Application of nano-drug and combined radiotherapy in cancer treatment

Abstract

The invention discloses an application of a nano-drug and combined radiotherapy in cancer treatment, and relates to the technical field of nanotechnology and biomedicine. The nano-drug (GL) obtained by the simple preparation method is a gambogic acid loaded nano-liposome, and the GL has good biocompatibility and passive targeting property. According to the invention, GL and FLASH radiotherapy are combined to treat cancers, and through the synergistic effect of GL and FLASH radiotherapy, double striking on tumor cells is realized, so that the anti-tumor effect is remarkably enhanced, side effects in the treatment process are reduced, and the life quality of patients is improved. Besides, the combined treatment strategy effectively overcomes the problem that cancer is not sensitive to existing radiotherapy and chemotherapy, more remarkable and efficient anti-tumor effect is achieved, more treatment choices and hopes are brought to cancer patients, and a new way is opened up for cancer treatment.

Description

Technical Field

[0001] The present invention relates to the field of nanotechnology and biomedical technology, and in particular to an application of nanomedicine and combined radiotherapy in cancer treatment. Background Art

[0002] Garcinia acid (GA) is a caged xanthone small molecule compound derived from the traditional Chinese medicine Garcinia. 38 H 44 O8. Modern pharmacological studies have shown that garcinia acid has multiple therapeutic activities such as anti-inflammatory and anti-oxidative, anti-tumor, improvement of retinopathy, organ protection, anti-microbial infection, relief of neuropathic pain and bone protection. In recent years, the therapeutic scope of garcinia acid has been continuously broadened, and more pharmacological effects have been disclosed, fully demonstrating its great prospects in disease treatment. Since garcinia acid has extremely low solubility in water (less than 0.5 μg / mL), its poor water solubility and poor stability lead to low bioavailability of garcinia acid, which greatly limits its clinical use.

[0003] Ultra-high dose rate radiotherapy, also known as flash radiotherapy, FLASH radiotherapy, flash therapy or ultra-high dose rate radiotherapy, is an emerging treatment technology that uses ultra-high dose rate and ultra-fast radiation to produce the FLASH effect that has a protective effect on normal tissues. FLASH radiotherapy delivers ultra-high doses of radiation at ultra-high dose rates (approximately ≥40Gy / s), which significantly broadens the treatment window for patients and significantly changes the pattern of radiotherapy and tumor treatment. Compared with conventional dose rate (1-7Gy / s) radiotherapy, FLASH radiotherapy can deliver an irradiation dose of more than 8Gy in a very short time, with a dose rate of more than 40Gy / s. Therefore, compared with conventional radiotherapy, FLASH radiotherapy, with its unique radiobiological advantages and ultra-short treatment time, can deliver large doses of irradiation to the tumor site, which is very likely to bring a new revolution to the radiotherapy of clinical malignant tumors.

[0004] However, both GA-related drug-guided chemotherapy and FLASH radiotherapy alone have tolerability issues, limiting their widespread application in cancer treatment.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] The purpose of the present invention is to provide an application of nanomedicine and combined radiotherapy in cancer treatment to solve the above technical problems.

[0007] The present invention is achieved in that:

[0008] In a first aspect, an embodiment of the present invention provides an application of nanomedicine combined with radiotherapy in cancer treatment, wherein the nanomedicine (GL) is a nanoliposome loaded with gambogic acid, and the radiotherapy method is selected from any one of photon-ultra-high dose rate radiotherapy (X-FLASH) and electron-ultra-high dose rate radiotherapy (E-FLASH).

[0009] In a second aspect, an embodiment of the present invention provides a nanomedicine as described above, wherein the nanomedicine comprises gambogic acid, cholesterol and lecithin in a mass ratio of 1:(1.5-2.5):(14-15).

[0010] The present invention has the following beneficial effects:

[0011] The nanomedicine (GL) provided in the embodiment of the present invention, i.e., the nanoliposome loaded with gambogic acid, is an innovative nanomedicine. The gambogic acid is encapsulated in the nanoliposome to form a stable and uniform GL solution. This nanoliposome has good biocompatibility and passive targeting, can efficiently deliver gambogic acid to the tumor site, reduce the distribution of the drug in normal tissues, thereby improving the anti-tumor effect of the drug and reducing side effects. In addition, by making liposomes, the shortcomings of poor water solubility and poor stability of gambogic acid can be significantly improved, so that it can better play an anti-tumor effect.

[0012] The embodiments of the present invention improve the passive targeting of liposomes loaded with gambogic acid and the selectivity of FLASH radiotherapy, and through the synergistic effect of the two, a double blow to tumor cells is achieved, which not only significantly enhances the anti-tumor effect, but also reduces the side effects during the treatment process and improves the quality of life of patients. The combined treatment strategy of the present invention effectively overcomes the problem that cancer is insensitive to existing radiotherapy and chemotherapy, achieves a more significant and efficient anti-tumor effect, brings more treatment options and hope to cancer patients, and opens up new ways for cancer treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0014] Figure 1 The appearance and state of the GL solution in Example 1;

[0015] Figure 2 is the microscopic morphology of GL in Example 1;

[0016] Figure 3Tumor inhibition rate for X-FLASH radiation dose optimization process;

[0017] Figure 4 Figure 2 shows the growth curve of mouse tumor volume during combined treatment of pancreatic cancer (a) and the curve of mouse body weight change (b).

[0018] Figure 5 The tumor morphology of mice after combined treatment of pancreatic cancer (a) and the tumor volume of mice (b);

[0019] Figure 6 The tumor mass of mice after combined treatment of pancreatic cancer (a) and the tumor inhibition rate of mice (b);

[0020] Figure 7 Results of the detection of physiological indicators in mouse serum after the combined treatment of pancreatic cancer: TNF-α level (a); mouse serum IL-1β level (b); mouse serum IFN-γ level (c); mouse serum IL-10 level (d); mouse serum TGF-β1 level (e);

[0021] Figure 8 The results of blood routine tests of mice after combined treatment of pancreatic cancer, from left to right: (a) percentage of white blood cells, neutrophils, lymphocytes, and monocytes; (b) hemoglobin and mean corpuscular hemoglobin concentration; (c) red blood cells and platelets;

[0022] Fig. 9 Results of blood biochemical tests of mice after combined treatment of pancreatic cancer: (a) albumin; (b) alanine aminotransferase; (c) aspartate aminotransferase; (d) creatinine; (e) serum total cholesterol; (f) triglycerides; (g) serum total protein; (h) urea nitrogen content;

