CN119074718A

CN119074718A - Application of dihydroartemisinin combined with chrysin in the preparation of products for improving the sensitivity of tumor cells to TOP2A inhibitors

Info

Publication number: CN119074718A
Application number: CN202411210976.1A
Authority: CN
Inventors: 薛洪省; 孙海迪; 郎明非; 孙晶; 韩丽; 张屹
Original assignee: Dalian University
Current assignee: Dalian University
Priority date: 2024-08-30
Filing date: 2024-08-30
Publication date: 2024-12-06

Abstract

本发明属于生物医药技术领域，公开了双氢青蒿素联合白杨素在制备提高肿瘤细胞对TOP2A抑制剂敏感性的产品中的应用。其中TOP2A抑制剂为依托泊苷，具体用于阻碍DNA修复。本发明为现有TOP2A抑制剂耐药提出了新的解决方案，充分利用并延长现有药物的生命周期从医药经济学原则上说更符合目前的医药工业水平和医保投资现状。The present invention belongs to the field of biomedicine technology, and discloses the use of dihydroartemisinin combined with chrysin in the preparation of a product that increases the sensitivity of tumor cells to TOP2A inhibitors. The TOP2A inhibitor is etoposide, which is specifically used to hinder DNA repair. The present invention proposes a new solution for the existing TOP2A inhibitor resistance, fully utilizing and extending the life cycle of existing drugs, which is more in line with the current level of the pharmaceutical industry and the current status of medical insurance investment from the perspective of pharmaceutical economics.

Description

Application of dihydroartemisinin combined with chrysin in preparation of product for improving sensitivity of tumor cells to TOP2A inhibitor

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to application of dihydroartemisinin combined with chrysin in preparation of a product for improving sensitivity of tumor cells to TOP2A inhibitors.

Background

Lung cancer, one of the common malignant tumors that threatens human life, has extremely high morbidity and mortality, and is classified into non-small cell molecules (NSCLC) and Small Cell Lung Cancer (SCLC). The lung cancer is mainly treated by surgery, radiotherapy and chemotherapy, immunotherapy and targeted drug therapy. However, the chemotherapy drugs widely used in clinic still have serious toxic and side effects, such as nausea and vomiting, bone marrow transplantation, heart toxicity, neurotoxicity and the like, which may cause the situation of patients to be worsened, and the problems to be solved of cancer are tumor heterogeneity and drug resistance, so that the research on exploring novel anti-cancer drugs with low toxicity and effectiveness is the focus of attention of domestic and foreign scientists, and the research discovers that the combination therapy of different drugs can partially improve the problems.

Etoposide (Etoposide, VP-16), having the chemical formula C29H32O13, is an effective topoisomerase II inhibitor and can inhibit DHA repair. DNA damage repair is an important mechanism of drug resistance, one of the main causes of metastasis and recurrence after chemotherapy. Etoposide has been widely used clinically as a chemotherapeutic agent incorporated into the "clinical diagnosis and treatment guidelines for lung cancer of the chinese medical society", and has serious toxic and side effects, including myelosuppression, mucositis, cardiotoxicity, nephrotoxicity, alopecia and immunosuppression, as with other chemotherapeutic agents.

Dihydroartemisinin (Dihydroartemisinin; DHA) is an active metabolite of artemisinin and its derivatives (ARTs), is an effective clinical drug widely used for the treatment of malaria, and DHA has been found to show potential anti-tumor effects in cancer treatment. Studies show that DHA plays an anticancer role through a plurality of molecular mechanisms such as apoptosis induction, proliferation inhibition, tumor metastasis inhibition, angiogenesis inhibition, immunity promotion, autophagy induction, endoplasmic Reticulum (ER) stress induction and the like. Meanwhile, researches have proved that DHA reduces side effects caused by cisplatin treatment and reverses drug resistance of cisplatin. Notably, DHA has synergistic anti-tumor effect with various clinical drugs.

