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

Abstract

To provide a compound useful for clinical treatment of alopecia, and a method for producing the same.SOLUTION: There is provided a glycyrrhizic acid bacterial cellulose composite for promoting the growth and development of skin hair follicles, including the glycyrrhizic acid and the bacterial cellulose.SELECTED DRAWING: Figure 1C

Description

Technical Field

[0001] The present invention relates to a bacterial cellulose composite, particularly a glycyrrhizic acid bacterial cellulose composite, and a method for producing the same.

Background Art

[0002] Bacterial cellulose is a relatively new nanomaterial with few applications in drug delivery. It has been proven to simultaneously possess various special physical and chemical properties, including biodegradability, non-toxicity, high elastic modulus, high specific surface area, low density, non-abrasiveness, ease of surface functionalization, high purity and crystallinity in chemical composition, high degree of polymerization (2000 - 8000), and high hardness. Also, since bacterial cellulose is an inert material, there is little related research on directly using bacterial cellulose as a drug carrier. Glycyrrhizic acid is one of the drugs commonly used clinically and has effects such as anti-inflammatory and detoxifying, but there is no background for its actual application as a topical pharmaceutical.

[0003] Therefore, the clinical treatment of hair loss is currently a problem that the industry wants to overcome.

Summary of the Invention

Problems to be Solved by the Invention

[0004] In view of the various defects of the above prior art, the present invention provides a glycyrrhizic acid bacterial cellulose composite containing glycyrrhizic acid and bacterial cellulose, and by locally applying the glycyrrhizic acid bacterial cellulose composite to the skin, it can promote the growth and development of hair follicles in the skin, which is advantageous for the clinical treatment of hair loss, and provides a glycyrrhizic acid bacterial cellulose composite that promotes hair growth and promotes the growth and development of hair follicles, and a method for producing the same.

Means for Solving the Problems

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

[0006] In one specific embodiment, the bacterial cellulose has 300 to 15,000 glucosyl groups.

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

[0008] Furthermore, the present invention provides a method for producing the glycyrrhizic acid-bacterial cellulose composite, which includes the steps of: 1) mixing water and freeze-dried bacterial cellulose to obtain water-containing bacterial cellulose; 2) dissolving glycyrrhizic acid in water to obtain an aqueous glycyrrhizic acid solution; and 3) dropping the aqueous glycyrrhizic acid solution into the water-containing bacterial cellulose to obtain the glycyrrhizic acid-bacterial cellulose composite.

[0009] In one specific embodiment, the water-containing bacterial cellulose and the aqueous glycyrrhizic acid solution are each stirred to form a uniform solution.

[0010] In one specific embodiment, the stirred water-containing bacterial cellulose and the aqueous glycyrrhizic acid solution are each further treated with ultrasonic waves.

[0011] In one specific embodiment, the dropping is performed under ultrasonic waves.

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

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

Advantages of the Invention

[0014] Specifically, the glycyrrhizic acid - bacterial cellulose composite provided by the present invention is white and viscous, and glycyrrhizic acid is carried on the bacterial cellulose structure. 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. Based on the research results of the promoting effect on the growth and development of hair follicles in the skin, in combination with the requirements of pharmaceutical formulation science, the optimal dosages of glycyrrhizic acid and bacterial cellulose are further selected, and the most preferable administration ratio for promoting the growth and development of skin hair follicles is utilized to manufacture related topical pharmaceutical preparations. Since glycyrrhizic acid itself is an amphiphilic compound, and the characteristics of bacterial cellulose itself include non - toxicity and high biocompatibility, and there is a background for use as a skin - care product carrier, the present invention can be applied to the development research of treatments for hair removal and transdermal topical preparations for promoting hair growth.

Brief Description of the Drawings

[0015] The embodiments of the present application will be described by referring to the drawings exemplarily.

