Preparation method and application of nano fluorescent compound for targeting tumor cells

Abstract

The invention belongs to the field of medicine, and relates to a preparation method and application of a nano fluorescent compound for targeting tumor cells, the method comprises the following steps: firstly, weighing 20mg of berberine hydrochloride and 30mg of IR-775 chloride, respectively dissolving in 2mL of tetrahydrofuran and 3mL of methanol, mixing, and continuously stirring at 50 DEG C for 30 minutes; and then adding 10mg of distearoyl phosphatidylcholine, continuously stirring at 50 DEG C for 30 minutes, and carrying out rotary evaporation to remove the solvent, thereby finally obtaining the nano fluorescent compound. According to the method, the technical problems that IR-775 and berberine hydrochloride are easy to stack after being compounded, and a luminous compound is not easy to prepare due to an aggregation-induced quenching effect are solved, and tumor targeted imaging is realized.

Description

Technical Field

[0001] This invention belongs to the field of medicine, specifically relating to a method for preparing and applying a nano-fluorescent complex that targets tumor cells. Background Technology

[0002] Precision treatment of malignant tumors has always faced the dual challenges of low drug delivery efficiency and difficulty in monitoring efficacy. With the rapid development of nanotechnology, drug delivery systems have shown great potential in the field of tumor diagnosis and treatment. Nanocarriers can significantly improve drug targeting and bioavailability through precise size control and surface modification. Fluorescent probes, as multifunctional tools, can not only achieve high-resolution cell imaging but also monitor the distribution and release kinetics of drugs in vivo in real time, providing technical support for precision medicine.

[0003] In recent years, nanocomposites with both diagnostic and therapeutic functions have become a research hotspot. They can integrate fluorescence imaging and drug delivery functions to achieve "visualized" tumor treatment, which can significantly improve the accuracy and safety of tumor treatment.

[0004] Berberine hydrochloride (BBR) is an alkaloid extracted from the traditional Chinese medicine Coptis chinensis. It possesses broad-spectrum pharmacological activities, including antibacterial, anti-inflammatory, and antitumor effects. However, its poor water solubility and low bioavailability limit its clinical application. Nanoparticle delivery technology offers a new approach to overcome this limitation; liposomes, through phospholipid bilayer encapsulation, improve the solubility of berberine hydrochloride. However, existing delivery systems are mostly limited to single therapeutic functions, failing to integrate the dual needs of diagnosis and treatment, and lacking real-time monitoring capabilities. Furthermore, existing delivery systems lack pH-responsive characteristics, resulting in poor targeting and difficulty in achieving targeted therapy.

[0005] IR-775 is a near-infrared fluorescent dye with deep tissue penetration and high spatiotemporal resolution, widely used in bioimaging and photodynamic therapy. However, the aggregation-induced quenching effect of IR-775 limits its application at high concentrations, and its complexation with berberine hydrochloride easily results in stacking, producing non-luminescent or even non-luminescent complexes. Therefore, it is necessary to study existing fluorescence imaging and drug delivery systems to design a treatment strategy that enables "visualized" tumor therapy. Summary of the Invention

[0006] In view of the shortcomings and deficiencies of the existing technology, the present invention provides a method for preparing a nano-fluorescent complex targeting tumor cells. The method integrates the anti-tumor activity of berberine hydrochloride (BBR) and the near-infrared fluorescence characteristics of IR-775 chloride into the same delivery system by coating with a distearylphosphatidylcholine (DSPC) structure, thereby achieving tumor-targeted imaging. At the same time, by improving the preparation process, the technical problem of easy stacking of IR-775 and berberine hydrochloride complex, and the difficulty in making luminescent complex due to aggregation-induced quenching effect is solved.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: A method for preparing a fluorescent nanocomposite targeting tumor cells includes the following steps: first, weigh 20 mg of berberine hydrochloride and 30 mg of IR-775 chloride, dissolve them in 2 mL of tetrahydrofuran and 3 mL of methanol respectively, mix them, and stir continuously at 50 °C for 30 min; then add 10 mg of distearylphosphatidylcholine, continue stirring at 50 °C for 30 min, and then remove the solvent by rotary evaporation to finally obtain the fluorescent nanocomposite.

