A composition containing ginger extract as an active ingredient that has skin whitening and skin elasticity improving effects.

Abstract

This invention provides a composition that inhibits the tyrosinase enzyme involved in melanin production and also inhibits the action of collagen-degrading enzymes, thereby promoting collagen synthesis and achieving sufficient effects of skin whitening and elasticity improvement without inducing skin irritation due to skin allergies. [Solution] A composition having skin whitening and skin elasticity improving effects, according to one embodiment of the present invention, contains ginger extract as an active ingredient, and the ginger extract can be extracted using methylene chloride.

Description

Technical Field

[0001] The present invention relates to a composition containing a ginger extract as an active ingredient and having effects of improving skin beauty and skin elasticity.

Background Art

[0002] The color of human skin is determined by the concentration and distribution of melanin inside the skin. Such concentration and distribution of melanin are also affected by environmental or physiological conditions such as ultraviolet rays of the sun, fatigue, stress, etc. in addition to genetic factors. [[ID=I4]]

[0003] The melanin pigment produced by melanocytes in human skin is a phenolic polymer substance having a complex form of black pigment and protein, and can play an important role in blocking skin damage caused by ultraviolet rays.

[0004] It has been reported that the action of tyrosinase present in melanocytes is most important for melanin biosynthesis. Tyrosinase can play an important role in the skin darkening process by converting tyrosine, which is a kind of amino acid, into DOPA and dopaquinone, which are intermediate products of melanin polymer production.

[0005] However, although the pathway by which melanin is produced is known, the mechanism that induces melanin synthesis, which is the stage before tyrosinase acts, has not yet been clarified in detail.

[0006] When such melanin synthesis occurs excessively in the skin, it can darken the skin tone and cause freckles, chloasma, etc.

[0007] Therefore, inhibiting the synthesis of melanin pigment in the skin can not only brighten skin tone and achieve skin whitening, but also improve hyperpigmentation such as age spots and freckles caused by ultraviolet rays, hormones, and genetic factors.

[0008] While substances with tyrosinase inhibitory activity, such as hydroquinone, ascorbic acid, kojic acid, and glutathione, can be used to whiten skin and improve hyperpigmentation, hydroquinone exhibits the desired whitening effect but has the problem of being highly irritating to the skin and requiring its use to be limited to extremely small amounts. Ascorbic acid is easily oxidized, causing problems such as discoloration and odor in cosmetics containing it, and kojic acid is unsafe in solution and complicates the cosmetic manufacturing process.

[0009] Furthermore, thiol compounds such as glutathione and cysteine ​​not only have a characteristic unpleasant odor, but also pose problems with transdermal absorption, and their glycosides and derivatives are highly polar, making them sometimes difficult to use as ingredients in cosmetics.

[0010] Furthermore, in the case of vitamin C, it is easily oxidized in aqueous solution, which is a problem because it does not provide sustained effects. For this reason, skin whitening compositions containing natural herbal extracts have been developed, but many of them are colored, posing problems in terms of formulation, and because the active ingredients have not been identified, there is a problem in that the same effect cannot be expected when manufacturing the product.

[0011] On the other hand, skin aging is known to be caused by a decrease in collagen content, which is a protein that makes up the majority of collagen in the dermal tissue of the skin. Since collagen provides tension and strength to the skin, a decrease in collagen is very closely related to skin aging and wrinkle formation.

[0012] Skin aging can be broadly classified into intrinsic aging due to physiological aging and photoaging due to prolonged exposure to ultraviolet radiation (UV). Repeated exposure to UV radiation induces denaturation and destruction of collagen fibers by increasing collagen-degrading enzymes, resulting in decreased skin elasticity and the formation of wrinkles.

[0013] Through this process, the inventors of the present invention have conducted extensive research and trial and error over many years to develop a composition for skin whitening and skin elasticity improvement that contains ginger extract as an active ingredient. This composition suppresses the tyrosinase enzyme involved in melanin production and inhibits the action of collagen-degrading enzymes, thereby promoting collagen synthesis and achieving sufficient effects of skin whitening and elasticity improvement without inducing skin irritation due to skin allergies. As a result, they have completed the present invention. [Overview of the project] [Problems that the invention aims to solve]

[0014] According to one embodiment of the present invention, a ginger extract extracted from ginger is used as an active ingredient in a cosmetic for skin whitening and elasticity improvement. By inhibiting the tyrosinase enzyme involved in the melanin production mechanism and inhibiting the action of collagen-degrading enzymes, collagen synthesis is promoted, thereby achieving sufficient whitening and elasticity improvement effects. Furthermore, a composition containing ginger extract as an active ingredient that does not induce skin irritation due to skin allergies can be provided.

