CN105056243B - A kind of pharmaceutical composition of the mesoporous copper sulfide of hyaluronic acid decorated magnetic hollow and preparation method and application - Google Patents

Abstract

The present invention relates to a hyaluronic acid-modified magnetic hollow mesoporous copper sulfide pharmaceutical composition and its preparation method and application. It can effectively solve the problem of good biocompatibility, strong magnetic targeting, and low toxic and side effects to improve the tumor treatment effect. The problem is that the method is to load anti-tumor drugs on the aminated copper sulfide obtained by the reaction of hollow mesoporous copper sulfide nanoparticles and mercaptoethylamine. The copper sulfide surface is electrostatically adsorbed on the ferric tetroxide nanoparticles, and reacts on the surface through amide bonds. Modify hyaluronic acid to form a pharmaceutical composition of hyaluronic acid-modified magnetic hollow mesoporous copper sulfide; the mass content ratio of ferric oxide and copper sulfide is 1 to 8:1, and the magnetic hollow mesoporous copper sulfide The particle size of the invention is 50~300nm. The method of the invention is easy to operate, the method is stable and reliable, and has the advantages of good biocompatibility, strong magnetic targeting, and low toxic and side effects. In terms of tumor treatment, it can play a role in hyperthermia, tumor diagnosis, and magnetic field buffering. Controlled release drugs and other functions.

Description

A pharmaceutical composition of magnetic hollow mesoporous copper sulfide modified by hyaluronic acid and its Preparation method and application

technical field

The invention relates to medicine, in particular to a pharmaceutical composition of magnetic hollow mesoporous copper sulfide modified by hyaluronic acid, a preparation method and application thereof.

Background technique

Surgery, radiotherapy, and chemotherapy are the three most commonly used means of tumor treatment. However, surgery can only remove tumors that are visible to the naked eye, but it is difficult to remove invisible subclinical lesions; traditional chemotherapy often has the problems of poor targeting and dose-limiting toxicity; new treatment methods. Photothermal therapy technology is a minimally invasive technology that uses photothermal conversion materials to convert the energy of near-infrared light into heat energy. Photothermal therapy can achieve local heating and kill cancer cells in the treatment of cancer, but it has no effect on normal cells. Damage is a safer and more efficient means of tumor treatment.

As a semiconductor crystal material, copper sulfide nanoparticles have strong near-infrared absorption and high light-to-heat conversion efficiency. Compared with other noble metal nanomaterials, its absorption wavelength in the infrared region is less affected by the environment and size. In addition, the latest report found that copper sulfide nanoparticles can generate active oxygen (including singlet oxygen, superoxide anions, etc.) under near-infrared light. These reactive oxygen species are highly reactive and can exhibit significant cytotoxicity. Copper sulfide is less toxic to organisms and can be degraded by organisms. It is a relatively safe material, so it has received extensive attention in the photothermal therapy of tumors.

The pore structure inside the functionalized magnetic hollow mesoporous copper sulfide enables it to have an ultra-high drug loading capacity. Its particle size is small, its physical and chemical properties are stable, and it has excellent magnetic targeting. By modifying its surface with water-soluble groups such as hyaluronic acid, it can significantly improve the biocompatibility and biocompatibility of magnetic hollow mesoporous copper sulfide in vivo. Fluidity is conducive to the direct delivery of drugs to the lesion under the condition of an external magnetic field. Since the ferroferric oxide also has the function of blocking holes in the carrier, it rotates and vibrates under the action of an alternating magnetic field and then deforms to open the holes, thereby achieving the effect of controlled drug release under the control of the magnetic field.

The chemotherapeutic drugs are loaded into the nano-carriers with photothermal effect, and are delivered to the tumor site through the EPR effect, so as to further improve the therapeutic effect and reduce the toxic and side effects on normal tissues and cells. The synthesis method of the carrier and its pharmaceutical composition is simple, and combines the remarkable photothermal effect of copper sulfide, the magnetic targeting of ferric oxide and its controlled release of drugs under the action of a magnetic field, and the good biocompatibility of hyaluronic acid. In addition, it can physically load chemotherapy drugs with anti-tumor activity, which has the advantages of high efficiency and controllability compared with traditional methods, and the combination of photothermal therapy and chemotherapy technology reflects the concept of comprehensive cancer treatment . Therefore, it is of great significance and value to study a targeted therapeutic agent that combines photothermal therapy and chemotherapy on the same nano-platform.

Contents of the invention

In view of the above situation, in order to overcome the defects of the prior art, the object of the present invention is to provide a hyaluronic acid-modified magnetic hollow mesoporous copper sulfide pharmaceutical composition and its preparation method and application, which can effectively solve the problem of poor biocompatibility. , Strong magnetic targeting, less toxic and side effects, so as to improve the effect of tumor treatment.

