A thio-ellipticine derivative, its preparation method and application

Abstract

The present application relates to a kind of thio camptothecin derivatives and its preparation method and application, belong to the technical field of pharmaceutical chemistry and antitumor drug.The thio camptothecin derivative has the following structural general formula, R is selected from-H or-C (=O)-CH2-X-CH2-C (=O)-O-R1;X is selected from-S-、-O- or-CH2-;R1 It is selected from C1-C6 alkyl or benzyl optionally substituted by 1- CF3 Or-CN.The thio camptothecin derivative of oxygen sulfur replacement strategy modification provided by the present application is controllable, structure stable, itself has high efficient low toxicity antitumor activity, after being combined with copper ion, stronger tumor inhibition effect can be realized through chemical kinetics therapy, solve the technical defects such as poor water solubility of camptothecin, lactone ring easy ring-opening inactivation, dose-limiting toxicity, etc., with drug development potential.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry and antitumor drug technology, specifically relating to a thiocamptothecin derivative, its preparation method, and its application. Background Technology

[0002] Camptothecin, a specific inhibitor of DNA topoisomerase I, is an important lead molecule in the development of antitumor drugs. However, it suffers from poor water solubility, easy ring-opening inactivation of the lactone ring, and high dose-limiting toxicity, which severely restricts its clinical application. Current technologies for structural modification of camptothecin mainly focus on side chain modification and oxygen-sulfur substitution. While these modification strategies have improved the instability of the camptothecin lactone ring and high dose-limiting toxicity to some extent, they have not fundamentally solved its water solubility problem. The tumor microenvironment is characterized by weak acidity, high glutathione, and high reactive oxygen species (ROS). Tumor cells are more sensitive to oxidative stress. Developing ROS-mediated therapy that utilizes these characteristics to respond to endogenous stimuli is an important direction for achieving highly effective and low-toxicity antitumor therapy. However, most existing camptothecin derivatives lack the ability to specifically respond to the tumor microenvironment and cannot fully utilize the redox characteristics of tumor tissue to achieve drug release and enhance tumor-suppressive effects. Summary of the Invention

[0003] To address the aforementioned deficiencies of camptothecin in existing technologies, this invention provides a thiocamptothecin derivative, its preparation method, and its applications. Specifically, it provides a novel thiocamptothecin derivative, its preparation method, and its antitumor applications. This derivative exhibits controllable synthesis, structural stability, and highly efficient, low-toxicity antitumor activity. When combined with copper ions, it can achieve a stronger tumor-suppressing effect through chemokinetic therapy, demonstrating potential for drug development.

[0004] In a first aspect, the present invention provides a thiocamptothecin derivative having the following general structural formula:

[0005] Wherein, R is selected from -H or -C(=O)-CH2-X-CH2-C(=O)-O-R1; X is selected from -S-, -O- or -CH2-; R1 is selected from C1-C6 alkyl or optionally benzyl substituted with one -CF3 or -CN.

[0006] Preferably, the thiocamptothecin derivative is one of the following compounds:

[0007] Secondly, the present invention provides a method for preparing the above-mentioned thiocamptothecin derivatives, comprising the following steps: (1) An alcohol compound R1-OH and an acid anhydride compound are esterified in dichloromethane in the presence of a catalyst to obtain intermediate A. Wherein, the acid anhydride compound is selected from glutaric anhydride or thioglycolic acid anhydride; R1 is selected from C1-C6 alkyl or optionally benzyl group substituted with one -CF3 or -CN group; X is selected from S or O. (2) Intermediate A is mixed with condensing agent and catalyst in dichloromethane to carry out an activation reaction to obtain an activated system; camptothecin is added to the activated system to carry out an esterification condensation reaction to obtain camptothecin derivative B. Wherein, R is selected from -H or -C(=O)-CH2-X-CH2-C(=O)-O-R1; X is selected from -S-, -O- or -CH2-; R1 is selected from C1-C6 alkyl or optionally benzyl substituted with one -CF3 or -CN; (3) Add an organic solvent to camptothecin derivative B or camptothecin, then add Lawson's reagent, and carry out a reflux reaction under nitrogen protection. Separate and purify to obtain the target thiocamptothecin derivative. or Wherein, R is selected from -H or -C(=O)-CH2-X-CH2-C(=O)-O-R1; X is selected from -S-, -O- or -CH2-; R1 is selected from C1-C6 alkyl or optionally benzyl substituted with one -CF3 or -CN; the organic solvent is xylene or dichloroethane.

[0008] Furthermore, the catalyst is 4-dimethylaminopyridine (DMAP).

[0009] Furthermore, the condensation reagent is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI·HCl).

[0010] Furthermore, in step (1), the molar ratio of the alcohol compound R1-OH to the acid anhydride compound is 1.5:1.

[0011] Furthermore, in step (2), the molar ratio of intermediate A to camptothecin is 3:1.

[0012] Furthermore, in step (3), when the organic solvent is xylene, the molar ratio of camptothecin derivative B to Lawson reagent is 1:8~16; when the organic solvent is dichloroethane, the molar ratio of camptothecin derivative B to Lawson reagent is 1:3.

[0013] Furthermore, in step (1), the esterification reaction is carried out at room temperature.

[0014] Furthermore, in step (2), the activation reaction condition is activation at room temperature for 30 min; the esterification condensation reaction condition is stirring at room temperature until the reaction is complete.

[0015] Furthermore, in step (3), the heating reflux reaction conditions are 1.5 to 3 h at 80 to 130 °C.

[0016] Furthermore, the purification method is one or both of column chromatography and preparative thin-layer chromatography.

[0017] Thirdly, the present invention provides a complex based on a thiocamptothecin derivative, wherein the complex is composed of the above-mentioned thiocamptothecin derivative and Cu. 2+ Coordination complexes formed through multidentate chelation coordination.

[0018] Fourthly, the present invention provides a method for preparing the above-mentioned complex based on thiocamptothecin derivatives: take the above-mentioned thiocamptothecin derivatives and divalent copper salts, and prepare equal volumes of solutions according to a molar ratio of 1:2; mix the two solutions in equal volumes, stir thoroughly at room temperature, let stand, and remove the solvent by natural drying to obtain the final product.

[0019] Fifthly, the present invention provides the use of the above-mentioned thiocamptothecin derivatives or the above-mentioned complexes based on thiocamptothecin derivatives in the preparation of antitumor drugs.

[0020] Furthermore, the tumor is one or both of lung cancer and colorectal cancer.

[0021] The thiocamptothecin derivatives provided by this invention exhibit inhibitory activity comparable to camptothecin against A549 and HCT116 tumor cells, with significantly reduced cytotoxicity to normal cells. They can catalyze the generation of hydroxyl radicals and efficiently consume GSH in a weakly acidic tumor microenvironment, and can be used to prepare low-toxicity, high-efficiency antitumor drugs, which are chemokinetic therapy (CDT) drugs. Furthermore, the complex formed by combining the thiocamptothecin derivatives with copper ions can further enhance the chemokinetic therapy effect.

[0022] The beneficial effects of this invention are: 1. Existing camptothecin derivatives lack redox-sensitive structures and cannot specifically recognize the tumor microenvironment. This invention modifies the camptothecin nucleus by oxygen-sulfur substitution, enabling it to directionally catalyze the generation of hydroxyl radicals under weakly acidic conditions in tumors, while efficiently consuming the highly expressed GSH in tumor cells.

[0023] 2. The thiocamptothecin derivative provided by this invention has comparable inhibitory activity against the proliferation of A549 lung cancer and HCT116 colorectal cancer cells to that of natural camptothecin, but significantly reduced toxicity to normal cells. Traditional camptothecin and its modified products lack cell selectivity and are prone to causing damage to normal tissues, exhibiting dose-limiting toxicity.

[0024] 3. The thiocamptothecin derivative provided by this invention contains thiocarbonyl and hydroxyl coordination sites, enabling it to react with Cu. 2+ By achieving a stable complexation at a 1:2 molar ratio, the resulting coordination complex can further enhance the production of hydroxyl radicals, strengthen the oxidative killing effect, and integrate chemotherapy with chemokinetics, thus expanding the drug's application value. Furthermore, this thiocamptothecin derivative retains the chemotherapeutic activity of camptothecin in inhibiting DNA topoisomerase, while also possessing redox-responsive CDT function, making it suitable for the preparation of low-toxicity antitumor drugs for lung cancer and colorectal cancer. Attached Figure Description

[0025] Figure 1 The formation of compound C10 at different pH values The performance test results of OH oxidation of TMB are shown in the following figures: a) color changes of the control group (left) and compound C10 (right) after co-incubation with H2O2 and TMB for 30 min, and color changes after termination with dilute H2SO4; b) UV absorption changes of compound C10 after co-incubation with H2O2 and TMB for 30 min; c) UV absorption spectrum over time.

