AU2019236633A1 - Cosmetic composition comprising Mangifera indica irwin seed extract - Google Patents
Cosmetic composition comprising Mangifera indica irwin seed extract Download PDFInfo
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- AU2019236633A1 AU2019236633A1 AU2019236633A AU2019236633A AU2019236633A1 AU 2019236633 A1 AU2019236633 A1 AU 2019236633A1 AU 2019236633 A AU2019236633 A AU 2019236633A AU 2019236633 A AU2019236633 A AU 2019236633A AU 2019236633 A1 AU2019236633 A1 AU 2019236633A1
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- irwin
- cosmetic composition
- extract
- mangifera indica
- skin
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- 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
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/52—Stabilizers
- A61K2800/522—Antioxidants; Radical scavengers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The present invention relates to a cosmetic composition or a
composition of skin external preparation comprising Mangifera indica irwin seed
extract as an active ingredient, and more particularly, the Mangifera indica irwin
5 seed extract has antimicrobial activity, anti-inflammatory activity and antioxidant
activity against skin flora and therefore it can be utilized as a cosmetic
composition or a composition of skin external preparation for improving anti
aging or skin inflammation and it effectively improves the skin inflammation and
prevents the aging.
10
24
1/3
DRAWINGS
FIG. 1
100.00 m.vit-c
iTMD
NTME
S600
00
S2000O
5 10 2S 5o 100 Soo 1000
Concentration (pg/mi)
FIG. 2
5 E Coi 5epidrmkh& Saureus
25
Description
1/3
FIG. 1 100.00 m.vit-c
iTMD
S600
00
S2000O
5 10 2S 5o 100 Soo 1000 Concentration (pg/mi)
FIG. 2
E Coi 5epidrmkh& Saureus
This application claims priority to and the benefit of Korean Patent
Application No. 10-2019-0105979 filed in the Korean Intellectual Property Office
on 28, 08, 2019, the entire contents of which are incorporated herein by
reference.
1. Field of the Disclosure
The present disclosure relates to a cosmetic composition comprising
Mangifera indica irwin seed extract as an active ingredient.
2. Description of the Related Art
Recently, many people have continued to be exposed to various
environments and their interest in health has increased. As the consumption of
natural antioxidants using natural products is higher than products containing
synthetic antioxidants and the interest in natural raw material for cosmetics
increases, the development of cosmetics using natural products increases and
as a new material which can effectively settle the oxidative stress and skin
aging, materials of naturalism and in various biotechnology are attracting
attention.
Oxidation which occurs mainly in our body, generates large amounts of
reactive oxygen species due to metabolic processes, and these reactive oxygen species are mostly degraded in the body. However, when the balance of defense system is broken, oxygen free radicals are not decomposed but accumulated in the body and cause cellular tissue damage due to oxidative stress, which can cause various diseases. Reactive oxygen species are necessary for cell growth, but if the amount of production increases due to environmental pollution, exposure to chemicals, stress, obesity, or the function of the elimination system is weakened, it causes oxidative damage and causes various diseases. In addition, the sebum secreted from the skin due to external influences gathers, thereby increasing the activity of the skin flora which causes skin inflammation and accelerating inflammation and aging of the skin.
Inflammatory response is a biological defense mechanism in which cells
in the body protect the human body from skin damage caused by stimulation
such as physical shock or bacterial infection, etc. The regulation of the
inflammatory response in the body is involved in various immune cells for
abnormal stimulation, of which macrophages are known to be involved in
maintaining homeostasis by participating in various host responses as well as
innate immunity. Particularly, enzymes related to the biosynthesis of nitric oxide
synthase (NOS) and prostaglandin (PG) are known to play an important role in
mediating the inflammatory response. Therefore, NOS, an enzyme producing
nitric oxide(NO) from L-Arginine and cyclooxygenase(COX), an enzyme
involved in synthesizing prostagladins from arachidonic acid, have been major
goal in blocking inflammation.
