A nano-drug for treating rheumatoid arthritis and a preparation method thereof

Abstract

The application discloses a kind of nanomedicine for treating rheumatoid arthritis, including triptolide, Y6 fluorescent probe and poloxamer, triptolide and Y6 fluorescent probe are polymerized by poloxamer to form triptolide nanomedicine Y6@TPL, the average particle size of the triptolide nanomedicine Y6@TPL is 20~300nm.The application combines triptolide with Y6 fluorescent probe, constructs a new type of nano delivery system, the system not only realizes the accurate control release of drug, but also can be enhanced by photo-thermal response Targeted accumulation of rheumatoid arthritis lesion site, significantly improve the selective killing effect of triptolide on inflammatory synoviocytes, while greatly reducing the toxic side effects on normal joint tissue.The nano system integrates photo-thermal therapy, photo-thermal imaging and targeted drug delivery functions, and provides an integrated solution for the diagnosis and treatment of RA.

Description

Technical Field

[0001] This invention belongs to the field of nanomedicine technology, specifically relating to a nanomedicine for treating rheumatoid arthritis and its preparation method. Background Technology

[0002] Triptolide (TPL) is a key active ingredient isolated and extracted from the traditional Chinese medicine Tripterygium wilfordii Hook.f. It exhibits strong anti-inflammatory, anti-tumor, and immunosuppressive pharmacological effects, showing great potential in the treatment of autoimmune diseases and malignant tumors. However, although the epoxy groups in its structure endow it with activity, they also result in poor water solubility, low bioavailability, short half-life, and organ toxicity such as liver and kidney toxicity, which limits its further clinical application.

[0003] Current development of triptolide drug carriers (such as liposomes, nanoparticles, and hydrogels) mainly focuses on addressing its application bottlenecks in the treatment of rheumatoid arthritis: improving the solubility of hydrophobic drugs, enhancing joint lesion targeting, and reducing systemic toxicity to broaden its therapeutic window for diseases like rheumatoid arthritis. For example, patent CN118831177A discloses a triptolide liposome prepared using a thin-film dispersion method. However, existing carrier systems still suffer from complex synthesis processes, making it difficult to meet the large-scale requirements for long-term use of chronic rheumatoid arthritis medication, insufficient joint targeting efficiency, and unclear metabolic mechanisms of carrier materials in synovial tissue. Breakthroughs in these key technologies are crucial for promoting the clinical application of triptolide in the treatment of rheumatoid arthritis. Summary of the Invention

[0004] Based on the shortcomings of the existing technology, the purpose of this invention is to develop a nanomedicine formed by assembling triptolide and Y6 fluorescent probe and its preparation method, and to use photothermal therapy to synergistically treat rheumatoid arthritis.

[0005] The technical solution adopted by the present invention to solve the above-mentioned technical problem is as follows:

[0006] The present invention discloses a nanomedicine for treating rheumatoid arthritis, comprising triptolide, Y6 fluorescent probe and poloxamer, wherein triptolide and Y6 fluorescent probe are assembled from poloxamer polymer to form the nanomedicine, and the average particle size of the nanomedicine is 20-300 nm.

[0007] Preferably, the mass ratio of the Y6 fluorescent probe to poloxamer is 1:10 to 1:2, and the mass of triptolide does not exceed 10% of the mass of poloxamer.

[0008] A method for preparing a nanomedicine for treating rheumatoid arthritis includes the following steps:

[0009] Step S01: Material preparation. Determine the amount of each component, weigh or measure the materials and set them aside for later use. The materials include triptolide, Y6 fluorescent probe, poloxamer, organic solvent and poor solvent for triptolide.

[0010] Step S02: Dissolve triptolide, Y6 fluorescent probe and poloxamer thoroughly in an organic solvent to obtain solution A;

[0011] Step S03: Place the poor solvent of triptolide in a glass bottle, add a magnetic stir bar, place it on a magnetic stirrer and stir, and under stirring conditions, add solution A dropwise to the poor solvent of triptolide to obtain mixture B;

[0012] Step S04: Stir the mixture B until the organic solution has completely evaporated to obtain solution C;

[0013] Step S05: Centrifuge solution C, discard the supernatant, and remove the poor solvent of triptolide, unencapsulated drug and fluorescent probe to obtain the substrate;

[0014] Step S06: Add an equal amount of triptolide poor solvent as in S03 to the substrate to obtain the finished triptolide nanomedicine Y6@TPL.

