Subdural hematoma targeted drug delivery system as well as preparation method and pharmaceutical preparation thereof

Abstract

The invention provides a subdural hematoma targeted drug delivery system, a preparation method thereof and a pharmaceutical preparation, and belongs to the technical field of targeted drugs. The targeted drug delivery system is a drug-loaded liposome, and the drug-loaded liposome comprises a drug active substance and a liposome carrier loaded with the drug active substance; the liposome carrier is prepared from the following components: cholesterol, lecithin, DSPE-PEG (Distearoyl Phosphate Polyethylene-Polyethylene Glycol) and DSPE-PEG-CLTX. The targeted drug delivery system provided by the invention is spherical, small and uniform in particle size, negatively charged on the surface and good in serum stability, can deliver the drug to the subdural hematoma part in a targeted manner, reduces systemic toxicity of the drug, inhibits pyroptosis, relieves inflammatory response, promotes hematoma absorption and meningeal lymphatic vessel drainage, and has a good application prospect. And the purpose of accurate treatment is achieved.

Description

Technical Field

[0001] This invention belongs to the field of targeted drug technology, specifically relating to a subdural hematoma targeted drug delivery system, its preparation method, and drug formulation. Background Technology

[0002] Subdural hematoma (SDH) is a common type of intracranial hematoma in clinical practice, with high rates of disability and mortality. It is mostly caused by ruptured bridging veins due to brain trauma. The hematoma accumulates in the subdural space, adversely affecting neurological function, manifesting as increased intracranial pressure and brain tissue displacement. If the hematoma persists for a long time, it may trigger inflammatory angiogenesis in the subdural space, leading to continuous bleeding and eventually developing into chronic subdural hematoma (CSDH). Therefore, timely removal of the subdural hematoma is crucial for the treatment of SDH.

[0003] Currently, the main clinical treatments for SDH include surgery, drug therapy (such as glucocorticoids and atorvastatin), supportive care, and rehabilitation. Drug therapy, however, has drawbacks such as insufficient drug targeting, a single mechanism of action, and significant side effects. For example, oral glucocorticoids have severe and long-term systemic side effects; atorvastatin has shown significant hepatotoxicity in some clinical patients.

[0004] Therefore, how to improve the therapeutic effect of drugs on SDH and reduce toxic side effects is an urgent problem to be solved in this field. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the present invention aims to provide a subdural hematoma targeted drug delivery system, its preparation method, and the drug formulation. This targeted drug delivery system can deliver drugs to the subdural hematoma site, reducing systemic drug toxicity, inhibiting pyroptosis, alleviating inflammatory responses, promoting hematoma absorption, and facilitating meningeal lymphatic drainage, thereby achieving the goal of precise treatment.

[0006] To achieve this objective, the present invention adopts the following technical solution:

[0007] In a first aspect, the present invention provides a targeted drug delivery system for subdural hematoma, wherein the targeted drug delivery system is a drug-loaded liposome, and the drug-loaded liposome includes a pharmaceutically active substance and a liposome carrier loaded with the pharmaceutically active substance;

[0008] The liposome carrier comprises cholesterol, lecithin, DSPE-PEG, and DSPE-PEG-CLTX.

[0009] DSPE: Distearate phosphatidylethanolamine; PEG: Polyethylene glycol; CLTX: Chlorotoxin; DSPE-PEG: PEG with DSPE attached to one end; DSPE-PEG-CLTX: PEG with DSPE and CLTX attached to both ends respectively. The pharmaceutically active substance referred to here is a pharmaceutically active substance used to treat subdural hematoma.

[0010] The targeted drug delivery system provided by this invention is a drug-loaded liposome, which is formed by the self-assembly of cholesterol, lecithin, DSPE-PEG, DSPE-PEG-CLTX, and active pharmaceutical ingredients. This drug-loaded liposome is spherical, with a small and uniform particle size, a negatively charged surface, and good serum stability.

