A pharmaceutical composition for treating non-small cell lung cancer KRAS G12C mutation and application thereof

Abstract

The application provides a drug composition for treating non-small cell lung cancer KRAS G12C mutation and application thereof, and relates to the technical field of biological medicine. The drug composition comprises trametinib and artesunate, and the mass ratio of the two is 1:100. The drug composition of trametinib and artesunate can more significantly inhibit the growth of tumors than single drug, the drug resistance time is significantly prolonged, the side reaction of single drug is reduced, and the price is low. The drug composition can be used for preparing a novel therapeutic drug for treating non-small cell lung cancer KRAS G12C mutation, and has important clinical significance.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to a pharmaceutical composition for treating non-small cell lung cancer KRAS G12C mutation and application thereof. BACKGROUND

[0002] Among all pathological types of lung cancer, non-small cell lung cancer (NSCLC) is the most common, accounting for about 85%, and adenocarcinoma is the most common type of NSCLC, accounting for about 40% of lung cancer, and squamous cell carcinoma accounts for 20% to 30% of lung cancer.

[0003] Since the beginning of the 21st century, the targeted drug EGFR-TKI gefitinib applied in the clinic has significantly improved the survival rate of patients with advanced EGFR mutant sensitive lung adenocarcinoma, and the adverse reactions are mild. After 20 years of development, the focus of cancer treatment has gradually shifted to precision medicine, and targeted therapy has become one of the most important treatment methods for NSCLC, especially in most NSCLC patients who have lost the opportunity for surgery because they were found to have the disease in the middle and late stages.

[0004] At present, the oncogenic driver genes of NSCLC mainly include EGFR, ALK, KRAS, ROS1, MET, RET, BRAF, HER2, etc. In the Chinese Medical Association Clinical Diagnosis and Treatment Guidelines for Lung Cancer (2022 edition), the NSCLC must-check genes EGFR, ALK, ROS1, RET, BRAF V600E and MET14 exon skipping mutation have been recommended as class 1 evidence, and the extended genes include MET amplification or overexpression, HER2, KRAS and other gene mutations as class 2A recommended evidence. With the emergence of more and more new targeted drugs with high efficiency and low toxicity, the survival time of most patients has been prolonged, which further highlights the importance of gene detection. After identifying the mutation site, using reasonable targeted treatment strategies, maximizing the benefits of patients in precision treatment has always been the goal of human medicine.

[0005] RAS is the most frequently mutated oncogene, including KRAS, NRAS and HRAS three mutant subtypes, accounting for 86%, 11% and 3%, respectively. These genes encode four closely related proteins (KRAS4A, KRAS4B, NRAS, HRAS), of which KRAS is the most common in pancreatic cancer, colon and lung cancer. In NSCLC, KRAS gene mutations are more common in lung adenocarcinoma (20%-40%), and KRAS G12C is the most common subtype. Although the RAS family is the earliest discovered lung cancer driver gene, there is currently no widely used effective drug targeting its target in domestic clinical practice. However, with the development of KRAS G12C covalent inhibitors, KRAS gene has been transformed from "undruggable" to "targetable drug target", rekindling the scientific community's interest in KRAS target research.

[0006] In recent years, some small molecule inhibitors targeting KRAS G12C protein (sotorasib, MRTX849, JNJ-74699157 and LY3499446) have emerged in preclinical and phase I clinical studies. In the CodeBreak100 study, sotorasib (also known as AMG510, Sotorasib) developed by Amgen was used to treat 124 cases of KRAS G12C mutant NSCLC patients who had disease progression after immunotherapy or chemotherapy. The objective response rate was 37%, the disease control rate was 81%, the median remission duration was 11.1 months, the median progression-free survival time was 6.8 months, and the median overall survival was 12.5 months. This is the first KRAS G12C inhibitor to publish clinical trial results. So far, it is the only one approved worldwide, but sotorasib is expensive and has poor drug accessibility, and has not been widely used in clinical practice. The remaining drugs are currently in clinical trials. Therefore, for patients with KRAS G12C mutant lung cancer, it is still necessary to explore treatment options with better efficacy and / or fewer adverse reactions.

