Palladium-copper-cobalt nano material and preparation method and application thereof

Abstract

The invention relates to a palladium-copper-cobalt nanometer material and preparation and application thereof, the nanometer material comprises palladium metal, copper metal and cobalt metal, the molar ratio of the palladium metal to the copper metal to the cobalt metal is 1: (7-8): (15-17), and the nanometer material is of a layered structure. The invention has the advantages of favorable biocompatibility and lower biotoxicity, and also has favorable inhibition effect and treatment effect on breast cancer.

Description

Technical Field

[0001] This invention relates to the field of tumor drug technology, specifically to a palladium-copper-cobalt nanomaterial, its preparation method, and its application. Background Technology

[0002] Currently, commonly used anti-tumor drugs, especially those for treating breast cancer, mainly include immune checkpoint inhibitors (such as PD-1 / PD-L1 antibodies), antibody-drug conjugates (ADCs), and small molecule targeted drugs. Although these drugs have shown efficacy in various tumors, they still face the following common challenges: (1) Drug resistance: Tumor cells can develop drug resistance through mechanisms such as gene mutation and phenotypic transformation, leading to treatment failure; (2) Toxicity and side effects: Existing therapies often use the maximum tolerated dose (MTD) for administration, resulting in immune-related adverse reactions, hematological toxicity, etc., affecting patients' quality of life and treatment compliance; (3) Tumor microenvironment (TME) disorders: Solid tumors generally have hypoxia, high glutathione (GSH) levels, and an acidic microenvironment. These characteristics not only inhibit the function of immune cells but also weaken the efficacy of chemotherapy, radiotherapy, and photodynamic therapy by scavenging reactive oxygen species (ROS). Existing drugs are difficult to effectively regulate the TME, constituting a "ceiling" for efficacy. Summary of the Invention

[0003] The purpose of this invention is to overcome the shortcomings of the prior art and provide a palladium-copper-cobalt nanomaterial.

[0004] The purpose of this application is also to provide a method for preparing and applying the above-mentioned palladium-copper-cobalt nanomaterials.

[0005] To achieve the objectives of this invention, the following technical solutions are provided in this application.

[0006] In a first aspect, this application provides a palladium-copper-cobalt nanomaterial comprising palladium, copper, and cobalt metals in a molar ratio of 1:(7-8):(15-17), and the nanomaterial having a layered structure. At the electronic structure level, Co acts as an electron donor, undergoing directional charge transfer to Pd and Cu (Bader charge analysis shows that Pd gains electrons, and Co / Cu loses electrons), causing the d-band center of Pd to shift upwards to the optimal adsorption energy region. This enhances the adsorption and activation capacity for substrates such as H2O2 and O2, while avoiding catalyst poisoning caused by excessive adsorption of intermediate products; Cu utilizes its Cu... + / Cu 2+The variable valence state constructs a sustainable Fenton reaction cycle, providing a redox switch for the continuous generation of reactive oxygen species (ROS). Specifically, through a unique multi-metal synergistic effect, it simultaneously mimics the activities of catalase (CAT), glutathione peroxidase (GPx), peroxidase (POD), and oxidase (OXD), achieving multiple regulation of tumor hypoxia relief, GSH depletion, and efficient ROS generation.

[0007] In one embodiment of the first aspect, the particle size of the nanomaterial is 300~320 nm.

[0008] In one embodiment of the first aspect, the thickness of each layer of the nanomaterial is 0.6-0.9 nm. This thickness maximizes catalytic activity. The atomic-level thickness allows for a surface atomic ratio exceeding 80%, exposing almost all Pd, Cu, and Co atoms as catalytically active sites. Simultaneously, the ultrathin structure leads to coordination unsaturation, forming defects such as lattice distortion and amorphous regions. These defect sites possess higher surface energies, preferentially adsorbing substrate molecules such as H₂O₂ and O₂, significantly reducing the reaction activation energy.

