Complex β-CD-BDP⊃CPT-ss-PEG and its application

Abstract

The application designs and synthesizes an anticancer photochemotherapy drug, a novel infrared / near-infrared BODIPY photosensitizer BDP, and then constructs an amphiphilic supramolecular nanodrug based on a molecular recognition reaction between beta-cyclodextrin (beta-CD) and a chemotherapy drug CPT. In the supramolecular nanodrug system, the photosensitizer BDP is covalently connected to the macrocyclic host beta-CD. Under the irradiation of a laser, BDP can convert oxygen around tumor tissue into singlet oxygen, so that photodynamic therapy of the tumor can be realized. In the chemotherapy guest, a thio ketone bond sensitive to singlet oxygen is introduced, so that the chemotherapy of the tumor can be realized at the same time.

Description

Technical Field

[0001] The present invention belongs to the technical field of organic synthesis route design and preparation of raw materials and intermediates, and particularly relates to an anticancer photochemotherapeutic drug. The intermediates, their preparation methods and applications in tumor treatment. Background Art

[0002] Chemotherapy is a key treatment option for cancer. However, traditional chemotherapy often faces a series of challenges, such as poor solubility and stability of chemotherapeutic drugs under physiological conditions, the development of drug resistance, and severe toxic side effects on normal tissues, which severely hinder its clinical application. Furthermore, most anticancer drugs, such as paclitaxel, gemcitabine, and cisplatin, exhibit only weak or no fluorescence, making their metabolism in the body difficult to monitor, hindering medical research into the mechanisms of cancer treatment.

[0003] (1) Due to the poor solubility of the anticancer drug CPT and its tendency to transform from a lactone structure to a carboxylate structure in a physiological environment, CPT has not been applied to the clinical treatment of cancer. To solve this problem, we introduced a supramolecular strategy. In the construction of amphiphilic supramolecular nanomedicines, CPT molecules are first covalently linked to a chemotherapy guest with singlet oxygen responsiveness, and then through the host-guest reaction between β-CD and CPT, CPT is complexed into the large hydrophobic cavity of cyclodextrin. Therefore, the construction of supramolecular nanomedicines can not only greatly increase the solubility of CPT chemotherapy drugs, but also maintain its lactone structure with anticancer activity, thus providing a new idea for the application of CPT in clinical treatment.

[0004] (2) Although chemotherapy is an important treatment method in cancer treatment, its clinical application has been severely limited due to its nonspecificity and strong systemic toxicity. To solve this problem, we used a supramolecular strategy to combine photodynamic therapy and chemotherapy for cancer. The near-infrared absorbing photosensitizer BDP was connected to the macrocyclic host β-CD. Under laser irradiation, BDP can convert oxygen in the tumor tissue into singlet oxygen with lethal effect. 1 O2, thereby enabling photodynamic therapy of tumors. Singlet oxygen accumulated at the tumor site simultaneously cleaves the thioketal bond in the chemotherapeutic agent, subsequently inducing a cascade reaction of CPT release, thereby enabling in situ chemotherapy of the tumor. The synergistic treatment of these two approaches based on supramolecular chemistry not only effectively inhibits the growth of superficial tumor tissue but also opens a new window for the treatment of deeper tumor tissue. Summary of the Invention

[0005] In order to solve the defects of poor solubility of CPT and strong toxicity of chemotherapy system in cancer treatment in the prior art, the present invention provides an anticancer photochemotherapeutic drug. A new method for preparing and synthesizing its intermediates.

[0006] The purpose of the present invention can be achieved through the following technical solutions:

[0007] In a first aspect, the present invention provides a complex The complex It is formed by complexing the compound represented by formula A and the compound represented by formula B;

[0008]

[0009] In formula B, n=100-10000 (preferably 2000).

[0010] In a second aspect, the present invention provides a complex The preparation method is as follows:

[0011] The compound of formula A (β-CD-BDP) and the compound of formula B (CPT-ss-PEG) are added to a solvent and dispersed evenly (complexation occurs). The resulting mixture is post-treated H to obtain the complex The molar ratio of the compound of formula A to the compound of formula B is 1:1;

[0012]

[0013] In formula B, n=100-10000 (preferably 2000).

