Application of polydopamine-coated MOFs in preparation of medicine for inhibiting tumor metastasis
Polydopamine @MOFs were prepared by introducing defects into metal organic frame materials and loading polydopamine, which solved the problem of insufficient drug loading capacity of existing materials and achieved effective inhibition of tumor metastasis.
Patent Information
- Application Number
- CN202510165352.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-14
- Publication Date
- 2025-04-25
AI Technical Summary
The existing metal organic framework materials have insufficient drug-carrying capacity in inhibiting tumor metastasis, which is difficult to meet the needs of clinical applications.
By regulating defect sites with catechol during the synthesis of metal organic frame materials, a certain defective metal organic frame material is obtained, and polydopamine is loaded with this material to prepare polydopamine @MOFs.
Polydopamine@MOFs verified the synergistic physicochemical therapeutic effect at the cellular level, significantly inhibiting tumor metastasis, and showed effective inhibitory effects on breast cancer 4T1, melanoma B16F10 and non-small cell lung cancer lung-transforming agent models.
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Abstract
Description
Technical Field
[0001] The present invention relates to the field of medical technology, and in particular to an application of polydopamine@MOFs in the preparation of a drug for inhibiting tumor metastasis. Background Art
[0002] More than 90% of cancer patients die from tumor metastasis. Although chemotherapy, targeted therapy and immunotherapy are widely used in clinical practice, these treatments are not ideal in inhibiting tumor metastasis. Chemotherapeutic drugs are often toxic and easily lead to multidrug resistance (SR Krishnan, M. Bebawy, Mol Cancer. 2023, 22, 79-89). Although targeted therapy can reduce drug toxicity, it still faces many shortcomings, including drug resistance (AJSchoenfeld, MD Hellmann, Cancer Cell, 2020, 37, 443-55), high cost and insufficient drug exposure (M. Aldea, F. Andre, A. Marabelle, S. Dogan, F. Barlesi, J. Soria, Cancer Discov. 2021, 11, 874-899.) and other problems. However, immunotherapy faces problems such as cell tolerance and immunotoxicity (AV Finck, T. Blanchard, CP Roselle, G. Golinelli, CH June, Nat. Med. 2022, 28, 678-689). Developing new treatment methods and treatment systems to inhibit tumor metastasis is of great clinical significance.
[0003] Before metastasis, tumor cells will break through the basement membrane (R. Kalluri, Nat. Rev. Cancer, 2003, 3, 422-33.). Tumor cells will break through the basement membrane by physically squeezing the basement membrane or secreting matrix metalloproteinases MMP-2 / 9 to hydrolyze the basement membrane, or by using both methods (S. Linder, J. Cell Sci. 2009, 122, 3009-3013. J. Redondo-Mun, E. Ugarte-Berzal, MJ Terol, PESteen, MHCerro, M. Roderfeld, E. Roeb, G. Opdenakker, JA Garc1ˊa-Marco, AG Pardo, Cancer Cell, 2010, 17, 160-172) to achieve tumor metastasis.
[0004] Although existing studies have shown that metal-organic framework therapeutic systems can inhibit tumor metastasis, the drug-loading capacity of traditional metal-organic framework materials still needs to be further improved and is still difficult to meet the needs of clinical applications. Summary of the invention
[0005] In view of this, the purpose of the present invention is to provide an application of polydopamine@MOFs in the preparation of drugs for inhibiting tumor metastasis. The present invention uses catechol to regulate defect sites in the process of synthesizing metal organic framework materials, thereby obtaining metal organic framework materials with certain defects, and uses the prepared organic framework materials to load polydopamine to obtain polydopamine@MOFs. The polydopamine@MOFs of the present invention verifies the synergistic physical and chemical therapeutic effects in inhibiting tumor metastasis at the cellular level; in the breast cancer 4T1 lung metastasis model, the melanoma B16F10 lung metastasis model and the non-small cell lung cancer lung metastasis animal model, it is verified that the polydopamine@MOFs synergistic physical and chemical therapeutic effects inhibit tumor metastasis.
[0006] The present invention is specifically implemented by adopting the following technical solutions:
[0007] A use of polydopamine@MOFs in preparing a drug for inhibiting tumor metastasis, wherein the tumor metastasis includes breast cancer 4T1 lung metastasis and melanoma B16F10 lung metastasis.
