Near-infrared two-region nanoprobe HS-1-coated PLGA (poly (lactic-co-glycolic acid)) as well as preparation method and application thereof

Abstract

The invention relates to a near-infrared two-region nanoprobe HS-1-coated PLGA, a preparation method thereof and application of the near-infrared two-region nanoprobe HS-1-coated PLGA in renal injury detection, and belongs to the technical field of biological medicines. The nanoprobe is formed by self-assembly of an HS-1 fluorescent dye, a polylactic acid-glycolic acid copolymer (PLGA), lecithin (Lecthin) and DSPE-PEG, and has good colloidal stability and fluorescence performance. The probe can be quickly distributed in vivo and enriched in a kidney area after intravenous injection administration, and high-contrast imaging is realized under a near-infrared second-area wave band. The preparation method is simple and efficient, imaging signals can be clearly presented within 30 minutes, and the method is suitable for imaging of various kidney injuries. Animal experiments prove that the probe has good targeting property, light stability and biological safety, is suitable for early-stage and non-invasive renal function evaluation, and has good clinical application prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a near-infrared II region nanoprobe and its preparation method and application. Background Art

[0002] The kidney is an important metabolic and excretory organ of the human body, and abnormalities in its function often lead to serious systemic diseases. Kidney injury can be caused by various factors such as drug toxicity, ischemia-hypoxia, infection, and high-altitude environment, and is manifested as damage to renal tubular epithelial cells, inflammatory reactions, interstitial fibrosis, and decreased renal function. Due to the deep anatomical position and strong compensatory ability of the kidney, its injury often lacks typical symptoms in the early stage. Therefore, early diagnosis and intervention are of great significance for preventing the progression of the disease to chronic kidney disease and even end-stage kidney disease.

[0003] Traditional renal function assessment mainly relies on biochemical indexes such as serum creatinine and urea nitrogen, as well as urine tests and imaging examinations such as ultrasound and CT. However, these methods have problems such as insufficient sensitivity and specificity, inability to achieve dynamic monitoring, and difficulty in intuitively reflecting changes in renal tissue structure, and it is often difficult to accurately identify them, especially in the early stage of kidney injury.

[0004] In recent years, near-infrared II region (NIR-II, 1000–1700 nm) imaging technology has received extensive attention in the field of biomedicine. Compared with traditional near-infrared imaging (NIR-I, 700–900 nm), NIR-II has deeper tissue penetration ability, higher signal-to-noise ratio, and lower autofluorescence interference, and is particularly suitable for real-time imaging of deep organs in the body (such as the kidney).

[0005] Therefore, there is an urgent need to develop a high-sensitivity and highly specific kidney imaging probe based on NIR-II imaging, which is expected to achieve early non-invasive diagnosis of kidney injury. Summary of the Invention

[0006] The purpose of the present invention is to overcome the problems of response delay, low spatial resolution, and insufficient imaging depth in existing acute kidney injury (AKI) diagnostic methods, and to provide a rapid and non-invasive kidney injury detection strategy based on near-infrared II region (NIR-II) imaging to achieve early and accurate assessment of kidney function.

[0007] To achieve the above purpose, the present invention provides the following technical solutions:

[0008] In the first aspect of the present invention, a near-infrared II region nanoprobe HS-1@PLGA is provided, and the probe is assembled from the following components:

[0009] Poly(lactic-co-glycolic acid);

[0010] Lecithin;

[0011] DSPE-PEG: The degree of polymerization is 2000 - 5000;

[0012] Near-infrared second-region dye HS-1, with the structure

[0013]

[0014] Among them, the mass ratio of the PLGA, lecithin and DSPE-PEG is (3 - 10):(0.5 - 2):(1 - 4), and the concentration of HS-1 is 0.5 - 1 mg / mL.

[0015] Furthermore, the particle size of the near-infrared second-region nanoprobe HS-1 is 90 - 200 nm, and the Zeta potential is -5 to -30 mV.

[0016] Furthermore, the concentration of the PLGA is 1 - 5 mg / mL, the concentration of the lecithin is 0.33 - 1.5 mg / mL, and the concentration of the DSPE-PEG is 1 - 3 mg / mL.

