Preparation of multifunctional carrier-free nano-drug and drug combination application of multifunctional carrier-free nano-drug

Abstract

The invention provides preparation of a multifunctional carrier-free nano-drug and drug combination application of the multifunctional carrier-free nano-drug, and belongs to the technical field of medicines. The nano-drug comprises a copolymer material as well as a photosensitizer and docetaxel which are encapsulated in the copolymer material; wherein the mass ratio of the copolymer material to the photosensitizer to the docetaxel is (0.5 to 2): (0.075 to 0.3): (0.075 to 0.3). The drug docetaxel and the photosensitizer are encapsulated in the copolymer material, so that co-delivery of multiple drugs in vivo is realized, the tumor treatment effect can be effectively improved, the targeting property is excellent, and the biocompatibility is good.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to the preparation of a multifunctional carrier-free nano drug and its combined drug application. Background Art

[0002] At present, traditional methods of treating cancer mainly include surgical resection, radiotherapy, chemotherapy, etc. Surgical resection is to directly remove tumor tissue. It is generally used to treat solid tumors clinically and has significant therapeutic effects. However, for malignant and diffuse tumors, simple surgical resection cannot achieve complete resection and is very likely to recur. Radiotherapy refers to a treatment method that uses various types of radiation to irradiate tumors to kill or inhibit over-proliferation of cancer cells. However, the effect of radiation on killing cancer cells is affected by tissue sensitivity and tumor oxygen levels, and radiotherapy is not suitable when the tumor itself is insensitive to chemotherapy. Chemotherapy refers to the use of chemical drugs to enter the blood and tumor tissue to kill tumor cells. However, traditional chemotherapy drugs have problems such as high toxicity, poor selectivity, and strong side effects. The above problems are currently the main cause of death in chemotherapy patients.

[0003] Docetaxel (DTX), also known as docetaxel, is a paclitaxel derivative obtained by semi-synthesis of yew extract compounds; the chemical structure of DTX is shown below:

[0004]

[0005] The mechanism of action of DTX is to inhibit microtubule depolymerization while promoting microtubule formation. This mechanism causes cells to be unable to form spindle fibers and spindles during the interphase of mitosis, thereby blocking the cell cycle and inhibiting cell proliferation. Currently, DTX has been widely used in drugs for the treatment of various tumors such as metastatic breast cancer, ovarian cancer, small cell lung cancer, head and neck cancer, and prostate cancer.

[0006] Docetaxel is highly lipophilic, and its oral bioavailability is only 8%. The commercially available docetaxel injection is called Dopafil or Taxotere. Before clinical use, it must be dissolved in a special solvent of 13% ethanol-water solution, and then diluted with 5% glucose injection or 0.9% NaCl solution. Tween 80 is used as a solubilizer in the injection to increase cell membrane fluidity and membrane permeability, but Tween 80 often leads to severe hypersensitivity reactions, fluid retention, hemolysis, cholestasis and other adverse reactions. Therefore, patients need to take glucocorticoid drugs in advance in clinical practice. In addition, Tween 80 has a certain degree of interference with the in vivo pharmacokinetic behavior of DTX, making it nonlinear pharmacokinetic. Ethanol is used as a cosolvent, and its excessive intake may cause adverse reactions such as memory loss and body numbness. The above adverse reactions seriously limit the clinical and drug preparation applications of docetaxel. Therefore, improving the targeting of docetaxel is an important means to improve its efficacy.

[0007] Photodynamic therapy (PDT) has attracted widespread attention because it can selectively treat tumor lesions locally and precisely control the treatment site. PDT is a method in which photosensitizers are accumulated in tumor tissues and then the tumor site is irradiated with light of a specific wavelength. After the photosensitizers absorb the energy of photons, some of their electrons are excited. The excited photosensitizers transfer energy to oxygen, thereby generating some reactive oxygen species (ROS). ROS attack cell structures through oxidation, causing oxidative damage to cell membranes or proteins. When the accumulation of oxidative damage reaches a certain level, it can kill cancer cells.

