Optical-magnetic visualized bio-hybrid nanosystem, preparation method and application thereof

Abstract

The application discloses a kind of optical magnetic visualization biological hybrid nanosystem and its preparation method and application, the optical magnetic visualization biological hybrid nanosystem includes platelet membrane and the tannic acid manganese complex of losartan loading wrapped in the platelet membrane;The optical magnetic visualization biological hybrid nanosystem of the application can promote the deep penetration of drug and immune cell, and realizes tumor targeted therapy using the biological tropism of platelet membrane, in addition, it can also provide visual tracking for precision targeted therapy by fluorescence imaging and magnetic imaging dual imaging mode.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a photomagnetic visualization bio-hybrid nanosystem, its preparation method, and its applications. Background Technology

[0002] Overcoming the interference of the tumor microenvironment and efficiently delivering drugs to penetrate deep into the tumor is an effective way to restore tumor drug sensitivity and induce stronger and more specific anti-tumor immune responses. Tumor-associated fibroblasts (CAFs), abundant in tumor tissue, provide a favorable environment for tumor growth. CAFs play a crucial role in cancer. CAFs can produce various extracellular matrix proteins and regulatory molecules, shaping a dense tumor extracellular matrix that forms a penetration barrier for drugs and therapeutic immune cells, hindering their penetration into deeper tumor tissues and thus reducing the effectiveness of tumor treatment. CAFs can prevent the infiltration and migration of immune cells by reshaping the contact barrier between immune cells and cancer cells. CAF reprogramming to regulate the microenvironment and assist other treatment methods is a novel strategy for overcoming tumor drug resistance and enhancing immunity.

[0003] CAF modulation holds great potential in photothermal therapy-induced anti-tumor immunity and the generation of long-term immune memory. The aim of CAF modulation is to enhance the accumulation and penetration of photosensitizers in tumors, thereby strengthening photothermal therapy-induced T-cell-mediated anti-tumor immunity, reshaping the tumor immunosuppressive microenvironment, and effectively inhibiting tumor recurrence and metastasis. However, traditional delivery strategies still suffer from insufficient distribution specificity and poor solubility, limiting the development of this novel tumor treatment strategy. Therefore, how to simultaneously modulate CAF through precise targeted delivery systems while enhancing the accumulation and penetration of photosensitizers in solid tumors, achieving CAF-modulated photothermal-immunotherapy, is a major challenge currently faced.

[0004] In view of this, the present invention is hereby proposed. Summary of the Invention

[0005] The purpose of this invention is to provide a photomagnetic visualization biohybrid nanosystem, its preparation method, and its application. This photomagnetic visualization biohybrid nanosystem can promote the deep penetration of drugs and immune cells, and utilize the biotaxis of platelet membranes to achieve tumor targeted therapy. In addition, it can provide visual tracking for precise targeted therapy through dual imaging modes of fluorescence imaging and magnetic imaging.

[0006] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted:

[0007] The first aspect of the present invention provides a photomagnetic visualization biohybrid nanosystem, the photomagnetic visualization biohybrid nanosystem comprising a platelet membrane and a manganese tannate (Mn-TA) complex loaded with losartan encapsulated within the platelet membrane.

[0008] Preferably, the platelet membrane is further loaded with neoindocyanine green.

[0009] A second aspect of this invention provides a method for preparing the above-mentioned photomagnetic visualization bio-hybrid nanosystem, the method comprising the following steps:

[0010] (a) Under ultrasonic conditions, losartan solution was added dropwise to polyvinylpyrrolidone solution and ultrasonic treatment was continued for a period of time to obtain losartan nanoparticle solution.

[0011] (b) Tannic acid solution and manganese chloride solution were added dropwise to the losartan nanoparticle solution and ultrasonic treatment was performed. The pH value was then adjusted to alkaline and ultrasonic treatment was continued. After centrifugation, washing and drying, the losartan-loaded Mn-TA complex was obtained.

[0012] (c) Add neoindocyanine green to the platelet membrane solution, incubate and centrifuge to obtain a platelet membrane loaded with neoindocyanine green;

[0013] (d) The platelet membrane loaded with new indocyanine green was encapsulated with a losartan-loaded Mn-TA complex by sonication to obtain the photomagnetic visualization biohybrid nanosystem.

