Glycyrrhizic acid bacterial cellulose composite for treating eczema in skin

Abstract

To provide the use of a composite for treatment of eczema.SOLUTION: The present invention provides the use of a glycyrrhizic acid bacterial cellulose composite for producing a drug for treating eczema and / or alleviating the symptom of eczema, the glycyrrhizic acid bacterial cellulose composite including the bacterial cellulose loaded with the glycyrrhizic acid.SELECTED DRAWING: Figure 6

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, easy 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 effects, but glycyrrhizic acid has no background of actual application in topical pharmaceuticals.

[0003] Eczema (i.e., atopic dermatitis or atopic eczema) is the largest burden of skin disease disorders worldwide, affecting nearly 20% of children worldwide. Eczema can exist in various forms, including atopic and non-atopic forms. The atopic form is usually mediated by IgE, and the non-atopic form is mediated by non-IgE. Both types of forms can be manifested as an increase in eosinophils. Other variants of eczema include nodular eczema, seborrheic dermatitis, and hand eczema. Eczema can also be manifested and appear as a skin disease of other systemic diseases, such as Wiskott-Aldrich syndrome, human immunodeficiency virus (HIV) infection, or food allergy. In the process of eczema onset, excessive proliferation of keratin-producing cells and excessive expression of inflammatory factors are prominent.

[0004] Therefore, the clinical treatment of eczema is currently a difficult problem that the industry wants to overcome.

Summary of the Invention

Problems to be Solved by the Invention

[0005] In view of the various defects of the above prior art, the present invention provides a glycyrrhizic acid bacterial cellulose composite containing glycyrrhizic acid-supported bacterial cellulose. By topically applying the glycyrrhizic acid bacterial cellulose composite to the skin, eczema can be effectively treated, the symptoms of eczema can be significantly alleviated, which is advantageous for the clinical treatment of skin eczema and promotes the repair of the lesion site of eczema. The present invention provides a glycyrrhizic acid bacterial cellulose composite for treating skin eczema and / or alleviating the symptoms of eczema.

Means for Solving the Problems

[0006] In one specific embodiment, the present invention provides the use of the glycyrrhizic acid bacterial cellulose composite for manufacturing a drug for treating eczema and / or alleviating the symptoms of eczema.

[0007] In one specific embodiment, the symptoms include erythema, edema, itching or any combination thereof.

[0008] 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.

[0009] In one specific embodiment, the bacterial cellulose has 300 to 15,000 glucosyl groups.

[0010] In one specific embodiment, the mass ratio of the glycyrrhizic acid to the bacterial cellulose is 1:0.1 to 10, preferably, the mass ratio of the glycyrrhizic acid to the bacterial cellulose is 1:5.

[0011] In one specific embodiment, the method for producing the glycyrrhizic acid bacterial cellulose composite comprises 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.

[0012] In one specific embodiment, the water-containing bacterial cellulose and the aqueous glycyrrhizic acid solution are each stirred to form a uniform solution.

[0013] In one specific embodiment, the stirred water-containing bacterial cellulose and the aqueous glycyrrhizic acid solution are each further treated with ultrasonic waves.

[0014] In one specific embodiment, the dropping is performed under ultrasonic waves.

[0015] In one specific embodiment, the freeze-dried bacterial cellulose and water are mixed at a weight ratio of 1:10 to 15.

[0016] In one specific embodiment, the glycyrrhizic acid and water are mixed at a weight ratio of 1:0.5 to 1.5.

Advantages of the Invention

[0017] Specifically, the glycyrrhizic acid bacterial cellulose composite provided by the present invention is white and viscous, and the bacterial cellulose structure carries glycyrrhizic acid, which can effectively treat skin eczema.

[0018] The present invention further provides a glycyrrhizic acid-bacterial cellulose composite having different ratios, and compared the therapeutic effects of the composite on skin eczema. As can be seen from the above content, the glycyrrhizic acid-bacterial cellulose composite provided by the present invention can significantly inhibit the progression of lesions in the skin where eczema occurs and repair the lesion sites. The present invention utilizes the amphiphilicity of glycyrrhizic acid and the biocompatibility, non-toxicity and network structure of bacterial cellulose to obtain a substance having a therapeutic effect on skin eczema. Based on the research results of skin eczema treatment, in combination with the requirements of pharmaceutical formulations, the optimal dosages of glycyrrhizic acid and bacterial cellulose are further selected, and the most preferable administration ratio of the therapeutic effect on skin eczema is utilized to manufacture related topical pharmaceutical formulations. 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 transdermal topical preparations for promoting the treatment of eczema and the repair of eczema skin.