Mango (Mangifera indica) is classified as the Anacardiaceae Mangifera
and its scientific name is Mangifera indica. Mango is an evergreen tree of the
Dicotyledoneae, Spondias dulcis and is about 15-18 m high and the leaf looks
like a spear and is about 30 cm long. As one of the top 30 crops in the world, it
is a major crop grown in tropical and subtropical regions. Currently, there is a
high interest in growing tropical fruit trees, a new high-income crop using vinyl
greenhouses in Jeju Island as measures for dealing with the climate change
due to global warming, and thus mangoes are cultivated.
There are more than 2,000 varieties of mangoes, and research has
reported that there are big differences in color, fragrance, and fiber of fruit
depending on the varieties. There are various polyphenol compounds in
mangoes. At present, many bioactive researches on mango skin and leaves
have proven, but studies on the bioactivity using mango seeds which are being
discarded in large quantities, are insufficient.
[Prior Art Documents]
[Patent literature]
Korea Patent No. 10-1872458 (Registered on June 22, 2018)
In order to solve the above problems, the present invention provides a
cosmetic composition or a composition of skin external preparation comprising
seed extract of Mangifera indica irwin which is natural raw material as an active
ingredient.
The cosmetic composition according to the present invention may
comprise Mangifera indica irwin seed extract as an active ingredient.
The composition of skin external preparation according to the present invention may comprise Mangifera indica irwin seed extract as an active ingredient.
In addition, a composition for inhibiting growth of skin flora according to
the present invention may comprise Mangifera indica irwin seed extract as an
active ingredient.
FIG. 1 is a graph measuring the electron donating ability according to
the concentration of Mangifera indica irwin seed extract of an experimental
example of the present invention.
FIG. 2 is an image showing the antimicrobial activity of Mangifera indica
irwin seed extract according to an experimental example of the present
invention.
FIG. 3 is a graph measuring cell viability according to an experimental
example of the present invention.
FIG. 4 is a graph measuring the nitric oxide (NO) production rate
according to an experimental example of the present invention.
FIG. 5 is a result confirming the expression of proinflammatory factors
according to an experimental example of the present invention.
Hereinafter, the present invention will be described in detail.
The present inventors have found various physiologically active
characteristics such as antioxidant and anti-inflammatory activity in the
Mangifera indica irwin seed extract which is a natural material and confirmed
the possibility of Mangifera indica irwin seed extract as a cosmetic natural
material, thereby completing the present invention.
As used herein, the term "extract" refers to a substance obtained by
extracting a component of a natural product regardless of an extraction method,
an extraction solvent, an extracted component or an extract form and it may
include all materials that can be obtained by other processing method after
extracting a substance obtained by extracting a component of a natural product.
As used herein, the term "cosmetic" may include a functional cosmetic
having an effect of preventing or improving skin aging, etc., and the functional
cosmetic has a professional functionality with the emphasis of the physiological
activity efficacy or effects unlike general cosmetics. As defined in the Cosmetic
Act, it refers to cosmetics that emphasize specific efficacy and effects, such as
skin whitening and skin wrinkle improvement.
The present invention provides a cosmetic composition comprising
Mangifera indica irwin seed extract as an active ingredient
In the cosmetic composition according to the present invention, the
Mangifera indica irwin seed is a seed portion which the pulp is removed from
the Mangifera indica irwin, and preferably may be a seed of Irwin which grows
naturally in Jeju Island. The apple mango native to Jeju Island is a variety of
apple mangoes produced in subtropics and has high adaptability to domestic
soil. Compared to the products obtained in foreign countries with a lot of fiber, thick skin, and large seeds, the improved mango of Jeju Island has a thin skin and small seeds, high sugar content and excellent fragrance. The apple mango seed may be obtained from apple mango collected or cultivated in nature, or may be purchased commercially available.
