阅读说明：本技术 一种含甘草提取物的乳霜 (A cream containing Glycyrrhrizae radix extract ) 是由 鲍佳 金利伟 鲍益平 李扬 于 2021-08-02 设计创作，主要内容包括：本发明涉及一种提高甘草提取物稳定性、水溶性和透皮吸收性能的乳霜。一种含甘草提取物的乳霜,包括水、甘草提取物和辅料,所述辅料包括稳定剂和促渗剂。通过添加水溶性β-环糊精衍生物,提高乳霜中光果甘草提取物的稳定性；通过添加生物促渗剂短肽,在提高乳霜皮肤渗透性的同时进一步提高乳霜的稳定性。(The invention relates to a cream for improving the stability, water solubility and transdermal absorption performance of a liquorice extract. A cream containing Glycyrrhrizae radix extract comprises water, Glycyrrhrizae radix extract and adjuvants, wherein the adjuvants include stabilizer and penetration enhancer. The stability of the glycyrrhiza glabra extract in the cream is improved by adding water-soluble beta-cyclodextrin derivatives; by adding the short peptide of the biological penetration enhancer, the skin permeability of the cream is improved, and the stability of the cream is further improved.)

1. A cream containing licorice extract comprises water, licorice extract and auxiliary materials, and is characterized in that the auxiliary materials comprise a stabilizer and a penetration enhancer.

2. The licorice extract-containing cream according to claim 1, wherein the licorice extract is glycyrrhiza glabra extract.

3. The cream containing licorice extract according to claim 1, wherein the stabilizer is a water-soluble β -cyclodextrin derivative.

4. The cream containing licorice extract according to claim 3, wherein the water-soluble β -cyclodextrin derivative is one of hydroxypropyl- β -cyclodextrin and dialdehyde- β -cyclodextrin.

5. The cream containing licorice extract according to claim 3, wherein the water-soluble β -cyclodextrin derivative is a combination of hydroxypropyl- β -cyclodextrin and dialdehyde- β -cyclodextrin, and the weight ratio of hydroxypropyl- β -cyclodextrin to dialdehyde- β -cyclodextrin is 1-3: 1.

6. the cream containing licorice extract according to claim 1, wherein the penetration enhancer is a short peptide consisting of less than 30 amino acid residues.

7. The cream containing licorice extract according to claim 6, wherein the short peptide is one or a combination of Pep-1 peptide, T2 peptide and TAT peptide.

8. The licorice extract-containing cream according to claim 1, characterized in that the ratio by weight of licorice extract: a stabilizer: the penetration enhancer is 3: 2: 1.

9. the cream containing licorice extract according to claim 1, wherein the adjuvant further comprises a humectant, a keratolytic agent, an emollient, an emulsifier, an emulsion stabilizer, an antioxidant, a preservative, a chelating agent, and an aromatic.

10. A method for preparing a cream containing a licorice extract according to claim 1, characterized by sequentially adopting the following steps:

(1) preparing an aqueous phase pot, adding deionized water into the aqueous phase pot, adding glycyrrhiza glabra extract, hydroxypropyl-beta-cyclodextrin, dialdehyde-beta-cyclodextrin, T2 peptide, humectant, cutin softener and chelating agent into the aqueous phase pot, heating to 90-100 ℃ under stirring, maintaining for 10-30min for sterilization, and cooling to 70-80 ℃ for later use;

(2) preparing an oil phase pot, namely putting an emulsifier, an emulsion stabilizer, an antioxidant and an emollient into the oil phase pot, and heating to the appropriate temperature of 80-85 ℃ until the emulsifier, the emulsion stabilizer, the antioxidant and the emollient are completely dissolved;

(3) slowly adding the oil phase completed in the step (2) into the water phase completed in the step (1), then putting into an emulsifying pot preheated to 60-70 ℃, stirring to a proper emulsifying state, and then cooling to 30-55 ℃ for later use;

(4) adding aromatic and antiseptic, stirring, and aging to obtain cream.

