A lanthanide metal-organic framework composite material and its preparation method and application

Abstract

The present invention discloses a lanthanide metal organic framework composite material and its preparation method and application, which belongs to the technical field of targeted material preparation. The preparation method of the lanthanide metal organic framework composite material of the present invention comprises the following steps: mixing and reacting a lanthanide metal salt and an organic ligand in the form of a solution to obtain an organic framework material; mixing and reacting triphenylphosphine-modified doxorubicin and an organic framework material in the form of a solution to obtain an organic framework material carrying TPP-DOX; mixing and reacting a polysaccharide having a function of targeting CD44 receptors on the surface of cancer cells and an organic framework material carrying TPP-DOX in the form of a solution to obtain a lanthanide metal organic framework composite material. The lanthanide metal organic framework composite material prepared by the present invention can accurately identify and locate tumor tissue without adversely affecting surrounding normal tissues.

Description

Technical Field

[0001] The present invention relates to the technical field of targeted material preparation, and in particular to a lanthanide metal organic framework composite material and a preparation method and application thereof. Background Art

[0002] By the time most cancers are diagnosed, they are typically in the late stages, well past the optimal time for surgical treatment. Therefore, early diagnosis and treatment are crucial. Traditional cancer treatments, including surgery, radiotherapy, and chemotherapy, are significantly limited by their targeting. Therefore, actively developing new, targeted, and precision-guided therapies is both a challenging and exciting area of ​​research.

[0003] Recent research has explored a variety of novel materials for the treatment of malignant tumors, including metal-organic frameworks, biodegradable polymers, nanoparticles, and nanomagnetic materials. The research and application of these materials have provided new insights and approaches for tumor therapy, potentially leading to more effective and safer clinical cancer treatment strategies. Lanthanide metal-organic frameworks (Ln-MOFs) are a class of porous crystalline materials composed of lanthanide metal ions and organic ligands. These materials are composed of metal ions (such as lanthanum, erbium, and neodymium) and organic ligands (such as carboxylic acids and pyridines) linked by coordinated chemical bonds, forming a lattice with a regular pore structure. Ln-MOFs are highly tunable and diverse, and their pore size, pore structure, and surface functionalization can be manipulated by selecting different metal ions and organic ligands. Due to their unique structural characteristics, Ln-MOFs have broad application prospects in gas adsorption and separation, catalysis, drug delivery, optics, and magnetism. As a novel material, Ln-MOFs have shown potential application value in tumor treatment, including drug delivery, photothermal therapy, photodynamic therapy, and tumor imaging. However, their biocompatibility and toxicity in vivo remain key issues. Some Ln-MOFs may induce immune responses or toxic effects, limiting their clinical application. Although Ln-MOFs can be functionalized or modified to achieve targeted delivery to tumors, their targeting performance needs further improvement. Ensuring that Ln-MOFs can accurately identify and locate tumor tissue without adversely affecting surrounding normal tissues is a challenge. Summary of the Invention

[0004] The purpose of the present invention is to provide a lanthanide metal organic framework composite material and a preparation method and application thereof, so as to solve the problems existing in the above-mentioned prior art.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] One of the technical solutions of the present invention is a method for preparing a lanthanide metal-organic framework composite material, comprising the following steps:

[0007] The lanthanide metal salt and the organic ligand are mixed and reacted in the form of a solution to obtain an organic framework material;

[0008] The triphenylphosphine-modified doxorubicin (TPP-DOX) and the organic framework material are mixed and reacted in the form of a solution to obtain an organic framework material carrying TPP-DOX;

[0009] The polysaccharide having the function of targeting the CD44 receptor on the surface of cancer cells and the organic framework material carrying TPP-DOX are mixed and reacted in the form of a solution to obtain the lanthanide metal-organic framework composite material.

[0010] Furthermore, the lanthanide metal salt includes cerium nitrate and europium nitrate;

[0011] The organic ligand includes pyridine-2,6-dicarboxylic acid (DPA);

[0012] The molar ratio of the cerium nitrate, europium nitrate and pyridine-2,6-dicarboxylic acid is 0.125:0.125:(0.1-0.5).

