AU2012286357A1 - Eucommia extract, preparation method therefor and use thereof - Google Patents

Abstract

A preparation method of a eucommia extract comprises: (1) solvent extraction, wherein the solvent is selected from water, methanol, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone, ethyl acetate, or a combination thereof; (2) separation of the extract and residue, to obtain an extract; (3) optional recovery of the solvent; and (4) optional dissolution and dilution. The resulting eucommia extract has antioxidant and is anti-UVA and UVB radiation. The resulting eucommia extract can be used as an active ingredient to prepare anti-skin aging, especially anti-skin photoaging cosmetics or medicines.

Description

EUCOMMIA EXTRACT, PREPARATION METHOD THEREFOR AND USE THEREOF Field of the invention The present invention relates to natural pharmaceutical chemistry field and cosmetic field, particularly to a Chinese herbal medicine Eucommia ulmoides for anti-aging, especially for anti-photoaging for use in a cosmetic or a medicament. Background of the invention Studies have indicated that free radicals and UV are one of the factors of skin aging, and that the photo-aging is mainly caused by both the medium-wavelength ultraviolet (UVB, 290~320 nm) together with the long-wavelength ultraviolet (UVA, 320~400 nm) in the sunlight. Ultraviolet rays significantly hasten aging of the human skin, with one-fold higher risk of skin aging and 10-year earlier aging occurrence in high-exposure population as compared with low-exposure population. The long-term exposure to the ultraviolet rays of sunlight results in the light aging of the skin and can lead to serious cosmetic issues, therefore more and more professional attentions are drawn in recent years. Anti-aging and anti-photoaging products have been released in succession; and more related researches are being conducted. However, anti-photoaging additives of Chinese herbal medicine remained vacant. Description of the invention The purpose of the present invention is to provide a method for preparing an extract of Eucommia ulmoides, comprising: (1) extracting with a solvent, wherein the solvent is selected from water, methanol, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone, ethyl acetate, or a combination thereof; (2) separating an extract and residue to obtain the extract; (3) optionally, recovering the solvent; and (4) optionally, dissolving or diluting. Preferably, the extraction of step (1) is selected from soaking, decoction, refluxing, 1 diacolation or a combination thereof. Preferably, the solvent used in step (4) is selected from water, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone, ethyl acetate or a combination thereof. Another purpose of the present invention is to provide another method for preparing an extract of Eucommia ulmoides, comprising: (1) extracting with a solvent, wherein the solvent is selected from methanol, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone, ethyl acetate, or a combination thereof; (2) separating an extract and residue to obtain the residue; (3) extracting the residue of step (2) with water; (4) separating an water extract and residue to obtain the water extract; (5) concentrating, precipitating with ethanol; and separating to obtain the precipitate; and (6) optionally, dissolving and formulating the precipitate of step (5) into a solution. Preferably, the extraction of steps (1) and (3) is selected from soaking, decoction, refluxing, diacolation or a combination thereof. Preferably, the solvent used in step (6) is selected from ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone, ethyl acetate or a combination thereof. Yet another purpose of the present invention is to provide the extract of Eucommia ulmoides obtained according to the above preparation method of the present invention. Preferably, the extract of Eucommia ulmoides is in a form selected from emulsion, solution, powder, ointment, film or cream. The extract of Eucommia ulmoides obtained according to the method of the present 2 invention can be used directly to prevent the skin from aging, especially from light aging, or can be added to the cosmetics or medicaments as functional additives or active ingredients. Yet another purpose of the present invention is to provide a use of the extract of Eucommia ulmoides in the preparation of anti-aging, especially anti-photoaging cosmetics or medicaments. Preferably, the extract of Eucommia ulmoides is the extract of Eucommia ulmoides obtained according to the method of the present invention. Yet another purpose of the present invention is to provide an anti-aging, especially anti-photoaging cosmetics or medicaments comprising the extract of Eucommia ulmoides. Preferably, the extract of Eucommia ulmoides is the extract of Eucommia ulmoides obtained according to the method of the