COMPOSITION CONTAINING FUCOSTEROL FOR SKIN WHITENING OR MOISTURIZING
20170000714 ยท 2017-01-05
Assignee
Inventors
Cpc classification
A23V2200/318
HUMAN NECESSITIES
A23V2002/00
HUMAN NECESSITIES
A23L33/105
HUMAN NECESSITIES
A23V2002/00
HUMAN NECESSITIES
A61K31/575
HUMAN NECESSITIES
A23L33/11
HUMAN NECESSITIES
A61K9/0019
HUMAN NECESSITIES
A61K9/0014
HUMAN NECESSITIES
A23V2200/318
HUMAN NECESSITIES
A61K9/2054
HUMAN NECESSITIES
International classification
A23L33/11
HUMAN NECESSITIES
A61K31/575
HUMAN NECESSITIES
Abstract
The present invention relates to a novel use of fucosterol and, more specifically, to a composition for skin whitening or composition for skin moisturizing, which is characterized by containing fucosterol. Fucosterol of chemical formula 1 below has an excellent whitening effect of inhibiting melanin generation and tyrosinase activity, and thus can be used as an active ingredient of a cosmetic composition for whitening, a food composition, or a pharmaceutical composition. Furthermore, the fucosterol of chemical formula 1 below has an excellent moisturizing function by forming dead skin cell membranes, promoting the differentiation of dead skin cells, and generating natural moisturization factors, and thus can be used as an active ingredient of a cosmetic composition for moisturizing, a food composition, or a pharmaceutical composition
##STR00001##
Claims
1. A cosmetic composition for skin whitening, the cosmetic composition comprising fucosterol represented by the following Chemical Formula 1 as an active ingredient: ##STR00005##
2. A food composition for skin whitening, the food composition comprising fucosterol represented by the following Chemical Formula 1 as an active ingredient: ##STR00006##
3. A pharmaceutical composition for skin whitening, the pharmaceutical composition comprising fucosterol represented by the following Chemical Formula 1 as an active ingredient: ##STR00007##
4. A cosmetic composition for skin moisturizing the cosmetic composition comprising fucosterol represented by the following Chemical Formula 1 as an active ingredient: ##STR00008##
5. A food composition for skin moisturizing, the food composition comprising fucosterol represented by the following Chemical Formula 1 as an active ingredient: ##STR00009##
6. A pharmaceutical composition for skin moisturizing, the pharmaceutical composition comprising fucosterol represented by the following Chemical Formula 1 as an active ingredient: ##STR00010##
7. A method of improving a skin condition, the method comprising administrating a composition including fucosterol represented by the following Chemical Formula 1 as an active ingredient to a subject, wherein the skin condition improvement is skin whitening or skin moisturizing: ##STR00011##
Description
DESCRIPTION OF DRAWINGS
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BEST MODE
[0068] Hereinafter, the present invention will be described in detail by the following Examples. However, the following Examples are just provided for illustrative purposes in order to help in the understanding for the present invention and the scope of the present invention is not limited thereto.
[0069] In all test results below, the activity analysis was performed repetitively three times or more, and the results were represented by meanstandard deviation. Statistical analysis used an ANOVA analysis method (Scheff test) and when a *P value was equal to or less than 0.05 or a .sup.##P value and **P value was equal to or less than 0.01, the values were determined to be statistically significant.
Example 1
Extraction, Isolation, and Purification of Fucosterol
[0070] 500 g of dried Silvetia siliquosa was grinded by a mixer, and a sample of the grinded Silvetia siliquosa was put in n-hexane in four times volume and macerated for 48 hrs at room temperature to be extracted. The extracted sample was filtered with a Whatman No. 2 filter, the filtered extract was concentrated by a vacuum rotary concentrator, and then a solvent component was removed to obtain about 15.0 g of a Silvetia siliquosa n-hexane extract. 15 g of n-hexane soluble extract was loaded on a silicagel open column (70-230 mesh, Merck&Co., Whitehouse Station, N.J., USA) and split by using a solvent system of mixing hexane and ethyl acetate. According to the split order, 40 lower fractions were divided with a concentration gradient and then the fucosterol which was a compound represented by the following Chemical Formula 1 was isolated (210 mg) from 10-th to 30-th fractions among the fractions.
