Liver-protecting polypeptide preparation and preparation method thereof

Abstract

The invention relates to the technical field of medicines, in particular to a liver-protecting polypeptide preparation and a preparation method thereof. The pharmaceutical composition comprises a polypeptide with the following amino acid sequence: Gly-Ala-Leu-Pro-Ser-Asp-Gly-Glu-Arg-Lys-Gln-Phe-Ser-Gly-Arg-Leu-Ala-Gly-Lys-Ser (SEQ ID NO: 1), and a pharmaceutically acceptable carrier, and the carrier comprises the following auxiliary materials: hydroxypropyl methyl cellulose (HPMC), polyvinylpyrrolidone (PVP), mannitol, microcrystalline cellulose, lactose and polyethylene glycol (PEG). The invention provides a novel liver-protecting polypeptide preparation and a preparation method thereof, the preparation comprises a polypeptide with a specific amino acid sequence, the polypeptide has a definite liver-protecting effect, and high purity and stability of the medicine are ensured through the optimized preparation method.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a liver-protecting polypeptide preparation and a preparation method thereof. Background Art

[0002] Liver disease is a major public health problem worldwide, including hepatitis, fatty liver, cirrhosis and liver cancer. Traditional therapeutic drugs such as antiviral drugs, antioxidants and anti-inflammatory drugs can relieve symptoms to a certain extent, but the effect is limited and there are side effects. In recent years, peptide drugs have gradually become a new direction for the treatment of liver disease due to their high specificity and low toxicity.

[0003] Peptides are biological macromolecules composed of amino acids connected by peptide bonds, and have diverse biological activities. Currently, a variety of peptide drugs have been used in clinical practice, such as GLP-1 receptor agonists for the treatment of type 2 diabetes, and insulin. However, there are relatively few peptide drugs specifically used for liver protection, and the efficacy and safety of existing drugs still need to be improved.

[0004] In the field of liver disease treatment, although peptide drugs have shown their unique advantages, there are still some technical defects in the peptide drug preparations for liver disease on the market. First, many peptide drugs have poor stability and are easily enzymatically hydrolyzed in the body, resulting in unstable efficacy. Secondly, the molecular weight of peptide drugs is large and it is difficult for them to pass through the cell membrane, which affects their distribution and absorption in the liver. In addition, the preparation cost of peptide drugs is high, and impurities are easily generated during the preparation process, affecting the purity and safety of the drugs. Finally, most of the existing peptide drug preparations are injectable, which brings inconvenience to patients, especially those who need long-term treatment. Summary of the invention

[0005] The present invention provides a new hepatoprotective polypeptide preparation and a preparation method thereof. The preparation comprises a polypeptide with a specific amino acid sequence, which has a clear hepatoprotective effect, and the high purity and stability of the drug are ensured by an optimized preparation method.

[0006] The technical solution adopted by the present invention is: a liver-protecting polypeptide preparation, comprising a polypeptide having the following amino acid sequence: Gly-Ala-Leu-Pro-Ser-Asp-Gly-Glu-Arg-Lys-Gln-Phe-Ser-Gly-Arg-Leu-Ala-Gly-Lys-Ser (SEQ ID NO: 1), and a pharmaceutically acceptable carrier, wherein the molecular weight of the polypeptide is 1500-2000 Daltons and the purity is not less than 95%.

[0007] As a further improvement of the present invention, the mass percentage of the polypeptide in the preparation is 1-10%.

[0008] As a further improvement of the present invention, the carrier comprises the following excipients: hydroxypropyl methylcellulose (HPMC), polyvinyl pyrrolidone (PVP), mannitol, microcrystalline cellulose, lactose and polyethylene glycol (PEG).

[0009] As a further improvement of the present invention, the mass percentages of the excipients in the preparation are: hydroxypropyl methylcellulose (HPMC) accounts for 15%-25%, polyvinyl pyrrolidone (PVP) accounts for 5%-15%, mannitol accounts for 25%-35%, microcrystalline cellulose accounts for 10%-20%, lactose accounts for 5%-10%, polyethylene glycol (PEG) accounts for 1%-5%, and the balance is purified water.

[0010] As a further improvement of the present invention, the dosage form of the preparation is an oral liquid.

