Lucid ganoderma-derived protein oral soluble film as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a lucid ganoderma-derived protein oral soluble film as well as a preparation method and application thereof. The protein oral soluble film derived from the lucid ganoderma is prepared from the following raw materials in percentage by mass: 1 to 10 percent of GMI (Microspore Ganoderma Immunoprotein), 25 to 50 percent of a film forming material, 5 to 20 percent of a plasticizer, 5 to 30 percent of a mucous membrane adhesive, 1 to 20 percent of a penetration enhancer, 5 to 20 percent of a flavoring agent, 0.1 to 1 percent of a coloring agent and 0.5 to 3 percent of an acidity regulator. According to the invention, by optimizing the types and proportions of the components in the formula, the oral soluble film not only has excellent appearance quality, but also has relatively good folding resistance and relatively short disintegration time, so that the use experience of the product is further improved.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations, and in particular relates to a ganoderma-derived protein oral dissolving film and a preparation method and application thereof. Background Art

[0002] A protein derived from Ganoderma lucidum: Ganoderma microsporum immunomodulatory protein (GMI) is derived from Ganoderma microsporum and is a fungal immunomodulatory protein with multiple biological activities. It has a wide range of biological activities and potential clinical application value, especially in the fields of anti-inflammation, promoting mucosal healing, immunomodulation and antiviral.

[0003] At present, the main method of intravenous injection is to inject protein and peptide active ingredients in clinical applications. Although this method of administration can directly enter the blood circulation to ensure the biological activity of the drug, it also has significant disadvantages, including poor patient compliance, complex operation, high production cost and inconvenience in use, especially for patients who need long-term medication. Therefore, the development of innovative preparations that can replace injection has become a research focus. As a new drug delivery system, oral dissolving film can quickly dissolve and release active ingredients in the oral cavity, so as to be absorbed through the oral mucosa, sublingual vein or gastrointestinal tract. Compared with the traditional injection method, oral dissolving film has significant advantages, such as waterless administration, dissolving in the mouth, and easy use, which is particularly suitable for improving patient compliance. In addition, the preparation process of oral dissolving film can achieve precise dosage control, and has good stability and portability. However, for protein and peptide macromolecules, delivery through oral dissolving film still faces many challenges. Proteins and peptides have complex structures and large molecular weights. They are easily enzymatically hydrolyzed and destroyed by gastric acid in the gastrointestinal tract, and there is an intestinal absorption barrier, resulting in low bioavailability. Existing technologies are mostly concentrated on the development of orally dissolving films for small molecule compounds, while there are few breakthroughs in the application of large molecules such as polypeptide proteins. The technical barriers are mainly reflected in the protection of active ingredient stability, improvement of absorption efficiency and optimization of formulation processes.

[0004] For example, Chinese patent CN107080745A discloses an oral fast-dissolving film containing lutein albumin nanoparticles and a preparation method thereof. Lutein is first prepared into lutein albumin nanoparticles by an anti-solvent precipitation method, and then a solvent casting method is used to prepare an oral fast-dissolving film containing lutein albumin nanoparticles. The film has a complete surface without bubbles, uniform color, consistent thickness, and uniform drug distribution. Compared with the lutein raw material and the oral fast-dissolving film prepared therefrom, the film has significantly increased in vitro release and in vivo absorption speed and degree, and significantly improved bioavailability.

[0005] For another example, patent CN112823808B discloses a pharmaceutical composition for treating epithelial cell cancer and its use. The pharmaceutical composition comprises microspore Ganoderma lucidum immunomodulatory protein and keyhole limpet hemocyanin. The epithelial cell cancer is lung cancer, breast cancer, oral cancer, esophageal cancer, gastric cancer, colorectal cancer, liver cancer, cervical cancer, bladder cancer, pancreatic cancer or skin cancer. The composition can enhance the immunogenicity or therapeutic effect of cancer vaccines, but the dosage form does not have an orodispersible film, and its application is limited to medicine.

[0006] Therefore, there is an urgent need to develop a technology that can combine protein and polypeptide preparation technology with oral film technology to overcome the barriers of existing technologies. Summary of the invention

[0007] Based on the shortcomings of the prior art, the present invention aims to provide a protein oral dissolving film derived from Ganoderma lucidum and a preparation method thereof. The protein oral dissolving film derived from Ganoderma lucidum provided by the present invention can solve the problems of poor oral mucosal permeability of protein, easy degradation in the gastrointestinal tract, and low bioavailability.