[0015] On the other hand, other objectives not explicitly stated in the present invention may be further considered to the extent that they can be easily inferred from the detailed description and effects below. [Means for solving the problem]

[0016] According to one embodiment of the present invention, a composition having skin whitening and skin elasticity improving effects, which contains ginger extract as an active ingredient, contains ginger extract as an active ingredient, and the ginger extract can be extracted using methylene chloride.

[0017] The ginger extract can be prepared by extracting methylene chloride at 10°C for 50 hours, followed by 30 hours at 25°C.

[0018] The ginger extract can be included in an amount of 0.125 to 0.5 parts by weight per 100 parts by weight of the total. [Effects of the Invention]

[0019] According to one embodiment of the present invention, a composition containing ginger extract as an active ingredient, which has skin beautifying and skin elasticity improving effects, can be included as an active ingredient in cosmetics for skin whitening and skin elasticity improvement, and can provide sufficient effects for skin whitening and skin elasticity improvement, and can provide substantial and practical effects such as not inducing skin allergies.

[0020] On the other hand, even if an effect is not explicitly stated herein, the effects described in the following specification and any provisional effects expected by the technical features of the present invention shall be treated as described in the specification of the present invention.

[0021] The attached drawings are provided as illustrative examples for understanding the technical concept of the present invention, and this does not limit the scope of the rights of the present invention. [Brief explanation of the drawing]

[0022] [Figure 1] This graph shows the cell viability of the sample. [Figure 2] This graph shows the amount of extracellular melanin released from the sample. [Figure 3] This graph shows the inhibition rate of extracellular melanin production in the sample. [Figure 4]It is a graph showing the amount of intracellular melanin release in the sample. [Figure 5] It is a graph showing the inhibitory rate of intracellular melanin production in the sample. [Figure 6] It is a graph showing the total amount of melanin release in the sample. [Figure 7] It is a graph showing the inhibitory rate of total melanin production in the sample. [Figure 8] It is a graph showing the cell viability of the sample. [Figure 9] It is a graph showing the expression level of type I procollagen by the sample.

Mode for Carrying Out the Invention

[0023] When explaining the present invention, if it is determined that the details of related known functions would unnecessarily obscure the gist of the present invention as obvious matters to those skilled in the art, the detailed description thereof will be omitted.

[0024] The terms used in this application are merely used to explain a specific embodiment and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this application, terms such as "comprising" or "having" are intended to specify the presence of the features, numbers, steps, operations, components, parts, or combinations thereof described in this specification, and it should be understood that the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof is not precluded in advance.

[0025] Hereinafter, an embodiment of a skin whitening composition containing ginger extract according to the present invention as an active ingredient will be described in detail with reference to the accompanying drawings. When explaining with reference to the accompanying drawings, the same or corresponding components will be given the same drawing numbers, and duplicate explanations thereof will be omitted.

[0026] In this specification, "containing as an active ingredient" may mean that the ingredient is contained in an amount that is sufficient to significantly exhibit the effects described in this invention.

[0027] Ginger (Zingiber officinale) has the scientific name Zingiber officinale, and its root is scientifically known as Rhyzomus Zingiberus. The term "ginger" as commonly used can refer to the ginger root, which is the underground stem of the ginger plant.

[0028] Ayurvedic medicine and traditional Chinese herbal medicine (TCM) provide a good explanation of ginger's properties. The medicinal components of ginger are gingerol and shogaol, which are most abundant in raw ginger.

[0029] Ginger contains aromatic substances such as borneol, cineole, citral, phenol, and alkaloids as essential oil components. These remove unpleasant odors and tastes, such as fishy smells, and suppress central nausea. The pungent components gingerol and shogaol are known to have strong bactericidal effects against bacteria such as typhoid bacteria and cholera bacteria.

[0030] A ginger extract according to one embodiment of the present invention, which contains a ginger extract as an active ingredient in a composition having skin whitening and skin elasticity improving effects, can be produced by a ginger preparation step, a ginger steaming and drying step, and a ginger extract extraction step.

[0031] The ginger preparation step may be configured to remove impurities while washing the root portion of the ginger, and the ginger steaming and drying step may be a process of repeatedly steaming and drying.

[0032] In this step, the ginger steaming and drying process may be carried out at 70-100°C for 40-80 minutes, or more preferably, with steam at 90-95°C for 60-70 minutes.

[0033] Furthermore, in the ginger steaming and drying step, the drying process is carried out immediately after the steaming process is completed, and the drying process may be carried out at 65°C or below for 11 to 13 hours, or more preferably at 60°C for 60 minutes.