present invention. In the present application, Eucommia ulmoides refers to bark of Eucommiaceae plant Eucommia ulmoides oliv. Eucommia ulmoides oliv is mainly produced in Sichuan, Shaanxi, Hubei, Henan, Guizhou and Yunnan. During Qingming to the summer solstice, the plants grown between 15-20 years are selected, the bark thereof is taken, of which the rough cortex is removed, dried and placed in ventilated and dry place according to the size and specification of the medicine. Eucommia ulmoides has sweet, acrid taste, is warm in property and can tonify liver and kidney, strengthen the bones and muscles, prolong life and beautify the face through the liver and kidney. Eucommia ulmoides contains organic compounds such as lignans, iridoids, organic acids, terpenoids, polysaccharides, amino acids, gutta percha and the like, and inorganic elements such as calcium, iron and the like. "BIE LU" recorded that "Do not want to practice because of the pain in foot", "Shen Nong's Herbal Classic" recorded that "it can tonify the spirit, strengthen the bones and muscles, and make the body relaxed and healthy after a long time administration". Modem pharmacological studies show that Eucommia ulmoides has anti-aging 3 effects, can resist lipid peroxidation and has an effect of adjusting the immune function by a two-way. The extract of Eucommia ulmoides obtained according to the method of the present invention has anti-aging effect by scavenging free radicals, and can help the skin against the ultraviolet radiation of UVB and UVA, protect the skin, reduce the damage of cell, prevent the reduction of the dermal collagen, effectively prevent the skin from aging, especially light aging, without any irritation on the skin. Embodiments Example 1 For 100g Eucommia ulmoides , 8-fold amount (fold amount refers to the ratio between the volume of the solvent (ml) and the weight of Eucommia ulmoides (g), hereinafter the same) of 75% (v/v) ethanol was added. The mixture was kept cold soaking for 48 hours and then filtered. Ethanol was recovered from filtrate; and the resultant extracts were dissolved with water to make a solution with a concentration of crude drug of 1000 mg/ml, thus obtaining extracts A in the form of solution. Example 2 For 100g Eucommia ulmoides , 8-fold amount of methanol was added. The mixture was subjected to refluxing extraction for 2 hours, and filtered. Methanol was recovered from filtrate; and the resultant extracts were dissolved with water to make a solution with a concentration of crude drug of 500 mg /ml, thus obtaining extracts B in the form of solution. Example 3 100g Eucommia ulmoides was decocted with 10-fold amount of water for about 2 hours, concentrated and filtered. The supernatant was made into a solution with a concentration of crude drug of 1000 mg /ml by adding water, thus obtaining extracts C in the form of solution. 4 Example 4 For lOOg Eucommia ulmoides, 6-fold amount of ethanol was added. The mixture was subjected to refluxing extraction for 2 hours, and filtered. The residue was decocted with 10-fold amount of water for about 2 hours, concentrated, precipitated with 2-fold amount of ethanol, and filtered. The precipitates were dissolved with water to make a solution with a concentration of crude drug of 500 mg /ml, thus obtaining extracts D in the form of solution. Example 5: DPPH experiment Free radical is a class of chemical substance with active reactivity and extremely high oxizability. The radiation of ultraviolet UVA/UVB can generate numerous free radicals which damage the body and accelerate aging. DPPH free radical scavenging assay was used herein to determine the in vitro anti-oxidation activity of the extracts of the examples. The extracts of the examples were diluted with distilled water to four concentrations, 5%, 2.5%, 1.25% and 0.625% (v/v), respectively. Two milliliters of each of the diluted extracts of the Examples, 0.5 ml of DPPH solution in ethanol at a concentration of 500 [tmol/L, and 1.5ml water were then sequentially added into a same test tube and shaken thoroughly. After being left stand for 30 min, its optical density (GD) was measured at 517 nm by using a corresponding mixture liquid as the blank control. The scavenging rates for DPPH of the extracts of the examples were calculated according to the following formula: The scavenging rate (%) - T 100 % wherein: T -- the optical density of DPPH + the extracts of the Examples, TO -- the optical density of the extracts of the examples + solvents, C -- the optical density of DPPH + solvents. The results were shown in Table 1. 