CN104474595A - Production method of dilayer compounding coated esophageal stent with esophageal cancer treatment assisting function - Google Patents

Abstract

The invention relates to a preparation method of an esophageal stent with a double-layer composite film covering function of adjuvant treatment of esophageal cancer. In the present invention, a dense inner polycaprolactone film is prepared on the surface of the Ni-Ti memory esophageal stent by the method of solution coating; Prepare the solution of the outer layer of fibrous membrane, and use the Ni-Ti memory esophageal stent with an inner layer of polycaprolactone dense film on the surface as the rotating receiver of the electrospinning device, and use the electrospinning technology to obtain the solution obtained from the outer layer of fiber membrane The fibrous membrane is spun on the inner polycaprolactone dense film to obtain a double-layer composite coating with the function of treating esophageal cancer; the double-layer composite coating is composed of an outer fibrous membrane and an inner polycaprolactone dense It is compounded when the solvent has not evaporated completely, and it fits tightly. The invention realizes the controlled release of medicine while improving the cell compatibility of the membrane material; the invention has simple process, easy operation and low cost.

Description

Preparation method of double-layer composite film-covered esophageal stent with function of adjuvant treatment of esophageal cancer

technical field

The invention relates to the preparation of a multifunctional esophageal stent covering, in particular to a preparation method of a double-layer composite covering esophageal stent with the function of assisting the treatment of esophageal cancer.

Background technique

Esophageal cancer is a common digestive tract tumor. About 310,000 people die from esophageal cancer every year in the world, of which 160,000 occur in China. The typical symptom of esophageal cancer is progressive dysphagia, which will lead to gradual weight loss of the patient, directly reduce the quality of life of the patient, and even cause death. Since the early symptoms of esophageal cancer are not obvious, when the diagnosis is made, the cancer cells may be in the middle or late stage or have already spread. In addition, some patients have large lesions, so only about 50% of the patients are suitable for surgical resection. Therefore, in order to alleviate the patient's difficulty in eating and drinking, implanting an esophageal stent is a very important palliative therapy.

Esophageal stents are divided into bare stents and covered stents coated with special membrane materials. Compared with the bare stent, the covered stent can effectively prevent the growth of hyperplastic tissue into the stent, and it is easy to remove after the esophageal stricture is relieved. Therefore, the covering material on the covered stent needs to have good mechanical properties and biocompatibility . On this basis, adding anticancer drugs to the membrane of the covered stent can not only play the role of a general covered stent, but also locally release the drug at a fixed point to achieve a more rapid and effective auxiliary killing effect on cancer cells. , which is far more targeted and has a higher utilization rate than general intravenous injection of anticancer drugs.

Combined with the current medical situation in our country, there is an urgent need for a covered stent that can be implanted in the lesion area and has adjuvant treatment for esophageal cancer. Pulling the wound in the middle causes secondary damage, and at the same time achieves the effect of adjuvant therapy for esophageal cancer.

Contents of the invention

The purpose of the present invention is to provide a preparation method of a double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer in view of the deficiencies of the prior art.

The present invention firstly prepares a solution for preparing a dense inner layer film, and prepares a dense inner layer film on the surface of the Ni-Ti memory esophageal stent through the method of solution coating; The mixed solution obtained in the organic solvent is the solution for preparing the outer layer of fibrous membrane. The Ni-Ti memory esophageal stent with a dense inner layer on the surface is used as the rotating receiver of the electrospinning equipment. The fibrous membrane obtained by the solution is spun on the inner dense membrane, thereby obtaining a double-layer composite membrane with the function of treating esophageal cancer; It is compounded when volatilization is complete, and fits tightly. The double-layer composite coating of the esophageal stent with the function of adjuvant treatment of esophageal cancer obtained by the preparation method is composed of an inner dense membrane and an outer fibrous membrane, and the inner dense membrane can adhere well It is attached to a metal stent, and the outer fibrous membrane is made of a mixed solution containing organic polymers and anticancer drugs by electrospinning technology. The soft fiber material of the outer fibrous membrane has weak mechanical properties and can Avoid causing secondary damage to the wound during the removal of the stent; in addition, it can also control the release of anticancer drugs.

