Lanthanide metal organic framework composite material as well as preparation method and application thereof

Abstract

The invention discloses a lanthanide series metal organic framework composite material as well as a preparation method and application thereof, and belongs to the technical field of preparation of targeting materials. The preparation method of the lanthanide series metal organic framework composite material comprises the following steps: mixing lanthanide series metal salt and an organic ligand in a solution form for reaction to obtain an organic framework material; the preparation method comprises the following steps: mixing triphenylphosphine modified adriamycin and an organic framework material in a solution form for reaction to obtain a TPP-DOX loaded organic framework material; polysaccharide with a cancer cell surface CD44 receptor targeting function and an organic framework material carrying TPP-DOX are mixed and react in a solution form, and the lanthanide series metal organic framework composite material is obtained. The lanthanide series metal organic framework composite material prepared by the invention can accurately identify and position tumor tissues, and does not generate adverse effects on surrounding normal tissues.

Description

Technical Field

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

[0002] Once most cancers are diagnosed, they are basically in the middle or late stages, which means that the best period for surgical treatment has been missed. Early diagnosis and early treatment are crucial. Traditional cancer treatment methods include surgery, radiotherapy, and chemotherapy, but these methods are largely restricted by targeting issues. Therefore, actively developing new targeted precision treatment methods is a difficult and hot topic in current research.

[0003] Recent studies have involved a variety of new materials for the treatment of malignant tumors, including metal organic frameworks, biodegradable polymers, nanoparticles, nanomagnetic materials, etc. The research and application of these materials provide new ideas and methods for tumor treatment, and are expected to bring more effective and safer treatment strategies for clinical tumor treatment. 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, neodymium, etc.) and organic ligands (such as carboxylic acids, pyridine, etc.) through coordination 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 characteristics can be regulated 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 new type of material, Ln-MOFs have shown potential application value in the treatment of tumors, including drug delivery, photothermal therapy, photodynamic therapy, tumor imaging, etc., but its biocompatibility and toxicity in the body remain a key issue. Some Ln-MOFs may cause immune responses or toxic effects, limiting their clinical application. Although Ln-MOFs can achieve targeted delivery to tumors through functionalization or modification, their targeting performance needs to be further improved. How to ensure that Ln-MOFs can accurately identify and locate tumor tissues 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: 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] Further, 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 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 for a reaction time of 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 by combining pyridine-2,6-dicarboxylic acid (DPA) with metal ions cerium and europium (Eu / Ce) in the present invention is a special type of bimetallic lanthanide organic framework. As a new type of material, it shows potential application value in drug delivery and tumor imaging in the treatment of tumors. Triphenylphosphine-Doxorubicin (TPP-DOX) modified with triphenylphosphine is a drug delivery system with positive charge and the function of targeting negatively charged mitochondria; hyaluronic acid (HA) is a polysaccharide substance with the function of targeting CD44 receptors on the surface of cancer cells. The new lanthanide metal organic framework composite material (Eu / Ce DPA@TPP-DOX@HA) synthesized by using Eu / Ce DPA, TPP-DOX and HA has multiple functions such as targeted delivery, drug protection and controlled release, optical labeling, good biocompatibility and small side effects.

[0021] Eu / Ce DPA is a special type of lanthanide metal organic framework, which contains two different metal ions (Eu / Ce) in its structure. This bimetallic structure can introduce more active sites to MOF or adjust its surface chemical properties, thereby improving its performance in gas adsorption, catalysis, separation and other fields. Eu / Ce DPA was used as the skeleton of the target material, and TPP-DOX was loaded to synthesize Eu / Ce DPA@TPP-DOX, which improved the in vivo stability and tumor targeting of Eu / Ce DPA@TPP-DOX. HA was connected to the surface of Eu / Ce-DPA@TPP-DOX, which improved the imaging effect and targeting effect of the tumor, forming the final tumor-killing composite material with targeted sensitivity and specificity, namely 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 treatment of tumors.

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

[0025] The lanthanide metal organic framework composite material prepared by the invention can accurately identify and locate tumor tissues and will not have 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 drawings required for use in the embodiments will be briefly introduced below. 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 creative work.

[0028] Figure 1 The characterization results of Eu / Ce DPA prepared in Example 1, wherein a is a TEM image and b is an EDS analysis result;

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

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

[0031] Figure 4 This is a high-resolution spectrum of Eu / Ce DPA@TPP-DOX@HA prepared in Example 1, wherein 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 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, raw material TPP-DOX used in Example 1, raw material HA used 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] Fig. 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] Fig.10 The expression of CD44 in normal thyroid cells NTHY-ORI-3-1 and anaplastic thyroid cancer CAL-62 cells;

[0038] Fig.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), wherein a is Eu / CeDPA, b is Eu / Ce DPA@TPP-DOX, and c is Eu / Ce DPA@TPP-DOX@HA;

[0039] Fig.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 should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

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

[0042] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation 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 a conflict with any incorporated document, the content 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 of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are 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] (I) Synthesis of Eu / Ce DPA (organic framework material):

[0048] (1) Dissolve 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) 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) Take 100 μL of the above metal ion solution and add it dropwise to the organic ligand solution, and ultrasonically react for 10 min (ultrasonic power is 40 W). Then, the remaining metal ion solution is added dropwise to the mixed solution and ultrasonically reacted for 60 min. The reaction system will gradually produce a yellow precipitate, which is centrifuged (10,000 rpm, 3 min), washed twice with ethanol, and freeze-dried overnight to obtain Eu / CeDPA, a yellow solid powder.

