CN118436556A - A resveratrol supramolecule and its preparation method and application in the preparation of cosmetics - Google Patents

Abstract

The present invention relates to the field of biomedical technology, and specifically discloses a method for preparing a resveratrol supramolecule, which comprises the following steps: S11. dissolving a cyclodextrin derivative in a solvent, then adding resveratrol, and stirring at 40-80°C for 1-3h to obtain a mixed solution; S12. spray drying the mixed solution to obtain the resveratrol supramolecule. Studies have shown that the resveratrol supramolecule prepared by the method of the present invention has a better antioxidant effect than an equal amount of resveratrol; in addition, compared with resveratrol, it can also reduce skin irritation. Further studies have shown that the composition obtained by mixing the resveratrol supramolecule and the root extract of Polygonum cuspidatum also has significant or even excellent anti-Propionibacterium acnes. Therefore, the resveratrol supramolecule and/or the composition of the present invention as an active ingredient for preparing cosmetics or drugs has important application value.

Description

A resveratrol supramolecule and its preparation method and application in the preparation of cosmetics

Technical Field

The invention relates to the field of biotechnology, and in particular to a resveratrol supramolecule and a preparation method thereof and application thereof in the preparation of cosmetics.

Background technique

Everyone loves beauty, and healthy and youthful skin is what everyone pursues. Anti-aging products such as anti-wrinkle and firming products occupy an important position in the cosmetics market, and rank first in market share among various cosmetic products. Ultraviolet exposure causes excessive reactive oxygen free radicals (ROS) in the skin, which can lead to the loss of skin collagen, a decrease in fibroblasts, loss of skin elasticity, and increased wrinkles, which is the so-called skin photoaging. Adding appropriate antioxidants to cosmetics can help the body remove excess ROS. Finding antioxidants that are efficient, safe, and very convenient to use in cosmetic formulas is an important means to combat photoaging.

Resveratrol is a non-flavonoid polyphenol compound, with the chemical name of 3,4',5-trihydroxy-1,2-diphenylethylene, mainly derived from plants such as grapes, knotweed, peanuts, Veratrum, and mulberries. In 2003, Harvard University professor David Sinclair and his team found that resveratrol can activate acetylase and increase the life span of yeast; Howitz et al. found that resveratrol can be used as the strongest activator of silent information regulation 2homolog1 (SIRT1), which can simulate the anti-aging response of calorie restriction (CR) and participate in the regulation of the average life span of organic organisms. CR is a strong inducer of SIRT1, which can increase the expression of SIRT1 in organs and tissues such as brain, heart, intestine, kidney, muscle and fat. CR can cause physiological changes that delay aging and prolong life, the most significant of which can prolong life by 50%. Studies have also found that resveratrol has a significant inhibitory effect on various tumor cells such as rat hepatocellular carcinoma, breast cancer, colon cancer, gastric cancer, leukemia, etc. It also has some other biological activities such as antibacterial, antioxidant, immunomodulatory, and anti-asthmatic. Resveratrol has been widely used in the fields of food and health products. In recent years, it has also been used as an antioxidant in the field of cosmetics. Although resveratrol has strong antioxidant activity, its wide application in cosmetics is limited by its defects such as poor solubility in water and certain irritation. Therefore, using resveratrol as a raw material to develop more active ingredients for drugs or cosmetics has important application value.

The concept of supramolecules was proposed by Ryan, the winner of the 1987 Nobel Prize in Chemistry. Supramolecules refer to complex molecules composed of two or more molecules bound together in a non-covalent bond. They have been widely used in the fields of medicine, food, environment, materials, etc., and have also been introduced into the field of cosmetics in recent years. The annular inner cavity of cyclodextrin and its derivatives composed of multiple glucose units can self-assemble with guest molecules through host-guest interactions to obtain supramolecules, which is a class of subjects widely used in the field of supramolecules. The inner cavity of cyclodextrin and its derivatives is between 0.47-0.75nm, which can be used as a molecular capsule. The inner cavity of the molecular capsule is hydrophobic and the outer shell is hydrophilic, which can well load molecules with poor solubility in water, protect the guest molecules from oxidation and discoloration, and effectively increase their water solubility. It can also play a role in sustained release, promote skin penetration, increase bioavailability, and reduce the irritation of guest molecules. However, there is no research report on the formation of supramolecules by resveratrol and cyclodextrin derivatives in the prior art.

Summary of the invention

In order to overcome at least one of the technical problems existing in the prior art, the present invention provides a resveratrol supramolecule and a preparation method thereof and an application thereof in the preparation of cosmetics.

The present invention first provides a method for preparing resveratrol supramolecules, which comprises the following steps:

S11. dissolving a cyclodextrin derivative in a solvent, then adding resveratrol, and stirring at 40 to 80° C. for 1 to 3 hours to obtain a mixed solution;

S12. The mixed solution is spray-dried to obtain the resveratrol supramolecule.

