NZ615178B2 - Skin-beautifying agent - Google Patents
Skin-beautifying agent Download PDFInfo
- Publication number
- NZ615178B2 NZ615178B2 NZ615178A NZ61517812A NZ615178B2 NZ 615178 B2 NZ615178 B2 NZ 615178B2 NZ 615178 A NZ615178 A NZ 615178A NZ 61517812 A NZ61517812 A NZ 61517812A NZ 615178 B2 NZ615178 B2 NZ 615178B2
- Authority
- NZ
- New Zealand
- Prior art keywords
- skin
- whey protein
- beautifying
- protein hydrolysate
- group
- Prior art date
Links
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/40—Feeding-stuffs specially adapted for particular animals for carnivorous animals, e.g. cats or dogs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/40—Feeding-stuffs specially adapted for particular animals for carnivorous animals, e.g. cats or dogs
- A23K50/48—Moist feed
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/19—Dairy proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/54—Proteins
- A23V2250/542—Animal Protein
- A23V2250/5424—Dairy protein
- A23V2250/54252—Whey protein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/012—Hydrolysed proteins; Derivatives thereof from animals
- A61K38/018—Hydrolysed proteins; Derivatives thereof from animals from milk
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/007—Preparations for dry skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K4/00—Peptides having up to 20 amino acids in an undefined or only partially defined sequence; Derivatives thereof
- C07K4/10—Peptides having up to 20 amino acids in an undefined or only partially defined sequence; Derivatives thereof from plants
Abstract
Disclosed is the use of a whey protein hydrolysate in the manufacture of a medicament for treating dry skin.
Description
PTION
SKIN—BEAUTIFYING AGENT
TECHNICAL FIELD
The invention relates to a skin—beautifying agent that exhibits ent a
skin—beautifying effect, is effective for moisturizing, beautifying skin, preventing rough
skin, ting skin wrinkles, and preventing loss of elasticity in skin, has low
ness, and is excellent in stability and safety.
The invention also relates to a skin-beautifying cosmetic, a skin—beautifying
food or drink, a skin—beautifying nutrient composition, a skin-beautifying feed, and a
skin-beautifying drug ing the skin-beautifying agent.
OUND ART
Skin is an interface between a living body and the external environment, and
has a skin barrier function that prevents water loss from the body, and prevents
penetration of ically harmful substances, such as microorganisms, allergens and
the like from the external environment. Keratin ellular lipids, mainly ceramide,
sebum and the like in the stratum comeum act as these skin barrier function. The
stratum comeum must have a water content of 10 to 20% in order to achieve a normal
function and maintain a healthy condition. Therefore, softness and the elasticity of
skin are maintained by water retained in the stratum comeum due to the skin barrier
function. When the water content decreases in the stratum , skin loses its
softness and becomes hard, thereby cracks and the like may occur. So—called chapped
skin of which patterns are invisible or unclear shows a significant decrease in water
content in the stratum corneum. Rough skin not only has poor cosmetic appearance,
but also is apreliminary stage to cause skin disease, and rough skin has a pathological
cance. Further, by improving rough skin conditions, the surface of dry and flaky
skin can be smoothed, thereby leading to improvement of fine wrinkles. It has been
known that when the skin barrier function of the stratum comeum has deteriorated,
water loss from skin is significant in comparison with healthy condition, that is,
increased transepidermal water loss (TEWL) is observed. The TEWL is closely
related with the barrier on and the moisturizing function of the stratum comeum,
and is used as an index of the skin barrier function. Therefore, skin can be maintained
in a healthy condition i.e., beautiful condition by increasing the water t in skin, or
reducing the TEWL, or suppressing an increase in TEWL.
In recent years, studies on the mechanism of skin have been carried out, and it
has been confirmed that macroscopic causes of dry feeling of the skin and rough skin
are complexly involved in the effects of sunlight (ultraviolet rays), drying, oxidization,
and the like in on to effects due to decrement of the metabolism with aging. It
has been found that these effects caused by such factors significantly decrease the
amount of collagen fibers that are the main matrix component of the dermis. When a
mechanism to keep tension and elasticity of skin that is maintained by the en
fibers is destroyed by the effects of ultraviolet rays and the like, wrinkles or slacks of
the skin increase. Since en molecules can retain water, and thus maintain skin in
a moisturized condition,lskin becomes dry and rough when collagen is destroyed due to
external s. Therefore, wrinkles and slacks of skin can be prevented by promoting
biosynthesis of collagen that is one of the main components of the dermis layer, and
skin can be ined in a healthy condition, that is, beautifiil skin condition. In
recent years, deterioration in the skin condition of animals, particularly pets, due to
allergy and the like has become a major issue. The skin condition of animals can be
improved by rizing and caring the skin, healthy skin ion can be ined.