[0034] Thus, the ginger steaming and drying step may be configured to repeat the steaming and drying steps as a single continuous process four or more times, or more preferably, to repeat the steaming and drying steps as a single continuous process six times.

[0035] The ginger extract extraction step according to one embodiment of the present invention may be configured to extract ginger extract from ginger that has undergone the ginger steaming and drying step.

[0036] The method for extracting ginger extract in this invention is not particularly limited and can be used by methods commonly used in the art.

[0037] Methods for extracting ginger extract include hot water extraction, ultrasonic extraction, filtration, and reflux extraction. One of the above extraction methods may be selected and used, or two or more extraction methods may be selected and used in combination.

[0038] Furthermore, the ginger extract extraction step according to one embodiment of the present invention can be performed using a variety of extraction methods known in the art, and the extract can be obtained by extracting with water or an organic solvent, followed by concentration and drying.

[0039] As a solvent, water or anhydrous or aqueous lower alcohols having 1 to 4 carbon atoms (such as methanol, ethanol, propanol, isopropanol, butanol, etc.), a mixed solvent of the said lower alcohol and water, or dioxane and acetone, ethyl acetate, chloroform, methylene chloride, 1,3-butylene glycol, etc. can be used individually or in combination as extraction solvents.

[0040] Furthermore, in the ginger extract extraction step according to one embodiment of the present invention, extraction can preferably be performed using chloroform and methylene chloride.

[0041] In particular, in the ginger extract extraction step according to one embodiment of the present invention, extraction can most preferably be performed using methylene chloride.

[0042] Furthermore, in the ginger extract extraction step according to one embodiment of the present invention, a conventional method can be used with water or an organic solvent, for example, hot water extraction, high-pressure hot water extraction, ethanol extraction, methanol extraction, or organic solvent extraction can be performed.

[0043] In the case of hot water extraction, it may be carried out at 90-110°C, preferably at 100°C for 1-24 hours, and in the case of high-pressure hot water extraction, it may be carried out at 110-130°C, preferably at 120°C for 1-24 hours.

[0044] Extraction with ethanol, methanol, or propanol may be carried out at 60-90°C, preferably at 80°C for 1-24 hours, while extraction with organic solvents may be carried out at 18-40°C, preferably at 22-33°C for 50 hours.

[0045] In each of the above extraction methods, if extraction is performed outside the specified range of extraction temperature and time, the extraction efficiency may decrease or the extracted components may change. Therefore, it is preferable to perform the extraction within the specified range of extraction temperature and time.

[0046] In the ginger extract extraction step according to one embodiment of the present invention, the ginger extract obtained by the extraction method described above is a ginger concentrate and may be in the form of an emulsion, which is a highly viscous solution.

[0047] Alternatively, the obtained ginger extract can be macerated at room temperature, heated, and filtered to obtain a liquid, and the solvent can be further evaporated, spray-dried, or freeze-dried.

[0048] A ginger extract according to one embodiment of the present invention has tyrosinase inhibitory activity and can act to provide excellent skin whitening effects.

[0049] A composition having skin whitening and skin elasticity improving effects, containing ginger extract as an active ingredient according to one embodiment of the present invention, may contain ginger extract, which is the active ingredient of the present invention, in an amount of 0.05 to 20 parts by weight per 100 parts by weight of the composition.

[0050] In this case, if the amount of active ingredient is less than 0.05 parts by weight, the skin whitening effect may be significantly reduced, and if it exceeds 20 parts by weight, it may cause skin irritation or lead to formulation problems.

[0051] A composition having skin whitening and skin elasticity improving effects, containing ginger extract as an active ingredient according to one embodiment of the present invention, may preferably contain 0.125 to 4 parts by weight of ginger extract, which is the active ingredient of the present invention, per 100 parts by weight of the composition.

[0052] A cosmetic composition for skin whitening and skin elasticity improvement, based on a composition having skin whitening and skin elasticity improving effects that contains ginger extract according to one embodiment of the present invention as an active ingredient, may have dosage forms such as solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations, and sprays.

[0053] This invention, through the following examples, analyzes the components and whitening activity of ginger extract produced using solvents and extraction methods, and verifies that a composition containing the extracted ginger extract as an active ingredient is effective in whitening and improving skin elasticity.

[0054] The following describes preferred embodiments for understanding the present invention.

[0055] However, the following embodiments are merely for the purpose of understanding the present invention, and the present invention is not limited to the following embodiments.

[0056] Example 1. Production of ginger extract

[0057] I purchased 4 kg of ginger root from Andong, Gyeongsangbuk-do, peeled it, thoroughly washed it to remove all foreign matter, and then air-dried it at room temperature for 2-4 days.