[0013] Furthermore, the lanthanide metal salt is mixed with the organic ligand twice.

[0014] Mixing the lanthanide metal salt with the organic ligand twice is more conducive to the dissolution of the raw materials and the synthesis of the materials.

[0015] Furthermore, the mass ratio of the organic framework material to triphenylphosphine-modified doxorubicin is 20:4.

[0016] Furthermore, the time for mixing the triphenylphosphine-modified doxorubicin and the organic framework material in the form of a solution is 6 hours.

[0017] Furthermore, the mass ratio of the organic skeleton material carrying TPP-DOX and the polysaccharide having the function of targeting the CD44 receptor on the surface of cancer cells is 1:1.

[0018] Furthermore, the polysaccharide having the function of targeting the CD44 receptor on the surface of cancer cells and the organic framework material carrying TPP-DOX are mixed in the form of a solution and reacted for 24 hours;

[0019] The polysaccharide having the function of targeting CD44 receptor on the surface of cancer cells comprises hyaluronic acid (HA).

[0020] The organic framework material (Eu / Ce DPA) generated in this invention using pyridine-2,6-dicarboxylic acid (DPA) combined with the metal ions cerium and europium (Eu / Ce) is a special type of bimetallic lanthanide organic framework. As a new material, it has shown potential application value in drug delivery and tumor imaging in tumor treatment. Triphenylphosphine-modified doxorubicin (TPP-DOX) is a positively charged drug delivery system with the ability to target negatively charged mitochondria; hyaluronic acid (HA) is a polysaccharide that can target the CD44 receptor on the surface of cancer cells. The new lanthanide metal-organic framework composite (Eu / Ce DPA@TPP-DOX@HA) synthesized using Eu / Ce DPA, TPP-DOX, and HA has multiple functions, including targeted delivery, drug protection and controlled release, and optical labeling, with good biocompatibility and minimal side effects.

[0021] Eu / Ce DPA is a special type of lanthanide metal-organic framework (MOF) that contains two different metal ions (Eu / Ce) simultaneously. This bimetallic structure can introduce more active sites to the MOF or modulate its surface chemical properties, thereby improving its performance in gas adsorption, catalysis, separation, and other fields. Using Eu / Ce DPA as the target material skeleton, TPP-DOX was loaded onto the structure to synthesize Eu / Ce DPA@TPP-DOX. This improved the in vivo stability and tumor targeting of Eu / Ce DPA@TPP-DOX. Attaching HA to the surface of Eu / Ce-DPA@TPP-DOX enhanced tumor imaging and targeting, resulting in the final target-sensitive and specific tumor-killing composite material, Eu / Ce PA@TPP-DOX@HA.

[0022] The second technical solution of the present invention: a lanthanide metal-organic framework composite material prepared by the above preparation method.

[0023] The third technical solution of the present invention: an application of the above-mentioned lanthanide metal-organic framework composite material in the preparation of drugs for targeted tumor treatment.

[0024] The present invention discloses the following technical effects:

[0025] The lanthanide metal-organic framework composite material prepared by the present invention can accurately identify and locate tumor tissues without causing adverse effects on surrounding normal tissues.

[0026] The lanthanide metal-organic framework composite material prepared by the invention has multiple functions such as targeted delivery, drug protection and controlled release, optical labeling, etc., and has good biocompatibility and small side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0028] Figure 1 Characterization results of Eu / Ce DPA prepared in Example 1, where a is a TEM image and b is the EDS analysis result;

[0029] Figure 2 Characterization results of Eu / Ce DPA@TPP-DOX@HA prepared in Example 1, where a is a TEM image and b is the EDS analysis result;

[0030] Figure 3 IR spectra of pyridine-2,6-dicarboxylic acid (DPA) and Eu / Ce DPA (MOF) prepared in Example 1;

[0031] Figure 4 This is the high-resolution spectrum of Eu / Ce DPA@TPP-DOX@HA prepared in Example 1, where a is the full spectrum, b is the C1s spectrum, c is the N1s spectrum, d is the O1s spectrum, e is the Ce 3d spectrum, f is the Eu3d spectrum, and g is the P 2p spectrum;

