PSO / PTX targeted composite nanoparticles, and preparation method and application thereof

Abstract

The embodiment of the invention provides PSO / PTX targeted composite nanoparticles as well as a preparation method and application thereof. The preparation method comprises the following steps: dissolving a phospholipid composition in a first organic solvent, adding water and an emulsifier after dissolving, and uniformly mixing to obtain a water phase; the phospholipid composition comprises phospholipid and modified phospholipid, dissolving PLGA in a second organic solvent, adding PTX, PSO and SPIONs (at) OA after dissolving, and uniformly mixing to obtain an organic phase; and adding the organic phase into the water phase, carrying out ultrasonic emulsification, evaporating to remove the organic solvent, and filtering to obtain the PSO / PTX targeted composite nanoparticles. According to the PSO / PTX targeted composite nanoparticles prepared by the embodiment of the invention, the fact that psoralen and paclitaxel are co-loaded on the superparamagnetic polymer lipid nanoparticles is proposed for the first time, and the PSO / PTX targeted composite nanoparticles have the functions of magnetic targeting, long circulation and slow release, so that the PSO / PTX targeted composite nanoparticles have a good application prospect in preparation of the medicines for treating the breast cancer.

Description

Technical Field

[0001] The embodiments of the present application relate to the technical field of drug synthesis, and particularly to a PSO / PTX targeted composite nanoparticle, a preparation method thereof, and uses thereof. Background Art

[0002] Triple-Negative Breast Cancer (TNBC) is a special subtype of breast cancer, characterized by the lack of expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) in tumor cells. It is highly invasive and heterogeneous. Currently, chemotherapy is one of the main clinical treatment methods for TNBC, but it is prone to multi-drug resistance (MDR), and there is a lack of effective drug resistance reversal agents on the market.

[0003] In the prior art, most studies on tumor drug resistance mechanisms and reversal strategies are limited to the level of tumor cells themselves and population cells, ignoring the influence of the tumor microenvironment. Relevant studies have shown that immune cells such as tumor-associated macrophages and immune factors constitute the immune microenvironment, which is closely related to the development of TNBC.

[0004] Psoralen (PSO) has immunomodulatory and anti-tumor effects. Its combination with the chemotherapeutic drug paclitaxel (PTX) can inhibit the growth of TNBC and reverse breast cancer MDR. However, the water solubility, stability, and targeting of PSO and PTX are poor, which restricts the practical application of PSO and PTX.

[0005] Therefore, how to improve the defects existing in the use of PSO / PTX is of great significance for optimizing the treatment strategy of TNBC. Summary of the Invention

[0006] The purpose of the embodiments of the present application is to provide a PSO / PTX targeted composite nanoparticle, a preparation method thereof, and uses thereof.

[0007] To achieve the above purpose, the embodiments of the present application propose the following technical solutions:

[0008] In a first aspect, the embodiments of the present application provide a preparation method of a PSO / PTX targeted composite nanoparticle, and the preparation method includes:

[0009] Dissolve a phospholipid composition in a first organic solvent. After dissolution, add water and an emulsifier, and mix evenly to obtain an aqueous phase; wherein, the phospholipid composition includes phospholipids and modified phospholipids.

[0010] Dissolve PLGA in a second organic solvent. After dissolution, add PTX, PSO, and SPIONs@OA, and mix evenly to obtain an organic phase.

[0011] The organic phase was added to the aqueous phase, and after ultrasonic emulsification, evaporation to remove the organic solvent, and filtration, PSO / PTX targeted composite nanoparticles were obtained.

[0012] As an embodiment, the preparation method of the SPIONs@OA includes:

[0013] Under an inert atmosphere, FeCl2·4H2O and FeCl3·6H2O were taken, added to ultrapure water, and mixed evenly to obtain solution A;

[0014] Ultrapure water was added to ammonia water and mixed evenly to obtain solution B;

[0015] Sodium oleate was taken and added to ultrapure water, and after ultrasonic dissolution, solution C was obtained;

[0016] Under an ice-water bath and stirring, solution B was added to solution A to obtain a mixed solution; the mixed solution was transferred to a water bath and stirred to prepare a SPIONs suspension;

[0017] The SPIONs suspension was separated by a magnet to obtain magnetic nanoparticles and supernatant;

[0018] Solution C was added to the magnetic nanoparticles, and the reaction was stirred under a water bath to obtain SPIONs@OA.

[0019] As an embodiment, the modified phospholipid is DSPE-PEG2000.

[0020] As an embodiment, the mass ratio of the phospholipid composition to PLGA is 1:1.

[0021] As an embodiment, at least one of the following conditions is satisfied:

[0022] The first organic solvent is anhydrous ethanol and / or methanol;

[0023] The second organic solvent is at least one of chloroform, dichloromethane or acetone;

[0024] The emulsifier is Tween.

[0025] As an embodiment, the mass ratio of the total mass of PTX and PSO to PLGA is 0.1:1.

[0026] As an embodiment, the adding of the organic phase to the aqueous phase, followed by ultrasonic emulsification, evaporation to remove the organic solvent, and filtration to obtain PSO / PTX targeted composite nanoparticles includes:

[0027] The organic phase was added to the aqueous phase, ultrasonic emulsification was carried out at an ultrasonic power of 225 W for 5 min, the organic solvent was evaporated at 45 °C for 5 min, and after filtration, PSO / PTX targeted composite nanoparticles were obtained.

[0028] In a second aspect, an embodiment of the present application provides a PSO / PTX targeted composite nanoparticle, which is prepared by the preparation method described in the first aspect.

[0029] In a third aspect, an embodiment of the present application provides the use of the PSO / PTX targeted composite nanoparticle described in the second aspect in the preparation of a drug for preventing, alleviating or treating breast cancer.

[0030] As an implementation manner, the drug is a drug for inhibiting the growth of MDA-MB-231 cells.

[0031] Compared with the prior art, the embodiment of the present application has at least the following beneficial effects:

[0032] The embodiment of the present application uses modified SPIONs (SPIONs@OA) to improve the delivery efficiency of PSO / PTX. Among them, in this embodiment, PTX, PSO and SPIONs@OA are coated by PLGA, and then PLGA is modified by phospholipids and modified phospholipids (forming an oil-in-water emulsion layer) to improve the encapsulation efficiency of PTX and PSO, so as to increase the drug loading amount and the stability of PSO and PTX. At the same time, the surface of the modified SPIONs is easy to be modified and functionalized, and the phospholipids and modified phospholipids can modify the surface of the SPIONs to improve the delivery efficiency of PSO and PTX. Therefore, the PSO / PTX targeted composite nanoparticle prepared in this embodiment first proposes to co-load psoralen (PSO) and paclitaxel (PTX) in superparamagnetic polymer lipid nanoparticles, solving the problem of dual-drug synergistic delivery. The PSO / PTX targeted composite nanoparticle has both magnetic targeting, long circulation and sustained release functions, and has high clinical transformation potential, indicating that the PSO / PTX targeted composite nanoparticle has good application prospects in the preparation of drugs for treating breast cancer.

