Probe for collaborative diagnosis and treatment monitoring as well as preparation method and application of probe

Abstract

The invention relates to the technical field of biomedicine, in particular to a probe for collaborative diagnosis and treatment monitoring and a preparation method and application thereof. The probe is a lipid nanoparticle, FTBGd and statins are taken as a hydrophobic core, and a phospholipid compound and poly (p-propiophenone ketene mercaptal) with good biocompatibility are selected as carriers for co-assembly, so that the probe is formed. According to the probe provided by the invention, the risk of phagocytosis by monocytes can be reduced, the circulation time of the probe in a body is prolonged, and targeted aggregation of atherosclerotic inflammatory endothelium is enhanced; multi-modal imaging of foam cells can be realized, so that early recognition and multi-dimensional information acquisition of atherosclerosis become possible; the expression of CD47 on the surface of foam cells can be reduced, the phagocytosis of macrophages can be enhanced, and the development of necrotic cores and plaques can be inhibited. The preparation method provided by the invention is mild in reaction condition, simple and convenient to operate and beneficial to large-scale production.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a probe for synergistic diagnosis and treatment monitoring, its preparation method, and its application. Background Technology

[0002] Despite significant progress in the diagnosis and prognosis of cardiovascular diseases over the past few decades, thanks to advancements in early detection methods, surgical techniques, and other advanced diagnostic and treatment approaches, cardiovascular disease remains the leading cause of death and disability worldwide. Atherosclerosis (AS), as the most common and clinically significant chronic inflammatory disease, is characterized by the progressive, asymptomatic accumulation of lipid-rich plaques in the arterial walls. As the disease progresses, these plaques may rupture, leading to adverse cardiovascular events such as myocardial infarction, stroke, and sudden death. Therefore, early detection and accurate monitoring of atherosclerosis are crucial for timely intervention, symptom relief, and halting disease progression. However, current diagnostic methods lack sufficient sensitivity and specificity in detecting early lesions, severely limiting the effectiveness of clinical management.

[0003] Currently used imaging techniques each have their inherent advantages, disadvantages, and applicable ranges. For example, intravascular ultrasound and optical coherence tomography (OCT) suffer from drawbacks such as low sensitivity, weak functional signals, limited imaging depth, and invasiveness. Magnetic resonance imaging (MRI), with its non-invasive and non-ionizing radiation characteristics, has become an ideal alternative for detecting atherosclerotic plaques. However, traditional small-molecule MRI contrast agents, including gadolinium compounds, suffer from low specificity, rapid systemic clearance, and poor image quality, severely affecting diagnostic accuracy. Optical imaging, renowned for its high sensitivity, real-time performance, cost-effectiveness, and ease of use, has become an important supplement to MRI, especially near-infrared II fluorescence imaging (FLI), which reduces tissue scattering and autofluorescence, resulting in superior tissue penetration and a higher signal-to-noise ratio. Furthermore, photoacoustic tomography (PAI), as a rapidly developing non-invasive imaging technique, offers excellent spatial resolution and tissue penetration, but suffers from lower sensitivity.

[0004] Therefore, single-molecule probes with integrated multi-imaging capabilities show great potential and offer significant advantages over traditional hybrid-mode imaging systems. However, optimizing the performance of each imaging modality at the single-molecule level presents a major challenge because interference often exists between different imaging components, hindering the selective differentiation between normal and pathological tissues. In view of this, given the complex heterogeneity of the pathological microenvironment in atherosclerosis, there is an urgent need to develop multimodal imaging strategies that integrate the advantages of various imaging modalities to improve the sensitivity and accuracy of atherosclerosis diagnosis.

[0005] Foam cells and the inflammatory responses they trigger play a crucial role in the pathogenesis of atherosclerosis (AS). These lipid-rich macrophages accumulate within atherosclerotic plaques, significantly exacerbating plaque instability and promoting plaque progression. Pathologically, foam cells are a key driver of plaque necrosis core formation; furthermore, as producers of potent pro-inflammatory cytokines, they recruit immune cells to the plaque site, amplifying the inflammatory cascade. In addition, the expression of CD47 and other molecules on the foam cell surface mediates a "do not phagocytose" signal, helping foam cells evade phagocytosis and clearance by the immune system.

[0006] Given the crucial role of foam cells in disease progression, precise imaging and effective regulation of them are essential for accurate detection and effective treatment of atherosclerosis. However, the complex microenvironment of atherosclerotic plaques presents challenges to targeted imaging and effective regulation of foam cells. Traditional imaging agents and drugs often suffer from low plaque accumulation efficiency, difficulty in penetrating the plaque core, and poor targeting of foam cells. These shortcomings not only affect imaging results but may also cause adverse side effects. Ultimately, these limitations severely undermine the accuracy of diagnostic strategies and the effectiveness of treatment strategies in the management of atherosclerosis.

[0007] Therefore, there is an urgent need to develop advanced multi-level targeting strategies to achieve precise imaging of pathological foam cells and lay the foundation for efficient therapeutic interventions. Summary of the Invention

[0008] This invention aims to at least solve one of the technical problems existing in related technologies. Therefore, the first objective of this invention is to provide a method for preparing a probe for synergistic diagnosis and treatment monitoring; the second objective is to provide a probe for synergistic diagnosis and treatment monitoring; and the third objective is to provide an application of the probe for synergistic diagnosis and treatment monitoring.

[0009] To achieve the first objective, the technical solution adopted by this invention is as follows: A method for preparing a probe for synergistic diagnosis and treatment monitoring includes the following steps: S100, Prepare an organic solution containing FTBGd, statins, phospholipids and poly(p-phenyleneacetone) dimethylene thioketal (PPADT). The structural formula for FTBGd is shown below: ; S200. Under stirring conditions, water is added to the organic solution to obtain lipid nanoparticles; S300. Using the lipid nanoparticles, a probe for synergistic diagnosis and treatment monitoring is prepared; The FTBGd structure uses the strongly electron-withdrawing benzobisthiadiazole group as the parent core, fluorene with strong fluorescence as the electron-donating group, and thiophene, which increases molecular conjugation and rotation, as a bridging unit, forming the "DAD" molecular structure. Here, D represents the donor and A represents the acceptor. The "DAD" molecular structure contains two donor D units and one acceptor A unit, which are covalently linked together. This molecular structure enhances the molecule's light absorption and photothermal conversion properties in the near-infrared region, ensuring both long-wavelength fluorescence emission and strong photoacoustic properties.

[0010] FTBGd possesses multiple imaging capabilities, enabling MRI, near-infrared II zone FLI, and PAI imaging, thereby improving upon the problems of component mixing, channel crosstalk, and poor repeatability in traditional multimodal imaging. By integrating these imaging modalities, a multifunctional platform for comprehensively assessing atherosclerosis can be constructed, aiming to guide subsequent treatment interventions and achieve in-situ evaluation of treatment effects.