[0014] Preferably, in step (a), the volume ratio of losartan solution to polyvinylpyrrolidone solution is 1:(15-25);

[0015] The concentration of losartan solution is 40–60 mg / ml; the concentration of polyvinylpyrrolidone solution is 0.8%–1.2%. Preferably, in step (b), the volume ratio of losartan nanoparticle solution, tannic acid solution, and manganese chloride solution is (7–9):(2–4):(0.8–1.2).

[0016] The concentration of tannic acid solution is 8–12 mg / ml; the concentration of manganese chloride solution is 4–6 mg / ml.

[0017] Preferably, in step (b), the pH value is adjusted to 7.5–9.

[0018] Preferably, in step (c), the mass ratio of the protein content in the platelet membrane to that indocyanine green is (2-3):1.

[0019] Preferably, in step (c), the incubation temperature is 0–8°C and the incubation time is 10–18 h.

[0020] Preferably, in step (d), the mass ratio of platelet membrane protein content to losartan is 1:(4-6).

[0021] The third aspect of this invention provides an application of the above-described photomagnetic visualization biohybrid nanosystem or the photomagnetic visualization biohybrid nanosystem prepared by the above-described preparation method in the preparation of tumor treatment products.

[0022] Compared with the prior art, the beneficial effects of the present invention include at least the following:

[0023] This invention's photomagnetic visualization biohybrid nanosystem promotes deep penetration of drugs and immune cells, utilizes the biotaxis of platelet membranes for tumor-targeted therapy, and provides visual tracking for precise targeted therapy through dual imaging modes of fluorescence and magnetic imaging. Specifically, losartan nanoparticles, acting as tumor microenvironment modulators, are encapsulated in the platelet membrane delivery system, increasing drug loading, regulating and reprogramming the cancer cell environment (CAF), reducing the tumor extracellular matrix, promoting deep penetration of drugs and immune cells, and enhancing tumor treatment efficacy. The photothermal reagent, indocyanine green IR820, is inherently fluorescent, and the nanoparticles utilize manganese ion magnetic imaging, providing visual tracking for precise targeted therapy through dual imaging modes of fluorescence and magnetic imaging. This invention's photomagnetic visualization biohybrid nanosystem not only integrates the advantages of delivery systems combining cell membranes and nanomaterials, but also provides a functional balance between loading and carrier activity through the combination of cell-inspired carriers and nanomaterial systems, while simultaneously utilizing the biotaxis of platelet membranes for tumor-targeted therapy.

[0024] The photomagnetic visualization biohybrid nanosystem of this invention can achieve CAF-regulated synergistic photothermal-immunotherapy, making up for the shortcomings of single therapy in some aspects of anti-tumor treatment; in addition, the photomagnetic visualization biohybrid nanosystem of this invention is simple to implement, highly efficient, and effective, meeting the requirements of application. Attached Figure Description

[0025] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the accompanying drawings used in the description of the specific embodiments or the prior art will be briefly introduced below.

[0026] Figure 1 This is a particle size distribution diagram of different materials in the experimental examples of this invention;

[0027] Figure 2 These are zeta potential analysis diagrams of different materials in the experimental examples of this invention;

[0028] Figure 3 These are in vitro thermal imaging images of different nanoparticles under laser irradiation in the experimental examples of this invention;

[0029] Figure 4These are the in vitro photothermal heating curves of different nanoparticles under laser irradiation in the experimental examples of this invention;

[0030] Figure 5 This describes the temperature changes of IR-PV@LNP under laser irradiation for three irradiation cycles in the experimental examples of this invention.

[0031] Figure 6 These are the flow cytometry analysis results from the experimental examples of this invention;

[0032] Figure 7 This describes the T cell proliferation in the experimental examples of this invention;

[0033] Figure 8 These are the results of laser confocal microscopy analysis in the experimental examples of this invention;

[0034] Figure 9 This describes the penetration of IR and IR-PV into tumor spheres in experimental examples of this invention;

[0035] Figure 10 This is an image of fluorescence distribution in mice after tail vein injection of nanoparticles in an experimental example of this invention.