[0033] Additional aspects and advantages of the present application will be given in part in the following description, and these will become apparent from the following description, or can be understood through the practice of the present application. Description of the Drawings

[0034] Figure 1 Shows the particle size response surface diagram of P / P-TCNs in Example 3;

[0035] Figure 2 Shows the encapsulation efficiency response surface diagram of PSO in P / P-TCNs in Example 3;

[0036] Figure 3 Shows the encapsulation efficiency response surface diagram of PTX in P / P-TCNs in Example 3;

[0037] Figure 4Shows the schematic diagram of the test results of infrared analysis in Example 6;

[0038] Figure 5 Shows the schematic diagram of the test results of cytotoxicity test in Example 7. Detailed implementation manners

[0039] Next, the technical solutions in the embodiments will be described clearly and completely in conjunction with the embodiments of the present application and the accompanying drawings. Obviously, the embodiments described below are only a part of the embodiments of the present application, rather than all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present application.

[0040] It should be understood that when used in this specification and the appended claims, the terms "comprises" and "comprising" indicate the presence of the described features, wholes, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, wholes, steps, operations, elements, components, and / or their combinations.

[0041] It should also be understood that the terms used in the specification of the embodiments of the present application are only for the purpose of describing specific embodiments and are not intended to limit the embodiments of the present application. As used in the specification of the embodiments of the present application and the appended claims, unless the context clearly indicates otherwise, the singular forms "a", "an", and "the" are intended to include the plural forms.

[0042] First, some terms and materials involved in this embodiment will be explained below to facilitate the understanding of those skilled in the art.

[0043] PSO: Psoralen.

[0044] PTX: Paclitaxel.

[0045] PEG-PE: DSPE-PEG2000, or modified phospholipid.

[0046] SPIONs: Superparamagnetic iron oxide nanoparticles, or magnetic nanoparticles.

[0047] SPIONs@OA: Superparamagnetic iron oxide nanoparticles coated with oleic acid (OA).

[0048] P / P-TCNs: Or PSO / PTX targeted composite nanoparticles, that is, targeted composite nanoparticles.

[0049] Unless otherwise specified, the water used in this embodiment is ultrapure water or deoxygenated ultrapure water.

[0050] Next, the technical solutions of the embodiments of the present application will be described in detail.

[0051] First, a preparation method of PSO / PTX targeted composite nanoparticles according to the first aspect of this embodiment will be described.

[0052] Preparation method of PSO / PTX targeted composite nanoparticles

[0053] In the prior art, due to the determined pharmacological activities and application significances of psoralen and paclitaxel, various nanoparticles prepared from psoralen and paclitaxel have been reported in many domestic and foreign studies, but each type of nanoparticle has its own different limitations. For example, psoralen (PSO) has immunomodulatory and antitumor effects, and its combination with the chemotherapeutic drug paclitaxel (PTX) can inhibit the growth of TNBC and reverse breast cancer MDR. However, the water solubility, stability, and targeting properties of PSO and PTX are poor, which restricts the practical applications of PSO and PTX.

[0054] In view of this, this embodiment proposes a preparation method of PSO / PTX targeted composite nanoparticles, and this preparation method includes the following steps:

[0055] (1) Dissolve the phospholipid composition in the first organic solvent, and after dissolution, add water and an emulsifier, and mix evenly to obtain an aqueous phase; wherein, the phospholipid composition includes phospholipids and modified phospholipids.

[0056] In this step, dissolve the phospholipids and modified phospholipids in the first organic solvent and the emulsifier, and the obtained aqueous phase can be used to modify (coat) PLGA subsequently to form an oil-in-water emulsion layer.

[0057] Preferably, the first organic solvent is absolute ethanol and / or methanol.

[0058] (2) Dissolve PLGA in the second organic solvent, and after dissolution, add PTX, PSO, and SPIONs@OA, and mix evenly to obtain an organic phase.

[0059] In step (2), first load PTX and PSO on SPIONs@OA, and then coat PTX, PSO, and SPIONs@OA with PLGA to form a core. In this step, SPIONs@OA realizes the loading of PTX and PSO, which can effectively improve the drug loading amount of PSO / PTX and is beneficial to magnetically target tumor sites through SPIONs@OA; secondly, coating PTX, PSO, and SPIONs@OA with PLGA can improve the stability of PSO / PTX.

[0060] (3) Add the organic phase to the aqueous phase, and after ultrasonic emulsification, evaporate to remove the organic solvent, and filter to obtain PSO / PTX targeted composite nanoparticles.

[0061] In step (3), phospholipids and modified phospholipids are used to coat the PLGA encapsulating PTX, PSO, and SPIONs@OA obtained in step (2) to form an oil-in-water emulsion layer, so as to improve the encapsulation efficiency of PTX and PSO, increase the drug loading amount, and enhance the stability of PSO and PTX. Meanwhile, the surface of modified SPIONs is easy to be modified and functionalized, and phospholipids and modified phospholipids can modify the surface of SPIONs to improve the delivery efficiency of PSO and PTX.

[0062] In summary, the PSO / PTX targeted composite nanoparticles prepared in this example firstly propose co-loading psoralen (PSO) and paclitaxel (PTX) into superparamagnetic polymer lipid nanoparticles to solve the problem of dual-drug synergistic delivery. The PSO / PTX targeted composite nanoparticles have the functions of magnetic targeting, long circulation, and slow release, and have high clinical transformation potential, indicating that the PSO / PTX targeted composite nanoparticles have good application prospects in the preparation of drugs for treating breast cancer.

[0063] As an implementation manner, the preparation method of the SPIONs@OA includes:

[0064] (2.1) Under an inert atmosphere, take FeCl2·4H2O and FeCl3·6H2O, add ultrapure water, and mix evenly to obtain solution A;

[0065] (2.2) Add ultrapure water to ammonia water and mix evenly to obtain solution B;

[0066] (2.3) Take sodium oleate, add ultrapure water, and obtain solution C after ultrasonic dissolution;

[0067] (2.4) Under an ice-water bath and stirring, add solution B to solution A to obtain a mixed solution; transfer the mixed solution to a water bath and stir to prepare a SPIONs suspension;

[0068] (2.5) Use a magnet to separate the SPIONs suspension to obtain magnetic nanoparticles and supernatant;

[0069] (2.6) Add solution C to the magnetic nanoparticles and stir and react under a water bath to obtain SPIONs@OA.

