Drug containing elemene and kinase inhibitor

Abstract

Provided is a combined drug for treating cancers. The combined drug contains β-elemene, the combined drug is a single compound preparation or a combination of two separate preparations, and the combined drug effectively realizes the synergistic effect among drugs.

Description

A medicine comprising elemene and a kinase inhibitor Technical Field

[0001] The invention relates to a synergistic pharmaceutical composition for treating cancer and relates to the field of medicine. Background Art

[0002] Cancer is a major medical challenge worldwide, posing a serious threat to human health and life. Nearly 10 million people die from cancer annually worldwide, with approximately one million dying from malignant tumors in my country each year. Currently, cancer treatment primarily relies on surgery, chemotherapy, and radiotherapy. However, many chemotherapy drugs currently in clinical use still suffer from significant systemic side effects, drug resistance, and limited selectivity. In recent years, numerous clinical and experimental studies have demonstrated the effectiveness of Traditional Chinese Medicine (TCM) and combined drug therapies in tumor prevention, treatment, and recovery, and have gradually become a focus of attention.

[0003] β-elemene, a naturally occurring sesquiterpene and the primary component of zedoary turmeric oil and citronella oil, has been shown to be a broad-spectrum treatment for various tumors, demonstrating promising applications and market demand. To meet the treatment needs of cancer patients as quickly as possible, the development of more clinically valuable combination drug formulations and delivery methods is urgently needed. Summary of the Invention

[0004] The present invention provides a combination drug for treating cancer, wherein the combination drug comprises β-elemene and a second drug, wherein the second drug is a kinase inhibitor, and preferably the kinase inhibitor is selected from one or more of a TKI inhibitor, a KRAS inhibitor, an IDH1 inhibitor, and an mTOR inhibitor.

[0005] Furthermore, the TKI inhibitor is anlotinib.

[0006] Furthermore, the molar ratio of anlotinib to β-elemene is 1:0.1-1:2000, preferably 1:1-1:2000, preferably 1:1-1:1500, preferably 1:1-1:1282, preferably 1:10-1:1282, preferably 1:20-1:321, preferably 1:40-1:321, preferably 1:130-1:321.

[0007] Furthermore, the molar ratio of anlotinib to β-elemene is 1:0.1, 1:1, 1:10, 1:20, 1:40, 1:130, 1:321, 1:1282, 1:1500, and 1:2000, and preferably the molar ratio of anlotinib to β-elemene is 1:40, 1:130, and 1:321.

[0008] Furthermore, the TKI inhibitor is icotinib.

[0009] Furthermore, the molar ratio of icotinib to β-elemene is 1:25-1:1000, preferably 1:100-1:1000, preferably 1:25-1:400, preferably 1:100-1:400, preferably 1:100-1:200.

[0010] Furthermore, the molar ratio of icotinib to β-elemene is 1:25, 1:50, 1:100, 1:200, 1:400, or 1:1000, and preferably the molar ratio of icotinib to β-elemene is 1:100 or 1:200.

[0011] Furthermore, the TKI inhibitor is ceritinib.

[0012] Furthermore, the molar ratio of ceritinib to β-elemene is 1:7-1:222, preferably 1:28-1:111.

[0013] Furthermore, the molar ratio of ceritinib to β-elemene is 1:7, 1:14, 1:28, 1:56, 1:111, 1:222, and 1:500, and preferably the molar ratio of ceritinib to β-elemene is 1:28, 1:56, and 1:111.

[0014] Furthermore, the TKI inhibitor is larotrectinib.

[0015] Furthermore, the molar ratio of larotrectinib to β-elemene is 1:62.5-1:2000, preferably 1:125-1:1000, preferably 1:125-1:500, preferably 1:250-1:500.

[0016] Furthermore, the molar ratio of larotrectinib to β-elemene is 1:62.5, 1:125, 1:250, 1:500, 1:1000, or 1:2000, and preferably the molar ratio of larotrectinib to β-elemene is 1:250 or 1:500.

[0017] Furthermore, the TKI inhibitor is capmatinib.

[0018] Furthermore, the molar ratio of capmatinib to β-elemene is 1:0.5-1:20, preferably 1:0.5-1:5, preferably 1:1.25-1:20, preferably 1:1.25-1:5, preferably 1:2.5-1:5.

[0019] Furthermore, the molar ratio of capmatinib to β-elemene is 1:0.5, 1:1.25, 1:2.5, 1:5, or 1:20, and preferably the molar ratio of capmatinib to β-elemene is 1:2.5 or 1:5.

[0020] Furthermore, the TKI inhibitor is dacomitinib.

[0021] Furthermore, the molar ratio of dacomitinib to β-elemene is 1:1-1:2000, preferably 1:5-1:2000, preferably 1:5-1:500, preferably 1:5-1:250, preferably 1:5-1:125, preferably 1:15.63-1:2000, preferably 1:15.63-1:500, preferably 1:62.5-1:125.

[0022] Furthermore, the molar ratio of dacomitinib to β-elemene is 1:5, 1:15.63, 1:62.5, 1:125, 1:250, 1:500, and 1:2000.

[0023] Furthermore, the TKI inhibitor is apatinib.

[0024] Furthermore, the molar ratio of apatinib to β-elemene is 1:0.63-1:40, preferably 1:1.25-1:20, preferably 1:1.25-1:10, preferably 1:1.25-1:5, preferably 1:1.25-1:2.5.

[0025] Furthermore, the molar ratio of apatinib to β-elemene is 1:0.63, 1:1.25, 1:2.5, 1:5, 1:10, 1:20, and 1:40, and preferably the molar ratio of apatinib to β-elemene is 1:1.25 and 1:2.5.

[0026] Furthermore, the second drug is regorafenib.

[0027] Furthermore, the molar ratio of regorafenib to β-elemene is 1:12.5-1:1000, preferably 1:25-1:500, preferably 1:62.5-1:1000, preferably 1:125-1:500, preferably 1:250-1:500, preferably 1:12.5-1:200, preferably 1:25-1:100.

[0028] Furthermore, the molar ratio of regorafenib to β-elemene is 1:12.5, 1:25, 1:62.5, 1:100, 1:125, 1:200, 1:250, 1:500, and 1:1000.

[0029] Furthermore, the TKI inhibitor is gilteritinib.

[0030] Furthermore, the molar ratio of gilteritinib to β-elemene is 1:3.47-1:444.44, preferably 1:27.78.

[0031] Furthermore, the TKI inhibitor is neratinib.

[0032] Furthermore, the molar ratio of neratinib to β-elemene is 1:156.25-1:20000, preferably 1:5000.

[0033] Furthermore, the KRAS inhibitor is sotolacib.

[0034] Furthermore, the molar ratio of sotolacib to β-elemene is 1:10-1:1000, preferably 1:10-1:500, preferably 1:10-1:160, preferably 1:80-1:1000, preferably 1:80-1:500, preferably 1:80-1:160.

[0035] Furthermore, the molar ratio of sotolacib to β-elemene is 1:10, 1:40, 1:60, 1:80, 1:160, 1:500, and 1:1000.

[0036] Furthermore, the IDH1 inhibitor is ivosidenib.