5 Table 1. Evaluation of the oxidation resistance of the extract of Eucommia ulmoides Free Free radical Free radical Free Concen- radical Concen- Concen- Concen- radical scavenging scavenging trt saegn tration scavenging tration tration traction scavenging rate rate rate rate Extracts A of 5% 63.28% 2.5% 45.98% 1.25% 33.88% 0.625% 23.02% Example 1 Extracts B of 5% 69.88% 2.5% 49.89% 1.25% 38.76% 0.625% 25.04% Example 2 Extracts C of 5% 70.38% 2.5% 51.00% 1.25% 39.60% 0.625% 27.56% Example 3 Extracts D of 5% 74.12% 2.5% 56.49% 1.25% 44.96% 0.625% 32.67% Example 4 1 1 1 The results of the experiments showed that, the extracts A of Example 1, the extracts B of Example 2, the extracts C of Example 3 and the extracts D of Example 4 had superior antioxidant ability for free radical scavenging in vitro at all four concentrations, wherein the effect from the extracts D of Example 4 was the best. Example 6: proliferation experiment of fibroblast The fibroblast in the skin is the main cells for collagen synthesis. The large proliferation of fibroblast, or the increased ability of fibroblast for synthesizing and secreting collagen can all increase the total amounts of collagen, thus postponing the skin aging. Hence, studying the effect of promoting proliferation of fibroblast is one of the important methods to evaluate the anti-aging activity of the sample. The present experiment uses the method of culturing human fibroblast in vitro to preliminarily evaluate the anti-aging efficacy of the extract of Eucommia ulmoides according to the present invention based on the above skin aging mechanism. The prepared cell suspension (formulated into a concentration of 5*10 3 /ml) was seeded into a 96-well plate. Except the blank control, 20 1 1 Chinese herbal medicine 6 extracts of Examples with corresponding concentrations was added to each well after the cell attachment. Four concentration groups were set with 6 wells per group. The plate was placed into an incubation chamber (37 'C and 5% C0 2 ) to incubate for 24 hours, then 20 yi 1 MTT solution was added. The plate was continued to incubate in the incubation chamber (37 'C and 5% C0 2 ) for 4 hours. The enzyme panel was taken out, the culture solution in each well was absorbed carefully, and the DMSO was added into the wells and mixed evenly. The optical density (570 nm) was measured by an enzyme-linked immunosorbent assay apparatus and the relative proliferation rate was obtained by comparison with the control. The results of proliferation activity of the extract of Eucommia ulmoides to the fibroblast were shown in Table 2. Table 2: proliferation activity of the extract of Eucommia ulmoides to the fibroblast The concentrations of the Chinese herbal medicine 1% 0.1% 0.01% 0.001 composition according to the % present invention The Extracts A of 18.86 14.60 10.86 9.22% proliferation Example 1 rate for Extracts B of 18.92 14.44 11.36 9.88% promoting Example 2 % % % Extracts C of 22.26 20.98 16.24 11.91 dermal Example 3 % % % % fibroblast Extracts D of 20.04 18.78 13.86 10.02 Example 4 % % % % The results of the experiments indicated that extracts A of Example 1, extracts B of Example 2, extracts C of Example 3 and extracts D of Example 4 all could promote the proliferation of the fibroblast and the effect from the extracts C of Example 3 was the best, wherein the proliferation rate of fibroblast was promoted to be 11.91% at a concentration of 0.001%. Example 7: Protective effect on the fibroblast against the damage of UV radiation 7 Light aging of the skin is principally caused by the medium-wavelength ultraviolet rays (UVB, 290~320 nm) together with the long-wavelength ultraviolet rays (UVA, 320~400 nm) in the sunlight, and the ultraviolet rays significantly hasten aging of human skin. UVA/UVB radiation will result in damage and death of fibroblasts and hurt the skin, leading to light aging. The anti-light aging effect of the Chinese herb medicine extracts of the Examples on protecting the cells against damage from UV radiation was investigated herein by UV radiation assay. PBS (4.5 ml) was added to 0.5 ml of each extract of the Examples, to formulate a dilution with a concentration of 1 / 10 (v/v), which was subjected to suction filtration. Then 0.1 ml filtrate was added into 9.9 ml sterile PBS and diluted, finally obtaining a dilution with a concentration of 1/1000 (v/v). Skin fibroblasts (Fbs) cultured with a normal DMEM solution were used as the control group, Fbs cultured with a normal DMEM solution, after being irradiated with UVA (5-20J/cm 2 ) / UVB (20-80J/cm 2 ), were used as the model group, and Fbs co-cultured using the extracts of the Examples (at a concentration of 1/1000) along with DMEM solution, after being irradiated with UVA/UVB, were used as the extracts-treated group. After ultraviolet treatment for 24 hours, the optical density (OD value) of each well was measured using a microplate reader with CCK-8. The cell proliferation rate was calculated as the percentage of the OD value of each treated group relative to the OD value of negative control group; each test was run in triplicate. The results of the experiments were shown in Table 