Glycyrrhizinate bacterial cellulose complex promotes the 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 have 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 are few related studies on directly using bacterial cellulose as a drug carrier. Glycyrrhizic acid is one of the commonly used drugs in clinical practice and has effects such as anti-inflammatory and detoxifying, but glycyrrhizic acid has no background of actual application in topical pharmaceuticals.

[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. 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 promotes hair growth. A glycyrrhizic acid bacterial cellulose composite for promoting the growth and development of hair follicles and a method for producing the same are provided.

Means for Solving the Problems

[0005] In one specific embodiment, the bacterial cellulose is derived from 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, preferably 1:5.

[0008] Furthermore, the present invention provides a method for producing a glycyrrhizic acid bacterial cellulose complex, comprising 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 onto the water-containing bacterial cellulose to obtain the glycyrrhizic acid bacterial cellulose complex.

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

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

[0011] In one specific embodiment, the dripping is performed under ultrasonic conditions.

[0012] In one specific embodiment, the freeze-dried bacterial cellulose and water are mixed in a weight ratio of 1:10 to 15, preferably the glycyrrhizic acid and water are mixed in 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 complex is 1:0.1 to 10, preferably 1:5. [Effects of the Invention]

[0014] Specifically, the glycyrrhizic acid bacterial cellulose complex provided by the present invention is white and viscous, with glycyrrhizic acid supported on its 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 research results regarding the promotion of hair follicle growth and development in the skin, and in combination with pharmacopharmaceutical requirements, the optimal dosages of glycyrrhizic acid and bacterial cellulose can be further selected, and the relevant topical pharmaceutical formulations can be manufactured using the dosage ratio that provides the most favorable promotion effect on skin hair follicle growth and development. Because glycyrrhizic acid itself is an amphiphilic compound, and bacterial cellulose itself possesses characteristics such as non-toxicity and high biocompatibility, making it suitable for use as a carrier in skincare products, the present invention can be applied to research on the development of transdermal topical formulations for the treatment of hair loss and the promotion of hair growth. [Brief explanation of the drawing]

[0015] Embodiments of the present invention will be described with illustrative reference to the drawings. [Figure 1A] The SEM image of Comparative Example 1 is shown. [Figure 1B] The SEM image of Comparative Example 3 is shown. [Figure 1C] The SEM diagram of Example 5 is shown. [Figure 2] The glycyrrhizic acid content at different time points in Comparative Example 2, Example 1, Example 5, and Example 10 is shown. [Figure 3]The glycyrrhizic acid content at the skin penetration endpoint for Comparative Example 2, Example 1, Example 3, and Example 5 is shown. [Figure 4A] These are histological staining images of hair follicles in untreated and treated skin with the glycyrrhizic acid bacterial cellulose complex of Example 10. The skin was stained with toluidine blue and hematoxylin-eosin (HE) before penetration (left image) and at the penetration endpoint (right image). [Figure 4B] These are histological staining images of hair follicles in untreated and treated skin with the glycyrrhizic acid bacterial cellulose complex of Example 10. The skin was stained with toluidine blue and hematoxylin-eosin (HE) before penetration (left image) and at the penetration endpoint (right image). [Modes for carrying out the invention]

[0016] The embodiments of this application will be described below with reference to specific embodiments, but those skilled in the art will readily understand the advantages and effects of this application from the contents of this specification. This application can be implemented or applied by other different embodiments, and each detail of this specification can be modified and altered in different ways based on different perspectives and applications, as long as it does not contradict the spirit of this application. All ranges and numerical values ​​in this specification are comprehensive and can be combined. Any numerical value or endpoint that falls within the range described herein, for example, any integer, can be used to derive subranges, etc., as the minimum or maximum value.

[0017] Glycyrrhizic acid is a commonly used drug in clinical practice, possessing anti-inflammatory and detoxification properties. As a triterpene compound with amphiphilic properties, it exhibits characteristics as a surfactant. Glycyrrhizic acid aggregates or micelles can form host-guest inclusion complexes with hydrophobic drugs, effectively increasing drug solubility and avoiding drug precipitation.

[0018] 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 currently, there are few related studies on applying bacterial cellulose to topical 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 topical 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, a new structure is generated, and 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 made from Acetobacterium acetic acid bacteria. Derived from Balch), it has molecular weights 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 weights are 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 with 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, and 5,000 to 15,000 glucosyl groups. glucosyl groups, 6,000-15,000 glucosyl groups, 7,000-15,000 glucosyl groups, 8,000-15,000 glucosyl groups, 9,000-15,000 glucosyl groups, 10,000-15,000 glucosyl groups, 11,000-15,000 glucosyl groups, 12,000-15,000 glucosyl groups, 1 3,000-15,000 glucosyl groups, 14,000-15,000 glucosyl groups, 300-14,000 glucosyl groups, 300-13,000 glucosyl groups, 300-12,000 glucosyl groups, 300-11,000 glucosyl groups, 300-10,000 glucosyl groups, 300-9,000 glucosyl groups It has a syl group, 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 general chemical formula (C6H10O5)n and is a polysaccharide consisting of a linear chain (glycosidic bond) of hundreds to thousands of D-glucose units linked by β(1→4) bonds.That is, the bacterial cellulose is a high molecular polysaccharide composed of D-glucose with 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] Manufacturing example: Glycyrrhizinate bacterial cellulose complex Glycyrrhizic acid (93%, G810520-25g, Shanghai Macklin Biochemical Co.,Ltd.) was added to distilled water, mixed uniformly, and then subjected to sonication for 10 minutes to obtain an aqueous glycyrrhizic acid solution. Simultaneously, bacterial cellulose derived from acetic acid bacteria (nanobacteria cellulose freeze-dried tablets, EvoPhancie Biotech Ltd.) was added to distilled water, mixed uniformly, and then subjected to sonication for 10 minutes to obtain water-containing bacterial cellulose. Under sonication conditions, the aqueous glycyrrhizic acid solution was added dropwise to the water-containing bacterial cellulose at a rate of 5 drops per 10 seconds, mixed uniformly, and then subjected to sonication for 15 minutes to obtain a glycyrrhizic acid bacterial cellulose complex.