[0011] This invention uses hydrophobic multimodal FTBGd and statin drugs as the core, and selects biocompatible phospholipid compounds and poly(p-phenylacetone ethoxylate) as carriers to encapsulate FTBGd and statin drugs, and co-assembles them to form a nanoprobe, which can realize precise multimodal imaging collaborative diagnosis and treatment monitoring of AS.

[0012] Furthermore, the preparation of FTBGd includes the following steps: S110, using compound II and compound IV Synthetic compound V ; S120. Using compound V, prepare compound VI. ; S130. Using compound VI, prepare compound VII. ; S140, Utilizing compound VII and compound The reaction was used to prepare compound VIII. ; S150. Using compound VIII, prepare FTBGd.

[0013] Furthermore, in step S110, the synthetic route for compound II is as follows: Pd(PPh3)4 is tetrakis(triphenylphosphine)palladium, NBS is N-bromosuccinimide, HBr is hydrobromic acid, and HCl is hydrochloric acid; The synthetic route for compound IV is shown below: .

[0014] Further, in step S100, the statin drug is selected from atorvastatin; The phospholipid compound is selected from distearate phosphatidylethanolamine-polyethylene glycol (DSPE-PEG). The solvent of the organic solution is selected from dimethyl sulfoxide.

[0015] Further, in step S100, the mass ratio of FTBGd, statin drugs, phospholipid compounds and poly(p-phenylacetone) thioglycol is 1:1:2:4.

[0016] Further, in step S200, the encapsulation efficiency of FTBGd in the lipid nanoparticles is not less than 63%, and the encapsulation efficiency of phospholipid compounds in the lipid nanoparticles is not less than 59%.

[0017] Furthermore, step S300 also includes the following steps: S310. The lipid nanoparticles are mixed with macrophage membrane material to prepare lipid nanoparticles with macrophage membrane modified on the surface. The preparation process of the macrophage cell membrane material is as follows: RAW264.7 cells are incubated in buffer for 12-24 hours and then ground. The ground cells are centrifuged at 20000g-50000g for 20-30 minutes, and the supernatant is collected. The collected supernatant is then centrifuged at 80000g-12000g for 50-70 minutes, and the supernatant is discarded to obtain the macrophage cell membrane material. Modifying the surface of lipid nanoparticles with biomimetic macrophage membranes can prolong the circulation time of the nanosystem in vivo and target adhesion to damaged endothelium at plaque sites, reduce non-specific clearance by the organism, and improve diagnostic accuracy and therapeutic monitoring.

[0018] Furthermore, step S310 also includes the following steps: S311. Lipid nanoparticles with macrophage membrane-modified surfaces are mixed with distearate phosphatidylethanolamine-polyethylene glycol-CLIKKPF to prepare lipid nanoparticles with targeted molecule-modified surfaces and macrophage membrane-modified surfaces. Lipid nanoparticles with surface-modified targeting molecules have active targeting properties for foam cells at the lesion site, which can improve the retention and enrichment of effective active ingredients at the lesion site.

[0019] To achieve the second objective, the technical solution adopted by this invention is as follows: A probe for collaborative diagnosis and treatment monitoring is prepared using any one of the above-described methods for preparing a probe for collaborative diagnosis and treatment monitoring. Among them, FTBGd serves as a synergistic diagnostic active ingredient, while statins serve as a therapeutic active ingredient.

[0020] To achieve the third objective, the technical solution adopted by this invention is as follows: Application of a probe for synergistic diagnosis and treatment monitoring: using the above-mentioned probe for synergistic diagnosis and treatment monitoring, to prepare medical devices or pharmaceutical preparations for the diagnosis and treatment of atherosclerosis. The diagnosis includes any one or more of the following: magnetic resonance (MR) imaging, fluorescence (FL) imaging, and photoacoustic (PA) imaging at the lesion site; The treatment includes one or more of the following: It has anti-inflammatory, lipid-lowering, and phagocytic effects on macrophages at the lesion site, and inhibits plaque formation and progression.

[0021] The above-described one or more technical solutions in the embodiments of the present invention have at least one of the following technical effects: This invention provides a probe for collaborative diagnosis and treatment monitoring, its preparation method, and its application. The probe is a lipid nanoparticle, co-assembled with FTBGd (capable of multimodal imaging) and statins as a hydrophobic core, and biocompatible phospholipids and poly(p-phenylacetone thioglycolate) as carriers. This enables precise multimodal imaging for collaborative diagnosis and treatment monitoring of atherosclerosis (AS). Experimental results show that the probe provides reduced risk of phagocytosis by monocytes, prolonged its circulation time in the body, and enhanced targeted aggregation of inflammatory endothelium in AS. Simultaneously, the probe can achieve MRI, FLI, and PAI three-mode imaging of foam cells. The organic combination and complementary advantages of these three imaging modes enable early identification and multidimensional information acquisition of atherosclerosis. Furthermore, the released pharmacologically active ingredient, statins, promotes lipid efflux from foam cells and reduces CD47 expression on their surface, thereby enhancing macrophage clearance of foam cells and inhibiting necrotic core and plaque progression.

[0022] Meanwhile, this invention provides a method for preparing a probe for synergistic diagnosis and treatment monitoring, which has mild reaction conditions, is easy to operate, and is conducive to large-scale production.

[0023] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0024] Figure 1 The results are molecular characterization results of FTBGd provided in Example 1 of this invention.

[0025] Figure 2 This is a diagram showing the near-infrared FL and MRI signal stability monitoring of FA-P@MC after different material treatments provided in Embodiment 2 of the present invention.

[0026] Figure 3 These are the measurement results from transmission electron microscopy (TEM) and dynamic light scattering (DLS) provided in Embodiment 2 of the present invention.

[0027] Figure 4 The optical property characterization results of FA-P@MC provided in Embodiment 2 of the present invention are shown.

[0028] Figure 5 This is a diagram showing the uptake of different lipid nanoparticles by foam cells, provided in Experimental Example 1 of this invention.

[0029] Figure 6 This is a confocal image of the in vitro filter migration experiment provided in Experiment Example 1 of the present invention.

[0030] Figure 7 This is a flow cytometry analysis diagram of the in vitro transfer filter migration experiment provided in Experiment Example 1 of the present invention.

[0031] Figure 8 This is a quantitative statistical bar chart of foam cells taking up different components in an in vitro transfiltration migration experiment provided in Experiment Example 1 of the present invention.

[0032] Figure 9 This is a diagram showing the results of detecting foam cells using the ROS fluorescent probe 2′,7′-dichlorofluorescein diacetate (DCFH-DA) provided in Experimental Example 2 of this invention.

