Uses of safflower extract for improving skin condition

Abstract

To provide the use of Crassocephalum rabens extract in improving skin conditions.SOLUTION: Crassocephalum rabens extract disclosed herein has activity in enhancing expression of collagen gene and / or of elastin gene. By administrating an effective amount of the Crassocephalum rabens extract or a composition containing the effective amount of the Crassocephalum rabens extract to an individual, the levels of collagen and elastin in the individual's skin can be effectively increased to improve skin texture and retard skin aging.SELECTED DRAWING: Figure 13B

Description

Technical Field

[0001] The present invention relates to a second use of plant extracts, and particularly to the use of safflower centaury extracts for improving skin conditions.

Background Art

[0002] Modern people are increasingly emphasizing appearance, and particularly care about the quality of their skin. There are many factors that affect the quality of the skin, including not only an individual's genes, but also diet, lifestyle, exercise habits, environment, pollution, and age. For example, aging reduces collagen and elastic fibers, making the skin appear less elastic and shiny, and causing wrinkles. Ultraviolet rays generate a lot of reactive oxygen species in the body, causing skin spots and wrinkles.

[0003] Safflower centaury is a herbaceous plant of the Asteraceae family and is also called fairy vegetable in Taiwan. The Pharmacopoeia of the People's Republic of China states that safflower centaury has the effects of clearing heat and detoxifying, promoting qi movement and diuresis, and is used for the treatment of hypertension, headache, and insect bites. In recent research, it has been shown that certain active ingredients contained in safflower centaury can treat cancer and suppress inflammatory reactions, and thus can be used as an anticancer drug or an anti-inflammatory drug. Also, research has proven that the active ingredient dLGG (1,2-di-O-linolenoyl-3-O-β-galactopyranosyl-sn-glycerol) in safflower centaury can reduce the recurrence of breast cancer and suppress lung metastasis. [Summary of the Invention] [Problems to be Solved by the Invention]

[0004] The main object of the present invention is to provide a use of safflower centaury extract for improving skin conditions. That is, by administering an effective amount of the safflower centaury extract disclosed in the present invention to an individual, the contents of collagen and elastin in skin cells can be increased, thereby achieving the effects of maintaining skin turgidity, reducing wrinkle formation, and improving skin grooves. [[ID=2�]]

[0005] The main objective of the present invention is to provide a use for safflower extract in improving skin condition. The safflower extract disclosed in the present invention has anti-hyaluronidase activity, inhibitory activity against inflammatory factors and ROS production, and antioxidant activity. Therefore, by administering an effective amount of the safflower extract disclosed in the present invention to an individual, it is possible to improve skin water content, improve skin blemishes, and reduce the occurrence of skin hypersensitivity. [Means for solving the problem]

[0006] To achieve the above objectives, the present invention provides uses for compositions containing safflower extract for improving skin texture or skincare. Specifically, compositions containing safflower extract are used to lighten skin blemishes, improve collagen content in skin cells, reduce the number of pores, and / or reduce pore size.

[0007] In one embodiment of the present invention, the safflower extract is obtained from safflower by an extraction process, and the safflower extract contains a predetermined amount of dLGG (1,2-di-O-linolenoyl-3-O-β-galactopyranosyl-sn-glycerol). Here, the extraction process is well known in the present invention and will not be described here.

[0008] In another embodiment of the present invention, the dLGG content in the safflower extract is approximately 200-250 mg / mL.

[0009] In the embodiments of the present invention, the composition containing safflower extract is a nutritional supplement, a food, or a topical skincare product.

[0010] Another embodiment of the present invention provides its use in preparing an anti-wrinkle composition of safflower extract. Specifically, since the safflower extract disclosed in the present invention has the activity to improve the expression of genes related to collagen and / or elastin in the skin, administering the anti-wrinkle composition to an individual can effectively increase the collagen and / or elastin content in the individual's skin, thereby achieving anti-wrinkle or wrinkle-eliminating effects. [Brief explanation of the drawing]