[0011] The liposome carrier in the targeted drug delivery system provided by this invention is modified with CLTX. CLTX can selectively bind to matrix metalloproteinase-2 (MMP-2) upregulated in the hematoma region, thereby targeting the loaded drug active substance to the subdural hematoma site (including hematoma fluid and the meninges of the hematoma region). At the same time, the negative surface charge also makes it easier for the drug-loaded liposomes to target the weakly acidic hematoma tissue containing positive charge, thereby increasing the local drug concentration, reducing systemic toxicity, inhibiting pyroptosis and local inflammation caused by hematoma stimulation, promoting hematoma absorption and meningeal lymphatic drainage, and achieving the purpose of precise treatment.

[0012] In some embodiments of the present invention, the mass ratio of cholesterol, DSPE-PEG, DSPE-PEG-CLTX, and lecithin is (6-10):(50-70):(5-10):(8-12); for example, it can be 6:70:5:12, 6:70:10:8, 6:70:8:10, 6:60:8:10, 6:60:5:12, 6:60:10:8, 6:50:10:8, 6:50:5:12, 6:50:8:10, 8:70:5:12 ... The ratios are 10:8, 8:70:8:10, 8:60:8:10, 8:60:5:12, 8:60:10:8, 8:60:10:10, 8:50:10:8, 8:50:5:12, 8:50:8:10, 10:70:5:12, 10:70:10:8, 10:70:8:10, 10:60:8:10, 10:60:5:12, 10:60:10:8, 10:50:10:8, 10:50:5:12, or 10:50:8:10, etc. However, this invention is not limited to the listed values; other unlisted values ​​within this range are also applicable.

[0013] In this invention, controlling the proportions of each raw material within the aforementioned range helps to further reduce the particle size of drug-loaded liposomes, improve uniformity, and enhance stability. DSPE-PEG and DSPE-PEG-CLTX are the main structural components of the liposome carrier, with DSPE-PEG-CLTX providing targeting capability. Insufficient DSPE-PEG-CLTX results in poor targeting efficacy. Cholesterol helps stabilize the liposome structure and prevent structural breakage. However, cholesterol is a lipid-soluble substance, as are atorvastatin and dexamethasone. Excessive cholesterol content in these drugs can lead to larger liposome particle sizes, reducing the fusion efficiency between the liposomes and the cell membrane and hindering drug release. Lecithin helps reduce liposome particle size, but excessive lecithin can loosen the liposome membrane structure, increasing membrane permeability. This makes it easier for the drug encapsulated within the liposomes to leak out, resulting in significant drug loss before reaching the site of action during drug delivery, thus reducing therapeutic efficacy.

[0014] In some embodiments of the present invention, the weight-average molecular weight of PEG in the DSPE-PEG and the DSPE-PEG-CLTX is independently 1500-3000; for example, it can be 1500, 1800, 2000, 2200, 2500, 2800, or 3000, etc. However, the present invention is not limited to the listed values, and other unlisted values ​​within this range are also applicable.

[0015] In some embodiments of the present invention, the content of the active pharmaceutical ingredient in the drug-loaded liposome is 5-20 wt%; for example, it can be 5 wt%, 6 wt%, 8 wt%, 10 wt%, 12 wt%, 13 wt%, 15 wt%, 16 wt%, 18 wt%, or 20 wt%. However, the present invention is not limited to the listed values, and other unlisted values ​​within this range are also applicable.

[0016] In some embodiments of the present invention, the pharmaceutically active substances include atorvastatin (Ato) and dexamethasone (Dex). Atorvastatin is a first-line lipid-lowering drug in clinical practice, and dexamethasone is an oral glucocorticoid.

[0017] In some embodiments of the present invention, the mass ratio of atorvastatin to dexamethasone is 10:1.

[0018] In some embodiments of the present invention, the drug-loaded liposomes D 50The particle size is 100-200 nm; for example, it can be 100 nm, 110 nm, 120 nm, 130 nm, 140 nm, 150 nm, 160 nm, 170 nm, 180 nm, 190 nm, or 200 nm, etc. However, the present invention is not limited to the listed values, and other unlisted values ​​within this range are also applicable.