[0033] Figure 10 This is a graph showing the evaluation results of the ability of FA-P@MC to reduce oxidized low-density lipoprotein uptake and inhibit foam cell formation provided in Experimental Example 2 of this invention.

[0034] Figure 11 This is a graph showing the results of different nanoparticles regulating CD47 expression, as provided in Experimental Example 3 of this invention.

[0035] Figure 12 This is a fluorescence imaging image of bone marrow-derived macrophages (BMDMs) and foam cells after different nanoparticle treatments provided in Experimental Example 3 of this invention.

[0036] Figure 13 This is a flow cytometry analysis diagram of foam cells being phagocytosed by phagocytes after different nanoparticle treatments provided in Experimental Example 3 of this invention.

[0037] Figure 14 This is provided in Experimental Example 4 of the present invention, which involves targeting apolipoprotein E-deficient (ApoE) proteins. — / — The results of evaluating the ability of FA-P@MC to detect plaques and differentiate plaque severity in mice are shown in the figure.

[0038] Figure 15 This is a graph showing the results of verifying the correlation between optical signal intensity and patch severity provided in Experiment Example 4 of this invention.

[0039] Figure 16 This is a result of verifying the ability of FA-P@MC multimodal imaging to detect plaques in living tissue, as provided in Experimental Example 5 of this invention.

[0040] Figure 17 This describes the in vivo efficacy of FA-P@MC in treating atherosclerosis (AS) provided in Experimental Example 6 of this invention.

[0041] Figure 18 This is a fluorescence imaging image of the carotid artery of an atherosclerotic mouse provided in Experimental Example 7 of this invention.

[0042] Figure 19 This is a photoacoustic (PA) imaging image of the carotid artery of an atherosclerotic mouse provided in Experimental Example 7 of this invention.

[0043] Figure 20 This is a graph showing the results of the safety assessment of FA-P@MC provided in Experiment Example 8 of this invention. Detailed Implementation

[0044] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below in conjunction with specific embodiments. Obviously, the described embodiments are only some embodiments of this invention, not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention. The following embodiments are used to illustrate this invention, but cannot be used to limit the scope of this invention.

[0045] A method for preparing a probe for synergistic diagnosis and treatment monitoring includes the following steps: S100, Prepare an organic solution containing FTBGd, statins, phospholipids and poly(p-phenylacetone) thioglycol; The structural formula for FTBGd is shown below: ; S200. Under stirring conditions, water is added to the organic solution to obtain lipid nanoparticles; S300. Using the lipid nanoparticles, a probe for synergistic diagnosis and treatment monitoring is prepared.

[0046] According to a specific embodiment of the present invention, step S300 further includes the following steps: S310. The lipid nanoparticles are mixed with macrophage membrane material to prepare lipid nanoparticles with macrophage membrane modified on the surface.

[0047] According to a specific embodiment of the present invention, step S310 further includes the following steps: S311. Lipid nanoparticles with macrophage membrane-modified surfaces are mixed with distearate phosphatidylethanolamine-polyethylene glycol-CLIKKPF to prepare lipid nanoparticles with macrophage membrane-modified surfaces and targeted molecule-modified surfaces.

[0048] In the following embodiments, unless otherwise specified, the experimental methods used are conventional methods, and the materials and reagents used are commercially available, unless otherwise specified, and are carried out in accordance with the techniques or conditions described in the literature in this field or in accordance with the product instructions.

[0049] Example 1: Preparation of FTBGd.

[0050] The structural formula for FTBGd is: .

[0051] I. Preparation of Compound II The synthetic route is shown below: Its synthesis process is as follows: Under an argon atmosphere, 3,4-dimethoxythiophene (2 g, 13.87 mmol) was dissolved in anhydrous tetrahydrofuran (THF) (20 mL), cooled to -78 °C, and maintained for 30 min. Then, a 1.6 M n-butyllithium (n-BuLi) hexane solution (954 μL, 15.26 mmol) was added, and the reaction was stirred for another 2 h. Tributyltin chloride (4.5 mL, 16.64 mmol) was then added and mixed thoroughly. The mixture was heated to room temperature and stirred for approximately 12 h. Water was added to quench the reaction, and the mixture was then extracted three times with dichloromethane. The collected organic phase was dried over MgSO4, and the solvent was removed under reduced pressure to obtain... (1.77 g, 4.62 mmol); it was added to anhydrous THF (20 mL) along with Pd(PPh3)4 (0.24 g, 0.04 mmol), heated under argon protection, stirred under reflux for 24 h, cooled to room temperature, water was added, and the mixture was extracted three times with dichloromethane (200 mL). The collected organic phases were mixed, dried over anhydrous MgSO4, concentrated, and the crude product was obtained. The crude product was purified by silica gel column chromatography (eluent was a mixture of dichloromethane and n-hexane, volume ratio 1:5) to give a purple solid compound I in 71% yield. 1 H NMR and 13 The characterization data of C NMR are as follows: 1 H NMR (400MHz, CDCl3): δ 6.58 (s, 2H), 3.90 (s, 6H), 3.85 (s, 6H); 13 C NMR (101MHz, CDCl3): δ 152.91, 150.07, 146.46, 121.57, 110.55, 100.90, 59.98, 57.52.

[0052] Under an argon atmosphere, compound I (3 g, 5.88 mmol) was dissolved in a mixed solvent of dimethylformamide (DMF) (30 mL) and acetonitrile (15 mL). The mixture was heated to 65 °C, and then NBS (2.09 g, 11.76 mmol) and 12 drops of HBr were slowly added. The mixture was stirred for 3 h in the dark, cooled to room temperature, and then 180 mL of 2 M hydrochloric acid solution was added. The mixture was stirred for 2 h, filtered, and the resulting filter cake was washed three times each with water (200 mL) and methanol (200 mL) to obtain a purple-red solid compound II, with a yield of 80%. 1 H NMR and 13 The characterization data of C NMR are as follows: 1 H NMR (400MHz, CDCl3): δ 3.97 (s, 6H), 3.85 (s, 6H); 13 C NMR (101MHz, CDCl3): δ 152.53, 149.65, 147.96, 120.33, 110.39, 103.61, 61.26, 60.06.