In the cosmetic composition according to the present invention, the
Mangifera indica irwin seed extract may be extracted with water, C1 to C4
alcohol or a mixed solvent thereof, and preferably it may be hot water extract or
60 to 80 weight% ethanol aqueous solution extract, and more preferably it may
be an extract of 70 weight% ethanol aqueous solution.
The Mangifera indica irwin seed extract may be extracted according to
method commonly used in the art, for example, hot water extraction, ultrasonic
extraction, filtration, reflux extraction and the like, and it can be performed alone
or at least two methods in combination.
In the cosmetic composition according to the present invention, the
Mangifera indica irwin extract may have an antimicrobial activity. More
specifically, the Mangifera indica irwin extract may have an antimicrobial activity
against at least one skin flora selected from the group consisting of Escherichia
coli (E. coli), Staphylococcus aureus (S. aureus) and Staphylococcus
epidermidis (S. epidermidis).
As used herein, the term "antimicrobial" or "antimicrobial activity" is a
property that resists microorganisms such as bacteria and fungi, and more
specifically, it means a feature that antibiotics, etc. inhibit the growth or
proliferation of the microorganisms.
As used herein, "skin flora" is a microorganism that has maintained a
close relationship with a specific tissue of a living body such as skin, and also
protects the skin from harmful bacteria and has a beneficial effect, but when it is
excessively proliferated or generates harmful substances, for people with low
resistance, this can be an opportunistic bacterium and cause skin inflammation
diseases and the like.
According to one experimental example of the present invention, the
antimicrobial activity was increased according to the concentration of ethanol
extract of Mangifera indica irwin seed, and all the strains of E. coli, S. aureus
and S. epidermidis showed relatively high antimicrobial activity. And in particular,
E. coli showed the highest antimicrobial activity.
In the cosmetic composition according to the present invention, the
Mangifera indica irwin seed extract may have anti-inflammatory activity. More
particularly, the Mangifera indica irwin seed extract may reduce the production
of nitric oxide (NO) or inhibit the expression of cyclooxygenase-2 (COX-2).
In the present specification, "anti-inflammatory" means the inhibition of
inflammation, the inflammation is an immune response of the human body in
response to a wound or disease, and oxidative stress such as ultraviolet rays,
oxygen free radicals and free radicals activates inflammatory factors and
causes various diseases and the skin aging.
According to one experimental example of the present invention, the NO
production of the cells was rapidly increased bylipopolysaccharide (LPS), but
when the Mangifera indica irwin seed hot water extract or Mangifera indica irwin seed ethanol extract was treated, the NO production rate was found to be reduced in a concentration-dependent manner.
In addition, regarding the measurement of the expression level of the
COX-2 protein, when the extract was treated, the expression level of COX-2
protein increased by LPS treatment was found to be decreased in a
concentration-dependent manner.
In the cosmetic composition according to the invention, the Mangifera
indica irwin seed extract may have an antioxidant activity.
As used herein, the term "antioxidant" refers to the action of inhibiting
oxidation and oxidative stress in vivo causes cell damage and pathological
diseases, and may also be a direct cause of aging.
According to one experimental example of the present invention, the
Mangifera indica irwin seed extract was found to have a high content of an
antioxidant polyphenol, and when free radical scavenging activity was
measured, it exhibited to increase in concentration-dependent manner,
confirming that it had an excellent antioxidant effect.
Since the cosmetic composition according to the present invention has
antimicrobial activity, anti-inflammatory activity and antioxidant activity as
described above, it can be utilized as a cosmetic composition for anti-aging or
improving the skin inflammation.
As used herein, the term "anti-aging" refers to inhibiting the aging or
reversing the aging, the skin constituents change or decrease as the aging progresses, subject to oxidative stress caused by free radicals and reactive harmful oxygen and skin wrinkles increase or elasticity decreases.
Through the experimental example of the present invention, the
antimicrobial effect on the skin flora causing skin inflammatory diseases in the
Mangifera indica irwin seed extract and the effect on the improvement of the
skin inflammation and the prevention of skin aging were confirmed and thus a
cosmetic composition comprising the Mangifera indica irwin seed extract as an
active ingredient can be utilized for anti-aging or skin inflammation improvement.
