CN111001044A

CN111001044A - Medicine balloon, preparation of medicine coated on medicine balloon and preparation method of medicine balloon

Info

Publication number: CN111001044A
Application number: CN201911394205.1A
Authority: CN
Inventors: 胡帅领; 王森; 王鼎曦; 黄中嵘; 王海兰; 戴志豪
Original assignee: Shanghai Shenqi Medical Technology Co Ltd
Current assignee: Shanghai Shenqi Medical Technology Co Ltd
Priority date: 2019-12-30
Filing date: 2019-12-30
Publication date: 2020-04-14

Abstract

The invention belongs to the technical field of materials of catheters or coated catheters, and discloses a drug balloon, preparation of a coated drug of the drug balloon and a preparation method of the drug balloon. The preparation method of the drug balloon coated drug comprises the following steps: (1) preparing an organic phase solution of the drug and the polymer; (2) preparing an aqueous phase solution containing a surfactant and a stabilizer; (3) adding the organic phase solution into the aqueous phase solution, and emulsifying the aqueous phase solution; (4) removing the organic solvent; (5) and (4) centrifuging the nanoparticle suspension prepared in the step (4), discarding the supernatant of the centrifuged sample tube, adding deionized water for resuspension and centrifuging, and washing away free drugs and residual organic solvents. The nano particles of the technical scheme can be quickly absorbed by blood vessels, provide slow release performance and prolong the retention time of the medicine in tissues. Meanwhile, the cationic liposome coating can ensure that the nano particles are adhered to the surface of the vessel wall, and long-time drug release is ensured.

Description

Medicine balloon, preparation of medicine coated on medicine balloon and preparation method of medicine balloon

Technical Field

The invention belongs to the technical field of materials of catheters or coated catheters, and particularly relates to a drug balloon, preparation of a coated drug of the drug balloon and a preparation method of the drug balloon.

Background

Drug eluting balloon catheters are the product of a combination of traditional balloon angioplasty and advanced drug eluting techniques. The undegradability of the metal drug stent and the late-stage vascular thrombosis and inflammatory reaction caused by the metal drug stent prompt people to think about intervention non-implanted angioplasty, and the drug balloon is generated under the background. The drug eluting balloon catheter releases the drug immediately during interventional therapy, avoids side reaction caused by long-term retention of the metal stent and the polymer carrier, and is an effective supplement for metal stent angioplasty. The main technology of the currently marketed Sequent Please drug balloon is derived from the paccocath technology developed by Bruno Scheller and the like, a German medical doctor, paclitaxel is taken as an active drug, iopromide is taken as a matrix, and an organic solvent is added to the matrix for dissolving, and then the paclitaxel is coated on the surface of the balloon. The latter is a drug balloon technology with urea as a carrier and paclitaxel as an active drug by invatec; the euroco company adopts a coating technology with shellac as a carrier and paclitaxel as an active drug; the Lutonix company uses polysorbate as a carrier and paclitaxel as an active drug. Almost all drug balloons use paclitaxel as the active drug, with the exception of the carrier technology. This is because paclitaxel is rapidly absorbed by vascular smooth muscle cells and is retained in the blood vessels for a long period of time (30 days), thereby inhibiting the proliferation of the intima of the blood vessels.

Although paclitaxel drug balloons have met with much success clinically and commercially, their drug toxicity has been of constant concern to experts in the relevant field. Katsanos et al published a Meta analysis on a drug-coated device of paclitaxel on JAHA that might increase the patient's risk of 2-and 5-year death, based on which the U.S. FDA issued a statement that alerting clinicians that paclitaxel-coated balloons and stents for treatment of femoral artery disease might increase the patient's risk of death. The safety of rapamycin has been clinically proven, and the application of rapamycin in drug stents has been successful. Currently, rapamycin drug balloons are the hot spot of current research and the future development direction. However, rapamycin is absorbed slowly in vascular tissues and has a short retention time, and thus it is difficult to achieve an effect of inhibiting restenosis.

Rapamycin is a hydrophobic macrolide immunosuppressant, which acts by inhibiting cell cycle stages G0 and G1 by binding to the corresponding immunophilin RMBP, blocking G1 from entering S-phase. Is a powerful immunosuppressant with low toxicity and is widely applied to transplantation operations. Rapamycin is found to be the most commonly used drug in drug eluting stents because it is also effective in inhibiting the proliferation and migration of vascular smooth muscle cells following vascular injury, thereby reducing the incidence of vascular restenosis.