[0009] In one embodiment of the first aspect, the nanomaterial is modified with PVP by electrostatic adsorption, wherein the mass ratio of the nanomaterial to PVP is 1:15~30. The PVP coating forms a spatial barrier on the surface, preventing the nanosheets from aggregating in the physiological environment and improving dispersion stability; PVP reduces protein adsorption, lowers immune recognition, and prolongs in vivo circulation time, while not blocking active sites, thus maintaining catalytic activity.

[0010] In a second aspect, this application also provides a method for preparing the above-mentioned palladium-copper-cobalt nanomaterials. The method includes the following steps: dissolving a palladium source, a copper source, a cobalt source, tungsten hexacarbonyl, and a reducing agent in a solvent, and sequentially subjecting the mixture to ultrasonic treatment, heating, cooling, centrifugation, washing, and drying to obtain the nanomaterials. Utilizing the mild reducing properties of ascorbic acid, in a weakly acidic environment regulated by acetic acid, and using the CO gas released by the thermal decomposition of W(CO)6 as a morphology control agent, the three acetylacetone metal salts are co-reduced, thereby inducing ultrathin PdCuCo ternary alloy nanosheets with lattice distortion.

[0011] In one embodiment of the second aspect, the palladium source is palladium diacetylacetonate, the copper source is copper diacetylacetonate, the cobalt source is cobalt diacetylacetonate, the reducing agent is ascorbic acid, and the molar ratio of the palladium source, copper source, cobalt source, tungsten hexacarbonyl and reducing agent is (1.0~1.1):(1.2~1.3):1.0:(5.4~5.6):(7.2~7.4). The solvent is a mixed solution of N,N-dimethylformamide and acetic acid, and the volume ratio of N,N-dimethylformamide to acetic acid is (3~5):1.

[0012] In one embodiment of the second aspect, the preparation method includes at least one of the following features: 1) The ultrasonic treatment is performed at room temperature for 0.5–2 hours. Room temperature ensures a mild and moderate cavitation effect, sufficient to break down precursor aggregates while preventing premature decomposition of heat-sensitive precursors (especially W(CO)6). W(CO)6 begins to decompose at temperatures above 60°C. If the ultrasonic process involves excessively high temperatures, CO will be released prematurely, resulting in a lack of CO templates during the subsequent oil bath treatment, thus preventing the formation of two-dimensional sheet-like structures.

[0013] 2) The heating is carried out in an oil bath at a temperature of 60-100℃ for 8-15 hours. The decomposition temperature of W(CO)6 is approximately 60℃. Below this temperature, very little CO is released, insufficient bubble templates are formed, and lattice distortion is difficult to introduce, resulting in products that tend to be thick particles rather than ultrathin nanosheets. Simultaneously, the reduction rate of ascorbic acid at this temperature is too slow, requiring a significantly extended reaction time (>24 hours), leading to low efficiency and easy introduction of impurities. At this temperature, the moderate decomposition of W(CO)6 continuously provides CO templates, the ascorbic acid reduction rate is moderate, and the crystals grow slowly along the two-dimensional direction, ultimately forming nanosheets with atomic-level thickness.

[0014] 3) The cooling is natural cooling, cooling to room temperature; 4) The centrifugation speed is 6000~10000 rpm; the centrifugation time is 5~20 min; 5) The washing process uses ethanol and involves at least 5 washes; 6) The drying temperature is 40~60℃ and the drying time is 6~8h.

[0015] In one embodiment of the second aspect, the product is dissolved, modified, washed a second time, and dried to obtain the nanomaterial. This operation is mainly to obtain powdered particles for easy weighing and quantification.

[0016] In one embodiment of the second aspect, the preparation method includes at least one of the following features: a) The dissolution is achieved by deionizing the product solution in water; b) The modification involves adding PVP and then performing magnetic stirring, wherein the mass ratio of the nanomaterial to PVP is 1:(15~30), the stirring rate of the magnetic stirring is 100~200 rpm, and the stirring time is 20~30 h; c) The secondary washing is performed by alternating washing with deionized water and anhydrous ethanol, and the number of washing cycles is not less than 3. d) The drying temperature is 40~60℃ and the drying time is 6~8h.

[0017] In a third aspect, this application also provides an application of the palladium-copper-cobalt nanomaterial as described above, said nanomaterial being used to prepare an injection for treating breast cancer.