[0032] Figure 5 X-ray diffraction spectra of Eu / Ce DPA prepared in Example 1 and Eu / Ce DPA@TPP-DOX@HA prepared in Example 1;

[0033] Figure 6 These are the ultraviolet absorption spectra of the raw material TPP-DOX used in Example 1, the Eu / Ce DPA prepared in Example 1, and the Eu / Ce DPA@TPP-DOX@HA prepared in Example 1;

[0034] Figure 7 Zeta potential diagrams of Eu / Ce DPA(MOF) prepared in Example 1, TPP-DOX used as a raw material in Example 1, HA used as a raw material in Example 1, Eu / Ce DPA@TPP-DOX(MT) prepared in Example 1, and Eu / Ce DPA@TPP-DOX@HA(MTH) prepared in Example 1;

[0035] Figure 8 The fluorescence emission spectra of Eu / Ce DPA@TPP-DOX@HA prepared in Example 1 under different excitation lights;

[0036] Figure 9 The active oxygen generation capacity of Eu / Ce DPA (MOF) prepared in Example 1 and Eu / Ce DPA@TPP-DOX@HA (MTH) prepared in Example 1;

[0037] Figure 10 The expression of CD44 in normal thyroid cells NTHY-ORI-3-1 and anaplastic thyroid cancer cells CAL-62;

[0038] Figure 11 The killing ability of Eu / Ce DPA, Eu / Ce DPA@TPP-DOX and Eu / Ce DPA@TPP-DOX@HA prepared in Example 1 on cancer cells (CAL-62 cells) and normal tissue cells (NTHY-ORI-3-1 cells), where a is Eu / CeDPA, b is Eu / Ce DPA@TPP-DOX, and c is Eu / Ce DPA@TPP-DOX@HA;

[0039] Figure 12 The fluorescence intensity of Eu / Ce DPA@TPP-DOX@HA prepared in Example 1 after co-incubation with normal thyroid cells NTHY-ORI-3-1 and anaplastic thyroid cancer cells CAL-62. DETAILED DESCRIPTION

[0040] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0041] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between any intermediate value within a stated value or stated range and any other stated value or intermediate value within the stated range is also encompassed by the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0042] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0043] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be exemplary only.

[0044] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0045] Example 1

[0046] A method for preparing a lanthanide metal-organic framework composite material:

[0047] (1) Synthesis of Eu / Ce DPA (organic framework material):

[0048] (1) Cerium nitrate hexahydrate (Ce(NO3)3·6H2O, 0.125 mmol, 0.0542 g) and europium nitrate hexahydrate (Eu(NO3)3·6H2O, 0.125 mmol, 0.0557 g) were dissolved in 5 mL of primary water to obtain a metal ion solution;

[0049] The organic ligand pyridine-2,6-dicarboxylic acid (DPA, 0.5 mmol, 0.0836 g) was dissolved in 15 mL of an ethanol solution containing triethylamine (1 mmol, 134 μL) to obtain an organic ligand solution.

[0050] (2) 100 μL of the above metal ion solution was added dropwise to the organic ligand solution, and ultrasonic reaction was performed for 10 min (ultrasonic power was 40 W). Then, the remaining metal ion solution was added dropwise to the mixed solution, and ultrasonic reaction was continued for 60 min. The reaction system gradually produced a yellow precipitate, which was centrifuged (10,000 rpm, 3 min), washed twice with ethanol, and freeze-dried overnight to obtain Eu / CeDPA, a yellow solid powder.

[0051] (2) Synthesis of Eu / Ce DPA@TPP-DOX:

[0052] (1) Eu / Ce DPA powder (20 mg) was ultrasonically dispersed in 16 mL of Tris-HCl buffer (10 mM, pH 8.5) to form a clear Eu / Ce DPA solution;

[0053] TPP-DOX (4 mg) was dissolved in 4 mL of Tris-HCl buffer (10 mM, pH 8.5) to form an orange-red transparent TPP-DOX solution.