[0008] To achieve the above object, the present invention adopts the following technical solution:

[0009] On the one hand, the present invention provides a protein orally soluble film derived from Ganoderma lucidum, which comprises the following raw materials, measured by mass percentage: 1-10% of microsporous Ganoderma lucidum immune protein, 25%-50% of film-forming material, 5%-20% of plasticizer, 5%-30% of mucosal adhesive, 1%-20% of penetration enhancer, 5%-20% of flavoring agent, 0.1%-1% of colorant and 0.5%-3% of acidity regulator; the microsporous Ganoderma lucidum immune protein is microsporous Ganoderma lucidum immune protein freeze-dried powder or microsporous Ganoderma lucidum immune protein composite nanoparticles.

[0010] Preferably, the mass ratio of the film-forming material to the mucosal adhesive is 1-3:1.

[0011] Preferably, the protein orally soluble film derived from Ganoderma lucidum comprises the following raw materials, calculated by mass percentage: 3% microspore Ganoderma lucidum immune protein, 38% film-forming material, 8% plasticizer, 20% mucosal adhesive, 15% penetration enhancer, 14.7% flavoring agent, 0.3% colorant and 1% acidity regulator.

[0012] The film-forming material is the skeleton component of the oral film and can form a film with a complete structure and rapid dissolution.

[0013] Preferably, the film-forming material used is selected from one or more of hydroxypropyl methylcellulose (HPMC), polyvinyl alcohol (PVA), hydroxypropyl cellulose (HPC), polyvinyl pyrrolidone (PVP), polyethylene oxide (PEO), hydroxyethyl cellulose (HEC), sodium alginate, and chitosan.

[0014] More preferably, the film-forming material used is selected from one or more of hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), polyvinyl pyrrolidone (PVP), hydroxyethyl cellulose (HEC), sodium alginate, and chitosan.

[0015] Further preferably, the film-forming materials used are hydroxypropyl methylcellulose, sodium alginate and chitosan. The plasticizer is used to improve the flexibility of the film to prevent the oral film from being easily broken during transportation or use.

[0016] Preferably, the plasticizer used is selected from one or more of glycerol, propylene glycol, polyethylene glycol 400 or 600, sorbitol, and glyceryl monostearate (GMS).

[0017] More preferably, the plasticizer used is selected from one or more of glycerol, polyethylene glycol 400 or 600, and glyceryl monostearate (GMS).

[0018] More preferably, the plasticizers used are glycerol and polyethylene glycol 400.

[0019] The mucoadhesive can enhance the adhesion between the oral membrane and the oral mucosa, thereby prolonging the drug residence time and improving the absorption efficiency.

[0020] Preferably, the mucoadhesive used is selected from one or more of polyvinyl pyrrolidone (PVP), sodium alginate, chitosan, sodium carboxymethyl cellulose (CMC-Na), carbomer (Carbopol), and gelatin.

[0021] More preferably, the mucosal adhesive used is selected from one or more of polyvinyl pyrrolidone (PVP), sodium carboxymethyl cellulose (CMC-Na), carbomer (Carbopol), and gelatin.

[0022] More preferably, the mucoadhesive agents used are polyvinyl pyrrolidone (PVP) and carbopol.

[0023] The penetration enhancer used is used to increase the permeability of GMI through the oral mucosa, thereby promoting drug absorption.

[0024] Preferably, the penetration enhancer is selected from one or more of sodium dodecyl sulfate (SDS), sodium deoxycholate, sodium glycocholate, menthol, Tween surfactants (such as Tween 20 or Tween 80), glycerol fatty acid esters, and sodium 8-(2-hydroxybenzamido) caprylate (SNAC).

[0025] More preferably, the penetration enhancer is selected from one or more of sodium dodecyl sulfate (SDS), Tween surfactants (such as Tween 20 or Tween 80), glycerol fatty acid esters, and sodium 8-(2-hydroxybenzamido) caprylate (SNAC).

[0026] More preferably, the penetration enhancer is Tween 80 and glycerol fatty acid ester.

[0027] The flavoring agent used is used to improve the mouthfeel of the oral film and increase the compliance of use.

[0028] Preferably, the flavoring agent is selected from one or more of xylitol, stevioside, sucralose, aspartame, mint flavor, and lemon flavor.

[0029] More preferably, the flavoring agent is selected from one or more of xylitol, stevioside, mint flavor, and lemon flavor.

