New uses for hydrated copper ions

Abstract

This invention belongs to the field of biomedical technology, specifically disclosing a novel application of hydrated copper ions. Hydrated copper ions exhibit strong absorption in the 808 nm near-infrared range, demonstrating excellent photothermal effects. Utilizing the ultra-small structure and high photothermal conversion performance of hydrated copper ions, they can kill tumor cells, achieving a therapeutic purpose. A mouse model constructed using this invention demonstrates the effectiveness of subcutaneous injection of Cu in mice with prostate cancer. 2+ The solution was tested using a wavelength of 808 nm and a power of 1.5 W·cm⁻¹. ‑2 Infrared laser irradiation of the tumor treatment area for 10 minutes resulted in nearly 100% inhibition of prostate cancer cells in mice after treatment. During the 14-day treatment period, the mice in the treatment group did not experience any significant weight loss and had a high survival rate, demonstrating excellent photothermal therapeutic effects on tumor cells and good biocompatibility in mice.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to novel uses of hydrated copper ions. Background Technology

[0002] Cancer is a major disease that seriously threatens human health and has become one of the most destructive diseases in the world, characterized by high incidence and mortality rates. The prevention and control of malignant tumors is extremely challenging, and improving the survival rate of cancer patients is a pressing problem that humanity needs to overcome. Chemotherapy (CT) is considered the most commonly used treatment method. However, chemotherapy has many limitations, such as low drug loading, high dosage, and unavoidable carrier toxicity, which limits its widespread use. Photodynamic therapy (PDT), photothermal therapy (PTT), and chemodynamic therapy (CDT), as emerging tumor treatment methods, have, to some extent, compensated for the shortcomings of traditional treatments due to their advantages of low invasiveness, high selectivity, and repeatability. Among them, photothermal therapy (PTT) is an important approach, known as a "green therapy" due to its advantages of rapid targeted killing, minimal invasiveness, and few toxic side effects, and has great potential in the field of tumor treatment. The main working principle of PTT is to utilize photothermal conversion nanomaterials (PTA) with strong absorption in the near-infrared region to absorb light energy and then convert it into heat energy through non-radiative relaxation, thereby raising the temperature of local tumor tissue and effectively killing tumor cells. In the photothermal conversion process (PTT), photothermal conversion nanomaterials play a crucial role. Currently, most photothermal conversion materials used clinically are based on inorganic materials (including noble metals, transition metal chalcogenides, and carbon-based nanomaterials), organic materials, and organic-inorganic composite materials. However, these photothermal material systems all face problems in clinical application, such as metabolic difficulties, multiple components and complex mechanisms of action, and difficulty in controlling side effects.

[0003] Therefore, photothermal materials with high photothermal conversion efficiency, water solubility, easy metabolism, and single composition are developed to solve the above problems. Summary of the Invention

[0004] The purpose of this invention is to address the aforementioned shortcomings of existing technologies by providing the application of hydrated copper ions in the preparation of antitumor drugs. Tumor cells have a heat resistance temperature of 39.5℃ to 42.5℃. When the temperature reaches above 42.5℃, the tumor cells will be in a state of shock. The principle of tumor photothermal therapy is to use physical methods to heat tissue to a temperature that can kill tumor cells. This invention utilizes the ultra-small structure and high photothermal conversion performance of hydrated copper ions to kill tumor cells and achieve the purpose of treatment.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] The first objective of this invention is to provide the application of hydrated copper ions in the preparation of antitumor drugs, wherein the hydrated copper ions convert light energy into heat energy to kill tumor cells under infrared laser irradiation.

[0007] Furthermore, the method for preparing the hydrated copper ions is to dissolve a soluble copper salt in water.

[0008] Furthermore, the soluble copper salt is any one of copper chloride, copper sulfate, and copper nitrate.

[0009] Furthermore, the tumor includes prostate cancer cells.

[0010] Furthermore, the irradiation conditions for the infrared laser are: laser wavelength of 808 nm and power of 1.5 W·cm⁻¹. -2 Irradiation time: 10-15 minutes.

[0011] Furthermore, the hydrated copper ions, in conjunction with infrared laser irradiation, have an inhibitory effect on the proliferation of prostate cancer cells.

[0012] A second object of the present invention is to provide a pharmaceutical composition comprising the above-described hydrated copper ions.

