DRUG-LOADED TISSUE ADHESIVE FILM AND PREPARATION METHOD THEREFOR

Abstract

The present invention provides a drug-loaded tissue adhesive film, comprising alternately superposed cationic layers and anionic layers, at least one of the cationic layers and the anionic layers being a drug layer, or at least one of the cationic layers and the anionic layers containing a drug with charges. The provided drug-loaded tissue adhesive film has good tissue adhesiveness, biocompatibility, degradable absorption, and stability, and the physical and chemical properties of the drug-loaded tissue adhesive film can be adjusted by adjusting material compositions.

Claims

1. A drug-loaded tissue adhesive film, comprising alternately superposed cationic layers and anionic layers, wherein at least one of the cationic layers and the anionic layers is a drug layer, or at least one of the cationic layers and the anionic layers contains a drug with charges.

2. The drug-loaded tissue adhesive film according to claim 1, characterized in that, when at least one of the cationic layers and the anionic layers is the drug layer, the cationic layer and/or the anionic layer used as the drug layer is the drug with charges, the cationic layer not used as the drug layer contains a carrier material with a cationic group, and the anionic layer not used as the drug layer contains a carrier material with an anionic group; and when at least one of the cationic layers and the anionic layers contains the drug with charges, the cationic layer contains a carrier material with a cationic group and the anionic layer contains a carrier material with an anionic group.

3. The drug-loaded tissue adhesive film according to claim 2, characterized in that the carrier material with a cationic group is one or a combination of an organic high-molecular polymer with a cationic group, a polysaccharide with a cationic group, a polypeptide with a cationic group, a protein with a cationic group, and a cationic liposome.

4. The drug-loaded tissue adhesive film according to claim 2, characterized in that the carrier material with an anionic group is one or a combination of an organic high-molecular polymer with an anionic group, a polysaccharide with an anionic group, a polypeptide with an anionic group, a protein with an anionic group, and an anionic liposome.

5. The drug-loaded tissue adhesive film according to claim 2, characterized in that the carrier material with a cationic group and the carrier material with an anionic group are biocompatible materials.

6. The drug-loaded tissue adhesive film according to claim 1, characterized in that the cationic layer presents positive charges on the whole and the anionic layer presents negative charges on the whole.

7. The drug-loaded tissue adhesive film according to claim 1, characterized in that the drug with charges is a drug complex.

8. A method for preparing the drug-loaded tissue adhesive film according to claim 1, comprising the following step: alternately depositing cationic layers and anionic layers on a substrate to prepare the tissue adhesive film.

9. Application of a tissue adhesive film to preparation of a drug carrier material, the tissue adhesive film comprises alternately superposed cationic layers and anionic layers.

10. The application according to claim 9, characterized in that at least one of the cationic layers and the anionic layers is a drug layer, or at least one of the cationic layers and the anionic layers is used for containing a drug with charges.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0050] FIG. 1 illustrates a schematic view in a gel electrophoresis experiment of siRNA in embodiment 12.

[0051] FIG. 2 illustrates schematic views of cell transfection tests in embodiment 13.

[0052] FIG. 3 illustrates a schematic view of relative fluorescence intensity in embodiment 13.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0053] The embodiments of the present invention will be described below through specific examples. One skilled in the art can easily understand other advantages and effects of the present invention according to the content disclosed in the description. The present invention may also be implemented or applied through other different specific implementation modes. Various modifications or variations may be made to all details in the description based on different points of view and applications without departing from the spirit of the present invention.

[0054] Before the embodiments of the present invention are further described, it should be understood that the protective scope of the present invention is not limited to the following specific implementation solutions; and it should be further understood that the terms used in the embodiments of the present invention are used for describing specific implementation solutions instead of limiting the protective scope of the present invention; and in the description and claims of the present invention, unless otherwise clearly pointed out, the singular forms “a,” “an,” and “the” are intended to include the plural forms as well.

[0055] When numerical value ranges are given in the embodiments, it should be understood that, unless otherwise indicated, two endpoints of each numerical value range and any numerical values between the two endpoints are selectable. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by one skilled in the art. In addition to the specific methods, devices and materials used in the embodiments, as known by one skilled in the art to the prior art and recorded in the present invention, any methods, devices and materials in the prior art similar or equal to the methods, devices and materials in the embodiments of the present invention may also be used to implement the present invention.