[0015] Preferably, in step S01, triptolide is a Tripterygium wilfordii extract with a purity of over 90%, the organic solvent is tetrahydrofuran, and the unsuitable solvent for triptolide is double-distilled water.

[0016] Preferably, in step S02, the concentration of poloxamer in the organic solvent is 5~30 mg / ml.

[0017] Preferably, in step S03, the volume ratio of solution A to triptolide, a poor solvent, is 1:5.

[0018] Preferably, in step S03, the fixed rotation speed of the magnetic stirrer is 1500 rpm / min.

[0019] Preferably, in step S02, an RITC fluorescent probe is added to solution A, wherein the mass of the RITC fluorescent probe is 3% to 5% of the mass of the Y6 fluorescent probe.

[0020] Preferably, the triptolide nanoparticle Y6@TPL has a particle size PDI of 0.3~0.5, a zeta potential of -6mV~-35mV, and a size range of 24~122nm.

[0021] Preferably, the triptolide nanomedicine Y6@TPL is an injectable formulation, used in conjunction with photothermal therapy for synergistic treatment.

[0022] Preferably, the drug is an injectable formulation that can be used in conjunction with photothermal synergistic therapy.

[0023] Compared with the prior art, the advantages of the present invention are:

[0024] 1. This invention innovatively combines triptolide (TPL) with a Y6 fluorescent probe to construct a novel nanodelivery system. This system not only achieves precise controlled drug release but also enhances targeted accumulation at rheumatoid arthritis lesions through photothermal response, significantly improving the selective killing effect of TPL on inflamed synovial cells while greatly reducing toxic side effects on normal joint tissues. This nanosystem integrates photothermal therapy, photothermal imaging, and targeted drug delivery functions, providing an integrated solution for the diagnosis and treatment of RA.

[0025] 2. In terms of photothermal controlled release technology, a novel fluorescent probe Y6 has been introduced. Compared with traditional fluorescent probes, it has excellent photothermal stability and biocompatibility, and exhibits lower spontaneous background fluorescence, deeper tissue penetration, and higher signal-to-background ratio during imaging.

[0026] 3. By using near-infrared light (808nm) irradiation to trigger TPL release, precise controlled release of the drug is achieved, which not only improves the therapeutic effect but also reduces the side effects caused by premature drug release.

[0027] 4. In terms of synergistic treatment strategies, combining photothermal therapy (PTT) and drug therapy with TPL, the photothermal effect converts light energy into heat energy under external near-infrared light irradiation, directly inhibiting inflammatory cells. Local heat therapy at the joints enhances membrane permeability, enhances the cells' ability to absorb antirheumatic drugs, and activates drug-sensitive inflammatory cells, achieving better anti-inflammatory effects at low drug doses. Attached Figure Description

[0028] Figure 1 The molecular structures of TPL, Y6, RITC and poloxamer involved in this invention are shown in the diagram.

[0029] Figure 2 The DLS particle size distribution diagrams of the triptolide nanomedicine Y6@TPL of Examples 1 and 2 of this invention are shown.

[0030] Figure 3 This is a TEM image of the triptolide nanomedicine Y6@TPL of Example 3 of the present invention;

[0031] Figure 4 The particle size distribution diagram of the triptolide nanomedicine Y6@TPLDLS of Example 3 of the present invention is shown.