##STR00004##
Example 2
Safety Effect According to Fucosterol Treatment in Melanoma Cells
[0071] B16F10 melanoma cells were incubated a dulbecco's modified eagle's media (DMEM) medium including 10% fetal bovine serum and then added in a 24-well plate with 2.510.sup.4 cell/mL (the final volume of 1 mL). Fucosterol of 0.1, 1, 5, 10, and 20 M was treated in the B16F10 melanoma cells, respectively. After treatment of 48 hrs, the medium was removed and a 0.5 mg/mL MTT solution was put in each well by 0.3 mL and the B16F10 melanoma cells were incubated in an incubator for 4 hrs. After 4 hrs, the MTT solution was removed and generated formazan was dissolved by adding dimetyl sulfoxide (DMSO) and then absorbance was measured at 570 nm and the result was illustrated in
[0072] As illustrated in
Example 3
Inhibitory Effect on Melanin Generation According to Fucosterol Treatment in Melanoma Cells
[0073] B16F10 melanoma cells were incubated in a DMEM medium including 10% fetal bovine serum and then added in a 6-well plate with 2.510.sup.5 cell/mL (the final volume of 3 mL). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium including 200 nM -MSH was treated at 5, 10, and 20 M concentrations, respectively. After 72 hrs, the incubated medium was removed from the 6-well plate and 0.25% trypsin-ethylenediaminetetraacetate (trypsin-EDTA) solution was treated to collect a cell pellet, and the cell pellet was transferred to a 1.5 mL tube and centrifuged for 10 min at 10,000 rpm to remove a supernatant. The obtained pellet was dried at 60 C. and added with 1N NaOH 100 L to lyse melanin in the cells. This solution was diluted with phosphate buffered saline (PBS) and then the melanin content in a sample treatment group was calculated by measuring absorbance at 405 nm by an ELISA reader (Versa max, Sunnyvale, Calif., USA). In this case, cells without the fucosterol were set to a control group and the melanin generation degree was measured in the control group and cells treated with the fucosterol. According to Equation 1 below, the melanin content compared with the control group was calculated and the result was illustrated in
Melanin content (%) compared with control group=absorbance of each test material/absorbance of control group100[Equation 1]
[0074] As illustrated in
Example 4
Microscopic Observation of Inhibitory Effect on Melanin Generation According to Fucosterol Treatment in Melanoma Cells
[0075] B16F10 melanoma cells were incubated in a DMEM medium including 10% fetal bovine serum and then added in a 6-well plate with 2.510.sup.5 cell/mL (the final volume of 3 mL). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium including 200 nM -MSH was treated at 5 and 20 M of concentrations, respectively. After 72 hrs, the incubated medium was removed from the 6-well plate and the B16F10 melanoma cells were photographed by using a microscope (Olympus IX71 Model), and the result was illustrated in
[0076] As illustrated in
Example 5
Inhibitory Effect on Tyrosinase Activity According to Fucosterol Treatment in Melanoma Cells
[0077] B16F10 melanoma cells were incubated in a DMEM medium including 10% fetal bovine serum and then added in a 6-well plate with 2.510.sup.5 cell/mL (the final volume of 3 mL). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium including 200 nM -MSH was treated at 5, 10, and 20 M concentrations, respectively. After 48 hrs, the incubated medium was removed from the 6-well plate and PBS including 1% Triton X-100 was added to collect the cells. The collected cells were centrifuged for 10 min and 150 L of a supernatant was put in a 96-well plate and 50 L of L-3,4-dihydroxyphenylalanine (L-DOPA) was added. With respect to an amount of dopachrome generated after incubated for 30 min at 37 C., absorbance was measured at 475 nm by using a microplate reader (Versa max, Sunnyvale, Calif., USA). In this case, cells without the fucosterol were set to a control group and the tyrosinase activity degree was measured in the control group and cells treated with the fucosterol. According to Equation 2 below, the tyrosinase activity compared with the control group was calculated and the result was illustrated in
Tyrosinase activity (%) compared with control group=Reaction absorbance of each test material/Reaction absorbance of control group100[Equation 2]
[0078] As illustrated in
Example 6