[0054] (2) The above TPP-DOX solution was added dropwise to the Eu / Ce DPA solution and stirred at room temperature for 6 h under a magnetic stirring speed of 500 rpm until the mixed solution became a purple transparent solution. After centrifugation (12,000 rpm, 5 min), washed twice with ethanol, and freeze-dried overnight, Eu / Ce DPA@TPP-DOX (organic framework material carrying TPP-DOX) was obtained as a purple solid powder.

[0055] (III) Synthesis of lanthanide metal-organic framework composites:

[0056] (1) Eu / Ce DPA@TPP-DOX powder (20 mg) was ultrasonically dispersed in 19 mL of ethanol solution to obtain Eu / CeDPA@TPP-DOX dispersion;

[0057] Hyaluronic acid (HA, 20 mg) was dissolved in 1 mL of first-grade water to obtain a HA solution.

[0058] (2) The above HA solution was added dropwise to the Eu / Ce DPA@TPP-DOX dispersion and stirred at room temperature for 24 h under a magnetic stirring speed of 500 rpm. After centrifugation (12,000 rpm, 5 min), washed twice with ethanol, and freeze-dried overnight, Eu / Ce DPA@TPP-DOX@HA (lanthanide metal-organic framework composite material) was obtained as a purple solid powder.

[0059] Effect Example 1

[0060] (1) The Eu / Ce DPA prepared in Example 1 was characterized. The results are shown in Figure 1 , Figure 1 a is the TEM image, and b is the EDS analysis result.

[0061] from Figure 1 It can be seen that the Eu / Ce DPA prepared in Example 1 exhibits a flaky nanostructure, and its EDS results show that the Eu / Ce DPA contains Eu, Ce and N elements and is evenly distributed with an average size of about 200 nm.

[0062] (2) The Eu / Ce DPA@TPP-DOX@HA prepared in Example 1 was characterized. The results are shown in Figure 2 , Figure 2 a is the TEM image, and b is the EDS analysis result.

[0063] from Figure 2 It can be seen that the shape of Eu / Ce DPA@TPP-DOX@HA is nanoclusters. Its EDS results show that it contains not only Eu, Ce and N elements in Eu / Ce DPA, but also P element unique to TPP-DOX. The elements are evenly distributed with an average size of about 250 nm.

[0064] (3) The raw material pyridine-2,6-dicarboxylic acid (DPA) used in Example 1 and the Eu / Ce DPA (MOF) prepared in Example 1 were analyzed by Fourier transform infrared spectroscopy. The results are shown in Figure 3 .

[0065] from Figure 3 It can be seen that the characteristic asymmetric stretching vibration peak of DPA carbonyl is 1704.8 cm -1 The synthesized Eu / Ce DPA red-shifted to 1617.3 cm -1 , demonstrating the formation of a coordination bond between the carbonyl group and the lanthanide ion.

[0066] (4) The high-resolution spectrum of Eu / Ce DPA@TPP-DOX@HA prepared in Example 1 is shown in Figure 4 . Figure 4 In the figure, a is the full spectrum, b is the C1s spectrum, c is the N1s spectrum, d is the O1s spectrum, e is the Ce 3d spectrum, f is the Eu 3d spectrum, and g is the P 2p spectrum.

[0067] Eu / Ce DPA@TPP-DOX@HA has diffraction peaks of Eu 3d, Ce 3d, P 2p, O 1s, N 1s and C1s, among which the C 1s spectrum ( Figure 4 Figure b) shows three characteristic peaks of C=O (288.1eV), CH (285.2eV), and CC (284.4eV), and the Eu 3d spectrum ( Figure 4 Figure f) shows that there are two characteristic peaks of Eu 3d 5 / 2 (1134.9eV) and Eu 3d 3 / 2 (1164.6eV), and there is a characteristic satellite peak of Eu 3d (1143.1eV) between the two diffraction peaks. After loading TPP-DOX, the unique characteristic peak of P element appears at 190.4eV. Eu, Ce, P, O, N and C elements are present in Eu / Ce DPA@TPPDOX@HA.