[0023] Fig.10 H&E staining of the main organ tissues of mice after the combined treatment of pancreatic cancer;

[0024] Fig.11 The growth curve of tumor volume (a) and the curve of body weight change (b) of mice during combined treatment of breast cancer;

[0025] Fig.12 The tumor volume (a) and tumor mass (b) of mice after combined treatment of breast cancer;

[0026] Fig.13 Results of testing of physiological indicators in mouse serum after combined treatment of breast cancer: TNF-α level (a); mouse serum IL-1β level (b); mouse serum IFN-γ level (c); mouse serum IL-10 level (d); mouse serum TGF-β1 level (e). DETAILED DESCRIPTION

[0027] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0028] In a first aspect, an embodiment of the present invention provides an application of nanomedicine combined with radiotherapy in cancer treatment, wherein the nanomedicine (GL) is a nanoliposome loaded with gambogic acid, and the radiotherapy method is selected from any one of photon-ultra-high dose rate radiotherapy (X-FLASH) and electron-ultra-high dose rate radiotherapy (E-FLASH).

[0029] It should be noted that the main radiotherapy techniques currently include photon radiotherapy, electron radiotherapy, proton radiotherapy and heavy ion radiotherapy. The specific choice of radiotherapy technique depends on the type, location, size of the tumor, and the overall health of the patient. In the optional implementation of the present invention, any form of photon radiotherapy or electron radiotherapy is selected according to actual needs.

[0030] Specifically, photon radiotherapy uses X-rays or gamma rays, which can penetrate deep into the human body. The dose distribution is initially high-dose, then gradually decreases, forming a dose peak and then decreasing again. Photons indirectly damage cell DNA by generating free radicals, which can react with DNA, causing DNA chain breaks or mutations, thereby inhibiting the proliferation of tumor cells. Photon radiotherapy is suitable for most types of cancer, especially deep tumors, and is the most widely used technology in radiotherapy.

[0031] Electron radiotherapy has low energy and limited penetration, and is suitable for treating tumors on the skin or close to the body surface. The dose distribution of electron beams is characterized by a low dose near the skin surface, which rises rapidly to a peak as the depth increases, and then drops rapidly. This allows electron radiotherapy to concentrate the dose on the tumor area and reduce damage to deep normal tissues. Electrons, like photons, also indirectly damage cells by producing free radicals, but because electrons have low energy, their damage to cells is mainly concentrated in the shallow areas of the tumor. Electron radiotherapy is mainly used to treat superficial tumors such as skin cancer and early lesions of breast cancer. Electron beams also have a very good effect on postoperative scar hyperplasia.

[0032] Compared with traditional radiotherapy techniques, FALSH can release extremely high radiation doses in a very short time (such as nanoseconds), thereby inducing unique biological responses in tumor cells, such as rapid accumulation of DNA damage and failure of repair mechanisms. This unique radiotherapy method not only improves the killing effect on tumor cells, but also reduces damage to normal tissues.

[0033] The present invention encapsulates garcinia acid (GA) in nanoliposomes to prepare GL, which significantly improves the shortcomings of garcinia acid, such as poor water solubility and poor stability; the prepared GL has good biocompatibility and passive targeting, can efficiently deliver garcinia acid to the tumor site, and reduce the distribution of the drug in normal tissues, thereby improving the anti-tumor effect of the drug and reducing side effects.

[0034] The combined treatment strategy of the present invention is to combine GL with X-FALSH radiotherapy technology to achieve a double attack on tumor cells. The synergistic effect of the two has a significant therapeutic effect in cancer treatment, which not only significantly enhances the anti-tumor effect, but also reduces the side effects during the treatment process and improves the patient's quality of life.

[0035] Since GL has a wide range of applicability and can be loaded with different chemotherapy drugs, and X-FALSH radiotherapy technology is also applicable to a variety of tumor types, the combined treatment strategy of the present invention is expected to become a universal cancer treatment strategy, bringing more treatment options and hope to cancer patients.

[0036] In an optional embodiment of the present invention, the cancer is selected from at least one of pancreatic cancer, breast cancer, melanoma, gastric cancer, liver cancer, cervical cancer, ovarian cancer, lymphoma and brain tumor.

[0037] Pancreatic cancer has an insidious onset and rapid metastasis, and is known as the "king of cancer". Studies have shown (Tu Kerong, Yang Shangqin, Xu Qiqi, et al. Progress in the pharmacological mechanism and toxicology of garcinia acid [JL]. Chinese Journal of Traditional Chinese Medicine, 2024: 1-12. DOI: 10.19540 / j.cnki.cjcmm.20240918.602.) that garcinia acid can target GPR108 and promote its degradation, and then inhibit pancreatic cancer growth by blocking NF-κB signaling. In addition, garcinia acid can increase the formation of acidic vesicle organelles and autophagosomes and increase autophagic flux in pancreatic cancer PANC-1 cells and xenograft tumor tissues, and its molecular mechanism involves inducing the expression of LC3-II and Beclin-1 proteins and reducing the expression of p62 protein. In addition, the effects of garcinia acid on reducing mitochondrial membrane potential and promoting ROS production contribute to the activation of the cellular autophagy process. Clinically, gemcitabine resistance greatly restricts the efficacy of pancreatic cancer chemotherapy. Studies have shown that gambogic acid can reduce the expression levels of ribonucleotide reductase subunit-M2 (RRM2) protein and mRNA by inhibiting the ERK / cell cycle-related transcription factor (E2F1) pathway, and this effect is closely related to reversing gemcitabine resistance.

[0038] Breast cancer, also known as breast cancer, is a common malignant tumor in women, which arises from the uncontrolled proliferation of mammary epithelial cells.

[0039] Melanoma usually refers to malignant melanoma, which is a highly malignant tumor derived from melanocytes, abbreviated as melanoma. It mostly occurs in the skin, but can also be seen in mucous membranes and internal organs, accounting for about 3% of all tumors.

[0040] Gastric cancer is a malignant tumor originating from the gastric mucosal epithelium. There are obvious regional differences in the incidence of gastric cancer. The incidence of gastric cancer in the northwest and eastern coastal areas of my country is significantly higher than that in the southern region.