[0008] The preparation method of polydopamine@MOFs of the present invention comprises the following steps:
[0009] (1) Synthesis of metal-organic framework materials rich in defect sites
[0010] Add 100 mg of 2,3,6,7,10,11-hexahydroxytriphenyl (CAS No. 4877-80-9), 92 mg of ferrous sulfate heptahydrate, 100 mg of tetrabutylammonium nitrate, 5-15 mg of catechol and 5.6 mL of solvent into a reactor, add 45 μL of pentylamine and stir evenly, then react the reactor at 150-200° C. for 30-50 hours, cool and centrifuge to obtain metal organic framework materials MOFs with defects of 4.3%-8.2%;
[0011] (2) Activation of metal-organic framework materials rich in defect sites
[0012] The metal organic framework material prepared in step (1) is immersed in anhydrous DMF for 2 days, and the solution is replaced with fresh solution every 12 hours; then immersed in anhydrous acetone for 4 days, and the solution is replaced with fresh solution every 12 hours, and finally dried in vacuum at room temperature;
[0013] (3) Preparation of polydopamine@MOFs
[0014] 1 g of activated MOFs was dispersed in anhydrous ethanol, and then 1 g of polydopamine was added. The mixture was stirred at room temperature overnight, and then the solid was collected by centrifugation. After washing with ethanol, the solid was vacuum dried to obtain polydopamine@MOFs.
[0015] Preferably, the solvent in step (1) is a mixture of N,N-diethylformamide, methanol and water in a volume ratio of 5:0.3:0.3.
[0016] Preferably, in step (1), the reaction temperature is 180° C. and the reaction time is 42 h.
[0017] Preferably, in step (3), the centrifugal speed is 11000 rpm and the centrifugal time is 30 min.
[0018] Preferably, the vacuum drying temperature in step (3) is 50° C. and the drying time is 48 h.
[0019] Compared with the prior art, the present invention has the following beneficial effects:
[0020] The polydopamine@MOFs of the present invention has a synergistic physical and chemical therapeutic effect to inhibit tumor metastasis, including: 1) using small molecules that inhibit the activity of metalloproteinases as organic ligands to construct MOFs with metal clusters, and regulating their defects; 2) MOFs are equipped with polydopamine monomers to construct a synergistic physical and chemical therapeutic system; 3) polydopamine@MOFs verifies the synergistic physical and chemical therapeutic effect to inhibit tumor metastasis at the cellular level; 4) in breast cancer 4T1 lung metastasis model, melanoma B16F10 lung metastasis model and non-small cell lung cancer lung metastasis animal model, the synergistic physical and chemical therapeutic effect of polydopamine@MOFs is verified to inhibit tumor metastasis. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 This is the synthesis step of the polydopamine@MOFs with synergistic physicochemical therapeutic effects of the present invention.
[0022] Figure 2 are the X-ray powder diffraction patterns of MOFs-8.2% and polydopamine@MOFs-8.2%.
[0023] Figure 3 These are the N2 adsorption graphs of MOFs-8.2% and polydopamine@MOFs-8.2%.
[0024] Figure 4 are the particle size distribution and Zeta potential of MOFs-8.2% and polydopamine@MOFs-8.2%.
[0025] Figure 5 Transmission electron microscopy images of MOFs-8.2% (B) and polydopamine@MOFs-8.2% (A).
[0026] Figure 6 Figure 3. The release of polydopamine monomers from MOFs-8.2% and polydopamine@MOFs with different defects in response to the acidic microenvironment of tumors.
[0027] Figure 7 This is a representative schematic diagram (A) and corresponding statistical graph (B) showing that polydopamine@MOFs-8.2% of the present invention inhibits the migration of B16F10 tumor cells in an in vitro experiment.
[0028] Figure 8 This is a representative schematic diagram (A) and corresponding statistical diagram (B) of the in vitro experiment showing that the polydopamine@MOFs-8.2% synergistic physical and chemical therapeutic effects of the present invention inhibit the migration of 4T1 tumor cells.
[0029] Fig. 9 In an in vivo mouse experiment, the polydopamine@MOFs-8.2% of the present invention synergized the physical and chemical therapeutic effects to inhibit the migration of 4T1 tumor cells. Schematic diagram of the experimental procedure (A); representative H&E staining schematic diagram (B) and corresponding statistical diagram (C).
