Indocyanine green stabilized CuET nanoparticles, nanoparticle dispersions, their preparation methods and applications
By using an indocyanine green-stabilized CuET nanoparticle dispersion, the problems of poor water solubility of CuET and limited photodynamic activity of indocyanine green were solved, enabling intravenous administration of CuET and enhanced antitumor therapeutic effects.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-09-28
- Publication Date
- 2026-04-03
AI Technical Summary
CuET has poor water solubility, making it difficult to prepare intravenous formulations in a simple and safe manner. Furthermore, the photodynamic effect of indocyanine green is limited by the hypoxic environment of tumors.
Indocyanine green-stabilized CuET nanoparticles are used, and CuET nanocrystals are combined through hydrophobic interactions to form a nanoparticle dispersion. The photodynamic effect of indocyanine green is then utilized to enhance anti-tumor therapy.
It improved the dispersibility of CuET in water and its serum stability, enhanced its killing effect on tumor cells, and improved the photodynamic antitumor effect of indocyanine green.
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of nanomaterials technology, and more specifically, relates to indocyanine green stabilized CuET nanoparticles, nanoparticle dispersions, their preparation methods, and applications. Background Technology
[0002] In recent years, disulfiram, a drug used to treat alcoholism, has been found to have anti-tumor effects. Studies have shown that the mechanism by which disulfiram exerts its anti-cancer function is mainly due to the reaction of its metabolite DDTC salt ions with Cu(II) ions to generate CuET. CuET can exert its anti-tumor effect by interfering with normal mitochondrial function or protein degradation. However, the bioavailability of orally administered disulfiram is low, and very little reaches the tumor site. Furthermore, the amount of Cu ions in the body is insufficient to form large amounts of CuET. In addition, CuET has poor water solubility, making intravenous administration impossible. These factors limit the application of CuET in cancer treatment.
[0003] To address the issue of water solubility, researchers have loaded CuET onto carriers such as fucoidan, liposomes, polyvinylpyrrolidone, and melanin to improve its dispersibility in water, facilitating intravenous administration. However, these methods suffer from one or more of the following problems: 1. The carrier or excipient components are loaded without clinical approval; 2. The preparation process is complex and costly; 3. Impurity removal is difficult and may introduce toxic organic solvents.
[0004] Indocyanine green (ICB) is a near-infrared fluorescent dye approved by the U.S. Food and Drug Administration (FDA) and widely used in medical imaging diagnostics. It has been used clinically for over 50 years and has a good safety profile. ICB also possesses photodynamic effects and has been widely applied in clinical and preclinical research for the diagnosis and treatment of tumors in recent years. However, the hypoxic microenvironment of solid tumors limits the full potential of ICB's photodynamic effects. Summary of the Invention
[0005] To address the shortcomings of existing technologies, this invention aims to provide indocyanine green-stabilized CuET nanoparticles, nanoparticle dispersions, their preparation methods, and applications, thereby solving the technical problem of CuET's poor water solubility, which makes it difficult to prepare intravenous formulations in a simple and safe manner. Furthermore, this invention utilizes CuET to interfere with mitochondrial function, inhibiting cellular respiration and alleviating tumor site hypoxia, thus addressing the limitation of indocyanine green photodynamic effects by tumor hypoxia.
[0006] To achieve the above objectives, the present invention provides indocyanine green stabilized CuET nanoparticles, which are nanoparticles formed by stabilizing CuET nanocrystals with indocyanine green, wherein indocyanine green binds to and stabilizes CuET nanocrystals through hydrophobic interactions to form the indocyanine green stabilized CuET nanoparticles.
[0007] Preferably, the particle size of the indocyanine green stabilized CuET nanoparticles is 10–400 nanometers.
[0008] According to another aspect of the invention, an indocyanine green-stabilized CuET nanoparticle dispersion is provided, comprising the indocyanine green-stabilized CuET nanoparticles, wherein the indocyanine green binds to and stabilizes the CuET nanocrystals through hydrophobic interactions, and is dispersed in water to form the dispersion.
[0009] Preferably, the concentration of the indocyanine green-stabilized CuET nanoparticles in the dispersion is 0.1–100 mg / mL.
[0010] According to another aspect of the present invention, a method for preparing the indocyanine green-stabilized CuET nanoparticle dispersion is provided, comprising the following steps: mixing Cu(II) ion salt solution and DDTC salt solution in an indocyanine green aqueous solution to allow them to react and obtain an indocyanine green-stabilized CuET nanoparticle dispersion.