A milk protein hydrolysate has been used for various commercial products in
order to prevent food allergy caused by cow milk or dairy products. In particular, it
has been considered that a whey protein of cow milk functions as an allergen, differing
from a protein in mother’s milk. Therefore, it has been known that a whey protein has
been hydrolyzed using a protease in order to prevent this problem (see Patent
Documents 1 to 3). It has been also known that a whey n hydrolysate suppresses
the progress of atopic dermatitis (see Non-patent nt 1), and improves curing
wound after Caesarean section (see tent nt 2).
PRIOR ART DOCUMENT
PATENT DOCUMENT
Patent Document 1: JP-A-H02-002319
Patent Document 2: JP-A-H02-138991
Patent nt 3: JP-A-H04-112753
NON-PATENT DOCUMENT
Non-patent Document 1: Allergology International. 2006; 55: 185-189
Non-patent Document 2: British Journal of Nutrition 2010; Aug 9: 1-7
Y OF THE INVENTION
PROBLEMS TO BE SOLVED BY THE INVENTION
In some embodiments the present invention seeks to provide a skin-beautifying
agent that exhibits excellent beauty effects, such as excellent oisturizing,
skin-protecting, preventing or improving rough skin, preventing wrinkles, ting
loss of city in skin, or the like. In other embodiments the present invention seeks
to provide a skin-beautifying cosmetic, a skin-beautifying food or drink, a
skin-beautifying nutrient composition, a skin-beautifying feed, or a skin-beautifying
drug that includes a skin-beautifying agent that exhibits excellent beauty effects, such as
excellent skin-moisturizing, skin- protecting, preventing or improving rough skin,
preventing wrinkles, preventing loss of elasticity in skin, or the like.
MEANS FOR SOLVING THE PROBLEMS
The inventors of the invention ted extensive studies, and found that a
whey protein hydrolysate has a skin-beautifying effect.
Specifically, the invention includes the followings.
(1) A skin-beautifying agent including a whey protein ysate as an active
ingredient.
(2) The skin-beautifying agent according to (1), wherein the whey protein
hydrolysate has a hydrolysis rate of 25% or more.
(3) The eautifying agent according to (1) or (2), wherein the whey
n ysate has ing characteristics (A) to (D);
(A) the molecular weight thereof is 10 kDa or less and the main molecular
weight peak thereof is 200 Da to 3 kDa,
(B) the average peptide length (APL) thereof is 2 to 8,
(C) the free amino acid content hereof is 20% or less, and
(D) the antigenicity thereof is equal to or less than 1/100,000th of that of
β-lactoglobulin.
(4) The skin-beautifying agent according to any one of (1) to (3), wherein the
whey protein hydrolysate is obtained by yzing a whey protein using a
heat-resistant protease at pH 6 to 10 and 50 to 70oC while thermally denaturing the
whey protein, and inactivating the protease by heating.
(5) The skin-beautifying agent according to any one of (1) to (3), wherein the
whey protein hydrolysate is obtained by hydrolyzing a whey protein using a protease at
pH 6 to 10 and 20 to 55oC, heating the resultants to 50 to 70oC, further hydrolyzing the
unhydrolyzed whey protein using a heat-resistant protease at pH 6 to 10 and 50 to 70oC
while thermally denaturing, and inactivating the protease by g.
(6) A skin-beautifying cosmetic, a skin-beautifying food or drink, a
skin—beautifying nutrient ition, a skin-beautifying feed, or a skin—beautifying
drug including the skin-beautifying agent according to any one of (1) to (5).
(7) A method for moisturizing or protecting mammalian skin, preventing or
ing rough skin of mammalian, or preventing wrinkles or se in elasticity of
mammalian skin, comprising orally or parenterally administering a whey protein
hydrolysate.
(8) The method according to (7), wherein the whey protein hydrolysate has
following characteristics (A) to (D);
(A) the molecular weight thereof is 10 kDa or less and the main molecular
weight peak thereof is 200 Da to 3 kDa,
(B) the e peptide length (APL) f is 2 to 8,
(C) the free amino acid content thereof is 20% or less, and
(D) the antigenicity thereof is equal to or less than 1/10,000th of that of
B-lactoglobulin.
(9) The method according to (7), wherein the whey protein hydrolysate is
obtained by hydrolyzing a whey protein using a heat—resistant protease at pH 6 to 10 and
50 to 70°C While thermally ring the whey protein, and inactivating the protease
by heating.