[0037] Furthermore, the molar ratio of ivosidenib to β-elemene is 1:31.25-1:4000, preferably 1:250.

[0038] Furthermore, the mTOR inhibitor is temsirolimus.

[0039] Furthermore, the molar ratio of temsirolimus to β-elemene is 1:1250-1:10000, preferably 1:2500-1:5000, preferably 1:2500, 1:5000.

[0040] Furthermore, the combined drug is a pharmaceutical composition or a single compound preparation or a combination of two separate preparations. Preferably, the active ingredients of the pharmaceutical composition consist of β-elemene and a second drug.

[0041] Furthermore, the pharmaceutical composition comprises pharmaceutically acceptable excipients, including but not limited to injection oil, carriers, fillers, disintegrants, binders, lyoprotectants, pH regulators, preservatives, antioxidants, wetting agents, etc. The pharmaceutical composition can be prepared into a preparation, which can be one or more of tablets, capsules, pills, mixtures, gels, ointments, granules, powders, pastes, granules, pills, oral liquids, pills, injections or injections, or any other dosage form.

[0042] Furthermore, the present invention provides a use of a drug in preparing a drug for treating cancer.

[0043] Furthermore, the cancer is lung cancer, breast cancer, renal cell carcinoma, ovarian cancer, small cell lung cancer, non-small cell lung cancer, malignant melanoma, B lymphocyte tumor (including chronic lymphocytic leukemia / small lymphocytic lymphoma, mantle cell lymphoma, Waldenstrom's macroglobulinemia, follicular lymphoma, non-germinal center subtype diffuse large B-cell lymphoma, etc.), leukemia, colorectal cancer, malignant pleural mesothelioma, pancreatic cancer, head and neck cancer, gastric cancer, bladder cancer, bone cancer, esophageal cancer, brain tumor, liver cancer, cervical cancer, cancerous chest, ascites, preferably the cancer is lung cancer, liver cancer, colorectal cancer, leukemia, breast cancer, renal cell carcinoma, preferably the cancer is lung cancer, breast cancer, liver cancer, renal cell carcinoma, preferably lung cancer, breast cancer, renal cell carcinoma, preferably lung cancer, liver cancer, the lung cancer is preferably lung squamous cell carcinoma or large cell carcinoma.

[0044] Specific implementation examples

[0045] The technical solution of the present invention is further described in detail below in conjunction with the accompanying drawings and specific implementation examples, but the implementation of the present invention is not limited thereto. Equivalent replacements, combinations, improvements or modifications of the technical solution of the present invention made by those skilled in the art based on the description of the present invention should be included in the scope of protection of the present invention.

[0046] Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art. Unless otherwise specified, the reagents and materials used in the following examples are all commercially available.

[0047] The following examples are calculated in the following manner:

[0048] 1) Calculation of in vitro tumor growth inhibition rate

[0049] Tumor cell growth inhibition rate IR (%) = (1-(OD compound-OD blank control) / (OD vehicle control-OD blank control))*100%

[0050] 2) Determination of the efficacy of dual-drug combination therapy:

[0051] The determination was made using the Kim Jong-kyun q value method (see non-patent literature: Dai Tijun, "Addition in combined medication", Chinese Journal of Pharmacology, December 1980, 1(2): 70-76). q = P (A+B) / (P A +P B -P A ×P B ). Where P A 、P B and P (A+B)These are the inhibition rates for the β-elemene group, the second drug group, and the combination of the two drugs. A q < 0.85 indicates an antagonistic effect between the two drugs; a q > 1.15 indicates a synergistic effect; and a 0.85 ≥ q ≥ 1.15 indicates an additive effect.

[0052] 3) Calculation of in vivo tumor growth inhibition rate:

[0053] Tumor growth inhibition rate (TGI) (%) = [(1-(average bioluminescence signal at the end of dosing in a treatment group - average bioluminescence signal at the start of dosing in the treatment group)) / (average bioluminescence signal at the end of treatment in the vehicle control group - average bioluminescence signal at the start of treatment in the vehicle control group)] * 100%. The data in this application are all statistically consistent.

[0054] Example 1

[0055] β-elemene and anlotinib are divided into:

[0056] Group 1: 2000uM β-elemene, Group 2: 500uM β-elemene, Group 3: 62.5uM β-elemene, Group 4: 31.25uM β-elemene; Group 5: 15.625uM β-elemene, Group 6: 1.56uM anlotinib;

[0057] Group seven: 2000uM β-elemene + 1.56uM anlotinib, group eight: 500uM β-elemene + 1.56uM anlotinib, group nine: 62.5uM β-elemene + 1.56uM anlotinib, group ten: 31.25uM β-elemene + 1.56uM anlotinib, group eleven: 15.625uM β-elemene + 1.56uM anlotinib.

[0058] The above groups all used 0.25% DMSO and 0.25% anhydrous ethanol as solvents.

[0059] Cell Culture: NCI-H2170 cells were seeded in sterile cell culture flasks with an appropriate amount of 1640 medium supplemented with 10% FBS and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. The cells grew as an adherent monolayer and were passaged every 2-3 days. During passage, the cells were digested with 0.25% trypsin supplemented with 0.02% EDTA for 2-4 minutes. The culture medium was pipetted and perfused to create a cell suspension, which was then plated at the desired concentration. NCI-H2170 cells in the logarithmic growth phase were prepared as a single-cell suspension according to the passage method. The cells were counted and adjusted to a cell concentration of 3.5 x 10^4 cells / mL. The cell suspension was plated in a 96-well cell culture plate, with 100 μL of cell suspension added to each well. A well without cell culture medium was used as a blank for zeroing. After the culture plate was placed in a 37°C, 5% CO2 incubator for 12 hours, the culture medium in the well was removed, and 100 μL of culture medium (containing 2.5 μL of different concentrations of β-elemene + 2.5 μL of anlotinib) was added to groups 7 to 11, 100 μL of culture medium (containing 2.5 μL of different concentrations of β-elemene + 2.5 μL of DMSO) was added to groups 1 to 5, and 100 μL of culture medium (containing 2.5 μL of anhydrous ethanol + 2.5 μL of anlotinib) was added to group 6. The vehicle control was to add 100 μL of culture medium (containing 2.5 μL of DMSO and 2.5 μL of anhydrous ethanol). Three replicates were set up for each group. After 72 hours of culture, the culture medium was removed, 10% CCK8 working solution was added, and the cells were incubated at 37°C for 1 hour. The OD value was then measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 1:

[0060] Table 1

[0061] Example 2

[0062] β-elemene and icotinib are divided into:

[0063] Group 1: 2000uM β-elemene, Group 2: 1000uM β-elemene, Group 3: 500uM β-elemene, Group 4: 250uM β-elemene, Group 5: 125uM β-elemene; Group 6: 5uM icotinib;

[0064] Group VII: 2000uM β-elemene + 5uM icotinib, Group VIII: 1000uM β-elemene + 5uM icotinib, Group IX: 500uM β-elemene + 5uM icotinib, Group X: 250uM β-elemene + 5uM icotinib, Group XI: 125uM β-elemene + 5uM icotinib.

