CN113318036A

CN113318036A - New application of artemisia plant extract water

Info

Publication number: CN113318036A
Application number: CN202110744889.4A
Authority: CN
Inventors: 卢永杰; 程路诗; 张炽坚; 张兵; 江桦; 关焕敏; 艾勇; 何廷刚
Original assignee: Hua An Tang Biotech Group Co ltd
Current assignee: Hua An Tang Biotech Group Co ltd
Priority date: 2021-06-30
Filing date: 2021-06-30
Publication date: 2021-08-31

Abstract

The invention relates to the technical field of plant extracts, in particular to application of artemisia argyi plant extract water in preparation of moisturizing liposomes. The artemisia argyi plant extract water is used as a solvent for preparing the liposome, and the obtained liposome is unexpectedly found to have natural preservative and moisturizing effects, so that the liposome is preserved without adding a preservative additionally, and the liposome can be applied to cosmetics to improve the moisturizing performance of the cosmetics.

Description

New application of artemisia plant extract water

Technical Field

The invention relates to the technical field of plant extracts, in particular to application of artemisia argyi plant extract water in preparation of moisturizing liposomes.

Background

Blumea balsamifera (Blumea balsamifera DC) belongs to perennial woody herbaceous plants or subshrubes, and is also called Blumea balsamifera, Blumea balsamifera and the like. The main medicinal parts are whole plants or aboveground parts, are produced in Guizhou, Guangxi, Guangdong, Hainan and other places, and are important medicinal materials of Chinese linationalities, Zhuang nationality, Miao nationality and other minority nationalities. The whole herb contains volatile components, the main functional component is L-borneol, and has better biological activities of diminishing inflammation, easing pain, resisting bacteria, resisting oxidation, resisting virus and the like, the extracts of the L-borneol, the Artemisia powder, the Artemisia tablet and the blumea balsamifera oil are important Chinese patent medicine raw material medicines, and more than 30 Chinese patent medicines taking the Artemisia powder and the Artemisia tablet as raw materials, such as Yanglishuang dropping pills, heart and stomach pain relieving capsules, Jinhoujianjian spray, Yinxiao acne tincture, compound herba Solidaginis spray, Wanjin fragrant fog, Yankang buccal tablets and the like. Besides, the blumea balsamifera oil is widely applied to the fields of chemical industry, spices, essence and the like due to the unique fragrance and better biological activity. The folium Artemisiae Argyi powder is a powdery crude extract obtained by steam distilling folium Artemisiae Argyi and twig, contains volatile oil, main ingredient of L-borneol, and small amount of essential oil, L-Camphora and sesquiterpene. The blumea balsamifera is determined by various chemical components to have rich biological activity, strong antibacterial activity, penetration promoting effect and antioxidant activity, and the functions of relieving swelling and pain, smoothing nasal orifices, dispelling wind and removing dampness, warming spleen and stomach and promoting blood circulation and the like. Researches show that the blumea balsamifera has photosensitivity, inhibitory effect on escherichia coli and candida albicans and other biological activities. In recent years, blumea balsamifera has been paid more and more attention by researchers due to its remarkable biological activity, and the research contents mainly focus on the aspects of chemical components, biological activity, development and application, and the like. At present, the application of blumea balsamifera extract water in liposome is not reported.

Disclosure of Invention

In view of the above, the invention provides an application of the artemisia argyi plant extract water in preparing the moisturizing liposome, and the artemisia argyi plant extract water is applied in the liposome, so that the liposome has moisturizing and preservative effects.

The specific technical scheme is as follows:

the invention provides application of artemisia argyi plant extract water in preparation of moisturizing liposomes.

The invention uses the artemisia plant extract water as the solvent of the moisturizing liposome, so that the obtained liposome has natural preservative and moisturizing effects.

In the invention, the blumea balsamifera plant extract water is obtained by distilling blumea balsamifera leaves under a low-temperature vacuum condition. The distillation temperature is 30-50 ℃, the pressure is-70 to-90 kPa, and the time is 5-8 h.