[0014]

[0015] Furthermore, the solvent is dimethyl sulfoxide, tetrahydrofuran, dimethylformamide or water (any solvent that can dissolve compounds A and B, preferably water).

[0016] Furthermore, the volume of the solvent is 5-50 mL / g based on the total mass of the compound of formula A and the compound of formula B.

[0017] Further, the post-treatment H is: freeze-drying the mixed solution to obtain the complex

[0018] Furthermore, the present invention also provides a method for preparing a compound of formula A, wherein the compound of formula A is synthesized as follows:

[0019]

[0020] (1) Preparation of OTs-CD: β-cyclodextrin was added to water to obtain a (turbid) cyclodextrin solution, and an aqueous NaOH solution was slowly added dropwise (the solution became light yellow and clear after the addition), and an acetonitrile solution of toluenesulfonyl chloride was added dropwise (a precipitate was immediately generated). After the addition, the resulting mixed solution was stirred at room temperature for 2-5 hours. The filtrate was collected and the pH was adjusted to 7-8 with a saturated ammonium chloride solution (a precipitate was precipitated). The precipitate was collected by centrifugation and subjected to post-treatment A to obtain compound 1 (OTs-CD).

[0021] The molar ratio of the β-cyclodextrin, the NaOH contained in the NaOH aqueous solution, and the methylbenzenesulfonyl chloride contained in the acetonitrile solution of methylbenzenesulfonyl chloride is 1:1-10:1-5 (preferably 1:3.4:1.1);

[0022]

[0023] (2) Preparation of NH2-CD: Compound 1 (OTs-CD) described in step (1) is reacted with ethylenediamine at 50-90°C for 2-8 hours (preferably at 75°C for 4 hours), and the resulting reaction solution B is subjected to post-treatment B to obtain compound 2;

[0024] The volume of the ethylenediamine is 2-10 mL / g (preferably 5 mL / g) based on the mass of the compound 1.

[0025]

[0026] (3) Preparation of BDP-NHS: Compound 3 (BDP-COOH) was dissolved in dimethylformamide A (DMF), followed by the addition of N-hydroxysuccinimide (NHS), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), and 4-dimethylaminopyridine (DMAP). The mixture was reacted at room temperature for 12-36 h (preferably 24 h). After the reaction, the solvent was evaporated to dryness, and the crude product was washed with methanol (twice) to obtain compound 4 (BDP-NHS);

[0027] The molar ratio of the compound 3, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 4-dimethylaminopyridine is 1:1-5:1-5:1-3, preferably 1:2:3:1.6.

[0028]

[0029]

[0030] (4) Preparation of β-CD-BDP: Dissolve the compound 4 (BDP-NHS) described in step (3) in dimethylformamide B (DMF), add the compound 2 (NH2-CD) described in step (2) and triethylamine, and react at room temperature for 12-36 hours. The resulting mixed solution is subjected to post-treatment D to obtain the compound of formula A (purple solid);

[0031]

[0032] The molar ratio of compound 4, compound 2 and triethylamine is 1:0.2-1:0.1-0.5 (preferably 1:0.625:0.25).

[0033] Furthermore, the volume of the water in step (1) is 5-10 mL / mmol (preferably 8.6 mL / mmol) based on the amount of the β-cyclodextrin.

[0034] Furthermore, the NaOH aqueous solution in step (1) is obtained by dissolving NaOH in water A, and the volume of water A is 0.1-0.2 mL / mmol (preferably 0.18 mL / mmol) based on the amount of NaOH substance.

[0035] Furthermore, the acetonitrile solution of methylbenzenesulfonyl chloride in step (1) is obtained by dissolving methylbenzenesulfonyl chloride in acetonitrile, and the volume of acetonitrile is 0.1-0.3 mL / mmol (preferably 0.284 mL / mmol) based on the amount of methylbenzenesulfonyl chloride.