[0065] The above groups all used 0.25% DMSO and 0.25% anhydrous ethanol as solvents.

[0066] Cell Culture: NCI-H2170 cells were seeded in sterile cell culture flasks with an appropriate amount of 1640 medium supplemented with 10% FBS and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. The cells grew as an adherent monolayer and were passaged every 2-3 days. During passage, the cells were digested with 0.25% trypsin supplemented with 0.02% EDTA for 2-4 minutes. The culture medium was pipetted and perfused to create a cell suspension, which was then plated at the desired concentration. NCI-H2170 cells in the logarithmic growth phase were prepared as a single-cell suspension according to the passage method. The cells were counted and adjusted to a cell concentration of 3.5 x 10^4 cells / mL. The cell suspension was plated in a 96-well cell culture plate, with 100 μL of cell suspension added to each well. A well without cell culture medium was used as a blank for zeroing. After 12 hours of adherent culture in a 37°C, 5% CO2 incubator, the culture medium in the wells was removed, and 100 μL of culture medium (containing 2.5 μL of different concentrations of β-elemene + 2.5 μL of icotinib) was added to groups 7 to 11, 100 μL of culture medium (containing 2.5 μL of different concentrations of β-elemene + 2.5 μL of DMSO) was added to groups 1 to 5, and 100 μL of culture medium (containing 2.5 μL of anhydrous ethanol + 2.5 μL of icotinib) was added to group 6. The vehicle control was the addition of 100 μL of culture medium (containing 2.5 μL of DMSO and 2.5 μL of anhydrous ethanol). Three replicates were set up for each group, and after 72 hours of culture, the culture medium was removed, 10% CCK8 working solution was added, and the cells were incubated at 37°C for 1 hour. The OD value was then measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 2:

[0067] Table 2

[0068] Example 3

[0069] Beta-elemene and ceritinib are divided into:

[0070] Group 1: 2000uM β-elemene, Group 2: 1000uM β-elemene, Group 3: 500uM β-elemene, Group 4: 250uM β-elemene, Group 5: 125uM β-elemene; Group 6: 62.5uM β-elemene, Group 7: 9uM ceritinib;

[0071] Group eight: 2000uM β-elemene + 9uM ceritinib, group nine: 1000uM β-elemene + 9uM ceritinib, group ten: 500uM β-elemene + 9uM ceritinib, group eleven: 250uM β-elemene + 9uM ceritinib, group twelve: 125uM β-elemene + 9uM ceritinib, group thirteen: 62.5uM β-elemene + 9uM ceritinib.

[0072] The above groups all used 0.25% DMSO and 0.25% anhydrous ethanol as solvents.

[0073] Cell Culture: NCI-H2170 cells were seeded in sterile cell culture flasks with an appropriate amount of 1640 medium supplemented with 10% FBS and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. The cells grew as an adherent monolayer and were passaged every 2-3 days. During passage, the cells were digested with 0.25% trypsin supplemented with 0.02% EDTA for 2-4 minutes. The culture medium was pipetted and perfused to create a cell suspension, which was then plated at the desired concentration. NCI-H2170 cells in the logarithmic growth phase were prepared as a single-cell suspension according to the passage method. The cells were counted and adjusted to a cell concentration of 3.5 x 10^4 cells / mL. The cell suspension was plated in a 96-well cell culture plate, with 100 μL of cell suspension added to each well. A well without cell culture medium was used as a blank for zeroing. After the culture plate was placed in a 37°C, 5% CO2 incubator for 12 hours, the culture medium in the well was removed, and 100 μL of culture medium (containing 2.5 μL of different concentrations of β-elemene + 2.5 μL of ceritinib) was added to groups 8 to 13, 100 μL of culture medium (containing 2.5 μL of different concentrations of β-elemene + 2.5 μL of DMSO) was added to groups 1 to 6, and 100 μL of culture medium (containing 2.5 μL of anhydrous ethanol + 2.5 μL of ceritinib) was added to group 7. The vehicle control was the addition of 100 μL of culture medium (containing 2.5 μL of DMSO and 2.5 μL of anhydrous ethanol). Three replicates were set up for each group, and after 72 hours of culture, the culture medium was removed, 10% CCK8 working solution was added, and the cells were incubated at 37°C for 1 hour. The OD value was then measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 3:

[0074] Table 3

[0075] Example 4

[0076] Beta-elemene and sotolacib are divided into:

[0077] Group 1: 2000uM β-elemene, Group 2: 1000uM β-elemene, Group 3: 500uM β-elemene, Group 4: 125uM β-elemene; Group 5: 12.5uM sotolacib;

[0078] Group 6: 2000uM β-elemene + 12.5uM sotolacib, Group 7: 1000uM β-elemene + 12.5uM sotolacib, Group 8: 500uM β-elemene + 12.5uM sotolacib, Group 9: 125uM β-elemene + 12.5uM sotolacib.

[0079] The above groups all used 0.25% DMSO and 0.25% anhydrous ethanol as solvents.

[0080] Cell Culture: NCI-H2170 cells were seeded in sterile cell culture flasks with an appropriate amount of 1640 medium supplemented with 10% FBS and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. The cells grew as an adherent monolayer and were passaged every 2-3 days. During passage, the cells were digested with 0.25% trypsin supplemented with 0.02% EDTA for 2-4 minutes. The culture medium was pipetted and perfused to create a cell suspension, which was then plated at the desired concentration. NCI-H2170 cells in the logarithmic growth phase were prepared as a single-cell suspension according to the passage method. The cells were counted and adjusted to a cell concentration of 3.5 x 10^4 cells / mL. The cell suspension was plated in a 96-well cell culture plate, with 100 μL of cell suspension added to each well. A well without cell culture medium was used as a blank for zeroing. After the culture plate was placed in a 37°C, 5% CO2 incubator for 12 hours, the culture medium in the well was removed, and 100 μL of culture medium (containing 2.5 μL of different concentrations of β-elemene + 2.5 μL of sotracibin) was added to groups 6 to 9, 100 μL of culture medium (containing 2.5 μL of different concentrations of β-elemene + 2.5 μL of DMSO) was added to groups 1 to 4, and 100 μL of culture medium (containing 2.5 μL of anhydrous ethanol + 2.5 μL of sotracibin) was added to group 5. The vehicle control was to add 100 μL of culture medium (containing 2.5 μL of DMSO and 2.5 μL of anhydrous ethanol). Three replicates were set up for each group. After 72 hours of culture, the culture medium was removed, 10% CCK8 working solution was added, and the cells were incubated at 37°C for 1 hour. The OD value was then measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 4:

[0081] Table 4

[0082] Example 5

[0083] Beta-elemene and larotrectinib are divided into:

[0084] Group 1: 2000 μM β-elemene, Group 2: 1000 μM β-elemene, Group 3: 500 μM β-elemene, Group 4: 250 μM β-elemene, Group 5: 125 μM β-elemene; Group 6: 62.5 μM β-elemene, Group 7: 1 μM larotrectinib;

[0085] Group eight: 2000 μM β-elemene + 1 μM larotrectinib, group nine: 1000 μM β-elemene + 1 μM larotrectinib, group ten: 500 μM β-elemene + 1 μM larotrectinib, group eleven: 250 μM β-elemene + 1 μM larotrectinib, group twelve: 125 μM β-elemene + 1 μM larotrectinib, group thirteen: 62.5 μM β-elemene + 1 μM larotrectinib.

