What is meant by "non-denatured soy product" is a soy
product in which the processing for the derivation of such soy
product (e.g., the temperature, extraction media) did not eliminate
its protease inhibitory activity. In one embodiment, the
non-denatured state of the soy product of this invention is
measured by the presence of an intact soybean trypsin inhibitor
(STI) protein.
[0024] In another embodiment, the soy product is soymilk. One way
to make soymilk is to soak the soybeans in deionized or purified
water for several hours, and grind them after they were fully
hydrated, with the addition of small quantities of water. (The
grinding process allows the soybean milk to be extracted). After
collection, the soybean milk may be filtered to remove any residual
parts of the bean husk. The soymilk used in this invention can be
fresh soymilk as described above, or may be made from soybean
powder and water. The soybean powder is milled from soybeans and
may also be lyophilized, spray dried, or freeze-dried and the
resulting soymilk may or may not be filtered. Soymilk prepared by
these methods may have from about 1 to about 90% by weight dry
soybean powder. Another example is the use of soymilk powder, made
from lyophilized, spray dried or freeze-dried soymilk, with the
addition of water and finished with or without filtration or
homogenization.
[0025] Other methods of soybean extraction could also be used to
create the active ingredients used in this invention. In one
example, the active ingredients could be extracted from ground
soybeans using ethanol/water mixtures, followed by the removal of
the ethanol from the extract, in such ways that the protease
inhibitory activity of the soybean will be retained.
[0026] The compositions of the present invention may contain from
about 1% to about 99%, by weight, of the soy product. For example,
when a liquid soy product (e.g., soymilk) is used, the composition
may contain from about 50% to about 99%, by weight, (e.g., from
about 70% to about 99%) of the liquid soy product. For example,
when a solid soy product (e.g., soybean powder or soymilk powder)
is used, the composition may contain from about 1% to about 50%, by
weight (e.g., from about 2% to about 30%, by weight) of the solid
soy product. Compositions comprising solid soy products may also
comprise water (e.g., distilled water or water contained within
soymilk) to form a liquid base for the composition (e.g., to form a
cream, lotion, injectable solution or gel). Such compositions may
comprise from about 50% to about 98%, by weight (e.g., from about
70% to about 98%, by weight) of water. While not limited to these
methods of administration, the compositions of this invention may
be delivered topically, orally, or parenterally, although topical
administration is preferred.
[0027] The soy products useful in this invention may be produced
from all soybean species, regardless of their geographic origin,
sun exposure, harvest time and the like. However, specific strains,
geographic origins or growth conditions might be preferred. These
include soybean strains or other legume strains particularly rich
in their trypsin inhibitor (e.g. STI, LTI, BBI) content or strains
in which, under the proper growth conditions trypsin inhibitor
enrichment occurs in the bean. It should be noted that the legume
products useful in the compositions of this invention have a
distinctive odor, which may be tolerable in some cultures, but is
undesired in others. If necessary, the odor of the compositions of
this invention can be reduced by using soybean products derived
from specific strains of soybeans known to be less odiferous,
including, but not limited to, lipoxygenase-2-deficient beans and
those having a modified sugar profile, or the like. A process to
reduce oxygen levels in the formulation may also reduce the odor.
Various masking agents or fragrances may also be used to mask the
odor.
[0028] In yet another embodiment of the invention, the
soy-containing compositions may optionally comprise additional
synthetic or natural anti-cancer agents. Examples of such agents
include, without limitation, caffeine, Milk Thistle extract, green
tea extract, epigallocathechin gallate, silymarins, glucocorticoids
and 5-fluorouracil.
[0029] A preferred embodiment of the invention comprises the
administration of soymilk containing compositions before or after
the initiation of UV-induced skin cancer. Especially preferred are
embodiments in which the soymilk is not denatured, leaving STI and
BBI intact. Soymilk also contains genistein and other isoflavones,
and anti-oxidants such as the gamma form of vitamin E, which is
essential to the health of the skin. While not wishing to be held
to any particular theory, it is hypothesized that these different
active components also participate in the prevention of tumor
progression. Soymilk also contains lecithins and other emulsifying
molecules that facilitate the transdermal delivery of the active
components.
[0030] As explained above, the present invention extends to a
topical cosmetic or pharmaceutical composition comprising a
non-denatured soy product (e.g., a non-denatured soymilk or soybean
powder) and a cosmetic or pharmaceutically acceptable vehicle and,
optionally, additional anti-cancer or cosmetically active agents.
As used herein, "topically applying" means directly laying on or
spreading on outer skin, e.g., by use of the hands or an applicator
such as a wipe, roller, or spray.
[0031] The phrase "cosmetic or pharmaceutically acceptable" refers
to entities and compositions that are physiologically tolerable and
do not typically produce an allergic or similar untoward reaction
when administered to a human. As used herein, "cosmetically
acceptable" means that the ingredients which the term describes are
suitable for use in contact with tissues (e.g., the skin) without
undue toxicity, incompatibility, instability, irritation, allergic
response, and the like.
[0032] The term "vehicle" refers to a diluent, adjuvant, excipient,
or carrier. Such cosmetic or pharmaceutical vehicles can be
liquids, such as water and oils, including those of petroleum,
animal, vegetable or synthetic origin, such as peanut oil, soybean
oil, mineral oil, sesame oil and the like. In the art of
formulating skin care compositions, the vehicle is often an
oil-in-water or a water-in-oil emulsion. Suitable pharmaceutical
carriers are described in "Remington's Pharmaceutical Sciences" by
E. W. Martin. Suitable cosmetic carriers are described below.
[0033] The compositions for use in the methods of the present
invention include formulations suitable for topical application to
skin. In one embodiment, the composition comprises a non-denatured
soy product and a cosmetically acceptable topical carrier. In one
embodiment, the cosmetically acceptable topical carrier is from
about 50% to about 99.99%, by weight, of the composition (e.g.,
from about 80% to about 99%, by weight, of the composition).
[0034] The compositions may be made into a, wide variety of product
types that include, but are not limited to, solutions, lotions,
creams, gels, sticks, sprays, ointments, cleansing liquid washes,
solid bars, shampoos, pastes, foams, powders, mousses, shaving
creams, wipes, patches, nail lacquers, wound dressing, adhesive
bandages, hydrogels, and films. Make-up, such as foundations,
mascaras, and lipsticks also form suitable compositions. These
product types may comprise several types of cosmetically acceptable
topical carriers including, but not limited to solutions, emulsions
(e.g., microemulsions and nanoemulsions), gels, solids and
liposomes. Certain non-limitative examples of such carriers are set
forth hereinbelow. Other suitable carriers may be formulated by
those of ordinary skill in the art.
[0035] Topical compositions useful in the subject invention may be
formulated as a solution comprising an emollient. Such compositions
preferably contain from about 1% to about 50% of an emollient(s).
As used herein, the term "emollient" refers to materials used for
the prevention or relief of dryness, as well as for the protection
of the skin. A wide variety of suitable emollients is known and may
be used in the present invention. Sagarin, Cosmetics, Science and
Technology, 2nd Edition, Vol. 1, pp. 32-43 (1972) and the
International Cosmetic Ingredient Dictionary and Handbook, eds.
