Glycyrrhizic acid and bacterial cellulose compound for promoting growth and development of skin hair follicles

Abstract

The invention relates to a glycyrrhizic acid and bacterial cellulose compound for promoting the growth and development of skin hair follicles, in particular to a glycyrrhizic acid and bacterial cellulose compound for promoting the growth and development of the skin hair follicles, which comprises glycyrrhizic acid and bacterial cellulose.

Description

Technical Field

[0001] The present invention relates to a bacterial cellulose composite, and particularly to a glycyrrhizic acid bacterial cellulose composite and a preparation method thereof. Background Art

[0002] Bacterial cellulose is a relatively novel nanomaterial that has been less applied to drug delivery. It has been proven to have a variety of special physical and chemical properties, including biodegradability, non-toxicity, high elastic modulus, high specific surface area, low density, non-abrasiveness, easy surface functionalization, high purity and crystallinity in chemical composition, high degree of polymerization (2000 - 8000), and high hardness. At the same time, bacterial cellulose is an inert material, so there is less direct research on bacterial cellulose as a drug carrier. Although glycyrrhizic acid is a commonly used drug in clinical practice with anti-inflammatory and detoxifying effects, glycyrrhizic acid has no practical application background for topical drug use.

[0003] Therefore, the clinical treatment of hair loss remains a difficult problem that the industry urgently needs to overcome. Summary of the Invention

[0004] In view of the various defects of the above-mentioned prior art, the present invention provides a glycyrrhizic acid bacterial cellulose composite that can promote the growth and development of hair follicles and a preparation method thereof. The glycyrrhizic acid bacterial cellulose composite includes glycyrrhizic acid and bacterial cellulose. When the glycyrrhizic acid bacterial cellulose composite is topically applied to the skin, it can promote the growth and development of hair follicles in the skin, which is beneficial to the clinical treatment of hair loss and the promotion of hair growth.

[0005] In a specific embodiment, the bacterial cellulose is derived from Acetobacterium Balch and has a molecular weight of 50,000 to 2,500,000.

[0006] In a specific embodiment, the bacterial cellulose has 300 to 15,000 glucose units.

[0007] In a specific embodiment, the mass ratio of glycyrrhizic acid to bacterial cellulose is 1:0.1 to 10; preferably, the mass ratio of glycyrrhizic acid to bacterial cellulose is 1:5.

[0008] The present invention also provides a method for preparing the aforementioned glycyrrhizic acid bacterial cellulose composite, including the following steps: 1) mixing water and freeze-dried bacterial cellulose to obtain hydrated bacterial cellulose; 2) dissolving glycyrrhizic acid in water to obtain a glycyrrhizic acid aqueous solution; and 3) dropping the glycyrrhizic acid aqueous solution into the hydrated bacterial cellulose to obtain the glycyrrhizic acid bacterial cellulose composite.

[0009] In a specific embodiment, the hydrated bacterial cellulose and the glycyrrhizic acid aqueous solution are each stirred into a uniform solution.

[0010] In a specific embodiment, the stirred aqueous bacterial cellulose and glycyrrhizic acid aqueous solution are each further subjected to ultrasonic treatment.

[0011] In a specific embodiment, the dropping is carried out under ultrasonic waves.

[0012] In a specific embodiment, the freeze-dried bacterial cellulose and water are mixed at a weight ratio of 1:10 to 15; preferably, the glycyrrhizic acid and water are mixed at a weight ratio of 1:0.5 to 1.5.

[0013] In a specific embodiment, the mass ratio of the glycyrrhizic acid to the bacterial cellulose in the glycyrrhizic acid-bacterial cellulose complex is 1:0.1 to 10; preferably, the mass ratio of the glycyrrhizic acid to the bacterial cellulose in the glycyrrhizic acid-bacterial cellulose complex is 1:5.