[0019] In a second aspect, the present invention provides a method for preparing a targeted drug delivery system as described in the first aspect, the method comprising the following steps:

[0020] Cholesterol, lecithin, DSPE-PEG, DSPE-PEG-CLTX and the active pharmaceutical ingredient are dissolved in an organic solvent to obtain an oil phase solution, which is then mixed with water. The raw materials self-assemble to form drug-loaded liposomes, which is the targeted drug delivery system.

[0021] In some embodiments of the present invention, the organic solvent is dimethyl sulfoxide (DMSO).

[0022] In some embodiments of the present invention, the concentration of the oil phase solution is 0.1-0.3 mg / mL; for example, it can be 0.1 mg / mL, 0.12 mg / mL, 0.15 mg / mL, 0.18 mg / mL, 0.2 mg / mL, 0.22 mg / mL, 0.25 mg / mL, 0.28 mg / mL, or 0.3 mg / mL, etc. However, the present invention is not limited to the listed values, and other unlisted values ​​within this range are also applicable.

[0023] In some embodiments of the present invention, the volume ratio of the oil phase solution to the water is 1:(3-8); for example, it can be 1:3, 1:3.3, 1:3.5, 1:3.8, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, 1:6.5, 1:7, 1:7.5, or 1:8, etc. However, the present invention is not limited to the listed values, and other unlisted values ​​within this range are also applicable.

[0024] In some embodiments of the present invention, the self-assembly conditions are as follows: the process is carried out under stirring at a stirring speed of 800-1500 rpm (e.g., 800 rpm, 900 rpm, 1000 rpm, 1100 rpm, 1200 rpm, 1300 rpm, 1400 rpm, or 1500 rpm, etc.), at a temperature of 20-37°C (e.g., 20°C, 22°C, 25°C, 28°C, 30°C, 32°C, 35°C, or 37°C, etc.), and for a time of 20-60 min (e.g., 20 min, 25 min, 30 min, 35 min, 40 min, 45 min, 50 min, 55 min, or 60 min, etc.). However, the present invention is not limited to the listed values; other unlisted values ​​within this range are also applicable.

[0025] In some embodiments of the present invention, the preparation method further includes: after the self-assembly is completed, dialysis of the product is performed to remove free raw materials.

[0026] In this invention, no special restrictions are placed on the preparation method of the DSPE-PEG-CLTX, and those skilled in the art can choose conventional methods. As a non-limiting example, the following method can be used for preparation:

[0027] DSPE-PEG-PDP (distearate phosphatidylethanolamine-polyethylene glycol-mercaptopyridine) was dissolved in DMSO, and CLTX was added. The reaction was carried out for 108 h under light-protected, magnetically stirred, and room temperature conditions. The reaction solution was then transferred to a dialysis bag and dialyzed in pure water under magnetic stirring for 24 h. The dialysate was collected and freeze-dried to obtain the product DSPE-PEG-CLTX.

[0028] Thirdly, the present invention provides a pharmaceutical preparation for treating subdural hematoma, the pharmaceutical preparation comprising a targeted drug delivery system as described in the first aspect, or a targeted drug delivery system prepared by the preparation method described in the second aspect.

[0029] In some embodiments of the present invention, the pharmaceutical preparation further comprises pharmaceutically acceptable excipients.

[0030] In some embodiments of the present invention, the dosage form of the pharmaceutical preparation is an injection.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] The targeted drug delivery system provided by this invention is a drug-loaded liposome, which is spherical, small and uniform in size, negatively charged on the surface, and has good serum stability. The drug-loaded liposome is modified with CLTX, which can selectively bind to matrix metalloproteinase-2 upregulated in the hematoma region, thereby targeting the loaded drug active substance to the subdural hematoma site (including hematoma fluid and the meninges in the hematoma region). Simultaneously, the negative surface charge makes it easier for the drug-loaded liposome to target the positively charged, weakly acidic hematoma tissue, thereby increasing the local drug concentration, reducing systemic toxicity, inhibiting pyroptosis and local inflammation caused by hematoma stimulation, promoting hematoma absorption and meningeal lymphatic drainage, and achieving the goal of precise treatment of SDH. Attached Figure Description

[0033] Figure 1 DSPE-PEG in this embodiment of the invention 2000 - CLTX synthesis roadmap;

[0034] Figure 2 This is a schematic diagram illustrating the synthesis of the drug-loaded liposomes prepared in Example 1 of the present invention;

[0035] Figure 3 The hydrodynamic diameter distribution diagrams are shown for the drug-loaded liposomes prepared in Example 1 and Comparative Example 1 of this invention.