[0036] Figure 11 This is a magnetic resonance image of a tumor-bearing mouse 24 hours after intravenous injection of IR-PV@LNP in an experimental example of this invention;

[0037] Figure 12 These are tumor growth curves for different treatment groups in the experimental examples of this invention;

[0038] Figure 13 These are in vitro images of tumors from different treatment groups in the experimental examples of this invention. Detailed Implementation

[0039] The embodiments of the technical solution of the present invention will be described in detail below with reference to the examples. The following embodiments are only used to illustrate the technical solution of the present invention more clearly, and are therefore only examples, and should not be used to limit the scope of protection of the present invention.

[0040] It should be noted that, unless otherwise stated, the technical or scientific terms used in this application should have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0041] This invention provides a photomagnetic visualization biohybrid nanosystem, which includes a platelet membrane and a losartan-loaded Mn-TA complex encapsulated within the platelet membrane.

[0042] In one embodiment, the platelet membrane is also loaded with neoindocyanine green.

[0043] Another embodiment of the present invention provides a method for preparing the above-mentioned photomagnetic visualization bio-hybrid nanosystem, the method comprising the following steps:

[0044] (a) Under ultrasonic conditions, losartan solution was added dropwise to polyvinylpyrrolidone solution and ultrasonic treatment was continued for a period of time to obtain losartan nanoparticle solution.

[0045] (b) Tannic acid solution and manganese chloride solution were added dropwise to the losartan nanoparticle solution and ultrasonic treatment was performed. The pH value was then adjusted to alkaline and ultrasonic treatment was continued. After centrifugation, washing and drying, the losartan-loaded Mn-TA complex was obtained.

[0046] (c) Add neoindocyanine green to the platelet membrane solution, incubate and centrifuge to obtain a platelet membrane loaded with neoindocyanine green;

[0047] (d) The platelet membrane loaded with new indocyanine green was encapsulated with a losartan-loaded Mn-TA complex by sonication to obtain the photomagnetic visualization biohybrid nanosystem.

[0048] In one embodiment, in step (a), the volume ratio of losartan solution to polyvinylpyrrolidone solution can be any value in the range of 1:(15 to 25), such as 1:15, 1:20 or 1:25.

[0049] In one embodiment, the concentration of the losartan solution can be any concentration from 40 to 60 mg / ml, for example, 40 mg / ml, 50 mg / ml or 60 mg / ml; the concentration of the polyvinylpyrrolidone solution can be any concentration from 0.8% to 1.2%, for example, 0.8%, 1% or 1.2%.

[0050] In one embodiment, in step (b), the volume ratio of losartan nanoparticle solution, tannic acid solution and manganese chloride solution can be any ratio of (7-9):(2-4):(0.8-1.2), specifically 7:2:1.2, 9:4:0.8 or 8:3:1.

[0051] In one embodiment, the concentration of the tannic acid solution can be any concentration from 8 to 12 mg / ml, specifically 8 mg / mL, 10 mg / ml or 12 mg / ml; the concentration of the manganese chloride solution can be any concentration from 4 to 6 mg / ml, specifically 4 mg / ml, 5 mg / ml or 6 mg / ml.

[0052] In one embodiment, in step (b), the pH value is adjusted to 7.5 to 9, specifically to 7.5, 8, or 9.

[0053] In one embodiment, in step (c), the mass ratio of the protein content in the platelet membrane to that of neoindocyanine green can be any ratio of (2 to 3):1, specifically 2:1 or 3:1.

[0054] In one embodiment, in step (c), the incubation temperature can be 0–8°C and the incubation time can be 10–18 h.

[0055] In one embodiment, in step (d), the mass ratio of the protein content in the platelet membrane to losartan can be any value in the range of 1:(4 to 6), specifically 1:5 or 1:4.

[0056] Another embodiment of the present invention provides an application of the above-described photomagnetic visualization biohybrid nanosystem or the photomagnetic visualization biohybrid nanosystem prepared by the above-described preparation method in the preparation of tumor treatment products.

