High-permeability drug coating balloon as well as preparation method and application thereof

Abstract

The invention discloses a high-permeability drug coating balloon as well as a preparation method and application thereof, and belongs to the technical field of cardiovascular disease treatment materials. The preparation method comprises the following steps: pretreating a sponge spicule, sequentially placing the pretreated sponge spicule in a sodium hydroxide solution and a hydrochloric acid solution for ultrasonic treatment, washing and drying; preparing a silane coupling agent hydrolysate; preparing sponge spicule dispersion liquid; heating the sponge spicule dispersion liquid, adding the silane coupling agent hydrolysate, and carrying out a stirring reaction to obtain a surface-modified sponge spicule; mixing a drug with the surface-modified sponge spicule, and adding absolute ethyl alcohol for dispersing and grafting to obtain a drug-loaded sponge spicule; and dissolving the drug-loaded sponge spicule in a shellac ethanol solution for spin coating, and drying to obtain the drug-loaded sponge spicule. The invention further discloses the high-permeability drug coating balloon prepared by the method and application of the high-permeability drug coating balloon. The high-permeability drug coating balloon has the advantages of low drug dosage, high drug administration efficiency, low toxic and side effects and the like, and has a wide application prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of cardiovascular disease treatment materials, and specifically relates to a high-permeability drug-coated balloon and a preparation method and application thereof. Background Art

[0002] Cardiovascular diseases seriously endanger people's health and safety. Among them, atherosclerosis is one of the important driving factors of cardiovascular diseases. Prevention and treatment of atherosclerosis is an important means to reduce the incidence of cardiovascular diseases. Drug-coated balloon (DCB) is an "intervention without implantation" method for treating atherosclerosis, which can effectively avoid the risk of restenosis caused by material implantation. However, due to factors such as blood washout and drug residue, the drug utilization rate of DCB is not very high.

[0003] Sponges are an ancient porous filter-feeding animal on Earth. The bone needles formed by biomineralization in their bodies are called sponge spicules. The spicules and the skeleton together constitute the skeleton of sponges and play the role of supporting sponges. Studies have found that sponge spicules have good mechanical properties, high mechanical strength, and the sharp structure at the end makes them function like microneedles. They can stay in the stratum corneum of the skin for a long time to form a large number of microchannels, effectively promoting the absorption of drugs or active ingredients. At present, research on them mainly focuses on their optical properties, nanostructures, bionics, and pharmaceutical transdermal agents.

[0004] As a unique biomaterial, the chemical properties and physical structure of sponge bone needles are incompatible with the drugs and coating materials used in drug-coated balloons. This incompatibility can lead to uneven drug release, coating shedding or performance degradation. How to prepare a coated balloon with high drug utilization and high permeability is a necessary method to solve the problems currently existing in drug-coated balloon treatment. Summary of the invention

[0005] The problem to be solved by the present invention is to provide a high-permeability drug-coated balloon and a preparation method and application thereof, so as to give the drug-coated balloon the effects of low drug dosage, high-efficiency drug administration and low toxic side effects, and solve the problems of low drug utilization rate and high drug loss rate.

[0006] The technical solution adopted to solve the technical problem is to provide a method for preparing a high-permeability drug-coated balloon, comprising the following steps:

[0007] (1) After pretreatment, the sponge spicules are placed in a sodium hydroxide solution and a hydrochloric acid solution in sequence for ultrasonication, washing and drying to obtain sponge spicules with surface silanol activated;

[0008] (2) mixing a silane coupling agent, water and anhydrous ethanol to prepare a silane coupling agent hydrolyzate;

[0009] (3) mixing the sponge spicules activated with silanol groups on the surface with anhydrous ethanol and performing ultrasonication to obtain a sponge spicule dispersion;

[0010] (4) heating the sponge spicule dispersion and adding the silane coupling agent hydrolyzate to carry out stirring reaction, washing and drying to obtain the surface-modified sponge spicule;

[0011] (5) mixing the drug with the surface-modified sponge spicules and adding anhydrous ethanol for dispersion and grafting to obtain drug-loaded sponge spicules;

[0012] (6) The drug-loaded sponge bone needle is dissolved in a shellac ethanol solution and spin-coated, and after drying, a highly permeable drug-coated balloon is obtained.