[0011] A method for preparing a liver-protecting polypeptide preparation comprises the following steps:

[0012] Step 1, peptide synthesis: using solid phase synthesis method, using Wang resin as a carrier, sequentially coupling amino acids to synthesize a peptide with a specified sequence, and performing purification after each coupling reaction;

[0013] Step 2, polypeptide purification: high performance liquid chromatography (HPLC) is used to purify the synthesized polypeptide to ensure that the purity of the polypeptide reaches more than 95%;

[0014] Step 3, preparation of preparation: mixing the purified polypeptide with the excipients according to the above mass percentage to obtain a uniform mixture;

[0015] Step 4: Forming the dosage form: Add purified water to the mixture, stir evenly, filter, and then fill it through oral liquid preparation equipment to obtain the final oral liquid preparation.

[0016] As a further improvement of the present invention, the usage amount of the Wang resin is 10%-20% of the total mass of the polypeptide synthesis.

[0017] As a further improvement of the present invention, the high performance liquid chromatography (HPLC) adopts a C18 reverse phase chromatographic column, and uses acetonitrile and water as mobile phases for gradient elution, with a flow rate of 1.0 to 1.5 mL / min and a detection wavelength of 220 nm.

[0018] As a further improvement of the present invention, the mixing process is carried out by a high-speed mixer, and the mixing time is 20-30 minutes; the filtering process uses a 0.22 micron microporous filter membrane to remove particulate impurities therein.

[0019] Beneficial effects of the present invention: (1) The polypeptide preparation of the present invention has a clear liver-protecting effect. Through the specific amino acid sequence design, it can effectively protect liver cells, reduce liver cell damage, and promote liver function recovery, thereby significantly improving the treatment effect and quality of life of patients with liver disease.

[0020] (2) The present invention ensures the high purity and stability of the polypeptide preparation by optimizing the preparation method, and adopts solid phase synthesis and high performance liquid chromatography to synthesize and purify the polypeptide, effectively removing impurities and improving the purity of the drug. At the same time, through reasonable selection and proportion of excipients, the stability of the drug is enhanced, the shelf life of the drug is extended, and the drug is not easily deteriorated or degraded during storage and use.

[0021] (3) The polypeptide preparation of the present invention is in the form of an oral liquid, which is convenient for patients to take and improves patient compliance. The oral liquid avoids the pain and inconvenience caused by injection and is particularly suitable for patients who require long-term treatment. In addition, the oral liquid can also be dosed according to the patient's specific condition and medication needs, thereby improving the flexibility and personalization of medication. DETAILED DESCRIPTION

[0022] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present application more clearly understood, the present application is further described in detail below in conjunction with the embodiments. It should be understood that the embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0023] The present invention provides a liver-protecting polypeptide preparation, characterized in that it comprises a polypeptide having the following amino acid sequence: Gly-Ala-Leu-Pro-Ser-Asp-Gly-Glu-Arg-Lys-Gln-Phe-Ser-Gly-Arg-Leu-Ala-Gly-Lys-Ser (SEQ ID NO: 1), and a pharmaceutically acceptable carrier, the molecular weight of the polypeptide is 1500-2000 Daltons, the purity is not less than 95%, and the mass percentage of the polypeptide in the preparation is 1-10%.

[0024] The carrier of the present invention comprises the following excipients: hydroxypropyl methylcellulose (HPMC), polyvinyl pyrrolidone (PVP), mannitol, microcrystalline cellulose, lactose and polyethylene glycol (PEG); the mass percentages of the excipients in the preparation are: hydroxypropyl methylcellulose (HPMC) accounts for 15%-25%, polyvinyl pyrrolidone (PVP) accounts for 5%-15%, mannitol accounts for 25%-35%, microcrystalline cellulose accounts for 10%-20%, lactose accounts for 5%-10%, polyethylene glycol (PEG) accounts for 1%-5%, and the balance is purified water.

[0025] The dosage form of the preparation of the present invention is oral liquid.