[0014] Specifically, the glycyrrhizic acid-bacterial cellulose complex provided by the present invention is white and viscous, and the bacterial cellulose structure is loaded with glycyrrhizic acid. The present invention utilizes the amphiphilicity of glycyrrhizic acid and the biocompatibility, non-toxicity and reticular structure of bacterial cellulose to obtain a substance that can significantly promote the growth and development of hair follicles in the skin. According to the research results of the promoting effect on the growth and development of hair follicles in the skin and combined with the requirements of pharmaceutical preparations, the most suitable amounts of glycyrrhizic acid and bacterial cellulose can be further selected and the feeding ratio with the best promoting effect on the growth and development of skin hair follicles can be used to prepare related topical pharmaceutical preparations. Since glycyrrhizic acid itself is an amphiphilic compound and the characteristics of bacterial cellulose itself include non-toxicity, high biocompatibility and the use background as a carrier for skin care products, the present invention can be applied to the development and research of transdermal topical pharmaceutical preparations for treating alopecia and promoting hair growth. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The embodiments of the present application are illustrated by exemplary reference drawings:

[0016] Figures 1A to 1C SEM diagrams of Comparative Example 1, Comparative Example 3 and Example 5 are respectively shown.

[0017] Figure 2 The glycyrrhizic acid contents of Comparative Example 2, Example 1, Example 5 and Example 10 at different time periods are shown.

[0018] Figure 3 The glycyrrhizic acid contents of Comparative Example 2, Example 1, Example 3 and Example 5 at the skin penetration end point are shown.

[0019] Figure 4A and Figure 4BThese are histological staining diagrams of hair follicles in the skin untreated and treated with the glycyrrhizic acid bacterial cellulose complex of Example 10. The skin before penetration (left figure) and at the end point of penetration (right figure) was stained with toluidine blue and hematoxylin-eosin (HE), respectively. Detailed implementation mode

[0020] The following specific embodiments illustrate the implementation modes of the present application. Those skilled in the art can easily understand the advantages and effects of the present application from the content described in this specification. The present application can also be implemented or applied through other different implementation modes. All details in this specification can also be given different modifications and changes based on different viewpoints and applications without departing from the spirit described in the present application. In addition, all ranges and values herein are inclusive and combinable. Any numerical value or point within the ranges described herein, such as any integer, can be used as the minimum or maximum value to derive sub-ranges, etc.

[0021] Glycyrrhizic acid is a commonly used clinical drug with functions such as anti-inflammatory and detoxifying effects. It belongs to triterpenoid compounds and has amphiphilicity, thus showing the characteristics of a surfactant. Aggregates or micelles of glycyrrhizic acid can form host-guest inclusion complexes with hydrophobic drugs, which can effectively increase the solubility of drugs and avoid the precipitation of drugs.

[0022] Bacterial cellulose has characteristics such as biodegradability, non-toxicity, high-elastic modulus, high specific surface area, low density, non-abrasiveness, easy surface functionalization, high purity and crystallinity in chemical composition, high degree of polymerization (2000 - 8000), and high hardness. However, bacterial cellulose is extremely inert, and there are few related studies on applying bacterial cellulose to local drug carriers including promoting hair growth.

[0023] Therefore, the present invention utilizes the amphiphilicity of glycyrrhizic acid to develop bacterial cellulose as a local drug carrier. Through research, it is found that the glycyrrhizic acid bacterial cellulose complex can promote the growth and development of hair follicles in the skin. As described above, the present invention uses glycyrrhizic acid as the drug to be delivered and bacterial cellulose as the carrier. Due to the amphiphilicity of glycyrrhizic acid, glycyrrhizic acid is loaded into the network structure of bacterial cellulose to generate a new structure, and the formed glycyrrhizic acid bacterial cellulose complex can significantly promote the growth and development of hair follicles in the skin.

[0024] In a specific embodiment, the glycyrrhizic acid is loaded on the bacterial cellulose to form a glycyrrhizic acid bacterial cellulose complex.

[0025] In a specific embodiment, the bacterial cellulose is derived from Acetobacterium Balch, which has a molecular weight of 50,000 to 2,500,000, a molecular weight of 100,000 to 2,500,000, a molecular weight of 500,000 to 2,500,000, a molecular weight of 1,000,000 to 2,500,000, a molecular weight of 1,500,000 to 2,500,000, a molecular weight of 2,000,000 to 2,500,000, a molecular weight of 50,000 to 2,000,000, a molecular weight of 50,000 to 1,500,000, a molecular weight of 50,000 to 1,000,000, a molecular weight of 50,000 to 500,000 or a molecular weight of 50,000 to 100,000. For example, a molecular weight of 50,000, 100,000, 150,000, 200,000, 250,000, 300,000, 350,000, 400,000, 450,000, 500,000, 550,000, 600,000, 650,000, 700,000, 750,000, 800,000, 850,000, 900,000, 950,000, 1,000,000, 1,500,000, 2,000,000 or 2,500,000. In another specific embodiment, the bacterial cellulose is obtained by fermentation of Acetobacterium.