[0053] II. Preparation of Compound IV The synthetic route is shown below: ; Its synthesis process is as follows: 2-Bromofluorene (2 g, 8.16 mmol) and tert-butylacrylate (1.05 g, 8.16 mmol) were dissolved in toluene (100 ml). The mixture was heated to 120 °C under nitrogen protection and refluxed for about 12 h. After cooling to room temperature and terminating the reaction with water, the mixture was extracted three times with dichloromethane (100 ml). The combined organic phases were dried over anhydrous MgSO4 and concentrated to obtain a crude product. This crude product was purified by silica gel column chromatography (eluting solvent was a mixture of dichloromethane and hexane, volume ratio 1:2) to obtain compound III in 67% yield. 1 H NMR and 13 The characterization data of C NMR are as follows: 1 H NMR (400MHz, CDCl3): δ 7.66 (q, J=3.8Hz, 1H), 7.55 (d, J=8.0Hz, 1H), 7.51–7.45 (m, 2H), 7.35 (q, J=5.1, 4.3Hz, 3H), 2.39–2.25 (m, 4H), 1.50–1.42 (m, 4H), 1.31 (s, 18H); 13 C NMR (101MHz, CDCl3): δ 172.53, 150.34, 147.77, 140.18, 130.78, 128.12, 127.74, 126.42, 123.13, 121.54, 121.33, 120.07, 80.24, 53.79, 34.54, 29.95, 28.00.

[0054] Compound III (3.27 g, 6.52 mmol) and bis(pinacol borate) (4.2 g, 16.54 mmol) were dissolved in DMF (30 ml). Then, a dichloromethane solution (20 ml) containing potassium acetate (2.7 g, 27.51 mmol) and catalyst Pd(dppf)₂Cl₂ (103 mg, 0.125 mmol) was added. Under an argon atmosphere, the mixture was heated to 90 °C and refluxed with stirring for approximately 12 h. The mixture was then cooled to room temperature, and the reaction was terminated by adding water. The mixture was washed three times with dichloromethane (100 ml), and the combined organic phases were dried over anhydrous MgSO₄ and concentrated to obtain a crude product. This crude product was purified by silica gel column chromatography (using a mixed solvent of dichloromethane and hexane in a volume ratio of 2:1) to obtain a white solid compound IV in 85% yield. 1 H NMR and 13 The characterization data of C NMR are as follows: 1 H NMR (400MHz, CDCl3): δ 7.75 (d, J=7.5Hz, 1H), 7.70 (s, 1H), 7.62 (d, J=7.9Hz, 2H), 7.33–7.25 (m, 3H), 2.30 (q, J=5.7Hz, 4H), 1.38 (dd, J=11.5, 7.4Hz, 4H), 1.32 (s, 12H), 1.23 (s, 18H); 13 C NMR (101MHz, CDCl3): δ 172.89, 148.72, 147.30, 144.07, 140.97, 134.39, 128.98, 128.16, 127.49, 123.15, 120.41, 119.29, 83.88, 80.00, 53.47, 34.57, 30.00, 28.01, 24.95.

[0055] III. Preparation of compound V, the synthetic route is shown below: Its preparation process is as follows: Compound II (307.4 mg, 0.46 mmol) and compound IV (500 mg, 0.91 mmol) were dissolved in toluene (30 mL), and 2.5 M potassium carbonate aqueous solution (201.7 mg, 1.46 mmol) and catalyst Pd(PPh3)4 (10.6 mg, 0.0092 mmol) were added. The mixture was heated to 110 °C and refluxed for 24 h under nitrogen protection. After the reaction was completed, the mixture was cooled to room temperature, and water was added. The mixture was then extracted three times with dichloromethane (100 mL). The organic phases were combined, dried over anhydrous MgSO4, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (using a mixture of dichloromethane and hexane as eluent, with a volume ratio of 5:1) to obtain a purple solid compound in 76% yield. 1 H NMR and 13 The characterization data of C NMR are as follows: 1 H NMR (400MHz, CDCl3): δ 7.83 (d, J=1.6Hz, 2H), 7.81–7.72 (m, 6H), 7.41–7.35 (m, 6H), 3.95 (s, 6H), 3.84 (s, 6H), 2.40 (dd, J=9.8, 7.0Hz, 8H), 1.58 (s, 8H), 1.32 (s, 36H); 13 C NMR (101MHz, CDCl3): δ 172.69, 148.46, 128.10, 127.72, 126.63, 123.17, 121.49, 120.41, 120.25, 80.20, 60.65, 53.64, 34.69, 30.10, 28.02.

[0056] IV. Preparation of compound VI, the synthetic route of which is shown below: Its preparation process is as follows: Compound V (30 mg, 0.022 mmol) and ammonium chloride (42.26 mg, 0.79 mmol) were dissolved in a mixed solvent containing dichloromethane (7.2 mL), methanol (10.26 mL), and water (1.14 mL). Zinc powder (171.6 mg, 2.64 mmol) was added. The mixture was stirred for 4 h at room temperature under a nitrogen atmosphere. The reaction was terminated by adding water. The reaction solution was washed three times with dichloromethane (100 mL). The organic phases were combined, dried over anhydrous MgSO4, and concentrated to obtain intermediate I. This intermediate, benzenesulfonamide (18.4 mg, 0.132 mmol), and trimethylchlorosilane (21.5 mg, 0.198 mmol) were added to anhydrous pyridine (10 mL), and the mixture was heated to 80 °C under nitrogen protection. The mixture was refluxed at ℃ for approximately 12 hours, cooled to room temperature, dried, and then water was added to terminate the reaction. The reaction solution was extracted three times with dichloromethane (100 ml). The combined organic phases were dried over anhydrous MgSO4 and concentrated to obtain a crude product. This crude product was purified by silica gel column chromatography (eluting solvent was a mixture of dichloromethane and methanol, volume ratio 100:1) to obtain a green solid compound VI in 47% yield. 1 H NMR, 13 The characterization data of C NMR and MALDI-TOF are as follows: 1 H NMR (400MHz, CDCl3): δ 7.97 (d, J=8.0Hz, 1H), 7.90 (s, 1H), 7.80 (d, J=6.7Hz, 2H), 7.71–7.64 (m, 4H), 7.35–7.28 (m, 6H), 4.01–3.83 (m, 12H), 2.34 (q, J=7.1Hz, 8H), 1.52–1.45 (m, 8H), 1.25 (s, 36H); 13 C NMR (101MHz, CDCl3): δ 172.76, 153.63, 148.77, 148.42, 140.68, 127.89, 127.69, 126.72, 126.46, 123.13, 121.3 4, 120.37, 120.12, 80.19, 80.16, 60.47, 60.24, 53.65, 53.57, 34.72, 30.11, 28.03, 14.16; MALDI-TOF: C 72 H 78 N4O 12 S4, calculated value is 1318.4499, measured value is 1318.4526.

[0057] IV. Preparation of compound VII, the synthetic route is shown below: ; Its preparation process is as follows: Compound VI (148 mg, 0.11 mmol) was dissolved in dichloromethane (15 mL), cooled to 0 °C, and trifluoroacetic acid (8 mL) was added. The mixture was stirred for 3 h under a nitrogen atmosphere, slowly heated to room temperature, and the solvent was removed under vacuum. The solution was washed three times with dichloromethane (100 mL) to obtain a dark black solid compound VII, with a yield of 79%. Its high-resolution mass spectrometry (HRMS) characterization data are as follows: HRMS (ESI): C 56 H 46 N4O 12 S4[M], calculated value is 1094.20, measured value is 1094.16.

