Amino acid-based polyester amide / polyglycolide fiber membrane and preparation method and application thereof

Abstract

The present application relates to a kind of preparation method of amino acid-based polyester amide / polyglycolide fiber membrane.The fiber membrane is that protein-based hypoglycemic drug is added to 0.2~0.5g / ml amino acid-based polyester amide solution to obtain drug-loaded nanocomposite, then is added to 0.2~0.5g / ml polyglycolide solution, is prepared by electrospinning technology, wherein the volume ratio of amino acid-based polyester amide drug-loaded nanocomposite polyglycolide solution is 0.5~2:5.The fiber membrane of the present application has very high porosity, has good adhesion when implanted into patient's body, prevents the risk of postoperative adhesion, can effectively promote the rapid healing of postoperative wound of diabetic patients;In addition, the preparation method of the fiber membrane is simple, has good application in the preparation of diabetic wound repair material.

Description

Technical Field

[0001] This invention relates to a poly(ethylene lactide) fiber membrane, its preparation method and application, and more specifically, to an amino acid-based polyesteramide / poly(ethylene lactide) fiber membrane, its preparation method and application. Background Technology

[0002] Postoperative adhesions are abnormal adhesions that occur during tissue healing. These adhesions typically occur between tissues near the surgical incision and sometimes even involve organs far from the incision. Postoperative adhesions not only restrict normal organ movement but can also lead to pain, functional impairment, and in some cases, bowel obstruction or other serious complications. Diabetic patients, due to their high blood sugar levels, experience suppressed body repair mechanisms, resulting in significantly slower wound healing, a process that can take weeks or even months, placing a heavy psychological and financial burden on patients and their families. Currently, the difficulty in healing diabetic wounds and the lack of effective treatments have made this a challenging and urgent clinical and scientific problem. Summary of the Invention

[0003] The purpose of this invention is to provide an amino acid-based polyesteramide / poly(ethylene glycol) fiber membrane and its preparation method to solve problems such as poor wound healing and postoperative adhesion risk in diabetic patients.

[0004] This invention is implemented as follows: A method for preparing an amino acid-based polyesteramide / poly(lactic acid) fiber membrane includes the following steps: First, a protein-based hypoglycemic drug is added to a 0.2-0.5 g / ml amino acid-based polyesteramide solution to obtain a drug-loaded nanocomposite, wherein the mass ratio of the protein-based hypoglycemic drug to the amino acid-based polyesteramide is 0.1-1:10; then, the above-mentioned drug-loaded nanocomposite is added to a 0.2-0.5 g / ml poly(lactic acid) solution, wherein the volume ratio of the drug-loaded nanocomposite to the poly(lactic acid) solution is 0.5-2:5; and the amino acid-based polyesteramide / poly(lactic acid) fiber membrane is obtained by electrospinning.

[0005] The preparation method of the amino acid-based polyester amide is as follows: (1) using amino acids and diols as raw materials, adding end-group protectants, and esterifying at 120℃~140℃ to obtain amino acid-based sulfonylbenzene or xylene salt ester monomers; (2) reacting nitrophenol, triethylamine, and diacyl chloride in an organic solvent at -80~0℃ for 1~4h, gradually raising the temperature to room temperature and continuing the reaction for 4~12h, then precipitating with sufficient distilled water, washing, recrystallizing with acetonitrile 3 times, and vacuum drying for 48h to obtain di-p-nitrophenyl ester of dicarboxylic acid. Amino acid sulfonate monomer and di-p-nitrophenyl ester of dicarboxylic acid were added to solvent N,N-dimethylformamide (DMF) at a mass ratio of 1:1. The mixture was stirred until homogeneous and heated to 80°C. Triethylamine, in an amount equal to 2.2 times the molar volume of the amino acid sulfonate monomer, was added dropwise under rapid stirring. The reaction was carried out at 80°C for 24 h. The product was then precipitated with glacial ethyl acetate, dissolved in dichloromethane, and precipitated again with ethyl acetate. This purification process was repeated three times. Finally, the product was dried in a vacuum drying oven at 50°C for 48 h to obtain amino acid-based polyesteramide.

[0006] The amino acid is any one of phenylalanine, alanine, arginine, and lysine, with phenylalanine being preferred.

[0007] The diol is any one of propylene glycol, butanediol, hexanediol, polyethylene glycol 200, or polyethylene glycol 400, with hexanediol being preferred.