[0011] [Figure 1A] This is an HPLC chromatogram of the safflower extract disclosed in the present invention, where the safflower extract was prepared by supercritical fluid extraction. [Figure 1B] This is an HPLC chromatogram of the safflower extract disclosed in the present invention, where the safflower extract was prepared by an alcohol immersion extraction method. [Figure 1C] This is an HPLC chromatogram of the safflower extract disclosed in the present invention, where the safflower extract was prepared by an extraction method using water immersion. [Figure 2] The figures show the results of HPLC analysis of the safflower extract disclosed in this invention. In the figures, the upper figure is the HPLC chromatogram of the safflower extract disclosed in this invention, and the lower figure is the HPLC chromatogram of the dLGG standard. [Figure 3] This shows the results of measuring the number of pores using the VISIA skin diagnostic device before and after use of the safflower extract disclosed in this invention. [Figure 4A] This is the result of analyzing the change in the number of pores after using the safflower extract disclosed in this invention for different periods of time. [Figure 4B] This is the result of analyzing changes in skin furrows after using the safflower extract disclosed in this invention for different periods of time. [Figure 5] This shows the results of measuring the number of blemishes using the VISIA skin diagnostic device before and after using the safflower extract disclosed in this invention. [Figure 6A] This is the result of analyzing changes in sun-induced lentigines after using the safflower extract disclosed in this invention for different periods of time. [Figure 6B] This is the result of analyzing changes in blemishes after using the safflower extract disclosed in this invention for different periods of time. [Figure 7] The figure shows the results of an analysis of changes in skin wrinkles after using the safflower extract disclosed in this invention, and indicates that the p-value between the two groups is <0.05. [Figure 8] These are the results of measuring wrinkles using the VISIA skin diagnostic device before and after different subjects took the safflower extract disclosed in the present invention. [Figure 9] The figure shows the results of an analysis of the change in collagen content in the skin after using the safflower extract disclosed in this invention, and the p-value between the two groups is <0.001. [Figure 10] These are the results of measuring and analyzing collagen levels in different subjects before and after taking the safflower extract disclosed in this invention. [Figure 11] The figure shows the results of an analysis of changes in skin tone after using the safflower extract disclosed in this invention, and the p-value between the two groups is <0.001. [Figure 12] The figure shows the results of an analysis of changes in the sunspot value of the skin after using the safflower extract disclosed in this invention, and the p-value between the two groups is <0.01. [Figure 13A] This shows the results of measuring the gene expression of COL1A1, COL1A2, ELN, etc., within the cells of each group. [Figure 13B] This is the result of statistical analysis and quantification of Figure 13A. [Modes for carrying out the invention]

[0012] The present invention discloses the use of safflower centaury extract for improving skin condition. Specifically, since the safflower centaury extract disclosed by the present invention has the activity of improving the expression of collagen gene and / or elastin gene, by administering an effective amount of the safflower centaury extract disclosed by the present invention or a composition containing an effective amount of the safflower centaury extract disclosed by the present invention to an individual, the collagen and elastin contents in the skin of the individual can be effectively improved, and effects such as improvement of skin texture and anti-aging of the skin can be achieved.

[0013] In the present invention, "improvement of skin texture" and "improvement of skin condition" refer to reduction in the number of wrinkles, shallowing of skin grooves, improvement of skin color, suppression of freckle formation, narrowing of pores, reduction in the number of pores, fading of existing pigment spots or freckles, improvement of dull skin, improvement of skin flushing, improvement of collagen content, increase in skin swelling degree, increase in skin glossiness, improvement of skin brightness, etc.

[0014] The safflower centaury extract disclosed by the present invention is a mixture rich in dLGG (1,2-di-O-linolenoyl-3-O-β-galactopyranosyl-sn-glycerol) obtained through an extraction process from safflower centaury raw materials. As shown in FIGS. 1A to 1C, the safflower centaury extract disclosed by the present invention can be prepared by various extraction treatments, such as supercritical extraction method, extraction method by alcohol immersion, extraction method by water immersion, or combination of any two of the above methods. No matter which extraction treatment is used to prepare the safflower centaury extract, the main index component thereof is dLGG (1,2-di-O-linolenoyl-3-O-β-galactopyranosyl-sn-glycerol).

[0015] The term "safflower centaury raw material" can be a fresh plant body or a powder made from a fresh plant body.

[0016] The term "extraction process" includes methods of separating specific components from a mixture using various solvents, including extraction methods by water immersion, extraction methods by alcohol immersion, supercritical extraction methods, or any combination of the above-mentioned extraction methods. Here, the extraction solvent used in the extraction method by water immersion is water, the extraction solvent used in the extraction method by alcohol immersion is alcohol with a concentration of 95% or more, and the extraction solvents in the supercritical extraction method are carbon dioxide and alcohol with a concentration of 95% or more.

