Pharmaceutical composition for non-small cell lung cancer radiotherapy sensitization and preparation method thereof

Abstract

The invention discloses a pharmaceutical composition for non-small cell lung cancer radiotherapy sensitization and a preparation method thereof, and relates to the technical field of biological medicine, the pharmaceutical composition is composed of lecithin, cholesterol, docetaxel, plumbagin, a DSPE-mPEG2000 long-acting fragment and a DSPE-PEG2000-FA targeting fragment. The novel double-drug combination strategy can enhance the radiotherapy sensitivity of the non-small cell lung cancer, obtains the synergistic and attenuated synergistic anti-lung cancer effect, realizes radiotherapy sensitization, and improves the radiotherapy and chemotherapy curative effects.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly relates to a pharmaceutical composition for radiosensitization of non-small cell lung cancer and a preparation method thereof. Background Art

[0002] The incidence rate of lung cancer is 12.4%, and the mortality rate is 18.7%, ranking first globally. Among them, non-small cell lung cancer (NSCLC) is the most common pathological type, accounting for about 85% of the pathological types of lung cancer. Since most patients are in the advanced stage at the time of diagnosis, the 5-year survival rate of NSCLC patients only remains between 10% and 20%. Although early screening, early diagnosis, and early treatment have improved the overall prognosis of lung cancer to a certain extent, even the 5-year survival rate of early-stage NSCLC patients who can be surgically resected is only about 60%. Currently, the treatment methods for NSCLC mainly include surgical treatment, radiotherapy, chemotherapy, molecular targeted therapy, immunotherapy, etc. Although the development of molecular targeted therapy and immunotherapy has provided new progress for the clinical treatment of NSCLC and increased the survival rate of NSCLC patients, radiotherapy and chemotherapy are still the main treatment means for NSCLC.

[0003] Radiotherapy uses the ionizing radiation of high-energy rays (X-rays, γ-rays, and charged particles) to directly act on cell DNA to cause cell damage, or indirectly react with water molecules to generate ROS to damage DNA, RNA, proteins, and lipids, inducing cell death. 80% of DNA damage is caused by indirect reactions. Radiotherapy may be required at all stages of lung cancer treatment, and about 60% of NSCLC patients receive radiotherapy or combined radiotherapy and chemotherapy at least once during the course of the disease. Although radiotherapy is widely used in NSCLC and even all cancers due to its special advantage of not being limited by tissue depth, there are also many inherent defects that need to be solved to improve its efficacy. During the treatment process, there are problems such as radiotherapy resistance, damage to surrounding normal tissues by radiotherapy, large side effects of combined radiotherapy and chemotherapy, resistance to targeted therapy, and low overall effective rate of immunotherapy. Therefore, there is an urgent need to find a safer and more effective method to improve the sensitivity of lung cancer to radiotherapy.

[0004] Studies have found that the survival and maintenance of normal cell functions depend on the redox system. Due to their high metabolism and mitochondrial dysfunction, tumor cells have elevated ROS levels, resulting in a redox state different from that of normal cells. To adapt to high levels of ROS, tumor cells also have a powerful antioxidant defense mechanism, so they may evade ROS-induced cell death and reduce the efficacy of radiotherapy. If the treatment effect is improved by increasing the dose of radiotherapy, normal tissues will not be able to tolerate the side effects of the treatment. At the same time, excessive ROS can kill tumor cells by oxidatively damaging biomolecules and activating various cell death pathways. Disrupting intracellular redox homeostasis by promoting the production of ROS or reducing the consumption of ROS can effectively increase oxidative stress and improve the efficacy of radiotherapy. Therefore, enhancing the efficacy of radiotherapy by promoting ROS generation is a promising treatment strategy.

[0005] Docetaxel (DTX), also known as docetaxel, is widely used in all lines of chemotherapy and concurrent chemoradiotherapy for NSCLC. Due to its poor solubility, ethanol or polysorbate 80 is commonly used for solubilization. This preparation can cause short-term and long-term side effects including allergy, febrile neutropenia, fatigue, fluid retention, and peripheral neuropathy. In addition, DTX, as a substrate of the drug efflux pump P-glycoprotein (P-gp), leads to tumor drug resistance and poor bioavailability. There is an urgent need to develop an adjuvant means to increase the therapeutic effect of DTX.

