Bimetal nanoparticles as well as preparation method and application thereof
Through the self-assembly and disulfide bonding technology of bimetallic nanoparticles, copper ions are generated and GSH scavenging ability is enhanced, which solves the problem of difficult to improve tumor hypoxia microenvironment and induce copper death in the prior art, and achieves efficient tumor treatment and enhanced immune response.
Patent Information
- Application Number
- CN202411951402.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-27
- Publication Date
- 2025-05-13
AI Technical Summary
The prior art is difficult to effectively improve the hypoxic microenvironment of tumors and trigger copper death in tumor cells, limiting the efficacy of chemotherapy.
Bimetallic nanoparticles are used to form nanoclusters through self-assembly, and copper and gold or platinum are connected by disulfide bonds to generate copper ions and enhance glutathione (GSH) scavenging ability, thereby inducing tumor cell death and improving hypoxia by stably producing oxygen in response to the tumor microenvironment.
It has achieved effective induction of copper death in the tumor microenvironment, improved the efficiency of tumor treatment, improved the hypoxic microenvironment, and enhanced the anti-tumor immune response.
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Figure CN119970792A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of nanomaterials, and in particular relates to bimetallic nanoparticles and a preparation method and application thereof. Background Art
[0002] Bimetallic self-assembled nanomaterials, composed of atoms of two metals, decompose after the tumor microenvironment responds to improve the hypoxic microenvironment-induced copper death in the tumor site. They have attracted much attention from researchers due to their excellent tumor microenvironment-responsive decomposition ability and ion tumor delivery ability. At present, chemotherapy is still the main treatment for most cancers. Unfortunately, the complexity of the immunosuppressive tumor microenvironment and the low immunogenicity of cancer cells lead to reduced chemotherapy sensitivity, thus limiting their clinical efficacy. Recently, people have paid more and more attention to cancer immunotherapy aimed at activating and enhancing the innate immune system, and this field has received great attention in the scientific community. However, the immunosuppressive properties of tumors, marked by weakened immune responses, hinder the effective realization of the expected anti-tumor immunity. Therefore, people are increasingly interested in rebuilding this immunosuppressive tumor microenvironment. Recent studies have reported that inducing proteotoxic DLAT oligomerization can induce copper death of tumor cells through intracellular Cu accumulation. However, the hypoxic microenvironment of tumors is the main cause of tumor treatment resistance. A large number of studies have shown that improving hypoxia can effectively improve the treatment efficiency of tumors. At present, there is still no method that can effectively improve the hypoxic microenvironment of tumors and effectively induce copper death of tumor cells. Summary of the invention
[0003] In order to overcome at least one of the problems existing in the above-mentioned prior art, one of the objects of the present invention is to provide a bimetallic nanoparticle, which has the performance of improving hypoxia and inducing copper death at the same time, so that it has important application prospects in the preparation of tumor treatment materials.
[0004] A second object of the present invention is to provide a method for preparing the above-mentioned bimetallic nanoparticles.
[0005] The third object of the present invention is to provide an application of the above-mentioned bimetallic nanoparticles.
[0006] In order to achieve the above object, the technical solution adopted by the present invention is:
[0007] The first aspect of the present invention provides a bimetallic nanoparticle, which includes a nanocluster formed by self-assembly of nanoparticles; the nanoparticles include a first metal and a second metal; the first metal and the second metal are connected by a disulfide bond; the first metal includes copper; and the second metal includes gold or platinum.
[0008] In the present invention, the nanoclusters are composed of small nanoparticles, which are in a small particle state before self-assembly (just formed), and gradually form nanoclusters to form the final bimetallic nanoparticles.
[0009] Preferably, the particle size of the nanoparticles is in the range of 1 to 10 nm. In some specific embodiments of the present invention, the particle size of the nanoparticles is 3 nm, 5 nm, 7 nm or 10 nm.
[0010] Preferably, the particle size of the nanocluster is in the range of 100 to 200 nm, more preferably 110 to 150 nm, and even more preferably 110 to 140 nm. In some specific embodiments of the present invention, the particle size of the nanocluster is 110 nm, 120 nm, 130 nm or 140 nm.