[0041] Liver cancer is a malignant tumor of the liver, which can be divided into two categories: primary and secondary. Primary malignant liver tumors originate from the epithelial or mesenchymal tissue of the liver. The former is called primary liver cancer, which is a common and extremely harmful malignant tumor; the latter is called sarcoma, which is relatively rare compared with primary liver cancer. Secondary or metastatic liver cancer refers to malignant tumors originating from multiple organs throughout the body that invade the liver.

[0042] Cervical cancer, also known as cervical cancer, is a malignant tumor of the female reproductive tract that occurs in the cervix of the uterus. The most common type is squamous cell carcinoma.

[0043] Ovarian cancer is a type of malignant tumor of the ovary, which refers to a malignant tumor that grows on the ovary. 90%-95% of them are primary ovarian cancers, and the other 5%-10% are primary cancers from other parts of the body that have metastasized to the ovary.

[0044] Brain tumors are tumors that grow inside the skull, including primary brain tumors that arise from the brain parenchyma and secondary brain tumors that metastasize to the skull from other parts of the body.

[0045] Furthermore, in an optional embodiment of the present invention, the technology of combining GL with X-FALSH radiotherapy technology is used to treat pancreatic cancer and breast cancer, and both have achieved remarkable results, which is expected to bring more treatment options and hope to cancer patients and open up new avenues for cancer treatment.

[0046] It should be noted that in other optional embodiments of the present invention, the technology of combining GL with X-FALSH radiotherapy technology can be selected according to actual needs to treat any one of melanoma, gastric cancer, liver cancer, cervical cancer, ovarian cancer and brain tumor.

[0047] In an optional embodiment of the present invention, cancer treatment includes at least one of the following application methods:

[0048] (1) Inhibit tumor growth;

[0049] (2) The subject's body weight remains stable;

[0050] (3) Inhibit inflammatory response;

[0051] (4) Promote the production of anti-tumor and anti-viral cytokines;

[0052] (5) Inhibit the production of transforming growth factor β (TGF-β).

[0053] It should be noted that inhibiting tumor growth includes inhibiting tumor volume growth and mass growth. The calculation of tumor volume is as follows: Tumor volume is measured using a vernier caliper, and the measurement data includes the longest diameter of the transplanted tumor (a, mm) and the widest diameter perpendicular to the longest diameter (b, mm). According to the formula V (mm 3 )=1 / 2×a×b 2 Tumor volume was calculated.

[0054] In an optional embodiment of the present invention, inhibiting the inflammatory response includes inhibiting the production of proinflammatory cytokines, wherein the proinflammatory cytokines include at least one of TNF-α, IL-1β, IL-2, IL-6, IL-8, IL-12, IFN-γ and macrophage migration inhibitory factor (MIF).

[0055] TNF-α is an important pro-inflammatory cytokine that participates in acute inflammatory responses. It can induce tumor cell apoptosis, inhibit angiogenesis, and enhance immune responses. IL-1β is a pro-inflammatory cytokine that can promote the activation and migration of immune cells and plays an important role in the initiation and maintenance of inflammatory responses.

[0056] In an optional embodiment of the present invention, inhibiting inflammatory response also includes promoting the production of anti-inflammatory cytokines, wherein the anti-inflammatory cytokines include at least one of IL-4, IL-13, IL-10 and IL-1 receptor antagonist (IL-1Ra).

[0057] IL-10 is an important anti-inflammatory cytokine that controls inflammatory responses and maintains the balance of the immune system mainly by inhibiting the production of pro-inflammatory cytokines.

[0058] Anti-tumor and anti-viral cytokines include IFN-γ, which is a key anti-tumor and anti-viral cytokine that can activate macrophages and natural killer cells (NK cells) and promote cytotoxic T cells (CTLs) to kill tumor cells.

[0059] Transforming growth factor β (TGF-β), TGF-β1 is a multifunctional cytokine with immunosuppressive effects that can inhibit the activation of T cells and NK cells while promoting the formation of extracellular matrix.

[0060] In an optional embodiment of the present invention, the radiotherapy method is photon-ultra-high dose rate radiotherapy (X-FLASH).

[0061] The irradiation dose and dose rate of photon-ultra-high dose rate radiotherapy depend on the type, location, size of the tumor, and the overall health of the patient. Reasonable adjustments should be made based on actual needs.

[0062] In an optional embodiment of the present invention, the administration method of the nanodrug (GL) is selected from any one of intravenous injection, intramuscular injection, and oral administration.

[0063] It should be noted that in order to allow the drug to better exert its therapeutic effect and reduce adverse reactions, the administration method of GL in the embodiment of the present invention can be selected in different ways according to the overall health condition of the patient.

[0064] Intravenous administration is the process of delivering drugs into the body through veins. It is generally fast and patients can accept it quickly. However, due to the different thicknesses of blood vessels, there will be certain differences in the concentration of the drug.

[0065] Intramuscular injection is the injection of drugs into the body through muscle tissue. Generally, the injection site is shallow, so the effect is quicker. However, due to the large amount of drug injected, the drug is distributed over a wider area in the body, and the therapeutic effect achieved will be better.

[0066] Oral administration is to allow drugs to enter the body through oral administration and be absorbed through the gastrointestinal tract to achieve the effect of treating diseases. However, due to the presence of food in the gastrointestinal tract, the drug is easily decomposed after entering the stomach, making it unable to achieve the effect of treating diseases. Therefore, it is recommended that patients take oral drugs after meals as much as possible to reduce the irritation of the drug to the gastrointestinal tract.

[0067] In a second aspect, an embodiment of the present invention provides a nanomedicine as described above, wherein the nanomedicine comprises gambogic acid, cholesterol and lecithin in a mass ratio of 1:(1.5-2.5):(14-15).

[0068] It should be noted that the proportion of raw materials in GL can ensure the particle size, oxidative stability and physical stability of liposomes. The unique composition of liposomes is not only conducive to controlled release and targeted delivery, but also enhances biocompatibility, making lipid nanoparticles a pioneering force in modern medicine.

[0069] Specifically, the mass ratio of gambogic acid, cholesterol and lecithin is selected from any one of 1:1.5:14, 1:1.5:15, 1:1.8:14.5, 1:1.8:15 or 1:2:15, or other values ​​within the range of 1:(1.5-2.5):(14-15).

