Preparation method and application of prodrug CPT-1 based on SN-38

Abstract

The invention aims at disclosing a preparation method and application of an SN-38 prodrug CPT-1. Camptothecin (CPT) is an alkaloid which is separated from plant camptotheca acuminata and Camptotheca nodiflora and has excellent anti-tumor activity, and 7-ethyl-10-hydroxycamptothecin (SN-38) in various camptothecin derivatives at present has very good biological activity. According to the present invention, SN-38 is adopted as a parent molecule to carry out structure derivation, and SN-38 and benzoselenadiazole (NBD-Se) are coupled to obtain the novel prodrug molecule CPT-1; researches show that after the CPT-1 acts with glutathione (GSH), a drug molecule SN-38 and benzoselenadiazole with a photosensitive effect can be released in situ, and the purpose of synergistic anti-tumor is achieved.

Description

Technical Field

[0001] The present invention relates to the field of chemical medicine, and in particular to a preparation method and use of a SN-38-based prodrug CPT-1. Background Art

[0002] Cancer is one of the major diseases that seriously threaten human health and life. Its pathogenic factors are complex, and its morbidity and mortality are high. Chemotherapy is still one of the main means of clinical treatment of malignant tumors. Most conventional chemotherapy drugs have the defects of poor targeting, large toxic side effects, low bioavailability and drug resistance, which leads to poor clinical efficacy and poor quality of life of patients. Camptothecin (CPT) is an alkaloid with excellent antitumor activity isolated from the plants Camptotheca acuminata and Camptotheca sempervirens. The lactone ring (E ring), pyridone ring (D ring) and 20-hydroxyl in its chemical structure are essential groups for camptothecin to exert topoisomerase I inhibition and achieve antitumor effect. The antitumor effect of camptothecin can often be enhanced by appropriate modification of C7, C9 and C10. Among various camptothecin derivatives, 7-ethyl-10-hydroxycamptothecin (SN-38) has very good biological activity. The activation of molecular diagnostic and therapeutic agents is generally achieved by breaking chemical bonds between the tumor tissue microenvironment and normal tissue, releasing the original drug molecules and fluorescent imaging molecules, so as to achieve the purpose of diagnostic and therapeutic effects. The tumor microenvironment mainly includes high concentration of biological thiol groups, high level of reactive oxygen species, low pH value, unique enzyme activity, etc. Therefore, stimulus-responsive drugs induced by the tumor microenvironment can significantly enhance the efficacy of anticancer drugs by increasing the drug concentration at the tumor site. Based on this, the present invention uses benzoselenadiazole (NBD-Se) with photosensitivity as a recognition site for the first time, and discloses a new type of diagnostic and therapeutic prodrug based on SN-38 activated by glutathione (GSH) to achieve the purpose of synergistic targeted cancer treatment. Summary of the invention

[0003] Based on the current status of the above-mentioned prior art research, the present invention provides a preparation method and use of a SN-38-based prodrug CPT-1. The CPT-1 of the present invention can exert a synergistic effect and thereby kill tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0005] Figure 2 This is the HPLC chromatogram after the CPT-1 in Example 1 reacted with GSH.

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

[0007] Example 1

[0008] The present invention discloses a prodrug molecule CPT-1 based on SN-38, the structure of which is shown in the following formula:

[0009]

[0010] The synthetic route of CPT-1 described in the present invention is as follows:

[0011]

[0012] The specific preparation method of CPT-1 of the present invention is as follows:

[0013] Synthesis of compound CPT-1: SN-38 (78 mg, 0.2 mmol) was dissolved in acetonitrile solution, and then K 2 CO 3 (56 mg, 0.4 mmol) and NBD-Se (53 mg, 0.2 mmol), reflux and stir until the raw materials are completely reacted, the reaction progress is detected by TLC, the reaction is completed, the acetonitrile solution is removed by rotary evaporation, and the final target compound CPT-1 is obtained by column chromatography separation. The specific structural identification data are as follows: 1 H NMR (400 MHz, CDCl 3 )δ8.16(dd,J=9.1,5.5Hz,2H),7.70(d,J=6.2Hz,2H),7.40(d,J=9.3Hz,2H),7.22(s,1H),5.30(s,4H),3.49(s,2H),0.64(s,3H); 13 C NMR (100 MHz, CDCl 3 )δ172.5,156.9,154.6,153.3,149.9,146.7,142.2,139.5,137.5,136.8,133.9,132.7,130 .0,126.0,121.4,119.9,113.6,104.8,100.7,74.4,65.7,56.5,30.3,24.7,15.3,7.7,0.00.

[0014] Example 1: Cytotoxic activity test of prodrug molecules CPT-1 and SN-38 on HeLa cells.

[0015] Experimental method: 5000 HeLa cells were added to each well of a 96-well plate, and the cells were treated with CPT-1 and SN-38 for 24 hours. The concentrations of CPT-1 and SN-38 were divided into 0uM, 2uM, 5uM, 10uM, 20uM, and 50uM, respectively. Finally, the cytotoxicity of CPT-1 and SN-38 was determined by the MTT method.

[0016] Experimental results: The results are as follows Figure 1 As shown, the results indicate that the cytotoxicity of the prodrug molecule CPT-1 is lower than that of the drug molecule SN-38, and the prodrug molecule CPT-1 has the ability to slowly release drugs.

[0017] Example 2: Exploration of the mechanism of action of the prodrug molecule CPT-1.

[0018] Experimental method: CPT-1 (10uM) and GSH (100uM) were incubated in DMSO / PBS (50:50, v / v) for a specific period of time and then injected for testing.

[0019] Experimental results: Figure 2 and Figure 3 As shown in the figure, after CPT-1 reacts with GSH, the amount of drug molecule SN-38 released gradually increases with the increase of reaction time, and the reaction reaches equilibrium after the response time is 4 hours. Therefore, it can be concluded that after CPT-1 reacts with GSH for 4 hours, SN-38 is released, and the maximum drug release rate is 68%.

Claims

1. The present invention discloses a preparation method and use of a SN-38 analogue CPT-1. The structure of the CPT-1 of the present invention is shown in the following formula (I). The CPT-1 of the present invention can play a synergistic role and kill tumor cells; 2. CPT-1, characterized in that Its structure is shown in the following formula (I):

3. A use of CPT-1 as claimed in claim 1 or 2, characterized in that They can play a synergistic role and kill tumor cells.

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