[0056] Unless otherwise stated, the experiment methods, detection methods and preparation methods disclosed in the present invention adopt conventional molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell cure, recombinant DNA technique and common techniques in the related art. These techniques have already been perfectly described in the current literatures. For details, refer to Sambrook, et al, MOLECULAR CLONING: A LABORATORY MANUAL, Second edition, Cold Spring Harbor Laboratory Press, 1989 and Third edition, 2001; Ausubel, et al, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, 1987 and periodic updates; the series METHODS IN ENZYMOLOGY, Academic Press, San Diego; Wolfe, CHROMATIN STRUCTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998; METHODS IN ENZYMOLOGY, Vol. 304, Chromatin (P. M. Wassarman and A. P. Wolfe, eds.), Academic Press, San Diego, 1999; and METHODS IN MOLECULAR BIOLOGY, Vol. 119, Chromatin Protocols (P. B. Becker, ed.), Humana Press, Totowa, 1999, etc.

Embodiment 1

[0057] In an aseptic bench, a culture dish with diameter of 12 cm was prepared, a silicon dice film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (1 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (1 mg/ml carboxymethyl chitosan, 0.15 mol/L NaCl, pH=6) were respectively prepared; part of solution B was taken and added with a small interfering nucleic acid drug (eGFP-siRNA (sense: 5′-GGCACAAGCUGGAGUACAAUU-3′; antisense: 5′-UUGUACUCCAGCUUGUGCCUU-3′, 20 ug/ml), a cell targeting factor (hyaluronic acid with a molecular weight smaller than 20000 Daltons, 10 ug/ml) and a targeting polypeptide (1*10.sup.−4 mg/ml) to prepare solution C, wherein an amino acid sequence of the targeting polypeptide was: valine-glycine-valine-alanine-proline-glycine; the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution C was instantaneously coated at high pressure into the culture dish, and drying was performed to form a film; then the solution A was instantaneously coated at high pressure and washing was performed by the water for injection; then the solution B was instantaneously coated at high pressure, washing was performed by water for injection, the solution A and the solution B were alternately coated in this way repeatedly; then the solution A was instantaneously coated at high pressure and washing was performed by water for injection; and finally the solution C was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and then dried to obtain an implantable tissue adhesive film loading the small interfering nucleic acid drug.

Embodiment 2

[0058] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml alginic acid with a molecular weight of 60000-80000, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a small interfering nucleic acid drug (the same as embodiment 1, 20 ug/ml) and a cell targeting factor (hyaluronic acid with a molecular weight smaller than 20000 Daltons, 10 ug/ml), and the solution A was added with a targeting polypeptide (1*10.sup.−4 mg/ml), wherein an amino acid sequence of the targeting polypeptide was: isoleucine-lysine-valine-alanine-valine; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 160 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain an implantable tissue adhesive film loading the small interfering nucleic acid drug.

Embodiment 3

[0059] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution A was added with a small interfering nucleic acid cationic liposome (for a method for preparing a cationic liposome, please refer to Anionic Liposome-Cationic Liposome Complex Mediated Gene Transfection, Journal of Pharmaceutical Practice, 2011 (29): 4, the small interfering nucleic acid is the same as that in embodiment 1, the added amount of the small interfering nucleic acid cationic liposome is 0.5 mg/ml, the drug loading rate of the small interfering nucleic acid is 21.7%), and uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: arginine-glycine-aspartic acid; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 180 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain an implantable tissue adhesive film loading the small interfering nucleic acid drug.

Embodiment 4

[0060] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a small interfering nucleic acid anionic liposome (for a preparation method, please refer to Anionic Liposome-Cationic Liposome Complex Mediated Gene Transfection, Journal of Pharmaceutical Practice, 2011 (29): 4, the small interfering nucleic acid is the same as that in embodiment 1, the added amount of the small interfering nucleic acid anionic liposome is 0.5 mg/ml, the drug loading rate of the small interfering nucleic acid is 12.4%), uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: valine-glycine-valine-alanine-proline-glycine; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 180 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain an implantable tissue adhesive film loading the small interfering nucleic acid drug.