[0030] More preferably, the flavoring agent is xylitol, steviol glycoside and mint essence.

[0031] Colorants are used to improve the appearance of the oral film.

[0032] Preferably, the colorant is selected from one or more of lemon yellow, carmine, sunset yellow, indigo and titanium dioxide.

[0033] More preferably, the colorant is selected from one or more of lemon yellow, carmine, and indigo.

[0034] More preferably, the colorant is lemon yellow.

[0035] The acidity regulator used is used to control the pH value of the oral film and optimize solubility and taste.

[0036] Preferably, the acidity regulator is selected from one or more of citric acid, sodium citrate, tartaric acid and lactic acid.

[0037] Further preferably, the acidity regulator is citric acid, lactic acid and tartaric acid.

[0038] In a second aspect, the present invention also provides a method for preparing microspore Ganoderma lucidum immune protein composite nanoparticles, comprising the following steps:

[0039] S1. Preparation of zein ethanol solution: Add zein to the ethanol solution and stir to mix;

[0040] S2. Preparation of GMI-zein binary nanoparticles (GMI-Z): dissolving GMI in deionized water or buffer to form a GMI solution, then adding the GMI solution dropwise to the zein ethanol solution obtained in step S1, stirring and mixing to form a GMI-zein binary nanoparticle solution;

[0041] S3. Preparation of GMI-zein-sodium alginate ternary nanoparticles (GMI-Z-SA): dissolving sodium alginate in deionized water to form a sodium alginate solution; dropping the GMI-zein binary nanoparticle solution obtained in step S2 into the sodium alginate solution, stirring and mixing to form a GMI-zein-sodium alginate ternary nanoparticle solution;

[0042] S4. Removal of ethanol and particle stabilization: The GMI-zein-sodium alginate ternary nano-solution obtained in step S3 is rotary evaporated until the ethanol is completely removed to obtain a nanoparticle suspension; the nanoparticle suspension is allowed to stand, centrifuged, and repeated 2-3 times; and then freeze-dried to obtain a GMI composite nanoparticle solid powder.

[0043] Preferably, the concentration of ethanol in step S1 is 70%-80%, preferably 80%;

[0044] Preferably, the concentration of zein in step S1 is 4%-6%, preferably 5%.

[0045] Preferably, the volume ratio of the GMI solution to the zein ethanol solution in step S2 is 1:2-4; preferably 1:3.

[0046] Preferably, the stirring in step S2 is carried out by rapid oscillation in a vortex machine or magnetic stirring, and the stirring rate of the magnetic stirring is 3000-5000 r / min.

[0047] Preferably, the volume ratio of the GMI-zein binary nano solution to the sodium alginate solution in step S3 is 1:3-5, preferably 1:4.

[0048] Preferably, the stirring rate in step S3 is 8000-12000 r / min.

[0049] Preferably, the temperature of the rotary evaporation in step S4 is 40° C. and the pressure is -0.1 MPa; the standing time is 30-60 min; the centrifugal speed is 3000-5000 rpm, and the centrifugal time is 10-15 min.

[0050] In a third aspect, the present invention further provides a method for preparing a protein oral dissolving film derived from Ganoderma lucidum, comprising the following steps:

[0051] (1) Dissolve glycerol and glycerol fatty acid ester in a water bath and cool.

[0052] (2) Weighing a formulated amount of GMI lyophilized powder or GMI composite nanoparticle solid powder and dissolving it in purified water; then adding a plasticizer, a mucoadhesive agent, a penetration enhancer, a flavoring agent, a colorant and an acidity regulator, stirring and mixing uniformly to obtain a mixed aqueous solution;

[0053] (3) adding a film-forming material to the mixed aqueous solution obtained in step (2), stirring and dispersing the film-forming material to form a uniform slurry;

[0054] (4) placing the slurry obtained in step (3) in a vacuum degassing device to remove bubbles, thereby obtaining a degassing slurry;

[0055] (5) The defoamed slurry obtained in step (4) is coated on a substrate, heated and dried to form a thin film, and then the film is peeled off from the substrate for slitting and packaging.

[0056] Preferably, the substrate in step (5) comprises one of polyethylene terephthalate (PET) and polyethylene (PE).

[0057] Preferably, the drying temperature in step (5) is 40-60°C.

[0058] Preferably, the thickness of the film in step (5) is 50-200 μm.