[0070] Among them, cholesterol can ensure the stability and fluidity of the membrane. Its addition is very suitable for applications that require extended circulation time and enhanced structural integrity. The molecular geometry of cholesterol derivatives can further affect the delivery efficiency and biodistribution of lipid nanoparticles. Cholesterol can regulate the fluidity of the phospholipid bilayer membrane. If it is added beyond the range and exceeds the membrane load, it will cause reduced membrane permeability, partial liposome rupture, and drug leakage.

[0071] Phosphatidylcholine provides the amphiphilicity necessary for the formation of a lipid bilayer in the nanoparticles, helping to improve the stability and biocompatibility of the formulation.

[0072] In an optional embodiment of the present invention, GA is encapsulated in nanoliposomes to form a stable and uniform GL solution, which can significantly improve the shortcomings of poor water solubility and poor stability of GA, thereby enabling it to better exert its anti-tumor effect. GA-loaded nanoliposomes are an innovative nanomedicine with good biocompatibility and passive targeting, which can efficiently deliver GA to the tumor site and reduce the distribution of the drug in normal tissues, thereby improving the anti-tumor effect of the drug and reducing side effects.

[0073] In an optional embodiment of the present invention, the preparation of nanomedicine includes the following steps: dissolving gambogic acid, cholesterol and lecithin in an alcohol solution, mixing at a temperature of 35°C-50°C to obtain a film; then hydrating the film and filtering to obtain GL.

[0074] In an optional embodiment of the present invention, the film hydration treatment includes ultrasonic treatment in an intermittent circulation manner;

[0075] The interval cycle mode includes ultrasonic treatment for 3s-6s and pause treatment for 1s-3s in sequence, such as ultrasonic treatment for 4s, pause for 1s, ultrasonic treatment for 4s, pause for 1s, ultrasonic treatment for 4s, and pause for 1s in sequence. The total ultrasonic treatment time and the interval cycle time can be reasonably set according to the actual amount of raw materials processed.

[0076] It should be noted that ultrasonic technology generates energy through high-frequency vibration, which will produce strong cavitation when propagating in liquid, causing liquid molecules to move at high speed. This high-speed movement can accelerate the movement of water molecules, thereby improving hydration efficiency, making the film hydrated more thoroughly and facilitating filtration treatment.

[0077] In an optional embodiment of the present invention, the alcohol solution used in the process of preparing the film is selected from any one of methanol, anhydrous ethanol and propanol; further, the alcohol solution is anhydrous ethanol solution. The film preparation process requires heating treatment, mainly to make the raw materials completely dissolved and the reaction more thorough.

[0078] The film hydration treatment is to add water to the prepared film system for ultrasonic treatment, wherein the volume ratio of the alcohol solution to the water in the film hydration treatment is 1:(1-2), and further, the volume ratio is 1:(1.2-1.6).

[0079] The type of water can be selected from pure water, ultrapure water, deionized water, distilled water, etc. according to actual needs; adding too much water will increase the time of subsequent filtration treatment, while adding too little water will not completely hydrate the film and most of the liposomes will be filtered out, resulting in a waste of raw materials.

[0080] In an optional embodiment of the present invention, the prepared liposome solution is subjected to extrusion filtration, specifically, the hydrated liposome initial solution is subjected to extrusion filtration through an extruder and a filter membrane with a pore size of 0.1 μm-0.45 μm to obtain a nanoliposome solution loaded with gambogic acid, i.e., GL. In a preferred embodiment of the present invention, a filter membrane with a pore size of 0.45 μm is used for the first time, and a filter membrane with a pore size of 0.22 μm is used for the second time. Filter membranes with other pore sizes can also be selected for filtration according to actual needs.

[0081] It should be noted that during the liposome preparation process, the main function of the filter membrane is to disperse the original suspension of liposomes to form liposome particles with uniform particle size distribution. The filter membrane screens the particles through its pore structure, allowing only particles smaller than the pore size of the filter membrane to pass through, thereby achieving particle dispersion and screening. Secondly, multiple uses of the filter membrane can effectively filter out impurities generated during the liposome preparation process, such as unreacted raw materials.

[0082] Specifically, the preparation method of GL includes the following preparation steps:

[0083] Weigh GA, cholesterol and lecithin in proportion and place in a rotary evaporator. Then add anhydrous ethanol and perform vacuum rotary evaporation for 3.5h-4.5h at a temperature of 35℃-50℃ and a rotation speed of 45r / min-65r / min to form a uniform film.

[0084] Add water in a corresponding proportion to the membrane system for hydration treatment, and the hydration time is 1.0h-2.0h. Then, ultrasonic treatment is carried out under the condition of 25%-40% power, using a 3s-6s ultrasonic and 1s-3s interval cycle. The obtained solution is filtered through 0.45μm and 0.22μm filter membranes in turn to obtain GL solution.

[0085] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0086] Example 1

[0087] This embodiment provides a nanoliposome loaded with gambogic acid, and the preparation method thereof comprises the following steps:

[0088] The raw materials were weighed in proportion and placed in a 250 mL rotary evaporation bottle, including 5 mg GA, 9.23 mg cholesterol and 72.43 mg lecithin. 5 mL of anhydrous ethanol was added and vacuum rotary evaporation was performed at a temperature of 40°C and a rotation speed of 50 r / min for 4 hours to form a uniform film.

[0089] Then, 7 mL of pure water was added, and the film was hydrated for 1.5 h.

[0090] The hydrated film was ultrasonically treated at a power of 30%, using a cycle of 5 s ultrasonic treatment and 1 s interval, with a total ultrasonic time of 5 min.

[0091] The solution obtained by ultrasound was filtered through 0.45 μm and 0.22 μm filter membranes in turn to obtain a nanoliposome solution loaded with gambogic acid, namely, a GL solution.

[0092] Test Example 1

[0093] In this test example, the GL solution prepared in Example 1 was observed for appearance and tested for microscopic morphology, wherein the microscopic morphology test was to take microscopic structural images of the sample using a transmission electron microscope (TEM) to characterize the morphology and size of the GL, and the test instrument model was JEM-1400FLASH. The specific results were: Figure 1 is the appearance of GL solution, Figure 2 It is the microscopic form of GL.

[0094] from Figure 1 It can be seen that the solution of GL is a yellow clear liquid, which dissolves evenly in the aqueous solution, indicating that the GL prepared in this example can effectively improve the shortcomings of poor water solubility and poor stability of Garcinia acid.