The present invention provides a resveratrol supramolecule prepared by a completely new method; studies have shown that the resveratrol supramolecule prepared by the method of the present invention has a better antioxidant effect than an equal amount of resveratrol; in addition, compared with resveratrol, it can also reduce skin irritation.

Preferably, in step S11, the weight ratio of the cyclodextrin derivative to resveratrol and the solvent is 70-99:1-30:200-400.

Most preferably, in step S11, the weight ratio of the cyclodextrin derivative to resveratrol and the solvent is 90:10:270.

Preferably, the cyclodextrin derivative in step S11 is selected from one or more of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxypropyl α-cyclodextrin, hydroxypropyl β-cyclodextrin and hydroxypropyl γ-cyclodextrin.

The present invention also provides a resveratrol supramolecule prepared by the above preparation method.

The present invention also provides a composition comprising the above-mentioned resveratrol supramolecule.

Preferably, the composition comprises the above-mentioned resveratrol supramolecule and Polygonum cuspidatum root extract.

The inventors further found in the study that the composition obtained by mixing resveratrol supramolecules and Polygonum cuspidatum root extract has an antibacterial effect on Propionibacterium acnes. Since Propionibacterium acnes is the main bacteria that causes acne and pimples, the composition composed of resveratrol supramolecules and Polygonum cuspidatum root extract can also play a role in treating acne or removing pimples.

Preferably, the weight ratio of resveratrol supramolecule to Polygonum cuspidatum root extract is 1 to 3:1;

Most preferably, the weight ratio of resveratrol supramolecule to Polygonum cuspidatum root extract is 2:1.

Preferably, the Polygonum cuspidatum root extract is prepared by the following method:

Take the root of Polygonum cuspidatum, extract it with a solvent to obtain an extract; concentrate and dry the extract to obtain a Polygonum cuspidatum root solvent extract; take the Polygonum cuspidatum root solvent extract to obtain the Polygonum cuspidatum root extract.

Preferably, the solvent is a mixed solvent consisting of acetone and methanol.

More preferably, the volume ratio of acetone to methanol is 1:2-4.

Most preferably, the volume ratio of acetone to methanol is 1:3.

Preferably, the usage ratio of Polygonum cuspidatum root to solvent is 1kg:8-20L.

Most preferably, the usage ratio of Polygonum cuspidatum root and solvent is 1kg:15L.

Preferably, the method for preparing the Polygonum cuspidatum root extract further comprises the following steps:

The solvent extract of Polygonum cuspidatum root is placed on a macroporous resin column, and impurities are first eluted with 4 to 8 times the column volume of an ethanol aqueous solution with a volume fraction of 36 to 39%; then it is eluted with 4 to 8 times the column volume of an ethanol aqueous solution with a volume fraction of 53 to 55%, and the eluate eluted with the 53 to 55% ethanol aqueous solution is collected, and after concentrating and drying, the macroporous resin elution portion of Polygonum cuspidatum root is obtained; the Polygonum cuspidatum root extract is obtained by taking the macroporous resin elution portion of Polygonum cuspidatum root.

Most preferably, the method for preparing the Polygonum cuspidatum root extract further comprises the following steps:

The solvent extract of Polygonum cuspidatum root is placed on a macroporous resin column, and impurities are first eluted with 6 times the column volume of 38% ethanol aqueous solution; then it is eluted with 6 times the column volume of 54% ethanol aqueous solution, and the eluate eluted with the 54% ethanol aqueous solution is collected, concentrated and dried to obtain the Polygonum cuspidatum root macroporous resin elution part; the Polygonum cuspidatum root macroporous resin elution part is taken to obtain the Polygonum cuspidatum root extract.

Preferably, the macroporous resin column is a D101 macroporous resin column.

The inventors found in their research that the composition obtained by mixing resveratrol supramolecules with the macroporous resin elution part of Polygonum cuspidatum root prepared by the above-mentioned macroporous resin elution conditions has a significantly higher anti-Propionibacterium acnes effect than the composition obtained by mixing resveratrol supramolecules with the solvent extract of Polygonum cuspidatum root without macroporous resin elution.

In addition, the inventors need to emphasize that the elution conditions of the macroporous resin column of the present invention are very critical. Only when the resveratrol supramolecule is mixed with the macroporous resin elution part of the Polygonum cuspidatum root prepared by the macroporous resin column elution conditions of the present invention, the anti-Propionibacterium acnes effect of the composition can be further significantly higher than the composition obtained by mixing the resveratrol supramolecule with the Polygonum cuspidatum root solvent extract without macroporous resin elution; however, the anti-Propionibacterium acnes effect of the composition obtained by mixing the resveratrol supramolecule with the macroporous resin elution part of the Polygonum cuspidatum root prepared by other macroporous resin column elution conditions cannot be further significantly higher than the composition obtained by mixing the resveratrol supramolecule with the Polygonum cuspidatum root solvent extract without macroporous resin elution.