[0058] Furthermore, the naturally dried ginger roots were steamed at 95°C for 70 minutes, then dried at 60°C for 60 minutes, and this steaming and drying process was repeated six times.

[0059] At this point, the ginger roots, which had undergone steaming and drying processes, were finely chopped and then put into a blender and ground into particles smaller than 1 mm.

[0060] The ginger root powder, which had been ground to particles smaller than 1 mm, was precipitated in hot water, methanol, ethanol, propanol, chloroform, and methylene chloride, respectively, and an extraction process was carried out.

[0061] Hot water extraction was performed at 100°C for 15 hours, high-pressure hot water extraction at 120°C for 12 hours, and extraction with ethanol, methanol, and propanol was performed at 80°C for 7 hours each.

[0062] Extraction with chloroform and methylene chloride was carried out at 10°C for 50 hours, followed by extraction at 25°C for 30 hours.

[0063] Furthermore, after the ginger extract extraction process was completed, each solvent was concentrated by rotary vacuum, then further concentrated by centrifugal vacuum, and the concentrate was freeze-dried to obtain the solid component.

[0064] Example 2. Measurement of the whitening activity of ginger extract

[0065] In Example 2, the degree to which tyrosinase activity was inhibited was investigated, and the whitening activity (Tyrosinase Inhibition Activity) was confirmed.

[0066] First, tyrosinase is known as an enzyme that synthesizes melanin from L-tyrosine and L-DOPA (L-3,4-dihydroxyphenylalanine) in melanocytes within melanosomes.

[0067] In Example 2, the solids of each ginger extract obtained in Example 1 by hot water extraction, high-pressure hot water extraction, methanol extraction, ethanol extraction, propanol extraction, chloroform extraction, and methylene chloride extraction were dissolved in dimethyl sulfoxide (DMSO) at low and high concentrations of 0.5 mg / ml and 20 mg / ml, and the inhibitory activity of tyrosinase was measured. The results are shown in Table 1 below.

[0068] [Table 1]

[0069] As shown in Table 1 above, the hot water extract showed tyrosinase inhibitory activity (%) of 40.11±1.01 at a low concentration of 0.5 mg / ml and 51.63±1.39 at a high concentration of 20 mg / ml.

[0070] The high-pressure hot water extract showed tyrosinase inhibitory activity (%) of 47.82±1.44 at a low concentration of 0.5 mg / ml, and 66.23±1.04 at a high concentration of 20 mg / ml.

[0071] The methanol extract showed tyrosinase inhibitory activity (%) of 70.91±1.77 at a low concentration of 0.5 mg / ml and 77.06±2.02 at a high concentration of 20 mg / ml.

[0072] The ethanol extract showed tyrosinase inhibitory activity (%) of 63.42±1.60 at a low concentration of 0.5 mg / ml, and 78.02±1.43 at a high concentration of 20 mg / ml.

[0073] The propanol extract showed tyrosinase inhibitory activity (%) of 67.02±1.60 at a low concentration of 0.5 mg / ml, and 79.66±1.38 at a high concentration of 20 mg / ml.

[0074] Chloroform extract showed tyrosinase inhibitory activity (%) of 80.01±1.70 at a low concentration of 0.5 mg / ml, and 98.44±1.61 at a high concentration of 20 mg / ml.

[0075] Methylene chloride extract showed tyrosinase inhibitory activity (%) of 96.94±1.70 at a low concentration of 0.5 mg / ml, and 99.02±1.58 at a high concentration of 20 mg / ml.

[0076] Upon investigation, chloroform extract and methylene chloride extract showed superior tyrosinase inhibitory activity, regardless of concentration, both at low and high concentrations, compared to other extracts.

[0077] In particular, methylene chloride extract was shown to exhibit significantly superior tyrosinase inhibitory activity even at lower concentrations than chloroform extract.

[0078] Ultimately, ginger extract extracted using methylene chloride as a solvent was shown to be the most effective for skin whitening, even at low concentrations, due to its excellent tyrosinase inhibitory activity.

[0079] Example 3. Melanin production inhibitory activity test of a composition containing ginger extract as an active ingredient.

[0080] In Example 3, a test was conducted to confirm the melanin production inhibitory activity of a sample (hereinafter referred to as "the sample") containing 2 parts by weight or less of ginger extract extracted using methylene chloride as a solvent as an active ingredient.

[0081] When examining the melanin production process in human skin, melanin is biosynthesized in melanosomes of melanocytes. Stimulation activates enzymes involved in melanin formation (Tyrosinase, TRP1, TRP2), and once melanin is formed by these enzymes, the formed melanin can move to keratinocytes via resinous protrusions and undergo coloration. Alpha-Melanocyte-stimulating hormones (α-MSH) are among the stimulating factors for melanocytes.