Figure 1A

Figure 1B

Figure 1C

Figure 2

Figure 3

Figure 4A

Figure 4B

Embodiments for Carrying Out the Invention

[0016] Hereinafter, the embodiments of the present application will be described with specific specific embodiments. Those skilled in the art can easily understand the advantages and effects of the present application from the description in this specification. The present application can be implemented or applied by other different embodiments. As long as it does not violate the spirit of the description of the present application, each detailed content in this specification can be modified and changed based on different viewpoints and applications. Note that all ranges and numerical values in this specification are inclusive and combinable. Any numerical value or endpoint within the range described in this specification, for example, any integer, can be used as the minimum value or the maximum value to derive sub-ranges and the like.

[0017] Glycyrrhizic acid is a drug commonly used clinically, has effects such as anti-inflammatory and detoxifying effects, is a triterpenoid compound, and has amphiphilic properties, so it exhibits the characteristics as a surfactant. Aggregates or micelles of glycyrrhizic acid can form an inclusion complex, which is a host-guest body, with a hydrophobic drug, effectively increasing the solubility of the drug and avoiding precipitation of the drug.

[0018] Bacterial cellulose has characteristics such as biodegradability, non-toxicity, high elastic modulus, high specific surface area, low density, non-abrasiveness, ease of 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 currently, there are few related studies on applying bacterial cellulose to local pharmaceutical carriers including promoting hair growth.

[0019] Therefore, the present invention utilizes the amphiphilicity of glycyrrhizic acid to develop and study bacterial cellulose as a local pharmaceutical carrier, and finds 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 a drug to be transported and bacterial cellulose as a carrier. Due to the amphiphilicity of glycyrrhizic acid, glycyrrhizic acid is carried on the network structure of bacterial cellulose, generating a new structure. The formed glycyrrhizic acid - bacterial cellulose complex can significantly promote the growth and development of hair follicles in the skin.

[0020] In one specific embodiment, the glycyrrhizic acid is carried on the bacterial cellulose to form a glycyrrhizic acid - bacterial cellulose complex.

[0021] In one specific embodiment, the bacterial cellulose is derived from acetic acid bacteria (Acetobacterium Balch) and has a molecular weight of 50,000 to 2,500,000, 100,000 to 2,500,000, 500,000 to 2,500,000, 1,000,000 to 2,500,000, 1,500,000 to 2,500,000, 2,000,000 to 2,500,000, 50,000 to 2,000,000, 50,000 to 1,500,000, 50,000 to 1,000,000, 50,000 to 500,000 or 50,000 to 100,000. For example, the molecular weight is 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 acetic acid bacteria.

[0022] In one 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, it has 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 a chemical general formula of (C6H10O5)n and is a polysaccharide composed of a linear chain (glycosidic bond) of several hundred to several thousand D-glucose units linked by β(1→4).That is, the bacterial cellulose is a high-molecular polysaccharide composed of D-glucose linked by β-1,4-glycosidic bonds.

[0023] In one specific embodiment, the mass ratio of glycyrrhizic acid to bacterial cellulose is 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.

[0024] The method for producing the glycyrrhizic acid-bacterial cellulose composite provided by the present invention includes: 1) a step of mixing water and freeze-dried bacterial cellulose to obtain water-containing bacterial cellulose; 2) a step of dissolving glycyrrhizic acid in water to obtain an aqueous glycyrrhizic acid solution; and 3) a step of dropping the aqueous glycyrrhizic acid solution into the water-containing bacterial cellulose to obtain the glycyrrhizic acid-bacterial cellulose composite.

[0025] In one 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.

[0026] In one specific embodiment, glycyrrhizic acid and water are mixed at a weight ratio of 1:0.5 to 1.5, for example, a weight ratio of 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.

Examples

[0027] Hereinafter, the present application will be described in more detail with specific examples, but the scope of the present application is not limited by the description of the examples.