Technical Field

The invention relates to a cream for improving the stability, water solubility and transdermal absorption performance of a liquorice extract.

Background

Age spots are common in the elderly population, and as the age of the population increases, the skin ages more rapidly, and brown or black patches and plaques develop, and the surface becomes rough and keratotic, and may occur anywhere in the body. Meanwhile, the skin water locking function of the middle-aged and the elderly is reduced, and the phenomenon of chronic physiological dehydration can occur, so that the skin is dry and wrinkles are increased. The skin of the middle-aged and the elderly has slow metabolism, poor moisture retention and poor flexibility, is not easy to adapt to seasonal changes, and is easy to generate dry cracks. The middle-aged and the elderly often lack proper skin care, and in addition, the blood circulation is poor due to insufficient sleep, fatigue and the like, the skin defense capability is reduced, and the skin problem is more likely to occur.

The glycyrrhiza glabra extract contains various active ingredients, mainly contains flavonoid substances such as glycyrrhizic acid and the like, can regulate the immune function of skin, enhance the disease resistance of the skin, eliminate inflammation, prevent allergy, clean the skin and simultaneously relieve the toxic and side effects of cosmetics and other external factors on the skin. In addition, it can effectively inhibit the activation of tyrosinase and prevent the production of melanin. For example, the patent applicant discloses a skin-care anti-cracking cream for middle-aged and elderly people (Chinese patent publication No. CN108158934A), which is mainly used for skin care and anti-cracking of skin.

The applicant finds that flavonoid substances such as glycyrrhizic acid and the like, which are main active ingredients contained in the glycyrrhiza glabra extract, are difficult to dissolve in water, and the glycyrrhizic acid is poor in stability, and is easy to decompose in light or acid-base environments, so that the glycyrrhiza glabra extract is not favorable for long-term use. For example, as disclosed in the chinese patent publication No. CN1520811A, the oil-soluble licorice extract obtained by using glycyrrhiza glabra as an extraction raw material contains glycyrrhizic acid as a main active ingredient, and a cream containing the glycyrrhizic acid is colored and smells after being stored for a long time.

Disclosure of Invention

The invention aims to solve the problems of poor stability and poor skin permeability of the cream containing the liquorice extract; the stability of the glycyrrhiza glabra extract in the cream is improved by adding water-soluble beta-cyclodextrin derivatives; by adding the short peptide of the biological penetration enhancer, the skin permeability of the cream is improved, and the stability of the cream is further improved.

In order to achieve the above object, the present invention adopts the following technical solutions:

a cream containing Glycyrrhrizae radix extract comprises water, Glycyrrhrizae radix extract and adjuvants, wherein the adjuvants include stabilizer and penetration enhancer.

Preferably, the licorice extract is a glycyrrhiza glabra extract.

Preferably, the stabilizer is a water-soluble β -cyclodextrin derivative.

Preferably, the water-soluble beta-cyclodextrin derivative is one of hydroxypropyl-beta-cyclodextrin and dialdehyde-beta-cyclodextrin.

Preferably, the water-soluble beta-cyclodextrin derivative is a composition of hydroxypropyl-beta-cyclodextrin and dialdehyde-beta-cyclodextrin, and the weight ratio of the hydroxypropyl-beta-cyclodextrin to the dialdehyde-beta-cyclodextrin is 1-3: 1.

preferably, the penetration enhancer is a short peptide consisting of less than 30 amino acid residues. Further, the short peptide is one or a combination of Pep-1 peptide, T2 peptide and TAT peptide.

Preferably, the weight ratio of the liquorice extract: a stabilizer: the penetration enhancer is 3: 2: 1.

preferably, the auxiliary materials also comprise a humectant, a cutin softener, an emollient, an emulsifier, an emulsion stabilizer, an antioxidant, a preservative, a chelating agent and an aromatic.