In the invention, the preparation of the moisturizing liposome comprises the following steps:

step 1: mixing phospholipid, cholesterol, medicine and folium Artemisiae Argyi plant water to form suspension; the drug comprises a fat-soluble drug;

step 2: and shearing the suspension at a high speed under the heating and heat preservation state to obtain the liposome emulsion.

The invention mixes phospholipid, insoluble medicine and folium Artemisiae Argyi plant extract water to obtain suspension, and disperses at high speed by shearing at proper temperature, and the phospholipid and insoluble medicine are pulverized to form micrometer scale. Under the micron scale, after long-time shearing, the insoluble drug particles and the phospholipid particles interact to emulsify, and then self-assemble to form the micron-scale liposome.

The method does not use toxic and harmful organic solvents in the process of preparing the fat-soluble drug liposome, and has safe and environment-friendly production process and high product safety; the preparation equipment and the process are simple, and the method is suitable for large-scale production and improves the production efficiency.

In step 1 of the present invention, the fat-soluble drug comprises: one or more of glabridin, ceramide, resveratrol and fat-soluble vitamins, preferably glabridin. The invention finds that the moisturizing effect of the liposome can be synergistically enhanced by compounding the glabridin and the sodium nervonate extract water.

The phospholipid comprises one or more of soybean lecithin, egg yolk lecithin, hydrogenated lecithin, dilauroyl phosphatidylcholine, dimyristoyl phosphorylcholine, dipalmitoyl phosphorylcholine, distearoyl phosphatidylcholine, dioleoyl phosphatidylcholine, dilauroyl phosphatidylethanolamine, dimyristoyl phosphatidylethanolamine, dipalmitoyl phosphatidylethanolamine, distearoyl phosphatidylethanolamine and dioleoyl phosphatidylethanolamine.

The medicament further comprises: a water-soluble drug. The water-soluble medicine comprises one or more than two of sodium hyaluronate, nano metal colloid dispersion liquid and water-soluble vitamins.

Before mixing, the method also comprises the following steps: adding a surfactant. The addition of a surfactant can facilitate the solubilization of the fat-soluble drug.

The surfactant is ionic surfactant and/or nonionic surfactant, wherein the cationic surfactant comprises quaternary ammonium salt, the anionic surfactant comprises higher fatty acid salt or sulfate salt, and the nonionic surfactant comprises one or more than two of tween, span, PEG fatty acid ester and polyglycerol-10-hard fatty acid ester.

Before mixing, the method also comprises the following steps: the polyol is added. The addition of the polyol can further facilitate the dissolution of a portion of the fat-soluble drug. Glabridin has low solubility in polyhydric alcohol.

The polyhydric alcohol comprises one or more of glycerol, propylene glycol, butylene glycol, pentylene glycol, and isoprene glycol.

In the invention, according to the mass percentage,

2-15% of phospholipid;

0.01-5% of a medicament;

0-5% of cholesterol;

the balance being water.

Preferably, the amount of the surfactant is, in mass percent,

2-10% of phospholipid;

0.01-4% of a medicament;

0-4% of cholesterol;

the balance being water.

More preferably, the amount of the surfactant is, in mass percent,

2-8% of phospholipid;

0.1-3% of a medicament;

0-3% of cholesterol;

when the medicament is a fat-soluble medicament and a water-soluble medicament, the dosage ratio of the fat-soluble medicament and the water-soluble medicament is not particularly limited.

In the invention, the weight percentage of the surfactant is 0.1-20%, preferably 0.1-15%, and more preferably 0.1-8%; 5 to 60% of a polyol, preferably 5 to 45%, more preferably 5 to 30%.

In step 2 of the invention, the heating temperature of the suspension liquid needs to be higher than the phase transition temperature of the phospholipid. The heating temperature is 50-100 ℃, and preferably 50-70 ℃; then, maintaining the heating temperature for shearing, wherein a shearing disperser is adopted for shearing; the shearing rate is 8000-20000 rpm (rotor diameter is 13mm), the shearing time is more than 30min, preferably more than 60min at the shearing rate of 8000-15000 rpm (rotor diameter is 13mm), more preferably 60-90 min. The liposome obtained after shearing is micron-sized.