[0032] Figure 5 The Zeta potential diagram of the triptolide nanomedicine Y6@TPL in Example 3 of this invention;

[0033] Figure 6 The ultraviolet spectrum of the triptolide nanomedicine Y6@TPL of Example 3 of the present invention;

[0034] Figure 7 The fluorescence spectrum of the triptolide nanomedicine Y6@TPL in Example 3 of this invention;

[0035] Figure 8 Images of RAW264.7 macrophages uptake of triptolide nanoparticles Y6@TPL at different time points in Example 4 of this invention;

[0036] Figure 9 Cytotoxicity diagram of the triptolide nanomedicine Y6@TPL in RAW264.7 macrophage cck8 cells, as described in Example 4 of this invention;

[0037] Figure 10 The figure shows the temperature rise of the triptolide nanomedicine Y6@TPL under 808nm irradiation in Example 3 of the present invention.

[0038] Figure 11 This is a graph showing the temperature rise of the triptolide nanomedicine Y6@TPL at different concentrations in Example 3 of the present invention.

[0039] Figure 12 Photothermal stability diagram of the triptolide nanomedicine Y6@TPL in Example 3 of this invention;

[0040] Figure 13 This is an L fluorescence imaging image of the triptolide nanomedicine Y6@TPL at different concentrations in Example 3 of the present invention. Detailed Implementation

[0041] The technical solution of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0042] This invention discloses a nanomedicine for treating rheumatoid arthritis, comprising triptolide, a Y6 fluorescent probe, and poloxamer. Triptolide and the Y6 fluorescent probe are polymerized with poloxamer to form triptolide nanomedicine Y6@TPL. The average particle size of triptolide nanomedicine Y6@TPL is 20-300 nm, the particle size distribution (PDI) is 0.3-0.5, the zeta potential is -6 mV to -35 mV, and the size range is 24-122 nm. Triptolide nanomedicine Y6@TPL is an injectable formulation, used in conjunction with photothermal therapy for synergistic treatment.

[0043] A method for preparing a nanomedicine for treating rheumatoid arthritis includes the following steps:

[0044] Step S01: Material preparation. Determine the amount of each component, weigh or measure the materials and set them aside for later use. The materials include triptolide, Y6 fluorescent probe, poloxamer, organic solvent, and a poor solvent for triptolide. Among them, triptolide is an extract of Tripterygium wilfordii with a purity of over 90% and its source is not limited. The organic solvent is tetrahydrofuran, and the poor solvent for triptolide is double-distilled water.

[0045] Step S02: Place the organic solvent in a beaker, and fully dissolve triptolide, Y6 fluorescent probe, and poloxamer in the organic solvent to obtain solution A. The volume of solution A remains essentially unchanged compared to the organic solvent. The concentration of poloxamer in the organic solvent is 5-30 mg / ml, the mass ratio of Y6 fluorescent probe to poloxamer is 1:10-1:2, and the mass of triptolide does not exceed 10% of the mass of poloxamer.

[0046] Step S03: Place the poor solvent of triptolide in a glass bottle, add a magnetic stir bar to the poor solvent of triptolide, place the glass bottle on a magnetic stirrer and stir, and under stirring conditions, add solution A dropwise to the poor solvent of triptolide to obtain mixture B; wherein, the fixed rotation speed of the magnetic stirrer is 1500 rpm / min, and the volume ratio of solution A to poor solvent of triptolide is 1:5;

[0047] Step S04: Stir the mixture B until the organic solution has completely evaporated to obtain solution C;

[0048] Step S05: Centrifuge solution C and discard the supernatant to remove the poor solvent of triptolide, unencapsulated drug and fluorescent probe, and obtain the substrate;

[0049] Step S06: Add an equal amount of triptolide poor solvent as in S03 to the substrate to obtain the finished triptolide nanomedicine Y6@TPL.

[0050] In another embodiment, an RITC fluorescent probe is added to solution A obtained in step S02, wherein the mass of the RITC fluorescent probe is 3% to 5% of the mass of the Y6 fluorescent probe, to achieve visualization of the triptolide nanoparticle Y6@TPL in the visible light region. Laser confocal imaging is used to observe the acquisition images of the triptolide nanoparticle Y6@TPL in RAW264.7 macrophages over time.

[0051] The cytotoxicity of the triptolide nanomedicine Y6@TPL was evaluated using the RAW264.7 macrophage model, and the non-toxic dose range of its active ingredient, triptolide, was finally determined.

