CN110981925B

CN110981925B - Dihydroartemisine D glucoside or salt compound thereof and application thereof in cosmetics

Info

Publication number: CN110981925B
Application number: CN201911230683.9A
Authority: CN
Inventors: 吴峰; 肖永堂; 滕景斌
Original assignee: Fuzhou Baicaotang Pharmaceutical Technology Co ltd
Current assignee: Fujian Lianke Technology Co., Ltd
Priority date: 2019-11-18
Filing date: 2019-12-05
Publication date: 2021-05-25
Anticipated expiration: 2039-12-05
Also published as: CN110981925A

Abstract

The invention relates to the technical field of cosmetics, in particular to dihydro oat alkaloid D glucoside or a salt compound thereof and application thereof in cosmetics; the dihydrooat alkaloid D glucoside and the salt compound thereof are prepared by combining dihydrooat alkaloid D with glucose and applying a biosynthesis method, the reaction has the advantages of simplicity, easiness in control, good selectivity and the like, the solubility of the dihydrooat alkaloid D glucoside and the salt compound thereof in water under the same condition is improved by more than 20 times compared with that of the corresponding dihydrooat alkaloid D, and the dihydrooat alkaloid D glucoside and the salt compound thereof are applied to biological in-vivo experiments to find that the dihydrooat alkaloid D glucoside and the salt compound thereof have the effects of resisting histamine, resisting inflammation, inhibiting allergic dermatitis, inhibiting neurodermatitis, inhibiting TRPV1, inhibiting TRPA1, relieving redness and swelling and resisting pruritus, and have very wide application prospects.

Description

Dihydroartemisine D glucoside or salt compound thereof and application thereof in cosmetics

Technical Field

The invention relates to the technical field of cosmetics, in particular to dihydro oat alkaloid D glucoside or a salt compound thereof and application thereof in cosmetics.

Background

The avenanthramide is a substance extracted from oat bran and is an alkaloid containing phenolic hydroxyl groups. The cosmetic composition has various biological activities of resisting histamine, resisting inflammation, inhibiting allergic dermatitis, inhibiting neurodermatitis, inhibiting TRPV1, inhibiting TRPA1, relieving redness and itching and the like, is a good cosmetic active ingredient, and can be widely applied to skin care such as baby products, sensitive skin care, eczema and the like. CN106511110A discloses a structural formula of dihydrooat alkaloid D.

Because the dihydrooat alkaloid has no water solubility, if the oat alkaloid and the salt thereof exist on the body surface or in the body of the animal in a glucoside form, the biological activity can be better exerted and the water solubility can be enhanced. Dihydroartemisine D-glucoside and its salt can be used as cosmetic agent with antihistaminic, antiinflammatory, allergic dermatitis inhibiting, neurodermatitis inhibiting, TRPV1 inhibiting, TRPA1 inhibiting, red swelling relieving, and pruritus resisting effects. At present, the preparation method of the glucoside compound mainly comprises a biological enzymatic synthesis method and a chemical synthesis method. The chemical synthesis method has the defects of complex synthesis process, more byproducts and the like, so the biological enzymatic synthesis method is selected for glucoside synthesis.

The existing literature data does not show the chemical structures of dihydrooat alkaloid D glucoside and salt compounds thereof and relevant reports of the application of the dihydrooat alkaloid D glucoside and the salt compounds thereof in resisting histamine, inflammation, allergic dermatitis, neurodermatitis, TRPV1, TRPA1, and relieving redness and itching.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: provides a compound with the functions of resisting histamine, resisting inflammation, inhibiting allergic dermatitis, inhibiting neurodermatitis, inhibiting TRPV1, inhibiting TRPA1, relieving redness and swelling and resisting pruritus for the skin, and researches on the application of the compound in cosmetics.

In order to solve the technical problems, the invention adopts the technical scheme that: dihydrooat alkaloid D glucoside or its salt compound has a specific structural formula (general formula 1) as follows:

wherein R is H or a salt ion comprising Na⁺、K⁺、Li⁺、Ca²⁺、Zn²⁺、Mg²⁺、NH⁴⁺Or a basic amino acid including arginine, lysine, or histidine.

Use of the compound of above 1 for the preparation of a cosmetic agent for inhibiting histamine release from mast cells.