The technical scheme solved by the present invention is that the aminated copper sulfide obtained by the reaction of hollow mesoporous copper sulfide nanoparticles and mercaptoethylamine is loaded with antineoplastic drugs, and ferric oxide nanoparticles are electrostatically adsorbed on the surface of copper sulfide. The hyaluronic acid is reacted on the surface modification to form a pharmaceutical composition of magnetic hollow mesoporous copper sulfide modified by hyaluronic acid; The particle size of porous copper sulfide is 50~300nm, which is realized by the following steps:

(1) Dissolve 0.7-0.8g of CuSO ₄ ·5H ₂ O in 180-220ml of water, stir, add 5-7g of polyvinylpyrrolidone, react for 10-20min, add 1.1-1.3g of sodium hydroxide, stir evenly, Add 11-13ml of hydrazine hydrate dropwise, stir for 5-10min, add 1.2-1.4ml of ammonium sulfate, stir for 1h, centrifuge with ultrapure water at 12000-15000rpm (ie 12000-15000r/min) for 5-10min, wash with water until neutral, freeze Dry for 48 hours to obtain hollow mesoporous copper sulfide nanoparticles;

(2) Ultrasonically disperse 90-110mg hollow mesoporous copper sulfide nanoparticles in 90-110ml ultra-pure water, ultrasonicate for 10-30min, add 180-220mg mercaptoethylamine under stirring, stir for 24h, centrifuge at 12000-15000rpm for 5- 10min to get the precipitate, add ultra-pure water to redissolve the precipitate, then centrifuge at 12000-15000rpm for 5-10min to get the precipitate, add ultra-pure water to re-dissolve the precipitate, repeat this 2~3 times until it is neutral, freeze-dry, and obtain amination Hollow mesoporous copper sulfide composite (CuS-NH ₂ );

(3) Synthesis of ferroferric oxide nanoparticles: Take 0.8-0.9 g FeCl ₂ and 2.3-2.4 g FeCl ₃ and dissolve them in 35-45ml of water under stirring, heat to 80°C under nitrogen protection, slowly add 4-6ml of ammonia water dropwise , stirred in an oil bath for 30 minutes, added 1.8-2.2ml of citric acid with a mass concentration of 0.5g·ml ^-1 , raised the temperature to 95°C, reacted for 90 minutes, cooled to room temperature while stirring, dialyzed to remove unreacted substances, and freeze-dried to obtain water-soluble Fe3O4 nanoparticles;

(4) Take 4.5-5.5 mg of the aminated hollow mesoporous copper sulfide complex (CuS-NH ₂ ) obtained in step (2), disperse it in 4.5-5.5 ml of phosphate buffer solution with a pH of 7-9, and place it in an ice bath Ultrasonic for 10-30 min, mixed with phosphate buffer containing anti-tumor drugs, each 3ml of phosphate buffer contains 9g of anti-tumor drugs, stirred at room temperature for 24 hours, centrifuged and washed with water for 3 times, the probe was ultrasonically dispersed in water, freeze-dried Aminated copper sulfide loaded with antitumor drugs;

(5) Disperse the aminated copper sulfide loaded with antineoplastic drugs in 18-22ml of water, add iron ferric oxide nanoparticles under stirring, stir at room temperature for 6-48h, centrifuge at 12000-15000rpm for 5-10min to obtain a precipitate, freeze-dry, The magnetic hollow mesoporous copper sulfide loaded with antitumor drugs is obtained;

(6) Ultrasonically disperse the drug-loaded magnetic hollow mesoporous copper sulfide in 18-22ml of water, add 180-220mg of hyaluronic acid in aqueous solution, and stir for 6-48h to form a hyaluronic acid-modified magnetic hollow medium Pharmaceutical compositions of porous copper sulfide.

The invention is easy to operate, the method is stable and reliable, and the prepared copper sulfide and its pharmaceutical composition have the advantages of good biocompatibility, strong magnetic targeting, and small toxic and side effects, and can play a role in hyperthermia and tumor diagnosis in tumor treatment. , magnetic field slow and controlled drug release, etc., is an innovation in tumor treatment drugs, and has huge economic and social benefits.

detailed description

The specific implementation of the present invention will be described in detail below in conjunction with specific conditions and examples.