[0059] Finally, the present invention also provides the application of the orodispersible protein film derived from Ganoderma lucidum in food, health products or medicines.

[0060] Compared with the prior art, the present invention has the following beneficial effects:

[0061] (1) The present invention embeds the GMI protein in composite nanoparticles, which can effectively improve its stability, reduce degradation and enhance bioavailability.

[0062] (2) The preparation process of the present invention is simple and the conditions are mild, which effectively protects the biological activity of the protein and can realize industrial mass production, thereby reducing production costs.

[0063] (3) The present invention optimizes the types and ratios of the formula components, thereby ensuring that the orodispersible film not only has excellent appearance quality, but also has good folding resistance and short disintegration time, further improving the user experience of the product.

[0064] (4) The present invention significantly improves the bioavailability of GMI protein through two pathways: direct absorption through the oral mucosa and absorption through the gastrointestinal tract. DETAILED DESCRIPTION

[0065] The technical scheme of the present invention is clearly and completely described below with specific implementation methods. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without creative work are within the scope of protection of the present invention. The materials, reagents, etc. used, if not otherwise specified, are reagents and materials available from commercial sources.

[0066] Basic Example 1 Preparation of GMI composite nanoparticles (GMI-CNPs)

[0067] The GMI solution with a concentration of 5 mg / mL was slowly added dropwise to a 5% (m:v) zein ethanol solution (80% ethanol, v:v) at a volume ratio of 1:3, and rapidly oscillated with a vortex instrument to form a GMI-Z binary mixed solution. Subsequently, the mixed solution was added dropwise to a 0.05% (m:v) sodium alginate aqueous solution at a final volume ratio of 1:4, and stirred at a speed of 10000 r / min while adding dropwise until GMI-zein-sodium alginate composite nanoparticles were formed. Finally, the GMI-zein-sodium alginate composite nanoparticle solution was rotary evaporated to remove ethanol at 40°C and -0.1MPa, and lyophilized to obtain GMI composite nanoparticles for subsequent use.

[0068] The formulations of Examples 1-6 are shown in Table 1 below:

[0069] Table 1

[0070]

[0071]

[0072] The formulations of Examples 7-11 are shown in Table 2 below:

[0073] Table 2

[0074]

[0075] The formulas of Comparative Examples 1-4 are shown in Table 3 below:

[0076] Table 3

[0077]

[0078]

[0079] The formulas of Comparative Examples 5-8 are shown in Table 4 below:

[0080] Table 4

[0081]

[0082] The formulations of Examples 1-11 and Comparative Examples 1-8 were prepared into orally soluble protein films derived from Ganoderma lucidum according to the following method, specifically:

[0083] (1) Dissolve glycerol and glycerol fatty acid ester in a 50°C water bath to form a uniform solution, and cool to 40°C for later use.

[0084] (2) Weighing a formulated amount of GMI lyophilized powder or GMI composite nanoparticle solid powder and dissolving it in purified water; then adding polyethylene glycol 400, polyvinyl pyrrolidone, carbomer, glycerol fatty acid ester, Tween 80, citric acid, tartaric acid, lactic acid, xylitol, steviol glycoside, and tartrazine, stirring and mixing uniformly to obtain a mixed aqueous solution;

[0085] (3) adding sodium alginate, chitosan, hydroxypropyl methylcellulose and hydroxypropyl cellulose to the mixed aqueous solution obtained in step (2), stirring and dispersing the mixture to form a uniform slurry;

[0086] (4) placing the slurry obtained in step (3) in a vacuum degassing device for degassing for 30 minutes to obtain a degassing slurry;

[0087] (5) coating the defoamed slurry obtained in step (4) on a substrate at a coating speed of 20-40 mm / s and a drying temperature of 40° C., controlling the film thickness to be about 1 mm, heating and drying to form a thin film, then peeling the film from the substrate, cutting it into tablets of the same specification, and packaging them individually.

[0088] Effect Experiment

[0089] 1. Investigation of adhesion performance and adhesion time

[0090] The orodispersible films obtained in Examples 1, 2, 6 and 7-11 and Comparative Examples 1, 2 and 5-8 were selected, and three volunteers aged 25-28 were selected to stick the films on the buccal mucosa moistened with saliva, and the adhesion performance was evaluated and the adhesion time was recorded. Among them, the adhesion time was divided into three levels, adhesion of more than 30 minutes was good, adhesion of 10-30 minutes was good, and adhesion of less than 10 minutes was poor. The test results are shown in Tables 5 and 7.

