Acoustic dynamic targeting nano material as well as preparation method and application thereof

Abstract

The invention discloses a sonodynamic targeting nano material as well as a preparation method and application thereof, and belongs to the technical field of medicines. The nano material comprises a chemotherapeutic drug, a thermogenic sound-sensitive agent, a metal organic framework entrapping the chemotherapeutic drug and the thermogenic sound-sensitive agent, a liposome membrane entrapping the metal organic framework, and a tumor marker loaded on the surface of the liposome membrane. After entering a human body, the nano-material can be combined with tumor cells in a targeted manner, a sound-sensitive agent in the nano-material is excited by sonodynamic force to generate heat and a chemotherapeutic drug (such as paclitaxel) is released, so that a thermochemotherapy process in tumor peritoneal metastasis treatment is simulated, and thermotherapy and chemotherapy are combined to obtain a new tumor peritoneal metastasis treatment scheme; according to the treatment scheme, side effects of the whole body of a patient are reduced while the chemotherapy effect is synergistically improved, the treatment effect of the patient is improved, and the life cycle of the patient is prolonged.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to a sonodynamic targeting nanomaterial, a preparation method thereof, and an application thereof. Background Art

[0002] The information disclosed in the background art of the present invention is only intended to increase the understanding of the overall background of the present invention, and does not necessarily be regarded as an admission or imply in any form that this information constitutes the prior art already known to those of ordinary skill in the art.

[0003] Distant metastasis of tumors is the main cause of death in cancer patients, including lung metastasis, liver metastasis, brain metastasis, bone metastasis, etc. Among them, peritoneal metastasis is a frequently overlooked tumor metastasis pattern. For malignant tumors prone to peritoneal metastasis (such as ovarian cancer, colorectal cancer, gastric cancer, cholangiocarcinoma, etc.), peritoneal metastasis is often the most important prognostic factor, which can lead to problems such as low radical resection rate of surgery, chemotherapy insensitivity, and easy recurrence of tumors, greatly reducing the survival time and quality of life of cancer patients.

[0004] In the early 20th century, studies have found that early chemotherapy and palliative cytoreductive surgery can temporarily improve the symptoms of patients with peritoneal metastatic tumors, but it is difficult to continuously control tumor progression and extend the prognosis. With the continuous progress of systemic treatment means, the optimization of cytoreductive surgery techniques, and the combination with local treatment (especially hyperthermic intraperitoneal chemotherapy), patients with peritoneal metastatic tumors have also obtained more effective multimodal treatment options. Hyperthermic intraperitoneal chemotherapy (HIPEC) is to fill the abdominal cavity with a perfusion fluid containing chemotherapy drugs, precisely keep the temperature constant and circulate at a constant speed for a certain period of time, combine intraperitoneal chemotherapy and hyperthermia, and through the direct killing effect of high temperature on tumors, the synergistic effect of high temperature and chemotherapy drugs in anti-tumor, and the mechanical flushing effect, to achieve the therapeutic effect of clearing free cancer cells, subclinical lesions, and microcarcinomas in the abdominal cavity, thereby playing a preventive and therapeutic role in cancer cell peritoneal metastasis.

[0005] However, for patients in the advanced stage of cancer, due to their poor tolerance and many complications, the actual clinical treatment effect of the existing hyperthermic intraperitoneal chemotherapy is not good. Sonodynamic therapy (SDT) of tumors is a treatment method developed on the basis of photodynamic therapy. In this method, after the tumor absorbs a sonosensitizer (chemical), a certain intensity of ultrasonic wave (physical) is applied to irradiate the tumor. The sonosensitizer generates cytotoxic reactive oxygen species (singlet oxygen O2, free radicals, etc.) through sonochemical reactions, causing irreversible damage to tumor cells and resulting in the death of tumor cells, realizing non-surgical targeted treatment of tumors.

[0006] Therefore, a new technical solution is needed to reduce systemic side effects while synergistically improving the chemotherapy effect, ultimately aiming to improve the treatment effect of cancer peritoneal metastasis, enhance the treatment effect of patients, and extend the life cycle. Summary of the Invention

[0007] Aiming at the deficiencies in the prior art, the purpose of the present invention is to provide a sono - dynamic targeted nanomaterial. After entering the human body, this nanomaterial can generate heat and release chemotherapy drugs through sono - dynamic excitation of the material, combining hyperthermia and chemotherapy to synergistically improve the chemotherapy effect while reducing systemic side effects.