[0036] Furthermore, the post-treatment A in step (1) is: washing the precipitate with water (once) and acetone (twice), and vacuum drying to obtain the compound shown in formula 1.

[0037] Furthermore, the post-treatment B in step (2) is: cooling the reaction solution B to room temperature, adding acetone A (the volume of the acetone A is 10-100 mL / g, preferably 20 mL / g, based on the mass of the compound 1) for precipitation, centrifuging, dissolving the resulting precipitate in water, and precipitating again with acetone B (the volume of the acetone B is 20-100 mL / g, preferably 20 mL / g, based on the mass of the compound 1), centrifuging, and vacuum drying the resulting precipitate to obtain compound 2.

[0038] Furthermore, the volume of dimethylformamide A in step (3) is 50-200 mL / g (preferably 100 mL / g) based on the mass of compound 3.

[0039] Furthermore, the volume of dimethylformamide B in step (4) is 100-1000 mL / g, preferably 125 mL / g, based on the mass of compound 4.

[0040] Furthermore, the post-treatment D in step (4) is: adding the mixed solution dropwise into acetone C for precipitation, centrifuging, washing the obtained precipitate with acetone (2-4 times), and vacuum drying to obtain the compound of formula A.

[0041] Furthermore, the volume of acetone C in step (4) is 0.2-2 mL / mg (preferably 1 mL / mg) based on the mass of compound 4.

[0042] Furthermore, the compound of formula B is prepared as follows:

[0043] Synthesis of CPT-ss-PEG prodrug

[0044]

[0045] Compound 5 (mPEG n -COOH) is dissolved in dichloromethane (DCM), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 4-dimethylaminopyridine (DMAP) and compound 6 (CPT-ss-OH) are added, and the reaction is carried out at room temperature for 8-16 hours. The resulting reaction solution F is subjected to post-treatment F to obtain the compound of formula B;

[0046]

[0047] The molar ratio of the compound 5, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 4-dimethylaminopyridine and compound 6 is 1:1-4:0.5-2:0.5-2 (preferably 1:2.4:1.2:1.2).

[0048] In Formula 5 and Formula B, n=100-10000 (preferably 2000).

[0049] Furthermore, the volume of dichloromethane is 10-40 mL / g, preferably 28.6 mL / g, based on the mass of compound 5.

[0050] Furthermore, the post-treatment F is: adding water to the reaction solution F for extraction, taking the aqueous phase and adding dichloromethane for extraction, combining the organic phases, and drying to obtain the compound of formula B.

[0051] In a third aspect, the present invention further provides a use of the above complex in the preparation of anti-tumor phototherapy drugs.

[0052] Preferably, the tumor is a 4T1 tumor.

[0053] Acetone A, B, and C are simply for the purpose of distinguishing acetone added at different stages for ease of description and have no other special meaning. Dimethylformamide A and B are simply for the purpose of distinguishing dimethylformamide added at different stages for ease of description and have no other special meaning. Water A is also for the purpose of convenience and has no special meaning.

[0054] Compared with the prior art, the present invention has the following advantages: the present invention designs and synthesizes a novel infrared / near-infrared BODIPY photosensitizer BDP, and then constructs an amphiphilic supramolecular nanodrug based on the molecular recognition reaction between β-cyclodextrin (β-CD) and the chemotherapy drug CPT. In this supramolecular nanodrug system, the photosensitizer BDP is covalently linked to the macrocyclic β-CD host. Under laser irradiation, BDP converts oxygen surrounding tumor tissue into singlet oxygen, thereby enabling photodynamic therapy of tumors. Simultaneously, CPT is covalently linked to a prodrug with a thioketal bond. Under the induction of singlet oxygen, the thioketal bond cleaves, allowing for targeted drug release. This supramolecular nanodrug construction not only significantly improves the solubility of the anticancer drug CPT but also preserves its active structure (lactone), thereby enhancing CPT's chemotherapeutic efficacy. Furthermore, the PEG shell of the supramolecular nanodrug prevents protein absorption, while multiple hydrogen bonds between the cyclodextrin core and π-π stacking interactions between CPT stabilize the supramolecular nanodrug, effectively extending its metabolic cycle time and reducing its systemic toxicity. In vitro cellular and in vivo mouse studies demonstrated that the constructed supramolecular nanodrug exhibits minimal systemic toxicity, good biocompatibility, and effective tumor inhibition. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 Intermediate chemical compound OTs-CD 1 H NMR spectra