[0058] V. Preparation of compound VIII, the synthetic route is shown below: Its preparation process is as follows: Compound VII (20 mg, 0.018 mmol) and (22.1 mg, 0.036 mmol) was dissolved in DMF (5 mL), followed by the addition of o-benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate (27.3 mL, 0.072 mmol) and N,N-diisopropylethylamine (33.4 mg, 0.258 mmol). The reaction was carried out under nitrogen atmosphere at room temperature for about 12 h, and then water was added to terminate the reaction. The reaction solution was washed three times with dichloromethane (100 mL), and the combined organic phases were dried over anhydrous MgSO4 and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (eluting solvent was a mixture of dichloromethane and methanol, with a volume ratio of 5:1) to obtain product VIII, with a yield of 72%.

[0059] VI. Preparation of FTBGd, the synthetic route is shown below: Its synthesis process is as follows: Compound VIII (90 mg, 0.04 mmol) was dissolved in dichloromethane (15 mL), cooled to 0 °C, and trifluoroacetic acid (8 mL) was added. The mixture was stirred for 3 h under a nitrogen atmosphere, slowly heated to room temperature, and the solvent was removed under reduced pressure. The solution was washed three times with dichloromethane (100 mL) to obtain a dark black solid compound FTB. FTB was dissolved in methanol (25 mL), and the pH of the reaction solution was adjusted to approximately 5 with dilute sodium hydroxide solution. Anhydrous gadolinium chloride (16 mg, 0.06 mmol) was dissolved in methanol (3 mL) and added dropwise to the above reaction solution. The reaction was carried out at 60 °C for approximately 12 h. The methanol was evaporated to remove the crude product. The crude product was dissolved in dichloromethane, and excess gadolinium ions were washed away with water to obtain a black solid FTBGd with a yield of 56%. Its HRMS characterization data are as follows: HRMS (ESI): C 92 H 104 Gd2N 16 O 24 S4[MH], calculated value is 2261.4879, measured value is 2261.4836; Molecular characterization of FTBGd, such as Figure 1 As shown; Figure A shows the absorption and emission spectra of FTBGd in THF. Figure B shows the relationship between the fluorescence peak intensity of FTB and FTBGd and the water content in a mixed solvent of THF and H2O. I 0 and I The values ​​represent the FL strength in pure THF and mixed solvents with different water contents, respectively. Figure C shows the PA spectrum of FTBGd; Figure D is a bar chart showing the quantitative analysis of the magnetic resonance signal intensity of FTB, Gd-DOTA, and FTBGd. Gd-DOTA is a core component of a commonly used clinical MRI contrast agent, consisting of gadolinium ions (Gd). 3+ The stable chelate formed by the DOTA ligand is shown in the following structural formula: .

[0060] Example 2 The process for preparing probes for synergistic diagnosis and treatment monitoring is as follows: At room temperature, FTBGd (1 mg), atorvastatin (AT) (1 mg), DSPE-PEG (2 mg), and PPADT (4 mg) were dissolved in dimethyl sulfoxide (1 mL) under continuous stirring to obtain an organic solution. This organic solution was then slowly added to deionized water (15 mL) under stirring. DSPE-PEG was introduced into the system as a surfactant during the formation of lipid nanoparticles. Due to its amphiphilic structure, DSPE-PEG anchors the hydrophobic lipid tails to the core of the lipid nanoparticles, while simultaneously forming a hydrophilic PEG crown on the surface, thereby enhancing the stability of the lipid nanoparticles. To remove the organic solvent, the prepared lipid nanoparticles were dialyzed for 12 hours using a dialysis membrane with a molecular weight cutoff of 3500 Da to obtain a probe for synergistic diagnosis and treatment monitoring, named FA-P. In FA-P, the encapsulation efficiencies of FTBGd and AT were 63.7% and 59.5%, respectively.

[0061] FA-P (100 μL, based on an FTBGd concentration of 1 mg / mL) was mixed with macrophage membrane material (0.5 mg) and treated with sonication (100 W, 50 Hz) for 5 min. The mixture was then repeatedly extruded through a microextruder with a pore size of 200 nm to form lipid nanoparticles modified with macrophage membrane, denoted as FA-P@M.

[0062] The preparation process of the macrophage cell membrane material is as follows: RAW264.7 cells were resuspended in PBS to obtain a cell density of 3 × 10⁻⁶ cells / mL. 5 The cells were suspended in a cell suspension of 1 cell / ml and incubated overnight in hypotonic lysis buffer (4°C). The cells were then ground 20 times with a pestle and centrifuged at 20,000g for 25 min. The supernatant was collected and centrifuged at 100,000g for 60 min. The supernatant was discarded to obtain macrophage membrane material.

[0063] FA-P@M (0.2 mg) and DSPE-PEG-CLIKKPF (0.1 mg) were mixed and treated with sonication (100 W, 50 Hz) for 5 min. The mixture was then repeatedly extruded through a micro extruder with a pore size of 200 nm to obtain lipid nanoparticles with surface-targeted molecule modification and macrophage membrane modification, denoted as FA-P@MC. The preparation process of DSPE-PEG-CLIKKPF is as follows: At room temperature, CLIKKPF (0.15 mg) peptide and DSPE-PEG-maleimide (0.4 mg) are added to 15 ml of a mixed solvent of water and methanol (water to methanol volume ratio of 9:1), and reacted for 8 h. Then, residual CLIKKPF peptide is removed by centrifugation filter (Merck Millipore Ltd., Shanghai, China), and lyophilized to obtain DSPE-PEG-CLIKKPF.

[0064] The stability of near-infrared FL and MRI signals of FA-P@MC after treatment with different substances, such as Figure 2 As shown in the figure; the blank areas represent the near-infrared FL and MRI signals of FA-P@MC without any material treatment.

[0065] The measurement results of FA-P@MC under different conditions using transmission electron microscopy (TEM) and dynamic light scattering (DLS) are as follows: Figure 3 As shown; In the figure, Figure A is the TEM image of FA-P@MC, Figure B is the particle size distribution of FA-P@MC, Figure C is the TEM image of FA-P@MC in a PBS solution with a concentration of 1.0 mM hydrogen peroxide, and Figure D is the particle size distribution of FA-P@MC in a PBS solution with a concentration of 1.0 mM hydrogen peroxide. The PBS solution with a concentration of 1.0 mM hydrogen peroxide was set up to simulate the oxidative stress conditions of atherosclerotic plaques. As can be seen from the TEM image, after treatment with reactive oxygen species (ROS), the morphology of FA-P@MC changed to an irregular fragmented state. This indicates that FA-P@MC can degrade under ROS conditions, releasing the diagnostic and therapeutic active ingredients encapsulated within it.