Preferably, the method for preparing the Polygonum cuspidatum root extract further comprises the following steps:

The elution part of the Polygonum cuspidatum root macroporous resin is placed on a silica gel column, and impurities are first eluted with a mixed solvent consisting of chloroform and methanol in a volume ratio of 95-98:5-2 for 3-5 times the column volume; then, it is eluted with a mixed solvent consisting of chloroform and methanol in a volume ratio of 83-85:17-15 for 4-6 times the column volume, and the eluate eluted with the mixed solvent consisting of chloroform and methanol in a volume ratio of 83-85:17-15 is collected, and the elution part of the Polygonum cuspidatum root silica gel column is obtained after concentration and drying; the Polygonum cuspidatum root silica gel column elution part is taken to obtain the Polygonum cuspidatum root extract.

The inventors found in their research that the composition obtained by mixing resveratrol supramolecules with the polygonum cuspidatum root silica gel column elution fraction prepared by the above-mentioned silica gel column elution conditions has a further significantly higher anti-Propionibacterium acnes effect than the composition obtained by mixing resveratrol supramolecules with the polygonum cuspidatum root macroporous resin elution fraction.

Similarly, the inventors need to emphasize that the silica gel column elution conditions of the present invention are very critical. Only when the resveratrol supramolecule is mixed with the polygonum cuspidatum root silica gel column elution fraction prepared by the silica gel column elution conditions of the present invention, the anti-Propionibacterium acnes effect of the composition can be further significantly higher than the composition obtained by mixing the resveratrol supramolecule with the polygonum cuspidatum root macroporous resin elution fraction; however, the anti-Propionibacterium acnes effect of the composition obtained by mixing the resveratrol supramolecule with the polygonum cuspidatum root silica gel column elution fraction prepared by other silica gel column elution conditions cannot be further significantly higher than the composition obtained by mixing the resveratrol supramolecule with the polygonum cuspidatum root macroporous resin elution fraction.

Preferably, the composition is a composition having an anti-Propionibacterium acnes effect.

The present invention also provides a use of the above-mentioned resveratrol supramolecule or composition in preparing a product with whitening, anti-oxidation and/or acne-removing effects.

Preferably, the composition is used in the preparation of a product having an anti-Propionibacterium acnes effect.

Preferably, the product is a cosmetic or a medicine.

The present invention also provides an application of the resveratrol supramolecule or composition in the preparation of cosmetics.

Preferably, the cosmetics include but are not limited to essences, creams, facial masks, lotions, gels, and sprays.

Beneficial effects: The present invention provides a resveratrol supramolecule prepared by a completely new method; studies have shown that the resveratrol supramolecule prepared by the method of the present invention has a better antioxidant effect than the same amount of resveratrol; in addition, compared with resveratrol, it can also reduce skin irritation. Further studies have shown that the composition obtained by mixing the resveratrol supramolecule and the root extract of Polygonum cuspidatum has significant or even excellent anti-Propionibacterium acnes. Therefore, the resveratrol supramolecule and/or composition of the present invention as an active ingredient for the preparation of cosmetics or drugs has important application value.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 is the H NMR spectrum of pure resveratrol.

FIG. 2 is a hydrogen nuclear magnetic resonance spectrum of the resveratrol supramolecule of the present invention.

Figure 3 is the H NMR spectrum of pure hydroxypropyl-β-cyclodextrin.

Detailed ways

The present invention is further explained below in conjunction with specific examples, but the examples do not limit the present invention in any form.

Example 1 Preparation of resveratrol supramolecular

90 g of hydroxypropyl-β-cyclodextrin was added to 270 g of water and heated to 70° C. After it was completely dissolved, 10 g of resveratrol was added, and then stirred at this temperature for 2 hours. The mixture was spray-dried under nitrogen protection, the inlet air temperature was controlled at 125° C., and the injection speed was maintained at 15 ml/min. The spray-dried product was collected to obtain the resveratrol supramolecule.

Resveratrol: ¹ HNMR (400 MHz, CD ₃ OD): δ7.35 (d, J＝8.4 Hz, 2H), 6.96 (d, J＝16.4 Hz, 1H), 6.82-6.75 (m, 3H), 6.45-6.44 (m, 2H), 6.16-6.15 (m, 1H). See Figure 1 for the spectrum.

Resveratrol supramolecular: ¹ HNMR (400MHz, CD ₃ OD): δ7.37 (d, J＝8.4Hz, 2H), 6.96 (d, J＝16.4Hz, 1H), 6.82-6.76 (m, 3H). The hydrogen chemical shift of hydroxypropyl-β-cyclodextrin is not listed, and the specific spectra are shown in Figures 2 and 3.