[0026] A method for preparing a liver-protecting polypeptide preparation comprises the following steps:

[0027] Step 1, peptide synthesis: using solid phase synthesis method, using Wang resin as a carrier, sequentially coupling amino acids to synthesize a peptide with a specified sequence, and performing purification after each coupling reaction. The amount of Wang resin used is 10%-20% of the total mass of the peptide synthesis;

[0028] Step 2, polypeptide purification: the synthesized polypeptide is purified by high performance liquid chromatography (HPLC) to ensure that the purity of the polypeptide reaches more than 95%, the high performance liquid chromatography (HPLC) adopts a C18 reverse phase chromatography column, and acetonitrile and water are used as mobile phases for gradient elution, the flow rate is 1.0 to 1.5 mL / min, and the detection wavelength is 220 nm;

[0029] Step 3, preparation of preparation: the purified polypeptide is mixed with the excipients according to the above mass percentage, the mixing process is carried out by a high-speed mixer, the mixing time is 20-30 minutes, and a uniform mixture is obtained;

[0030] Step 4: Forming the dosage form: Add purified water to the mixture, stir evenly and filter. The filtration process uses a 0.22-micron microporous filter membrane to remove particulate impurities. The mixture is then filled through an oral liquid preparation device to obtain a final oral liquid preparation.

[0031] Embodiment 1:

[0032] This embodiment provides a method for preparing a liver-protecting polypeptide preparation, and the specific steps are as follows.

[0033] Step 1: Peptide synthesis

[0034] The solid phase synthesis method is adopted, Wang resin is used as a carrier, and amino acids are coupled in sequence according to the specified amino acid sequence (Gly-Ala-Leu-Pro-Ser-Asp-Gly-Glu-Arg-Lys-Gln-Phe-Ser-Gly-Arg-Leu-Ala-Gly-Lys-Ser, SEQ ID NO: 1); after each step of coupling reaction, purification treatment is performed to remove unreacted amino acids and by-products; the amount of Wang resin used is controlled at about 15% of the total mass of the polypeptide synthesis to ensure the effectiveness and yield of the reaction.

[0035] Step 2: Peptide purification

[0036] The synthesized peptide was purified by high performance liquid chromatography (HPLC); a C18 reverse phase column was used, acetonitrile and water were used as mobile phases for gradient elution, and the flow rate was set at 1.2 mL / min; the detection wavelength was 220 nm, ensuring that the purity of the peptide reached more than 95%.

[0037] Step 3: Preparation of formulation

[0038] The purified polypeptide was mixed with the excipients according to the following mass percentages: 20% hydroxypropyl methylcellulose (HPMC), 10% polyvinyl pyrrolidone (PVP), 30% mannitol, 15% microcrystalline cellulose, 7% lactose, 3% polyethylene glycol (PEG), and the balance was purified water; the mixing process was carried out using a high-speed mixer for 25 minutes to ensure a uniform drug mixture.

[0039] Step 4: Dosage Forming

[0040] The drug mixture is added into purified water and stirred evenly; filtered using a 0.22 micron microporous filter membrane to remove particulate impurities therein; and filled through oral liquid preparation equipment to obtain a final oral liquid preparation.

[0041] The oral liquid preparation prepared by the above steps has a polypeptide mass percentage of 5% and a purity of not less than 95%, and has a clear liver-protecting effect.

[0042] Embodiment 2:

[0043] This embodiment provides a method for preparing a liver-protecting polypeptide preparation, and the specific steps are as follows.

[0044] Step 1: Peptide synthesis

[0045] The solid phase synthesis method is adopted, Wang resin is used as a carrier, and amino acids are coupled in sequence according to the specified amino acid sequence (Gly-Ala-Leu-Pro-Ser-Asp-Gly-Glu-Arg-Lys-Gln-Phe-Ser-Gly-Arg-Leu-Ala-Gly-Lys-Ser, SEQ ID NO: 1); after each step of coupling reaction, purification treatment is performed to remove unreacted amino acids and by-products; the amount of Wang resin used is controlled at about 18% of the total mass of polypeptide synthesis to ensure the effectiveness and yield of the reaction.

[0046] Step 2: Peptide purification

[0047] The synthesized peptide was purified by high performance liquid chromatography (HPLC); a C18 reverse phase column was used, acetonitrile and water were used as mobile phases for gradient elution, and the flow rate was set at 1.3 mL / min; the detection wavelength was 220 nm to ensure that the purity of the peptide reached more than 95%.

[0048] Step 3: Preparation of formulation

[0049] The purified polypeptide was mixed with the excipients according to the following mass percentages: 18% hydroxypropyl methylcellulose (HPMC), 8% polyvinyl pyrrolidone (PVP), 28% mannitol, 18% microcrystalline cellulose, 8% lactose, 2% polyethylene glycol (PEG), and the balance was purified water; the mixing process was carried out using a high-speed mixer for 28 minutes to ensure a uniform drug mixture.