[0008] The diacyl chloride is any one of oxaloyl chloride, adipicoyl chloride, octanoyl chloride, and sebacate, with octanoyl chloride being preferred.

[0009] The protein-based hypoglycemic drug is any one of insulin, exenatide-4, and liraglutide, with exenatide-4 being preferred.

[0010] The solvent used in the amino acid-based polyester amide solution is dimethyl sulfoxide, N,N-dimethylformamide, etc., preferably dimethyl sulfoxide.

[0011] The solvent used in the poly(lactic acid) lactide solution is either chloroform / dimethyl sulfoxide or chloroform / N,N-dimethylformamide in a volume ratio of 1:1, with chloroform / dimethyl sulfoxide being preferred.

[0012] The poly(glycolic acid) is a carboxyl-terminated lactide-glycolic acid copolymer, wherein the lactide molar percentage content is 70-80% and the intrinsic viscosity is 60-80 ml / g.

[0013] The conditions for electrospinning are as follows: spinning voltage of 10-55kV, syringe capacity of 10ml, 21# needle, injection speed of 0.8-1.6ml / h, and receiving distance of 10-30cm.

[0014] The present invention also provides an amino acid-based polyesteramide / poly(lactic acid) fiber membrane prepared by the above preparation method and its application in the preparation of postoperative wound repair materials for diabetic patients.

[0015] This invention synthesizes an amino acid-based polyesteramide with good electropositive properties using amino acids, diols, and diacyl chlorides as raw materials. The invention utilizes electrostatic interactions to prepare a nanocomposite of the positively charged amino acid-based polyesteramide and a protein-based hypoglycemic drug, which is then compounded with negatively charged polyethylene glycol (PEG). Electrospinning is then used to obtain an amino acid-based polyesteramide / PEG fiber membrane carrying the hypoglycemic drug.

[0016] The amino acid-based polyesteramide / poly(lactic acid) fiber membrane of this invention has excellent biocompatibility, is lightweight, soft, has a high specific surface area, high porosity, and good air permeability, which can meet the needs of cell growth and is suitable as a postoperative anti-adhesion material. In addition, it can encapsulate different types of protein-based hypoglycemic drugs that promote wound healing, achieving localized, efficient, and sustained hypoglycemic effects. When implanted into the patient, it has excellent conformability and can effectively promote rapid wound healing in diabetic patients after surgery, preventing postoperative adhesion risks and other problems.

[0017] The preparation method of the amino acid-based polyesteramide / poly(lactic acid) fiber membrane of this invention is simple and easy to promote on a large scale. It has good application prospects and broad development space in postoperative anti-adhesion isolation materials and diabetic wound repair materials. Attached Figure Description

[0018] Figure 1 The contact angle of the amino acid-based polyesteramide / poly(lactic acid) composite fiber membrane prepared according to the present invention.

[0019] Figure 2 The microstructure of the amino acid-based polyesteramide / poly(lactic acid) composite fiber membrane prepared according to the present invention is shown.

[0020] Figure 3 The cumulative drug release curve of the amino acid-based polyesteramide / poly(lactic acid) composite fiber membrane prepared according to the present invention. Detailed Implementation

[0021] The present invention will be further illustrated below with reference to embodiments. These embodiments are for illustrative purposes only and do not limit the scope of protection of the present invention in any way. Processes and methods not described in detail in the following embodiments are conventional methods known in the art. Unless otherwise specified, the reagents and materials used in the following embodiments are commercially available.

[0022] This invention combines positively charged amino acid-based polyesteramide, negatively charged poly(lactic acid) lactide, and protein-based hypoglycemic drugs through electrostatic interaction to prepare an amino acid-based polyesteramide / poly(lactic acid) fiber membrane.

[0023] The preparation method of the positively charged amino acid-based polyester amide is as follows: (1) Synthesis of bisphenylalanine sulfonate monomer: 0.02 mol phenylalanine, 0.05 mol p-toluenesulfonic acid monohydrate and 0.01 mol hexanediol were added to the organic solvent toluene and heated to 130 °C for 10 h to obtain crude p-bisphenylalanine sulfonate. The crude product was washed with distilled water several times, filtered and vacuum dried for 48 h to obtain bisphenylalanine sulfonate.