Chrysin (Chrysin; CRS) is a natural flavonoid compound which is present in honey and various plant extracts and has wide pharmacological and physiological activities, and chrysin is considered as a promising drug for treating diseases in view of its wide distribution and relatively low toxicity. In recent years chrysin has been attracting attention for its anti-tumor and antioxidant activity and its protective effect against allergic inflammation. Chrysin administration can improve lipid peroxidation and antioxidant status, thereby modulating oxidant/antioxidant balance in cancer. The main anticancer mechanism of chrysin includes inhibition of cell proliferation, inhibition of inflammation and induction of apoptosis. In addition, the chrysin can be used in combination with clinical chemotherapy drug pyrroltinib to play the role of resisting breast cancer.

Topoisomerase II (TOP 2) is a nuclear protein required for DNA replication and cell division. Thus, TOP2 is the primary cellular target for many of the most widely used potent chemotherapeutics, including doxorubicin, etoposide, and mitoxantrone. Topoisomerase II has two different gene isoforms (TOP 2A and TOP 2B), which are expressed and regulated differently in living cells. TOP2B is expressed in resting cells of almost all tissues throughout the cell cycle. Whereas TOP2A is typically expressed at high levels in rapidly proliferating and growing cells. DNA topoisomerase II (TOP 2A) is closely related to the occurrence, invasion, treatment and prognosis of malignant tumors. It regulates the biological behavior of tumors by participating in various biological processes such as cell cycle and apoptosis. And is closely related to the target point of tumor chemotherapeutic drugs.

The traditional Chinese medicine has the advantages of large yield, small toxic and side effects, multiple targets and the like. In lung cancer treatment, traditional Chinese medicines have been proved to be capable of prolonging survival rate and have less toxic and side effects. A large number of researches show that the traditional Chinese medicine can not only relieve clinical symptoms of cancer patients and improve the quality of life of the cancer patients, but also improve the effects of chemotherapy and radiotherapy and reduce adverse reactions and complications caused by radiotherapy or targeted therapy. With the deep treatment of cancer, the drug resistance of patients is gradually enhanced, and the traditional Chinese medicine combined chemotherapy plays a synergistic role in improving the chemotherapy effect and reducing the toxic and side effects.

Disclosure of Invention

In order to overcome the defects in the prior art, the invention provides application of dihydroartemisinin combined with chrysin in preparation of products for improving sensitivity of tumor cells to TOP2A inhibitors.

The aim of the invention is realized by the following technical scheme that the dihydroartemisinin and chrysin are combined to prepare the product for improving the sensitivity of tumor cells to TOP2A inhibitor.

Furthermore, the application of dihydroartemisinin combined with chrysin in preparing products for improving the sensitivity of tumor cells to TOP2A inhibitors, and TOP2A inhibitors are particularly used for preventing DNA repair.

Further, the TOP2A inhibitor is etoposide.

Further, the concentration ratio of dihydroartemisinin to chrysin is dihydroartemisinin to chrysin=20. Mu.M to 25. Mu.M or 11. Mu.M to 30. Mu.M.

Further, the tumor is lung cancer.

Further preferred of the present invention, the lung cancer is small cell lung cancer and non-small cell lung cancer.

Another object of the invention is to protect a pharmaceutical composition for increasing the sensitivity of tumor cells to TOP2A inhibitors, comprising TOP2A inhibitors, dihydroartemisinin and chrysin.

Further, TOP2A inhibitor in the pharmaceutical composition is etoposide, and dihydroartemisinin is 20 mu M, 25 mu M, 5.22 mu M or 11 mu M, 30 mu M and 5.221 mu M of etoposide.

Further, the pharmaceutical composition also comprises pharmaceutically acceptable auxiliary materials.

Compared with the prior art, the invention has the beneficial effects that the research discovers that the dihydroartemisinin is combined with the chrysin and the TOP2A inhibitor for use, so that the sensitivity of the TOP2A inhibitor can be increased, the dosage can be reduced, and the treatment time can be prolonged. The pharmaceutical composition of the invention inhibits TOP2A expression based on dihydroartemisinin combined with chrysin, causes DNA damage, promotes lung cancer cell apoptosis, thereby improving the sensitivity of lung cancer cells to TOP2A inhibitor, enhancing the therapeutic effect of the drug, and can be used for treating the occurrence of TOP2A inhibitor drug resistance. Can be used for treating TOP2A inhibitor drug resistance. The invention provides a new solution for the drug resistance of the existing TOP2A inhibitor, fully utilizes and prolongs the life cycle of the existing drug, and accords with the current medical industry level and medical insurance investment state in principle of medicine economy.