Reduction Effect on Tyrosinase Protein Expression According to Fucosterol Treatment in Melanoma Cells
[0079] B16F10 melanoma cells were incubated in a DMEM medium including 10% fetal bovine serum and then added in a 6-well plate with 2.510.sup.5 cell/mL (the final volume of 3 mL). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium including 200 nM -MSH was treated at 5 and 20 M of concentrations, respectively. The cells after 24 hrs were lysed with a NP40 buffer solution including proteinase inhibitor cocktail and the protein amount extracted from the cells was quantified by using a Bio-Rad protein assay dye (Bradford) reagent. The quantified protein was boiled for 5 min and isolated by electrophoresis with 10% SDS-PAGE to transfer the isolated proteins to a nitrocellular membrane. A primary antibody of tyrosinase was diluted in 2.5% bovine serum albumin at a ratio of 1:1000 and reacted with the protein transferred to the nitrocellular membrane for 20 hrs at room temperature. The primary antibody was reacted and the nitrocellular membrane was washed by using tris-buffer saline tween 20 (TBST) three times for 10 min. After washing, a secondary antibody (anti-goat horseradish) recognizing the primary antibody was diluted in 2.5% bovine serum albumin to be 1:5000 and reacted with the nitrocellular membrane for 2 hrs at room temperature and then washed three times by 10 min by using the TBST. A protein band was colored by using ECL western blotting detection reagents (Amersham, Tokyo, Japan) and the colored protein band was verified by using a G:BOX EF imaging system (Syngene, Cambridge, UK). As the measured result, the protein expression of tyrosinase was verified and the loading amount of the protein was constant by -tubulin.
[0080] As illustrated in
Example 7
Reduction Effect on Protein Expression of Tyrosinase-Related TRP-1 and TRP-2 According to Fucosterol Treatment in Melanoma Cells
[0081] B16F10 melanoma cells were incubated in a DMEM medium including 10% fetal bovine serum and then added in a 6-well plate with 2.510.sup.5 cell/mL (the final volume of 3 mL). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium including 200 nM -MSH was treated at 5 and 20 M of concentrations, respectively. The cells after 24 hrs were lysed with a NP40 buffer solution including proteinase inhibitor cocktail and the protein amount extracted from the cells was quantified by using a Bradford reagent. The quantified protein was boiled for 5 min and isolated by electrophoresis with 10% SDS-PAGE to transfer the isolated proteins to a nitrocellular membrane. A primary antibody of TRP-1 and TRP-2 was diluted in 2.5% bovine serum albumin at a ratio of 1:1000 and reacted with the protein transferred to the nitrocellular membrane for 20 hrs at room temperature. The primary antibody was reacted and then the nitrocellular membrane was washed three times by using the TBST for 10 min. After washing, a secondary antibody (anti-goat horseradish) recognizing the primary antibody was diluted in 2.5% bovine serum albumin to be 1:5000 and reacted with the nitrocellular membrane for 2 hrs at room temperature and then washed three times by 10 min by using the TBST. A protein band was colored by using ECL western blotting detection reagents (Amersham, Tokyo, Japan) and the colored protein band was verified by using a G:BOX EF imaging system (Syngene, Cambridge, UK). As the measured result, the protein expression of TRP-1 and TRP-2 was verified and it was shown that the loading amount of the protein was constant by -tubulin.
[0082] As illustrated in
Example 8
Reduction Effect on mRNA Expression of MITF According to Fucosterol Treatment in Melanoma Cells
[0083] B16F10 melanoma cells were incubated in a DMEM medium including 10% fetal bovine serum and then added in a 6-well plate with 2.510.sup.5 cell/mL (the final volume of 3 mL). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium including 200 nM -MSH was treated at 5 and 20 M of concentrations, respectively. After 24 hrs, the total RNA was isolated by using a TRIzol reagent (Invitrogen, Carlsbad, Calif., USA). The isolated total RNAs were quantified by using a nano-drop (ND-1000). The quantified RNA was synthesized to cDNA by using reverse transcriptase and a PCR machine (Applied biosystems, Foster city, CA, USA) and then the PCR was performed with specific primers below.