3 and Table 4. Table 3. UVA Radiation Cell proliferation rate P value Extracts A of Example 1 (90.13±4.38)% <0.01 Extracts B of Example 2 (90.25±4.20)% <0.01 Extracts C of Example 3 (90.46±3.64)% <0.01 Extracts D of Example 4 (99.77±6.59)% <0.01 Control group (100±2.66)% Model group (78.25±3.98)% <0.01 8 Table 4. UVB Radiation Cell proliferation rate P value Extracts A of Example 1 (97.88±3.53)% <0.01 Extracts B of Example 2 (98.06±2.55)% <0.01 Extracts C of Example 3 (98.19±5.58)% <0.01 Extracts D of Example 4 (99.33±4.60)% <0.01 Control group (100±5.68)% Model group (83.63±5.15)% <0.01 The results of the experiments showed that, all of the extracts A of Example 1, extracts B of Example 2, extracts C of Example 3 and extracts D of Example 4 could reduce the damage of UV radiation to fibroblasts and allowed the cell proliferation rate to be recovered above 90%, wherein the effect from the extracts D of Example 4 was the best. Example 8: Effect of UV radiation on the expression of MMP-1 and MMP-3 mRNAs of dermal fibroblast and the Protective effect of Eucommia ulmoides MMP-1 (interstitial collagenase) and MMP-3 (stromelysin) are two main enzymes which degrade the extracellular matrix of the dermal cells. Their abnormal levels of expression will cause imbalance of extracellular matrix synthesis and catabolism in dermal cells, which is an important reason why dermal structure is damaged in the light aging. The dermal fibroblasts were seeded in 60mm culture dishes and incubated with the extracts of the Examples (at a concentration of 1/1000). After UV irradiation 24 hours on the cells, total cell RNAs were extracted and reversely transcribed, and real-time PCR was carried out according to the designed primers. The results of the experiments were shown in Table 5 and Table 6. Table 5 9 MMP-1/p-actin MIMP-3/pj-actin Un-irradiated group 0.2910.04 5.3210.69 UVA irradiated group 0.4010.04' 9.07+1.32 * Extracts A of Example 1 0.3210.02* 6.47+1.66* Extracts B of Example 2 0.3210.03* 6.45_1.18* Extracts C of Example 3 0.31 0.03 * 6.6710.64 * Extracts D of Example 4 0.30 0.03 **1 48+0.92* Compared to un-irradiated group: #P<0.01; and compared to UVA irradiated group: *P<0.05, **P<0.01 Table 6 MMP-1/p-actin MMP-3/p-actin Un-irradiated group 0.2010.03 3.9110.18 UVB irradiated group 0.31+0.03 6.3310.25 Extracts A of Example 1 0.2410.03* 4.6610.28* Extracts B of Example 2 0.24+0.06* 4.65+0.71* Extracts C of Example 3 0.2210.04 * 4.0810.11 ** Extrats D of Example 4 0.2110.03 365+0.52 Compared to un-irradiated group: #P<0.01; and compared to UVB irradiated group: *P<0.05, **P<0.01 The results of the experiments showed that, all of the extracts A of Example 1, extracts B of Example 2, extracts C of Example 3 and extracts D of Example 4 could reduce the abnormal expression of MMP-1 and MMP-3 mRNAs of dermal fibroblast caused by UV radiation, wherein the effect from the extracts D of Example 4 was the best. Example 9: Effect of UV radiation on the expression level of procollagen type I and III of dermal fibroblast and the Protective effect of Eucommia ulmoides The main pathological changes of light aging are in the dermis layer, and dermal fibroblasts are the main cells to maintain the normal structure and function of the dermis. Collagen type I and III are the main components of the extracellular matrix 10 of the dermal cells. Studies have shown that collagen production is reduced in the light aged skin , and procollagen is significantly reduced after 24 hours of UV irradiation. The dermal fibroblasts were seeded in 35mm culture dishes and incubated with the extracts of the Examples (at a concentration of 1/1000). After UV irradiation 24 hours, the supernatant of the cell culture was collected, and the optical density (OD) was measured at 450 nm with a microplate reader by using double antibody sandwich ABC-ELISA method. The levels (ng/ml) of procollagen type I and III in the samples were read on a standard curve according to OD values. Meanwhile the cells were collected, carried out freeze-thaw cycles at -20'C for three times, centrifugated at 4'C at 14000g for 15min, and the quantitative protein in the cell was measured for the supernatant. A 20ul standard substance or sample to be tested was added to 96-well plate, to each well 200ul working fluid BCA was added, and placed at 37'C for 20-30min. The optical density (OD) was measured at 562 nm by a microplate reader, and the protein levels (mg/ml) of the sample to be tested were calculated according to the standard curve. The expression levels of the procollagen type I and III in the cell culture supernatant were corrected by the cell protein level, and expressed as ng/mg. The results of the experiments were shown in Table 7 and Table 8. Table 7 Procollagen type I Procollagen type III (ng/mg) (ng/mg) Un-irradiated group 106.50+5.20 43.07+1.85 UVA irradiated group 64.13+7 