[0008] To achieve the above - mentioned purpose, the present invention adopts the following technical solutions:

[0009] A sono - dynamic targeted nanomaterial includes a chemotherapy drug, a heat - generating sonosensitizer, a metal - organic framework encapsulating the chemotherapy drug and the heat - generating sonosensitizer, a liposome membrane coating the metal - organic framework, and a tumor marker loaded on the surface of the liposome membrane. After entering the human body, this nanomaterial can target and bind to tumor cells, generate heat through sono - dynamic excitation of the sonosensitizer in the nanomaterial and release chemotherapy drugs (such as paclitaxel), so as to simulate the thermo - chemotherapy process in the treatment of tumor peritoneal metastasis.

[0010] Preferably, the chemotherapy drug is any one of paclitaxel, docetaxel, camptothecin, 5 - fluorouracil, cisplatin, oxaliplatin, irinotecan, doxorubicin, mitomycin, epirubicin, docetaxel, oxaliplatin or irinotecan.

[0011] More preferably, the chemotherapy drug is paclitaxel, a commonly used chemotherapy drug for peritoneal metastasis by intraperitoneal perfusion chemotherapy.

[0012] Preferably, the metal - organic framework is ZIF - 8. ZIF - 8 has excellent properties such as a high specific surface area, regular pore size, thermal stability, water stability, and low toxicity, and has pH sensitivity, being more likely to release in the slightly acidic tumor microenvironment. Therefore, it can be used as a tumor drug carrier.

[0013] Preferably, the heat - generating sonosensitizer is Ce6.

[0014] Preferably, the tumor marker is selected from any one of carcinoembryonic antigen - like markers, enzyme - like markers, hormone - like markers, glycoprotein - like markers, and oncogene - like markers.

[0015] More preferably, the tumor marker is EpCAM antibody.

[0016] The present invention also provides a method for preparing a sonodynamic targeting nanomaterial, which comprises the following steps: blending a metal-organic framework, a chemotherapeutic drug, and a thermogenic sonosensitizer in an organic solvent, fully reacting to obtain a precipitate, separating and purifying the precipitate to obtain a metal cluster-based group; dispersing and dissolving the metal cluster-based group and a liposome membrane loaded with a tumor marker in PBS, and performing ultrasonic emulsification under an ice bath to prepare the sonodynamic targeting nanomaterial.

[0017] Further, the liposome membrane loaded with the tumor marker is prepared by the thin film dispersion method.

[0018] Another object of the present invention is to provide the application of the sonodynamic targeting nanomaterial as described above in the preparation of a tumor peritoneal metastasis hyperthermia system, and the hyperthermia system includes:

[0019] An injection module, the injection module includes a liquid storage unit and an intraperitoneal injection unit, and the liquid storage unit stores a hyperthermia solution prepared from the above-mentioned sonodynamic targeting nanomaterial;

[0020] An ultrasonic module;

[0021] An observation module, the observation module includes an intraperitoneal infrared imaging unit and a display unit.

[0022] The beneficial effects of the present invention are as follows:

[0023] The present invention provides a sonodynamic targeting nanomaterial and its application. After the nanomaterial enters the human body, it can target and bind to tumor cells, and generate heat through sonodynamic excitation of the sonosensitizer in the nanomaterial and release chemotherapeutic drugs (such as paclitaxel), so as to simulate the hyperthermia chemotherapy process in the treatment of tumor peritoneal metastasis, combine hyperthermia with chemotherapy, and obtain a new treatment plan for tumor peritoneal metastasis; while synergistically improving the chemotherapy effect, this treatment plan reduces the systemic side effects of patients, improves the treatment effect of patients, and prolongs the life cycle of patients.

[0024] The above description is only an overview of the technical solution of the present invention. In order to be able to understand the technical means of the present invention more clearly and implement it according to the content of the specification, the following will take the preferred embodiments of the present invention and cooperate with the drawings to describe in detail as follows. Description of the Drawings

[0025] 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 to be used in the description of the specific embodiments or the prior art. Obviously, some of the following drawings are embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained according to these drawings.