[0068] Furthermore, inductively coupled plasma optical emission spectrometry (ICP-OES) was used to characterize the Eu 3+ and Ce 3+ The content was measured. In Eu / Ce DPA (100 μg / mL), the Eu element content was 18.35 ppm (mg / g), and the Ce element content was 18.11 ppm (mg / g). The ratio of the two was about 1, which was consistent with the Eu element content during synthesis. 3+ / Ce 3+ The feed ratio is consistent. In Eu / Ce DPA@TPP-DOX@HA (100μg / mL), the Eu element content is 6.95ppm (mg / g), and the Ce element content is 5.49ppm (mg / g). The ratio of the two is also about 1, indicating that the synthesized material still contains Eu. 3+ and Ce 3+ .

[0069] (5) X-ray diffraction spectra of Eu / Ce DPA prepared in Example 1 and Eu / Ce DPA@TPP-DOX@HA prepared in Example 1 are shown in Figure 5 .

[0070] The UV absorption spectra of the raw material TPP-DOX used in Example 1, the Eu / Ce DPA prepared in Example 1, and the Eu / CeDPA@TPP-DOX@HA prepared in Example 1 are shown in FIG. Figure 6 .

[0071] Zeta potential diagrams of Eu / Ce DPA (MOF) prepared in Example 1, TPP-DOX used as raw material in Example 1, HA used as raw material in Example 1, Eu / Ce DPA@TPP-DOX (MT) prepared in Example 1, and Eu / Ce DPA@TPP-DOX@HA (MTH) prepared in Example 1 are shown in FIG. Figure 7 .

[0072] In a Tris-HCl buffer solution (10 mM, pH 8.5), negatively charged Eu / Ce DPA and positively charged TPP-DOX interacted electrostatically to synthesize the positively charged Eu / Ce DPA@TPP-DOX. Negatively charged HA was also coated on the surface of Eu / Ce DPA@TPP-DOX through electrostatic interactions. Ultraviolet absorption spectroscopy confirmed the successful loading of TPP-DOX and the successful coating of HA. After TPP-DOX loading and HA coating, the UV absorption spectrum was significantly improved.

[0073] (6) Fluorescence emission spectra of Eu / Ce DPA@TPP-DOX@HA prepared in Example 1 under different excitation light. Figure 8 .

[0074] (7) The active oxygen generation capacity of Eu / Ce DPA (MOF) prepared in Example 1 and Eu / Ce DPA@TPP-DOX@HA (MTH) prepared in Example 1 was measured (the results are shown in Figure 9 ), the determination method is as follows:

[0075] Eu / Ce DPA and Eu / Ce DPA@TPP-DOX@HA were reacted with hydrogen peroxide and / or TMB, respectively, and the UV absorption of the reaction system was measured to verify their enzyme-like activity.

[0076] Taking Eu / Ce DPA as an example, the specific experimental scheme is as follows: (1) Eu / Ce DPA (the concentration of Eu / Ce DPA in PBS is 100 μg / mL) and TMB (the concentration of TMB in PBS is 1 mg / mL) were added to PBS (10 mM, pH 5.5), and the reaction was carried out on a constant temperature shaker at a shaking rate of 300 rpm and reaction conditions of 37°C for 30 min. The ultraviolet absorption spectrum of the reaction system was measured using an enzyme-linked microplate reader (MOF+TMB group); the control group was without Eu / Ce DPA, and the ultraviolet absorption spectrum of the reaction system was measured using an enzyme-linked microplate reader (TMB group).

[0077] (2) Eu / Ce DPA (the concentration of Eu / Ce DPA in PBS was 100 μg / mL), hydrogen peroxide (the concentration of hydrogen peroxide in PBS was 100 μM), and TMB (the concentration of TMB in PBS was 1 mg / mL) were added to PBS (10 mM, pH 5.5). The mixture was reacted on a thermostat at 300 rpm and 37°C for 30 min. The UV absorption spectrum of the reaction system was measured using a microplate reader (MOF+TMB+H2O2 group). The control group did not add Eu / Ce DPA, and the UV absorption spectrum of the reaction system was measured using a microplate reader (TMB+H2O2 group).