[0056] Figure 2 It is the intermediate compound β-CD-BDP 1 H NMR spectra

[0057] Figure 3 It is the intermediate compound CPT-ss-PEG 1 H NMR spectra

[0058] Figure 4 is β-CD-BDP, CPT-ss-PEG and of 1 H NMR spectra

[0059] Figure 5 yes TEM image of nanomedicine

[0060] Figure 6 yes DLS images of nanomedicine

[0061] Figure 7 Changes in the UV absorption curve of DPBF in the presence of BDP-CD in DMSO and H2O when irradiated with 660 nm laser.

[0062] Figure 8 yes Photothermal properties of nanomedicine

[0063] Figure 9 (a) Cell viability of 4T1 cells under light and dark conditions after incubation with different concentrations of β-CD-BDP for 24 h (660 nm, 0.8 W cm -2 ); (b) Cell viability of 4T1 cells under light and dark conditions after incubation with different concentrations of β-CD-BDP for 24 h (660 nm, 0.5 W cm -2 ); (c) Cell viability of 4T1 cells incubated with different concentrations of CPT and CPT-ss-PEG for 24 h; (d) Different concentrations of Cell viability (660 nm, 0.5 W cm-2) under light and dark conditions after incubation of 4T1 cells for 24 h -2 )

[0064] Figure 10 (a) Body weight changes of mice in different treatment groups; (b) Photos of tumors in different treatment groups; (c) Changes in tumor volume of mice in different treatment groups; (d) Weights of important organs and tumors of mice in different treatment groups. DETAILED DESCRIPTION

[0065] For ease of understanding, the present invention will be described in detail below through specific embodiments. It should be noted that the specific embodiments are for illustration only, and it is obvious that those skilled in the art can make various modifications to the present invention within the scope of the present invention based on the description herein.

[0066] Example 1: Preparation of intermediate OTs-CD

[0067] First, 40 g (32.6 mmol) of β-CD was added to 280 mL of water to produce a turbid cyclodextrin solution. Next, 4.32 g (109.4 mmol) of NaOH was dissolved in 20 mL of water and slowly added dropwise to the cyclodextrin solution. After addition, the solution became clear and pale yellow. 6.72 g (35.2 mmol) of p-toluenesulfonyl chloride was dissolved in 10 mL of acetonitrile and slowly added to the mixed solution. The dropwise addition of the toluenesulfonyl chloride solution immediately produced a precipitate. After the addition was complete, the mixed solution was stirred at room temperature for 3 hours. The filtrate was collected and saturated ammonium chloride solution was continuously added until a precipitate formed. The pH of the solution was adjusted to 7-8. The precipitate was collected by centrifugation. The precipitate was washed once with water and twice with acetone before being vacuum dried to yield 4.2 g of a solid product in a 10% yield. Finally, nuclear magnetic resonance (NMR) characterization was performed.

[0068] Figure 1 It is the intermediate chemical compound OTs-CD 1 The H NMR spectrum confirmed the successful preparation of the compound.

[0069] Example 2: Preparation of intermediate NH2-CD

[0070] 2.0 g of OTs-CD was reacted with an excess of EDA (10 mL) at 75°C for 4 h. After the reaction, the mixture was cooled to room temperature and precipitated by the addition of 40 mL of acetone. The precipitate was centrifuged (8000 rpm for 3 min) and dissolved in 5 mL of water. The precipitate was again precipitated with acetone and centrifuged (8000 rpm for 3 min). The precipitate was dried under vacuum at 50°C to obtain 1.8 g of the product (90% yield).