The toxicity of active drugs in the paclitaxel drug coating balloon catheter is a non-negligible factor, and meanwhile, the particle size of the paclitaxel drug coating is generally large, and the paclitaxel drug coating is difficult to escape during crossing, so that far-end vascular embolism is caused.

Disclosure of Invention

The invention provides a preparation method of a medicine balloon coating medicine, namely a preparation method of rapamycin nanoparticles, and a process method for coating the nanoparticles on the surface of a balloon catheter.

The specific process for preparing the nano particles comprises the following steps:

in the first step, an organic phase solution of drug and polymer is prepared. Rapamycin and polymer are dissolved in organic solvent, and the rapamycin and the polymer are dissolved thoroughly by means of magnetic stirring or ultrasonic homogenization and the like. The polymer can be chitosan, gelatin, sodium alginate, albumin, polylactic acid (PLA), polyglycolic acid, polylactic-co-glycolic acid (PLGA), polycaprolactone, polymethyl methacrylate, poloxamer, etc.; the organic solvent may be dichloromethane, chloroform, acetone, ethyl acetate, etc.; the mass ratio of the medicine to the polymer is 1: 1-10; preferably 1: 2.5.

in the second step, an aqueous solution containing a surfactant and a stabilizer is prepared. The surfactant may be polysorbate, methylcellulose, polyvinyl alcohol, vinyl cellulose, cross-linked acrylate polymers, and the like. The stabilizer can be carbomer, polyvinyl alcohol, etc. The proportion of the surfactant is 0.1-10%, and the proportion of the stabilizer is 0.1-5%. The volume ratio of the organic phase to the water phase is 1: 1-30, and preferably 1: 10.

thirdly, dropwise adding the organic phase solution into the aqueous phase solution, and simultaneously carrying out high-shear stirring, homogenization or ultrasonic treatment and the like on the aqueous phase solution to fully emulsify the aqueous phase solution; the organic phase can be added into the magnetically stirred aqueous phase solution at a uniform speed by using a precision injection pump or a dropping funnel, wherein the injection speed is 0.1 ml/min-1 ml/min, and preferably 0.3 ml/min; the magnetic stirring speed is 100-1000 rpm; preferably 400 rpm. An ultrasonic emulsification instrument can also be used for replacing magnetic stirring, and the ultrasonic power is as follows: 5-250W, working frequency: 19-25KHz, and 30-60 minutes of emulsification time.

Fourthly, removing the organic solvent to obtain a nano particle suspension of the polymer coated drug; continuously stirring the nanoparticle suspension for 6-30 hours by using a magnetic stirrer, and completely volatilizing the organic solvent to obtain a nanoparticle emulsion; or drying for 1-5 hours by using a vacuum drying oven, and removing an organic phase; alternatively, the organic phase was removed using a rotary evaporator.

Fifthly, centrifuging the nanoparticle suspension prepared in the fourth step by using a low-temperature high-speed centrifuge and cleaning by using high-purity water; and (4) subpackaging the nanoparticle suspension in a centrifuge tube, and putting the centrifuge tube into a centrifuge for centrifugation. Controlling the temperature to be 0-10 ℃, and preferably 4 ℃; the centrifugal speed is 3000-23000 r/m, preferably 10000-15000 r/m; removing supernatant from the centrifuged sample tube, adding deionized water for resuspension and centrifugation, and washing away free drugs and residual organic solvent; washing with water for 3-5 times.

Sixthly, freeze-drying the nano particles; the freeze-drying temperature is-50 to-80 ℃, and the vacuum degree is less than 10 Pa. The freeze-drying time is 10-48 hours. The step of freeze-drying the nano particles has breakthrough significance, guarantees the water-free property of the nano particles, and guarantees that the rapamycin is sprayed on the drug balloon. In the pharmaceutical preparation industry, the nanoparticles are generally prepared into injection preparations, and the injection preparations are injected into veins to be absorbed by tissues. The nano particles are prepared into solid nano particles required by intravascular devices, and the nano particles keep the original shape and structure after freeze-drying and wrap the medicinal components.