[0086] The above groups all used 0.25% DMSO and 0.25% anhydrous ethanol as solvents.

[0087] Cell Culture: Cells were seeded in sterile cell culture flasks with an appropriate amount of 1640 medium plus 10% FBS and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. Cells grew as adherent monolayers and were passaged every 2-3 days. During passaging, cells were first digested with 2 mL of 0.25% trypsin for 2-4 minutes, then neutralized with 2 mL of 1640 medium plus 10% FBS. The culture medium was pipetted to separate the cells into single cells, and then passaged at a density of 2-3 x 10^6 cells / flask. NCI-H2170 cells in the logarithmic growth phase were prepared as a single-cell suspension according to the passaging protocol, counted, and adjusted to a cell density of 3.5 x 10^4 cells / mL. The cell suspension was seeded into a 96-well cell culture plate, with 100 μL of the cell suspension added to each well. A blank well was used as a zero-well. After the culture plate was placed in a 37°C, 5% CO2 incubator for 24 hours, the culture medium in the wells was removed, and 100 μL of culture medium (containing 0.25 μL of different concentrations of β-elemene + 0.25 μL of larotrectinib) was added to groups 8 to 13, 100 μL of culture medium (containing 0.25 μL of different concentrations of β-elemene + 0.25 μL of DMSO) was added to groups 1 to 6, and 100 μL of culture medium (containing 0.25 μL of anhydrous ethanol + 0.25 μL of larotrectinib) was added to group 7. The vehicle control was the addition of 100 μL of culture medium (containing 0.25 μL of anhydrous ethanol and 0.25 μL of DMSO). Three replicates were set up for each group. After 72 hours of culture, the culture medium in the wells was removed, 10% CCK8 working solution was added, and the cells were incubated at 37°C for 1 to 3 hours. The OD value was measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 5:

[0088] Table 5

[0089] Example 6

[0090] β-elemene and capmatinib are divided into:

[0091] Group 1: 2000 μM β-elemene, Group 2: 500 μM β-elemene, Group 3: 250 μM β-elemene, Group 4: 125 μM β-elemene; Group 5: 100 μM capmatinib;

[0092] Group 6: 2000 μM β-elemene + 100 μM capmatinib, Group 7: 500 μM β-elemene + 100 μM capmatinib, Group 8: 250 μM β-elemene + 100 μM capmatinib, Group 9: 125 μM β-elemene + 100 μM capmatinib.

[0093] The above groups all used 0.25% DMSO and 0.25% anhydrous ethanol as solvents.

[0094] Cell Culture: NCI-H2170 cells were seeded in sterile cell culture flasks with an appropriate amount of 1640 medium plus 10% FBS and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. The cells grew as adherent monolayers and were passaged every 2-3 days. Cells were first digested with 2 mL of 0.25% trypsin for 2-4 minutes, then neutralized with 2 mL of 1640 medium plus 10% FBS. The cells were pipetted into single cells and passaged at a density of 2-3 x 10^6 cells / flask. NCI-H2170 cells in the logarithmic growth phase were prepared as single-cell suspensions according to the passage protocol, counted, and adjusted to a cell density of 3.5 x 10^4 cells / mL. The cell suspension was seeded into 96-well cell culture plates, with 100 μL of the suspension added to each well. Take blank culture medium as blank zero well. After the culture plate was placed in a 37 ° C, 5% CO2 incubator for 24 hours, the culture medium in the well was removed, and 100 μL culture medium (containing 0.25 μL of different concentrations of β-elemene + 0.25 μL capmatinib) was added to groups 6 to 9, 100 μL culture medium (containing 0.25 μL of different concentrations of β-elemene + 0.25 μL DMSO) was added to groups 1 to 4, and 100 μL culture medium (containing 0.25 μL of anhydrous ethanol + 0.25 μL capmatinib) was added to group 5. The solvent control was to add 100 μL culture medium (containing 0.25 μL of anhydrous ethanol and 0.25 μL of DMSO). Three replicates were set up for each group. After 72 hours of culture, the culture medium in the well was removed, 10% CCK8 working solution was added, and the cells were incubated at 37 ° C for 1 to 3 hours. The OD value was measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 6:

[0095] Table 6

[0096] Example 7

[0097] β-elemene and dacomitinib are divided into:

[0098] Group 1: 2000 μM β-elemene, Group 2: 500 μM β-elemene, Group 3: 250 μM β-elemene, Group 4: 125 μM β-elemene; Group 5: 62.5 μM β-elemene, Group 6: 15.625 μM β-elemene, Group 7: 1 μM dacomitinib;

[0099] Group eight: 2000 μM β-elemene + 1 μM dacomitinib, group nine: 500 μM β-elemene + 1 μM dacomitinib, group ten: 250 μM β-elemene + 1 μM dacomitinib, group eleven: 125 μM β-elemene + 1 μM dacomitinib, group twelve: 62.5 μM β-elemene + 1 μM dacomitinib, group thirteen: 15.625 μM β-elemene + 1 μM dacomitinib.

[0100] The above groups all used 0.25% DMSO and 0.25% anhydrous ethanol as solvents.

[0101] Cell Culture: NCI-H2170 cells were seeded in sterile cell culture flasks with an appropriate amount of 1640 medium plus 10% FBS and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. Cells grew as adherent monolayers and were passaged every 2-3 days. Cells were first digested with 2 mL of 0.25% trypsin for 2-4 minutes, then neutralized with 2 mL of 1640 medium plus 10% FBS. Cells were pipetted with culture medium to separate into single cells and passaged at a density of 2-3 x 10^6 cells / flask. NCI-H2170 cells in the logarithmic growth phase were prepared as single-cell suspensions according to the passage protocol, counted, and adjusted to a cell density of 3.5 x 10^4 cells / mL. This cell suspension was seeded into 96-well cell culture plates, with 100 μL of cell suspension added to each well. A well without cell culture medium was used as a blank. After the culture plate was placed in a 37°C, 5% CO2 incubator for 12 hours, the culture medium in the well was removed, and 100 μL of culture medium (containing 0.25 μL of different concentrations of β-elemene + 0.25 μL of dacomitinib) was added to groups 8 to 13, 100 μL of culture medium (containing 0.25 μL of different concentrations of β-elemene + 0.25 μL of DMSO) was added to groups 1 to 7, and 100 μL of culture medium (containing 0.25 μL of anhydrous ethanol + 0.25 μL of dacomitinib) was added to group 8. The vehicle control was to add 100 μL of culture medium (containing 0.25 μL of anhydrous ethanol and 0.25 μL of DMSO). Three replicates were set up for each group. After 72 hours of culture, the culture medium was removed, 10% CCK8 working solution was added, and the cells were incubated at 37°C for 1 hour. The OD value was then measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 7:

[0102] Table 7

[0103] Example 8

[0104] β-elemene and apatinib are divided into:

[0105] Group 1: 2000uM β-elemene, Group 2: 1000uM β-elemene, Group 3: 500uM β-elemene, Group 4: 250uM β-elemene, Group 5: 125uM β-elemene; Group 6: 62.5uM β-elemene, Group 7: 31.25uM β-elemene, Group 8: 50uM apatinib;

[0106] Group 9: 2000uM β-elemene + 50uM apatinib, Group 10: 1000uM β-elemene + 50uM apatinib, Group 11: 500uM β-elemene + 50uM apatinib, Group 12: 250uM β-elemene + 50uM apatinib, Group 13: 125uM β-elemene + 50uM apatinib, Group 14: 62.5uM β-elemene + 50uM apatinib, Group 15: 31.25uM β-elemene + 50uM apatinib;

[0107] The above groups all used 0.5% DMSO and 1% anhydrous ethanol as solvents.