[0052] Tripterygium wilfordii nanoparticles Y6@TPL can be used in drugs for the treatment of rheumatoid arthritis. There are no particular restrictions on pharmaceutically acceptable excipients, as long as they do not damage the nanoparticle structure of triptolide nanoparticles Y6@TPL.

[0053] Y6 (also known as BTP-4F), CAS2304444-49-1, is a highly conjugated electron-deficient organic semiconductor with an A-DAD-A structure. This molecule consists of a fused-ring thienothiophene-indole (TTP-TTI) core group and a 2-(5,6-difluoro-3-oxo-2,3-dihydro-1H-inden-1-yl)malononitrile (2FIC) end group. The 2FIC end group is believed to promote intermolecular interactions and enhance light absorption. The absorption spectrum of Y6 peaks at 810 nm and extends to 1100 nm, indicating that Y6 and its polymer blends possess the potential to cover the entire visible to near-infrared spectrum.

[0054] RITC (also known as Rhodamine B isothiocyanate), CAS 944130-99-8, is a cell-permeable fluorescent dye, most commonly used as a conjugate for antibodies and proteins for fluorescence detection in microscopy. Its excitation and emission spectra peak wavelengths are 570 nm and 595 nm, respectively, and it can be combined with immunohistochemistry for labeling studies. Y6 combined with RITC labeling can be used for fluorescent labeling studies of nanomedicines in the visible light range.

[0055] like Figure 1 The molecular structures of TPL, Y6, RITC, and poloxamer are shown.

[0056] The present invention includes the following embodiments:

[0057] Table 1 lists the raw materials and their contents involved in Examples 1-4:

[0058]

[0059] The preparation methods of Examples 1-4 are as follows:

[0060] Example 1:

[0061] Take 1.5 mg of triptolide, 2 mg of Y6 fluorescent probe, and 20 mg of poloxamer, and dissolve them in 2 ml of tetrahydrofuran organic solution to obtain solution A1. Add 10 ml of double-distilled water to a magnetic stir bar and place it on a magnetic stirrer. Stir at 1500 rpm / min. Under stirring conditions, add solution A1 dropwise to the double-distilled water to obtain mixture B1. Stir mixture B1 and evaporate for 3 days until the organic solution is completely evaporated to obtain solution C1. Centrifuge solution C1 and discard the supernatant to obtain substrate 1. Add 10 ml of double-distilled water to substrate 1 to obtain triptolide nanomedicine Y6@TPL.

[0062] Example 2:

[0063] Take 1 mg of triptolide, 10 mg of Y6 fluorescent probe, and 20 mg of poloxamer, and dissolve them in 2 ml of tetrahydrofuran organic solution to obtain solution A2. Add 10 ml of double-distilled water to a magnetic stir bar and place it on a magnetic stirrer. Stir at 1500 rpm / min. Under stirring conditions, add solution A2 dropwise to the double-distilled water to obtain mixture B2. Stir mixture B2 and evaporate for 3 days until the organic solution is completely evaporated to obtain solution C2. Centrifuge solution C2 and discard the supernatant to obtain substrate 2. Add 10 ml of double-distilled water to substrate 2 to obtain triptolide nanomedicine Y6@TPL.

[0064] Example 3:

[0065] Take 0.2 mg of triptolide, 5 mg of Y6 fluorescent probe, and 20 mg of poloxamer, and dissolve them in 2 ml of tetrahydrofuran organic solution to obtain solution A3. Add 10 ml of double-distilled water to a magnetic stir bar and place it on a magnetic stirrer. Stir at 1500 rpm / min. Under stirring conditions, add solution A3 dropwise to the double-distilled water to obtain mixture B3. Stir mixture B3 and evaporate for 3 days until the organic solution is completely evaporated to obtain solution C3. Centrifuge solution C3 and discard the supernatant to obtain substrate 3. Add 10 ml of double-distilled water to substrate 3 to obtain triptolide nanomedicine Y6@TPL.