[0007] Artesunate is a derivative of artemisinin, which can kill malaria parasites and is used to treat malaria. As more and more people study artesunate, it is found that artesunate is not only limited to the treatment of malaria, but also can treat other diseases. Artesunate has the effects of anti-human cytomegalovirus, anti-hepatitis B virus, anti-hepatitis C virus, etc. It also has certain effect in the treatment of some cancers. Artesunate has strong inhibitory effect on human hepatoma cells, cervical cancer cells and nasopharyngeal carcinoma cells; artesunate can down-regulate the expression of CDK2, CDK4, CyclinDl and other genes, inhibit cell cycle, and thereby inhibit glioma cell proliferation; studies have shown that artesunate can be combined with sorafenib to improve the sensitivity of liver cancer treatment. However, there is little research on artesunate in lung cancer, and there is no report on KRAS G12C mutant NSCLC. In addition, the international single-target Mek inhibitor trametinib has been marketed, and its indication is malignant melanoma and combined with dasatinib mesylate capsules for metastatic NSCLC patients with BRAF V600 mutation. So far, there is no report on the application of the combination of artesunate and trametinib in the treatment of KRAS G12C mutant NSCLC. SUMMARY

[0008] In view of the technical problems existing in the prior art, the present application aims to provide a drug composition for treating non-small cell lung cancer KRAS G12C mutation and its application.

[0009] One of the purposes of the present application is to provide a drug composition for treating non-small cell lung cancer KRAS G12C mutation, which comprises trametinib and artesunate.

[0010] Preferably, the mass ratio of the trametinib and artesunate is 1:100.

[0011] The second purpose of the present application is to provide the use of a drug composition for treating non-small cell lung cancer KRAS G12C mutation in the preparation of a drug for treating non-small cell lung cancer KRAS G12C mutation.

[0012] Preferably, the drug is made of the drug composition and a pharmaceutically acceptable carrier.

[0013] Preferably, the pharmaceutically acceptable carrier is a sustained-release agent, a filler, a binder, a humectant, a disintegrant, an absorption promoter, an adsorption carrier, a surfactant or a lubricant.

[0014] Preferably, the preparation form of the drug is any one of injection, tablet, granule or capsule.

[0015] Preferably, the drugs and pharmaceutical compositions comprise at least one of the following effects:

[0016] (1) inhibiting the activity of KRAS G12C gene mutant lung cancer cells;

[0017] (2) inhibiting the proliferation of KRAS G12C gene mutant lung cancer cells;

[0018] (3) inhibiting the growth of KRAS G12C gene mutant lung cancer tumors.

[0019] Advantages of the present application:

[0020] The present application is a combination of trametinib and artesunate for treating non-small cell lung cancer KRAS G12C mutation.

[0021] Currently, through cell experiments and animal experiments, and from the two dimensions of proteomics and single cell analysis, it is confirmed that trametinib and artesunate can treat lung cancer KRAS G12C gene mutant tumor cells, but the combination of trametinib and artesunate joint drug intervention scheme can more significantly inhibit the growth of tumors than single drug intervention scheme, the drug resistance time is significantly prolonged, the side effects of single drug are reduced, and the price is cheap.

[0022] The drug combination intervention scheme described in the present application can be used to prepare a new therapeutic drug for treating KRAS G12C mutant non-small cell lung cancer, which is expected to replace sorafenib for treating lung cancer KRAS mutant patients, and has important clinical significance, providing a new treatment approach for clinical practice in KRAS G12C mutant lung cancer patients. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 -a relative cell activity-reaction curve of artesunate;

[0024] Figure 1 -b artesunate-cell viability analysis;

[0025] Figure 1 -c relative cell activity-reaction curve of trametinib;

[0026] Figure 1 -d trametinib-cell viability analysis;

[0027] Figure 2 -a synergistic inhibitory effect of artesunate and trametinib;

[0028] Figure 2 -b synergistic effect score of artesunate and trametinib;

[0029] Figure 3 Fluorescence image of cell proliferation after drug intervention;

[0030] Figure 4 Changes in tumor volume of mice after drug intervention;

[0031] Figure 5 Changes in body weight of mice after drug intervention;

[0032] Figure 6 Changes in survival rate of mice after drug intervention. DETAILED DESCRIPTION

[0033] The following further describes the present application in conjunction with specific examples and experimental examples. However, these examples are only intended to illustrate the present application and are not intended to limit the scope of the present application. The experimental methods in the following examples, if not otherwise specified, are generally performed under conventional conditions.

[0034] According to a first aspect of the present application, a pharmaceutical composition for treating non-small cell lung cancer KRAS G12C mutation is provided, wherein the pharmaceutical composition comprises trametinib and artesunate.

[0035] Trametinib is a mitogen-activated extracellular signal-regulated kinase that mainly acts on the Meki signaling pathway to inhibit cell growth and proliferation, thereby better inhibiting tumor development. Artesunate is an organic compound that mainly involves regulating the human immune system. The combination of trametinib and artesunate can activate the FCγ signaling pathway on macrophages in animals, thereby treating lung cancer patients with KRAS G12C gene mutation.