[0070] In step (2.1), the inert atmosphere is nitrogen; for example, in a reaction vessel which is a three-necked flask, place FeCl2·4H2O and FeCl3·6H2O in the three-necked flask, displace the air in the system with nitrogen, add deoxygenated ultrapure water (previously deoxygenated by nitrogen), mix evenly to obtain solution A, and place it in a 5°C ice-water bath.

[0071] In step (2.2), after obtaining solution B, it can be placed in a 5°C ice-water bath.

[0072] In step (2.4), the main processes are reaction and crystallization. Specifically, after obtaining the mixed solution, the mixed solution is transferred to a water bath, stirred at 40 °C for 0.5 h, and then stirred at 60 °C for 1 h to prepare an SPIONs suspension.

[0073] In step (2.5), the washing and separation of the SPIONs suspension are involved. Specifically, a magnet is used to separate the magnetic nanoparticles and the reaction liquid. After the nanoparticles are adsorbed, the supernatant is aspirated using a syringe. Subsequently, 20 - 50 mL of deoxygenated ultrapure water is added, and after stirring for 5 - 10 min, the magnetic nanoparticles and the supernatant are separated in the same manner. This washing process is repeated 2 - 3 times until the washed supernatant is neutral, obtaining the magnetic nanoparticles and the supernatant.

[0074] In step (2.6), the encapsulation of the magnetic nanoparticles with sodium oleate is achieved by adding solution C. Specifically, after the washing is completed, solution C is added to the magnetic nanoparticles, stirred at 50 °C for 0.5 h, separated using a magnet, and washed 2 - 5 times with ultrapure water to obtain SPIONs@OA.

[0075] As an implementation method, at least one of the following conditions is satisfied for the raw materials used in the preparation method of this example:

[0076] The modified phospholipid is DSPE-PEG2000; the second organic solvent is at least one of chloroform, dichloromethane, or acetone; the emulsifier is Tween.

[0077] Preferably, the second organic solvent is chloroform.

[0078] During the preparation of P / P-TCNs, in this example, a single-factor investigation of the formulation is simultaneously carried out, and it is found that the phospholipid dosage, PLGA dosage, and PSO / PTX dosage have a greater impact on the particle size and encapsulation efficiency of the prepared P / P-TCNs. Therefore, the phospholipid dosage, PLGA dosage, and PSO / PTX dosage are selected as the three factors for the experimental design investigation in this example.

[0079] As a preferred implementation method, the mass ratio of the phospholipid composition to PLGA is 1:1. The total mass ratio of PTX and PSO to PLGA is 0.1:1.

[0080] Within this dosage ratio range, the particle size of P / P-TCNs is more uniform, and the encapsulation efficiency is higher.

[0081] As an implementation method, adding the organic phase to the aqueous phase, followed by ultrasonic emulsification, evaporation of the organic solvent, and filtration to obtain the PSO / PTX targeted composite nanoparticles, includes:

[0082] The organic phase was added to the aqueous phase using a syringe, ultrasonic emulsification was carried out at an ultrasonic power of 225 W for 5 min, the organic solvent was removed by evaporation at 45 °C for 5 min, and PSO / PTX targeted composite nanoparticles were obtained after filtration.

[0083] Similarly, in this example, the effect of ultrasonic power on the particle size of P / P-TCNs was investigated respectively; among them, there is a certain relationship between ultrasonic power and the particle size of P / P-TCNs, and the particle size and encapsulation efficiency are the best at 225 W, so the ultrasonic power was determined to be 225 W. At the same time, when the ultrasonic time exceeds 5 min, the particle size of P / P-TCNs increases with the increase of ultrasonic time and the encapsulation efficiency decreases, so the ultrasonic time was selected to be 5 min.

[0084] Generally, in the above steps (1)-(4), stirring will be carried out during the reaction process to accelerate the reaction or make the reaction more complete.

[0085] Secondly, the PSO / PTX targeted composite nanoparticles (P / P-TCNs) in the second aspect of this example will be described.

[0086] PSO / PTX Targeted Composite Nanoparticles

[0087] As described in the first aspect, in this example, modified SPIONs (SPIONs@OA) were used to improve the delivery efficiency of PSO / PTX. Specifically, PTX, PSO and SPIONs@OA were coated by PLGA, and then PLGA was modified by phospholipids and modified phospholipids (forming an oil-in-water emulsion layer) to improve the encapsulation efficiency of PTX and PSO, so as to improve the drug loading capacity and the stability of PSO and PTX. At the same time, the surface of modified SPIONs is easy to be modified and functionalized, and phospholipids and modified phospholipids can modify the surface of SPIONs to improve the delivery efficiency of PSO and PTX.

[0088] Therefore, the PSO / PTX targeted composite nanoparticles prepared in this example first proposed to co-load psoralen (PSO) and paclitaxel (PTX) in superparamagnetic polymer lipid nanoparticles, solving the problem of dual-drug synergistic delivery. The PSO / PTX targeted composite nanoparticles have both magnetic targeting, long circulation and sustained release functions, and have high clinical transformation potential, indicating that the PSO / PTX targeted composite nanoparticles have good application prospects in the preparation of drugs for treating breast cancer.

[0089] Next, the use of the PSO / PTX targeted composite nanoparticles in the third aspect of this example will be described.

[0090] Use

[0091] As described in the second aspect, the PSO / PTX targeted composite nanoparticles prepared in this example have multiple advantages. For example: (1) It can improve the drug loading and stability of PSO / PTX, magnetically target the tumor site, increase the accumulation of the drug (PSO / PTX) at the tumor site, and reduce the damage to normal tissues; (2) The chemical toxicity of PSO / PTX and the magnetic induction heat of SPIONs have a synergistic therapeutic effect, improving the efficacy of the drug and reducing the toxic and side effects.

[0092] Based on this, the PSO / PTX targeted composite nanoparticles provided in this example can be used to prepare drugs for preventing, alleviating or treating breast cancer.

[0093] It should be particularly noted that the PSO / PTX targeted composite nanoparticles of this example can be made into drugs for inhibiting the growth of MDA-MB-231 cells.

[0094] Exemplarily, the above-mentioned drug is an injectable solution or a drug for oral administration.