The preparation method of the double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer of the present invention comprises the following steps:

(1) Preparation and preparation of polycaprolactone (PCL) inner film solution: dissolve polycaprolactone in an organic solvent, prepare a solution with a concentration of 10-20% w/v, stir at room temperature (generally stir for about 3 hours) ) evenly;

(2) Preparation of inner polycaprolactone (PCL) dense film: pour the solution obtained in step (1) into a petri dish, place the compressed Ni-Ti memory esophageal stent in the petri dish and roll until the polycaprolactone (PCL) The lactone solution evenly covers the surface of the compressed Ni-Ti memory esophageal stent; the compressed Ni-Ti memory esophageal stent is released to restore the compressed Ni-Ti memory esophageal stent to its original length, and the Ni-Ti memory esophageal stent The inner polycaprolactone dense film is obtained on the surface;

(3) Prepare the mixed solution for preparing the outer fiber membrane: dissolve the organic polymer material in an organic solvent, prepare a solution with a concentration of 9 to 15% w/v, add anticancer drugs, and stir at room temperature (usually for 12 hours) left and right) evenly to obtain a mixed solution;

(4) Preparation of double-layer composite coating: inject the mixed solution obtained in step (3) into the syringe of the electrospinning equipment, add a stainless steel needle with a diameter of 0.3 to 0.7 mm on the front end of the syringe, and obtain The Ni-Ti memory esophageal stent restored to the original length is a rotating receiver; electrospinning is carried out under the conditions of a voltage of 12-25kv, a solution flow rate of 3-8ml/h, and a distance of 10-25cm from the rotating receiver to the stainless steel needle. , thus on the surface of the Ni-Ti memory esophageal stent, a double-layer composite film consisting of an inner layer of polycaprolactone dense film and an outer layer of fibrous film containing organic polymer materials and anticancer drugs is obtained; vacuum drying at room temperature ( Generally, the vacuum drying time at room temperature is 24 to 48 hours), and the residual organic solvent is removed to obtain a double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer. The double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer is stored in a dry box at 4°C.

Adding the anticancer drug described in step (3), the amount of the anticancer drug added is that the mass ratio of the mass of the anticancer drug to the organic polymer material in the solution with a concentration of 9 to 15% w/v is 3 to 3. 15% to add anticancer drugs.

The thickness of the outer fiber membrane of the double-layer composite coating is 30-150 μm.

The inner dense film of the double-layer composite film has a thickness of 10-80 μm.

The organic polymer material is selected from one or more of lactic acid-glycolic acid copolymer (PLGA), polycaprolactone (PCL), and gelatin (GT). The weight average molecular weight of the PCL is 50,000-200,000.

The weight average molecular weight of the PLGA is 50,000-200,000.

The anticancer drug is selected from one of paclitaxel, doxorubicin, 5-fluorouracil and the like.

The organic solvent is selected from one or two of DMF, acetone, trifluoroethanol, hexafluoroisopropanol, tetrahydrofuran, chloroform, dichloromethane, and dimethyl sulfoxide.

The electrostatic spinning equipment is a multi-spinner electrostatic spinning machine.

Beneficial effects of the present invention: the double-layer composite film-covered esophageal stent prepared by the present invention has the function of adjuvant treatment of esophageal cancer. The inner dense film can be well adhered to the metal stent, and the soft fiber The mechanical properties of the material are weak, and no secondary damage will be caused to the wound during the removal of the esophageal stent. The double-layer composite coating is loaded with anticancer drugs, and by adjusting the ratio of organic polymer materials, the controlled release of the drug is also achieved while improving the cell compatibility of the membrane material; and the double-layer composite coating material Anti-cancer drugs have less damage to normal cells while maintaining a sustained killing effect on cancer cells. The preparation method of the double-layer composite film-coated esophageal stent with the function of adjuvant treatment of esophageal cancer has simple process, easy operation and low cost, and has wide application prospects.