[0011] Preferably, the molar ratio of copper(II) ions to indocyanine green in the Cu(II) ion salt solution is 5–40:1; the molar ratio of DDTC ions to Cu(II) ions is 1.5–2.2:1; and the concentration of Cu(II) ions in the dispersion is 0.35–2.78 mM.
[0012] Preferably, the Cu(II) ion salt is copper nitrate, copper sulfate, or copper chloride; the DDTC salt solution is an aqueous solution of sodium DDTC or potassium DDTC.
[0013] Preferably, the Cu(II) ion salt solution is added dropwise to the DDTC salt solution and the aqueous solution of indocyanine green, and the mixture is stirred during the dropwise addition to allow the reaction to occur, thereby obtaining an indocyanine green-stabilized CuET nanoparticle dispersion.
[0014] Preferably, the reaction temperature is 30–70°C, more preferably 40–70°C, and even more preferably 50–60°C.
[0015] According to another aspect of the invention, the use of the indocyanine green stabilized CuET nanoparticles and / or the indocyanine green stabilized CuET nanoparticle dispersion described herein is provided in the preparation of medicaments for the prevention and / or treatment of tumors.
[0016] According to another aspect of the invention, an anticancer drug is provided, comprising the indocyanine green stabilized CuET nanoparticles and / or the indocyanine green stabilized CuET nanoparticle dispersion.
[0017] In summary, compared with the prior art, the above-described technical solutions conceived by this invention have the following advantages:
[0018] Beneficial effects:
[0019] (1) This invention provides an indocyanine green-stabilized CuET nanoparticle, which is a nanoparticle formed by stabilizing CuET nanocrystals with indocyanine green. Indocyanine green binds to the CuET nanocrystals through hydrophobic interactions, forming the indocyanine green-stabilized CuET nanoparticle. The hydrophobic interactions between indocyanine green and CuET allow the CuET nanocrystals to be stably dispersed in water, forming an indocyanine green-stabilized CuET nanoparticle dispersion. The indocyanine green-stabilized CuET nanoparticles provided by this invention can improve the dispersibility of CuET in water, enhance the killing effect of CuET on tumor cells, alleviate tumor hypoxia, and enhance the photodynamic antitumor effect of indocyanine green in vivo.
[0020] (2) This invention uses indocyanine green, which is widely used in clinical practice, as a stabilizer. Cu(II) ion salt solution and DDTC salt solution are mixed in an indocyanine green aqueous solution to prepare indocyanine green-stabilized CuET nanoparticles. Indocyanine green-stabilized CuET nanoparticles can be prepared in aqueous solution using a simple method. The materials used in this preparation process are all used in clinical or clinical trials, and have great potential for clinical application. Attached Figure Description
[0021] Figure 1 Photographs of the samples prepared in Example 1 and Comparative Example 1;
[0022] Figure 2 The results show the dynamic light scattering particle size distribution of samples 1–6;
[0023] Figure 3 The TEM results are for sample 1.
[0024] Figure 4 The XRD results are for sample 1;
[0025] Figure 5 The mass spectrometry results are for sample 1;
[0026] Figure 6 The results show the serum stability of sample 1;
[0027] Figure 7 The results show the mitochondrial membrane potential of 4T1 cells after treatment with sample 1;
[0028] Figure 8 The results show the oxygen consumption of 4T1 cells after treatment with sample 1;
[0029] Figure 9The hypoxia status of 4T1 cells after treatment with sample 1;
[0030] Figure 10 The assessment results of the ability of Sample 1 to generate singlet oxygen;
[0031] Figure 11 The efficacy results of combined photodynamic therapy for subcutaneous tumors of 4T1 breast cancer in Sample 1;
[0032] Figure 12 The effect of reaction temperature on the formation of indocyanine green-stabilized CuET nanoparticle dispersions. Detailed Implementation
[0033] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.
[0034] To address the technical problem that the poor water solubility of CuET in existing technologies limits its application in cancer treatment, this invention provides indocyanine green-stabilized CuET nanoparticles. These nanoparticles are formed by indocyanine green-stabilized CuET nanocrystals, wherein indocyanine green is bonded to the CuET nanocrystals through hydrophobic interactions to form the indocyanine green-stabilized CuET nanoparticles.