[0006] In recent years, traditional Chinese medicine components have received extensive attention. Plumbagin (PLB) has the effect of killing tumors by promoting the production of ROS and is expected to become a potential candidate drug for radiosensitization and chemosensitization. Moreover, a large number of literature reports have shown that the combination of traditional Chinese medicine with radiotherapy, chemotherapy, or targeted therapy can improve the treatment effect and reduce the occurrence of related adverse reactions and complications. However, most active anti-tumor components of traditional Chinese medicine often have poor water solubility and low bioavailability, which limits their clinical application. To overcome or alleviate these problems, developing a safe and effective nanoformulation that can simultaneously deliver two drugs is the first choice. Summary of the Invention

[0007] In view of the above problems, the present invention provides a pharmaceutical composition for radiosensitization of non-small cell lung cancer and its preparation method. The pharmaceutical composition is a long-circulating targeted co-delivery liposome nanoparticle Lip-PEG-FA@DTX / PLB co-loaded with docetaxel and plumbagin based on the regulation of oxidative stress. This liposome can simultaneously encapsulate docetaxel and plumbagin and is double-modified with polyethylene glycol (PEG) and folic acid (FA). PEGylation can significantly prolong the in vivo circulation time of the drug. At the same time, FA can specifically target the FA receptor on the surface of non-small cell lung cancer, increasing the targeting and drug uptake of the liposome, thereby enhancing the oxidative stress level in lung cancer cells and ultimately achieving the therapeutic radiosensitization and chemosensitization effects on lung cancer.

[0008] To achieve the above object, the technical solution adopted by the present invention is as follows: The present invention provides a pharmaceutical composition for radiosensitization of non-small cell lung cancer, and the pharmaceutical composition is composed of lecithin, cholesterol, docetaxel, plumbagin, DSPE-mPEG 2000 long-acting fragment and DSPE-PEG 2000 -FA targeting fragment.

[0009] The DSPE-PEG 2000 -FA targeting fragment in the liposome acts as a targeting ligand, which can specifically recognize and bind to the folate receptor on the surface of non-small cell lung cancer, increasing the uptake of the liposome; the DSPE-mPEG 2000 long-acting fragment reduces the capture interference of the liposome with albumin and the reticuloendothelial system in the body, and prolongs the circulation time of the liposome in the body.

[0010] Further, the lecithin is egg yolk lecithin.

[0011] Further, the molar ratio of lecithin to cholesterol is 2-10:1, the molar ratio of docetaxel to lecithin is 1:10-50, the molar ratio of docetaxel to plumbagin is 1:10-10:1, the DSPE-mPEG 2000 long-acting fragment and lecithin molar ratio is 1:100-500, the DSPE-PEG 2000 -FA targeting fragment and lecithin molar ratio is 0.2:100-2.

[0012] The present invention also provides a preparation method of a pharmaceutical composition for radiosensitization of non-small cell lung cancer, which specifically includes the following steps: S1: Weigh lecithin, cholesterol, docetaxel, plumbagin and DSPE-mPEG 2000 long-acting fragment and dissolve them in chloroform, mix them at normal pressure for 3-5 min, and after fully mixing the drug and lecithin evenly, slowly evacuate to form a semi-transparent film; S2: Add physiological saline solution to the semi-transparent film, hydrate the film by rotation at 37°C for 2 h to obtain a mixed solution; centrifuge the mixed solution for 10 min, take the supernatant, and then add the DSPE-PEG 2000 -FA targeting fragment to the supernatant and incubate at 37°C for 2 h; S3: After the incubation, the obtained solution is cooled for 8 s by probe sonication for 3 s in an ice-water bath. The cyclic operation is carried out for a total of 15 min, and then sonicated in a water bath at room temperature for 10 min. It is extruded through a liposome extruder through the membrane repeatedly for 10 times to obtain the long-circulating targeted co-delivery liposome nanoparticles Lip-PEG-FA@DTX / PLB co-loaded with docetaxel and plumbagin, that is, the pharmaceutical composition.

[0013] Further, in the step S1, the molar ratio of lecithin to cholesterol is 2-10:1, the molar ratio of docetaxel to lecithin is 1:10-50, the molar ratio of docetaxel to plumbagin is 1:10-10:1, and the DSPE- mPEG 2000 long-acting fragment and lecithin molar ratio is 1:100-500.