[0057] The technical solution of the present invention will be further described in detail below through specific embodiments.

[0058] Example 1

[0059] This embodiment describes a method for preparing a photomagnetically visualized bio-hybrid nanosystem, which includes the following steps:

[0060] (a) Under ultrasonic conditions, 400 μl of losartan solution (50 mg / ml) was added dropwise to 8 ml of 1% polyvinylpyrrolidone solution, and ultrasonic treatment was continued at 4 °C for 1 min to obtain losartan nanoparticle (denoted as Los NP) solution.

[0061] (b) Add 3 ml of tannic acid solution (10 mg / ml) and 1 ml of manganese chloride solution (5 mg / ml) to the losartan nanoparticle solution and sonicate at 4 °C for 1 min. Then add sodium bicarbonate solution (50 mg / ml) to adjust the pH to 8.0, sonicate at 4 °C for 1 min, centrifuge at 12000 g for 10 min, wash three times, and freeze dry to obtain the losartan-loaded Mn-TA complex (denoted as LNP).

[0062] (c) Blood was collected from BALB / c mice and centrifuged twice at 200g for 15 min to completely remove red blood cells. The supernatant was then centrifuged at 1000g for 30 min to precipitate platelets. After washing three times with PBS, platelets were collected up to the target number of 1×10⁸ and placed in 50 mL of hypotonic solution (prepared by adding 10 μL of benzyl sulfonyl fluoride to 1 mL of pure water) and lysed at 4°C for 24 hours. After lysis, the lysate was subjected to repeated freeze-thaw cycles in liquid nitrogen and at 37°C until the degree of cell rupture exceeded 70%. The lysate was subjected to gradient centrifugation. First, it was centrifuged at 2000g for 10 min at 4°C, and the supernatant was collected. Then, it was centrifuged at 20000g for 30 min, and the precipitate was collected. This precipitate is the platelet membrane fragment. The collected platelet membrane fragments were placed in an ultrasonic disruptor and subjected to ultrasonic treatment at 4°C with a power of 750W, a frequency of 20KHz, and an amplitude of 30%. Each ultrasonic treatment lasted for 5 minutes and was repeated twice to obtain nanoscale platelet membranes (denoted as PV).

[0063] Add 1 mL of neoindocyanine green (4 mg / mL) to 1 mL of platelet membrane solution (protein content 5 mg / mL) and incubate at 4 °C for 12 h. Centrifuge at 20,000 rpm for 30 min to obtain a platelet membrane loaded with neoindocyanine green (denoted as IR-PV).

[0064] (d) Mix 500 μL of LNP (losartan content: 4 mg / mL) prepared above with 500 μL of IR-PV (protein concentration: 1 mg / mL, IR: 200 μg), sonicate at 4 °C for 5 min, and repeat twice with a 5 min interval to obtain the photomagnetic visualization biohybridization nanosystem (denoted as IR-PV@LNP).

[0065] Comparative Example 1

[0066] This comparative example demonstrates a method for preparing a photomagnetically visualized bio-hybrid nanosystem, which includes the following steps:

[0067] (a) Under ultrasonic conditions, 400 μl of 1% polyvinylpyrrolidone solution was added dropwise to 8 ml of 1% polyvinylpyrrolidone solution, and ultrasonic treatment was continued at 4 °C for 1 min to obtain a mixture.

[0068] (b) Add 3 ml of tannic acid solution (10 mg / ml) and 1 ml of manganese chloride solution (5 mg / ml) to the mixture and sonicate at 4 °C for 1 min. Then add sodium bicarbonate solution (50 mg / ml) to adjust the pH to 8.0, sonicate at 4 °C for 1 min, centrifuge at 12000g for 10 min, wash three times, and freeze dry to obtain the Mn-TA complex (denoted as NP).