Embodiment 5

[0061] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution A was added with a small interfering nucleic acid cationic liposome (for a preparation method, refer to embodiment 3, the concentration being 0.5 mg/ml) and a targeting polypeptide (1*10.sup.−4 mg/ml), and uniform stirring was performed, wherein an amino acid sequence of the targeting polypeptide was: arginine-glycine-aspartic acid; the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 200 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the small interfering nucleic acid drug.

Embodiment 6

[0062] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml alginic acid with a molecular weight of 60000-80000, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a Sorafenib hyaluronic acid microsphere (the concentration is 1.5 mg/ml, the diameter of the microsphere is 10-16 μm, the drug loading rate is 7.58%), and the solution A was added with a targeting polypeptide (1*10.sup.−4 mg/ml), wherein an amino acid sequence of the targeting polypeptide was: isoleucine-lysine-valine-alanine-valine; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 30 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain an implantable tissue adhesive film loading the Sorafenib drug.

Embodiment 7

[0063] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a Sunitinib sodium alginate microsphere (the concentration is 1.0 mg/ml, the diameter of the microsphere is 10-15 μm, the drug loading rate is 6.28%), the solution A was then added with a targeting polypeptide (1*10.sup.−4 mg/ml) and uniform stirring was performed, wherein an amino acid sequence of the targeting polypeptide was: arginine-glycine-aspartic acid; the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 50 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the Sunitinib drug.

Embodiment 8

[0064] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a ganglioside sodium alginate microsphere (the concentration is 1.0 mg/ml, the diameter of the microsphere is 0.15-0.26 μm, the drug loading rate being 7.3%), and uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: valine-glycine-valine-alanine-proline-glycine; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 30 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain an implantable tissue adhesive film loading the ganglioside drug.

Embodiment 9

[0065] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with an antibacterial peptide (1.5 mg/ml); the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 80 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the antibacterial peptide drug.

Embodiment 10

[0066] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with interleukin-2 (0.5 mg/ml); the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 50 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the interleukin-2 drug.

Embodiment 11

[0067] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution A was added with an Afatinib cationic PEG nanometer microsphere (the concentration is 0.5 mg/ml), and uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: arginine-glycine-aspartic acid; the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 200 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the Afatinib drug.

Embodiment 12

[0068] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution A was added with an Imatinib cationic PEG nanometer microsphere (the concentration is 0.8 mg/ml), and uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: arginine-glycine-aspartic acid; the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 200 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the Imatinib drug.

Embodiment 13

[0069] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with an Axitinib anionic PEG nanometer microsphere (the concentration is 0.8 mg/ml), and uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: arginine-glycine-aspartic acid; the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 200 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the Axitinib drug.

Embodiment 14

[0070] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution A was added with a Ceritinib cationic PEG nanometer microsphere (the concentration is 0.8 mg/ml), and uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: arginine-glycine-aspartic acid; the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 200 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the Ceritinib drug.

Embodiment 15

[0071] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with fluorouracil (1.0 mg/ml); the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 100 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the fluorouracil drug.

Embodiment 16

[0072] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml alginic acid with a molecular weight of 60000-80000, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a Pertuzumab drug (20 ug/ml), and the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 100 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain an implantable tissue adhesive film loading the Pertuzumab drug.

Embodiment 17

[0073] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a Pertuzumab drug (20 ug/ml); the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 100 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the Pertuzumab drug.

Embodiment 18

[0074] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a Lidamycin (0.5 mg/ml, macromolecular protein antitumor antibiotics); the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 80 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the Lidamycin drug.

Embodiment 19

[0075] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a vascular endothelial growth factor (VEGF) (0.5 mg/ml), and uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: valine-glycine-valine-alanine-proline-glycine; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 80 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain a tissue adhesive film loading the vascular endothelial growth factor (VEGF) drug.

Embodiment 20

[0076] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with an epidermal growth factor (EGF) (0.5 mg/ml), and uniform stirring was performed; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 100 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain a tissue adhesive film loading the epidermal growth factor (EGF) drug.

Embodiment 21

[0077] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a nerve growth factor (NGF) (0.5 mg/ml); the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 50 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the nerve growth factor (NGF) drug.

Embodiment 22

[0078] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a fibroblast growth factor (FGF) (0.5 mg/ml); the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 80 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain a tissue adhesive film loading the fibroblast growth factor (FGF) drug.