[0029] Using the method described in the above-mentioned manufacturing example, bacterial cellulose complexes of Examples 1 to 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 complex (product) is shown.

[0030] [Table 1]

[0031] Comparative Example 1: Aqueous solution of glycyrrhizic acid 10 mg of glycyrrhizic acid (93%, G810520-25 g, Shanghai Macklin Biochemical Co., Ltd.) was added to 1.6 mL of distilled water, mixed uniformly, and then subjected to sonication 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 (nanobacteria cellulose freeze-dried tablets, EvoPhancie Biotech Ltd.) was added to 1.3 mL of distilled water, mixed uniformly, and then subjected to sonication for 10 minutes to obtain water-containing bacterial cellulose.

[0033] Comparative Example 3: Baicalin Bacteria Cellulose 10 mg of baicalin (8802695-5g, Shanghai Macklin Biochemical Co., Ltd.) was added to 0.8 mL of distilled water, mixed uniformly, and then subjected to sonication for 10 minutes to obtain an aqueous baicalin solution. Simultaneously, 100 mg of bacterial cellulose derived from acetic acid bacteria (nanobacteria cellulose freeze-dried tablets, EvoPhancie Biotech Ltd.) was added to 1.3 mL of distilled water, mixed uniformly, and then subjected to sonication for 10 minutes to obtain water-containing bacterial cellulose. Under sonication conditions, the aqueous baicalin solution was added dropwise to the water-containing bacterial cellulose, mixed uniformly, and then subjected to sonication for 15 minutes. After rotational evaporation, a baicalin bacterial cellulose complex was obtained.

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

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

[0036] Comparative Example 2, Example 1, Example 5, and Example 10 were each used as samples for the skin penetration experiment and high-performance liquid chromatography. For each sample, the measurement was repeated three times and the average value was calculated, and the results are shown in Figure 2. Furthermore, Comparative Example 2, Example 1, Example 3, and Example 5 were each used as samples for the skin penetration experiment and high-performance liquid chromatography, and the glycyrrhizic acid content at the skin penetration endpoint (i.e., 360 minutes after the start of the experiment) was measured. The results shown in Figures 2 and 3 indicate that glycyrrhizic acid was not detected in the absorption solution of Comparative Example 2. This suggests that the cumulative penetration amount of glycyrrhizic acid increases with increasing proportion of bacterial cellulose in the glycyrrhizic acid bacterial cellulose complex, indicating that glycyrrhizic acid in a glycyrrhizic acid bacterial cellulose complex with a high proportion of bacterial cellulose has enhanced skin permeability.

[0037] Referring to Figures 4A and 4B, the skin was stained with toluidine blue (left figure) and HE staining (right figure) before the skin penetration experiment and upon reaching the skin penetration endpoint, respectively. The untreated skin tissue and the skin tissue treated with the glycyrrhizic acid bacterial cellulose complex of Example 10 were compared, and the changes in the number and morphology of hair follicles in an equivalent area of ​​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 contains 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. Furthermore, the glycyrrhizic acid bacterial cellulose complex provided by the present invention has a simple manufacturing process, and the ratio of glycyrrhizic acid and bacterial cellulose contained in the complex can be easily adjusted, giving it promising application prospects.

[0039] The above embodiments are illustrative and not limiting to the present application. Those skilled in the art can modify and alter the above embodiments without departing from the spirit and scope of the present application. Accordingly, the claims of the present application are defined by the claims appended to this specification and are included in the technical content of this publication, provided that they do not affect the effects and purposes of the present application.

Claims

1. A glycyrrhizic acid bacterial cellulose complex for application to the skin to promote the growth and development of skin hair follicles, comprising glycyrrhizic acid and bacterial cellulose, characterized in that the glycyrrhizic acid is supported on a reticular structure of bacterial cellulose.

2. The glycyrrhizic acid bacterial cellulose complex according to claim 1, wherein the bacterial cellulose is derived from Acetobacterium balch and has a molecular weight of 50,000 to 2,500,000.

3. The bacterial cellulose has 300 to 15,000 glucosyl groups, as described in claim 1, the glycyrrhizic acid bacterial cellulose complex.

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

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

5.

6. A method for producing a glycyrrhizic acid bacterial cellulose complex according to any one of claims 1 to 3, 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 solution of glycyrrhizic acid, 3) A method comprising the step of adding the aqueous glycyrrhizic acid solution dropwise to the water-containing bacterial cellulose to obtain the glycyrrhizic acid bacterial cellulose complex.

7. The method according to claim 6, wherein the aqueous solution containing water-containing bacterial cellulose and the aqueous solution of glycyrrhizic acid 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 according to claim 6, wherein the dropping is performed under ultrasonic conditions.

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