[0069] (c) Blood was collected from BALB / c mice and centrifuged twice at 200g for 15 min to completely remove red blood cells. The supernatant was then centrifuged at 1000g for 30 min to precipitate platelets. After washing three times with PBS, platelets were collected up to the target number of 1×10⁸ and placed in 50 mL of hypotonic solution (prepared by adding 10 μL of benzyl sulfonyl fluoride to 1 mL of pure water) and lysed at 4°C for 24 hours. After lysis, the lysate was subjected to repeated freeze-thaw cycles in liquid nitrogen and at 37°C until the degree of cell rupture exceeded 70%. The lysate was subjected to gradient centrifugation. First, it was centrifuged at 2000g for 10 min at 4°C, and the supernatant was collected. Then, it was centrifuged at 20000g for 30 min, and the precipitate was collected. This precipitate is the platelet membrane fragment. The collected platelet membrane fragments were placed in an ultrasonic disruptor and subjected to ultrasonic treatment at 4°C with a power of 750W, a frequency of 20KHz, and an amplitude of 30%. Each ultrasonic treatment lasted for 5 minutes and was repeated twice to obtain nanoscale platelet membranes (denoted as PV).

[0070] Add 1 mL of neoindocyanine green (4 mg / mL) to 1 mL of platelet membrane solution (protein content 5 mg / mL) and incubate at 4 °C for 12 h. Centrifuge at 20,000 rpm for 30 min to obtain a platelet membrane loaded with neoindocyanine green (denoted as IR-PV).

[0071] (d) Combine the 500 μL NP prepared above with 500 μL IR-PV (protein 1 mg / mL, IR: 200 μg), sonicate at 4 °C for 5 min, repeat twice with a 5 min interval, to obtain the photomagnetic visualization bio-hybrid nanosystem (denoted as IR-PV@NP).

[0072] Experimental Example

[0073] 1. The particle size distribution and zeta potential of Los NP, NP, LNP, IR-PV@NP, IR-PV@LNP and IR-PV were detected by a dynamic light scattering particle size analyzer.

[0074] Particle size distribution results are as follows Figure 1 As shown, the zeta potential analysis results are as follows: Figure 2 As shown;

[0075] Depend on Figure 1 , Figure 2 It can be known that:

[0076] The average particle size of Los NP is around 120 nm, the average particle size of NP is around 180 nm, the average particle size of LNP is around 200 nm, the average particle size of IR-PV@NP is around 220 nm, and the average particle size of IR-PV@NP is around 220 nm; all materials have a negative surface charge.

[0077] 2. Evaluation of the photothermal effect of nanoparticles:

[0078] The photothermal conversion efficiency of IR-PV@LNP was evaluated using EP tubes containing PBS, IR, and IR-PV@LNP (150 μg / ml). An 808 laser (1 W / cm²) was used. 2 Irradiate the EP tube and observe the temperature rise over 6 minutes using a thermal imager, then plot the curve; thermal imaging is as follows: Figure 3 As shown, the temperature change curve is as follows: Figure 4 As shown;

[0079] Depend on Figure 3 , Figure 4 It can be known that:

[0080] IR-PV@LNP has a photothermal conversion capability similar to that of free IR820, and can heat up to above 45.5℃ within 6 minutes. The excellent photothermal conversion performance of IR-PV@LNP provides strong support for effective anti-tumor photothermal therapy.

[0081] 3. Evaluation of the in vitro photothermal stability of nanoparticles:

[0082] Mix IR-PV@LNP (IR820: 150 μg / ml) in EP tubes. Then, use 1 W / cm². 2 After irradiating with an NIR laser for 5 minutes, the temperature changes over three irradiation cycles were detected using a thermal imager. The results are as follows: Figure 5 As shown;

[0083] Depend on Figure 5 It can be known that:

[0084] IR-PV@LNP can still reach above 45.5℃ after three irradiation cycles, demonstrating good photothermal conversion capability and photothermal stability.

[0085] 4. Evaluation of DC maturation induction and T cell proliferation:

[0086] 4T1 cells were first incubated with PBS, IR-PV@NP, and IR-PV@LNP (IR: 4 μg / ml, Los: 40 μg / ml, PV: 20 μg / ml) for 9 h. Immediately afterwards, the 4T1 cells were irradiated with a laser (808 nm, 1.0 W / cm²) for 5 min. Then, the supernatant and a small amount of tumor cells were collected and incubated with immature dendritic cells (DCs) isolated from BALB / c mice for 24 h. The DCs were stained and analyzed by flow cytometry.

