NIR-II nano-particles, preparation method and application of NIR-II nano-particles in tumor treatment by aging induction-photothermal-aging clearance combined therapy

Abstract

The invention provides NIR-II nano-particles, a preparation method and application of the NIR-II nano-particles in treatment of tumors by an aging induction-photothermal-aging clearance combined therapy. The nanoparticle induces tumor cell senescence and down-regulates HSP70 through palbociclib, a photosensitizer IR1061 and quercetin are carried together, mild and efficient tumor ablation is achieved through NIR-II photo-thermal (smaller than or equal to 44 DEG C), senescent cells are removed through quercetin, a heat tolerance barrier is broken through, and relapse is prevented. The scheme has the advantages of deep penetration (3-5cm), high efficiency and low toxicity (the tumor ablation rate is 90%, and the thermal damage is reduced by 60%), multi-mechanism synergy (senescence induction-photo-thermal sensitization-senescence removal) and the like, the system toxicity is remarkably reduced, the life quality of a patient is improved, and a new precise treatment path is provided for solid tumors such as triple-negative breast cancer and the like.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a NIR-II nanoparticle and its application in the treatment of tumors by a senescence induction-photothermal-senescence clearance combined therapy; the present invention also provides a method for preparing the NIR-II nanoparticle. Background Art

[0002] Triple-negative breast cancer (TNBC) lacks estrogen receptors, progesterone receptors, and HER2 expression, resulting in highly invasive, metastatic, and drug-resistant tumors, posing significant challenges to clinical treatment. Traditional treatments such as chemotherapy, radiotherapy, and immunotherapy have limited efficacy and are associated with severe side effects. Photothermal therapy (PTT), which uses photothermal agents to convert light energy into heat to kill tumor cells, has garnered attention due to its non-invasive and precisely controllable properties.

[0003] In recent years, photothermal materials in the near-infrared region II (NIR-II, 1000-1700nm) have become a new hotspot for PTT due to their higher tissue penetration depth and lower tissue scattering. However, tumor cells can develop thermal tolerance by upregulating heat shock proteins (such as HSP70), which limits the effectiveness of photothermal therapy alone, especially in the treatment of deep lesions, which can easily lead to recurrence.

[0004] Although the existing NIR-II photothermal nanoplatform has improved tissue penetration, it has not effectively solved the problem of thermal tolerance. In addition, the nanocarriers combined with chemotherapy or gene therapy are not targeted enough and do not combine the aging regulatory mechanism. Cell senescence induction and clearance (senolytic therapy) provides a new treatment idea for TNBC. CDK4 / 6 inhibitors (such as palbociclib) can induce tumor cell senescence and inhibit their proliferation, while downregulating HSP70 expression to enhance thermal sensitivity; senolytic drugs (such as quercetin) can selectively eliminate senescent cells and prevent the senescence-associated secretory phenotype (SASP). However, relying solely on senescence induction may lead to recurrence due to residual cells, and systemic combination chemotherapy or senolytic drugs have the risk of off-target toxicity.

[0005] To address the above issues, there is an urgent need to develop a multi-mechanism synergistic, precisely controllable TNBC treatment strategy that can lower the treatment temperature to avoid damage to normal tissues, enhance thermal sensitivity through aging regulation, and achieve local and efficient delivery of senolytic drugs, thereby simultaneously solving thermal tolerance, systemic toxicity and recurrence risks. Summary of the Invention

[0006] In view of this, the purpose of the present invention is to provide a near-infrared region II (NIR-II) nanoparticle and its preparation method, as well as the use of the nanoparticle in the treatment of tumors with a senescence induction-photothermal-senescence clearance combination therapy. Unlike existing monotherapy or combination therapy, the NIR-II nanoparticle is designed based on a three-step synergistic strategy of "senescence induction-photothermal sensitization-senescence clearance": palbociclib is used to induce TNBC cell senescence and reduce HSP70, and the NIR-II nanoplatform is used to co-load the photosensitizer IR1061 and quercetin. While killing tumors with mild photothermal (≤44°C), quercetin inhibits HSP70 and clears senescent cells, breaking through the thermal tolerance barrier and preventing recurrence.