21 24 30+2.80' ExtractsA of Example 1 84.37+2.42* 29.77+2.37* Extracts B of Example 2 82.88+1.81* 30.12+2.66* Extracts C of Example 3 83.57+0.91* 30.10+3.65* FIxtracts D ofExample 4 85.60+3.50 ** 31.47+5.45 * Compared to un-irradiated group: #P<0.01; and compared to UVA irradiated group: *P<0.05, **P<0.01 Table 8 11 Procollagen type I Procollagen type III (ng/mg) (ng/mg) Un-irradiated group 123.8015.82 64.8712.65 UTVB irradiated group 74.57+3.89 37.47±2.75 Extracts A of Example 1 94.37A3.84* 40.7712.37* Extracts B of Example 2 92.882.36* 40.1214.29* Extracts C of Example 3 93.5716.73* 40.1012.43* Extracts D of Example 4 114.40L4.77** 44.40±2.80* Compared to un-irradiated group: 4P<0.01; and compared to UVB irradiated group: *P<0.05, **P<0.01 The results of the experiments showed that, UV radiation significantly inhibits the expression of the procollagen type I and III (P<0.01) by fibroblasts. The additions of the extracts of each Example A, B, C and D (at a concentration of 1/1000) could increase the levels of the procollagen type I and III in the cell culture supernatant more or less, wherein the effect from the extracts D of Example 4 was the best. Example 10: Emulsions I, II, III and IV prepared respectively by the extracts A, B, C and D of Eucommia ulmoides obtained by Examples 1-4 The name of raw INCI name % (W/W) materials Emulsion Emulsion Emulsion Emulsion (Supplier) I II III IV Deionized water Water 73.45 73.45 73.45 73.45 Crodamol GTCC Caprylic/capric 6.0 6.0 6.0 6.0 (Croda) triglyceride EHP (Cognis) Ethylhexyl 4.0 4.0 4.0 4.0 palmitate Lanette 0 Cetanol 3.0 3.0 3.0 3.0 (Cognis) Xanthan gum Xanthan gum 0.1 0.1 0.1 0.1 Emumulgin-BA25 Docosyl alcohol 2.5 2.5 2.5 2.5 12 (Cognis) polyether-25 MGS-AV Glyceryl stearate 1.0 1.0 1.0 1.0 (NIKKOL) Sepigel305 Polyacrylamide, 0.8 0.8 0.8 0.8 C13-14 isoparaffin Lauryl alcohol polyether- 1 Butanediol Butanediol 2.0 2.0 2.0 2.0 Pentanediol 1,2-pentanediol 1.0 1.0 1.0 1.0 (Symrise) Glycerol Glycerol 3.0 3.0 3.0 3.0 Liquid Germall Diazolidinyl urea, 0.65 0.65 0.65 0.65 Plus iodopropynyl butylcarbamate, propylene glycol Extracts A of Example 1 2.5 - - Extracts B of Example 2 - 2.5 - Extracts C of Example 3 - - 2.5 Extracts D of Example 4 - - - 2.5 Preparation method: Caprylic/capric triglyceride, ethylhexyl palmitate, cetanol, docosyl alcohol polyether-25 and glyceryl stearate were mixed, heated and dissolved evenly under stirring; Butanediol, 1,2-pentanediol, glycerol, xanthan gum, deionized water were heated and dissolved evenly under stirring; these two portions were mixed evenly under homogeneous conditions; then added Sepigel305. The mixture was cooled down to 50'C and added Liquid Germall Plus and the extracts of Eucommia ulmoides according to the present invention, stirred evenly to obtain the final product. Example 11: Solutions I, II, III and IV prepared respectively by the extracts A, B, C and D of Eucommia ulmoides obtained by Examples 1-4 13 The name of raw INCI name % (W/W) materials Solution I Solution Solution Solution (Supplier) II III IV Deionized water Water 86.33 86.33 86.33 86.33 Butanediol Butanediol 5.0 5.0 5.0 5.0 Propanediol Propanediol 3.0 3.0 3.0 3.0 Pentanediol 1,2-pentanediol 2.0 2.0 2.0 2.0 (Symrise) Liquid Germall Diazolidinyl 0.60 0.60 0.60 0.60 Plus urea, iodopropynyl butylcarbamate, propylene glycol EDTA disodium EDTA disodium 0.05 0.05 0.05 0.05 Sodium Sodium 0.02 0.02 0.02 0.02 Hyaluronate Hyaluronate Extracts A of Example 1 3.0 - - Extracts B of Example 2 - 3.0 Extracts C of Example 3 - - 3.0 Extracts D of Example 4 - - - 3.0 Preparation method: Water, butanediol, propanediol, 1,2-pentanediol, EDTA disodium and sodium hyaluronate were mixed, heated and dissolved evenly under stirring. The mixture was cooled down to 55 0 C and added Liquid Germall Plus and the extracts of Eucommia ulmoides according to the present invention, stirred evenly to obtain the final product. The cosmetic preparations obtained by the formulations of Examples 10 and 11 had a certain effect of anti-oxidation and anti-photoaging. 14 Skin photoaging is mainly caused by both the medium-wavelength ultraviolet (UVB, 290~320 nm) together with the long-wavelength ultraviolet (UVA, 320~400 nm) in the sunlight. Ultraviolet rays significantly hasten aging of human skin. UVA/UVB radiation will result in damage and death of fibroblasts, reduce the dermal collagen and hurt the skin. Incubation of the dermal fibroblasts irradiated by certain strength of UVA/UVB radiation with the extracts of Eucommia ulmoides can effectively reduce the effect of UVA/UVB radiation on the proliferation of the fibroblasts, effectively reduce the changes of the collagen of the dermal fibroblasts, and protect the skin, suggesting that the extracts of Eucommia ulmoides have the protective effect on the fibroblast against the damage of UV radiation and can prevent light aging. 15