[0008] The fluorescence quantum yield of the nano-fluorescent complex prepared by this invention was 11.12%, 11.78%, and 11.56% in three tests, respectively, indicating that the complex has a certain fluorescence efficiency and can provide sufficient fluorescence signal in the in vivo environment to realize real-time monitoring of drug release and distribution. Furthermore, transmission electron microscopy and atomic force microscopy show that the complex is approximately spherical, relatively uniformly distributed on the substrate, and has a smooth surface. Thus, this invention successfully solves the technical problem that existing IR-775 and berberine hydrochloride complexes are prone to stacking and difficult to form luminescent complexes due to aggregation-induced quenching effects.

[0009] The tumor cell-targeting fluorescent nanocomplex prepared by this invention can enter HCT-116 cells and distribute in the perinuclear region, enabling clearer localization of tumor cells and achieving tumor cell-targeted imaging. Therefore, it can be used in the preparation of reagents for tumor cell imaging and monitoring of tumor treatment effects.

[0010] The fluorescent nanocomplex targeting tumor cells prepared in this invention has a strong inhibitory effect on HCT-116 and A549 cells, and exhibits a concentration-dependent proliferation inhibition effect (maximum inhibition rate of 76.44%), with better anti-tumor effect. At low concentrations, the fluorescent complex has significant cytotoxicity to HCT-116 cells, so it can be used in the preparation of anti-tumor drugs.

[0011] As a preferred embodiment of the present invention, the antitumor drug includes anti-colon cancer drugs and anti-lung cancer drugs.

[0012] As a preferred embodiment of the present invention, the anti-colon cancer drug is a targeted therapy drug.

[0013] Advantages and beneficial effects of the present invention: (1) This invention innovatively constructs an IR-775 chloride-berberine hydrochloride (BBR) fluorescent complex (IBDC) with distearylphosphatidylcholine (DSPC) as the carrier. By encapsulating the structure of DSPC, the antitumor activity of BBR and the near-infrared fluorescence characteristics of IR-775 are integrated into the same delivery system to achieve tumor-targeted imaging. At the same time, by improving the preparation process, the problem of easy stacking of IR-775 and berberine hydrochloride after compounding and the difficulty in making luminescent complex due to aggregation-induced quenching effect is solved.

[0014] (2) The fluorescent complex constructed in this invention is coated with DSPC, which not only solves the problems of poor solubility and low bioavailability of berberine hydrochloride (significantly improves the bioavailability of berberine hydrochloride), but also avoids the problem of IR-775 not emitting light due to aggregation-induced quenching (ACQ) effect, so that the complex has near-infrared fluorescence characteristics and has a high fluorescence quantum yield (11.5%±0.3) in the near-infrared region, which can provide sufficient fluorescence signal in the in vivo environment and realize real-time monitoring of drug release and distribution.

[0015] (3) The fluorescent complex prepared in this invention was characterized by transmission electron microscopy and atomic force microscopy. The complex has a uniform spherical structure and a stable particle size distribution. This spherical structure not only gives the complex a larger specific surface area, which is beneficial to the delivery and release of drugs in vivo, but also gives it high stability and excellent biocompatibility by encapsulating the IR-775 chloride-berberine hydrochloride complex in a phospholipid bilayer.

[0016] (4) The fluorescent complex prepared in this invention was confirmed by gastrointestinal environment simulation release experiment to have pH-responsive drug release characteristics: under simulated tumor microenvironment (pH=6.0) conditions, the cumulative release amount was the highest in 48h, which can locally increase the drug concentration, enhance local efficacy and reduce systemic toxicity; at the same time, the pH=6.0 environment matches the weakly acidic environment of the upper part of the small intestine, which can promote the rapid release of the drug in the jejunum, the main absorption site; the pH=7.0 environment can prolong the sustained release time of the drug in the middle and lower part of the small intestine and improve bioavailability.

[0017] (5) The fluorescent complex prepared in this invention was shown by cell proliferation toxicity experiment (CCK8) to have a strong inhibitory effect on HCT-116 and A549 cells and exhibited a concentration-dependent proliferation inhibition effect (the highest inhibition rate was 76.44%). It had a better anti-tumor effect, and IBDC showed significant cytotoxicity to HCT-116 cells at low concentrations.

[0018] (6) The fluorescent complex prepared in this invention has been confirmed by cell imaging experiments to have good biocompatibility. It can enter HCT-116 cells and be distributed in the perinuclear region, which can more clearly locate tumor cells and achieve tumor cell targeted imaging. At the same time, it allows berberine hydrochloride (BBR) to enter the cell and exert its anti-tumor activity, solving the problem that existing drugs are difficult to reach the target site and difficult to achieve targeted "visual" treatment.