[0011] Preferably, the surface of the nanoclusters is also modified with a polymer.
[0012] Preferably, the polymer comprises at least one of polyethylene glycol, polymaleic anhydride or octadecene-polymaleic anhydride-polyethylene glycol polymer (C18PMH-mPEG polymer).
[0013] The second aspect of the present invention provides a method for preparing bimetallic nanoparticles, comprising the following steps: mixing a first metal source, a second metal source, a disulfide, an amine compound, a dispersant and a solvent, performing a reduction reaction and self-assembly to obtain the bimetallic nanoparticles as described in the first aspect of the present invention; the first metal source includes a copper source; the first metal source includes a gold source or a platinum source.
[0014] Preferably, the disulfide comprises dithiodiglycolic acid, dithiodiglycolic acid salt or a combination thereof; further preferably, the disulfide is selected from dithiodiglycolic acid.
[0015] Preferably, the amine compound includes at least one of ethylamine, propylamine, ethylenediamine, triethylamine or hexamethylenediamine; further preferably, the amine compound is selected from triethylamine (TEA).
[0016] Preferably, the dispersant includes at least one of polyvinyl pyrrolidone, sodium dodecyl sulfonate, sodium dodecylbenzene sulfonate, polyvinyl alcohol or hexadecyltrimethylammonium bromide; further preferably, the dispersant is selected from polyvinyl pyrrolidone (PVP).
[0017] Preferably, the solvent includes at least one of ethanol, ethylene glycol, N,N-dimethylformamide or dimethyl sulfoxide; further preferably, the solvent includes ethanol and N,N-dimethylformamide (DMF).
[0018] Preferably, in the solvent, the volume ratio of ethanol to N,N-dimethylformamide is 1:(0.5-3); more preferably, it is 1:(1-2).
[0019] Preferably, the copper source comprises at least one of copper chloride, copper sulfate or copper nitrate; further preferably, the copper source is selected from copper chloride; further preferably, the copper source is selected from copper chloride dihydrate (CuCl 2 ·2H 2 O).
[0020] Preferably, the gold source comprises at least one of chloroauric acid, sodium chloroaurate or potassium chloroaurate; further preferably, the gold source is selected from chloroauric acid; further preferably, the gold source is selected from chloroauric acid trihydrate (HAuCl 4 ·3H 2 O).
[0021] Preferably, the platinum source comprises at least one of chloroplatinic acid, sodium chloroplatinate, potassium chloroplatinate or platinum nitrate; further preferably, the platinum source is selected from chloroplatinic acid; further preferably, the platinum source is selected from chloroplatinic acid hexahydrate (H 2 PtCl 6 6H 2 O).
[0022] Preferably, the molar ratio of the first metal source to the second metal source is 1:(0.5-2); more preferably, it is 1:(0.8-1.2).
[0023] Preferably, the molar ratio of the first metal source to the disulfide is 1:(0.5-2); more preferably, it is 1:(0.8-1.2).
[0024] Preferably, the molar ratio of the first metal source to the amine compound is 1:(0.2-3); more preferably, it is 1:(0.5-2).
[0025] Preferably, the usage ratio of the first metal source to the dispersant is 1 mol: (200-600) g; more preferably, it is 1 mol: (300-500) g.
[0026] Preferably, the usage ratio of the first metal source to the solvent is 1 mol:(10-30)L; more preferably, it is 1 mol:(15-25)L.
[0027] Preferably, after the reduction reaction and self-assembly, the product is centrifuged and washed.
[0028] Preferably, the washing reagent comprises at least one of ethanol, methanol or acetone.
[0029] Preferably, the reaction conditions of the reduction reaction and self-assembly are: reaction at 10-30° C. for 20-30 hours; or reaction at 120-160° C. for 4-8 hours.
[0030] The third aspect of the present invention provides a use of the bimetallic nanoparticles as described in the first aspect of the present invention, or the bimetallic nanoparticles prepared by the preparation method described in the second aspect of the present invention in the preparation of tumor treatment materials.