[0095] Generally, when the above-mentioned PSO / PTX targeted composite nanoparticles are made into corresponding drugs, the drugs also include pharmaceutically acceptable carriers and / or adjuvants.

[0096] For the drugs described in this example, the drugs take the PSO / PTX targeted composite nanoparticles as the active ingredient, excluding changes in the formulation system and administration methods, medicinal salts after simple chemical modification and adjustment of the above-mentioned PSO / PTX targeted composite nanoparticles, and the combination of multiple compounds, etc.

[0097] For example, in this example, one or more compounds in the PSO / PTX targeted composite nanoparticles of this example can be formulated as active ingredients in non-toxic, inert and pharmaceutically acceptable carriers and / or adjuvants; the prepared drugs can be administered through conventional routes, including but not limited to oral, intramuscular, intraperitoneal, intravenous, subcutaneous, intradermal or topical administration.

[0098] For example, when the dosage form of the drug in this example is a drug for oral administration, it contains a safe and effective amount of PSO / PTX targeted composite nanoparticles and pharmaceutically acceptable carriers and / or adjuvants. The drugs for oral administration can be made into common dosage forms such as tablets, pills, powders, granules, capsules, emulsions, syrups, ointments, suppositories, etc.; in this example, no specific limitations are placed on the carriers and / or adjuvants, and the carriers and / or adjuvants can be adaptively adjusted according to the specific drug dosage form.

[0099] Generally, an "effective amount" of a compound (PSO / PTX targeted composite nanoparticles) refers to an amount sufficient to elicit a desired biological response. As understood by those of ordinary skill in the art, the effective amount of the compound in this example can vary depending on the following factors: for example, components such as the vehicle in the drug, as well as the age, health status of the subject, and breast cancer symptoms.

[0100] Among them, the effective amount includes a therapeutically effective amount and a prophylactically effective amount.

[0101] Unless otherwise specified, the "therapeutically effective amount" of the compound used in this example is an amount sufficient to provide a benefit during the treatment of breast cancer, or the amount that minimizes the improvement or remission of one or more symptoms (manifestations) related to the breast cancer state. The "prophylactically effective amount" of the compound used in this example is an amount sufficient to prevent the occurrence of breast cancer, or an amount sufficient to prevent one or more symptoms related to the occurrence of breast cancer state.

[0102] It can be understood that the drug of this example can also be made into an injection. For example, PSO / PTX targeted composite nanoparticles can be made into corresponding injections with water for injection, normal saline, and glucose water under a sterile operating environment. The above injections can be prepared by conventional methods.

[0103] The following will further elaborate on this application in combination with examples. It should be understood that these examples are only used to illustrate this application and not to limit the scope of this application.

[0104] In the following examples, the materials, reagents, and instruments used, unless otherwise specified, can be obtained from commercial sources.

[0105] In the following examples, unless otherwise specified, the modified phospholipid is DSPE-PEG2000, and the addition rate of the liquid is 5 mL / min.

[0106] In the following examples, the ultrasonic instrument used is the SONICS VCX750 ultrasonic cell disruptor from the United States, and its rated power is 750 W; the ultrasonic cell disruptor represents the power size as a percentage, 10% - 100%, 100% represents 550 W, and so on, 20% represents 150 W, 30% represents 225 W, 40% represents 80 W.

[0107] In the following examples, the test method for the encapsulation efficiency was as follows: Absorb an appropriate amount of the P / P-TCNs solution into a 30KD ultrafiltration centrifuge tube, centrifuge at 12000 r / min for 20 min, take 20 μL of the filtrate and inject it into the chromatograph to determine the amount of free drug of PSO / PTX (Wf); additionally, accurately measure 1.0 mL of the P / P-TCNs solution into a 10 mL volumetric flask, add the mobile phase (acetonitrile: water (55:45)) to make up the volume, ultrasonically demulsify, filter through a 0.45 μm filter membrane, take 20 μL of the filtrate and inject it into the chromatograph to calculate the total amount of drug of PSO / PTX (Wt).

[0108] Among them, the test conditions of the chromatograph were as follows: chromatographic column: Phenomenex C18 chromatographic column (250×4.6 mm, 4 μm), mobile phase: acetonitrile: water (55:45), detection wavelength: 227 nm, flow rate: 1.0 mL / min, injection volume: 20 μL, column temperature: 25 °C.

[0109] And the drug encapsulation efficiency was calculated according to the following formula:

[0110] The present application will be further described below in conjunction with specific examples.

[0111] Example 1: Preparation of SPIONs@OA.

[0112] The preparation method of SPIONs@OA specifically included the following steps:

[0113] 1.1 Preparation of solution A: Weigh 0.696 g of FeCl2·4H2O and 1.351 g of FeCl3·6H2O into a three-necked flask, displace the air in the system with nitrogen, add the prescribed amount of 50 mL of deoxygenated ultrapure water (deoxygenated by nitrogen in advance), and place it in an ice bath at 5 °C.

[0114] 1.2 Preparation of solution B: Add 14 mL of ammonia water to 50 mL of deoxygenated ultrapure water and place it in an ice bath at 5 °C.

[0115] 1.3 Preparation of solution C: Weigh 1.5 g of sodium oleate and add it to 50 mL of ultrapure water, and dissolve it by ultrasonic treatment.

[0116] 1.4 Mixing: Under the condition of an ice bath at 5 °C and stirring, add solution B to solution A (control the addition rate to be 5 mL / min), and adjust the pH value of the solution to 11.

[0117] 1.5 Reaction and crystallization: Transfer the solution obtained in step 1.4 to a water bath, stir at 40 °C for 0.5 h, and then stir at 60 °C for 1 h to prepare a SPIONs suspension.

[0118] 1.6. Washing: Use a magnet to separate the magnetic nanoparticles from the reaction liquid. After the magnetic nanoparticles are adsorbed, use a syringe to extract the supernatant. Subsequently, add 20 - 50 mL of deoxygenated ultrapure water, stir for 10 min, and then separate the magnetic nanoparticles and the supernatant in the same way. Wash repeatedly 2 - 3 times until the washed supernatant is neutral.

[0119] 1.7. Sodium oleate coating: After washing, add solution C to the above-mentioned magnetic nanoparticles, stir in a water bath at 50 °C for 0.5 h, separate using a magnet, and wash 2 - 3 times with ultrapure water, and then obtain SPIONs@OA by centrifugal separation.

[0120] The SPIONs@OA prepared in this example is used for the subsequent preparation of related nanoparticles.

[0121] Example 2: Preparation of P / P-TCNs (PSO / PTX targeted composite nanoparticles).