(10) The method according to (7), wherein the whey protein hydrolysate has
following characteristics (A) to (E);
(A) the molecular weight thereof is- 10 kDa or less and the main molecular
weight peak thereof is 200 Da to 3 kDa,
(B) the average peptide length (APL) thereof is 2 to 8,
(C) the free amino acid content thereof is 20% or less,
(D) the antigenicity thereof is equal to or less than 00th of that of
B-lactoglobulin, and
(E) the average molecular weight thereof is 300 to 500 Da.
(11) The method according to (7), wherein the whey protein hydrolysate is
obtained by hydrolyzing a whey protein using a esistant protease at pH 6 to 10 and
50 to 70oC while lly denaturing the whey protein, and inactivating the protease
by heating, and then concentrating using an ultrafiltration (UF) membrane and/or a
microfiltlation (MF) membrane.
EFFECTS OF THE INVENTION
[0008a]
According to a first aspect the t invention provides a use of a whey
n hydrolysate for treating dry skin.
The skin-beautifying agent according to the invention exhibits a significant
skin-moisturizing effect and a skin collagen production-promoting effect, and may be
useful for moisturizing or protecting skin, preventing or improving rough skin,
preventing skin wrinkles, and preventing or treating loss of elasticity in skin. Since the
skin-beautifying agent according to the invention is produced using a whey protein as a
raw al, the eautifying agent can be easily and economically produced.
EMBODIMENTS FOR CARRYING OUT THE INVENTION
A whey protein hydrolysate that is included in a skin-beautifying agent of the
invention as an active ingredient may be obtained by the method disclosed in Patent
Document 3, for e. ing to the method disclosed in Patent Document 3, a
whey protein is adjusted to pH of 6 to 10 and a temperature of 50 to 70oC, and a
heat-resistant protease is added thereto to hydrolyze while thermally denaturing, and the
protease is inactivated by heating, y obtaining a whey protein hydrolysate. Note
that the yield of the whey protein hydrolysate can be further improved by preliminarily
hydrolyzing a whey protein using a protease at pH 6 to 10 and 20 to 55oC prior to the
above hydrolyzation by a heat-resistant protease, and then immediately hydrolyzing the
resultant under the above conditions without cooling the resultant. The whey n
hydrolysate prepared as described above may be concentrated using an iltration
(UF) membrane having a molecular weight cut-off of 1 to 20 kDa, preferably 2 to 10
kDa, and/or a microfiltration (MF) membrane having a molecular weight cut-off of 100
to 500 Da, preferably 150 to 300 Da. The bitterness of the whey protein hydrolysate
can be reduced while improving the transparency of the whey protein hydrolysate by
adjusting the average molecular weight of the whey protein hydrolysate to 300 to 500
Da utilizing the above membrane treatment.
[001 1]
The term “whey protein” used herein refers to whey prepared from milk of a
mammal, such as cow, goat, sheep, human or the like, an aggregate, a powder, or a
purified product thereof. The whey n is used in a state of an aqueous solution
when yzing the whey protein using a protease.
When preparing a whey protein hydrolysate according to the method disclosed
in Patent Document 3, the whey protein aqueous solution is adjusted to pH 6 to 10.
Note that the whey protein aqueous solution need not necessarily be adjusted the pH
thereof since a whey protein normally has a pH within the above range. When the
whey protein aqueous solution need be adjusted the pH, the whey protein s
solution is adjusted to pH 6 to 10 using a solution of an acid, such as hydrochloric acid,
citric acid, lactic acid or the like or an alkali, such as caustic soda, calcium hydroxide,
sodium phosphate or the like. The whey protein aqueous solution is heated to 50 to
70°C. It is preferable to add the heat-resistant protease before heating the whey protein
aqueous solution rather than at the above temperature from the viewpoint of the yield.
The optimum ature for a normal protease is 40°C or less, and the optimum
temperature for a esistant protease is 45°C or more. Any esistant protease
may be used without limitation as long as the protease is known as a heat~resistant
se having such an optimum temperature of 45°C or more. es of such a
heat—resistant-protease include papain, Protease S (trade name), Proleather (trade name),
ase (trade name), Alcalase (trade name), A (trade name), and the like. It
is preferable to use a heat—resistant protease that has a residual activity of about 10% or
more when heated at 80°C for 30 minutes. It is more effective to use a plurality of
ses in combination rather than to use only single protease. The reaction time is
preferably about 30 minutes to about 10 hours.