The application also discloses a preparation method of the cream containing the liquorice extract, which sequentially comprises the following steps:

(1) preparing an aqueous phase pot, adding deionized water into the aqueous phase pot, adding glycyrrhiza glabra extract, hydroxypropyl-beta-cyclodextrin, dialdehyde-beta-cyclodextrin, T2 peptide, humectant, cutin softener and chelating agent into the aqueous phase pot, heating to 90-100 ℃ under stirring, maintaining for 10-30min for sterilization, and cooling to 70-80 ℃ for later use;

(2) preparing an oil phase pot, namely putting an emulsifier, an emulsion stabilizer, an antioxidant, an emollient and an antioxidant into the oil phase pot, and heating to the proper temperature of 80-85 ℃ until the emulsifier, the emulsion stabilizer, the antioxidant, the emollient and the antioxidant are completely dissolved;

(3) slowly adding the oil phase completed in the step (2) into the water phase completed in the step (1), then putting into an emulsifying pot preheated to 60-70 ℃, stirring to a proper emulsifying state, and then cooling to 30-55 ℃ for later use;

(4) adding aromatic and antiseptic, stirring, and aging to obtain cream.

The invention has the technical effects that the stability of the glycyrrhiza glabra extract in cream is improved by adding water-soluble beta-cyclodextrin derivatives, particularly a composition of hydroxypropyl-beta-cyclodextrin and dialdehyde-beta-cyclodextrin, and glycyrrhizic acid to form an inclusion compound; and the stability of the cream is further improved while the skin permeability of the cream is improved by adding a penetration enhancer, particularly T2 peptide. In conclusion, the cream containing the licorice extract has good stability and skin permeability.

Drawings

FIG. 1: standard curve diagram of glycyrrhizic acid solution.

FIG. 2: and (3) adding different stabilizers to test the change condition of the content of the glycyrrhizic acid in the sample.

FIG. 3: and (3) adding test samples of different penetration promoters into the in vitro transdermal diffusion experiment for 24h to obtain the total penetration result.

FIG. 4: and (3) adding different penetration enhancers to test the change condition of the content of the glycyrrhizic acid in the sample.

FIG. 5: the transdermal permeation rate of the finished products with different proportions in an in-vitro transdermal diffusion experiment.

Detailed Description

The technical solution of the present invention is further described below by means of specific examples.

The main experimental materials: and (3) liquorice extract: chengdu Gelipu Biotech limited; glycyrrhizic acid control: tianjin Tech technologies, Inc.; penetration enhancing peptides Pep-1, T2 and TAT: doudu Yunshi chemical Co., Ltd; beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin, dialdehyde-beta-cyclodextrin and other auxiliary materials are all commercial products.

Example 1

The formula of the cream containing the licorice extract comprises, by weight, 75.86g of deionized water, 9g of the glycyrrhiza glabra extract, 4g of hydroxypropyl-beta-cyclodextrin, 2g of dialdehyde-beta-cyclodextrin, 3g of T2 peptide, 12g of a humectant, 15g of a cutin softener, 0.02g of a chelating agent, 1.5g of an emulsifier, 2g of an emulsion stabilizer, 4g of an emollient, 1.5g of an antioxidant, 0.1g of an essence and 0.02g of a preservative.

Wherein, the auxiliary materials are specifically selected as follows:

humectant: water-soluble glycerin; keratolytic agent: urea; chelating agent: disodium EDTA; emulsifier: glyceryl stearate and stearyl polyoxyethylene 21 ether; emulsion stabilizer: stearic acid; and (3) an emollient: petrolatum, dimethicone and olive oil; antioxidant: tocopheryl acetate; aromatic agent: an essence; preservative: phenoxyethanol.