[0066] The optical property characterization results of FA-P@MC, such as Figure 4 As shown; Figure A shows the absorption spectrum of FA-P@MC; Figure B shows the FL spectrum of FA-P@MC; Figure C compares the photostability of FA-P@MC with the clinical fluorescent indicator indocyanine green (ICG); Figure D shows the PA spectrum of FA-P@MC; Figure E shows the linear relationship between different concentrations of FA-P@MC and PA signals, and the PA stability of FA-P@MC; Figure F shows the magnetic resonance signal intensity maps of FA-P@MC1 (with FTB as the imaging component), Gd-DOTA, and FA-P@MC (with FTBGd as the imaging component).

[0067] Experimental Example 1 The uptake of three different lipid nanoparticles (FA-P, FA-P@M, and FA-P@MC) obtained in Example 2 by foam cells was evaluated as follows: FA-P, FA-P@M, and FA-P@MC were co-incubated with foam cells at 37 °C for 4 h, respectively. The foam cells were then visualized using a confocal laser scanning microscopy (CLSM). The results are as follows: Figure 5 As shown in Figure A, the fluorescence imaging diagram is shown, and Figure B is the fluorescence intensity quantitative analysis diagram. The results show that the uptake of FA-P@M by foam cells is significantly higher than that of FA-P without macrophage membrane. In addition, after introducing the PS targeting peptide (CLIKKPF) into FA-P@MC, the cell uptake increased by 4.1 times compared with FA-P and by 1.6 times compared with FA-P@M. The migration ability of three different lipid nanoparticles, FA-P, FA-P@M, and FA-P@MC, through inflammatory human umbilical vein endothelial cells (HUVECs) and their uptake by foam cells were evaluated using an in vitro transfiltration migration assay. The procedure is as follows: Using the Transwell chamber device, a co-culture system with an upper and lower chamber structure was constructed to recreate cell interactions in the vascular microenvironment; The upper chamber contains HUVECs cells to simulate vascular endothelium, while the lower chamber contains foam cells to simulate diseased cells. The upper chamber is treated with the following components: FA-P+LPS, FA-P@M+LPS, FA-P@MC+LPS, and FA-P@MC; LPS is a lipopolysaccharide that acts as an inflammation inducer. Confocal images of the uptake of different components by lower chamber foam cells, such as... Figure 6 As shown in the figure, the nanoparticle signal in the foam cells of the FA-P treatment group is weak, indicating that the passive transendothelial migration of unmodified nanoparticles is limited; the FA-P@MC group shows a significantly enhanced nanoparticle signal in the basal side compartment, which indicates that after being coated by the macrophage membrane, its endothelial permeability is enhanced and the foam cell uptake capacity is improved.

[0068] Flow cytometry analysis of the uptake of different components by lower chamber foam cells, such as... Figure 7As shown in the figure, the lipid nanoparticles (i.e., FA-P@MC) with surface modified by targeted molecules and macrophage membranes can significantly promote the nanoparticles to cross the inflammatory endothelial barrier and promote their uptake by foam cells.

[0069] Quantitative statistical bar chart of the uptake of different components by lower chamber foam cells. Figure 8 As shown in the figure, the number of lipid nanoparticles migrating across the endothelium into basal foam cells was 5.24 times and 1.88 times higher than that of the FA-P group and the FA-P@M group, respectively. Furthermore, in the absence of LPS pretreatment to induce an inflammatory state, the permeability of FA-P@MC was significantly reduced, indicating that it possesses enhanced endothelial penetration and foam cell uptake capacity under inflammatory conditions.

[0070] Experimental Example 2 To evaluate the therapeutic effect of FA-P@MC, an in vitro foam cell model was established using macrophages treated with oxidized low-density lipoprotein. The procedure is as follows: After incubation with oxidized low-density lipoprotein (OxLDL, 50 µg / mL) for 24 h, foam cells were analyzed using the ROS fluorescent probe 2′,7′-dichlorofluorescein diacetate (DCFH-DA) to observe the oxidative stress response to different substances. The results are as follows: Figure 9 As shown in the figure, FA-P@M and FA-P@MC treatments reduced the intensity of DCFFL, with the FA-P@MC treatment group showing the most significant effect. Subsequently, this invention evaluated the ability of FA-P@MC to reduce oxidized low-density lipoprotein uptake and inhibit foam cell formation. The procedure was as follows: RAW264.7 macrophages were treated with 50 µg / mL oxidized low-density lipoprotein for 24 h. Intracellular lipid droplet accumulation and foam cell formation were examined by Oil Red O staining. The results are as follows: Figure 10 As shown in the figure, compared with other groups, the FA-P@MC treatment group significantly inhibited the formation of foam cells.

[0071] Experimental Example 3 This invention investigated whether different nanoparticles could enhance phagocytosis by regulating CD47 expression, as described below: RAW264.7 cells were first incubated with 0xLDL (50 µg / mL) for 24 h, then divided into groups. Each group was treated with different nanoparticles, washed three times with PBS, fixed with 4% paraformaldehyde for 15 min, blocked with 5% bovine serum albumin (BSA) at room temperature for 1 h, incubated with anti-CD47 primary antibody at 4 °C for 1 h, washed three times with PBS, and then incubated with horseradish peroxidase (HRP)-labeled secondary antibody at room temperature for 1 h. The expression level of CD47 on the cell surface of different nanoparticle treatment groups was quantitatively analyzed using a laser confocal microscopy system. The results are as follows: Figure 11 As shown, this result indicates that FA-P@MC can significantly reduce the expression of CD47 on the surface of foam cells.

[0072] In the phagocytosis study, bone marrow-derived macrophages (BMDMs) labeled with carboxyfluorescein succinimide were used as phagocytic cells, and RAW264.7 cells treated with 50 µg / mL OxLDL were used as foam cells. Foam cells were labeled with dioctyl-tetramethylindocyanine dicarbocyanine dye. BMDMs and foam cells treated with different nanoparticles were then co-cultured at a volume ratio of 1:3 for 6 h. After the culture medium was discarded, the cells were washed with PBS and then fixed with 4% paraformaldehyde for 15 min. Images were acquired using a confocal laser scanning microscope. The FL imaging of BMDMs and foam cells treated with different nanoparticles is shown in the figure below. Figure 12 As shown, the results indicated that, compared with the PBS-treated group, the proportion of foam cells treated with FA-P@MC that were phagocytosed by bone marrow-derived macrophages was significantly higher. The cells were then washed with PBS to detach them from the wells and analyzed by flow cytometry. The flow cytometry analysis of the phagocytosis of foam cells treated with different nanoparticles is shown below. Figure 13 As shown, this result indicates that the phagocytic rate of foam cells in the FA-P@MC group (approximately 68%) is much higher than that in the PBS, FA-P, and FA-P@M groups.