[0026] Figure 2 The color changes after camptothecin was incubated with H2O2 and TMB for 30 min (left) and the color changes after the incubation was terminated with dilute H2SO4 (right).

[0027] Figure 3 The results of the GSH consumption test for compound C10 are shown below. In this test, a is the UV absorption spectrum of the DTNB product reduced by GSH from compound C10; b is the peak change at 412 nm in the UV absorption spectrum of DTNB from compound C10; and c is the peak change at 412 nm in the UV absorption spectrum of DTNB from compound C10 compared with camptothecin.

[0028] Figure 4 The results show the UV fluorescence changes of compounds C10 and D1 after the addition of copper ions, where a represents the reaction between compound C10 and Cu. 2+ UV titration spectrum; b is the reaction spectrum of compound C10 with Cu 2+ The fluorescence titration spectrum; c represents the reaction of compound D1 with Cu. 2+ UV titration spectrum; d represents the reaction of compound D1 with Cu 2+ The fluorescence titration spectrum.

[0029] Figure 5 The image shows the UV absorption spectrum of compound C10 bound to copper ions (a) and the results of spot analysis (b).

[0030] Figure 6 This is a fitted curve of the ultraviolet absorption of compound C10.

[0031] Figure 7 Compound C10, complex C10-Cu 2+ Cu 2+ The infrared spectrum.

[0032] Figure 8 The complex C10-Cu 2+ High-resolution mass spectrometry.

[0033] Figure 9 The complex C10-Cu 2+ The XPS spectra are shown, where a is the photoelectron spectrum and b is the fine scan peak of copper.

[0034] Figure 10 The complex C10-Cu 2+ Generation at different pH Performance test results of OH oxidation of TMB, where a represents the complex C10-Cu 2+ UV absorption spectra after co-incubation with H2O2 and TMB for 30 min; b is the control group and the complex C10-Cu 2+ Color changes after co-incubation with H2O2 and TMB for 30 min and color changes after termination with dilute H2SO4; c represents the color-changing principle of TMB.

[0035] Figure 11 Compound C10, complex C10-Cu 2+ UV absorption results after co-incubation with H2O2 and TMB for 30 min.

[0036] Figure 12 The complex C10-Cu 2+ Results of GSH consumption capacity test, where a represents the complex C10-Cu 2+ DTNB UV absorption spectrum; b is the C10-Cu complex. 2+ The peak change at 412 nm in the DTNB UV absorption spectrum; c represents the C10-Cu complex. 2+ Compared with C10: Changes in the peak at 412 nm in the UV absorption spectrum of DTNB.

[0037] Figure 13 Compound C10, complex C10-Cu 2+ The results of intracellular reactive oxygen species detection.

[0038] Figure 14 Control (negative control), compound C10, complex C10-Cu 2+ Fluorescence intensity results after incubation with DCFH-DA dye.

[0039] Figure 15 Compound C10, complex C10-Cu 2+ The results of mitochondrial membrane potential analysis of A549 cells after drug administration, where a) is fluorescence imaging of C10 after incubation with JC-1 staining; b) is fluorescence imaging of the C10-Cu complex after incubation with JC-1 staining. 2+ Fluorescence imaging after incubation.

[0040] Figure 16 Compound C10, complex C10-Cu 2+ The results of apoptosis detection. Detailed Implementation

[0041] The following non-limiting embodiments are intended to enable those skilled in the art to more fully understand the invention, but do not limit the invention in any way.

[0042] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; the reagents and materials described are commercially available unless otherwise specified.

[0043] Compounds C1-C10 and D1-D12 in the following examples were prepared according to the following process route:

[0044] The R is selected from -H or -C(=O)-CH2-X-CH2-C(=O)-O-R1; the X is selected from -S-, -O- or -CH2-; the R1 is selected from C1-C6 alkyl or optionally benzyl substituted with one -CF3 or -CN.

[0045] The chemical structural formulas of compounds C1-C10 and D1-D12 are as follows:

[0046] Example 1 The synthesis of compound C1 includes the following steps: (1) Take 1.000 g (0.0057 mol) of 3-(trifluoromethyl)benzyl alcohol in a 25 mL round-bottom flask, add 10 mL of dichloromethane to dissolve it, add 0.350 g (0.0028 mol) of 4-dimethylaminopyridine (DMAP) and 1.000 g (0.0087 mol) of glutaric anhydride, stir at room temperature for 2 h, and monitor by TLC. After the reactants have reacted completely, add saturated sodium bicarbonate aqueous solution to adjust the pH to about 10, add 50 mL of dichloromethane for extraction, combine the aqueous phases, slowly add concentrated hydrochloric acid to adjust the pH to about 3, a white precipitate precipitates, add 50 mL of dichloromethane for extraction twice, combine the organic phases, dry with anhydrous Na2SO4, distill under reduced pressure to obtain 1.488 g (5.1310 mmol) of dark brown oily liquid, R f =0.5 (dichloromethane:methanol = 30:1, V / V The yield was 90%, and it was denoted as intermediate A1.

[0047] The structural formula of intermediate A1 is .

[0048] (2) Intermediate A1 (187 mg, 0.65 mmol) was added to a round-bottom flask and dissolved in dichloromethane (DCM). EDC·HCl (330 mg, 1.72 mmol) and DMAP (105 mg, 0.86 mmol) were added sequentially, and the mixture was activated at room temperature for 30 min. Camptothecin (150 mg, 0.43 mmol) was then added, and the mixture was stirred overnight at room temperature. After the reaction was complete, the system was washed with water, the organic phase was separated and collected, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The crude product was purified by column chromatography (eluent: dichloromethane / methanol = 100~200:1) to obtain compound B1.

[0049] The structural formula of compound B1 is .

[0050] (3) Add 50 mg (0.08 mmol) of compound B1 to a round-bottom flask, then add 10 mL of xylene. Sonicate until completely dissolved, then add 535 mg (1.32 mmol) of Lawson's reagent. Add a stir bar, install a serpentine condenser, use a water pump to remove air, then introduce nitrogen gas, and sonicate again. Repeat three times to ensure no oxygen residue remains. Reflux the reaction and monitor the reaction by TLC until a large number of target spots are formed. Post-processing: After the reaction is complete, allow it to cool naturally to room temperature. Slurry the reaction solution three times with petroleum ether, filter to collect the reddish-brown precipitate, and purify it by preparative thin-layer chromatography to obtain compound C1. 5 mg (0.01 mmol) of reddish-brown solid was obtained, with a yield of 10%.

[0051] 1H NMR (400 MHz, DMSO- d 6) δ 7.87 (s, 1H), 7.26 (t, J = 9.2 Hz, 2H), 6.99(t, J = 8.5 Hz, 1H), 6.85 (d, J = 6.8 Hz, 1H), 6.81 (d, J = 8.3 Hz, 2H), 6.72 (s,1H), 6.70 (s, 2H), 5.00 (d, J = 17.1 Hz, 1H), 4.75 (d, J = 4.0 Hz, 1H), 4.73 –4.70 (m, 2H), 4.64 (s, 2H), 2.02 – 1.96 (m, 2H), 1.80 (t, J = 7.9 Hz, 2H), 1.29(q, J = 7.4 Hz, 2H), 1.14 (t, J = 7.3 Hz, 2H), 0.06–0.00 (m, 3H). HRMS(ESI): m / z:[M+H] + : calcd for C 33 H 28 F3N2O5S2653.1392; found: 653.1395. Example 2 The synthesis of compound C2 includes the following steps: (1) Take 1.000 g (0.0057 mol) of 3-(trifluoromethyl)benzyl alcohol in a 25 mL round-bottom flask, add 10 mL of dichloromethane to dissolve it, add 0.350 g (0.0028 mol) of 4-dimethylaminopyridine (DMAP) and 1.000 g (0.0087 mol) of glutaric anhydride, stir at room temperature for 2 h, and monitor by TLC. After the reactants have reacted completely, add saturated sodium bicarbonate aqueous solution to adjust the pH to about 10, add 50 mL of dichloromethane for extraction, combine the aqueous phases, slowly add concentrated hydrochloric acid to adjust the pH to about 3, a white precipitate precipitates, add 50 mL of dichloromethane for extraction twice, combine the organic phases, dry with anhydrous Na2SO4, distill under reduced pressure to obtain 1.471 g (5.0720 mmol) of dark brown oily liquid, R f = 0.4 (dichloromethane:methanol = 30:1, V / V The yield was 89%, and it was denoted as intermediate A2.