Example 1

In the specific implementation of the present invention, the preparation method of the pharmaceutical composition of magnetic hollow mesoporous copper sulfide modified by hyaluronic acid is realized by the following method:

(1) Dissolve 0.75g of CuSO ₄ ·5H ₂ O in 200ml of water, stir, add 6.0g of polyvinylpyrrolidone, react for 10~20min, add 1.2g of sodium hydroxide, stir well, add 12ml of hydration dropwise with a syringe Hydrazine, stirred for 5-10min, finally added 1.29ml of ammonium sulfate, stirred for 1h, centrifuged with ultrapure water at 15000rpm for 5-10min, washed until neutral, freeze-dried for 48h, and hollow mesoporous copper sulfide nanoparticles were obtained;

(2) Ultrasonically disperse hollow mesoporous copper sulfide nanoparticles 100mg probe in 100ml ultrapure water, ultrasonic for 10~30min, add 200mg of mercaptoethylamine under stirring condition, stir for 24h, centrifuge at 15000rpm for 5-10min to get precipitate, add super Redissolve in pure water, then centrifuge, and repeat this 2~3 times until it is neutral, freeze-dried to obtain an aminated hollow mesoporous copper sulfide composite (CuS-NH ₂ );

(3) Synthesis of ferroferric oxide nanoparticles: Take 0.86 g FeCl ₂ and 2.35 g FeCl ₃ and dissolve them in 40ml of water under stirring, heat to 80°C under the protection of nitrogen, slowly add 5ml of ammonia water dropwise with a syringe, stir in an oil bath for 30min, Then add 2ml of citric acid with a mass concentration of 0.5g ml ^-1 , heat up to 95°C, react for 90min, cool to room temperature under stirring, dialyze to remove unreacted substances, and freeze-dry to obtain water-soluble iron ferric oxide nanoparticles;

(4) Take 5 mg of aminated hollow mesoporous copper sulfide complex, disperse it in 5 ml of phosphate buffer, ultrasonicate the probe under ice bath for 10-30 min, and mix it with phosphate buffer containing antineoplastic drugs. The saline buffer contains 9g of anti-tumor drugs, stirred at room temperature for 24 hours, centrifuged at 15000rpm for 5-10 minutes, washed with water for 3 times, ultrasonically dispersed in water with the probe, and freeze-dried to obtain the amidated copper sulfide loaded with anti-tumor drugs;

(5) Disperse the amidated copper sulfide loaded with antineoplastic drugs in 20ml of water, add ferric oxide dispersed in water under agitation, stir at room temperature for 24 hours, centrifuge to obtain a precipitate, and freeze-dry to obtain antineoplastic drug loaded Magnetic hollow mesoporous copper sulfide;

(6) Ultrasonically disperse the drug-loaded magnetic hollow mesoporous copper sulfide probe in 20 ml of water, add 200 mg of hyaluronic acid aqueous solution, and stir for 24 hours to obtain a pharmaceutical composition of hyaluronic acid-modified magnetic hollow mesoporous copper sulfide.

Example 2

In the specific implementation of the present invention, the preparation method of the pharmaceutical composition of magnetic hollow mesoporous copper sulfide modified by hyaluronic acid is realized by the following method:

(1) Dissolve 0.7g of CuSO ₄ ·5H ₂ O in 180ml of water, stir, add 5g of polyvinylpyrrolidone, react for 10~20min, add 1.1g of sodium hydroxide, stir well, then add dropwise 11ml of hydrazine hydrate, stir 5~10min, add 1.2ml of ammonium sulfate, stir for 1h, centrifuge with ultrapure water at 12000rpm for 10min, wash until neutral, and freeze-dry for 48h to obtain hollow mesoporous copper sulfide nanoparticles;

(2) Ultrasonically disperse 90mg of hollow mesoporous copper sulfide nanoparticles in 90ml of ultrapure water, ultrasonically for 10~30min, add 180mg of mercaptoethylamine under stirring, stir for 24h, centrifuge at 12000rpm for 10min to obtain a precipitate, add ultrapure water to the precipitate Then centrifuge at 12000rpm for 10min to obtain a precipitate, add ultrapure water to redissolve the precipitate, and repeat this 2~3 times until it is neutral, freeze-dry to obtain an aminated hollow mesoporous copper sulfide complex (CuS-NH ₂ );

(3) Synthesis of Fe3O4 nanoparticles: Dissolve 0.8 g FeCl ₂ and 2.3 g FeCl ₃ in 35 ml of water under stirring, heat to 80°C under nitrogen protection, slowly add 4-6 ml of ammonia water dropwise, and stir in an oil bath for 30 min. Add 1.8ml of citric acid with a mass concentration of 0.5gml- ¹ , heat up to 95°C, react for 90min, cool to room temperature under stirring, dialyze to remove unreacted substances, and freeze-dry to obtain water-soluble iron ferric oxide nanoparticles;

