Crystal form A of rolimus medicine, preparation method of crystal form A, medicine composition and medicine coating balloon

Abstract

The invention relates to the field of medicines, and provides a crystal form A of a limus medicine, a preparation method of the crystal form A, a medicine composition and a medicine coating balloon. The crystal form A of the limus medicine is radiated by Cu-Kalpha, is diffracted by X-rays expressed by a 2 theta angle, and has characteristic peaks at 10.2 + / -0.2 degrees, 13.3 + / -0.2 degrees, 14.3 + / -0.2 degrees and 21.7 + / -0.2 degrees. The crystal form A is uniform in structure, good in stability and beneficial to long-time storage; the sustained-release tablet has the advantages of good lipophilicity, low dissolution rate, long residence time in target blood vessels, and realization of continuous sustained release.

Description

Technical Field

[0001] The present invention relates to the field of medicines, and in particular to a crystal form A of a limus drug, a preparation method thereof, a pharmaceutical composition and a drug-coated balloon. Background Art

[0002] The morbidity and mortality of diseases such as atherosclerosis caused by vascular stenosis are rising year by year. Arterial stenting is the main means of treating coronary heart disease at present. However, with the increase in the number of stents used, it is very easy to cause postoperative vascular restenosis, which seriously affects the treatment effect. The restenosis rate of lesions after simple balloon dilatation is as high as 27%. There is an urgent need for the emergence of devices that can deliver drugs to target vessels without foreign body implantation. Drug-coated balloons (DCBs) have been proven to be an effective measure to deal with vascular restenosis and primary lesions of small coronary vessels and large coronary vessels. Its emergence provides a new option for the treatment of coronary artery disease. As a new interventional treatment technology, it has been gradually widely used in the field of coronary artery and peripheral intervention worldwide. DCB is a traditional balloon covered with a layer of anti-proliferative drugs. Its drug components can quickly penetrate into the vascular wall within a single balloon dilatation to inhibit the proliferation and migration of smooth muscle cells, thereby hindering the process of restenosis. Compared with drug-eluting stents (DES), DCB has no metal mesh residue, which reduces the inflammatory response of the intima, greatly reduces the risk of thrombosis, and can shorten the duration of dual antiplatelet therapy (only 1-3 months of dual antiplatelet therapy is required after DCB surgery). At the same time, DCB treatment avoids the placement of foreign bodies, leaving patients with the opportunity for follow-up treatment when necessary.

[0003] To date, there are more than ten DCB products on the market worldwide, most of which use paclitaxel-based drug coatings. Paclitaxel has good lipid solubility and stable anti-proliferative effects. It effectively inhibits the proliferation of smooth muscle cells by stabilizing microtubule assembly and blocking mitosis. However, due to the mechanism of paclitaxel cytotoxicity, long-term embolism caused by the shedding of coating particles, and vascular calcification, paclitaxel-coated balloons still have potential risks.

[0004] Compared with paclitaxel, limus drugs and their derivatives are more mature in drug-eluting stents (DES), and the safety and efficacy of the drugs used are supported by reliable clinical evidence. Compared with paclitaxel-coated balloons, limus drugs and their derivatives (such as sirolimus, tacrolimus, euclidinium, everolimus, etc.) coated balloons have stronger anti-proliferative effects in preventing restenosis, including anti-inflammatory protection, as well as a wider range of treatment and a larger safety margin. Studies have shown that compared with paclitaxel-eluting stents, limus drugs and their derivatives eluting stents show lower inflammatory response and less biotin deposition. However, due to the high hydrophilicity of limus drugs, their drug retention time in the vascular wall is short, and the difficulty in sustained release is the main challenge for the clinical transformation of limus drugs.

[0005] In summary, improving the lipophilicity of limus drugs, increasing the retention time of limus drugs in target blood vessels, and prolonging the sustained release time of limus drugs are of great significance for further enhancing the ability of limus drugs to inhibit vascular stenosis. Summary of the invention

[0006] Therefore, the technical problem to be solved by the present invention is to overcome the defects of the prior art of limus drugs, such as poor lipophilicity, short retention time in target blood vessels, and difficulty in sustained release, thereby providing a crystalline form A of limus drugs, a preparation method thereof, a pharmaceutical composition, and a drug-coated balloon.