[0035] The present invention also provides an indocyanine green-stabilized CuET nanoparticle dispersion, comprising the indocyanine green-stabilized CuET nanoparticles as described above. Indocyanine green binds to and stabilizes CuET through hydrophobic interactions, enabling the CuET nanocrystals to be stably dispersed in water to form an indocyanine green-stabilized CuET nanoparticle dispersion.
[0036] In this invention, indocyanine green is used as a stabilizer, possibly because the hydrophobic interaction between indocyanine green and CuET prevents the aggregation and precipitation of CuET nanocrystals, thereby improving the dispersibility of CuET nanocrystals in water and their stability in serum. Indocyanine green itself is a small molecule compound, and its role as a stabilizer for CuET nanocrystals is not readily apparent.
[0037] In some embodiments, the average particle size of the indocyanine green-stabilized CuET nanoparticles is 10–400 nm, preferably 10–200 nm, and more preferably 40–100 nm. The concentration of the indocyanine green-stabilized CuET nanoparticles in the dispersion is 0.1–100 mg / mL, preferably 0.1–10 mg / mL, and the PDI (aggregation index, a parameter describing the uniformity of particle size distribution) of the indocyanine green-stabilized CuET nanoparticles in the dispersion is less than or equal to 0.4, preferably less than or equal to 0.2.
[0038] This invention provides a method for preparing the indocyanine green-stabilized CuET nanoparticle dispersion, comprising the following steps: a Cu(II) ion salt solution is added dropwise to the DDTC salt solution and the indocyanine green aqueous solution, while the mixture is stirred at a constant temperature during the dropwise addition, to obtain the indocyanine green-stabilized CuET nanoparticle dispersion. In this invention, adding the Cu(II) ion salt solution dropwise to the DDTC salt solution and the indocyanine green aqueous solution indicates that an aqueous solution of indocyanine green and DDTC salt is prepared first, and then the Cu(II) ion salt solution is added dropwise to the indocyanine green and DDTC salt aqueous solution. However, it is not advisable to directly mix the Cu(II) ion salt solution and the DDTC salt solution beforehand, nor is it advisable to pre-mix the Cu(II) ion salt solution with the indocyanine green aqueous solution; otherwise, it will be difficult to obtain an indocyanine green-stabilized CuET nanoparticle dispersion with uniform particle size and high repeatability.
[0039] Sodium DDTC, also known as sodium diethyldithiocarbamate or sodium diethylcarbamate, has the CAS Registry Number 148-18-5 and the chemical formula C5H. 10 NNaS2.
[0040] In some embodiments, the molar ratio of copper(II) ions to indocyanine green in the Cu(II) ion salt solution is 5–40:1, preferably 5–22.5:1; the Cu(II) ion salt can be copper nitrate, copper sulfate, or copper chloride, etc.; the concentration of Cu(II) ions in the dispersion is 0.35–2.78 mM; the molar ratio of DDTC ions to Cu(II) ions is 1.5–2.2:1; the DDTC salt solution can be an aqueous solution of sodium DDTC or potassium DDTC, etc.
[0041] In a preferred embodiment, the Cu(II) ion salt solution is added dropwise to the DDTC salt solution and the aqueous solution of indocyanine green, and stirring is performed simultaneously to obtain an indocyanine green-stabilized CuET nanoparticle dispersion; the stirring includes, but is not limited to, magnetic stirring or mechanical stirring.
[0042] In some embodiments, the Cu(II) ion salt solution is added dropwise to the DDTC salt solution and the indocyanine green aqueous solution at a rate of 0.01–5 mL / s, preferably 0.01–0.1 mL / s. The stirring method is magnetic stirring or mechanical stirring at a speed of 10–2600 rpm, more preferably 200–2000 rpm.
[0043] The preparation of indocyanine green-stabilized CuET nanoparticles using the above method can be carried out over a wide temperature range, such as between 30 and 70°C, preferably between 40 and 70°C, and even more preferably between 50 and 60°C.
[0044] This invention obtains an indocyanine green-stabilized CuET nanoparticle dispersion by adding the Cu(II) ion salt solution dropwise to the DDTC salt solution and the indocyanine green aqueous solution, thereby improving the dispersibility of CuET in water and its stability in serum. Experiments have shown that the indocyanine green-stabilized CuET nanoparticle dispersion of this invention can be used to prepare drugs for treating tumors, including but not limited to breast cancer and liver cancer, and can be administered via intravenous injection, intratumoral injection, or intraperitoneal injection.