Brief Description of the Drawings

[0019] The embodiments of the present application will be described by referring to the drawings by way of example.

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Best Mode for Carrying Out the Invention

[0020] Hereinafter, embodiments of the present application will be described by 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 gist 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.

[0021] Glycyrrhizic acid is a drug commonly used clinically, has effects such as anti-inflammatory and detoxifying, is a triterpenoid compound, and has amphiphilicity, so it exhibits 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.

[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 currently there are few related studies on applying bacterial cellulose to topical pharmaceutical carriers including promoting hair growth.

[0023] Therefore, the present invention utilizes the amphiphilic property 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, bacterial cellulose as a carrier, and due to the amphiphilic property 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.

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

[0025] 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.

[0026] 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 with β-1,4-glycosidic bonds.

[0027] 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.

[0028] 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.

[0029] 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.

[0030] 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.

[0031] 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.

[0032] Production Example: Glycyrrhizic Acid - Bacterial Cellulose Composite Glycyrrhizic acid (93%, G810520 - 25g, Shanghai Macklin Biochemical Co., Ltd.) was added to distilled water. After uniformly stirring, 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. After uniformly stirring, 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. After uniformly stirring, ultrasonic treatment was carried out for 15 minutes to obtain a glycyrrhizic acid - bacterial cellulose composite.

[0033] By the method of the above production example, bacterial cellulose composites of Examples 1 - 19 were produced with the compositions shown in Table 1 below, and the mass ratios of glycyrrhizic acid to bacterial cellulose in the produced glycyrrhizic acid - bacterial cellulose composites (products) are shown.

[0034] [Table 1]

[0035] 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. After uniformly stirring, ultrasonic treatment was carried out for 10 minutes to obtain an aqueous solution of glycyrrhizic acid.

[0036] 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. After uniformly stirring, ultrasonic treatment was carried out for 10 minutes to obtain water - containing bacterial cellulose.

[0037] 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. After uniformly stirring, ultrasonic treatment was performed for 10 minutes to obtain a bikaverin aqueous 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. After uniformly stirring, ultrasonic treatment was performed for 10 minutes to obtain water-containing bacterial cellulose. Under ultrasonic conditions, the bikaverin aqueous solution was dropped into the water-containing bacterial cellulose. After uniformly stirring, ultrasonic treatment was performed for 15 minutes, and then rotary evaporation was carried out to obtain a bikaverin bacterial cellulose composite.

[0038] Referring to Figure 1, Comparative Example 1, Comparative Example 3, and Comparative 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 Figure 1. As shown in Figure 1, 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.

[0039] To test the therapeutic effects of different samples on eczema, an eczema model was established using SD rats. SD rats without any treatment (i.e., the blank group) were used as Comparative Experimental Example 1. Next, the number of times the rats twisted their bodies after applying each sample was counted, the eczema-related inflammatory indicators in the serum were detected, and the skin tissue was stained with HE. The details are as follows.

[0040] Comparative Experimental Example 2: DNCB-induced Eczema Model Rats Before the experiment, the skin area of 3 cm×3 cm on the back of SD rats was depilated in advance. On the 1st and 3rd days after the start of the experiment, 2% dinitrochlorobenzene (DNCB) (Thermo Fisher Scientific) was applied to the depilated area respectively. On the 6th day of the experiment, 0.5% DNCB was applied, and then, every 3 days (i.e., on the 9th, 12th, 15th and 18th days of the experiment), 0.5% DNCB was applied once to continuously stimulate the skin and obtain DNCB-induced eczema model rats.

[0041] Comparative Experimental Example 3: Eczema model rats treated with aqueous glycyrrhizinate solution On the 7th day of the experiment, Comparative Example 1 was applied to the eczema model rats obtained as in the previous Comparative Experimental Example 2.

[0042] Comparative Experimental Example 4: Eczema model rats treated with water-containing bacterial cellulose It was carried out in the same manner as in the previous Comparative Experimental Example 3, except that Comparative Example 1 applied on the 7th day of the experiment was replaced with Comparative Example 2.