[0078] (3) Eu / Ce DPA@TPP-DOX@HA (the concentration of Eu / Ce DPA@TPP-DOX@HA in PBS was 100 μg / mL) and TMB (the concentration of TMB in PBS was 1 mg / mL) were added to PBS (10 mM, pH 5.5). The mixture was reacted at 37°C on a thermostat at a shaking rate of 300 rpm for 30 min. The UV absorption spectrum of the reaction system was measured using a microplate reader (MTH+TMB group).

[0079] (4) Eu / Ce DPA@TPP-DOX@HA (the concentration of Eu / Ce DPA@TPP-DOX@HA in PBS was 100 μg / mL), hydrogen peroxide (the concentration of hydrogen peroxide in PBS was 100 μM), and TMB (the concentration of TMB in PBS was 1 mg / mL) were added to PBS (10 mM, pH 5.5). The mixture was reacted on a thermostat at a shaking rate of 300 rpm and at 37°C for 30 min. The UV absorption spectrum of the reaction system was measured using a microplate reader (MTH+TMB+H2O2 group).

[0080] Effect Example 2

[0081] To verify whether CD44 is highly expressed in anaplastic thyroid cancer cells CAL-62, flow cytometry was used to detect the CD44 levels in normal thyroid cells NTHY-ORI-3-1 and anaplastic thyroid cancer CAL-62 cells. Figure 10 .

[0082] The specific verification method is as follows:

[0083] (1) NTHY-ORI-3-1 and CAL-62 cells were cultured to a certain concentration (1-2×10 6 / mL), the material (Eu / Ce DPA@TPP-DOX@HA prepared in Example 1) was prepared into material solutions with concentrations of 1000 mg / mL, 500 mg / mL, 250 mg / mL, and 150 mg / mL, respectively, and the material solutions of different concentrations were mixed with cells (the volume ratio of material solution to cells was 200 μL:1.5 mL) to obtain cell suspensions;

[0084] (2) Distribute the cell suspension into individual centrifuge tubes, each containing approximately 10 6 Add CD44 antibody to the cell suspension to allow it to bind to CD44 on the cell surface, and incubate the cell and antibody mixture at 4°C;

[0085] (3) Wash the cells with PBS to remove unbound antibodies;

[0086] (4) The labeled and fixed cell suspension is placed in a flow cytometer sample tube, and the fluorescence intensity of the fluorescent marker in the cell suspension is detected using a flow cytometer. The fluorescence intensity of the CD44 antibody is detected using the PE channel.

[0087] from Figure 10 It can be seen that the CD44 expression level in anaplastic thyroid cancer CAL-62 cells is significantly higher than that in normal thyroid cells NTHY-ORI-3-1, indicating that the material can better target cancer cells without combining with normal cells.

[0088] Effect Example 3

[0089] CCK8 experiments verified the killing ability of Eu / Ce DPA, Eu / Ce DPA@TPP-DOX and Eu / Ce DPA@TPP-DOX@HA prepared in Example 1 at different concentrations on cancer cells (CAL-62 cells) and normal tissue cells (NTHY-ORI-3-1 cells). Figure 11 . Figure 11 In the figure, a is Eu / Ce DPA, b is Eu / Ce DPA@TPP-DOX, and c is Eu / Ce DPA@TPP-DOX@HA.

[0090] The specific verification method is as follows:

[0091] (1) Cell seeding: Cells in logarithmic growth state were seeded into 96-well plates at an appropriate density (5000 cells per well) and pre-cultured in an incubator for 24 h;

[0092] (2) Sample treatment: The material solution (Eu / Ce DPA, Eu / Ce DPA@TPP-DOX, or Eu / Ce DPA@TPP-DOX@HA prepared in Example 1) was prepared into material solutions with concentrations of 1000 mg / mL, 500 mg / mL, 250 mg / mL, and 150 mg / mL, respectively (prepared using cell culture medium). 100 μL of material solutions of different concentrations were added to a 96-well plate seeded with cells, and the wells without material solution and only with cell culture medium were used as blank controls.

[0093] (3) Incubation: Place the 96-well plate in an incubator and incubate for 24 hours to allow the cells to fully contact with the material.