[0052] The structural formula of intermediate A2 is .

[0053] (2) Using intermediate A2 instead of intermediate A1, compound B2 was prepared according to step (2) of Example 1.

[0054] The structural formula of compound B2 is: .

[0055] (3) Using compound B2 instead of compound B1 as the raw material, compound C2 was prepared according to step (3) of Example 1, with a yield of 10%.

[0056] 1H NMR (400 MHz, CDCl3) δ 15.65 (s, 1H), 15.42 (d, J = 8.3 Hz, 1H), 15.17 (d, J = 8.3 Hz, 1H), 15.05 (t, J = 7.7 Hz, 1H), 14.89 (t, J = 7.5 Hz,1H), 14.83 (s, 1H), 14.76 (d, J = 6.8 Hz, 2H), 14.72 (d, J = 7.9 Hz, 1H), 14.65 (t, J = 8.1 Hz, 1H), 13.18 (d, J = 17.6 Hz, 1H), 12.79 (d, J = 20.2 Hz,1H), 12.73 (d, J = 8.9 Hz, 2H), 12.68 (d, J = 6.0 Hz, 2H), 10.04 (t, J = 7.5Hz, 2H), 9.81 (td, J = 16.5, 9.2 Hz, 2H), 9.43 (ddd, J = 22.0, 14.2, 7.0 Hz, 2H), 9.34–9.27 (m, 2H), 8.15 (t, J = 7.5 Hz, 3H). HRMS(ESI): m / z: [M+H]+ :calcd for C 33 H 28 F3N2O5S2653.1392; found: 653.1395. Example 3 The synthesis of compound C3 includes the following steps: (1) Take 1.000 g (0.0057 mol) of 4-(trifluoromethyl)benzyl alcohol in a 25 mL round-bottom flask, add 10 mL of dichloromethane to dissolve it, add 0.350 g (0.0028 mol) of 4-dimethylaminopyridine (DMAP) and 1.200 g (0.0089 mol) of thioglycolic acid anhydride, stir at room temperature for 2 h, and monitor by TLC. After the reactants have reacted completely, add saturated sodium bicarbonate aqueous solution to adjust the pH to about 10, add 50 mL of dichloromethane for extraction, combine the aqueous phases, slowly add concentrated hydrochloric acid to adjust the pH to about 3, a white precipitate precipitates, add 50 mL of dichloromethane for extraction twice, combine the organic phases, dry with anhydrous Na2SO4, distill under reduced pressure to obtain 1.509 g (4.8990 mmol) of dark brown oily liquid, R f =0.3 (dichloromethane:methanol = 30:1, V / V The yield was 86%, and it was denoted as intermediate A3.

[0057] The structural formula of intermediate A3 is .

[0058] (2) Using intermediate A3 instead of intermediate A1, compound B3 was prepared according to step (2) of Example 1.

[0059] The structural formula of compound B3 is: .

[0060] (3) Using compound B3 instead of compound B1 as the raw material, compound C3 was prepared according to step (3) of Example 1, with a yield of 9%.

[0061] 1H NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 7.33 (d, J = 8.8 Hz, 2H), 7.04 (t, J = 7.6 Hz, 1H), 6.92 (d, J = 8.3 Hz, 2H), 6.88 (s, 1H), 6.85 (s,1H), 6.80 (d, J = 7.9 Hz, 2H), 5.07 (d, J = 17.1 Hz, 1H), 4.81 (d, J = 9.0Hz, 1H), 4.80 – 4.75 (m, 2H), 4.71 (s, 2H), 3.11 (s, 2H), 3.05 – 2.93 (m,2H), 1.31 (q, J = 7.6 Hz, 2H), 0.07 (t, J = 7.3 Hz, 3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 32 H26 F3N2O5S3671.0956; found: 671.0958 . Example 4 The synthesis of compound C4 includes the following steps: (1) Take 1.000 g (0.0057 mol) of 4-(trifluoromethyl)benzyl alcohol in a 25 mL round-bottom flask, add 10 mL of dichloromethane to dissolve it, add 0.350 g (0.0028 mol) of 4-dimethylaminopyridine (DMAP) and 1.200 g (0.0089 mol) of thioglycolic acid anhydride, stir at room temperature for 2 h, and monitor by TLC. After the reactants have reacted completely, add saturated sodium bicarbonate aqueous solution to adjust the pH to about 10, add 50 mL of dichloromethane for extraction, combine the aqueous phases, slowly add concentrated hydrochloric acid to adjust the pH to about 3, a white precipitate precipitates, add 50 mL of dichloromethane for extraction twice, combine the organic phases, dry with anhydrous Na2SO4, distill under reduced pressure to obtain 1.509 g (4.8990 mmol) of dark brown oily liquid, R f =0.3 (dichloromethane:methanol = 30:1, V / V The yield was 86%, and it was denoted as intermediate A4.

[0062] The structural formula of intermediate A4 is .

[0063] (2) Using intermediate A4 instead of intermediate A1, compound B4 was prepared according to step (2) of Example 1 to obtain compound B4.

[0064] The structural formula of compound B4 is: .

[0065] (3) Using compound B4 instead of compound B1 as the raw material, compound C4 was prepared according to step (3) of Example 1, with a yield of 10%.

[0066] 1H NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 7.32 (d, J = 8.4 Hz, 2H), 7.04 (t, J = 8.5 Hz, 1H), 6.98 (s, 1H), 6.92 (d, J = 8.2 Hz, 1H), 6.87 (d, J = 9.5 Hz, 2H), 6.85 (s, 1H), 6.77 (t, J = 7.6 Hz, 1H), 5.06 (d, J = 17.0 Hz, 1H), 4.81 (d, J = 6.7 Hz, 1H), 4.79 – 4.75 (m, 2H), 4.70 (s, 2H), 3.11 (s, 2H), 3.03 – 2.92 (m, 2H), 1.31 (q, J = 7.3 Hz, 2H), 0.07 (t, J = 7.3 Hz, 3H).HRMS(ESI): m / z: [M+H]+ : calcd for C 32 H 26 F3N2O5S3671.0956; found: 671.0958 . Example 5 The synthesis of compound C5 includes the following steps: (1) Take 1.000 g (0.0057 mol) of 3-(trifluoromethyl)benzyl alcohol in a 25 mL round-bottom flask, add 10 mL of dichloromethane to dissolve it, add 0.350 g (0.0028 mol) of 4-dimethylaminopyridine (DMAP) and 1.050 g (0.0073 mol) of diethylene glycol anhydride, stir at room temperature for 2 h, and monitor by TLC. After the reactants have reacted completely, add saturated sodium bicarbonate aqueous solution to adjust the pH to about 10, add 50 mL of dichloromethane for extraction, combine the aqueous phases, slowly add concentrated hydrochloric acid to adjust the pH to about 3, a white precipitate will precipitate, add 50 mL of dichloromethane to extract twice, combine the organic phases, dry with anhydrous Na2SO4, distill under reduced pressure to obtain 1.515 g (5.1883 mmol) of dark brown oily liquid, R f =0.4 (dichloromethane:methanol = 30:1, V / V The yield was 91%, and it was designated as intermediate A5.

[0067] The structural formula of intermediate A5 is .

[0068] (2) Using intermediate A5 instead of intermediate A1, compound B5 was prepared according to step (2) of Example 1.

[0069] The structural formula of compound B5 is: .

[0070] (3) Using compound B5 instead of compound B1 as the raw material, compound C5 was prepared according to step (3) of Example 1, with a yield of 11%.