[0013] The beneficial effects of the above technical solution adopted in the present invention are as follows: in the present invention, the pretreated sponge spicules are treated with sodium hydroxide and hydrochloric acid in sequence, the sodium hydroxide treatment can remove oil and impurities on the surface of the sponge spicules, and then the hydrochloric acid etching is used to further clean and increase the roughness of the surface of the spicules, while introducing or enhancing the presence of silanol (Si-OH) to provide active sites for subsequent chemical reactions; the sponge spicules with surface silanol activated are evenly dispersed in anhydrous ethanol, which helps to reduce the agglomeration of the sponge spicules and improve the efficiency of subsequent reactions; by heating the sponge spicule dispersion A silane coupling agent is added to promote the condensation reaction between the hydrolysis product of the silane coupling agent and the silanol group on the surface of the sponge spicule, thereby firmly introducing the corresponding functional groups. The effective loading of the drug on the surface of the sponge spicule is achieved through the interaction between the drug group and the corresponding functional group, thereby improving the stability and targeting of the drug. Finally, it is dissolved in a shellac ethanol solution. Shellac has good film-forming properties and biocompatibility. As a coating material, it can protect the drug-loaded sponge spicules and control the release rate of the drug. The high-permeability drug-coated balloon finally obtained is uniform and smooth, and has good sealing and drug permeability.

[0014] Preferably, step (1) comprises the following steps: grinding the sponge spicule for 2 to 4 minutes and centrifuging at 3500 to 4500 r / min, placing it in a sodium hydroxide solution with a molar concentration of 1 to 4 mol / L and ultrasonicating it for 30 to 40 minutes, washing and drying it, and then placing it in a hydrochloric acid solution with a volume concentration of 4 to 6% and ultrasonicating it for 30 to 40 minutes, washing and drying it to obtain a sponge spicule with surface silanol activated.

[0015] The beneficial effects of the above-mentioned technical scheme adopted in the present invention are as follows: if the sponge spicules are too long, a large number of them will lie flat on the coating and cannot reach a vertical state; if they are too short, they cannot penetrate the vascular endothelium, thus affecting the treatment effect; the size of the sponge spicules can be further broken by grinding, and the sponge spicules of different sizes can be separated by the action of centrifugal force, so as to obtain sponge spicules of tiny size and needle-like structure.

[0016] More preferably, step (1) includes the following steps: grinding the sponge bone needle for 3 minutes and centrifuging at 4000 r / min, placing it in a sodium hydroxide solution with a molar concentration of 1 mol / L and ultrasonicating it for 40 minutes, washing and drying it, and then placing it in a hydrochloric acid solution with a volume concentration of 5% and ultrasonicating it for 40 minutes, washing and drying it to obtain a sponge bone needle with surface silanol activated.

[0017] Preferably, in step (2), the silane coupling agent is (3-aminopropyl)triethoxysilane; and the volume ratio of the silane coupling agent, water and anhydrous ethanol is 1:1:5-7.

[0018] More preferably, in step (2), the silane coupling agent is (3-aminopropyl)triethoxysilane; and the volume ratio of the silane coupling agent, water and anhydrous ethanol is 1:1:6.

[0019] Preferably, in step (3), the material-liquid ratio of the sponge spicules with surface silanol activated to anhydrous ethanol is 0.1 g:10-15 mL; and the ultrasonic time is 10-20 min.

[0020] More preferably, in step (3), the material-liquid ratio of the sponge spicules with surface silanol activated to anhydrous ethanol is 0.1 g:13 mL; and the ultrasonic time is 15 min.

[0021] Preferably, the heating temperature in step (4) is 85-95°C; the volume ratio of the sponge spicule dispersion to the silane coupling agent hydrolyzate is 10-15:5-10; the stirring reaction temperature is 55-65°C, and the time is 5-7h.

[0022] More preferably, in step (4), the heating temperature is 90° C.; the volume ratio of the sponge spicule dispersion to the silane coupling agent hydrolyzate is 13:8; the stirring reaction temperature is 60° C., and the reaction time is 6 h.

[0023] Preferably, in step (5), the mass ratio of the drug to the surface-modified sponge spicule is 0.1:2-3; the dispersion grafting temperature is 25-30° C., and the time is 10-15 min.

[0024] More preferably, in step (5), the mass ratio of the drug to the surface-modified sponge spicule is 0.1:2.8; the dispersion grafting temperature is 30° C., and the time is 10 min.