[0066] Example 4:

[0067] Take 0.2 mg of triptolide, 5 mg of Y6 fluorescent probe, 20 mg of poloxamer, and 0.2 mg of RITC fluorescent probe, and dissolve them in 2 ml of tetrahydrofuran organic solution to obtain solution A4. Add 10 ml of double-distilled water to a magnetic stir bar and place it on a magnetic stirrer. Stir at 1500 rpm / min. Under stirring conditions, add solution A4 dropwise to the double-distilled water to obtain mixture B4. Stir mixture B4 and evaporate for 3 days until the organic solution is completely evaporated to obtain solution C4. Centrifuge solution C4 and discard the supernatant to obtain substrate 4. Add 10 ml of double-distilled water to substrate 4 to obtain triptolide nanomedicine Y6@TPL.

[0068] In the above Examples 1-4, Examples 1-3 are the particle size optimization process of triptolide nanomedicine Y6@TPL, Example 3 is the triptolide nanomedicine Y6@TPL with optimized particle size, and Examples 1-3 do not add RITC fluorescent probe, while Example 4 adds RITC fluorescent probe based on Example 3.

[0069] Performance tests were conducted on the above Examples 1-4. Specifically, the particle size of the triptolide nanoparticles Y6@TPL in Examples 1-2 was measured, and the performance of the triptolide nanoparticles Y6@TPL in Example 3 was tested, including TEM testing, DLS particle size distribution testing, Zeta potential testing, ultraviolet spectroscopy testing, and fluorescence spectroscopy testing. The triptolide nanoparticles Y6@TPL in Example 4 were subjected to macrophage cell uptake experiments, and cytotoxicity was verified.

[0070] Combination Figure 2-9 The specific test results are as follows:

[0071] (1) such as Figure 2 A and Figure 2 As shown in B, the size of the triptolide nanoparticle Y6@TPL in Example 1 is 21~58nm, and the size of the triptolide nanoparticle Y6@TPL in Example 2 is 190~295nm. The triptolide nanoparticle Y6@TPL prepared in Examples 1 and 2 are uniform in size and uniformly dispersed.

[0072] (2) Figure 3 This is a TEM image of the triptolide nanomedicine Y6@TPL from Example 3. Figure 4 This is the DLS particle size distribution diagram of the triptolide nanomedicine Y6@TPL in Example 3, from... Figure 3 and Figure 4 It can be seen that the triptolide nanoparticles Y6@TPL prepared in Example 3 have uniform size and dispersion, with a size of 24-122 nm. Figure 5This is the Zeta potential diagram of the triptolide nanomedicine Y6@TPL in Example 3, with a Zeta potential of -22mV. Figure 6 The UV spectrum of the triptolide nanomedicine Y6@TPL in Example 3 shows that the material has a characteristic absorption peak at 470 nm, which can be used for bioimaging. Figure 7 The fluorescence spectrum of the triptolide nanomedicine Y6@TPL in Example 3 shows that the emission spectrum peaks at 945 nm and extends to 1200 nm. This means that the triptolide nanomedicine Y6@TPL exhibits significant fluorescence emission in the NIR-II region and has the potential for deep tissue and high-resolution bioimaging.

[0073] (3) The steps and results of the RAW264.7 macrophage cell uptake experiment in Example 4 are as follows:

[0074] RAW264.7 macrophages were seeded onto 35 mm laser confocal microplates. 1 ml of triptolide nanoparticles (Y6@TPL) and 1 ml of complete culture medium were incubated with the cells for 0 h, 1 h, and 2 h, respectively. The culture medium was then removed, followed by washing with PBS, fixation with paraformaldehyde solution, quenching with glycine solution, permeabilization, washing with PBS, staining with DAPI in the dark, washing with PBS, and then performing laser confocal microscopy. The results are shown below. Figure 8 As shown, the triptolide nanomedicine Y6@TPL can be internalized into RAW264.7 macrophages within 1 hour, and the signal of the triptolide nanomedicine Y6@TPL is enhanced over time.