[0108] Cell Culture: HuH-7 cells were seeded into sterile cell culture flasks with an appropriate amount of DMEM + 10% FBS medium and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. Cells grew as adherent monolayers and were passaged every 2-3 days. Cells were first digested with 2 mL of 0.25% trypsin for 2-4 minutes, then neutralized with 2 mL of DMEM + 10% FBS. Cells were pipetted with culture medium to separate into single cells and passaged at a density of 2-3 x 10^6 cells / flask. HuH-7 cells in the logarithmic growth phase were prepared as a single-cell suspension according to the passage method, counted, and adjusted to a cell density of 3.5 x 10^4 cells / mL. The cell suspension was seeded into 96-well cell culture plates, with 100 μL of cell suspension added to each well. Blank culture medium was used as a blank well. After the culture plate was placed in a 37°C, 5% CO2 incubator for 24 hours, the culture medium in the wells was removed, and 100 μL of culture medium (containing 1 μL of different concentrations of β-elemene + 0.5 μL apatinib) was added to groups 9 to 15, 100 μL of culture medium (containing 1 μL of different concentrations of β-elemene + 0.5 μL DMSO) was added to groups 1 to 7, and 100 μL of culture medium (containing 1 μL of anhydrous ethanol + 0.5 μL apatinib) was added to group 8. The vehicle control was to add 100 μL of culture medium (containing 1 μL of anhydrous ethanol and 0.5 μL DMSO). Three replicates were set up for each group. After 72 hours of culture, the culture medium in the wells was removed, 10% CCK8 working solution was added, and the cells were incubated at 37°C for 1 to 3 hours. The OD value was measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 8:

[0109] Table 8

[0110] Example 9

[0111] Beta-elemene and regorafenib are divided into:

[0112] Group 1: 2000uM β-elemene, Group 2: 1000uM β-elemene, Group 3: 250uM β-elemene, Group 4: 125uM β-elemene; Group 5: 10uM regorafenib;

[0113] Group 6: 2000uM β-elemene + 10uM regorafenib, Group 7: 1000uM β-elemene + 10uM regorafenib, Group 8: 250uM β-elemene + 10uM regorafenib, Group 9: 125uM β-elemene + 10uM regorafenib.

[0114] The above groups all used 0.5% DMSO and 1% anhydrous ethanol as solvents.

[0115] Cell Culture: HuH-7 cells were seeded in sterile cell culture flasks with an appropriate amount of DMEM + 10% FBS medium and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. Cells grew as adherent monolayers and were passaged every 2-3 days. Cells were first digested with 2 mL of 0.25% trypsin for 2-4 minutes, then neutralized with 2 mL of DMEM + 10% FBS. Cells were pipetted with culture medium to separate into single cells and passaged at a density of 2-3 x 10^6 cells / flask. HuH-7 cells in the logarithmic growth phase were prepared as a single-cell suspension according to the passage method, counted, and adjusted to a cell density of 4.5 x 10^4 cells / mL. The cell suspension was seeded into a 96-well cell culture plate, with 100 μL of cell suspension added to each well. A blank well was used as a zero-well. After the culture plate was placed in a 37°C, 5% CO2 incubator for 24 hours, the culture medium in the well was removed, and 100 μL of culture medium (containing 1 μL of different concentrations of β-elemene + 0.5 μL of regorafenib) was added to groups 6 to 9, 100 μL of culture medium (containing 1 μL of different concentrations of β-elemene + 0.5 μL of DMSO) was added to groups 1 to 4, and 100 μL of culture medium (containing 1 μL of anhydrous ethanol + 0.5 μL of regorafenib) was added to group 5. The vehicle control was to add 100 μL of culture medium (containing 0.5 μL of DMSO and 1 μL of anhydrous ethanol). Three replicates were set up for each group. After 72 hours of culture, the culture medium in the well was removed, 10% CCK8 working solution was added, and the cells were incubated at 37°C for 1 to 3 hours. The OD value was measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 9:

[0116] Table 9

[0117] Example 10

[0118] Beta-elemene and regorafenib are divided into:

[0119] Group 1: 2000uM β-elemene, Group 2: 1000uM β-elemene, Group 3: 500uM β-elemene, Group 4: 250uM β-elemene, Group 5: 125uM β-elemene; Group 6: 2uM regorafenib;

[0120] Group VII: 2000uM β-elemene + 2uM regorafenib, Group VIII: 1000uM β-elemene + 2uM regorafenib, Group IX: 500uM β-elemene + 2uM regorafenib, Group X: 250uM β-elemene + 2uM regorafenib, Group XI: 125uM β-elemene + 2uM regorafenib.

[0121] The above groups all used 0.5% DMSO and 1% anhydrous ethanol as solvents.

[0122] Cell Culture: HCT-15 cells were seeded in sterile cell culture flasks with an appropriate amount of 1640 medium supplemented with 10% FBS and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. Cells grew as adherent monolayers and were passaged every 2-3 days. During passaging, cells were first digested with 2 mL of 0.25% trypsin for 2-4 minutes, then neutralized with 2 mL of DMEM supplemented with 10% FBS. The culture medium was pipetted to separate the cells into single cells, and then passaged at a density of 3-4 x 10^6 cells / flask. HCT-15 cells in the logarithmic growth phase were prepared as a single-cell suspension according to the passage method, counted, and adjusted to a cell density of 3.5 x 10^4 cells / mL. This cell suspension was seeded into a 96-well cell culture plate, with 100 μL of the cell suspension added to each well. A well without cell culture medium was used as a blank. The culture plate was placed in a 37°C, 5% CO2 incubator for 12 hours of adherent culture. The culture medium in the wells was removed, and 100 μL of culture medium (containing 1 μL of different concentrations of β-elemene + 0.5 μL of regorafenib) was added to groups 7 to 11, 100 μL of culture medium (containing 1 μL of different concentrations of β-elemene + 0.5 μL of DMSO) was added to groups 1 to 5, and 100 μL of culture medium (containing 1 μL of anhydrous ethanol + 0.5 μL of regorafenib) was added to group 6. The vehicle control was the addition of 100 μL of culture medium (containing 0.5 μL of DMSO and 1 μL of anhydrous ethanol). Three replicates were set up for each group. After 72 hours of culture, the culture medium was removed, 10% CCK8 working solution was added, and the cells were incubated at 37°C for 1 hour. The OD value was then measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 10:

[0123] Table 10

[0124] Example 11

[0125] Beta-elemene and gilteritinib are divided into:

[0126] Group 1: 2000 μM β-elemene, Group 2: 1000 μM β-elemene, Group 3: 500 μM β-elemene, Group 4: 250 μM β-elemene, Group 5: 125 μM β-elemene; Group 6: 62.5 μM β-elemene, Group 7: 31.25 μM β-elemene, Group 8: 15.625 μM β-elemene, Group 9: 4.5 μM gilteritinib;

[0127] Group 10: 2000 μM β-elemene + 4.5 μM gilteritinib, Group 11: 1000 μM β-elemene + 4.5 μM gilteritinib, Group 12: 500 μM β-elemene + 4.5 μM gilteritinib, Group 13: 250 μM β-elemene + 4.5 μM gilteritinib, Group 14: 125 μM β-elemene + 4.5 μM gilteritinib, Group 15: 62.5 μM β-elemene + 4.5 μM gilteritinib, Group 16: 31.25 μM β-elemene + 4.5 μM gilteritinib; Group 17: 31.25 μM β-elemene + 4.5 μM gilteritinib;

[0128] The above groups all used 1% anhydrous ethanol and 0.25% DMSO as solvents.

[0129] Cell Culture: K562 cells were seeded in a cell culture dish and supplemented with an appropriate amount of 1640 medium supplemented with 10% FBS. Cells were grown in suspension and passaged every two days. During passage, cells were pipetted and mixed thoroughly. Two-thirds of the cell suspension was aspirated and the same volume of fresh medium was added to the cell culture dish. The cells were then incubated in a 37°C, 5% CO2, and saturated humidity incubator. K562 cells in the logarithmic growth phase were prepared as per the passage method and single-cell suspensions were prepared. Cell counts were counted and the cell concentration was adjusted to 3 × 105 cells / mL. The cell suspension was plated in a 96-well cell culture plate, and 100 μL of the cell suspension was added to each well. A blank well was then used as a zero-check. The plate was then placed in a 37°C, 5% CO2 incubator until ready for use. The culture supernatant was not removed from the plate. 100 μL of drug-containing medium from each group was added to each well. The wells were pipetted several times to ensure uniform drug mixing and to achieve the desired final test concentration. Then, groups 10 to 17 were added with 200 μL of culture medium per well (containing 2 μL of different concentrations of β-elemene + 0.5 μL of gilteritinib), groups 1 to 8 were added with 200 μL of culture medium per well (containing 2 μL of different concentrations of β-elemene + 0.5 μL of DMSO), group 9 was added with 200 μL of culture medium per well (containing 2 μL of anhydrous ethanol + 0.5 μL of gilteritinib), and the vehicle control was added with 200 μL of culture medium per well (containing 2 μL of anhydrous ethanol and 0.5 μL of DMSO). Three replicates were set up for each group. After 72 hours of culture, 20 μL of CCK8 detection reagent stock solution was added to each well, incubated at 37 ° C for 2.5 hours, and then the OD value was measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 11:

[0130] Table 11

[0131] Example 12

[0132] Beta-elemene and ivosidenib are divided into:

[0133] Group 1: 2000 μM β-elemene, Group 2: 1000 μM β-elemene, Group 3: 500 μM β-elemene, Group 4: 250 μM β-elemene, Group 5: 125 μM β-elemene; Group 6: 62.5 μM β-elemene, Group 7: 31.25 μM β-elemene; Group 8: 15.625 μM β-elemene, Group 9: 500 nM ivosidenib;

[0134] Group 10: 2000 μM β-elemene + 500 nM ivosidenib, Group 11: 1000 μM β-elemene + 500 nM ivosidenib, Group 12: 500 μM β-elemene + 500 nM ivosidenib, Group 13: 250 μM β-elemene + 500 nM ivosidenib, Group 14: 125 μM β-elemene + 500 nM ivosidenib, Group 15: 62.5 μM β-elemene + 500 nM ivosidenib, Group 16: 31.25 μM β-elemene + 500 nM ivosidenib, Group 17: 15.625 μM β-elemene + 500 nM ivosidenib.

[0135] The above groups all used 1% anhydrous ethanol and 0.25% DMSO as solvents.

[0136] Cell Culture: K562 cells were seeded in a cell culture dish and supplemented with an appropriate amount of 1640 medium supplemented with 10% FBS. Cells were grown in suspension and passaged every two days. During passage, cells were pipetted and mixed thoroughly. Two-thirds of the cell suspension was aspirated and the same volume of fresh medium was added to the cell culture dish. The cells were then incubated in a 37°C, 5% CO2, and saturated humidity incubator. K562 cells in the logarithmic growth phase were prepared as per the passage method and single-cell suspensions were prepared. Cell counts were counted and the cell concentration was adjusted to 3 × 105 cells / mL. The cell suspension was plated in a 96-well cell culture plate, and 100 μL of the cell suspension was added to each well. A blank well was then used as a zero-check. The plate was then placed in a 37°C, 5% CO2 incubator until ready for use. The culture supernatant was not removed from the plate. 100 μL of drug-containing medium from each group was added to each well. The wells were pipetted several times to ensure uniform drug mixing and to achieve the desired final test concentration. Then, groups ten to seventeen were respectively added with 200 μL culture medium per well (containing 2 μL of different concentrations of β-elemene + 0.5 μL ivoside), groups one to eight were respectively added with 200 μL culture medium per well (containing 2 μL of different concentrations of β-elemene + 0.5 μL DMSO), group nine was added with 200 μL culture medium per empty well (containing 2 μL anhydrous ethanol + 0.5 μL ivoside), and the vehicle control was added with 200 μL culture medium per well (containing 2 μL anhydrous ethanol and 0.5 μL DMSO). Three replicates were set up for each group. After 72 hours of culture, 20 μL of CCK8 detection reagent stock solution was added to each well, incubated at 37 ° C for 2.5 hours, and then the OD value was measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated according to the inhibition rate. The synergistic effect is shown in Table 12:

[0137] Table 12

[0138] Example 13

[0139] Beta-elemene and neratinib are divided into:

[0140] Group 1: 2000uM β-elemene, Group 2: 1000uM β-elemene, Group 3: 500uM β-elemene; Group 4: 62.5uM β-elemene, Group 5: 31.25uM β-elemene, Group 6: 15.625uM β-elemene, Group 7: 0.1uM neratinib;

[0141] Group eight: 2000uM β-elemene + 0.1uM neratinib, group nine: 1000uM β-elemene + 0.1uM neratinib, group ten: 500uM β-elemene + 0.1uM neratinib, group eleven: 62.5uM β-elemene + 0.1uM neratinib, group twelve: 31.25uM β-elemene + 0.1uM neratinib; group thirteen: 15.625uM β-elemene + 0.1uM neratinib.

[0142] The above groups all used 0.5% DMSO and 1% anhydrous ethanol as solvents.