[0071] 1H NMR (400 MHz, DMSO-d6) δ 7.30 (s, 1H), 6.72 – 6.67 (m, 2H), 6.44 –6.38 (m, 1H), 6.29 (d, J = 4.5 Hz, 1H), 6.27 (d, J = 4.8 Hz, 2H), 6.23 (s,1H), 6.18 (d, J = 8.3 Hz, 2H), 4.45 (d, J = 17.0 Hz, 1H), 4.20 (d, J = 8.6Hz, 1H), 4.18 – 4.14 (m, 2H), 4.07 (s, 2H), 2.43 – 2.31 (m, 2H), 1.92 (s, 2H), 0.70 (q, J = 7.3 Hz, 2H), -0.54 (t, J = 7.4 Hz, 3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 32 H 26 F3N2O6S2655.1184; found: 655.1179. Example 6 Synthesis of compound C6 (1) Diethylene glycol anhydride was placed in a round-bottom flask, and methanol and 4-dimethylaminopyridine (DMAP) were added. The mixture was stirred at room temperature for 2 h and monitored by TLC. After the reaction of the raw materials was complete, saturated sodium bicarbonate aqueous solution was added to adjust the pH to about 10. 50 mL of dichloromethane was added for extraction. The aqueous phases were combined, and concentrated hydrochloric acid was slowly added dropwise to adjust the pH to about 3. A white precipitate was formed. 50 mL of dichloromethane was added for extraction twice. The organic phases were combined, dried over anhydrous Na2SO4, and distilled under reduced pressure to obtain a dark brown oily liquid. R f =0.4 (dichloromethane:methanol = 30:1, V / V ), denoted as intermediate 1.

[0072] The structural formula of intermediate 1 is .

[0073] (2) Using intermediate 1 instead of intermediate A1, compound B9 was prepared according to step (2) of Example 1.

[0074] The structural formula of compound B9 is: .

[0075] (3) Compound C6 was prepared by replacing compound B1 with compound B9 as a raw material according to the method of Example 1, with a yield of 13%.

[0076] 1H NMR (400 MHz, DMSO-d6) δ 7.87 (s, 1H), 7.29 (d, J = 8.4 Hz, 1H), 7.25 (d, J = 8.3 Hz, 1H), 6.99 (t, J = 7.0 Hz, 1H), 6.84 (t, J = 7.6 Hz, 1H), 6.68 (s, 1H), 4.96 (d, J = 17.1 Hz, 1H), 4.69 (d, J = 17.0 Hz, 1H), 4.62 (s, 2H), 2.66 (s, 3H), 1.72 (t, J = 7.4 Hz, 2H), 1.49 (t, J = 7.5 Hz, 2H), 1.28(q, J = 7.5 Hz, 2H), 0.92 (q, J = 7.3 Hz, 2H), 0.00 (d, J = 7.6 Hz, 3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 26 H 25 N2O5S2509.1205; found: 509.1208. Example 7 Synthesis of compound C7 (1) In a round-bottom flask containing pentanol, dichloromethane was added to dissolve the reactants. Then, 4-dimethylaminopyridine (DMAP) and diethylene glycol anhydride were added, and the mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After the reactants had reacted completely, saturated sodium bicarbonate aqueous solution was added to adjust the pH to about 10. 50 mL of dichloromethane was added for extraction. The aqueous phases were combined, and concentrated hydrochloric acid was slowly added dropwise to adjust the pH to about 3. A white precipitate formed. 50 mL of dichloromethane was added for extraction twice. The organic phases were combined, dried over anhydrous Na2SO4, and distilled under reduced pressure to obtain a dark brown oily liquid. R f =0.4 (dichloromethane:methanol = 30:1, V / V ), denoted as intermediate 2.

[0077] The structural formula of intermediate 2 is .

[0078] (2) Using intermediate 2 instead of intermediate A1, compound B10 was prepared according to step (2) of Example 1.

[0079] The structural formula of compound B10 is .

[0080] (3) Compound C7 was prepared by replacing compound B1 with compound B10 as a raw material according to the method of Example 1, with a yield of 12%.

[0081] 1H NMR (400 MHz, DMSO-d6) δ 7.89 (s, 1H), 7.28 (t, J = 8.6 Hz, 2H), 7.03 – 6.98 (m, 1H), 6.86 (t, J = 7.5 Hz, 1H), 6.70 (s, 1H), 4.99 (d, J =17.1 Hz, 1H), 4.71 (d, J = 17.0 Hz, 1H), 4.65 (s, 2H), 3.51 (tt, J = 6.5, 3.9Hz, 2H), 2.92 – 2.88 (m, 2H), 1.89 (t, J = 7.6 Hz, 2H), 1.76 (t, J = 7.3 Hz,2H), 1.30 (q, J = 7.4 Hz, 2H), 1.08 (q, J = 7.0 Hz, 2H), 0.76 (q, J = 6.9 Hz,2H), 0.37 – 0.34 (m, 2H), 0.04 (t, J = 7.4 Hz, 3H), -0.08 (t, J = 7.2 Hz,3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 30 H 33 N2O5S2565.1831; found: 565.1829. Example 8 Synthesis of compound C8 (1) Thioglycolic acid anhydride was placed in a round-bottom flask, and methanol and 4-dimethylaminopyridine (DMAP) were added. The mixture was stirred at room temperature for 2 h and monitored by TLC. After the reaction of the raw materials was complete, saturated sodium bicarbonate aqueous solution was added to adjust the pH to about 10. 50 mL of dichloromethane was added for extraction. The aqueous phases were combined, and concentrated hydrochloric acid was slowly added dropwise to adjust the pH to about 3. A white precipitate was formed. 50 mL of dichloromethane was added for extraction twice. The organic phases were combined, dried over anhydrous Na2SO4, and distilled under reduced pressure to obtain a dark brown oily liquid. R f =0.4 (dichloromethane:methanol = 30:1, V / V ), denoted as intermediate 3.

[0082] The structural formula of intermediate 3 is .

[0083] (2) Intermediate 3 was used instead of intermediate A1 and compound B11 was prepared according to step (2) of Example 1.

[0084] The structural formula of compound B11 is .

[0085] (3) Using compound B11 instead of compound B1 as the raw material, compound C8 was prepared according to the method of Example 1, with a yield of 10%.

[0086] 1H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.34 (t, J = 7.6 Hz, 2H), 7.06 (t, J = 8.4 Hz, 1H), 6.92 (d, J = 8.2 Hz, 1H), 6.88 (s, 1H), 5.07 (d, J = 17.0 Hz, 1H), 4.78 (d, J = 17.0 Hz, 1H), 4.71 (s, 2H), 3.20 (d, J = 2.8 Hz, 2H), 3.05 (d, J = 2.9 Hz, 2H), 1.41 – 1.33 (m, 2H), 0.15 – 0.08 (m, 3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 25 H 23 N2O5S3527.0769; found: 527.0771. Example 9 Synthesis of compound C9 (1) Take pentanol in a round-bottom flask, add dichloromethane to dissolve it, add 4-dimethylaminopyridine (DMAP) and thioglycolic acid anhydride, stir at room temperature for 2 h, and monitor by TLC. After the reactants have reacted completely, add saturated sodium bicarbonate aqueous solution to adjust the pH to about 10, add 50 mL of dichloromethane for extraction, combine the aqueous phases, slowly add concentrated hydrochloric acid to adjust the pH to about 3, a white precipitate will precipitate, add 50 mL of dichloromethane to extract twice, combine the organic phases, dry with anhydrous Na2SO4, distill under reduced pressure to obtain a dark brown oily liquid, R f =0.4 (dichloromethane:methanol = 30:1, V / V ), denoted as intermediate 4.

[0087] The structural formula of intermediate 4 is .

[0088] (2) Using intermediate 4 instead of intermediate A1, compound B12 was prepared according to step (2) of Example 1.

[0089] The structural formula of compound B12 is .

[0090] (3) Compound C9 was prepared by replacing compound B1 with compound B12 as a raw material according to the method of Example 1, with a yield of 11%.

[0091] 1H NMR (400 MHz, DMSO-d6) δ 7.97 (s, 1H), 7.35 (d, J = 9.7 Hz, 2H), 7.08 (t, J = 8.4 Hz, 1H), 6.93 (t, J = 7.5 Hz, 1H), 6.90 (s, 1H), 5.09 (d, J= 17.0 Hz, 1H), 4.80 (d, J = 17.0 Hz, 1H), 4.74 (s, 2H), 3.58 (q, J = 6.5 Hz,2H), 3.04 (s, 2H), 3.02 – 2.92 (m, 2H), 1.39 (q, J = 7.3 Hz, 2H), 0.86 (t, J= 7.0 Hz, 2H), 0.46 (d, J = 3.9 Hz, 2H), 0.41 (s, 2H), 0.13 (t, J = 7.4 Hz, 3H), 0.03 – -0.01 (m, 3H). HRMS(ESI): m / z: [M+H]+ : calcd forC 29 H 31 N2O5S3583.1395; found: 583.1398. Example 10 Synthesis of compound C10 Add 50 mg (0.14 mmol) of camptothecin to a round-bottom flask, then add 10 mL of xylene. Sonicate until completely dissolved, then add 464 mg (1.15 mmol) of Lawson's reagent. Add a stir bar, install a serpentine condenser, evacuate the air using a water pump, then purge with nitrogen. Repeat the sonication process three times to ensure no oxygen residue remains. Reflux the reaction and monitor the reaction by TLC until a large number of target spots are formed. Post-processing: After the reaction, allow it to cool naturally to room temperature. Pulverize the reaction solution three times with petroleum ether, filter to collect the reddish-brown precipitate, and purify by preparative thin-layer chromatography to obtain compound C10. 18 mg (0.05 mmol) of reddish-brown solid was obtained, with a yield of 33%.