The nano-scale liposome has better dispersibility and stability and better skin permeability. Therefore, after the high-speed shearing is performed in step 2 of the present invention, the method further comprises: homogenizing under high pressure. The high-pressure homogenization can crush and thin the micron-sized liposome to obtain the nano-sized liposome. The high-pressure homogenization specifically comprises the following steps: each cycle was carried out at a pressure of 400bar, 600bar, 800bar, 1000bar for 2 or more times.

The invention also provides a liposome which comprises medicine, phospholipid, cholesterol and the water extracted from the plants of the Aina.

The dosage of each component of the liposome provided by the invention is the same as that of each component of the raw materials prepared in the technical scheme of the application of the artemisia plant extract water in the preparation of the moisturizing liposome, and the details are not repeated herein.

According to the technical scheme, the invention has the following advantages:

the invention provides application of artemisia argyi plant extract water in preparation of liposome. The artemisia argyi plant extract water is used as a solvent for preparing the liposome, and the obtained liposome is unexpectedly found to have natural preservative and moisturizing effects, so that the liposome is preserved without adding a preservative additionally, and the liposome can be applied to cosmetics to improve the moisturizing performance of the cosmetics.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without inventive exercise.

FIG. 1 is an optical microscope photograph (3 μm on scale) of liposomes in an emulsion prepared in step 1 of example 2 of the present invention;

FIG. 2 is an optical microscope photograph (3 μm on scale) of liposomes in the emulsion obtained in step 1 of example 3 of the present invention;

FIG. 3 is an optical microscope photograph (3 μm on scale) of liposomes in the emulsion obtained in step 1 of example 4 of the present invention;

FIG. 4 is an optical microscope photograph (3 μm on scale) of liposomes in the emulsion obtained in step 1 of example 5 of the present invention;

FIG. 5 is an optical microscope photograph (3 μm on scale) of liposomes in the emulsion obtained in step 1 of example 6 of the present invention;

FIG. 6 is an optical microscope photograph (3 μm on scale) of liposomes in an emulsion prepared in step 1 of example 7 of the present invention;

FIG. 7 is an optical microscope photograph (3 μm on scale) of liposomes in an emulsion prepared in step 1 of example 9 of the present invention;

FIG. 8 is a bar graph showing the expression levels of AQP3 protein provided in test example 3 of the present invention;

FIG. 9 is a histogram showing the expression level of Claudin1 protein provided in test example 3 of the present invention.

Detailed Description

In order to make the objects, features and advantages of the present invention more obvious and understandable, the technical solutions in the embodiments of the present invention will be clearly and completely described below, and it should be apparent that the embodiments described below are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The reagents in the examples of the present invention are all commercially available.

Wherein, the high-speed homogeneous dispersion machine of experimental type: IKA, Germany.

Experimental high-pressure homogenizer: suzhou ATS Inc.

Zeta nanometer particle size analyzer: malvern, uk.

Reagent:

soybean lecithin: PC > 95%, Sigma-Aldrich Sigma Aldrich trade company, Shanghai.

Glabridin: HPLC purity is more than or equal to 90 percent, Qinghai lake pharmaceutical industry Co.

Cholesterol: purity > 96%, Sigma-Aldrich Sigma Aldrich trade ltd.

Polyglycerol-10-stearate: heliochemical trade (shanghai) limited.

Butanediol: purity > 98%, manufactured by Nippon Daiillo Co., Ltd.

Example 1

This example is a preparation of an artemisia plant extract water, which comprises the following specific steps:

selecting fresh blumea balsamifera leaves as raw materials, adding no external water and solvent, and distilling water and other volatile substances in the plant under low-temperature vacuum condition (extraction temperature of 35 ℃, extraction pressure of-90 kPa and extraction time of 6h) to obtain the water extracted from the blumea balsamifera plant.

Example 2

The preparation formula of glabridin liposome in this example is shown in table 1, and the specific preparation steps are as follows:

(1) adding phospholipid (PC 95%), glabridin and cholesterol into the prescribed amount of sodium nerved water, heating in water bath at 50 deg.C, and high-speed shearing with experimental IKA high-speed shearing disperser at 10000rpm for 30min to obtain uniform emulsion.