[0030] Fig.10 In an in vivo mouse experiment, the polydopamine@MOFs-8.2% of the present invention synergized the physical and chemical therapeutic effects to inhibit the migration of non-small cell lung cancer (LLC) cells and treat tumor metastasis. Schematic diagram of the experimental procedure (A); representative H&E staining schematic diagram (B) and corresponding statistical graph (C). DETAILED DESCRIPTION
[0031] The synthesis method of polydopamine@MOFs of the present invention is as follows Figure 1 As shown, the following steps are included:
[0032] (1) Synthesis of metal-organic framework materials rich in defect sites
[0033] 100 mg 2,3,6,7,10,11-hexahydroxytriphenyl (abbreviated as HTP), 92 mg ferrous sulfate heptahydrate, 100 mg tetrabutylammonium nitrate, 5-15 mg catechol and 5.6 mL solvent were added to the reactor, 45 μL pentylamine was added and stirred evenly, and then the reactor was reacted at 150-200° C. for 30-50 hours, cooled and centrifuged to obtain metal organic framework materials MOFs with defects of 4.3% to 8.2%; in some embodiments, MOFs with defects of 4.3%, 6.6% and 8.2% were obtained, which were respectively expressed as MOF-4.3%, -6.6% and -8.2%; the characterization method of the defect rate of MOFs materials of the present invention is obtained by elemental analysis, specifically, elemental analysis is performed on the synthesized materials to obtain the type and content information of the elements. Compared with the theoretical chemical formula of the material, the content of the missing element is calculated to quantify the percentage of defects.
[0034] (2) Activation of metal-organic framework materials rich in defect sites
[0035] The metal organic framework material prepared in step (1) is immersed in anhydrous DMF for 2 days, and the solution is replaced with fresh solution every 12 hours; then immersed in anhydrous acetone for 4 days, and the solution is replaced with fresh solution every 12 hours, and finally dried in vacuum at room temperature;
[0036] (3) Preparation of polydopamine@MOFs
[0037] 1 g of activated MOFs was dispersed in anhydrous ethanol, and then 1 g of polydopamine was added. The mixture was stirred at room temperature overnight, and then the solid was collected by centrifugation. After washing with ethanol, the solid was vacuum dried to obtain polydopamine@MOFs.
[0038] The solvent in step (1) of the present invention is a mixture of N,N-diethylformamide, methanol and water in a volume ratio of 5:0.3:0.3.
[0039] In step (1) of the present invention, the reaction temperature is 180° C. and the reaction time is 42 h.
[0040] In step (3) of the present invention, the centrifugal speed is 11000 rpm and the centrifugal time is 30 min.
[0041] The vacuum drying temperature in step (3) of the present invention is 50° C. and the drying time is 48 hours.
[0042] In order to make the purpose, technical scheme and advantages of the present invention clearer, the preferred embodiments of the present invention are further described in detail in conjunction with the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0043] Example 1
[0044] A synthesis method of polydopamine@MOFs is as follows:
[0045] (1) Synthesis of metal-organic framework materials rich in defect sites
[0046] 100 mg of 2,3,6,7,10,11-hexahydroxytriphenyl, 92 mg of ferrous sulfate heptahydrate, 100 mg of tetrabutylammonium nitrate, 5 mg of catechol and 5.6 mL of solvent (mixed by 5 mL of N,N-diethylformamide, 0.3 mL of methanol and 0.3 mL of water) were added to the reactor, 45 μL of pentylamine was added under stirring and stirred evenly, and then the reactor was reacted at 180° C. for 42 h, cooled and centrifuged to obtain a metal organic framework material MOFs with a defect of 4.3%, namely MOF-4.3%;
[0047] (2) Activation of metal-organic framework materials rich in defect sites
[0048] The MOF-4.3% prepared in step (1) was immersed in anhydrous DMF for 2 days, and the solution was replaced with fresh solution every 12 hours; then immersed in anhydrous acetone for 4 days, and the solution was replaced with fresh solution every 12 hours, and finally dried under vacuum at room temperature;
[0049] (3) Preparation of polydopamine@MOFs-4.3%
[0050] 1 g of activated MOFs was dispersed in anhydrous ethanol, and then 1 g of polydopamine was added. The mixture was stirred at room temperature overnight, and then centrifuged (centrifugal speed was 11000 rpm, and centrifugal time was 30 min) to collect the solid. After washing with ethanol, the solid was vacuum dried at 50°C for 48 h to obtain polydopamine@MOFs-4.3%.
