WO2011140856A1

WO2011140856A1 - Snow lotus extract, preparing method and application thereof

Info

Publication number: WO2011140856A1
Application number: PCT/CN2011/071680
Authority: WO
Inventors: 陈默; 熊卫国; 刘贝贝; 祝乐; 赵亚; 程康; 方程; 吕洛; 马来记
Original assignee: 上海家化联合股份有限公司
Priority date: 2010-05-13
Filing date: 2011-03-10
Publication date: 2011-11-17
Also published as: CN102240254A; AU2011252628A1

Abstract

A method for preparing snow lotus extract comprises a step of solvent extraction. The solvent is selected from water, methanol, ethanol, isopropyl alcohol, 1-butanol, ethylene glycol, 1,2-propylene glycol, 1,3-propylene glycol, 1,3-butylene glycol, acetone, ethyl acetate, or the combination thereof. The obtained snow lotus extract has the effects of anti-oxidation and resistance to UVA and UVB radiations. The snow lotus extract is used as an active ingredient in the manufacture of cosmetics or medicaments for skin anti-aging, especially anti-photoaging.

Description

Snow lotus extract, preparation method and application thereof

The invention relates to the field of natural medicinal chemistry and cosmetics, and in particular to a Chinese herbal medicine snow lotus for anti-aging, preferably anti-photoaging, in cosmetics or medicines. Background technique

Studies have shown that both free radicals and ultraviolet light are one of the factors of skin aging, and photoaging is mainly caused by the ultraviolet (UVB, 290~320nm) and long-wave ultraviolet (UVA, 320~400nm) in the dawn. Ultraviolet light has a significant effect on the aging of human skin. The risk of skin aging in high-exposure populations is twice as high as that in low-exposure populations, and the aging time is 10 years earlier. Long-term exposure to sunlight and ultraviolet light causes skin photoaging, which can lead to serious cosmetic problems. Therefore, it has received much attention in the industry in recent years. Anti-aging and anti-aging products have come out. Related research is like fire, but natural Chinese herbal medicine is resistant to light aging. The additive is still vacant. Summary of the invention

The invention aims to provide a method for preparing a snow lotus extract, the method comprising:

(1) solvent extraction, the solvent is selected from the group consisting of water, decyl alcohol, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, and 1,3-butanediol , acetone, ethyl acetate or a combination thereof.

Preferably, wherein the extraction of step (1) is selected from the group consisting of soaking, boiling, refluxing, percolating, or a combination thereof. Preferably, the preparation method further includes:

(2) separating the extract and the residue to obtain an extract;

(3) Optional solvent recovery;

(4) Optional dissolution or dilution.

Preferably, the solvent used in the step (4) is selected from the group consisting of water, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, and 1,3-butanediol. , acetone, ethyl acetate or a combination thereof.

Another object of the present invention is to provide a method for preparing another snow lotus extract, the method comprising:

(1) solvent extraction, the solvent is selected from the group consisting of decyl alcohol, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone , ethyl acetate or a combination thereof; (2) separating the extract and the residue to obtain a residue;

(3) The residue of the water extraction step (2).

Preferably, wherein the extraction of steps (1) and (3) is independently selected from the group consisting of soaking, boiling, refluxing, percolating, or a combination thereof.

Preferably, the preparation method further includes:

(4) separating the water extract and the residue to obtain a water extract;

(5) Concentration, ethanol alcohol precipitation, separation to obtain a precipitate;

(6) Alternatively, the precipitate of the step (5) is dissolved to prepare a solution.

Preferably, the solvent used in the step (6) is selected from the group consisting of ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, Acetone, ethyl acetate or a combination thereof.

Still another object of the present invention is to provide a snowdrop extract obtained according to the above production method of the present invention.

Preferably, the form of the snow lotus extract is selected from the group consisting of a solution, a powder, an ointment, a film or a cream.

The snowdrop extract obtained according to the method of the present invention can be directly used for preventing skin aging, preferably photoaging, and can also be used as an efficacy additive or an active ingredient for use in cosmetics or pharmaceuticals.

A further object of the present invention is to provide the use of a snow lotus extract in the preparation of a cosmetic or pharmaceutical product which is resistant to aging, preferably to photoaging.

Preferably, the snow lotus extract is the snow lotus extract obtained by the above method according to the present invention.

Still another object of the present invention is to provide an anti-aging, preferably photoaging-resistant cosmetic or cosmetic comprising a snow lotus extract.

In the present invention, snow lotus refers to the medicinal part of the flower of the plant snow lotus. Snow lotus is a herbaceous plant of the genus Chrysanthemum of the genus Chrysanthemum. It mainly includes Saussurea involucrata Kar.et Kir., Saussurea laniceps Hand.-Mazz., and S. gnaphaloides (Royle) ) Sch.-Bip Jellyfish Snowdrop Xwe^^z Ma; a'w.

Snow lotus is mainly produced in Sichuan, Yunnan, Tibet, Xinjiang, Ganzi, Qinghai, etc., from June to July, when the flowers are opened, the whole plant is removed, the soil is removed, dried, cut, and used.