[0082] There are five stages to inhibiting melanin production: firstly, blocking the stimulating signals to melanocytes, with vitamin C compounds and vitamin E being typical whitening agents; secondly, suppressing the genes involved in tyrosinase formation, with retinol being a typical whitening agent; thirdly, inhibiting glycosylation, the stage for tyrosinase activation, with glucosamine being a typical whitening agent; fourthly, inactivating tyrosinase, with kojic acid, arbutin, and vitamin C compounds being typical whitening agents; and fifthly, blocking the migration to keratinocytes, with niacinamide being a typical whitening agent.

[0083] In Example 3, the effect of a sample containing 2 parts by weight or less of ginger extract extracted using methylene chloride as a solvent on melanin production in melanocytes induced by α-MSH (alpha-melanocyte-stimulating hormones), a melanocyte-stimulating holomon, was investigated in vitro.

[0084] A. Cell culture

[0085] B16-F10 melanoma cells were cultured in 150 mm cell culture dishes at 37°C in a 5% CO2 incubator using DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (P / S). Cells that reached confluence were maintained through subculturing using trypsin-EDTA.

[0086] I. Cell viability experiment (WST-1 assay)

[0087] Dehydrogenases present in mitochondria of living cells produce a chromogenic substance called formazan when exposed to tetrazolium salt (WST). By measuring this formazan, the number of living cells was determined as described below.

[0088] 2 × 10⁶ cells in a 96-well cell culture plate 3 B16-F10 cells were cultured for 24 hours in DMEM culture medium supplemented with 10% FBS and 1% P / S at a cell / well concentration. Then, the medium was replaced with DMEM medium containing the sample at different concentrations and cultured for 72 hours. After the cultured cells were added to DMEM medium containing 10% EZ-CYTOX and allowed to react for 2 hours, the absorbance was measured at 450 nm using a plate multi-reader to determine cell viability.

[0089] Cell viability (%) = (Absorbance of sample-treated group / Absorbance of untreated group) × 100

[0090] C. Experiments on melanin production inhibitory activity inside / outside cells

[0091] (1) Place B16-F10 cells into a 6-well cell culture plate in a 1 × 10⁶ 5 Cells were cultured for 24 hours in DMEM culture medium supplemented with 10% FBS and 1% P / S at a concentration of cells / well.

[0092] (2) 100 μM α-melanocyte-stimulating hormone (α-MSH) was diluted in DMEM / Modified medium to prepare DMEM / Modified medium containing 100 nM α-MSH.

[0093] (3) The sample and the positive control group, β-arbutin, were prepared by diluting them to different concentrations in DMEM / Modified medium (100 nM α-MSH).

[0094] (4) After removing the culture medium of the cells that had been cultured for 24 hours, the cells were washed with 2 mL of DPBS.

[0095] (5) Each sample, diluted to different concentrations, was treated with 3 mL of each solution and incubated at 37°C in a 5% CO2 incubator for 48 to 72 hours.

[0096] (6) After the culture was complete, the cells and culture medium were collected and centrifuged.

[0097] (7) The supernatant was dispensed into 96-well plates in 100 μL portions, and the amount of extracellular melanin released was measured by measuring the absorbance at 490 nm.

[0098] (8) Remove the remaining supernatant, wash the cells with 1 mL of DPBS, and then dry them at 40°C.

[0099] (9) 120 μL of 1N NaOH (10% DMSO) was added to the dried cells, and the cells were lysed by heating in a 60°C water bath for 1 hour, followed by centrifugation at 15000 rpm for 1 hour.

[0100] (10) The supernatant was dispensed into 96-well plates in 100 μL increments, and the amount of intracellular melanin production was measured by measuring the absorbance at 490 nm.

[0101] (11) The measured melanin amounts were corrected using the total protein amount obtained by the BCA (Bicinchronic acid assay) and the synthetic melanin standard curve.

[0102] (12) Melanin production inhibition rate

[0103] Melanin production inhibition rate (%) = 100 - [(Melanin production amount in the sample-treated group / Melanin production amount in the α-MSH-treated control group) × 100]

[0104] E. Bicinchoninic acid (BCA) assay

[0105] (1) The amount of protein was confirmed using the Pierce BCA Protein Assay Kit (23227, Thermo Scientific).

[0106] (2) The amount of melanin produced in the cells was measured, and the remaining supernatant was diluted by 1 / 10 with distilled water for each concentration to prepare the sample.

[0107] (3) Diluted samples were dispensed into 96-well plates in 20 μL portions according to concentration, 200 μL of Pierce BCA Protein Assay Reagent mixture was added and mixed, and after standing at 37°C for approximately 30 minutes, the absorbance was measured at 570 nm.