[0028] Production Example: Glycyrrhizic Acid - Bacterial Cellulose Composite Glycyrrhizic acid (93%, G810520 - 25g, Shanghai Macklin Biochemical Co., Ltd.) was added to distilled water and stirred uniformly. Then, ultrasonic treatment was carried out for 10 minutes to obtain an aqueous solution of glycyrrhizic acid. At the same time, bacterial cellulose derived from acetic acid bacteria (nanobacterial cellulose freeze - dried tablets, EvoPhancie Biotech Ltd.) was added to distilled water, stirred uniformly, and then ultrasonic treatment was carried out for 10 minutes to obtain water - containing bacterial cellulose. Under ultrasonic conditions, the aqueous solution of glycyrrhizic acid was dropped into the water - containing bacterial cellulose at a rate of 5 drops per 10 seconds, stirred uniformly, and then ultrasonic treatment was carried out for 15 minutes to obtain a glycyrrhizic acid - bacterial cellulose composite.

[0029] By the method of the above - mentioned production example, bacterial cellulose composites of Examples 1 - 19 were produced with the compositions shown in Table 1 below, and the mass ratio of glycyrrhizic acid to bacterial cellulose in the produced glycyrrhizic acid - bacterial cellulose composites (products) is shown.

[0030]

Table 1

[0031] Comparative Example 1: Aqueous Solution of Glycyrrhizic Acid 10 mg of glycyrrhizic acid (93%, G810520 - 25g, Shanghai Macklin Biochemical Co., Ltd.) was added to 1.6 mL of distilled water, stirred uniformly, and then ultrasonic treatment was carried out for 10 minutes to obtain an aqueous solution of glycyrrhizic acid.

[0032] Comparative Example 2: Water - containing Bacterial Cellulose 50 mg of bacterial cellulose derived from acetic acid bacteria (nanobacterial cellulose freeze - dried tablets, EvoPhancie Biotech Ltd.) was added to 1.3 mL of distilled water, stirred uniformly, and then ultrasonic treatment was carried out for 10 minutes to obtain water - containing bacterial cellulose.

[0033] Comparative Example 3: Bikaverin Bacterial Cellulose 10 mg of bikaverin (8802695-5g, Shanghai Macklin Biochemical Co., Ltd.) was added to 0.8 mL of distilled water and stirred uniformly. After that, ultrasonic treatment was performed for 10 minutes to obtain an aqueous bikaverin solution. At the same time, 100 mg of bacterial cellulose derived from acetic acid bacteria (nanobacterial cellulose freeze-dried tablets, EvoPhancie Biotech Ltd.) was added to 1.3 mL of distilled water, stirred uniformly, and then ultrasonic treatment was performed for 10 minutes to obtain water-containing bacterial cellulose. Under ultrasonic conditions, the aqueous bikaverin solution was dropped into the water-containing bacterial cellulose, stirred uniformly, and then ultrasonic treatment was performed for 15 minutes. After rotary evaporation, a bikaverin bacterial cellulose composite was obtained.

[0034] Referring to FIGS. 1A to 1C, Comparative Example 1, Comparative Example 3, and Example 5 were each analyzed with a scanning electron microscope (FEI Quanta 400 FEI, America FEI scanning electron microscope), and the obtained SEM images are shown in FIGS. 1A to 1C respectively. As shown in FIGS. 1A to 1C, only the glycyrrhizic acid bacterial cellulose composite obtained in Example 5 has a structure in which glycyrrhizic acid is wrapped in the network structure of bacterial cellulose.

[0035] To show the ability of different samples of glycyrrhizic acid to penetrate the skin (i.e., Comparative Examples 1 to 3 and Examples 1 to 19), a skin penetration experiment was conducted using a Franz cell diffusion cell with the skin on the back of SD rats. First, before the experiment, the skin (3 cm × 3 cm) in the diffusion cell was washed with 50% ethanol, and 200 mg of the sample was uniformly applied to the washed skin. After the experiment was completed, the skin was washed with 20% ethanol to remove the surface glycyrrhizic acid. At 2, 4, and 6 hours later, the absorption solution was collected respectively, and 0.1 mL of the absorption solution was mixed with 0.9 mL of methanol. Next, high-performance liquid chromatography was performed on the mixed solution of the absorption solution and methanol to measure the concentration of glycyrrhizic acid in the skin at different time points.