The preparation method of the finished cream product containing the liquorice extract adopts the raw materials and sequentially comprises the following steps: (1) preparing an aqueous phase pot, namely adding deionized water into the aqueous phase pot, adding the glycyrrhiza glabra extract, hydroxypropyl-beta-cyclodextrin, dialdehyde-beta-cyclodextrin, T2 peptide, humectant, cutin softener and chelating agent into the aqueous phase pot, heating to 90-100 ℃ under stirring, maintaining for 10-30min for sterilization, and cooling to 70-80 ℃ for later use; (2) preparing an oil phase pot, namely putting an emulsifier, an emulsion stabilizer, an antioxidant, an emollient and an antioxidant into the oil phase pot, and heating to the proper temperature of 80-85 ℃ until the emulsifier, the emulsion stabilizer, the antioxidant, the emollient and the antioxidant are completely dissolved; (3) slowly adding the oil phase completed in the step (2) into the water phase completed in the step (1), then putting into an emulsifying pot preheated to 60-70 ℃, stirring to a proper emulsifying state, and then cooling to 30-55 ℃ for later use; (4) adding aromatic and antiseptic, stirring, and aging to obtain cream. And canning and packaging.

Comparative example 1 stability test

Test samples were prepared according to the formulation of table 1, with the raw materials and preparation process essentially the same as in example 1, except that no penetration enhancer was added to the adjuvants. In the experiment, a test sample is placed in an environment with 48 +/-2 ℃ and strong light irradiation, the sample is kept and the change of one-time characters is observed at 0, 5, 10, 15, 20, 25 and 30 days, and the content of the glycyrrhizic acid in the cream is monitored.

Table 1 comparative example 1 test sample formulation table

Note: "√" indicates the inclusion of a certain component, and "X" indicates the exclusion of a certain component.

1.1 analytical methods: HPLC method

Chromatographic conditions are as follows: diamond^TM C₁₈Chromatography columns (250mm x 4.6mm i.d.,5 μm); the mobile phase is acetonitrile and 1% glacial acetic acid water solution (55: 45), the detection wavelength is 280nm, the sample injection amount is 30 mu L, and the flow rate is 1 mL/min; the column temperature was 30 ℃.

Preparation of a standard solution: taking a proper amount of optical glycyrrhizic acid standard substance, precisely weighing, dissolving with mobile phase, fixing volume to 100.0 μ g/ml standard substance solution, and storing at-20 deg.C as standard stock solution.

Establishing a standard curve: preparing standard substance glycyrrhizic acid into 0.01, 0.1, 0.5, 1.0, 2.0, 5.0, 10.0, 20.0, 40.0 μ g/ml series standard solution, injecting sample according to the above chromatographic conditions, drawing standard curve with peak area to concentration, linear range of 0.01-40.0 μ g/ml, standard curve equation of A being 33067m +1211.1, R²0.9996 (where m is the concentration of glycyrrhizic acid and a is the peak area) as shown in figure 1.

The appearance stability of sample No. 1 appeared yellowish and slightly smelled on day 5, and a small amount of bubbles were also generated; sample No. 2 had a slight yellow color starting on day 20; sample No. 3 and sample No. 4 developed a coloration phenomenon on day 25; by day 30, the appearance of sample No. 5 was still substantially unchanged. As shown in fig. 2, the content of glycyrrhizic acid in sample No. 5, to which hydroxypropyl- β -cyclodextrin and dialdehyde- β -cyclodextrin were added, was substantially unchanged, and stability was the best. The content of glycyrrhizic acid in sample No. 3 and sample No. 4 is slightly reduced, while the content of glycyrrhizic acid in sample No. 2 added with beta-cyclodextrin is reduced at a faster rate than in sample No. 3 and sample No. 4, but still remains in the range where activity can be exerted. The content of glycyrrhizic acid in sample No. 1 is reduced most obviously.