[0024] (2) Synthesis of di-p-nitrophenyl ester monomer of dicarboxylic acid: 0.02 mol of p-nitrophenol and 0.02 mol of triethylamine were added to 40 ml of acetone at -20℃ and stirred evenly for later use; octanoyl chloride was added to 20 ml of acetone at -20℃ and mixed evenly, and then added dropwise to the above mixture. After stirring at -20℃ for 2 h, the temperature was gradually raised to room temperature and the reaction continued for 10 h. Then, the mixture was precipitated with sufficient distilled water, washed, and recrystallized three times with acetonitrile. After vacuum drying for 48 h, di-p-nitrophenyl ester of octanoic acid was obtained with a yield of 85%.

[0025] (3) Preparation of phenylalanine-based polyester amide polymer: (a) Diphenylalanine sulfonate monomer and di-p-nitrophenyl octanoate in a mass ratio of 1:1 were added to the solvent N,N-dimethylformamide (DMF), stirred evenly and heated to 80°C. Triethylamine in an amount equal to 2.2 times the amount of diphenylalanine sulfonate monomer was added dropwise under rapid stirring. The reaction was carried out at 80°C for 24 h. The product was then precipitated with ethyl acetate, dissolved in dichloromethane, and precipitated again with ethyl acetate. The purification was repeated 3 times. Finally, the product was dried in a vacuum drying oven at 50°C for 48 h to obtain phenylalanine-based polyester amide polymer.

[0026] The polyethylene glycol used in the following examples is a carboxyl-terminated lactide-glycolic acid copolymer, wherein the lactide molar percentage is 70-80% and the intrinsic viscosity is 60-80 ml / g. The preparation method of this polyethylene glycol is as follows: According to the molar ratio of 3:1, dried lactide and glycolide were added to ampoules. 0.015% stannous octoate and 0.01% octanoic acid were added according to the total mass of lactide and glycolide. The ampoules were sealed under vacuum and placed in an oil bath at 140~160℃ for 24 hours. Then, the mixture was dissolved in dichloromethane, precipitated with ethanol, and dried under vacuum at 50℃ for 24 hours to obtain poly(glycolic acid) lactide polymer.

[0027] Example 1 10g of phenylalanine-based polyesteramide was added to dimethyl sulfoxide to prepare a 0.5g / ml solution. Then, 4g of exenatide-4 was added to the phenylalanine-based polyesteramide solution to obtain an amino acid-based polyesteramide drug-loaded nanocomposite. 10g of polyethylene glycol lactide was dissolved in a chloroform / dimethyl sulfoxide mixed solvent to prepare a 0.5g / ml polyethylene glycol lactide solution. 0.5ml of the amino acid-based polyesteramide drug-loaded nanocomposite and 5ml of the polyethylene glycol lactide solution were stirred evenly to obtain a drug-loaded amino acid-based polyesteramide / polyethylene glycol lactide electrospinning solution. Electrospinning was performed using the following conditions: spinning voltage 20kV, syringe capacity 10ml, 21# needle, injection speed 1.2ml / h, and receiving distance 20cm, to obtain an amino acid-based polyesteramide / polyethylene glycol lactide composite fiber membrane with a thickness of 128mm, an elongation at break of 430%, and a solvent residue of 0.08%. The static contact angle of this fiber membrane is as follows: Figure 1 As shown in (A), its value is 117.2°. After soaking in PBS buffer solution at 37°C and pH 7.0 for 24 h, the fiber membrane retained 96.10% of its size, exhibiting excellent extensibility and dimensional stability, which is beneficial for handling and use. The microstructure of the fiber membrane is shown in Figure [Figure Number]. Figure 2 As shown in (A), the fiber membrane is composed of countless nanoscale fibers randomly stacked together, exhibiting a high porosity. The drug release behavior of this fiber membrane is as follows... Figure 3 (A) The release of hypoglycemic drugs gradually over time can maintain local blood glucose stability.

[0028] Drug release determination method: Weigh the above drug-loaded amino acid-based polyesteramide / polyethylene lactide fiber membrane and add it to PBS buffer solution with pH 7.0. Incubate it in a shaker at 37°C. Take 2 mL of sample at the set time and add 2 mL of fresh PBS buffer solution. Then test its concentration with a UV spectrophotometer and calculate the cumulative drug release result.