TABLE-US-00001 GAPDH (SEQIDNO:1) Forwardprimer; 5-ACCACAGTCCATGCCATCAC-3 (SEQIDNO:2) Reverseprimer; 5-CCACCCGAGCCACATCGCTC-3 MITF (SEQIDNO:3) Forwardprimer; 5-AGTCAACCTCTGAAGAGCA-3 (SEQIDNO:4) Reverseprimer; 5-CGTGTTCATACCTGGGCACT-3
[0084] As the PCR result, the amplified cDNA was isolated with a 1.5% agarose gel by electrophoresis and cDNA bands were verified by using a G:BOX EF imaging system (Syngene, Cambridge, UK), and the result was illustrated in
[0085] As illustrated in
Example 9
Effect on Cornified Envelope Formation According to Fucosterol Treatment in HaCaT Human Keratinocytes
[0086] HaCaT human keratinocytes were incubated in a Dulbecco's Modified Eagle's Media (DMEM) medium including 10% fetal bovine serum and then put in a 6-well plate with 210.sup.5 cell/ml (the final volume of 3 ml). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium was treated at 5, 10, and 20 M concentrations, respectively. After incubation for 96 hrs, the incubated medium was removed from the 6-well plate to collect a cell pellet, and the cell pellet was transferred to a 1.5 mL tube and centrifuged for 10 min at 10,000 rpm to remove a supernatant. The obtained pellet was added with a Tris buffer solution including 2% sodium dodecyl sulfate (SDS) and 20 mM dithiothreitol (DTT) and boiled, and the absorbance was measured at 340 nm to evaluate the effect on cornified envelope formation. According to Equation 1 below, the formation degree of the cornified envelope compared with the control group was calculated and the result was illustrated in
Cornified envelope formation amount (%) compared with control group=absorbance of each test material/absorbance of control group100[Equation 3]
[0087] As illustrated in
Example 10
Effect on Increase in mRNA Expression Levels of Loricrin, Involucrin, and Transglutaminase According to Fucosterol Treatment in HaCaT Human Keratinocytes
[0088] HaCaT human keratinocytes were incubated in a Dulbecco's Modified Eagle's Media (DMEM) medium including 10% fetal bovine serum and then put in a 6-well plate with 210.sup.5 cell/ml (the final volume of 3 ml). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium was treated at 5, 10, and 20 M concentrations, respectively. After 24 hrs, the total RNA was collected by using a TRIzol reagent (Invitrogen, Carlsbad, Calif., USA) and reverse-transcribed and then a reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed below. First, for cDNA synthesis, the RNA was reverse-transcribed by reverse transcriptase. The RT-PCR was performed by specific primers below and the result was illustrated in
TABLE-US-00002 GAPDH (SEQIDNO:5) Forwardprimer; 5-TGACCTTGGCCAGGGGTGCT-3 (SEQIDNO:6) Reverseprimer; 5-CCACCCGAGCCACATCGCTC-3 Loricrin (SEQIDNO:7) Forwardprimer; 5-GGGTACCACGGAGGCGAAGGA-3 (SEQIDNO:8) Reverseprimer; 5-ACTGAGGCACTGGGGTTGGGA-3 Involucrin (SEQIDNO:9) Forwardprimer; 5-GGGGCAGCTGAAGCACCTGG-3 (SEQIDNO:10) Reverseprimer; 5-GAGACGGGCCACCTAGCGGA-3 Transglutaminase (SEQIDNO:11) Forwardprimer; 5CTTCCGTCTGCGCACCCCAG-3 (SEQIDNO:12) Reverseprimer; 5-AGGCACAAACGACTGGCGCA-3
[0089] As illustrated in
Example 11
Effect on Increase in Protein Expression Levels of Loricrin, Involucrin, and Transglutaminase According to Fucosterol Treatment in HaCaT Human Keratinocytes
[0090] HaCaT human keratinocytes were incubated in a Dulbecco's Modified Eagle's Media (DMEM) medium including 10% fetal bovine serum and then put in a 6-well plate with 210.sup.5 cell/ml (the final volume of 3 ml). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium was treated at 5, 10, and 20 M concentrations, respectively. After 24 hrs, the HaCaT human keratinocytes were lysed with a NP40 buffer solution including proteinase inhibitor cocktail. The protein amount extracted from the HaCaT human keratinocytes was quantified by using a Bradford method. The sample was boiled for 5 min and then the same amount of protein (20 g) was isolated by electrophoresis with 10% SDS-PAGE. The isolated proteins after electrophoresis were transferred to a nitrocellular membrane and western blot was performed. A primary antibody was reacted and the nitrocellular membrane was washed three times by using Tris-buffer saline tween20 (TBST) for 10 min. In this case, the kind and the dilution of the primary antibody used in the present invention is 1:1000. In a secondary antibody reaction, a secondary antibody was put in the membrane performing the primary antibody reaction and reacted for 2 hrs at room temperature. In this case, a dilution of the secondary antibody was 1:5000. Protein bands were colored by using ECL western blotting detection reagents (Amersham, Tokyo, Japan). The protein expression of loricrin, involucrin, and transglutaminase was verified and the loading amount of the protein was constant by -tubulin.