[0050] Step 4: Dosage Forming

[0051] The drug mixture is added into purified water and stirred evenly; filtered using a 0.22 micron microporous filter membrane to remove particulate impurities therein; and filled through oral liquid preparation equipment to obtain a final oral liquid preparation.

[0052] The oral liquid preparation prepared by the above steps has a polypeptide mass percentage of 4% and a purity of not less than 95%, and has a clear liver-protecting effect.

[0053] Embodiment 3:

[0054] This embodiment provides a method for preparing a liver-protecting polypeptide preparation, and the specific steps are as follows.

[0055] Step 1: Peptide synthesis

[0056] The solid phase synthesis method is adopted, Wang resin is used as a carrier, and amino acids are coupled in sequence according to the specified amino acid sequence (Gly-Ala-Leu-Pro-Ser-Asp-Gly-Glu-Arg-Lys-Gln-Phe-Ser-Gly-Arg-Leu-Ala-Gly-Lys-Ser, SEQ ID NO: 1); after each step of coupling reaction, purification treatment is carried out to remove unreacted amino acids and by-products; the amount of Wang resin used is controlled at about 16% of the total mass of polypeptide synthesis to ensure the effectiveness and yield of the reaction.

[0057] Step 2: Peptide purification

[0058] The synthesized peptide was purified by high performance liquid chromatography (HPLC); a C18 reverse phase column was used, acetonitrile and water were used as mobile phases for gradient elution, and the flow rate was set at 1.1 mL / min; the detection wavelength was 220 nm to ensure that the purity of the peptide reached more than 95%.

[0059] Step 3: Preparation of formulation

[0060] The purified polypeptide was mixed with the excipients according to the following mass percentages: 24% hydroxypropyl methylcellulose (HPMC), 12% polyvinyl pyrrolidone (PVP), 32% mannitol, 17% microcrystalline cellulose, 6% lactose, 4% polyethylene glycol (PEG), and the balance was purified water; the mixing process was carried out using a high-speed mixer for 22 minutes to ensure a uniform drug mixture.

[0061] Step 4: Dosage Forming

[0062] The drug mixture is added into purified water and stirred evenly; filtered using a 0.22 micron microporous filter membrane to remove particulate impurities therein; and filled through oral liquid preparation equipment to obtain a final oral liquid preparation.

[0063] The oral liquid preparation prepared by the above steps has a polypeptide mass percentage of 6% and a purity of not less than 95%, and has a clear liver-protecting effect.

[0064] Embodiment 4:

[0065] This embodiment provides a method for preparing a liver-protecting polypeptide preparation, and the specific steps are as follows.

[0066] The solid phase synthesis method is adopted, Wang resin is used as a carrier, and amino acids are coupled in sequence according to the specified amino acid sequence (Gly-Ala-Leu-Pro-Ser-Asp-Gly-Glu-Arg-Lys-Gln-Phe-Ser-Gly-Arg-Leu-Ala-Gly-Lys-Ser, SEQ ID NO: 1); after each step of coupling reaction, purification treatment is performed to remove unreacted amino acids and by-products; the amount of Wang resin used is controlled at about 15% of the total mass of the polypeptide synthesis to ensure the effectiveness and yield of the reaction.

[0067] Step 2: Peptide purification

[0068] The synthesized peptide was purified by high performance liquid chromatography (HPLC); a C18 reverse phase column was used, acetonitrile and water were used as mobile phases for gradient elution, and the flow rate was set at 1.4 mL / min; the detection wavelength was 220 nm to ensure that the purity of the peptide reached more than 95%.

[0069] Step 3: Preparation of formulation

[0070] The purified polypeptide was mixed with the excipients according to the following mass percentages: 22% hydroxypropyl methylcellulose (HPMC), 11% polyvinyl pyrrolidone (PVP), 26% mannitol, 13% microcrystalline cellulose, 6% lactose, 2% polyethylene glycol (PEG), and the balance was purified water; the mixing process was carried out using a high-speed mixer for 26 minutes to ensure a uniform drug mixture.

[0071] Step 4: Dosage Forming

[0072] The drug mixture is added into purified water and stirred evenly; filtered using a 0.22 micron microporous filter membrane to remove particulate impurities therein; and filled through oral liquid preparation equipment to obtain a final oral liquid preparation.

[0073] The oral liquid preparation prepared by the above steps has a polypeptide mass percentage of 5% and a purity of not less than 95%, and has a clear liver-protecting effect.