[0018] In one embodiment of the third aspect, the active ingredient of the injection is the nanomaterial, the solvent of the injection is phosphate buffer, and the concentration of the nanomaterial is 15-25 mg / kg. More preferably, the concentration of the nanomaterial is 20 mg / kg.

[0019] In one embodiment of the third aspect, the tumor site is subjected to ultrasound manipulation after the injection.

[0020] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) The material of this application achieves a surface atomic ratio of >80% through a two-dimensional structure design with an atomic thickness (<1 nm), maximizing the exposure of active sites; at the same time, by utilizing the electronic synergistic effect of Pd, Cu and Co metals, a triple killing mechanism of "alleviating hypoxia + depleting GSH + efficient ROS production" is constructed, which can effectively inhibit the development of breast cancer. (2) The introduction of ultrasound as an external physical field trigger enables spatiotemporal controllable enhancement of catalytic activity and significantly reduces systemic toxicity. In vivo safety evaluation confirmed that no significant abnormalities were found in liver and kidney function and histology of major organs in mice after intravenous injection of PCC, indicating excellent biocompatibility. Experimental data showed that in the 4T1 breast cancer model, the tumor inhibition rate of the PCC+US group reached 64%, and the number of lung metastatic nodules was reduced by 9.8 times compared with the control group. Attached Figure Description

[0021] Figure 1 The image shows the full X-ray photoelectron spectrum of the final product of Example 1.

[0022] Figure 2 This is an atomic force microscope image of the final product of Example 1.

[0023] Figure 3 The height distribution curve is the atomic force microscope image corresponding to the final product of Example 1.

[0024] Figure 4 This is an elemental distribution diagram of Pd, Cu, and Co in the final product of Example 1, obtained by energy dispersive spectroscopy under a transmission electron microscope.

[0025] Figure 5 The graph shows the changes in body weight of four groups of mice within 14 days after injection of PCC solutions of different concentrations.

[0026] Figure 6 The images show the hematoxylin-eosin (H&E) staining results of four groups of mice after injection of PCC solutions of different concentrations.

[0027] Figure 7 The results of changes in body weight of mice in each group are shown in the examples of tumor treatment efficacy.

[0028] Figure 8 The results of tumor volume changes in mice in each group are shown in the examples of tumor treatment efficacy.

[0029] Figure 9 The results of changes in lung tissue weight in mice in each group are shown in the tumor treatment efficacy examples. Detailed Implementation

[0030] Unless otherwise stated, implied from the context, or as is customary in the art, all parts and percentages in this application are based on weight, and all testing and characterization methods used are concurrent with the filing date of this application. Where applicable, any patent, patent application, or disclosure relating to this application is incorporated herein by reference in its entirety, and its equivalent patent families are also incorporated herein by reference, particularly the definitions disclosed in these documents concerning synthetic techniques, product and processing design, polymers, comonomers, initiators, or catalysts in the art. If any definition of a specific term disclosed in the prior art is inconsistent with any definition provided in this application, the definition provided in this application shall prevail.

[0031] The numerical ranges in this application are approximate values ​​and therefore may include values ​​outside the range unless otherwise stated. A numerical range includes all values ​​from the lower limit to the upper limit, increasing by one unit, provided there is an interval of at least two units between any lower and any higher value. For example, if a component, physical, or other property (such as molecular weight) is described as 100 to 1000, this means that all individual values, such as 100, 101, 102, etc., are explicitly listed, as well as all subranges, such as 100 to 166, 155 to 170, 198 to 200, etc. For ranges containing values ​​less than 1 or fractions greater than 1 (e.g., 1.1, 1.5, etc.), one unit is appropriately considered as 0.0001, 0.001, 0.01, or 0.1. For ranges containing single digits less than 10 (e.g., 1 to 5), one unit is generally considered as 0.1. These are merely specific examples of what is intended to be expressed, and all possible combinations of values ​​between the listed minimum and maximum values ​​are considered to be clearly stated in this application. It should also be noted that the terms "first," "second," etc., used herein are not intended to specify a particular order, but are merely used to distinguish substances with different structures.