[0007] Specifically, the present invention provides a NIR-II nanoparticle comprising a photosensitizer, a senolytic agent, and DSPE-PEG2000-COOH.

[0008] The CAS number of the DSPE-PEG2000-COOH is 474922-20-8.

[0009] In some specific embodiments of the present invention, the photosensitizer is compound IR1061; and the senolytic agent is quercetin.

[0010] The CAS number of the compound IR1061 is 155614-01-0;

[0011] In some specific embodiments of the present invention, the NIR-II nanoparticles are prepared by a nano-coprecipitation method.

[0012] In some specific embodiments of the present invention, the NIR-II nanoparticles are prepared as follows: compound IR1061, quercetin, and DSPE-PEG2000-COOH are dissolved in organic solvents, mixed in a dark environment, and stirred for 4-12 hours; the organic solvent is then removed by vacuum distillation, and then freeze-dried to obtain a powdered solid, namely the NIR-II nanoparticles;

[0013] The compound IR1061, quercetin and DSPE-PEG 2000 The mass ratio of -COOH is 1:(0.5~2):(1.5~4.5).

[0014] In some embodiments of the present invention, the compounds IR1061, quercetin and DSPE-PEG 2000 The mass ratios of -COOH are 1:1:3 respectively.

[0015] In some embodiments of the present invention, the compounds IR1061, quercetin and DSPE-PEG2000 The mass ratios of -COOH are 1:2:3 respectively.

[0016] In some embodiments of the present invention, the compounds IR1061, quercetin and DSPE-PEG 2000 The mass ratios of -COOH are 2:1:3 respectively.

[0017] In some specific embodiments of the present invention, the organic solvent is acetonitrile.

[0018] The present invention also provides a method for preparing NIR-II nanoparticles, comprising the following steps:

[0019] Compound IR1061, quercetin, and DSPE-PEG2000-COOH were dissolved in organic solvents, respectively. The three liquids were mixed and stirred in a dark environment for 4-12 hours. The organic solvent was then distilled off under reduced pressure, and then freeze-dried to obtain a powdered solid, namely, NIR-II nanoparticles.

[0020] The compound IR1061, quercetin and DSPE-PEG 2000 The mass ratio of -COOH is 1:(0.5~2):(1.5~4.5).

[0021] In some specific embodiments of the present invention, the following preparation steps are included:

[0022] Compound IR1061, quercetin, and DSPE-PEG2000-COOH were dissolved in acetonitrile, sonicated, and then mixed in a dark environment and stirred for 4-12 hours. The organic solvent was then distilled off under reduced pressure and freeze-dried to obtain a powdered solid, which is the NIR-II nanoparticles.

[0023] The compound IR1061, quercetin and DSPE-PEG 2000 The mass ratio of -COOH is 1:(0.5~2):(1.5~4.5).

[0024] In some embodiments of the present invention, the compounds IR1061, quercetin and DSPE-PEG 2000 The mass ratios of -COOH are 1:1:3 respectively.

[0025] In some embodiments of the present invention, the compounds IR1061, quercetin and DSPE-PEG 2000 The mass ratios of -COOH are 1:2:3 respectively.

[0026] In some embodiments of the present invention, the compounds IR1061, quercetin and DSPE-PEG2000 The mass ratios of -COOH are 2:1:3 respectively.

[0027] The present invention also provides a method for preparing a NIR-II nanoparticle solution, comprising the following steps:

[0028] Compound IR1061, quercetin, and DSPE-PEG2000-COOH were dissolved in organic solvents, respectively. The three liquids were mixed and stirred in the dark for 4-12 hours. The organic solvent was then removed by vacuum distillation. Nanoparticles were uniformly attached to the inner wall of the rotary evaporator. The resulting nanoparticle solution was dispersed with a pH 6.5 phosphate buffered saline solution.