[0095] from Figure 2 It can be seen that GL has a uniform spherical structure and good distribution characteristics, indicating that it has good consistency in morphology and size distribution; the average diameter of GL is 120 nm, indicating that it is suitable as a carrier for drug delivery systems.

[0096] Experimental Example 1

[0097] This experimental example screens the X-FLASH radiation dose, which includes the following implementation process:

[0098] 1.1 Construction of subcutaneous pancreatic cancer transplant model

[0099] Mouse pancreatic cancer Panc-02 cells were cultured in RPMI 1640 medium (containing 10% fetal bovine serum and 1% penicillin-streptomycin) at 37°C in a constant temperature incubator with 5% CO2. Cells were collected by centrifugation and the cell concentration was adjusted to 1×10 7 5-week-old healthy C57BL / 6 mice were selected to establish a Panc-02 cell subcutaneous transplant tumor model. After confirming that the mice were in good condition, the backs of the mice were disinfected with iodine, and 5×10 5 Panc-02 cells were inoculated and the tumor growth was observed every 3 days. When the tumor volume of the mouse grew to about 100 mm 3 Carry out the subsequent experimental plan.

[0100] 1.2X-FLASH radiation dose screening

[0101] After constructing the subcutaneous transplanted tumor mouse model, the mice were randomly divided into the control group (no treatment), 5Gy group, 10Gy group, 15Gy group and 20Gy group according to the tumor volume, with 10 mice in each group. The mice were irradiated at a relatively safe dose rate, and each group of mice was irradiated once on the 4th day. After the experiment, the mice were killed by cervical dislocation, and the tumor was removed, weighed and recorded.

[0102] It should be noted that the safer dose rate is selected from the acceptable dose rate of 200Gy / s-210Gy / s within the safety range; in this embodiment, the dose rate selected based on actual conditions is 207Gy / s. In other optional embodiments of the present invention, other values ​​within the range of 200Gy / s-210Gy / s can be selected according to actual needs.

[0103] 1.3 Experimental Results

[0104] Tumor inhibition rate = [(average tumor mass in the control group - average tumor mass in the treatment group) / average tumor mass in the control group] × 100%, see Figure 3 .

[0105] Figure 3 The results of X-FLASH radiation dose screening are intuitively expressed by drawing a bar graph of mouse tumor inhibition rate. The bar graph is the tumor inhibition rate calculated based on the tumor mass obtained by subsequent dissection after the experiment, when mice were killed by cervical dislocation in accordance with ethical standards.

[0106] The analysis results showed that with the increase of X-FLASH radiation dose, the tumor inhibition rate of mice showed an upward trend. Among them, the tumor inhibition rates of the 15Gy group and the 20Gy group were quite close, both reaching a high inhibition level. However, considering that the increase of X-FLASH dose may have an adverse effect on the overall health of mice, in order to achieve the effect of pursuing the therapeutic effect while minimizing the physiological interference of experimental animals, this test case finally selected 15Gy as the appropriate treatment dose of X-FLASH.

[0107] Experimental Example 2

[0108] In this experimental example, GL prepared in Example 1 is combined with X-FLASH to treat pancreatic cancer. The specific implementation process is as follows:

[0109] 2.1 Construction of subcutaneous pancreatic cancer transplant model

[0110] The implementation process is the same as that of Experimental Example 1.

[0111] 2.2 Design of experimental conditions for combined treatment of pancreatic cancer with GL and X-FLASH

[0112] After constructing the subcutaneous transplanted tumor mouse model, the mice were randomly divided into the Control group, GL group, X-FLASH group, X-FLASH+GL group, and CX-FLASH (conventional irradiation) group according to the tumor volume, with 10 mice in each group. The design of each group is as follows:

[0113] Control group: only 100 μL of normal saline was injected into the tail vein every 2 days for 6 consecutive times.

[0114] GL group: 100 μL of 2 mg / kg GL was injected into the tail vein every 2 days for 6 consecutive times.

[0115] X-FLASH group: X-FLASH was given only once on the 4th day, with a dose of 15 Gy and a dose rate of 207 Gy / s.

[0116] X-FLASH+GL group: 100 μL of 2 mg / kg GL was injected into the tail vein every 2 days for 6 consecutive times, and X-FLASH was irradiated once on the 4th day with a dose of 15 Gy and a dose rate of 207 Gy / s.

[0117] CX-FLASH group: conventional X-FLASH was given only once on the 4th day, with a dose of 15 Gy and a dose rate of 207 Gy / s.

[0118] It should be noted that the difference between the X-FLASH+GL group and the CX-FLASH group is in the dose rate. X-FLASH can deliver an irradiation dose higher than 8Gy, with a dose rate exceeding 40Gy / s, while the dose rate of conventional irradiation is only about 1-7Gy / s. In terms of irradiation time, X-FLASH can provide an ultra-high radiation dose to the irradiated target area in a very short time (less than 1s), while the conventional irradiation time takes several minutes. In other experimental examples of the present invention, the instrument can be reasonably adjusted according to actual needs, and the corresponding irradiation dose and dose rate also need to be adjusted.

[0119] During the administration period, the weight and tumor volume of mice were recorded every 3 days. The tumor volume was measured with a vernier caliper, and the measurement data included the longest diameter of the transplanted tumor (a, mm) and the widest diameter perpendicular to the longest diameter (b, mm). According to the formula V (mm 3 )=1 / 2×a×b 2 Tumor volume was calculated.

[0120] 2.3 Evaluation of pharmacodynamics after treatment

[0121] 2.3.1 Changes in tumor volume and body weight of mice during combined treatment of pancreatic cancer

[0122] The results of the tumor volume growth curve of mice during combined treatment of pancreatic cancer are shown in Figure 4 (a) Figure 4 The results of (a) show that during the combined treatment of pancreatic cancer, the tumor volume of the treatment group was smaller than that of the Control group. This result shows that different treatment groups inhibited the growth of mouse tumors. Compared with other groups, the tumor growth rate of the GL+X-FLASH group has always been at the lowest level, lower than other groups. This result shows that the GL+X-FLASH group has the best anti-tumor effect, and GL and X-FLASH have a synergistic effect.

[0123] The results of the weight change curve of mice during combined treatment of pancreatic cancer are shown in Figure 4 (b) Figure 4 (b) The overall health status of mice was assessed by their body weight during the combined treatment of pancreatic cancer. The body weight of mice in different treatment groups remained basically stable during the experiment, indicating that each treatment regimen had no significant effect on the physiological state of mice.