A preparation method of a rapamycin nanoparticle coated drug balloon is to prepare nanoparticles, then mix and disperse the nanoparticles and a vascular adhesive by using an organic solvent, and ultrasonically spray a dispersion liquid on the surface of the balloon, thereby obtaining a nanoparticle coated drug balloon catheter.

The blood vessel adhesive is phospholipid, cholesterol, fatty acid, etc. the phospholipid includes natural phospholipid selected from egg yolk lecithin, cephalin, soybean phospholipid, phosphatidylethanolamine, etc. and synthetic phospholipid selected from dipalmitoyl-DL- α -phosphatidylcholine (DPPC), Phosphatidylserine (PS), dimyristoyl lecithin (DMPC), Stearamide (SA), etc. the fatty acid is selected from lauroleic acid, tetradecadienoic acid, octanoic acid, myristic acid, decenoic acid, palmitoleic acid, linolenic acid, linoleic acid, stearic acid, etc.

Vascular adhesion agents refer to charged cationic liposomes, (2, 3-dioleoyl-propyl) -trimethylamine (DOTAP), N- [1- (2, 3-dioleoyl) propyl ] -N, N-trimethylammonium chloride (DOTMA), DC cholesterol, and the like. The cationic liposome coating can ensure that the nano particles are adhered to the surface of the vessel wall, thereby achieving the curative effect of slow release of the medicament. The cationic liposome blood vessel adhesive can adhere drug nanoparticles to the blood vessel wall. Because the blood vessel has a negative charge, the nanoparticles can adhere strongly to the vessel wall, and the nanoparticles have sufficient time to be absorbed by the vessel wall. The biological membrane of vascular smooth muscle cells is composed of phospholipid bilayers, hydrophilic phosphate groups are on the outside, and the phosphate groups have negative charges. Cationic liposomes, such as DOTAP, have a positive charge at the amino group of the molecule, and can bind to phosphate groups of cell membranes to form a strong binding force.

Dissolving the vascular adhesive in an organic solvent, and then adding the nanoparticles into the solution for suspension; the organic solvent does not dissolve the nanoparticles. The organic solvent is selected from pentane, hexane, a mixture of heptane and fluorocarbon, and the like.

Adding the nano particles into a solution in which the vascular adhesive is dissolved, dispersing by using ultrasonic, and spraying the nano particles onto the surface of the balloon.

The invention of the technical scheme is characterized in that rapamycin nanoparticles are prepared and used for coating medicine on a medicine balloon, and charged cationic liposome coating is adopted to block the medicine nanoparticles. The nano particles can be quickly absorbed by blood vessels, the slow release performance is provided, the retention time of the medicine in tissues is prolonged, the cationic liposome coating can ensure that the nano particles are adhered to the surface of the blood vessel wall, and the long-time medicine release is ensured.

Description of the drawings:

FIG. 1: the particle size of PLGA nanoparticles in example 1 was determined.

FIG. 2: scanning electron micrographs of the drug coating in example 1.

FIG. 3: the results of the drug effect test data of example 2.

Detailed Description

The technical solution is specifically illustrated according to the attached figures 1-3.

Example 1

Weighing 0.1g of rapamycin and 1g of PLGA, putting the rapamycin and the PLGA into a beaker containing 30 ml of acetone, and magnetically stirring the rapamycin and the PLGA for dissolving for later use; 3g of polyvinyl alcohol (PVA) was weighed out, added to 300ml of purified water, and dissolved by heating to 70 ℃ with a magnetic stirrer. And dropwise adding acetone into the magnetically stirred PVA aqueous solution at a stirring speed of 1500 rpm and a dropping speed of 0.5ml/min, and emulsifying for 2 minutes by using an ultrasonic probe emulsifier after the dropwise addition is finished.

And (3) centrifuging the emulsion at a high speed of 15000 r/min for 30 minutes, and washing with water for 3 times to obtain the PLGA nano particle suspension. The suspension was lyophilized for use. The freeze-drying temperature is-50 ℃, and the vacuum degree is less than 10 Pa. The time is 48 hours. A lyophilized nanoparticle powder was obtained.