[0091] Table 5

[0092]

[0093] The results in Table 5 show that the ratio of film-forming material to mucoadhesive has a better adhesion effect in the range of 1:1-3:1. When the ratio exceeds this range, the adhesion time will decrease. In addition, if the proportion of mucoadhesive is too large, the slurry will gel, the viscosity will be too high, the fluidity will become poor, and it will be difficult to apply the film.

[0094] 2. Performance Testing

[0095] (1) Melting time detection

[0096] The orally dissolving films prepared in each embodiment and comparative example were placed in a culture dish containing 20 mL of purified water at a temperature of 37±0.5°C, and the time for the sample to completely dissolve was recorded, which was the disintegration time. Among them, a dissolution time of ≤50s means fast dissolution; a dissolution time of 50s-120s means moderate dissolution; and a dissolution time of >120s means slow dissolution.

[0097] (2) Tensile strength test

[0098] Testing method: Tensile strength is also called strength limit, which refers to the maximum force used to break the oral film. Use an intelligent electronic tensile testing machine for testing.

[0099] Tensile strength calculation formula: Tensile strength = applied stress / cross-sectional area.

[0100] Among them, films with low tensile strength are relatively brittle.

[0101] (3) Film thickness detection

[0102] Use CHY-CU film thickness gauge to measure, take a sample with a width of 100mm, without wrinkles or other defects, place it on the test bench, and start measuring. The instrument automatically calculates the sample results.

[0103] (4) Physical properties test of membrane

[0104] Five volunteers were selected to score the strength and flexibility of the membrane, which were divided into three levels: good (≥90 points), better (85-95 points), worse (70-85 points), and poor (≤70 points).

[0105] The performance test results of samples with different prescriptions are shown in Tables 6 and 7.

[0106] Table 6

[0107]

[0108] The results in Table 6 show that the orodissolving films prepared in Examples 1-6 have good flexibility, mechanical properties and disintegration. Comparative Example 3 has poor film-forming properties due to the low content of sodium alginate, a film-forming material, and the oral film is prone to fragmentation. Comparative Example 4 has low strength after film formation due to the low content of plasticizer glycerol, and is easily damaged during use. Comparative Example 3 has high content of plasticizer glycerol, which affects the uniformity of orodissolving film disintegration.

[0109] Table 7

[0110]

[0111]

[0112] It can be seen from the data in Table 7 that the orodispersible films prepared in Examples 7-11 have good flexibility, mechanical properties and solubility. In Comparative Example 5, the proportion of film-forming materials is low, the film is fragile and easy to break, and the excessive amount of carbomer improves the mucosal adhesion, but causes the film surface to be too sticky and affects the dissolution rate of the orodispersible film; in Comparative Example 6, the proportion of sodium alginate is too high, resulting in increased hardness and insufficient toughness of the film; in Comparative Example 7, the proportion of PVP is too high, although the adhesion is improved, the mechanical strength of the film is affected; in Comparative Example 8, the proportion of chitosan is too high, which affects the transparency and solubility of the film.

[0113] In summary, the orodispersible film prepared in the embodiment of the present invention has a simple preparation process, good flexibility, mechanical properties and disintegration, and has broad application prospects.

Claims

1. A protein orally dissolving film derived from Ganoderma lucidum, characterized in that: Calculated by mass percentage, the raw materials include: 1-10% microspore Ganoderma lucidum immune protein, 25%-50% film-forming material, 5%-20% plasticizer, 5%-30% mucosal adhesive, 1%-20% penetration enhancer, 8%-20% flavoring agent, 0.1%-1% colorant and 0.5%-3% acidity regulator.

2. The protein orally dissolving film derived from Ganoderma lucidum according to claim 1, characterized in that: Calculated by mass percentage, the raw materials include: 3% microspore Ganoderma lucidum immune protein, 38% film-forming material, 8% plasticizer, 20% mucosal adhesive, 15% penetration enhancer, 14.7% flavoring agent, 0.3% colorant and 1% acidity regulator.

3. The orodispersible protein film derived from Ganoderma lucidum according to claim 1, characterized in that: The microsporous ganoderma lucidum immune protein is microsporous ganoderma lucidum immune protein freeze-dried powder or microsporous ganoderma lucidum immune protein composite nanoparticles.