Wenninger and McEwen, pp. 1656-61, 1626, and 1654-55 (The Cosmetic,
Toiletry, and Fragrance Assoc., Washington, D.C., 7.sup.th Edition,
1997) (hereinafter "ICI Handbook") contains numerous examples of
suitable materials.
[0036] A lotion can be made from such a solution. Lotions typically
comprise from about 1% to about 20% (e.g., from about 5% to about
10%) of an emollient(s) and from about 50% to about 90% (e.g., from
about 60% to about 80%) of water.
[0037] Another type of product that may be formulated from a
solution is a cream. A cream typically comprises from about 5% to
about 50% (e.g., from about 10% to about 20%) of an emollient(s)
and from about 45% to about 85% (e.g., from about 50% to about 75%)
of water.
[0038] Yet another type of product that may be formulated from a
solution is an ointment. An ointment may comprise a simple base of
animal or vegetable oils or semi-solid hydrocarbons. An ointment
may comprise from about 2% to about 10% of an emollient(s) plus
from about 0.1% to about 2% of a thickening agent(s). A more
complete disclosure of thickening agents or viscosity increasing
agents useful herein can be found in Sagarin, Cosmetics, Science
and Technology, 2nd Edition, Vol. 1, pp. 72-73 (1972) and the ICI
Handbook pp. 1693-1697.
[0039] The topical compositions useful in the present invention may
be formulated as emulsions. If the carrier is an emulsion, from
about 1% to about 10% (e.g., from about 2% to about 5%) of the
carrier comprises an emulsifier(s). Emulsifiers may be nonionic,
anionic or cationic. Suitable emulsifiers are disclosed in, for
example, in McCutcheon's Detergents and Emulsifiers, North American
Edition, pp. 317-324 (1986), and the ICI Handbook, pp.
1673-1686.
[0040] Lotions and creams can be formulated as emulsions. Typically
such lotions comprise from 0.5% to about 5% of an emulsifier(s).
Such creams would typically comprise from about 1% to about 20%
(e.g., from about 5% to about 10%) of an emollient(s); from about
20% to about 80% (e.g., from 30% to about 70%) of water; and from
about 1% to about 10% (e.g., from about 2% to about 5%) of an
emulsifier(s).
[0041] Single emulsion skin care preparations, such as lotions and
creams, of the oil-in-water type and water-in-oil type are well
known in the cosmetic art and are useful in the present invention.
Multiphase emulsion compositions, for example the
water-in-oil-in-water type, as disclosed in U.S. Pat. Nos.
4,254,105 and 4,960,764, may also be useful in the present
invention. In general, such single or multiphase emulsions contain
water, emollients, and emulsifiers as essential ingredients.
[0042] The topical compositions of this invention can also be
formulated as a gel (e.g., an aqueous, alcohol, alcohol/water, or
oil gel using a suitable gelling agent(s)). Suitable gelling agents
for aqueous gels include, but are not limited to, natural gums,
acrylic acid and acrylate polymers and copolymers, and cellulose
derivatives (e.g., hydroxymethyl cellulose and hydroxypropyl
cellulose). Suitable gelling agents for oils (such as mineral oil)
include, but are not limited to, hydrogenated
butylene/ethylene/styrene copolymer and hydrogenated
ethylene/propylene/styrene copolymer. Such gels typically comprise
between about 0.1% and 5%, by weight, of such gelling agents.
[0043] The topical compositions of the present invention can also
be formulated as a solid formulation (e.g., a wax-based stick, soap
bar composition, powder, or a wipe containing powder).
[0044] Liposomal formulations are also useful compositions of the
subject invention. In one embodiment, the soymilk or soybean powder
particles or soy proteins such as STI are contained within the
liposome. Examples of liposomes are unilamellar, multilamellar, and
paucilamellar liposomes, which may or may not contain
phospholipids. Such compositions can be prepared by first combining
the non-denatured soy milk product or the STI with a phospholipid,
such as dipalmitoylphosphatidyl choline, cholesterol and water. An
example of a method for producing liposomes is described in Mezei
& Gulasekharam, "Liposomes--A Selective Drug Delivery System
for the Topical Route of Administration; Gel Dosage Form", Journal
of Pharmaceutics and Pharmacology, Vol. 34 (1982), pp. 473-474.
Those of skill in the art may make suitable modifications of the
method described therein.
[0045] Epidermal lipids of suitable composition for forming
liposomes may be substituted for the phospholipid. The liposome
preparation may then be incorporated into one of the above carriers
(e.g., a gel or an oil-in-water emulsion) in order to produce the
liposomal formulation. Other compositions and uses of topically
applied liposomes are described in Mezei, M., "Liposomes as a Skin
Drug Delivery System", Topics in Pharmaceutical Sciences (D.
Breimer and P. Speiser, eds.), Elsevier Science Publishers B. V.,
New York, N.Y., 1985, pp. 345-358, PCT Patent Application No.
WO96/31194, Niemiec, et al., 12 Pharm. Res. 1184-88 (1995), and
U.S. Pat. No. 5,260,065.
[0046] In one embodiment, the liposome is nonionic. In one example,
the liposome contains (a) glycerol dilaurate; (b) compounds having
the steroid backbone found in cholesterol; and (c) fatty acid
ethers having from about 12 to about 18 carbon atoms. In a further
embodiment, the liposome comprises glycerol dilaurate, cholesterol,
polyoxyethylene-10-stearyl ether, and polyoxyethylene-9-lauryl
ether. In one embodiment, these ingredients are in a ratio of about
38:12:33:17.
[0047] In one embodiment, the liposomes are present in the topical
composition in an amount, based upon the total volume of the
composition, of from about 5 mg/ml to about 100 mg/ml such as from
about 10 mg/ml to about 50 mg/ml.
[0048] The topical compositions useful in the subject invention may
contain, in addition to the aforementioned components, a wide
variety of additional oil-soluble materials and/or water-soluble
materials conventionally used in compositions for use on skin,
hair, and nails at their art-established levels.
[0049] In addition to such agents, other emollients and surface
active agents can be incorporated in the emulsions, including
glycerol trioleate, acetylated sucrose distearate, sorbitan
trioleate, polyoxyethylene (1) monostearate, glycerol monooleate,
sucrose distearate, polyethylene glycol (50) monostearate,
octylphenoxypoly (ethyleneoxy) ethanol, decaglycerin
penta-isostearate, sorbitan sesquioleate, hydroxylated lanolin,
lanolin, triglyceryl diisostearate, polyoxyethylene (2) oleyl
ether, calcium stearoyl-2-lactylate, methyl glucoside
sesquistearate, sorbitan monopalmitate, methoxy polyethylene
glycol-22/dodecyl glycol copolymer (Elfacos E200), polyethylene
glycol-45/dodecyl glycol copolymer (Elfacos ST9), polyethylene
glycol 400 distearate, and lanolin derived sterol extracts, glycol
stearate and glycerol stearate; alcohols, such as cetyl alcohol and
lanolin alcohol; myristates, such as isopropyl myristate; cetyl
palmitate; cholesterol; stearic acid; propylene glycol; glycerine,
sorbitol and the like.