[0017] In the second aspect of the present invention, a preparation method of the near-infrared second-region nanoprobe HS-1@PLGA is provided, and the method includes:

[0018] Dissolve PLGA, lecithin and DSPE-PEG in an organic solvent to form solution A;

[0019] Dissolve HS-1 in deionized water to form solution B;

[0020] Quickly mix solution A and solution B, perform ultrasonic treatment while magnetic stirring to complete the self-assembly of nanoparticles, and obtain the HS-1@PLGA nanoprobe after purification by centrifugation, ultrafiltration and / or dialysis.

[0021] Furthermore, the volume ratio of solution A to solution B is (1 - 3):(1 - 5).

[0022] Furthermore, the organic solvent is selected from one of acetonitrile, ethanol or tetrahydrofuran.

[0023] Furthermore, the conditions of the ultrasonic treatment include: the ultrasonic treatment frequency is 40 - 100 kHz, and the treatment time is 0.1 - 2 hours; the conditions of the magnetic stirring include: the rotation speed is 500 - 1000 rpm, and the time is 5 - 30 hours.

[0024] In the third aspect of the present invention, an application of the near-infrared second-region nanoprobe HS-1@PLGA in the preparation of a renal injury detection product is provided.

[0025] The probe is used for kidney imaging detection. The imaging uses a second near-infrared imaging system with an excitation wavelength matching HS-1 and an imaging band of 1000 - 1500 nm.

[0026] The probe forms a significant fluorescence signal in kidney tissue within 30 minutes after injection, and can distinguish healthy and damaged kidney tissues.

[0027] The second near-infrared nanoprobe HS-1@PLGA is based on a nanocarrier loaded with the second near-infrared fluorescent dye HS-1. After intravenous injection, it can be specifically distributed in kidney tissue in vivo and achieve high-resolution imaging in the second near-infrared band. This imaging signal can clearly present the fluorescence contour of the kidney within 30 minutes after administration, and show differential distribution according to the degree of kidney injury, realizing the rapid identification and dynamic monitoring of the kidney injury status. The HS-1 dye has excellent second near-infrared luminescence properties, with an emission wavelength concentrated between 1000 - 1400 nm, having high photostability and quantum yield; by encapsulating with a highly biocompatible nanomaterial to form a stable probe, it can avoid the rapid metabolism of free dye and background interference, while improving the kidney enrichment efficiency and imaging clarity.

[0028] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0029] The present invention constructs an imaging system with good biodistribution and kidney targeting performance by encapsulating the second near-infrared dye HS-1. This strategy can not only achieve rapid response imaging of kidney injury within 30 minutes after administration, but also has the ability to perform high-resolution and high-contrast imaging on deep organs - especially the kidney, which is significantly superior to traditional NIR-I imaging technology. At the same time, the HS-1 dye used has good biocompatibility and low toxicity, ensuring the safety of the imaging process, providing a reliable technical support for the early, non-invasive, and accurate diagnosis of AKI, and having important clinical significance and application prospects.

[0030] In addition, the kidney injury detection method provided by the present invention has good biosafety. The imaging probe used shows low toxicity, a controllable metabolic pathway, and good tissue tolerance in animals, is suitable for in vivo live imaging experiments of various animal models, and has the potential for clinical translation. Compared with the prior art, the present invention has the following advantages and technical effects:

[0031] (1) Rapid response imaging: The HS-1@PLGA second near-infrared nanoprobe provided by the present invention can achieve significant fluorescence imaging in the kidney region within 30 minutes after tail vein injection, far superior to the delay problem that traditional biomarkers such as serum creatinine take several hours to several days to reflect kidney injury, and significantly improving the early diagnosis efficiency of AKI.

[0032] (2) Strong imaging penetration and high clarity: Based on the imaging window in the NIR-II (1000–1700 nm) band, the present invention has deeper tissue penetration ability and higher signal-to-noise ratio, can clearly image deep organs such as the kidney in vivo, and the imaging quality is superior to traditional means such as near-infrared region I (NIR-I) or ultrasound.

[0033] (3) Excellent biotargeting: The constructed HS-1@PLGA nanoprobe can actively accumulate in the kidney region in vivo, is highly sensitive to the renal function status, can accurately distinguish normal and damaged renal tissues, and is applicable to different types of AKI models (such as cisplatin-induced and ischemia-reperfusion models).

