Preparation and anti-breast cancer application of Ag2Fe2O4-TiO2 (at) Rh2-PTX lipid nanoparticles
By preparing Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles, combined with photothermal and ROS generation, the problem of poor resistance and treatment effect of triple-negative breast cancer to existing chemotherapy drugs was solved, and efficient and targeted multimodal therapeutic effects were achieved.
Patent Information
- Application Number
- CN202411913789.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-24
- Publication Date
- 2025-03-28
AI Technical Summary
Due to the lack of targeted treatment methods for triple-negative breast cancer, existing chemotherapy drugs lack tumor specificity, which can easily cause damage to normal cells, and TNBC cells are prone to resistance to chemotherapy drugs, resulting in unsatisfactory treatment results.
Ultrasonic chemical reduction method was used to prepare the Ag2Fe2O4-TiO2 magnetic nanoparticle core, and paclitaxel and ginsen saponin Rh2 were coated on the core surface by ethanol injection to form Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles, combining photothermal effect and ROS generation to achieve multi-mode treatment.
It significantly reduces the invasion and migration ability of breast cancer cells, induces apoptosis of tumor cells through photothermal and ROS generation, improves the targeting and safety of treatment, and the tumor suppression rate reaches 82.56%, which has excellent anti-breast cancer effect and biosafety.
Smart Images
Figure CN119700673A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of medical technology, and specifically relates to the preparation of Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles and their application in fighting breast cancer. Background Art
[0002] Breast cancer is one of the most common malignant tumors in women worldwide, and its morbidity and mortality have been increasing year by year. Among them, triple-negative breast cancer (TNBC), as a special subtype of breast cancer, lacks the expression of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor-2 (HER2), which makes it impossible to intervene clinically through endocrine therapy or anti-HER2 targeted therapy. TNBC is highly heterogeneous and invasive, prone to recurrence and metastasis, and has a poor prognosis for patients. It is a major problem in the current research and treatment of breast cancer. At present, the main treatment method for TNBC is still chemotherapy, but chemotherapy drugs (such as paclitaxel (PTX)) lack tumor specificity and are prone to damage normal cells. TNBC cells are prone to drug resistance to chemotherapy drugs, resulting in unsatisfactory treatment effects. At the same time, TNBC is prone to metastasize to important organs such as the lungs, bones and liver, which further increases the difficulty of treatment. Therefore, the development of treatment technologies with strong targeting, low toxicity and side effects, and the ability to overcome drug resistance has become a research hotspot for the treatment of TNBC.
[0003] In recent years, nanotechnology has received extensive attention in tumor diagnosis and treatment. Nanocarriers can significantly improve the therapeutic effect of anticancer drugs by enhancing the solubility, stability and targeting of drugs. Among them, magnetic nanoparticles (such as ferrite-based nanomaterials) have been used in the diagnosis and treatment of tumors due to their magnetic imaging properties and drug delivery capabilities. At the same time, photothermal therapy, as an emerging tumor treatment method, can induce nanomaterials to produce local high temperatures through near-infrared light NIR, thereby selectively killing tumor cells. Therefore, if it can be combined with immunomodulatory molecules for treatment, it is expected to improve the anti-tumor efficacy. However, the biocompatibility and targeting of most magnetic nanoparticles are still poor, and their drug loading capacity and combined treatment effects have not yet been able to meet clinical needs. Therefore, it is of great significance to develop a targeted nano drug carrier with both diagnostic and therapeutic functions.