[0108] (4) The protein content was confirmed using BSA (bovine serum albumin) as a standard substance.

[0109] O. Data analysis and statistical processing

[0110] The statistical significance of all experimental results was confirmed using the t-test (independent t-test, two-tailed test, unequal variance assumption) function in Microsoft Office Excel 2007, and a p-value of less than 0.05 was considered statistically significant.

[0111] C. Experimental Results

[0112] (1) Effect on the survival rate of B16-F10 melanoma cells

[0113] Table 2 below shows the results of the cytotoxicity test on the samples, and Figure 1 attached is a graph showing the cell viability of the samples.

[0114] The effect of samples containing 2 parts by weight or less of ginger extract as an active ingredient on B16-F10 cell viability was investigated and is shown in Table 2 and Figure 1.

[0115] As a result, samples containing 2 parts by weight or less of ginger extract as an active ingredient showed a cell viability rate of 90% or more.

[0116] [Table 2]

[0117] (2) Effect of samples containing 2 parts by weight or less of ginger extract as an active ingredient on extracellular melanin release

[0118] The effects of samples containing β-arbutin and ginger extract as active ingredients (2 parts by weight or less), which served as a positive control group, on extracellular melanin production in B16-F10 melanoma cells were investigated.

[0119] Table 3 below shows the experimental results for the amount of extracellular melanin released from the sample, and Table 4 shows the experimental results for the inhibition rate of extracellular melanin production from the sample.

[0120] Furthermore, Figure 2 is a graph showing the amount of extracellular melanin released from the sample, and Figure 3 is a graph showing the inhibition rate of extracellular melanin production from the sample.

[0121] As shown in Tables 3 and 4 below, β-arbutin, the positive control group, was confirmed to inhibit extracellular melanin produced through α-MSH treatment by 46.54%, confirming that there were no problems with this experiment.

[0122] In samples containing 2 parts by weight or less of ginger extract as an active ingredient, the 0.5 part by weight treatment group showed a statistically significant level of inhibition of extracellular melanin production (p<0.05), and a 0.5% concentration demonstrated a 16.20% inhibitory effect on extracellular melanin production.

[0123] [Table 3]

[0124] [Table 4]

[0125] (3) Effect of the sample on intracellular melanin production

[0126] The effects of β-arbutin, a positive control group, and the sample on melanin production in B16-F10 melanoma cells were examined.

[0127] Figure 4 is a graph showing the amount of intracellular melanin released from the sample, and Figure 5 is a graph showing the inhibition rate of intracellular melanin production from the sample.

[0128] Table 5 below shows the test results for the amount of melanin produced intracellularly in the sample, and Table 6 below shows the test results for the inhibition rate of melanin production intracellularly in the sample.

[0129] As shown in Tables 5 and 6 below, in the case of β-arbutin, the positive control group, it was confirmed that intracellular melanin produced through α-MSH treatment was inhibited by 36.08%, confirming that there were no problems with this experiment.

[0130] In the case of the sample, intracellular melanin production was confirmed to be inhibited at a statistically significant level (p<0.05) in the concentration treatment groups of 0.125 parts by weight to 0.5 parts by weight, and a concentration of 0.5 parts by weight showed a 28.76% inhibitory effect on intracellular melanin production.

[0131] [Table 5]

[0132] [Table 6]

[0133] (4) The effect of the sample on total melanin production

[0134] [Table 7]

[0135] [Table 8]

[0136] Figure 6 is a graph showing the total amount of melanin released from the sample, and Figure 7 is a graph showing the total melanin production inhibition rate of the sample.

[0137] Table 7 shows the results of the total melanin production test for the samples, and Table 8 shows the results of the total melanin production inhibition rate test for the samples.

[0138] The effects of β-arbutin, a positive control group, and the sample on the total amount of melanin produced inside and outside B16-F10 melanoma cells were examined.

[0139] As shown in Figures 6 and 7, and Tables 7 and 8, β-arbutin, the positive control group, was found to inhibit total intracellular and extracellular melanin production by 38.47% through α-MSH treatment.

[0140] In the case of the sample, it was confirmed that total intracellular and extracellular melanin production was inhibited at statistically significant levels (p<0.05) at treatment concentrations of 0.125% to 0.5%, and it was shown that total intracellular and extracellular melanin production was inhibited by 25.88% at a concentration of 0.5%.

[0141] K. Experimental results

[0142] In this study, we were able to confirm the effects on total melanin production by examining the effects of the sample on cell viability in B16-F10 melanoma cells, the effects on melanin produced within cells, and the effects on melanin secreted outside cells.