[0034] (4) Good physical and optical stability: Compared with traditional fluorescent probes such as ICG, the probe of the present invention has significantly enhanced stability in the body fluid environment, low fluorescence signal attenuation (>90% fluorescence retention rate), can achieve longer imaging tracking, and is suitable for dynamically observing changes in renal function.

[0035] (5) Good biocompatibility and safety: Animal experiment results show that the probe has a clear metabolic path in vivo, no obvious toxicity, no histological damage to the main organs, and no significant changes in blood routine and biochemical indexes, meeting the requirements for long-term in vivo application.

[0036] (6) Applicable to multiple imaging scenarios: The probe of the present invention shows good applicability and imaging consistency in multiple acute kidney injury models, and has broad potential for experimental research and clinical translation.

[0037] In summary, the present invention is significantly superior to the prior art in terms of response speed, imaging quality, targeting ability, biological safety, etc., provides a new technical path for the early non-invasive diagnosis of acute kidney injury, and has important scientific research value and application prospects. Description of the Drawings

[0038] Figure 1 (a) Absorption spectrum of HS-1@PLGA nanoparticles; (b) Emission spectrum of HS-1@PLGA nanoparticles.

[0039] Figure 2 Particle size of (a) HS-1@PLGA nanoparticles; (b) Zeta potential of HS-1@PLGA nanoparticles.

[0040] Figure 3 Stability comparison of HS-1@PLGA nanoparticles and ICG nanoparticles in PBS and DMEM culture media.

[0041] Figure 4 Stability of HS-1@PLGA nanoparticles in PBS.

[0042] Figure 5 For the imaging effect of HS-1@PLGA nanoparticles on mice with high altitude hypoxia-induced kidney injury.

[0043] Figure 6 For the distribution of HS-1@PLGA nanoparticles in mice.

[0044] Figure 7 For the blood routine and blood biochemical indexes of mice after continuous injection of HS-1@PLGA nanoparticles for 3 days.

[0045] Figure 8 For the HE staining of various organs of mice after continuous injection of HS-1@PLGA nanoparticles for 3 days.

[0046] Figure 9 For the hydrogen spectrum of HS-1. Specific implementation manners

[0047] The following will specifically elaborate on the present invention in combination with specific implementation manners and examples, and the advantages and various effects of the present invention will be presented more clearly therefrom. Those skilled in the art should understand that these specific implementation manners and examples are used to illustrate the present invention, rather than limiting the present invention.

[0048] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as having the meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention pertains. In case of any contradiction, this specification shall prevail.

[0049] The raw material sources of the embodiments of the present invention are as follows:

[0050] PLGA is (50:50) (poly(lactic-co-glycolic acid)(50:50)), a copolymer of polylactic acid (PLA) and polyglycolic acid (PGA), CAS No: 34346-01-5, purchased from MedChemExpress (MCE), catalog number: HY-B2247.

[0051] Lecithin is specifically selected as L-α-phosphatidylcholine, molecular formula: C42H80NO8P, CAS number: 8002-43-5, purchased from Adamas, product number 01155840.

[0052] DSPE-PEG, also known as MPEG2000-DSPE or methoxypolyethylene glycol phospholipid, MW: 2000, purchased from Pengshuo Biotech.

[0053] Unless otherwise specified, all kinds of raw materials, reagents, instruments and equipment used in the present invention can be obtained through market purchase or by existing methods.

[0054] Example 1: Preparation of Near-Infrared II Region Nanoprobe HS-1@PLGA

[0055] I. Synthesis of Compound HS-1

[0056]

[0057] Synthesis of Compound 2: Compound 1 (660 mg, 2.57 mmol) and the solvent tetrahydrofuran (20 mL) were successively added to a round-bottom reaction flask. The reaction flask was placed in a low-temperature reaction kettle, and the temperature was set to -78 °C. After 30 minutes, n-butyllithium (1.24 mL, 3.09 mmol) was added. After 2 hours, tributyltin chloride (0.84 mL, 3.09 mmol) was added. After 12 hours, ethyl acetate was added to the reaction solution to quench the reaction, and it was extracted three times with distilled water to obtain Compound 2. Without purification, it was directly fed for the subsequent reaction.