[0124] 2.3.2 Tumor morphology and volume of mice after combined treatment of pancreatic cancer

[0125] Results related to tumor morphology in mice after combined treatment of pancreatic cancer are shown in Figure 5 (a) Figure 5(a) After the combined treatment of pancreatic cancer, the mice were killed according to ethical requirements, and the tumor photos were dissected. The tumor volume of the treatment group was smaller than that of the control group. This result shows that the different treatment groups significantly inhibited the growth of mouse tumors. Compared with other groups, the tumor volume of the GL+X-FLASH group was the smallest, smaller than other groups. This result shows that the GL+X-FLASH group has the best anti-tumor effect, and GL and X-FLASH have a synergistic effect.

[0126] The results related to the tumor volume of mice after the combined treatment are shown in Figure 5 (b) Figure 5 (b) After the combined treatment of pancreatic cancer, the mice were killed according to ethical requirements, and the tumor volume was dissected. As can be seen from the figure, the tumor volume of the treatment group was smaller than that of the control group. This result shows that different treatment groups inhibited the growth of mouse tumors. Compared with other groups, the tumor volume of the GL+X-FLASH group was the smallest, with an average tumor volume of 95.76±54.07mm 3 , which is much smaller than that of other groups. This result shows that the anti-tumor effect of GL+X-FLASH group is the best, and the liposomes of gambogic acid and X-FLASH have a synergistic effect.

[0127] 2.3.2 Tumor mass and tumor inhibition in mice after combined treatment of pancreatic cancer

[0128] After the combined treatment of pancreatic cancer, the mice were killed by cervical dislocation, the tumors were removed, weighed and recorded, the mouse tumor volume growth curve and the mouse weight change curve were drawn, the tumor mass was weighed and the tumor inhibition rate was calculated. Tumor inhibition rate = [(Control group average tumor mass - treatment group average tumor mass) / Control group average tumor mass] × 100%.

[0129] Results related to mouse weight after combined treatment are shown in Figure 6 (a) Figure 6 (a) is the tumor mass obtained after the mice were killed according to ethical requirements after the combined treatment of pancreatic cancer. As can be seen from the figure, the tumor mass of the treatment group was smaller than that of the Control group. This result shows that different treatment groups inhibited the growth of mouse tumors. Compared with other groups, the tumor mass of the GL+X-FLASH group was the smallest, with an average tumor mass of 0.1594±0.0425g, which was smaller than that of other groups. This result shows that the GL+X-FLASH group has the best anti-tumor effect, and the gambogic acid liposomes and X-FLASH have a synergistic effect.

[0130] The results of tumor inhibition rate in mice after combined treatment are shown in Figure 6 (b) Figure 6(b) After the combined treatment of pancreatic cancer, the mice were killed according to ethical requirements, and the tumor inhibition rate was calculated based on the tumor mass dissected out. It showed that different treatment groups significantly inhibited the growth of mouse tumors. Among them, the GL+X-FLASH group showed the highest tumor inhibition rate, indicating that the combined treatment has a synergistic effect and has the strongest inhibitory effect on mouse tumors.

[0131] 2.3.3 Detection of physiological indicators after combined treatment of pancreatic cancer and evaluation of biosafety in mice

[0132] 2.3.3.1 Experimental Materials

[0133] IL-6: Nanjing Jiancheng Biotechnology Co., Ltd., catalog number H007-1-2;

[0134] TNF-α: Nanjing Jiancheng Biotechnology Co., Ltd., catalog number H052-1-2;

[0135] IL-1β: Nanjing Jiancheng Biotechnology Co., Ltd., catalog number H002-1-2;

[0136] IFN-γ: Nanjing Jiancheng Biotechnology Co., Ltd., catalog number H025-1-2;

[0137] IL-10: Nanjing Jiancheng Biotechnology Co., Ltd., catalog number H009-1-2;

[0138] TGF-β1: Nanjing Jiancheng Biotechnology Co., Ltd., catalog number H034-1-1.

[0139] 2.3.4.2 Detection of mouse physiological indicators and assessment of mouse biosafety

[0140] (1) ELISA immunoassay and results

[0141] After the combined treatment of pancreatic cancer, the blood of mice was collected, the samples were allowed to stand for 30 min, and then centrifuged at 1000 g for 15 min. After separating red blood cells (RBCs), the supernatant containing serum was collected and ELISA was performed to measure physiological indicators such as TNF-α, IL-1β, IFN-γ, IL-10 and TGF-β1.

[0142] The serum TNF-α level of mice after combined treatment of pancreatic cancer was Figure 7 (a), TNF-α is an important pro-inflammatory cytokine involved in acute inflammatory response, which can induce tumor cell apoptosis, inhibit angiogenesis and enhance immune response. The figure shows that different treatment groups significantly reduced the TNF-α concentration in mouse serum, among which the GL and X-FLASH combined treatment group had the lowest TNF-α level, indicating that these treatments are effective in inhibiting inflammatory response, among which the combined treatment group has the strongest anti-inflammatory effect.

[0143] The serum IL-1β level of mice after combined treatment of pancreatic cancer was Figure 7 (b), IL-1β is a pro-inflammatory cytokine that can promote the activation and migration of immune cells and plays an important role in the initiation and maintenance of inflammatory responses. The figure shows that different treatment groups significantly reduced the IL-1β concentration in mouse serum, among which the GL and X-FLASH combined treatment group had the lowest IL-1β level, indicating that these treatments are effective in inhibiting inflammatory responses, and the combined treatment group has the strongest anti-inflammatory effect.

[0144] The serum IFN-γ level of mice after combined treatment of pancreatic cancer was Figure 7 (c), IFN-γ is a key anti-tumor and anti-viral cytokine that can activate macrophages and natural killer cells (NK cells) and promote cytotoxic T cells (CTLs) to kill tumor cells. From the Control group to the CX-FLASH group, the concentration of IFN-γ gradually increased, indicating that the induction effect of different treatments on IFN-γ gradually increased, especially X-FLASH+GL and CX-FLASH treatments showed stronger immune activation effects.