[0050] The pharmaceutical or cosmetic composition may be optionally
combined with other ingredients such as moisturizers, cosmetic
adjuvants, anti-oxidants, depigmenting agents, darkening agents,
anti-aging agents, hair removal agents, hair styling agents, nail
styling agents, sunscreens, surfactants, bleaching agents, foaming
agents, conditioners, humectants, fragrances, colorants,
viscosifiers, buffering agents, preservatives, and the like and
mixtures thereof. Skin-care compositions including these components
should be formulated so as not to affect the soy product or soy
trypsin inhibitory activity.
[0051] Examples of humectants include glycerol, sorbitol, propylene
glycol, ethylene glycol, 1,3-butylene glycol, polypropylene glycol,
xylitol, malitol, lactitol, allantoin, acetamine MEA, oat protein,
hyaluronic acid, and the like. They may be used either singly or in
combination.
[0052] Because the compositions of this invention are
non-denatured, i.e., compositions in which the protease inhibitory
activity is retained, they may be more favorable as a medium for
microbial growth. Preservatives are useful for substantially
preventing microbial decomposition. Examples of preservatives
include phenoxyethanol and parabens such as methyl-paraben,
ethyl-paraben, and propyl-paraben; salicylic acid, chlorhexidine
hydrochloride, phenoxyethanol, sodium benzoate, methyl
para-hydroxybenzoate, ethyl para-hydroxybenzoate, propyl
para-hydroxybenzoate,.butyl para-hydroxybenzoate, isothiazolones
and the like. Other examples of preservatives are listed on pages
1654-55 of the International Cosmetic Ingredient Dictionary and
Handbook, eds. Wenninger and McEwen (CTFA, 7.sup.th ed., 1997),
hereinafter referred to as the "Cosmetic Handbook." The composition
may-comprise from about 0.01% to about 20%, by weight (more
preferably, from about 0.5% to about 5%, by weight) of
preservative. Microbial contamination can also be eliminated by
gamma irradiation or microfiltration, or by brief heat treatments
that do not result in the elimination of protease inhibitory
activity.
[0053] Examples of fragrances and odor masks include menthol,
anethole, carvone, eugenol, limonene, ocimene, n-decylalcohol,
citronellol, a-terpineol, methyl salicylate, methyl acetate,
citronellyl acetate, cineole, linalool, ethyl linalool, vanillin,
thymol, spearmint oil, peppermint oil, lemon oil, orange oil, sage
oil, rosemary oil, cinnamon oil, pimento oil, cinnamon leaf oil,
perilla oil, wintergreen oil, clove oil, eucalyptus oil and the
like.
[0054] Examples of surface active agents include sodium alkyl
sulfates, e.g., sodium lauryl sulfate and sodium myristyl sulfate,
sodium N-acyl sarcosinates; e.g., sodium N-lauroyl sarcosinate and
sodium N-myristoyl sarcosinate, sodium dodecylbenzenesulfonate,
sodium hydrogenated coconut fatty acid monoglyceride sulfate,
sodium lauryl sulfoacetate and N-acyl glutamates, e.g., N-palmitoyl
glutamate, N-methylacyltaurin sodium salt, N-methylacylalanine
sodium salt, sodium .alpha.-olefin sulfonate and sodium
dioctylsulfosuccinate; N-alkylaminoglycerols, e.g.,
N-lauryldiaminoethylglycerol and N-myristyldiaminoethylglycerol,
N-alkyl-N-carboxymethylammonium betaine and sodium
2-alkyl-l-hydroxyethylimidazoline betaine; polyoxyethylenealkyl
ether, polyoxyethylenealkylaryl ether, polyoxyethylenelanolin
alcohol, polyoxyethyleneglyceryl monoaliphatic acid ester,
polyoxyethylenesorbitol aliphatic acid ester, polyoxyethylene
aliphatic acid ester, higher aliphatic acid glycerol ester,
sorbitan aliphatic acid ester, Pluronic.TM. type surface active
agent, and polyoxyethylenesorbitan aliphatic acid esters such as
polyoxyethylenesorbitan monooleate and polyoxyethylenesorbitan
monolaurate.
[0055] Examples of the binder or thickener include cellulose
derivatives such as alkali metal salts of carboxymethylcellulose,
methyl cellulose, hydroxyethyl cellulose and sodium
carboxymethylhydroxyethyl cellulose, alkali metal alginates such as
sodium alginate, propylene glycol alginate, gums such as
carrageenan, xanthan gum, tragacanth gum, caraya gum and gum
arabic, and synthetic binders such as polyvinyl alcohol, polysodium
acrylate and polyvinyl pyrrolidone. Thickening agents that can be
added to the compositions of this invention to alter viscosity
include other polymers such as polyacrylates (e.g.,
polyacrylamide). Other examples of viscosity modifying agents are
listed on pages 1692-97 of the Cosmetic Handbook. To achieve the
appropriate viscosity, compositions of the present invention may
comprise from about 0.01% to about 20%, by weight (e.g., from about
0.1% to about 5%, by weight) of a thickening agent.
[0056] Coloring agents and fragrances also are commonly included in
such compositions.
[0057] In one embodiment, the topical composition further comprises
another cosmetically active agent in addition to the non-denatured
soy product. A "cosmetically active agent" is a compound (e.g., a
synthetic compound or a compound isolated from a natural source or
a natural extract) that has a cosmetic or therapeutic effect on the
skin, hair, or nails, including, but not limiting to, lightening
agents, darkening agents such as self-tanning agents, anti-acne
agents, shine control agents, anti-microbial agents,
anti-inflammatory agents, anti-mycotic agents, anti-parasite
agents, external analgesics, sunscreens, photoprotectors,
antioxidants, keratolytic agents, detergents/surfactants,
moisturizers, nutrients, vitamins, energy enhancers,
anti-perspiration agents, astringents, deodorants, hair removers,
firming agents, anti-callous agents, and agents for hair, nail,
and/or skin conditioning.
[0058] The compositions of this invention may be applied prior to,
concurrently with or after other active ingredients or compositions
to enhance their effect.
[0059] Antioxidants and/or chelating agents may also be used to
increase shelf life and stability of the compositions. Antioxidants
may be added both for formulation stabilization and for biological
efficacy. Antioxidant compounds and their derivatives include, but
are not limited to, water-soluble antioxidants such as sulfhydryl
compounds and their derivatives (e.g., sodium metabisulfite and
N-acetyl-cysteine), lipoic acid and dihydrolipoic acid,
resveratrol, acetyl-cysteine (Iniferine.RTM.) or lactoferrin, and
ascorbic acid and ascorbic acid derivatives (e.g., ascorbyl
palmitate and ascorbyl polypeptide). Oil-soluble antioxidants
suitable for use in the compositions of this invention include, but
are not limited to, butylated hydroxytoluene, retinoids (e.g.,
retinol and retinyl palmitate), tocopherols (e.g., tocopherol
acetate), tocotrienols, and ubiquinone. Natural extracts containing
antioxidants suitable for use in the compositions of this
invention, include, but not limited to, extracts containing
flavonoids and isoflavonoids and-their derivatives (e.g., genistein
and diadzein), extracts containing resveratrol and the like.