(4) Take 4.5 mg of the aminated hollow mesoporous copper sulfide complex (CuS-NH ₂ ) obtained in step (2), disperse it in 4.5 ml of phosphate buffer solution with pH 7-9, and sonicate for 10~ 30 min, mixed with phosphate buffer containing anti-tumor drugs, each 3ml of phosphate buffer contains 9g of anti-tumor drugs, stirred at room temperature for 24 hours, centrifuged and washed 3 times with water, ultrasonically dispersed the probe in water, freeze-dried to obtain the loaded antibody Aminated copper sulfide for tumor drugs;

(5) Disperse the aminated copper sulfide loaded with antineoplastic drugs in 18ml of water, add ferric oxide nanoparticles under stirring, stir at room temperature for 6-48h, centrifuge at 12000rpm for 10min to obtain a precipitate, and freeze-dry to obtain the loaded antineoplastic drug magnetic hollow mesoporous copper sulfide;

(6) Ultrasonic disperse the above drug-loaded magnetic hollow mesoporous copper sulfide in 18ml of water, add 180mg of hyaluronic acid in aqueous solution, stir and react for 6-48h, and the magnetic hollow mesoporous copper sulfide modified by hyaluronic acid is obtained. pharmaceutical composition.

Example 3

In the specific implementation of the present invention, the preparation method of the pharmaceutical composition of magnetic hollow mesoporous copper sulfide modified by hyaluronic acid is realized by the following method:

(1) Dissolve 0.8g of CuSO ₄ ·5H ₂ O in 220ml of water, stir, add 7g of polyvinylpyrrolidone, react for 10~20min, add 1.3g of sodium hydroxide, stir well, then add 13ml of hydrazine hydrate dropwise, stir 5~10min, add 1.4ml of ammonium sulfate, stir for 1h, centrifuge with ultrapure water for 8min at 13000rpm, wash until neutral, and freeze-dry for 48h to obtain hollow mesoporous copper sulfide nanoparticles;

(2) Ultrasonically disperse 110 mg of hollow mesoporous copper sulfide nanoparticles in 110 ml of ultrapure water, ultrasonicate for 10-30 minutes, add 220 mg of mercaptoethylamine under stirring, stir for 24 hours, centrifuge at 13000 rpm for 8 minutes to obtain a precipitate, add ultrapure water to the precipitate Then, centrifuge at 13000rpm for 8min to obtain a precipitate, add ultrapure water to redissolve the precipitate, and repeat this 2~3 times until it is neutral, freeze-dry to obtain an aminated hollow mesoporous copper sulfide complex (CuS-NH ₂ );

(3) Synthesis of Fe3O4 nanoparticles: Dissolve 0.9 g FeCl ₂ and 2.4 g FeCl ₃ in 45 ml of water under stirring, heat to 80°C under nitrogen protection, slowly add 6 ml of ammonia water dropwise, stir in oil bath for 30 min, add mass 2.2ml of citric acid with a concentration of 0.5g·ml ^-1 was heated to 95°C, reacted for 90min, cooled to room temperature under stirring, dialyzed to remove unreacted substances, and freeze-dried to obtain water-soluble iron ferric oxide nanoparticles;

(4) Take 5.5 mg of the aminated hollow mesoporous copper sulfide complex obtained in step (2), disperse it in 5.5 ml of phosphate buffer solution with pH 7-9, ultrasonicate it for 10-30 min in an ice bath, and mix with the Tumor drugs are mixed with phosphate buffer, each 3ml of phosphate buffer contains 9g of anti-tumor drugs, stirred at room temperature for 24 hours, centrifuged and washed with water for 3 times, the probe is ultrasonically dispersed in water, freeze-dried to obtain the amination vulcanization of anti-tumor drugs copper;

(5) Disperse the aminated copper sulfide loaded with antineoplastic drugs in 22ml of water, add iron ferric oxide nanoparticles under stirring, stir at room temperature for 6-48h, centrifuge at 13000rpm for 8min to obtain a precipitate, and freeze-dry to obtain the loaded antineoplastic drug magnetic hollow mesoporous copper sulfide;

(6) Ultrasonically disperse the above drug-loaded magnetic hollow mesoporous copper sulfide in 22ml of water, add 220mg of hyaluronic acid in aqueous solution, stir and react for 6-48h, and then the magnetic hollow mesoporous copper sulfide modified by hyaluronic acid is obtained. pharmaceutical composition.

The application of the hyaluronic acid-modified magnetic hollow mesoporous copper sulfide pharmaceutical composition in the preparation of injection, oral or implantation drug for treating tumors.