(2) And (2) homogenizing the uniform emulsion obtained in the step (1) under high pressure, and circulating for 2 times under 400/600/800/1000bar pressure respectively to obtain uniform glabridin liposome.

As can be seen in FIG. 1, after high shear in step 1, a multi-lamellar globular structure resembling liposomes is observed in the emulsion in step 1.

Examples 3 to 5

(1) adding phospholipid (PC 95%), glabridin and cholesterol into the prescribed amount of sodium nerved water, heating in water bath at 60 deg.C, and high-speed shearing with experimental IKA high-speed shearing disperser at 10000rpm for 60min to obtain uniform emulsion.

As can be seen from FIGS. 2 to 4, after the high-speed shearing in step 1, a multi-layer spherical structure similar to liposome can be observed in the emulsion in step 1.

Example 6

(1) adding phospholipid (PC 95%), polyglycerol-10-stearate, glabridin and cholesterol into the total amount of folium Artemisiae Argyi plant extract water, heating in water bath at 50 deg.C, and high-speed shearing with an experimental IKA high-speed shearing disperser at 10000rpm for 30min to obtain uniform emulsion.

As can be seen in FIG. 5, after high shear in step 1, a multi-lamellar globular structure resembling liposomes is observed in the emulsion in step 1.

Examples 7 to 8

(1) adding phospholipid (PC 95%), polyglycerol-10-stearate, glabridin and cholesterol into the mixture of the sodium-extracted water and butanediol, heating in a water bath at 50 deg.C, and high-speed shearing for 30min at 10000rpm by using an experimental IKA high-speed shearing disperser to obtain uniform emulsion.

As can be seen in FIG. 6, in example 7, after high shear in step 1, a multi-lamellar globular structure resembling liposomes is observed in the emulsion in step 1.

Example 9

(1) adding phospholipid (PC 95%), polyglycerol-10-stearate, glabridin and cholesterol into the prescribed amount of sodium neratide extraction water, heating in water bath at 60 deg.C, and high-speed shearing with an experimental IKA high-speed shearing disperser at 10000rpm for 60min to obtain uniform emulsion.

Example 10

(1) adding phospholipid (PC 95%), polyglycerol-10-stearate, glabridin and cholesterol into the mixture of the folium Artemisiae Argyi plant extract water and butanediol, heating in water bath at 50 deg.C, and high-speed shearing with an experimental IKA high-speed shearing disperser at 8000rpm for 30min to obtain uniform emulsion.

(2) And (2) homogenizing the uniform emulsion obtained in the step (1) under high pressure, and circulating for 2 times under 400/600/800/1000bar pressure to obtain transparent and uniform glabridin liposome.

Example 11

(1) adding phospholipid (PC 95%), polyglycerol-10-stearate, glabridin and cholesterol into the mixture of the folium Artemisiae Argyi plant extract water and butanediol, heating in water bath at 60 deg.C, and high-speed shearing with an experimental IKA high-speed shearing disperser at 10000rpm for 30min to obtain uniform emulsion.

Example 12

(1) adding phospholipid (PC 95%), polyglycerol-10-stearate, glabridin and cholesterol into the mixture of the folium Artemisiae Argyi plant extract water and butanediol, heating in water bath at 50 deg.C, and high-speed shearing with an experimental IKA high-speed shearing disperser at 10000rpm for 60min to obtain uniform emulsion.

Example 13

(1) adding phospholipid (PC 95%), polyglycerol-10-stearate, glabridin and cholesterol into the mixture of the folium Artemisiae Argyi plant extract water and butanediol, heating in water bath at 50 deg.C, and high-speed shearing with an experimental IKA high-speed shearing disperser at 10000rpm for 90min to obtain uniform emulsion.

Comparative example 1

(1) adding phospholipid (PC 95%), glabridin and cholesterol into water, heating in water bath at 60 deg.C, and high-speed shearing with experiment type IKA high-speed shearing disperser at 10000rpm for 60min to obtain uniform emulsion.