[0008] Mixed polymer micelles have been widely studied as effective carriers for low water-soluble drugs. By site-specific administration, the biopharmaceutics and pharmacokinetic properties of drugs can be improved, thereby improving the efficacy of drugs. Although traditional microparticles or nanoparticles are formed by complex emulsification processes and require the use of surfactants for stabilization, mixed polymer micelles with a particle size less than 1 micron are usually prepared by a simplified self-assembly process and do not require surfactants or other agents to achieve stability. At present, mixed polymer micelles are formed by various types of polymers, including copolymers and lipid polymers. There are also relevant reports in the following prior art: Lee et al., Journal of Controlled Release, 2003, 91, 103-113 (Lee et al., published in Journal of Controlled Release Vol. 91, pp. 103-113 in 2003) and Vakil et al., Langmuir, 2006, 22, 9723-9729 (Vakil et al. published in Langmuir Vol. 22, pp. 9723-9729 in 2006). Compositions containing these polymer micelles still need to be improved in terms of enhancing targeting and biocompatibility.

[0009] Therefore, there is an urgent need for a polymer micelle that can be used for photodynamic therapy. Summary of the invention

[0010] The purpose of the present invention is to provide a preparation method of a multifunctional carrier-free nano drug and its combined drug application, which can effectively improve the tumor treatment effect, has excellent targeting and good biocompatibility.

[0011] To achieve the above-mentioned purpose of the invention, the technical solution of the present invention is as follows:

[0012] In a first aspect of an embodiment of the present invention, a multifunctional carrier-free nanomedicine is provided, comprising a copolymer material and a photosensitizer and docetaxel encapsulated in the copolymer material; wherein the mass ratio of the copolymer material, the photosensitizer and docetaxel is 0.5-2:0.075-0.3:0.075-0.3.

[0013] In one aspect of the present invention, the average particle size of the multifunctional carrier-free nanomedicine is 180nm-220nm.

[0014] In one aspect of the present invention, the copolymer material is block polyether F-127.

[0015] In one aspect of the present invention, the photosensitizer is chlorin Ce6.

[0016] In one aspect of the present invention, the mass ratio of the copolymer material, the photosensitizer and docetaxel is 0.5-2:0.075-0.3:0.075-0.3.

[0017] In one aspect of the present invention, the mass ratio of the copolymer material, the photosensitizer and docetaxel is 0.8-1.2:0.125-0.18:0.125-0.18.

[0018] In one aspect of the present invention, the mass ratio of the copolymer material, the photosensitizer and docetaxel is 1:0.15:0.15.

[0019] In a second aspect of the embodiment of the present invention, a method for preparing the above-mentioned multifunctional carrier-free nanomedicine is provided, comprising the following steps:

[0020] Step 1: dissolving the copolymer material, the photosensitizer and docetaxel in an organic solvent, and evaporating the solvent at 40° C. to 50° C. to form a film;

[0021] Step 2: Add water to the product obtained in step 1 to obtain a micellar aqueous solution after complete dissolution;

[0022] Step 3: adding a first solution to the micellar aqueous solution, and then stirring to obtain a solution; the first solution is selected from a mixed solution of MPTMS and ammonia water;

[0023] Step 4: Collect the solution obtained in step 3, and dialyze the solution to obtain a multifunctional carrier-free nanomedicine after separation.

[0024] In one aspect of the present invention, the organic solvent is selected from at least one of anhydrous ethanol, acetone, propanol, ethyl acetate and ethyl formate; more preferably anhydrous ethanol.

[0025] In one aspect of the present invention, in the first solution, the volume ratio of MPTMS to aqueous ammonia is 6-7:0.8-1.2.

[0026] In one aspect of the present invention, in the first solution, the volume ratio of MPTMS to aqueous ammonia is 6.25:1.