Drawings

The invention will be further described with reference to the drawings and the detailed description

Fig. 1 is a graph showing the effect of dual artemisinin in combination with chrysin in inhibiting the activity of etoposide in lung cancer cells in different lung cancer cell lines (x indicates that the concentration of the dual artemisinin is 20 μm/11 μm and the final concentration of chrysin is 25 μm/30 μm compared to the DHA half-to-dead concentration treatment group; x indicates that the concentration of the dual artemisinin is 20 μm/11 μm and the final concentration of chrysin is 5.22 μm (#/< 0.05, #/# P <0.01, # # #/# P <0.001, # #/# P <0.0001, n=3);

Fig. 2 is a graph showing the effect of combination drug and etoposide, dihydroartemisinin in combination with chrysin on normal epithelial cell activity of human lung (< 0.05P <0.01, P <0.001, P <0.0001, n=3);

Fig. 3 is a graph showing the effect of combination drugs on TOP2A expression and DNA damage (< 0.05, P <0.01, P <0.001, P <0.0001, n=3);

fig. 4 is a graph showing the effect of combination drugs on apoptosis (< 0.05, P <0.01, P <0.001, P <0.0001, n=3).

Detailed Description

The present invention is described in detail below by way of specific examples, but the scope of the present invention is not limited thereto. Unless otherwise specified, the experimental methods used in the present invention are all conventional methods, and all experimental equipment, materials, reagents, etc. used can be obtained from commercial sources.

Materials and instruments

Example 1

A pharmaceutical composition for increasing the sensitivity of tumor cells to TOP2A inhibitors, said medicament comprising dihydroartemisinin, chrysin and TOP2A inhibitors, the final concentrations of TOP2A inhibitors, dihydroartemisinin, chrysin and the type of antibiotics in the pharmaceutical composition are shown in table 1.

TABLE 1

The preparation method of the pharmaceutical composition of the embodiment comprises the following steps:

(1) 11.77mg of etoposide is weighed by a ten-thousandth scale and dissolved in 2ml of DMSO solvent to prepare a 10mM stock solution, 142.18mg of dihydroartemisinin is weighed and dissolved in 5ml of DMSO solvent to prepare a 100mM stock solution, and 127.12mg of chrysin is weighed and dissolved in 5ml of DMSO solvent to prepare a 100mM stock solution.

(2) Stock solution of etoposide was diluted to 20.88. Mu.M using 1640 medium containing 10% serum, stock solution of dihydroartemisinin was diluted to 20. Mu.M and 11. Mu.M, and stock solution of chrysin was diluted to 25. Mu.M and 30. Mu.M.

(3) The stock solution of etoposide was diluted to 20.88. Mu.M using a DMEM low-sugar medium containing 10% serum, the stock solution of dihydroartemisinin was diluted to 20. Mu.M and 11. Mu.M, and the stock solution of chrysin was diluted to 25. Mu.M and 30. Mu.M.

(4) Etoposide, dihydroartemisinin, chrysin, 10.44. Mu.M, 20. Mu.M, 25. Mu.M/5.22. Mu.M, 20. Mu.M/25. Mu.M/2.61. Mu.M, 20. Mu.M, 25. Mu.M working fluid was prepared using 1640 medium with 10% serum.

(5) A1640 medium containing 10% serum was used to prepare working solution of etoposide, dihydroartemisinin, chrysin, 10.44. Mu.M, 11. Mu.M, 30. Mu.M/5.22. Mu.M, 11. Mu.M, 30. Mu.M/2.61. Mu.M, 11. Mu.M, 30. Mu.M.