Detailed ways

The present invention will be described in further detail below through specific examples, but it should not be interpreted as limiting the protection scope of the present invention. All modifications, replacements, and changes based on the above-mentioned technical ideas and utilizing common technical knowledge and conventional means in the art All belong to the scope of the present invention.

Example 1

(1) Preparation and preparation of polycaprolactone (PCL, weight average molecular weight is 100,000) inner film solution: dissolve PCL in tetrahydrofuran, prepare a solution with a concentration of 10% w/v, and stir at room temperature for about 3 hours;

(2) Preparation of inner PCL dense film: pour the solution obtained in step (1) into a petri dish, place the compressed Ni-Ti memory esophageal stent in the petri dish and roll until the PCL solution is in the compressed Ni-Ti The surface of the Ti memory esophageal stent is evenly covered; take out and release the compressed Ni-Ti memory esophageal stent, so that the compressed Ni-Ti memory esophageal stent returns to its original length, and the thickness on the surface of the Ni-Ti memory esophageal stent is about 10 μm The inner polycaprolactone dense film;

(3) Prepare the mixed solution for preparing the outer fiber membrane: PLGA (the weight average molecular weight is 80,000), PCL (the weight average molecular weight is 200,000) and GT, and the mass ratio of PLGA:PCL:GT is 10:2.5: The ratio of 5 was dissolved in hexafluoroisopropanol together, and a solution with a concentration of 11% w/v was prepared, paclitaxel was added, and stirred at room temperature for about 12 hours to obtain a well-mixed mixture; the amount of paclitaxel added was based on the paclitaxel The ratio of mass to the total mass of PLGA, PCL and GT solute in the solution of 11% w/v is 10% to add paclitaxel;

(4) Preparation of double-layer composite film: inject the homogeneously mixed solution obtained in step (3) into the syringe of the multi-spinneret electrospinning machine, and add a stainless steel needle with a diameter of 0.5 mm to the front end of the syringe to The Ni-Ti memory esophageal stent restored to the original length obtained in step (2) is a rotating receiver; electrospinning is performed under the conditions that the voltage is 20kv, the solution flow rate is 5ml/h, and the distance between the rotating receiver and the stainless steel needle is 20cm. Thus, a double-layer composite film consisting of an inner polycaprolactone dense film and an outer fibrous film containing PLGA, PCL, GT and paclitaxel was obtained on the surface of the Ni-Ti memory esophageal stent, and the thickness of the outer fibrous film It is about 150 μm; vacuum-dried at room temperature for 48 hours to remove the residual organic solvent, and obtain a double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer. The double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer is stored in a dry box at 4°C.

Example 2

(1) Preparation and preparation of polycaprolactone (PCL, weight average molecular weight is 100,000) inner film solution: dissolve PCL in tetrahydrofuran, prepare a solution with a concentration of 15% w/v, and stir at room temperature for about 3 hours;

(2) Preparation of inner PCL dense film: pour the solution obtained in step (1) into a petri dish, place the compressed Ni-Ti memory esophageal stent in the petri dish and roll until the PCL solution is in the compressed Ni-Ti The surface of the Ti memory esophageal stent is evenly covered; take out and release the compressed Ni-Ti memory esophageal stent, so that the compressed Ni-Ti memory esophageal stent returns to its original length, and the thickness on the surface of the Ni-Ti memory esophageal stent is about 50 μm The inner polycaprolactone dense film;

(3) Prepare the mixed solution for preparing the outer fiber membrane: PLGA (the weight average molecular weight is 200,000), PCL (the weight average molecular weight is 100,000) and GT, and the mass ratio of PLGA:PCL:GT is 10:2.5: The ratio of 1 is dissolved in hexafluoroisopropanol together, and a solution with a concentration of 9% w/v is prepared, paclitaxel is added, and stirred at room temperature for about 12 hours to obtain a well-mixed mixture; the amount of paclitaxel added is based on the amount of paclitaxel The ratio of mass to the total mass of PLGA, PCL and GT in a solution of 9% w/v is 3% to add paclitaxel;