[0036] Figure 4 The surface potential diagrams are of the drug-loaded liposomes prepared in Example 1 and Comparative Example 1 of this invention.

[0037] Figure 5 Transmission electron microscopy images of drug-loaded liposomes prepared in Example 1 and Comparative Example 1 of this invention;

[0038] Figure 6 The graphs show the particle size changes of the drug-loaded liposomes prepared in Examples 1-3 and Comparative Example 1 in the serum stability experiment.

[0039] Figure 7A These are in vivo fluorescence images of rats in each group during the targeting ability test of this invention.

[0040] Figure 7B This is a graph showing the quantitative fluorescence data of rats in each group during the targeting ability test of this invention;

[0041] Figure 7C This is a quantitative fluorescence data graph of hematoma fluid and meninges of rats in each group during the targeting ability test of this invention;

[0042] Figure 8A These are in vivo fluorescence images of rats in each group during the SDH treatment efficacy test of this invention.

[0043] Figure 8B This is a graph showing the quantitative fluorescence data of rats in each group during the SDH treatment efficacy test of this invention. Detailed Implementation

[0044] The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments. Those skilled in the art should understand that the specific embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention.

[0045] DSPE-PEG used in the embodiments of the present invention 2000 -CLTX synthesis route is as follows Figure 1 As shown, the specific preparation method is as follows:

[0046] Take 10mg of DSPE-PEG 2000 - PDP was dissolved in 3 mL of DMSO, and 5 mg of CLTX was added. The mixture was reacted for 108 h under light-protected conditions, with magnetic stirring at 800 rpm and at room temperature. The reaction solution was then transferred to a dialysis bag and dialyzed against pure water at 300 rpm with magnetic stirring for 24 h. The dialysate was collected, freeze-dried, and the product DSPE-PEG was obtained.2000 -CLTX;

[0047] Among them, PEG 2000 This refers to PEG with a weight-average molecular weight of 2000.

[0048] Example 1

[0049] This embodiment provides a subdural hematoma targeted drug delivery system, which is a drug-loaded liposome, comprising a drug-active substance and a liposome carrier loaded with the drug-active substance;

[0050] The active pharmaceutical ingredients are atorvastatin and dexamethasone; the liposome carriers consist of cholesterol, lecithin, and DSPE-PEG. 2000 and DSPE-PEG 2000 -CLTX.

[0051] The schematic diagram of the synthesis of drug-loaded liposomes described in this embodiment is shown below. Figure 2 As shown, the specific preparation method is as follows:

[0052] Take 3mg DSPE-PEG 2000 -CLTX, 3mg DSPE-PEG 2000 1 mg cholesterol, 1 mg lecithin, 10 mg atorvastatin and 1 mg dexamethasone were dissolved in 1 mL of dimethyl sulfoxide to obtain solutions of the corresponding 5 substances;

[0053] According to cholesterol, DSPE-PEG 2000 DSPE-PEG 2000 The mass ratio of CLTX, lecithin, atorvastatin, and dexamethasone was 8:60:10:10:10:1. Taking the above solution, 4 μL, 10 μL, 1.67 μL, 5 μL, 0.5 μL, and 0.5 μL of the above raw material solution, respectively, and adding them to dimethyl sulfoxide (total volume controlled at 300 μL), the mixture was stirred at 1500 rpm for 5 min at room temperature to obtain an oil phase solution. This oil phase solution was then added to 1 mL of ddH2O stirred magnetically at 1500 rpm. After stirring for 60 min, the mixture was transferred to a dialysis bag with a molecular weight cutoff of 3500 D and dialyzed at 200 rpm for 12 h to obtain drug-loaded liposomes (denoted as C / D / L-Ato@Dex, where C refers to DSPE-PEG). 2000 -CLTX, D refers to DSPE-PEG 2000 L refers to lecithin, Ato refers to atorvastatin, and Dex refers to dexamethasone.