[0019] (7) The fluorescent complex prepared by this invention can efficiently enter tumor cells and distribute precisely, realizing “visualized” treatment and real-time monitoring, improving the accuracy and safety of tumor treatment. This scheme provides a new paradigm for the development of intelligent delivery and integrated diagnosis and treatment technology of natural drugs, and has important scientific significance and clinical application value. Attached Figure Description

[0020] Figure 1 This is a structural diagram of berberine hydrochloride and IR-775 chloride in this invention; Figure 2 This is a schematic diagram of the fluorescent complex (IBDC) and its dimethyl sulfoxide solution of the present invention under natural light and 365nm ultraviolet light; Figure 3 The images show the UV absorption and fluorescence spectra of the fluorescent complex (IBDC) of this invention in different solvents; where a is the UV absorption spectrum and b is the fluorescence spectrum. Figure 4 The fluorescence stability of the fluorescent complex (IBDC) of the present invention in different solvents is shown; where a is 60 seconds and b is 600 seconds. Figure 5 The fluorescence quantum yield of the fluorescent complex (IBDC) of this invention; Figure 6 The proliferation rates of A549 and HCT-116 cells treated with different concentrations of the fluorescent complex (IBDC) are shown; where a is a bar chart and b is a line chart. Figure 7 Transmission electron microscopy image of the IBDC fluorescent complex; Figure 8 AFM image of the IBDC fluorescent complex; Figure 9 Fluorescence microscopy images of HCT-116 cells treated with IBDC; Figure 10 Fluorescence confocal scatter plot of DAPI in the green and red channels; Inset: Pearson correlation coefficient. Figure 11 Photos taken before and after the 1BDC in vitro release test; Figure 12 This represents the cumulative release of IBDC in buffer solutions at different pH values. Detailed Implementation

[0021] 1. Experimental Section: 1.1. Materials and Instruments: All reagents and solvents used in this invention were commercially available products and were not further purified. Absorption spectra were measured using a UV-2550 UV-Vis spectrophotometer; fluorescence emission spectra were measured using a Shimadzu RF-5301PCS fluorescence spectrophotometer; transmission electron microscopy (TEM) characterization and image acquisition were performed using a FEI G2F20 microscope; and zeta potentials were measured using a Zetasizer NanoZS90 nanoparticle size potentiometer. All optical tests were performed at room temperature.

[0022] 1.2. Synthesis of fluorescent complexes: Weigh out 20 mg of berberine hydrochloride and 30 mg of IR-775 chloride (see details for chemical structure). Figure 1 The IBDC nanocomposite was dissolved in 2 mL of tetrahydrofuran (THF) and 3 mL of methanol (MeOH), respectively, and mixed. The mixture was stirred continuously at 50 °C for 30 min. Then, 10 mg of distearylphosphatidylcholine (DSPC) was added, and the mixture was stirred at 50 °C for another 30 min. The solvent was then removed by rotary evaporation, and the dark green IBDC nanofluorescent composite (56 mg, yield: 93%) was finally obtained.

[0023] 1.3. Determination of Spectral Properties: Using a quartz cuvette with an optical path of 1 cm as the container, the UV-Vis spectrum of the fluorescent complex was measured using a UV-2550 UV-Vis spectrophotometer. 2 mg of the fluorescent complex was weighed and dissolved in dichloromethane (DCM), ethanol (EtOH), and tetrahydrofuran (THF), respectively, to prepare appropriate concentration gradients according to experimental requirements. The spectra of solutions at different concentrations were measured and recorded, with the UV detection wavelength range being 400–800 nm. The fluorescence spectrum of the fluorescent complex was measured using 785 nm as the excitation wavelength, with the emission wavelength recorded in the range of 400–900 nm. Furthermore, the fluorescence stability of the fluorescent complex was measured at 60 s and 600 s using dimethyl sulfoxide (DMSO) and ethanol (EtOH), respectively.

[0024] 1.4. Determination of fluorescence quantum yield: The quantum yield of the fluorescent complex was determined using a relative method, with IR125 (fluorescence quantum yield φf = 0.23), a standard substance with similar excitation and emission wavelength ranges, selected as the reference material. Using dimethyl sulfoxide as solvent, absorbance was measured by a UV-Vis spectrophotometer to prepare sample and reference solutions with absorbance intensities ranging from 0.03 to 0.05. Fluorescence spectra were measured using a fluorescence spectrophotometer, and the fluorescence quantum yield was calculated.