Snow lotus heats and nourishes blood, balances yin and yang, and replenishes the benefits. More than a thousand years ago, "Tao Zang" called Snow Lotus For the "Millennium Fairy", "Reminiscence of the Old Poetry": "Snow Lotus, the millennium is not deep in the snow, it looks like a lotus flower, high can be made. Take the wine, multiply the spring. Cover the cathode and the meaning of the sun "The Compendium of the Compendium": "Snow Lotus, the north of the Ili and the Jinchuan where the ice is produced, the snow does not scatter in spring and summer, the grass in the snow, the lotus-like, the single stem, the lovely snow in the pavilion."

The snow lotus extract obtained by the method of the invention has the function of scavenging free radicals and anti-oxidation, and can help the skin to resist ultraviolet radiation of UVB and UVA, protect the skin, reduce cell damage, prevent reduction of dermal collagen, and effectively prevent skin aging. , especially photoaging, and no irritation to the skin. detailed description

Example 1

Take the snow lotus Saussurea laniceps Hand.-Mazz 100g, the force is 8 times the amount (multiple volume: the ratio of solvent volume to snow lotus weight, where the solvent volume is in ml, the weight of the snow lotus is g, the same below 75% ethanol (v/v) was immersed for 48 hours, filtered, and the filtrate was recovered to ethanol. The obtained extract was dissolved in water to prepare a solution containing a crude drug amount of 1000 mg/ml to obtain an extract A. Example 2

Take the S. gnaphaloides (Royle) Sch.-Bip. 100g, add 8 times the amount of sterol reflux for 2 hours, filter, and recover the sterol from the filtrate. The extract is dissolved in water to make a crude drug content of 500mg. /ml of the solution to obtain extract B. Example 3

Take the jellyfish S. medusa Maxim. 100g, extract with 10 times of water for about 2 hours, concentrate, filter, add water to the supernatant to make a solution containing 1000 mg/ml of crude drug to obtain extract (:. Example 4

Take the snow lotus Saussurea involucrata Kar. et Kir. 1 OOg, extract 6 times the amount of ethanol back to i3⁄4, filter, and then dilute with 10 times the amount of water for about 2 hours, concentrate, add 2 times the amount of ethanol precipitation, filter After that, the precipitate was dissolved in water to prepare a solution containing a crude drug amount of 500 mg/ml to obtain an extract 0. Example 5 DPPH experiment

Free radicals are a class of chemically active, extremely oxidizing chemicals. Ultraviolet light

UVA/UVB irradiation produces a large amount of free radicals, causing damage to the body and accelerating aging. Here, the in vitro antioxidant activity of the extracts in the examples was determined by DPPH radical scavenging. The extracts in the examples were each diluted to a concentration of 1% and 0.5% (v/v) with distilled water. Then, 2 ml of the extract of the example, 0.5 ml of a DPPH ethanol solution having a concentration of 500 μπιο1 ί, and 1.5 ml of water were added to the same test tube, shaken, and allowed to stand. After 30 minutes, the absorbance (OD) was measured at 517 nm, and correspondingly The mixture was used as a blank control. The clearance of DPPH from the extracts of the examples was calculated according to the following formula:

(— T - TO

Clearance rate (%) = I 1 - " I * 100 %

C~~ Where:

The absorbance of the extract of the T-DPPH+ example, TO - the absorbance of the extract + solvent, and the absorbance of the C-DPPH + solvent. The experimental results are shown in Table 1. Table 1 Evaluation of antioxidant capacity of snow lotus extract

The experimental results show that the Extract A of Example 1, the extract of Example 2, and the extract D of Example 4 have better anti-oxidation ability to scavenge free radicals in 1% and 0.5% concentrations in vitro, wherein 0.5% of free radicals The scavenging rate was above 10%, and the free radical scavenging rate of 1% concentration was above 30%. The extract D of Example 4 was the best. Example 6 Protection of fibroblast injury by UV irradiation

Skin photoaging is mainly caused by medium-wave ultraviolet (UVB, 290~320nm) and long-wave ultraviolet (UVA, 320~400nm) in sunlight, which has a significant effect on the aging of human skin. UVA/UVB irradiation causes damage to fibroblasts, causing damage to the skin and causing photoaging. Here, the UV-irradiation experiment was used to examine the photoaging effect of the herbal extract of the examples against UV-induced cell protection.

Take 0.5ml of the extract from the example, dilute with 4.5ml of PBS, and mix it with 1/10 concentration (v/v) for filtration. Then take 0.1ml and add 9,9ml of sterile PBS and finally dilute to 1/1000. Concentration (v/v). Skin fibroblasts (Fb) cultured in normal DMEM solution were used as a control group, and Fb cells irradiated with UVA (5-20 J/cm ² )/UVB UVB (20-80 J/cm ² ) were cultured in a normal DMEM solution. The Fb cells co-cultured with the 1/1000 concentration of the extract and the DMEM solution and irradiated with UVA/UVB were used as the dosing group. 24 hours after UV treatment, the absorbance (OD value) of each well was measured by CCK-8 using a microplate reader.

Each test was repeated three times.

The experimental results are shown in Tables 2 and 3.