In addition, a sample No. 6 and a sample No. 7 are also prepared, the formula components and the preparation process of the sample No. 6 and the sample No. 7 are the same as those of the sample No. 5, and the difference is only that the dosage of hydroxypropyl-beta-cyclodextrin and dialdehyde-beta-cyclodextrin is different, wherein the dosage of hydroxypropyl-beta-cyclodextrin and the dosage of dialdehyde-beta-cyclodextrin in the sample No. 6 are both 3 g; the sample No. 7 contained 4.5g of hydroxypropyl-. beta. -cyclodextrin and 1.5g of dialdehyde-. beta. -cyclodextrin. The observation, detection and analysis were carried out in the same experimental manner as described above. The appearance stability of the sample No. 6 and the sample No. 7 is not obviously different from that of the sample No. 5, and the yellowing and the odor are not generated in 30 days. According to HPLC detection, compared with the sample No. 0, the content of the glycyrrhizic acid in the sample No. 5 on the day 30 is reduced by only 0.25%, the content of the glycyrrhizic acid in the sample No. 6 is reduced by 8.58%, the content of the glycyrrhizic acid in the sample No. 7 is reduced by 9.51%, and both the reduction ranges are larger than those of the sample No. 5. This experiment thus demonstrates that when hydroxypropyl- β -cyclodextrin and dialdehyde- β -cyclodextrin are added in a ratio of 2: the sample stability at 1 is best.

Comparative example 2 in vitro transdermal diffusion experiment

Sample No. 8 and sample No. 9 were prepared according to the formulation and preparation method of example 1, and sample No. 8 and sample No. 9 were different from example 1 only in the penetration enhancer.

Table 2 comparative example 2 test sample formulation table

Composition of matter	Sample 8	Sample 9	EXAMPLE 1 finished product
				Glycyrrhiza glabra extract	√	√	√
Pep-1 peptides	√	×	×
				T2 peptide	×	×	√
TAT peptides	×	√	×
				Other auxiliary materials	√	√	√

Note: "√" indicates the inclusion of a certain component, and "X" indicates the exclusion of a certain component.

The analytical method was the same as the HPLC method described in comparative example 1.

The Content (CX) in the permeated solution was calculated from the peak area of the photo-glycyrrhizic acid in units of μ g/ml.

The experimental procedure was as follows: the pre-treated mouse belly depilated skin was mounted between the dosing and receiving wells of the Frangz diffusion cell and 1g of all samples and controls prepared in comparative example 2 were applied evenly to the skin of the mouse. The receiving solution is 20% ethanol physiological saline solution, and the dermis side is connected with the receiving solutionThe receiving liquid is completely contacted, and care is taken to remove bubbles on the contact surface. The volume of the receiving solution is 10ml, and the effective diffusion area is 4.12cm²The test temperature was 37. + -. 1 ℃ and the stirring speed was 800 rpm. Sampling 1ml at 2, 4, 6, 8 and 24h (simultaneously supplementing equal amount of receiving solution at the same temperature), filtering, and determining content of glycyrrhizic acid by the above analysis method. Each sample was tested in parallel 3 times, and the cumulative permeation amount per unit area Q was calculated according to the following formula (1)₁/ug·cm^-2And the formula (2) is used for calculating the percutaneous steady state permeation rate Js/ug cm^-2/h^-1

Formula (1): q₁(content (CX) × 10.0)/4.12

Formula (2): js ═ Q₁T and t are respectively 2, 4, 6, 8 and 24 h.

As shown in FIG. 3, the total 24-hour transmittance of sample No. 8 was 34.50ug cm^-2The total 4-hour permeability of the finished product of example 1 was 35.04ug cm^-2Sample No. 9 had a total 24-hour permeability of 35.12 ug/cm^-2. The experimental result proves that the skin permeability of the cream can be improved by respectively adding three different short peptide penetration enhancers, but the effects of improving the transdermal absorbability of the cream are not good.

The stability tests were performed on sample No. 8, sample No. 9 and the finished product of example 1 in this comparative example, and the test methods were the same as those described above. As a result, as shown in fig. 4, the content of glycyrrhizic acid in sample No. 8 and sample No. 9 decreased greatly, but the content of glycyrrhizic acid in the finished product of example 1 was almost constant and better than that in sample No. 5 in comparative example 1. The experiment proves that the T2 peptide added into the cream containing the liquorice extract can not only improve the permeability of the cream, but also improve the stability of the cream.