Resuspending a small amount of nanoparticle powder in water, and performing particle size analysis with a laser scattering particle size analyzer (Mastersizer 3000, British Marvin) to obtain an average particle size of 138 nm, to obtain nanoparticle characterization, wherein the specific detection data result is shown in FIG. 1. The nanoparticles were observed using a scanning electron microscope, and the photograph thereof is shown in FIG. 2.

0.5g of phospholipid E80 (lipod, Germany) and 0.1g of cholesterol were dissolved in 20ml of n-heptane solvent while adding 1g of PLGA nanoparticles to the n-heptane solvent, and ultrasonically mixed for 2 minutes to obtain a spray-coated drug solution. Spraying the medicinal solution on a balloon catheter on an ultrasonic spraying machine. The injection flow is 0.6 ml/min, the moving speed of the spray head is 5.4mm/s, and the spraying times are 6 times.

And (4) airing the sprayed drug balloon, cutting, and shooting the drug crystallization form by using a scanning electron microscope. As can be seen in FIG. 2, the coating surface also has a large number of PLGA nanoparticles, which are bonded to each other by phospholipid and cholesterol.

Pharmacokinetic studies of drug balloons were performed using a domestic pig coronary model. Spraying medicine carrying amount of medicine saccule to 3ug/mm²And a protective sleeve is sleeved after folding and winding. Animals were tested after sterilization. The drug balloon is partially expanded on left coronary artery LAD, right coronary artery RCA and circumflex branch CX of the domestic pig, the balloon is expanded for 30-60 seconds, and then the balloon is withdrawn. At different time points, the animals are killed, the blood vessels are dissected, the dilated blood vessels are taken out, and the blood vessels are sent to a detection mechanism for detection. Animals were divided into 5 groups, and three vessels of 1 animal were treated per group, and sacrificed for dissection at 5 minutes, 7 days, 28 days, 60 days, and 90 days, respectively. The results of comparing the obtained tissue drug concentration data with those of similar products in the market are shown in fig. 3: at 5 minutes after administration, the tissue drug concentration of a control group (similar drug balloon on the market) is 262 mug/g, and the tissue drug concentration of the technical scheme is 385 mug/g; at 7 days after administration, the tissue drug concentration of a control group (similar drug balloon on the market) is 44 mug/g, and the tissue drug concentration of the technical scheme is 189 mug/g; at 28 days after administration, the tissue drug concentration of a control group (similar drug balloon on the market) is 21 mug/g, and the tissue drug concentration of the technical scheme is 80 mug/g; at 60 days after administration, control group: (Similar drug saccule in the market) tissue drug concentration is 19 mug/g, and the tissue drug concentration of the technical scheme is 9 mug/g; at 90 days after administration, the tissue drug concentration of a control group (similar drug balloon on the market) is 0 mug/g, and the tissue drug concentration of the technical scheme is 0 mug/g. It can be seen that the drug absorption rate of the drug balloon in the experimental group is obviously higher than that of the drug balloon in the control group in the prior art.

Example 2

In this example, the types and methods of the drugs and substances used are the same as in example 1, and the preferred ratio of the invention content is selected only in terms of the ratio of the drugs and substances, that is, the mass ratio of the drugs to the polymer in the first step is 1: 2.5; the volume ratio of the organic phase to the aqueous phase in the second step is 1: 10; the third step is that the injection speed is 0.3 ml/min; the magnetic stirring speed is 400 r/min; the fifth step is that the temperature is 4 ℃; the centrifugation speed was 12500 rpm. Other preparation processes are the same as example 1, and are not repeated herein, and the effect of this example is illustrated by pharmacokinetic studies.