[0058] Synthesis of Compound 4: Compound 2 (529.5 mg, 0.84 mmol), 2,5-dibromo-3,4-dinitrothiophene (Compound 3, 0.34 mmol, 111.65 mg), the catalyst tetrakis(triphenylphosphine)palladium (0.04 mmol, 38.87 mg), and toluene (80 mL) were successively added to a round-bottom reaction flask. The inside of the reaction flask was under an argon atmosphere, and the temperature was 110 °C. After 18 h, the reaction ended. The reaction temperature was cooled to room temperature, and ethyl acetate was added to terminate the reaction. It was extracted three times with distilled water and once with saturated brine, dried over anhydrous magnesium sulfate, and separated by silica gel column chromatography to obtain Compound 4 (205.93 mg, yield 72%).

[0059] Synthesis of Compound 5: Zinc powder (8.3 g, 69.23 mmol), Compound 4 (970.7 mg, 0.58 mmol), methanol (40 mL), dichloromethane (40 mL), and the inorganic salt ammonium chloride (1.11 g, 20.77 mmol) were added to a round-bottom reaction flask. The reaction was carried out at 25 °C for 2 h. After diluting the reaction solution with dichloromethane, it was extracted three times with saturated sodium chloride. It was dried over anhydrous magnesium sulfate, filtered, and concentrated to obtain an intermediate product. The intermediate product, N-sulfinylaniline (494 mg, 3.56 mmol), trimethylchlorosilane (4.74 mmol, 514 mg), and ultradry pyridine (4 mL) were successively added to the reaction flask. The reaction was carried out at 25 °C for 6 h. After completion, the reaction solution was diluted with dichloromethane, extracted three times with 1 M hydrochloric acid solution, dried over anhydrous magnesium sulfate, concentrated to obtain a crude product, and separated by column chromatography to obtain pure Compound 5 (505.45 mg, yield 52.8%).

[0060] Synthesis of the chemical reactant 14: Compound 5 (1.86 g, 2.19 mmol), N,N-dimethylformamide (10 mL), acetonitrile (5 mL), and N-bromosuccinimide (0.81 g, 4.83 mmol) were successively added into a flask. The reaction was carried out at 65 °C for 12 h. After the temperature of the reaction solution returned to room temperature, the reaction solution was diluted with dichloromethane, extracted three times with saturated sodium bicarbonate solution, and dried over anhydrous magnesium sulfate to obtain Compound 6, which was directly used in the next-step reaction without further purification.

[0061] Synthesis of Compound 8: Compound 6 (1.14 g, 2.1 mmol), the catalyst tetrakis(triphenylphosphine)palladium (97.15 mg, 0.08 mmol), tetrahydrofuran (50 mL), 15% sodium bicarbonate solution (10 mL), and Compound 7 (848.3 mg, 0.84 mmol) were successively added into a reaction flask equipped with a condenser reflux device, and a nitrogen atmosphere was maintained in the reaction flask. The reaction temperature was 80 °C. The reaction was terminated after 18 h. After the temperature of the reaction solution was cooled to room temperature, the reaction solution was diluted with dichloromethane and extracted three times with saturated sodium chloride to obtain a crude product. Compound 7 (777.29 mg, yield 55%) was obtained by column chromatography separation.

[0062] Figure 9 The 1H NMR spectrum of [compound name] is completely consistent with the molecular structure of HS-1, verifying the precise modification of its symmetric conjugated skeleton and hydrophilic substituents.

[0063] II. A near-infrared second-region nanoprobe HS-1@PLGA and its preparation method

[0064] Weigh 2 mg of PLGA (poly(lactic-co-glycolic acid)), 0.4 mg of lecithin, and 0.8 mg of DSPE-PEG2000 and dissolve them in acetonitrile (Solution A).

[0065] Dissolve 675 μg of HS-1 in 1.5 mL of deionized water (Solution B).

[0066] Mix Solution A (500 μL) and Solution B (1.5 mL), perform ultrasonic treatment (50 kHz, 0.5 h), and magnetic stirring (900 rpm, 24 h).

[0067] Purify by ultrafiltration and store in the dark at 4 °C. The particle size is 111.6 nm and the PDI is 0.15.

[0068] Example 2: Renal injury imaging

[0069] 1. Method for detecting renal injury

[0070] Dissolve HS-1@PLGA nanoparticles in sterile PBS buffer to prepare an injection solution. Inject 100 μL of the injection solution into mice via the tail vein. After injection, let it stand for 30 minutes to allow the probe to distribute and accumulate in the kidney region in vivo.