[0029] Example 2 10g of phenylalanine-based polyesteramide was added to dimethyl sulfoxide to prepare a 0.2g / ml solution. Then, 1g of exenatide-4 was added to the phenylalanine-based polyesteramide solution to obtain an amino acid-based polyesteramide drug-loaded nanocomposite. 10g of polyethylene glycol lactide was dissolved in a chloroform / dimethyl sulfoxide mixed solvent to prepare a 0.2g / ml polyethylene glycol lactide solution. 2ml of the amino acid-based polyesteramide drug-loaded nanocomposite and 5ml of the polyethylene glycol lactide solution were stirred evenly to obtain a drug-loaded amino acid-based polyesteramide / polyethylene glycol lactide electrospinning solution. Electrospinning was performed using the following parameters: spinning voltage 30kV, syringe capacity 10ml, 21# needle, injection speed 0.8ml / h, and receiving distance 18cm, to obtain a drug-loaded amino acid-based polyesteramide / polyethylene glycol lactide composite fiber membrane. The membrane thickness was 124mm, elongation at break was 510%, and solvent residue was 0.1%. The static contact angle of this fiber membrane is as follows: Figure 1 As shown in (B), its value is 108.1°. After soaking in PBS buffer solution at 37°C and pH 7.0 for 24 hours, the fiber membrane retained 99.8% of its size, exhibiting excellent extensibility and dimensional stability, which is beneficial for handling and use. The microstructure of this fiber membrane is shown in Figure [Figure number missing]. Figure 2 As shown in (B), it has excellent porosity. The drug release from this fiber membrane is as follows... Figure 3 (B) The hypoglycemic drug is released gradually over time, which can maintain local blood glucose stability. The method for determining the drug release is the same as in Example 1.

[0030] Example 3 10g of phenylalanine-based polyesteramide was added to dimethyl sulfoxide to prepare a 0.5g / ml solution. Then, 2g of exenatide-4 was added to the phenylalanine-based polyesteramide solution to obtain an amino acid-based polyesteramide drug-loaded nanocomposite. 10g of polyethylene glycol lactide was dissolved in a chloroform / dimethyl sulfoxide mixed solvent to prepare a 0.5g / ml polyethylene glycol lactide solution. 1ml of the amino acid-based polyesteramide drug-loaded nanocomposite and 5ml of the polyethylene glycol lactide solution were stirred evenly to obtain a drug-loaded amino acid-based polyesteramide / polyethylene glycol lactide electrospinning solution. Electrospinning was performed using the following parameters: spinning voltage 55kV, syringe capacity 10ml, 21# needle, injection speed 0.8ml / h, and receiving distance 30cm, to obtain a drug-loaded amino acid-based polyesteramide / polyethylene glycol lactide composite fiber membrane. The fiber membrane thickness was 130mm, the elongation at break was 380%, and the solvent residue was 0.05%. The static contact angle of this fiber membrane is as follows: Figure 1 As shown in (C), its value is 103.8°. After soaking in PBS buffer solution at 37°C and pH 7.0 for 24 hours, the fiber membrane retained 97.8% of its size, exhibiting excellent extensibility and dimensional stability, which is beneficial for handling and use. The microstructure of this fiber membrane is shown in Figure [Figure number missing]. Figure 2As shown in (C), it has excellent porosity. The drug release from this fiber membrane is as follows... Figure 3 (C) The hypoglycemic drug is released gradually over time, maintaining local blood glucose stability. The method for determining the drug release is the same as in Example 1.

[0031] Example 4 10g of phenylalanine-based polyesteramide was added to dimethyl sulfoxide to prepare a 0.5g / ml solution. Then, 4g of exenatide-4 was added to the phenylalanine-based polyesteramide solution to obtain an amino acid-based polyesteramide drug-loaded nanocomposite. 10g of polyethylene glycol lactide was dissolved in a chloroform / dimethyl sulfoxide mixed solvent to prepare a 0.5g / ml polyethylene glycol lactide solution. 2ml of the amino acid-based polyesteramide drug-loaded nanocomposite and 5ml of the polyethylene glycol lactide solution were stirred evenly to obtain a drug-loaded amino acid-based polyesteramide / polyethylene glycol lactide electrospinning solution. Electrospinning was performed using the following conditions: spinning voltage 18kV, syringe capacity 10ml, 21# needle, injection speed 1.6ml / h, and receiving distance 10cm, to obtain a drug-loaded amino acid-based polyesteramide / polyethylene glycol lactide composite fiber membrane. The fiber membrane thickness was 140mm, the elongation at break was 550%, and the solvent residue was 0.15%. The static contact angle of this fiber membrane is as follows: Figure 1 As shown in (D), its value is 135.1°. After soaking in PBS buffer solution at 37°C and pH 7.0 for 24 h, the fiber membrane retained 95.8% of its size, exhibiting excellent extensibility and dimensional stability, which is beneficial for handling and use. The microstructure of this fiber membrane is shown in Figure [Figure number missing]. Figure 2 As shown in (D), it has excellent porosity. The drug release behavior of this fiber membrane is as follows... Figure 3 (D) The hypoglycemic drug is released gradually over time, maintaining local blood glucose stability. The method for determining the drug release is the same as in Example 1.