Embodiment 23

[0079] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml alginic acid with a molecular weight of 60000-80000, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a heparin drug (1.5 mg/ml); the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 100 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain a tissue adhesive film loading the heparin drug.

Embodiment 24

[0080] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a Lycium barbarum polysaccharide drug (1.0 mg/ml); the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 100 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain a tissue adhesive film loading the Lycium barbarum polysaccharide drug.

Embodiment 25

[0081] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with lentinan (0.5 mg/ml); the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 80 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried treatment was performed to obtain an implantable tissue adhesive film loading the lentinan drug.

Embodiment 26

[0082] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with pachymaran (0.5 mg/ml), and uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: valine-glycine-valine-alanine-proline-glycine; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 80 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain a tissue adhesive film loading the pachymaran drug.

Embodiment 27

[0083] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with Polyporus polysaccharide (0.5 mg/ml), and uniform stirring was performed; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 100 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain a tissue adhesive film loading the Polyporus polysaccharide drug.

Embodiment 28

[0084] The drug-loaded tissue adhesive film drug delivery systems obtained in embodiments 1-27 were subjected to biocompatibility evaluation detection, and showed excellent biocompatibility, specifically as follows:

[0085] 1. Cytotoxicity Test:

[0086] Reference/technical standard: GB/T 16886.5-2003

[0087] Cell line: L-929 cells (mouse fibroblasts)

[0088] Culture solution: DMEM with 10% (v/v) calf serum

[0089] Blank control: the same-batch cell culture solution

[0090] Negative control: high density polyethylene (see GB/T16886 Cytotoxicity Test Standard)

[0091] Positive control: 5 g/L phenol solution

[0092] Extraction medium: the same-batch DMEM with no calf serum

[0093] Extraction time: 24 h

[0094] Test sample extraction ratio: 1 g/5 ml

[0095] Test method: extract test (MTT method)

[0096] At 27° C., 5% CO.sub.2 blank control, negative control, positive control and test sample extract contacted with adherently grown cells, culture was performed for 72 h, then MTT solution was added and incubation was performed for 4 h. After resorption, DMSO was added, the absorbance of each group at a wavelength of 630 nm was measured through a spectrophotometer, and the relative proliferation rate of the cells was calculated.

[0097] Results: cytotoxicity of the test sample: level 0

[0098] Conclusion: according to GB/T 16886.5-2003, the test sample is not cytotoxic.

[0099] 2. Intradermal Stimulation Test

[0100] Reference/technical standard: GB/T 16886. 10-2005

[0101] Test animal: healthy New Zealand rabbit

[0102] Extract medium: 0.9% sodium chloride injection

[0103] Test sample: material extract

[0104] Negative control: the same-batch extraction medium

[0105] Contact route: intradermal injection

[0106] Evaluation index: erythema and edema reaction degree at 24 h, 48 h and 72 h

[0107] Result: there is no erythema and edema reaction in the local and peripheral skin tissues at 24 h, 48 h, and 72 h after injection, and the skin reaction on the test side is not greater than the skin reaction on the control side.

[0108] Conclusion: according to GB/T 16886. 10-2005, the test sample has no intradermal stimulation.

[0109] 3. Acute Systemic Toxicity Test

[0110] Reference/technical standard: GB/T 16886. 11-1997/ASTM F 750

[0111] Test animals: healthy mice

[0112] Extraction medium: 0.9% sodium chloride injection

[0113] Test sample: Material extract

[0114] Negative control: the same-batch extract medium

[0115] Contact route: tail vein injection

[0116] Evaluation index: general state of animals, toxicity performance and number of dead animals at 4 h, 24 h, 48 h and 72 h

[0117] Results: the response of the animals in the test sample group is not greater than that in the negative control group during the observation period of 72 h.

[0118] Conclusion: referring to GB/T 16886. 11-1997/ASTM F 750, the results of the acute systemic toxicity test of the test sample are in line with requirements.