[0025] 1.5. In vitro release experiment: Phosphate buffer solutions with different pH values ​​(5.0, 6.0, 7.0) were prepared as release media. 5% sodium dodecyl sulfate (SDS, w / v) was added to each buffer solution. Absorbance was measured at 785 nm, and a sample concentration-absorbance standard curve was plotted. A 2.0 mg / mL drug-loaded micelle solution was prepared, and 2 mL was placed in a dialysis bag (MD34-14000). The dialysis bag was placed in 20 mL of release medium, and release experiments were conducted under isothermal shaking conditions at 37℃ and 130 rpm. Samples of 3 mL were taken at 0.5, 1, 1.5, 2, 3, 4, 5, 6, 8, 10, 12, 16, 20, 24, 36, and 48 h, with 3 mL of fresh buffer added simultaneously. The absorbance of the release medium at 785 nm was measured at each time point. The content of the fluorescent complex in the release medium was calculated based on the standard curve, and the cumulative drug release rate was then calculated.

[0026] 1.6. Transmission Electron Microscopy (TEM) Characterization: Take an appropriate amount of sample and place it in a centrifuge tube. Use ethanol as a solvent and sonicate to make the sample evenly dispersed in ethanol. Use a pipette to take 10 μL of the uniform mixture of sample and ethanol and drop it onto a copper grid. Dry it under an infrared lamp. Finally, place the sample on the sample stage and observe and acquire the sample image using an HT7800 120kV transmission electron microscope.

[0027] 1.7. Atomic Force Microscopy (AFM) Characterization: The sample was uniformly dispersed in ethanol, then coated onto a mica sheet and allowed to air dry. Finally, the mica sheet was placed on the atomic force microscope (AFM) stage for scanning characterization.

[0028] 1.8. Cell proliferation toxicity assay (CCK8): Cell pretreatment: HCT-116 cells (purchased from Wuhan Pronosai Life Science Technology Co., Ltd.) and A549 cells (purchased from Xiamen Yimo Biotechnology Co., Ltd.) in logarithmic growth phase were digested with trypsin and prepared into a cell suspension of appropriate concentration. The cells were seeded at a density of 5000 cells / well in 96-well plates, with 8 complex wells and 2 control wells in each group.

[0029] Cell drug treatment: According to the experimental group, IBDC solution of blank control group (no sample), 20, 40, 60, 80, 100, and 120 μg / mL was added to the cells in each well, and the cells were co-cultured for 24 h.

[0030] CCK8 colorimetric reaction: After culturing for 24 hours, remove the 96-well plate, add 10 μL of CCK8 solution to each well, and incubate in an incubator for 1 hour; measure the absorbance at 450 nm using a microplate reader.

[0031] 1.9. Cell imaging experiment: Cell pretreatment: HCT-116 cells in logarithmic growth phase were seeded onto cell crawling plate sections in 12-well plates, with 5 × 10⁶ cells per plate. 4 One cell was cultured for another 24 hours.

[0032] Cell drug treatment: The IBDC fluorescent complex was prepared into a stock solution of 0.5 mg / mL with dimethyl sulfoxide; after the cells adhered and grew for 24 h, the sample was diluted with culture medium to a final concentration of 5 μg / mL; the original culture medium was discarded, and after washing with phosphate-buffered saline (PBS), 1 mL of the diluted sample was added to each well, and the cells were cultured for another 24 h.

[0033] Microscopic observation: After 24 hours of drug treatment, add an appropriate amount of Hoechst staining solution (final concentration 5 μg / mL), discard the cell culture medium in each well, and add 500 μL of diluted staining solution; after incubation at 37℃ for 10-15 minutes, observe the staining effect under a fluorescence microscope, and take pictures at an appropriate wavelength using a laser confocal microscope.

[0034] 2. Results and Discussion: 2.1 Synthesis Results: This invention employs a one-pot method to prepare fluorescent complexes. The design is as follows: First, berberine hydrochloride and IR-775 chloride combine via electrostatic interaction to form a complex. Then, DSPC phospholipids are used to encapsulate this complex, forming a nano-fluorescent complex. DSPC not only improves the stability and drug delivery performance of the complex system but also enhances its biocompatibility. The fluorescent complex was observed under 365nm ultraviolet light and natural light, and the results are as follows. Figure 2 As shown, the IBDC fluorescent complex and its dimethyl sulfoxide solution both appear dark green under natural light and pale green under 365 nm ultraviolet light.