[0014] Further, in the step S2, DSPE-PEG 2000 -FA targeting fragment and the lecithin in step S1 molar ratio is 0.2:100-2.

[0015] Further, in the step S2, the centrifugation speed is 5000 rpm.

[0016] Among the nanocarriers for drug delivery, liposomes are non-toxic, non-immunogenic, have a high loading capacity for drugs with different physical and chemical properties, and can be surface-modified to extend the drug half-life and achieve targeted drug delivery, achieving a safe and effective therapeutic effect. Through targeted modification design of liposomes, they have more excellent cell selectivity, cell uptake or tumor penetration ability, and longer blood circulation time, showing great advantages in nanomedicine. Folic acid (FA) liposomes are a type of receptor-mediated active-targeted liposomes. Utilizing the difference in the expression of folate receptors on lung cancer cells (high expression) and normal cells (low expression), as well as the high specificity and high affinity of the binding of folate receptors to folic acid, liposomes can be very efficiently carried into tumor cells. Polyethylene glycol (PEG) is a hydrophilic polymer approved by the US Food and Drug Administration, which can increase the hydrophilicity and steric hindrance of nanoparticles, protect nanoparticles from the influence of the reticuloendothelial system, and prolong the circulation time of nanoparticles by preventing the interaction between nanoparticles and plasma proteins. Therefore, by modifying PEG and FA on nanoparticles and utilizing the ligand-receptor affinity, the NSCLC tumor targeting and blood circulation time of nanoparticles can be improved, realizing precise targeted delivery. The long-circulating targeted co-delivery liposome nanocarrier has a high loading capacity and can accurately deliver drugs to enhance the radiosensitivity.

[0017] Compared with the prior art, the beneficial effects of the present invention are: In the present invention, by screening out plumbagin, a drug that has a synergistic effect with docetaxel, a new clinical idea for the treatment of non-small cell lung cancer is provided. With the assistance of nanotechnology, a long-circulating targeted co-delivery liposome nanoparticle Lip-PEG-FA@DTX / PLB based on regulating oxidative stress is successfully prepared, realizing the co-delivery of docetaxel and plumbagin. The preparation process is simple, and both drugs have a high encapsulation rate, with good physicochemical properties and targeting, demonstrating that the liposome nanoparticle has good anti-tumor efficacy, achieving the effect of "1 + 1 > 2". Based on this, the novel dual-drug combination strategy of the present invention can enhance the radiosensitivity of non-small cell lung cancer, obtain a synergistic anti-lung cancer effect of enhancing efficacy and reducing toxicity, achieve radiotherapy sensitization, and improve the efficacy of radiotherapy and chemotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 is the electron micrograph of the Lip-PEG-FA@DTX / PLB nanoparticles of the present invention; Figure 2 is the dispersion stability of the Lip-PEG-FA@DTX / PLB nanoparticles of the present invention; Figure 3 is the in vitro drug release curves of (A) DTX and (B) PLB of the present invention under different pH conditions; Figure 4 is the cytotoxicity and IC50 value of the Lip-PEG-FA@DTX / PLB nanoparticles of the present invention against A549 cells; Figure 4 A shows the cell survival rate of A549 cells treated with different drugs at the same drug concentration (10 μmol / L), Figure 4 B shows the cell survival rate of A549 cells treated with different drugs at different drug concentrations (1, 2, 4, 8, 12, 15, 20, 25, 30, 40, 50 μmol / L), Figure 4 C shows the IC 50 value, Figure 4 D shows the cell survival rate of A549 cells after different treatment methods (treatment with different drugs and combined radiotherapy). DETAILED DESCRIPTION OF THE EMBODIMENTS

[0019] In order to make the objectives and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below with reference to the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0020] In the following examples, the instruments, reagents, materials, etc. involved, unless otherwise specified, are all conventional instruments, reagents, materials, etc. existing in the prior art and can be obtained through regular commercial channels. The experimental methods, detection methods, etc. involved in the following examples, unless otherwise specified, are all conventional experimental methods, detection methods, etc. existing in the prior art.