[0027] In one aspect of the invention, the method comprises the following steps:

[0028] Step 1: dissolving the block polyether F-127, dihydrochlorin Ce6 and docetaxel in anhydrous ethanol, and evaporating the mixture into a film by a rotary evaporator at 40° C.-50° C., wherein the mass ratio of the block polyether F-127, dihydrochlorin Ce6 and docetaxel is 0.8-1.2:0.125-0.18:0.125-0.18;

[0029] Step 2: Add water to the product obtained in step 1, and rotate at a low speed at 40°C-50°C until it is completely dissolved to obtain a micellar aqueous solution;

[0030] Step 3: adding a mixed solution of MPTMS and aqueous ammonia to the micellar aqueous solution, wherein the volume ratio of the MPTMS to aqueous ammonia is 6-7:0.8-1.2, and then stirring overnight to obtain a solution;

[0031] Step 4: Collect the solution obtained in step 3, dialyze the solution, and obtain a multifunctional carrier-free nanomedicine after centrifugation.

[0032] In a third aspect of the embodiments of the present invention, there is provided application of the aforementioned multifunctional carrier-free nanomedicine in the preparation of anti-tumor drugs.

[0033] Preferably, the tumor is selected from at least one of non-small cell lung cancer, colon cancer, breast cancer, gastric cancer, esophageal cancer, prostate cancer, leukemia, head and neck cancer, pancreatic cancer, multiple myeloma or liver cancer.

[0034] More preferably, the tumor is breast cancer.

[0035] Beneficial Effects of the Invention

[0036] The present invention improves the limitation of single drug treatment and provides a multifunctional carrier-free nano drug which can effectively improve the tumor treatment effect, has excellent targeting and good biocompatibility. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 Schematic diagram of the preparation principle of multifunctional carrier-free nanomedicine.

[0038] Figure 2A The pictures were taken under a laser microscope after 4T1 cells were treated with different concentrations of Ce6 and FDC, scale bar = 20 μm.

[0039] Figure 2B The statistical results of laser microscope fluorescence values ​​after 4T1 cells were treated with different concentrations of Ce6 and FDC.

[0040] Figure 2C The pictures were taken under a laser microscope after 4T1 cells were treated with the same concentration of Ce6 and FDC for different incubation times. Scale bar = 20 μm.

[0041] Figure 2D The statistical results of laser microscope fluorescence values ​​after 4T1 cells were treated with the same concentration of Ce6 and FDC at different incubation times.

[0042] Figure 2EThe statistical results of fluorescence values ​​measured by flow cytometry after 4T1 cells were treated with different concentrations of Ce6 and FDC.

[0043] Figure 2F The fluorescence value statistics are obtained by flow cytometry after 4T1 cells were treated with the same concentration of Ce6 and FDC at different incubation times.

[0044] Figure 3 In Example 1, the SOSG method was used to detect the singlet oxygen pattern in vitro using fluorescence spectroscopy.

[0045] Figure 4A This is a laser microscope photo of 4T1 cells treated with the nanodrug sample of Example 1 and incubated with the DCFH-DA probe. Scale bar = 20 μm.

[0046] Figure 4B These are the statistical results of laser microscope fluorescence values ​​after 4T1 cells were treated with the nanodrug sample of Example 1 and incubated with the DCFH-DA probe.

[0047] Figure 5 is the MTT assay diagram of Example 1; Figure 5 (a), the cell survival rates of each concentration gradient from left to right are Ce6, DTX, Ce6+DTX and FDC; Figure 5 In (b), the cell viability of each concentration gradient from left to right is Ce6(+), Ce6+DTX(+) and FDC(+).

[0048] Figure 6 This is a cell staining test diagram of Example 1 of the present invention.

[0049] Fig. 7A This is a graph showing apoptosis tests of the blank group, Ce6, DTX and Ce6(+) in Example 1 of the present invention.

[0050] Figure 7B This is a graph showing the cell apoptosis test of Ce6+DTX, FDC, Ce6+DTX(+), and FDC(+) in Example 1 of the present invention. DETAILED DESCRIPTION

[0051] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is only an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the invention of the present invention based on the disclosed content, and they should also fall within the scope of the present application.