[0017] The term "composition" refers to a product that can be used to improve or maintain skin quality, the usage method is not limited, including external use, internal use, etc., and its dosage form is also not limited, for example, emulsion, cream, liquid, tablet, powder, capsule, etc. As an example, the composition disclosed in the present invention can be an external skin care product, a nutritional supplement, a food, etc.

[0018] The term "effective amount" refers to the dosage that can achieve the desired effect, and its volume varies depending on parameters such as dosage form, administration method, and the individual to be administered. For example, when the safflower calendula extract disclosed in the present invention is orally administered to an adult and the effect of improving skin quality is to be obtained within 14 days after administration, the effective amount of the safflower calendula extract is 100 - 200 mg / day, preferably 180 mg / day. However, when administered to an adult by the external use method, the effective amount of the safflower calendula extract is about 0.66 mg / day or more, and the preferred dosage is 0.66 - 3.3 mg / day. The external use dosage was obtained by extrapolation from the dosage (10 - 50 ug / mL) used in the cell test.

[0019] The term "administration" refers to the method of providing the safflower calendula extract or its composition disclosed in the present invention to a subject, such as external use or internal use, etc. Among them, external use includes methods such as rubbing and spraying.

[0020] The term "skin spots" refers to visible marks on the skin, for example, those caused by sunburn, pigmented spots, liver spots, solar lentigines, senile lentigines, scars, or other results of pigmentation.

[0021] The term "solar lentigines" refers to spots on the skin caused by excessive melanin production due to ultraviolet radiation, and these spots are usually black or close to black in color.

[0022] The term "melasma" refers to spots caused by pigmentation on or under the skin, which are usually brown or reddish in color and include freckles and other similar spots.

[0023] The term "approximately" refers to the acceptable error range for a particular numerical value, as determined by a person skilled in the art in which this invention pertains. This error range is determined based on the measurement or calculation method of the particular numerical value and is usually within 10% of the particular numerical value.

[0024] The technical features and effects of the present invention will be explained in detail below, with reference to several examples.

[0025] The clinical trials conducted in the following examples all complied with the relevant regulations of the trials, and since the cells used in the cell studies were commercially available cells, deposit was not required.

[0026] Please note that the data in Tables 3 to 5 disclosed in the following examples were obtained by analyzing and comparing images taken with the VISIA skin diagnostic device, and therefore the obtained data does not have a data unit.

[0027] Example 1: Preparation of Safflower Extract

[0028] Fresh safflower powder was taken, and the following steps were carried out in order to obtain a safflower extract: extraction, filtration, centrifugation, hydration, and freeze-drying. In the extraction step, 95% alcohol was used as the extraction solvent. The safflower extract is rich in dLGG (1,2-di-O-linolenoyl-3-O-β-galactopyranosyl-sn-glycerol), which was used in the following examples.

[0029] The results of HPLC analysis of the safflower extract disclosed in this invention are shown in Figure 2. The HPLC conditions included a solvent of 98% methanol, a flow rate of 1.0 mL / min, a gradient elution method, an injection volume of 10 μL, a stop time of 60 minutes, a column temperature of 30°C, and detection at a wavelength of 210 nm. From the results in Figure 2, it can be seen that dLGG was detectable in the safflower extract disclosed in this invention at a retention time of 13.05 ± 0.80 minutes, and the dLGG content was 218.05 mg / mL.

[0030] Example 2: Skin Improvement Efficacy Test (1)

[0031] A total of 12 subjects were recruited, and each was given a capsule containing 180 mg of safflower extract. The dosage was one capsule daily, and the study period was 4 weeks. The skin condition of each subject was examined in weeks 0, 2, and 4 of the study, and self-comparisons were performed before and after the study. The VISIA skin diagnostic device was used to examine the skin condition.

[0032] The results in Figures 3 and 4 show that nearly 92% of subjects experienced improvement in skin pores and furrows, with improvements ranging from 7% to 11%. Among these subjects, the most significant improvement was seen in those whose pores improved by approximately 49% after use.