[0025] More preferably, the drug is colchicine, pravastatin or paclitaxel; and the drug grafting rate on the drug-loaded sponge bone needle is 6-7%.

[0026] The beneficial effect of the above technical solution of the present invention is that when the drug is colchicine, the amide in the colchicine can be combined with the amino group on the modified sponge spicule to achieve dispersed grafting of colchicine. The specific process is as follows:

[0027]

[0028] More preferably, the drug grafting rate on the drug-loaded sponge spicules is 6.955%.

[0029] Preferably, step (6) comprises the following steps: dissolving the drug-loaded sponge spicules in a shellac ethanol solution having a mass concentration of 90 to 110 mg / mL, so that the mass concentration of the drug-loaded sponge spicules is 9 to 10 mg / mL, to obtain a mixed solution; spin coating the mixed solution at 300 to 500 r / min for 10 to 15 s, and then spin coating at 1500 to 2500 r / min for 25 to 35 s using a homogenizer, and obtaining a highly permeable drug-coated balloon after drying.

[0030] More preferably, step (6) comprises the following steps: dissolving the drug-loaded sponge spicules in a shellac ethanol solution having a mass concentration of 100 mg / mL to obtain a mixed solution having a mass concentration of 9.5 mg / mL; spin coating the mixed solution at 350 r / min for 12 s, and then at 2000 r / min for 30 s using a homogenizer, and obtaining a highly permeable drug-coated balloon after drying.

[0031] More preferably, the mass of the drug-loaded sponge spicule on the high permeability drug-coated balloon is 3 μg / mm 2 .

[0032] More preferably, the thickness of the high permeability drug-coated balloon is 1 to 3 μm.

[0033] The present invention also provides a high-permeability drug-coated balloon prepared by the preparation method.

[0034] The present invention also provides the use of the high-permeability drug-coated balloon in preparing materials for treating atherosclerosis.

[0035] The present invention has the following beneficial effects:

[0036] The high permeability drug-coated balloon of the present invention uses shellac as an excipient, which helps the drug-loaded bone needles adhere to the balloon after being mixed with the sponge spicules. When the drug-coated balloon reaches the injury site, the sponge spicules with good mechanical properties can penetrate the vascular endothelium to form micro-channels, promote drug absorption, and improve drug utilization. The high permeability drug-coated balloon of the present invention can increase drug utilization by nearly 1.8 times, has the advantages of low drug dosage, efficient drug administration, and low toxic side effects, solves the problems of low drug utilization and drug loss, and has broad application prospects and practical value. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 This is a transmission electron microscope image of a bee sponge spicule;

[0038] Figure 2 The figures are 40 times optical microscope images of bee sponge spicules after different pretreatments; wherein (a) is the bee sponge spicules not pretreated in Comparative Example 4; (b) is the bee sponge spicules pretreated in Comparative Example 1; (c) is the bee sponge spicules pretreated in Comparative Example 2; (d) is the bee sponge spicules pretreated in Example 1;

[0039] Figure 3 This is a 100x optical microscope image of the pretreated bee sponge spicules in Example 1;

[0040] Figure 4 3D images of the highly permeable drug-coated balloon prepared in Example 1; (a) is an Imaris3D reconstructed image after confocal layer scanning; (b) is an Imaris coating thickness construction result image;

[0041] Figure 5 The images are reconstructed by Imaris; (a) is the 3D confocal layer scanning result of the hydrogel after administration of the coating containing bone needles; (b) is the 3D confocal layer scanning result of the hydrogel after administration of the coating without bone needles;

[0042] Figure 6 The graphs are the results of the in vitro drug release experiment of the drug-loaded sponge bone needles; among them, (a) is the standard curve of colchicine; (b) is the drug release curve of the drug-loaded sponge bone needles. DETAILED DESCRIPTION

[0043] In order to make the purpose, technical scheme and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention, that is, the embodiments described are only part of the embodiments of the present invention, rather than all of the embodiments.

[0044] Therefore, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present invention.

[0045] The features and properties of the present invention are further described in detail below in conjunction with the examples. The sponge spicules are bee sponge spicules provided by Xi'an Mixianer Biotechnology Co., Ltd.; the shellac is provided by Shanghai MacLean Biochemical Technology Co., Ltd.; and the colchicine is provided by Shanghai Yien Chemical Technology Co., Ltd.