[0145] The serum IL-10 level of mice after combined treatment of pancreatic cancer Figure 7 (d), IL-10 is an important anti-inflammatory cytokine that controls inflammatory responses and maintains the balance of the immune system mainly by inhibiting the production of pro-inflammatory cytokines. The figure shows that different treatment groups significantly increased the IL-10 concentration in mouse serum, among which the GL and X-FLASH combined treatment group had the highest IL-10 level, indicating that these treatments are effective in controlling inflammatory responses, and the combined treatment group has the strongest anti-inflammatory effect.

[0146] The serum TGF-β1 level of mice after combined treatment of pancreatic cancer Figure 7 (e), TGF-β1 is a multifunctional cytokine with immunosuppressive effects. It can inhibit the activation of T cells and NK cells and promote the formation of extracellular matrix. The figure shows that different treatment groups significantly reduced the concentration of TGF-β1 in mouse serum, among which the TGF-β1 level in the GL and X-FLASH combined treatment group was the lowest, indicating that these treatments all have immunosuppressive effects, among which the combined treatment group has the strongest immunosuppressive effect.

[0147] (2) Blood routine and blood biochemical tests and corresponding results

[0148] Routine blood test instruments: LSC-316C vertical refrigerated display cabinet (Xingxing), BC-5000vet five-category blood cell analyzer (Mindray).

[0149] Blood biochemical test instrument: Chemray 240 fully automatic biochemical analyzer (Shenzhen Raydu Life Science).

[0150] The blood routine results of mice after combined treatment of pancreatic cancer were shown in Figure 8 , from left to right: (a) percentage of white blood cells, neutrophils, lymphocytes, monocytes; (b) hemoglobin, mean corpuscular hemoglobin concentration; (c) red blood cells, platelets; compared with the control group mice, the treated group mice had Figure 8 The indicators in (a)-8(c) were all within the normal range, and there was no statistical difference between the groups. This indicates that the GL combined with X-FLASH treatment strategy does not cause adverse reactions such as inflammation and bleeding.

[0151] The results of blood biochemistry of mice after combined treatment of pancreatic cancer were shown in Fig. 9 Compared with the control group, the liver and kidney functions of the treated group mice were significantly improved. Fig. 9 (a)-9(g) indicators are all within the normal range, and Fig. 9 The Urea index in (h) increased after GL administration, but X-FLASH had no effect on it. This indicates that GL and X-FLASH do not cause damage to the liver, but may cause damage to the kidneys due to GL's metabolism in the kidneys.

[0152] (3) H&E staining and results

[0153] After the combined treatment of pancreatic cancer, the tumor tissue was fixed, placed in an embedding box, dehydrated with ethanol, transparentized with xylene, embedded in wax, sliced ​​and mounted (thickness of about 5 μm), and dried. Dewaxing and hydration, hematoxylin staining for 5 minutes, eosin staining for 3 minutes, ethanol dehydration, xylene transparent, and neutral resin sealing. Test instruments: TSJ-SD dehydrator (Changzhou Zhongwei Electronic Instrument Co., Ltd.), BMJ-A embedding machine (Changzhou Zhongwei Electronic Instrument Co., Ltd.), SHANDON FINESSE 325 pathology slicer (Thermo Fisher Scientific Co., Ltd.), JB-L5 freezing table (Wuhan Junjie Electronic Co., Ltd.), PHY-Ⅲ tissue spreader (Changzhou Zhongwei Electronic Instrument Co., Ltd.), BP0510 anti-slip slide (Hubei Bios Biotechnology Co., Ltd.), CX-31 upright microscope (Olympus Co., Ltd.), (Hamamatsu Photonics Co., Ltd., Japan).

[0154] H&E staining of mouse tumor tissue after combined treatment of pancreatic cancer. Fig.10The results of H&E staining of important organs such as the heart, liver, spleen, lung, and kidney in the Control group and different treatment groups showed that the cardiomyocytes of all mice were neatly arranged, the nuclei, endocardium, and exomembranes were clearly stained, and the cytoplasm was evenly stained. The hepatocytes and spleen cells were closely arranged, the nuclei and vascular systems were clearly stained, and the cytoplasm was evenly stained. In the lung tissue, there were no obvious abnormalities in the alveoli and alveolar walls, and the vascular system was clearly stained. In the kidney tissue, there were no obvious abnormalities in the renal corpuscles and renal tubules, and the renal vascular system was clearly stained. In short, the morphology of the important organs of mice after GL or X-FLASH treatment was not significantly different from that of the Control group, with clear tissue structure, normal cell morphology, and no obvious inflammation, necrosis, or other abnormal changes, indicating that there was no chronic pathological toxicity and side effects of GL and X-FLASH treatment.

[0155] Experimental Example 3

[0156] In this experimental example, GL prepared in Example 1 is combined with X-FLASH to treat breast cancer. The specific implementation process is as follows:

[0157] 3.1 Construction of subcutaneous breast cancer transplant model

[0158] Murine breast cancer 4T1 cells were cultured in RPMI 1640 medium (containing 10% fetal bovine serum and 1% penicillin-streptomycin) at 37°C in a constant temperature incubator with 5% CO. Cells were collected by centrifugation and the cell concentration was adjusted to 1×10 7 5-week-old healthy C57BL / 6 mice were selected to establish a 4T1 cell subcutaneous transplant tumor model. After confirming that the mice were in good condition, the back of the mice was disinfected with iodine, and 5×10 5 4T1 cells were inoculated and the tumor growth was observed every 3 days. When the tumor volume of the mouse grew to about 100 mm 3 Carry out subsequent experimental plan processing.

[0159] 3.2X-FLASH radiation dose screening

[0160] It should be noted that the radiation dose of X-FLASH in the combined treatment of breast cancer with GL and X-FLASH is the same as the screening process of the radiation dose for the treatment of pancreatic cancer in 1.2 of Experimental Example 1.

[0161] 3.3 Design of experimental conditions for combined treatment of breast cancer with GL and X-FLASH

[0162] The design of the experimental conditions for the combined treatment of pancreatic cancer with 2.2GL and X-FLASH is the same as that in Experimental Example 2.

[0163] 3.4 Experimental Results

[0164] 3.4.1 Changes in tumor volume and body weight of mice during combined treatment of breast cancer

[0165] The growth curve of mouse tumor volume after combined treatment of breast cancer is shown in Fig.11 (a) This figure shows that during the treatment, the tumor volume of the treatment group was smaller than that of the Control group. This result indicates that different treatment groups inhibited the growth of mouse tumors.

