阅读说明：本技术 一种抗纤维化药物缓释涂层及其制备方法 (Anti-fibrosis drug sustained-release coating and preparation method thereof ) 是由 种铁 杨鹏 于 2020-07-02 设计创作，主要内容包括：本发明公开了一种抗纤维化药物缓释涂层及其制备方法,将蛋白质、蛋白改性剂、添加剂加入去离子水中,混合均匀,并用pH调节剂调节pH至4.0～10.0,制成浸渍液；将抗纤维化药物溶液喷涂固定于医用材料表面,在浸渍液中静置处理即可形成抗纤维化药物缓释涂层。本发明制备方法简便,适用于多种医用器材表面,尤其是生物惰性基材,无需复杂的表面改性技术,通过简单浸涂即可实现。本发明缓释涂层具有良好的可控性,释药稳定,能有效控制药物早期释放率,防止植入早期快速释放所致的毒性作用。此外,本发明缓释涂层的主要成分为蛋白质和天然高分子物质,无毒无刺激,具有良好的生物相容性,在局部药物输送方面具有广阔的应用前景。(The invention discloses an anti-fibrosis drug sustained-release coating and a preparation method thereof, which comprises the steps of adding protein, a protein modifier and an additive into deionized water, uniformly mixing, and regulating the pH to 4.0-10.0 by using a pH regulator to prepare a steeping liquor; the anti-fibrosis drug solution is sprayed and fixed on the surface of the medical material and is kept stand in the immersion liquid to form the anti-fibrosis drug sustained-release coating. The preparation method is simple and convenient, is suitable for the surfaces of various medical devices, especially for biological inert base materials, does not need a complex surface modification technology, and can be realized by simple dip coating. The slow release coating has good controllability and stable drug release, can effectively control the early release rate of the drug and prevent the toxic effect caused by the quick release at the early stage of implantation. In addition, the main components of the slow release coating are protein and natural high molecular substances, so that the slow release coating is non-toxic and non-irritant, has good biocompatibility and has wide application prospect in the aspect of local drug delivery.)

1. A preparation method of an anti-fibrosis drug sustained-release coating is characterized in that:

(1) adding protein, a protein modifier and an additive into deionized water, uniformly mixing, and adjusting the pH to 4.0-10.0 by using a pH regulator to obtain an impregnation solution; the concentration of protein in the impregnation liquid is 0.8-15 mg/mL, the concentration of a protein modifier is 2-20 mg/mL, and the concentration of an additive is 0.4-4 mg/mL;

the protein is any one of lysozyme, bovine serum albumin, insulin, alpha-lactalbumin, human serum albumin, fibrinogen, beta-amyloid protein, Abeta peptide, prion protein, alpha-synuclein, cystatin C, Huntington protein and immunoglobulin light chain;

the protein modifier is any one or more of tris (2-carboxyethyl) phosphine hydrochloride, cysteine and reductive glutathione;

the additive is one or more of sodium carboxymethylcellulose, D-fructose, alginic acid, gelatin, hyaluronic acid and glycerol;

(2) and (2) spraying and fixing the anti-fibrosis drug solution on the surface of the medical material, then placing the medical material fixed with the anti-fibrosis drug in the impregnation liquid obtained in the step (1), standing for 1-3 hours, taking out, washing with deionized water, and drying at room temperature.

2. The method for preparing the anti-fibrosis drug sustained-release coating according to claim 1, characterized in that: in the step (1), the concentration of the protein in the steeping liquor is 2-10 mg/mL, the concentration of the protein modifier is 5-15 mg/mL, and the concentration of the additive is 0.5-2 mg/mL.

3. The method for preparing the anti-fibrosis drug sustained-release coating according to claim 1, characterized in that: in the step (1), a pH regulator is used for regulating the pH value to 5.0-7.0.

4. The method for preparing an anti-fibrotic drug sustained release coating according to claim 1 or 3, characterized in that: in the step (1), the pH regulator is any one or more of sodium hydroxide, sodium carbonate and sodium bicarbonate.