[0091] As illustrated in
Example 12
Effect on Increase in mRNA Expression Levels of Filaggrin and Caspase 14 According to Fucosterol Treatment in HaCaT Human Keratinocytes
[0092] HaCaT human keratinocytes were incubated in a Dulbecco's Modified Eagle's Media (DMEM) medium including 10% fetal bovine serum and then put in a 6-well plate with 210.sup.5 cell/ml (the final volume of 3 ml). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium was treated at 5, 10, and 20 M concentrations, respectively. After 24 hrs, the total RNA was collected by using a TRIzol reagent (Invitrogen, Carlsbad, Calif., USA) and reverse-transcribed and then a reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed below. First, for cDNA synthesis, the RNA was reverse-transcribed by reverse transcriptase. The RT-PCR was performed by specific primers below and the result was illustrated in
TABLE-US-00003 GAPDH (SEQIDNO:5) Forwardprimer; 5-TGACCTTGGCCAGGGGTGCT-3 (SEQIDNO:6) Reverseprimer; 5-CCACCCGAGCCACATCGCTC-3 Filaggrin (SEQIDNO:13) Forwardprimer; 5-AGTGCACTCAGGGGGCTCACA-3 (SEQIDNO:14) Reverseprimer; 5-CCGGCTTGGCCGTAATGTGT-3 Caspase14 (SEQIDNO:15) Forwardprimer; 5-CGGGACTCACAACCAAAGGA-3 (SEQIDNO:16) Reverseprimer; 5-GGGTCCCTTTGTTCTCCTCG-3
[0093] As illustrated in
Example 13
Effect on Increase in Protein Expression Levels of Filaggrin and Caspase 14 According to Fucosterol Treatment in HaCaT Human Keratinocytes
[0094] HaCaT human keratinocytes were incubated in a Dulbecco's Modified Eagle's Media (DMEM) medium including 10% fetal bovine serum and then put in a 6-well plate with 210.sup.5 cell/ml (the final volume of 3 ml). After being incubated for 24 hrs, the medium was removed and fucosterol which was dissolved in the DMEM medium was treated at 5, 10, and 20 M concentrations, respectively. After 24 hrs, the HaCaT human keratinocytes were lysed with a NP40 buffer solution including proteinase inhibitor cocktail. The protein amount extracted from the HaCaT human keratinocytes was quantified by using a Bradford method. The sample was boiled for 5 min and then the same amount of protein (20 g) was isolated by electrophoresis with 10% SDS-PAGE. The isolated proteins after electrophoresis were transferred to a nitrocellular membrane and western blot was performed. A primary antibody was reacted and the nitrocellular membrane was washed three times by using Tris-buffer saline tween20 (TBST) for 10 min. In this case, the kind and the dilution of the primary antibody used in the present invention is 1:1000. In a secondary antibody reaction, a secondary antibody was put in the membrane performing the primary antibody reaction and reacted for 2 hrs at room temperature. In this case, a dilution of the secondary antibody was 1:5000. Protein bands were colored by using ECL western blotting detection reagents (Amersham, Tokyo, Japan). The protein expression of filaggrin and caspase 14 was verified and it was shown that the loading amount of the protein was constant by -tubulin.