[0087] Meanwhile, to further evaluate the proliferation efficiency of T cells, CD3... + T cells were incubated with DCs matured by the above stimulation at a ratio of 10:1 for 4 days. CD3 levels were then assessed using CCK-8. + T cell proliferation level.

[0088] Flow cytometry results as follows Figure 6 As shown, the T cell proliferation ratio is, for example... Figure 7 As shown;

[0089] Depend on Figure 6 , Figure 7 It can be known that:

[0090] IR-PV@LNP-mediated photothermal therapy can effectively activate adaptive immunity and induce in vitro DC maturation and T cell proliferation.

[0091] 5. CAF regulation:

[0092] First, 3T3 cells were activated with TGF-β at the optimal concentration of 75 ng / mL for 48 h. After activation, cells were treated with PBS, single Los NP, NP, LNP, or IR-PV@LNP in low serum medium (2% FBS) for 24 h. Immunofluorescence staining was performed to detect α-smooth muscle actin (α-SMA). These markers were monitored using CLSM. The results were analyzed using laser confocal microscopy. Figure 8 As shown;

[0093] Depend on Figure 8 It can be known that:

[0094] Under conditions where TGF-β activates normal mouse embryonic fibroblasts, causing them to highly express α-SMA, losartan can downregulate α-SMA expression. These results demonstrate that losartan can regulate CAF and reduce tumor extracellular matrix.

[0095] 6. In vitro tumor sphere permeability test:

[0096] The permeability of different drugs in tumor spheroids was analyzed to simulate drug diffusion in vivo. 96-well plates were prepared using agarose gel with a high gelation temperature (6×10⁻⁶). 3After overnight cell / well coating, 4T1 cells were seeded onto the bottom of an agarose gel and incubated for another 4 days to successfully prepare tumor spheres. The tumor spheres were then transferred to confocal culture dishes and treated with IR and IR-PV (IR: 4 μg / mL) for 12 h. Finally, the penetration of the drug into the tumor spheres was observed using a laser confocal microscope, and the results are as follows: Figure 9 As shown;

[0097] Depend on Figure 9 It can be known that:

[0098] Compared to IR, IR-PV has better tumor sphere penetration and can penetrate deeper from the edge of the tumor sphere.

[0099] 7. In vivo targeted delivery via photomagnetic imaging tracer:

[0100] To determine the tumor-targeting effect of IR-PV@LNP, tumor-bearing mice were intravenously injected with IR and IR-PV@LNP (IR: 1.1 mg / kg, Los: 22.2 mg / kg, PV: 5.6 mg / kg). Postoperative systemic drug accumulation was measured at different time points using a small animal in vivo imaging system. Figure 10 As shown.

[0101] Subsequently, the study aims to investigate the enhancement effect of IR-PV@LNP on in vivo tumor magnetic resonance imaging. This will be achieved in 4T1 tumor-bearing mice with a tumor volume reaching 100 mm². 3 Subsequently, mice were injected via tail vein with IR-PV@LNP (Mn: 3.14 mg / kg, IR: 1.1 mg / kg; Los: 22.2 mg / kg, PV: 5.6 mg / kg). Twenty-four hours after administration, sagittal and axial imaging were performed on the mice using a small animal magnetic resonance imaging system. The results are as follows: Figure 11 As shown.

[0102] Depend on Figure 10 It can be known that:

[0103] At the 24-hour time point, the accumulation of IR-PV@LNP in the tumor site was higher than that of free IR820, indicating that the delivery system has a good tumor targeting effect.

[0104] Depend on Figure 11 It can be known that:

[0105] T1-weighted magnetic resonance imaging (MRI) analysis of 4T1 orthotopic tumor-bearing mice injected with IR-PV@LNP via the tail vein revealed that, in the blank control group, the tumors in 4T1 mice exhibited low resolution on MRI images, making it difficult to determine their size and shape. However, after tail vein injection of IR-PV@LNP, the IR-PV@LNP accumulated at the tumor site, and the tumor outline became clearly discernible.