The use of the compounds of the above general formula 1 for the preparation of anti-inflammatory cosmetic agents which are shown to inhibit the activity of nuclear factor kappa B (NF-kB).

The compound of the general formula 1 is used for preparing cosmetic agents for inhibiting allergic dermatitis.

The compound of the general formula 1 is used for preparing cosmetic agents for inhibiting neurodermatitis.

The compound of the general formula 1 is used for preparing cosmetic agents for relieving skin red swelling.

The use of the compound of the above general formula 1 for the preparation of a cosmetic agent for inhibiting capsaicin receptor (TRPV 1).

Use of the compound of the above general formula 1 for the preparation of a cosmetic agent for inhibiting transient receptor potential (TRPA 1).

The use of the compound of the above general formula 1 as an agent for the preparation of anti-itch cosmetic.

The invention also relates to a preparation method of the dihydrooat alkaloid D glucoside or the salt compound thereof, which comprises the following steps:

step 1, selecting leuconostoc mesenteroides NRRL-1299(ATCC), performing seed fermentation and shake flask culture on the strain, then centrifuging a culture solution of the leuconostoc mesenteroides, completely separating microbial cells from an enzyme liquid, and obtaining a supernatant which is a crude dextransucrase liquid;

step 2, concentrating the supernatant of the dextran sucrase obtained in the step 1, then diluting the residue with sodium acetate buffer solution, concentrating, and effectively removing the residual low molecular weight components in the enzyme-containing cell culture medium to obtain the purified dextran sucrase;

step 3, preparing 400ml of 500mM sodium acetate buffer solution, adding 500-700 g of dihydrooat alkaloid or dihydrooat alkaloid salt, 800-1200 g of cane sugar, 1500-2000 ml of DMSO, 50-100 mg of calcium chloride and 22-30U/ml of dextransucrase, fixing the volume to 1.00L with water, and reacting at 40-50 ℃ for 18-24 h under the condition of pH 5.2-5.7;

step 4, after the reaction in the step 3 is finished, adjusting the pH value of the system to be acidic, filtering to remove impurities, and collecting filtrate;

step 5, filtering the filtrate by an ultrafiltration membrane to remove enzyme in the solution, adding dried saccharomycete into the reaction solution, and fermenting at 35 ℃ for 18-24 hours to remove glucose in the reaction solution; after the reaction liquid is subjected to microfiltration and sterilization, impurities are removed through activated carbon adsorption;

and 6, purifying the reaction solution after sterilization by AB-8 resin, and performing secondary purification by silica gel to obtain the dihydrooat alkaloid D glucoside with the purity of 99% or salt compounds thereof.

The invention has the beneficial effects that: the invention combines the dihydrooat alkaloid D with glucose, prepares a dihydrooat alkaloid D glucoside and a salt compound thereof by a biosynthesis method, has the advantages of simple and easy reaction, easy control, good selectivity and the like, and has the solubility which is improved by more than 20 times than that of the corresponding dihydrooat alkaloid D under the same conditions in water; the dihydrooat alkaloid D glucoside and the salt compounds thereof are applied to biological in-vivo experiments, and found to have the effects of resisting histamine, resisting inflammation, inhibiting allergic dermatitis, inhibiting neurodermatitis, inhibiting TRPV1, inhibiting TRPA1, relieving redness and swelling and resisting pruritus, and have very wide application prospects.

Drawings

FIG. 1 is a mass spectrum of dihydrooat alkaloid D glucoside;

FIG. 2 is a nuclear magnetic resonance hydrogen spectrum of D-glucoside as a dihydrooat alkaloid;

FIG. 3 is a nuclear magnetic resonance carbon spectrum of D-glucoside as a dihydrooat alkaloid.

Detailed Description

In order to explain technical contents, achieved objects, and effects of the present invention in detail, the following description is made with reference to the accompanying drawings in combination with the embodiments.

The most key concept of the invention is as follows: the dihydrooat alkaloid D and salt thereof are combined with glucose, and an enzymatic synthesis method is adopted to prepare a dihydrooat alkaloid D glucoside and salt compound thereof, and the prepared dihydrooat alkaloid D glucoside and salt compound thereof have the effects of skin inflammation resistance, histamine resistance and the like.