[0046] Example 1

[0047] A method for preparing a highly permeable drug-coated balloon comprises the following steps:

[0048] (1) grinding the bee sponge spicule for 3 minutes and centrifuging it at 4000 r / min, then placing it in a sodium hydroxide solution with a molar concentration of 1 mol / L for ultrasonic treatment for 40 minutes, centrifugally washing it twice with deionized water and once with anhydrous ethanol, and then drying it in an oven at 37° C., then placing it in a hydrochloric acid solution with a volume concentration of 5% for ultrasonic treatment for 40 minutes, centrifugally washing it twice with deionized water and once with anhydrous ethanol, and then drying it in an oven at 37° C., to obtain sponge spicules with surface silanol activated;

[0049] (2) preparing a silane coupling agent hydrolyzate by mixing (3-aminopropyl)triethoxysilane, RO water and anhydrous ethanol in a volume ratio of 1:1:6;

[0050] (3) 0.1 g of the sponge spicules activated with surface silanol groups was mixed with 13 mL of anhydrous ethanol and ultrasonicated for 15 min to obtain a sponge spicule dispersion;

[0051] (4) heating the sponge spicule dispersion in a water bath under stirring, and when the temperature stabilizes at 90° C., dropping 8 mL of silane coupling agent hydrolyzate, and then stirring and reacting at 60° C. for 6 h, repeatedly centrifugally washing with anhydrous ethanol 3 times, centrifugally washing with RO water once, and then drying in an oven at 37° C. to obtain sponge spicules with surface amino groups modified;

[0052] (5) 0.005 g of colchicine was mixed with 0.14 g of surface amino-modified sponge spicules and 2 mL of anhydrous ethanol was added to perform dispersion grafting at 1000 r / min at 30° C. for 10 min, and then stirred at room temperature until the ethanol evaporated to obtain drug-loaded sponge spicules;

[0053] (6) Weigh 100 mg of shellac and dissolve it in 1 mL of anhydrous ethanol to obtain a shellac ethanol solution with a mass concentration of 100 mg / mL; dissolve the drug-loaded sponge spicules in the shellac ethanol solution with a mass concentration of 100 mg / mL to make the drug-loaded sponge spicules have a mass concentration of 9.5 mg / mL to obtain a mixed solution; place the PU disc in the suction cup of a glue spreader, turn on the vacuum pump to tightly fix the PU disc on the suction cup, use a pipette to draw 100 μL of the mixed solution and drop it into the center of the disc, turn on the glue spreader, spin coat at 350 r / min for 12 s, and then spin coat at 2000 r / min for 30 s, and place in an oven at 37°C to dry to obtain a highly permeable drug-coated balloon.

[0054] This embodiment also provides a high permeability drug-coated balloon prepared by the above preparation method. The drug grafting rate on the drug-loaded sponge spicules in the high permeability drug-coated balloon is 6.955%, and the mass of the drug-loaded sponge spicules on the high permeability drug-coated balloon is 3 μg / mm 2 .

[0055] Example 2

[0056] A method for preparing a highly permeable drug-coated balloon comprises the following steps:

[0057] (1) grinding the bee sponge spicule for 2 minutes and centrifuging at 4500 r / min, placing it in a sodium hydroxide solution with a molar concentration of 4 mol / L for ultrasonic treatment for 30 minutes, centrifugally washing it twice with deionized water and once with anhydrous ethanol, and then drying it in an oven at 37° C., and then placing it in a hydrochloric acid solution with a volume concentration of 6% for ultrasonic treatment for 30 minutes, centrifugally washing it twice with deionized water and once with anhydrous ethanol, and then drying it in an oven at 37° C., to obtain sponge spicules with surface silanol activated;

[0058] (2) preparing a silane coupling agent hydrolyzate by mixing (3-aminopropyl)triethoxysilane, RO water and anhydrous ethanol in a volume ratio of 1:1:5;

[0059] (3) 0.1 g of the sponge spicules activated with surface silanol groups was mixed with 10 mL of anhydrous ethanol and ultrasonicated for 10 min to obtain a sponge spicule dispersion;

[0060] (4) heating the sponge spicule dispersion in a water bath under stirring, and when the temperature stabilizes at 85° C., dropping 5 mL of silane coupling agent hydrolyzate, and then stirring and reacting at 55° C. for 7 h, repeatedly centrifugally washing with anhydrous ethanol 3 times, centrifugally washing with RO water once, and then drying in an oven at 37° C. to obtain sponge spicules with surface amino groups modified;