[0026] Figure 1Characterization of PCM@LIP-Ep: A. TEM image of ZIF-8; B. TEM image of PTX / Ce6-MOF@LIP-EP (PCM@LIP-Ep); C. SEM image of PTX / Ce6-MOF and elemental MAPPING (scale bar = 100 nm);

[0027] Figure 2 Results of PCM@LIP-Ep in treating peritoneal metastasis of mouse tumors. Detailed implementation manners

[0028] To better describe the present invention, the following specific examples are given for further illustration. The methods in the following examples are all conventional methods unless otherwise specified.

[0029] The technical solutions of the present invention are all conventional solutions in the art unless otherwise specified; the reagents or materials are all from commercial channels unless otherwise specified.

[0030] The following detailed descriptions are all illustrative and are intended to provide further explanations for the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present application belongs. The following examples are only for explaining the present invention and do not limit its content. If the specific experimental conditions are not indicated in the examples, they are usually in accordance with conventional conditions or the conditions recommended by reagent companies; the reagents, consumables, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.

[0031] The abbreviations used in the following examples are as follows:

[0032] Example 1 Preparation of the nanomaterial PCM@LIP-Ep

[0033] 1) Preparation of PTX-Ce6-MOF (PCM): At room temperature, under magnetic stirring, 30 mmol of Zn(NO3)2·6H2O and 200 mmol of 2-IMM were respectively dissolved in 300 ml and 200 ml of methanol, and 50 mg of ce6 and 50 mg of PTX (dissolved in DMSO) were added. Stir at room temperature for 12 hours. The precipitate was centrifuged and washed three times with methanol to remove residual precursors, and then freeze-dried to obtain PCM.

[0034] 2) Preparation of liposomes by thin film dispersion method: Weigh 100 mg of dioleoyl phosphatidylcholine (DOPC), 5 mg of dioleoyl phosphatidylethanolamine (DOPE), 5 mg of DSPE-mPEG2000-EpCAM antibody, and 30 mg of cholesterol into a round-bottom flask, and add 10 mL of chloroform to dissolve them completely. Connect the round-bottom flask to a rotary evaporator, and adjust the parameters: rotation speed 60 rpm, heating condition 40 °C, running time 15 min. After a lipid film appears at the bottom of the round-bottom flask, stop the vacuum, resuspend the film with 4 mL of PBS and disperse it by ultrasound to obtain a relatively homogeneous liposome-PBS dispersion system.

[0035] 3) Weigh an appropriate amount of PCM and wash it several times with PBS, and then redisperse it with fresh PBS. Add the dispersed metal cluster-based MOF and liposomes into a 10 mL centrifuge tube, and perform ultrasonic emulsification in an ice bath. Relevant parameters: ultrasonic time 2 s, intermittent time 2 s, ultrasonic power 325 w, ultrasonic time 3 minutes. Place the MOF@NPs-modified nanoparticles obtained after ultrasound at 4 °C for storage.

[0036] Physicochemical properties of PCM@LIP-Ep nanomaterials:

[0037] Please refer to Figure 1 , according to the transmission electron microscope TEM ( Figure 1 A) The results show that ZnMOF (ZIF-8) has a uniform structure. After encapsulating Ce6-PTX and covering the liposome membrane, PTX-Ce6@ZnMOF@LIP-EpCAM (PCM@LIP-Ep) was constructed, with a particle size of about 200 nm, and the outer membrane structure could be identified under the microscope ( Figure 1 B). The TEM elemental mapping indicates that C, N, Zn, and O elements are uniformly distributed in the material ( Figure 1 C).

[0038] Example 2 Thermal chemotherapy effect of the nanomaterial PCM@LIP-Ep obtained in Example 1 in the abdominal cavity

[0039] 2.1 Experimental animals: Male nude mice, 6 - 8 weeks old, weighing 18 - 25 g. Provided by Lingchang Company, and fed adaptively for one week before the experiment.

[0040] 2.2 Experimental drugs

[0041] The PCM@LIP-Ep nanoparticles obtained in Example 1 (redisperse the nano PCM@LIP-Ep colloidal solution with 0.9% NaCl injection solution, physical stability > 8 hours); Ce6 nanoparticles (redisperse the nano PCM@LIP-Ep colloidal solution with 0.9% NaCl injection solution, physical stability > 8 hours); PBS solution.