In the cosmetic composition according to the present invention, the
Mangifera indica irwin seed extract may be included in 0.001 to 50 parts by
weight based on 100 parts by weight of the total cosmetic, but it is not limited
thereto.
The cosmetic composition according to the present invention may
further comprise one or more of the ingredients listed in the list of cosmetic raw
materials registered with the Food and Drug Administration and ingredients
listed in ICID (International Cosmetic Ingredient Dictionary). For example, the
cosmetic composition may include at least one of an organic solvent, a
dissolving agent, a thickening agent, a gelling agent, a softening agent, an
antioxidant, a suspending agent, a stabilizer, a fragrance agent, a surfactant, an
emulsifier, a filler, a metal ion sequestrant, a preservative, a vitamin, a blocking
agent, a wetting agent, dyes and pigments, hydrophilic or lipophilic active
agents or auxiliary agents commonly used in the cosmetic field, such as any
other ingredient conventionally used in cosmetics.
The vitamin may include a water-soluble or oil-soluble vitamin, the
water-soluble vitamin can be formulated in cosmetics, preferably vitamin B1,
vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12,
pantothenic acid, nicotinic acid, nicotinamide, folic acid, vitamin C, vitamin H,
and the like, and more preferably may include vitamin C to help collagen
synthesis. In addition, salts thereof (thiamine hydrochloride, sodium ascorbate
salt, etc.) or derivatives thereof (ascorbic acid-2-sodium phosphate salt,
ascorbic acid-2-magnesium phosphate salt, etc.) may also be included and
water-soluble vitamin can be obtained by a conventional method such as
microbial transformation method, purification method from culture of
microorganisms, enzyme method or chemical synthesis method.
The cosmetic composition according to the present invention may be
prepared in any formulation conventionally prepared in the technical field to
which the present invention pertains. For example, it may be formulated as
toners, emulsions, lotions, creams, pastes, gels, solutions, suspensions, oils,
waxes, packs, powders, foundations, sprays, surfactant-containing cleansing,
and the like, but it is not limited thereto. More specifically, it may be prepared in
the formulation of softening lotion, nourishing lotion, astringent lotion, nourishing
cream, massage cream, milk lotion, powder, essence, eye cream, sun lotion,
sun cream, makeup primer, makeup base, BB cream, powder foundation,
emulsion foundation cleansing cream, cleansing foam, cleansing water, soap,
pack, stick-form product, balm type product, spray or powder.
When the cosmetic composition according to the present invention is a cream or gel formulation, it may further include animal oil, vegetable oil, wax, paraffin, starch, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide and the like, as the carrier component
When the cosmetic composition is a solution or emulsion formulation, it
may further include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate,
benzyl alcohol, propylene glycol, glycerol aliphatic ester, fatty acid ester of
polyethylene glycol or sorbitan, as a solvent, a solvating agent or an emulsifying
agent.
When the cosmetic composition is a suspension formulation, it may
further include liquid diluents such as ethanol or propylene glycol, suspending
agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters,
and polyoxyethylene sorbitan esters, microcrystalline cellulose, aluminum
metahydroxide, bentonite, agar or tragacanth, as the carrier component.
When the cosmetic composition is a powder or spray formulation, it may
further include lactose, talc, silica, aluminum hydroxide, calcium silicate or
polyamide powder, and in particular, in the case of spray formulation, it may
additionally include chlorofluorohydrocarbon, a propellant such as
propane/butane or dimethyl ether, as the carrier component.
When the cosmetic composition is a surfactant-containing cleansing
formulation, it may further include an aliphatic alcohol sulfate, an aliphatic
alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an
imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide,
ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glycerides, fatty
acid diethanolamide, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters, as the carrier component.