The invention provides dihydro oat alkaloid D glucoside or a salt compound thereof, which has a specific structural formula (general formula 1) as follows:

The invention provides a preparation method of avenanthramide D glucoside, which comprises the following steps:

step 1, selecting leuconostoc mesenteroides NRRL-1299(ATCC), performing seed fermentation and shake flask culture on the strain, then centrifuging a culture solution of the leuconostoc mesenteroides, completely separating microbial cells from an enzyme liquid, and obtaining a supernatant which is a crude dextransucrase liquid.

And 2, concentrating the dextran sucrase supernatant obtained in the step 1, diluting the residue with 20mM sodium acetate buffer solution, concentrating, effectively removing residual low molecular weight components in the enzyme-containing cell culture medium, obtaining the purified dextran sucrase, and testing the enzyme activity to be 22-30U/ml.

Step 3, preparing 400ml of 500mM sodium acetate buffer solution, adding 500-700 g of dihydrooat alkaloid or dihydrooat alkaloid salt, 800-1200 g of cane sugar, 1500-2000 ml of DMSO, 50-100 mg of calcium chloride and 22-30U/ml of dextransucrase, fixing the volume to 10.00L with water, and reacting at 45-60 ℃ for 18-24 h under the condition of pH 5.2-5.7.

And 4, after the reaction in the step 3 is finished, adjusting the pH value of the system to be acidic, filtering to remove impurities, and collecting filtrate.

Step 5, filtering the filtrate by an ultrafiltration membrane to remove enzyme in the solution, adding dried saccharomycete into the reaction solution, and fermenting at 35 ℃ for 18-24 hours to remove glucose in the reaction solution; after the reaction liquid is subjected to microfiltration sterilization, impurities such as pigments and the like are removed through activated carbon adsorption.

And 6, purifying the reaction solution after sterilization by AB-8 resin, and performing secondary purification by using silica gel to obtain the dihydrooat alkaloid D glucoside with the purity of 99% and salt compounds thereof, wherein the mass is 110-150 g.

The Leuconostoc mesenteroides NRRL-1299 was purchased directly from ATCC.

Referring to FIGS. 1 to 3, the dihydroavenanthramide D glucoside of the present invention has the formula C₂₂H₂₅O₉N。

Nuclear magnetic hydrogen spectrum (400MHz, DMSO-d6) δ: 11.46(bs,1H),8.44(d, J ═ 8.0Hz,1H), 7.94(d, J ═ 8.0Hz,1H), 7.50(t, J ═ 12.0Hz,1H)7.14(d, J ═ 12.0Hz,2H),7.08(t, J ═ 16.0Hz, 1H),6.96(d, J ═ 12.0Hz,2H),4.64-5.27(m, 5H),3.36-3.57(m, 7H),2.86(t, J ═ 8.0Hz,2H),2.64(t, J ═ 8.0Hz,2H).

Nuclear magnetic carbon spectrum (100MHz, DMSO-d6) delta: 170.86,170.14,156.03,141.19,134.56,133.97,131.53,129.56,122.83,120.18,117.79,117.44,103.44,98.62,74.00,73.50,72.07,70.39,63.24,61.13,56.50,30.24.

High resolution Mass Spectrum (MS): calcd for C22H25O9N:447.16, found 447.16+1, (M + H)⁺)。

Example 1

The preparation method of dihydrooat alkaloid D glucoside comprises the following steps:

step 1, performing seed culture on the leuconostoc mesenteroides NRRL-1299(ATCC) strain, wherein the formula of a culture medium is as follows: 10g of sucrose, 0.17g of peptone and Na₂HPO₄0.17g of agar, 2g of agar and sterile water to 100ml of constant volume, the culture temperature is 22 ℃, the culture time is 36 hours, then the mixture is transferred into a shake flask for culture, and the formula of a shake flask culture medium is as follows: sucrose 5g, peptone 0.2g, Na₂HPO₄0.17g, the volume of the sterile water is 100ml, the culture temperature is 22 ℃, and the culture time is 36 h. And then centrifuging the culture solution of the leuconostoc mesenteroides, completely separating the microbial cells from the enzyme liquid, and obtaining a supernatant which is crude enzyme solution of dextran sucrase.