[0166] The weight change curve of mice during combined treatment of breast cancer is shown in Fig.11 (b) This figure evaluates the overall health status of mice based on their body weight during the treatment process. The body weight of mice in different treatment groups remained basically stable during the experiment, indicating that each treatment regimen had no significant effect on the physiological state of mice.

[0167] 3.4.2 Tumor volume and tumor mass of mice after combined treatment of breast cancer

[0168] After combined treatment of breast cancer, the tumor volume of mice Fig.12 (a) This figure shows the tumor volume obtained after the mice were killed according to ethical requirements after the treatment. It can be seen from the figure that the tumor volume of the treatment group is smaller than that of the control group. This result shows that different treatment groups inhibit the growth of mouse tumors.

[0169] Tumor weight of mice after combined treatment of breast cancer Fig.12 (b) This figure shows the tumor weight obtained after the mice were killed according to ethical requirements after the treatment. It can be seen from the figure that the tumor weights of the different treatment groups are all smaller than those of the Control group. This result indicates that the different treatment groups all inhibit the growth of mouse tumors.

[0170] 3.4.3 Detection of physiological indicators of mice after combined treatment of breast cancer

[0171] 3.4.3.1 ELISA immunoassay

[0172] After the combined treatment of breast cancer, the blood of mice was collected, the samples were allowed to stand for 30 min, and then centrifuged at 1000 g for 15 min. After separating red blood cells (RBCs), the supernatant containing serum was collected and ELISA was performed to measure physiological indicators such as TNF-α, IL-1β, IFN-γ, IL-10 and TGF-β1.

[0173] 3.4.3.2 ELISA immunoassay results

[0174] After combined treatment of breast cancer, the serum TNF-α level of mice was Fig.13 (a) This figure shows that different treatment groups significantly reduced the TNF-α concentration in mouse serum, indicating that these treatments were effective in inhibiting inflammatory responses.

[0175] After combined treatment of breast cancer, the serum IL-1β level of mice was Fig.13 (b) This figure shows that the different treatment groups significantly reduced the IL-1β concentration in mouse serum, indicating that these treatments are effective in inhibiting inflammatory responses.

[0176] After combined treatment of breast cancer, the level of IFN-γ in the serum of mice was Fig.13 (c) As can be seen from the figure, the IFN-γ levels in the different treatment groups were higher than that in the Control group, indicating that the induction effect of different treatments on IFN-γ gradually increased.

[0177] After combined treatment of breast cancer, the serum IL-10 level of mice was Fig.13 (d) This figure shows that different treatment groups significantly increased the IL-10 concentration in mouse serum, among which the GL and X-FLASH combined treatment group had the highest IL-10 level, indicating that these treatments are effective in controlling inflammatory responses, among which the combined treatment group has the strongest anti-inflammatory effect.

[0178] The serum TGF-β1 level of mice after combined treatment of breast cancer was Fig.13 (e) This figure shows that different treatment groups significantly reduced the concentration of TGF-β1 in mouse serum, among which the combined treatment group with GL and X-FLASH had the lowest level of TGF-β1, indicating that these treatments all had immunosuppressive effects, among which the combined treatment group had the strongest immunosuppressive effect.

[0179] In summary, the nanomedicine (GL) provided by the embodiment of the present invention has good biocompatibility and passive targeting, can efficiently deliver gamboge acid to the tumor site, reduce the distribution of the drug in normal tissues, thereby improving the anti-tumor effect of the drug and reducing side effects. Through the synergistic effect of GL and X-FLASH radiotherapy, a double blow to tumor cells is achieved, which not only significantly enhances the anti-tumor effect, but also reduces the side effects during the treatment process and improves the quality of life of patients. The combined treatment strategy of the present invention effectively overcomes the problem that cancer is insensitive to existing radiotherapy and chemotherapy, achieves a more significant and efficient anti-tumor effect, brings more treatment options and hope to cancer patients, and opens up new ways for cancer treatment.

[0180] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An application of nanomedicine combined with radiotherapy in cancer treatment, characterized in that: The nano drug (GL) is a nano liposome loaded with gambogic acid, and the radiotherapy method is selected from any one of photon-ultra-high dose rate radiotherapy (X-FLASH) and electron-ultra-high dose rate radiotherapy (E-FLASH).

2. The use according to claim 1, characterized in that: The cancer is selected from at least one of pancreatic cancer, breast cancer, melanoma, gastric cancer, liver cancer, cervical cancer, ovarian cancer and brain tumor.

3. The use according to claim 1, characterized in that: Cancer treatment includes at least one of the following: (1) Inhibit tumor growth; (2) The subject's body weight remains stable; (3) Inhibit inflammatory response; (4) Promote the production of anti-tumor and anti-viral cytokines; (5) Inhibit the production of transforming growth factor β (TGF-β).

4. The use according to claim 3, characterized in that: The inhibition of inflammatory response includes inhibiting the generation of proinflammatory cytokines, wherein the proinflammatory cytokines include at least one of TNF-α, IL-1β, IL-2, IL-6, IL-8, IL-12, IFN-γ and macrophage migration inhibitory factor (MIF).

5. The use according to claim 3, characterized in that: The inhibition of inflammatory response also includes promoting the production of anti-inflammatory cytokines, wherein the anti-inflammatory cytokines include at least one of IL-4, IL-13, IL-10 and IL-1 receptor antagonist (IL-1Ra).

6. The use according to claim 1, characterized in that: The radiotherapy method is photon-ultra-high dose rate radiotherapy (X-FLASH).

7. The use according to claim 1, characterized in that: The administration method of the nano drug (GL) is selected from any one of intravenous injection, intramuscular injection and oral administration.

8. A nano drug according to any one of claims 1 to 7, characterized in that: The nano drug comprises gambogic acid, cholesterol and lecithin in a mass ratio of 1:(1.5-2.5):(14-15).

9. The nano drug according to claim 8, characterized in that: The preparation of the nano drug comprises the following steps: dissolving gambogic acid, cholesterol and lecithin in an alcohol solution, mixing at a temperature of 35°C-50°C to obtain a film; then hydrating the film, filtering and obtaining the nano drug (GL); The volume ratio of the alcohol solution to water in the film hydration treatment is 1:(1-2).

10. The nano drug according to claim 9, characterized in that The film hydration treatment comprises ultrasonic treatment in an intermittent circulation manner; The interval cycle includes sequentially cycling ultrasonic treatment for 3s-6s and rest treatment for 1s-3s.

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