[0052] In the following examples, the block polyether F-127 was purchased from Beijing Solebow Technology Co., Ltd., with the product number GS2975;

[0053] The photosensitizer chlorin Ce6 was purchased from Frontier Scientific, Inc., with the catalog number P562-9;

[0054] Docetaxel was purchased from MedChemExpress with the product number HY-B0011.

[0055] The above purchase information is only used as an example of a specific embodiment and does not impose any substantial limitation on the present invention.

[0056] In the following embodiments,

[0057] “FDC” is a nano drug formed by self-assembly of three drugs: photosensitizer (Ce6) + docetaxel (DTX) and block polyether F-127 (Pluronic F-127) in the present invention;

[0058] “MPTMS” is (3-mercaptopropyl)trimethoxysilane, CAS number: 4420-74-0.

[0059] In the present invention, a method for preparing a multifunctional carrier-free nanomedicine is provided, the method comprising the following steps:

[0060] Step 1: providing a block polyether F-127, a photosensitizer dihydrochlorin Ce6 and docetaxel; dissolving the block polyether F-127, the photosensitizer dihydrochlorin Ce6 and docetaxel in an organic solvent, and evaporating at 40° C.-50° C. to form a film;

[0061] Step 2: Add water to the product obtained in step 1 to obtain a micellar aqueous solution after complete dissolution;

[0062] Step 3: adding the first solution to the micellar aqueous solution, and then stirring to obtain a solution;

[0063] Step 4: Collect the solution obtained in step 3, dialyze the solution, and obtain a multifunctional carrier-free nanomedicine after centrifugation.

[0064] The preparation principle of the present invention is as follows Figure 1 As shown, the photosensitizer dihydrochlorin Ce6 and docetaxel are encapsulated in the block polyether F-127 by thin film hydration method to obtain FDC, which is the multifunctional carrier-free nano drug used in the present invention.

[0065] The present invention is further described below by way of specific examples. The various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the contents described below are all weight contents. Unless otherwise specified, it is understood that the process is carried out at room temperature.

[0066] Example 1

[0067] The multifunctional carrier-free nanomedicine of Example 1 was prepared by thin film hydration method: a mixture of 5 mg of block polyether F-127, 0.75 mg of DTX and 0.75 mg of Ce6 was completely dissolved in 5 mL of anhydrous ethanol, placed in a round-bottom bottle, and evaporated into a film by a rotary evaporator at a water temperature of 45°C. Then 2 mL of H2O was added and rotated at a low speed of 45°C until completely dissolved to obtain a micellar aqueous solution. Then 12.5 μL of (3-mercaptopropyl) trimethoxysilane (MPTMS) and 2 μL of ammonia solution were added and stirred overnight. The above solution was collected and treated with ultrapure water membrane dialysis (MWCO: 8000-14000Da) for 3 hours to remove impurities. After centrifugation at 14000rpm for 15 minutes, the supernatant was retained to obtain the nanomaterial of Example 1, which was stored at 4°C.

[0068] The grouping used in the cell experiment of the present invention is:

[0069] Ce6: dark treatment group with photosensitizer Ce6 only (without laser irradiation);

[0070] DTX: docetaxel alone;

[0071] C+D: docetaxel alone + photosensitizer Ce6 alone dark treatment group (no laser irradiation treatment);

[0072] FDC: photosensitizer (Ce6) + docetaxel (DTX) + block polyether F-127 (F127) three drugs self-assemble to form a nano drug dark treatment group, that is, the dark treatment group of Example 1 (no laser light treatment);

[0073] Ce6(+): group treated with photosensitizer Ce6 alone;

[0074] C+D(+): docetaxel alone + photosensitizer Ce6 alone light treatment group;

[0075] FDC(+): Photosensitizer (Ce6) + docetaxel (DTX) block polyether F-127 (Pluronic F127) three drugs self-assemble to form a nano drug light-treated group, that is, the light-treated group of Example 1.