[0061] (5) 0.005 g of colchicine was mixed with 0.10 g of surface amino-modified sponge spicules and 1 mL of anhydrous ethanol was added to perform dispersion grafting at 1000 r / min at 30° C. for 15 min and then stirred at room temperature until the ethanol evaporated to obtain drug-loaded sponge spicules;

[0062] (6) Weigh 90 mg of shellac and dissolve it in 1 mL of anhydrous ethanol to obtain a shellac ethanol solution with a mass concentration of 90 mg / mL; dissolve the drug-loaded sponge spicules in the shellac ethanol solution with a mass concentration of 90 mg / mL to make the drug-loaded sponge spicules have a mass concentration of 9.5 mg / mL to obtain a mixed solution; place the PU disc in the suction cup of a glue spreader, turn on the vacuum pump to tightly fix the PU disc on the suction cup, use a pipette to draw 100 μL of the mixed solution and add it dropwise to the center of the disc, turn on the glue spreader, spin coat at 300 r / min for 15 s, and then spin coat at 1500 r / min for 35 s, and place in an oven at 37°C to dry to obtain a highly permeable drug-coated balloon.

[0063] Example 3

[0064] A method for preparing a highly permeable drug-coated balloon comprises the following steps:

[0065] (1) grinding the bee sponge spicules for 4 minutes and centrifuging them at 3500 r / min, then placing them in a sodium hydroxide solution with a molar concentration of 2 mol / L for ultrasonic treatment for 40 minutes, centrifugally washing them twice with deionized water and once with anhydrous ethanol, and then drying them in an oven at 37° C., then placing them in a hydrochloric acid solution with a volume concentration of 4% for ultrasonic treatment for 30 minutes, centrifugally washing them twice with deionized water and once with anhydrous ethanol, and then drying them in an oven at 37° C., to obtain sponge spicules with surface silanol activated;

[0066] (2) preparing a silane coupling agent hydrolyzate by mixing (3-aminopropyl)triethoxysilane, RO water and anhydrous ethanol in a volume ratio of 1:1:7;

[0067] (3) 0.1 g of the sponge spicules activated with surface silanol groups was mixed with 15 mL of anhydrous ethanol and ultrasonicated for 20 min to obtain a sponge spicule dispersion;

[0068] (4) heating the sponge spicule dispersion in a water bath under stirring, and when the temperature stabilizes at 95° C., dropping 10 mL of silane coupling agent hydrolyzate, and then stirring and reacting at 65° C. for 5 h, repeatedly centrifugally washing with anhydrous ethanol 3 times, centrifugally washing with RO water once, and then drying in an oven at 37° C. to obtain sponge spicules with surface amino groups modified;

[0069] (5) 0.005 g of colchicine was mixed with 0.15 g of surface amino-modified sponge spicules and 2 mL of anhydrous ethanol was added to perform dispersion grafting at 1000 r / min at 30° C. for 10 min, and then stirred at room temperature until the ethanol evaporated to obtain drug-loaded sponge spicules;

[0070] (6) Weigh 110 mg of shellac and dissolve it in 1 mL of anhydrous ethanol to obtain a shellac ethanol solution with a mass concentration of 110 mg / mL; dissolve the drug-loaded sponge spicules in the shellac ethanol solution with a mass concentration of 110 mg / mL to make the drug-loaded sponge spicules have a mass concentration of 9.5 mg / mL to obtain a mixed solution; place the PU disc in the suction cup of a glue spreader, turn on the vacuum pump to tightly fix the PU disc on the suction cup, use a pipette to draw 100 μL of the mixed solution and add it dropwise to the center of the disc, turn on the glue spreader, spin coat at 500 r / min for 10 s, then spin coat at 2500 r / min for 25 s, and place in an oven at 37°C to dry to obtain a highly permeable drug-coated balloon.

[0071] Comparative Example 1

[0072] A method for preparing a highly permeable drug-coated balloon, compared with Example 1, the difference is that: step (1) is: grinding a bee sponge spicule for 3 minutes and centrifuging it at 1000r / min, placing it in a sodium hydroxide solution with a molar concentration of 1mol / L for ultrasonic treatment for 40 minutes, centrifugally washing it with deionized water twice, centrifugally washing it with anhydrous ethanol once, and then drying it in an oven at 37°C, then placing it in a hydrochloric acid solution with a volume concentration of 5% for ultrasonic treatment for 40 minutes, centrifugally washing it with deionized water twice, centrifugally washing it with anhydrous ethanol once, and then drying it in an oven at 37°C to obtain a sponge spicule with surface silanol activated; the remaining steps are the same as Example 1.