[0071] Example 3: Synthesis of intermediate BDP-NHS

[0072] 50 mg (0.05 mmol) of BDP-COOH (BDP-COOH reference Triggered In Situ Dimorphic Transformation of BF2-Azadipyrromethene Nanoaggregates for Enhanced Solid Tumor Penetration. ACS Nano. 2020 Mar 24; 14 (3): 3640-3650) was dissolved in 5 mL of DMF, followed by the addition of 11.5 mg (0.1 mmol) of NHS (N-hydroxysuccinimide), 29.6 mg (0.15 mmol) of EDCI (1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride) and 10 mg of DMAP (4-dimethylaminopyridine, 0.08 mmol). The reaction was carried out at room temperature for 24 h. After the reaction, the solvent was evaporated to dryness, washed once with methanol, and dried to obtain 40 mg of the product as a purple solid with a yield of 80%.

[0073] Example 4: Synthesis of intermediate β-CD-BDP

[0074] First, 40 mg (0.04 mmol) of BDP-NHS produced in the previous step was dissolved in 5 mL of DMF. Then, 30 mg (0.025 mmol) of NH2-β-CD and 1.5 μL (0.01 mmol) of triethylamine were added and allowed to react at room temperature for 24 hours. The resulting mixture was added dropwise to 40 mL of acetone to obtain a precipitate. The precipitate was washed twice with acetone and dried under vacuum to yield 80 mg of a purple solid (88% yield).

[0075] Figure 2 Intermediate chemical compound β-CD-BDP 1 The H NMR spectrum confirmed the successful preparation of the compound.

[0076] Example 5: Synthesis of CPT-ss-PEG prodrug

[0077] First, 350 mg (0.17 mmol) of mPEG 2000-COOH (methoxy-polyethylene glycol-carboxyl, average molecular weight of 2000) was dissolved in 10mL DCM, followed by the addition of 77.6mg (0.4mmol) EDCI, 24.5mg (0.2mmol) DMAP, and 111mg (0.2mmol) CPT-ss-OH. (CPT-ss-OH reference Dynamic core crosslinked camptothecin prodrug micelles with reduction sensitivity and boronic acid-mediated enhanced endocytosis: An tumor-targeted delivery nanoplatform, International Journal of Pharmaceutics, Volume 580, 2020, 119250.) The reaction was allowed to proceed at room temperature for 12h. After the reaction, the reaction solution was extracted with 20mL HO, and the aqueous phase was extracted again with 20mL dichloromethane. The organic phases were combined and evaporated to dryness to obtain 340mg of the product as a light yellow solid with a yield of 76.4%.

[0078] Figure 3 It is a CPT-ss-PEG prodrug 1 The H NMR spectrum confirmed the successful preparation of the compound.

[0079] Example 6: Synthesis of nanomedicines

[0080] First, 30 mg (0.01 mmol) of β-CD-BDP and 26 mg (0.01 mmol) of CPT-ss-PEG were dissolved in 5 ml of deionized water, then ultrasonicated in an ice bath for 1 h, and finally freeze-dried for 48 h to obtain Pharmaceutical powder.

[0081] Example 7: Study on the complexation of β-CD-BDP and CPT-ss-PEG

[0082] In order to study the host-guest interaction between β-CD-BDP and CPT-ss-PEG, β-CD-BDP, CPT-ss-PEG and carried out 1 H NMR spectroscopy analysis, such as Figure 4As shown in the figure, when 15 mg (0.005 mmol) of β-CD-BDP and 13 mg (0.005 mmol) of CPT-ss-PEG in a 1:1 molar ratio were complexed in 1 mL of deuterated DMSO, the chemical environment of the protons on CPT was observed to change significantly. The double peak of protons at 5.53 ppm became a single peak, which may be caused by the electrostatic interaction between the macrocyclic unit and CPT. In addition, β-CD-BDP and CPT-ss-PEG generated a new proton peak at 2.1 ppm. All these results provide strong evidence for the host-guest complexation between β-CD-BDP and CPT-ss-PEG.