[0032] When referring to chemical compounds, unless explicitly stated otherwise, the singular includes all isomers and vice versa (e.g., "hexane" includes all isomers of hexane, individually or collectively). Additionally, unless explicitly stated otherwise, nouns described with "an," "a," or "the" also include their plural forms.

[0033] The terms “comprising,” “including,” “having,” and their derivatives do not exclude the presence of any other components, steps, or processes, regardless of whether such other components, steps, or processes are disclosed in this application. To eliminate any doubt, unless expressly stated otherwise, all compositions using the terms “comprising,” “including,” or “having” in this application may contain any additional additives, excipients, or compounds. Conversely, except for those necessary for operational performance, the term “substantially constitutes…” excludes any other components, steps, or processes described below with respect to that term. The term “consisting of…” does not include any components, steps, or processes not specifically described or listed. Unless expressly stated otherwise, the term “or” refers to the individual members listed or any combination thereof. Example

[0034] The embodiments of the present invention will be described in detail below. These embodiments are implemented based on the technical solution of the present invention, and detailed implementation methods and specific operation processes are given. However, the scope of protection of the present invention is not limited to the following embodiments. Example

[0035] Palladium-copper-cobalt (PCC) nanomaterials were synthesized using a wet chemical method, with the following steps: 10 mg of palladium diacetylacetonate, 10 mg of copper diacetylacetonate, 8 mg of cobalt diacetylacetonate, 60 mg of tungsten hexacarbonyl, and 40 mg of ascorbic acid were dissolved in a mixed solvent of 16 mL of N,N-dimethylformamide and 4 mL of acetic acid. The solution was sonicated at below 25 °C for 1 hour, then transferred to a sealed glass bottle and heated in an oil bath at 80 °C for 12 hours. After the system cooled naturally to room temperature, the product was centrifuged (8000 rpm), washed six times with ethanol, and finally dried at 60 °C to obtain the crude product. 10 mg of the crude product was dissolved in deionized water, and then 200 mg of PVP was added to the solution. The mixture was magnetically stirred for 24 hours. Subsequently, the obtained product was washed three times alternately with deionized water and anhydrous ethanol, and dried to obtain the final product (PCC). Example

[0036] Palladium-copper-cobalt nanomaterials (PCC) were synthesized using a wet chemical method, with the following specific steps: 10 mg of palladium diacetylacetonate, 10 mg of copper diacetylacetonate, 8 mg of cobalt diacetylacetonate, 60 mg of tungsten hexacarbonyl, and 40 mg of ascorbic acid were dissolved in a mixed solvent of 15 mL N,N-dimethylformamide and 5 mL acetic acid. The solution was sonicated at below 25 °C for 0.5 h, then transferred to a sealed glass bottle and heated in an oil bath at 60 °C for 15 h. After the system cooled naturally to room temperature, the product was centrifuged (6000 rpm), washed five times with ethanol, and finally dried at 40 °C for 4 h to obtain the crude product. 10 mg of the crude product was dissolved in deionized water, and then 150 mg of PVP was added to the solution. The mixture was magnetically stirred for 30 h. Subsequently, the obtained product was washed three times alternately with deionized water and anhydrous ethanol, and dried at 40 °C to obtain the final product (PCC). Example

[0037] Palladium-copper-cobalt nanomaterials (PCC) were synthesized using a wet chemical method, with the following specific steps: 10 mg of palladium diacetylacetonate, 10 mg of copper diacetylacetonate, 8 mg of cobalt diacetylacetonate, 60 mg of tungsten hexacarbonyl, and 40 mg of ascorbic acid were dissolved in a mixed solvent of 20 mL N,N-dimethylformamide and 4 mL acetic acid. The solution was sonicated at below 25 °C for 2 hours, then transferred to a sealed glass bottle and heated in an oil bath at 100 °C for 8 hours. After the system cooled naturally to room temperature, the product was centrifuged (10,000 rpm), washed six times with ethanol, and finally dried at 50 °C for 7 hours to obtain the crude product. 10 mg of the crude product was dissolved in deionized water, and then 300 mg of PVP was added to the solution. The mixture was magnetically stirred for 20 hours. Subsequently, the obtained product was washed three times alternately with deionized water and anhydrous ethanol, and dried at 60 °C to obtain the final product (PCC).