[0051] Example 2
[0052] A synthesis method of polydopamine@MOFs is as follows:
[0053] (1) Synthesis of metal-organic framework materials rich in defect sites
[0054] 100 mg of 2,3,6,7,10,11-hexahydroxytriphenyl, 92 mg of ferrous sulfate heptahydrate, 100 mg of tetrabutylammonium nitrate, 10 mg of catechol and 5.6 mL of solvent (mixed by 5 mL of N,N-diethylformamide, 0.3 mL of methanol and 0.3 mL of water) were added to the reactor, 45 μL of pentylamine was added under stirring and stirred evenly, and then the reactor was reacted at 180° C. for 42 h, cooled and centrifuged to obtain a metal organic framework material MOFs with a defect rate of 6.6%, namely MOF-6.6%;
[0055] (2) Activation of metal-organic framework materials rich in defect sites
[0056] The MOF-6.6% prepared in step (1) was immersed in anhydrous DMF for 2 days, and the solution was replaced with fresh solution every 12 hours; then immersed in anhydrous acetone for 4 days, and the solution was replaced with fresh solution every 12 hours, and finally dried under vacuum at room temperature;
[0057] (3) Preparation of polydopamine@MOFs-6.6%
[0058] 1 g of activated MOFs was dispersed in anhydrous ethanol, and then 1 g of polydopamine was added. The mixture was stirred at room temperature overnight, and then centrifuged (centrifugal speed was 11000 rpm, and centrifugal time was 30 min) to collect the solid. After washing with ethanol, the solid was vacuum dried at 50°C for 48 h to obtain polydopamine@MOFs-6.6%.
[0059] Example 3
[0060] A synthesis method of polydopamine@MOFs is as follows:
[0061] (1) Synthesis of metal-organic framework materials rich in defect sites
[0062] 100 mg of 2,3,6,7,10,11-hexahydroxytriphenyl, 92 mg of ferrous sulfate heptahydrate, 100 mg of tetrabutylammonium nitrate, 15 mg of catechol and 5.6 mL of solvent (mixed by 5 mL of N,N-diethylformamide, 0.3 mL of methanol and 0.3 mL of water) were added to the reactor, 45 μL of pentylamine was added under stirring and stirred evenly, and then the reactor was reacted at 180° C. for 42 h, cooled and centrifuged to obtain a metal organic framework material MOFs with a defect of 8.2%, namely MOF-8.2%;
[0063] (2) Activation of metal-organic framework materials rich in defect sites
[0064] The MOF-8.2% prepared in step (1) was immersed in anhydrous DMF for 2 days, and the solution was replaced with fresh solution every 12 hours; then immersed in anhydrous acetone for 4 days, and the solution was replaced with fresh solution every 12 hours, and finally dried under vacuum at room temperature;
[0065] (3) Preparation of polydopamine@MOFs-8.2%
[0066] 1 g of activated MOFs was dispersed in anhydrous ethanol, and then 1 g of polydopamine was added. The mixture was stirred at room temperature overnight, and then centrifuged (centrifugal speed was 11000 rpm, and centrifugal time was 30 min) to collect the solid. After washing with ethanol, the solid was vacuum dried at 50°C for 48 h to obtain polydopamine@MOFs-8.2%.
[0067] Figure 2 to Figure 5 They are the X-ray powder diffraction pattern, N2 adsorption pattern, particle size distribution, Zeta potential and transmission electron microscopy pattern of polydopamine@MOFs-8.2% and MOF-8.2% prepared in Example 3 respectively.
[0068] 1. The dopamine release rate of polydopamine@MOFs-4.3%, polydopamine@MOFs-6.6%, and polydopamine@MOFs-8.2% prepared in Examples 1 to 3 was tested as follows:
[0069] Polydopamine@MOFs with different defects were dispersed in a solution with pH = 6.5, and the supernatant was taken at intervals. The content was tested using a UV-Vis spectrophotometer to draw a time-release curve. Figure 6It can be seen that under the condition of pH=6.5 in the tumor microenvironment, the greater the defect content, the faster the release rate, the more dopamine released, and the best therapeutic effect. The polydopamine@MOFs-8.2% of the present invention has the fastest release rate.
[0070] 2. Cell experiments verified the inhibitory effect of polydopamine@MOFs-8.2% synergistic physicochemical effect on tumor cell invasion.