[0026] In a specific embodiment, the bacterial cellulose has 300 to 15,000 glucosyl groups, 1,000 to 15,000 glucosyl groups, 2,000 to 15,000 glucosyl groups, 3,000 to 15,000 glucosyl groups, 4,000 to 15,000 glucosyl groups, 5,000 to 15,000 glucosyl groups, 6,000 to 15,000 glucosyl groups, 7,000 to 15,000 glucosyl groups, 8,000 to 15,000 glucosyl groups, 9,000 to 15,000 glucosyl groups, 10,000 to 15,000 glucosyl groups, 11,000 to 15,000 glucosyl groups, 12,000 to 15,000 glucosyl groups, 13,000 to 15,000 glucosyl groups, 14,000 to 15,000 glucosyl groups, 300 to 14,000 glucosyl groups, 300 to 13,000 glucosyl groups, 300 to 12,000 glucosyl groups, 300 to 11,000 glucosyl groups, 300 to 10,000 glucosyl groups, 300 to 9,000 glucosyl groups, 300 to 8,000 glucosyl groups, 300 to 7,000 glucosyl groups, 300 to 6,000 glucosyl groups, 300 to 5,000 glucosyl groups, 300 to 4,000 glucosyl groups, 300 to 3,000 glucosyl groups, 300 to 2,000 glucosyl groups or 300 to 1,000 glucosyl groups. For example, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1,000, 1,500, 2,000, 2,500, 3,000, 3,500, 4,000, 4,500, 5,000, 5,500, 6,000, 6,500, 7,000, 7,500, 8,000, 8,500, 9,000, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500 or 15,000 glucosyl groups. Specifically, the bacterial cellulose has the chemical general formula of (C 6 H 10 O 5 ) n and is a polysaccharide composed of linear chains (glycosidic bonds) of hundreds to thousands of β(1→4)-linked D-glucose units. In other words, the bacterial cellulose is a macromolecular polysaccharide composed of D-glucose with β-1,4-glycosidic bonds.

[0027] In a specific embodiment, the mass ratio of glycyrrhizic acid to bacterial cellulose is from 1:0.1 to 10, for example, 1:0.1, 1:0.11, 1:0.13, 1:0.14, 1:0.17, 1:0.2, 1:0.3, 1:0.33, 1:0.4, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10. Preferably, the mass ratio of glycyrrhizic acid to bacterial cellulose is 1:5.

[0028] The method for preparing the glycyrrhizic acid - bacterial cellulose complex provided by the present invention comprises the following steps: 1) Mix water and freeze - dried bacterial cellulose to obtain hydrated bacterial cellulose; 2) Dissolve glycyrrhizic acid in water to obtain an aqueous glycyrrhizic acid solution; and 3) Dropwise add the aqueous glycyrrhizic acid solution into the hydrated bacterial cellulose to obtain the glycyrrhizic acid - bacterial cellulose complex.

[0029] In a specific embodiment, the freeze - dried bacterial cellulose and water are mixed at a weight ratio of 1:10 to 15, for example, a weight ratio of 1:10, 1:11, 1:12, 1:13, 1:14 or 1:15.

[0030] In a specific embodiment, the glycyrrhizic acid and water are mixed at a weight ratio of 1:0.5 to 1.5, for example, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1.0, 1:1.1, 1:1.2, 1:1.3, 1:1.4 or 1:1.5.

[0031] The following further elaborates on the present application through specific examples, but the scope of the present application is not limited by the examples.