[0071] Use a near-infrared second window (NIR-II) fluorescence imaging system (excitation wavelength matching HS-1) to perform in vivo imaging on mice, with a wavelength band of 1000–1500 nm. The observation results show that the probe produces a significant fluorescence signal in the kidney tissue, and the signal intensity is correlated with the degree of kidney injury.

[0072] In addition, the following experimental verifications were also carried out on HS-1@PLGA nanoparticles:

[0073] Example 3: Determination of absorption and emission spectra

[0074] Use an ultraviolet-visible-near-infrared spectrophotometer and a fluorescence spectrometer to measure their absorption and emission spectra respectively. The results are as Figure 1 shown.

[0075] Figure 1 (a) Absorption spectrum: It shows the absorption peak of HS-1@PLGA in the near-infrared region (e.g., 900 - 1100 nm), indicating its suitability for NIR-II excitation.

[0076] Figure 1 (b) Emission spectrum: It shows that the emission peak is located at 900–1500 nm (main peak is about 1100 nm), covering the NIR-II window and separated from the autofluorescence of background tissues (<900 nm), ensuring a high signal-to-noise ratio.

[0077] Prove the NIR-II optical properties of the probe. Compared with existing NIR-I probes (such as ICG, emission peak is about 800 nm), the emission wavelength is significantly red-shifted, the penetration depth is increased, and the imaging depth is better.

[0078] Example 4: Particle size and Zeta potential of nanoparticles

[0079] Use dynamic light scattering and electrophoretic light scattering techniques for measurement. The results are as Figure 2 shown.

[0080] Figure 2 (a) Particle size distribution: Dynamic light scattering (DLS) shows that the average particle size is about 111.6 nm, and the polydispersity index (PDI) < 0.2, indicating that the particles are uniform.

[0081] Figure 2 (b) Zeta potential: About -3.35 mV, close to neutral, reducing non-specific adsorption with blood components and prolonging the blood circulation time.

[0082] The particle size range (90 - 200 nm) meets the requirements of passive renal targeting (EPR effect), and the Zeta potential (-5 to -30 mV) ensures colloidal stability and avoids aggregation, indicating good colloidal stability.

[0083] Example 5. Stability comparison between HS-1@PLGA and ICG

[0084] Under continuous light irradiation in PBS, the fluorescence intensity retention rates of the two kinds of nanoparticles were compared. The results are as Figure 3 shown.

[0085] As can be Figure 3 seen, the fluorescence retention rate of HS-1@PLGA is higher than 95% within 30 min, and that of ICG nanoparticles is less than 20%, which is significantly better than that of ICG nanoparticles. The probe of the present invention has the advantage of photostability and low fluorescence signal attenuation (>90% retention rate). It proves the protective effect of PLGA encapsulation on the dye and solves the problem of easy quenching of free dye.

[0086] Example 6. Storage stability of HS-1@PLGA in PBS

[0087] Stored in PBS for 3 days, the particle size and fluorescence intensity were detected. The results are as Figure 4 shown.

[0088] As can be Figure 4 seen, the particle size change within 3 days is <5%, the fluorescence intensity retention rate is >90%, and there is no precipitation or aggregation phenomenon. It verifies that the probe has long-term storage stability and meets the requirements of clinical practical applications.

[0089] Example 7. Imaging effect in mice with renal injury induced by high-altitude hypoxia

[0090] The probe was injected into the renal injury model induced by high-altitude hypoxia, and imaging was performed at multiple time points. The probe aggregated in the kidneys within 30 min, with high imaging contrast and could effectively distinguish the pathological state.

[0091] The results are as Figure 5 shown. After 30 min of injecting the probe, NIR-II imaging showed that the signal intensity of the injured kidneys decreased by more than 50% compared with the normal group (for example, the signal of the normal group was 1000 a.u., and that of the injured group was 400 a.u.). It verifies the universality of the probe in a complex model (high-altitude hypoxia) and reflects the technical effect of "being applicable to various renal injuries".

[0092] Example 8. Biodistribution and safety evaluation

[0093] Six hours after injection, the distribution of the nanoparticles in mice is as Figure 6As shown, the probes were mainly enriched in the kidneys (65% of the total), followed by the liver and spleen (about 15% each), and <5% in other organs, indicating that the HS-1@PLGA nanoparticles were mainly enriched in the kidneys.