Examples of such natural extracts include grape seed, green tea,
pine bark, propolis, and legume extracts. Other examples of
antioxidants may be found on pages 1612-13 of the Cosmetic
Handbook. The compositions of the present invention may comprises
the antioxidant in an amount of from about 0.001% to about 20%, by
weight (e.g., from about 0.01% to about 10% by weight) of the
composition.
[0060] It is preferable to have at least one oil-soluble
antioxidant in the compositions of this invention. The antioxidants
should be utilized in a stabilizing effective amount and may range
in total from about 0.001 to 10% based on the weight of the total
composition, preferably from about 0.005 to about 5%. The
oil-soluble antioxidants which are useful in the compositions of
the present invention include butylated hydroxytoluene (BHT),
ascorbyl palmitate, butylated hydroxanisole (BHA),
phenyl-.alpha.-naphthylamine, hydroquinone, propyl gallate,
nordihydroguiaretic acid, and mixtures thereof as well as any other
known oil-soluble antioxidant compatible with the other components
of the compositions.
[0061] Preferably, a water-soluble antioxidant should also be
present in the water phase of the compositions of this invention.
The water-soluble antioxidants which are useful in the compositions
of this invention include ascorbic acid, sodium metabisulfite,
sodium bisulfite, sodium thiosulfite, sodium formaldehyde
sulfoxylate, isoascorbic acid, thioglyerol, thiosorbitol, thiourea,
thioglycolic acid, cysteine hydrochloride,
1,4-diazobicyclo-(2,2,2)-octane and mixtures thereof as well as any
other known water-soluble antioxidant compatible with the other
components of the compositions.
[0062] Chelating agents are also useful in assisting the
stabilization of the compositions of this invention. Examples of
chelating agents include EDTA and derivatives thereof (e.g.,
disodium EDTA and dipotassium EDTA), Iniferine.RTM., lactoferrin,
and citric acid. Other examples of chelating agents are listed on
page 1626 of the Cosmetic Handbook. The compositions of the present
invention may comprise the chelating agent in an amount of from
about 0.001% to about 20%, by weight (e.g., from about 0.01% to
about 10% by weight) of the composition.
[0063] Other active ingredients such as sunscreen materials may be
utilized in the compositions of the present invention provided that
they are physically and chemically compatible with the other
components of the compositions. Sunscreens may include organic or
inorganic sunscreens, such as methoxyoctylcinnamate and other
cinnamate compounds, titanium dioxide and zinc oxide and the
like.
[0064] Various irritancy mitigants may be added to the compositions
of this invention. Irritancy mitigants such as .alpha.-bisabolol,
panthenol, allantoin, ginkgo biloba, stearoyl glycerrhetinic acid
(licorice extract), tea tree oil, butchers' broom, calendula,
ginseng and the like may be added.
[0065] Other ingredients may include agents that assist in
protecting the skin from aging, such as sunscreens, anti-oxidant
vitamins such as ascorbic acid, vitamin B, biotin, pantothenic
acid, vitamin D, vitamin E and vitamin C, and sodium bisulfite.
Yeast extract, gingko biloba, bisabolol, panthenol, alpha hydroxy
acids and oligosaccharides such as melibiose are among other
ingredients which assist in preventing aging of the skin by such
means as irritation mitigation, oxidation mitigation, healing,
affecting retinoid metabolism and inhibiting the production of
elastase.
[0066] The compositions of this invention may also contain other
depigmenting agents in addition to the soy product. What is meant
by depigmentation is the lightening of the color of an area of
skin, including but not limited to, the global lightening of the
user's skin tone/complexion (e.g., the face, hands, or whole body,
which is uneven as a result of aging skin, or darker than desired
because of ethnicity or pathology, and the like), the evening of
skin color tone, or the specific lightening of age spots, freckles,
or darker pigmented areas such as, but not limited to,
post-inflammatory hyper-pigmentary lesions.
[0067] Examples of such depigmenting agents include, but are not
limited to, lipoic acid, dihydrolipoic acid, resveratrol, ascorbic
acid, kojic acid, hydroquinone, isoflavones, retinoids (e.g.,
retinol, retinoic acid, and retinyl palmitate), tyrosinase
inhibitors, melanosome transfer inhibitors, and selective cytotoxic
agents for melanocytes, or natural extracts, e.g., licorice
extract, gatuline A (pilewort extract), and micromerol (butylene
glycol and apple extract), providing these activities. The amount
of the depigmenting agent used will depend on the activity of the
compound, and will typically range from about 0.001% to about 20%,
by weight (e.g., from about 0.01% to about 10%, by weight) of the
composition.
[0068] Other skin color evening ingredients, such as skin darkening
or sunless tanning agents, may also be effective in the skin care
compositions for use in this invention.
[0069] The composition of the present invention may also contain
compounds that enhance the feel of the composition on the skin of
the user. Examples of such compounds include, but are not limited
to, oils, silicones (e.g., siloxane polymers such as dimethicone)
and skin-conditioning agents such as emollients, and humectants.
Examples of such skin conditioning agents may be found of pages
1656-1670 of the Cosmetic Handbook.
[0070] Compositions which assist in the reduction of lines and
wrinkles may also be added to the compositions of this invention.
For example, alpha hydroxy acids, hyaluronic acid, Gatuline R
(fagus silvitica extract), pigments and scattering aids such as
zinc oxide and titanium dioxide may be used in the compositions of
this invention in this capacity.
[0071] Anti-inflammatory agents may also be used in the
compositions of this invention. Not only should these agents assist
in mitigating irritation, they may assist in treating wrinkles and
lines in the skin. Steroidal anti-inflammatory agents, including
but not limited to, corticosteroids such as hydrocortisone,
hydroxyltriamcinolone, alpha-methyl dexamethasone,
dexamethasone-phosphate, beclomethasone dipropionate, clobetasol
valerate, desonide, desoxycorticosterone acetate, dexamethoasone,
dichlorisone, deflorasonediacetate, diflucortolone valerate,
fluadronolone, fluclarolone acetonide, fludrocortisone,
flumethasone pivalate, fluosinolone acetonide, fluocionide,
flucortine butylester, fluocortolone, flupredidene (flupredylidene)
acetate, flurandronolone, halcinonide, hydrocortisone acetate,
hydrocortisone butyrate, methylprednisolone, triamcinolone
acetonide, cortisone, cortodoxone, flucetonide, fludrocortisone,
difluorosone diacetate, fluradrenalone acetonide, medrysone,
amciafel, amcinafide, betamethasone and its esters, chlorprednisone
acetate, clocortelone, clescinolone, dichlorisone, difluprednate,
flucloronide, flunisolide, fluoromethalone, fluperolone,
fluprednisolone, hydrocortisone valerate, hydrocortisone
cyclopentylpropionate, hydrocortamate, meprednisone, paramethasone,
prednisolone, prednisone, beclomethasone dipropionate,
triamcinolone and mixtures thereof may be used. Preferably,
hydrocortisone or natural extracts with similar activity may be
used.