[0075] (4) The steps and results of the nanomedicine cytotoxicity test in Example 4 are as follows:

[0076] RAW264.7 macrophages were seeded at 2 × 10⁵ cells / well in 96-well plates, with each well containing 1 μg / mL LPS for induction. After 24 h of culture, the old culture medium was discarded, and DMEM medium containing triptolide nanoparticles Y6@TPL at concentrations of 2 ng / ml, 4 ng / ml, 8 ng / ml, 10 ng / ml, 12 ng / ml, 14 ng / ml, 16 ng / ml, and 32 ng / ml was added sequentially (n=6) to form the experimental group. A control group was also set up, which contained only DMEM and no triptolide nanoparticles Y6@TPL. After culturing both groups for 24 h, CCK-8 reagent was added, and after 4 h of incubation, the absorbance was measured at 490 nm. Cell viability was calculated based on the absorbance ratio of the experimental group to the control group. Figure 9As shown, the results indicated that when the concentration of the triptolide nanoparticle Y6@TPL was 4 ng / ml, the cell survival rate was 96.3%; while when the concentration of the triptolide nanoparticle Y6@TPL was 8.448 ng / ml, the cell survival rate was 50%. This suggests that triptolide at a concentration of 8.448 ng / ml exhibited cytotoxicity, therefore, when the concentration of triptolide was less than 4 ng / ml, the cytotoxicity was negative.

[0077] like Figure 10-13 As shown in Example 3, the present invention also provides a method for photothermal evaluation of the triptolide nanomedicine Y6@TPL, comprising the following steps:

[0078] S1. Evaluation of photothermal effect:

[0079] Based on the mass concentration of the Y6 fluorescent probe, the triptolide nanoparticles Y6@TPL were diluted to gradient concentrations of 10, 20, 40, and 80 μg / ml. The temperature rise of the triptolide nanoparticles Y6@TPL was observed under 808 nm laser irradiation at a distance of 2 cm from the sample. The evaluation results are detailed below. Figure 10-11 As shown, within a certain range, the higher the concentration of the triptolide nanomedicine Y6@TPL, the faster the heating rate and the higher the final temperature.

[0080] S2. Evaluation of photothermal stability:

[0081] Y6 fluorescent probe was taken as 80 μg / ml of triptolide nanoparticle Y6@TPL. The sample was irradiated with an 808 nm laser at a distance of 2 cm from the Y6@TPL sample for 5 min, followed by 5 min of cooling. This process was repeated 4 times, and the sample temperature rise and cooldown were measured each time. The evaluation results are as follows: Figure 12 As shown, after four consecutive laser irradiation cycles, the temperature of the triptolide nanoparticle Y6@TPL solution can still reach 47℃, indicating that the triptolide nanoparticle Y6@TPL has good photothermal stability under laser irradiation.

[0082] S3. Fluorescence Imaging Evaluation:

[0083] Tripterygium wilfordii nanoparticles Y6@TPL with Y6 fluorescent probe concentrations of 0, 10, 20, and 40 μg / ml were subjected to near-infrared II fluorescence imaging; the evaluation results are as follows. Figure 13 As shown, the fluorescence signal is stronger with the increase of the concentration of the triptolide nanomedicine Y6@TPL.

[0084] This invention focuses on innovative research into the treatment of rheumatoid arthritis (RA). First, a novel nanomedicine (triptolide nanomedicine Y6@TPL) was designed and synthesized. By combining the polarity of the polymer with the Y6 fluorescent probe, controlled drug release and photothermal effects were achieved, significantly improving drug selectivity and reducing the toxicity of TPL to normal cells. Regarding the photothermal controlled release technology, the novel fluorescent probe Y6 was introduced. Compared to traditional fluorescent probes, it exhibits excellent photothermal stability and biocompatibility, with lower spontaneous background fluorescence, deeper tissue penetration, and a higher signal-to-background ratio during imaging. Near-infrared light (808nm) irradiation was used to trigger TPL release, achieving precise controlled drug release, which improved therapeutic efficacy and reduced the side effects of premature drug release. In terms of synergistic treatment strategies, photothermal therapy (PTT) and TPL drug therapy were combined. The photothermal effect converts light energy into heat energy under external near-infrared light irradiation, directly inhibiting inflammatory cells. Local heat therapy at the joint enhances membrane permeability, strengthens the cells' ability to absorb anti-rheumatic drugs, and activates drug-sensitive inflammatory cells, achieving better anti-inflammatory effects at low drug doses. Ultimately, this nanosystem integrates photothermal therapy, photothermal imaging, and targeted drug delivery, providing an all-in-one solution for the diagnosis and treatment of RA.

