CN111888344A - A kind of preparation method of inflammatory response type intelligent controlled-release drug-loaded composite medical film - Google Patents

Abstract

The invention discloses a preparation method of an inflammation response type intelligent controlled-release drug-loaded composite medical membrane. The intelligent controlled-release composite medical film is prepared by the following method: the method comprises the steps of taking anti-inflammatory drugs indometacin (Indo) and methyl methacrylate (HEMA) as raw materials, synthesizing an indometacin drug-carrying precursor HEIn through an esterification reaction, adding a certain amount of HEIn, HEMA, Azobisisobutyronitrile (AIBN) and N, N' ⁃ Methylene Bisacrylamide (MBA) into Polycaprolactone (PCL) solution for fully mixing, and then carrying out in-situ cross-linking polymerization to prepare the PCL/P (HEMA-HEIn) intelligent controlled-release drug-carrying composite membrane. The controlled-release drug-loaded composite membrane can automatically release drugs in inflammatory environment, and the release amount of the drugs can be adjusted along with the change of the inflammatory environment. The composite membrane has good biocompatibility and inflammation responsiveness, and has wide application prospect in the fields of dressing, hemostatic materials, tissue engineering scaffold materials and the like.

Description

A kind of preparation method of inflammatory response type intelligent controlled-release drug-loaded composite medical film

technical field

The invention relates to a preparation method of an inflammatory response type intelligent controlled-release drug-carrying composite medical film, and belongs to the field of biomedical material preparation.

Background technique

In human diseases, especially in the healing process of chronic diseases such as diabetes, inflammation is a very deadly complication. Therefore, in order to promote wound healing, it is crucial to protect the wound from inflammation. The most commonly used strategy at this stage is to prepare a drug sustained-release medical composite film, but this strategy cannot completely avoid the sudden release of the drug, thus affecting the therapeutic effect. Therefore, a new anti-inflammatory method needs to be found. Commonly used drug-loaded membrane preparation methods include blending, adsorption and covalent bonding. Among them, the drug-loaded composite membrane is prepared by covalently linking the drug with responsive groups, which can not only avoid the sudden release of the drug, but also realize the intelligent controlled release of the drug. At present, there are two main strategies for inflammatory response: one is an anti-oxidative inflammatory response designed based on oxidative stress at the site of inflammation; the other is a pH-sensitive inflammatory response designed based on the pH value of the site of inflammation that is different from that of normal tissues . These two strategies for inflammatory response involve more, however, these two roughly have disadvantages such as poor specificity. Some studies have found that the accumulation of macrophages usually causes a large amount of cholesterol esterase (CE) in the inflammatory site. Using CE enzymes to be sensitive to ester bonds to design ester bond-sensitive inflammatory responses can overcome the clinical differences of inflammatory patients and achieve the purpose of high-efficiency anti-inflammatory. More importantly, CE enzymes can also be depleted through the inflammatory response of drug ester bonds, thereby reducing the loss of medical membranes in the inflammatory environment and enhancing the therapeutic effect.

Indomethacin (Indometacin, Indo) is a non-steroidal anti-inflammatory drug, also known as indomethacin. Indomethacin contains carboxyl groups in its structure, which can react with hydroxyl groups to form ester bonds. Hydroxyethyl methacrylate (HEMA) contains hydroxyl groups in its molecular structure, which can react with indomethacin to form ester bonds and generate drug precursors for inflammatory responses. In addition, HEMA also contains active carbon-carbon double bonds, which can further polymerize and functionalize the drug precursor on the substrate, thereby realizing intelligent controlled release in response to inflammation. Therefore, the present invention designs and obtains a drug precursor HEIn containing indomethacin, and then compound the drug precursor HEIn with PCL to achieve functionalization, and prepare an inflammatory response type intelligent controlled-release drug-loaded composite membrane, which not only The method is innovative, and the prepared medical composite membrane has good biocompatibility and inflammatory response.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a composite membrane with good biocompatibility and good inflammatory response for the field of biomedical materials, which can be applied to the fields of wounds, hemostatic materials, tissue engineering scaffold materials and the like.

Object of the present invention can be realized by following preparation technology, and its preparation method steps are as follows:

(1) Preparation of indomethacin-loaded precursor HEIn: Weigh a certain amount of indomethacin and dissolve it in a certain amount of mixed solvent of acetone (acetone) and dichloromethane (DCM). A certain amount of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) was weighed and dissolved in a certain amount of mixed solvent of water and acetone. A certain amount of 4-dimethylaminopyridine (DMAP) was weighed and dissolved in a certain amount of dichloromethane. Weigh a certain amount of HEMA and dissolve it in a certain amount of dichloromethane. The above prepared solution is added to the reaction kettle, and the reaction is fully stirred at a certain temperature for a certain period of time. Subsequently, the reactant was concentrated, separated and purified, and finally the product HEIn was obtained;

(2) Preparation of PCL/P(HEME-HEIn) intelligent controlled-release drug-loaded composite film: Weigh a certain amount of PCL and dissolve it in a certain amount of dimethylformamide (DMF). A certain amount of HEIn, HEMA, initiator (AIBN) and cross-linking agent (MBA) were added to the PCL solution, and the solution was fully stirred at room temperature to form a homogeneous solution. The prepared solution is poured into the mold, and the mold is placed in an oven at a certain temperature to react for a certain period of time. After the reaction is completed, the mold is cooled, and the drug-loaded film is taken out from the mold;

(3) The above-mentioned composite medical membrane was immersed in physiological saline, PBS buffer and deionized water successively to remove the solvent remaining in the composite medical membrane, and then sterilized by γ-rays generated at a dose of 6~30KGy/h60Co after drying. bacteria, form packaging, and obtain an inflammation-responsive intelligent controlled-release composite medical film.