[0122] In this example, a single-factor investigation was carried out on the preparation method of P / P-TCNs.

[0123] In the process of preparing P / P-TCNs in this example, a single-factor investigation of its prescription was carried out. Combining with the Box-Behnken experimental factor level table in Table 1, it was found that the amounts of phospholipids, PLGA, and PSO / PTX had a greater impact on the particle size and encapsulation efficiency of the prepared P / P-TCNs. Therefore, the amounts of phospholipids, PLGA, and PSO / PTX were selected as the three factors investigated in this experimental design, and a response surface Box-Behnken experiment was designed for process and prescription optimization.

[0124] Table 1: Box-Behnken experimental factor level table

[0125]

[0126] Based on the above investigation results of the Box-Behnken experimental factor levels, this example further set up Examples 2.1 - 2.7 for single-factor investigation.

[0127] Example 2.1: Verify the influence of the amount of phospholipids on the preparation of P / P-TCNs.

[0128] Weigh different prescription amounts of phospholipids (40 mg, 80 mg, 120 mg) and DSPE-PEG2000 (10 mg, 20 mg, 30 mg). The total amount of phospholipids used is (50 mg, 100 mg, 150 mg). Add them to 0.3 mL of absolute ethanol for dissolution. After complete dissolution, add 50 mL of purified water. Stir and mix well, then add 0.2 mL of Tween and stir evenly. Keep it at 2 - 8 °C to obtain the aqueous phase. Weigh 100 mg of PLGA and dissolve it in 1.5 mL of chloroform by ultrasonic dissolution. After dissolution, add 5 mg of PTX and 5 mg of PSO and dissolve them ultrasonically. After dissolution, add 0.1 g of SPIONs@OA (prepared in Example 1) and disperse it ultrasonically to obtain the organic phase. Under the ultrasonic probe, slowly inject the organic phase into the aqueous phase using a syringe. The ultrasonic power is 30% (225 W) and the ultrasonic time is 5 min. After ultrasonic treatment, rotary evaporate for 5 min at 45 °C to remove the organic solvent. Sterilize and filter using a 0.22 μm polyethersulfone filter membrane to obtain P / P-TCNs.

[0129] After preparing P / P-TCNs, use a laser particle size analyzer (ZS90, Malvern, UK) to measure the particle size, PDI, and zeta potential of P / P-TCNs, and test the encapsulation efficiency of PTX and PSO. The test results are shown in Table 2.

[0130] Table 2: Particle size, zeta potential, and encapsulation efficiency of P / P-TCNs prepared in Example 2.1

[0131]

[0132] According to the test results in Table 2, the amount of phospholipids used has a significant impact on the preparation of P / P-TCNs. Among them, too little phospholipid may not be able to completely encapsulate PLGA nanoparticles, and too much phospholipid may form liposomes. Experiments show that when the amount of phospholipids and DSPE-PEG2000 used is 100 mg, the particle size is more uniform, which is 188.7 ± 2.13 nm; the particle size distribution is relatively uniform, with a PDI of 0.161 ± 0.03; the zeta potential is -40.2 ± 0.88 mA, the encapsulation efficiency of PSO is 82.1%, and the encapsulation efficiency of PTX is 84.3%.

[0133] Example 2.2: Verify the influence of the amount of PLGA used on the preparation of P / P-TCNs.

[0134] Weigh 80 mg of phospholipids and 20 mg of DSPE-PEG2000, add them to 0.3 mL of absolute ethanol for dissolution. After complete dissolution, add 50 mL of purified water, stir and mix well, then add 0.2 mL of Tween, stir evenly, and keep it at 2 - 8 °C to obtain the aqueous phase. Weigh different amounts of PLGA (50 mg, 100 mg, 150 mg) according to the prescription and dissolve them in 1.5 mL of chloroform, and dissolve them by ultrasonic treatment. After dissolution, add 5 mg of PTX and 5 mg of PSO, and dissolve them by ultrasonic treatment. After dissolution, add 0.1 g of SPIONs@OA and disperse it by ultrasonic treatment to obtain the organic phase. Under the ultrasonic probe, slowly inject the organic phase into the aqueous phase using a syringe, with an ultrasonic power of 30% and an ultrasonic time of 5 min. After ultrasonic treatment, rotate and evaporate for 5 min at 45 °C to remove the organic solvent. Sterilize and filter using a 0.22 μm polyethersulfone filter membrane to obtain P / P-TCNs.

[0135] After preparing P / P-TCNs, use a laser particle size analyzer (ZS90, Malvern, UK) to measure the particle size, PDI, and zeta potential of P / P-TCNs, and test the encapsulation efficiency of PTX and PSO. The test results are shown in Table 3.

[0136] Table 3: Particle size, zeta potential, and encapsulation efficiency of P / P-TCNs prepared in Example 2.2

[0137]

[0138] In this example, the effects of 50 mg, 100 mg, and 150 mg of PLGA dosage on the particle size of P / P-TCNs were investigated respectively. As shown in the test results of Table 3, the above results indicate that there is a direct relationship between the PLGA dosage and the particle size and particle size distribution of P / P-TCNs. Among them, when the dosage is 100 mg, the particle size, particle size distribution, and encapsulation efficiency perform the best.

[0139] Example 2.3: Verify the influence of the dosage of PSO / PTX on the preparation of P / P-TCNs.

[0140] Weigh 80 mg of phospholipids and 20 mg of DSPE-PEG2000, add them to 0.3 mL of absolute ethanol for dissolution. After complete dissolution, add 50 mL of purified water, stir and mix well, then add 0.2 mL of Tween, stir evenly, and keep it at 2 - 8 °C to obtain the aqueous phase. Weigh 100 mg of PLGA and dissolve it in 1.5 mL of chloroform by ultrasonic dissolution. After dissolution, add different prescription amounts of PTX (2.5 mg, 5 mg, 7.5 mg) and PSO (2.5 mg, 5 mg, 7.5 mg), and dissolve them by ultrasonic. After dissolution, add 0.1 g of SPIONs@OA and disperse it by ultrasonic to obtain the organic phase. Under the ultrasonic probe, slowly inject the organic phase into the aqueous phase using a syringe, with an ultrasonic power of 30% and an ultrasonic time of 5 min. After ultrasonic treatment, rotary evaporate for 5 min at 45 °C to remove the organic solvent. Sterilize and filter using a 0.22 μm polyethersulfone filter membrane to obtain P / P-TCNs.

[0141] After preparing P / P-TCNs, use a laser particle size analyzer (ZS90, Malvern, UK) to measure the particle size, PDI, and zeta potential of P / P-TCNs, and test the encapsulation efficiency of PTX and PSO. The test results are shown in Table 4.