[0143] The results of examining the effect of the sample on the viability of B16-F10 melanoma cells showed that, at treatment concentrations of 2 parts by weight or less, the cell viability was over 90% compared to the untreated group, confirming that the sample did not affect the cells.

[0144] The results of our investigation into the effects of β-arbutin on melanin production in B16-F10 melanoma cells showed that it inhibited total intracellular and extracellular melanin production by 38.47% in the positive control group, confirming that this experiment was not abnormal.

[0145] At treatment concentrations of 0.125 parts by weight to 0.5 parts by weight of the sample, the treatment was found to inhibit total intracellular and extracellular melanin production at a statistically significant level (p<0.05). In particular, at a concentration of 0.5 parts by weight, it was confirmed that total intracellular and extracellular melanin production was inhibited by 25.88%.

[0146] In conclusion, the sample was confirmed to inhibit total melanin production both inside and outside cells, and is therefore judged to have skin-whitening efficacy.

[0147] Example 4. Efficacy test for collagen production promotion of a composition containing ginger extract as an active ingredient.

[0148] In Example 4, a test was conducted to confirm the collagen production promoting efficacy of a sample (hereinafter referred to as "the sample") containing 4 parts by weight or less of ginger extract extracted using methylene chloride as a solvent as an active ingredient.

[0149] Collagen, present in human skin, is a major structural protein of the extracellular matrix (ECM) of the skin, responsible for skin elasticity and supporting skin structure.

[0150] This type of collagen is synthesized as procollagen within cells, and then secreted outside the cell where it can polymerize into collagen fibers.

[0151] The extracellular matrix (ECM) is a macromolecular network that fills the extracellular space of the dermis, providing the structure and force that supports dermal tissue. Changes in the shape and structure of this ECM tissue require degradation of the ECM, and ECM degradation and resynthesis are also required during tissue repair after inflammation and tissue damage.

[0152] Therefore, regardless of the progression of aging, ECM collagen is constantly being broken down and newly synthesized for tissue reconstruction, a process known as collagen homeostasis.

[0153] However, while collagen synthesis decreases due to external factors such as aging and UV exposure, collagen degradation increases. This breakdown of collagen homeostasis is known to be closely linked to the breakdown of the extracellular matrix (ECM) structure and the resulting wrinkle formation in the skin.

[0154] This study aims to confirm the amount of procollagen released from cells into the cell culture medium and to verify the anti-aging activity of the test substance.

[0155] In this study, the effects of the sample on collagen biosynthesis in dermal fibroblasts were investigated in vitro.

[0156] A. Cell culture

[0157] Human-derived fibroblast cell line CCD-986sk cells are cultured in IMDM medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin in a 150 mm cell culture dish in a 5% CO2 incubator. Cells that reach confluence are maintained through subculturing using mild trypsin.

[0158] I. Cell viability experiment

[0159] Dehydrogenases present in mitochondria of living cells produce a chromogenic substance called formazan using tetrazolium salt (WST).

[0160] Therefore, by measuring this, the number of living cells can be determined. 3 × 10⁶ cells in a 96-well cell culture plate. 3 CCD-986sk cells at a cell / well concentration are cultured for 24 hours in IMDM culture medium supplemented with 10% FBS and 1% P / S.

[0161] After 24 hours, the samples are starved for at least 6 hours in FBS-free IMDM supplemented with 1% P / S, and then treated with IMDM (FBS-free, 1% P / S) medium. The samples are dissolved in DW, and stocks are prepared by diluting them to different concentrations. These stocks are then diluted in IMDM (FBS-free, 1% P / S) medium to the final concentration and incubated alternately for 48 hours.

[0162] To change the culture medium, suction out all of the previous culture medium and add the newly prepared medium. After adding IMDM (FBS-free, P / S 1%) medium containing 10% EZ-CYTOX to the cultured cells and allowing them to react for 2 hours, the cell viability is determined by measuring the absorbance at 450 nm using a plate multireader.

[0163] Cell viability (%) = (Absorbance of sample-treated group / Absorbance of untreated group) × 100

[0164] U. Type I procollagen expression-promoting EIA assay

[0165] Enzyme-linked immunosorbent assay (ELISA) was performed using the Procollagen Type I C-peptide (PIP) EIA Kit (TAKARA MK101) according to the protocol provided by the manufacturer.

[0166] Dispense 100 μL / well of antibody-POD conjugate solution into each well strip provided in the kit, mix 20 μL / well of standard solution and sample, and then incubate at 37°C for 3 hours.

[0167] After removing the supernatant, wash the wells four times with wash buffer. Add 100 μL of Substrate Solution (TMBZ) to each well and let stand at room temperature (20-30°C) for 15 minutes.