5. The method for preparing the anti-fibrosis drug sustained-release coating according to claim 1, characterized in that: in the step (2), the anti-fibrosis drugs comprise hydrophilic anti-fibrosis drugs and hydrophobic anti-fibrosis drugs, wherein the hydrophilic anti-fibrosis drugs comprise one or a mixture of more of mitomycin C and analogues thereof, bleomycin and derivatives thereof, matrine and halofuginone; the hydrophobic anti-fibrosis drug comprises one or more of paclitaxel drugs, vinca alkaloids drugs, 5-fluorouracil, anthracyclines, non-steroidal anti-inflammatory drugs, fat-soluble antibiotics, camptothecin and derivatives thereof, rapamycin and derivatives thereof, cyclophosphamide and epirubicin hydrochloride.

6. The method for preparing the anti-fibrosis drug sustained-release coating according to claim 5, characterized in that: in the step (2), the hydrophilic anti-fibrosis drug is prepared by taking water or dimethyl sulfoxide as a solvent; the hydrophobic drug solution is prepared by taking a mixed solution of any one or more of methanol, ethanol, isopropanol, dichloromethane, acetone, butanone, tetrahydrofuran and dimethyl sulfoxide as a solvent, and the concentration of the anti-fibrosis drug is 10-50 mg/mL.

7. The method for preparing the anti-fibrosis drug sustained-release coating according to claim 1, characterized in that: in the step (2), the medical material comprises any one of the following materials:

(1) metal material: stainless steel, titanium and its alloys, cobalt-based alloys, nickel-titanium alloys, magnesium and its alloys, zinc and its alloys, iron and its alloys;

(2) inorganic materials: silicon dioxide, titanium dioxide, carbon materials, silicon, titanium dioxide, titanium oxide and titanium nitride;

(3) high polymer material: terylene, polyvinyl alcohol, polyethylene, polytetrafluoroethylene, polyvinyl chloride, polystyrene, polyurethane, polypropylene, polyamide, polycarbonate, polyacrylonitrile, polyacrylic acid and derivatives thereof, polyether ether ketone, silicone rubber, polylactic acid, polyglycolide, polylactide and polycaprolactone;

(4) natural biological material: plastic starch-based materials, sodium alginate, collagen, fibrin, sodium hyaluronate and gelatin;

(5) artificially synthesizing a polypeptide hydrogel material: poly-L-glutamic acid, poly-L-lysine.

8. The method for preparing the anti-fibrosis drug sustained-release coating according to claim 1, characterized in that: in the step (2), the technological parameters of spraying and fixing the anti-fibrosis drug solution on the surface of the medical material mainly comprise: the ultrasonic frequency is 120kHz, the ultrasonic power is 0.2-3.2W, the flow of the ultrasonic spraying solution is 0.5-2 mL/min, the rotating speed of the driving equipment is 300-1200 r/min, the spraying speed is 5-40 mm/s, the spraying frequency is 1-4 times, and the carrier gas pressure is 2-8 Pa.

9. The anti-fibrosis drug sustained-release coating prepared by the method of claim 1.

Technical Field

The invention belongs to the technical field of drug sustained release, and particularly relates to an anti-fibrosis drug sustained release coating and a preparation method thereof.