[0013] A third objective of this invention is to provide an anti-prostate cancer drug comprising the aforementioned hydrated copper ions.

[0014] Furthermore, the hydrated copper ions are the only effective active ingredient.

[0015] Furthermore, the anti-prostate cancer drug also includes pharmaceutically acceptable excipients.

[0016] The term "excipient" broadly refers to any component other than the active ingredient. Excipients can be inert, inactive, and / or substances without medicinal activity.

[0017] Excipients can serve various purposes, such as acting as carriers, solvents, diluents, tablet additives, and / or enhancing the delivery and / or absorption of active substances. Excipients are not limited herein and are selected based on the formulation process, and include, but are not limited to: solvents, diluents, buffers, preservatives, isotonic agents, pH adjusters, surfactants, adjuvants, ionic strength enhancers, chelating agents, stabilizers, or any combination thereof.

[0018] Furthermore, the dosage form of the anti-prostate cancer drug includes any pharmaceutically acceptable dosage form.

[0019] The medicaments of this invention can be formulated into any dosage form known in the medical field, such as capsules, granules, oral solutions, tablets, or injections. Preferred dosage forms depend on the intended route of administration and preventative / therapeutic use. For example, the medicament may be a liquid formulation, i.e., an aqueous formulation containing water. Liquid formulations may be solutions or suspensions. Aqueous formulations typically contain at least 50% w / w water or at least 60%, 70%, 80%, or even at least 90% w / w water. Alternatively, the medicament may be a solid formulation, such as a lyophilized or spray-dried composition, which can be used as is, or a physician or patient may add solvents and / or diluents to it immediately before use.

[0020] The pH of aqueous formulations can be any value between pH 3 and pH 10, for example, about 7.0 to about 9.5; or about 3.0 to about 7.0.

[0021] In accordance with conventional formulation processes in the art, in specific embodiments, for example, an injectable formulation may be prepared by adding a necessary dose of hydrated copper ions to a suitable solvent, and optionally, simultaneously incorporating other improving components (including, but not limited to, solvents, diluents, buffers, preservatives, isotonic agents, pH adjusters, surfactants, adjuvants, ionic strength enhancers, chelating agents, and stabilizers, or any combination thereof), followed by sterilization by filtration. Alternatively, a lyophilized formulation may be prepared by incorporating a necessary dose of the hydrated copper ions of the present invention to a suitable solvent, and optionally, simultaneously incorporating other desired components (solvents, diluents, buffers, preservatives, isotonic agents, pH adjusters, surfactants, adjuvants, ionic strength enhancers, chelating agents, stabilizers, or any combination thereof), followed by sterilization by filtration, lyophilization, or direct freezing.

[0022] Compared with the prior art, the beneficial effects of the present invention are:

[0023] (1) The application of hydrated copper ions in the preparation of antitumor drugs provided by the present invention: hydrated copper ions have strong absorption at 808 nm near-infrared light, showing excellent photothermal effect. By utilizing the ultra-small structure and high photothermal conversion performance of hydrated copper ions, tumor cells are killed to achieve the purpose of treatment.

[0024] (2) The Cu provided by the present invention 2+ The solution was tested at a wavelength of 808 nm and a power of 1.5 W·cm. -2 It exhibits a photothermal conversion efficiency of 72.41% under infrared laser irradiation, making it a novel photothermal conversion material.

[0025] (3) Cu provided by the present invention 2+ The solution exhibits excellent biocompatibility in mice.

[0026] (4) This invention demonstrates, through the construction of a mouse model, that subcutaneous injection of Cu in mice with prostate cancer 2+ Solution (200 μg / mL, 100 μL), with a wavelength of 808 nm and a power of 1.5 W·cm⁻¹. -2 Infrared laser irradiation of the tumor treatment area for 10 minutes resulted in nearly 100% inhibition of prostate cancer cells in mice after treatment. During the 14-day treatment period, the mice in the treatment group did not experience any significant weight loss and had a high survival rate. Attached Figure Description

[0027] Figure 1 Different concentrations of Cu in this invention 2+ EXFAS (Extra-Factor Spectroscopy) plot of the solution; from the figure, it can be seen that Cu 2+ Cu-Cu coordinate aggregation occurs in aqueous solution;

[0028] Figure 2 Different concentrations of Cu in this invention 2+ (a) UV-vis-NIR absorption spectrum of the solution; (b) Absorption fitting line at 808 nm; From the figure, it can be seen that Cu 2+ The solution exhibits strong absorption in the near-infrared region at 808 nm, and this absorption is concentration-dependent.