[0045] The indocyanine green-stabilized CuET nanoparticles provided by this invention can improve the water solubility and serum stability of CuET, facilitating intravenous administration of CuET. Furthermore, the indocyanine green-stabilized CuET nanoparticles provided by this invention can disrupt mitochondrial function in tumor cells, inhibit tumor cell respiration, improve hypoxia at the tumor site, and enhance the photodynamic antitumor therapeutic effect mediated by indocyanine green.
[0046] The present invention also provides an anticancer drug comprising the indocyanine green stabilized CuET nanoparticles and / or the indocyanine green stabilized CuET nanoparticle dispersion, and may further comprise pharmaceutically acceptable additives.
[0047] The following is an example:
[0048] Example 1
[0049] This embodiment provides indocyanine green-stabilized CuET nanoparticles, using CuCl2 and DDTC sodium salt aqueous solution as the reaction ionic solution. The molar ratio of Cu(II) to indocyanine green in the nanoparticles is 10:1. The preparation process is as follows:
[0050] (1) Dissolve 0.425 mg of indocyanine green in 6 mL of water, then add 1 mL of DDTC sodium aqueous solution with a concentration of 2.5 mg / mL, and in an oil bath at 50 °C to obtain mixed solution A;
[0051] (2) While stirring, 1 mL of a 0.94 mg / mL CuCl2·2H2O aqueous solution was added dropwise to the obtained mixed solution A over a period of 60 s. The mixture was stirred magnetically at 1200 rpm at a temperature of 50 °C. Once the addition was complete, an indocyanine green-stabilized CuET nanoparticle dispersion was obtained. The sample was purified and concentrated by ultrafiltration (100 kDa molecular weight cutoff). This sample was named Sample 1.
[0052] Comparative Example 1
[0053] The preparation of CuET includes the following steps:
[0054] Add 1 mL of 0.94 mg / mL CuCl2·2H2O aqueous solution dropwise to 1 mL of 2.5 mg / mL DDTC sodium aqueous solution. After the addition is complete, centrifuge, wash, and resuspend in deionized water to obtain the CuET sample.
[0055] Figure 1 The images show the CuET samples prepared for this comparative example and Sample 1 prepared in Example 1 after being placed for 12 hours. It can be seen that the CuET in the CuET sample prepared for this comparative example (left side) is obviously agglomerated and precipitated, while Sample 1 in Example 1 (right side) is a stable dispersion.
[0056] Example 2
[0057] This embodiment provides indocyanine green-stabilized CuET nanoparticles, and the preparation process is as follows:
[0058] Using CuCl2 and DDTC sodium salt aqueous solution as the reaction ionic solution, the molar ratio of Cu(II) to indocyanine green in the nanoparticles is 5:1. The preparation process is as follows:
[0059] (1) Dissolve 0.85 mg of indocyanine green in 6 mL of water, then add 1 mL of DDTC sodium aqueous solution with a concentration of 2.5 mg / mL to obtain mixed solution A, and stir in an oil bath at 50°C for 5 minutes;
[0060] (2) While stirring, 1 mL of a 0.94 mg / mL CuCl2·2H2O salt solution was added dropwise to the obtained mixed solution A over a period of 60 s. The mixing method was magnetic stirring at 1200 rpm and the mixing temperature was 50 °C. After the addition was complete, ultrafiltration purification was performed to obtain an indocyanine green-stabilized CuET nanoparticle dispersion, named Sample 2.
[0061] Example 3
[0062] This embodiment provides indocyanine green-stabilized CuET nanoparticles, and the preparation process is as follows:
[0063] Using CuCl2 and DDTC sodium salt aqueous solution as the reaction ionic solution, the molar ratio of Cu(II) to indocyanine green in the nanoparticles is 40:1. The preparation process is as follows:
[0064] (1) Dissolve 0.11 mg of indocyanine green in 6 mL of water, then add 1 mL of DDTC sodium aqueous solution with a concentration of 2.5 mg / mL, and in an oil bath at 50 °C to obtain mixed solution A;
[0065] (2) While stirring, 1 mL of a 0.94 mg / mL CuCl2·2H2O aqueous solution was added dropwise to the obtained mixed solution A over a period of 60 s. The mixing method was magnetic stirring at 1200 rpm and the mixing temperature was 50 °C. After the addition was complete, ultrafiltration purification was performed to obtain an indocyanine green-stabilized CuET nanoparticle dispersion, named Sample 3.