[0142] Table 4: Particle size, zeta potential, and encapsulation efficiency of P / P-TCNs prepared in Example 2.3

[0143]

[0144] In this example, the effects of the dosages of PTX and PSO on the particle size of P / P-TCNs were investigated respectively. Among them, in Table 4 above, the dosage of PSO / PTX being 5 mg means PTX 2.5 mg + PSO 2.5 mg, the dosage of PSO / PTX being 10 mg means PTX 5 mg + PSO 5 mg, and the dosage of PSO / PTX being 15 mg means PTX 7.5 mg + PSO 7.5 mg.

[0145] Combined with the test results shown in Table 4, it shows that there is a direct relationship between the dosages of PTX and PSO and the particle size of P / P-TCNs, and when the dosages of PTX and PSO are 10 mg, the particle size is the smallest and the encapsulation efficiency is the highest.

[0146] Example 2.4: Verify the effect of the emulsifier dosage on the preparation of P / P-TCNs.

[0147] Weigh 80 mg of phospholipids and 20 mg of DSPE-PEG2000, add them to 0.3 mL of absolute ethanol for dissolution. After complete dissolution, add 50 mL of purified water. After stirring and mixing evenly, add different amounts of Tween (0.1 mL, 0.2 mL, 0.3 mL), stir evenly, and keep it at 2 - 8 °C to obtain the aqueous phase. Weigh 100 mg of PLGA and dissolve it in 1.5 mL of chloroform by ultrasonic dissolution. After dissolution, add 5 mg of PTX and 5 mg of PSO, and dissolve them by ultrasonic treatment. After dissolution, add 0.1 g of SPIONs@OA and disperse it by ultrasonic treatment to obtain the organic phase. Under the ultrasonic probe, slowly inject the organic phase into the aqueous phase using a syringe, with an ultrasonic power of 30% and an ultrasonic time of 5 min. After ultrasonic treatment, evaporate the organic solvent by rotary evaporation at 45 °C for 5 min. Sterilize and filter using a 0.22 μm polyethersulfone filter membrane to obtain P / P-TCNs.

[0148] After preparing P / P-TCNs, use a laser particle size analyzer (ZS90, Malvern, UK) to measure the particle size, PDI, and zeta potential of P / P-TCNs, and test the encapsulation efficiency of PTX and PSO. The test results are shown in Table 5.

[0149] Table 5: Particle size, zeta potential, and encapsulation efficiency of P / P-TCNs prepared in Example 2.4

[0150]

[0151] In this example, the effects of the amounts of emulsifier of 0.1 mL, 0.2 mL, and 0.3 mL on the particle size of P / P-TCNs were investigated respectively. As shown in the test results of Table 5, it shows that the relationship between the amount of emulsifier and the particle size of P / P-TCNs is not obvious.

[0152] Example 2.5: Verify the effect of ultrasonic power on the preparation of P / P-TCNs.

[0153] Weigh 80 mg of phospholipids and 20 mg of DSPE-PEG2000, add them to 0.3 mL of absolute ethanol for dissolution. After complete dissolution, add 50 mL of purified water. After stirring and mixing evenly, add 0.2 mL of Tween, stir evenly, and keep it at 2 - 8 °C to obtain the aqueous phase. Weigh 100 mg of PLGA and dissolve it in 1.5 mL of chloroform by ultrasonic dissolution. After dissolution, add 5 mg of PTX and 5 mg of PSO, and dissolve them by ultrasonic treatment. After dissolution, add 0.1 g of SPIONs@OA and disperse it by ultrasonic treatment to obtain the organic phase. Under the ultrasonic probe, slowly inject the organic phase into the aqueous phase using a syringe, and carry out ultrasonic treatment for 5 min at different ultrasonic powers (20%, 30%, 40%). After ultrasonic treatment, evaporate the organic solvent by rotary evaporation at 45 °C for 5 min. Sterilize and filter using a 0.22 μm polyethersulfone filter membrane to obtain P / P-TCNs.

[0154] After preparing P / P-TCNs, a laser particle size analyzer (ZS90, Malvern, UK) was used to measure the particle size, PDI and potential of P / P-TCNs, and the encapsulation efficiency of PTX and PSO was tested. The test results are shown in Table 6.

[0155] Table 6: Particle size, potential, and encapsulation efficiency of P / P-TCNs prepared in Example 2.5

[0156]

[0157] In this example, the effects of ultrasonic powers of 30%, 40% and 50% on the particle size of P / P-TCNs were investigated respectively. As shown in the test results of Table 6, it shows that there is a certain relationship between the ultrasonic power and the particle size of P / P-TCNs, and the particle size and encapsulation efficiency are the best at 30%. Therefore, the ultrasonic power was determined to be 30%.

[0158] Example 2.6: Verify the effect of ultrasonic time on the preparation of P / P-TCNs.

[0159] Weigh 80 mg of phospholipids and 20 mg of DSPE-PEG2000, add them to 0.3 mL of absolute ethanol and dissolve. After complete dissolution, add 50 mL of purified water, stir and mix well, then add 0.2 mL of Tween, stir evenly, and keep it at 2 - 8 °C to obtain the aqueous phase. Weigh 100 mg of PLGA and dissolve it in 1.5 mL of chloroform by ultrasonic dissolution. After dissolution, add 5 mg of PTX and 5 mg of PSO and dissolve them by ultrasonic. After dissolution, add 0.1 g of SPIONs@OA and disperse it by ultrasonic to obtain the organic phase. Under the ultrasonic probe, the organic phase was slowly injected into the aqueous phase using a syringe, and ultrasonicated for different times (3 min, 5 min, 10 min) under the condition of ultrasonic power of 30%. After ultrasonication, the organic solvent was removed by rotary evaporation at 45 °C for 5 min. Sterile filtration was carried out using a 0.22 μm polyethersulfone membrane to obtain P / P-TCNs.

[0160] After preparing P / P-TCNs, a laser particle size analyzer (ZS90, Malvern, UK) was used to measure the particle size, PDI and potential of P / P-TCNs, and the encapsulation efficiency of PTX and PSO was tested. The test results are shown in Table 7.

[0161] Table 7: Particle size, potential, and encapsulation efficiency of P / P-TCNs prepared in Example 2.6

[0162]

[0163] In this example, while fixing other conditions, the ultrasonic treatment times were set to 3, 5, and 10 min respectively to prepare samples, and the average particle size and encapsulation efficiency of P / P-TCNs prepared under different conditions were measured. The results are shown in Table 7. When the ultrasonic treatment time exceeds 5 min, the particle size of P / P-TCNs increases with the increase of ultrasonic treatment time and the encapsulation efficiency decreases. Therefore, the ultrasonic treatment time of 5 min was selected.