(6) A working solution of etoposide, dihydroartemisinin, chrysin, 10.44. Mu.M, 20. Mu.M, 25. Mu.M/5.22. Mu.M, 20. Mu.M, 25. Mu.M/2.61. Mu.M, 20. Mu.M, 25. Mu.M was prepared using a DMEM low-sugar medium containing 10% serum.

(7) A working solution of etoposide, dihydroartemisinin, chrysin, 10.44. Mu.M, 11. Mu.M, 30. Mu.M/5.22. Mu.M, 11. Mu.M, 30. Mu.M/2.61. Mu.M, 11. Mu.M, 30. Mu.M was prepared using a DMEM low-sugar medium containing 10% serum.

(8) A DMEM high-sugar culture medium containing 10% serum was used to prepare 5.22. Mu.M working solution of etoposide, 20. Mu.M of dihydroartemisinin, 25. Mu.M/11. Mu.M of chrysin, 30. Mu.M working solution of dihydroartemisinin, 5.22. Mu.M of dihydroartemisinin, 20. Mu.M/25. Mu.M/5.22. Mu.M, 11. Mu.M and 30. Mu.M of chrysin.

Example 2

Etoposide alone, dihydroartemisinin combined with chrysin, and effect of etoposide, dihydroartemisinin and chrysin combined medicines on different cell activities

The method comprises inoculating lung cancer cells H446 into 96-well plate with the amount of about 6.5X10- ⁴ cells/well, inoculating for 14 hr, using etoposide 20.88 μM, dihydroartemisinin 20 μM 25 μM, etoposide 20.44 μM dihydroartemisinin 25 μM/5.22 μM 20 μM 25 μM/2.61 μM 20 μM 25 μM treated cells, transferring to 37 ℃ constant temperature CO ₂ incubator, incubating for 1 hr with CCK837 ℃ constant temperature CO ₂ incubator, and detecting absorbance with enzyme marker instrument. The absorbance values reflect cell viability. The blank group was set without any drug intervention.

The method comprises inoculating lung cancer cells A549 into 96-well plate with the amount of about 1.0X10- ⁵ cells/well, inoculating for 14h, using etoposide 20.88 μM, dihydroartemisinin 11 μM 30 μM, etoposide 11.44 μM 11 μM 30 μM/5.22 μM 11 μM 30 μM/2.61 μM 11 μM 30 μM treated cells, transferring to 37 ℃ constant temperature CO ₂ incubator for overnight treatment, adding CCK837 ℃ constant temperature CO ₂ incubator for incubation for 1h, and detecting absorbance by using enzyme marker instrument. The absorbance values reflect cell viability. The blank group was set without any drug intervention.

The method comprises inoculating normal epithelial cells BEAS-2B of human lung into 96-well plate with the number of 1.0X10- ⁵/well, inoculating for 14 hr, incubating with etoposide 20.88 μm, dihydroartemisinin 20. Mu.M 25. Mu.M/11. Mu.M 30. Mu.M, etoposide 5.22. Mu.M 20. Mu.M/5.22. Mu.M 11. Mu.M 30. Mu.M, transferring to 37 deg.C constant temperature CO ₂ incubator overnight, incubating with CCK837 deg.C constant temperature CO ₂ incubator for 1 hr, and detecting absorbance with enzyme-labeled instrument. The absorbance values reflect cell viability. The blank group was set without any drug intervention.

TABLE 2 Dihydroartemisinin in combination with chrysin and etoposide alone treatment of H446 and A54924H before IC ₅₀ concentration is reached

Drug treatment	IC ₅₀ are respectively
		Dihydroartemisinin-chrysin	20μM:25μM
Dihydroartemisinin-chrysin	11μM:30μM
		Etoposide	20.88μM

TABLE 3 influence of etoposide, dihydroartemisinin and chrysin combination on cell viability after H44624H treatment

Drug treatment	Drug concentration	Cell viability
			Etoposide	20.88μM	54.18%
Dihydroartemisinin-chrysin	20μM:25μM	53.06%
			Etoposide, dihydroartemisinin and chrysin	10.44μM:20μM:25μM	38.42%
Etoposide, dihydroartemisinin and chrysin	5.22μM:20μM:25μM	53.46%
			Etoposide, dihydroartemisinin and chrysin	2.61μM:20μM:25μM	68.45%