[0036] Using Comparative Example 2, Example 1, Example 5, and Example 10 as samples respectively, the skin penetration experiment and high-performance liquid chromatography were carried out. For each sample, the measurement was repeated 3 times and the average value was calculated, and the results are shown in Figure 2. In addition, using Comparative Example 2, Example 1, Example 3, and Example 5 as samples respectively, the skin penetration experiment and high-performance liquid chromatography were carried out, and the content of glycyrrhizic acid at the skin penetration end point (that is, 360 minutes after the start of the experiment) was measured. From the results shown in Figures 2 and 3, the presence of glycyrrhizic acid has not been detected in the absorption solution of Comparative Example 2, and with the increase in the proportion of bacterial cellulose in the glycyrrhizic acid-bacterial cellulose complex, the cumulative penetration amount of glycyrrhizic acid increases, indicating that the glycyrrhizic acid in the glycyrrhizic acid-bacterial cellulose complex having a high proportion of bacterial cellulose has enhanced skin permeation ability.

[0037] Referring to Figures 4A and 4B, before the skin penetration experiment and when the skin penetration end point was reached, the skin was stained with toluidine blue (left figure) and HE staining (right figure) respectively, and the skin tissues untreated and treated with the glycyrrhizic acid-bacterial cellulose complex of Example 10 were compared, and the number and morphological changes in the hair follicles in the equivalent area skin were observed. As shown in Figures 4A and 4B, the glycyrrhizic acid-bacterial cellulose complex provided by the present invention can promote the growth and development of hair follicles in the skin.

[0038] As described above, 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 glycyrrhizic acid-bacterial cellulose complex provided by the present invention has a simple manufacturing process, can easily adjust the proportion of glycyrrhizic acid and bacterial cellulose contained in the complex, and has application prospects.

[0039] The above embodiments are described by way of example and are not intended to limit the present application. Those skilled in the art can make modifications and changes to 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 claims attached hereto, and includes the technical content disclosed herein as long as it does not affect the effects and implementation purposes of the present application.

Claims

1. A glycyrrhizinic acid-bacterial cellulose complex containing glycyrrhizinic acid and bacterial cellulose, which promotes the growth and development of skin hair follicles.

2. 2. The glycyrrhizinic acid bacterial cellulose complex according to claim 1, wherein the bacterial cellulose is derived from acetic acid bacteria (Acetobacterium Balch) and has a molecular weight of 50,000 to 2,500,000.

3. The glycyrrhizinic acid bacterial cellulose complex according to claim 1, wherein the bacterial cellulose has 300 to 15,000 glucosyl groups.

4. The glycyrrhizinic acid bacterial cellulose complex according to any one of claims 1 to 3, wherein the mass ratio of glycyrrhizinic acid to bacterial cellulose is 1:0.1-10.

5. The glycyrrhizinic acid bacterial cellulose complex according to any one of claims 1 to 3, wherein the mass ratio of the glycyrrhizinic acid to the bacterial cellulose is 1:

5.

6. A method for producing the glycyrrhizinic acid bacterial cellulose complex according to any one of claims 1 to 3, comprising the steps of: 1) mixing water and freeze-dried bacterial cellulose to obtain water-containing bacterial cellulose; 2) dissolving glycyrrhizinic acid in water to obtain an aqueous solution of glycyrrhizinic acid; and 3) adding the aqueous glycyrrhizinic acid solution dropwise to the water-containing bacterial cellulose to obtain the glycyrrhizinic acid bacterial cellulose complex.

7. The method according to claim 6, wherein the water-containing bacterial cellulose and the aqueous glycyrrhizic acid solution are each stirred to form a homogeneous solution.

8. The method according to claim 7, wherein the stirred water-containing bacterial cellulose and glycyrrhizic acid aqueous solution are each further treated with ultrasound.

9. The method of claim 6 , wherein the dropping is performed under ultrasound.

10. The method according to claim 6, wherein the freeze-dried bacterial cellulose and water are mixed in a weight ratio of 1:10-15.

11. The method according to claim 6, wherein the freeze-dried bacterial cellulose and water are mixed in a weight ratio of 1:0.5 to 1.5.

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