[0076] Experiment 1: Characterization of Nanoparticles

[0077] The drug particle size, PDI and zeta potential were detected using a Malvern laser particle size analyzer FDC. Ce6 and DTX in FDC nanomaterials were quantified using high performance liquid chromatography (HPLC), and their encapsulation efficiency (EE) and drug loading efficiency (DLE) were calculated based on the standard curve of free drugs; where: EE = drug mass / actual dose, DLE = drug mass / total mass of nanoparticles. The characterization results are shown in Table 1 below:

[0078] Table 1

[0079]

[0080] Experiment 2: In vitro cell experiments

[0081] 1. Cell culture:

[0082] Mouse breast cancer cells (4T1) were cultured with 10% fetal bovine serum and 1% antibiotics at 37°C in a humidified atmosphere containing 5% CO2.

[0083] 2. In vitro ROS detection:

[0084] The SOSG method was used to detect singlet oxygen in vitro by fluorescence spectroscopy. That is, singlet oxygen green fluorescent probe SOSG (5 μM) was incubated with FDC (3.65 mg / L) or the same concentration of Ce6 (2 mg / L) and DTX (1.65 mg / L) in PBS; regardless of the presence or absence of light irradiation, the SOSG fluorescence of the mixed solution was measured every 15 seconds to detect the generation of ROS. The results are shown in Figure 2. Figure 3 As shown; Figure 3 The values ​​of PBS, F127, DTX, Ce6 and FDC are similar.

[0085] In addition, 4T1 cells were inoculated and cultured for 24 hours, treated with FDC (9.12 mg / L) or the same concentration of Ce6 (5 mg / L) and DTX (4.12 mg / L) for 6 hours, washed with PBS three times, and incubated with DCFH-DA (10 μM), a probe for detecting intracellular reactive oxygen species (ROS), for 20 minutes. For the light group, the cells were exposed to light for 5 minutes; for the dark group, the cells were cultured in a light mask. Subsequently, the cells were washed with PBS and the intracellular fluorescence was observed using CLSM. The results are shown in Figure 2. Figure 4A and Figure 4B shown.

[0086] 3. Cellular uptake:

[0087] CLSM was used to detect cell uptake, and flow cytometry was used to detect 4T1 cells inoculated and cultured for 24 hours. After removing the culture medium, the cells were treated with free Ce6 and FDC (using Ce6 2 / 4 / 8 mg / L concentrations, respectively) for 8 hours, and with Ce6 4 mg / L for 2 / 4 / 8 hours. Then, the cells were washed with PBS, 1× Hoechst 33342 live cell staining solution was added, and the cells were incubated for 20-30 minutes in the dark. After washing with PBS three times, CLSM was used to observe the endocytosis, as shown in Figure 2. Figure 2A , Figure 2B , Figure 2C and Figure 2D . Flow cytometry: 4T1 cells were inoculated in a 6-well plate for 24 hours, treated according to the above concentration and time gradient, and then washed once with PBS. 500μM trypsin was added to digest the cells for 2-3 minutes; trypsin was aspirated, and 500μM DMEM was added to collect the cells into an EP tube. After centrifugation at 800rpm for 3 minutes, the supernatant was removed. The precipitate was suspended with 200μl PBS and stored on ice; flow cytometry was used to detect the cell fluorescence characteristics and evaluate the drug uptake. The results are shown in the figure. Figure 2E and Figure 2F (Concentration is 4 mg / L).

[0088] 4.MTT assay:

[0089] 4T1 cells were seeded in a 96-well plate, cultured for 24 hours, and incubated with gradient concentrations of drugs (Ce6, DTX, Ce6+DTX, and FDC) for 6 hours. For the light group, the cells were exposed to light for 5 minutes. For the dark group, the cells were cultured in the light mask. After 24 hours, 20 μL MTT (dissolved in 5g / L ultrapure water) was added to each well for 4 hours. Crystal violet was dissolved in 150 μL DMSO, and the absorbance value of each well was detected with a 570nm microplate reader. The results are shown in Figure 2. Figure 5 shown.