Comparing the H NMR spectra of resveratrol monomer and supramolecule, changes occurred in the low-field region, indicating that resveratrol entered the inner cavity of hydroxypropyl-β-cyclodextrin, which can confirm the formation of supramolecule.

Experimental Example 1 Human Patch Test

Resveratrol and the resveratrol supramolecule prepared according to the method of Example 1 were respectively prepared into creams A and B, both of which contained the same concentration of resveratrol and the same other ingredients. The formula compositions are shown in Table 1. Table 4: Compositions of creams A and B.

Table 1. Composition of cream by weight

30 volunteers aged 22-49 were selected, including 9 males and 21 females, with an average age of 28±6.1 years old, meeting the selection criteria for subject volunteers. About 0.020-0.025 ml of cream A and B were added to the spot tester, and then the spot tester with the sample was applied to the back of the subject with a non-irritating tape, and gently pressed with the palm of the hand to evenly apply it to the skin for 24 hours; the test spot tester was removed, and the test residue on the test site was gently wiped off with a moistened cotton ball. After 0.5 hours, the skin reaction was observed after the indentation disappeared. If the result is negative, it was observed again at 24 hours and 48 hours after the patch was removed. The reaction results were recorded according to the grading standard, as shown in Table 2.

Table 2. Human patch test

The results in Table 1 show that the cream prepared with resveratrol monomer has certain irritation to human skin, while the cream prepared with resveratrol supramolecule prepared according to the method of Example 1 with the same resveratrol content has no irritation to human skin, indicating that the molecular capsule formed by the inner cavity of the main molecule hydroxypropyl β-cyclodextrin in the resveratrol supramolecule of the present invention can well protect the guest molecule resveratrol, reduce the direct contact between resveratrol and human skin, and thus reduce the irritation of resveratrol to the human body.

Experimental Example 2 Zebrafish Activity Test

The genetic similarity between zebrafish and humans is as high as 87%. In 2003, the U.S. Institute of Health listed zebrafish as the third largest model organism after rats and mice, and it is known as the "little white mouse" in the water. Its physiology, developmental metabolism are highly similar to those of mammals, and its skin structure is highly similar to that of humans. Zebrafish skin also has a basal layer, spinous layer, granular layer, clear layer and epidermal keratinocyte layer, as well as the same structure of human skin as the lamina propria, basal granules, melanocytes, blood vessels and subcutaneous fat cells, etc. Therefore, zebrafish can be used in acute toxic substance testing and efficacy testing, and has been widely recognized in the cosmetics industry.

The ROS regulation mechanism in zebrafish is the same as that in humans. ROS in zebrafish embryos can be monitored by live staining with the fluorescent probe 2',7'-dichlorofluorescein diacetate (H2DCFDA), which is marked as green fluorescence. Zebrafish embryos are small, which makes it convenient to take pictures under a microscope. Software analysis is used to quantitatively read the green fluorescence signal in zebrafish embryos that increases with the increase of ROS. The changes in ROS levels in fish embryos in the treatment group and the blank control group are tested and compared, and the ability of hydroxytyrosol to remove ROS is calculated.

The method of the detection experiment is to expose 24 48-hour-old zebrafish embryos to the corresponding resveratrol or resveratrol supramolecular solution, and set up a blank control group. After 24 hours of exposure, the fish embryos were stained with H2DCFDA, and the ROS signal intensity was measured by fluorescence photography and statistical analysis. The final reactive oxygen species (ROS) clearance rate was calculated according to the following formula:

Clearance = [(C-T)/C] X 100%

In the formula, T is the average value of the “average signal intensity” of ROS in the fish embryos in the test substance treatment group; C is the average value of the “average signal intensity” of ROS in the fish embryos in the blank control group. The calculation results are shown in Table 3.

Table 3. Zebrafish activity test

From the experimental results in Table 3, it can be seen that the ROS scavenging rate of resveratrol monomer on zebrafish embryos can reach 90% when the test concentration is 1g/L, and the effect is significant; but when the test concentrations are 0.1g/L and 0.005g/L, the ROS scavenging rate drops to 63% and 19%; while the ROS scavenging rate of resveratrol hydroxypropyl β-cyclodextrin supramolecular on zebrafish embryos can reach 99% when the test concentration is 0.5g/L (the supramolecular contains 10% resveratrol, and the actual test concentration of resveratrol is 0.05g/L); the ROS scavenging rate of resveratrol supramolecular on zebrafish embryos can still reach 60% when the test concentration of resveratrol supramolecular is 0.25g/L (the actual test concentration of resveratrol is 0.025g/L). The results clearly show that at the same concentration of resveratrol, the resveratrol supramolecular of the present invention has better biological activity than resveratrol monomer, indicating that the molecular capsule formed by the supramolecular host molecule hydroxypropyl β-cyclodextrin can not only well protect the guest molecule resveratrol, but also play a role in delivery and sustained release, which can greatly improve the bioavailability of the guest molecule, so that the guest molecule at the same concentration can exert better biological activity in the supramolecular state and show a better ROS clearance rate.