[0119] 4. Hemolysis Test

[0120] Reference/technical standard: GB/T 16886. 4-2003/GB/T 16175-1996

[0121] Test animals: healthy New Zealand rabbits

[0122] Diluted anticoagulant rabbit blood preparation: fresh anticoagulant rabbit blood+0.9% sodium chloride injection

[0123] Negative control: 0.9% sodium chloride injection

[0124] Positive control: distilled water

[0125] Contact route: tail vein injection

[0126] Test method: the test sample was immersed in 0.9% sodium chloride injection at a certain ratio and then incubated at 37° C. for 30 min in a water bath, and 0.2 ml of fresh anticoagulant rabbit blood was added, and the mixture was incubated at 37° C. for 60 min. Centrifugation was performed at 2500 rpm for 5 min, then the supernatant was taken, and the absorbance was measured at 545 nm by using an ultraviolet spectrophotometer to calculate the hemolysis rate.

[0127] Result: the hemolysis rate of the test sample is 0.2%.

[0128] Conclusion: according to GB/T 16886.4-2003, the hemolysis test results of the test samples meet the requirements on medical materials.

Embodiment 29

[0129] Stability and Integrity Test:

[0130] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; part of the solution B was taken and added with a small interfering nucleic acid drug (the same as embodiment 1, 10 ug/ml) to prepare solution C, the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, and drying was performed to form a film; then the solution B was instantaneously coated at high pressure and washing was performed by using water for injection; then the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for four times; then the solution A was instantaneously coated at high pressure and washing was performed by using water for injection; then the solution C was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the solution A and the solution C were alternatively coated in this way repeatedly for seven times, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the small interfering nucleic acid drug.

[0131] The multilayer film prepared above was placed in 1M NaCl solution and was incubated for a certain period of time, and the resulting slow-release solution was firstly subjected to filtration treatment through an ultrafiltration centrifuge tube with a molecular weight cut-off of 30 KDa to remove macromolecular substances. Then, it was filtered through an ultrafiltration centrifuge tube with a molecular weight cut-off of 3 KDa to remove salt ions in the slow-release solution, i.e., desalting treatment was performed. Finally, the treated slow-release solution was subjected to gel-running treatment as sample solution in an electrophoresis tank. Polyacrylamide gel electrophoresis (PAGE) was used herein to verify the gene integrity and related stability of siRNA released from the multilayer film.

[0132] The gel electrophoresis experiment in FIG. 1 verified the stability and integrity of the slow-release siRNA sequence. The siRNA-loaded tissue adhesive film was incubated in 1M NaCl solution, and the film was gradually ruptured and dissolved. After 6 days, it was difficult to observe the molded product. From the gel electrophoresis experiment in FIG. 1, it can be seen that the dissociated siRNA could not be observed after 6 days of desalting, while the dissociated siRNA could be clearly seen after 10 days of desalting. Accordingly, it can be inferred that, in the process of gradual rupture and dissolution, the lysate is a combination of siRNA and anionic and cationic groups. With degrading of the anionic and cationic groups, siRNA is gradually dissociated and maintains the stability and integrity of the siRNA sequence.

Embodiment 30

[0133] Cell Transfection Experiment:

[0134] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; part of the solution B was taken and added with a small interfering nucleic acid drug (the same as embodiment 1, 10 ug/ml) to prepare solution C, the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, and drying was performed to form a film; then the solution B was instantaneously coated at high pressure and washing was performed by using water for injection; then the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for four times; then the solution A was instantaneously coated at high pressure and washing was performed by using water for injection; then the solution C was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the solution A and the solution C were alternatively coated in this way repeatedly for two times, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the small interfering nucleic acid drug.

[0135] The tissue adhesive film prepared above was used as an experimental group, i.e., the experimental group was a film loading eGFP-siRNA which experienced alternate polyelectrolyte reaction twice, and the negative control group was a film loading common siRNA which experienced alternate polyelectrolyte reaction twice (the preparation method was the same as that of the experimental group, and only eGFP-siRNA was replaced with common siRNA). The experimental group and the negative control group were placed in a 6-well plate, respectively, and eGFP-HEK 293T cells were respectively inoculated into the 6-well plate (the cells were directly inoculated into wells in which no film was placed and were used as the blank control group). The changes in fluorescence intensity were observed on day 1, day 2 and day 3, and the results are shown in FIG. 2. In FIG. 3, the relative fluorescence intensities of the experimental group, the negative control group and the blank group are listed. It can be seen that the fluorescence intensity of the blank group and the negative control group is almost unchanged within 3 days, while the fluorescence intensity of the experimental group is significantly decreased with the increase of time within 3 days. Accordingly, it is verified that the tissue adhesive film loading eGFP siRNA can enable the fluorescence intensity of eGFP-HEK 293T cells to be reduced, and it indicates that eGFP siRNA has been transfected into the cells and has induced target gene silencing.