[0031] Preferably, the tumor therapeutic material exerts its effect by improving the hypoxic microenvironment of the tumor and / or inducing copper cell death in the tumor.
[0032] The beneficial effects of the present invention are as follows: in the bimetallic nanoparticles of the present invention, copper ions are generated when the disulfide bond is broken, and the copper ions can effectively induce tumor cell death, producing an excellent copper death effect; and, the first metal and the second metal are bridged by a disulfide bond, and the obtained nanoparticles have good glutathione (GSH) scavenging ability, and good GSH scavenging ability is conducive to enhancing the effect of copper and improving the effect of inducing tumor cell copper death; in addition, the bimetallic nanoparticles of the present invention can also effectively respond to the tumor microenvironment and stably and continuously generate oxygen, and have a good application effect in improving the tumor hypoxic microenvironment and inducing tumor cell copper death. BRIEF DESCRIPTION OF THE DRAWINGS
[0033] Figure 1 This is the TEM image of the ACS nanoparticles of Example 1.
[0034] Figure 2 This is a TEM image of the ACS nanoparticles in Example 1 during the self-assembly process.
[0035] Figure 3 This is the TEM image of the PCS nanoparticles of Example 3.
[0036] Figure 4 The ACS nanoparticles and H 2 O 2 Oxygen production curve.
[0037] Figure 5 The PCSP nanoparticles and H of Example 4 2 O 2 Oxygen production curve.
[0038] Figure 6 The results of protein imprinting of the ACSP nanoparticles of Example 2.
[0039] Figure 7 The figure shows the protein imprinting test results of the PCSP nanoparticles of Example 4. DETAILED DESCRIPTION
[0040] The content of the present invention is further described in detail below through specific examples. It should also be understood that the following examples are only used to further illustrate the present invention and cannot be construed as limiting the scope of protection of the present invention. Some non-essential improvements and adjustments made by those skilled in the art based on the principles set forth in the present invention all belong to the scope of protection of the present invention. The specific process parameters and the like in the following examples are also only examples within a suitable range, that is, those skilled in the art can make a selection within a suitable range through the description herein, and are not limited to the specific data exemplified below. The raw materials, reagents or devices used in the following examples and comparative examples, unless otherwise specified, can all be obtained from conventional commercial sources, or can be obtained by existing known methods.
[0041] In the following examples, room temperature refers to 10-30°C, more specifically 20-25°C.
[0042] Example 1
[0043] A bimetallic nanoparticle AuCuS (ACS), the specific preparation process is as follows:
[0044] First, HAuCl 4 ·3H 2 O (50 mM), CuCl 2 ·2H 2 O (50mM), polyvinyl pyrrolidone (PVP) 300mg, dithiodiglycolic acid (50mM) and triethylamine (TEA) 100-200μL were mixed in 15mL of a mixture of DMF and ethanol with a volume ratio of 5:3. Subsequently, self-assembly was performed by ultrasonic dispersion and magnetic stirring at room temperature for 24 hours. The final product was washed with ethanol and centrifuged to obtain self-assembled AuCuS (ACS) nanoparticles.
[0045] Example 2
[0046] A surface-modified bimetallic nanoparticle AuCuS-PEG (ACSP), the specific preparation process is as follows:
[0047] The ACS nanoparticles prepared in Example 1 were suspended in deionized water to obtain an ACS dispersion, 20 mg of octadecene-modified polymaleic anhydride-polyethylene glycol polymer (C18PMH-mPEG polymer) was added to 10 mL of the ACS dispersion, and the mixture was stirred at room temperature for 24 hours. Afterwards, the residual reactants were removed by centrifugation for purification to obtain surface-modified AuCuS-PEG (ACSP) nanoparticles.
[0048] Example 3
[0049] A bimetallic nanoparticle PtCuS (PCS), the specific preparation process is as follows:
[0050] First, H 2 PtCl 6 6H 2 O (50 mM), CuCl 2 ·2H 2 O (50mM), polyvinylpyrrolidone (PVP) 300mg, dithiodiglycolic acid (50mM) and triethylamine (TEA) 100-200μL were mixed in 15mL of a mixture of DMF and ethanol with a volume ratio of 5:3. Subsequently, after ultrasonic dispersion, it was transferred to a reactor and reacted at 140℃ for 6 hours for self-assembly. The final product was washed with ethanol and centrifuged to obtain self-assembled PtCuS (PCS) nanoparticles.