[0004] Ferrite materials have good photothermal conversion efficiency in the near-infrared region. This property stems from its unique electronic structure, which enables it to absorb NIR light and convert light energy into heat energy, thereby achieving a rapid increase in local temperature. In tumor treatment, the photothermal effect of silver ferrite can directly kill cancer cells through local heating, and can synergize with a variety of treatment methods such as chemotherapy and immunotherapy to improve the therapeutic effect. At the same time, compared with traditional surgery and radiotherapy, photothermal therapy has less trauma and strong targeting, which can reduce damage to normal tissues. Furthermore, the surface of the nanoparticles of silver ferrite materials has rich functional groups, which can be combined with drug molecules through chemical modification or targeted delivery through magnetic manipulation, and the drugs combined can be chemotherapy drugs, such as paclitaxel, or natural products with anticancer activity, such as ginsenoside Rh2. The drug delivery system constructed in this way can not only increase the enrichment concentration of drugs in the tumor site, but also reduce systemic toxicity. In addition, by combining with photothermal therapy, the permeability of the tumor microenvironment can be enhanced, further promoting the release and absorption of drugs, and can also induce tumor cell apoptosis by enhancing the generation of reactive oxygen species (ROS). Therefore, there is an urgent need to study a scheme for combining ferrite hyperthermia with chemotherapy to treat breast cancer. Summary of the invention
[0005] In order to overcome the deficiencies of the above-mentioned prior art, the present invention provides a novel nanoparticle preparation with multifunctional diagnostic and therapeutic effects, namely, Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles and a preparation method thereof. The Ag2Fe2O4-TiO2 magnetic nanoparticle core is first prepared by ultrasonic chemical reduction, and then the egg yolk lecithin containing paclitaxel and the cholesterol substitute ginsenoside solution are coated on the surface of the core by ethanol injection to obtain Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles. The present invention combines magnetic nanomaterials with drug delivery systems, realizes multimodal treatment of tumors through photothermal effect, ROS generation and drug release, so that it has excellent biocompatibility and anti-cancer effects at the same time, and provides an innovative technical solution for the diagnosis and treatment of triple-negative breast cancer.
[0006] In order to achieve the above object, the technical solution adopted by the present invention is:
[0007] The first aspect of the present invention provides a method for preparing Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles, the method comprising the following steps:
[0008] S1, dissolving iron salt and silver nitrate in water, adding sodium hydroxide solution containing sodium borohydride NaBH4, generating a black precipitate through ultrasonic reaction, removing the supernatant, retaining the precipitate, adding ethanol and tetrabutyl titanate (TBOT) to the obtained precipitate, stirring thoroughly to promote surface titaniumization, washing and drying to obtain the Ag2Fe2O4-TiO2 magnetic nanoparticle core;
[0009] S2. Dissolve paclitaxel PTX, egg yolk lecithin and ginsenoside Rh2 in ethanol, and then inject them into the dextran solution containing Ag2Fe2O4-TiO2 magnetic nanoparticles by ethanol injection method. After stirring, a lipid nanoparticle solution is formed, and then Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles are prepared after dialysis purification and drying.
[0010] The present invention first uses iron salt, silver nitrate and tetrabutyl titanate as raw materials, and adopts ultrasonic chemical reduction method to prepare Ag2Fe2O4-TiO2 magnetic nanoparticle core; then, by ethanol injection method, egg yolk lecithin containing paclitaxel PTX and ginsenoside solution as cholesterol substitute are coated on the surface of the core to obtain Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles. The nanoparticles prepared by the present invention have small particle size and stable morphology, can significantly inhibit the invasion and migration of breast cancer cells at the cellular level, and can promote the generation of reactive oxygen species under photothermal stimulation, further synergistically induce cell apoptosis, and show strong cytotoxicity. In the animal model of breast cancer, the nanoparticles can combine the targeted photothermal therapy induced by near-infrared light NIR with immunotherapy, and then play a synergistic anti-tumor therapeutic effect, so that the inhibition rate of tumor volume reaches 82.56%, showing excellent anti-breast cancer effect and biosafety. The present invention provides a new and efficient response plan for the integrated diagnosis and treatment research of breast cancer.
[0011] Preferably, in S1, the iron salt is selected from one of ferric chloride hexahydrate, ferric nitrate nonahydrate and ferric sulfate nonahydrate.
[0012] Preferably, in S1, the molar ratio of the iron salt, silver nitrate and tetrabutyl titanate is 1-2:1-2:6-8.
[0013] Preferably, in S1, the concentration of the sodium hydroxide solution is 2-3 M, and the amount of sodium borohydride contained is 5-7 mmol, so as to control the precipitation rate and particle size distribution.