[0073] Comparative Example 2

[0074] A method for preparing a highly permeable drug-coated balloon, compared with Example 1, differs in that: step (1) comprises: grinding a bee sponge spicule for 3 minutes and subjecting it to a centrifugal treatment at 2000 r / min, placing it in a sodium hydroxide solution with a molar concentration of 1 mol / L for ultrasonic treatment for 40 minutes, washing it by centrifugation twice with deionized water and once with anhydrous ethanol, and then drying it in an oven at 37°C, and then placing it in a hydrochloric acid solution with a volume concentration of 5% for ultrasonic treatment for 40 minutes, and then centrifugation washing it by centrifugation twice with deionized water and once with anhydrous ethanol, and then drying it in an oven at 37°C, to obtain a sponge spicule with surface silanol activated; the remaining steps are the same as those in Example 1.

[0075] Comparative Example 3

[0076] A method for preparing a highly permeable drug-coated balloon, compared with Example 1, the difference is that: step (1) is: after grinding the bee sponge spicule for 3 minutes, placing it in a sodium hydroxide solution with a molar concentration of 1 mol / L for ultrasonication for 40 minutes, centrifugally washing it with deionized water twice, centrifugally washing it with anhydrous ethanol once, and then drying it in an oven at 37°C, then placing it in a hydrochloric acid solution with a volume concentration of 5% for ultrasonication for 40 minutes, centrifugally washing it with deionized water twice, centrifugally washing it with anhydrous ethanol once, and then drying it in an oven at 37°C to obtain a sponge spicule with surface silanol activated; the remaining steps are the same as Example 1.

[0077] Comparative Example 4

[0078] A method for preparing a highly permeable drug-coated balloon, compared with Example 1, the difference is that: step (1) is: placing a bee sponge spicule in a sodium hydroxide solution with a molar concentration of 1 mol / L for ultrasonication for 40 minutes, centrifugally washing it with deionized water twice, centrifugally washing it with anhydrous ethanol once, and then drying it in an oven at 37°C, and then placing it in a hydrochloric acid solution with a volume concentration of 5% for ultrasonication for 40 minutes, centrifugally washing it with deionized water twice, centrifugally washing it with anhydrous ethanol once, and then drying it in an oven at 37°C to obtain a sponge spicule with surface silanol activated; the remaining steps are the same as Example 1.

[0079] Comparative Example 5

[0080] A method for preparing a drug-coated balloon comprises the following steps:

[0081] 100 mg of shellac was weighed and dissolved in 1 mL of anhydrous ethanol to obtain a shellac ethanol solution with a mass concentration of 100 mg / mL; 0.005 g of colchicine was dissolved in the shellac ethanol solution with a mass concentration of 100 mg / mL to make the mass concentration of colchicine 9.5 mg / mL to obtain a mixed solution; a PU disc was placed in the suction cup of a glue spreader, the vacuum pump was turned on to tightly fix the PU disc on the suction cup, 100 μL of the mixed solution was drawn with a pipette and dropped into the center of the disc, the glue spreader was turned on, spin-coated at 350 r / min for 12 s, and then spin-coated at 2000 r / min for 30 s, and dried in an oven at 37°C to obtain a drug-coated balloon.

[0082] Experimental example

[0083] 1. Use transmission electron microscopy to analyze the morphology of bee sponge spicules, such as Figure 1 As shown, the bee sponge spicule is a uniaxial spicule with sharp needle-like structures at both ends.