[0029] The compound IR1061, quercetin and DSPE-PEG 2000 The mass ratio of -COOH is 1:(0.5~2):(1.5~4.5).

[0030] The present invention also provides a pharmaceutical composition for treating tumors using a combined senescence induction-photothermal-senescence clearance therapy, comprising the NIR-II nanoparticles described in any one of the above items or the NIR-II nanoparticles prepared by any one of the above preparation methods.

[0031] In some specific embodiments of the present invention, palbociclib is also included.

[0032] The CAS number of palbociclib is 571190-30-2.

[0033] In some specific embodiments of the present invention, it further comprises a pharmaceutically acceptable carrier or excipient.

[0034] The present invention also provides the use of any of the above-mentioned NIR-II nanoparticles, or the NIR-II nanoparticles prepared by any of the above-mentioned preparation methods, or the pharmaceutical composition for treating tumors by the senescence induction-photothermal-senescence elimination combination therapy as described in any of the above-mentioned methods in treating tumors by the senescence induction-photothermal-senescence elimination combination therapy.

[0035] The senescence-induction-photothermal-senolytic combination therapy is a tumor treatment method based on a three-step synergistic strategy of "senescence induction-photothermal sensitization-senescence clearance": palbociclib induces TNBC cell senescence and reduces HSP70. NIR-II-responsive nanocarriers loaded with photothermal agents and senolytic drugs produce a mild photothermal effect at ≤44°C under NIR-II laser irradiation, ablating tumor tissue and inhibiting HSF1 phosphorylation. The NIR-II nanoplatform co-loads the photosensitizer IR1061 and quercetin. Simultaneously, quercetin inhibits HSP70 and clears senescent cells, breaking through the thermal tolerance barrier and preventing recurrence. In this three-step closed-loop mechanism, palbociclib pretreatment sensitizes the photothermal effect, nanoparticle-mediated NIR-II photothermal therapy directly kills the primary tumor, and quercetin clears residual senescent cells.

[0036] In some embodiments of the invention, the tumor is triple-negative breast cancer.

[0037] The present invention has the following significant advantages and effects compared to the prior art:

[0038] 1. Breakthrough of the bottleneck of traditional photothermal therapy

[0039] Mild temperature and efficient ablation: Palbociclib (a CDK4 / 6 inhibitor) induces tumor cell senescence and directly downregulates HSP70, combined with quercetin to inhibit HSF1 phosphorylation, doubly blocking the heat shock protection pathway, reducing the near-infrared II (NIR-II) photothermal therapy temperature from the traditional >50°C to ≤44°C, achieving a 90% tumor ablation rate while avoiding high temperature toxicity.

[0040] Deep-penetrating precision treatment: Based on the deep tissue penetration ability (up to 3-5 cm) of NIR-II light (1000-1350 nm), combined with the tumor-targeting enrichment characteristics of nanocarriers (IQNPs), it significantly improves the photothermal coverage of deep tumors (such as triple-negative breast cancer) and reduces accidental damage to normal tissues.

[0041] 2. Significantly reduce systemic toxicity

[0042] Local drug release reduces systemic exposure: Targeted enrichment of nanocarriers (IQNPs) increases deep drug concentration, reduces systemic distribution of drugs, reduces normal cell cycle arrest caused by CDK4 / 6 inhibitors and damage to liver and kidney function caused by high doses of senolytic drugs, and there is no significant difference in liver and kidney function indicators compared with the control group.

[0043] Mild temperature reduces thermal damage: Photothermal therapy at ≤44°C reduces the risk of thermal damage to surrounding normal tissues by 60% (compared to traditional >50°C regimen), avoiding inflammation and fibrosis caused by high temperature.