[0092] 1H NMR (600 MHz, DMSO-d6) δ 7.93 (s, 1H), 7.34 (d, J = 4.3 Hz, 1H), 7.31 (d, J = 3.9 Hz, 1H), 7.04 (t, J = 6.9 Hz, 1H), 6.94 (s, 1H), 6.90 (t, J= 7.5 Hz, 1H), 4.89 (d, J = 16.0 Hz, 1H), 4.76 (d, J = 16.0 Hz, 1H), 4.68 (s,2H), 1.19 (dd, J = 13.6, 7.0 Hz, 2H), 0.16 (t, J = 7.4 Hz, 3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 20 H 17 N2O2S2381.0731; found: 381.0723. Example 11 Synthesis of compound D1 In a round-bottom flask, 50 mg (0.08 mmol) of compound B1 obtained in Example 1 was added, followed by 10 mL of dichloroethane. The mixture was sonicated until completely dissolved, and 134 mg (0.33 mmol) of Lawson's reagent was added. A stir bar was added, a serpentine condenser was installed, air was evacuated using a water pump, and nitrogen gas was introduced. The mixture was then sonicated and repeated three times to ensure no oxygen residue remained. The reaction was refluxed, and the reaction was monitored by TLC until a large number of target spots were formed. Post-processing: After the reaction was complete, the mixture was allowed to cool naturally to room temperature. The reaction solution was slurried three times with petroleum ether, filtered, and the reddish-brown precipitate was collected. After purification by preparative thin-layer chromatography, compound D1 was obtained. 28 mg (0.04 mmol) of reddish-brown solid was obtained, with a yield of 55%.

[0093] 1H NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 7.33 (t, J = 9.4 Hz, 2H), 7.04 (t, J = 8.4 Hz, 1H), 6.91 (t, J = 7.5 Hz, 1H), 6.85 (d, J = 8.2 Hz, 2H), 6.76 (s, 1H), 6.70 (d, J = 8.2 Hz, 2H), 5.05 (d, J = 17.1 Hz, 1H), 4.78 (d, J= 17.0 Hz, 1H), 4.71 (s, 2H), 4.36 (s, 2H), 1.82 (t, J = 7.3 Hz, 2H), 1.36(q, J = 7.2 Hz, 2H), 1.04 (q, J = 7.3 Hz, 2H), 0.40 (s, 2H), 0.11 – 0.06 (m,3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 33 H 28 F3N2O6S 637.1620; found:637.1622. Example 12 Synthesis of compound D2 Using compound B2 obtained in Example 2 instead of compound B1 as the starting material, compound D2 was prepared according to the method in Example 11, with a yield of 54%.

[0094] 1H NMR (400 MHz, DMSO-d6) 6.89 (s, 1H), 6.87 – 6.81 (m, 2H), 6.77 (d, J = 5.7 Hz, 2H), 5.07 (d, J = 17.0 Hz, 1H), 4.80 (d, J = 17.1 Hz, 1H), 4.74 (s, 2H), 4.38 (s, 2H), 1.83 (t, J = 7.3 Hz, 2H), 1.37 (q, J = 7.6Hz, 2H), 1.05 (p, J = 7.1 Hz, 2H), 0.43 (s, 2H), 0.13 – 0.07 (m, 3H). HRMS(ESI): m / z: [M+H]+ : calcd for C33 H 28 F3N2O6S 637.1620; found: 637.1616. Example 13 Synthesis of compound D3 Using compound B3 obtained in Example 3 instead of compound B1 as the starting material, compound D3 was prepared according to the method in Example 11, with a yield of 51%.

[0095] 1H NMR (400 MHz, DMSO-d6) 6.86 (s, 1H), 6.83 (d, J= 7.9 Hz, 2H), 6.68 (d, J = 8.2 Hz, 2H), 5.05 (d, J = 16.9 Hz, 1H), 4.77 (d,J = 17.0 Hz, 1H), 4.69 (s, 2H), 4.36 (s, 2H), 2.96 – 2.87 (m, 2H), 2.75 (s, 2H), 1.31 (q, J = 7.4 Hz, 2H), 0.06 (d, J = 7.6 Hz, 3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 32 H 26 F3N2O6S2655.1184; found: 655.1179. Example 14 Synthesis of compound D4 Using compound B4 obtained in Example 4 instead of compound B1 as the starting material, compound D4 was prepared according to the method in Example 11, with a yield of 52%.

[0096] 1H NMR (400 MHz, DMSO-d6) δ 7.96 (s, 1H), 7.34 (dd, J = 8.4, 4.8 Hz, 2H), 7.05 (t, J = 7.4 Hz, 1H), 6.93 (d, J = 8.1 Hz, 1H), 6.88 (s, 2H), 6.82(dd, J = 15.5, 8.0 Hz, 3H), 6.74 (t, J = 7.5 Hz, 1H), 5.08 (d, J = 16.9 Hz,1H), 4.79 (d, J = 17.1 Hz, 1H), 4.73 (s, 2H), 4.40 (t, J = 4.4 Hz, 2H), 2.93(d, J = 7.2 Hz, 2H), 2.77 (s, 2H), 1.34 (q, J = 7.0 Hz, 2H), 0.09 (t, J = 7.4Hz, 3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 32 H 26 F3N2O6S2655.1184; found:655.1186. Example 15 Synthesis of compound D5 Using compound B5 obtained in Example 5 instead of compound B1 as the starting material, compound D5 was prepared according to the method in Example 11, with a yield of 53%.

[0097] 1H NMR (400 MHz, DMSO-d6) δ 7.98 (s, 1H), 7.37 (t, J = 8.4 Hz, 2H), 7.07 (t, J = 7.8 Hz, 1H), 6.94 (t, J = 7.5 Hz, 1H), 6.90 – 6.87 (m, 3H), 6.76(d, J = 8.1 Hz, 2H), 5.11 (d, J = 17.0 Hz, 1H), 4.82 (d, J = 16.8 Hz, 1H), 4.75 (s, 2H), 4.46 (s, 2H), 3.88 (d, J = 17.1 Hz, 1H), 3.68 (d, J = 17.1 Hz,1H), 3.54 (s, 2H), 1.39 (q, J = 7.7 Hz, 2H), 0.11 (t, J = 7.7 Hz, 3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 32 H 26 F3N2O7S 639.1413; found: 639.1417. Example 16 The synthesis of compound D6 includes the following steps: (1) Take 1.000 g (0.0057 mol) of 4-(trifluoromethyl)benzyl alcohol in a 25 mL round-bottom flask, add 10 mL of dichloromethane to dissolve it, add 0.350 g (0.0028 mol) of 4-dimethylaminopyridine (DMAP) and 1.050 g (0.0073 mol) of diethylene glycol anhydride, stir at room temperature for 2 h, and monitor by TLC. After the reactants have reacted completely, add saturated sodium bicarbonate aqueous solution to adjust the pH to about 10, add 50 mL of dichloromethane for extraction, combine the aqueous phases, slowly add concentrated hydrochloric acid to adjust the pH to about 3, a white precipitate precipitates, add 50 mL of dichloromethane for extraction twice, combine the organic phases, dry with anhydrous Na2SO4, distill under reduced pressure to obtain 1.498 g (5.1301 mmol) of dark brown oily liquid, R f =0.3 (dichloromethane:methanol = 30:1, V / V The yield was 90%, and it was designated as intermediate A6.

[0098] The structural formula of intermediate A6 is .

[0099] (2) Using intermediate A6 instead of intermediate A1, compound B6 was prepared according to step (2) of Example 1 to obtain compound B6.

[0100] The structural formula of compound B6 is: .

[0101] (3) Compound D6 was prepared by replacing compound B1 with compound B6 as the raw material according to the method of Example 11, with a yield of 56%.