[0083] Example 8: Size characterization of nanomedicines

[0084] Weigh 5 mg of the The drug powder was dissolved in 10 mL of deionized water. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) were then performed to characterize the size of the material. Figure 5 As shown, It can self-assemble into nanoparticles with a particle size of about 100nm in aqueous solution. Figure 6 As shown, DLS also measured The particle size is about 200 nm, which is consistent with the TEM structure. Therefore, it can be proved that β-CD-BDP can undergo host-guest self-assembly with CPT-ss-PEG to form nanoparticles in the range of 100 nm-200 nm. Nanoparticles of this size can accumulate at the tumor site through the EPR effect (hypertonic retention effect), thereby improving the therapeutic efficiency of nanomedicine.

[0085] Example 9: β-CD-BDP 1 Study on O2 Quantum Yield

[0086] 1,3-Diphenylisobenzofuran (DPBF) reacts with singlet oxygen to form endoperoxides, which results in a decrease in its maximum absorption peak at 417 nm. Therefore, the DPBF titration method can be used to study the photosensitizer 1 O2 quantum yield. First, prepare a 1 mg / mL DPBF DMSO solution, then aspirate 30 μL of it and add it to a 20 μM β-CD-BDP aqueous solution or DMSO, shake it evenly, and use a 660 nm laser (0.5 W cm -2 ) for irradiation. Figure 7As shown in a and b, after only 10 seconds, the absorption peak of DPBF at 417 nm decreased significantly, indicating that β-CD-BDP has a strong ability to generate ROS in water or organic solvents, thus having an efficient photodynamic therapy (PDT) effect. 1 O2 is generated by β-CD-BDP, and the -ss- bond in CPT-ss-PEG will consume 1 O2, so the experiment only studied β-CD-BDP.

[0087] Example 8: Study on the photothermal properties of

[0088] To evaluate The photothermal effect of nanoparticles was studied by dissolving 5.6 mg of the nanodrug in Example 6 in 2 mL of water to prepare a 500 μM stock solution, which was then diluted to concentrations of 150, 100, 50, and 25 μM. 200 μL of each aqueous solution was added to a 96-well plate, irradiated from the bottom up with a laser, and the temperature was detected with an infrared temperature camera. -2 ) irradiation aqueous solution for 300s and monitor their temperature rise curves. Figure 8 As shown in a, the temperature of the water in the control group hardly changed after irradiation, while The temperature of the aqueous solution increases significantly, and the heating capacity is The concentration of aqueous solution ( Figure 8 a) and light power ( Figure 8 b) is positively correlated, indicating It can effectively convert light energy into heat energy. Figure 8 As shown in c, After 4 heating and cooling cycles, the maximum temperature did not drop significantly, indicating that Has good photochemical stability. Figure 8 As shown in d, through the light-to-heat conversion formula, The photothermal conversion efficiency can reach 45%.

[0089] Example 10: Cell viability assay

[0090] 4T1 (mouse breast cancer) cells were inoculated into 96-well culture plates and cultured in a 5% CO2, 37°C incubator for 24 h. Then, β-CD-BDP (with or without 0.8 or 0.5 W cm-2 at 660 nm) was added to each well. -2light), CPT, CPT-ss-PEG, (With / without 660 nm 0.5 W cm -2 light) 100 μL, respectively, and continue to culture for 24 h; add 50 μL of MTT, respectively, and continue to culture for 4 h in a 5% CO2, 37 °C incubator; discard the culture medium, add 150 μL of DMSO, shake well on a plate shaker, and read the plate on a microplate reader at 495 nm; calculate the cell inhibition rate according to the measured absorbance value as follows: Figure 9 a, irradiate the cells without drugs with a 660 nm laser (0.8 W cm -2 ) for 5 min, and the cells can still maintain a high survival rate, indicating that the 660 nm laser has good safety. In the β-CD-BDP group, even if the drug concentration reaches 100 μM under dark conditions, the cells still have a survival rate of 60%, indicating that the dark toxicity of β-CD-BDP is small. However, when irradiated with a laser for 5 min, 25 μM of β-CD-BDP can induce a large amount of cell death. When the concentration reaches 50 μM, the cell survival rate is close to 0, indicating that the photothermal efficiency of β-CD-BDP can still be highly maintained under physiological conditions. As shown in Figure 9 b, when a smaller power of 0.5 W cm -2 is used, we can still obtain similar changes in cell survival rate, indicating that β-CD-BDP indeed has good photothermal treatment effect in vitro. Figure 9 d, the cytotoxicity of β-CD-BDP is significantly enhanced after complexing with CPT-ss-PEG. For example, compared with Figure 9 b, the cell survival rate of 50 μM and 100 μM of β-CD-BDP is reduced by half, proving that β-CD-BDP@CPT-ss-PEG has good photothermal-chemotherapy synergistic effect.