[0071] Transwell experiments were used to verify that polydopamine@MOFs-8.2% synergized the regulation of dopamine multi-basement membrane viscosity and the ligand HTP's inhibition of matrix metalloproteinases to overcome tumor metastasis, as follows:
[0072] Using an 8-micron Boyden chamber, 70 microliters of matrigel (simulating basement membrane) was first spread in the upper chamber, and PBS, HTP small molecules, metal cluster salts, single polydopamine, MOF-8.2% without polydopamine, and polydopamine@MOFs-8.2% were added respectively. After pre-fixation in an incubator for half an hour, 10,000 200-microliter serum-free 4T1 breast or B16F10 melanoma tumor cells were planted on it (600 microliters of culture medium containing 10% fetal bovine serum was added to the lower chamber. After culturing in the incubator for 24 hours, 4% paraformaldehyde was added to the upper chamber for fixation and stained with 0.1% crystal violet. The invading cells at the bottom of the upper chamber can be observed and counted under a microscope, such as Figure 7 (Breast tumor) or Figure 8 (Melanoma), it can be seen that compared with other control groups, polydopamine@MOFs-8.2% can significantly inhibit the invasion of breast 4T1 and melanoma B16F10 tumor cells, proving the effectiveness of polydopamine@MOFs-8.2% in treating tumors.
[0073] 3. Animal experiments verified the inhibitory effect of polydopamine@MOFs-8.2% synergistic physicochemical effect on tumor cell invasion.
[0074] In order to prove in vivo that polydopamine@MOFs-8.2% can synergistically inhibit tumor cell invasion through physical and chemical effects, two spontaneous tumor metastasis models were constructed, including breast cancer lung metastasis and non-small cell lung metastasis models. The steps for constructing the lung metastasis model are as follows: for the breast cancer model, 500,000 4T1 tumor cells were implanted in the fourth mammary pad of female BALB / C mice aged 6 to 8 weeks; for the non-small cell lung cancer model, 300,000 B16F10 cells were implanted subcutaneously on the back of C57BL / 6J mice aged 6 to 8 weeks. When the tumor volume reached 60 cubic millimeters, the mice were randomly divided into 6 groups, with 5 mice in each group. Then, every 3 days, the treatment group was injected with the designated reagents intratumorally. At the end of the treatment, the mice were killed, and the lungs were removed for paraffin sectioning and H&E staining, and the metastatic lesions could be quantitatively counted and analyzed. The results show that Fig. 9 and Fig.10 : HTP or polydopamine alone has only limited therapeutic effect, while polydopamine@MOFs-8.2% can significantly inhibit tumor metastasis, with an inhibition efficiency of 86.5% for breast cancer metastasis and 97.2% for non-small cell lung cancer metastasis, demonstrating the synergistic physicochemical therapeutic effect of polydopamine@MOFs-8.2% in tumor metastasis.
[0075] The embodiments described above are part of the embodiments of the present invention, rather than all of the embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
Claims
1. Application of polydopamine@MOFs in the preparation of drugs for inhibiting tumor metastasis.
2. The use according to claim 1, characterized in that: The preparation method of polydopamine@MOFs comprises the following steps: (1) Synthesis of metal-organic framework materials rich in defect sites Add 100 mg of 2,3,6,7,10,11-hexahydroxytriphenyl, 92 mg of ferrous sulfate heptahydrate, 100 mg of tetrabutylammonium nitrate, 5-15 mg of catechol and 5.6 mL of solvent into a reactor, add 45 μL of pentylamine and stir evenly, then react the reactor at 150-200° C. for 30-50 h, cool and centrifuge to obtain metal organic framework materials MOFs with defects of 4.3%-8.2%; (2) Activation of metal-organic framework materials rich in defect sites The metal organic framework material prepared in step (1) is immersed in anhydrous DMF for 2 days, and the solution is replaced with fresh solution every 12 hours; then immersed in anhydrous acetone for 4 days, and the solution is replaced with fresh solution every 12 hours, and finally dried in vacuum at room temperature; (3) Preparation of polydopamine@MOFs 1 g of activated MOFs was dispersed in anhydrous ethanol, and then 1 g of polydopamine was added. The mixture was stirred at room temperature overnight, and then the solid was collected by centrifugation. After washing with ethanol, the solid was vacuum dried to obtain polydopamine@MOFs.
3. The use according to claim 2, characterized in that: The solvent in step (1) is a mixture of N,N-diethylformamide, methanol and water in a volume ratio of 5:0.3:0.
3.
4. The use according to claim 2, characterized in that: In step (1), the reaction temperature is 180° C. and the reaction time is 42 h.
5. The use according to claim 2, characterized in that: In step (3), the centrifugal speed is 11000 rpm and the centrifugal time is 30 min.
6. The use according to claim 2, characterized in that: The vacuum drying temperature in step (3) is 50° C. and the drying time is 48 h.
7. The use according to claim 1, characterized in that: The tumor metastases include breast cancer 4T1 lung metastasis and melanoma B16F10 lung metastasis.