[0029] Figure 3 Different concentrations of Cu in this invention 2+ The solution was tested at a wavelength of 808 nm and a power of 1.5 W·cm⁻¹. -2 The graph shows the temperature change after 10 minutes of laser irradiation; it can be seen from the graph that the concentration of Cu is 0.05 mol / L. 2+ The solution was subjected to an 808 nm laser at a wavelength of 1.5 W·cm⁻¹. -2 After 10 minutes of irradiation, the temperature can rise to 84°C, a temperature change sufficient to kill tumor cells;

[0030] Figure 4 The Cu concentration of this invention is 0.03 mol / L. 2+ The solution was tested at a wavelength of 808 nm and a power of 1.5 W·cm⁻¹. -2 (a) Heating and cooling curves under laser irradiation; (b) Photothermal conversion efficiency graph; Calculations are performed using the heating and cooling data in graph (a) to obtain Cu 2+ The solution was tested at a wavelength of 808 nm and a power of 1.5 W·cm⁻¹. -2 The photothermal conversion efficiency under laser irradiation is 72.41%;

[0031] Figure 5 The human-derived cells (HK-2 cells) and mouse prostate cancer cells (RM-1 cells) of the present invention were subjected to different concentrations and Cu.2+ Cell viability was measured after 24 hours of culture in solution, and cell viability was detected using the standard MTT assay. The figure shows that Cu... 2 + The solution exhibited an IC50 of 12-16 μg / mL against mouse prostate cancer cells (RM-1 cells), while simultaneously maintaining a high survival rate of 85% for human-derived cells (HK-2 cells), indicating that Cu... 2+ The solution exhibits good biocompatibility;

[0032] Figure 6 The PBS group and Cu of the present invention 2+ The curve showing the change in body weight of mice in the photothermal therapy group over time. As can be seen from the figure, during the 14-day treatment period, Cu... 2+ Mice in the photothermal therapy group did not show any significant weight loss;

[0033] Figure 7 The PBS group and Cu of the present invention 2+ Survival curves of mice in the photothermal therapy group; as shown in the figure, Cu 2+ The survival rate of mice in the photothermal therapy group was significantly higher than that in the PBS group;

[0034] Figure 8 The PBS group and Cu of the present invention 2+ Growth curves of tumor volume after photothermal therapy; the graph shows that the tumors in the PBS group continued to grow, while those in the Cu group... 2+ The photothermal therapy group achieved a tumor inhibition rate of nearly 100%.

[0035] Figure 9 The PBS group and Cu of the present invention 2+ Photographs of tumors in the photothermal therapy group on day 14 after treatment. The images clearly show that, compared to the PBS treatment group, Cu... 2+ The tumors in the photothermal therapy group were significantly suppressed. Detailed Implementation

[0036] To make the objectives, technical solutions, and advantages of the present invention clearer, the specific embodiments of the present invention will be described in further detail below with reference to specific examples and accompanying drawings.

[0037] Unless otherwise specified, the experimental methods used in the embodiments are conventional methods, and the materials and reagents used are commercially available unless otherwise specified.

[0038] In this invention, "hydrated copper ions" is equivalent to "Cu". 2+"Solution" refers to the fact that because water is a strongly polar molecule, it can dissolve copper salts as a solvent, and can also combine with dissolved copper ions to form clusters. This process is called ion hydration, and the resulting ion-hydrated clusters are called "hydrated copper ions".

[0039] Example 1

[0040] Preparation of hydrated copper ions

[0041] Weigh 0.1705 g of CuCl2·2H2O into a centrifuge tube, add 10 mL of deionized water, shake well, and prepare a CuCl2 solution with a concentration of 0.1 mol / L. 2+ Solution, for backup.

[0042] Example 2

[0043] Cu 2+ UV absorption of solution

[0044] The Cu prepared above 2+ Solutions were prepared into a series of samples with concentrations (0.01, 0.02, 0.03, and 0.05 mol / L), and the spectral absorption in the 400-1300 nm range was measured using a UV-Vis spectrophotometer. Cu 2+ The solution exhibits strong absorption at 808 nm near-infrared light, and this absorption is concentration-dependent.