Comparative examples 2 to 3

The comparative example is the preparation of glabridin liposome, the formula is shown in table 1, and the preparation steps are as follows:

(1) dissolving phospholipid (PC 95%), polyglycerol-10-stearate, glabridin and cholesterol in 200g methanol-chloroform (mass ratio of 62:38) to obtain clear and transparent solution, and performing rotary evaporation to remove organic solvent to obtain the film.

(2) And (2) placing the film obtained in the step (1) in a water bath at 60 ℃, and adding a mixed solution of the prescribed amount of the artemisia plant extraction water and the butanediol while stirring to obtain a crude emulsion. Homogenizing the crude emulsion under high pressure, and circulating for 2 times under 400/600/800/1000bar pressure to obtain glabridin liposome.

Comparative example 4

(1) adding phospholipid (PC 95%), polyglycerol-10-stearate, glabridin and cholesterol into a mixture of water and butanediol, heating in water bath at 30 deg.C, and high-speed shearing with an experimental IKA high-speed shearing disperser at 10000rpm for 30min to obtain uniform emulsion.

TABLE 1 example and comparative example formulations

Test example 1

The glabridin liposomes prepared in example 9 and comparative example 1 were dispersed using PBS as a dispersant, and the average particle diameter was measured using a Zeta sizer; separating the encapsulated and free glabridin by an ultrafiltration centrifugal tube, detecting the glabridin content by HPLC and calculating the encapsulation rate. Test results are shown in table 2.

As is clear from the results in Table 2, glabridin liposome was successfully obtained in examples 1 to 13. The appearance, the average particle size and the glabridin content of the alundum plant-extracted aqueous glabridin liposome prepared in most of the examples are basically not different from those of the liposome prepared by the film method of comparative examples 2-3, which shows that the performance of the liposome prepared by the preparation method of the embodiment of the invention is basically consistent with that of the liposome prepared by the film method. Example 9 compared to the glabridin liposome prepared in comparative example 1, there was substantially no difference in appearance, average particle size, and glabridin content, which indicates that the effect of liposome prepared using the plant extract of artemisia annua instead of pure water in the example was substantially uniform, and the liposome was able to replace pure water as a solvent. The glabridin liposome prepared by the liposome preparation method of comparative example 4 is in a yellow turbid state in appearance, and insoluble particles exist; the sample has larger particle size, the encapsulation efficiency of the glabridin medicament is obviously reduced, which shows that when the temperature is insufficient, the phospholipid phase cannot be fully converted into the liquid crystal phase, and fat-soluble medicaments such as glabridin and the like are difficult to fully react with phospholipid molecules, so that the particle size is increased, the encapsulation efficiency is reduced, and the quality of a liposome preparation is reduced.

Table 2 characterization of glabridin liposomes for examples and comparative examples

Test example 2

The glabridin liposomes prepared in examples and comparative examples were subjected to preservative challenge test.

Inoculating certain amount of bacteria and fungi, and detecting the change of microbial quantity according to the detection method of the preservative efficacy test of the microorganisms in United states Pharmacopeia USP32<51> at intervals of 0 day, 7 days, 14 days, 21 days and 28 days, wherein the test results are shown in Table 3.

The national standard GB7918.2-87 cosmetic microbial standard test method is adopted to carry out microbial detection, and the test results are shown in Table 4.

As can be seen from tables 3 and 4, the glabridin liposome prepared in the examples had inhibitory effect on microorganisms and had certain antiseptic effect.

Table 3 example and comparative example glabridin liposomes preservative challenge test results

Table 4 examples and comparative examples microbial test results of glabridin liposomes

Test example 3

The glabridin liposome prepared in example 9 and comparative example 1 was tested for the moisturizing ability of the sample in an animal skin dryness model.