Example 2 Preparation of the composition

The resveratrol supramolecule prepared according to the method of Example 1 and the Polygonum cuspidatum root extract are uniformly mixed in a weight ratio of 2:1 to obtain the composition;

The polygonum cuspidatum root extract is prepared by the following method: taking dried polygonum cuspidatum root, performing reflux extraction with a solvent at 56° C. for 1.5 hours to obtain an extract; concentrating and drying the extract to obtain a polygonum cuspidatum root solvent extract; wherein the amount ratio of dried polygonum cuspidatum root to solvent is 1kg:15L; the solvent is composed of acetone and methanol in a volume ratio of 1:3;

The Polygonum cuspidatum root extract is obtained by taking the Polygonum cuspidatum root solvent extract.

Example 3 Preparation of the composition

The resveratrol supramolecule prepared according to the method of Example 1 and the Polygonum cuspidatum root extract are uniformly mixed in a weight ratio of 2:1 to obtain the composition;

The Polygonum cuspidatum root extract is prepared by the following method:

S21. Take dried Polygonum cuspidatum root, reflux extract with solvent at 56°C for 1.5h to obtain an extract; concentrate and dry the extract to obtain a Polygonum cuspidatum root solvent extract; wherein the amount ratio of dried Polygonum cuspidatum root to solvent is 1kg:15L; the solvent is composed of acetone and methanol in a volume ratio of 1:3;

S22. The root of Polygonum cuspidatum solvent extract was loaded onto a macroporous resin column (the macroporous resin column was filled with 50 times the weight of the root of Polygonum cuspidatum solvent extract of D101 macroporous resin), and impurities were first eluted with 6 times the volume of the column with a volume fraction of 38% ethanol aqueous solution; then eluted with 6 times the volume of the column with a volume fraction of 54% ethanol aqueous solution, and the eluate eluted with a volume fraction of 54% ethanol aqueous solution was collected, and the eluate was concentrated and dried to obtain the root of Polygonum cuspidatum macroporous resin elution part;

The Polygonum cuspidatum root extract is obtained by taking the macroporous resin elution part of the Polygonum cuspidatum root.

Example 4 Preparation of the composition

The resveratrol supramolecule prepared according to the method of Example 1 and the Polygonum cuspidatum root extract are uniformly mixed in a weight ratio of 2:1 to obtain the composition;

The Polygonum cuspidatum root extract is prepared by the following method:

S21. Take dried Polygonum cuspidatum root, reflux extract with solvent at 56°C for 1.5h to obtain an extract; concentrate and dry the extract to obtain a Polygonum cuspidatum root solvent extract; wherein the amount ratio of dried Polygonum cuspidatum root to solvent is 1kg:15L; the solvent is composed of acetone and methanol in a volume ratio of 1:3;

S22. The root of Polygonum cuspidatum solvent extract was loaded onto a macroporous resin column (the macroporous resin column was filled with 50 times the weight of the root of Polygonum cuspidatum solvent extract of D101 macroporous resin), and impurities were first eluted with 6 times the volume of the column with a volume fraction of 38% ethanol aqueous solution; then eluted with 6 times the volume of the column with a volume fraction of 54% ethanol aqueous solution, and the eluate eluted with a volume fraction of 54% ethanol aqueous solution was collected, and the eluate was concentrated and dried to obtain the root of Polygonum cuspidatum macroporous resin elution part;

S23. The elution site of Polygonum cuspidatum macroporous resin was placed on a silica gel column (the silica gel column was filled with 200-300 mesh silica gel 30 times the weight of the elution site of Polygonum cuspidatum macroporous resin), and impurities were removed by elution with a mixed solvent of chloroform and methanol in a volume ratio of 96:4 for 4 times the column volume; then eluted with a mixed solvent of chloroform and methanol in a volume ratio of 84:16 for 5 times the column volume, and the eluate was collected by volume ratio of 84:16 for chloroform and methanol. The eluate was concentrated and dried to obtain the elution site of Polygonum cuspidatum silica gel column;

The Polygonum cuspidatum root extract is obtained by taking the elution portion of the Polygonum cuspidatum root silica gel column.