[0094] (4) Adding CCK-8 reagent: After the incubation, add CCK-8 reagent to each well of the 96-well plate at 10 μL / well;

[0095] (5) Incubation: Place the 96-well plate back into the incubator and continue incubating for 4 h to allow the cells to absorb the CCK-8 reagent.

[0096] (6) Absorbance measurement: Place the 96-well plate in the wells to be measured and set the absorption wavelength to 450 nm.

[0097] (7) Data analysis: Based on the absorbance measurement results, draw the cell activity curve and calculate the relative cell activity of each sample.

[0098] Effect Example 4

[0099] After incubating NTHY-ORI-3-1 normal thyroid cells and CAL-62 anaplastic thyroid cancer cells with Eu / CeDPA@TPP-DOX@HA solution for 6 h, the cell fluorescence intensity was detected by flow cytometry. Figure 12 .

[0100] The specific verification method is as follows:

[0101] (1) NTHY-ORI-3-1 and CAL-62 cells were cultured to an appropriate concentration (1-2×10 6 / mL), the material (Eu / Ce DPA@TPP-DOX@HA prepared in Example 1) was prepared into material solutions with concentrations of 1000 mg / mL, 500 mg / mL, 250 mg / mL, and 150 mg / mL, respectively. The material solutions of different concentrations were mixed with cells (the volume ratio of material solution to cells was 200 μL:1.5 mL).

[0102] (2) Distribute the cell suspension into individual centrifuge tubes, each containing approximately 10 6 Cells were incubated with the material solution for 6 h.

[0103] (3) Flow cytometry was used to detect the fluorescence intensity of the fluorescent markers in the cell suspension. The fluorescence intensity of Eu / Ce DPA@TPP-DOX@HA was detected using the ECD channel.

[0104] from Figure 12 It can be seen that the fluorescence intensity of the material in the normal thyroid cells NTHY-ORI-3-1 is significantly lower than that in the anaplastic thyroid cancer cells CAL-62, indicating that the anaplastic thyroid cancer cells CAL-62 have a strong uptake ability for Eu / CeDPA@TPP-DOX@HA.

[0105] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.

Claims

1. A method for preparing a lanthanide metal-organic framework composite material, characterized in that: The following steps are involved: The lanthanide metal salt and the organic ligand are mixed and reacted in the form of a solution to obtain an organic framework material; The triphenylphosphine-modified doxorubicin and the organic framework material are mixed and reacted in the form of a solution to obtain an organic framework material carrying TPP-DOX; The polysaccharide having the function of targeting the CD44 receptor on the surface of cancer cells and the organic framework material carrying TPP-DOX are mixed and reacted in the form of a solution to obtain the lanthanide metal-organic framework composite material; The lanthanide metal salts include cerium nitrate and europium nitrate; The organic ligand includes pyridine-2,6-dicarboxylic acid; The polysaccharide having the function of targeting CD44 receptor on the surface of cancer cells includes hyaluronic acid.

2. The preparation method according to claim 1, characterized in that The molar ratio of the cerium nitrate, europium nitrate and pyridine-2,6-dicarboxylic acid is 0.125:0.125:(0.1-0.5).

3. The preparation method according to claim 1, characterized in that The lanthanide metal salt is mixed with the organic ligand twice.

4. The preparation method according to claim 1, characterized in that The mass ratio of the organic framework material to triphenylphosphine-modified doxorubicin is 20:

4.

5. The preparation method according to claim 1, characterized in that The time for mixing the triphenylphosphine-modified doxorubicin and the organic framework material in the form of a solution to react is 6 hours.

6. The preparation method according to claim 1, characterized in that The mass ratio of the organic skeleton material carrying TPP-DOX to the polysaccharide having the function of targeting the CD44 receptor on the surface of cancer cells is 1:

1.

7. The preparation method according to claim 1, characterized in that The polysaccharide having the function of targeting the CD44 receptor on the surface of cancer cells and the organic framework material carrying TPP-DOX are mixed in the form of a solution and reacted for 24 hours.

8. A lanthanide metal-organic framework composite material prepared by the preparation method according to any one of claims 1 to 7.

9. Use of the lanthanide metal-organic framework composite material according to claim 8 in preparing drugs for targeted tumor treatment.

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