[00 1 3]
The protease is finally inactivated by heating the reaction solution. The
protease may be inactivated by heating the reaction solution at 100°C or more for 10
seconds or more. After centrifuging the reaction solution, the supernatant liquid is
collected, and dried to obtain a powdery product. Since a precipitate that occurs
during centrifugation is enic as compared with the atant liquid, it is
preferable to remove such a precipitate. Note that the reaction solution as it is may be
dried and sufficiently used. The average peptide length (APL) ofthe resulting whey
protein hydrolysate may be ined by a 2,4,6—trinitrobenzenesulfonate (TNB S)
method or the like. The molecular weight distribution of the whey protein hydrolysate
may be determined by high-performance size exclusion chromatography (HPSEC) or
the like. The free amino acid content in the whey protein hydrolysate may be
ined by extracting free amino acids using 75% ethanol or the like, and analyzing
the extracted free amino acids using an amino acid analyzer or the like. The hydrolysis
rate of the whey protein hydrolysate may be determined by an ortho—phthalaldehyde
(OPA) method, in which free amino groups are modified and measured, or the like.
The whey protein hydrolysate ofthe invention may be used ly as the
skin-beautifying agent, or may optionally be used as a preparation incorporated in a
powdered preparation, granules, a tablet, a capsule, a drinkable preparation, or the like
using a conventional method. A whey protein hydrolysate obtained using an
ultrafiltration (UF) membrane or a microfiltration (MF) membrane may also be used
directly as theskin—beautifying agent, or may also be used as the skin-beautifying agent
after directly drying. The whey protein hydrolysate may also be used as a ation
ed using a conventional method. The preparation may be added to a nutrient
preparation, a food or drink, such as , milk-based drink, wafer or the like, a
nt ition, a feed, or a drug.
A skin-beautifying. food or drink, a skin—beautifying nutrient composition, a
skin-beautifying feed, and a skin—beautifying drug of the invention may e only the
whey protein hydrolysate, or may further include raw materials, such as stabilizer,
saccharide, lipid, flavor, vitamin, mineral, flavonoid, polyphenol and the like, that are
normally ned in a food, drink, feed, or drug, for example. The whey protein
hydrolysate may be used in combination with additional ingredients that exhibits a
beauty effect, such as sphingomyelin, glucosylceramide, phospholipid and the like, that
exhibits a skin-moisturizing effect, and such as collagen, n C, iron and the like
that promote skin collagen production, for example. A raw material that is normally
contained in a food, drink, or thelike may be added to such a skin-beautifying food or
drink, skin-beautifying nutrient composition, skin-beautifying feed, or skin—beautifying
drug as another raw material.
The t of the whey n hydrolysate in the skin-beautifying food or
drink, the skin—beautifying nutrient ition, the skin-beautifying feed, and the
eautifying drug is not particularly limited, but is normally 0.001 to 10% (w/w),
and preferably 0.1 to 5% (w/w), based on the total mass of the food or drink, the
nutrient composition, the feed, and the drug so that an adult can take the whey protein
ysate in an amount of 2 mg/day or more, although the content of the whey protein
hydrolysate varies depending on the food or drink, the feed, and the drug. The
skin-beautifying cosmetic may be used as a normal cosmetic form such as emulsion,
cream, lotion, or pack. Such a cosmetic may be produced by a Conventional method,
and the whey protein hydrolysate may be appropriately added during the production
process. A ic may be produced using such a cosmetic as a raw material. The
content of the whey protein hydrolysate in the ic is not particularly limited, but is
normally 0.001 to 30% (w/w), and preferably 0.1 to 10% (w/w), based on the total mass
of the cosmetic in order to apply the whey protein hydrolysate in an amount of 2
mg/day or more per adult.
A skin-beautifying composition that can be orally or parenterally administered
may be prepared by adding an appropriate auxiliary agent to the active ingredient used
for the skin—beautifying agent of the invention, and preparing the composition in
optional form. When forming the composition, a diluent or vehicle such as a filler,
extender, binder, egrant, surfactant, lubricant or the like, that is normally added,
may be used. Various dosage form of a ceutical preparation may be selective,
for example, a capsule, tablet, granules, powder, liquid, suspension, emulsion,
suppository, ion, ointment and the like can be given. Examples ofthe vehicle
include sucrose, lactose, starch, crystalline cellulose, mannitol, light anhydrous silicic
acid, magnesium aluminate, synthetic aluminum silicate, magnesium
aluminometasilicate, calcium carbonate, sodium hydrogen carbonate, calcium hydrogen
phosphate, m carboxymethyl cellulose, and the like. These compounds may be
used either alone or in combination of two or more thereof.