[0095] As illustrated in
[0096] Formulation Examples of the cosmetics for skin whitening including the fucosterol according to the present invention as an active ingredient are described, but the present invention is not limited thereto but will be described in detail. The cosmetic compositions for skin whitening of Formulation Examples 1 to 6 having the fucosterol having the excellent skin whitening method according to composition ingredients and composition ratios below were prepared by a general method.
Formulation Example 1
Cosmetics
[0097] <1-1> Nutrition Lotion (Milky Lotion)
[0098] The nutrition lotion was prepared by using the fucosterol of Example 1 according to a general method with a formulation ratio of the nutrition lotion in Table 1 below.
TABLE-US-00004 TABLE 1 Formulation Example 1-1 Combined ingredients (wt %) Fucosterol 2.0 Squalane 5.0 Wax 4.0 Polysorbate 60 1.5 Sorbitan Sesquioleate 1.5 Liquid paraffin 0.5 Caprylic/Capric Triglyceride 5.0 Glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Triethanolamine 0.2 Preservative, dye, flavoring Suitable amount Purified water to 100
[0099] <1-2> Emollient (Skin Lotion)
[0100] The emollient was prepared by using the fucosterol of Example 1 according to a general method with a formulation ratio of the emollient in Table 2 below.
TABLE-US-00005 TABLE 2 Formulation Example 1-2 Combined ingredients (wt %) Fucosterol 2.0 Glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 PEG 12 nonylphenylether 0.2 Polysorbate 80 0.4 Ethanol 10.0 Triethanolamine 0.1 Preservative, dye, flavoring Suitable amount Purified water to 100
[0101] <1-3> Nourishing Cream
[0102] The nourishing cream was prepared by using the fucosterol of Example 1 according to a general method with a formulation ratio of the nourishing cream in Table 3 below.
TABLE-US-00006 TABLE 3 Formulation Example 1-3 Combined ingredients (wt %) Fucosterol 2.0 Polysorbate 60 1.5 Sorbitan Sesquioleate 0.5 PEG60 hydrogenated castor oil 2.0 Liquid paraffin 10 Squalane 5.0 Caprylic/Capric Triglyceride 5.0 Glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservative Suitable amount Dye Suitable amount Flavoring Suitable amount Purified water to 100
[0103] <1-4> Massage Cream
[0104] The massage cream was prepared by using the fucosterol of Example 1 according to a general method with a formulation ratio of the massage cream in Table 4 below.
TABLE-US-00007 TABLE 4 Formulation Example 1-4 Combined ingredients (wt %) Fucosterol 1.0 Wax 10.0 Polysorbate 60 1.5 PEG 60 hydrogenated castor oil 2.0 Sorbitan Sesquioleate 0.8 Liquid paraffin 40.0 Squalane 5.0 Caprylic/Capric Triglyceride 4.0 Glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservative, dye, flavoring Suitable amount Purified water to 100
[0105] <1-5> Pack
[0106] The pack was prepared by using the fucosterol of Example 1 according to a general method with a formulation ratio of the pack in Table 5 below.
TABLE-US-00008 TABLE 5 Formulation Example 1-5 Combined ingredients (wt %) Fucosterol 1.0 Polyvinylalcohol 13.0 Sodium carboxymethyl 0.2 cellulose Glycerin 5.0 Allantoin 0.1 Ethanol 6.0 PEG 12 nonylphenylether 0.3 Polysorbate 60 0.3 Preservative, dye, flavoring Suitable amount Purified water to 100
[0107] <1-6> Gel
[0108] The gel was prepared by using the fucosterol of Example 1 according to a general method with a formulation ratio of the gel in Table 6 below.