In above-mentioned preparation process, the molar ratio of HEMA described in step (1) and indomethacin is 100:1～1:100, and the molar ratio of EDC and indomethacin is 100:1～1: 100, the molar ratio of DMAP and indomethacin is 100:1～1:100, and temperature of reaction is-4～100 ℃, and the reaction times is 1～96 h. The PCL molecular weight described in step (2) is 14000-80000 g/mol, and the PCL concentration is 4%-40% (w/w). The mass ratio of PCL to monomer (the sum of HEIn and HEMA) is 100:1~1:100, the mass ratio of HEIn and HEMA is 100:1~1:100, and the mass ratio of MBA and HEMA is 1:1000~ 1:1, the mass ratio of AIBN and MBA is 1:1000~1:1, the oven temperature is 30~150 °C, and the reaction time is 0.5~72 h.

Compared with the prior art, the present invention has the following advantages:

(1) Design and synthesis of a new drug precursor HEIn for smart drug controlled release, which contains active double bonds in its structure, which can be widely used as an agent for the controlled release of inflammatory smart response drugs;

(2) The experimental conditions of HEIn are mild, the energy consumption is low, and it is easy to implement;

(3) In-situ cross-linking polymerization is used to realize the functionalization of polycaprolactone for inflammatory response, which is simple and time-saving, with less material and low cost;

(4) The inflammatory response intelligent controlled-release polycaprolactone medical composite membrane prepared by the present invention has good biocompatibility and inflammatory response, and has excellent comprehensive performance.

Detailed ways

The present invention will be specifically described below through implementation. It is necessary to point out that this embodiment is only used to further illustrate the present invention, and should not be construed as a limitation on the protection scope of the invention. Some non-essential improvements and adjustments have been made to the content.

Example 1

(1) Preparation of indomethacin-loaded precursor HEIn: 716 g of indomethacin was weighed and dissolved in a certain amount of mixed solvent of acetone (acetone) and dichloromethane (DCM). Weigh 400 g of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and dissolve it in a certain amount of mixed solvent of water and acetone. Weigh 300 g of 4-dimethylaminopyridine (DMAP) and dissolve it in a certain amount of dichloromethane. Weigh 260 g of HEMA and dissolve it in a certain amount of dichloromethane. The above prepared solution was added to the reaction kettle, and the reaction was fully stirred at room temperature for 24 h. Subsequently, the reactant was concentrated, separated and purified, and finally the product HEIn was obtained;

(2) Preparation of PCL/P(HEME-HEIn) intelligent controlled-release drug-loaded composite film: Weigh 180 g of PCL, dissolve it in a certain amount of dimethylformamide (DMF), and prepare 15 % (w/ w) solution. Add 60 g HEIn, 120 g HEMA, 18 g initiator (AIBN) and 144 g cross-linking agent (MBA) to the PCL solution, and stir well at room temperature to form a homogeneous solution. The prepared solution was poured into a mold, and the mold was placed in an oven at 70 °C for 12 h. After the reaction is completed, the mold is cooled, and the drug-loaded film is taken out from the mold;

(3) The above-mentioned composite medical membrane was immersed in physiological saline, PBS buffer and deionized water successively to remove the solvent remaining in the composite medical membrane, and then sterilized by γ-rays generated at a dose of 6~30KGy/h60Co after drying. bacteria, form packaging, and obtain an inflammation-responsive intelligent controlled-release composite medical film.

Example 2

(1) Preparation of indomethacin-loaded precursor HEIn: 720 g of indomethacin was weighed and dissolved in a certain amount of mixed solvent of acetone (acetone) and dichloromethane (DCM). Weigh 200 g of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and dissolve it in a certain amount of mixed solvent of water and acetone. Weigh 100 g of 4-dimethylaminopyridine (DMAP) and dissolve it in a certain amount of dichloromethane. Weigh 50 g of HEMA and dissolve it in a certain amount of dichloromethane. The above prepared solution was added to the reaction kettle, and the reaction was fully stirred for 12 h at 30 °C. Subsequently, the reactant was concentrated, separated and purified, and finally the product HEIn was obtained;

(2) Preparation of PCL/P(HEME-HEIn) intelligent controlled-release drug-loaded composite film: Weigh 270 g of PCL, dissolve it in a certain amount of dimethylformamide (DMF), and prepare 24 % (w/ w) solution. Add 100 g HEIn, 100 g HEMA, 100 g initiator (AIBN) and 600 g cross-linking agent (MBA) to the PCL solution, and stir well at room temperature to form a homogeneous solution. The prepared solution was poured into a mold, and the mold was placed in an oven at 60 °C for 8 h. After the reaction is completed, the mold is cooled, and the drug-loaded film is taken out from the mold;

(3) The above-mentioned composite medical membrane was immersed in physiological saline, PBS buffer and deionized water successively to remove the solvent remaining in the composite medical membrane, and then sterilized by γ-rays generated at a dose of 6~30KGy/h60Co after drying. bacteria, form packaging, and obtain an inflammation-responsive intelligent controlled-release composite medical film.