[0094] Blood routine and blood biochemical indices such as Figure 7 As shown, after 3 consecutive injections, there were no significant differences in indices such as white blood cell count (WBC), liver enzymes (ALT / AST), and creatinine (Cr) compared with the control group (p > 0.05).

[0095] HE staining of major organs was as Figure 8 As shown, sections of organs such as the heart, liver, spleen, lung, and kidney showed normal tissue structures, without inflammation, necrosis, or fibrosis. HE staining after 3 consecutive injections showed no obvious organ damage, and there were no significant abnormalities in blood routine and biochemical indices.

[0096] In summary, the HS-1@PLGA nanosensor constructed in the present invention has good optical properties, biodistribution characteristics, and biosafety, can achieve early, rapid, and non-invasive diagnosis of AKI, and has important scientific research and clinical application values.

[0097] Finally, it should also be noted that the term "comprising", "including" or any other variation thereof is intended to cover non-exclusive inclusion, such that a process, method, article or apparatus comprising a series of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article or apparatus.

[0098] Although the preferred embodiments of the present invention have been described, those skilled in the art can make additional changes and modifications once they learn the basic creative concept. Therefore, the appended claims are intended to be construed to include the preferred embodiments as well as all changes and modifications falling within the scope of the present invention.

[0099] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention is also intended to include these modifications and variations.

Claims

1. A near-infrared second near-infrared region nanoprobe HS-1@PLGA, characterized in that, The probe is assembled from the following components: Poly(lactic-co-glycolic acid); Lecithin; DSPE-PEG: the degree of polymerization is 2000 - 5000; Near-infrared second near-infrared dye HS-1, with the structure Among them, the mass ratio of the PLGA, lecithin, and DSPE-PEG is (3 - 10):(0.5 - 2): (1 - 4), and the concentration of HS-1 is 0.5 - 1 mg / mL.

2. The near-infrared second-region nanoprobe HS-1@PLGA according to claim 1, wherein The particle size of the near-infrared second near-infrared nanoprobe HS-1 is 90 - 200 nm, and the Zeta potential is -5 to -30 mV.

3. The near-infrared second-region nanoprobe HS-1@PLGA according to claim 1, wherein The concentration of the PLGA is 1 - 5 mg / mL, the concentration of lecithin is 0.33 - 1.5 mg / mL, and the concentration of DSPE-PEG is 1 - 3 mg / mL.

4. A method for preparing the near-infrared second-region nanoprobe HS-1@PLGA according to any one of claims 1 to 3, characterized in that, The method includes: Dissolving PLGA, lecithin, and DSPE-PEG in an organic solvent to form solution A; Dissolving HS-1 in deionized water to form solution B; Quickly mixing solution A and solution B, performing ultrasonic treatment while magnetically stirring, completing the self-assembly of nanoparticles, and obtaining the HS-1@PLGA nanoprobe after purification by centrifugation, ultrafiltration, and / or dialysis.

5. The preparation method of the near-infrared second-region nanoprobe HS-1 according to claim 4, characterized in that, The volume ratio of solution A to solution B is (1 - 3):(1 - 5).

6. The preparation method of the near-infrared second-region nanoprobe HS-1 according to claim 4, characterized in that, The organic solvent is selected from one of acetonitrile, ethanol, or tetrahydrofuran.

7. The preparation method of the near-infrared second-region nanoprobe HS-1 according to claim 4, characterized in that, The conditions for the ultrasonic treatment include: the ultrasonic treatment frequency is 40 - 100 kHz, and the treatment time is 0.1 - 2 hours; The conditions for the magnetic stirring include: the rotation speed is 500 - 1000 rpm, and the time is 5 - 30 hours.

8. Use of the near-infrared second near-infrared nanoprobe HS-1@PLGA according to any one of claims 1 - 3 in the preparation of a renal injury detection product.

9. The application according to claim 8, characterized in that, The probe is used for kidney imaging detection, and the imaging uses a near-infrared second near-infrared imaging system, the excitation wavelength matches HS-1, and the imaging band is 1000–1500 nm.

10. The application according to claim 8, wherein The probe forms a significant fluorescence signal in the kidney tissue within 30 minutes after injection, and can distinguish healthy and damaged kidney tissues.

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