[0084] 2. The bee sponge spicules of Example 1, Comparative Examples 1-2 after pretreatment (grinding bee sponge spicules and then centrifuging them), and Comparative Example 4 without pretreatment were observed and analyzed using an optical microscope, and the length of the spicules was measured using Image J. The results are as follows: Figures 2-3 As shown. Figure 2 It can be seen from the 40x optical microscope shooting results that the size of the sponge bone needle raw material (Comparative Example 4) is about 1 to 200 μm, and a small number of bone needles are 1 to 5 μm in length; the size of the ground bone needles (Comparative Example 1) centrifuged at 1000 r / min is 1 to 80 μm, and the distribution density of the bone needles containing the needle tip structure after breaking is low; the size of the ground bone needles (Comparative Example 2) centrifuged at 2000 r / min is 1 to 50 μm, and more than half of them are bone needles without the needle tip structure after breaking; the size of the ground bone needles (Example 1) centrifuged at 4000 r / min is 1 to 10 μm, and most of them are bone needles with a needle tip structure after breaking; the ground bone needles (Example 1) centrifuged at 4000 r / min were further observed using a 100x optical microscope, as shown Figure 3 As shown in the figure, most of the ground bone needles centrifuged at 4000r / min present a tiny needle-like structure, and the size of the tiny needle-like bone needles after breaking is about 1 to 5μm. The analysis results show that if the sponge bone needles are too long, a large number of them will lie flat on the coating and cannot reach a vertical state; if they are too short, they cannot penetrate the vascular endothelium, thus affecting the treatment effect; after grinding the bone needles for 3 minutes to break the size, centrifuging at a speed of 4000r / min, a tiny sponge bone needle with a needle-like structure can be obtained, which is conducive to subsequent use.

[0085] 3. The highly permeable drug-coated balloon prepared in Example 1 was confocal scanned, and the 3D layer scanning results were reconstructed using the Imaris software. The results are as follows: Figure 4 As shown. Figure 4 As can be seen in Figure (a), the surface of the high permeability drug-coated balloon prepared by the present invention is relatively uniform, the drug-loaded sponge spicules are evenly distributed in the coating, and some of the drug-loaded sponge spicules are in a vertical state in the coating; Figure 4 As shown in (b), Imaris constructed a side image for characterizing the coating thickness; Image J was used to measure the coating thickness, and the results showed that the average thickness of the high-permeability drug-coated balloon was 2.778 μm.

[0086] 4. Fluorescent carbon dots were selected to replace the colchicine in Example 1 and Comparative Example 5, and 15% GelMa hydrogel was simulated for drug administration and drug penetration analysis. After confocal layer scanning, 3D reconstruction was performed using Imaris. The average fluorescence intensity of the fluorescent carbon dots that penetrated into the hydrogel was analyzed by Image J. It was found that the fluorescence intensity of the hydrogel administered with the coating without spicules (Comparative Example 5) was 140.322, and the average fluorescence intensity of the gel after administration with the coating containing spicules (Example 1) was 243.586, which was about 1.73 times that of the drug without the coating containing spicules. The results are as follows Figure 5 As shown, the sponge spicules in the coating can significantly promote drug penetration.

[0087] 5. In order to explore whether the drug release amount of the prepared drug-loaded sponge bone needle (COL-SHS) at different times meets the concentration requirements for the treatment of atherosclerosis, an in vitro drug release experiment was conducted on COL-SHS: 30 mg of COL-SHS was weighed and placed in 20 mL of 10% PBS buffer solution, and the drug was released by shaking at 37°C. At different times (15min, 0.5h, 1h, 2h, 4h, 6h, 12h, 24h, 30h, 36h, 48h), 6 mL of the drug release solution was taken and divided into 2 centrifuge tubes, and then 6 mL of PBS buffer solution was added to the drug release system. The absorbance of the drug release solution obtained at different times was measured using an ultraviolet spectrophotometer.

[0088] Since the solubility of COL in ethanol and PBS solutions is quite different, before exploring the content of COL in PBS solution, it is necessary to find out the relationship between the concentration of COL in PBS solvent and absorbance: prepare a colchicine standard solution with 10% PBS as solvent, and measure the absorbance at 350nm to obtain the relationship between the colchicine solution with PBS as solvent and absorbance. The results are as follows: Figure 6 (a) As shown. Based on the colchicine solution standard curve with PBS as the solvent, the concentration of COL in the sample solution at different times can be measured, and the drug release rate at each time can be calculated according to the following cumulative release rate formula:

[0089]

[0090] In the above formula, E r is the cumulative release rate of COL, %; V e is the PBS replacement volume, 6 mL; V0 is the total volume of drug release, 20 mL; C i is the drug concentration of the release solution at the i-th replacement sampling; m is the mass of the drug contained in the drug-loaded bone needle; n is the number of times PBS is replaced.