[0054] Example 2

[0055] This embodiment provides a subdural hematoma targeted drug delivery system, which is a drug-loaded liposome. The only difference from Example 1 is that it does not contain dexamethasone and instead uses the same mass of cholesterol. This drug-loaded liposome is designated C / D / L-Ato.

[0056] Example 3

[0057] This embodiment provides a subdural hematoma targeted drug delivery system, which is a drug-loaded liposome. The only difference from Example 1 is that it does not contain atorvastatin and instead contains the same mass of cholesterol. This drug-loaded liposome is denoted as C / D / L-Dex.

[0058] Example 4

[0059] This embodiment provides a targeted drug delivery system for subdural hematoma, which is a drug-loaded liposome, and the preparation method is as follows:

[0060] Take 3mg DSPE-PEG 2000 -CLTX, 3mg DSPE-PEG 2000 1 mg cholesterol, 1 mg lecithin, 10 mg atorvastatin and 1 mg dexamethasone were dissolved in 1 mL of dimethyl sulfoxide to obtain solutions of the corresponding 5 substances;

[0061] According to cholesterol, DSPE-PEG 2000 DSPE-PEG 2000 The mass ratio of CLTX, lecithin, atorvastatin, and dexamethasone was 6:70:5:12:10:1. The above solution was prepared by taking 3 μL, 11.67 μL, 0.83 μL, 6 μL, 0.5 μL, and 0.5 μL of the above raw material solution, respectively, and adding them to dimethyl sulfoxide (total volume controlled at 300 μL). The mixture was stirred at 1500 rpm for 5 min at room temperature using a magnetic stirrer to obtain an oil phase solution. This oil phase solution was then added to 1.5 mL of ddH2O, which was magnetically stirred at 800 rpm. After stirring for 40 min, the mixture was transferred to a dialysis bag with a molecular weight cutoff of 3500 D and dialyzed at 200 rpm for 12 h to obtain drug-loaded liposomes.

[0062] Example 5

[0063] This embodiment provides a targeted drug delivery system for subdural hematoma, which is a drug-loaded liposome, and the preparation method is as follows:

[0064] Take 3mg DSPE-PEG 2000 -CLTX, 3mg DSPE-PEG 20001 mg cholesterol, 1 mg lecithin, 10 mg atorvastatin and 1 mg dexamethasone were dissolved in 1 mL of dimethyl sulfoxide to obtain solutions of the corresponding 5 substances;

[0065] According to cholesterol, DSPE-PEG 2000 DSPE-PEG 2000 The mass ratio of CLTX, lecithin, atorvastatin, and dexamethasone was 10:50:8:8:10:1. The above solution was prepared by taking 5 μL, 8.33 μL, 1.33 μL, 4 μL, 0.5 μL, and 0.5 μL of the above raw material solution, respectively, and adding them to dimethyl sulfoxide (total volume controlled at 300 μL). The mixture was stirred at 1500 rpm for 5 min at room temperature to obtain an oil phase solution. This oil phase solution was then added to 2.4 mL of ddH2O stirred magnetically at 1000 rpm. After stirring for 20 min, the mixture was transferred to a dialysis bag with a molecular weight cutoff of 3500D and dialyzed at 200 rpm for 12 h to obtain drug-loaded liposomes.