[001 8]
The invention is further described below by way of examples, comparative
examples, and test examples. Note that the following examples are ed for
illustrative purposes only, and should not be construed as limiting the invention.
Example 1
[001 9]
Papain (50 U/g whey n) and Proleather (150 U/g whey protein;
manufactured by Amano Enzyme Inc.) were added to 1 liter of a 10% whey protein
s solution. After adjusting the pH of the mixture to 8, the whey n was
hydrolyzed and denatured at 55°C for 6 hours. The on on was then heated
at 100°C for 15 seconds or more to inactivate the proteases, and fuged. The
supernatant liquid was collected, and dried to obtain a whey protein hydrolysate
(Example product 1). The Whey protein hydrolysate had a molecular weight
distribution of 10 kDa or less, a main molecular weight peak of 1.3 kDa, an APL of 7.2,
and a free amino acid content of 18.9% (based on the total components). The whey
protein hydrolysate had antigenicity equal to or less than 1/10,000th of that of
B-lacto globulin red by inhibition ELISA). The hydrolysis rate was 28%, the
yield (i.e., the ratio (%) of the dry weight of the supernatant liquid to the dry weight of
the raw materials) was 80.3%, and the bitterness was 2. The whey protein ysate
thus obtained can be used directly as the skin-beautifying agent according to the
invention.
Example 2
Papain (50 U/g whey protein) and Proleather (150 U/g whey protein;
manufactured by Amano Enzyme Inc.) were added to 11 of a 10% whey protein
aqueous solution. After adjusting the pH of the mixture to 8, the whey protein was
hydrolyzed at 50°C for 3 hours. The solution was heated to 55°C, and kept the
ature for 3 hours to hydrolyze and denature the whey protein. The mixture was
then heated at 100°C for 15 seconds or more to inactivate the proteases. The reaction
solution was filtered using a UF membrane having a molecular weight cut—off of 10 kDa
(manufactured by STC) and an MP membrane having a molecular weight cut—off of 300
Da (manufactured by STC) to collect a concentrate on. The fraction was dried to
obtain a whey protein ysate le product 2). The whey protein hydrolysate
had a molecular weight distribution of 10 kDa or less, a main molecular weight peak of
500 Da, anAPL of 3 .0, and a fiee amino acid content of 15.2% (based on the total
ents). The whey n hydrolysate had antigenicity equal to or less than
1/10,000th of that of B~lactoglobulin (measured by inhibition ELISA). The hydrolysis
rate thereof was 32%, the yield was 65.4%, and the bitterness was 2. The whey
protein hydrolysate thus obtained can be used ly as the skin-beautifying agent
according to the invention.
Example 3
120 g of a whey protein was dissolved in 1800 ml of purified water, and the
solution was adjusted to pH 7.0 using a 1 M sodium hydroxide solution. The solution
was ized at 60°C for 10 minutes, and held at 45°C. After the addition of 20 g of
Amano A (manufactured byAmano Enzyme Inc.), the mixture was reacted for 2 hours.
After inactivating the protease by heating the mixture at 80°C for 10 s, the
mixture was -dried to obtain a whey protein hydrolysate (Example product 3).
The hydrolysis rate of the whey protein hydrolysate was 18%, and the yield was 80.6%.
The whey protein hydrolysate thus obtained can be used directly as the skin-beautifying
agent according to the invention.
Example 4
120 g of a whey protein was dissolved in 1800 m1 of d water, and the
on was adjusted to pH 7.0 using a l M sodium hydroxide solution. The solution
was sterilized at 60°C for 10 minutes, and held at 45°C. After the on of 20 g of
Amano A (manufactured by Amano Enzyme Inc.), the mixture was reacted for 8 hours.
After inactivating the protease by heating the mixture at 80°C for 10 minutes, the
mixture was freeze-dried to obtain a whey n hydrolysate (Example product 4).
The hydrolysis rate of the whey protein hydrolysate was 25%, and the yield‘was 80.6%.
The whey protein ysate thus obtained can be used directly as the skin—beautifying
agent according to the invention.
Comparative Example 1
200 g of casein was suspended in 2000 ml of purified water, and the solution
was ed to pH 8.0 using a 1 M sodium hydroxide solution and dissolved
completely. The solution was sterilized at 80°C for 10 minutes, and held at 50°C.