[0007] To this end, the present invention provides the following technical solutions:

[0008] The invention provides a crystal form A of a limus drug, which uses Cu-Kɑ radiation and X-ray diffraction represented by 2θ angles, and has characteristic peaks at 10.2±0.2°, 13.3±0.2°, 14.3±0.2° and 21.7±0.2°.

[0009] In an optional embodiment, the crystal form A of the limus drug uses Cu-Kα radiation and the X-ray diffraction expressed as 2θ angles also has characteristic peaks at one or more of 12.7±0.2°, 16.2±0.2°, 16.9±0.2°, 20.1±0.2°, and 20.7±0.2°.

[0010] In an optional embodiment, the crystal form A of the limus drug uses Cu-Kɑ radiation and the X-ray diffraction expressed as 2θ angles also has characteristic peaks at one or more of 15.5±0.2°, 18.2±0.2°, 18.6±0.2°, 19.7±0.2°, 23.9±0.2°, 24.5±0.2°, and 25.7±0.2°.

[0011] In an optional embodiment, the crystal form A of the limus drug uses Cu-Kα radiation, and the X-ray diffraction expressed as 2θ angle has two characteristic peaks between 14.2° and 14.4°.

[0012] In an optional embodiment, the crystal form A of the limus drug uses Cu-Kα radiation and X-ray diffraction expressed as 2θ angles, and also has a characteristic peak at 28.0±0.2°.

[0013] In an optional embodiment, the crystal form A of the limus drug uses Cu-Kα radiation and X-ray diffraction expressed as 2θ angles, and also has a characteristic peak at 25.2±0.2°.

[0014] In an optional embodiment, the limus drugs include one or more of sirolimus, tacrolimus, everolimus, ugripilimus, gustationilimus, pimecrolimus and temsirolimus.

[0015] In an optional embodiment, the crystal form A is a rod-shaped or needle-shaped structure.

[0016] The rod-like or needle-like structure helps the crystal form A to be inserted into the target blood vessel wall, so that the drug can exert its effect in the target blood vessel for a long time.

[0017] Optionally, the crystal form A has a length of 1-50 μm and a width of 100-1200 nm.

[0018] Optionally, the crystal conversion rate of the crystal form A is ≥70%.

[0019] The present invention also provides a method for preparing the crystalline form A of the limus drug, comprising the following steps:

[0020] The limus drug and the good solvent are first mixed to obtain a solution;

[0021] The solution is mixed with a poor solvent for a second time until crystals are precipitated to obtain a crystalline form A of the limus drug.

[0022] A good solvent refers to a solvent that has good solubility for limus drugs.

[0023] In an optional embodiment, the good solvent includes one or more of acetonitrile, dimethyl sulfoxide, ethanol, methanol, acetone, chloroform, ethyl acetate, diethyl ether and dimethylformamide.

[0024] Optionally, the mass ratio of the limus drug to the good solvent is (0.03-0.50):1.

[0025] Optionally, the temperature of the first mixing is 10-35° C., the rotation speed is 400-1200 rpm, and after the first mixing is sufficient, a solution is obtained.

[0026] Poor solvents refer to solvents that are slightly soluble or poorly soluble in limus drugs.

[0027] In an optional embodiment, the poor solvent includes one or more of water, n-heptane, petroleum ether, dichloromethane, tetrahydrofuran, ethylene glycol, hexane, butyl acetate and isopropanol.

[0028] Optionally, the volume ratio of the poor solvent to the good solvent is (1-50):1.

[0029] A poor solvent is added to the solution under stirring to perform a second mixing, and the second mixing is performed until crystals are precipitated.

[0030] In an optional embodiment, the stirring temperature is 10-35° C., and the stirring speed is 400-1200 rpm.

[0031] Optionally, the temperature of the second mixing is 10-35° C., and the rotation speed is 400-1200 rpm.

[0032] Optionally, the second mixing includes a third mixing, and the third mixing temperature is 10-35° C., the rotation speed is 400-1200 rpm, and the time is 8-24 hours.

[0033] Through the preparation method of the present invention, the limus drugs are converted from an amorphous state to a crystalline state, and the crystal conversion rate is ≥70%.

[0034] The present invention also provides a pharmaceutical composition, comprising the crystal form A of the limus drug or the crystal form A of the limus drug prepared by the preparation method, and pharmaceutically acceptable excipients.

[0035] The present invention also provides a drug-coated balloon, comprising a balloon body and a drug coating coated on the balloon body, wherein the drug coating comprises the crystalline form A of the limus drug or the crystalline form A of the limus drug obtained by the preparation method.