[0042] 2.3 Experimental steps:

[0043] The mice in the experimental group were intraperitoneally injected with PCM@LIP-Ep (10 μg / 1 g body weight of mice); the mice in control group 1 were intraperitoneally injected with Ce6 (5 μg / 1 g body weight of mice); the mice in control group 2 were intraperitoneally injected with PBS solution.

[0044] 12 h after the intraperitoneal injection, each group of mice was treated with ultrasound for 5 min.

[0045] 2.4 Experimental results:

[0046] Please refer to Figure 2 , during the ultrasound treatment, the infrared imaging results showed that the temperature in the abdomen of the mice in the experimental group injected with PCM@LIP-EP increased significantly during the ultrasound treatment, reaching up to 46 °C, and the heating area was concentrated in the tumor site, proving that it played a role in thermochemotherapy in the abdomen.

[0047] In summary, the present invention provides a sonodynamic targeting nanomaterial and its application. After entering the human body, the nanomaterial can target and bind to tumor cells, and through sonodynamic excitation, the photosensitizer in the nanomaterial generates heat and releases chemotherapeutic drugs (such as paclitaxel) to simulate the thermochemotherapy process in the treatment of tumor peritoneal metastasis, combining hyperthermia and chemotherapy to obtain a new treatment plan for tumor peritoneal metastasis; while synergistically improving the chemotherapy effect, this treatment plan reduces the systemic side effects of patients, improves the treatment effect of patients, and prolongs the life cycle of patients.

[0048] The technical features of the above-described embodiments can be combined arbitrarily. For the sake of concise description, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered as the scope described in this specification.

[0049] The above-described embodiments only represent several implementation manners of the present invention. Their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the invention patent should be subject to the appended claims.

Claims

1. A sono-dynamic targeting nanomaterial, characterized in that, It includes a chemotherapeutic drug, a thermogenic sonosensitizer, a metal-organic framework encapsulating the chemotherapeutic drug and the thermogenic sonosensitizer, a liposome membrane coating the metal-organic framework, and a tumor marker loaded on the surface of the liposome membrane.

2. The sono-dynamic targeting nanomaterial according to claim 1, characterized in that, The chemotherapeutic drug includes any one of paclitaxel, docetaxel, camptothecin, 5-fluorouracil, cisplatin, oxaliplatin, irinotecan, doxorubicin, mitomycin, epirubicin, docetaxel, oxaliplatin, irinotecan.

3. The sono-dynamic targeting nanomaterial according to claim 2, wherein The chemotherapeutic drug is paclitaxel.

4. The sono-kinetic targeting nanomaterial according to claim 1, characterized in that, The metal-organic framework is ZIF-8.

5. The sono-dynamic targeting nanomaterial according to claim 1, wherein The thermogenic sonosensitizer is Ce6.

6. The sono-dynamic targeting nanomaterial according to claim 1, wherein The tumor marker is selected from any one of carcinoembryonic antigen-like markers, enzyme-like markers, hormone-like markers, glycoprotein-like markers, oncogene-like markers.

7. The sono-dynamic targeting nanomaterial according to claim 1, wherein The tumor marker is EpCAM antibody.

8. A preparation method of the sono-dynamic targeting nanomaterial according to any one of claims 1 to 7, characterized in that, It includes the following steps: blending the metal-organic framework, the chemotherapeutic drug and the thermogenic sonosensitizer in an organic solvent, fully reacting to obtain a precipitate, separating and purifying the precipitate to obtain a metal cluster; dispersing and dissolving the metal cluster and the liposome membrane loaded with the tumor marker in PBS, and performing ultrasonic emulsification under ice bath to prepare a sonodynamic targeted nanomaterial.

9. The preparation method of the sono-dynamic targeting nanomaterial according to claim 8, characterized in that, The liposome membrane loaded with the tumor marker is prepared by the thin film dispersion method.

10. Use of a sono - dynamic targeting nanomaterial as described in any one of claims 1 - 7 in the preparation of a hyperthermia system for tumor peritoneal metastasis, characterized in that, The hyperthermia system includes: An injection module, the injection module includes a liquid storage unit and an intraperitoneal injection unit, and the liquid storage unit stores a hyperthermia solution configured with the sonodynamic targeted nanomaterial according to any one of claims 1 to 7; An ultrasonic module; An observation module, the observation module includes an intraperitoneal infrared imaging unit and a display unit.

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