Comparative Example 1 Preparation of the composition

The resveratrol supramolecule prepared according to the method of Example 1 and the Polygonum cuspidatum root extract are uniformly mixed in a weight ratio of 2:1 to obtain the composition;

The Polygonum cuspidatum root extract is prepared by the following method:

S21. Take dried Polygonum cuspidatum root, reflux extract with solvent at 56°C for 1.5h to obtain an extract; concentrate and dry the extract to obtain a Polygonum cuspidatum root solvent extract; wherein the amount ratio of dried Polygonum cuspidatum root to solvent is 1kg:15L; the solvent is composed of acetone and methanol in a volume ratio of 1:3;

S22. The root of Polygonum cuspidatum solvent extract was loaded onto a macroporous resin column (the macroporous resin column was filled with 50 times the weight of the root of Polygonum cuspidatum solvent extract of D101 macroporous resin), and impurities were first eluted with 6 times the volume of the column with a volume fraction of 10% ethanol aqueous solution; then eluted with 6 times the volume of the column with a volume fraction of 30% ethanol aqueous solution, and the eluate eluted with a volume fraction of 30% ethanol aqueous solution was collected, and the eluate was concentrated and dried to obtain the root of Polygonum cuspidatum macroporous resin elution part;

The Polygonum cuspidatum root extract is obtained by taking the macroporous resin elution part of the Polygonum cuspidatum root.

The difference between Comparative Example 1 and Example 3 is that the elution conditions of the macroporous resin column are different.

Comparative Example 2 Preparation of the composition

The resveratrol supramolecule prepared according to the method of Example 1 and the Polygonum cuspidatum root extract are uniformly mixed in a weight ratio of 2:1 to obtain the composition;

The Polygonum cuspidatum root extract is prepared by the following method:

S21. Take dried Polygonum cuspidatum root, reflux extract with solvent at 56°C for 1.5h to obtain an extract; concentrate and dry the extract to obtain a Polygonum cuspidatum root solvent extract; wherein the amount ratio of dried Polygonum cuspidatum root to solvent is 1kg:15L; the solvent is composed of acetone and methanol in a volume ratio of 1:3;

S22. The root of Polygonum cuspidatum solvent extract was loaded onto a macroporous resin column (the macroporous resin column was filled with 50 times the weight of the root of Polygonum cuspidatum solvent extract of D101 macroporous resin), and impurities were first eluted with 6 times the volume of the column with a volume fraction of 55% ethanol aqueous solution; then eluted with 6 times the volume of the column with a volume fraction of 75% ethanol aqueous solution, and the eluate eluted with a volume fraction of 75% ethanol aqueous solution was collected, and the eluate was concentrated and dried to obtain the root of Polygonum cuspidatum macroporous resin elution part;

The Polygonum cuspidatum root extract is obtained by taking the macroporous resin elution part of the Polygonum cuspidatum root.

The difference between Comparative Example 2 and Example 3 is that the elution conditions of the macroporous resin column are different.

Comparative Example 3 Preparation of Composition

The resveratrol supramolecule prepared according to the method of Example 1 and the Polygonum cuspidatum root extract are uniformly mixed in a weight ratio of 2:1 to obtain the composition;

The Polygonum cuspidatum root extract is prepared by the following method:

S21. Take dried Polygonum cuspidatum root, reflux extract with solvent at 56°C for 1.5h to obtain an extract; concentrate and dry the extract to obtain a Polygonum cuspidatum root solvent extract; wherein the amount ratio of dried Polygonum cuspidatum root to solvent is 1kg:15L; the solvent is composed of acetone and methanol in a volume ratio of 1:3;

S22. The root of Polygonum cuspidatum solvent extract was loaded onto a macroporous resin column (the macroporous resin column was filled with 50 times the weight of the root of Polygonum cuspidatum solvent extract of D101 macroporous resin), and impurities were first eluted with 6 times the volume of the column with a volume fraction of 38% ethanol aqueous solution; then eluted with 6 times the volume of the column with a volume fraction of 54% ethanol aqueous solution, and the eluate eluted with a volume fraction of 54% ethanol aqueous solution was collected, and the eluate was concentrated and dried to obtain the root of Polygonum cuspidatum macroporous resin elution part;

S23. The elution site of Polygonum cuspidatum macroporous resin was placed on a silica gel column (the silica gel column was filled with 200-300 mesh silica gel 30 times the weight of the elution site of Polygonum cuspidatum macroporous resin), and impurities were removed by elution with a mixed solvent of chloroform and methanol in a volume ratio of 99:1 for 4 times the column volume; then eluted with a mixed solvent of chloroform and methanol in a volume ratio of 90:10 for 5 times the column volume, and the eluate eluted by a mixed solvent of chloroform and methanol in a volume ratio of 90:10 was collected, and the eluate was concentrated and dried to obtain the elution site of Polygonum cuspidatum silica gel column;

The Polygonum cuspidatum root extract is obtained by taking the elution portion of the Polygonum cuspidatum root silica gel column.