Table 2 UVA irradiation

Table 3 UVB irradiation

Cell proliferation rate P value

Example 1 Extract A (93.27 ± 3.51)% <0.01

Example 2 Extract B (95.86 ± 2.85)% <0.01

Example 4 Extract D (97.54 ± 3.53)% <0.01

Control group (100±5.68)%

Model group (83.63±5.15)% <0.01 The experimental results show that the extract A of Example 1, the extract of Example 2, and the extract of Example 4

Both of them can reduce the damage of fibroblasts by UV irradiation, and the cell proliferation rate can be restored to more than 90%, and the effect of extract D in Example 4 is the best. Example 7 In vitro experiment of the effect of UV irradiation on dermal collagen

Take 0.5ml of the extract in the example, dilute with 4.5ml of PBS, and mix it with 1/10 concentration (v/v) for filtration. Then take 0.1ml and add 9.9ml of sterile PBS and finally dilute to 1/1000 concentration. (v/v)o Skin fibroblasts (Fb) cultured in normal DMEM solution were used as a control group, and cultured in normal DMEM solution and irradiated with UVA (5-20 J/cm ² ) UVB UVB (20-80 J/cm ² ). The Fb cells were a model group, and Fb cells co-cultured with 1/1000 concentration of the extract and DMEM solution and irradiated with UVA/UVB were used as the drug-added group. 24 h after the ultraviolet treatment, the cell supernatant was collected, and the hydroxyproline content in the culture supernatant was determined according to the method described in the hydroxyproline kit (Nanjing Jianjian).

The experimental results are shown in Tables 4 and 5.

Table 4 UVA irradiation

Collagen content Collagen change recovery ratio

(g/ml) rate (relative to model group) Example 1 Extract A 4.3 34.38%

Example 2 Extract B 4.9 53.13%

Example 4 Extract D 5.1 59.38%

Control group 8.2

Model group 3.2 Table 5 UVB irradiation

Collagen content Collagen change recovery ratio

(g/mi) rate (relative to the model group) Example 1 Extract A 4.8 26.32%

Example 2 Extract B 5.4 42.11%

Example 4 Extract D 6.6 73.68%

Control group 8, 2 Model group 3.8

The experimental results show that Example 1 Extract A, Example 2 Extract B, and Example 4 Extract D

Both can reduce the effect of UV irradiation on the collagen content of dermal fibroblasts, and the collagen can recover more than 25% compared with the model injury group, and the extract D of Example 4 works best, and can recover UVA 59.38%, UVB73 .68%.

Skin photoaging is mainly caused by medium-wave ultraviolet (UVB, 290-320nm) and long-wave ultraviolet (UVA, 320~400nm) in sunlight. UV rays can significantly promote the aging of human skin. UVA/UVB irradiation causes damage to fibroblasts, as well as reduction of dermal collagen, which can cause damage to the skin. The invention discloses that the cultured skin fibroblasts are irradiated with UVA/UVB of a certain intensity, and the snow lotus extract is incubated, which can effectively reduce the influence of UV irradiation on the proliferation of fibroblasts, and can effectively reduce the collagen change of dermal fibroblasts. Protect the skin, illustrating the snow lotus

Claims

Claim

A method for preparing a snow lotus extract, comprising:

The preparation method according to claim 1, wherein the extraction of the step (1) is selected from the group consisting of soaking, boiling, refluxing, percolating, or a combination thereof.

3. The preparation method according to claim 1 or 2, further comprising:

(2) separating the extract and the residue to obtain an extract;

(3) Optional solvent recovery;

(4) Optional dissolution or dilution.

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

5. A method for preparing a snow lotus extract, comprising:

(1) solvent extraction, the solvent is selected from the group consisting of decyl alcohol, ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, acetone , ethyl acetate or a combination thereof;

(2) separating the extract and the residue to obtain a residue;

(3) The residue of the water extraction step (2).

The preparation method according to claim 5, wherein the extraction of the steps (1) and (3) is independently selected from the group consisting of soaking, boiling, refluxing, percolating, or a combination thereof.

7. The preparation method according to claim 5 or 6, further comprising:

(4) separating the water extract and the residue to obtain a water extract;

The method according to claim 7, wherein the solvent used in the step (6) is selected from the group consisting of ethanol, isopropanol, 1-butanol, ethylene glycol, 1,2-propanediol, and 1,3-propanediol. , 1,3-butanediol, acetone, ethyl acetate or a combination thereof.

9. A snow lotus extract obtained by the production method according to any one of claims 1-8.

The snow lotus extract according to claim 9, wherein the shape of the snow lotus extract The formula is selected from the group consisting of a solution, a powder, an ointment, a film or a cream.

11. The use of snow lotus extract in the preparation of anti-aging, preferably anti-photoaging cosmetics or pharmaceuticals.

The use according to claim 11, wherein the snow lotus extract is the snow lotus extract according to claim 9 or 10.

13. An anti-aging, preferably photoaging-resistant cosmetic or pharmaceutical product comprising a snow lotus extract.

The cosmetic according to claim 13, wherein the snow lotus extract is the snow lotus extract according to claim 9 or 10.