[0091] Example 11: In vivo anti-tumor evaluation of

[0092] After Balb / c mice were implanted with 4T1 tumors on the abdomen (when the tumor size grew to about 100 mm 3 ), the mice were randomly divided into groups, groups, and PBS groups for anti-tumor treatment research (drug concentration was 200 μM, in situ treatment of mice with drugs, 660 nm laser irradiation was performed 10 h after administration, 0.4 W cm -2 light for 5 min, treatment was performed once every 3 days, and treatment was performed 3 times. As shown in Figure 10 a, during the treatment period, no obvious change in the body weight of the mice occurred, indicating that β-CD-BDP has little systemic toxicity. As Figure 10 As shown in b and c, The tumor volume of mice in the group was significantly reduced. The tumors in the PBS group did not increase significantly, while those in the PBS group increased significantly, indicating that the synergistic effect of photothermal and chemotherapy with #imgpt61# can effectively inhibit tumor growth. After treatment, the mice were dissected and their hearts, livers, spleens, lungs, kidneys, and tumors were weighed. Figure 10 As shown in Figure d, the tumors in the treated group were significantly smaller than those in the control group. Notably, the spleens in the treated group were also significantly smaller than those in the control group. This may be due to immune system disruption in the untreated control mice, leading to spleen enlargement.

Claims

1. A complex β-CD-BDP CPT-ss-PEG, characterized by: The complex β-CD-BDP CPT-ss-PEG is formed by complexing a compound represented by formula A and a compound represented by formula B. The method comprises: adding the compound represented by formula A (β-CD-BDP) and the compound represented by formula B (CPT-ss-PEG) into a solvent, dispersing them uniformly to cause complexation, and subjecting the resulting mixed solution to post-treatment H to obtain the complex β-CD-BDP. CPT-ss-PEG; β-CD-BDP CPT-ss-PEG A B In formula B, n=100-10000.

2. The complex β-CD-BDP according to claim 1 The preparation method of CPT-ss-PEG is characterized in that The method is: The compound of formula A and the compound of formula B are added to the solvent and dispersed evenly. The resulting mixture is subjected to post-treatment H to obtain the complex β-CD-BDP. CPT-ss-PEG; the molar ratio of the compound of formula A to the compound of formula B is 1:1; A B In formula B, n=100-10000.

3. The complex β-CD-BDP according to claim 2 The preparation method of CPT-ss-PEG is characterized by: The solvent is dimethyl sulfoxide, tetrahydrofuran, dimethylformamide or water.

4. The complex β-CD-BDP according to claim 2 The preparation method of CPT-ss-PEG is characterized by: The volume of the solvent is 5-50 mL / g based on the total mass of the compound of formula A and the compound of formula B.

5. The complex β-CD-BDP according to claim 2 The preparation method of CPT-ss-PEG is characterized in that The post-treatment H is: freeze-drying the mixed solution to obtain the complex β-CD-BDP CPT-ss-PEG.