The difference between Comparative Example 3 and Example 4 is that the elution conditions of the silica gel column are different.

Preparation of the composition of Comparative Example 4

The resveratrol supramolecule prepared according to the method of Example 1 and the Polygonum cuspidatum root extract are uniformly mixed in a weight ratio of 2:1 to obtain the composition;

The Polygonum cuspidatum root extract is prepared by the following method:

S21. Take dried Polygonum cuspidatum root, reflux extract with solvent at 56°C for 1.5h to obtain an extract; concentrate and dry the extract to obtain a Polygonum cuspidatum root solvent extract; wherein the amount ratio of dried Polygonum cuspidatum root to solvent is 1kg:15L; the solvent is composed of acetone and methanol in a volume ratio of 1:3;

S22. The root of Polygonum cuspidatum solvent extract was loaded onto a macroporous resin column (the macroporous resin column was filled with 50 times the weight of the root of Polygonum cuspidatum solvent extract of D101 macroporous resin), and impurities were first eluted with 6 times the volume of the column with a volume fraction of 38% ethanol aqueous solution; then eluted with 6 times the volume of the column with a volume fraction of 54% ethanol aqueous solution, and the eluate eluted with a volume fraction of 54% ethanol aqueous solution was collected, and the eluate was concentrated and dried to obtain the root of Polygonum cuspidatum macroporous resin elution part;

S23. The elution site of Polygonum cuspidatum macroporous resin was placed on a silica gel column (the silica gel column was filled with 200-300 mesh silica gel 30 times the weight of the elution site of Polygonum cuspidatum macroporous resin), and impurities were removed by elution with a mixed solvent of chloroform and methanol in a volume ratio of 85:15 for 4 times the column volume; then eluted with a mixed solvent of chloroform and methanol in a volume ratio of 70:30 for 5 times the column volume, and the eluate was collected by volume in a mixed solvent of chloroform and methanol in a volume ratio of 70:30, and concentrated and dried to obtain the elution site of Polygonum cuspidatum silica gel column;

The Polygonum cuspidatum root extract is obtained by taking the elution portion of the Polygonum cuspidatum root silica gel column.

The difference between Comparative Example 4 and Example 4 is that the elution conditions of the silica gel column are different.

Experimental Example 3

The samples to be tested were diluted into sample solutions with concentrations of 256, 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, and 0.125 μg/ml according to the doubling dilution method; the samples to be tested were the compositions prepared in Examples 2 to 4 and Comparative Examples 1 to 4, respectively.

A 1×10 ⁷ cfu/ml Propionibacterium acnes bacterial solution was added to a 96-well plate, and the test sample solutions of the above concentrations were added to the experimental groups in sequence; then the 96-well plate was placed in a cell culture incubator and cultured at 37°C for 24 hours; in addition, a blank control group without the test sample solution was set up; 3 replicate wells were set up for each group.

After the culture was completed, the absorbance value A at 600 nm in each well was detected by an ELISA instrument, and the inhibition rate of the cells under each test sample concentration condition in each group was calculated according to the absorbance value A; inhibition rate = (1-A _{test group} /A _{blank control group} ) × 100%, and the IC ₅₀ value was further calculated using SPSS statistical software; the results are shown in Table 4.

Table 4. Experimental results of the composition of the present invention against Propionibacterium acnes

From the experimental results in Table 4, it can be seen that the IC ₅₀ value of the composition prepared in Example 2 against Propionibacterium acnes is 25.3 μg/ml; this shows that the composition obtained by mixing resveratrol supramolecules and Polygonum cuspidatum root extract (Polygonum cuspidatum root solvent extract) has an antibacterial effect on Propionibacterium acnes.

It can be seen from the experimental results in Table 4 that the IC ₅₀ value of the composition prepared in Example 3 against Propionibacterium acnes is significantly lower than that in Example 2; this indicates that the composition obtained by mixing the resveratrol supramolecule with the Polygonum cuspidatum root macroporous resin elution fraction prepared by the macroporous resin elution conditions of the present invention has a significantly higher anti-Propionibacterium acnes effect than the composition obtained by mixing the resveratrol supramolecule with the Polygonum cuspidatum root solvent extract not eluted with the macroporous resin.