[0044] 3. Multiple mechanisms synergistically block tumor regeneration

[0045] Three-step closed-loop tumor cell elimination: 1) induction of senescence (palbociclib) → 2) photothermal sensitization killing (IR1061+NIR-II) → 3) senescent cell elimination (quercetin). The combination of the three therapies not only kills the primary tumor, but also blocks the tumor regeneration network by eliminating senescent cells, solving the problem that a single therapy cannot block the tumor regeneration network due to its isolated mechanism.

[0046] 4. Improve patients’ quality of life:

[0047] Reduced side effects: IQNPs-mediated local high-concentration release of NIR-II photothermal therapy and senolytic drugs significantly reduced side effects such as liver and kidney damage during treatment, resulting in no significant difference in the patients' liver and kidney function indicators compared with the control group.

[0048] Personalized treatment plan: Based on the high heterogeneity and drug resistance characteristics of TNBC, a new paradigm of personalized treatment with high efficiency and low toxicity is provided, bringing new hope to patients with TNBC and other solid tumors.

[0049] In summary, the NIR-II nanoparticles and pharmaceutical compositions provided by the present invention, by integrating the penetration advantages of NIR-II, aging-regulated enhanced sensitivity and senolytic targeted clearance, provide a new path for the precise treatment of TNBC and other solid tumors with high efficiency and low toxicity, significantly improving the therapeutic effect, reducing the recurrence rate and side effects, and have important clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0051] Figure 1 Schematic diagram of the IQ preparation process using nano-coprecipitation method.

[0052] Figure 2 The particle sizes of TEM and 2:1IQ NPs.

[0053] Figure 3 The particle sizes of TEM and 1:1IQ NPs.

[0054] Figure 4 The particle size of NPs is 1:2IQ.

[0055] Figure 5Zeta potentials of IQ NPs with Qu, IR, and different IR-Qu ratios.

[0056] Figure 6 Drug loading capacity of IQ NPs with different IR and Qu ratios.

[0057] Figure 7 Encapsulation efficiency of IQ NPs with different IR and Qu ratios.

[0058] Figure 8 Images of IQ NPs (50 μg / ml) in PBS.

[0059] Figure 9 Transmission electron microscopy (TEM) of IQ NPs.

[0060] Figure 10 UV-Vis-NIR spectra of quercetin in acetonitrile and IQ NPs and IR1061 in PBS.

[0061] Figure 11 (a) shows the 14-day stability of IQ NPs (25 μg / m1) in different dispersion media, and (b) shows the particle size of IQ NPs in different dispersion media at 0, 7, and 14 days.

[0062] Figure 12 (a) is the test of the uptake ability of normal and senescent 4T1 cells for IQ NPs within 8 hours; (b) is the quantitative uptake graph of normal and senescent 4T1 cells within 8 hours.

[0063] Figure 13 Flow cytometry was used to quantitatively analyze the uptake capacity of 4T1 cells within 8 hours after senescence.

[0064] Figure 14 (a) is the test of different concentrations of IQ NPs with or without laser irradiation (1060nm, 1.0W / cm 2 ) of 4T1 cell survival rate; (b) to test the effect of different concentrations of IR and IQ NPs with or without laser irradiation (1060nm, 1.0W / cm 2 ) in the survival rate of senescent 4T1 cells (mean ± SD, n = 3, two-way ANOVA test, *p < 0.05; ****p < 0.0001 compared with the control group).

[0065] Figure 15 Representative flow cytometry images show the apoptosis levels of 4T1 cells after various treatments (using Annexin V-FITC / PI apoptosis detection kit).

[0066] Figure 16CLSM images of 4T1 cells stained with Calcein AM / PI after receiving different treatments.

[0067] Figure 17 Schematic diagram of the treatment plan for mice bearing subcutaneous 4T1 tumors.

[0068] Figure 18 Time dependence of photoacoustic imaging (PAI) of tumor sites after intravenous injection of IQNPs in normal and senescent tumor-bearing mice.

[0069] Figure 19 For quantitative analysis of photoacoustic imaging signals.

[0070] Figure 20 (a) shows the changes in tumor volume during treatment (mean ± standard deviation, N = 5, two-way ANOVA test, compared with the control group, ****p < 0.0001); (b) shows the changes in mouse body weight during treatment.