The application of the hyaluronic acid-modified magnetic hollow mesoporous copper sulfide pharmaceutical composition in the preparation of anti-tumor drugs, the hyaluronic acid-modified magnetic hollow mesoporous copper sulfide pharmaceutical composition and pharmaceutically active or pharmacologically active molecules Combined to form a nanometer drug-carrying system, the pharmaceutically active or pharmacologically active molecules are antitumor drugs such as adriamycin, paclitaxel, docetaxel, hydroxycamptothecin, and mitoxantrone.

The application of the hyaluronic acid-modified magnetic hollow mesoporous copper sulfide pharmaceutical composition in the preparation of tumor photothermal therapy combined with chemotherapy, magnetic field remote control drug release, and magnetic resonance imaging drugs.

The hyaluronic acid-modified magnetic hollow mesoporous copper sulfide pharmaceutical composition of the present invention has good biocompatibility and low toxicity, and can transport the drug to the tumor site through the magnetic targeting of ferric iron tetroxide. Moreover, the drug-loading system can regulate the release of drugs under the action of an external magnetic field, and can realize the synergistic effect of hyperthermia and chemotherapy under laser irradiation, and effectively kill cancer cells. The prepared compound has multiple functions such as photothermal therapy, drug therapy, magnetic targeting, magnetic resonance imaging, etc., is convenient to operate, and the method is stable and reliable. The prepared copper sulfide and its pharmaceutical composition are biocompatible Good, strong magnetic targeting, less toxic and side effects, etc., in the treatment of tumors, it can play a role in hyperthermia, tumor diagnosis, magnetic field slow-controlled drug release, etc. The relevant information is as follows:

1. In vitro hyperthermia experiment of magnetic hollow mesoporous copper sulfide composition loaded with doxorubicin

Prepare a series of magnetic hollow mesoporous copper sulfide solutions loaded with doxorubicin, the copper sulfide concentrations are: 200ug/ml, 100ug/ml, 50ug/ml, 25ug/ml, 12.5ug/ml, 0ug/ml. The 808 nm near-infrared laser was used to irradiate with an energy density of 2W/cm2, and the temperature of the solution was measured at 0, 0.5, 1, 1.5, 2, ^2.5 , 3, 3.5, 4, 4.5, and 5 min, and the recorded results showed When the concentration of the preparation is 200ug/ml, 100ug/ml, 50ug/ml, 25ug/ml, 12.5ug/ml, 0ug/ml, the temperature rises by 54°C, 38.9°C, 28.5°C, 19.4°C, 11.2°C within 5 minutes respectively °C, 2.5 °C. The results show that the magnetic hollow mesoporous copper sulfide has excellent photothermal conversion effect, and its photothermal conversion effect is concentration-dependent.

2. Drug release experiment of magnetic hollow mesoporous copper sulfide composition loaded with doxorubicin

Take 2 copies of CuS-IONP-HA/DOX nanocomposites, respectively, the alternating magnetic field group and the non-alternating magnetic field group, disperse in 50ml pH7.4 phosphate buffer saline, and place them on a shaker (37°C, 100rpm) , take samples at 1h, 3h, 6h, 8h, 12h, and 14h respectively, each sampling volume is 0.5ml, and then add 0.5ml of release medium, each time point of the magnetic field group is placed under the alternating magnetic field for 30min, The parameters are: frequency 50kHz, magnetic field strength 2.5kA/m. The samples were detected by HPLC, and after 14 hours of drug release, the drug release of CuS-IONP-HA /DOX nanocomposite in the alternating magnetic field group was 41% higher than that in the non-alternating magnetic field group. The results show that in the presence of an alternating magnetic field, the ferroferric oxide in the CuS-IONP-HA/DOX composite can rotate and vibrate to deform, open the pores of the hollow mesoporous copper sulfide, and promote the drug release. .

3. Cell proliferation inhibition experiment of magnetic hollow mesoporous copper sulfide composition loaded with doxorubicin