[0032] Preparation Example: Glycyrrhizic Acid - Bacterial Cellulose Complex

[0033] Glycyrrhizic acid (93%, G810520 - 25g, Shanghai Macklin Biochemical Co., Ltd.) was added to distilled water, stirred evenly and then sonicated for 10 minutes to obtain an aqueous glycyrrhizic acid solution. At the same time, bacterial cellulose derived from Acetobacter (nano - bacterial cellulose frozen ingot, Jiaopeng Biotechnology Co., Ltd.) was added to distilled water, stirred evenly and then sonicated for 10 minutes to obtain hydrated bacterial cellulose; under ultrasonic conditions, the aqueous glycyrrhizic acid solution was dropped into the hydrated bacterial cellulose at a rate of 5 drops per 10 seconds, stirred evenly and then sonicated for 15 minutes to obtain the glycyrrhizic acid - bacterial cellulose complex.

[0034] According to the method of the foregoing preparation example, the bacterial cellulose complexes of Examples 1 to 19 were respectively prepared with the compositions shown in Table 1 below, and the mass ratios of glycyrrhizic acid and bacterial cellulose in the prepared glycyrrhizic acid - bacterial cellulose complexes (finished products) were listed.

[0035] Table 1: Examples 1 to 19

[0036]

[0037] Comparative Example 1: Glycyrrhizic acid aqueous solution

[0038] 10 mg of glycyrrhizic acid (93%, G810520 - 25 g, Shanghai Macklin Biochemical Co., Ltd.) was added to 1.6 mL of distilled water. After stirring evenly, it was sonicated for 10 minutes to obtain a glycyrrhizic acid aqueous solution.

[0039] Comparative Example 2: Hydrous bacterial cellulose

[0040] 50 mg of bacterial cellulose derived from Acetobacter (nano bacterial cellulose freeze-dried tablets, Jiaopeng Biotechnology Co., Ltd.) was added to 1.3 mL of distilled water. After stirring evenly, it was sonicated for 10 minutes to obtain hydrous bacterial cellulose.

[0041] Comparative Example 3: Baicalin bacterial cellulose

[0042] 10 mg of baicalin (8802695 - 5 g, Shanghai Macklin Biochemical Co., Ltd.) was added to 0.8 mL of distilled water. After stirring evenly, it was sonicated for 10 minutes to obtain a baicalin aqueous solution. At the same time, 100 mg of bacterial cellulose derived from Acetobacter (nano bacterial cellulose freeze-dried tablets, Jiaopeng Biotechnology Co., Ltd.) was added to 1.3 mL of distilled water. After stirring evenly, it was sonicated for 10 minutes to obtain hydrous bacterial cellulose; under ultrasonic conditions, the baicalin aqueous solution was added dropwise to the hydrous bacterial cellulose, stirred evenly and sonicated for 15 minutes, and then obtained a baicalin bacterial cellulose complex after rotary evaporation.

[0043] Please refer to Figures 1A to 1C ... Comparative Example 1, Comparative Example 3 and Example 5 were analyzed by a scanning electron microscope (FEI Quanta 400 FEI, America FEI scanning electron microscope), and the obtained SEM images were respectively shown in Figures 1A to 1C ... As shown in Figures 1A to 1C ... only the glycyrrhizic acid bacterial cellulose complex prepared in Example 5 has a structure in which glycyrrhizic acid is encapsulated in the reticular structure of bacterial cellulose.

[0044] To demonstrate the ability of different samples (i.e., Comparative Examples 1 to 3 and Examples 1 to 19) to penetrate the skin with glycyrrhizic acid, a skin penetration experiment was conducted using the dorsal skin of SD rats in a Franz cell diffusion cell. First, before the experiment, the skin (3 cm x 3 cm) in the diffusion cell was cleaned with 50% ethanol. Then, 200 mg of the sample was evenly applied to the cleaned skin, and after the experiment, the skin was cleaned with 20% ethanol to remove the surface glycyrrhizic acid. The absorption solutions were collected at 2, 4, and 6 hours respectively, and 0.1 mL of the absorption solution was mixed with 0.9 mL of methanol. Next, the mixed solution of the absorption solution and methanol was subjected to high performance liquid chromatography to determine the concentration of glycyrrhizic acid in the skin at different time periods.