(4) Preparation of double-layer composite film: inject the homogeneously mixed solution obtained in step (3) into the syringe of the multi-spinneret electrospinning machine, add a stainless steel needle with a diameter of 0.3 mm on the front end of the syringe, and The Ni-Ti memory esophageal stent restored to the original length obtained in step (2) is a rotating receiver; electrospinning is performed under the conditions that the voltage is 12kv, the solution flow rate is 3ml/h, and the distance between the rotating receiver and the stainless steel needle is 10cm. Thus, a double-layer composite film consisting of an inner polycaprolactone dense film and an outer fibrous film containing PLGA, PCL, GT and paclitaxel was obtained on the surface of the Ni-Ti memory esophageal stent, and the thickness of the outer fibrous film It is about 100 μm; vacuum-dried at room temperature for 48 hours to remove the residual organic solvent, and obtain a double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer. The double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer is stored in a dry box at 4°C.

Example 3

(1) Preparation and preparation of polycaprolactone (PCL, weight average molecular weight is 50,000) inner film solution: dissolve PCL in methylene chloride, prepare a solution with a concentration of 20% w/v, and stir at room temperature for about 3 hours ;

(2) Preparation of inner PCL dense film: pour the solution obtained in step (1) into a petri dish, place the compressed Ni-Ti memory esophageal stent in the petri dish and roll until the PCL solution is in the compressed Ni-Ti The surface of the Ti memory esophageal stent is uniformly covered, and the compressed Ni-Ti memory esophageal stent is taken out and released, so that the compressed Ni-Ti memory esophageal stent returns to its original length, and the thickness on the surface of the Ni-Ti memory esophageal stent is about 80 μm The inner polycaprolactone dense film;

(3) Prepare the mixed solution for preparing the outer fiber membrane: PLGA (the weight average molecular weight is 50,000), PCL (the weight average molecular weight is 200,000) and GT, and the mass ratio of PLGA:PCL:GT is 10:2.5: The ratio of 1 is dissolved in hexafluoroisopropanol together, and a solution with a concentration of 15% w/v is prepared, and paclitaxel is added, and stirred at room temperature for about 12 hours to obtain a uniformly mixed mixture; the amount of paclitaxel is based on the amount of paclitaxel The ratio of mass to the total mass of PLGA, PCL and GT solute in the solution of 15% w/v is 3% to add paclitaxel;

(4) Preparation of double-layer composite film: inject the homogeneously mixed solution obtained in step (3) into the syringe (the specification is 5ml) of the multi-spinneret electrospinning machine, and add a diameter of 0.7mm to the front end of the syringe. The stainless steel needle, and the Ni-Ti memory esophageal stent returned to the original length obtained in step (2) is used as a rotating receiver; under the condition that the voltage is 25kv, the solution flow rate is 8ml/h, and the distance between the rotating receiver and the stainless steel needle is 25cm Electrospinning was carried out to obtain a double-layer composite film consisting of an inner polycaprolactone dense film and an outer fibrous film containing PLGA, PCL, GT and paclitaxel on the surface of the Ni-Ti memory esophageal stent. The thickness of the fibrous membrane is about 50 μm; it is vacuum dried at room temperature for 48 hours to remove the residual organic solvent, and a double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer is obtained. The double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer is stored in a dry box at 4°C.