TABLE-US-00009 TABLE 6 Formulation Example 1-6 Combined ingredients (wt %) Fucosterol 0.5 Sodium ethylene diamine 0.05 acetate Glycerin 5.0 Carboxyvinyl polymer 0.3 Ethanol 5.0 PEG 60 hydrogenated castor oil 0.5 Triethanolamine 0.3 Preservative, dye, flavoring Suitable amount Purified water to 100
Formulation Example 2
Food
[0109] <2-1> Preparation of Health Food
[0110] The health food may be prepared by mixing fucosterol 1000 mg of [Example 1], vitamin A acetate 70 g, vitamin E 1.0 mg, vitamin B1 0.13 mg, vitamin B2 0.15 mg, vitamin B6 0.5 mg, vitamin B12 0.2 g, vitamin C 10 mg, biotin 10 g, nicotinic acid amide 1.7 mg, folic acid 50 g, calcium pantothenate 0.5 mg, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, monopotassium phosphate 15 mg, dicalcium phosphate 55 mg, potassium citrate 90 mg, calcium carbonate 100 mg, and magnesium chloride 24.8 mg, and the combined ratio may be randomly modified. The ingredients are mixed according to a general method of preparing the health food to prepare granules and may be used in the preparation of a health food composition according to a general method.
[0111] <2-2> Preparation of Health Beverage
[0112] Fucosterol 1000 mg of [Example 1], citric acid 1000 mg, oligosaccharide 100 g, plum extract 2 g, and taurine 1 g are added with purified water, mixed according to a general method of preparing a health beverage of a total 900 mL, stirred and heated for about 1 hr at 85 C., and then the prepared solution is filtered, obtained in a sterilized container of 2 L, and refrigerated after sealing sterilization to be used for preparation of a health food composition.
[0113] <2-3> Chewing Gum
[0114] The chewing gum was prepared by mixing 0.1 wt % of the fucosterol of [Example 1] with gum base 20 wt %, sugar 76.9 wt %, flavoring 1 wt %, and water 2 wt % according to a general method.
[0115] <2-4> Candy
[0116] The candy was prepared by mixing 0.1 wt % of the fucosterol of [Example 1] with sugar 60 wt %, starch syrup 39.8 wt %, and flavoring 0.1 wt % according to a general method.
[0117] <2-5> Biscuit
[0118] The biscuit was prepared by mixing 0.1 wt % of the fucosterol of [Example 1] with first-class soft flour 25.59 wt %, first-class medium flour 22.22 wt %, refined sugar 4.80 wt %, salt 0.73 wt %, glucose 0.78 wt %, palm shortening 11.78 wt %, ammonium 1.54 wt %, sodium bicarbonate 0.17 wt %, sodium bisulfite 0.16 wt %, rice flour 1.45 wt %, vitamin B0.0001 wt %, milk flavor 0.04 wt %, water 20.6998 wt %, whole milk powder 1.16 wt %, substitute milk powder 0.29 wt %, moncalcium phosphate 0.03 wt %, spray salt 0.29 wt %, and spray oil 7.27 wt % according to a general method.
Formulation Example 3
Medicines
[0119] <3-1> Powder
[0120] 50 mg of the fucosterol of [Example 1] and 2 g of crystalline cellulose were mixed and then filled in an airtight bag to prepare the powder according to a general method of preparing the powder.
[0121] <3-2> Tablet
[0122] 50 mg of the fucosterol of [Example 1], 400 mg of crystalline cellulose, and 5 mg of magnesium stearate were mixed and then tableted to prepare the tablet according to a general method of preparing the tablet.
[0123] <3-3> Capsule
[0124] 30 mg of the fucosterol of [Example 1], 100 mg of whey protein, 400 mg of crystalline cellulose, and 6 mg of magnesium stearate were mixed and then filled in a gelatin capsule to prepare the capsule according to a general method of preparing the capsule.
[0125] <3-4> Injection
[0126] According to a general method of preparing injections, an active ingredient was dissolved in distilled water for injection, pH was adjusted to about 7.5, and then 100 mg of the fucosterol of [Example 1], distilled water for injection, and a pH adjusting agent were mixed and filled in an ample of 2 mL and sterilized to prepare the injection.
INDUSTRIAL APPLICABILITY
[0127] The fucosterol of chemical formula 1 has an excellent whitening effect of inhibiting melanin generation and tyrosinase activity to be prepared as a cosmetic composition for whitening, a food composition, or a pharmaceutical composition, and thus industrial applicability is high.
[0128] Further, the fucosterol of chemical formula 1 has an excellent moisturizing function by forming dead skin cell membranes, promoting the differentiation of dead skin cells, and generating natural moisturization factors to be prepared as a cosmetic composition for moisturizing, a food composition, or a pharmaceutical composition, and thus industrial applicability is high.