[0090] 5. Live / dead cell staining:

[0091] 4T1 cells were inoculated and cultured for 24 hours, and then treated with FDC (14.58 mg / L), Ce6 (8 mg / L), DTX (6.58 mg / L), and Ce6+DTX for 6 hours. For the light group, the cells were irradiated with light for 15 minutes; for the dark group, the cells were cultured in the light mask all the time, and then all the cells were cultured for another 3 hours. The cells were stained with Calcein-AM / PI, a live cell / dead cell double staining kit, for 20 minutes, washed twice with PBS, and then observed with CLSM. The results are shown in Figure 2. Figure 6 shown.

[0092] 6. Cell apoptosis assay:

[0093] 4T1 cells were inoculated in 6-well plates for 24 hours and incubated with the test samples of each group in Example 1 for 6 hours:

[0094] FDC (containing Ce6: 2 mg / L, DTX: 1.65 mg / L), Ce6 (2 mg / L), DTX (1.65 mg / L), Ce6+DTX (containing Ce6: 2 mg / L, DTX: 1.65 mg / L).

[0095] The light treatment group received light irradiation for 90 seconds, and the dark treatment group received dark irradiation. Subsequently, the 4T1 cells were collected and resuspended in 100 μL of apoptosis binding solution. Then, the cells were stained with Annexin V-FITC and PI for 15 minutes; the results are shown in Fig. 7A and Figure 7B shown. Fig. 7A This is a graph showing apoptosis tests of the blank group, Ce6, DTX and Ce6(+) in Example 1 of the present invention. Figure 7B This is a graph showing the cell apoptosis test of Ce6+DTX, FDC, Ce6+DTX(+), and FDC(+) in Example 1 of the present invention.

[0096] Experiment 3: In vitro cytotoxicity assay

[0097] 1. Cell culture:

[0098] The aforementioned mouse breast cancer cells were used.

[0099] 2. In vitro cytotoxicity test:

[0100] After digestion, the breast tumor cell suspension was inoculated with 100 μL per well on a 96-well plate, and the breast tumor cell density was adjusted to 5×10 3 / well, the edge wells were filled with sterile PBS; incubated in a 37°C, 5% CO2 incubator for 24 hours until the cell monolayer covered the bottom of the well (96-well flat-bottom plate), and the multifunctional carrier-free nanomedicine prepared in Example 1 was added at a concentration of 50 μg / mL, and 6 replicate wells were set for each concentration gradient of the sample; the cell plate after drug addition was placed in a 37°C, 5% CO2 incubator for 48 hours and then the culture was terminated, 10 μL of CCK-8 reagent was added to each well, and then placed in a 37°C, 5% CO2 incubator for 1-4 hours; the cells were exposed to light for 5 minutes, the OD value of each well was measured at a wavelength of 450nm, and the average OD value was calculated, wherein: cell survival rate = (OD value of drug addition group - OD value of blank group) / (OD value of non-drug addition group - OD value of blank group) × 100%; the results of Example 1 are as follows Figure 5 .

[0101] The results show that the nanomedicine obtained by the preparation method provided by the present invention has a better therapeutic effect on breast tumor cells.

[0102] Comparative Example 1

[0103] A mixture of 5 mg of block polyether F-127, 0.75 mg of DTX and 0.75 mg of Ce6 was completely dissolved in 5 mL of anhydrous ethanol, sonicated for 0.5 h, and then stirred overnight at room temperature, the solution was collected, and treated with ultrapure water membrane dialysis (MWCO: 8000-14000 Da) for 4 h to remove impurities, and centrifuged at 14000 rpm for 15 min.

[0104] This preparation method cannot form nanomedicine.

[0105] Comparative Example 2

[0106] A mixture of 5 mg of block polyether F-127, 1 mg of DTX and 0.75 mg of Ce6 was completely dissolved in 5 mL of anhydrous ethanol, placed in a round-bottom bottle, and evaporated into a film by a rotary evaporator at a water temperature of 45°C. Then 2 mL of H2O was added and rotated at a low speed of 45°C until completely dissolved to obtain a micellar aqueous solution. Then 15 μL of toluenesulfonic acid solution was added and stirred overnight. The solution was collected and treated with ultrapure water membrane dialysis (MWCO: 8000-14000Da) for 2 hours to remove impurities. Centrifuged at 14000rpm for 15 minutes.