[0021] Example 1 This example provides a pharmaceutical composition for radiosensitization of non-small cell lung cancer. The pharmaceutical composition consists of lecithin (egg yolk lecithin), cholesterol, docetaxel, plumbagin, DSPE-mPEG 2000 long-acting fragment and DSPE-PEG 2000 -FA targeting fragment.

[0022] The molar ratio of the above lecithin to cholesterol is 2:1, the molar ratio of docetaxel to lecithin is 1:10, the molar ratio of docetaxel to plumbagin is 1:10, and the molar ratio of DSPE-mPEG 2000 long-acting fragment to lecithin is 1:100, and the molar ratio of DSPE-PEG 2000 -FA targeting fragment to lecithin is 0.2:100.

[0023] The preparation method of the pharmaceutical composition specifically includes the following steps: S1: Weigh lecithin, cholesterol, docetaxel, plumbagin and DSPE-mPEG 2000 long-acting fragment and dissolve them in chloroform. Mix them at normal pressure for 3 min. After the drugs are fully mixed with lecithin, slowly evacuate to form a translucent film. S2: Add physiological saline solution to the translucent film. Hydrate the film by rotation at 37 °C for 2 h to obtain a mixed solution; centrifuge the mixed solution at 5000 rpm for 10 min, take the supernatant, and then add DSPE-PEG 2000 -FA targeting fragment to the supernatant and incubate at 37 °C for 2 h. S3: After the incubation, subject the obtained solution to probe sonication in an ice-water bath for 3 s and cooling for 8 s, and perform the cyclic operation for a total of 15 min. Then perform water bath sonication at room temperature for 10 min, and extrude the solution through a liposome extruder through the membrane repeatedly for 10 times to obtain long-circulating targeted co-delivery liposome nanoparticles Lip-PEG-FA@DTX / PLB co-loading docetaxel and plumbagin, that is, the pharmaceutical composition.

[0024] Example 2 This example provides a pharmaceutical composition for radiosensitization of non-small cell lung cancer. The pharmaceutical composition consists of lecithin (egg yolk lecithin), cholesterol, docetaxel, plumbagin, DSPE-mPEG 2000 long-acting fragment and DSPE-PEG 2000-FA targeting fragment composition.

[0025] The molar ratio of the above-mentioned lecithin to cholesterol is 6:1, the molar ratio of docetaxel to lecithin is 1:30, the molar ratio of docetaxel to plumbagin is 1:5, and the DSPE-mPEG 2000 The molar ratio of the long-acting fragment to lecithin is 1:300, and the DSPE-PEG 2000 -FA targeting fragment and lecithin have a molar ratio of 0.2:50.

[0026] The preparation method of the pharmaceutical composition specifically comprises the following steps: S1: Weigh lecithin, cholesterol, docetaxel, plumbagin and DSPE-mPEG 2000 The long-acting fragment is dissolved in chloroform, mixed at normal pressure for 4 min, and after the drug and lecithin are fully mixed and homogenized, slowly evacuate to form a translucent film; S2: Add a physiological saline solution to the translucent film, hydrate the film by rotation at 37 °C for 2 h to obtain a mixed solution; centrifuge the mixed solution at 5000 rpm for 10 min, take the supernatant, and then add DSPE-PEG 2000 -FA targeting fragment to the supernatant and co-incubate at 37 °C for 2 h; S3: After the incubation is completed, the obtained solution is probed and ultrasonically treated in an ice-water bath for 3 s and cooled for 8 s, and the cycle operation is carried out for a total of 15 min, and ultrasonically treated in a water bath at room temperature for 10 min, and extruded through a membrane by a liposome extruder for 10 times to obtain a long-circulating targeted co-delivery liposome nanoparticle Lip-PEG-FA@DTX / PLB co-loading docetaxel and plumbagin, that is, the pharmaceutical composition.

[0027] Example 3 This example provides a pharmaceutical composition for radiosensitization in the treatment of non-small cell lung cancer. The pharmaceutical composition is composed of lecithin (egg yolk lecithin), cholesterol, docetaxel, plumbagin, DSPE-mPEG 2000 long-acting fragment and DSPE-PEG 2000 -FA targeting fragment composition.