[0033] The results shown in Figures 5 and 6 indicate that nearly 70% of the subjects experienced improvement in their blemishes, demonstrating that the safflower extract disclosed in this invention can prevent the formation of blemishes caused by melanin pigment deposition. Among these subjects, the one who showed the most significant improvement had approximately 28% improvement in both blemishes and pores after use.

[0034] Example 3: Skin Improvement Efficacy Test (II)

[0035] A total of 20 subjects were recruited and randomly divided into two groups for a double-blind study. One group was designated as the test group and given capsules containing 180 mg of safflower extract, with a daily dose of one capsule. The other group was the placebo group. The study period was 4 weeks, and the skin condition of each subject was examined at weeks 0, 2, and 4 of the study. The items examined were skin tone, moisture, elasticity, blemishes, sunspots, melasma, redness, wrinkles, skin furrows, pores, protoporphyrin, and collagen density. The results are shown in Figures 5 to 10.

[0036] The results in Figures 7 and 8 show that after the trial was conducted and the subjects used the product, wrinkles improved by more than 50%, and compared to the placebo group, the improvement in the test group reached 32.7%. This means that the safflower extract disclosed in this invention can effectively reduce the number of wrinkles.

[0037] The results in Figures 9 and 10 show that after the test was conducted and the subjects used the product, the collagen content in the skin increased, with the collagen content in the test group increasing by 8.2% compared to the placebo group. From these results, it can be seen that the safflower extract disclosed in this invention can effectively improve the collagen content in the skin, maintaining elasticity and luster in the skin.

[0038] The results shown in Figures 11 and 12 indicate that, compared to the placebo group, the subjects in the test group showed increased skin glossiness and a reduction in UV-induced blemishes. This demonstrates that the safflower extract disclosed in this invention can effectively improve skin glossiness, maintain skin tone uniformity, and suppress pigmentation.

[0039] Example 4: Comparison of ingredients with different effects

[0040] The safflower extract disclosed in this invention and other commercially available raw materials were prepared, and hyaluronic acid (taken for 20 days), ceramide, and collagen (taken for 6 weeks) were administered orally to compare skin moisture retention, anti-wrinkle effects, and skin collagen density. The results are shown in Table 1 below.

[0041] JPEG0007837058000001.jpg78165

[0042] The results in Table 1 show that, compared to other commercially available ingredients, the safflower extract disclosed in this invention can significantly reduce the occurrence of wrinkles in the skin, eliminate or lighten existing wrinkles, and furthermore, it can effectively stimulate collagen production in the skin, increasing the collagen density in the skin, thereby achieving improved skin quality and anti-aging effects.

[0043] Example 5: Skin Improvement Efficacy Test (III)

[0044] In this study, 40 subjects were recruited and randomly assigned to either a placebo group or a treatment group. As shown in Figure 2, the male-to-female ratio was the same in both groups. The mean ages of the placebo group and the treatment group were 46.4 years and 43.9 years, respectively, and there was no statistically significant difference in the mean ages between the groups. The study period was a total of 4 weeks, and the treatment group was given capsules containing 180 mg of safflower extract, taken at a dose of one capsule daily.

[0045] JPEG0007837058000002.jpg28155

[0046] After the trial, skin changes, including pores, elasticity, wrinkles (skin furrows), and collagen content, were compared between the placebo group and the treatment group. The results are shown in Table 3.

[0047] Table 3 shows that the treatment group had significantly better skin elasticity than the placebo group at week 4. Regarding changes in skin furrows, the placebo group showed a 19.2% increase, while the treatment group showed a 12.8% decrease. Furthermore, subjects in the treatment group showed a significant reduction in wrinkles around the eyes, decreasing by approximately 3.8% (compared to a 0.1% increase in the placebo group). In terms of collagen content, the treatment group showed a significantly increased collagen content compared to the placebo group.

[0048] The results in Table 3 demonstrate that the safflower extract disclosed in this invention can indeed improve skin elasticity, increase the collagen content of the skin, suppress the formation of wrinkles, and improve existing wrinkles. This means that the safflower extract disclosed in this invention has anti-aging and skin condition improving effects.

[0049] JPEG0007837058000003.jpg161163

[0050] Furthermore, the skin blemishes, sunspots (UV spots), and melasma of the subjects were measured before and after the trial. The results are shown in Table 4.