[0102] 1H NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 7.33 (t, J = 8.9 Hz, 2H), 7.03 (t, J = 8.4 Hz, 1H), 6.92 – 6.86 (m, 2H), 6.86 – 6.78 (m, 3H), 6.73 (t,J = 7.7 Hz, 1H), 5.08 (d, J = 17.0 Hz, 1H), 4.78 (d, J = 17.1 Hz, 1H), 4.70(s, 2H), 4.42 (s, 2H), 3.84 (d, J = 17.1 Hz, 1H), 3.65 (d, J = 17.0 Hz, 1H), 3.50 (s, 2H), 1.35 (q, J = 7.2 Hz, 2H), 0.11 – 0.05 (m, 3H). HRMS(ESI): m / z:[M+H]+ : calcd for C 32 H 26 F3N2O7S 639.1413; found: 639.1416. Example 17 The synthesis of compound D7 includes the following steps: (1) Take 2.000 g (0.0076 mol) of 3-cyanobenzaldehyde in a 25 mL round-bottom flask, add 10 mL of methanol to dissolve it, add 2.520 g (0.0092 mol) of sodium borohydride, stir in an ice bath for 1.5 h, and monitor by TLC. After the reactants have reacted completely, extract the reaction solution three times with dichloromethane / water, combine the organic phases, dry with anhydrous Na2SO4, and distill under reduced pressure to obtain 1.588 g (0.0119 mol) of crude 3-hydroxymethylbenzonitrile, a pale yellow liquid, with a yield of 78%. 1.000 g (0.0057 mol) of 3-hydroxymethylbenzonitrile was dissolved in 10 mL of dichloromethane in a 25 mL round-bottom flask. Then, 0.350 g (0.0028 mol) of 4-dimethylpyridine (DMAP) and 1.280 g (0.0112 mol) of glutaric anhydride were added. The mixture was stirred at room temperature for 2 h and monitored by TLC. After the reactants had reacted completely, saturated sodium bicarbonate aqueous solution was added to adjust the pH to approximately 10. 50 mL of dichloromethane was added for extraction. The aqueous phases were combined, and concentrated hydrochloric acid was slowly added dropwise to adjust the pH to approximately 3. A white precipitate formed. The mixture was extracted twice with 50 mL of dichloromethane. The organic phases were combined, dried over anhydrous Na₂SO₄, and distilled under reduced pressure to obtain 1.211 g (4.9028 mmol) of a dark brown oily liquid. f =0.3 (dichloromethane:methanol = 30:1, V / V The yield was 86%, and it was designated as intermediate A7.

[0103] The structural formula of intermediate A7 is .

[0104] (2) Using intermediate A7 instead of intermediate A1, compound B7 was prepared according to step (2) of Example 1 to obtain compound B7.

[0105] The structural formula of compound B7 is: .

[0106] (3) Compound D7 was prepared by replacing compound B1 with compound B7 as a raw material according to the method of Example 11, with a yield of 53%.

[0107] 1H NMR (400 MHz, DMSO-d6) δ 7.97 (s, 1H), 7.35 (t, J = 7.9 Hz, 2H), 7.07 (t, J = 7.5 Hz, 1H), 6.94 (d, J = 8.8 Hz, 3H), 6.83 (d, J = 7.9 Hz, 1H), 6.78 (s, 1H), 6.72 (t, J = 7.6 Hz, 1H), 5.07 (d, J = 17.1 Hz, 1H), 4.80 (d, J= 17.1 Hz, 1H), 4.74 (s, 2H), 4.32 (s, 2H), 1.83 (t, J = 7.3 Hz, 2H), 1.38(q, J = 7.4 Hz, 2H), 1.06 (q, J = 7.3 Hz, 2H), 0.43 (s, 2H), 0.14 – 0.08 (m,3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 33 H 28 N3O6S 594.1699; found: 594.1703. Example 18 The synthesis of compound D8 includes the following steps: (1) Take 2.000 g (0.0076 mol) of 3-cyanobenzaldehyde in a 25 mL round-bottom flask, add 10 mL of methanol to dissolve it, add 2.520 g (0.0092 mol) of sodium borohydride, stir in an ice bath for 1.5 h, and monitor by TLC. After the reactants have reacted completely, extract the reaction solution three times with dichloromethane / water, combine the organic phases, dry with anhydrous Na2SO4, and distill under reduced pressure to obtain 1.588 g (0.0119 mol) of crude 3-hydroxymethylbenzonitrile, a pale yellow liquid, with a yield of 78%. 1.000 g (0.0057 mol) of 3-hydroxymethylbenzonitrile was dissolved in 10 mL of dichloromethane in a 25 mL round-bottom flask. Then, 0.350 g (0.0028 mol) of 4-dimethylaminopyridine (DMAP) and 1.509 g (0.0113 mol) of thioglycolic acid anhydride were added. The mixture was stirred at room temperature for 2 h and monitored by TLC. After the reactants had reacted completely, saturated sodium bicarbonate aqueous solution was added to adjust the pH to approximately 10. 50 mL of dichloromethane was added for extraction. The aqueous phases were combined, and concentrated hydrochloric acid was slowly added dropwise to adjust the pH to approximately 3. A white precipitate formed. The precipitate was extracted twice with 50 mL of dichloromethane. The organic phases were combined, dried over anhydrous Na₂SO₄, and distilled under reduced pressure to obtain 1.284 g (4.8453 mmol) of a dark brown oily liquid. f=0.3 (dichloromethane:methanol = 30:1, V / V The yield was 85%, and it was designated as intermediate A8.

[0108] The structural formula of intermediate A8 is .

[0109] (2) Using intermediate A8 instead of intermediate A1, compound B8 was prepared according to step (2) of Example 1 to obtain compound B8.

[0110] The structural formula of compound B8 is: .

[0111] (3) Compound D8 was prepared by replacing compound B1 with compound B8 as a raw material according to the method of Example 11, with a yield of 50%.

[0112] 1H NMR (400 MHz, DMSO-d6) δ 7.67 (s, 1H), 7.05 (t, J = 7.2 Hz, 2H), 6.77 (t, J = 7.6 Hz, 1H), 6.64 (s, 2H), 6.61 (s, 1H), 6.51 (t, J = 8.4 Hz,2H), 6.42 (t, J = 8.1 Hz, 1H), 4.80 (d, J = 17.0 Hz, 1H), 4.52 (d, J = 17.1Hz, 1H), 4.44 (s, 2H), 4.07 (d, J = 15.9 Hz, 2H), 2.68 (s, 2H), 2.39 (d, J =11.1 Hz, 2H), 1.08 (q, J = 7.2 Hz, 2H), -0.18 (t, J = 7.4 Hz, 3H). HRMS(ESI):m / z: [M+H]+ : calcd for C 32 H 26 N3O6S2612.1263; found: 612.1266. Example 19 Synthesis of compound D9 Compound D9 was prepared by replacing compound B1 with compound B9 as the starting material according to the method of Example 11, with a yield of 60%.

[0113] 1H NMR (400 MHz, DMSO-d6) δ 7.87 (s, 1H), 7.27 (dd, J = 15.1, 8.4 Hz, 2H), 6.99 (t, J = 8.5 Hz, 1H), 6.84 (t, J = 7.6 Hz, 1H), 6.68 (s, 1H), 4.96(d, J = 17.1 Hz, 1H), 4.69 (d, J = 17.0 Hz, 1H), 4.62 (s, 2H), 2.66 (s, 3H), 1.72 (t, J = 7.4 Hz, 2H), 1.49 (t, J = 7.5 Hz, 2H), 1.28 (q, J = 7.5 Hz, 2H),0.91 (t, J = 7.5 Hz, 2H), 0.01 (t, J = 7.5 Hz, 4H). HRMS(ESI): m / z: [M+H]+ :calcd for C 26 H 25 N2O6S 493.1433; found: 493.1437. Example 20 Synthesis of compound D10 Compound D10 was prepared by replacing compound B1 with compound B10 as a raw material according to the method of Example 11, with a yield of 59%.

[0114] 1H NMR (400 MHz, DMSO-d6) δ 7.91 (s, 1H), 7.36 – 7.26 (m, 2H), 7.01(d, J = 8.9 Hz, 1H), 6.87 (t, J = 7.6 Hz, 1H), 6.71 (s, 1H), 5.00 (d, J =17.0 Hz, 1H), 4.73 (d, J = 17.1 Hz, 1H), 4.66 (s, 2H), 3.10 (t, J = 6.7 Hz,2H), 2.93 – 2.81 (m, 6H), 1.75 (t, J = 7.3 Hz, 2H), 1.51 (t, J = 7.4 Hz, 2H),1.31 HRMS(ESI): m / z: [M+H]+ :calcd for C 30 H 33 N2O6S 549.2059; found: 549.2062. Example 21 Synthesis of compound D11 Compound D11 was prepared by replacing compound B1 with compound B1 as a raw material according to the method of Example 11, with a yield of 56%.