The experimental method comprises the following steps:

the skin of the neck and the back of the mouse is shaved and depilated, so that the skin of the neck and the back is fully exposed. Blank groups were not treated, model and experimental groups (including positive and sample groups) were run using a volume ratio of 1: 1, wiping the bare skin of the mouse by using a mixed solution of acetone and ether, and smearing corresponding samples (a blank group is smeared with a propylene glycol solution at 200 uL/piece/time, a model group is smeared with a propylene glycol solution at 200 uL/piece/time, a positive group is smeared with a propylene glycol solution of 0.5% hyaluronic acid at 200 uL/piece/time, and a sample group is smeared with the glabridin liposome prepared in the comparative example 1, the glabridin liposome prepared in the example 9 and the glabridin extract prepared in the example 1 at 200 uL/piece/time) for 7 days and 1 time per day. The mice were sacrificed and the skin tissue on the back of the neck was fixed with paraformaldehyde for 24h, sectioned with paraffin embedding, and the skin was stored in a refrigerator at-80 ℃. Measuring the protein content of AQP3 by an immunohistochemical method; the expression of Claudin1 gene was detected using fluorescent quantitative PCR. (AQP3 is transmembrane protein existing in human body, and participates in oxidative stress process of skin by regulating metabolism of human body water and lipid, the expression level and distribution of AQP3 can determine the water content of xerotic skin; Claudin1 protein is a rope constituting cells and is key for preventing dehydration inactivation, QRT-PCR is to adopt immunofluorescence method to quantify Claudin1 gene expression level, and whether the water content of cells is sufficient is measured)

The higher the protein content of AQP3, the more moisture content the skin. As shown in fig. 8, the blank control group had a normal content of AQP3 in the skin without dry molding treatment, and after dry molding, the protein content of the model group was significantly reduced, the skin was in a dehydrated dry state, and the dry state of the skin was improved after applying the moisture-retaining sample. Wherein, after the glabridin liposome and the sodium nervone extract water are compounded, the moisturizing capability of the glabridin liposome in the example 9 is better than that of the glabridin liposome compounded by pure water in the comparative example 1.

The higher the expression level of Claudin1 protein, the more sufficient the moisture content of the skin. As shown in fig. 9, the blank control group was the normal expression level of skin protein without dry molding treatment, and after dry molding, the expression level of protein in the model group was significantly reduced, and the skin was in a dehydrated and dry state, and the skin dry state was improved after applying the sample containing moisture retention. Wherein, after the glabridin liposome and the sodium nervone extract water are compounded, the moisturizing capability of the glabridin liposome is better than that of the glabridin liposome compounded by pure water in the proportion 1 and the sodium nervone extract water in the embodiment 1, which shows that the glabridin and the sodium nervone extract water can be compounded to synergistically enhance the moisturizing effect of the liposome.

The above-mentioned embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the same; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.

Claims

1. Application of herba Artemisiae Annuae extract water in preparing moisturizing liposome is provided.

2. The use of claim 1, wherein said blumea balsamifera plant extract water is obtained by distilling blumea balsamifera leaves under low temperature vacuum conditions.

3. Use according to claim 1, characterized in that it comprises the following steps:

4. Use according to claim 3, wherein the shear of step 2 is applied at a rate of 8000rpm or more for a period of 30min or more.

5. The use of claim 3, wherein the temperature of said heat incubation of step 2 is above the phase transition temperature of said phospholipid.

6. Use according to claim 3, characterized in that the said substances are, in mass percent,

2-15% of phospholipid;

0.01-5% of a medicament;

0-5% of cholesterol;

the balance of the folium Artemisiae Argyi plant extract water.

7. The use of claim 3, wherein step 2, prior to obtaining the liposome emulsion, further comprises: homogenizing under high pressure;

the high-pressure homogenization specifically comprises the following steps: each cycle was carried out at a pressure of 400bar, 600bar, 800bar, 1000bar for 2 or more times.

8. The use of claim 3, wherein step 1, before the mixing, further comprises: adding a surfactant and/or a polyol.

9. The use according to claim 8, wherein the surfactant is 0.1 to 20% by mass and the polyol is 5 to 60% by mass.

10. A liposome, wherein the liposome is prepared from a material comprising: medicine, phospholipid, cholesterol and water extracted from folium Artemisiae Argyi.