[0036] The technical solution of the present invention has the following advantages:

[0037] 1. The present invention provides a crystal form A of a limus drug, which uses Cu-Kɑ radiation and X-ray diffraction expressed in 2θ angles, and has characteristic peaks at 10.2±0.2°, 13.3±0.2°, 14.3±0.2°, and 21.7±0.2°. The crystal form A has a uniform structure, good stability, and is conducive to long-term storage.

[0038] 2. The slender rod-like or needle-like structure is conducive to the insertion of drugs into the vascular villi and endothelial cells, increasing the retention time of drugs in the target blood vessels, achieving long-term sustained release and repair of drugs in the target blood vessels, and reducing the restenosis rate of blood vessels.

[0039] 3. The present invention also provides a method for preparing Form A of a sirolimus drug, which comprises the following steps: performing a first mixing of the sirolimus drug and a good solvent to obtain a solution; performing a second mixing of the solution with a poor solvent until crystals precipitate to obtain Form A of the sirolimus drug. The preparation method is simple in operation, does not require the separate introduction of a nucleating agent during the process, has low requirements for process equipment, and the product size is uniform and controllable. The obtained Form A has good lipophilicity, a low dissolution rate, and a relatively long residence time in the target blood vessel, and can achieve sustained release. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0041] Figure 1 It is a morphological characterization diagram of crystallized everolimus in Example 1 of the present invention at a scale of 300 μm;

[0042] Figure 2 It is a morphological characterization diagram of crystallized everolimus in Example 1 of the present invention at a scale of 8 μm;

[0043] Figure 3 It is an XRD diffraction pattern (Intensity - intensity) of everolimus raw material in Example 1 of the present invention;

[0044] Figure 4 It is an XRD diffraction pattern (Intensity - intensity) of crystallized everolimus in Example 1 of the present invention;

[0045] Figure 5 It is a morphological characterization diagram of crystallized everolimus in Example 2 of the present invention at a scale of 300 μm;

[0046] Figure 6 It is a morphological characterization diagram of crystallized everolimus in Example 2 of the present invention at a scale of 8 μm;

[0047] Figure 7 It is an XRD diffraction pattern (Intensity - intensity) of crystallized everolimus in Example 2 of the present invention;

[0048] Figure 8 It is a morphological characterization diagram of crystallized everolimus in Example 3 of the present invention at a scale of 300 μm;

[0049] Fig. 9This is a morphology characterization diagram of crystallized everolimus in Example 3 of the present invention at a scale of 8 μm;

[0050] Fig.10 is the XRD diffraction pattern (Intensity) of crystallized everolimus in Example 3 of the present invention;

[0051] Fig.11 is a comparison chart of the cumulative release rate of everolimus API and crystallized everolimus over time in Example 1 of the present invention;

[0052] Fig.12 Schematic diagram of in vitro test results of different drug-loaded balloons in Experimental Example 2 of the present invention;

[0053] Fig.13 This is a morphological characterization diagram of the crystallized everolimus in Example 1 of the present invention at an instantaneous time and a scale of 15 μm;

[0054] Fig.14 This is a morphological characterization diagram of the crystallized everolimus in Example 1 of the present invention at day 3 with a scale of 15 μm;

[0055] Fig.15 This is a morphological characterization diagram of the crystallized everolimus in Example 1 of the present invention at day 7 with a scale of 15 μm. DETAILED DESCRIPTION

[0056] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0057] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0058] Example 1

[0059] The limus drug (everolimus API) is first mixed with acetonitrile and ethanol, wherein the mass ratio of the limus drug, acetonitrile and ethanol is 0.04:0.33:0.67, the temperature of the first mixing is 25° C., the rotation speed is 700 rpm, and after the first mixing is sufficient, a solution is obtained;

[0060] Under stirring, the stirring temperature is 25°C, the rotation speed is 700 rpm, water is added to the solution, the volume ratio of water to the aforementioned acetonitrile is 7:1, and the second mixing is continued, the temperature of the second mixing is 25°C, the rotation speed is 700 rpm, the second mixing is carried out until crystals precipitate, and the third mixing is carried out, the temperature of the third mixing is 25°C, the rotation speed is 700 rpm, and the time is 15 hours to obtain Form A of everolimus (referred to as crystallized everolimus).