[0071] Figure 21 The 4T1 tumor-bearing mice treated with IR nanoparticles and IQ nanoparticles were irradiated with 1060 nm laser (1.0 w / cm 2 ) thermal imaging changes over time.

[0072] Figure 22 (a) is the image of the hemolysis experiment of IQ nanoparticles with different concentrations; (b) is the quantitative analysis of the hemolysis experiment (mean ± standard deviation, N = 3, one-way ANOVA test, compared with the "100" group, ****p < 0.0001). DETAILED DESCRIPTION

[0073] The following will be combined with the accompanying drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention and should not be used to limit the scope of protection of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0074] Example 1: Preparation of NIR-II nanoparticles (IQ nanoparticles)

[0075] The specific steps are as follows:

[0076] IQ nanoparticles were prepared by nanocoprecipitation method using IR1061, quercetin and DSPE-PEG2000-COOH.

[0077] The specific preparation steps of IQ nanoparticles with IR (IR1061): Qu (quercetin) = 1:2 (mass ratio) are as follows: IR1061 (1 mg, CAS number 155614-01-0), quercetin (2 mg) and DSPE-PEG2000-COOH (3 mg, CAS number 474922-20-8) were dissolved in 2 mL of acetonitrile, and the above liquids were placed in an ultrasonic water bath for 10 minutes. Then, the three liquids were mixed and stirred in a dark environment at room temperature and a stirring rate of 1000 rpm for 4-12 hours. The acetonitrile was removed by rotary distillation, and then a phosphate buffered saline (PBS, pH 6.5) solution was added to obtain IQ nanoparticles, which were stored in a refrigerator at 4°C for the next experiment.

[0078] The IQ nanoparticles with IR (IR1061): Qu (quercetin) = 1:1 (mass ratio) and the IQ nanoparticles with IR (IR1061): Qu (quercetin) = 2:1 (mass ratio) only changed the raw material mass ratio, and the other preparation steps were the same as the above preparation steps.

[0079] Schematic diagram of the preparation steps Figure 1 shown.

[0080] Example 2: Characterization of IQ Nanoparticles

[0081] In order to determine the optimal ratio of IR1061 (IR) to quercetin (Qu) in IQ nanoparticles, three batches of IQ nanoparticles with IR to Qu ratios of 1:2, 1:1, and 2:1 were prepared in Example 1. The particle sizes of the IQ nanoparticles with IR to Qu ratios of 2:1 and 1:1 were approximately 190.1±40 nm ( Figure 2 shown) and 122.4±20nm( Figure 3 As shown), the particle size of IQ nanoparticles with a ratio of 1:2 is approximately 164.2±20 nm ( Figure 4 As shown). The ζ potential of pure Qu is about -25mV, and the ζ potential of IR is about -20mV; for different proportions of IQ nanoparticles, the ζ potentials are -6mV, -20mV and -22mV respectively ( Figure 5 This clearly shows that the IQ nanoparticles with IR to Qu ratios of 1:2 and 2:1 have higher zeta potentials, which means they are more stable. Subsequently, this example measured the drug loading and encapsulation efficiency of IQ nanoparticles with different ratios. The drug loading rates of IR in IQ nanoparticles with different ratios were 20.44%, 23.04% and 37.98%, respectively, while the drug loading rates of Qu were 19.58%, 36.59% and 20.45%, respectively ( Figure 6In terms of encapsulation efficiency, the encapsulation efficiency of IR was 87.12%, 83.86% and 88.98% respectively, and the encapsulation efficiency of Qu was 64.25%, 76.43% and 46.36% respectively ( Figure 7 ). Based on these results, IQ nanoparticles with a ratio of 1:2 showed better drug loading performance for IR compared to IQ nanoparticles with a ratio of 1:1. IQ nanoparticles with a ratio of 1:2 exhibited superior drug loading capacity for Qu compared to IQ nanoparticles with a ratio of 2:1. In addition, a lower dose of IR can effectively mitigate the toxicity caused by high doses of the organic dye IR, thereby ensuring that the nanoparticles have higher biosafety. In addition, the encapsulation efficiency of IR in the nanoparticles was higher in IQ nanoparticles with a ratio of 1:2 than in IQ nanoparticles with the other two ratios. Therefore, it can be determined that IQ nanoparticles with a ratio of 1:2 IR to Qu are the optimal core ratio.