[0028] The molar ratio of the above-mentioned lecithin to cholesterol is 10:1, the molar ratio of docetaxel to lecithin is 1:50, the molar ratio of docetaxel to plumbagin is 10:1, and the DSPE-mPEG 2000 The molar ratio of the long-acting fragment to lecithin is 1:500, and the DSPE-PEG 2000 -FA targeting fragment and lecithin have a molar ratio of 0.2:2.

[0029] The preparation method of the pharmaceutical composition specifically comprises the following steps: S1: Weigh lecithin, cholesterol, docetaxel, plumbagin, and DSPE-mPEG 2000 The long-acting fragment is dissolved in chloroform, mixed at normal pressure for 5 min. After the drug and lecithin are fully and evenly mixed, slowly evacuate to form a semi-transparent film; S2: Add physiological saline solution to the semi-transparent film, hydrate the film by rotation at 37 °C for 2 h to obtain a mixed solution; centrifuge the mixed solution at 5000 rpm for 10 min, take the supernatant, and then add the DSPE-PEG 2000 -FA targeting fragment to the supernatant and co-incubate at 37 °C for 2 h; S3: After the incubation, subject the obtained solution to probe sonication in an ice-water bath for 3 s and cool for 8 s, perform the cyclic operation for a total of 15 min, and perform water bath sonication at room temperature for 10 min. Extrude through the membrane of a liposome extruder repeatedly for 10 times to obtain the long-circulating targeted co-delivery liposome nanoparticles Lip-PEG-FA@DTX / PLB co-loaded with docetaxel and plumbagin, that is, the pharmaceutical composition.

[0030] Experimental Example 1 Characterization of the dual-loaded long-circulating targeted liposome nanoparticles Lip-PEG-FA@DTX / PLB The long-circulating targeted co-delivery liposome nanoparticles Lip-PEG-FA@DTX / PLB co-loaded with docetaxel and plumbagin prepared in Example 2 were tested as follows: 1. Observation of nanoparticle morphology Use a FEI Tecnai Spirit Biotwin 120 KV transmission electron microscope to observe the nanoparticle sample. The results are as Figure 1 shown. The nanoparticles are spherical-like and have uniform sizes, with a particle size of about 160 nm.

[0031] 2. Determination of nanoparticle size, PDI value, and Zeta potential Disperse the Lip-PEG-FA@DTX / PLB nanoparticles in ultrapure aqueous solution, and use a Malvern particle size analyzer to measure their size, PDI value, and Zeta potential. The results are shown in Table 1. The size of the obtained dual-loaded long-circulating nanoparticles is less than 200 nm, and the PDI is less than 0.2, indicating uniform sizes, and the zeta potential is -15.19 mV.

[0032] Table 1 Particle size, PDI value, and zeta potential of the Lip-PEG-FA@DTX / PLB nanoparticles of the present invention (n = 3)

[0033] 3. Determination of nanoparticle encapsulation efficiency and drug loading The co-loaded liposome Lip-PEG-FA@DTX / PLB was demulsified, and the encapsulation efficiency (EE) and drug loading content (LC) of docetaxel and plumbagin were determined by HPLC method. The results are shown in Table 2. The results showed that the nanoparticles could effectively encapsulate docetaxel and plumbagin, and the encapsulation effects of the two were similar, ensuring the co-delivery of DTX and PLB in proportion.

[0034] Table 2 Encapsulation efficiency and drug loading content of the present invention (n = 3)

[0035] 4. Detection of nanoparticle stability Storage stability: The Lip-PEG-FA@DTX / PLB liposome nanoparticles were stored at 4 °C for 7 days, and a Malvern particle size analyzer was used to detect the changes in their particle size, PDI value, and zeta potential. The experimental results are shown in Table 3. The results showed that after 7 days of storage, the particle size and potential did not change significantly, indicating that the nanoparticles could be stably stored at 4 °C.

[0036] Table 3 Particle size, PDI value, and zeta potential of the present invention (n = 3)

[0037] Dispersion stability: The liposomes were placed in a serum medium, and the change in encapsulation efficiency within 48 h was measured. Serum is the most commonly used release medium in in vitro release experiments. The results are as Figure 2 shown. The two drugs encapsulated in the liposomes had excellent stability in the serum medium.