[0074] Embodiment 5:

[0075] This embodiment provides a method for preparing a liver-protecting polypeptide preparation, and the specific steps are as follows.

[0076] The solid phase synthesis method is adopted, Wang resin is used as a carrier, and amino acids are coupled in sequence according to the specified amino acid sequence (Gly-Ala-Leu-Pro-Ser-Asp-Gly-Glu-Arg-Lys-Gln-Phe-Ser-Gly-Arg-Leu-Ala-Gly-Lys-Ser, SEQ ID NO: 1); after each step of coupling reaction, purification treatment is performed to remove unreacted amino acids and by-products; the amount of Wang resin used is controlled at about 15% of the total mass of the polypeptide synthesis to ensure the effectiveness and yield of the reaction.

[0077] Step 2: Peptide purification

[0078] The synthesized peptide was purified by high performance liquid chromatography (HPLC); a C18 reverse phase column was used, acetonitrile and water were used as mobile phases for gradient elution, and the flow rate was set at 1.2 mL / min; the detection wavelength was 220 nm, ensuring that the purity of the peptide reached more than 95%.

[0079] Step 3: Preparation of formulation

[0080] The purified polypeptide was mixed with the excipients according to the following mass percentages: 16% hydroxypropyl methylcellulose (HPMC), 9% polyvinyl pyrrolidone (PVP), 28% mannitol, 12% microcrystalline cellulose, 8% lactose, 3% polyethylene glycol (PEG), and the balance was purified water; the mixing process was carried out using a high-speed mixer for 25 minutes to ensure a uniform drug mixture.

[0081] Step 4: Dosage Forming

[0082] The drug mixture is added into purified water and stirred evenly; filtered using a 0.22 micron microporous filter membrane to remove particulate impurities therein; and filled through oral liquid preparation equipment to obtain a final oral liquid preparation.

[0083] The oral liquid preparation prepared by the above steps has a polypeptide mass percentage of 6% and a purity of not less than 95%, and has a clear liver-protecting effect.

[0084] Performance Testing

[0085] Oral solution stability test

[0086] Method: The prepared oral solution was tested for stability under different conditions (such as temperature, light, etc.).

[0087] Temperature: 4℃, 25℃, 10℃;

[0088] Light: Avoid light and light.

[0089] Results: The oral solution was stable at room temperature (25℃) and protected from light, without obvious precipitation or degradation.

[0090] (II) In vitro hepatocyte protection experiment

[0091] Methods: In vitro experiments were performed using human hepatocytes (such as HepG2 cells) to evaluate the protective effect of polypeptide preparations on hepatocytes.

[0092] The specific experimental steps are as follows:

[0093] (1) HepG2 cells were seeded in 96-well plates and cultured to 80% confluence.

[0094] (2) Use D-galactosamine (D-GalN) to induce hepatocyte damage.

[0095] (3) Add different concentrations of polypeptide preparations and continue culturing for 24 hours.

[0096] (4) The MTT method was used to detect cell activity, and the LDH release assay was used to detect the degree of cell damage.

[0097] The specific data is as follows:

[0098]

[0099]

[0100] Results: The peptide preparation had a significant protective effect on hepatocytes, significantly increased cell activity, and reduced LDH release.

[0101] (III) In vivo liver protection experiment

[0102] Methods: In vivo experiments were performed using a mouse model to evaluate the protective effect of peptide preparations on the liver.

[0103] The specific experimental steps are as follows:

[0104] (1) Establish a mouse model of acute liver injury (CCl4 induced).

[0105] (2) Administering peptide preparations and water control for a certain period of time.

[0106] (3) Detect the ALT and AST levels in mouse serum to assess the degree of liver damage.

[0107] (4) Histological examination (H&E staining) was used to evaluate liver tissue damage.

[0108] The specific data is as follows:

[0109]

[0110] Results: The ALT and AST levels of mice in the polypeptide preparation group were significantly lower than those in the control group, and the liver tissue damage was alleviated.

[0111] (IV) Pharmacokinetic studies

[0112] Methods: Pharmacokinetic studies were performed using mice to evaluate the absorption, distribution, metabolism, and excretion (ADME) of the peptide formulations.

[0113] The specific experimental steps are as follows:

[0114] (1) The peptide preparation was orally administered to mice.

[0115] (2) Blood was collected at different time points, and the concentration of peptides in plasma was detected using HPLC-MS / MS.

