Preparation method and use of a sn-38-based prodrug cpt-1

Abstract

The application aims to disclose a preparation method and application of SN-38 prodrug CPT-1. Camptothecin (CPT) is an alkaloid with excellent antitumor activity isolated from plants of Camptotheca acuminata and Nothapodytes pittosporoides. Among various camptothecin derivatives, 7-ethyl-10-hydroxycamptothecin (SN-38) has very good biological activity. In the application, SN-38 is used as a parent molecule for structural derivation, and SN-38 is coupled with benzisoselenazolone (NBD-Se) to obtain a novel prodrug molecule CPT-1. Studies show that CPT-1 can release drug molecules SN-38 and benzisoselenazolone with photosensitive effect in situ after reacting with glutathione (GSH), so as to achieve the purpose of synergistic antitumor.

Description

Technical Field

[0001] This invention relates to the field of chemical pharmaceuticals, and more specifically, to a method for preparing the SN-38-based prodrug CPT-1 and its uses. Background Technology

[0002] Cancer is one of the major diseases that seriously threaten human health and life, with complex pathogenic factors and high incidence and mortality rates. Chemotherapy remains one of the main methods for the clinical treatment of malignant tumors. Conventional chemotherapy drugs mostly suffer from poor targeting, significant toxic side effects, low bioavailability, and drug resistance, resulting in poor clinical efficacy and poor quality of life for patients. Camptothecin (CPT) is an alkaloid with excellent antitumor activity isolated from the plants Camptothecinus and *Cephalotaxus fortunei*. Its chemical structure contains the lactone ring (E ring), pyridone ring (D ring), and 20-hydroxyl group, which are essential groups for camptothecin to exert its topoisomerase I inhibitory effect, thereby achieving its antitumor effect. Appropriate modification of camptothecin at C7, C9, and C10 often enhances its antitumor effect. Among various camptothecin derivatives, 7-ethyl-10-hydroxycamptothecin (SN-38) possesses very good biological activity. Activation of molecular diagnostic reagents generally involves breaking chemical bonds in the tumor microenvironment, which differs from that of normal tissue, releasing the original drug molecules and fluorescent imaging molecules to achieve diagnostic and therapeutic effects. The tumor microenvironment is characterized by high concentrations of biological sulfhydryl groups, high levels of reactive oxygen species, low pH, and unique enzyme activities. Therefore, stimulation-responsive drugs induced by the tumor microenvironment can significantly enhance the efficacy of anticancer drugs by increasing drug concentration at the tumor site. Based on this, this invention, for the first time, discloses a novel SN-38-based prodrug activated by glutathione (GSH) using benzo[selenium diazole] (NBD-Se), a photosensitive compound, as a recognition site, to achieve synergistic targeted cancer therapy. Summary of the Invention

[0003] Based on the current state of existing technology research, this invention provides a method for preparing the SN-38-based prodrug CPT-1 and its applications. The CPT-1 described in this invention can exert a synergistic effect, thereby killing tumor cells. Attached Figure Description

[0004] Figure 1 This is a schematic diagram showing the cytotoxic activity results of CPT-1 and SN-38 on HeLa cells in Example 2.

[0005] Figure 2 This is the high-performance liquid chromatogram of CPT-1 reacted with GSH in Example 3.

[0006] Figure 3This is a graph showing the release efficiency of SN-38 after the interaction of CPT-1 and GSH in Example 3.

[0007] Example 1

[0008] This invention discloses a prodrug molecule CPT-1 based on SN-38, the structure of which is shown in the following formula: ; The synthetic route of CPT-1 described in this invention is as follows:

[0009] The specific preparation method of CPT-1 described in this invention is as follows: Synthesis of compound CPT-1: SN-38 (78 mg, 0.2 mmol) was dissolved in acetonitrile solution, followed by the addition of K₂CO₃ (56 mg, 0.4 mmol) and NBD-Se (53 mg, 0.2 mmol). The mixture was refluxed and stirred until the reactants were completely reacted. The reaction progress was monitored by TLC. After the reaction was completed, the acetonitrile solution was removed by rotary evaporation, and the final target compound CPT-1 was obtained by column chromatography. Specific structural identification data are as follows: 1 H NMR (400 MHz, CDCl3) δ 8.16 (dd, J = 9.1, 5.5 Hz, 2H), 7.70 (d, J = 6.2 Hz, 2H), 7.40 (d, J = 9.3 Hz, 2H), 7.22 (s, 1H), 5.30 (s, 4H), 3.49 (s, 2H), 0.64 (s, 3H); 13 C NMR (100 MHz, CDCl3) δ 172.5,156.9,154.6,153.3,149.9,146.7, 142.2, 139.5,137.5, 136.8, 133.9,132.7, 130.0,126.0,121.4,119.9,113.6,104.8,100.7,74.4,65.7,56.5,30.3,24.7, 15.3, 7.7.

[0010] Example 2: Cytotoxic activity assay of prodrug molecules CPT-1 and SN-38 against HeLa cells.

[0011] Experimental methods: 5000 HeLa cells were added to each well of a 96-well plate and treated with CPT-1 and SN-38 for 24 h. The concentrations of CPT-1 and SN-38 were 0 uM, 2 uM, 5 uM, 10 uM, 20 uM and 50 uM, respectively. Finally, the cytotoxicity of CPT-1 and SN-38 was determined by the MTT assay.

[0012] Experimental results: The results are as follows Figure 1 As shown, the results indicate that the prodrug molecule CPT-1 has less cytotoxicity than the drug molecule SN-38, and that the prodrug molecule CPT-1 has the ability to release the drug slowly.

[0013] Example 3: Investigation into the mechanism of action of the prodrug molecule CPT-1.

[0014] Experimental method: CPT-1 (10 uM) and GSH (100 uM) were incubated in DMSO / PBS (50:50, v / v) for a specific time before being injected for testing.

[0015] Experimental results: such as Figure 2 and Figure 3 As shown, after CPT-1 reacts with GSH, the amount of SN-38 drug molecule released gradually increases with increasing reaction time, reaching equilibrium after 4 hours. Therefore, it can be concluded that after 4 hours of reaction between CPT-1 and GSH, SN-38 is released, with a maximum drug release rate of 68%.

Claims

1. An anticancer prodrug molecule CPT-1, having the structural formula: 。 2. Use of CPT-1 as claimed in claim 1, characterized in that The CPT-1 and the CPT-2 can play a synergistic role, and then kill tumor cells.

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