[0036] In a preferred embodiment of the present application, the trametinib and artesunate are in the form of solid powders, and the mass ratio of trametinib to artesunate is 1:100.

[0037] According to a second aspect of the present application, the use of a pharmaceutical composition for treating non-small cell lung cancer KRAS G12C mutation in the preparation of a medicament for treating non-small cell lung cancer KRAS G12C mutation is provided.

[0038] In a preferred embodiment of the present application, the medicament is made of the composition and a pharmaceutically acceptable carrier.

[0039] In a preferred embodiment of the present application, the pharmaceutically acceptable carrier is a sustained-release agent, a filler, a binder, a humectant, a disintegrant, an absorption promoter, an adsorption carrier, a surfactant, or a lubricant.

[0040] In a preferred embodiment of the present application, the medicament is in the form of an injection, a tablet, a granule, or a capsule.

[0041] In a preferred embodiment of the present application, the drugs and pharmaceutical compositions comprise at least one of the following effects:

[0042] (1) inhibiting the activity of KRAS G12C gene mutation lung cancer cells;

[0043] (2) inhibiting the proliferation of KRAS G12C gene mutation lung cancer cells;

[0044] (3) inhibiting the growth of KRAS G12C gene mutation lung cancer tumor.

[0045] Example 1

[0046] Inhibitory effect of the combination of trametinib and artemether on the activity of LLC cells of KRAS G12C gene mutation lung cancer cells.

[0047] Experimental materials

[0048] (1) Artemether (ART, purchased from Sigma Company, purity 99.99%, item number A3731-500MG)

[0049] (2) Trametinib (Meki, purchased from Selleck Company, purity 99.99%, item number S8830)

[0050] (3) LLC cells with KRAS G12C gene mutation, provided by Beijing National Center for Protein Science (Military Academy of Sciences).

[0051] (4) CCK-8 kit (Cell Counting Kit-8, provided by Beijing Jinpulai Biotechnology Co., Ltd.).

[0052] Experimental method

[0053] (1) Single-agent dose-response curve and cell viability analysis

[0054] The LLC cell line with KRAS G12C gene mutation was cultured in a cell incubator, and when the cells grew to about 80% of the culture dish, the LLC cells had optimal activity.

[0055] First, take three groups of 96-well plates, add 8000 optimally active LLC cells to each well, and culture until the cells adhere;

[0056] Set up a control group, i.e. no artemether or trametinib drug group (0 μM), only add 100 μL of culture medium.

[0057] Set up a blank group, i.e. do not add cells, only add 100 μL of the same volume of culture medium.

[0058] Two groups of experimental groups were set up. The first group of 96-well plates was added with gradient concentrations of artemether (0 μM, 10 μM, 20 μM, 50 μM, 100 μM, 150 μM, 200 μM, 250 μM, 300 μM and 400 μM), and the second group of 96-well plates was added with gradient concentrations of trametinib (0 μM, 1 μM, 10 μM, 20 μM, 40 μM, 80 μM, 100 μM, and 150 μM), and incubated for 24 hours.

[0059] The medium was aspirated, 100 μL of 10% CCK-8 incubation solution was added to each well, and after incubation at 37°C for 1 hour, the relative activity of LLC cells after intervention of different groups of drugs was detected according to the absorbance value (450 nm).

[0060] The experimental results are shown in Tables 1-2, and the IC50 curve is calculated by GraphPad Prism. Figure 1 a, c), the IC50 value of artemether is 83.4 μM, and the IC50 value of trametinib is 38.34 μM. Artemether 80 μM and trametinib 30 μM were selected as the cell intervention concentration, and LLC cells were intervened again according to the above operation, and the results are shown in Tables 3-4, Figure 1 (b, d) **** indicates p<0.0001, indicating that there is a significant difference between the two drug treatment groups and the control group, that is, trametinib and artemether have inhibitory effect on the activity of KRAS G12C gene mutant lung cancer LLC cells.