[0061] The crystallized everolimus prepared in this example was subjected to morphological characterization, and a morphological characterization diagram of the crystallized everolimus in this example at a scale of 300 μm was obtained, as shown in FIG. Figure 1 As shown; the morphology characterization diagram of the crystallized everolimus in this embodiment at a scale of 8 μm, as shown Figure 2 As shown. Figure 1 and 2 It can be seen that the crystallized everolimus prepared in this example is a rod-shaped structure with a uniform crystal structure, a length of 2-20 μm, and a width of 400-900 nm.

[0062] The crystal conversion rate of crystallized everolimus was tested as follows: the initial everolimus drug (everolimus raw material) was weighed and recorded as m 1 , m 1 =45 mg; the system obtained after the third mixing is filtered, and then the solid obtained by filtration is dried and weighed, which is recorded as m 2 , m 2 =35mg. Crystal conversion rate = m 2 (35mg) / m 1 (45 mg)*100%=77.78%.

[0063] The everolimus API and the crystallized everolimus prepared in this example were irradiated with Cu-Ka radiation to obtain XRD diffraction patterns of the everolimus API. Figure 3 As shown; the XRD diffraction pattern of crystallized everolimus in this embodiment is as shown Figure 4 The crystal form data of the crystallized everolimus in this embodiment are shown in Table 1. Figure 3-4 As can be seen from Table 1, the crystallized limus drug has multiple obvious and easily distinguishable characteristic diffraction peaks, while the non-crystallized drug crystals have only one very broad diffraction peak, and its structural characteristics cannot be distinguished.

[0064] Table 1 Crystal form data of crystallized everolimus in Example 1

[0065] Peak number 2θ(°) Relative strength (%) 1 10.208 36.41 2 12.660 42.31 3 13.344 14.2 4 14.281 100 5 14.348 62.02 6 15.541 11.41 7 16.229 30.72 8 16.883 39.59 9 18.179 21.74 10 18.630 12.74 11 19.722 16.68 12 20.149 30.08 13 20.656 30.99 14 21.725 27.13 15 23.930 13.93 16 24.463 16.16 17 25.737 15.36

[0066] Example 2

[0067] The limus drug (everolimus raw material) and ethanol are first mixed, the mass ratio of the limus drug to the ethanol is 0.19:1, the temperature of the first mixing is 25° C., the rotation speed is 700 rpm, and after the first mixing is sufficient, a solution is obtained;

[0068] Under stirring, the stirring temperature is 25°C, the rotation speed is 700 rpm, petroleum ether is added to the solution, the volume ratio of petroleum ether to the aforementioned ethanol is 15:1, and the second mixing is continued, the temperature of the second mixing is 25°C, the rotation speed is 700 rpm, the second mixing is carried out until crystals precipitate, and the third mixing is carried out, the temperature of the third mixing is 25°C, the rotation speed is 700 rpm, and the time is 15 hours to obtain Form A of everolimus (referred to as crystallized everolimus).

[0069] The crystallized everolimus prepared in this example was subjected to morphological characterization, and a morphological characterization diagram of the crystallized everolimus in this example at a scale of 300 μm was obtained, as shown in FIG. Figure 5 As shown; the morphology characterization diagram of the crystallized everolimus in this embodiment at a scale of 8 μm, as shown Figure 6 As shown. Figure 5 and 6 It can be seen that the crystallized everolimus prepared in this example is a rod-shaped structure with a uniform crystal structure, a length of 2-5 μm, and a width of 200-590 nm.

[0070] According to the method of Example 1, the crystal conversion rate = m 2 (41mg) / m 1 (50 mg)*100%=82.00%.

[0071] The crystallized everolimus prepared in this example was irradiated with Cu-Ka radiation to obtain an XRD diffraction pattern of the crystallized everolimus in this example. Figure 7 The crystal form data of the crystallized everolimus in this embodiment are shown in Table 2. Figure 7 As can be seen from Table 2, the crystallized limus drug has multiple characteristic diffraction peaks that are obvious and easy to distinguish.