The cosmetic composition according to the present invention may be
coated and used alone or in duplicate, or may be coated and used in
combination with other cosmetic compositions other than the present invention.
In addition, the cosmetic composition according to the present invention may be
used according to a conventional method of use, and the number of times of
use may vary depending on the skin condition or preference of the user.
The present invention provides a composition of skin external
preparation for anti-aging or improving skin inflammation comprising Mangifera
indica irwin seed extract as an active ingredient.
Because the Mangifera indica irwin seed extract has antimicrobial
activity, anti-inflammatory activity and antioxidant activity against skin flora that
causes skin inflammatory diseases as described above, and thus it can be used
as a composition of skin external preparation for anti-aging or improving skin
inflammation.
As used herein, "skin external preparation" is a formulation that can be
directly applied to the outside of the skin, and may include a cosmetic
composition, and the formulation may include an ointment, a plaster, a patch, a
liniment, a pasta or a cataplasma and the like, but it is not limited thereto.
In addition, the present invention provides a composition for inhibiting
growth of skin flora comprising Mangifera indica irwin seed extract as an active
ingredient.
Hereinafter, examples will be described in detail to help understand the
present invention. However, the following examples are merely to illustrate the
content of the present invention, but the scope of the present invention is not
limited thereto. The embodiments of the present invention are provided to more
completely explain the present invention to those skilled in the art.
<Example 1> Preparation of Mangifera indica irwin seed extract
Mangifera indica irwin seed used in an experimental example according
to the present invention was used after removing the pulp of the Mangifera
indica irwin native to Jeju Island, drying at room temperature and storing at 4 °C.
The sample was extracted at 100 °C for 9 hours by adding water of about 10
times the weight of the sample (Heating Mantle, MS-DM608, MTOP6, Korea),
and extracted by shaking at room temperature for 24 hours with 70% ethanol.
The extraction was repeated three times in the same manner. Water and
ethanol extracts were filtered using vacuum filter paper (Whatman) NO.4 and
NO.2 (vacuum pump, DOA-P704-AC, GAST, USA) In turn, followed by
concentrating by a rotary vacuum evaporator (N-100, EYELA, Tokyo Japan)
(40 °C, speed 4-5), and used as a sample of the following experiment while
freeze-dried and stored in the freezer.
The following experimental results are expressed as Mean±SD
(Standard deviation), and NOVA of the experimental data obtained from the
control group and each sample and if the experimental data analyzed have
significance, the Turkey range test was performed as post-hoc test to verify the significance at the 95% level.
<Experimental Example 1> Confirmation of total polyphennols
content
The total polyphenol content of the Mangifera indica irwin seed extract
prepared according to an example of the present invention was measured by
the modified Folin-Denis method (Folin-Denis, 1912). 2 ml of the Mangifera
indica irwin seed extract diluted 10-fold in distilled water and 2 ml of 50% Folin
ciocalteu reagent were mixed and allowed to stand at room temperature for 3
minutes. 10% CaCO22 ml was added thereto, mixed, and the mixture was left at
room temperature for 1 hour and then the absorbance at 700 nm was measured.
The following Table 1 shows the total polyphenol content of the
Mangifera indica irwin seed extract according to the extraction method.
[Table 1]
Mangifera indica irwin seed Mangifera indica irwin seed hot water extract (TMD) ethanol extract (TME) (Totalpolyphennols 214.65 ±0.67 380.09 ±0.75 (mg/g dry)
Referring to the Table 1, as a result of measuring the polyphenol content
contained per each extract powder, the Mangifera indica irwin seed hot water
extract (TMD) is 214.65±0.67 mg/g, the Mangifera indica irwin seed ethanol
(70% ethanol) extract (TME) showed a high content of 380.09±0.75 mg/g.
When compared to the standard material tannic acid, it was confirmed that it
has high polyphenol content.