It can be seen from the experimental results in Table 4 that the IC ₅₀ value of the composition prepared in Comparative Example 1 against Propionibacterium acnes is lower than that in Example 2, but the reduction is not significant; the IC ₅₀ value of the composition prepared in Comparative Example 2 against Propionibacterium acnes is significantly increased compared with that in Example 2. This shows that: the elution conditions of the macroporous resin column of the present invention are very critical, and the types and contents of the effective ingredients in the macroporous resin elution parts of the Polygonum cuspidatum root obtained by eluting with different macroporous resin elution conditions are completely different; this leads to completely different effects against Propionibacterium acnes; the above experimental results show that only when the resveratrol supramolecule is mixed with the macroporous resin elution part of the Polygonum cuspidatum root prepared by the macroporous resin column elution conditions of the present invention, the anti-Propionibacterium acnes effect of the composition can be further significantly higher than that of the composition obtained by mixing the resveratrol supramolecule with the Polygonum cuspidatum root solvent extract without macroporous resin elution; however, the anti-Propionibacterium acnes effect of the composition obtained by mixing the resveratrol supramolecule with the macroporous resin elution part of the Polygonum cuspidatum root prepared by other macroporous resin column elution conditions cannot be further significantly higher than that of the composition obtained by mixing the resveratrol supramolecule with the Polygonum cuspidatum root solvent extract without macroporous resin elution.

It can be seen from the experimental results in Table 4 that the IC ₅₀ value of the composition prepared in Example 4 against Propionibacterium acnes is further significantly lower than that in Example 3; this indicates that the composition obtained by mixing the resveratrol supramolecule with the polygonum cuspidatum root silica gel column elution fraction prepared by the silica gel column elution conditions of the present invention has an anti-Propionibacterium acnes effect that is further significantly higher than the composition obtained by mixing the resveratrol supramolecule with the polygonum cuspidatum root macroporous resin elution fraction.

It can be seen from the experimental results in Table 1 that the IC ₅₀ value of the composition prepared in Comparative Example 3 against Propionibacterium acnes is lower than that in Example 3, but the reduction is not significant; the IC ₅₀ value of the composition prepared in Comparative Example 4 against Propionibacterium acnes is not lowered but increased compared with that in Example 3. This shows that: the silica gel column elution conditions of the present invention are very critical, and the types and contents of the effective ingredients in the polygonum cuspidatum root silica gel column elution parts obtained by eluting with different silica gel column elution conditions are completely different; this leads to completely different effects against Propionibacterium acnes; the above experimental results show that only when the resveratrol supramolecule is mixed with the polygonum cuspidatum root silica gel column elution part prepared by the silica gel column elution conditions of the present invention, the anti-Propionibacterium acnes effect of the composition can be further significantly higher than that of the composition obtained by mixing the resveratrol supramolecule with the polygonum cuspidatum root macroporous resin elution part; however, the anti-Propionibacterium acnes effect of the composition obtained by mixing the resveratrol supramolecule with the polygonum cuspidatum root silica gel column elution part prepared by other silica gel column elution conditions cannot be further significantly higher than that of the composition obtained by mixing the resveratrol supramolecule with the polygonum cuspidatum root macroporous resin elution part.

Claims (10)

1. A method for preparing resveratrol supramolecules, comprising the following steps: S11. dissolving a cyclodextrin derivative in a solvent, then adding resveratrol, and stirring at 40 to 80° C. for 1 to 3 hours to obtain a mixed solution; S12. The mixed solution is spray-dried to obtain the resveratrol supramolecule. 2. The method for preparing resveratrol supramolecules according to claim 1, characterized in that in step S11, the weight ratio of the cyclodextrin derivative to resveratrol and the solvent is 70-99:1-30:200-400. Most preferably, in step S11, the weight ratio of the cyclodextrin derivative to resveratrol and the solvent is 90:10:270. 3. The method for preparing resveratrol supramolecules according to claim 1, characterized in that the cyclodextrin derivative in step S11 is selected from one or more of α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxypropyl α-cyclodextrin, hydroxypropyl β-cyclodextrin, and hydroxypropyl γ-cyclodextrin. 4. The resveratrol supramolecule prepared by the preparation method according to any one of claims 1 to 3. 5. A composition, characterized in that it comprises the resveratrol supramolecule according to claim 4. 6 . The composition according to claim 5 , characterized in that it comprises the resveratrol supramolecule according to claim 4 and a Polygonum cuspidatum root extract. 7. The composition according to claim 6, characterized in that the weight ratio of resveratrol supramolecule to Polygonum cuspidatum root extract is 1 to 3:1; Most preferably, the weight ratio of resveratrol supramolecule to Polygonum cuspidatum root extract is 2:1. 8. The composition according to claim 7, characterized in that the Polygonum cuspidatum root extract is prepared by the following method: The root of Polygonum cuspidatum is extracted with a solvent to obtain an extract; the extract is concentrated and dried to obtain a solvent extract of the root of Polygonum cuspidatum; and the solvent extract of the root of Polygonum cuspidatum is obtained to obtain the root of Polygonum cuspidatum extract. 9. Use of the resveratrol supramolecule or composition according to any one of claims 4 to 8 in the preparation of products having whitening, anti-oxidation and/or acne-removing effects. 10. Use of the resveratrol supramolecule or composition according to any one of claims 4 to 8 in the preparation of cosmetics.