[0014] Preferably, in S1, the temperature of the ultrasonic reaction is room temperature and the time is 30 minutes.
[0015] Preferably, in S1, washing is performed by alcohol washing and water washing for several times until the magnetic nanoparticles are washed to a pH of 7.
[0016] Preferably, in S1, the drying temperature is 60-90°C and the drying time is 8-36 hours to obtain different crystal structures or magnetic properties.
[0017] Preferably, in S1, the ethanol added is anhydrous ethanol, and the volume of the ethanol added can be adjusted so that the volume ratio of ethanol to tetrabutyl titanate is 5-15:2-3, so as to optimize the titanium reaction conditions.
[0018] Preferably, in S2, the ratio of paclitaxel, ginsenoside Rh2 and egg yolk lecithin is 5:3-4:10 to ensure the optimal lipid composition.
[0019] Preferably, in S2, the volume ratio between the ethanol phase containing the lipid material (paclitaxel, egg yolk lecithin and ginsenoside Rh2) and the aqueous phase containing the magnetic core (magnetic nanoparticles) is 1:4.
[0020] Preferably, in S2, during the ethanol injection method, the stirring speed of ethanol injection is 700-900 rpm, the reaction temperature is 50-55 degrees Celsius, and the reaction time is 20-40 minutes.
[0021] Preferably, in S2, the dialysis bag used for the dialysis purification has a specification of 8-14KD and the dialysis time is 20-30h to remove free lipid components and drugs.
[0022] The second aspect of the present invention provides Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles prepared by the preparation method described in the first aspect.
[0023] The Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles prepared by the method of the present invention have a magnetic core and a lipid bilayer coated on the surface of the core, the core is composed of Ag2Fe2O4-TiO2, and the lipid layer contains paclitaxel and ginsenoside Rh2.
[0024] The third aspect of the present invention provides the use of the Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles described in the second aspect in the preparation of anti-breast cancer drugs.
[0025] Preferably, the breast cancer is triple-negative breast cancer.
[0026] The Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles prepared by the method of the present invention can be combined with immunotherapy for treatment through the photothermal effect induced by near-infrared light NIR, and have the effects of inhibiting tumor cell proliferation, promoting the generation of reactive oxygen species and inducing tumor cell apoptosis.
[0027] The fourth aspect of the present invention provides an anti-breast cancer drug, which uses the Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles described in the second aspect as a main active ingredient.
[0028] Preferably, the drug further comprises a pharmaceutically acceptable excipient.
[0029] More preferably, the excipients are diluents, adhesives, lubricants, disintegrants, solubilizers, stabilizers, etc. that can be used in the pharmaceutical field, as well as some pharmaceutical matrices; they can also be functional pharmaceutical excipients available in the pharmaceutical field, including surfactants, suspending agents, emulsifiers, and some new pharmaceutical polymer materials, such as cyclodextrin, chitosan, polylactic acid (PLA), polyglycolic acid-polylactic acid copolymer (PLGA), hyaluronic acid, etc.
[0030] Preferably, the dosage form of the drug includes tablets, granules, capsules, pills, sustained-release agents, oral liquid preparations, injection powders, and injections. The drug can be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically), i.e., it can be flexibly administered orally, subcutaneously, intramuscularly, and intravenously. If some drugs are unstable under gastric conditions, they can be prepared into enteric-coated tablets.
[0031] Compared with the prior art, the present invention has the following beneficial effects:
[0032] The invention discloses a method for preparing Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles. The method firstly prepares an Ag2Fe2O4-TiO2 magnetic core by ultrasonic chemical reduction method. The core has both magnetic responsiveness and photocatalytic activity, can generate efficient photothermal effect and ROS under near-infrared NIR light, and synergistically induces tumor cell apoptosis. Then, ginsenoside Rh2 is loaded on the surface of the core instead of cholesterol and paclitaxel PTX by ethanol injection method, and is coated with a lipid layer to form stable small-diameter nanoparticles. The lipid coating not only improves the stability and drug delivery efficiency of the nanoparticles, but also improves their in vivo metabolic characteristics.