Using the SRB method, select the MCF-7 human breast cancer cells in the logarithmic growth phase, adjust the cell number to 5×10 ⁴ /ml and inoculate them in a 96-well culture plate, 100 μl per well (the edge wells are filled with sterile PBS), and the cells are pasted. Drugs were added after 24 hours of wall growth, followed by blank group, CuS-IONP-HA group, DOX group, CuS-IONP-HA /DOX group, the final drug concentration was set at 4 μg/ml, and a laser group (2W/cm ² , 3min, 4h after dosing and laser irradiation) and the non-laser group, with 6 replicate wells for each group. After 24 h, add 50 μl of 50% trichloroacetic acid (TCA) pre-cooled at 4°C to each well to fix the cells, fix for 10 min, move to a 4°C refrigerator for 1 h, remove and discard the fixative, wash 5 times with deionized water, and shake dry. Dry naturally at room temperature. After drying at room temperature, add 50 μl of SRB staining solution to each well, place in the dark at room temperature for 15-30 minutes for staining, discard the staining solution, wash 5 times with 1% glacial acetic acid, and dry at room temperature. Afterwards, use 150 μl of unbuffered Tris lye (10mM, pH=10.5) to dissolve the dye bound to the cell protein, and after micro-oscillation on a shaker (37°C, 100rpm, 10min), measure each small well at a wavelength of 515nm on a microplate reader OD value of the tumor cell growth inhibition rate (%)=(1-OD value of the experimental group/OD value of the control group)×100% was calculated to calculate the CuS-IONP-HA group, CuS-IONP-HA -laser group, DOX group, DOX-laser group, CuS-IONP-HA /DOX group, and CuS-IONP-HA /DOX-laser group, the cell growth inhibition rates of each group were: 4.9%, 25.8%, 49.7%, 53.3%, 51.2% , 77.4%. The results show that the carrier CuS-IONP has no obvious toxicity to cells at the experimental dose, but has obvious photothermal effect under the irradiation of near-infrared light (808nm); the cytotoxicity difference between the preparation group and the raw material drug group when there is no near-infrared light irradiation The inhibitory effect of CuS-IONP-HA/DOX on tumor cells was the strongest when irradiated by near-infrared light, which was the result of the synergistic treatment of chemotherapy and hyperthermia.

4. Pharmacodynamic study of magnetic hollow mesoporous copper sulfide composition loaded with doxorubicin

Purchase Kunming mice (female, 3-4 weeks old), subcutaneously inoculate S-180 ascites tumor cells on the back of the right upper limb of the mouse, measure the tumor volume 7 days later, and take the tumor volume ≥ 100 mm ³ and the tumor volume and body weight are similar Mice were randomly divided into 10 groups, 6 in each group. The specific groups are as follows: normal saline group, normal saline-laser group, CuS-IONP-HA group, CuS-IONP-HA-laser group, CuS-IONP-HA -laser/magnet group, DOX group, DOX-laser group, CuS -IONP-HA /DOX group, CuS-IONP-HA/DOX-laser group, CuS-IONP-HA/DOX-laser/magnet group, the light source used in the laser group is 808nm near-infrared laser with a power of 2W/cm ² , 3 hours after the administration, the laser was irradiated to the tumor site, and the irradiation time was 1 minute each time. The mice in each group were administered by tail vein injection, once every two days, for a total of 7 times. During the whole experiment, the mice were kept on a normal diet every day, the weight of each mouse was weighed every two days, and the long diameter (A) and short diameter (B) of the sarcoma of the tumor-bearing mice were measured with a digital display vernier caliper, according to the formula Tumor volume V=A×B ² /2 Calculate the tumor volume. The recorded data showed that CuS-IONP-HA group, CuS-IONP-HA-laser group, CuS-IONP-HA-laser/magnet group, DOX group, DOX-laser group, CuS-IONP-HA/DOX group, CuS -The tumor inhibition rates of IONP-HA/DOX-laser group and CuS-IONP-HA/DOX-laser/magnet group were 4.49%, 26.82%, 41.56%, 43.85%, 46.03%, 68.75%, 81.43%, 87.68% %. The results showed that the drug effect of the CuS-IONP-HA/DOX-laser/magnet group was significant. When the administration of magnetic hollow mesoporous copper sulfide and its pharmaceutical composition was combined with laser irradiation and an external magnetic field, the tumor volume of the mice was increased. It shows that the combination of CuS-IONP-HA/DOX-laser/magnet tumor-targeted drug delivery system chemotherapy, 808nm laser hyperthermia, and magnetic targeting can significantly enhance the therapeutic effect of tumors.

5. Magnetic resonance imaging experiment of magnetic hollow mesoporous copper sulfide composition loaded with doxorubicin

S-180 ascites tumor cells were inoculated subcutaneously on the back of the right upper limb of the mice, and the tumor volume was measured 7 days later. 12 mice with tumor volume ≥ 100 mm ³ and similar tumor volume and body weight were randomly divided into two groups, each There were 6 rats in the group, and the specific groups were as follows: normal saline group, CuS-IONP-HA/DOX group. The two groups of mice were anesthetized by intraperitoneal injection of 0.04ml 3% pentobarbital sodium. After fixation, the mice of the two groups were administered intravenously, including 200 μl of normal saline and 2 mg/ml of CuS-IONP-HA/DOX Physiological saline solution 200 μl. T2WI magnetic resonance imaging was performed on mice 24 hours after injection. The scanning parameters are: SE-T2WI at transverse position, TR4240ms, TE1108.47ms, FOV8cm, slice thickness (2mm at transverse position, 3mm at coronal position), slice spacing 1mm, matrix 256*256. On the T2WI images of the tumor areas of the two groups of mice, a region of interest (ROI) of the same size was drawn to measure the signal intensity (SIT) of the solid part of the tumor, and the average value was taken, and the measurement area was not less than 6mm ² . The results showed that the T2WI signal of the tumor area in the CuS-IONP-HA/DOX group was significantly lower than that of the corresponding area in the control group, and the effect of MRI was significantly enhanced.