[0051] (II) 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 and transparent 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), ethanol washing twice, and freeze-drying overnight, Eu / Ce DPA@TPP-DOX (organic framework material carrying TPP-DOX) was obtained, which was a purple solid powder.

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

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

[0057] Hyaluronic acid (HA, 20 mg) was dissolved in 1 mL of primary 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), ethanol washing twice, and freeze-drying 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 presents a flaky nanostructure, and its EDS results show that 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. 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 not only contains Eu, Ce and N elements in Eu / Ce DPA, but also contains P element unique to TPP-DOX. The elements are evenly distributed with an average size of about 250nm.

[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 the DPA carbonyl group is 1704.8 cm -1 The synthesized Eu / Ce DPA red-shifts 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: C=O (288.1eV), CH (285.2eV), and CC (284.4eV). The Eu 3d spectrum ( Figure 4 f) shows that there are two characteristic peaks, Eu 3d 5 / 2 (1134.9 eV) and Eu 3d 3 / 2 (1164.6 eV), and there is a characteristic satellite peak of Eu 3d (1143.1 eV) between the two diffraction peaks. After loading TPP-DOX, the unique characteristic peak of the P element appears at 190.4 eV. 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 content was 18.35 ppm (mg / g) and the Ce content was 18.11 ppm (mg / g). The ratio of the two was about 1, which was consistent with the Eu content during synthesis. 3+ / Ce 3+ The feed ratio is consistent. In Eu / Ce DPA@TPP-DOX@HA (100 μg / mL), the Eu element content is 6.95 ppm (mg / g), and the Ce element content is 5.49 ppm (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 Figure 6 .

[0071] Zeta potential diagrams of Eu / Ce DPA (MOF) prepared in Example 1, raw material TPP-DOX used in Example 1, raw material HA used 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 Figure 7 .

[0072] In Tris-HCl buffer solution (10mM, pH 8.5), negatively charged Eu / Ce DPA and positively charged TPP-DOX synthesized positively charged Eu / Ce DPA@TPP-DOX through electrostatic interaction, and negatively charged HA was also coated on the surface of Eu / Ce DPA@TPP-DOX through electrostatic interaction. The successful loading of TPP-DOX and the successful coating of HA were verified by UV absorption spectroscopy. After loading TPP-DOX and coating HA, it can be seen that the UV absorption spectrum is significantly improved.

[0073] (6) Fluorescence emission spectra of Eu / Ce DPA@TPP-DOX@HA prepared in Example 1 under different excitation lights. 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 Fig. 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 ultraviolet 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 mixture was reacted on a constant temperature oscillator 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 marker (MOF+TMB group). The control group was a group without the addition of Eu / Ce DPA, and the ultraviolet absorption spectrum of the reaction system was measured using an enzyme marker (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), and the reaction was carried out on a constant temperature oscillator at a shaking rate of 300 rpm and a reaction condition of 37°C for 30 min. The ultraviolet absorption spectrum of the reaction system was measured using an enzyme marker (MOF+TMB+H2O2 group). The control group was not added with Eu / Ce DPA, and the ultraviolet absorption spectrum of the reaction system was measured using an enzyme marker (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), and the mixture was reacted at 37°C on a constant temperature shaker at a shaking rate of 300 rpm for 30 min. The UV absorption spectrum of the reaction system was measured using an ELISA instrument (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 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 UV absorption spectrum of the reaction system was measured using an enzyme marker (MTH+TMB+H2O2 group).

[0080] Effect Example 2

[0081] In order 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. Fig.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 the material solution to the cells was 200 μL:1.5 mL) to obtain a cell suspension;

[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 mixture of cells and antibodies 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 Fig.10 It can be seen that the expression level of CD44 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 experiment verified the killing ability of Eu / Ce DPA, Eu / Ce DPA@TPP-DOX and Eu / Ce DPA@TPP-DOX@HA prepared in Example 1 against cancer cells (CAL-62 cells) and normal tissue cells (NTHY-ORI-3-1 cells) at different concentrations. Fig.11 . Fig.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), and 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 a 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 the thyroid normal cells NTHY-ORI-3-1 and thyroid anaplastic cancer cells CAL-62 with Eu / CeDPA@TPP-DOX@HA solution for 6 hours, the cell fluorescence intensity was detected by flow cytometry. The results are shown in Fig.12 .

[0100] The specific verification method is as follows:

[0101] (1) Cultivate NTHY-ORI-3-1 and CAL-62 cells 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, and the material solutions of different concentrations were mixed with cells (the volume ratio of the material solution to the 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 Fig.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 only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined 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.

2. The preparation method according to claim 1, characterized in that: The lanthanide metal salts include cerium nitrate and europium nitrate; The organic ligand includes pyridine-2,6-dicarboxylic acid; 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 for a reaction time of 24 hours; The polysaccharide having the function of targeting CD44 receptor on the surface of cancer cells comprises hyaluronic acid.

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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