[0033] The Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles prepared by the method of the present invention can significantly reduce the invasion and migration ability of breast cancer cells by inhibiting the expression of adhesion factors of tumor cells. At the same time, under NIR light, the nanoparticles can also generate local high temperature through the photothermal effect of the Ag2Fe2O4-TiO2 core, while inducing ROS generation, destroying the integrity of the tumor cell membrane and triggering cell apoptosis. Therefore, the Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles of the present invention show a significant tumor growth inhibition effect by combining photothermal and immunotherapy, with a high tumor inhibition rate, and can be combined with photothermal and chemotherapy to treat triple-negative breast cancer. In addition, the Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles of the present invention also have excellent biosafety, and no obvious toxic side effects were found during the treatment process, indicating that it has good clinical transformation potential.
[0034] The advantages and innovations of the Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles provided by the present invention are: (1) Multimodal treatment: integrating photothermal effect, ROS generation, chemotherapy and immunotherapy to provide a systematic tumor treatment solution. (2) Precision drug delivery: lipid nanoparticles improve the stability of drugs and the targeting of tumor sites, and reduce nonspecific toxicity. (3) Highly effective therapeutic effect: excellent anti-tumor activity is shown in both cell and animal models, and it is particularly suitable for the treatment of malignant tumors such as TNBC that lack targeted therapy.
[0035] In summary, the present invention provides a new type of Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles, which achieve efficient treatment of triple-negative breast cancer by combining photothermal, ROS generation and drug delivery, and provide a new solution for the multimodal diagnosis and treatment of cancer, which has important application prospects and social value. BRIEF DESCRIPTION OF THE DRAWINGS
[0036] Figure 1 is the hydrated particle size distribution diagram of Ag2Fe2O4-TiO2@Rh2-PTX NPs;
[0037] Figure 2 This is the Zeta potential analysis distribution diagram of Ag2Fe2O4-TiO2@Rh2-PTX NPs;
[0038] Figure 3 This is the photothermal conversion diagram of Ag2Fe2O4-TiO2@Rh2-PTX NPs under 808nm infrared laser irradiation;
[0039] Figure 4 is the toxicity diagram of Ag2Fe2O4-TiO2@Rh2-PTX NPs to 4T1 cells;
[0040] Figure 5 The graph of active oxygen production of Ag2Fe2O4-TiO2@Rh2-PTX NPs with and without light irradiation;
[0041] Figure 6 Figure 2 is the graph of tumor volume inhibition in 4T1 tumor-bearing Balb / c mice in different treatment groups. DETAILED DESCRIPTION
[0042] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in each embodiment of the present invention described below can be combined with each other as long as they do not conflict with each other.
[0043] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.
[0044] Example 1
[0045] This embodiment provides a method for preparing a novel Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticle, comprising the following steps:
[0046] (1) Preparation of Ag2Fe2O4-TiO2 magnetic nanoparticles
[0047] 2mmol of ferric nitrate nonahydrate and 2mmol of silver nitrate were dissolved in 20mL of deionized water, and 6mL of 2.5M sodium hydroxide (containing 250mg of sodium borohydride) solution was slowly added after ultrasonic mixing. After ultrasonic reaction at room temperature for 30 minutes to generate a black precipitate, the supernatant was removed by magnetic separation (silver ferrite was adsorbed by a strong magnet, and the liquid was poured to remove the supernatant, thereby retaining the magnetic silver ferrite), and the precipitate was retained (Ag2Fe2O4 was prepared in this step). 6mL of anhydrous ethanol and 2mL of tetrabutyl titanate were added to the obtained precipitate, and the mixture was fully stirred for 3 hours to promote surface titanization. The obtained precipitate was then washed 3-6 times with anhydrous ethanol and deionized water respectively until the pH of the washing solution was close to neutral. Finally, the obtained washed product was placed in an oven at 80°C and dried for 12 hours to obtain the Ag2Fe2O4-TiO2 magnetic nanoparticle core.