[0168] The reaction is stopped by adding 100 μL / well of stop solution, and the absorbance at 450 nm is measured using a multiplate reader.

[0169] The total protein content of each sample was quantified using a BCA assay, and the type I procollagen expression levels of the untreated group and the sample-treated group, corrected for total protein content, were compared to calculate the type I procollagen expression enhancement rate.

[0170] Type I procollagen expression enhancement rate (%) = (Expression level of the corrected sample-treated group / Expression level of the corrected untreated group) × 100

[0171] Protein content was confirmed using the Pierce BCA Protein Assay Kit (23227, Thermo Scientific). 200 μL of Pierce BCA Protein Assay Reagent mixture was added to 20 μL of culture medium and mixed. After standing at 37°C for 30 minutes, the absorbance was measured at 570 nm. Albumin Standard (23209, Thermo) was used as a standard substance to confirm protein content.

[0172] E. Data analysis and statistical processing

[0173] The statistical significance of all experimental results was confirmed using the t-test (independent t-test, two-tailed test, unequal variance assumption) function in Microsoft Office Excel 2007, and a p-value of less than 0.05 was considered statistically significant.

[0174] O. Effects on human-derived fibroblast survival rate

[0175] Table 9 below shows the results of the cytotoxicity test on the samples, and Figure 8 is a graph showing the cell viability of the samples.

[0176] The effects of the samples on the viability of the human fibroblast cell line CCD-986sk were examined and are shown in Table 9. As a result, the "Ginger6 ginger water" sample showed a cell viability of over 90% at concentrations of 4% or less.

[0177] [Table 9]

[0178] C. Effects on type I procollagen expression

[0179] Table 10 shows the results of the type I procollagen expression level measurement test for the samples, and Figure 9 is a graph showing the type I procollagen expression levels of the samples.

[0180] The results of examining the effect of the sample on type I procollagen production in human derived fibroblasts are shown in Table 10 below. The results showed that the sample exhibited a concentration-dependent effect in the range of 0.125 to 0.5 parts by weight, and type I procollagen expression was statistically significant (p<0.05).

[0181] In particular, it was confirmed that treating the sample with a concentration of 0.5 parts by weight promoted type I procollagen expression by approximately 48.01% compared to the untreated control group.

[0182] When TGF-β1, used as a positive control group, was treated at a concentration of 10 ng / mL, it showed a 77.52% increase in type I procollagen expression compared to the untreated control group, confirming that there were no problems with the experiment.

[0183] [Table 10]

[0184] K. Test results

[0185] This study aims to confirm the effect of the sample on the production of type I procollagen in human derived fibroblasts, thereby confirming its potential as a wrinkle-improving and anti-aging cosmetic ingredient in vitro.

[0186] The results of examining the effect of the sample on the viability of human derived fibroblasts showed that at a treatment concentration of 4 parts by weight or less, the cell viability was over 90% compared to the untreated control group, confirming that it did not affect the viability of CCD-986sk cells.

[0187] The results of our investigation into the effect of CCD-986sk, a human derived fibroblast, on type I procollagen production showed that, in the case of TGF-β1 used as the positive control group, treatment at a concentration of 10 ng / mL increased procollagen production by 77.52% compared to the untreated control group, confirming that this experiment was not abnormal.

[0188] The samples showed concentration-dependent reactions in the range of 0.125 to 0.5 parts by weight, and demonstrated statistically significant levels (p<0.05) of type I procollagen production. In particular, when the samples were treated with a concentration of 0.5 parts by weight, type I procollagen expression increased by 48.01% compared to the untreated control group.

[0189] In conclusion, the sample was determined to possess type I procollagen production-promoting activity in a concentration range of 0.125 to 0.5 parts by weight, and is therefore deemed usable as an anti-aging material.

[0190] The scope of protection of the present invention is not limited to the description and expression of the embodiment explicitly stated above. Furthermore, it is reiterated that the scope of protection of the present invention is not limited by obvious modifications or substitutions in the art to which the present invention pertains.

Claims

1. It contains ginger extract as an active ingredient, The ginger extract is extracted using methylene chloride. A composition containing ginger extract as an active ingredient, which has skin whitening and skin elasticity improving effects.

2. The ginger extract is, Methylene chloride extraction is carried out at 10°C for 50 hours, followed by extraction at 25°C for 30 hours. A composition having skin whitening and skin elasticity improving effects, comprising the ginger extract described in claim 1 as an active ingredient.

3. The ginger extract is, It is contained in an amount of 0.125 to 0.5 parts by weight per 100 parts by weight of the total. A composition having skin whitening and skin elasticity improving effects, comprising the ginger extract described in claim 1 as an active ingredient.

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