And 2, concentrating the dextran sucrase supernatant by 4 times (the molecular weight cutoff value is 100kDa), then diluting the residue by 4 times with 20mM sodium acetate buffer solution, concentrating by 4 times, effectively removing residual low molecular weight components in the enzyme-containing cell culture medium to obtain the purified dextran sucrase, and testing the enzyme activity to be 22U/ml.

Step 3, preparing 400ml of 500mM sodium acetate buffer solution, weighing 27.2g of anhydrous sodium acetate, diluting with water to a constant volume of 400ml, then adding 500g of dihydrooat alkaloid, 800g of cane sugar, 1500 DMSO (1500 ml), 50mg of calcium chloride and 22U/ml of dextran sucrase, diluting with water to a constant volume of 10.00L, adjusting the pH value of the system to 5.2, adjusting the temperature to 40 ℃, and reacting for 18 h.

And 4, adjusting the pH value of the system to 2.5, filtering to remove precipitates, and collecting filtrate.

Step 5, filtering the filtrate by an ultrafiltration membrane to remove enzyme in the solution, adding dried yeast (active yeast, purchased from Angel corporation) into the reaction liquid, fermenting at 35 ℃ for 18h to remove glucose in the reaction liquid, wherein the mass volume percentage of the yeast in the reaction liquid is 1% (mass g of the yeast/volume ml of the reaction liquid); after the reaction liquid is subjected to microfiltration sterilization, impurities such as pigments and the like are removed through activated carbon adsorption.

And 6, carrying out primary purification on the reaction liquid after sterilization through AB-8 resin, eluting AB-8 with absolute ethyl alcohol, concentrating the eluent until the eluent is completely dried to obtain a 90% purity dihydroavenanthramide D glucoside crude product, and then carrying out secondary purification on the crude product through a silica gel column to obtain the 99% purity dihydroavenanthramide D glucoside, wherein R in the general formula 1 is H, and the mass of R is 114 g.

The solubility test results of dihydrooat alkaloid D glucoside and dihydrooat alkaloid D are as follows: dihydroavenanthramide D is practically insoluble in water at 25 ℃ and has a solubility of less than 0.01g/L, and dihydroavenanthramide D glucoside has a solubility of more than 0.4g/L in water at 25 ℃.

Example 2

The preparation method of the dihydrooat alkaloid D sylvite glucoside comprises the following steps:

step 1, performing seed culture on the leuconostoc mesenteroides NRRL-1299(ATCC) strain, wherein the formula of a culture medium is as follows: 10g of sucrose, 0.17g of peptone and Na₂HPO₄0.17g of agar, 2g of agar and sterile water to 100ml of constant volume, the culture temperature is 24 ℃, the culture time is 42 hours, then the mixture is transferred into a shake flask for culture, and the formula of a shake flask culture medium is as follows: sucrose 5g, peptone 0.2g, Na₂HPO₄0.17g, the volume of the sterile water is 100ml, the culture temperature is 24 ℃, and the culture time is 42 h. Centrifuging the culture solution of Leuconostoc mesenteroides, and completely separating microbial cells from enzyme liquid to obtainThe obtained supernatant is crude enzyme solution of dextran sucrase.

And 2, concentrating the dextran sucrase supernatant by 6 times (the molecular weight cutoff value is 100kDa), then diluting the residue by 4 times with 20mM sodium acetate buffer solution, concentrating by 4 times, effectively removing residual low molecular weight components in the enzyme-containing cell culture medium to obtain the purified dextran sucrase, and testing the enzyme activity to be 26U/ml.

Step 3, preparing 400ml of 500mM sodium acetate buffer solution, weighing 27.2g of anhydrous sodium acetate, diluting with water to a constant volume of 400ml, adding dihydrooat alkaloid D600g, sucrose 1000g, DMSO1700ml, calcium chloride 70mg and dextran sucrase (26U/ml), diluting with water to a constant volume of 10.00L, adjusting the pH value of the system to 5.4, adjusting the temperature to 45 ℃, and reacting for 22 h.

And 4, adjusting the pH value of the system to 2.7, filtering to remove precipitates, and collecting filtrate.