TABLE 4 influence of etoposide, dihydroartemisinin and chrysin combination treatment A54924h on cell viability

Drug treatment	Drug concentration	Cell viability
			Etoposide	20.88μM	53.34%
Dihydroartemisinin-chrysin	11μM:30μM	53.78%
			Etoposide, dihydroartemisinin and chrysin	10.44μM:11μM:30μM	42.11%
Etoposide, dihydroartemisinin and chrysin	5.22μM:11μM:30μM	52.78%
			Etoposide, dihydroartemisinin and chrysin	2.61μM:11μM:30μM	68.16%

TABLE 5 Effect of different drug combination concentrations on cell viability after 24h treatment of human lung normal epithelial cells BEAS-2B

Drug treatment	Drug concentration	Cell viability
			Etoposide	20.88μM
Etoposide, dihydroartemisinin and chrysin	5.22μM:11μM:30μM
			Etoposide, dihydroartemisinin and chrysin	5.22μM:11μM:30μM

Experimental results

The CCK8 experiment is used for detecting the cell viability, the experimental results are shown in figures 1,2, 3, 4 and 5, compared with the combined medicament of etoposide, double artemisinin and chrysin, the combined medicament of etoposide, double artemisinin and chrysin has the same inhibiting effect on the survival rate of lung cancer cells H446 and A549, the influence on the survival rate of normal lung epithelial bronchial cells BEAS-2B is obviously reduced, and under the condition of reaching IC ₅₀, the dosage of the combined medicament is lower than that of the single medicament, and the safety of the combined medicament is higher.

Example 3

The combination of dual artemisinin and chrysin increases DNA damage to lung cancer cells.

The implementation method comprises inoculating lung cancer cells H446 and A549 into a 24-well plate, inoculating for 14H, preparing etoposide, dihydroartemisinin, chrysin, 5.22 mu M, 20 mu M, 25 mu M (H446 cells)/5.22 mu M, 11 mu M, 30 mu M (A549 cells), treating the cells respectively, adding 1640 culture medium with 10% serum or DMEM culture medium with 10% serum into a control group, transferring into a cell culture box after the treatment, and taking out after 24H. 250 μ L RNAKEY REAGENT was added per well, and the lysed cell fluid was blown and collected into a 1.5mL Ep tube. A volume of 0.1mL of chloroform instead of BCP was added per 1mL RNAkey Reagent, shaken for 15s, allowed to stand at room temperature for 3 minutes, and centrifuged at 12000g for 10 minutes at 4 ℃. The upper aqueous phase was transferred to a fresh Ep tube, 0.5mL of isopropanol was added, allowed to stand at room temperature for 10 minutes, centrifuged at 12000g for 10 minutes at 4 ℃, the supernatant was discarded, the bottom white precipitate was retained, 1mL of 75% ethanol was added, the white precipitate was washed by blowing, centrifuged at 12000g for 10 minutes at 4 ℃, the waste liquid was discarded, the cover was opened and back-buckled on alcohol paper for 10 minutes, 50 mu LRNA of the solution was added, and the mixture was water-bath at 55℃for 10 minutes. Finally, the concentration and quality of RNA are measured by an enzyme-labeled instrument (the quality reference OD _260/280 is between 1.8 and 2.0). The RNA was reverse transcribed to obtain cDNA, which was subsequently used in qPCR experiments.