[0045] When preparing hydrated copper ion solutions using copper nitrate or copper sulfate, it was also found that in Cu... 2+ The solution exhibits strong absorption at 808 nm near-infrared light, and this absorption is concentration-dependent.

[0046] Example 3

[0047] Cu 2+ Photothermal conversion efficiency of solution

[0048] Use a pipette to draw 1 mL of 0.03 mol / L Cu solution. 2+ The solution was placed in a 1.5 mL centrifuge tube, and a centrifuge with a wavelength of 808 nm and a power of 1.5 W·cm⁻¹ was used. -2 An infrared laser was placed directly above the centrifuge tube. After irradiating the solution for 10 minutes, the laser was immediately turned off. The solution was allowed to cool to room temperature (25°C), and the Cu content was recorded using an infrared thermal imager. 2+ The temperature change of the solution during laser irradiation and after irradiation was recorded every 30 seconds to obtain a temperature rise and fall curve. Using the formula for photothermal conversion efficiency, the Cu content can be calculated. 2+ Photothermal conversion efficiency of the solution.

[0049] Example 4

[0050] Cu2+ Solution cytotoxicity assay

[0051] 1) Human cells (HK-2 cells) and mouse prostate cancer cells (RM-1 cells) were seeded at a density of 8000 cells / well into 96-well plates (with spaced-apart plates and PBS added to the spacers to insulate against heat) and cultured for 24 hours.

[0052] 2) Prepare Cu solutions with different concentration gradients 2+ Solutions (0, 4, 8, 12, 16, 20 μg / mL): Take 0, 16, 32, 48, 64, and 80 μL of Cu. 2+ The solution (stock solution concentration of 50 μg / ml) was mixed thoroughly in 200, 184, 168, 136, and 120 μl of complete culture medium.

[0053] 3) Then take 200 μl of the prepared Cu solutions with different concentration gradients as described above. 2+ Cells were treated with solutions (0, 4, 8, 12, 16, 20 μg / mL) for 30 min, washed twice with PBS, incubated for 24 h, and cell viability was detected by the standard MTT assay.

[0054] Example 5

[0055] Cu 2+ In vivo animal antitumor experiments with solution

[0056] Black mice were used as the mouse species, and prostate cancer cells (RM-1) were used as the cancer cell model. First, RM-1 cells were cultured for 2-3 passages to reach a sufficient population, ideally 1.0 × 10⁶ cells per mouse. 5 100 μL of tumor cells were subcutaneously injected into mice. Once the tumor cells stabilized and reached a size of 100 mm in the mice, the injection was continued. 3 Subsequently, the mice were divided into an experimental group and a control group. The experimental group received intratumoral injection of Cu. 2+ Solution (200 μg / mL, 100 μL), with a wavelength of 808 nm and a power of 1.5 W·cm⁻¹. -2 The tumor treatment area was irradiated for 10 minutes; the control group was injected with PBS solution only. After the treatment, the tumor treatment status of mice was observed for 14 days (tumor inhibition rate statistics for each group; survival statistics (N=4); representative tumor photos of mice in each group).

[0057] For any points not covered above, existing technologies shall apply.

[0058] Although specific embodiments of the present invention have been described in detail by way of examples, those skilled in the art should understand that the above examples are for illustrative purposes only and are not intended to limit the scope of the invention. Those skilled in the art can make various modifications or additions to the described specific embodiments or use similar methods to replace them, without departing from the direction of the invention or exceeding the scope defined by the appended claims. Those skilled in the art should understand that any modifications, equivalent substitutions, improvements, etc., made to the above embodiments based on the technical essence of the present invention should be included within the protection scope of the present invention.

Claims

1. The application of hydrated copper ions in the preparation of antitumor drugs, characterized in that, The hydrated copper ions convert light energy into heat energy to kill tumor cells under infrared laser irradiation. The method for preparing the hydrated copper ions is to dissolve a soluble copper salt in water. The soluble copper salt is any one of copper chloride, copper sulfate, and copper nitrate; The tumor in question is prostate cancer; The irradiation conditions for the infrared laser are: laser wavelength of 808 nm and power of 1.5 W·cm. -2 The irradiation time is 10-15 minutes; the hydrated copper ions are the only effective active ingredient.

2. The application as described in claim 1, characterized in that, The hydrated copper ions, in conjunction with infrared laser irradiation, have an inhibitory effect on the proliferation of prostate cancer cells.

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