Claims

1. A method for preparing an amino acid-based polyesteramide / poly(lactic acid) fiber membrane, characterized in that, The process includes the following steps: First, a protein-based hypoglycemic drug is added to a 0.2-0.5 g / ml amino acid-based polyesteramide solution to obtain a drug-loaded nanocomposite, wherein the mass ratio of the protein-based hypoglycemic drug to the amino acid-based polyesteramide is 0.1-1:10; then, the above drug-loaded nanocomposite is added to a 0.2-0.5 g / ml polyethylene glycol lactide solution, wherein the volume ratio of the drug-loaded nanocomposite to the polyethylene glycol lactide solution is 0.5-2:5; and an amino acid-based polyesteramide / polyethylene glycol lactide fiber membrane is obtained by electrospinning.

2. The method for preparing the amino acid-based polyesteramide / poly(lactic acid) fiber membrane according to claim 1, characterized in that: The protein-based hypoglycemic drug is any one of insulin, exenatide-4, or liraglutide.

3. The method for preparing the amino acid-based polyesteramide / poly(lactic acid) fiber membrane according to claim 1, characterized in that: The solvent used in the amino acid-based polyesteramide solution is dimethyl sulfoxide or N,N-dimethylformamide.

4. The method for preparing the amino acid-based polyesteramide / poly(lactic acid) fiber membrane according to claim 1, characterized in that: The solvent used for the poly(ethylene glycol) lactide solution is a 1:1 volume ratio mixture of chloroform and dimethyl sulfoxide or a 1:1 volume ratio mixture of chloroform and N,N-dimethylformamide.

5. The method for preparing the amino acid-based polyesteramide / poly(lactic acid) fiber membrane according to claim 1, characterized in that: The poly(glycolic acid) is a carboxyl-terminated lactide-glycolic acid copolymer, wherein the lactide molar percentage content is 70-80% and the intrinsic viscosity is 60-80 ml / g.

6. The method for preparing the amino acid-based polyesteramide / poly(lactic acid) fiber membrane according to claim 1, characterized in that: The conditions for electrospinning are as follows: spinning voltage of 10-55kV, syringe capacity of 10ml, 21# needle, injection speed of 0.8-1.6ml / h, and receiving distance of 10-30cm.

7. The method for preparing the amino acid-based polyesteramide / poly(lactic acid) fiber membrane according to claim 1, characterized in that, The method for preparing the amino acid-based polyester amide is as follows: amino acid-based sulfonate esters are prepared using amino acids and diols as raw materials; di-p-nitrophenyl ester of diacid is prepared using nitrophenol and diacyl chloride as raw materials; the amino acid-based sulfonate ester monomer and di-p-nitrophenyl ester of diacid are reacted at a mass ratio of 1:1 to obtain amino acid-based polyester amide.

8. The method for preparing the amino acid-based polyesteramide / poly(lactic acid) fiber membrane according to claim 7, characterized in that: The amino acid is any one of phenylalanine, alanine, arginine, and lysine; the diol is any one of propylene glycol, butanediol, hexanediol, polyethylene glycol 200, or polyethylene glycol 400; and the diacyl chloride is any one of oxaloyl chloride, adipicoyl chloride, octanoyl chloride, or sebacyl chloride.

9. The amino acid-based polyesteramide / poly(lactic acid) fiber membrane prepared by the preparation method according to any one of claims 1 to 8.

10. The application of the amino acid-based polyesteramide / poly(lactic acid) fiber membrane according to claim 9 in the preparation of diabetic wound repair materials.

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