Example 4

(1) Preparation and preparation of polycaprolactone (PCL, weight average molecular weight is 200,000) inner layer membrane solution: dissolve PCL in tetrahydrofuran, prepare a solution with a concentration of 10% w/v, and stir at room temperature for about 3 hours;

(2) Preparation of inner PCL dense film: pour the solution obtained in step (1) into a petri dish, place the compressed Ni-Ti memory esophageal stent in the petri dish and roll until the PCL solution is in the compressed Ni-Ti The surface of the Ti memory esophageal stent is uniformly covered, and the compressed Ni-Ti memory esophageal stent is taken out and released, so that the compressed Ni-Ti memory esophageal stent returns to its original length, and the thickness on the surface of the Ni-Ti memory esophageal stent is about 30 μm The inner polycaprolactone dense film;

(3) Prepare the mixed solution for preparing the outer fiber membrane: PLGA (the weight average molecular weight is 100,000) and PCL (the weight average molecular weight is 100,000) are dissolved in the PLGA:PCL mass ratio of 4:1. In a mixed solvent composed of tetrahydrofuran, chloroform and DMF (volume ratio 2:1:2), prepare a solution with a concentration of 15% w/v, add 5-fluorouracil, and stir at room temperature for about 12 hours to obtain a well-mixed mixture. solution; the amount of 5-fluorouracil added is to add 5-fluorouracil according to the ratio of the mass of 5-fluorouracil to the total mass of PLGA and PCL solute in the solution of 15% w/v;

(4) Preparation of double-layer composite film: inject the homogeneously mixed solution obtained in step (3) into the syringe of the multi-spinneret electrospinning machine, and add a stainless steel needle with a diameter of 0.5 mm to the front end of the syringe to The Ni-Ti memory esophageal stent restored to the original length obtained in step (2) is a rotating receiver; electrospinning is performed under the conditions that the voltage is 20kv, the solution flow rate is 5ml/h, and the distance between the rotating receiver and the stainless steel needle is 25cm. Thus, a double-layer composite film consisting of an inner polycaprolactone dense film and an outer fibrous film containing PLGA, PCL and 5-fluorouracil was obtained on the surface of the Ni-Ti memory esophageal stent, and the thickness of the outer fibrous film It is about 100 μm; vacuum-dried at room temperature for 48 hours to remove the residual organic solvent, and obtain a double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer. The double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer is stored in a dry box at 4°C.

Example 5

(1) Preparation and preparation of polycaprolactone (PCL, weight average molecular weight is 200,000) inner film solution: dissolve PCL in tetrahydrofuran, prepare a solution with a concentration of 12% w/v, and stir at room temperature for about 3 hours;

(2) Preparation of inner PCL dense film: pour the solution obtained in step (1) into a petri dish, place the compressed Ni-Ti memory esophageal stent in the petri dish and roll until the PCL solution is in the compressed Ni-Ti The surface of the Ti memory esophageal stent is uniformly covered, and the compressed Ni-Ti memory esophageal stent is taken out and released, so that the compressed Ni-Ti memory esophageal stent returns to its original length, and the thickness on the surface of the Ni-Ti memory esophageal stent is about 30 μm The inner polycaprolactone dense film;

(3) Preparation of the mixed solution for preparing the outer fiber membrane: PLGA (with a weight-average molecular weight of 150,000) was dissolved in hexafluoroisopropanol, prepared into a solution with a concentration of 12% w/v, added paclitaxel, and stirred at room temperature for 12 About an hour, obtain the mixed liquid of mixing uniformly; Its adding amount of paclitaxel is to add paclitaxel according to the quality ratio of the quality of paclitaxel and the PLGA solute in the solution that concentration is 12% w/v is 10%;

(4) Preparation of double-layer composite film: inject the homogeneously mixed solution obtained in step (3) into the syringe of the multi-spinneret electrospinning machine, and add a stainless steel needle with a diameter of 0.5 mm to the front end of the syringe to The Ni-Ti memory esophageal stent restored to the original length obtained in step (2) is a rotating receiver; electrospinning is performed under the conditions that the voltage is 20kv, the solution flow rate is 5ml/h, and the distance between the rotating receiver and the stainless steel needle is 20cm. Thus, on the surface of the Ni-Ti memory esophageal stent, a double-layer composite film consisting of an inner layer of PCL dense film and an outer layer of fibrous film containing PLGA and paclitaxel was obtained, and the thickness of the outer layer of fibrous film was about 150 μm; After drying for 26 hours, the residual organic solvent was removed, and a double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer was obtained. The double-layer composite film-covered esophageal stent with the function of adjuvant treatment of esophageal cancer is stored in a dry box at 4°C.