[0051] Example 4
[0052] A surface-modified bimetallic nanoparticle PtCuS-PEG (PCSP), the specific preparation process is as follows:
[0053] The PCS nanoparticles prepared in Example 3 were suspended in deionized water to obtain a PCS dispersion, 20 mg of octadecene-polymaleic anhydride-polyethylene glycol polymer (C18PMH-mPEG polymer) was added to 10 mL of the PCS dispersion, and the mixture was stirred at room temperature for 24 hours. Afterwards, the residual reactants were removed by centrifugation for purification to obtain surface-modified PtCuS-PEG (PCSP) nanoparticles.
[0054] Performance Testing
[0055] (1) The morphology of nanoparticles was observed using transmission electron microscopy (TEM).
[0056] Figure 1 This is the TEM image of the ACS nanoparticles of Example 1, which shows that the nanoparticles have uniform particle size and are uniform spherical particles. Figure 2 TEM images of the ACS nanoparticles of Example 1 during the self-assembly process, wherein (a) is the TEM image after 5 min of self-assembly, (b) is the TEM image after 30 min of self-assembly, and (c) is the TEM image after 60 min of self-assembly. The gradual aggregation of the nanoparticles during the self-assembly process can be observed, and it can be seen that the material synthesis process is a process of gradually self-assembling into large-sized nanoclusters.
[0057] Figure 3 This is the TEM image of the PCS nanoparticles of Example 3. It can be seen that the nanoparticles have uniform particle sizes and are uniform spherical particles, and the nanoparticles are composed of agglomerates of many small particles.
[0058] (2) Oxygen production performance measurement: At room temperature and pressure, 200 μg / mL of the nanoparticles of Examples 1 to 4 were mixed with 10 mM H2 O 2 During the co-incubation period, the oxygen production was measured using a dissolved oxygen meter; and the production of H alone was measured under the same conditions. 2 O 2 of oxygen production.
[0059] Figure 4 The ACS nanoparticles and H 2 O 2 The oxygen production curve shows that compared with H 2 O 2 The ACS nanoparticles can effectively catalyze H 2 O 2 Oxygen was produced, while no obvious oxygen was produced without the addition of ACS nanoparticles.
[0060] Figure 5 The PCSP nanoparticles and H of Example 4 2 O 2 The oxygen production curve shows that compared with H 2 O 2 The PCSP nanoparticles can effectively catalyze H 2 O 2 Oxygen is produced, while no obvious oxygen is produced without adding PCSP nanoparticles. And it can also be seen that good oxygen production effect can also be obtained after polymer modification.
[0061] (3) Test of the ability to induce copper death: A composite material of Elesclomol (a copper ion carrier) and Cu (ES+Cu group) was used as a positive control group, a control group without any treatment was used as a negative control group, and the nanoparticles of Examples 1 to 4 were used as experimental groups for protein imprinting test.
[0062] Figure 6 The protein imprint test results of ACSP nanoparticles in Example 2 are as follows: Figure 7 The protein imprint test results of the PCSP nanoparticles in Example 4 show that compared with the negative control group, the DLAT proteins of ACSP and PCSP nanoparticles are consistent with the positive control group, with an oligomerization band at 140KD, and the expression of FDX-1 protein is downregulated, that is, the FDX-1 proteins of the ACSP group, PCSP group and ES+Cu group are all downregulated, and the DLAT proteins are all oligomerized, indicating that ACSP and PCSP nanomaterials have the ability to induce copper death.
[0063] Through the preparation method in the embodiment of the present invention, a bimetallic self-assembled tumor microenvironment responsive nanoparticle can be obtained. The nanoparticle is first connected by gold or platinum and copper bimetallic through disulfide bonds to form nanoparticles of about 5nm, and then the bimetallic small nanoparticles self-assemble to form nanoclusters of about 120nm. The nanoclusters can effectively respond to the tumor microenvironment and stably and continuously produce oxygen, and can effectively induce copper death in tumor cells. Furthermore, coating a layer of polymer on the surface of the material can improve the biocompatibility of the material.