After the addition of 20 g of Pancreatin F (manufactured byAmano Enzyme Inc.) and
g ofAmano A (manufactured byAmano Enzyme Inc.), the mixture was reacted for
hours. After inactivating the proteases by heating the mixture at 80°C for 10
minutes, the mixture was freeze-dried to obtain a casein hydrolysate (Comparative
example product 1). The hydrolysis rate of the casein ysate was 27%, and the
yield was 77.8%.
Comparative Example 2
200 g of casein was suspended in 2000 ml of purified water, and the solution
was adjusted to pH 8.0 using a 1 M sodium hydroxide on and ved
completely. The solution was sterilized at 80°C for 10 minutes, and held at 40°C.
After the addition of 15 g of Pancreatin F (manufactured byAmano Enzyme Inc.), the
mixture was reacted for 5 hours. After inactivating the se by heating the mixture
at 80°C for 10 minutes, the e was freeze-dried to obtain a casein hydrolysate
(Comparative example product 2). The hydrolysis rate of the resulting casein
hydrolysate was 20%, and the yield was 79.1%.
Test Example 1
Animal experiment
A skin—moisturizing effect was evaluated using whey n hydrolysates of
Example products 1, 3, and 4 and casein hydrolysates of Comparative example products
1 and 2. l3-week—old ss mice (Hos: HR—l) were used for the experiment. The
mice were divided into following eight groups (8 mice/group); a group to which
physiological saline was stered (Group A), a group to which whey protein
hydrolysate of Example product 1 was administered in an amount of 2 mg/kg
weight(Group B), a group to which whey protein ysate of Example product 1 was
administered in an amount of 5 mg/kg weight(Group C), a group to which whey protein
hydrolysate of Example product 1 was administered in an amount of 10 mg/kg
weight(Group D), a group to which whey protein hydrolysate of Example product 3 was
administered in an amount of 10 mg/kg weight(Group E), a group to which whey
protein hydrolysate of Example product 4 was administered in an amount of 10 mg/kg
weight(Group F), a group to which casein hydrolysate of ative example product
1 was stered in an amount of 10 mg/kg weight(Group G), and a group to which
casein hydrolysate of Comparative example product 2 was administered in an amount of
mg/kg weight(Group H). Each mouse was orally administered once a day using a
sonde, and kept for 3 weeks. Example products 1, 3, and 4 and ComparatiVe example
products 1 and 2 were suspended in physiological saline, and orally administered to the
Groups B to H. The moisture content in the tail of each mouse and the transepidermal
water loss from the tail of each mouse were measured at the beginning and the end of
the test, and the values (the moisture content and the transepidermal water loss) at the
end of the test were calculated as a ratio (increase ratio) with respect to the values at the
ing of the test which were assumed 100. The moisture content in the skin and
the transepidermal water loss were respectively measured using a Corneometer and a
Tewameter (manufactured by Courage & Khazaka). The results are shown in Table 1.
Moisture content Transepidermai water loss
increase ratio (%) Increase ratio (%)
As shown in Table 1, while Group A showed that the moisture content
decreased after 3 weeks of administration, Group B showed that the re content
increased about 1.25 times after 3 weeks of administration, and Groups C and D showed
that the moisture content increased up to about 1.5 times after 3 weeks of administration.
Groups E and F showed that the moisture content increased up to about 1.2 times after 3
weeks of administration. In contrast, in the cases of the groups to which casein
ysate of Comparative example t 1 or 2 was administered, both the moisture
content and pidermal water loss did not change between the beginning and end of
the test. It was confirmed from these results that the whey protein hydrolysate
according to the invention exhibited a oisturizing effect. It was also confirmed
that the skin-moisturizing effect was achieved when the whey protein hydrolysate was
administered in an amount of at least 2 mg/kg weight.
Test e 2
Animal experiment
A collagen tion-promoting effect was evaluated using whey protein
hydrolysate of Example product 2 and casein hydrolysates of C0mparative example
products 1 and 2. 7-week—old Wistar male rats were d into following six groups
(6 mice/group); a group to which physiological saline was administered (Group A), a
group to which whey protein ysate of Example product 2 was. administered in an
amount of 2 mg/kg weight (Group B), a group to which whey protein hydrolysate of
Example product 2 was administered in an amount of 5 mg/kg weight (Group C), a
group to which whey protein hydrolysate of Example product 2 was administered in an
amount of 10 mg/kg weight (Group D), a group to which casein hydrolysate of
Comparative example product 1 was administered in an amount of 10 mg/kg weight
(Group E), and a group to which casein hydrolysate of ative example product 2
was administered in an amount of 10 mg/kg weight (Group F). Each rat was
administered once a day using a sonde, and kept for 10 weeks. Example product 2 and
Comparative e products 1 and 2 were suspended in physiological saline, and
orally administered to Groups B to F. The amount of collagen in the skin was
determined by treating the derma of each rat in accordance with the method ofNimni et
al. (see Arch. Biochem. Biophys., p. 292, 1967), and measuring the hydroxyproline
content in the soluble fraction. Hydroxyproline is a l amino acid that is
ned only in collagen, and accounts for about 10% of the amino acids that form
collagen. Therefore, the amount of collagen can be estimated by measuring the
hydroxyproline content (see Asano Ryuji et al., Biolndustry, p. 12, 2001). The results
are shown in Table 2.