[0072] Table 2 Crystal form data of crystallized everolimus in Example 2

[0073] Peak number 2θ(°) Relative strength (%) 1 10.208 36.37 2 12.661 59.61 3 13.340 32.35 4 14.279 100 5 14.345 23.56 6 15.543 11.45 7 16.224 40.35 8 16.863 39.63 9 18.168 27.64 10 18.628 12.78 11 19.699 26.74 12 20.112 35.60 13 20.671 54.16 14 21.732 59.34 15 23.934 22.14 16 24.490 21.95 17 25.741 15.33 18 27.998 10.79

[0074] Example 3

[0075] The limus drug (everolimus API) and dimethyl sulfoxide are first mixed, the mass ratio of the limus drug to the dimethyl sulfoxide is 0.23:1, the temperature of the first mixing is 25° C., the rotation speed is 700 rpm, and after the first mixing is sufficient, a solution is obtained;

[0076] Under stirring, the stirring temperature is 25°C, the rotation speed is 700 rpm, water is added to the solution, the volume ratio of water to the aforementioned dimethyl sulfoxide is 6:1, and the second mixing is continued, the temperature of the second mixing is 25°C, the rotation speed is 700 rpm, the second mixing is carried out until crystals precipitate, and the third mixing is carried out, the temperature of the third mixing is 25°C, the rotation speed is 700 rpm, and the time is 13 hours to obtain Form A of everolimus (referred to as crystallized everolimus).

[0077] The crystallized everolimus prepared in this example was subjected to morphological characterization, and a morphological characterization diagram of the crystallized everolimus in this example at a scale of 300 μm was obtained, as shown in FIG. Figure 8 As shown; the morphology characterization diagram of the crystallized everolimus in this embodiment at a scale of 8 μm, as shown Fig. 9 As shown. Figure 8 and 9 It can be seen that the crystallized everolimus prepared in this example is a needle-shaped structure with a uniform crystal structure, a length of 3-12 μm, and a width of 300-600 nm.

[0078] According to the method of Example 1, the crystal conversion rate = m 2 (39mg) / m 1 (49 mg)*100%=79.59%.

[0079] The crystallized everolimus prepared in this example was irradiated with Cu-Ka radiation to obtain an XRD diffraction pattern of the crystallized everolimus in this example. Fig.10 The crystal form data of the crystallized everolimus in this embodiment are shown in Table 3. Fig.10 As can be seen from Table 3, the crystallized limus drug has multiple characteristic diffraction peaks that are obvious and easy to distinguish.

[0080] Table 3 Crystal form data of crystallized everolimus in Example 3

[0081]

[0082]

[0083] Experimental Example 1

[0084] 5 mg of the everolimus bulk drug used in Example 1 and 5 mg of the crystallized everolimus prepared in Example 1 were weighed, and dissolved in 50 mL of a 0.2% SDS (sodium dodecyl sulfate) solution, respectively. The particles were evenly dispersed by short-term ultrasound and then taken out immediately. They were then placed on a 37°C shaker at a speed of 70 rpm for sustained release dissolution. After the sampling time, 2 mL of the supernatant was taken to test its drug concentration, and 2 mL of a 0.2% SDS solution was added to continue the sustained release. According to the test method, the drug dissolution rate of the everolimus bulk drug and the crystallized everolimus was tested, and a comparison chart of the cumulative release rate of the everolimus bulk drug and the crystallized everolimus in Example 1 over time was obtained, as shown in FIG. Fig.11 shown. Fig.11 In this document, the API refers to the API of everolimus, and the drug crystal refers to the crystallized everolimus. Fig.11 It can be seen that the dissolution rate of crystallized everolimus is slower than that of amorphous everolimus API. In the early stage, the dissolution rate of everolimus API is 1.2-1.4 times that of crystallized everolimus, and in the later stage, the dissolution rates of the two are close. This indicates that after the drug is crystallized, the dissolution rate is reduced, which is conducive to the long-term sustained release of the drug in the tissue.

[0085] Experimental Example 2

[0086] Weigh 150 mg of the everolimus raw material used in Example 1 and 150 mg of the crystallized everolimus prepared in Example 1, disperse them in 10 mL of n-heptane respectively, stir for 3 hours at 20-25°C to obtain two mixed solutions. Then select a common balloon (PTCA balloon dilatation catheter S025010-4 of Shanghai Yingtai Medical Instrument Co., Ltd.), fill the balloon to 5 atm, maintain the temperature of the mixed solution at 20-25°C, control the liquid inlet flow rate of the inlet to 0.5 mL / min, the stirring speed of the stirring device on the inlet to 500 rpm, and an ultrasonic sprayer is provided at the outlet of the inlet, and the ultrasonic power is 1 W. By adjusting the number of spraying times 10 times, the mass density of the drug loaded on the balloon is controlled to be 2.0 μg / mm 2 , and two drug-loaded balloons were obtained.