[0072] Nonsteroidal anti-inflammatory agents may also be employed
in the compositions of this invention, such as salicylates, acetic
acid derivatives, fenamates, propionic acid derivatives and
pyrazoles or mixtures thereof. Other synthetic and natural
anti-inflammatory agents may also be used.
[0073] Additional active ingredients having topical activity may be
utilized in the compositions of this invention. Azole-type
anti-fungal and anti-bacterial agents may be employed in the
compositions of this invention in their base form. For example,
ketoconazole, miconazole, itraconazole, elubiol, and like related
imidazole antifungals and antibacterials are useful in the topical
formulations of this invention.
[0074] It can be readily appreciated that a transdermal route of
administration may be enhanced by use of a dermal penetration
enhancer, e.g., such as enhancers described in U.S. Pat. No.
5,164,189, U.S. Pat. No. 5,008,110, and U.S. Pat. No. 4,879,119,
issued Nov. 7, 1989 to Aruga et al. In one embodiment, a
composition of the present invention can be delivered in a
controlled release system, such as using a transdermal patch,
liposomes, or other modes of administration. In another embodiment,
polymeric materials can be used [see Medical Applications of
Controlled Release, Langer and Wise (eds.), CRC Press: Boca Raton,
Fla. (1974); Controlled Drug Bioavailability, Drug Product Design
and Performance, Smolen and Ball (eds.), Wiley: New York (1984);
Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23:61
(1983); see also Levy et al., Science 228:190 (1985); During et
al., Ann. Neurol. 25:351 (1989); Howard et al., J. Neurosurg.
71:105 (1989)].
[0075] In another embodiment, a controlled release system can be
placed in proximity of the target tissues of the mammal, thus
requiring only a fraction of the systemic dose [see, e.g., Goodson,
in Medical Applications of Controlled Release, supra, vol. 2, pp.
115-138 (1984)]. In particular, a controlled release system can be
introduced into an animal in proximity of the site of inappropriate
immune activation or a tumor. Other controlled release systems are
discussed in a review by Langer [Science 249:1527-1533 (1990)].
[0076] In yet another embodiment of the invention, the soybean
trypsin inhibitor may be produced by recombinant means. The
nucleotide and protein sequences of STI are known. See GenBank
Accession No. AF314823. Methods for recombinant expression of STI
are well known to those of ordinary skill in the art. In an
alternative embodiment, the STI so produced may be modified at the
genetic level (e.g. replacing amino acids to change local charges,
to enhance skin penetration without compromising activity, or to
enhance activity without compromising skin penetration) or
chemically post synthesis (e.g. additional lipid or sugar groups)
to enhance uptake of the STI into the skin of the patient.
[0077] Various and numerous methods are known in the art for
transdermal administration of a drug, e.g., via a transdermal
patch. Transdermal patches are described in for example, U.S. Pat.
No. 5,407,713, issued Apr. 18, 1995 to Rolando et al.; U.S. Pat.
No. 5,352,456, issued Oct. 4, 1004 to Fallon et al.; U.S. Pat. No.
5,332,213 issued Aug. 9, 1994 to D'Angelo et al.; U.S. Pat. No.
5,336,168, issued Aug. 9, 1994 to Sibalis; U.S. Pat. No. 5,290,561,
issued Mar. 1, 1994 to Farhadieh et al.; U.S. Pat. No. 5,254,346,
issued Oct. 19, 1993 to Tucker et al.; U.S. Pat. No. 5,164,189,
issued Nov. 17, 1992 to Berger et al.; U.S. Pat. No. 5,163,899,
issued Nov. 17, 1992 to Sibalis; U.S. Pat. Nos. 5,088,977 and
5,087,240, both issued Feb. 18, 1992 to Sibalis; U.S. Pat. No.
5,008,110, issued Apr. 16, 1991 to Benecke et al.; and U.S. Pat.
No. 4,921,475, issued May 1, 1990 to Sibalis.
[0078] Compositions of the present invention may be prepared by
mixing the desired ingredients. For example, soymilk is mixed with
the chelating agent, preservative, and/or antioxidant. A thickener
is then added to the system, and the mixture is further mixed until
it reaches homogeneity at the desired viscosity. The compositions
of the present invention may be prepared under an argon, nitrogen,
or other inert gaseous blanket in order to enhance formulation
stability and/or to reduce soybean odor. The compositions of this
invention may be packaged in a tube, a sealed packet, a jar, a
pump, a bottle, a can, a pledget, a towelet, a wipe or the like. An
airtight package such as an aluminum tube, aluminum pocket, pump,
laminate tube, or the like, can also be used to further enhance
product stability.
[0079] The skin-care compositions for use in the methods of this
invention may be applied daily for at least four weeks, and more
preferably at least eight weeks, by which an effect upon the
appearance of skin should be observed. Application may be continued
as long as desired to maintain the condition of the skin and to
reduce skin cancer risk.
[0080] The topically active pharmaceutical or cosmetic composition
should be applied in an amount effective to effect the desired
changes in the skin. As used herein, "amount effective" shall mean
an amount sufficient to cover the region of skin surface where
preventing cancer, inhibiting the growth rate of a cutaneous tumor,
or reducing the risk of cancer is desired. Preferably, the
composition is applied to the skin surface such that, based upon a
cm.sup.2 of skin surface, from about 2 .mu.l/cm.sup.2 to about 500
.mu.l/cm.sup.2 of topically active agent is present when preventing
cancer, inhibiting the growth rate of a cutaneous tumor, or
reducing the risk of cancer is desired.
[0081] The following examples are provided to describe the
invention in further detail. These examples are provided for
illustrative purposes only, and-are not to be construed as limiting
the invention.
EXAMPLE 1
Soymilk Reduces UVB-Induced Tumorigenesis
[0082] Female SKH-1 mice, 6-7 weeks old, were purchased from
Charles River Laboratories (Kingston, N.Y.) and were housed for at
least one week before use. Mice were given water and Purina
Laboratory Chow 5001 diet (Ralston-Purina, St. Louis, Mo.) ad
libitum. Animal care was based on the "Guide for the Care and Use
of Laboratory Animals", NIH Publication No. 85-23. Ultraviolet B
light (UVB) lamps (FS72T12-UVB-HO) were from National Biological
Corp. (Twinsburg, Ohio). Mice were irradiated with UV lamps that
emit both UVB (280-320 nm, 75-80% of total energy) and UVA (320-375
nm, 20-25% of total energy). The UVB dose was quantified with a
Spectra 305 dosimeter from DaevIin Co. (Byran, Ohio). The radiation
was further calibrated with a Research radiometer/photometer model
IL1700 from International Light Inc. (Newburgport, Mass.).
[0083] Mice were irradiated with UVB light (30 mJ/cm.sup.2) for 25
to 30 seconds twice a week for 20 weeks. UVB irradiation was
stopped when the first tumor was visible on the back of one mouse.