[0035] 2.2 Spectral properties: The UV-Vis spectroscopic properties of the IBDC fluorescent complex were investigated using dichloromethane (DCM), ethanol (EtOH), and tetrahydrofuran (THF) as solvents, respectively. Figure 3As shown in Table 1, at the same concentration, the IBDC fluorescent complex exhibits the highest absorbance in dichloromethane and the lowest absorbance in ethanol. The maximum absorption wavelengths of the IBDC fluorescent complex in the UV absorption spectrum are 778 nm, 785 nm, and 786 nm in the three solvents, respectively. Furthermore, the absorbance of the IBDC fluorescent complex increases significantly with increasing concentration in all three solvents. The absorption peaks of IR-775 dyes are mostly concentrated in the 770–790 nm range, which matches the 785 nm excitation wavelength selected in this invention, indicating that this invention successfully preserves the characteristic absorption properties of IR-775. Figure 3 As shown in b and Table 1, in all three solvents, the fluorescence intensity of the fluorescent complex decreased with increasing concentration, and the fluorescence peak shifted towards longer wavelengths with increasing concentration, exhibiting a slight redshift. Although the fluorescence intensity decreased, its near-infrared emission characteristics (excitation wavelength 785 nm, emission wavelength 800~900 nm) gave it excellent deep tissue penetration capability for tumor imaging. Figure 4 The results showed that when dimethyl sulfoxide and ethanol were used as solvents, the fluorescence intensity of the IBDC fluorescent complex was almost unaffected at 60s and 600s, respectively, which met the experimental requirements for cell imaging.

[0036] Table 1. UV absorption and fluorescence spectra of IBDC fluorescent complexes in different solvents

[0037] 2.3. Fluorescence quantum yield: Fluorescence quantum yield (φf) is an important parameter for measuring the energy utilization efficiency of fluorescent materials, representing the proportion of fluorescent materials that emit fluorescent photons after absorbing photons. This invention uses IR125 as a standard (φf = 0.23 in dimethyl sulfoxide) to determine and calculate the fluorescence quantum yield of the IBDC fluorescent complex. Figure 5 As shown, the results of the three fluorescence quantum yield tests were 11.12%, 11.78%, and 11.56%, respectively, indicating that the fluorescent complex has a certain fluorescence efficiency and can provide sufficient fluorescence signal in the in vivo environment to realize real-time monitoring of drug release and distribution.

[0038] 2.4. Cell proliferation toxicity assay (CCK8): To evaluate the cytotoxicity of the IBDC fluorescent complex, HCT (HCT-116) and A549 tumor cells were selected for CCK8 assay. Figure 6As shown in Table 2, IBDC inhibited the proliferation of both cell types, and the inhibitory effect on both tumor cell types increased with increasing concentration of the fluorescent complex. Table 2 shows that at a concentration of 120 μg / mL, the survival rates of A549 cells and HCT cells treated with IBDC were 29.43% and 23.56%, respectively. Even at a low concentration of 20 μg / mL, IBDC exhibited strong antitumor activity against HCT cells, significantly higher than its inhibitory effect on A549 cells, indicating that IBDC at low concentrations has significant cytotoxicity against HCT cells.

[0039] Table 2. Proliferation rate of A549 and HCT cells treated with different concentrations of IBDC

[0040] 2.5. Transmission Electron Microscopy (TEM) Characterization: To more intuitively observe the microscopic morphological characteristics of the fluorescent complex, this invention employs transmission electron microscopy to observe the internal morphology of the IBDC fluorescent complex. For example... Figure 7 As shown, IR-775 and berberine hydrochloride are encapsulated by DSPC to form a spherical structure. This spherical structure not only gives the complex a larger specific surface area, which is beneficial for drug delivery and release, but also endows it with high stability and excellent biocompatibility (mimicking cell membrane structure and reducing immunogenicity) by encapsulating the dye-drug complex in a phospholipid bilayer.