[0064] In summary, in the bimetallic nanoparticles of the present invention, copper ions are generated when the disulfide bond is broken, and the copper ions can effectively induce tumor cell death and produce excellent copper death effects; and the first metal and the second metal are bridged by a disulfide bond, and the obtained nanoparticles have good glutathione (GSH) scavenging ability, and good GSH scavenging ability is conducive to enhancing the role of copper and improving the effect of inducing tumor cell copper death; in addition, the bimetallic nanoparticles of the present invention can also effectively respond to the tumor microenvironment and stably and continuously produce oxygen, which has a good application effect in improving the tumor hypoxic microenvironment and inducing tumor cell copper death. In addition, the preparation method of the bimetallic nanoparticles of the present invention is simple, easy to operate, and the reaction conditions are mild, and it has broad application prospects in the fields of nanomedicine and tumor treatment.
Claims
1. A bimetallic nanoparticle, characterized in that: The bimetallic nanoparticles include nanoclusters formed by self-assembly of nanoparticles; the nanoparticles include a first metal and a second metal; the first metal and the second metal are connected by a disulfide bond; the first metal includes copper; and the second metal includes gold or platinum.
2. The bimetallic nanoparticles according to claim 1, characterized in that: The particle size of the nanoparticles ranges from 1 to 10 nm.
3. The bimetallic nanoparticles according to claim 1, characterized in that: The particle size of the nano clusters ranges from 100 to 200 nm.
4. The bimetallic nanoparticles according to claim 1, characterized in that: The surface of the nanoclusters is also modified with a polymer.
5. A method for preparing bimetallic nanoparticles, characterized in that: The method comprises the following steps: mixing a first metal source, a second metal source, a disulfide, an amine compound, a dispersant and a solvent, performing a reduction reaction and self-assembly, and obtaining the bimetallic nanoparticles as claimed in any one of claims 1 to 4; the first metal source comprises a copper source; the first metal source comprises a gold source or a platinum source.
6. The preparation method according to claim 5, characterized in that: The disulfide comprises dithiodiglycolic acid, dithiodiglycolic acid or a combination thereof; and / or, the amine compound comprises at least one of ethylamine, propylamine, ethylenediamine, triethylamine or hexamethylenediamine; and / or, the dispersant comprises at least one of polyvinyl pyrrolidone, sodium dodecyl sulfonate, sodium dodecylbenzene sulfonate, polyvinyl alcohol or hexadecyltrimethylammonium bromide; And / or, the solvent includes at least one of ethanol, ethylene glycol, N,N-dimethylformamide or dimethyl sulfoxide.
7. The preparation method according to claim 5, characterized in that: The copper source includes at least one of copper chloride, copper sulfate or copper nitrate; And / or, the gold source includes at least one of chloroauric acid, sodium chloroaurate or potassium chloroaurate; And / or, the platinum source includes at least one of chloroplatinic acid, sodium chloroplatinate, potassium chloroplatinate or platinum nitrate.
8. The preparation method according to claim 5, characterized in that: The molar ratio of the first metal source to the second metal source is 1:(0.5-2); and / or, the molar ratio of the first metal source to the disulfide is 1:(0.5-2); and / or, the molar ratio of the first metal source to the amine compound is 1:(0.2-3); and / or, the dosage ratio of the first metal source to the dispersant is 1 mol: (200-600) g; And / or, the usage ratio of the first metal source to the solvent is 1 mol:(10-30)L.
9. Use of the bimetallic nanoparticles according to any one of claims 1 to 4, or the bimetallic nanoparticles prepared by the preparation method according to any one of claims 5 to 8 in preparing tumor treatment materials.
10. The use according to claim 9, characterized in that: The tumor treatment material exerts its effects by improving the hypoxic microenvironment of the tumor and / or inducing copper death in the tumor.