Claims (10)

1. have a preparation method for the Esophageal Stent of the two-layer compound overlay film of auxiliary treatment esophageal carcinoma function, it is characterized in that, described preparation method comprises the steps:

(1) prepare polycaprolactone inner layer film solution: be dissolved in by polycaprolactone in organic solvent, be mixed with the solution that concentration is 10 ~ 20%w/v, stirring at room temperature is even;

(2) internal layer polycaprolactone dense film is prepared: solution step (1) obtained falls in culture dish, the Ni-Ti of compression memory Esophageal Stent is placed in this culture dish roll, until the surface uniform that polycaprolactone solution remembers Esophageal Stent at the Ni-Ti of this compression covers; The Ni-Ti discharging this compression remembers Esophageal Stent, and make the Ni-Ti of compression remember Esophageal Stent and return to original length, the surface remembering Esophageal Stent at Ni-Ti obtains internal layer polycaprolactone dense film;

(3) prepare the mixed liquor of outer layer fiber film: be dissolved in by high-molecular organic material in organic solvent, be mixed with the solution that concentration is 9 ~ 15%w/v, add cancer therapy drug, stirring at room temperature is even, obtains mixed liquor;

(4) two-layer compound overlay film is prepared: mixed liquor step (3) obtained injects the syringe of electrospinning device, add that in the front end of syringe diameter is the rustless steel syringe needle of 0.3 ~ 0.7mm, the Ni-Ti returning to original length obtained with step (2) remembers Esophageal Stent for rotating receiver; Be 12 ~ 25kv at voltage, solution flow rate is 3 ~ 8ml/h, rotating receiver distance rustless steel syringe needle is carry out electrostatic spinning under the condition of 10 ~ 25cm, the two-layer compound overlay film that the fibrous membrane obtaining containing high-molecular organic material and cancer therapy drug by internal layer polycaprolactone dense film and skin on the surface that Ni-Ti remembers Esophageal Stent is formed; Dry at room temperature in vacuo, remove residual organic solvent, obtain the Esophageal Stent of the two-layer compound overlay film with auxiliary treatment esophageal carcinoma function.

2. preparation method according to claim 1, it is characterized in that: add cancer therapy drug described in step (3), to be the mass values being high-molecular organic material in the solution of 9 ~ 15%w/v according to the quality of cancer therapy drug and concentration be for the addition of its cancer therapy drug 3 ~ 15% to add cancer therapy drug.

3. preparation method according to claim 1, is characterized in that: the thickness of the outer layer fiber film of described two-layer compound overlay film is 30 ~ 150 μm.

4. preparation method according to claim 1, is characterized in that: the thickness of the inner compact layer film of described two-layer compound overlay film is 10 ~ 80 μm.

5. preparation method according to claim 1 and 2, is characterized in that: described high-molecular organic material is selected from one or more in poly lactic coglycolic acid, polycaprolactone, gelatin.

6. preparation method according to claim 1, is characterized in that: the weight average molecular weight of described polycaprolactone is 50,000 ~ 200,000.

7. preparation method according to claim 5, is characterized in that: the weight average molecular weight of described polycaprolactone is 50,000 ~ 200,000.

8. preparation method according to claim 5, is characterized in that: the weight average molecular weight of described poly lactic coglycolic acid is 50,000 ~ 200,000.

9. preparation method according to claim 1 and 2, is characterized in that: described cancer therapy drug is selected from the one in paclitaxel, amycin, 5-fluorouracil.

10. preparation method according to claim 1, is characterized in that: described organic solvent be selected from DMF, acetone, trifluoroethanol, hexafluoroisopropanol, oxolane, chloroform, dichloromethane, dimethyl sulfoxide one or both.