6. The complex β-CD-BDP according to claim 2 The preparation method of CPT-ss-PEG is characterized in that The compound of formula A is synthesized as follows: (1) Preparation of OTs-CD: β-cyclodextrin was added to water to obtain a cyclodextrin solution, and an aqueous solution of NaOH was slowly added dropwise, followed by an acetonitrile solution of toluenesulfonyl chloride. After the addition was complete, the resulting mixed solution was stirred at room temperature for 2-5 h. The filtrate was collected, and the pH was adjusted to 7-8 with a saturated ammonium chloride solution. The precipitate was collected by centrifugation, and the precipitate was subjected to post-treatment A to obtain compound 1. The molar ratio of the β-cyclodextrin, the NaOH contained in the NaOH aqueous solution and the methylbenzenesulfonyl chloride contained in the acetonitrile solution of methylbenzenesulfonyl chloride is 1:1-10:1-5; 1 (2) Preparation of NH2-CD: Compound 1 described in step (1) is reacted with ethylenediamine at 50-90°C for 2-8 h, and the resulting reaction solution B is subjected to post-treatment B to obtain compound 2; The volume of the ethylenediamine is 2-10 mL / g based on the mass of the compound 1; 2 (3) Preparation of BDP-NHS: Compound 3 was dissolved in dimethylformamide A, followed by the addition of N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 4-dimethylaminopyridine. The mixture was reacted at room temperature for 12-36 h. After the reaction, the solvent was evaporated to dryness, and the crude product was washed with methanol to obtain compound 4. The molar ratio of the compound 3, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 4-dimethylaminopyridine is 1:1-5:1-5:1-3; 3 4 (4) Preparation of β-CD-BDP: Dissolve the compound 4 described in step (3) in dimethylformamide B, add the compound 2 described in step (2) and triethylamine, and react at room temperature for 12-36 hours. The resulting mixed solution is subjected to post-treatment D to obtain the compound of formula A; A The molar ratio of the compound 4, compound 2 and triethylamine is 1:0.2-1:0.1-0.

5.

7. The complex β-CD-BDP according to claim 6 The preparation method of CPT-ss-PEG is characterized by: The volume of water in step (1) is 5-10 mL / mmol based on the amount of β-cyclodextrin; The NaOH aqueous solution in step (1) is obtained by dissolving NaOH in water A, wherein the volume of water A is 0.1-0.2 mL / mmol based on the amount of NaOH substance; The acetonitrile solution of methylbenzenesulfonyl chloride in step (1) is obtained by dissolving methylbenzenesulfonyl chloride in acetonitrile, wherein the volume of acetonitrile is 0.1-0.3 mL / mmol based on the amount of methylbenzenesulfonyl chloride; The post-treatment A in step (1) is: washing the precipitate with water and acetone in sequence, and vacuum drying to obtain the compound represented by formula 1; The post-treatment B in step (2) is as follows: cooling the reaction solution B to room temperature, adding acetone A for precipitation, centrifuging, dissolving the resulting precipitate in water, re-precipitating with acetone B, centrifuging, and vacuum drying the resulting precipitate to obtain compound 2; The volume of dimethylformamide A in step (3) is 50-200 mL / g based on the mass of compound 3; The volume of dimethylformamide B in step (4) is 100-1000 mL / g based on the mass of compound 4; The post-treatment D in step (4) is: adding the mixed solution dropwise into acetone C for precipitation, centrifuging, washing the obtained precipitate with acetone, and vacuum drying to obtain the compound of formula A.

8. The complex β-CD-BDP according to claim 2 The preparation method of CPT-ss-PEG is characterized in that The compound of formula B is prepared as follows: Compound 5 is dissolved in dichloromethane, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 4-dimethylaminopyridine and compound 6 are added. The reaction is carried out at room temperature for 8-16 hours. The resulting reaction solution F is subjected to post-treatment F to obtain the compound of formula B; 5 6 B The molar ratio of the compound 5, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 4-dimethylaminopyridine and the compound 6 is 1:1-4:0.5-2:0.5-2; In Formula 5 and Formula B, n=100-10000.

9. The complex β-CD-BDP according to claim 8 The preparation method of CPT-ss-PEG is characterized in that The volume of the dichloromethane is 10-40 mL / g based on the mass of compound 5; The post-treatment F is as follows: adding water to the reaction solution F for extraction, taking the aqueous phase and adding dichloromethane for extraction, combining the organic phases, and drying to obtain the compound of formula B.

10. Use of the complex according to claim 1 in the preparation of anti-tumor phototherapy drugs.

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