[0085] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A nanomedicine for treating rheumatoid arthritis, characterized in that: The nanoparticles Y6@TPL are composed of triptolide, a Y6 fluorescent probe, and poloxamer. Triptolide and the Y6 fluorescent probe are polymerized with poloxamer to form triptolide nanoparticles Y6@TPL. The average particle size of the triptolide nanoparticles Y6@TPL is 20-300 nm. The molecular structure of Y6 is as follows: .

2. The nanomedicine for treating rheumatoid arthritis according to claim 1, characterized in that: The mass ratio of the Y6 fluorescent probe to poloxamer is 1:10 to 1:2, and the mass of triptolide does not exceed 10% of the mass of poloxamer.

3. A method for preparing a nanomedicine for treating rheumatoid arthritis according to any one of claims 1-2, characterized in that, Includes the following steps: Step S01: Material preparation. Determine the amount of each component, weigh or measure the materials and set them aside for later use. The materials include triptolide, Y6 fluorescent probe, poloxamer, organic solvent and poor solvent for triptolide. Step S02: Dissolve triptolide, Y6 fluorescent probe and poloxamer thoroughly in an organic solvent to obtain solution A; Step S03: Place the poor solvent of triptolide in a glass bottle, add a magnetic stir bar, place it on a magnetic stirrer and stir, and under stirring conditions, add solution A dropwise to the poor solvent of triptolide to obtain mixture B; Step S04: Stir the mixture B until the organic solvent has completely evaporated to obtain solution C; Step S05: Centrifuge solution C, discard the supernatant, and remove the poor solvent of triptolide, unencapsulated drug and fluorescent probe to obtain the substrate; Step S06: Add an equal amount of triptolide poor solvent as in S03 to the substrate to obtain the finished triptolide nanomedicine Y6@TPL.

4. The method for preparing a nanomedicine for treating rheumatoid arthritis according to claim 3, characterized in that: In step S01, triptolide is a Tripterygium wilfordii extract with a purity of over 90%, the organic solvent is tetrahydrofuran, and the unsuitable solvent for triptolide is double-distilled water.

5. The method for preparing a nanomedicine for treating rheumatoid arthritis according to claim 3, characterized in that: In step S02, the concentration of poloxamer in the organic solvent is 5~30 mg / ml.

6. The method for preparing a nanomedicine for treating rheumatoid arthritis according to claim 3, characterized in that: In step S03, the volume ratio of solution A to triptolide, a poor solvent, is 1:

5.

7. The method for preparing a nanomedicine for treating rheumatoid arthritis according to claim 3, characterized in that: In step S03, the fixed rotation speed of the magnetic stirrer is 1500 rpm / min.

8. The method for preparing a nanomedicine for treating rheumatoid arthritis according to claim 3, characterized in that: In step S02, an RITC fluorescent probe is added to solution A, wherein the mass of the RITC fluorescent probe is 3% to 5% of the mass of the Y6 fluorescent probe.

9. The method for preparing a nanomedicine for treating rheumatoid arthritis according to claim 3, characterized in that: The triptolide nanoparticles Y6@TPL have a particle size distribution (PDI) of 0.3–0.5 mm, a zeta potential of -6 mV to -35 mV, and a size range of 24–122 nm.

10. The application of the triptolide nanomedicine Y6@TPL as described in any one of claims 1-2 in the preparation of nanomedicines for treating rheumatoid arthritis, characterized in that: The triptolide nanomedicine Y6@TPL is an injectable formulation, used in conjunction with photothermal therapy for synergistic treatment.

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