TABLE 2
Hydroxyproline content (pg/ml)
GroupA 0.3 i 0.1
The values shown in Table 2 indicate “mean i standard ion” (n=6).
The symbol “*” indicates that there was a icant ence as compared with
Group A (control group) 5).
As shown in Table 2, the amount of hydroxyproline in the soluble fraction after
weeks of administration was significantly larger in Groups B, C, and D as compared
with Group A. Groups E and F did not show a difference in the amount of
hydroxyproline in the soluble fraction after 10 weeks of administration from Group A.
It was confirmed from these results that the whey protein ysate according to the
invention exhibited a skin collagen production-promoting effect. It was also
confirmed that the skin collagen production-promoting effect was achieved when the
whey protein hydrolysate was administered in an amount of at least 2 mg/kg weight.
Example 5
[003 1]
Preparation of skin—beautifying cosmetic (lotion)
A lotion—type skin—beautifying cosmetic (Example product 5) was prepared by
mixing the raw materials in the composition shown in Table 3.
TABLE 3
Sorbitol
Sodium DL-pyrrolidone carboxylate
Carboxymethyl cellulose 0.3
Parabene 0.1
Sterilized ion—exchanged water Balance (total: 100)
Preparation of skin—beautifying cosmetic (cream)
A cream—type skin-beautifying cosmetic (Example product 6) was prepared by
mixing the raw materials in the composition shown in Table 4.
TABLE 4
Glycerol monostearate (self—emulsifiable)
ed n
Liquid paraffin
Jojoba oil
Parabene
Example product 1 0.5
Flavor Proper quantity
Sterilized ion—exchanged water Balance (total: 100)
[003 5]
Test Example 3
A clinical test was performed using Example product 5 and Example product 6.
A comparative lotion and a comparative cream were produced in the same manner as
e product 5 or Example t 6, except that the whey protein hydrolysate was
not added. Twenty adult females having face with dry skin, slacks and fine wrinkles of
the skin were divided at random into two groups each having 10 subjects (Groups A and
B), and twenty adult females having hands and fingers with rough skin were divided at
random into two groups each having 10 subjects (Groups C and D). 2 g of Example
product 5, 2 g of the comparative lotion, 2 g of Example product 6, and 2 g ofthe
comparative cream were applied to the faces of Group A, the faces of Group B, the
hands and fingers of Group C, and the hands and fingers of Group D, tively, in
the same way as usual twice a day for 10 days. The results are shown in Table 5.
TABLE 5
Wrinkles Slacks
++ ;A significant effect was observed after 10 days of application
+ ct was observed after 10 days of application
:t ; Any effect was not observed after 10 days of ation (same as ten days
before)
As shown in Table 5, it was confirmed that Example product 5 significantly
improved skin dryfeeling, rough skin and the like, and exhibited a skin—beautifying
effect as compared with the comparative lotion. It was also confirmed that Example
product 6 cantly improved skin dry feeling, rough skin, and suppressed
spontaneous exacerbation, such as rough skin or the like as compared with the
comparative cream.
Example 7
Preparation of skin-beautifying tablet
The raw materials were mixed in the composition shown in Table 6. 1 g of
the ing mixture was formed and tableted by a conventional method to obtain a
skin-beautifying tablet according to the invention. 1 g ofthe skin-beautifying tablet
contained whey protein ysate ofExample product 1 in an amount of 50 mg.
TABLE 6
Hydrous crystalline glucose 88.5 (wt%)
Mineral mixture 5.0
Example product 1 . 5.0
Sugar ester 1.0
Example 8
Preparation of skin-beautifying liquid nt composition
50 g of whey the protein hydrolysate ofExample t 2 was dissolved in
4950 g of deionized water. The solution was heated to 50°C, and mixed with stirring
at 6000 rpm for 30 minutes using a TK-homomixer (“TK ROBOMICS” manufactured
by PRIMIX Corporation) to obtain a whey protein hydrolysate solution which contained
the whey protein hydrolysate of Example product 2 in an amount 50 g/5 kg. 5.0 kg of
casein, 5.0 kg of soybean protein, 1.0 kg of fish oil, 3.0 kg of perilla oil, 17.0 kg of
dextrin, 6.0 kg of mineral mixture, 1.95 kg ofvitamin mixture, 2.0 kg of emulsifier, 4.0
kg of stabilizer, and 0.05 kg of flavor were added to 5.0 kg of the whey protein
hydrolysate solution. The resulting mixture was d in a 200 m1 retort pouch.