[0038] Morphological detection: The final product obtained in Example 1 was subjected to X-ray photoelectron detection, atomic force microscopy, and transmission electron microscopy. The results are as follows: Figures 1-4 As shown, we can see the following points: (1) The material has an irregular layered structure with a uniform height distribution of less than 1 nm. (2) Palladium, copper, and cobalt are uniformly distributed in space on the nanosheet. (3) The final product is mainly composed of palladium, copper, and cobalt.

[0039] Biosafety and biocompatibility testing: PCC was dissolved in phosphate-buffered saline (PBS) to prepare solutions with concentrations of 0, 5 mg / kg, 10 mg / kg, and 20 mg / kg. Healthy mice were randomly divided into groups of 5, and each mouse was injected with 100 μL via the tail vein.

[0040] The body weight changes of the four groups of mice were monitored over 14 days, and the results are as follows: Figure 5 As shown in the table, no significant changes in body weight or abnormal behavior were observed in any group of mice. Blood biochemical analyses were then performed on each group of mice, and the results are shown in Tables 1 and 2. Table 1 shows the data for liver and kidney function-related markers, and Table 2 shows the data for routine blood tests.

[0041] Table 1. Detection data of liver and kidney function-related biomarkers

[0042] Table 2 Blood routine index data

[0043] The results in Tables 1 and 2 show that PCC has almost no effect on liver and kidney function markers and has good blood compatibility.

[0044] Major organs (heart, liver, spleen, lung, and kidney) of mice were collected and stained with hematoxylin and eosin (H&E). The results are as follows: Figure 6 As shown. For Figure 6 Histopathological analysis showed that no significant damage or inflammatory response was observed in any organ after intravenous injection of PCC, further verifying that PCC has ideal tissue compatibility.

[0045] Therefore, it can be demonstrated that PCC has good biosafety and biocompatibility.

[0046] Tumor treatment efficacy testing: Twenty tumor-bearing mice with subcutaneous xenografts of 4T1 breast cancer were randomly divided into four groups of five mice each. The first group received a PBS injection (PBS group). The second group also received a PBS injection, followed by ultrasound of the tumor (US group). The third group received a 20 mg / kg PCC injection (PCC group). The fourth group received a 20 mg / kg PCC injection, followed by ultrasound of the tumor site (PCC+US group). All four groups received the treatment on days 1, 4, 7, and 10, with an injection volume of 100 μL. The ultrasound treatment for the second and fourth groups was low-frequency ultrasound irradiation at a frequency of 1.0 MHz and a power density of 1.2 W / cm². 2 Duty cycle 50%, irradiation time 3 min.

[0047] During the 14-day experimental period following injection, the body weight and tumor volume of mice in each group were closely monitored every other day, and the results are as follows: Figure 7 , Figure 8As shown in the figure. Results showed that the body weight of mice in all groups did not fluctuate significantly during treatment. Tumor growth curves indicated that, compared with the control group and the ultrasound-only group, the PCC-only group showed only a moderate inhibitory effect on tumor progression, while the PCC-ultrasound combined group showed the most significant tumor-inhibiting effect. At the experimental endpoint, the tumor volume in the PCC-ultrasound combined group was significantly smaller than that in other groups, with a tumor inhibition rate as high as 64%, the highest among all groups, confirming that ultrasound activation can significantly enhance the in vivo antitumor efficacy of PCC.

[0048] Simultaneously, PCC combined with ultrasound therapy can inhibit the metastasis of triple-negative breast cancer, with results as follows: Figure 9 As shown in the figure, the lung tissue weight of mice in the PCC combined with ultrasound group was significantly lower than that of other groups, and no obvious metastatic nodules were found in the lung tissue.