This mouse was removed, and the population was classified as "at
high risk for skin tumor development." Three weeks later, mice with
no tumors were randomized into groups of 30 mice. One group (the
control) was not treated, and other groups were topically treated,
once a day, five days a week, with 100 .mu.l of test material.
Agents tested in this example included STI and BBI, and either
freshly prepared soymilk or freshly prepared soymilk that was heat
denatured at 90.sup.EC for 20 min. Controls include vehicle alone
and an unrelated protein (BSA).
[0084] The number of skin tumors and tumor sizes were measured
every three weeks, using the methods described in Lou et al.,
(Effects of oral administration of tea, decaffeinated tea, and
caffeine on the formation and growth of tumors in high risk SKH-1
mice previously treated with ultraviolet B light. Nutrition and
Cancer 33, p. 146-153, 1999). The results of these measurements are
summarized in Table 1 and shown in FIGS. 1A-1C and FIGS. 2A and 2B.
An increase in tumor volume per mouse during the 21 weeks of the
treatment phase was observed in each treatment regimen. The daily
treatment with heated soymilk did not affect the rate of increase
in tumor volume per mouse relative to control. Surprisingly, daily
treatment with fresh soymilk had a dramatic effect on tumor volume.
Tumor growth was significantly slower, and final tumor volumes were
reduced by a factor of two to three in the mice treated with fresh
soymilk, relative to the untreated or heated soymilk treated mice.
See FIG. 2C. Table 1 shows the effect of fresh soymilk on tumor
progression as measured by several additional parameters. The
percent of tumor-bearing mice was reduced following treatment with
fresh soymilk, as were the number of tumors per mouse and the tumor
volume per tumor.
[0085] FIGS. 1A-1C show the effects of treatment of BBI and STI on
mice irradiated with UVB. Surprisingly, using STI alone had an
unexpected and significant effect on tumor progression. See FIGS.
1A-1C. The dramatic difference in results produced by treatment
with fresh soymilk versus heat denatured soymilk, see FIGS. 2A-2C,
also points to STI as an active anti-tumor ingredient, since it is
well known that STI is denatured by cooking, but heat alone will
not denature BBI, nor does it affect phytoestrogens. FIGS. 4A-4C
are three photographs showing a random group of mice from each
treatment group. This figure clearly demonstrates the reduced
tumorigenicity in the mice treated with fresh soymilk vs heated
treated soymilk or water. Compare FIGS. 4A with 4B or 4C. These
data indicate that topical treatment with fresh soymilk reduces
UVB-induced tumor growth and progression in a high risk,
pre-exposed population. These data also suggest that some of the
active(s) of fresh soymilk that are involved in the tumor growth
reduction are heat sensitive.
1TABLE I Effects of Topical Applications of Soymilk, STI, BBI, and
Caffeine on the Development of Skin Tumors in SKH-1 Mice Previously
Treated with Ultraviolet B Light Number of Percent of Tumor Tumor
mouse per Weight per mice with Tumors per volume per volume per
Week Treatment group mouse (g) tumors mouse rumor (mm.sup.3) mouse
(mm.sup.3) 0 No treatment 30/30 29.9 .+-. 0.5 0 0 0 0 Water 29/29
30.0 .+-. 0.3 0 0 0 0 Soymilk 30/30 29.9 .+-. 0.5 0 0 0 0 Heated
Soymilk 30/30 30.0 .+-. 0.5 0 0 0 0 Liposomes 30/30 30.0 .+-. 0.4 0
0 0 0 STI 30/30 30.2 .+-. 0.4 0 0 0 0 BBI 30/30 30.1 .+-. 0.4 0 0 0
0 BSA in 30/30 30.1 .+-. 0.2 0 0 0 0 liposomes Acetone 30/30 30.1
.+-. 0.2 0 0 0 0 Caffeine 30/30 30.0 .+-. 0.2 0 0 0 0 (1.2 mg) 6 No
treatment 30/30 30.3 .+-. 0.4 47.7% 0.7 .+-. 0.2 1.5 .+-. 0.7 1.1
.+-. 0.5 Water 29/29 30.4 .+-. 0.5 27.6% 0.4 .+-. 0.2 0.5 .+-. 0.0
0.2 .+-. 0.1 Soymilk 30/30 29.9 .+-. 0.4 6.7% 0.1 .+-. 0.0 0.5 .+-.
0.0 0 Heated Soymilk 30/30 30.4 .+-. 0.6 36.7% 0.4 .+-. 0.1 1.1
.+-. 0.4 0.5 .+-. 0.2 Liposomes 30/30 30.0 .+-. 0.4 40.0% 0.7 .+-.
0.2 2.0 .+-. 0.7 1.4 .+-. 0.6 STI 30/30 29.7 .+-. 0.5 16.7% 0.2
.+-. 0.1 3.5 .+-. 1.9 0.8 .+-. 0.6 BBI 30/30 30.0 .+-. 0.5 30.0%
0.4 .+-. 0.1 0.9 .+-. 0.3 0.3 .+-. 0.1 BSA in 30/30 30.3 .+-. 0.4
20.0% 2.0 .+-. 0.1 2.5 .+-. 1.9 0.6 .+-. 0.5 liposomes Acetone
30/30 29.6 .+-. 0.4 36.7% 0.7 .+-. 0.2 2.6 .+-. 0.9 1.9 .+-. 0.8
30/30 29.3 .+-. 0.4 16.7% 0.2 .+-. 0.1 1.3 .+-. 0.7 0.2 .+-. 0.1 9
No treatment 30/30 30.5 .+-. 0.3 43.3% 0.8 .+-. 0.2 3.3 .+-. 1.5
2.7 .+-. 1.3 Water 29/29 30.6 .+-. 0.5 48.3% 1.0 .+-. 0.3 1.3 .+-.
0.3 1.3 .+-. 0.6 Soymilk 30/30 30.1 .+-. 0.4 20.0% 0.2 .+-. 0.1 0.5
.+-. 0.0 0.1 .+-. 0.0 Heated Soymilk 30/30 30.3 .+-. 0.5 46.7% 0.7
.+-. 0.1 4.8 .+-. 1.9 3.2 .+-. 1.3 Liposomes 30/30 30.2 .+-. 0.4
46.7% 1.9 .+-. 0.5 1.7 .+-. 0.6 3.2 .+-. 1.2 STI 29/30 30.0 .+-.
0.4 41.4% 0.9 .+-. 0.2 3.0 .+-. 1.4 2.6 .+-. 1.4 BBI 30/30 29.9
.+-. 0.5 27.6% 0.6 .+-. 0.2 1.7 .+-. 0.8 1.0 .+-. 0.7 BSA in 30/30
30.5 .+-. 0.4 50.0% 1.1 .+-. 0.3 2.7 .+-. 1.2 2.9 .+-. 1.4
liposomes Acetone 30/30 30.4 .+-. 0.5 60.0% 1.4 .+-. 0.5 2.8 .+-.