The method comprises spreading the cell slide in 24-well plate, inoculating lung cancer cells H446 and A549 in 24-well plate containing cell slide, inoculating for 14 hr, adding 4% paraformaldehyde fixed solution (150 μL per well plate) of etoposide (dihydroartemisinin: chrysin: 5.22 μM:20 μM (H446 cells)/5.22 μM:11 μM:30 μM (A549 cells), treating the cells respectively, adding 1640 medium with 10% serum or DMEM medium with 10% serum to control group, transferring to cell incubator after treatment, taking out after 24 hr, adding 150 μL per well at room temperature (150 μL per well plate for each time under no special explanation), washing with PBS for 20 min, permeabilizing cell membrane with PBS for 10 min, washing with PBS for three times for 5min, adding serum for 1 hr, and incubating at 200 ℃ for 1 hr under 4 ℃ in refrigerator, and sealing the antibody concentration of TOP after 200 deg.C overnight. The following day the 24 well plate was removed, washed three times with PBS for 5 minutes each, with Dylight 488,GoatAnti Mouse IgG (dilution 1:500) added, incubated for 1 hour at room temperature, three times with PBS for 5 minutes each, stained with 0.6. Mu.g/mL DAPI, washed three times with PBS for 5 minutes after 10 minutes of action, the cell slide in the 24 well plate was removed and inverted onto a slide with anti-fluorescence quencher dropped, and finally the slide was transferred to immunofluorescence microscopy in dark environment for immunofluorescence photography.

The method comprises spreading the cell slide in 24-well plate, inoculating lung cancer cells H446 and A549 in 24-well plate containing cell slide, inoculating for 14 hr, adding 4% paraformaldehyde fixed solution (150 μL per well plate) of etoposide (dihydroartemisinin: chrysin: 5.22 μM (H446 cells)/5.22 μM:11 μM:30 μM (A549 cells), treating the cells respectively, adding 1640 medium with 10% serum or DMEM medium with 10% serum to control group, transferring to cell incubator after treatment, taking out after 24 hr, adding 150 μL per well at room temperature (150 μL per well plate for each time under no special explanation), washing with PBS for 20 min, adding 0.3% Triton X-100 to permeabilize cell membrane for 10 min, washing with PBS for three times, transferring to goat antibody after adding serum to treat cell for 1 hr, and sealing with antibody at 200 deg.C after 1 deg.C overnight at refrigerator. The following day the 24 well plate was removed, washed three times with PBS for 5 minutes each, with Dylight 488,GoatAnti Mouse IgG (dilution 1:500) added, incubated for 1 hour at room temperature, three times with PBS for 5 minutes each, stained with 0.6. Mu.g/mL DAPI, washed three times with PBS for 5 minutes after 10 minutes of action, the cell slide in the 24 well plate was removed and inverted onto a slide with anti-fluorescence quencher dropped, and finally the slide was transferred to immunofluorescence microscopy in dark environment for immunofluorescence photography.

Experimental results

The experiment can further inhibit TOP2A expression at mRNA level and protein level and further increase DNA damage occurrence after combined treatment of etoposide, dihydroartemisinin and chrysin combined medicines through immunofluorescence experiment detection. This demonstrates that the combined use of the three drugs exerts a synergistic effect in inhibiting TOP2A expression and increasing DNA damage.

Example 4

The apoptosis of lung cancer cells is caused by the combined drug action of etoposide, dihydroartemisinin and chrysin.

The implementation method comprises the steps of paving a cell slide in a 24-well plate, inoculating lung cancer cells H446 and A549 in the 24-well plate containing the cell slide, inoculating for 14 hours, then, taking out etoposide, dihydroartemisinin, chrysin, 5.22 mu M, 20 mu M, 25 mu M (H446 cells)/5.22 mu M, 11 mu M, 30 mu M (A549 cells), respectively treating the cells, adding 1640 culture medium with 10% serum or DMEM culture medium with 10% serum into a control group, transferring the treated cells into a cell culture box, taking out the cell culture box after the treatment is finished, discarding waste liquid, adding 5 mu lAnnexinV-FITC, 5 mu L PI STAINING Solution and 140 mu L of corresponding culture medium, incubating for 10 minutes at room temperature in a dark place, taking out the cell slide in the 24-well plate, inverting the cell slide with the anti-fluorescence quencher, and finally transferring the slide to a dark environment for immunofluorescence photographing under a immunofluorescence microscope.