[0087] Two prepared drug-loaded balloons were subjected to in vitro experiments: a silicone tube of appropriate size (drug-loaded balloon diameter: silicone tube inner diameter = 1.1-1.3) was selected to replace the target blood vessel, PBS solution (pH 7.4) was selected as simulated blood, the solution temperature was controlled to 37±2°C, the solution flow rate through the silicone tube was adjusted to 70mL / min, and the two drug-loaded balloons prepared from the crystallized everolimus and everolimus bulk drug in Example 1 were expanded to the balloon nominal pressure on the silicone tube, and the pressure was maintained for 60s. After the drug coating was fully transferred to the silicone tube, the balloon was withdrawn from the solution, and the silicone tube after the transfer operation was kept continuously flushed under the above conditions. The flushing time was set to 0 time, 1 day, 3 days, 5 days, etc. After a fixed time, the silicone tube of the drug coating transfer section was cut off, an appropriate amount of acetonitrile was added, and the drug concentration was tested by high-performance liquid chromatography. The residual amount of the drug on the silicone tube at different time periods was examined to characterize the in vitro sustained release time and effect of the drug coating. A schematic diagram of the in vitro test results of different drug-loaded balloons was obtained, as shown Fig.12 As shown. Fig.12 It can be seen that the drug coating on the silicone tube after the everolimus API drug-loaded balloon was transferred was no longer detectable at 1 day, and the drug coating on the silicone tube after the crystallized everolimus drug-loaded balloon was transferred was still 10.03% of the initial transfer value at 3 days, and no drug was detected at 5 days, indicating that the sustained-release time of crystallized everolimus is longer than that of the API.

[0088] Experimental Example 3

[0089] The crystallized everolimus prepared in Example 1 was stored in a refrigerator at 4°C, and the morphological characteristics of the crystallized drug were photographed using a scanning electron microscope after preparation, 3 days (24*3 hours), and 7 days (24*7 hours). Figure 13-15 As shown, they are the morphological characterization diagrams of the crystallized everolimus in Example 1 at different times (immediately, 3 days, 7 days) and a scale of 15 μm. Figure 13-15 It can be seen that the crystallized drug can maintain a normal morphology during the tested time, all of which are rod-shaped or needle-shaped structures with a length of 2-20 μm and a width of 400-900 nm, indicating that crystallized everolimus has good stability and is conducive to long-term storage.

[0090] Experimental Example 4

[0091] According to Experimental Example 2, the everolimus API used in Example 1 and the crystallized everolimus prepared in Example 1 were respectively prepared into drug-loaded balloons, and the balloon selected was the PTCA balloon dilatation catheter S030020-4 of Shanghai Yingtai Medical Instrument Co., Ltd., to obtain two types of drug-loaded balloons. The experiment selected 6 white pigs (labeled Z1, Z2, Z3, Z4, Z5, and Z6) as animal models, and 3 coronary arteries were selected for each white pig. The experiment was conducted at the vascular position matching the diameter of the drug-loaded balloon (the coronary artery diameter was measured by DSA imaging technology, and the diameter of the drug-loaded balloon was 1.1-1.3 times the diameter of the blood vessel), simulating the actual clinical PCI (percutaneous coronary intervention) surgery, and the blood vessels of Z1, Z2, and Z3 were given drug-loaded balloons corresponding to the everolimus API. , the drug-loaded balloon corresponding to crystallized everolimus was administered to the blood vessels of Z4, Z5, and Z6, and the drug-loaded balloon was expanded to the nominal pressure at the target blood vessel, and the pressure was maintained for 60 seconds before being withdrawn from the body. Three time points were set, namely, immediate (Z1 and Z4), 7 days (i.e., 7*24h, Z2 and Z5), and 14 days (i.e., 14*24h, Z3 and Z6). The corresponding target blood vessels were dissected during the corresponding follow-up period to test the tissue drug concentration, and the in vivo test results of different drug-loaded balloons were obtained, as shown in Table 4. From Table 4, it can be seen that the initial transfer of the drug-loaded balloon corresponding to crystallized everolimus in the vascular tissue was high, with an average drug concentration of 48.52μg / g. After 14 days, the remaining drug concentration in the vascular tissue was still 0.62μg / g, accounting for 1.28% of the initial average concentration. The initial transfer rate of the drug-loaded spheres corresponding to the raw material drug everolimus was relatively low, only half of the transfer rate of the crystallized everolimus drug-loaded balloons, and at 7 days, its tissue drug concentration dropped rapidly to only 1.97% of the initial average value, and the drug could not be detected after 14 days. This shows that the crystallized everolimus has a better transfer effect on vascular tissue and excellent long-term sustained release effect. This is closely related to the rod-like structure of the crystallized drug. The rod-like structure can effectively insert the drug into the tissue during balloon expansion, achieving high adhesion and long-term sustained release on the tissue.