[0082] The prepared nanoparticle solution with a ratio of IR to Qu of 1:2 showed a clear and transparent yellow-green liquid ( Figure 8 IQ nanoparticles were uniformly dispersed in PBS to form spherical particles with a diameter of approximately 164.2 ± 30 nm ( Figure 9 ).

[0083] UV-Vis-NIR spectra were tested for IR, Qu and IQ nanoparticles (the ratio of IR to Qu was 1:2, and the IQ nanoparticles shown below were all nanoparticles with this ratio). Figure 10 ) shows that the maximum absorption peak of Qu is at 382 nm and the maximum absorption peak of IR is at 1073 nm, which confirms that IR and Qu have been successfully co-loaded into IQ nanoparticles. Subsequently, IQ nanoparticles were diluted in water, PBS, fetal bovine serum (FBS) and DMEM culture medium to a concentration of 25 μg / ml. The particle size of IQ nanoparticles increased from 190.1±30 nm on day 0 to 220±30 nm on day 14 ( Figure 11 ), and there was no obvious precipitation phenomenon, which indicated that the nanoparticles had excellent stability.

[0084] Example 3: In vitro killing effect test of IQ nanoparticles

[0085] The following examples demonstrate the uptake and killing effects of IQNPs and their respective components in cells.

[0086] The cells were divided into a normal group (N) and a senescent group (S) according to whether they were induced to age for 3 days in advance with 2.5 μM concentration of palbociclib. The IQ NPs were tested to see whether they could be successfully taken up by 4T1 cells within a certain period of time to determine the optimal treatment time. First, confocal laser scanning microscopy (CLSM) was used to observe that a small amount of red fluorescence (cy5.5) appeared around the cells in both groups 2-4 hours after administration. The uptake of IQNPs by the S group was slightly higher than that of the N group. This may be due to the insignificant drug release rate within 2-4 hours and the increased membrane permeability of senescent cells, which led to an increase in the uptake of IQNPs. The uptake of the N group reached its maximum within 4-8 hours ( Figure 12 ). Subsequently, the uptake capacity of senescent cells was quantitatively analyzed by flow cytometry ( Figure 13 ), the results were similar to those observed under CLSM. Therefore, in the subsequent cell experiments, 8 hours was selected as the administration time. Subsequently, the cytotoxicity of IQNPs under aging conditions was evaluated using CCK-8 assay ( Figure 14 When the cells were not senescent and not irradiated with laser light, the survival rates of both groups exceeded 80%, even at a high concentration of 25 μg / mL. In contrast, under senescent conditions, the cell-killing effect of IQ NPs was more pronounced. At a drug concentration of 25 μg / mL, the cell survival rate was less than 35%. Therefore, 25 μg / mL was set as the concentration of IQ NPs administered to cells.