[0038] 5. Investigation of in vitro release efficiency of nanoparticles The liposomes were placed in a dialysis bag, and the dialysis bag was immersed in release media of PBS (pH 7.4) buffer solution and acetate (pH 5.5) buffer solution. It was shaken at 37 °C and 100 rpm on a thermostatic shaker for 24 h. At predetermined time intervals, samples were taken to detect the drug concentration and a cumulative release curve was plotted. The results are as Figure 3 shown. Figure 3A is the release rate curve of DTX in the present invention within 24 h, and Figure 3B is the release rate curve of PLB in the present invention within 24 h. The results showed that compared with the neutral (pH 7.4) condition, docetaxel and plumbagin could be released from the nanoparticles more rapidly in the acidic environment of pH 5.5, which was beneficial for the rapid release of the nanoparticles in the acidic microenvironment of tumors. In addition, from Figure 3 the comparison of the results of Figures 3A and 3B, the release behaviors of docetaxel and plumbagin were similar, ensuring the synchronous release of the drugs in vivo.

[0039] 6. Implementation of in vitro cytotoxicity and radiosensitization of nanoparticles The human non-small cell lung cancer cell line A549 cells in the logarithmic growth phase were digested with trypsin (cultured in high-glucose DMEM medium containing 10% fetal bovine serum (4.5 g / L D-glucose, L-glutamine, 110 mg / L sodium pyruvate, phenol red), and 1% streptomycin and penicillin mixed solution was added, and placed in a 25 cm 2 culture flask, and cultured in an incubator at 37 °C and 5% CO 2 ). After digestion, centrifuged at 1000 rpm for 5 min, resuspended with the medium, and diluted by an appropriate multiple to make the cell concentration 5×10 4 cells / mL. Take 100 μL of the diluted cell suspension (5×10 3 cells / well) and inoculate it into a 96-well cell culture plate. At the same time, set control wells, blank wells, and drug-added wells. The drug-added wells are divided into a free drug group (DTX+PLB), a non-targeted nanoparticle group (Lip-PEG@DTX / PLB), and a targeted nanoparticle group (Lip-PEG-FA@DTX / PLB). When the cells in the wells grow to 80%, add drugs to 3 replicate wells, and add drugs at different concentrations (1, 2, 4, 8, 12, 15, 20, 25, 30, 40, 50 μmol / L) respectively (no drug is added to the blank well, PBS is added to the control well, DTX+PLB is added to the free drug group, Lip-PEG@DTX / PLB is added to the non-targeted nanoparticle group, and Lip-PEG-FA@DTX / PLB is added to the targeted nanoparticle group), incubate at 5% CO 2 , 37 °C for 24 h, then irradiate with 2 Gy X-rays and culture for another 24 h, and collect the cells. Add 10 μL of CCK-8 detection working solution, incubate for another 2 h, and then detect with an enzyme-linked immunosorbent assay at a wavelength of 450 nm. Calculate the relative cell survival rate and calculate the IC 50 .

[0040] The results are as Figure 4 shown. Figure 4A shows the cytotoxicity comparison of the free drug group (DTX+PLB), the non-targeted nanoparticle group (Lip-PEG@DTX / PLB), and the targeted nanoparticle group (Lip-PEG-FA@DTX / PLB) at the same drug concentration (10 μmol / L). The results show that the targeted nanoparticles have the strongest inhibitory effect on the growth of A549. Figure 4 Figures 4B and 4C are the cytotoxicity concentration-dependent curves of the three dosage forms and their corresponding IC 50 values. All three groups showed a dose-dependent cytotoxic effect on A549 cells. Among them, the Lip-PEG-FA@DTX / PLB nanoparticles showed the strongest killing effect on tumor cells and had the lowest IC 50 value, which was 5.97 μM. Figure 4D shows the cytotoxicity effect on A549 cells after Lip-PEG-FA@DTX / PLB nanoparticles combined with radiotherapy (RT). The results prove that after combined RT, the inhibitory effect on NSCLC is significantly enhanced, and the Lip-PEG-FA@DTX / PLB+RT group shows the strongest tumor inhibitory effect.

[0041] In some embodiments of the present invention, the long-circulating targeted co-delivery liposome nanoparticles Lip-PEG-FA@DTX / PLB prepared in Example 1 and Example 3 and co-loaded with docetaxel and plumbagin were tested in the above Experimental Example 1, and the same experimental results as those of the liposomes in Example 2 were obtained.