[0073] Experiment Example 4 To determine whether FA-P@MC can serve as a reliable indicator for assessing plaque progression, the inventors first evaluated the ability of the FA-P@MC probe to detect plaques and differentiate plaque severity, as follows: By targeting apolipoprotein E deficiency (ApoE) — / — A mouse model of atherosclerosis was induced by feeding mice a high-fat diet for 16 weeks. The tissue analysis results are as follows: Figure 14 As shown in Figure A1; subsequently, FA-P@MC was injected intravenously. 12 hours later, the aorta was removed and fluorescence imaging was first performed using the NIR-II imaging system, as shown in Figure A1.Figure 14 As shown in Figure B1, different arterial regions exhibit significant signal differences. Based on NIR-II FL intensity, the aorta was divided into four regions, A to D. Subsequently, PA angiography and MR imaging were performed on each segment. The results were consistent with the NIR-II FL findings: both FL signal intensity and MRI signal intensity showed a progressive increase from segment A to segment D. Figure 14 As shown in Figures C1 and D1, these results confirm the consistency of signal intensity across the three imaging modalities.

[0074] To verify the correlation between optical signal intensity and plaque severity, these aortic segments were stained with ORO and hematoxylin-eosin (H&E). The results are as follows: Figure 15 As shown in Figure A1, this result indicates that no plaque formation was observed in segment A, while the plaque area in segments B, C, and D progressively increased, reflecting the progression of plaque severity. Based on the H&E staining images, key pathological parameters of the plaques in segments A to D were quantified, including the necrotic area and the proportion of the necrotic core to the total plaque area. The results are as follows: Figure 15 As shown in Figure B1, this result indicates that the necrotic area progressively expands from segment B to segment D, suggesting a continuous increase in plaque instability from segment B to segment D. Stronger signal intensity was detected in more severe and fragile plaque areas, highlighting the potential of the multimodal FA-P@MC nanoprobe in accurately delineating plaque lesions and as a non-invasive method for assessing plaque severity.

[0075] Experimental Example 5 The validation of FA-P@MC's ability to detect plaques in vivo using FL, PA, and MRI trimodal imaging is shown in the following results. Figure 16 As shown, the verification process is as follows: ApoE induced by a high-fat diet — / — A mouse model of atherosclerosis was established, and then mice were intravenously injected with equal doses of PBS, FA-P@M, or FA-P@MC to compare the imaging performance of different lipid nanoparticles. Figure 16 As shown in Figure A, in mice treated with FA-P@C (lipid nanoparticles with a surface modified with the targeting molecule CLIKKPF but without macrophage membrane modification), the carotid artery showed only weak fluorescence 12 hours after injection. In contrast, the lipid nanoparticles FA-P@M with a surface modified with a macrophage membrane exhibited enhanced plaque accumulation capacity. The lipid nanoparticles FA-P@MC with both the targeting molecule and macrophage membrane modifications significantly increased plaque accumulation, almost doubling that of the FA-P@M group. Imaging of FL in ex vivo carotid artery samples, such as... Figure 16As shown in Figure B, compared with FA-P@C and FA-P@M, FA-P@MC accumulated significantly more in the aortic root and thoracic aorta, a result consistent with in vivo experimental results. Twelve hours after administration of equivalent doses of FA-P@C, FA-P@M, or FA-P@MC, anatomical localization was assessed using two-dimensional PA / US imaging of the carotid artery (transverse and sagittal views). Results are as follows: Figure 16 As shown in Figure C, compared with the PBS control group, the carotid artery in the FA-P@C group showed only a weak fluorescence signal under 790 nm excitation, indicating that foam cell targeting alone is insufficient to achieve effective plaque localization. The FL signal was stronger in the carotid artery plaques of the FA-P@M treatment group, which is attributed to the enhanced plaque enrichment capacity of the macrophage membrane biomimetic material. Notably, the carotid artery plaques of the FA-P@MC treatment group showed a strong FL signal, indicating that the synergistic effect of macrophage membrane coating and foam cell-targeting peptide modification significantly improved the enrichment efficiency of lipid nanoparticles in the plaque. In the image, US represents ultrasound imaging; In addition to FL and PA imaging, nanoprobes embedded with gadolinium complexes can also be used for in vivo MRI detection, enabling comprehensive monitoring of atherosclerosis. Gd-DOTA, FA-P@M, or FA-P@MC can be injected into ApoE. — / — Imaging comparisons were performed in mice before and after the procedure, and the results are as follows: Figure 16 As shown in Figure D, 12 hours after FA-P@MC injection, the walls of both the left and right common carotid arteries exhibited significant heterogeneity compared to the pre-injection images. T The enhanced MRI signal indicates the presence of plaque formation in these areas.

[0076] Experimental Example 6 This invention further investigated the in vivo efficacy of FA-P@MC in treating atherosclerosis (AS), and the results are as follows: Figure 17 As shown, the process is as follows: First, ApoE — / — Mice were fed a high-fat diet for 8 weeks to induce the formation of atherosclerotic plaques at arterial bifurcation. The atherosclerotic mice were then randomly divided into six treatment groups: a PBS control group, a FA-P group, a FA-P@M group, and a FA-P@MC group. After treatment, the aorta was completely removed, and the efficacy of each treatment group in treating atherosclerotic plaques was evaluated by ORO staining of the aortic arch (this staining can reveal lipid deposition). The results are as follows: Figure 17As shown in Figure A, the PBS group exhibited the largest ORO-positive area (approximately 35.74%), indicating significant plaque formation. In the FA-P@M group, the plaque area was significantly reduced (approximately 11.82%). FA-P@MC treatment significantly reduced the ORO-positive plaque area to approximately 5.32%, resulting in the smallest ORO-stained lesions in the aorta. These results demonstrate that FA-P@MC possesses potent anti-atherosclerotic efficacy.

[0077] H&E staining results of aortic root plaques, such as Figure 17 As shown in Figure B, compared to other experimental groups, the FA-P@MC treatment group showed a significant reduction in both plaque volume and necrotic core area, demonstrating superior performance.

[0078] Immunofluorescence image results, such as Figure 17 As shown in Figure C, the PBS-treated control group exhibited high levels of CD47 in its plaques, allowing apoptotic foam cells to evade macrophage-mediated clearance. In contrast, FA-P@MC treatment significantly reduced CD47 expression levels within the plaques. Compared to other nanoparticles, FA-P@MC demonstrated stronger foam cell regulation capabilities, consistent with its enhanced affinity for atherosclerotic plaques and improved ability to target foam cells. These two properties jointly facilitated the targeted delivery of ROS-clearing polymers and AT, enabling them to enter plaque foam cells for synergistic therapy.