[0066] Example 4
[0067] This embodiment provides indocyanine green-stabilized CuET nanoparticles, and the preparation process is as follows:
[0068] Using CuCl2 and DDTC sodium salt aqueous solution as the reaction ionic solution, the molar ratio of Cu(II) to indocyanine green in the nanoparticles is 22.5:1. The preparation process is as follows:
[0069] (1) Dissolve 0.2 mg of indocyanine green in 6 mL of water, then add 1 mL of DDTC sodium aqueous solution with a concentration of 2.5 mg / mL, and in an oil bath at 30 °C to obtain mixed solution A;
[0070] (2) While stirring, 1 mL of a 0.94 mg / mL CuCl2·2H2O salt solution was added dropwise to the obtained mixed solution A. The addition time was 60 s, and the mixing method was magnetic stirring at 1200 rpm and the mixing temperature was 30 ℃. After the addition was completed, sample 4 was obtained.
[0071] Experimental Example 1
[0072] Morphological determination of indocyanine green-stabilized CuET nanoparticles: The particle size and distribution of samples 1-4 prepared in Examples 1-4 were measured using a dynamic light scattering instrument. Sample 1 prepared in Example 1 was dropped onto a copper mesh carbon film, dried, and its morphology was observed on a TEM. Figure 2 The results showed that the particle size distribution of samples 1 to 4 prepared in Examples 1 to 4 was relatively uniform, with a distribution range of 40 to 300 nanometers. Figure 3 TEM results for sample 1 showed that the CuET nanocrystals stabilized by indocyanine green had a consistent morphology and uniform particle size.
[0073] Experiment Example 2
[0074] Spectroscopic characterization of nanoparticles: The dispersion of sample 1 prepared in Example 1 was freeze-dried at -50°C for 48 h to obtain a freeze-dried solid powder of indocyanine green-stabilized CuET nanoparticles. XRD and mass spectrometry analyses were performed on this solid sample to confirm its structure and main components. Figure 4The XRD results showed that the peaks at (011), (10-2), (11-2), (11-1), (210), (211), and (220) in the freeze-dried powder of sample 1 were consistent with the results of CuET and the standard card, indicating that CuET exists in the form of nanocrystals in indocyanine green-stabilized CuET nanoparticles. Figure 5 The mass spectrometry results showed that the main components of sample 1 were CuET and indocyanine green.
[0075] Experimental Example 3
[0076] Serum stability test: The sample 1 dispersion prepared in Example 1 was injected into a culture medium containing 10% fetal bovine serum, and the changes in the diameter and PDI of the sample were detected by laser particle size analyzer over 7 days. Figure 6 The results showed that the dispersion of sample 1 had good stability in serum and was suitable for intravenous administration.
[0077] Experiment Example 4
[0078] Mitochondrial membrane potential detection assay: 4T1 cells were seeded into 6-well plates at a density of 400,000 cells / well, with a culture medium volume of 2 mL, and incubated overnight in a hypoxic incubator (37℃, 1% O2). Then, 2 mL of culture medium containing different concentrations of drugs was added, and incubation was continued for 24 h. The cells were stained with JC-1 fluorescent dye, and the mitochondrial membrane potential was assessed by flow cytometry. Figure 7 The results showed that Sample 1 prepared in Example 1 could significantly reduce the mitochondrial membrane potential of 4T1 cells and disrupt the mitochondrial function of tumor cells.
[0079] Experimental Example 5
[0080] Cellular oxygen consumption assay: 4T1 cells were seeded into 6-well plates at a density of 400,000 cells / well, with a culture medium volume of 2 mL. The plates were incubated overnight in a hypoxic incubator (37°C, 1% O2). After cell collection and counting, 400,000 cells were resuspended in 1 mL of PBS. Oxygen consumption was measured using an oxygen-measuring microelectrode. Figure 8 This indicates that Sample 1 prepared in Example 1 can significantly reduce the oxygen consumption of 4T1 tumor cells.
[0081] Experimental Example 6
[0082] Cellular hypoxia detection assay: 4T1 cells were seeded into 6-well plates at a density of 400,000 cells / well, with a culture medium volume of 2 mL, and incubated overnight in a hypoxia incubator (37℃, 1% O2). After staining with a hypoxia probe, cells were collected, and the hypoxia status of the cells was detected by flow cytometry. Figure 9This indicates that Sample 1 prepared in Example 1 can significantly improve the hypoxia of 4T1 tumor cells.