[0042] The liposome of this embodiment is used as a drug for treating lung cancer, and the DSPE-mPEG in it 2000 The long-acting fragment reduces the capture interference of liposomes with albumin and the reticuloendothelial system in vivo and prolongs the circulation time of liposomes in vivo; the DSPE-PEG in this liposome 2000 -FA targeting fragment, as a ligand of the folate receptor highly expressed on the surface of lung cancer cells, specifically recognizes and binds to the folate receptor on the surface of A549 cells, increasing the intracellular uptake of liposomes; this liposome promotes the production of reactive oxygen species, destroys the mitochondrial membrane potential, induces apoptosis, and increases the sensitivity to radiotherapy.

[0043] In summary, this novel co-loaded nano-drug delivery system will make important contributions to the academic development in the field of pharmaceutical preparations, provide new and more effective treatment methods for various malignant tumors clinically, and provide more important experimental evidence for improving the therapeutic effect of clinical radiotherapy combined with chemotherapy drugs, reducing their toxic side effects, and improving the quality of life of clinical cancer patients, having important theoretical value and good application prospects.

[0044] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A pharmaceutical composition for radiosensitization of non-small cell lung cancer, characterized in that: The pharmaceutical composition comprises lecithin, cholesterol, docetaxel, plumbagin, DSPE-mPEG 2000 Long-acting fragments and DSPE-PEG 2000 -FA targeting fragment composition.

2. A pharmaceutical composition for radiosensitization of non-small cell lung cancer according to claim 1, characterized in that: The lecithin is egg yolk lecithin.

3. A pharmaceutical composition for radiosensitization of non-small cell lung cancer according to claim 1, characterized in that: The molar ratio of lecithin to cholesterol is 2-10:1, the molar ratio of docetaxel to lecithin is 1:10-50, the molar ratio of docetaxel to plumbagin is 1:10-10:1, and the DSPE-mPEG 2000 The molar ratio of the long-acting fragment to lecithin is 1:100-500, and the DSPE-PEG 2000 -FA targeting fragment and lecithin molar ratio was 0.2:100~2.

4. A method for preparing a pharmaceutical composition for radiosensitization of non-small cell lung cancer according to any one of claims 1 to 3, characterized in that: The specific steps include: S1: Weigh lecithin, cholesterol, docetaxel, plumbagin and DSPE-mPEG 2000 The long-acting fragment is dissolved in chloroform and mixed at normal pressure for 3-5 minutes to allow the drug and lecithin to be fully mixed, and then slowly evacuated to form a translucent film; S2: Add physiological saline solution to the translucent film, rotate and hydrate the film at 37°C for 2 hours to obtain a mixed solution; centrifuge the mixed solution for 10 minutes, take the supernatant, and then 2000 -FA targeting fragments were added to the supernatant and incubated at 37°C for 2 h; S3: After the incubation, the resulting liquid was subjected to probe ultrasonication for 3s and cooled for 8s in an ice water bath, and the cycle was repeated for 15min, and then subjected to water bath ultrasonication for 10min at room temperature. The liquid was repeatedly extruded through the membrane through a liposome extruder for 10 times to obtain long-circulating targeted co-delivery liposome nanoparticles Lip-PEG-FA@DTX / PLB co-loaded with docetaxel and plumbagin, i.e., the pharmaceutical composition.

5. The method for preparing a pharmaceutical composition for radiosensitization of non-small cell lung cancer according to claim 4, characterized in that: In the step S1, the molar ratio of lecithin to cholesterol is 2-10:1, the molar ratio of docetaxel to lecithin is 1:10-50, the molar ratio of docetaxel to plumbagin is 1:10-10:1, and the DSPE-mPEG 2000 The molar ratio of the long-acting fragment to lecithin is 1:100-500.

6. The method for preparing a pharmaceutical composition for radiosensitization of non-small cell lung cancer according to claim 4, characterized in that: In step S2, DSPE-PEG 2000 The molar ratio of -FA targeting fragment to lecithin in step S1 was 0.2:100~2.

7. The method for preparing a pharmaceutical composition for radiosensitization of non-small cell lung cancer according to claim 4, characterized in that: In step S2, the centrifugal speed is 5000 rpm.

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