[0045] Using Comparative Example 2, Example 1, Example 5, and Example 10 as samples respectively, the aforementioned skin penetration experiment and high performance liquid chromatography were carried out. Each sample was measured three times repetitively and the average value was taken. The results are shown in Figure 2 In addition, using Comparative Example 2, Example 1, Example 3, and Example 5 as samples respectively, the aforementioned skin penetration experiment and high performance liquid chromatography were carried out to determine the content of glycyrrhizic acid at the skin penetration end point (i.e., 360 minutes after the start of the experiment). From the results shown in Figure 2 and 3 it can be seen that the presence of glycyrrhizic acid was not detected in the absorption solution of Comparative Example 2; as the proportion of bacterial cellulose in the glycyrrhizic acid - bacterial cellulose complex increased, the cumulative penetration amount of glycyrrhizic acid increased, indicating that glycyrrhizic acid in the glycyrrhizic acid - bacterial cellulose complex with a higher proportion of bacterial cellulose had enhanced ability to penetrate the skin.

[0046] Please refer to Figure 4A and Figure 4B . Before the aforementioned skin penetration experiment and at the skin penetration end point, the skin was stained with toluidine blue (left figure) and HE staining (right figure) respectively, and the skin tissues without treatment and treated with the glycyrrhizic acid - bacterial cellulose complex of Example 10 were compared to observe the changes in the number and morphology of hair follicles in the skin of equal area. As shown in Figure 4A and Figure 4B , the glycyrrhizic acid - bacterial cellulose complex provided by the present invention can promote the growth and development of hair follicles in the skin.

[0047] In summary, the glycyrrhizic acid - bacterial cellulose complex of the present invention has glycyrrhizic acid in the three - dimensional network structure of bacterial cellulose, and can effectively promote the growth and development of hair follicles in the skin. In addition, the manufacturing process of the glycyrrhizic acid - bacterial cellulose complex provided by the present invention is simple, and the proportion of glycyrrhizic acid and bacterial cellulose contained in the complex can be easily adjusted, which actually has application prospects.

[0048] The above embodiments are only illustrative and not intended to limit the present application. Any person skilled in the art can modify and change the above embodiments without departing from the spirit and scope of the present application. Therefore, the scope of the claims of the present application is defined by the appended claims, and should be covered by the disclosed technical content as long as it does not affect the effect and implementation purpose of the present application.

Claims

1. A glycyrrhizic acid-bacterial cellulose complex for promoting the growth and development of skin hair follicles, comprising glycyrrhizic acid and bacterial cellulose.

2. The glycyrrhizic acid bacterial cellulose composite according to claim 1, characterized in that: The bacterial cellulose is derived from Acetobacterium Balch and has a molecular weight of 50,000 to 2,500,000.

3. The glycyrrhizic acid bacterial cellulose composite according to claim 1, characterized in that The bacterial cellulose has 300 to 15,000 glucose groups.

4. The glycyrrhizic acid bacterial cellulose composite according to any one of claims 1 to 3, characterized in that: The mass ratio of glycyrrhizic acid to bacterial cellulose is 1:0.1 to 10.

5. The glycyrrhizic acid bacterial cellulose composite according to any one of claims 1 to 3, characterized in that: The mass ratio of the glycyrrhizic acid to the bacterial cellulose is 1:

5.

6. A method for preparing the glycyrrhizic acid bacterial cellulose composite according to any one of claims 1 to 5, comprising the following steps: 1) mixing water and freeze-dried bacterial cellulose to obtain aqueous bacterial cellulose; 2) dissolving glycyrrhizic acid in water to obtain a glycyrrhizic acid aqueous solution; as well as 3) adding the glycyrrhizic acid aqueous solution dropwise to the aqueous bacterial cellulose to obtain the glycyrrhizic acid-bacterial cellulose composite.

7. The method according to claim 6, characterized in that The aqueous bacterial cellulose and the glycyrrhizic acid aqueous solution are each stirred to form a uniform solution.

8. The method according to claim 7, characterized in that The stirred aqueous bacterial cellulose and glycyrrhizic acid aqueous solution are each further subjected to ultrasonic treatment.

9. The method according to claim 6, characterized in that The dropwise addition is performed under ultrasound.

10. The method according to any one of claims 6 to 9, characterized in that The freeze-dried bacterial cellulose and water were mixed in a weight ratio of 1:10 to 15.

11. The method according to any one of claims 6 to 9, characterized in that The glycyrrhizic acid and water are mixed in a weight ratio of 1:0.5 to 1.5.

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