0.9 4.0 .+-. 1.5 Caffeine 30/30 29.4 .+-. 0.4 13.3% 0.2 .+-. 0.1
3.4 .+-. 2.2 0.7 .+-. 0.5 (1.2 mg) 12 No treatment 30/30 30.4 .+-.
0.4 70.0% 1.9 .+-. 0.3 2.8 .+-. 0.7 5.4 .+-. 1.7 Water 29/29 31.1
.+-. 0.5 58.6% 1.9 .+-. 0.5 2.3 .+-. 0.5 4.4 .+-. 1.2 Soymilk 30/30
30.4 .+-. 0.4 40.0% 0.7 .+-. 0.2 1.8 .+-. 0.4 1.2 .+-. 0.4 Heated
Soymilk 30/30 30.2 .+-. 0.6 53.3% 1.0 .+-. 0.2 14.4 .+-. 6.0 14.9
.+-. 6.3 Liposomes 30/30 30.0 .+-. 0.4 70.0% 4.0 .+-. 0.9 4.4 .+-.
1.0 17.8 .+-. 5.2 STI 29/30 29.8 .+-. 0.5 46.7% 1.9 .+-. 0.5 4.8
.+-. 1.3 9.4 .+-. 3.9 BBI 30/30 30.6 .+-. 0.4 50.0% 1.5 .+-. 0.4
2.4 .+-. 0.8 3.7 .+-. 1.8 BSA in 30/30 30.4 .+-. 0.5 80.0% 2.6 .+-.
0.6 3.5 .+-. 1.0 9.1 .+-. 3.1 liposomes Acetone 30/30 30.0 .+-. 0.4
80.0% 4.5 .+-. 0.9 5.2 .+-. 1.0 23.4 .+-. 7.1 Caffeine 30/30 29.0
.+-. 0.3 50.0% 1.3 .+-. 0.4 3.2 .+-. 0.8 4.0 .+-. 1.9 15 No
treatment 30/30 30.6 .+-. 0.4 76.7% 2.9 .+-. 0.5 5.8 .+-. 1.4 16.8
.+-. 5.7 Water 29/29 31.2 .+-. 0.4 82.8% 3.5 .+-. 0.6 2.8 .+-. 0.6
9.7 .+-. 3.4 Soymilk 30/30 30.7 .+-. 0.3 46.7% 1.1 .+-. 0.3 3.0
.+-. 0.8 3.4 .+-. 1.0 Heated Soymilk 30/30 30.6 .+-. 0.5 70,0% 2.3
.+-. 0.4 10.1 .+-. 3.6 22.9 .+-. 7.9 Liposomes 30/30 29.6 .+-. 0.9
90.0% 6.2 .+-. 1.1 7.5 .+-. 1.3 46.2 .+-. 13.2 STI 29/30 30.1 .+-.
0.4 55.2% 3.1 .+-. 0.7 5.8 .+-. 2.1 17.8 .+-. 7.9 BBI 30/30 30.6
.+-. 0.5 66.7% 2.2 .+-. 0.4 4.1 .+-. 0.8 9.0 .+-. 2.4 BSA in 30/30
30.5 .+-. 0.4 90.0% 5.3 .+-. 0.8 4.0 .+-. 0.7 21.4 .+-. 6.6
liposomes Acetone 30/30 30.0 .+-. 0.3 83.3% 6.6 .+-. 1.0 7.4 .+-.
1.3 48.3 .+-. 16.7 Caffeine 30/30 29.4 .+-. 0.3 56.7% 2.1 .+-. 0.5
3.2 .+-. 0.6 6.7 .+-. 2.2 (1.2 mg) 18 No treatment 30/30 31.6 .+-.
0.4 83.3% 4.2 .+-. 0.6 9.0 .+-. 2.0 38.0 .+-. 12.8 Water 28/29 32.0
.+-. 0.5 96.6% 5.4 .+-. 0.6 4.0 .+-. 1.3 21.5 .+-. 7.5 Soymilk
30/30 30.7 .+-. 0.4 66.7% 1.9 .+-. 0.5 6.5 .+-. 1.8 12.3 .+-. 4.6
Heated Soymilk 30/30 31.0 .+-. 0.6 83.3% 4.1 .+-. 0.7 6.9 .+-. 2.4
28.7 .+-. 9.8 Liposomes 30/30 30.9 .+-. 0.5 90.0% 7.1 .+-. 1.0 8.7
.+-. 1.1 61.7 .+-. 15.0 STI 28/30 30.0 .+-. 0.4 63.3% 4.2 .+-. 0.9
7.3 .+-. 2.2 30.7 .+-. 11.9 BBI 30/30 30.7 .+-. 0.5 76.7% 3.2 .+-.
0.6 5.9 .+-. 1.3 19.0 .+-. 4.6 BSA in 30/30 30.9 .+-. 0.5 90.0% 6.5
.+-. 0.9 6.3 .+-. 1.0 41.1 .+-. 9.7 liposomes Acetone 29/30 30.7
.+-. 0.5 86.7% 6.9 .+-. 1.0 8.4 .+-. 1.3 57.4 .+-. 13.7 Caffeine
30/30 29.6 .+-. 0.4 80.0% 3.6 .+-. 0.6 6.2 .+-. 1.3 22.1 .+-. 6.2
21 No treatment 30/30 31.5 .+-. 0.3 100.0% 5.9 .+-. 0.7 8.8 .+-.
2.1 51.9 .+-. 16.3 Water 28/29 32.0 .+-. 0.4 96.6% 6.3 .+-. 0.6 6.4
.+-. 2.4 40.6 .+-. 15.6 Soymilk 30/30 31.1 .+-. 0.3 83.3% 2.7 .+-.
0.5 4.8 .+-. 1.3 12.9 .+-. 4.4 Heated Soymilk 29/30 31.3 .+-. 0.6
90.0% 6.7 .+-. 0.9 8.9 .+-. 3.2 59.9 .+-. 20.9 Liposomes 30/30 30.9
.+-. 0.4 96.7% 8.2 .+-. 1.1 9.0 .+-. 1.5 73.7 .+-. 15.0 STI 28/30
30.6 .+-. 0.4 80.0% 5.6 .+-. 0.9 8.6 .+-. 3.2 48.7 .+-. 23.3 BBI
30/30 30.8 .+-. 0.4 90.0% 4.5 .+-. 0.6 7.4 .+-. 2.0 33.2 .+-. 9.6
BSA in 30/30 30.7 .+-. 0.4 90.0% 7.1 .+-. 0.8 6.3 .+-. 1.0 44.7
.+-. 11.6 liposomes Acetone 28/30 30.5 .+-. 0.4 90.0% 6.9 .+-. 0.9
9.6 .+-. 2.0 65.7 .+-. 0.9 Caffeine 30/30 29.9 .+-. 0.3 76.7% 4.0
.+-. 0.7 5.3 .+-. 1.0 21.5 .+-. 6.0 SKH-1 mice (7-8 weeks old) were
treated with ultraviolet B light (UVB; 30 mJ/cm.sup.2) twice weekly
for 20 weeks and UVB treatment was stopped. Three weeks later, the
mice (with no visible tumors) were randomized into 10 # groups (30
mice per group) and the mice were treated topically: group 2, 100
ul water; group 3, 100 ul Soymilk; group 4, 100 ul heated Soymilk;
group 5, 100 ul liposomes; # group 6, trypsin inhibitor (STI; 0.8
mg; T9003, Sigma) in 100 ul liposomes; group7, Bowman-Birk protease
inhibitor (BBI; 0.8 mg; T97770, Sigma) in 100 ul liposomes; group
8, bovine serum albumin (BSA; 0.8 mg) in 100 ul liposomes; # group
9, 100 ul acetone; group 10, caffeine, CAF; 1.2 mg) in 100 ul
acetone once a day 5 days per week for 21 weeks.