Background

Tissue and organ damage and repair are one of the difficulties faced by the medical community at present, and the optimal tissue repair mode should be repaired by the same kind of cells to recover the form and function of the tissue. However, the proliferative capacity of each tissue cell in vivo is different, and if the tissue cell cannot be repaired by the same cell, the tissue defect is repaired by proliferation of mesenchymal cells such as fibroblast. Although the repair method can maintain the structural integrity of tissues and organs, the repair method is also accompanied by fibrosis and scar formation of different degrees, which causes the stenosis of hollow organs and lumens and influences the normal physiological functions of human bodies. For benign urethral, biliary and tracheal stenosis, surgical repair and mechanical dilation are the main treatment methods adopted at present, but the treatment effect is severely limited by surgical trauma and complications such as postoperative restenosis. The anti-fibrosis drugs docetaxel, rapamycin and derivatives thereof effectively inhibit the formation of tissue stenosis by regulating cell cycle, extracellular matrix deposition and other ways. However, the oral administration bioavailability of common anti-fibrosis drugs such as docetaxel, rapamycin and the like is low, the individual absorption difference is large, the administration dosage is difficult to control, and the solubilizer is required to be supplemented in the intravenous injection, so that the allergic reaction of the organism is easily caused. More importantly, the whole body administration modes such as oral administration and intravenous injection have low drug selectivity, poor targeting property, large blood concentration fluctuation and obvious whole body toxic and side effects. The local drug delivery system has the advantages of high bioavailability and small systemic adverse reaction, promotes tissue repair, and has wide application prospects in the aspects of tumor resistance, pain relief, inflammation resistance, cardiovascular and cerebrovascular interventional therapy and the like. In recent years, sustained release coatings based on biodegradable polymers have provided new strategies for topical drug delivery. However, acidic degradation products are easy to cause nonbacterial inflammation in vivo and side effects caused by burst release of the drug, so that the practical clinical use value of the acidic degradation products is limited. The drug is combined with the drug-carrying matrix and the surface of the material through covalent bonds, so that the defect of burst release of the drug is effectively overcome, but complex chemical reaction is usually required to form active groups on the surface of the material, and the drug is not suitable for being applied to the surfaces of various medical materials. Therefore, the ideal slow-release coating based on the surface modification of various medical implants has the characteristics of biosafety, universal adhesion, stable drug release and the like.

Disclosure of Invention

Aiming at the problems of obvious early burst release, complex preparation process, poor biocompatibility and the like of the existing slow-release coating technology, the invention provides a slow-release coating which can form a stable amyloid protein film on the surface of anti-fibrosis drug particles through self-assembly by utilizing the characteristic that amyloid protein is assembled and adhered on the solid surface and is used for the surfaces of various medical implants, and a preparation method thereof.

In order to solve the problems, the anti-fibrosis drug sustained-release coating adopted by the invention is prepared by the following method:

1. adding protein, a protein modifier and an additive into deionized water, uniformly mixing, and adjusting the pH to 4.0-10.0 by using a pH regulator to obtain an impregnation solution; the concentration of the protein in the steeping liquid is 0.8-15 mg/mL, the concentration of the protein modifier is 2-20 mg/mL, and the concentration of the additive is 0.4-4 mg/mL.

2. And (3) spraying and fixing the anti-fibrosis drug solution on the surface of the medical material, then placing the medical material fixed with the anti-fibrosis drug in the impregnation liquid obtained in the step (1), standing for 1-3 hours, taking out, washing with deionized water, and drying at room temperature.

The protein is any one of lysozyme, bovine serum albumin, insulin, alpha-lactalbumin, human serum albumin, fibrinogen, beta-amyloid protein, Abeta peptide, prion protein, alpha-synuclein, cystatin C, Huntington protein and immunoglobulin light chain.

The protein modifier is one or more of tris (2-carboxyethyl) phosphine hydrochloride, cysteine and reductive glutathione.

The additive is one or more of sodium carboxymethylcellulose, D-fructose, alginic acid, gelatin, hyaluronic acid and glycerol.

In the step 1, the concentration of the protein in the impregnation liquid is preferably 2-10 mg/mL, the concentration of the protein modifier is preferably 5-15 mg/mL, and the concentration of the additive is preferably 0.5-2 mg/mL.

In the step 1, the pH is further preferably adjusted to 5.0-7.0 by a pH regulator, wherein the pH regulator is one or more of sodium hydroxide, sodium carbonate and sodium bicarbonate.

In the step 2, the anti-fibrosis drugs comprise hydrophilic anti-fibrosis drugs and hydrophobic anti-fibrosis drugs, wherein the hydrophilic anti-fibrosis drugs comprise one or more of mitomycin C and analogues thereof, bleomycin and derivatives thereof, matrine and halofuginone. The hydrophobic anti-fibrosis drug comprises one or more of paclitaxel (paclitaxel and its derivatives, docetaxel and its derivatives), vinca alkaloids (including vinblastine, vincristine, vindesine and vinorelbine), 5-fluorouracil, anthracyclines (adriamycin and epirubicin), non-steroidal anti-inflammatory drugs (ibuprofen, piroxicam, meloxicam, ketorolac, aminopyrine, parecoxib, celecoxib, nimesulide, flurbiprofen, etc.), liposoluble antibiotics (fluoroquinolones, chloromycetin, neomacrolide, tetracycline, clindamycin), camptothecin and its derivatives, rapamycin and its derivatives, cyclophosphamide and epirubicin hydrochloride.