[0087] To further verify the above results, cell death and apoptosis assays were also performed under mild photothermal therapy. Flow cytometry was used to detect the combined effects of S and Qu, as well as the ability of IQNPs±Laser to induce apoptosis ( Figure 15 ). S and Qu showed low toxicity, with apoptosis rates of 4.52% and 9.81%, respectively. The apoptosis rate in the S+Qu group increased to 11.64%. Compared with the N group, S+IR induced apoptosis in some cells through the photothermal effect after laser irradiation. After loading Qu, the apoptosis rate in the S+IQ group reached 33.7% after laser irradiation. As an HSP inhibitor, Qu can effectively eliminate the heat resistance effect of tumor cells and improve the photothermal killing effect. After cell senescence, the use of Qu to eliminate senescent cells led to an increase in the apoptosis rate in the later stage. Combined with the inhibitory effect of Qu on HSP, the apoptosis rate even reached 33.7%. Finally, cell viability and death were determined ( Figure 16) It was observed that cells in each group survived remarkably well without laser irradiation; however, a small amount of red fluorescence appeared in the S+IR±Laser group, indicating that mild PTT following cellular senescence was ineffective in producing significant cell death. A large number of cells in the S+IQ group had already died. Based on these results, IQNPs utilize the photothermal effect of IR1061, combined with Qu to eliminate senescent cells and weaken the heat resistance of tumor cells, thereby exerting a stronger apoptotic effect at the cellular level, validating the excellent tumor-killing ability of S+IQNPs.

[0088] Example 4: In vivo killing effect test of IQ nanoparticles

[0089] In this study, 1 million 4T1 cells were injected subcutaneously into the right hind leg of nude mice to form tumors. Subsequently, 100 mg / kg of palbociclib was gavaged into the nude mice for 7 days to establish a mouse model carrying aging tumors. In order to further verify the tumor targeting and oncolytic effect of IQ nanoparticles (IQNPs), this example injected IQ nanoparticles into mice through the tail vein and monitored the enrichment of IQ nanoparticle signals at the tumor site, as well as the changes in tumor size and weight of the mice. Finally, the mice were euthanized (the treatment plan flow chart is shown in the figure). Figure 17 First, this example verifies the targeting and imaging capabilities of 1Q nanoparticles in mice in groups N and S. Photoacoustic imaging results and quantitative analysis data of triple-negative breast cancer model mice after tail vein injection of IQ nanoparticles (containing 10 μg IR1061) were obtained ( Figure 18 and 19As shown). Photoacoustic signals showed that in the early stage of tail vein injection, the S group mice had a better uptake of IQ nanoparticles. This may be because the defense ability of aging tumor cells decreases, making it easier for IQ nanoparticles to penetrate. At the same time, in the late stage of tail vein injection, the enrichment of the S group and the N group was similar, and both groups reached the maximum uptake of IQ nanoparticles. Therefore, IQ nanoparticles can be well enriched in the tumor site within 20 hours and have good photoacoustic imaging effects. Secondly, after inducing tumor senescence, phosphate buffered saline (PBS, for the N group), quercetin, IR nanoparticles (containing 10μg IR1061) and IQ nanoparticles (containing 10μg IR1061) were injected through the tail vein, and then the tumor was irradiated with a 1060 nm laser (1.0w / cm2, 10min). According to the 14-day treatment plan, the tumor volume of the N group reached 958.48mm3, while the tumor volume of the S+Qu group was 689.95mm3. Obviously, the tumor volume of the S group and the S+Qu group was lower than that of the N group. In addition, the tumor growth slopes of the other two groups containing quercetin were relatively gentle compared to the other groups, indicating that the combination of tumor senescence and quercetin has a certain inhibitory effect on tumor growth. Subsequently, when the IQ nanoparticles were irradiated with laser, the effect of mild photothermal therapy was enhanced due to the addition of quercetin, which was able to specifically inhibit and eradicate tumor growth. It is worth noting that the final tumor volume of the S+IQ+laser group was only 29.376mm 3 , the tumor was effectively eliminated ( Figure 20 This demonstrates that by inducing tumor senescence, reducing tumor heat resistance, and supplementing with aging agents and heat shock protein inhibitors to enhance mild photothermal therapy, excellent anti-cancer effects can be achieved without causing significant harm to mice ( Figure 21 By monitoring the changes in tumor temperature, the efficiency of mild photothermal therapy after laser irradiation was studied. After laser irradiation, the body temperature of the tumor site of the mouse was within the temperature range of mild photothermal therapy, which shows that IQ nanoparticles have the effect of mild photothermal therapy in vivo ( Figure 22 ). Finally, the biosafety of IQ nanoparticles related to hemolysis in vivo was verified. As the concentration of IQ nanoparticles increased, the hemolysis rate increased from 1.65% to 3.62%. Considering that the nanoparticles may dissolve during the experiment, the actual hemolysis rate should be lower. The above experiments demonstrated that IQ nanoparticles have good targeted enrichment ability and photoacoustic imaging capabilities. In the in vivo treatment experiment, the S+IQ+laser group showed the best anti-tumor effect and stable biochemical safety.