[0164] Example 3: Optimization by Box-Behnken response surface experiment.

[0165] In this example, the preparation method of P / P-TCNs in Example 2 was optimized using Box-Behnken response surface, and the measured particle size and encapsulation efficiency are shown in Table 8.

[0166] Table 8: Design results of Box-Behnken response surface

[0167]

[0168] This example involves the establishment of a regression equation, specifically:

[0169] Linear regression was performed with independent variables (phospholipid dosage, PLGA dosage, PSO / PTX dosage) and dependent variables (particle size Y1, PSO encapsulation efficiency Y2, PTX encapsulation efficiency Y3) to evaluate the fitting effect of the model. Among them, the results of the quadratic multiple regression equation are as follows:

[0170] Y1 = 188.60 + 33.92A + 6.62B + 1.17C - 8.85AB - 1.70AC + 2.60BC + 36.10A 1 0.05B 2 +

[0171] 26.40C 2 , R 2 = 0.9982, where A, B, AB, A 2 , B 2 , C 2 are significant (p < 0.05).

[0172] Y2 = 82.17 - 3.91A - 1.11B - 0.5750C - 0.9000AB - 1.47AC + 1.43BC - 8.93A 2 - 0.8333B 2

[0173] - 9.21C 2 , R 2 = 0.9993, where A, B, C, AB, AC, BC, A 2 , B 2 , C2 All were significant (p < 0.05).

[0174] Y3 = 85.40 - 3.69A - 0.9375B - 0.7000C - 2.6AB - 1.27AC + 1.72BC - 10.02A 1 .22B 2 -9.6

[0175] 5C 2 , R 2 = 0.9964, where A, B, AB, AC, BC, A 2 , B 2 , C 2 were significant (p < 0.05).

[0176] In this example, analysis was carried out through Design-Expert 13 software in combination with the fitting results, and the three-dimensional response surface diagram of particle size and encapsulation efficiency is as follows Figures 1 - 3 .

[0177] Among them, Figure 1 , Figure 2 and Figure 3 in A is the three-dimensional surface diagram of the dosage of PLGA and phospholipid, Figure 1 , Figure 2 and Figure 3 in B is the three-dimensional surface diagram of the dosage of PSO / PTX and phospholipid, Figure 1 , Figure 2 and Figure 3 in C is the three-dimensional surface diagram of the dosage of PSO / PTX and PLGA. In Figure 1 , Figure 2 and Figure 3 the surfaces of A, B, C, D, E, and F are all relatively curved, indicating that the dosages of phospholipid, PSO / PTX, and PLGA have a greater impact on the particle size, encapsulation efficiency of PSO and PTX.

[0178] Specifically, Figure 1 shows the response surface diagram of the particle size of P / P-TCNs; Figure 1 in A, B, and C are three-dimensional response surface diagrams. D, E, and F are contour diagrams. A and D reflect the relationship between the dosage of PLGA and phospholipid, B and E reflect the relationship between the dosage of PSO / PTX and phospholipid, and C and F reflect the relationship between the dosage of PSO / PTX and PLGA.

[0179] Figure 2 shows the response surface diagram of the PSO encapsulation efficiency of P / P-TCNs; Figure 2Among them, A, B, and C are three-dimensional response surface diagrams. D, E, and F are contour diagrams. A and D reflect the relationship between the dosage of PLGA and phospholipids, B and E reflect the relationship between the dosage of PSO / PTX and phospholipids, and C and F reflect the relationship between the dosage of PSO / PTX and PLGA.

[0180] Figure 3 Shows the response surface diagram of the PTX encapsulation efficiency of P / P-TCNs; Figure 3 Among them, A, B, and C are three-dimensional response surface diagrams. D, E, and F are contour diagrams. A and D reflect the relationship between the dosage of PLGA and phospholipids, B and E reflect the relationship between the dosage of PSO / PTX and phospholipids, and C and F reflect the relationship between the dosage of PSO / PTX and PLGA.

[0181] Example 4: Based on the above response surface design results, the preparation process of P / P-TCNs was verified.

[0182] According to the experimental results of Example 2 and Example 3, the system predicted the optimal formulation: lipid dosage: 101.5 mg (81.2 mg of phospholipid and 20.3 mg of DSPE-PEG2000), PLGA dosage: 66.8 mg; PSO / PTX dosage: 9.6 mg (4.8 mg each of PSO and PTX). The system predicted that the encapsulation efficiency of PSO was 82.5% and that of PTX was 85.5%, with an accuracy of 99.7%.

[0183] According to the optimal formulation predicted by the system, three batches of samples were prepared and evaluated according to the preparation method of Example 2 under the optimal conditions of Example 2 (ultrasonic power 30% and ultrasonic time 5 min). The results are shown in Table 9. As can be seen from Table 9, all deviations are less than ±5%, indicating that the model can better reflect the relationship between the indicators and factors.

[0184] Table 9: Comparison of predicted values and true values (n = 3)

[0185] Index Predicted Value True Value Deviation (%) PSO Encapsulation Efficiency (%) 82.5 81.3±1.36 -1.5 PTX Encapsulation Efficiency (%) 85.5 84.7±0.94 -0.9

[0186] Example 5: Stability test.

[0187] The P / P-TCNs prepared in Example 4 were stored at 4 °C and room temperature, and the changes in particle size and encapsulation efficiency were tested after the 1st - 3rd week. Among them, as shown in Table 10, there were no significant changes in particle size and encapsulation efficiency within 3 weeks. Therefore, the prepared P / P-TCNs are relatively stable when stored at 4 °C and room temperature.

[0188] Table 10: Stability of P / P-TCNs (n = 3)

[0189]

[0190] Example 6: Infrared test.

[0191] In this example, a Fourier transform infrared spectrometer (Nicoleti S50, Thermo Fisher Scientific, USA) was used to analyze PSO, PTX, PSO + PTX, and P / P-TCNs prepared in Example 2. The test results are as Figure 4 shown.

[0192] Figure 4 The figure shows a schematic diagram of the analysis test results of the infrared spectrum. Figure 4 In the figure, A is PSO, B is PTX, C is PSO + PTX, and L is P / P-TCNs.