[0116] (3) Calculation of peptide pharmacokinetic parameters (Cmax, Tmax, AUC, etc.)

[0117] The specific data is as follows:

[0118] Time point (h) Peptide concentration in plasma (ng / mL) 0.5 25.3 1.0 50.4 2.0 75.6 4.0 60.2 8.0 30.1 12.0 15.0 24.0 7.5

[0119] Results: The peptide preparation showed good absorption and distribution characteristics in mice, with Cmax of 75.6 ng / mL, Tmax of 2 hours, and AUC of 450 ng·h / mL.

[0120] In summary, a hepatoprotective polypeptide preparation and a preparation method thereof of the present invention have significant pharmacokinetic properties, can be effectively absorbed by an organism and maintain a certain concentration in the body, thereby exerting its hepatoprotective therapeutic effect. In addition, the preparation of the present invention performed well in the stability test, indicating that it has good storage stability at room temperature and under light-proof conditions, and is suitable for long-term storage and use. Through in vitro hepatocyte protection experiments and in vivo liver protection experiments, the protective effect of the polypeptide preparation on hepatocytes and the improvement effect on liver damage in animal models were further verified. In summary, the present invention provides an effective and stable hepatoprotective polypeptide preparation and a preparation method thereof, which has good application prospects.

[0121] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features thereof may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A hepatoprotective polypeptide preparation, characterized in that: The invention comprises a polypeptide having the following amino acid sequence: Gly-Ala-Leu-Pro-Ser-Asp-Gly-Glu-Arg-Lys-Gln-Phe-Ser-Gly-Arg-Leu-Ala-Gly-Lys-Ser (SEQ ID NO: 1), and a pharmaceutically acceptable carrier, wherein the molecular weight of the polypeptide is 1500-2000 Daltons and the purity is not less than 95%.

2. A liver-protecting polypeptide preparation according to claim 1, characterized in that: The mass percentage of the polypeptide in the preparation is 1-10%.

3. A hepatoprotective polypeptide preparation according to claim 1, characterized in that: The carrier comprises the following excipients: hydroxypropyl methylcellulose (HPMC), polyvinyl pyrrolidone (PVP), mannitol, microcrystalline cellulose, lactose and polyethylene glycol (PEG).

4. A liver-protecting polypeptide preparation according to claim 3, characterized in that: The mass percentages of the excipients in the preparation are: hydroxypropyl methylcellulose (HPMC) accounts for 15%-25%, polyvinyl pyrrolidone (PVP) accounts for 5%-15%, mannitol accounts for 25%-35%, microcrystalline cellulose accounts for 10%-20%, lactose accounts for 5%-10%, polyethylene glycol (PEG) accounts for 1%-5%, and the balance is purified water.

5. A liver-protecting polypeptide preparation according to claim 1, characterized in that: The dosage form of the preparation is oral solution.

6. A method for preparing a liver-protecting polypeptide preparation, characterized in that: The following steps are involved: Step 1, peptide synthesis: using solid phase synthesis method, using Wang resin as a carrier, sequentially coupling amino acids to synthesize a peptide with a specified sequence, and performing purification after each coupling reaction; Step 2, polypeptide purification: high performance liquid chromatography (HPLC) is used to purify the synthesized polypeptide to ensure that the purity of the polypeptide reaches more than 95%; Step 3, preparation of preparation: mixing the purified polypeptide with the excipients according to the above mass percentage to obtain a uniform mixture; Step 4: Forming the dosage form: Add purified water to the mixture, stir evenly, filter, and then fill it through oral liquid preparation equipment to obtain the final oral liquid preparation.

7. The method for preparing a hepatoprotective polypeptide preparation according to claim 6, characterized in that: The usage amount of the Wang resin is 10%-20% of the total mass of the polypeptide synthesis.

8. The method for preparing a hepatoprotective polypeptide preparation according to claim 6, characterized in that: The high performance liquid chromatography (HPLC) method uses a C18 reverse phase chromatographic column and uses acetonitrile and water as mobile phases for gradient elution, with a flow rate of 1.0 to 1.5 mL / min and a detection wavelength of 220 nm.

9. The method for preparing a hepatoprotective polypeptide preparation according to claim 6, characterized in that: The mixing process is carried out using a high-speed mixer for 20-30 minutes; the filtering process uses a 0.22 micron microporous filter membrane to remove particulate impurities.

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