[0091] The in vitro drug release curve of COL-SHS was drawn with the drug release time as the horizontal axis and the measured cumulative release rate and drug release amount as the double vertical axes. The results are shown in Figure 6 (b) as shown.

[0092] The cumulative drug release results showed that the drug release of COL-SHS in PBS increased slowly with time and tended to be stable after 30 hours. The maximum drug release was about 1358μg, and the cumulative drug release rate was up to about 60%, indicating a good drug release effect.

[0093] The present invention has been described according to the above embodiments. It should be understood that the above embodiments do not limit the present invention in any form. Any technical solutions obtained by equivalent replacement or equivalent transformation shall fall within the scope of the present invention.

Claims

1. A method for preparing a highly permeable drug-coated balloon, characterized in that: The following steps are involved: (1) After pretreatment, the sponge spicules are placed in a sodium hydroxide solution and a hydrochloric acid solution in sequence for ultrasonication, washing and drying to obtain sponge spicules with surface silanol activated; (2) mixing a silane coupling agent, water and anhydrous ethanol to prepare a silane coupling agent hydrolyzate; (3) mixing the sponge spicules activated with silanol groups on the surface with anhydrous ethanol and performing ultrasonication to obtain a sponge spicule dispersion; (4) heating the sponge spicule dispersion and adding the silane coupling agent hydrolyzate to carry out stirring reaction, washing and drying to obtain the surface-modified sponge spicule; (5) mixing the drug with the surface-modified sponge spicules and adding anhydrous ethanol for dispersion and grafting to obtain drug-loaded sponge spicules; (6) The drug-loaded sponge bone needle is dissolved in a shellac ethanol solution and spin-coated, and after drying, a highly permeable drug-coated balloon is obtained.

2. The method for preparing a highly permeable drug-coated balloon according to claim 1, characterized in that: The step (1) comprises the following steps: grinding the sponge spicule for 2 to 4 minutes and centrifuging at 3500 to 4500 r / min, placing it in a sodium hydroxide solution with a molar concentration of 1 to 4 mol / L and ultrasonicating it for 30 to 40 minutes, washing and drying it, and then placing it in a hydrochloric acid solution with a volume concentration of 4 to 6% and ultrasonicating it for 30 to 40 minutes, washing and drying it to obtain a sponge spicule with surface silanol activated.

3. The method for preparing a highly permeable drug-coated balloon according to claim 1, characterized in that: The silane coupling agent in step (2) is (3-aminopropyl)triethoxysilane; the volume ratio of the silane coupling agent, water and anhydrous ethanol is 1:1:5-7.

4. The method for preparing a highly permeable drug-coated balloon according to claim 1, characterized in that: In the step (3), the liquid ratio of the sponge spicules with surface silanol activated to anhydrous ethanol is 0.1 g:10-15 mL; and the ultrasonic time is 10-20 min.

5. The method for preparing a highly permeable drug-coated balloon according to claim 1, characterized in that: The heating temperature in step (4) is 85-95° C.; the volume ratio of the sponge spicule dispersion to the silane coupling agent hydrolyzate is 10-15:5-10; the stirring reaction temperature is 55-65° C., and the reaction time is 5-7 hours.

6. The method for preparing a highly permeable drug-coated balloon according to claim 1, characterized in that: In the step (5), the mass ratio of the drug to the surface-modified sponge spicule is 0.1:2-3; the dispersion grafting temperature is 25-30° C., and the time is 10-15 min.

7. The method for preparing a highly permeable drug-coated balloon according to claim 1 or 6, characterized in that: The drug is colchicine, pravastatin or paclitaxel; and the drug grafting rate on the drug-loaded sponge bone needle is 6-7%.

8. The method for preparing a highly permeable drug-coated balloon according to claim 1, characterized in that: The step (6) comprises the following steps: dissolving the drug-loaded sponge spicules in a shellac ethanol solution with a mass concentration of 90 to 110 mg / mL, so that the mass concentration of the drug-loaded sponge spicules is 9 to 10 mg / mL, to obtain a mixed solution; spin coating the mixed solution at 300 to 500 r / min for 10 to 15 seconds, and then spin coating at 1500 to 2500 r / min for 25 to 35 seconds using a homogenizer, and obtaining a highly permeable drug-coated balloon after drying.

9. A highly permeable drug-coated balloon prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the high permeability drug-coated balloon according to claim 9 in preparing materials for treating atherosclerosis.

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