[0143] Cell Culture: MCF-7 cells were seeded into sterile cell culture flasks with an appropriate amount of DMEM + 10% FBS medium and cultured in an incubator at 37°C, 5% CO2, and saturated humidity. Cells grew as adherent monolayers and were passaged every 2-3 days. Cells were first digested with 2 mL of 0.25% trypsin for 2-4 minutes, then neutralized with 2 mL of DMEM + 10% FBS. Cells were pipetted with culture medium to separate into single cells and passaged at a density of 2-3 x 10^6 cells / flask. MCF-7 cells in the logarithmic growth phase were prepared as single-cell suspensions according to the passage protocol, counted, and adjusted to a cell density of 3.5 x 10^4 cells / mL. The cell suspension was seeded into 96-well cell culture plates, with 100 μL of cell suspension added to each well. Blank culture medium was used as a blank well. The culture plate was placed in a 37°C, 5% CO2 incubator for 24 hours of adherent culture. The culture medium in the wells was removed, and 100 μL of culture medium (containing 1 μL of different concentrations of β-elemene + 0.5 μL of neratinib) was added to groups eight to thirteen, 100 μL of culture medium (containing 1 μL of different concentrations of β-elemene + 0.5 μL of DMSO) was added to groups one to six, and 100 μL of culture medium (containing 1 μL of anhydrous ethanol + 0.5 μL of neratinib) was added to group seven. The vehicle control was the addition of 100 μL of culture medium (containing 1 μL of anhydrous ethanol and 0.5 μL of DMSO). Three replicates were set up for each group. After 72 hours of culture, the culture medium in the wells was removed, 10% CCK8 working solution was added, and the cells were incubated at 37°C for 1 to 3 hours. The OD value was measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 13:

[0144] Table 13

[0145] Example 14

[0146] Beta-elemene and temsirolimus are divided into:

[0147] Group 1: 500uM β-elemene, Group 2: 250uM β-elemene, Group 3: 125uM β-elemene; Group 4: 62.5uM β-elemene, Group 5: 50nM temsirolimus;

[0148] Group 6: 500uM β-elemene + 50nM temsirolimus, Group 7: 250uM β-elemene + 50nM temsirolimus, Group 8: 125uM β-elemene + 50nM temsirolimus, Group 9: 62.5uM β-elemene + 50nM temsirolimus.

[0149] The above groups all used 0.5% DMSO and 1% anhydrous ethanol as solvents.

[0150] Cell Culture: 786-o cells were seeded into sterile cell culture flasks and supplemented with an appropriate amount of DMEM + 10% FBS medium. The cells were cultured in an incubator at 37°C, 5% CO2, and saturated humidity. The cells grew as adherent monolayers and were passaged every 2-3 days. During passaging, cells were first digested with 2 mL of 0.25% trypsin for 2-4 minutes, then neutralized with 2 mL of DMEM + 10% FBS. The culture medium was pipetted to separate the cells into single cells, and then passaged at a density of 2-3 x 10^6 cells / flask. HuH-7 cells in the logarithmic growth phase were prepared as a single-cell suspension according to the passaging protocol, counted, and adjusted to a cell density of 4.5 x 10^4 cells / mL. The cell suspension was seeded into 96-well cell culture plates, with 100 μL of the cell suspension added to each well. A blank well was used as a zero-well. The culture plate was placed in a 37°C, 5% CO2 incubator for 24 hours of adherent culture. The culture medium in the wells was removed, and 100 μL of culture medium (containing 1 μL of different concentrations of β-elemene + 0.5 μL of temsirolimus) was added to groups 6 to 9, 100 μL of culture medium (containing 1 μL of different concentrations of β-elemene + 0.5 μL of DMSO) was added to groups 1 to 4, and 100 μL of culture medium (containing 1 μL of anhydrous ethanol + 0.5 μL of temsirolimus) was added to group 5. The vehicle control was the addition of 100 μL of culture medium (containing 0.5 μL of DMSO and 1 μL of anhydrous ethanol). Three replicates were set up for each group. After 72 hours of culture, the culture medium in the wells was removed, 10% of CCK8 working solution was added, and the cells were incubated at 37°C for 1 to 3 hours. The OD value was measured at 450 nM using an automatic microplate reader. The inhibition rate was calculated according to the above formula. The results showed that each group had a certain inhibitory effect on the cells. The q value was calculated based on the inhibition rate. The synergistic effect is shown in Table 14:

[0151] Table 14

[0152] Example 15

[0153] Cell Culture: Human lung cancer NCI-H460-luc (NCI-H460, HTB-177) cells were cultured as monolayers in RPMI1640 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin solution at 37°C in a 5% CO2 incubator. Cells were routinely digested and passaged using trypsin-EDTA. When cell saturation reached 80%-90%, cell viability reached 97.90%, and the desired cell number was reached, cells were harvested, counted, and plated.

[0154] Cell inoculation and modeling: Female BALB / c nude mice were anesthetized with 1.25% avertin and immobilized after the animals were deeply anesthetized. The left upper limb of the mice was disinfected with 75% ethanol. The surface location corresponding to the lungs of the mice was located. A 1-2 cm incision was made using surgical scissors. 20 μL (2x106 NCI-H460-luc cells and 50% Matrigel) cell suspension was inoculated into the lung lobes of the mice. The injection site was checked to ensure no leakage. The skin was then surgically sutured and the mice were kept warm and observed until they fully recovered. To alleviate the pain of the animals, meloxicam was administered daily on the day of surgery and 24 and 48 hours after surgery.

[0155] Animal grouping: After inoculation, mice were intraperitoneally injected with 150 mg / kg luminescent pigment according to their body weight (10 μL / g). Ten minutes later, the mice were pre-anesthetized with a mixture of oxygen and isoflurane. After the mice were anesthetized, they were moved to the imaging chamber of IVIS (Lumina II) to begin bioluminescent signal detection. The bioluminescent signals in the animals and the generated image information were detected and recorded. Six days after the NCI-H460-luc model was inoculated, group dosing began when the average luminescence value reached 2.21E+07, with each group containing 4 mice. There were four groups in total:

[0156] Group 1) normal saline,

[0157] Group 2) Anlotinib tablets (marketed by Zhengda Tianqing Pharmaceutical)

[0158] Group 3) β-elemene albumin preparation (2.0 g β-elemene and 4.8 g soybean oil were taken to obtain the oil phase. 30 g of commercially available 20% human albumin was measured and diluted to 30 ml with an appropriate amount of water for injection to obtain the aqueous phase. The oil phase was added to the aqueous phase and sheared at 12,000 rpm using an online shearing machine to obtain colostrum. The colostrum was homogenized using a high-pressure microfluidizer at 18,000 psi to obtain homogenized milk. 25 g of commercially available 40% trehalose solution was added to the homogenized milk, and water for injection was added to a total of 100 g. The mixture was stirred evenly and filtered through a 0.22 μm microporous membrane to obtain the final milk. The final milk was freeze-dried to obtain the β-elemene albumin preparation for injection).

[0159] Group 4) anlotinib tablets + β-elemene albumin preparation (wherein the molar ratio of anlotinib to β-elemene was 1:130).