[0079] MMP-9 staining, Masson's trichrome staining, and α-smooth muscle actin (α-SMA) staining results are as follows: Figure 17 As shown in Figure D; As can be seen from the MMP-9 staining images: the significantly positive MMP-9 staining observed in the aortic root sections of the PBS group was different, and FA-P@MC administration led to a significant decrease in MMP-9 expression; Masson trichrome staining images show that the collagen content around the plaques in the FA-P@MC treatment group was significantly increased, and the fibrous cap was thickened, thereby enhancing the structural integrity of the plaques.

[0080] The α-smooth muscle actin (α-SMA) staining image shows that the number of VSMCs in FA-P@MC treated plaques is significantly increased, indicating that the fibrous cap structure is more robust and the plaque stability is improved.

[0081] Experimental Example 7 To assess the feasibility of FA-P@MC monitoring plaque progression after treatment, mice were intravenously injected with FA-P@MC, and NIR-II fluorescence imaging was performed 12 hours after injection. The results are as follows: Figure 18As shown in the figure, the scale bar is 1 cm. It can be seen from the figure that the carotid artery of the atherosclerotic mice in the PBS group showed a strong NIR-II fluorescence signal, indicating severe plaque development; while the carotid artery of the mice treated with FA-P@MC only showed a weak NIR-II FL signal, indicating significant plaque progression after treatment. To further elucidate the plaque progression mechanism, PA imaging was performed. PA imaging was conducted 12 hours after lipid nanoparticle injection under 790 nm laser excitation. The results are as follows: Figure 19 As shown in the figure, the scale bar is 500 μm. It can be seen from the figure that compared with the PBS control group, the aortic fluorescence signal of atherosclerotic mice in the FA-P@MC treatment group was significantly reduced. This PA imaging result is consistent with the NIR-II FL imaging data.

[0082] Experimental Example 8 To investigate the safety of FA-P@MC, paraffin sections of the heart, liver, spleen, lung, and kidney of mice in the PBS and FA-P@MC intravenous injection groups were prepared and pathologically stained. The results are as follows: Figure 20 As shown in A in the figure, it can be seen from the figure that the mice treated with FA-P@MC did not have obvious organ damage; The results of routine blood tests and blood biochemistry analyses are as follows: Figure 20 As shown in Figures B and C, the results indicate that no abnormalities were observed in the mice treated with FA-P@MC, suggesting that FA-P@MC has good drug safety.

[0083] 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 of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing a probe for synergistic diagnosis and treatment monitoring, characterized in that, Includes the following steps: S100, Prepare an organic solution containing FTBGd, statins, phospholipids and poly(p-phenylacetone) thioglycol; The structural formula for FTBGd is shown below: ； S200. Under stirring conditions, water is added to the organic solution to obtain lipid nanoparticles; S300. Using the lipid nanoparticles, a probe for synergistic diagnosis and treatment monitoring is prepared.

2. The method for preparing the probe for collaborative diagnosis and treatment monitoring as described in claim 1, characterized in that, The preparation of FTBGd includes the following steps: S110, using compound II and compound IV Synthetic compound V ; S120. Using compound V, prepare compound VI. ; S130. Using compound VI, prepare compound VII. ; S140, Utilizing compound VII and compound The reaction was used to prepare compound VIII. ; S150. Using compound VIII, prepare FTBGd.

3. The method for preparing the probe for collaborative diagnosis and treatment monitoring as described in claim 2, characterized in that, In step S110, the synthetic route for compound II is shown below: Pd(PPh3)4 is tetrakis(triphenylphosphine)palladium, NBS is N-bromosuccinimide, HBr is hydrobromic acid, and HCl is hydrochloric acid; The synthetic route for compound IV is shown below: 。 4. The method for preparing the probe for collaborative diagnosis and treatment monitoring as described in claim 1, characterized in that, In step S100, the statin drug is selected from atorvastatin; The phospholipid compound is selected from distearate phosphatidylethanolamine-polyethylene glycol; The solvent of the organic solution is selected from dimethyl sulfoxide.

5. The method for preparing the probe for collaborative diagnosis and treatment monitoring as described in claim 1, characterized in that, In step S100, the mass ratio of FTBGd, statin, phospholipid compound and poly(p-phenylacetone ketene thioglycolate) is 1:1:2:

4.

6. The method for preparing the probe for collaborative diagnosis and treatment monitoring as described in claim 1, characterized in that, In step S200, the encapsulation efficiency of FTBGd in the lipid nanoparticles is not less than 63%, and the encapsulation efficiency of phospholipid compounds in the lipid nanoparticles is not less than 59%.

7. The method for preparing the probe for collaborative diagnosis and treatment monitoring as described in claim 1, characterized in that, Step S300 also includes the following steps: S310. The lipid nanoparticles are mixed with macrophage membrane material to prepare lipid nanoparticles with macrophage membrane modified on the surface. The preparation process of the macrophage membrane material is as follows: RAW264.7 cells are incubated in buffer for 12-24 hours and then ground. The ground cells are centrifuged at 20,000g-50,000g for 20-30 minutes, and the supernatant is collected. The collected supernatant is then centrifuged at 80,000g-12,000g for 50-70 minutes, and the supernatant is discarded to obtain the macrophage membrane material.

8. The method for preparing the probe for collaborative diagnosis and treatment monitoring as described in claim 7, characterized in that, Step S310 also includes the following steps: S311. Lipid nanoparticles with macrophage membrane-modified surfaces are mixed with distearate phosphatidylethanolamine-polyethylene glycol-CLIKKPF to prepare lipid nanoparticles with both target molecule-modified and macrophage membrane-modified surfaces.

9. A probe for collaborative diagnosis and treatment monitoring, characterized in that, It is prepared using the method for preparing a probe for synergistic diagnosis and treatment monitoring as described in any one of claims 1 to 8; Among them, FTBGd serves as a synergistic diagnostic active ingredient, while statins serve as a therapeutic active ingredient.

10. The application of a probe for collaborative diagnosis and treatment monitoring, characterized in that, Using the probe for synergistic diagnosis and treatment monitoring as described in claim 9, a medical device or pharmaceutical preparation for the diagnosis and treatment of atherosclerosis is prepared. The diagnosis includes any one or more of the following: magnetic resonance imaging, fluorescence imaging, and photoacoustic imaging at the lesion site; The treatment includes one or more of the following: It has anti-inflammatory, lipid-lowering, and phagocytic effects on macrophages at the lesion site, and inhibits plaque formation and progression.

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