Step 5, filtering the filtrate by an ultrafiltration membrane to remove enzyme in the solution, adding dried yeast (active yeast, purchased from Angel corporation) into the reaction liquid, fermenting for 22h at 35 ℃ to remove glucose in the reaction liquid, wherein the mass volume percentage of the yeast in the reaction liquid is 1.2% (mass g of the yeast/volume ml of the reaction liquid); after the reaction liquid is subjected to microfiltration sterilization, impurities such as pigments and the like are removed through activated carbon adsorption.

And 6, carrying out primary purification on the reaction liquid after sterilization through AB-8 resin, eluting the AB-8 resin through absolute ethyl alcohol, concentrating eluent until the eluent is completely dried to obtain a 90% purity dihydroavenanthramide D glucoside crude product, and then carrying out secondary purification on the crude product through a silica gel column to obtain the 99% purity dihydroavenanthramide D glucoside, wherein the mass of the dihydroavenanthramide D glucoside is 135 g.

Step 7, dissolving 0.05mol of dihydrooat alkaloid D glucoside in absolute ethyl alcohol, heating to 40 ℃, introducing nitrogen for protection, dropwise adding 10% potassium hydroxide solution until complete neutralization, stirring for neutralization reaction for 30 minutes, and then rotatably evaporating ethanol and water to dryness to obtain a dihydrooat alkaloid D sylvite glucoside product, namely a product with R as K in the general formula 1⁺。

Example 3

Experiment for inhibiting substance P from causing mast cells to release histamine

Culturing in vitro mast cells; adding substance P with the same concentration and dihydrooat alkaloid D glucoside (0.1%, 0.5%, 1%) with different concentrations into different culture media for culturing; and finally, comparing the release amount of histamine in the solution, and observing the inhibition effect of the dihydrooat alkaloid D glucoside on histamine release. The experiment was set with both blank and control groups: the culture medium is added with substance P with the same concentration, and dihydrooat alkaloid D glucoside is not added. The experimental results are shown in table 1;

TABLE 1

The results show that: the dihydrooat alkaloid D glucoside has an inhibitory effect on histamine released by mast cells, and the release amount of histamine is reduced by over 80%.

Example 4

Inhibition assay of Dihydroartemisine D glucoside on NF-kB Activity

NF-kB is a regulator protein factor for many inflammatory processes. In most cells, NF-kB is in the cytoplasm and binds the suppressor protein, IkB. When stimulated by extracellular stimulating factors, such as TNF- α, the cells result in the activation of IkB kinase, which leads to phosphorylation, generalization, and degradation of the IkB protein. Once IkB is degraded and NF-kB protein subunits are free, the transcription gene is opened, a large amount of expression is realized, and inflammation is continuously generated. Human keratinocytes were seeded in 96-well plates and transfected with 0.1. mu.g of pNF-kB-Luc reporter plasmid using Lipofectamine 2000 kit. Cells were treated with 5, 10 and 20mg/L dihydroavenanthramide D glucoside for 24 hours and then induced with 100. mu.g/ml TNF-. alpha.for 24 hours. Only 100. mu.g/ml TNF-. alpha.induction was used as a blank, and only 10mg/L dihydroavenanthramide D-glucoside treatment was used as a control. After induction, luciferase readings were performed using the Promega kit to observe the inhibition according to the manufacturer's instructions, see tables 2 and 3.

TABLE 2

Serial number	Inhibiting effect	Identification criteria
				1	++++	80 to 100 percent inhibition
2	+++	60 to 80 percent of inhibition
			3	++	40-60% inhibition
4	+	20 to 40 percent of inhibition
			4	-	Without inhibition

TABLE 3

In keratinocytes of the blank and dihydroavenanthramide D-glucoside treated groups, we observed that dihydroavenanthramide D-glucoside could inhibit the degradation of IkB-alpha protein. From Table 2, it can be seen that different concentrations of D-glucoside, which is a dihydroavenanthramide, have different inhibitory effects on IkB-alpha degradation, and the inhibitory effect of D-glucoside, which is a dihydroavenanthramide, is most significant at a concentration of 20 mg/L. In the blank control group without addition of avenanthramide D, we observed that IkB-alpha protein was degraded in TNF-alpha induced keratinocytes, thereby inhibiting NF-kB activity. However, in the experimental group to which dihydroavenanthramide D-glucoside (10mg/L concentration) was added, no degradation of IkB-alpha protein was observed, and NF-kB activity was not inhibited.