The method comprises inoculating lung cancer cells H446 and A549 to 6-well plate, inoculating for 14 hr, preparing etoposide, dihydroartemisinin, chrysin, 5.22 μM, 20 μM, 25 μM (H446 cells)/5.22 μM, 11 μM, 30 μM (A549 cells), treating the cells respectively, adding 1640 culture medium containing 10% serum or DMEM culture medium containing 10% serum to control group, transferring to cell incubator after treatment, taking out after 24 hr, digesting the cells with pancreatin without EDTA, centrifuging at 1800rpm and 4 ℃ for 5 min, discarding supernatant, washing the cells twice with PBS, centrifuging at 1800rpm and 4 ℃ for 5 min each time, discarding supernatant, adding 100 μL of 1×binding Buffer, gently blowing to single cell suspension, adding 5 μL of lAnnexinV-FITC and 5 μL L PI STAINING Solution, gently blowing, incubating at room temperature for 10 min, adding 400 μL of 1×binding Buffer, gently mixing, and transferring to single-dye to a filter for single-dye in PI flow meter for detecting apoptosis, and filtering, and testing.

Experimental results

The experimental detection result of the flow cytometry shows that the combined treatment of etoposide, dihydroartemisinin and chrysin causes the increase of lung cancer cell apoptosis. This demonstrates that the combined use of the three drugs exerts a synergistic effect in promoting apoptosis of lung cancer cells.

The data show that the combination of dihydroartemisinin and chrysin with TOP2A inhibitor can increase the sensitivity of TOP2A inhibitor, reduce dosage and prolong treatment time. The pharmaceutical composition of the invention inhibits TOP2A expression based on dihydroartemisinin combined with chrysin, causes DNA damage, promotes lung cancer cell apoptosis, thereby improving the sensitivity of lung cancer cells to TOP2A inhibitors and enhancing the therapeutic effect of the drug.

The above-described embodiments are only preferred embodiments of the invention, and not all embodiments of the invention are possible. Any obvious modifications thereof, which would be apparent to those skilled in the art without departing from the principles and spirit of the present invention, should be considered to be included within the scope of the appended claims.

Claims

1. Application of dihydroartemisinin combined with chrysin in the preparation of products that increase the sensitivity of tumor cells to TOP2A inhibitors.

2. The use of dihydroartemisinin combined with chrysin according to claim 1 in the preparation of a product for increasing the sensitivity of tumor cells to TOP2A inhibitors, characterized in that the TOP2A inhibitor is specifically used to hinder DNA repair.

3. Use of dihydroartemisinin combined with chrysin according to claim 1 in the preparation of a product for increasing the sensitivity of tumor cells to TOP2A inhibitors, characterized in that the TOP2A inhibitor is etoposide.

4. The use of dihydroartemisinin combined with chrysin in the preparation of a product for improving the sensitivity of tumor cells to TOP2A inhibitors according to claim 1, characterized in that the concentration ratio of dihydroartemisinin to chrysin is dihydroartemisinin: chrysin = 20 μM: 25 μM or 11 μM: 30 μM.

5. Use of dihydroartemisinin combined with chrysin according to claim 1 in the preparation of a product for increasing the sensitivity of tumor cells to TOP2A inhibitors, characterized in that the tumor is lung cancer.

6. Use of dihydroartemisinin combined with chrysin according to claim 5 in the preparation of a product for improving the sensitivity of tumor cells to TOP2A inhibitors, characterized in that the lung cancer is small cell lung cancer and non-small cell lung cancer.

7. A pharmaceutical composition for increasing the sensitivity of tumor cells to TOP2A inhibitors, characterized in that it comprises a TOP2A inhibitor, dihydroartemisinin and chrysin.

8. The pharmaceutical composition for increasing the sensitivity of tumor cells to TOP2A inhibitors according to claim 7, characterized in that the TOP2A inhibitor in the pharmaceutical composition is etoposide, and the ratio of dihydroartemisinin: chrysin: etoposide is 20 μM: 25 μM: 5.22 μM or 11 μM: 30 μM: 5.221 μM.

9. The pharmaceutical composition for increasing the sensitivity of tumor cells to TOP2A inhibitors according to claim 7, characterized in that the pharmaceutical composition further comprises an excipient acceptable in the pharmaceutical field.