<Experimental Example 2> Confirmation of electron donation
ability
Radical scavenging activity against 1,1-diphenyl-2-picryl hydrazyl
(hereinafter referred to as DPPH) was measured by the modified Blois (1958)
method. 3 ml of 60 pM DPPH (Sigma, USA) was added to 0.5 ml of the diluted
sample at concentrations (5, 10, 25, 50, 100, 500, 1,000pg/ml), vortexed for 5
minutes and left for 15 minutes and the absorbance was measured at 517 nm.
FIG. 1 is a graph measuring the electron donating ability according to
the concentration of the Mangifera indica irwin seed extract according to an
experimental example of the present invention. Electron donating ability is used
as a measure of the reactive oxygen action by the active radicals in the human
body and is shown as the graph by converting the scavenging rate in terms of
percentage. Here, Vit-C (vitamin C) is a control, TMD means the Mangifera
indica irwin seed hot water extract, TME means the Mangifera indica irwin seed
ethanol extract and the value of the graph represents the average of the three
measurements.
Referring to FIG. 1, the electron donating ability of each sample at an
initial concentration of 5 pg/ml of hot water extract (TMD) was 7.45±0.62 % and
that of ethanol extract (TME) was 16.40±0.88 %, and the effect was 90% and
more at concentration of 100 pg/ml. Compared with the control group Vit-C,
each extract showed a significant tendency and increased in all extracts in a
concentration-dependent manner.
<Experimental Example 3> Confirmation of antibacterial activity
The disclosed strains used in the antibacterial activity test according to
the present invention are skin flora such as Escherichia coli (hereinafter, E.coli,
KCTC 1039), Staphylococcus aureus (hereinafter, S. aureus, KCTC 1621) and
Staphylococcus epidermidis (hereinafter, S. epidermidis, KCTC 1917) and they
were used from the Korean Collection for Type Cultures (KCTC) at Korea
Research Institute of Bioscience & Biotechnology (KRIBB). The culture was
performed in nutrient broth (DB, USA) at 37 °C, each strain was inoculated in 7
ml of liquid medium and activated while subcultured at regular time intervals.
In order to measure the antibacterial activity of the extract prepared
according to the Example 1, a paper disc method that can clearly identify the
antibacterial activity was performed. A platinum (wire) loop of each strain
cultured in a plate medium was incubated for 18 to 24 hours in 7 ml of liquid
medium, and then 0.1 ml of the bacterial solution was inoculated in 10 ml of the
liquid medium and incubated for 3 to 6 hours, and inoculated to be about 107
cells of 1 bacteria of plate medium and evenly spread. The sterilized filter paper
disc (Tokyo, 8 mm, Japan) was placed on a solid plate medium, and then the
extract sample was absorbed so as to a concentration of 0.05 ml/disc by
concentrations and incubated at 37 °C for 18 to 24 hours, then the diameter of
the clear zone (mm) around the disc was measured.
FIG. 2 is an image showing the antibacterial activity of the Mangifera
indica irwin seed extract according to an experimental example of the present invention, and Table 2 is a results measuring clear zone diameter of each strain according to the Mangifera indica irwin seed extract treatment concentration and showing as mean±SD of three measured values.
[Table 2]
Concentration Diameter of clear zone (mm) strain (mg/ml) Hot water extract (WE) Ethanol extract (EE) 1 - (no clear zone) 9.0±0.0 5 - 10.0±0.0 E.coli 10 - 11.0±0.9 20 - 13.0±1.1 1 - 8.0±0.0 5 - 9.0±0.7 S.epidermidis 10 - 9.5±11 20 - 11.5±0.8 1 - 8.0±0.0 5 - 10.0±0.3 S.aureus 10 - 11.0±0.2 20 - 13.0±1.0
Referring to FIG. 2 and Table 2, the activity was not shown in the hot
water extract (WE), but 70% ethanol extract (EE) showed a tendency to
increase the antibacterial activity as the treatment concentration of the extract
increases. Most of all strains were found to have a relatively high antibacterial
activity, especially in E. Coli was found to be a big clear zone of 9.0 - 13.0 mm.