[0092] Among them, the test method for vascular tissue drug concentration (μg / g, the mass of drug contained in each gram of vascular tissue) is as follows: weigh a certain amount of sample, add 1mL of acetonitrile and then add zirconium oxide grinding beads to grind for 10 minutes, then centrifuge at 13000rpm for 10 minutes at 4°C, and finally filter with a 0.22μm filter membrane, and take the filtrate for high performance liquid chromatography analysis.

[0093] Table 4 In vivo test results of different drug-loaded balloons

[0094]

[0095]

[0096] Note: BLQ means not detected.

[0097] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A crystalline form A of a limus drug, characterized in that: Using Cu-Kɑ radiation, X-ray diffraction expressed in 2θ angles showed characteristic peaks at 10.2±0.2°, 13.3±0.2°, 14.3±0.2°, and 21.7±0.2°.

2. The crystalline form A of the limus drug according to claim 1, characterized in that: Using Cu-Kɑ radiation, X-ray diffraction expressed in 2θ angles also has characteristic peaks at one or more of 12.7±0.2°, 16.2±0.2°, 16.9±0.2°, 20.1±0.2°, and 20.7±0.2°.

3. The crystalline form A of the limus drug according to claim 1, characterized in that: Using Cu-Kɑ radiation, X-ray diffraction expressed in 2θ angles also has characteristic peaks at one or more of 15.5±0.2°, 18.2±0.2°, 18.6±0.2°, 19.7±0.2°, 23.9±0.2°, 24.5±0.2°, and 25.7±0.2°.

4. The crystalline form A of the limus drug according to claim 1, characterized in that: The limus drugs include one or more of sirolimus, tacrolimus, everolimus, ugriplimus, gumarilimus, pimecrolimus and temsirolimus.

5. The crystalline form A of the limus drug according to claim 1, characterized in that: The crystal form A is a rod-shaped or needle-shaped structure; Optionally, the crystal form A has a length of 1-50 μm and a width of 100-1200 nm; Optionally, the crystal conversion rate of the crystal form A is ≥70%.

6. The method for preparing the crystalline form A of the limus drug according to any one of claims 1 to 5, characterized in that: It includes the following steps: The limus drug and the good solvent are first mixed to obtain a solution; The solution is mixed with a poor solvent for a second time until crystals are precipitated to obtain a crystalline form A of the limus drug.

7. The method for preparing the crystal form A of the limus drug according to claim 6, characterized in that: The good solvent includes one or more of acetonitrile, dimethyl sulfoxide, ethanol, methanol, acetone, chloroform, ethyl acetate, ether and dimethylformamide; Optionally, the mass ratio of the limus drug to the good solvent is (0.03-0.50):1; Optionally, the temperature of the first mixing is 10-35° C., and the rotation speed is 400-1200 rpm.

8. The method for preparing the crystal form A of the limus drug according to claim 6, characterized in that: The poor solvent includes one or more of water, n-heptane, petroleum ether, dichloromethane, tetrahydrofuran, ethylene glycol, hexane, butyl acetate and isopropanol; Optionally, the volume ratio of the poor solvent to the good solvent is (1-50):1; Optionally, the temperature of the second mixing is 10-35° C. and the rotation speed is 400-1200 rpm; Optionally, the second mixing further includes a third mixing, wherein the temperature of the third mixing is 10-35° C., the rotation speed is 400-1200 rpm, and the time is 8-24 hours.

9. A pharmaceutical composition, characterized in that The invention comprises the crystal form A of the limus drug described in any one of claims 1 to 5 or the crystal form A of the limus drug prepared by the preparation method described in any one of claims 6 to 8, and pharmaceutically acceptable excipients.

10. A drug-coated balloon, characterized in that: It comprises a balloon body and a drug coating coated on the balloon body, wherein the drug coating comprises the crystalline form A of the limus drug described in any one of claims 1-5 or the crystalline form A of the limus drug prepared by the preparation method described in any one of claims 6-8.

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