[0048] (2) Preparation of Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles
[0049] 10 mg of paclitaxel PTX, 20 mg of egg yolk lecithin and 6 mg of ginsenoside Rh2 were dissolved in 1.5 mL of anhydrous ethanol and stirred for 10-15 minutes to form a uniform solution; then the obtained lipid and drug solution were injected dropwise into 6 mL of a dextran aqueous solution containing Ag2Fe2O4-TiO2 magnetic nanoparticles (the concentration of the magnetic nanoparticles was 4 μg / mL, and the concentration of the dextran aqueous solution was 2%) by the ethanol injection method, that is, the lipids and drugs in the ethanol phase were injected into the bottom of the aqueous phase using a syringe, and the mixture was pushed dropwise while stirring, and then stirred at 55°C and 900 rpm for 30 minutes to ensure that the drugs and lipids were evenly coated on the surface of the magnetic core to form a lipid nanoparticle solution. The resulting solution was placed in a 10KD dialysis bag and dialyzed with deionized water for 24 hours, during which the dialysate was replaced regularly to ensure the removal of unreacted solvents and free components. Finally, the dialyzed product was freeze-dried to prepare Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles, which were stored in a dry environment to avoid moisture absorption.
[0050] Comparative Example 1
[0051] This comparative example provides a method for preparing Ag2Fe2O4-TiO2@Chol-PTX lipid nanoparticles, and its preparation process is the same as that of Example 1, except that in step (2), 10 mg of paclitaxel PTX, 20 mg of egg yolk lecithin and 6 mg of cholesterol (Chol) are dissolved in anhydrous ethanol and then injected into the dextran solution of magnetic nanoparticles by ethanol injection method.
[0052] Experimental Example 1: Characterization of Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles
[0053] (1) Particle size and polydispersity (PDI)
[0054] The sample Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles to be tested were placed in a cuvette, and the particle size of the Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles was measured by dynamic light scattering (DLS) to be 144.26 nm, and the polydispersity index (PDI) was 0.148, which showed that the particle size distribution of the preparation was uniform ( Figure 1 Finally, the Zeta potential of Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles was measured by dynamic light scattering (DLS) to be -18 mV ( Figure 2 ).
[0055] (2) Ag and Fe content
[0056] The preparation was digested by microwave (2 mg of sample and 10 mL of aqueous solution of Rh2O4 were added into a high-pressure sealed digestion tank, heated to 180°C in a microwave digester and maintained for 30 minutes) and the elemental contents of Ag2Fe2O4 and Ag2Fe2O4-TiO2@Rh2-PTX in 1 mg of freeze-dried liposome preparation were determined by inductively coupled plasma optical emission spectroscopy (ICP-OES): Ag2Fe2O4-TiO2@Rh2-PTX: Fe content was 11.15 μg, and Ag content was 1.6 μg; Ag2Fe2O4: Fe content was 221 μg, and Ag content was 50.5 μg.
[0057] (3) Photothermal effect
[0058] Under 808nm near-infrared laser irradiation, 2mg / mL Ag2Fe2O4 and 2mg / mL Ag2Fe2O4-TiO2 heated to 50℃ and 60℃ respectively within 10min, proving that the addition of TiO2 significantly improved the photothermal conversion efficiency. At the same time, the 2mg / mL Ag2Fe2O4-TiO2@Rh2-PTX formulation solution also heated to 58℃ within 10 minutes, proving that it has significant photothermal conversion ability ( Figure 3 ).