Example 5

Dihydroartemisine D potassium glucoside inhibition of allergic dermatitis test

Female mice (Balb/c) of 8-10 weeks were selected, 0.01% Tetradecanoyl Phorbol Acetate (TPA) was used as an inducer for allergic edema, different concentrations of dihydroavenanthramide D potassium glucoside (0.1%, 0.5%, 1%) were prepared as experimental groups, and TPA and dihydroavenanthramide D potassium glucoside were prepared with 100% ethanol. Applying 10 microliters of TPA to the right ear of a mouse, applying 5 mice as a group, applying ethanol only as a blank group, applying TPA only as a control group, observing the allergic condition after 6 hours, and then performing parallel experiments, wherein the red swelling part of the allergic dermatitis of the mouse is treated by dihydrooat alkaloid D potassium salt glucoside with the concentration of 0.1%, 0.5% and 1%, respectively, the level of the inhibition of the allergy can be confirmed by measuring the thickness of the ear, and the inhibition rate of the allergic dermatitis of the ear after ethanol treatment is expressed in percentage, please refer to Table 4.

TABLE 4

Administration of drugs	Inhibition ratio of allergic dermatitis%
		Blank group(ethanol)	Has no allergy
Control group (TPA)	0
		TPA (0.01%) + Dihydroantia D potassium glucoside (0.1%)	28
TPA (0.01%) + Dihydroantia D potassium glucoside (0.5%)	46
		TPA (0.01%) + Dihydroantia D potassium glucoside (1%)	68

As can be seen from the above table: different concentrations of dihydrooat alkaloid D potassium glucoside have inhibitory effect on allergic dermatitis, and 1% of dihydrooat alkaloid D potassium glucoside has remarkable effect on allergic dermatitis.

Example 6

Dihydroavenanthramide D sylvite glucoside inhibition of neurodermatitis test, see table 5;

TABLE 5

Selection of patients: 80 patients with neurodermatitis were selected, and the average age was 30-35 years. 30 patients with neurodermatitis were randomly divided into 4 groups of 20 persons each, each group being rubbed with the control group, and composition 1 to composition 3. Symptoms of the patient prior to treatment: the common symptoms of 60 patients with neurodermatitis are severe itching and slight pain of affected parts, local occurrence of round or polygonal flat papules with the size of millet granular mung beans, skin color, light red or light brown, dryness, fine crumbling and desquamation, and the early stage of neurodermatitis.

The amount of the wiping: the ointment for wiping is applied to the affected part 3 times each day, respectively in the morning, the middle and the evening, and the affected part is cleaned with warm water before each wiping, with 7 days as a treatment course, and is continuously wiped for three treatment courses; the comparative effect after three treatment sessions is shown in table 6:

TABLE 6

As can be seen from the above table: the composition 1, the composition 2 and the composition 3 all contain dihydrooat alkaloid D sylvite glucoside active components and have obvious effect on inhibiting neurodermatitis.

Example 7

Test method for relieving skin redness and swelling:

to test the anti-redness effect of the invention, the invention was formulated into various concentrations of dihydroavenanthramide, D-potassium glucoside, compositions and tested on human skin, see Table 7.

TABLE 7

The red swelling removal test method is as follows: 30 volunteers aged 20-50 years without prior history of allergy and systemic anaphylaxis were selected, and with consent, a 0.2% sodium lauryl sulfate solution was applied to the forearm of the subject to induce redness of the skin, and the area was protected with a closed patch for 24 hours. After 24 hours, the patch is removed and the stimulus-induced redness can be assessed using the a-value of Minolta Chroma Meter and archived by photography. Immediately after the measurement, the area was treated with the composition of the present invention. The using method comprises the following steps: the medicine is continuously used for 14 days, once in the morning and at night, and is taken and archived. The photographs on day 1 and day 14 were compared and the results were observed.

Comparative group 1: the forearm of the 30 subjects was also treated by the same method as described above. After the assay, the area was treated with placebo. The using method comprises the following steps: the medicine is continuously used for 14 days, once in the morning and at night, and is taken and archived. The photographs on day 1 and day 14 were compared and the results were observed.