The hydrophilic anti-fibrosis drug is prepared by taking water or dimethyl sulfoxide as a solvent; the hydrophobic drug solution is prepared by taking a mixed solution of any one or more of methanol, ethanol, isopropanol, dichloromethane, acetone, butanone, tetrahydrofuran and dimethyl sulfoxide as a solvent, and the concentration of the anti-fibrosis drug is 10-50 mg/mL.

In the step 2, the medical material includes any one of the following materials:

(1) metal material: stainless steel, titanium and its alloys, cobalt-based alloys, nickel-titanium alloys, magnesium and its alloys, zinc and its alloys, iron and its alloys;

(2) inorganic materials: silicon dioxide, titanium dioxide, carbon materials, silicon, titanium dioxide, titanium oxide and titanium nitride;

(3) high polymer material: terylene, polyvinyl alcohol, polyethylene, polytetrafluoroethylene, polyvinyl chloride, polystyrene, polyurethane, polypropylene, polyamide, polycarbonate, polyacrylonitrile, polyacrylic acid and derivatives thereof, polyether ether ketone, silicone rubber, polylactic acid, polyglycolide, polylactide and polycaprolactone;

(4) natural biological material: plastic starch-based materials, sodium alginate, collagen, fibrin, sodium hyaluronate and gelatin;

(5) artificially synthesizing a polypeptide hydrogel material: poly-L-glutamic acid, poly-L-lysine.

The medical material is cleaned by ethanol and acetone alternately before use, ultrasonically cleaned by deionized water for 10 minutes, and dried by nitrogen.

In the step 2, the technological parameters of spraying and fixing the anti-fibrosis drug solution on the surface of the medical material mainly comprise: the ultrasonic frequency is 120kHz, the ultrasonic power is 0.2-3.2W, the flow of the ultrasonic spraying solution is 0.5-2 mL/min, the rotating speed of the driving equipment is 300-1200 r/min, the spraying speed is 5-40 mm/s, the spraying frequency is 1-4 times, and the carrier gas pressure is 2-8 Pa.

The invention has the following beneficial effects:

1. the preparation method is simple and convenient, is generally suitable for the surfaces of various medical devices, particularly the surfaces of biological inert substrates, does not need complex surface modification technology, and can be realized by simple dip coating. And the coating is stable after the preparation is finished, and is convenient for subsequent use.

2. The invention can effectively control the early release rate of the medicine and prevent the toxic effect caused by the quick release of the medicine in the early period of implantation.

3. The anti-fibrosis drug sustained-release coating has good controllability and stable drug release, can maintain the sustained release of the drug within 10-45 days, and can meet various clinical requirements.

4. The main components of the slow release coating are protein and natural high molecular substances, so the slow release coating is non-toxic and non-irritant, has good biocompatibility and has wide application prospect in the aspect of local drug delivery.

Drawings

FIG. 1 is a scanning electron micrograph of an untreated silicone tube and a rapamycin sustained-release coating coated silicone tube in example 1, wherein a is a surface of the untreated silicone tube, b is a cross section of the rapamycin sustained-release coating coated silicone tube, c is a surface of the rapamycin sustained-release coating coated silicone tube, and d is a partially enlarged view of c.

Figure 2 is an in vitro release profile of the drug-loaded silicone tubing prepared in example 1 and comparative example 1.

FIG. 3 is a scanning electron micrograph of the sustained release coating before and after the flexural fatigue test.

FIG. 4 shows the results of cell proliferation experiments measured by culturing human primary urethral fibroblasts (HUSF) in leaching solution with sustained release coating for 24 hours, 48 hours, and 72 hours.

Detailed Description

The invention will be further described in detail with reference to the following figures and examples, but the scope of the invention is not limited to these examples.