[0066] Comparative Example 1

[0067] This comparative example provides a non-targeted drug-loaded liposome, which is prepared by the following method:

[0068] Take 3mg DSPE-PEG 2000 1 mg cholesterol, 1 mg lecithin, 10 mg atorvastatin and 1 mg dexamethasone were dissolved in 1 mL of dimethyl sulfoxide to obtain solutions of the corresponding 5 substances;

[0069] According to cholesterol, DSPE-PEG 2000 The mass ratio of lecithin, atorvastatin, and dexamethasone was 8:70:10:10:1. Specifically, 4 μL, 11.67 μL, 5 μL, 0.5 μL, and 0.5 μL of the above raw material solutions were respectively added to dimethyl sulfoxide (total volume controlled at 300 μL). The mixture was stirred at 1500 rpm for 5 min at room temperature using a magnetic stirrer to obtain an oil phase solution. This oil phase solution was then added to 1 mL of ddH2O under magnetic stirring at 1500 rpm. After stirring for another 60 min, the mixture was transferred to a dialysis bag with a molecular weight cutoff of 3500 D and dialyzed at 200 rpm for 12 h to obtain drug-loaded liposomes (denoted as D / L-Ato@Dex).

[0070] Comparative Example 2

[0071] This comparative example provides a drug-loaded liposome, which differs from Example 1 only in that it contains cholesterol and DSPE-PEG. 2000 DSPE-PEG 2000- The mass ratio of CLTX to lecithin is 15:60:10:10, which means the amount of cholesterol solution used is 7.5 μL.

[0072] Comparative Example 3

[0073] This comparative example provides a drug-loaded liposome, which differs from Example 1 only in that it contains cholesterol and DSPE-PEG. 2000 DSPE-PEG 2000 The mass ratio of CLTX to lecithin is 8:60:10:15, meaning the amount of lecithin solution used is 7.5 μL.

[0074] Performance testing

[0075] 1. Hydrodynamic diameter and surface potential testing

[0076] The hydrodynamic diameter, PDI (dispersion index), and surface potential of the drug-loaded liposomes prepared in the above examples and comparative examples were measured using a Malvern particle size analyzer. The test results are shown in Table 1. The hydrodynamic diameter distributions of C / D / L-Ato@Dex and D / L-Ato@Dex prepared in Example 1 and Comparative Example 1 are shown in Table 1. Figure 3 As shown, where A is C / D / L-Ato@Dex and B is D / L-Ato@Dex; the surface potential is as follows. Figure 4 As shown.

[0077] Table 1

[0078]

[0079]

[0080] From Table 1 and Figure 3 , Figure 4 As can be seen, the drug-loaded liposomes prepared in the embodiments of the present invention have an average hydrodynamic diameter of less than 200 nm, a PDI of less than 0.2, and small and uniform particle size; and their surface is negatively charged, which is beneficial for dispersion in blood circulation and makes them less likely to aggregate; and they are more likely to be targeted to weakly acidic hematoma tissue containing positive charge.

[0081] Compared with Example 1, the amount of cholesterol added in Comparative Example 2 was higher, and the amount of lecithin added in Comparative Example 3 was higher. Both of these resulted in larger particle sizes of drug-loaded liposomes, which would reduce the fusion efficiency of liposomes with cell membranes and hinder drug release.

[0082] 2. Morphological characteristics

[0083] The morphology of C / D / L-Ato@Dex and D / L-Ato@Dex prepared in Example 1 and Comparative Example 1 was observed using transmission electron microscopy. The results are as follows: Figure 5 As shown.

[0084] Figure 5 The results show that the C / D / L-Ato@Dex prepared in Example 1 is a uniformly dispersed spherical shape with a diameter of less than 200 nm.

[0085] 3. Serum stability test

[0086] The drug-loaded liposomes prepared in Examples 1-3 and Comparative Example 1 were incubated for 7 days in PBS buffer (pH = 7.4) containing 10 vol% FBS (fetal bovine serum). The particle size changes of the drug-loaded liposomes were continuously detected using a Malvern particle size analyzer. The results are as follows: Figure 6 As shown.

[0087] Figure 6 The results showed that the particle size of the drug-loaded liposomes prepared in Examples 1-3 and Comparative Example 1 did not differ significantly within 7 days, indicating that the nanoparticles did not undergo significant deformation, thus demonstrating their good serum stability.