Comparative group 2: the forearm of the 30 subjects was also treated by the same method as described above. After the assay, no drug treatment was used. The using method comprises the following steps: the medicine is continuously used for 14 days, once in the morning and at night, and is taken and archived. The photographs on day 1 and day 14 were compared, and the results were observed, and the criteria were as shown in Table 8.

TABLE 8

Serial number	Effect	Identification criteria
				1	+ significant effect	The red swelling can be eliminated by over 75 percent
2	+ effective	The red swelling can be eliminated by more than 50%
			3	-invalidation	The red swelling is eliminated by less than 10%

The test results are in Table 9;

TABLE 9

	Marked effect +	Is effective	Ineffectiveness
				Composition
1	10	14	6
				Composition 2	17	8	5
Composition 3	22	5	3
				Comparative group 1	0	5	25
Comparative group 2	0	3	27

As can be seen from the above table: the composition 1, the composition 2 and the composition 3 all contain dihydrooat alkaloid D sylvite glucoside active components and have obvious treatment effect on skin red swelling. Different concentrations of dihydrooat alkaloid D potassium glucoside are added into the

compositions

1, 2 and 3 respectively, but the effect of the composition 3 is more obvious as shown in the table, so that the addition amount of the dihydrooat alkaloid D potassium glucoside with the concentration of 1% has obvious effect of relieving the skin redness and swelling.

Example 8

TRPV1 antagonistic activity assay:

TRPV1 is one of the trp (transient Receptor potential) family of transient Receptor potential receptors, a nociceptor that is specifically activated by capsaicin, nociceptive thermal stimulation, and acidification. The experiment evaluates the inhibition of dihydrooat alkaloid D glucoside on capsaicin-induced substance P. Test protocol, blank group, control group and test group, blank group (neuron cell and normal saline), control group (neuron cell and capsaicin), test group (control group added with dihydrooat alkaloid D glucoside with specified concentration). Thus, the concentrations of the dihydroavenanthramide D glucoside solutions in this test were 0.01%, respectively; 0.1 percent; 0.5%, the incubation time was 10 minutes. After 10 minutes of cell co-culture, culture supernatants were taken and substance P was assessed from the culture supernatants by ELISA analysis.

Test result data table 10;

watch 10

From table 10, it can be seen that dihydroavenanthramide D glucoside has good inhibitory effect on capsaicin-induced substance P and good antagonistic effect on TRPV 1.

Example 9

TRPA1 inhibitory activity test:

the experiment is to observe that dihydrooat alkaloid D glucoside has inhibition on TRPV1 of keratinocytes. The test scheme comprises setting a blank group, a control group and a test group, wherein the blank group is used for recording the calcium ion potential of the keratinocyte after being excited by TRPA1 agonist (2 mu M polygodial), the test group is used for adding 0.1%, 0.5% and 1% dihydrooat alkaloid D glucoside after being excited by TRPA1 agonist (2 mu M polygodial), and the calcium ion potential of the keratinocyte is recorded after 5 minutes. After the keratinocyte and a calcium ion fluorescent probe Fura-2 are co-cultured for 2 hours, a calcium ion imaging system is used for detection.

The test results are shown in Table 11:

TABLE 11

From table 11, the calcium ion potential was maximally shifted after the blank group of keratinocytes were challenged with TRPA1 agonist. Different concentrations of dihydrooat alkaloid D glucoside are added to inhibit TRPA1 of keratinocytes, and 1% of dihydrooat alkaloid D glucoside has the best inhibition effect on TRPA 1.

Example 10

Dihydrooat alkaloid D glucoside anti-itch Activity test:

the hair on the nape of the neck, approximately 1cm by 1cm, was shaved the day before the experiment using 8-10 week male C57BL/6 mice. The test samples were divided into blank control group, model group and experimental group (5, 10 and 20mg/L), and each group had 5 mice. Injecting 50 mul of vehicle solvent into the unhairing positions of the backs and the necks of mice in a blank control group and a model group by using an injection needle, and injecting 50 mul of dihydrooat alkaloid D glucoside solution with different concentrations into an experimental group; after 30 minutes, 50. mu.l of physiological saline was injected into the blank control group, and 50. mu.l of histamine solution (100. mu.M) was injected into the remaining groups, and video-recorded by a video camera for 30 minutes. After the experiment is finished, the video is played back to carry out hind limb scratching behavior statistics, the number of times of injecting the hind limb scratching injection part on the same side by the mouse in the observation period is counted by one time of scratching of the mouse from the hind limb lifting scratching injection part to the putting down or licking of the mouse, and the table 12 is referred.