[0061] Table 1: Absorbance values of LLC cell lines inhibited by different concentrations of artemether

[0062] 0 (μΜ) 10 (μΜ) 20 (μΜ) 50 (μΜ) 100 (μΜ) 150 (μΜ) 200 (μΜ) 250 (μΜ) 300 (μΜ) 400 (μΜ) 0.954 0.819 0.823 0.765 0.42 0.318 0.26 0.224 0.233 0.228 0.912 0.822 0.84 0.713 0.529 0.321 0.279 0.232 0.236 0.229 1.039 0.854 0.879 0.727 0.511 0.319 0.245 0.23 0.227 0.229 0.945 0.862 0.869 0.666 0.5 0.373 0.243 0.224 0.224 0.229 0.807 0.88 0.772 0.704 0.504 0.329 0.258 0.223 0.226 0.226 0.868 0.844 0.75 0.592 0.474 0.307 0.233 0.211 0.213 0.228

[0063] Table 2: Absorbance values of LLC cell lines inhibited by different concentrations of trametinib

[0064] 0 (μΜ) 1 (μΜ) 10 (μΜ) 20 (μΜ) 40 (μΜ) 80 (μΜ) 100 (μΜ) 150 (μΜ) 1.073 1.101 1.18 1.007 0.6 0.175 0.176 0.149 1.073 0.888 1.168 0.945 0.668 0.194 0.175 0.171 1.115 0.924 1.075 0.979 0.54 0.183 0.17 0.168 1.151 0.984 1.009 0.864 0.528 0.187 0.173 0.155 1.029 1.102 1.094 0.974 0.563 0.177 0.163 0.157 1.214 1.087 1.052 0.888 0.525 0.162 0.156 0.153

[0065] Table 3: Inhibitory effect of artemether

[0066] Control 1.036018 0.990407 1.128326 1.026244 0.87638 0.942624 ART (80 μM) **** ]] 0.556109 0.67448 0.654932 0.642986 0.64733 0.614751

[0067] Table 4: Inhibitory effect of trametinib

[0068] Control 0.967393 0.967393 1.005259 1.037716 0.927724 1.094515 Meki (30 μM) **** ]] 0.540947 0.602254 0.486852 0.476033 0.507588 0.473328

[0069] (2) SynergyFinder method for evaluating the synergistic effect of double drugs

[0070] 96-well plates were prepared by adding 8000 LLC cells with optimal activity to each well, and incubating for 24 hours until the cells adhered. Based on the IC50 values of the two single drugs, appropriate concentration ranges and gradients of the double-drug combination were selected and added to the above 96-well plates. After 24 hours of incubation, the absorbance (OD value) of each well was measured, and the measured data in Table 5 was imported into SynergyFinder for analysis.

[0071] Inhibition rate = 1 - (OD value of drug group - OD value of blank group) / (OD value of control group - OD value of blank group) x 100%.

[0072] The experimental results are shown in Table 5. Figure 2 Figure 2 a shows that as the concentration of the double-drug combination increases, the activity of the tumor cells gradually decreases. Figure 2 b shows that in the combination range of 20 μM to 100 μM artesunate and 1 μM to 40 μM trametinib, the synergy score is higher, with an average of 14.87, indicating strong synergy, i.e., the combination is more effective than the single drug.

[0073] Table 5: Double-drug combination inhibits LLC cell line absorbance value

[0074]

[0075] Example 2

[0076] Proliferation inhibition effect of trametinib and artesunate combination on LLC cells of KRAS G12C gene mutant lung cancer cells.

[0077] Experimental materials

[0078] (1) Artesunate (ART, purchased from Sigma Company, purity 99.99%, item number A3731-500MG)

[0079] (2) Trametinib (Meki, purchased from Selleck Company, purity 99.99%, item number S8830)

[0080] (3) LLC cells with KRAS G12C gene mutation, provided by Beijing National Center for Protein Science (Military Academy of Sciences).

[0081] (4) BeyoClick EdU-594 cell proliferation detection kit (purchased from Biyun Tian Biotechnology Co., Ltd., product number: C0078S).

[0082] Experimental method

[0083] EDU fluorescence method for detecting cell proliferation:

[0084] ​The cells were divided into 4 groups, namely control group (Control), artesunate group (ART, 80 μM), trametinib group (Meki, 30 μM), artesunate + trametinib group (ART, 80 μM + Meki, 30 μM).

[0085] Artesunate powder was aliquoted into EP tubes and stored at -80°C. 1 mg of ART powder was mixed with 2.6013 ml of DMSO solution to prepare a 1 mM ART solution, which was aliquoted and stored. Similarly, 1 mg of Meki powder was mixed with 1.6250 ml of DMSO solution to prepare a 1 mM Meki solution, which was aliquoted and stored. According to the IC50 of the drug, 80 μl of 1 mM ART was added to 1 ml of DMEM medium to prepare 80 μM ART. Similarly, 30 μl of 1 mM ART was added to 1 ml of DMEM medium to prepare 30 μM Meki.