Experiments show that, compared with the prior art, the present invention has the following outstanding beneficial technical effects:

(1) The magnetic hollow mesoporous copper sulfide and its pharmaceutical composition provided by the present invention have excellent biocompatibility, water dispersibility and stability, and retain the high-efficiency photothermal therapy activity of copper sulfide, which can collect heat and Chemotherapy as one;

(2) The present invention chooses ferroferric oxide to modify copper sulfide, which can not only provide magnetic targeting in the process of drug transport, but also act as a capping agent to prevent drug leakage, and under the action of an external magnetic field, it can prevent drug leakage. The release plays the role of controlled release. At the same time, ferric oxide can be used for magnetic resonance imaging, and real-time monitoring can be carried out during the treatment process, which provides a new direction for the diagnosis and treatment of tumors.

Claims (4)

1. A kind of 1. preparation method of the pharmaceutical composition of the mesoporous copper sulfide of hyaluronic acid decorated magnetic hollow, it is characterised in that Antineoplastic, sulphur are loaded on the amidized copper sulfide that hollow mesoporous copper sulphide nano particles and mercaptoethylmaine react to obtain Change the upper ferriferrous oxide nano grain of copper surface electrostatic absorption, through amido link reaction hyaluronic acid in surface modification, into hyalomitome The pharmaceutical composition of the mesoporous copper sulfide of magnetic hollow of acid modification；Described ferroso-ferric oxide and the mass content ratio of copper sulfide are 1 ~ 8 ︰ 1, the particle diameter of the mesoporous copper sulfide of magnetic hollow is 50 ~ 300nm, and preparation method comprises the following steps：

   （1）, by 0.7-0.8g CuSO₄·5H₂O is dissolved in 180-220ml water, stirring, adds 5-7g polyvinylpyrrolidines Ketone, 10 ~ 20min is reacted, add 1.1-1.3g sodium hydroxides, stir, then 11-13ml hydrazine hydrates are added dropwise, stirring 5 ~ 10min, 1.2-1.4ml ammonium sulfate is added, 1h is stirred, with ultra-pure water 12000-15000rpm（That is 12000-15000r/min）From Heart 5-10min is washed to neutrality, is freeze-dried 48h, obtains hollow mesoporous nano copper sulfate particle；

   （2）, by hollow mesoporous copper sulfide nano grain of rice 90-110mg ultrasonic disperses in 90-110ml ultra-pure waters, ultrasound 10 ~ 30min, lower addition 180-220mg mercaptoethylmaines are stirred, stir 24h, 12000-15000rpm centrifugations 5-10min must be precipitated, sunk Forming sediment plus ultra-pure water redissolves, then must be precipitated in 12000-15000rpm centrifugations 5-10min, precipitation plus ultra-pure water redissolve, and so repeatedly 2 ~ 3 times, until being neutrality, freeze-drying, obtain the hollow mesoporous vulcanization copper composition (CuS-NH of amination₂)；

   （3）, synthesis ferriferrous oxide nano grain：Take 0.8-0.9 g FeCl₂With 2.3-2.4 g FeCl₃35- is dissolved under stirring In 45ml water, 80 DEG C are heated under nitrogen protection, 4-6ml ammoniacal liquor is slowly added dropwise, oil bath stirring 30min, adds mass concentration 0.5g·ml^-1Citric acid 1.8-2.2ml, be warming up to 95 DEG C, react 90min, room temperature is cooled under stirring, dialysis removes not anti- Material is answered, is freeze-dried, produces water-soluble ferroferric oxide nanoparticle；

   （4）, take step（2）The obtained hollow mesoporous vulcanization copper composition (CuS-NH of amination₂) 4.5-5.5mg, it is scattered in In pH7-9 phosphate buffer 4.5-5.5ml, 10 ~ 30 min of ultrasound, delay with the phosphate containing antineoplastic under ice bath Fliud flushing mixes, and contains 9g antineoplastics in every 3ml phosphate buffers, and 24h is stirred at room temperature, and centrifugation is washed with water 3 times, pops one's head in Ultrasonic disperse is freeze-dried the amination copper sulfide for producing load antineoplastic in water；