[0088] 4. Targeting capability test

[0089] The solutions of C / D / L-Ato@Dex prepared in Example 1 and D / L-Ato@Dex prepared in Comparative Example 1 were respectively mixed with DMSO solution of DiR (a lipophilic fluorescent dye) to prepare solutions of appropriate concentrations. The solutions were stirred thoroughly and transferred to 3500D dialysis bags and dialyzed overnight in water in the dark to obtain solutions of DiR-labeled drug-loaded liposomes, which were denoted as DiR-C / D / L-Ato@Dex and DiR-D / L-Ato@Dex, respectively.

[0090] A rat model of SDH (subdural hemorrhage) was established by subdural injection of autologous blood. Rats were anesthetized with isoflurane (5% induction, 2% maintenance). Hair on the top of the head was shaved and the scalp was carefully disinfected. A 1 cm incision was made along the midline to expose the dorsal side of the skull. A bone window, 1.5 mm in diameter and 1 mm deep, was drilled 3 mm to the right of the posterior fontanelle. The dura mater was carefully torn using microforceps, and 400 μL of autologous blood from the femoral vein was injected into the subdural space at a rate of 50 μL / min using a microinfusion pump equipped with a 14G indwelling needle. The indwelling needle was left in place for 10 minutes and then removed to prevent leakage. The bone window was sealed with medical adhesive, and the scalp incision was disinfected and sutured. Postoperatively, ibuprofen (10 mg / kg) was administered intraperitoneally to relieve pain.

[0091] After successful establishment of the SDH rat model, rats were randomly divided into three groups of three. Atorvastatin (3 mg / kg) and dexamethasone (0.3 mg / kg) were administered via tail vein injection of DiR-C / D / L-Ato@Dex, DiR-D / L-Ato@Dex, and DiR, respectively. Rats in each group were sacrificed at 6, 12, 24, and 48 hours post-administration. The head skin and soft tissue were dissected, and fluorescence distribution and intensity were detected using in vivo fluorescence imaging. Hematoma fluid and meninges from the hematoma area were collected, homogenized, and their fluorescence content was detected using an ELISA reader.

[0092] Figure 7A These are in vivo fluorescence images of rats in each group. Figure 7B The graph shows the quantitative fluorescence data of rats in each group. Figure 7C The image shows the quantitative fluorescence data of hematoma fluid and meninges of rats in each group.

[0093] Experimental results showed that the fluorescence intensity of the hematoma area in the C / D / L-Ato@Dex group rats was significantly higher than that in the DiR-D / L-Ato@Dex group and the DiR group rats, indicating that the drug delivery system of the present invention has good targeting ability of SDH.

[0094] 5. SDH treatment efficacy test

[0095] Rats were randomly divided into 8 groups: Sham group, SDH group, Ato group, Dex group, Ato+Dex group, C / D / L-Ato group, C / D / L-Dex group, and C / D / L-Ato@Dex group.

[0096] Except for the Sham group, rats were used to establish an SDH rat model according to the aforementioned method. In this model, autologous blood from the femoral vein was centrifuged to separate erythrocytes, which were then labeled with DiR and injected into the subdural space of the rats. Rats in the Sham group underwent sham surgery: only the skull was opened, and no autologous blood was injected.

[0097] After the surgery, the rats in each group were treated for 3 days as follows:

[0098] Sham group: Normal saline was administered by gavage;

[0099] SDH group: Normal saline was administered by gavage;

[0100] Ato group: Atorvastatin was administered by gavage at a dose of 3 mg / kg / day;

[0101] Dex group: Dexamethasone was administered by gavage at a dose of 0.3 mg / kg / day;

[0102] Ato+Dex group: Atorvastatin 3 mg / kg·d and dexamethasone 0.3 mg / kg·d were administered by gavage;

[0103] C / D / L-Ato group: C / D / L-Ato prepared in Example 2 was administered via tail vein injection at a dosage of atorvastatin 3 mg / kg·d;

[0104] C / D / L-Dex group: C / D / L-Dex prepared in Example 3 was administered via tail vein injection at a dosage of dexamethasone 0.3 mg / kg·d;

[0105] C / D / L-Ato@Dex group: C / D / L-Ato@Dex prepared in Example 1 was administered via tail vein injection at the dosage of atorvastatin 3 mg / kg·d and dexamethasone 0.3 mg / kg·d.