[0107] This preparation method cannot form nanomedicine.

[0108] Comparative Example 3

[0109] The difference from Example 1 is that the temperature of the preparation method is different:

[0110] A mixture of 5 mg of block polyether F-127, 0.75 mg of DTX and 0.75 mg of Ce6 was completely dissolved in 5 mL of anhydrous ethanol, placed in a round-bottom bottle, and evaporated into a film by a rotary evaporator at a water temperature of 80°C. Then 2 mL of H2O was added and rotated at a low speed of 60°C until completely dissolved to obtain a micellar aqueous solution. Then 12.5 μL of MPTMS and 2 μL of ammonia solution were added and stirred overnight. The solution was collected and treated with ultrapure water membrane dialysis (MWCO: 8000-14000Da) for 3 hours to remove impurities. Centrifuged at 14000rpm for 15 minutes.

[0111] This preparation method cannot form nanomedicine.

[0112] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A multifunctional carrier-free nanomedicine, characterized in that: The invention comprises a copolymer material and a photosensitizer and docetaxel encapsulated in the copolymer material; wherein the mass ratio of the copolymer material, the photosensitizer and the docetaxel is 0.5-2:0.075-0.3:0.075-0.

3.

2. The multifunctional carrier-free nanomedicine according to claim 1, characterized in that: The average particle size of the multifunctional carrier-free nano drug is 180nm-220nm.

3. The multifunctional carrier-free nanomedicine according to claim 1, characterized in that: The copolymer material is block polyether F-127.

4. The multifunctional carrier-free nanomedicine according to claim 1, characterized in that: The photosensitizer is dihydrochlorin Ce6.

5. The multifunctional carrier-free nanomedicine according to claim 1, characterized in that: The mass ratio of the copolymer material, the photosensitizer and docetaxel is 0.8-1.2:0.125-0.18:0.125-0.

18.

6. The multifunctional carrier-free nanomedicine according to claim 5, characterized in that: The mass ratio of the copolymer material, the photosensitizer and docetaxel is 1:0.15:0.

15.

7. The method for preparing the multifunctional carrier-free nanomedicine according to any one of claims 1 to 6, characterized in that: The following steps are involved: Step 1: dissolving the copolymer material, the photosensitizer and docetaxel in an organic solvent, and evaporating the solvent at 40° C. to 50° C. to form a film; Step 2: Add water to the product obtained in step 1 to obtain a micellar aqueous solution after complete dissolution; Step 3: adding a first solution to the micellar aqueous solution, and then stirring to obtain a solution; the first solution is a mixed solution of MPTMS and ammonia water; Step 4: Collect the solution obtained in step 3, and dialyze the solution to obtain a multifunctional carrier-free nanomedicine after separation.

8. The method according to claim 7, characterized in that In the first solution, the volume ratio of MPTMS to aqueous ammonia is 6-7:0.8-1.

2.

9. The method according to claim 7 or 8, characterized in that: The following steps are involved: Step 1: dissolving the block polyether F-127, dihydrochlorin Ce6 and docetaxel in anhydrous ethanol, and evaporating the mixture into a film by a rotary evaporator at 40° C.-50° C., wherein the mass ratio of the block polyether F-127, dihydrochlorin Ce6 and docetaxel is 0.8-1.2:0.125-0.18:0.125-0.18; Step 2: Add water to the product obtained in step 1, and rotate at a low speed at 40°C-50°C until it is completely dissolved to obtain a micellar aqueous solution; Step 3: adding a mixed solution of MPTMS and aqueous ammonia to the micellar aqueous solution, wherein the volume ratio of the MPTMS to aqueous ammonia is 6.25:1, and then stirring overnight to obtain a solution; Step 4: Collect the solution obtained in step 3, dialyze the solution, and obtain a multifunctional carrier-free nanomedicine after centrifugation.

10. Use of the multifunctional carrier-free nanomedicine according to any one of claims 1 to 6 in the preparation of anti-tumor drugs.

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