[0160] Use normal saline to prepare both preparations into solutions. Anlotinib tablets: Dissolve one tablet (12 mg / tablet) in normal saline, stir with a magnetic stirrer, dilute to 24.000 mL, and shake gently to obtain a 0.5 mg / mL solution. β-Elemene Albumin Injection: Dissolve β-Elemene Albumin Injection (200 mg / vial) in normal saline to a volume of 16.667 mL. Shake gently to obtain a 12 mg / mL solution.

[0161] The dosing regimen is shown in Table 15. After treatment, the tumor inhibition rate and q value were calculated based on the aforementioned in vivo inhibition rate. It was found that all groups achieved tumor inhibition, and the q value was 1.26, indicating a synergistic effect. The results are shown in Table 16.

[0162] Table 15 Dosage regimen

[0163] Table 16 Research results

Claims

1. A combined drug for treating cancer, characterized in that The combined drug comprises β-elemene and a second drug, and the second drug is a kinase inhibitor. Preferably, the kinase inhibitor is selected from one or more of TKI inhibitors, KRAS inhibitors, IDH1 inhibitors, and mTOR inhibitors.

2. The combined medicament according to claim 1, wherein The TKI inhibitor is anlotinib. Preferably, the molar ratio of anlotinib to β-elemene is 1:0.1 - 1:2000, preferably 1:1 - 1:2000, preferably 1:1 - 1:1500, preferably 1:1 - 1:1282, preferably 1:10 - 1:1282, preferably 1:20 - 1:321, preferably 1:40 - 1:321, preferably 1:130 - 1:

321. Preferably, the molar ratio of anlotinib to β-elemene is 1:40, 1:130, 1:

321.

3. The combination medicament according to claim 1, wherein The TKI inhibitor is icotinib. Preferably, the molar ratio of icotinib to β-elemene is 1:25 - 1:1000, preferably 1:100 - 1:1000, preferably 1:25 - 1:400, preferably 1:100 - 1:400, preferably 1:100 - 1:

200. Preferably, the molar ratio of icotinib to β-elemene is 1:100, 1:

200.

4. The combined drug according to claim 1, wherein The TKI inhibitor is ceritinib. Preferably, the molar ratio of ceritinib to β-elemene is 1:28 - 1:

111. Preferably, the molar ratio of ceritinib to β-elemene is 1:28, 1:56, 1:

111.

5. The combined medicine according to claim 1, wherein The TKI inhibitor is larotrectinib. Preferably, the molar ratio of larotrectinib to β-elemene is 1:62.5 - 1:2000, preferably 1:125 - 1:1000, preferably 1:125 - 1:500, preferably 1:250 - 1:

500. Preferably, the molar ratio of larotrectinib to β-elemene is 1:250, 1:

500.

6. The combined medicament according to claim 1, wherein The TKI inhibitor is capmatinib. Preferably, the molar ratio of capmatinib to β-elemene is 1:0.5 - 1:20, preferably 1:0.5 - 1:5, preferably 1:1.25 - 1:20, preferably 1:1.25 - 1:5, preferably 1:2.5 - 1:

5. Preferably, the molar ratio of capmatinib to β-elemene is 1:2.5, 1:

5.

7. The combined medicament according to claim 1, wherein The TKI inhibitor is dacomitinib. Preferably, the molar ratio of dacomitinib to β-elemene is 1:1 - 1:2000, preferably 1:5 - 1:2000, preferably 1:5 - 1:500, preferably 1:5 - 1:250, preferably 1:5 - 1:125, preferably 1:15.63 - 1:2000, preferably 1:15.63 - 1:500, preferably 1:62.5 - 1:

125. Preferably, the molar ratio of dacomitinib to β-elemene is 1:62.5, 1:

125.

8. The combined medicament according to claim 1, wherein The TKI inhibitor is apatinib. Preferably, the molar ratio of apatinib to β-elemene is 1:0.63 - 1:40, preferably 1:1.25 - 1:20, preferably 1:1.25 - 1:10, preferably 1:1.25 - 1:5, preferably 1:1.25 - 1:2.

5. More preferably, the molar ratio of apatinib to β-elemene is 1:1.25 and 1:2.

5.

9. The combination medicament according to claim 1, wherein The TKI inhibitor is regorafenib. Preferably, the molar ratio of regorafenib to β-elemene is 1:12.5 - 1:1000, preferably 1:25 - 1:500, preferably 1:62.5 - 1:1000, preferably 1:125 - 1:500, preferably 1:250 - 1:500, preferably 1:12.5 - 1:200, preferably 1:25 - 1:

100. More preferably, the molar ratio of regorafenib to β-elemene is 1:25, 1:62.5, 1:100, 1:125, 1:200, 1:250, 1:500, 1:1000.

10. The combined medicine according to claim 1, characterized in that The TKI inhibitor is gilteritinib. Preferably, the molar ratio of gilteritinib to β-elemene is 1:3.47 - 1:444.44, preferably 1:27.

78.

11. The combined medicament according to claim 1, characterized in that The TKI inhibitor is neratinib. Preferably, the molar ratio of neratinib to β-elemene is 1:156.25 - 1:20000, preferably 1:5000.

12. The combined medicament according to claim 1, wherein The KRAS inhibitor is sotorasib. Preferably, the molar ratio of sotorasib to β-elemene is 1:10 - 1:1000, preferably 1:10 - 1:500, preferably 1:10 - 1:160, preferably 1:80 - 1:1000, preferably 1:80 - 1:500, preferably 1:80 - 1:

160. More preferably, the molar ratio of sotorasib to β-elemene is 1:80 and 1:

160.

13. The combined medicine according to claim 1, characterized in that The IDH1 inhibitor is ivosidenib. Preferably, the molar ratio of ivosidenib to β-elemene is 1:31.25 - 1:4000, preferably 1:

250.

14. The combined medicament according to claim 1, wherein The mTOR inhibitor is temsirolimus. Preferably, the molar ratio of temsirolimus to β-elemene is 1:1250 - 1:10000, preferably 1:2500 - 1:5000. More preferably, the molar ratio of temsirolimus to β-elemene is 1:2500 and 1:5000.

15. The combination medicament according to any one of claims 1-14, characterized in that The combined drug is a pharmaceutical composition, or a single compound preparation, or a combination of two separate preparations. Preferably, the active ingredient of the pharmaceutical composition consists of β-elemene and a second drug. Preferably, the pharmaceutical composition contains pharmaceutically acceptable excipients.

16. Use of the drug according to any one of claims 1-15 in the preparation of a drug for treating cancer, preferably the cancer is lung cancer, breast cancer, renal cell carcinoma, ovarian cancer, small cell lung cancer, non-small cell lung cancer, malignant melanoma, B-lymphocyte tumor, leukemia, colorectal cancer, malignant pleural mesothelioma, pancreatic cancer, head and neck cancer, gastric cancer, bladder cancer, bone cancer, esophageal cancer, brain tumor, liver cancer, cervical cancer, cancerous pleural effusion and ascites, preferably the cancer is lung cancer, liver cancer, colorectal cancer, leukemia, breast cancer, renal cell carcinoma, preferably lung cancer, liver cancer, breast cancer, renal cell carcinoma, preferably lung cancer, breast cancer, renal cell carcinoma, and the lung cancer is preferably squamous cell lung cancer, large cell lung cancer.

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