[0193] Figure 4 In the figure, curve A has a strong absorption peak at 1720 cm -1 , which is the C=O characteristic peak of PSO. The 1720 and 1650 cm -1 in (B) are the characteristic absorption peaks of the ester group C=O and amide group C=O of paclitaxel, respectively. (C) is a physical mixture of PSO and PTX. Therefore, the characteristic absorption peaks at 1720 and 1650 cm -1 both appear, while the characteristic absorption peaks at 1720 and 1650 cm -1 are not seen in (L), indicating that PSO and PTX have been well encapsulated in the P / P-TCNs nanoparticles.

[0194] Example 7: Cytotoxicity test.

[0195] The in vitro cytotoxicity test in this example was performed using the CCK-8 method. Triple-negative breast cancer MDA-MB-231 cells were seeded in 96-well plates (5.0×10 3 cells / well), cultured for 24 h to allow them to adhere, and then different concentrations of P / P-TCNs were administered (for specific concentrations, please refer to Figure 5 ). Blank medium and blank cells were set as controls. After culturing each group in an incubator for 48 h, CCK-8 solution (10 μL) was added to each sample well, and the 96-well plate was returned to the incubator for continued culture for 4 hours. A full-wavelength microplate reader (ReadMax 1200, Shanghai Flash Spectrum Biotechnology Co., Ltd., China) was used to record the absorbance (OD) of each sample at 450 nm. The cytotoxicity was calculated as follows:

[0196] Cell survival rate (%) = (ODa - ODb) / (ODc - ODb) × 100;

[0197] Among them, ODa is the OD value of the sample to be measured (containing cells, CCK-8, and DMEM medium with drugs), ODc is the OD value of the control group (containing DMEM medium and CCK-8 solution), and ODb is the OD value of the blank well (containing cells, DMEM medium, and CCK-8 solution).

[0198] The test results are as Figure 5 shown.

[0199] According to Figure 5 the experimental results, in this example, a CCK-8 kit was used to detect the cytotoxicity of P / P-TCNs on MDA-MB-231 cells. The survival rate of MDA-MB-231 cells decreased with the increase in the concentration of P / P-TCNs. That is, the higher the concentration of P / P-TCNs, the more obvious the inhibitory effect on MDA-MB-231 cells. Combining Figure 5 the test results, the IC50 of P / P-TCNs on MDA-MB-231 cells was calculated to be 13.24 ug / mL. Therefore, P / P-TCNs administration has good efficacy against the growth of MDA-MB-231 cells.

[0200] In summary, in this example, PLGA coated with PTX, PSO, and SPIONs@OA was modified by phospholipids and modified phospholipids to improve the encapsulation efficiency of PTX and PSO, thereby increasing the drug loading capacity and the stability of PSO and PTX. At the same time, the surface of modified SPIONs is easy to be modified and functionalized, and phospholipids and modified phospholipids can modify the surface of SPIONs to improve the delivery efficiency of PSO and PTX.

[0201] In summary, the PSO / PTX targeted composite nanoparticles prepared in this example firstly proposed co-loading psoralen (PSO) and paclitaxel (PTX) in superparamagnetic polymer lipid nanoparticles, solving the problem of dual-drug synergistic delivery. The PSO / PTX targeted composite nanoparticles have both magnetic targeting, long circulation, and sustained release functions, and have high clinical transformation potential, indicating that the PSO / PTX targeted composite nanoparticles have good application prospects in the preparation of drugs for treating breast cancer.

[0202] The technical solutions provided by the embodiments of the present application have been introduced in detail above. Specific examples are used in this article to elaborate on the principles and implementation manners of the embodiments of the present application. The descriptions of the above embodiments are only applicable to help understand the principles of the embodiments of the present application; at the same time, for those of ordinary skill in the art, according to the embodiments of the present application, there will be changes in the specific implementation manners and application scopes. In summary, the content of this specification should not be construed as a limitation to the present application.

Claims

1. A preparation method of PSO / PTX targeted composite nanoparticles, characterized in that, The preparation method includes: Dissolve the phospholipid composition in a first organic solvent. After dissolution, add water and an emulsifier, and mix evenly to obtain an aqueous phase; wherein, the phospholipid composition includes phospholipids and modified phospholipids; Dissolve PLGA in a second organic solvent. After dissolution, add PTX, PSO, and SPIONs@OA, and mix evenly to obtain an organic phase; Add the organic phase to the aqueous phase, and obtain PSO / PTX targeted composite nanoparticles after ultrasonic emulsification, evaporation of the organic solvent, and filtration.

2. The preparation method according to claim 1, wherein The preparation method of the SPIONs@OA includes: Under an inert atmosphere, take FeCl2·4H2O and FeCl3·6H2O, add ultrapure water, and mix evenly to obtain solution A; Add ultrapure water to ammonia water, and mix evenly to obtain solution B; Take sodium oleate and add ultrapure water, and obtain solution C after ultrasonic dissolution; Under an ice-water bath and stirring, add solution B to solution A to obtain a mixed solution; transfer the mixed solution to a water bath and stir to prepare a SPIONs suspension; Use a magnet to separate the SPIONs suspension to obtain magnetic nanoparticles and supernatant; Add solution C to the magnetic nanoparticles, and stir and react under a water bath to obtain SPIONs@OA.

3. The preparation method according to claim 1, characterized in that, The modified phospholipid is DSPE-PEG2000.

4. The preparation method according to claim 1 or 3, characterized in that, The mass ratio of the phospholipid composition to PLGA is 1:

1.

5. The preparation method according to claim 1, wherein Meet at least one of the following conditions: The first organic solvent is anhydrous ethanol and / or methanol; The second organic solvent is at least one of chloroform, dichloromethane, or acetone; The emulsifier is Tween.

6. The preparation method according to claim 1, characterized in that, The total mass ratio of PTX and PSO to PLGA is 0.1:

1.

7. The preparation method according to claim 1, characterized in that, The step of adding the organic phase to the aqueous phase, and obtaining PSO / PTX targeted composite nanoparticles after ultrasonic emulsification, evaporation of the organic solvent, and filtration includes: Add the organic phase to the aqueous phase, perform ultrasonic emulsification at an ultrasonic power of 225 W for 5 min, evaporate the organic solvent at 45 °C for 5 min, and obtain PSO / PTX targeted composite nanoparticles after filtration.

8. A PSO / PTX targeted composite nanoparticle, characterized in that, The PSO / PTX targeted composite nanoparticles are prepared by the preparation method described in any one of claims 1-7.

9. Use of the PSO / PTX targeted composite nanoparticles according to claim 8 in the preparation of a drug for preventing, alleviating, or treating breast cancer.

10. The use according to claim 9, characterized in that, The drug is a drug for inhibiting the growth of MDA-MB-231 cells.

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