4. The orodispersible protein film derived from Ganoderma lucidum according to claim 3, characterized in that: The microspore Ganoderma lucidum immune protein composite nanoparticles are prepared by the following method: S1. Preparation of zein ethanol solution: adding zein at a concentration of 4% to 6% to an ethanol solution at a concentration of 70% to 80%, stirring and mixing to obtain a zein ethanol solution; S2. Preparation of Ganoderma lucidum protein-zein binary nanoparticles: dissolving the Ganoderma lucidum protein in deionized water or buffer to form a Ganoderma lucidum protein solution, and then mixing the Ganoderma lucidum protein solution and the zein ethanol solution obtained in step S1 in a volume ratio of 1:2-4 to form a Ganoderma lucidum protein-zein binary nanoparticle solution; S3. Preparation of Ganoderma lucidum protein-zein-sodium alginate ternary nanoparticles: dissolving sodium alginate in deionized water to form a sodium alginate solution; mixing the Ganoderma lucidum protein-zein binary nanoparticle solution and the sodium alginate solution obtained in step S2 in a volume ratio of 1:3-5 to form a Ganoderma lucidum protein-zein-sodium alginate ternary nanoparticle solution; S4. Removal of ethanol and particle stabilization: The protein-zein-sodium alginate ternary nano-solution derived from Ganoderma lucidum obtained in step S3 is subjected to rotary evaporation at a temperature of 35-40°C and a pressure of -0.1 MPa until the ethanol is completely removed to obtain a nanoparticle suspension; the nanoparticle suspension is allowed to stand for 30-60 minutes, centrifuged at 3000-5000 rpm for 10-15 minutes, and repeated 2-3 times; and then freeze-dried to obtain a solid powder of protein composite nanoparticles derived from Ganoderma lucidum.

5. The orodispersible protein film derived from Ganoderma lucidum according to claim 1, characterized in that: The film-forming material is selected from one or more of hydroxypropyl methylcellulose, polyvinyl alcohol, hydroxypropyl cellulose, polyvinyl pyrrolidone, polyethylene oxide, hydroxyethyl cellulose, sodium alginate and chitosan.

6. The orodispersible protein film derived from Ganoderma lucidum according to claim 1, characterized in that: The plasticizer is selected from one or more of glycerol, propylene glycol, polyethylene glycol, sorbitol, and glyceryl monostearate.

7. The orodispersible protein film derived from Ganoderma lucidum according to claim 1, characterized in that: The mucosal adhesive is selected from one or more of polyvinyl pyrrolidone, sodium alginate, chitosan, sodium carboxymethyl cellulose, carbomer, and gelatin.

8. The orodispersible protein film derived from Ganoderma lucidum according to claim 1, characterized in that: The penetration enhancer is selected from one or more of sodium dodecyl sulfate, sodium deoxycholate, sodium glycocholate, menthol, Tween 20, Tween 80, glycerol fatty acid ester, and sodium 8-(2-hydroxybenzamido) caprylate.

9. The orodispersible protein film derived from Ganoderma lucidum according to claim 1, characterized in that: The flavoring agent is selected from one or more of xylitol, stevioside, sucralose, aspartame, mint flavor, lemon flavor; the coloring agent is selected from one or more of tartrazine, carmine, sunset yellow, indigo, titanium dioxide; the acidity regulator is selected from one or more of citric acid, sodium citrate, tartaric acid, lactic acid.

10. The method for preparing the orodispersible protein film derived from Ganoderma lucidum according to any one of claims 1 to 9, characterized in that: The steps include: (1) weighing a formulated amount of microspore Ganoderma lucidum immune protein and dissolving it in purified water; then adding a plasticizer, a flavoring agent, a mucoadhesive agent, a penetration enhancer, a flavoring agent, a colorant and an acidity regulator, stirring and mixing uniformly to obtain a mixed aqueous solution; (2) adding a film-forming agent to the mixed aqueous solution obtained in step (1), stirring and dispersing the mixture to form a uniform slurry; (3) degassing the slurry obtained in step (2) to obtain a degassing slurry; (4) coating the defoamed slurry obtained in step (3) on a substrate, heating and drying to form a thin film, and then peeling the film from the substrate for slitting and packaging.

11. Use of the protein orally dissolving film derived from Ganoderma lucidum according to any one of claims 1 to 9 or the protein orally dissolving film derived from Ganoderma lucidum prepared by the preparation method according to claim 10 in preparing food, health products or medicines.

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