[0051] Table 4 shows that although there were no significant changes in skin spots and melasma between the two groups, the difference in solar lentigines between week 0 and week 4 in the treatment group was -12.8%, while the mean solar lentigines in the placebo group at week 4 increased by 7.5% compared to the results at week 0.

[0052] The results in Table 4 show that the safflower extract disclosed in this invention can suppress the formation of skin spots caused by ultraviolet light. This means that it can effectively prevent the absorption of ultraviolet light by the skin and reduce the damage that ultraviolet light inflicts on the skin.

[0053] JPEG0007837058000004.jpg102163

[0054] Furthermore, skin brightness, skin moisture content, and red area were measured before and after the trial in both the placebo and treatment groups. The results are shown in Table 5. Of these, the red area indicates that there is a problem of skin redness and / or inflammation.

[0055] The results in Table 5 show that the safflower extract disclosed in this invention significantly improved the brightness of the subjects' skin and enhanced their skin's moisture retention within four weeks of administration. This indicates that the safflower extract disclosed in this invention effectively increases skin luster and prevents the occurrence of skin defects caused by dehydration.

[0056] JPEG0007837058000005.jpg102165

[0057] Example 6: Cell Test

[0058] 3T3 fibroblasts were taken and divided into four groups. Each group was treated with 0, 10, 25, and 50 μg / ml of safflower extract for 48 hours. After completion, the expression of genes such as COL1A1, COL1A2, and ELN in the cells of each group was measured by real-time polymerase chain reaction. The results are shown in Figures 13A and 13B. Of these, the safflower extract contained approximately 7.9% dLGG (1,2-di-O-linolenoyl-3-O-β-galactopyranosyl-sn-glycerol), and the volume of each culture well was 5 ml. That is, 50 to 250 μg of safflower extract was added to each culture well.

[0059] The results in Figures 13A and 13B show that, compared to cells not treated with the safflower extract disclosed in this invention, the expression of genes such as COL1A1, COL1A2, and ELN in cells treated with the safflower extract disclosed in this invention was increased. Of these, the expression levels of COL1A1 in the cells were almost the same in each group, but for genes such as COL1A2 and ELN, the group treated with 10 μg / ml of safflower extract showed superior results.

[0060] As is well known to those skilled in the art, the expression of COL1A1 and COL1A2 is related to collagen proliferation, and ELN is related to elastin production. In other words, an increase in the expression of COL1A1, COL1A2, and ELN indicates an increase in the collagen and elastin content in the skin. Therefore, the results of this example show that the safflower extract disclosed in this invention has the activity to increase the expression of genes related to skin structure, such as COL1A1, COL1A2, and ELN, in cells, thereby achieving the effect of maintaining skin luster and elasticity and improving skin wrinkles.

[0061] Furthermore, if the standard for dLGG to be contained in a general topical composition is approximately 0.6%, then, according to the results of Example 6, the topical dose of safflower extract would be at least approximately 0.66 mg / day, and the preferred topical dose is 0.66 to 3.3 mg / day.

[0062] The results of the above-mentioned examples demonstrate that the safflower extract disclosed in this invention has no side effects on individuals and, through inhibitory mechanisms such as anti-hyaluronidase activity, inflammatory factor and ROS generation, and antioxidant activity, it achieves improvement in skin water content and suppression of collagen and elastin degradation. This effectively improves the elasticity of the subject's skin, reduces wrinkles, increases collagen content in the skin, reduces skin hypersensitivity, improves skin brightness, and improves blemishes.

Claims

1. A composition for use in improving skin condition or skincare, wherein the composition is used to lighten skin blemishes, increase the collagen content in skin cells, increase the elastin content in skin cells, reduce the number of pores, or reduce the size of pores, and contains safflower extract, the safflower extract is obtained by carrying out the following steps in order from safflower powder: extraction, filtration, centrifugation, hydration, and freeze-drying, and the safflower extract is rich in dLGG (1,2-di-O-linolenoyl-3-O-β-galactopyranosyl-sn-glycerol).

2. The composition according to claim 1, which is a nutritional supplement.

3. The composition according to claim 1, which is a food product.

4. The composition according to claim 1, which is an external skincare product.

5. A composition comprising safflower extract for use in the preparation of an anti-wrinkle composition.

6. The composition according to claim 5, which is used to improve the collagen content in skin cells.

7. The composition according to claim 5, which is used to improve the elastin content in skin cells.

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