[0106] Three days after treatment, the fluorescence distribution and intensity of each group were detected by in vivo fluorescence imaging in small animals to determine the hematoma absorption status of the SDH rat model.

[0107] Figure 8A These are in vivo fluorescence images of rats in each group. Figure 8B The graph shows the quantitative fluorescence data of rats in each group.

[0108] Experimental results showed that, compared with the SDH group, the fluorescence intensity of the hematoma area in each drug-treated group decreased to varying degrees. The fluorescence intensity in the C / D / L-Ato group was significantly lower than that in the Ato group, the C / D / L-Dex group was significantly lower than that in the Dex group, and the C / D / L-Ato@Dex group was significantly lower than that in the Ato+Dex group. Furthermore, the fluorescence intensity in the hematoma area was lowest in the C / D / L-Ato@Dex group. This indicates that the drug delivery system of the present invention can effectively promote hematoma absorption and improve the therapeutic effect of the drug on SDH by targeting the drug to the hematoma area. Moreover, the therapeutic effect of the combination of atorvastatin and dexamethasone is superior to that of the single-drug therapy.

[0109] The above description is merely a specific embodiment of this disclosure, enabling those skilled in the art to understand or implement it. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of this disclosure. Therefore, this disclosure is not to be limited to the embodiments described herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A subdural hematoma targeted drug delivery system, characterized in that, The targeted drug delivery system is a drug-loaded liposome, which includes a drug-active substance and a liposome carrier loaded with the drug-active substance. The liposome carrier comprises cholesterol, lecithin, DSPE-PEG, and DSPE-PEG-CLTX.

2. The targeted drug delivery system according to claim 1, characterized in that, The mass ratio of cholesterol, DSPE-PEG, DSPE-PEG-CLTX and lecithin is (6-10):(50-70):(5-10):(8-12).

3. The targeted drug delivery system according to claim 1 or 2, characterized in that, The weight-average molecular weight of PEG in both DSPE-PEG and DSPE-PEG-CLTX is independently 1500-3000.

4. The targeted drug delivery system according to any one of claims 1-3, characterized in that, The content of the active pharmaceutical ingredient in the drug-loaded liposome is 5-20 wt%. Preferably, the active pharmaceutical ingredient includes atorvastatin and dexamethasone; Preferably, the mass ratio of atorvastatin to dexamethasone is 10:

1.

5. The targeted drug delivery system according to any one of claims 1-4, characterized in that, The drug-loaded liposomes D 50 The particle size is 100-200nm.

6. A method for preparing a targeted drug delivery system as described in any one of claims 1-5, characterized in that, The preparation method includes the following steps: Cholesterol, lecithin, DSPE-PEG, DSPE-PEG-CLTX and the active pharmaceutical ingredient are dissolved in an organic solvent to obtain an oil phase solution, which is then mixed with water. The raw materials self-assemble to form drug-loaded liposomes, which is the targeted drug delivery system.

7. The preparation method according to claim 6, characterized in that, The organic solvent is dimethyl sulfoxide; Preferably, the concentration of the oil phase solution is 0.1-0.3 mg / mL; Preferably, the volume ratio of the oil phase solution to the water is 1:(3-8); Preferably, the self-assembly conditions are as follows: the self-assembly is carried out under stirring at a stirring speed of 800-1500 rpm, a temperature of 20-37℃, and a time of 20-60 min.

8. The preparation method according to claim 6 or 7, characterized in that, The preparation method further includes: after the self-assembly is completed, dialysis is performed on the product to remove free raw materials.

9. A pharmaceutical preparation for treating subdural hematoma, characterized in that, The pharmaceutical preparation comprises the targeted drug delivery system as described in any one of claims 1-5, or the targeted drug delivery system prepared by the preparation method as described in any one of claims 6-8.

10. The pharmaceutical preparation according to claim 9, characterized in that, The pharmaceutical preparation also contains pharmaceutically acceptable excipients; Preferably, the dosage form of the pharmaceutical preparation is an injection.

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