[0049] The above description of the embodiments is intended to enable those skilled in the art to understand and apply this application. It will be apparent to those skilled in the art that various modifications can be easily made to these embodiments, and the general principles described herein can be applied to other embodiments without creative effort. Therefore, this application is not limited to the embodiments described herein, and any improvements and modifications made by those skilled in the art based on the disclosure of this application without departing from the scope and spirit of this application are within the scope of this application.

Claims

1. A palladium-copper-cobalt nanomaterial, characterized in that, The nanomaterial contains palladium, copper and cobalt metals, and the molar ratio of palladium, copper and cobalt metals is 1:(7~8):(15~17). The nanomaterial has a layered structure.

2. The palladium-copper-cobalt nanomaterial as described in claim 1, characterized in that, The particle size of the nanomaterial is 300~320nm.

3. The palladium-copper-cobalt nanomaterial as described in claim 1, characterized in that, The thickness of each layer of the nanomaterial is 0.6~0.9 nm.

4. The palladium-copper-cobalt nanomaterial as described in claim 1, characterized in that, The nanomaterial is modified with PVP by electrostatic adsorption, wherein the mass ratio of the nanomaterial to PVP is 1:15~30.

5. A method for preparing palladium-copper-cobalt nanomaterials as described in any one of claims 1 to 3, characterized in that, The preparation method includes the following steps: The palladium source, copper source, cobalt source, hexacarbonyl tungsten and reducing agent are dissolved in a solvent, and then subjected to ultrasonic treatment, heating, cooling, centrifugation, washing and drying in sequence to obtain the nanomaterial.

6. The method for preparing palladium-copper-cobalt nanomaterials as described in claim 5, characterized in that, The palladium source is palladium diacetylacetonate, the copper source is copper diacetylacetonate, the cobalt source is cobalt diacetylacetonate, the reducing agent is ascorbic acid, and the molar ratio of the palladium source, copper source, cobalt source, tungsten hexacarbonyl and reducing agent is (1.0~1.1):(1.2~1.3):1.0:(5.4~5.6):(7.2~7.4); the solvent is a mixed solution of N,N-dimethylformamide and acetic acid, and the volume ratio of N,N-dimethylformamide to acetic acid is (3~5):

1.

7. The method for preparing palladium-copper-cobalt nanomaterials as described in claim 5, characterized in that, The preparation method includes at least one of the following features: 1) The ultrasonic treatment is performed at room temperature for 0.5 to 2 hours; 2) The heating is carried out in an oil bath at a temperature of 60-100℃ for 8-15 hours. 3) The cooling is natural cooling, cooling to room temperature; 4) The centrifugation speed is 6000~10000 rpm; the centrifugation time is 5~20 min; 5) The washing process uses ethanol and involves at least 5 washes; 6) The drying temperature is 40~60℃ and the drying time is 6~8h.

8. The method for preparing palladium-copper-cobalt nanomaterials as described in claim 5, characterized in that, The nanomaterials are dissolved, modified, washed twice, and dried to obtain nanomaterials modified with PVP.

9. The method for preparing palladium-copper-cobalt nanomaterials as described in claim 8, characterized in that, The preparation method includes at least one of the following features: a) The dissolution is achieved by deionizing the product solution in water; b) The modification involves adding PVP and then performing magnetic stirring, wherein the mass ratio of the nanomaterial to PVP is 1:(15~30), the stirring rate of the magnetic stirring is 100~200 rpm, and the stirring time is 20~30 h; c) The secondary washing is performed by alternating washing with deionized water and anhydrous ethanol, and the number of washing cycles is not less than 3. d) The drying temperature is 40~60℃ and the drying time is 6~8h.

10. An application of the palladium-copper-cobalt nanomaterial as described in any one of claims 1 to 4, characterized in that, The nanomaterials are used to make injections for treating breast cancer.

11. The application of the palladium-copper-cobalt nanomaterial as described in claim 10, characterized in that, The active ingredient of the injection is the nanomaterial, the solvent of the injection is phosphate buffer, and the concentration of the nanomaterial is 15~25 mg / kg.

12. The application of the palladium-copper-cobalt nanomaterial as described in claim 10, characterized in that, After the injection, ultrasound manipulation is performed on the tumor site.

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