Embodiment 31

[0136] Stability and Integrity Test:

[0137] In an aseptic bench, a culture dish with diameter of 12 cm was provided, a high-molecular material film with the same diameter was put therein (subjected to washing, sterilization and depyrogenation treatment), washing was performed by using water for injection and drying was performed; solution A of a material with a cationic group (1 mg/ml collagen, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=3.5) and solution B of a material with an anionic group (1 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution A was added with an Afatinib cationic PEG nanometer microsphere (the concentration is 0.5 mg/ml), and uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: arginine-glycine-aspartic acid; the solution was respectively filled into a high-pressure instantaneous coating device, firstly the solution A was instantaneously coated at high pressure into the culture dish, drying was performed, then the solution B was instantaneously coated at high pressure, washing was performed by using water for injection, the solution A and the solution B were alternately coated in this way repeatedly for 200 times; and finally the solution A was instantaneously coated at high pressure, washing was performed by using water for injection, drying was performed, the film was peeled off, an adhesive side of the film was washed by using water for injection, and dried to obtain an implantable tissue adhesive film loading the Afatinib drug.

[0138] The multilayer film prepared above was placed in 1M NaCl solution and was incubated for a certain period of time, the obtained slow-release solution was purified, Afatinib could be obtained through separation, and the amount of Afatinib obtained through separation was not significantly reduced relative to the amount of Afatinib added during preparation. Accordingly, it can be seen that the stability and integrity of the drug are maintained.

Embodiment 32

[0139] Cell Transfection Experiment:

[0140] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with a vascular endothelial growth factor (VEGF) (0.5 mg/ml), and uniform stirring with a targeting polypeptide (1*10.sup.−4 mg/ml) was performed, wherein an amino acid sequence of the targeting polypeptide was: valine-glycine-valine-alanine-proline-glycine; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 80 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain a tissue adhesive film loading the vascular endothelial growth factor (VEGF) drug.

[0141] The multilayer film prepared above was placed in 1M NaCl solution and was incubated for a certain period of time, the obtained slow-release solution was purified, the vascular endothelial growth factor could be obtained through separation, and as detected by using a VEGF detection kit, the amount of the VEGF obtained through separation was not significantly reduced relative to the amount of the VEGF added during preparation. Accordingly, it can be seen that the stability and integrity of the drug are maintained.

Embodiment 33

[0142] In an aseptic bench, a high-molecular material plate was provided (subjected to washing, sterilization and depyrogenation treatment), and drying was performed; solution A of a material with a cationic group (2 mg/ml chitosan, 0.1 mol/L acetic acid, 0.2 mol/L NaCl, pH=4) and solution B of a material with an anionic group (2 mg/ml hyaluronic acid, 0.15 mol/L NaCl, pH=6) were respectively prepared; the solution B was added with Polyporus polysaccharide (0.5 mg/ml), and uniform stirring was performed; the plate was firstly immersed in the solution A for 20 min, taken out, put in washing solution for washing and dried; and then the plate was immersed in the solution B for 30 min, taken out, put in washing solution for washing and dried, the operations were alternately performed for 100 times in this way, finally washing was performed by using water for injection, drying was performed and the film was peeled off to obtain a tissue adhesive film loading the Polyporus polysaccharide drug.

[0143] The multilayer film prepared above was placed in 1M NaCl solution and was incubated for a certain period of time to obtain slow-release solution. By measuring the polysaccharide components in the slow-release solution, it can be found that the amount of the polysaccharide in the slow-release solution obtained through separation was not significantly reduced relative to the amount of the polysaccharide added during preparation. Accordingly, it can be seen that the stability and integrity of the drug are maintained.

[0144] To sum up, the present invention effectively overcomes various disadvantages in the prior art and thus has a great industrial utilization value.

[0145] The above-mentioned embodiments are just used for exemplarily describing the principle and effect of the present invention instead of limiting the present invention. One skilled in the art may make modifications or changes to the above-mentioned embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or changes made by those who have common knowledge in the art without departing from the spirit and technical thought disclosed by the present invention shall be still covered by the claims of the present invention.