[0083] Experimental Example 7
[0084] Singlet oxygen detection experiment. An aqueous solution containing 10 μg / mL indocyanine green and 5 μM SOSG was prepared by mixing the indocyanine green sample dispersion with the singlet oxygen probe SOSG. The solution was then analyzed using a 1.5 W / cm² flow rate. 2 The SOSG was irradiated with an 808 nm laser for 5 minutes. The fluorescence spectrum of the SOSG was detected every 0.5 minutes using a time-resolved fluorescence spectrometer. Figure 10 This indicates that Sample 1 prepared in Example 1 can effectively generate singlet oxygen and has a photodynamic effect.
[0085] Experimental Example 8
[0086] Female BALB / c mice were subcutaneously inoculated with 4×10 5 A mouse model of subcutaneous breast cancer bearing 4T1 cells was established using mouse 4T1 breast cancer cells. The subcutaneous tumor volume was 50–100 mm. 3 Mice were randomly divided into 5 groups of 8 mice each. Each group received a tail vein injection of saline, indocyanine green solution, or sample 1 dispersion, respectively. The dosage of CuET was 9 mg / kg, and the dosage of indocyanine green was 1.2 mg / kg. The first day of administration was designated as day 0. Drugs were administered every 2 days. On days 0, 2, and 4, 2 hours after administration, the indocyanine green plus light group and the sample 1 plus light group received a 1.5 W / cm² light source. 2 The tumor was irradiated with an 808 nm laser for 10 minutes. Starting from day 0, the long side 'a' and short side 'b' of the subcutaneous tumor in mice were measured every other day using calipers. The tumor volume was calculated using the formula: V = a × b × b / 2. A tumor volume-time curve was plotted. Figure 11 The results showed that Sample 1 prepared in Example 1 could inhibit tumor growth to a certain extent, and its combination with photodynamic therapy could achieve better therapeutic effects.
[0087] Comparative Example 2
[0088] Using CuCl2 and DDTC sodium salt aqueous solution as the reaction ionic solution, the molar ratio of Cu(II) to indocyanine green in the nanoparticles is 5:1. The preparation process is as follows:
[0089] (1) Dissolve 0.85 mg of indocyanine green in 6 mL of water, then add 1 mL of DDTC sodium aqueous solution with a concentration of 2.5 mg / mL, and in an oil bath at 30 °C to obtain mixed solution A;
[0090] (2) While stirring, 1 mL of a 0.94 mg / mL CuCl2·2H2O aqueous solution was added dropwise to the obtained mixed solution A over a period of 60 s. The mixing method was magnetic stirring at 1200 rpm and the mixing temperature was 30 °C. After the addition was complete, the sample of Comparative Example 2 was obtained. The experiment showed that the final solution was still clear, and nanoparticles could not be obtained.
[0091] Comparative Example 3
[0092] Using CuCl2 and DDTC sodium salt aqueous solution as the reaction ionic solution, the molar ratio of Cu(II) to indocyanine green in the nanoparticles is 40:1. The preparation process is as follows:
[0093] (1) Dissolve 0.11 mg of indocyanine green in 6 mL of water, then add 1 mL of DDTC sodium aqueous solution with a concentration of 2.5 mg / mL, and in an oil bath at 70 °C to obtain mixed solution A;
[0094] (2) While stirring, 1 mL of a 0.94 mg / mL CuCl₂·2H₂O aqueous solution was added dropwise to the obtained mixed solution A over a period of 60 s. The mixing method was magnetic stirring at 1200 rpm at a mixing temperature of 70 °C. After the addition was complete, the sample of Comparative Example 3 was obtained. Its particle size distribution results are shown in […]. Figure 2 The resulting nanoparticles had excessively large sizes.
[0095] Comparative Example 4
[0096] Using CuCl2 and DDTC sodium salt aqueous solution as the reaction ionic solution, the molar ratio of Cu(II) to indocyanine green in the nanoparticles is 40:1. The preparation process is as follows:
[0097] (1) Dissolve 0.11 mg of indocyanine green in 6 mL of water, then add 1 mL of DDTC sodium aqueous solution with a concentration of 2.5 mg / mL, and in an oil bath at 30 °C to obtain mixed solution A;
[0098] (2) 1 mL of a 0.94 mg / mL CuCl₂·2H₂O aqueous solution was added dropwise to the obtained mixed solution A while stirring. The addition time was 60 s, and the mixing method was magnetic stirring at 1200 rpm at a mixing temperature of 30 °C. After the addition was complete, the sample of Comparative Example 4 was obtained. Testing revealed that micron-sized crystals were obtained, but stable and uniform nanoparticles could not be formed. The results are shown in […]. Figure 2 .