[0059] Experimental Example 2: Verification of the function of Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles through in vitro experiments
[0060] (1) Cytotoxicity assay
[0061] In vitro cytotoxicity experiments were performed using 4T1 breast cancer cells, and the in vitro cytotoxicity of the preparations was evaluated by the CCK-8 method. 4 Breast cancer cell line (4T1) was seeded into 96-well plates at a standard of 0.1 mL of culture medium (10% FBS in DMEM) per well. After 24 h, the cells were plated in paclitaxel, Ag2Fe2O4-TiO2, Ag2Fe2O4-TiO2@Chol-PTX, Ag2Fe2O4-TiO2@Rh2-PTX, Ag2Fe2O4-TiO2@Rh2-PTX-NIR (2 W / cm 2 , irradiation 4min) added different concentrations (according to PTX, 1.5625, 3.125, 6.25, 12.5, 25, 50, 100 μg / mL) of the agent (the agent was diluted with PBS) and incubated the cells. After incubation for 24 hours under cell culture conditions (37°C, 5% CO2), 10 μL of cck-8 reagent was added to each well and incubated for another hour at 37°C. Finally, the absorbance at 450nm was measured using an ELISA reader.
[0062] The results show that Figure 4 ): The toxicity of Ag2Fe2O4-TiO2@Rh2-PTX and Ag2Fe2O4-TiO2@Chol-PTX was significantly higher than that of paclitaxel API (PTX), and the cytotoxicity of Ag2Fe2O4-TiO2@Rh2-PTX was better than that of Ag2Fe2O4-TiO2@Chol-PTX, which proved that ginsenoside Rh2 improved the anticancer activity of the preparation. In addition, the photothermal group had the strongest cytotoxicity, indicating that photothermal therapy further enhanced the anti-tumor effect.
[0063] (2) Detection of reactive oxygen species (ROS)
[0064] 1 × 10 cells were seeded in 6-well cell culture plates. 6 / mL 4T1 cells were incubated in a cell culture incubator for 24h, and then H2DCFDA fluorescent probe was added and incubated at 37℃ for 20min. After that, it was fully washed three times with serum-free DMEM medium to remove the unloaded fluorescent probe. Then, 1mL PTX solution, or Ag2Fe2O4-TiO2@Chol-PTX solution, or Ag2Fe2O4-TiO2@Rh2-PTX solution, or blank control solution was added to each well. The average concentration of each solution was 10μg / mL in terms of PTX. The blank control was blank serum-free DMEM medium. After administration, it was incubated for 2h, washed three times with serum-free DMEM medium, and then an 808nm laser emitter was used to perform photothermal treatment (2W / cm 2 , irradiated for 4 min), and finally the fluorescence intensity was detected on a fluorescence microplate reader at an excitation wavelength of 488 nm and an emission wavelength of 525 nm.
[0065] The results show that Figure 5 ): The ROS generation in the Ag2Fe2O4-TiO2@Rh2-PTX-NIR group was the highest, indicating that it had the strongest ability to induce oxidative stress response through the photothermal effect, and both Ag2Fe2O4 and light NIR promoted the production of reactive oxygen species.
[0066] Experimental Example 3: Verification of the function of Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles through in vivo experiments
[0067] (1) Breast cancer animal model
[0068] BALB / c mice weighing about 20 g were used to establish a breast cancer model (subcutaneous injection of 1×10 6 4T1 breast cancer cells were grown in the mammary fat pad. When the tumor volume reached 90-100mm 3The mice were randomly divided into 6 groups: normal saline group (model control group), NIR group, paclitaxel (PTX) group, Ag2Fe2O4-TiO2@Chol-PTX group, Ag2Fe2O4-TiO2@Rh2-PTX group, Ag2Fe2O4-TiO2@Rh2-PTX-NIR phototherapy group
[0069] The tumor volume and mouse weight were measured every two days in each group, and the treatment cycle was 21 days. The tumor volume was calculated as follows: tumor volume = length × width × width / 2.
[0070] (2) Anti-tumor experiments
[0071] In the triple-negative breast cancer mouse model, the drugs were injected into the tail vein according to the normal saline group (model control group), NIR group, paclitaxel (PTX) group, Ag2Fe2O4-TiO2@Chol-PTX group, Ag2Fe2O4-TiO2@Rh2-PTX group, and Ag2Fe2O4-TiO2@Rh2-PTX-NIR group. When the tumor volume reached 90-100mm 3 The drug was administered on day 0 and every two days at a concentration of 10 mg / kg (in terms of PTX) for a treatment period of 21 days. 2 ) induction, that is, 4 minutes of light exposure was performed 1 hour after each preparation administration.
