Glycyrrhizinate Bacterial Cellulose Complex for the Treatment of Skin Eczema

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 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 there is no background for its 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 nummular 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 keratinocytes 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 shortcomings of the prior art described above, the present invention provides a glycyrrhizic acid bacterial cellulose complex comprising bacterial cellulose supported with glycyrrhizic acid, wherein by topical application of the glycyrrhizic acid bacterial cellulose complex to the skin, it can effectively treat eczema, significantly alleviate the symptoms of eczema, be advantageous for the clinical treatment of skin eczema, and promote the repair of eczematous lesions, thereby providing a glycyrrhizic acid bacterial cellulose complex for treating skin eczema and / or alleviating the symptoms of eczema. [Means for solving the problem]

[0006] In one specific embodiment, the present invention provides the use of the glycyrrhizic acid bacterial cellulose complex for producing a drug that treats eczema and / or alleviates the symptoms of eczema.

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

[0008] In one specific embodiment, the bacterial cellulose is derived from 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 glycyrrhizic acid to bacterial cellulose is 1:0.1 to 10, preferably 1:5.

[0011] In one specific embodiment, the method for producing the glycyrrhizic acid bacterial cellulose complex 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 onto the water-containing bacterial cellulose to obtain the glycyrrhizic acid bacterial cellulose complex.

[0012] 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.

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

[0014] In one specific embodiment, the dripping is performed under ultrasonic conditions.

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

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

[0017] Specifically, the glycyrrhizic acid bacterial cellulose complex provided by the present invention is white and viscous, and glycyrrhizic acid is supported on its bacterial cellulose structure, enabling effective treatment of skin eczema.

[0018] The present invention further provides glycyrrhizic acid bacterial cellulose complexes in different proportions and compares the therapeutic effects of these complexes on skin eczema. As can be seen from the above, the glycyrrhizic acid bacterial cellulose complex provided by the present invention can significantly suppress the progression of lesions in skin where eczema occurs and can repair the lesioned area. The present invention obtains a substance that has a therapeutic effect on skin eczema by utilizing the amphiphilicity of glycyrrhizic acid and the biocompatibility, non-toxicity, and reticular structure of bacterial cellulose. Based on the research results on skin eczema treatment, the optimal doses of glycyrrhizic acid and bacterial cellulose are further selected in combination with pharmacopharmaceutical requirements to produce the relevant topical pharmaceutical formulation using the dosage ratio that provides the most favorable therapeutic effect on skin eczema. Since glycyrrhizic acid itself is an amphiphilic compound, and bacterial cellulose itself has characteristics such as non-toxicity and high biocompatibility, which are suitable for use as a carrier for skin care products, the present invention can be applied to research on the development of transdermal topical formulations that promote the treatment of eczema and the repair of eczematous skin. [Brief explanation of the drawing]

[0019] Embodiments of the present invention will be described with illustrative reference to the drawings. [Figure 1] SEM images of Comparative Examples 1, 3, and 5 are shown. [Figure 2] These are bar graphs showing the number of times rats scratched in comparative experiments 1-5 and experimental examples 1-3. [Figure 3] This shows the concentrations of different inflammatory indicators related to eczema in serum. [Figure 4] This shows the concentrations of different inflammatory indicators related to eczema in serum. [Figure 5] This shows the concentrations of different inflammatory indicators related to eczema in serum. [Figure 6] These are histological staining images of SD rat skin using hematoxylin-eosin (HE staining) from comparative experimental examples 1, 2, 5, and experimental example 3 (i.e., using the glycyrrhizic acid bacterial cellulose complex from Example 10).

Embodiments for Carrying Out the Invention

[0020] Hereinafter, 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 based on the content described in this specification. The present application can be implemented or applied by other different embodiments. As long as it does not conflict with the gist of the description of the present application, each detailed content of 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 can be combined. 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-inflammation and detoxification, 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 hydrophobic drugs, effectively increasing the solubility of the drugs and avoiding precipitation of the 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 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 properties of glycyrrhizic acid to develop and study bacterial cellulose as a topical pharmaceutical carrier, and finds that a 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 transport drug and bacterial cellulose as a carrier, and due to the amphiphilic properties of glycyrrhizic acid, glycyrrhizic acid is supported on the reticular structure of bacterial cellulose, creating 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 one specific embodiment, the glycyrrhizic acid is supported on bacterial cellulose to form a glycyrrhizic acid bacterial cellulose complex.

[0025] In one specific embodiment, the bacterial cellulose is made from Acetobacterium acetic acid bacteria. Derived from Bacteria, it has molecular weights of 50,000-2,500,000, 100,000-2,500,000, 500,000-2,500,000, 1,000,000-2,500,000, 1,500,000-2,500,000, 2,000,000-2,500,000, 50,000-2,000, 50,000-1,500,000, 50,000-1,000, 50,000-500,000, or 50,000-100,000. 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.