Claims (12)

1. A method for preparing an extract of Eucommia ulmoides, comprising: (1) extracting with a solvent, wherein the solvent is selected from water, methanol, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone, ethyl acetate, or a combination thereof; (2) separating an extract and residue to obtain the extract; (3) optionally, recovering the solvent; and (4) optionally, dissolving or diluting.

2. The method according to claim 1, wherein the extraction of step (1) is selected from soaking, decoction, refluxing, diacolation or a combination thereof.

3. The method according to claim 1, wherein the solvent used in step (4) is selected from water, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone, ethyl acetate or a combination thereof.

4. A method for preparing an extract of Eucommia ulmoides, comprising: (1) extracting with a solvent, wherein the solvent is selected from methanol, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone, ethyl acetate, or a combination thereof; (2) separating an extract and residue to obtain the residue; (3) extracting the residue of step (2) with water; (4) separating an water extract and residue to obtain the water extract; (5) concentrating, precipitating with ethanol; and separating to obtain the precipitate; and (6) optionally, dissolving and formulating the precipitate of step (5) into a solution.

5. The method according to claim 4, wherein the extractions of steps (1) and (3) are selected from soaking, decoction, refluxing, diacolation or a combination thereof.

6. The method according to claim 4, wherein the solvent used in step (6) is selected from ethanol, isopropanol, 1 -butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone, ethyl acetate or a combination thereof. 16

7. An extract of Eucommia ulmoides obtained according to the method of any one of claims 1 to 6.

8. The extract of Eucommia ulmoides according to claim 7, wherein the extract of Eucommia ulmoides is in a form selected from emulsion, solution, powder, ointment, film or cream.

9. A use of the extract of Eucommia ulmoides in the preparation of anti-aging, especially anti-photoaging cosmetics or medicaments.

10. The use according to claim 9, wherein the extract of Eucommia ulmoides is the extract of Eucommia ulmoides according to claim 7 or 8.

11. An anti-aging, especially anti-photoaging cosmetic or medicament, comprising the extract of Eucommia ulmoides.

12. The cosmetic or medicament according to claim 11, wherein the extract of Eucommia ulmoides is the extract of Eucommia ulmoides according to claim 7 or 8. 17