The mixture was then sterilized at 121°C for 20 minutes using a retort sterilizer -1
pressure vessel, CRTGN” manufactured by Hisaka Works, Ltd.) to obtain 50 kg
of a skin-beautifying liquid nt composition according to the invention. The
skin-beautifying liquid nutrient composition contained the whey protein hydrolysate of
Example product 2 in an amount of 100 mg/100 g.
Example 9
ation of skin-beautifying drink
300 g of a skim milk powder was dissolved in 409 g of zed water, and 1
g of the whey n hydrolysate of Example prOduct 1 was dissolved in the solution.
The solution was heated to 50°C, and mixed with stirring at 9500 rpm for 30 minutes
using an ultra-disperser (“ULTRA-TURRAX T-25” manufactured by IKA Japan).
After the addition of 100 g of maltitol, 2 g of acidifier, 20 g of reduced starch syrup, 2 g
of flavor, and 166 g of deionized water, the resulting mixture was charged in a 100 m1
glass . After sterilizing the mixture at 95°C for 15 minutes, the bottle was closely
sealed, thereby 10 bottles (100 ml each) of a skin—beautifying drink ofthe invention
were prepared. The skin-beautifying drink ned the whey protein hydrolysate of
Example product 1 in an amount of 100 mg/100 m1.
Example 10
Preparation of skin-beautifying dog food
2 kg ofthe Whey protein hydrolysate of Example product 2 was dissolved in 98
kg of deionized water. The solution was heated to 50°C, and mixed for stirring at 3600
rpm for 40 minutes using a TK—homomixer (“MARK II 160” manufactured by PRIMIX
Corporation) to obtain a whey protein hydrolysate solution which contained the whey
protein hydrolysate of Example product 2 in an amount of 2 g/100 g. 12 kg of soybean
meal, 14 kg ofpowdered skim milk, 4 kg of soybean oil, 2 kg of corn oil, 23.2 kg of
palm oil, 14 kg of corn starch, 9 kg of flour, 2 kg of bran, 5 kg of vitamin mixture, 2.8
kg of cellulose and 2 kg of l mixture were added to 10 kg of the whey protein
hydrolysate solution. The mixture was sterilized at 120°C for 4 minutes to obtain 100
kg of a skin-beautifying dog food ing to the invention. The eautifying
dog food contained the whey protein hydrolysate of Example product 2 in an amount of
200 mg/100 g.
hout this specification and the claims which follow, unless the context
requires otherwise, the word "comprise", and variations such as "comprises" or
ising", will be understood to imply the inclusion of a stated integer or step or
group of integers or steps but not the exclusion of any other integer or step or group of
integers or steps.
The reference in this specification to any prior publication (or information
derived from it), or to any matter which is known, is not, and should not be taken as an
acknowledgment or ion or any form of suggestion that that prior publication (or
information derived from it) or known matter forms part of the common general
knowledge in the field of endeavour to which this specification relates.
Claims (3)
1. Use of a whey protein hydrolysate in the manufacture of a medicament for ng dry skin.
2. The use according to claim 1, n the whey protein hydrolysate has a hydrolysis rate of 25% or more.
3. The use according to claim 1 or claim 2, wherein the whey protein hydrolysate has the following characteristics (A) to (D); (A) the molecular weight thereof is 10 kDa or less and the main molecular weight peak thereof is 200 Da to 3 kDa, (B) the average peptide length (APL) thereof 2 to 8, (C) the free amino acid content thereof 20% or less, and (D) the antigenicity thereof is equal to or less than
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2011-053276 | 2011-03-10 | ||
JP2011053276A JP2012188384A (en) | 2011-03-10 | 2011-03-10 | Skin-beautifying agent |
PCT/JP2012/055811 WO2012121286A1 (en) | 2011-03-10 | 2012-03-07 | Skin-beautifying agent |
Publications (2)
Publication Number | Publication Date |
---|---|
NZ615178A NZ615178A (en) | 2015-08-28 |
NZ615178B2 true NZ615178B2 (en) | 2015-12-01 |
Family
ID=
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