[0041] 2.6. Atomic Force Microscopy (AFM) Characterization: To evaluate the stability and large-area uniformity of the IBDC fluorescent complex, the samples were imaged using atomic force microscopy. Figure 8 The image shows the atomic force microscopy (AFM) morphology of the fluorescent complex. The results indicate that the complex particles are approximately spherical, relatively uniformly distributed on the substrate, and have a smooth surface. This result indirectly verifies the spherical structure of the complex observed by transmission electron microscopy (TEM), consistent with the TEM characterization results.

[0042] 2.7. Cell imaging experiments: To clarify the ability of drugs to penetrate tumor cells and their intracellular distribution characteristics, this invention uses HCT-116 cells as a model to conduct fluorescence imaging studies of the IBDC fluorescent complex. A comprehensive imaging analysis was performed using multiple methods, including bright-field imaging, DAPI staining, green channel imaging, red channel imaging, and combined imaging. Figure 9 As shown, the fluorescent complex exhibits strong and clear fluorescence signals in both the green and red channels; the combined imaging results show that IBDC can penetrate the tumor cell membrane and is mainly located around the cell nucleus.

[0043] To further evaluate the interaction between the fluorescent complex and cells, this invention uses ImageJ software to analyze the colocalization of green channel, red channel, and DAPI staining, calculates the Pearson correlation coefficient, and plots a fluorescence confocal scatter plot. Figure 10 The results showed that the Pearson correlation coefficients of colocalization analysis of multiple HCT-116 cells were all greater than 0, ranging from 0.0707 to 0.2849, indicating that DAPI staining was positively correlated with both red and green channels, suggesting that the fluorescent complex can effectively enter the cell and reach the nuclear region, distributing around the cell nucleus.

[0044] 2.8. In vitro release experiment: To investigate the release of IR-775 chloride-berberine hydrochloride after DSPC encapsulation in the gastrointestinal tract, this invention analyzed the in vitro release behavior of the fluorescent complex in three different pH environments. Figure 11 This image shows the physical phenomena of the three stages of in vitro release of the IBDC fluorescent complex. This invention determined the cumulative percentage of drug release from the IBDC fluorescent complex over time, and the results are as follows... Figure 12 As shown, in an environment with pH=6.0, drug release generally exhibits three phases: 0–12 h is the initial rapid release phase, 12–36 h is the middle slow release phase, and 36–48 h is the late gradual release phase. Overall, the cumulative drug release is highest at pH=6.0, followed by pH=7.0, and lowest at pH=5.0. This phenomenon may be due to the partial protonation of the phosphate groups (dissociation constant pKa≈6.5) of DSPC at pH=6.0, leading to a looser phospholipid bilayer and opening of drug diffusion channels. Simultaneously, the pH=6.0 environment matches the weakly acidic environment of the upper small intestine, promoting rapid drug release in the jejunum, the primary absorption site. The pH=7.0 environment prolongs the sustained-release time of the drug in the middle and lower small intestine, improving bioavailability. Furthermore, the high drug release rate at pH=6.0 is compatible with the tumor microenvironment (pH≈6.5–6.8), which can locally increase drug concentration, enhance efficacy, and reduce systemic toxicity. The results of the in vitro release experiment were consistent with the high killing effect of the drug on HCT-116 cells in the cytotoxicity experiment.

[0045] The above-described specific examples are for illustrative purposes only and are not intended to limit the scope of the invention. Those skilled in the art can make various simple deductions, modifications, or substitutions based on the principles of this invention. Therefore, the scope of protection of this invention should be determined by the scope of the claims.

Claims

1. A method of preparing a nano-fluorescent complex targeting tumor cells, characterized in that, The method includes the following steps: First, weigh 20 mg of berberine hydrochloride and 30 mg of IR-775 chloride, dissolve them in 2 mL of tetrahydrofuran and 3 mL of methanol respectively, mix them, and stir continuously at 50 °C for 30 min; then add 10 mg of distearylphosphatidylcholine, continue stirring at 50 °C for 30 min, and then remove the solvent by rotary evaporation to finally obtain the nano-fluorescent composite.

2. The nano-fluorescent complex targeting tumor cells prepared according to claim 1 is used in the preparation of reagents for tumor cell imaging and monitoring of tumor treatment efficacy.

3. The application of the tumor cell-targeting nanofluorescent complex prepared according to claim 1 in the preparation of antitumor drugs.

4. The application according to claim 3, characterized in that, The anti-tumor drugs include anti-colon cancer drugs and anti-lung cancer drugs.

5. The application according to claim 4, characterized in that, The aforementioned anti-colon cancer drug is a targeted therapy drug.

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