   （5）, the amination copper sulfide for loading antineoplastic is scattered in 18-22ml water, stir and lower add ferroso-ferric oxide Nanoparticle, is stirred at room temperature 6-48h, and 12000-15000rpm centrifugations 5-10min must be precipitated, is freeze-dried, it is antitumor produce load The mesoporous copper sulfide of magnetic hollow of medicine；

   （6）, by the mesoporous copper sulfide ultrasonic disperse of the magnetic hollow of above-mentioned carrying medicament in 18-22ml water, add 180-220mg Hyaluronic acid aqueous solution, stirring reaction 6-48h, the hyaluronic acid decorated mesoporous copper sulfide of magnetic hollow drug regimen Thing.
2. 2. the preparation side of the pharmaceutical composition of the mesoporous copper sulfide of hyaluronic acid decorated magnetic hollow according to claim 1 Method, it is characterised in that comprise the following steps：

   （1）, by 0.75g CuSO₄·5H₂O is dissolved in 200ml water, stirring, adds 6.0g polyvinylpyrrolidones, reaction 10 ~ 20min, 1.2g sodium hydroxides are added, after stirring, 12ml hydrazine hydrates are added dropwise with syringe, stir 5 ~ 10min, finally add Enter 1.29ml ammonium sulfate, stir 1h, be washed to neutrality with ultra-pure water 15000rpm centrifugations 5-10min, be freeze-dried 48h, obtain Hollow mesoporous nano copper sulfate particle；

   （2）, hollow mesoporous copper sulfide nano grain of rice 100mg Probe Ultrasonic Searchings are scattered in 100ml ultra-pure waters, 10 ~ 30min of ultrasound, 200mg mercaptoethylmaines are added under stirring condition, stir 24h, 15000rpm centrifugations 5-10min must be precipitated, and add ultra-pure water to redissolve, then Centrifugation, so repeatedly 2 ~ 3 times, until being neutrality, freeze-drying, obtain the hollow mesoporous vulcanization copper composition (CuS- of amination NH₂)；

   （3）, synthesis ferriferrous oxide nano grain：Take 0.86 g FeCl₂With 2.35 g FeCl₃It is dissolved under stirring in 40ml water, 80 DEG C are heated under nitrogen protection, 5ml ammoniacal liquor is slowly added dropwise with syringe, oil bath stirring 30min, adds mass concentration afterwards 0.5g·ml^-1Citric acid 2ml, be warming up to 95 DEG C, react 90min, room temperature is cooled under stirring, dialysis removes unreacted reactant Matter, freeze-drying, produces water-soluble ferroferric oxide nanoparticle；

   （4）, take the hollow mesoporous vulcanization copper composition 5mg of amination, be scattered in pH7-9 5ml phosphate buffers, under ice bath The min of Probe Ultrasonic Searching 10 ~ 30, is mixed with the phosphate buffer containing antineoplastic, is contained in every 3ml phosphate buffers 9g antineoplastics, 24h is stirred at room temperature, 15000rpm centrifugations 5-10min is washed with water 3 times, and Probe Ultrasonic Searching is dispersed in water, cold Freeze the amination copper sulfide for being drying to obtain load antineoplastic；

   （5）, by load antineoplastic amination copper sulfide be scattered in 20ml water, under stirring condition add be dispersed in water Ferriferrous oxide nano grain, 24h is stirred at room temperature, centrifuging to precipitate, freeze-drying, produce load antineoplastic magnetic in Empty mesoporous copper sulfide；

   （6）, the mesoporous copper sulfide Probe Ultrasonic Searching of the magnetic hollow of carrying medicament is scattered in 20ml water, add 200mg hyalomitomes Aqueous acid, stirring reaction 24h, produce the pharmaceutical composition of the mesoporous copper sulfide of hyaluronic acid decorated magnetic hollow.
3. 3. the pharmaceutical composition of the mesoporous copper sulfide of hyaluronic acid decorated magnetic hollow prepared by the methods described of claim 1 or 2 Application in antineoplastic composition injection, oral agents or drug delivery implant agent is prepared, adriamycin, the purple of described antineoplastic China fir alcohol, Docetaxel, HCPT, mitoxantrone.
4. 4. the pharmaceutical composition of the mesoporous copper sulfide of hyaluronic acid decorated magnetic hollow prepared by the methods described of claim 1 or 2 Application in preparing tumor thermal therapy combined chemotherapy, magnetic field and regulating and controlling drug release, magnetic resonance imaging medicine at a distance.