[0072] The experimental results show that ( Figure 6 ), the tumor inhibition rate of the Ag2Fe2O4-TiO2@Rh2-PTX group was 60.17%, while that of the Ag2Fe2O4-TiO2@Rh2-PTX-NIR phototherapy group could reach 82.56%, and there was no significant change in the body weight of the mice, indicating that Ag2Fe2O4-TiO2@Rh2-PTX combined with photothermal therapy has an excellent effect in the treatment of triple-negative breast cancer.
[0073] In summary, the present invention combines magnetic nanomaterials (Ag2Fe2O4-TiO2) with drug delivery systems (containing paclitaxel, egg yolk phosphatidylcholine and ginsenosides) to prepare a new type of Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles, which achieve efficient treatment of triple-negative breast cancer by combining photothermal, ROS generation and drug delivery, providing a new solution for multimodal diagnosis and treatment of cancer, and has important application prospects and social value.
[0074] The embodiments of the present invention are described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions and variations of these embodiments are made without departing from the principles and spirit of the present invention, and still fall within the protection scope of the present invention.
Claims
1. A method for preparing Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles, characterized in that: The following steps are involved: S1. Dissolve iron salt and silver nitrate in water, add sodium hydroxide solution containing sodium borohydride, generate black precipitate through ultrasonic reaction, remove supernatant, retain precipitate, add ethanol and tetrabutyl titanate to the precipitate, stir well to promote surface titaniumization, wash and dry to obtain Ag2Fe2O4-TiO2 magnetic nanoparticle core; S2. Paclitaxel, egg yolk lecithin and ginsenoside were dissolved in ethanol, and then injected into the dextran solution containing Ag2Fe2O4-TiO2 magnetic nanoparticles by ethanol injection method. After stirring, lipid nanoparticle solution was formed, and then Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles were prepared after dialysis purification and drying.
2. The method for preparing Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles according to claim 1, characterized in that: In S1, the iron salt is selected from one of ferric chloride hexahydrate, ferric nitrate nonahydrate, and ferric sulfate nonahydrate; the molar ratio of the iron salt, silver nitrate, and tetrabutyl titanate is 1-2:1-2:6-8, and the volume ratio of ethanol to tetrabutyl titanate is 5-15:2-3.
3. The method for preparing Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles according to claim 1, characterized in that: In S1, the concentration of the sodium hydroxide solution is 2-3M, and the amount of sodium borohydride contained is 5-7mmol.
4. The method for preparing Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles according to claim 1, characterized in that: In S1, the temperature of the ultrasonic reaction is room temperature and the time is 20-40 minutes.
5. The method for preparing Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles according to claim 1, characterized in that: In S1, washing is performed by alcohol washing and water washing for several times until the magnetic nanoparticles are washed to a pH of 7; the drying temperature is 60-90° C., and the drying time is 8-36 hours.
6. The method for preparing Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles according to claim 1, characterized in that: In S2, the ratio of paclitaxel, ginsenoside Rh2 and egg yolk lecithin is 5:3-4:10; the volume ratio between the ethanol phase containing paclitaxel, egg yolk lecithin and ginsenoside and the aqueous phase containing the magnetic core is 1:
4.
7. The method for preparing Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles according to claim 1, characterized in that: In S2, during the ethanol injection method, the stirring speed of ethanol injection is 700-900 rpm, the reaction temperature is 50-55 degrees Celsius, and the reaction time is 20-40 minutes; the specification of the dialysis bag used for dialysis purification is 8-14KD, and the dialysis time is 20-30h.
8. Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles prepared by the preparation method according to any one of claims 1 to 7.
9. Use of the Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles according to claim 8 in the preparation of anti-breast cancer drugs.
10. An anti-breast cancer drug, characterized in that: The drug uses the Ag2Fe2O4-TiO2@Rh2-PTX lipid nanoparticles described in claim 8 as the main active ingredient.