[0115] 1H NMR (400 MHz, DMSO-d6) δ 7.65 (s, 1H), 7.09 – 7.01 (m, 2H), 6.81 –6.75 (m, 1H), 6.66 – 6.59 (m, 2H), 4.80 (d, J = 17.0 Hz, 1H), 4.51 (d, J =17.0 Hz, 1H), 4.41 (s, 2H), 2.70 – 2.58 (m, 2H), 2.40 (s, 2H), 1.39 (s, 3H), 1.10 (q, J = 7.5 Hz, 2H), -0.16 (t, J = 7.3 Hz, 3H). HRMS(ESI): m / z: [M+H]+ :calcd for C 25 H 23N2O6S2511.0998; found:511.1000. Example 22 Synthesis of compound D12 Compound D12 was prepared by replacing compound B1 with compound B12 as a raw material according to the method of Example 11, with a yield of 54%.

[0116] 1H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.37 – 7.31 (m, 2H), 7.09 –7.03 (m, 1H), 6.92 (d, J = 8.2 Hz, 1H), 6.89 (d, J = 4.0 Hz, 1H), 5.07 (d, J = 17.0 Hz, 1H), 4.79 (d, J = 17.0 Hz, 1H), 4.71 (s, 2H), 3.17 (q, J = 6.6 Hz, 2H), 2.92 (q, J = 15.2 Hz, 2H), 2.65 (s, 2H), 1.37 (q, J = 7.5 Hz, 2H), 0.65(q, J = 6.7 Hz, 2H), 0.42 – 0.36 (m, 6H), 0.12 (t, J = 7.3 Hz, 3H), -0.00 – -0.05 (m, 3H). HRMS(ESI): m / z: [M+H]+ : calcd for C 29 H 31 N2O6S2567.1624; found:567.1627. Example 23 In vitro tumor proliferation inhibitory activity assay of compounds C1-C10 and D1-D12 The in vitro activity test methods are as follows, and the results are shown in Table 1. Among them, paclitaxel, a commonly used anti-tumor drug in clinical practice, is the positive control experimental group.

[0117] (1) In vitro screening test for antitumor activity - 1 Screening method: microculture tetrozolium (MTT) reduction method; cell line: human lung cancer cells (A549); treatment time: 48 h; calculation of the half-maximal inhibitory concentration (IC50) of each compound on tumor cell growth. 50 (%, μM).

[0118] (2) In vitro screening test for antitumor activity - 2 Screening method: microculture tetrozolium (MTT) reduction method; cell line: human colon cancer cells (HCT116); treatment time: 48 h; calculation of the half-maximal inhibitory concentration (IC50) of each compound on tumor cell growth. 50 (%, μM).

[0119] (3) In vitro screening test for antitumor activity - 3 Screening method: microculture tetrozolium (MTT) reduction method; cell line: normal human kidney cells (HEK293T); treatment time: 48 h; calculation of the half-maximal inhibitory concentration (IC50) of each compound against tumor cell growth. 50 (μM) Table 1. In vitro tumor proliferation inhibitory activity of compounds C1-C10 and D1-D12

[0120] Example 24 Formation of compounds C10, D1-D12, and camptothecin at different pH values Performance tests of OH oxidation of TMB, results are shown in [link to results]. Figure 1 , Figure 2 .

[0121] For compounds C10, D1-D12, The properties of OH were tested. After oxidation with •OH, a colorless and transparent solution of 3,3',5,5'-tetramethylbenzidine (TMB) showed a new UV absorption peak at 652 nm and the solution turned blue. The solution turned blue at pH 5.0 and 6.8, but the change was not obvious at pH 7.4. After adding sulfuric acid to stop the reaction, the solution turned yellow.

[0122] Example 25 The GSH consumption capacity tests of compounds C10 and D1-D12 are shown in the figure. Figure 3 .

[0123] The GSH response and consumption performance of compounds C10 and D1-D12 were tested: Taking compound C10 as an example, GSH was added to a PBS solution of C10 at pH 7.4, and the changes in the GSH reduction product of DTNB were tested. Over time, the UV absorption peak at 412 nm in C10 gradually decreased, while the GSH content in the control group remained almost unchanged. At 60 min, 20 mM compound C10 consumed 59% of GSH; at 180 min, it consumed 87% of GSH. Furthermore, the GSH consumption capacity of compound C10 far exceeded that of camptothecin.

[0124] Example 26 Compound C10 and Cu 2+ The ultraviolet titration and fluorescence titration spectra were studied, and the binding constants were fitted. The results are shown in […]. Figure 4 , 5 6.

[0125] The test concentration of compound C10 was fixed at 20 μM, and Cu was gradually added dropwise to dichloroethane. 2+ The solution was tested, and the ultraviolet absorption spectrum was obtained. A fluorescence titration experiment was performed on compound C10 at a concentration of 20 μM, and the fluorescence emission spectrum was obtained. The values ​​from the ultraviolet titration of compound C10 were used to fit an equation, and the reaction between compound C10 and Cu... 2+ The binding ratio is 1:2, and the binding constant is 2.4 × 10⁻⁶. 6 M -2 .

[0126] Example 27 Compound C10 in relation to Cu 2+ The infrared spectrum of the complex and its binding is shown in [reference]. Figure 7 .

[0127] Compound C10 and Cu 2+ After coordination, the characteristic peaks of the active sites of the hydroxyl group (O), heterocyclic N, and thiolactone carbonyl group (S) in the molecule all showed a significant red shift, and the characteristic absorption of the metal-ligand bond also appeared, fully demonstrating that C10 and Cu 2+ A stable coordination complex was successfully formed. Based on the molecular structural characteristics, Cu... 2+ The coordination mode is as follows: with the heterocyclic N atom and the thiolactone carbonyl S atom as the core coordination sites, and the hydroxyl O atom as the auxiliary coordination site, it interacts with Cu through multidentate chelation. 2+ A stable coordination structure is formed.

[0128] Example 28 Compound C10 in relation to Cu 2+ The spectra of the combined matter and its composite are shown in [reference]. Figure 8 .

[0129] The resulting complex C10-Cu 2+ ESI mass spectrometry characterization: [C10+2Cu] was detected in the spectrum. 2+ The molecular ion peak of / 2 is 252.4570 (HRMS calculation shows it to be 252.4578).

[0130] Example 29 Complex C10-Cu 2+ In vitro tumor proliferation inhibition activity test The in vitro activity test methods are as follows, and the results are shown in Table 2. Among them, paclitaxel, a commonly used anti-tumor drug in clinical practice, is the positive control experimental group.

[0131] (1) In vitro screening test for antitumor activity - 1 Screening method: microculture tetrozolium (MTT) reduction method; cell line: human lung cancer cells (A549); treatment time: 48 h; calculation of the C10-Cu complex. 2+ IC50 (half-maximal inhibitory concentration) for tumor cell growth 50 (%, μM).

[0132] (2) In vitro screening test for antitumor activity - 2 Screening method: microculture tetrozolium (MTT) reduction method; cell line: human colon cancer cells (HCT116); treatment time: 48 h; calculation of the C10-Cu complex. 2+ IC50 (half-maximal inhibitory concentration) for tumor cell growth 50 (%, μM).

[0133] (3) In vitro screening test for antitumor activity - 3 Screening method: microculture tetrozolium (MTT) reduction method; cell line: normal human kidney cells (HEK293T); treatment time: 48 h; calculation of the C10-Cu complex. 2+ IC50 (half-maximal inhibitory concentration) for tumor cell growth 50 (μM).

[0134] Table 2. C10-Cu complex 2+ In vitro tumor proliferation inhibitory activity

[0135] Example 30 Complex C10-Cu 2+ The XPS full spectrum and its fine copper element spectrum are shown in [reference needed]. Figure 9 .