[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, 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.In other words, the bacterial cellulose is a high-molecular-weight polysaccharide composed of D-glucose linked by 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 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.

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

[0030] In one specific embodiment, the glycyrrhizic acid and water are mixed in 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 present application will be described in more detail below with reference to specific examples, but the scope of the present application is not limited by the description of the examples.

[0032] 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.

[0033] 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.

[0034] [Table 1]

[0035] 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.

[0036] 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.

[0037] 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.

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

[0039] To test the therapeutic effects of different samples on eczema, an eczema model was created using SD rats. Untreated SD rats (i.e., a blank group) were used as comparative experiment example 1. Then, the number of times the rats twisted their bodies after each sample was applied was statistically recorded, eczema-related inflammatory indicators were detected in the serum, and skin tissue was stained using HE. Details are as follows.

[0040] Comparative Experiment Example 2: DNCB-induced eczema model rats Prior to the experiment, a 3cm x 3cm area of ​​skin on the back of SD rats was depilated. On days 1 and 3 of the experiment, 2% dinitrochlorobenzene (DNCB) (Thermo Fisher Scientific) was applied to the depilated area, and on day 6, 0.5% DNCB was applied. Thereafter, 0.5% DNCB was applied once every three days (i.e., on days 9, 12, 15, and 18 of the experiment) to continuously stimulate the skin and obtain a DNCB-induced eczema model rat.

[0041] Comparative Experiment Example 3: Eczema Model Rats Treated with Glycyrrhizinic Acid Aqueous Solution Comparative Example 1 was applied to the eczema model rats obtained as described in Comparative Experiment Example 2 on the 7th day of the experiment.

[0042] Comparative Experiment Example 4: Eczema Model Rats Treated with Water-Containing Bacterial Cellulose The experiment was conducted in the same manner as in Comparative Experiment Example 3, except that Comparative Example 1, which was applied on the 7th day of the experiment, was replaced with Comparative Example 2.

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

[0044] Experimental Example 1: Eczema model rats treated with glycyrrhizic acid bacterial cellulose complex The experiment was conducted in the same manner as Comparative Experiment Example 3, except that Comparative Example 1, which was applied on the 7th day of the experiment, was replaced with Example 1.

[0045] Experimental Example 2: Eczema model rats treated with glycyrrhizic acid bacterial cellulose complex The experiment was conducted in the same manner as Comparative Experiment Example 3, except that Comparative Example 1, which was 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 The procedure was the same as in Comparative Experiment Example 3, except that Comparative Example 1, which was applied on the 7th day of the experiment, was replaced with Example 10.

[0047] On day 18 of the experiment, the number of times rats scratched (twitched) within a 20-minute period after the application of the sample was statistically recorded, and the results are shown in Figure 2. As shown in Figure 2, the application of the glycyrrhizic acid bacterial cellulose complex of Examples 1, 5, and 10 produced an antipruritic effect on eczematous areas of the skin, and may be equivalent to, or even superior to, conventional eczema treatment drugs (i.e., comparative example 5).

[0048] Nineteen days after the experiment, rats were sacrificed, and skin and serum from the depilated areas were collected and subjected to HE staining and detection of inflammatory markers, respectively. Referring to Figures 3-5, the concentrations of eczema-related inflammatory markers in serum detected by enzyme-linked immunosorbent assay (ELISA) shown in Figures 3-5 are those of immunoglobulin E (IgE), interleukin-4 (IL-4), and histamine, respectively. As shown in Figures 3-5, application of the glycyrrhizic acid bacterial cellulose complex of Examples 1, 5, and 10 significantly reduced the production of eczema inflammatory markers IgE, IL-4, and histamine. Figure 6 shows HE staining of SD rat skin, comparing the effects on the skin from the application of different samples. As shown in Figure 6, the skin area of ​​Example 3 to which the glycyrrhizic acid bacterial cellulose complex of Example 10 was applied showed a denser stratum corneum and increased epidermal thickness.

[0049] 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 treat skin eczema and alleviate its symptoms. 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.

[0050] 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. The use of a glycyrrhizic acid bacterial cellulose complex for manufacturing a drug for treating eczema and / or relieving the symptoms of eczema, wherein the glycyrrhizic acid bacterial cellulose complex comprises bacterial cellulose supported with glycyrrhizic acid, The aforementioned symptoms are inflammation.

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

4. The use 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 use according to claim 4, wherein the mass ratio of glycyrrhizic acid to bacterial cellulose is 1:

5.

6. The use according to any one of claims 1 to 3, wherein the method for producing the glycyrrhizic acid bacterial cellulose complex includes the following steps 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) 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 use described in 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 use according to claim 6, wherein the stirred water-containing bacterial cellulose and glycyrrhizic acid aqueous solution are each further treated with ultrasound.

9. The use according to claim 6, wherein the dropping is performed under ultrasonic conditions.

10. The use 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 use according to claim 6, wherein the glycyrrhizic acid and water are mixed in a weight ratio of 1:0.5 to 1.5.

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