[0043] Comparative Experimental Example 5: Eczema model rats treated with mometasone furoate cream It was carried out in the same manner as in the previous Comparative Experimental Example 3, except that Comparative Example 1 applied on the 7th day of the experiment was replaced with mometasone furoate cream (0.1% (5 g: 5 mg), 220908 Bayer Healthcare (Shanghai) Ltd.).

[0044] Experimental Example 1: Eczema model rats treated with glycyrrhizinate-bacterial cellulose complex It was carried out in the same manner as in the previous Comparative Experimental Example 3, except that Comparative Example 1 applied on the 7th day of the experiment was replaced with Example 1.

[0045] Experimental Example 2: Eczema model rats treated with glycyrrhizinate-bacterial cellulose complex It was carried out in the same manner as in the previous Comparative Experimental Example 3, except that Comparative Example 1 applied on the 7th day of the experiment was replaced with Example 5.

[0046] Experimental Example 3: Eczema model rats treated with glycyrrhizic acid - bacterial cellulose complex Comparative Example 1 applied on the 7th day of the experiment was replaced with Example 10, and the rest was the same as Comparative Experimental Example 3.

[0047] For 30 minutes after applying the sample on the 18th day of the experiment, the number of times the rats scratched (twisted their bodies) within 20 minutes was counted, and the results are shown in Figure 2. As shown in Figure 2, when the glycyrrhizic acid - bacterial cellulose complexes of Examples 1, 5, and 10 were applied, an antipruritic effect was exerted on the eczema area of the skin, and the effect could be equivalent to or even better than that of the conventional eczema treatment drug (i.e., Comparative Experimental Example 5).

[0048] On the 19th day after the experiment, the rats were sacrificed, and the skin and serum of the depilated area were collected, and HE staining and detection of inflammatory indicators were performed respectively. Referring to Figures 3 - 5, the concentrations of eczema - related inflammatory indicators in the serum detected by enzyme - linked immunosorbent assay (ELISA) shown in Figures 3 - 5 are the concentrations of immunoglobulin E (IgE), interleukin - 4 (IL - 4), and histamine respectively. As shown in Figures 3 - 5, when the glycyrrhizic acid - bacterial cellulose complexes of Examples 1, 5, and 10 were applied, the generation of eczema inflammatory indicators IgE, IL - 4, and histamine could be significantly reduced. Also, Figure 6 shows the HE staining diagram of the skin of SD rats, comparing the effects on the skin by applying different samples. As shown in Figure 6, in the skin area of Example 3 where the glycyrrhizic acid - bacterial cellulose complex of Example 10 was applied, the skin layer was dense and the thickness of the epidermal layer increased.

[0049] 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, can effectively treat skin eczema and relieve its symptoms. Also, the glycyrrhizic acid - bacterial cellulose complex provided by the present invention has a simple manufacturing process, can easily adjust the ratio of glycyrrhizic acid and bacterial cellulose contained in the complex, and has application prospects.

[0050] 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. 1. Use of a glycyrrhizinic acid-bacterial cellulose complex for the manufacture of a medicament for treating eczema and / or alleviating the symptoms of eczema, the glycyrrhizinic acid-bacterial cellulose complex comprising bacterial cellulose carrying glycyrrhizinic acid.

2. The use of claim 1 , wherein the symptoms include redness, swelling, itching, or any combination thereof.

3. 2. The use 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.

4. The use according to claim 1, wherein the bacterial cellulose has 300 to 15,000 glucosyl groups.

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

6. The use according to claim 5, wherein the mass ratio of glycyrrhizinic acid to bacterial cellulose is 1:

5.

7. The use according to any one of claims 1 to 4, wherein the method for producing the glycyrrhizinic acid bacterial cellulose complex comprises the following steps 1) to 3): 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) A step of dropping the glycyrrhizinic acid aqueous solution onto the water-containing bacterial cellulose to obtain the glycyrrhizinic acid bacterial cellulose complex.

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

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

10. The use according to claim 7, wherein the instillation is carried out under ultrasound.

11. The use according to claim 7, wherein the freeze-dried bacterial cellulose and water are mixed in a weight ratio of 1:10-15.

12. The use according to claim 7, wherein the glycyrrhizic acid and water are mixed in a weight ratio of 1:0.5 to 1.5.

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