TABLE 12

The results show that the scratching times of the mice after the histamine injection in the model group are 45.7 +/-5.1 times, and are obviously increased compared with the blank control group (3.2 +/-1.6 times). The number of scratching of mice in the 5mg/L dose group of the dihydroavenanthramide D glucoside is 27.2 +/-7.5, the number of scratching of mice in the 10mg/L dose group is 17.9 +/-3.5, and the number of scratching of mice in the 20mg/L dose group is 8.3 +/-2.2.

In summary, the invention combines the active ingredients of the oat, namely the dihydrooat alkaloid D and the glucose, and the hydroxyl of the dihydrooat alkaloid D and the hydroxyl of the glucose are etherified to synthesize the dihydrooat alkaloid D glucoside, so that the reaction has the advantages of simplicity, easiness in control, good selectivity and the like, and the solubility of the dihydrooat alkaloid D in water under the same condition is improved by more than 20 times than that of the corresponding dihydrooat alkaloid D; the dihydrooat alkaloid D glucoside and the salt compounds thereof are applied to biological in-vivo experiments, and found to have the effects of resisting histamine, resisting inflammation, inhibiting allergic dermatitis, inhibiting neurodermatitis, inhibiting TRPV1, inhibiting TRPA1, relieving redness and swelling, resisting pruritus and the like, and have very wide application prospects.

The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all equivalent changes made by using the contents of the present specification and the drawings, or applied directly or indirectly to the related technical fields, are included in the scope of the present invention.

Claims

1. A compound of formula 1, characterized by the structural formula:

wherein R is H or salt ion, and the salt ion is Na⁺、K⁺、Li⁺、Ca²⁺、Zn²⁺、Mg²⁺、NH⁴⁺Or a basic amino acid which is arginine, lysine or histidine.

2. Use of a compound of formula 1 according to claim 1 for the preparation of a cosmetic agent for inhibiting histamine release from mast cells.

3. The use of the compound of formula 1 according to claim 1 for the preparation of an anti-inflammatory cosmetic agent manifested in the inhibition of nuclear factor kappa B activity.

4. Use of the compound of formula 1 according to claim 1 as a cosmetic agent for the preparation of an allergic dermatitis inhibition.

5. Use of the compound of formula 1 according to claim 1 for the preparation of a cosmetic agent for the inhibition of neurodermatitis.

6. Use of a compound of formula 1 according to claim 1 for the preparation of a cosmetic agent for relieving skin redness and swelling.

7. Use of a compound of formula 1 according to claim 1 as a cosmetic agent for the preparation of a TRPV1 inhibiting agent.

8. Use of a compound of formula 1 according to claim 1 as a cosmetic agent for the preparation of a TRPA1 inhibiting agent.

9. Use of a compound of formula 1 according to claim 1 as an agent for the preparation of anti-pruritic cosmetic agents.

10. A process for the preparation of a compound of formula 1 according to claim 1, comprising the steps of:

step 1, selecting leuconostoc mesenteroides NRRL-1299, performing seed fermentation and shake flask culture on the strain, then centrifuging a culture solution of the leuconostoc mesenteroides, completely separating microbial cells from an enzyme liquid, and obtaining a supernatant which is a crude enzyme solution of dextran sucrase;

step 3, preparing 400ml of 500mM sodium acetate buffer solution, adding 500-700 g of dihydrooat alkaloid D or dihydrooat alkaloid D salt, 800-1200 g of cane sugar, 1500-2000 ml of DMSO, 50-100 mg of calcium chloride and 22-30U/ml of dextransucrase, fixing the volume to 10.00L with water, and reacting at 40-50 ℃ for 18-24 h under the condition of pH 5.2-5.7;

and 6, purifying the reaction solution after sterilization by AB-8 resin, and performing secondary purification by using silica gel to obtain the compound shown in the general formula 1.