[0090] The above description is merely a preferred embodiment of the present application and an illustration of the technical principles employed. Those skilled in the art should understand that the scope of the invention herein is not limited to the technical solutions formed by the specific combination of the above-mentioned technical features, but also encompasses other technical solutions formed by any combination of the above-mentioned technical features or their equivalents without departing from the inventive concept. For example, the above-mentioned features may have similar functions to (but not limited to) those disclosed in this application.

Claims

1. A NIR-II nanoparticle, characterized in that: Contains a photosensitizer, a senolytic agent, and DSPE-PEG2000-COOH.

2. The NIR-II nanoparticles according to claim 1, wherein The photosensitizer is compound IR1061; the aging clearance agent is quercetin.

3. The NIR-II nanoparticle according to any one of claims 1 to 2, characterized in that The NIR-II nanoparticles are prepared by a nano-coprecipitation method.

4. The NIR-II nanoparticles according to claim 3, wherein The NIR-II nanoparticles are prepared by dissolving the compound IR1061, quercetin, and DSPE-PEG2000-COOH in organic solvents, mixing the three liquids in a dark environment and stirring for 4-12 hours; then removing the organic solvent by vacuum distillation, followed by freeze-drying to obtain a powdered solid, namely the NIR-II nanoparticles; The compound IR1061, quercetin and DSPE-PEG 2000 The mass ratio of -COOH is 1:(0.5~2):(1.5~4.5).

5. A method for preparing NIR-II nanoparticles, characterized in that: The method comprises the following preparation steps: Compound IR1061, quercetin, and DSPE-PEG2000-COOH were dissolved in organic solvents, mixed in a dark environment, and stirred for 4-12 hours. The organic solvent was then removed by vacuum distillation, followed by freeze-drying to obtain a powdered solid, namely, NIR-II nanoparticles. The compound IR1061, quercetin and DSPE-PEG 2000 The mass ratio of -COOH is 1:(0.5~2):(1.5~4.5).

6. The method for preparing NIR-II nanoparticles according to claim 5, wherein: The method comprises the following preparation steps: Compound IR1061, quercetin, and DSPE-PEG2000-COOH were dissolved in acetonitrile, sonicated, and then mixed in the dark and stirred for 4-12 hours. The organic solvent was then distilled off under reduced pressure, and the mixture was lyophilized to obtain a powdered solid, namely, NIR-II nanoparticles. The compound IR1061, quercetin and DSPE-PEG 2000 The mass ratio of -COOH is 1:(0.5~2):(1.5~4.5).

7. A pharmaceutical composition for treating tumors by a combined senescence induction-photothermal-senescence elimination therapy, characterized in that: The invention relates to a photothermal agent according to any one of claims 1 to 4 or NIR-II nanoparticles prepared by the preparation method according to any one of claims 5 to 6.

8. The pharmaceutical composition for treating tumors by senescence induction-photothermal-senescence elimination combined therapy according to claim 7, characterized in that: Also contains palbociclib.

9. Use of the NIR-II nanoparticles according to any one of claims 1 to 4, or the NIR-II nanoparticles prepared by the preparation method according to any one of claims 5 to 6, or the pharmaceutical composition for treating tumors with a senescence-induction-photothermal-senescence elimination combination therapy according to any one of claims 7 to 8 in treating tumors with a senescence-induction-photothermal-senescence elimination combination therapy.

10. The use according to claim 9, characterized in that The tumor is triple-negative breast cancer.

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