[0106] 8. In vivo tumor suppression effect study:

[0107] Tumor-bearing mice were randomly divided into five groups: G1, PBS; G2, IR-PV@NP; G3, IR-PV@LNP; G4, IR-PV@NP + laser; G5, IR-PV@LNP + laser. The start date of tumor implantation was recorded as day 0. On days 7 and 14, each group of mice was intravenously injected with a different formulation (IR: 1.1 mg / kg; Los: 22.2 mg / kg, PV: 5.6 mg / kg). On days 8 and 15, the tumor sites of the mice were irradiated with an 808 nm laser (1.0 W / cm², 5 min). Tumor volume was continuously measured every two days using electronic calipers and calculated as follows: Tumor volume = L × W × W / 2 (L, longest dimension; W, shortest dimension). Then, on day 25, the mice were sacrificed, and the tumors were photographed; the tumor growth curve is shown below. Figure 12 As shown, ex vivo tumors, such as Figure 13 As shown:

[0108] Depend on Figure 12 , 13 It can be known that:

[0109] IR-PV@LNP-mediated CAF regulation and photothermal-immunotherapy can effectively inhibit tumor growth.

[0110] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be covered within the scope of the claims and specification of the present invention.

Claims

1. A magneto-optically visualized bio-hybrid nanosystem, characterized in that, The light-magnetic visualized bio-hybrid nanosystem comprises a platelet membrane and a losartan-loaded tannic acid manganese complex wrapped in the platelet membrane; The platelet membrane further loads a new indocyanine green; The preparation method of the light-magnetic visualized bio-hybrid nanosystem comprises the following steps: (a) under ultrasonic conditions, a losartan solution is added dropwise into a polyvinylpyrrolidone solution and ultrasonic treatment is continued for a period of time to obtain a losartan nanoparticle solution, (b) a tannic acid solution and a manganese chloride solution are added dropwise into the losartan nanoparticle solution and ultrasonic treatment is performed, and then the pH value is adjusted to alkaline and ultrasonic treatment is continued, followed by centrifugation, washing and drying to obtain a losartan-loaded Mn-TA complex; (c) new indocyanine green is added into a platelet membrane solution for incubation and centrifugation to obtain a platelet membrane loading new indocyanine green; (d) the platelet membrane loading new indocyanine green is used to wrap the losartan-loaded Mn-TA complex by ultrasonic method to obtain the light-magnetic visualized bio-hybrid nanosystem.

2. The opto-magnetically visualized bio-hybrid nanosystem according to claim 1, characterized in that, In the step (a), the volume ratio of the losartan solution to the polyvinylpyrrolidone solution is 1:(15-25); The concentration of the losartan solution is 40-60 mg / ml; and the concentration of the polyvinylpyrrolidone solution is 0.8%-1.2%.

3. The opto-magnetically visualized bio-hybrid nanosystem according to claim 1, characterized in that, In the step (b), the volume ratio of the losartan nanoparticle solution to the tannic acid solution to the manganese chloride solution is (7-9):(2-4):(0.8-1.2). The concentration of the tannic acid solution is 8-12 mg / ml; and the concentration of the manganese chloride solution is 4-6 mg / ml.

4. The opto-magnetically visualized bio-hybrid nanosystem according to claim 1, characterized in that, In the step (b), the pH value is adjusted to 7.5-9.

5. The opto-magnetically visualized bio-hybrid nanosystem according to claim 1, wherein, In the step (c), the mass ratio of the protein content in the platelet membrane to the new indocyanine green is (2-3):

1.

6. The opto-magnetically visualized bio-hybrid nanosystem according to claim 1, wherein, In the step (c), the incubation temperature is 0-8℃, and the incubation time is 10-18 h.

7. The opto-magnetically visualized bio-hybrid nanosystem according to claim 1, wherein, In the step (d), the mass ratio of the protein content in the platelet membrane to the losartan is 1:(4-6).

8. Use of the light-magnetic visualized bio-hybrid nanosystem according to any one of claims 1-7 in the preparation of a product for treating breast cancer.

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