Comparative example 3

Preparing a finished product No. 2 and a finished product No. 3 according to the formula and the preparation method of the example 1, wherein the finished product No. 2 is different from the finished product No. 1 only in the dosage of the stabilizer and the penetration enhancer, wherein the dosage of hydroxypropyl-beta-cyclodextrin in the finished product No. 2 is 3.5g, the dosage of dialdehyde-beta-cyclodextrin in the finished product No. 1 is 1.5g, and the dosage of T2 peptide in the finished product No. 4 g; the finished product No. 3 contains 4.5g of hydroxypropyl-beta-cyclodextrin, 2.5g of dialdehyde-beta-cyclodextrin and 2g of T2 peptide.

Stability tests were performed on the finished product of example 1, the finished product of No. 2 and the finished product of No. 3 by HPLC. On day 15, the content of glycyrrhizic acid in finished product No. 2 was reduced by 0.12%, compared with that on day 0, that in finished product No. 3 was reduced by 0.13%, and that in finished product example 1 was reduced by 0.015%. On day 30, the content of glycyrrhizic acid in the finished product No. 2 is reduced by 0.38 percent compared with that on day 0, the content of glycyrrhizic acid in the finished product No. 3 is reduced by 0.35 percent, the content of glycyrrhizic acid in the finished product in the example 1 is reduced by only 0.03 percent, and the stability is better than that of the finished products No. 2 and No. 3. Thus, this experiment demonstrates that glycyrrhiza glabra extract: a stabilizer: the penetration enhancer is 3: 2: cream stability was best at 1.

The finished products of example 1, No. 2 and No. 3 were further subjected to an in vitro transdermal diffusion test, with t vs. Q₁Plotted, as shown in fig. 5, the steady state release rate was highest for the finished product of example 1, superior to that of finished product No. 2 and finished product No. 3. In summary, it is further demonstrated that in the finished product simultaneously added with hydroxypropyl- β -cyclodextrin, dialdehyde- β -cyclodextrin composition and T2 peptide, glycyrrhiza glabra extract is obtained: a stabilizer: the penetration enhancer is 3: 2: the transdermal quantity is the largest at 1, and the bioavailability is better.

Because the main active ingredients of the different licorice raw material extracts are different, the glycyrrhiza glabra selected by the invention contains the main active ingredient of glycyrrhizic acid which has the effects of inhibiting tyrosinase activity, removing oxygen free radicals and resisting inflammation, and can be applied to cream for skin care. The photo-glycyrrhizic acid can be decomposed with time, so that the activity of the cream containing the component is reduced until the cream is not effective. The beta-cyclodextrin has a special cyclic structure, can stabilize substances which are easy to volatilize, oxidize, photolyze and hydrolyze, can change the chemical properties of the substances, and can solvate and emulsify the substances which are difficult to dissolve in water. Beta-cyclodextrin derivatives are more widely used because of their improved toxicity, physicochemical properties and solubility. Wherein the beta-cyclodextrin derivative is mainly divided into three types of water solubility, hydrophobicity and ionic type, and the water-soluble beta-cyclodextrin derivative has better solubilization effect on substances with strong lipophilicity. In order to improve the stability of the glycyrrhizic acid and prolong the service life of the cream, the invention adds the water-soluble beta-cyclodextrin derivative, in particular to the hydroxypropyl-beta-cyclodextrin and dialdehyde-beta-cyclodextrin composition to form an inclusion compound with the glycyrrhizic acid, and adopts reasonable proportion to improve the stability of the glycyrrhizic acid. Because the cream containing the licorice extract has poor permeability when being externally applied to skin, the invention adds the biological penetration enhancer to promote the skin permeability and improve the bioavailability, wherein the selected Pep-1 peptide, T2 peptide and TAT peptide have good penetration enhancing effect and little difference on the product, and tests show that the stability of the cream can be enhanced while the penetration enhancing effect is ensured only by adding the T2 peptide. Finally, the stabilizer in the invention is selected from a composition of hydroxypropyl-beta-cyclodextrin and dialdehyde-beta-cyclodextrin, the penetration enhancer is selected from T2 peptide, and when the glycyrrhizic acid in the cream: a stabilizer: the weight ratio of the penetration enhancer is 3: 2: 1, optimal stability and permeability are achieved.