[0099] Other conditions were the same as in the previous example, except that the molar ratio of Cu(II) to indocyanine green, temperature, and Cu(II) ion concentration were changed. The particle size and distribution of the samples were analyzed, and orthogonal experiments were conducted to obtain the following results: Figure 12The results show that, compared with the concentration of Cu(II) ions, temperature and the molar ratio of Cu(II) to indocyanine green have a greater influence on the formation of CuET nanoparticle dispersions stabilized by indocyanine green. The molar ratio of Cu(II) to indocyanine green is 5-40:1, preferably 5-30:1, and more preferably 5-22.5:1; the preferred reaction temperature range is 40-70℃, and more preferably 50-60℃.
[0100] Comparative Example 5
[0101] The other conditions are the same as in Example 1, except that indocyanine green in Example 1 is replaced with IR780. Following the same method, a precipitate is obtained instead of a nanoparticle dispersion.
[0102] Comparative Example 6
[0103] The other conditions are the same as in Example 1, except that indocyanine green in Example 1 is replaced with tea polyphenols. Following the same method, a precipitate is obtained instead of a nanoparticle dispersion.
[0104] Those skilled in the art will readily understand that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.
Claims
1. An indocyanine green-stabilized CuET nanoparticle, characterized in that, These are nanoparticles formed by stabilizing CuET nanocrystals with indocyanine green, wherein indocyanine green binds to and stabilizes CuET nanocrystals through hydrophobic interactions, forming the indocyanine green-stabilized CuET nanoparticles. The indocyanine green binds and stabilizes CuET nanocrystals through hydrophobic interactions, and is dispersed in water to form an indocyanine green-stabilized CuET nanoparticle dispersion. The preparation method of the indocyanine green-stabilized CuET nanoparticle dispersion includes the following steps: adding a Cu(II) ion salt solution dropwise to a mixed aqueous solution of DDTC salt solution and indocyanine green, stirring simultaneously to allow the reaction to occur, thereby obtaining an indocyanine green-stabilized CuET nanoparticle dispersion; the molar ratio of copper(II) ions to indocyanine green in the Cu(II) ion salt solution is 5~40:1; the reaction temperature is 40~70℃.
2. The nanoparticles as described in claim 1, characterized in that, The particle size of the indocyanine green stabilized CuET nanoparticles is 10~400 nm.
3. An indocyanine green-stabilized CuET nanoparticle dispersion, characterized in that, It comprises indocyanine green stabilized CuET nanoparticles as described in any one of claims 1 to 2, wherein the indocyanine green binds to and stabilizes CuET nanocrystals through hydrophobic interactions, and is dispersed in water to form the dispersion.
4. The nanoparticle dispersion as described in claim 3, characterized in that, The concentration of the indocyanine green-stabilized CuET nanoparticles in the dispersion is 0.1~100 mg / mL.
5. The method for preparing the indocyanine green-stabilized CuET nanoparticle dispersion as described in claim 3 or 4, characterized in that, The process includes the following steps: Cu(II) ion salt solution is added dropwise to a mixed aqueous solution of DDTC salt solution and indocyanine green, and stirring is performed simultaneously to allow the reaction to occur, resulting in an indocyanine green-stabilized CuET nanoparticle dispersion.
6. The preparation method according to claim 5, characterized in that, The molar ratio of DDTC ions to Cu(II) ions is 1.5~2.2:1; the concentration of Cu(II) ions in the dispersion is 0.35~2.78mM.
7. The use of the indocyanine green stabilized CuET nanoparticles as described in claim 1 or 2 and / or the indocyanine green stabilized CuET nanoparticle dispersion as described in claim 3 or 4 in the preparation of a medicament for treating breast cancer.
8. An antitumor drug, characterized in that, Includes indocyanine green stabilized CuET nanoparticles as described in claim 1 or 2 and / or indocyanine green stabilized CuET nanoparticle dispersions as described in claim 3 or 4.