Example 3

(1) Preparation of indomethacin-loaded precursor HEIn: 800 g of indomethacin was weighed and dissolved in a certain amount of mixed solvent of acetone (acetone) and dichloromethane (DCM). Weigh 200 g of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and dissolve it in a certain amount of mixed solvent of water and acetone. Weigh 200 g of 4-dimethylaminopyridine (DMAP) and dissolve it in a certain amount of dichloromethane. Weigh 400 g of HEMA and dissolve it in a certain amount of dichloromethane. The above-mentioned prepared solution was added to the reaction kettle, and the reaction was fully stirred for 10 h at 40 °C. Subsequently, the reactant was concentrated, separated and purified, and finally the product HEIn was obtained;

(2) Preparation of PCL/P(HEME-HEIn) intelligent controlled-release drug-loaded composite film: Weigh 300 g of PCL, dissolve it in a certain amount of dimethylformamide (DMF), and prepare 20 % (w/ w) solution. Add 120 g HEIn, 120 g HEMA, 40 g initiator (AIBN) and 700 g cross-linking agent (MBA) to the PCL solution and stir well at room temperature to form a homogeneous solution. The prepared solution was poured into a mold, and the mold was placed in an oven at 100 °C for 7 h. After the reaction is completed, the mold is cooled, and the drug-loaded film is taken out from the mold;

(3) The above-mentioned composite medical membrane was immersed in physiological saline, PBS buffer and deionized water successively to remove the solvent remaining in the composite medical membrane, and then sterilized by γ-rays generated at a dose of 6~30KGy/h60Co after drying. bacteria, form packaging, and obtain an inflammation-responsive intelligent controlled-release composite medical film.

Claims (3)

1. A preparation method of an inflammation response type intelligent controlled-release drug-loaded composite medical membrane is characterized by comprising the following steps:

(1) preparing an indometacin drug-loaded precursor HEIn: weighing a certain amount of indometacin, and dissolving the indometacin in a certain amount of mixed solvent of acetone (acetone) and Dichloromethane (DCM); weighing a certain amount of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), and dissolving in a certain amount of mixed solvent of water and acetone; weighing a certain amount of 4-Dimethylaminopyridine (DMAP), and dissolving in a certain amount of dichloromethane; weighing a certain amount of HEMA, and dissolving in a certain amount of dichloromethane; adding the prepared solution into a reaction kettle, and fully stirring and reacting for a certain time at a certain temperature; then concentrating the reactant, separating and purifying to finally obtain a product HEIn;

(2) preparing a PCL/P (HEME-HEIn) intelligent controlled-release drug-loaded composite membrane: weighing a certain amount of PCL, and dissolving in a certain amount of Dimethylformamide (DMF); adding a certain amount of HEIn, HEMA, initiator (AIBN) and cross-linking agent (MBA) into the PCL solution, and fully stirring at room temperature to obtain a uniform solution; pouring the prepared solution into a mold, and placing the mold in a drying oven at a certain temperature for reaction for a certain time; after the reaction is finished, cooling the mold, and taking the drug-loaded film out of the mold;

(3) and (3) sequentially soaking the composite medical membrane in normal saline, PBS (phosphate buffer solution) and deionized water to remove the solvent remained in the composite medical membrane, drying, sterilizing by gamma rays generated by 6-30 KGy/h60Co, forming and packaging to obtain the inflammation response type intelligent controlled-release composite medical membrane.

2. The preparation method of the inflammation-responsive intelligent controlled-release drug-loaded composite medical membrane of claim 1, wherein the feeding molar ratio of HEMA to indomethacin in the step (1) is 100: 1-1: 100, the feeding molar ratio of EDC to indomethacin is 100: 1-1: 100, the feeding molar ratio of DMAP to indomethacin is 100: 1-1: 100, the reaction temperature is-4-100 ℃, and the reaction time is 1-96 h.

3. The preparation method of the inflammation-responsive intelligent controlled-release drug-loaded composite medical membrane of claim 1, wherein the molecular weight of PCL in the step (2) is 14000-80000 g/mol, and the concentration of PCL is 4% -40% (w/w); the mass ratio of PCL to monomers (the sum of HEIn and HEMA) is 100: 1-1: 100, the mass ratio of HEIn to HEMA is 100: 1-1: 100, the mass ratio of MBA to HEMA is 1: 1000-1: 1, the mass ratio of AIBN to MBA is 1: 1000-1: 1, the oven temperature is 30-150 ℃, and the reaction time is 0.5-72 h.