Pharmacokinetic studies of drug balloons were performed using a domestic pig coronary model. Spraying medicine carrying amount of medicine saccule to 3ug/mm²And a protective sleeve is sleeved after folding and winding. Animals were tested after sterilization. The drug balloon is partially expanded on left coronary artery LAD, right coronary artery RCA and circumflex branch CX of the domestic pig, the balloon is expanded for 30-60 seconds, and then the balloon is withdrawn. At different time points, the animals are killed, the blood vessels are dissected, the dilated blood vessels are taken out, and the blood vessels are sent to a detection mechanism for detection. Animals were divided into 5 groups, and three vessels of 1 animal were treated per group, and sacrificed for dissection at 5 minutes, 7 days, 28 days, 60 days, and 90 days, respectively. The results of comparing the obtained tissue drug concentration data with those of similar products in the market are shown in fig. 3: at 5 minutes after administration, the tissue drug concentration of a control group (similar drug balloon on the market) is 262 mug/g, and the tissue drug concentration of the technical scheme is 462 mug/g; at 7 days after administration, the tissue drug concentration of a control group (similar drug balloon on the market) is 44 mug/g, and the tissue drug concentration of the technical scheme is 231 mug/g; at 28 days after administration, the tissue drug concentration of a control group (similar drug balloon on the market) is 21 mug/g, and the tissue drug of the technical schemeThe concentration is 83 mug/g; at 60 days after administration, the tissue drug concentration of a control group (similar drug balloon on the market) is 19 mug/g, and the tissue drug concentration of the technical scheme is 31 mug/g; at 90 days after administration, the tissue drug concentration of a control group (similar drug balloon on the market) is 0 mug/g, and the tissue drug concentration of the technical scheme is 0 mug/g. It can be seen that the drug absorption rate of the drug balloon in the experimental group is obviously higher than that of the drug balloon in the control group in the prior art.

Although the present invention has been described in detail with reference to the specific embodiments thereof, it should be understood by those skilled in the art that these are merely illustrative and not restrictive of the preferred embodiments, and that various other forms of the product may be made without departing from the spirit and scope of the invention, and that any changes in the form or structure may be made without departing from the spirit and scope of the invention.

Claims

1. A preparation method of a medicine balloon coated medicine is characterized by comprising the following steps:

(1) preparing an organic phase solution of the drug and the polymer;

(2) preparing an aqueous phase solution containing a surfactant and a stabilizer;

(3) adding the organic phase solution into the aqueous phase solution, stirring the aqueous phase solution with high shear, homogenizing or ultrasonically emulsifying the aqueous phase solution fully;

(4) removing the organic solvent to obtain a nanoparticle suspension of the polymer-coated drug, completely volatilizing the organic solvent to obtain a nanoparticle emulsion, and removing the organic phase;

(5) centrifuging the nanoparticle suspension prepared in the step (4), discarding supernatant in a centrifuged sample tube, adding deionized water for resuspension and centrifuging, and washing away free drugs and residual organic solvent;

(6) the nanoparticles were lyophilized.

2. The method of claim 1, wherein the balloon coating comprises a plurality of layers of the balloon coating, wherein the method comprises the steps of: the drug is rapamycin, and the mass ratio of the rapamycin to the polymer is 1:1 to 10.

3. The method for preparing a drug balloon-coated drug according to claim 1 or 2, characterized in that: the surfactant in the step (2) is polysorbate and/or methyl cellulose and/or polyvinyl alcohol and/or vinyl cellulose and/or a cross-linked acrylate polymer, and the proportion of the surfactant is 0.1% -10%.

4. The method of claim 3, wherein the balloon coating drug is prepared by: the stabilizer in the step (2) is carbomer and/or polyvinyl alcohol, and the proportion of the stabilizer is 0.1-5%.

5. The method for preparing a drug balloon-coated drug according to claim 4, wherein: in the step (3), the volume ratio of the organic phase to the water phase is 1: 1-30.

6. The preparation method of the medicine balloon is characterized by comprising the following steps:

(1) dissolving the vascular adhesive in an organic solvent;

(2) adding the nano particles into the solution for suspension;

(3) using ultrasonic dispersion, spray coating to the balloon surface.

7. The method of claim 6, wherein the method comprises the steps of: vascular adhesives are charged cationic liposomes.

8. The method of manufacturing a drug balloon according to claim 6 or 7, wherein: the vascular adhesive is (2, 3-dioleoyl-propyl) -trimethylamine (DOTAP) and/or N- [1- (2, 3-dioleoyl) propyl ] -N, N, N-trimethylammonium chloride (DOTMA) and/or DC cholesterol.

9. The method for preparing a drug balloon according to claim 8, wherein the method comprises the following steps: the organic solvent in step (1) is pentane and/or hexane and/or heptane and/or a fluorocarbon.

10. A drug balloon made using the method of claim 8.