[0136] XPS analysis revealed the complex C10-Cu 2+ The photoelectron spectrum of the C10-Cu complex. Based on the charge transfer phenomenon observed in the fluorescence titration spectroscopy experiment, the C10-Cu complex was analyzed. 2+ A fine scan of the copper peaks revealed that the spectrum showed a complex C10-Cu. 2+ It contains 2.63% copper. The complex is C10-Cu. 2+ The compound C10 contains monovalent copper, with a monovalent copper content of 45.5%, indicating that it can effectively reduce divalent copper to monovalent copper through charge transfer, thus enabling the C10-Cu complex to function as a monovalent copper compound. 2+Possesses stronger production capabilities OH ability.

[0137] Example 31 Complex C10-Cu 2+ Generation at different pH Performance tests of OH oxidation of TMB, results are shown in [link to results]. Figure 10 , 11 .

[0138] For the C10-Cu complex 2+ produce The properties of OH were tested. A colorless, transparent solution of 3,3',5,5'-tetramethylbenzidine (TMB) was subjected to... After oxidation with OH, a new UV absorption peak appears at 652 nm, and the solution turns blue. The solution turns blue at pH 5.0 and 6.8, but the change is not significant at pH 7.4. After adding sulfuric acid to stop the reaction, the solution turns yellow. (Compound C10-Cu) 2+ of The OH-generating activity was significantly higher than that of compound C10, confirming that... The efficient formation of OH does not originate from C10 itself, but rather from the reaction between C10 and Cu. 2+ The coordination complex formed is mediated.

[0139] Example 32 Complex C10-Cu 2+ GSH consumption test results are shown below. Figure 12 .

[0140] Complex C10-Cu 2+ The UV absorption at 412 nm shows a decreasing trend. At 1 h, the 20 mM C10-Cu complex... 2+ 70% of GSH was consumed at 1 h; after 3 h, 92% of GSH was consumed. The C10-Cu complex showed the following changes over time. 2+ The UV absorption peak at 412 nm decreases faster than that of compound C10. This indicates that compound C10 and Cu... 2+ It has a synergistic effect on consuming GSH.

[0141] Example 33 Complex C10-Cu 2+ Intracellular reactive oxygen species (ROS) detection results are shown in [Figure number missing]. Figure 13 , 14 .

[0142] Compound C10, Complex C10-Cu 2+After incubating A549 cells for 24 h, the cells were incubated for 20 min in a medium containing DCFH-DA dye. Fluorescence imaging showed that the negative control group cells exhibited very low green fluorescence intensity, while the cells incubated with compound C10 and the complex C10-Cu... 2+ Cells in the drug-treated group showed strong green fluorescence. This indicates the presence of compound C10 and the complex C10-Cu. 2+ It can enter A549 cells, and after administration, it promotes intracellular ROS generation and the formation of the C10-Cu complex. 2+ The green fluorescence intensity of the cells in the drug-treated group was significantly stronger than that of the cells in the compound C10-treated group.

[0143] In summary, in the C10-Cu complex 2+ It can enter A549 cells and promotes intracellular ROS generation after administration.

[0144] Example 34 Different concentrations of compound C10 and complex C10-Cu 2+ Analysis of mitochondrial membrane potential in A549 cells after drug administration; results are shown below. Figure 15 .

[0145] Compound C10 and the complex C10-Cu were detected by JC-1 staining. 2+ Changes in mitochondrial membrane potential in A549 cells after drug administration. We used HCT-116 cell lines containing compound C10 and the complex C10-Cu. 2+ The compounds were incubated for 12 h in complete culture medium at concentrations of 0 μM, 10 μM, 20 μM, and 50 μM, followed by JC-1 staining and fluorescence imaging. Compound C10 and the complex C10-Cu were also involved. 2+ The cells in all drug-treated groups showed similar fluorescence changes: at a drug concentration of 0 μM, the cells showed no green fluorescence but exhibited red fluorescence in the form of small mitochondrial particles; at a drug concentration of 10 μM, the cells showed both green and red fluorescence; at a drug concentration of 20 μM, the green fluorescence of the cells increased while the red fluorescence decreased; at a drug concentration of 50 μM, the cells emitted green fluorescence while the red fluorescence disappeared.

[0146] The results show that with the addition of compound C10 and the complex C10-Cu 2+ With increasing drug concentration, JC-1 in mitochondria changes from an aggregated form to a dispersed monomeric form, and the mitochondrial membrane potential changes from a normal state to an abnormal state. This will induce tumor cell apoptosis.

[0147] Example 35 Compound C10, Complex C10-Cu 2+ Apoptosis detection results are shown in [the table]. Figure 16 .

[0148] A549 cells were treated with the complex C10-Cu 2+ After incubation for 24 h, the complex was incubated for 10 min in 2'-(4-ethoxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5'-bis-1H-benzimidazole trihydrochloride trihydrate (Hoechst 33342 dye). After fluorescence imaging, the complex C10-Cu 2+ Cells in the drug-treated group exhibited indistinct cell membrane boundaries and irregular shapes; compared to the negative control group, Hoechst 33342 dye failed to stain the negative control group, and no blue fluorescence was observed; the C10-Cu complex... 2+ In the drug-treated group, cell nuclei exhibited blue fluorescence, and some cell nuclei showed bright white fluorescence. This phenomenon indicates that the C10-Cu complex... 2+ The drug can induce apoptosis in A549 cells.

Claims

1. A thiocamptothecin derivative, characterized in that: The thiocamptothecin derivatives have the following general structural formula: Wherein, R is selected from -H or -C(=O)-CH2-X-CH2-C(=O)-O-R1; X is selected from -S-, -O- or -CH2-; R1 is selected from C1-C6 alkyl or optionally benzyl substituted with one -CF3 or -CN.

2. The thiocamptothecin derivative according to claim 1, characterized in that: The thiocamptothecin derivative is one of the following compounds: 。 3. The method for preparing the thiocamptothecin derivative according to claim 1, characterized in that: (1) An alcohol compound R1-OH and an acid anhydride compound are dissolved in dichloromethane and esterified in the presence of a catalyst to obtain intermediate A. Wherein, the acid anhydride compound is selected from diethylene glycol anhydride, glutaric anhydride, or thioglycolic acid anhydride; R1 is selected from C1-C6 alkyl or optionally benzyl group substituted with one -CF3 or -CN; X is selected from -S-, -O-, or -CH2-. (2) Intermediate A is mixed with condensing agent and catalyst in dichloromethane to carry out an activation reaction to obtain an activated system; camptothecin is added to the activated system to carry out an esterification condensation reaction to obtain camptothecin derivative B. Wherein, R is the R described in claim 1; (3) Add an organic solvent to camptothecin derivative B or camptothecin, then add Lawson's reagent, and carry out a reflux reaction under nitrogen protection. Separate and purify to obtain the target thiocamptothecin derivative. or Wherein, R is the R described in claim 1; the organic solvent is xylene or dichloroethane.

4. The method for preparing thiocamptothecin derivatives according to claim 3, characterized in that: In step (1), the molar ratio of alcohol compound R1-OH to acid anhydride compound is 1.5:1; In step (2), the molar ratio of intermediate A to camptothecin is 3:1; In step (3), when the organic solvent is xylene, the molar ratio of camptothecin derivative B or camptothecin to Lawson reagent is 1:8~16; when the organic solvent is dichloroethane, the molar ratio of camptothecin derivative B or camptothecin to Lawson reagent is 1:

3.

5. The method for preparing thiocamptothecin derivatives according to claim 3, characterized in that: The catalyst is 4-dimethylaminopyridine; the condensing agent is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride.

6. The method for preparing thiocamptothecin derivatives according to claim 3, characterized in that: In step (1), the esterification reaction is carried out at room temperature. In step (2), the activation reaction condition is activation at room temperature for 30 min; the esterification condensation reaction condition is stirring at room temperature until the reaction is complete. In step (3), the heating and reflux reaction conditions are 80~130℃ for 1.5~3 h.

7. A complex based on thiocamptothecin derivatives, characterized in that: The complex is a combination of the thiocamptothecin derivative of claim 1 and Cu. 2+ Coordination complexes formed through multidentate chelation coordination.

8. The method for preparing the complex based on thiocamptothecin derivatives according to claim 7, characterized in that: Prepare equal volumes of solutions of the thiocamptothecin derivative of claim 1 and the divalent copper salt at a molar ratio of 1:2; mix the two solutions in equal volumes, stir thoroughly at room temperature, let stand, and allow to dry naturally to remove the solvent to obtain the final product.

9. The use of the thiocamptothecin derivative of claim 1 or the complex based on the thiocamptothecin derivative of claim 7 in the preparation of antitumor drugs.

10. The application according to claim 9, characterized in that: The tumor is one or both of lung cancer and colorectal cancer.

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