Therefore, it was confirmed that the Mangifera indica irwin seed extract
according to an embodiment of the present invention has a high possibility as
an antibacterial material against skin flora that causes skin inflammatory
diseases.
<Experimental Example 4> Confirmation of cell viability
RAW264.7 cells, which are mouse macrophage lines obtained from
Korea Cell Line Bank were cultured in Dulbecco's Modified Eagle Medium
(hereinafter, DMEM, HyClone, USA) containing 10% fetal bovine serum (FBS,
HyClone, USA), 100 U/ml penicillin/streptomycin (HyClone, USA) in an
incubator at 37 °C, 5% C02. The amount of cells was cultured to grow about 80
90%.
MTT assay was performed to confirm the survival rate on the cells when
the extract prepared according to the Example 1 was exposed to the cells. First,
a 96 well plate was dispensed to 1x10 5 cells/well, and after 24 hours, an extract
sample dissolved in media was added to adjust each concentration. 40 pl of 3
(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, 2.5 mg/ml)
was added to each wells, and then reacted in an incubator for 4 hours to
remove supernatant. Dimethyl sulfoxide (DMSO) was mixed and completely
dissolved in 100 pl aliquots and then measured at 540 nm using a
spectrophotometer (Benchmark PLUS Bio-Rad).
FIG. 3 is a graph measuring cell viability according to an experimental
example of the present invention. TMD means the Mangifera indica irwin seed
hot water extract, TME means the Mangifera indica irwin seed ethanol extract,
and the graph shows the average of three measured values (*** p <0.001).
Referring to FIG. 3, the Mangifera indica irwin seed extract showed a
viability of 95% or more at 100 pg/ml and a cell viability of 50% or less from the concentration of 500 pg/ml. Accordingly, the next experiment was carried out at a concentration of 5, 10, 50, 100pg/ml.
<Experimental Example 5> Confirmation of nitric oxide (NO)
inhibition activity
Nitric oxide (hereafter, NO) was determined by measuring the amount of
NO in the supernatant of the cells as nitrite and nitrate. A griess reagent which
is safe form after reduction to nitrite for nitrate was used, and when the
confluence of RAW264.7 cells in a 96 well plate (5x104 cells/well) was 80%, it
was washed twice with phosphate buffer saline (PBS) and the cells were
incubated for 18 hours using a serum-free medium. Thereafter, the extract
samples were treated by concentration and incubated for 1 hour, followed by
incubation for 24 hours by treating with lipopolysaccharide (LPS) 1pg/ml. After
taking the supernatant of the culture and reacted with the griess reagent, the
absorbance was measured at 540 nm with an ELISA reader to express the NO
production rate as a percentage.
FIG. 4 is a graph measuring the nitric oxide (NO) production rate
according to an experimental example of the present invention. TMD means the
Mangifera indica irwin seed hot water extract, TME means the Mangifera indica
irwin seed ethanol extract, and the graph shows the average of three measured
values (*** p <0.001).
Referring to FIG. 4, the NO production rate of the cells rapidly increased
to 6.758±0.18 mM/ml when LPS was treated, but when the Mangifera indica irwin seed extract sample was treated, the NO production rate was found to decrease in concentration-dependent manner. Especially, at 100 pg/ml, hot water extract decreased to 4.65±0.29 mM/ml and ethanol extract decreased to
4.32 ±1.03 mM/mi, which found to decrease the most.
<Experimental Example 6> Confirmation of expression of
proinflammatoryfactors
After suspending RAW264.7 cells in DMEM containing 10% FBS, 3 ml
were dispensed in 6-well plates so as to be 2x10 6 cells/well and incubated in an
incubator at 37 °C, 5% C02 for 24 hours. After exchanging with fresh DMEM
medium, the cells were incubated with a stimulant LPS for 2 hours, and then the
extracts were cultured by treating the cells by concentration. After 24 hours, the
medium was removed to measure protein expression, washed with cold PBS,
and protein was extracted from the cell lysates by adding Protease
& Phosphatase Single-Use inhibitor Cocktail 10OX to RIPA buffer. Proteins were
quantified (BCA protein kit), electrophoresed on 10% SDS-PAGE, transferred to
membranes and then blocked with 5% skimmed milk for 1 hour.