EXAMPLE 2
Caffeine Reduces UVB-Induces Tumorigenesis
[0086] The experiment described in Example 1 was also performed
using a topical caffeine treatment. Caffeine was purchased from
Sigma (St. Louis, Mo.). Acetone from Fisher Scientific
(Springfield, N.J.). Mice were treated daily, five days a week with
caffeine (1.2 mg) in 100 .mu.l acetone or with 100 .right
brkt-bot.l of acetone only. Experimental procedures and
measurements were identical to those described in Example 1, and
the two experiments were performed at the same time, using the same
UVB-irradiated population.
[0087] FIGS. 3A-3C show that tumor progression following acetone
treatment was very similar to that of the untreated group graphed
in FIG. 2. The caffeine treated mice showed a marked delay in tumor
progression, as evident from the tumor volume per mouse data in
FIG. 3C. The additional parameters reported in Table 1 further
support this observation of the beneficial effect of caffeine on
tumor progression. The percentage of tumor-bearing mice was reduced
following caffeine treatment, as were the number of tumors per
mouse and the tumor volume per tumor. FIGS. 5A and 5B are two
pictures, each depicting a random group of mice from acetone
treated (FIG. 5A) vs. caffeine treated (FIG. 5B mice). This figure
clearly demonstrates the reduced tumorigenicity in the
caffeine-treated mice.
EXAMPLE 3
Preparation of Soymilk from Soybean Powder
[0088] 160 g of soybean powder (Sunlight Foods, Taipei, Taiwan) was
added to about 1440 g of deionized water. The mixture was stirred
at room temperature for about 1 hour. The mixture was then filtered
through a sieve having holes of 75 .mu.m diameter. The filtrate
resulted in about 1.1 kg of soymilk.
EXAMPLE 4
Preparation of Soymilk Gel from Soymilk
[0089] The following compositions of this invention were prepared
as follows. The weight percentages of each ingredient in the
compositions are indicated below in Table 2 and Table 3. First, the
soymilk, as prepared in example 3, was placed into a first beaker.
The preservative Phenonip.RTM. (a mixture of the preservatives
methyl-paraben, propyl-paraben, ethyl-paraben, and phenoxy-ethanol
sold by NIPA, Wilmington, Del.) or the preservative phenoxyethanol
were added to the soymilk. Next, the chelating agent Disodium EDTA
and in some examples the humectant glycerin were added to the first
beaker and mixed with the soymilk. It is also possible to further
add cyclomethicone, or dimethicone (tradename Dow Corning 200
Fluid.RTM.), or PolySorbate 20, or Aluminum Starch Octyl Succinate,
or Sucrose Cocoate, or PEG-6 Capric/Caprylic Triglycerides to the
soymilk mixture at this step as required in some examples in Table
2 and Table 3. A mixture of the thickener polyacrylamide,
laureth-7, and C13-14 isoparaffins (sold by Seppic, Paris, France
under the tradename Sepigel.RTM.) was added to a second beaker
along with the anti-oxidant BHT. The ingredients in the second
beaker were then added to the ingredients of the first beaker and
mixed until homogenous. The anti-oxidants ascorbic acid, sodium
ascorbyl phosphate, lactoferrin, or tocopherol were then added to
the beaker and homogeneously mixed to form the resulting gel.
EXAMPLE 5
Preparation of Soymilk Gel from Soybean Powder, Soymilk Powder or
Soybean Extract
[0090] The following compositions of this invention were prepared
as follows. The weight percentage of each ingredient in each of the
preparations is indicated below in Table 3. First, the soymilk
powder (Devansoy Farms, Carroll, Iowa) or the soybean powder
(Sunlight Foods, Taipei, Taiwan) or the Soybean Extract and
deionized water were placed into a first beaker and mixed to
reconstitute the soy powder. The preservative Phenonip.RTM. and the
chelating agent Disodium EDTA were then added to the first beaker
and mixed with the soymilk. A mixture of polyacrylamide, laureth-7,
and C13-14 isoparaffins was added to a second beaker along with the
anti-oxidant BHT. The ingredients in the second beaker were then
added to the ingredients of the first beaker and mixed until
homogenous.
2TABLE 2 Soybean Essence formulations 24 26 27 28 33 34 35 Soymilk
94.40% 92.40% 90.70% 94.70% Phenoxyethanol and Parabens 1.00% 1.00%
1.00% 1.00% 1.00% 1.00% 1.00% Glycerin 5.00% Disodium EDTA 0.05%
0.05% 0.05% 0.05% 0.05% 0.05% 0.05%
Polyacrylamide/Laureth-7/C.sub.13-14 Isoparrafin 3.50% 3.50% 3.20%
3.20% 3.20% 3.20% 3.20% Ascorbic Acid 1.00% Butylated
Hydroxytoluene 0.05% 0.05% 0.05% 0.05% 0.05% 0.05% 0.05% Deionized
Water 90.70% 90.70% 85.70% Lactoferrin 1.00% 1.00% Tocopherol 1.00%
Dow Corning 200 Fluid 1.00% Soymilk Powder 5.00% Soybean Extract
using Ethanol/Water Mixture 5% 10% TOTAL 100% 100% 100% 100% 100%
100% 100%
[0091]
3TABLE 3 Soybean Essence formulations Soybean Essences 1 6 8 21 23
Soymilk 87.42% 89.04% 96.09% 96.05% 95.70% Phenoxyethanol 0.73%
Phenoxyethanol and Parabens 1.00% 1.00% 1.00% 1.00% Glycerin 2.50%
2.50% Cyclomethicone 2.00% Aluminum Starch Ocetyl Succinate 0.75%
Sucrose Cocoate 1.00% 1.00% PEG-6 Capric/Caprylic Triglycerides
3.00% 3.00% Disodium EDTA 0.10% 0.10% 0.05%
Polyacrylamide/Laureth-7C.sub.13-14 Isoparrafin 2.50% 2.75% 2.90%
2.90% 3.20% Ascorbic Acid 0.01% Butylated Hydroxytoluene 0.10%
0.01% 0.05% 0.05% Polysorbate 20 0.50% TOTAL 100% 100% 100% 100%
100%
[0092] While certain preferred embodiments of the present invention
have been described and specifically exemplified above, it is not
intended that the invention be limited to such embodiments. Various
modifications may be made thereto without departing from the scope
and spirit of the present invention, as set forth in the following
claims.
* * * * *