Protein expression of COX-2 was confirmed using enhanced
chemiluminescence (ECL) solution (Thermo, USA). p-actin (Santa Cruz Biotechnology, Santa Cruz, California, USA) diluted in 1: 1000 was used as an
internal control.
FIG. 5 is a result confirming the expression of proinflammatory factors
according to an experimental example of the present invention.
Referring to FIG. 5, as a result of treating the Mangifera indica irwin
seed extract at a concentration of 5 to 100 pg/ml in RAW264.7 cells and
measuring the expression level of COX-2 protein, the expression level of the
protein in the LPS alone-treated group and treated group at each concentration
were increased, compared with the LPS-untreated group and when the extract
was treated, it was confirmed that the amount of COX-2 expression was
decreased in the concentration-dependent manner.
The Mangifera indica irwin seed extract according to the present
invention has antimicrobial activity, anti-inflammatory activity and antioxidant
activity against skin flora that causes skin inflammatory diseases and thus is
can be used as a cosmetic composition for anti-aging or skin inflammation
improvement or a composition of skin external preparation, and by using it, it
can effectively improve skin inflammation and prevent aging.
In addition, it is the extract using the natural raw material and thus has
few side effects and can be used safely.
While the present invention has been particularly described with
reference to specific embodiments thereof, it is apparent that this specific
description is only a preferred embodiment and that the scope of the present
invention is not limited thereby to those skilled in the art. That is, the practical
scope of the present invention is defined by the appended claims and their
equivalents.
Claims (10)
1. A cosmetic composition comprising Mangifera indica irwin seed
extract as an active ingredient.
2. The cosmetic composition of claim 1, wherein the extract is extracted
with water, C1 to C4 alcohol or a mixed solvent thereof.
3. The cosmetic composition of claim 1, wherein the extract has an
antimicrobial activity against at least one skin flora selected from the group
consisting of Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and
Staphylococcus epidermidis (S. epidermidis).
4. The cosmetic composition of claim 1, wherein the extract has anti
inflammatory activity by reducing production of nitric oxide (NO) or inhibiting
expression of COX-2 (cyclooxygenase-2).
5. The cosmetic composition of claim 1, wherein the extract has an
antioxidant activity.
6. The cosmetic composition of claim 1, wherein the composition is for
anti-aging or skin inflammation improvement.
7. The cosmetic composition of claim 1, wherein the extract comprises
0.001 to 50 parts by weight based on 100 parts by weight of total cosmetic
composition.
8. The cosmetic composition of claim 1, the composition has a
formulation of softening lotion, nourishing lotion, nourishing cream, essence or
pack.
9. A composition of skin external preparation for anti-aging or improving
skin inflammation comprising Mangifera indica irwin seed extract as an active ingredient.
10. A composition for inhibiting growth of skin flora comprising Mangifera
indica irwin seed extract as an active ingredient.
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| KR1020190105979A KR102359223B1 (en) | 2019-08-28 | 2019-08-28 | Cosmetic composition comprising mangifera indica irwin seed extract |
| KR10-2019-0105979 | 2019-08-28 |
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| AU2019236633A1 true AU2019236633A1 (en) | 2021-03-18 |
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| KR101872458B1 (en) | 2016-09-02 | 2018-06-28 | 주식회사 프롬바이오 | Food composition with the extract of Dichrostachys glomerata fruit and the extract of Irvingia gabonensis seed for improvement of obesity and hyperlipidemia |
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| KR20210025930A (en) | 2021-03-10 |
| AU2019236633B2 (en) | 2021-05-27 |
| KR102359223B1 (en) | 2022-02-07 |
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