[0086] The cells were evenly plated in 6-well plates, and after 24 hours of adherent growth to about 80%, the drug was given to stimulate the preparation of 2X EdU working solution 20 μM. The 37°C preheated EdU working solution was added to the 6-well plate in equal volume, so that the final concentration of EdU in the 6-well plate was 1X, and the cells were incubated for another 2 hours.

[0087] Remove the DMEM medium and add 1 ml of 4% paraformaldehyde fixing solution P0099, and fix at room temperature for 15 minutes. Remove the fixing solution and wash the cells with 1 ml of washing solution per well for 3 times, 3-5 minutes each time. Remove the washing solution and add 1 ml of permeabilization solution containing 0.3% Triton X-100 per well, and incubate at room temperature for 10-15 minutes. Remove the permeabilization solution and wash the cells with 1 ml of washing solution per well for 1-2 times, 3-5 minutes each time.

[0088] Prepare the reaction solution and remove the washing solution in the previous step. Add 0.5 ml of reaction solution per well, gently shake the culture plate to ensure that the reaction mixture can evenly cover the sample, and incubate at room temperature for 30 minutes in the dark. Remove the reaction solution and wash with washing solution for 3 times, 3-5 minutes each time. Observe the number of cells under a fluorescence microscope and record, and the relative number of cells is calculated as the number of random fields of view under a light microscope (unit: "number"). Figure 3 and Table 6.

[0089] Table 6: Relative cell activity count (number) after drug intervention

[0090] Control Artesunate **** ]] Trametinib **** ]]> Artesunate + trametinib **** ]] 812 444 120 84 800 428 118 76 796 488 140 92 842 442 126 68 860 446 142 77 822 459 176 82

[0091] From Figure 3As shown in Table 6, compared with the control group, the number of tumor cells in both the artesunate and trametinib groups was significantly reduced. The number of cells in the artesunate + trametinib group was further reduced compared with the artesunate + trametinib alone group, indicating that the combination of the two drugs also has an inhibitory effect on the LLC cell line, and the inhibitory effect of the two drugs is more significant than that of each drug alone. **** indicates p < 0.0001, indicating that there is a significant difference between the two drug-treated groups and the control group.

[0092] In addition, the number of cells inhibited by a single drug is far less significant than that of the combination of two drugs, and this is not caused by the simple additive effect of the two drugs. This suggests that the combination of two drugs has a synergistic effect and significantly enhances the activity of inhibiting tumor cells compared to a single drug.

[0093] Example 3

[0094] Animal experiments testing the effect of trametinib and artesunate combination on inhibiting tumor growth

[0095] In the subcutaneous tumorigenesis (CDX) animal study, healthy adult C57BL / 6 male mice weighing 20-22 g were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. and housed in our PFS-grade laboratory for 1-2 weeks. LLC cells were then mixed with Matrigel and implanted subcutaneously in the right posterior axillary line of the back to explore the drug's mechanism of action. Sotolacib (AMG510), a drug targeting the KRAS G12C mutation in lung cancer, served as a positive control group.

[0096] Since the tumor grew to 100mm under the mouse skin 3 Mice were randomly divided into five groups: a control group (0.9% NaCl, gavage), a sotolacib group (AMG, 100 mg / kg, gavage), a trametinib group (Meki, 1 mg / kg, gavage), an artesunate group (ART, 100 mg / kg, gavage), and an artesunate + trametinib group (ART + Meki, with a combination of trametinib at 1 mg / kg and artesunate at 100 mg / kg). Each group consisted of 10 mice, where mg / kg refers to the dose per kg of mouse weight. The body weight of each mouse was monitored daily, and the drug dosage was determined based on body weight. For example, if each group of mice weighed approximately 20 g, approximately 2 mg of ART powder would be required for each mouse, and 20 mg of ART powder would be weighed for 10 mice. The powder was placed in a 5 ml EP tube and dissolved in 200 μl of DMSO. After thorough pipetting and mixing, the powder was slowly added to 1800 μl of DMEM medium. Each mouse was gavaged with approximately 200 μl of drug each time. Other drugs were prepared in the same manner.

[0097] The initial tumor volume and weight of mice were recorded before drug intervention. The tumor volume growth and weight changes of mice in different groups were measured every 2 days after drug intervention for 30 days. If the volume of mice in each group exceeded the ethical volume of 2000mm, the tumor volume and weight changes of mice in different groups were measured every 2 days. 3 The recording was terminated and the mice in this group were killed by CO2. The mice died of asphyxiation after being exposed to CO2 in the cage for about 2-3 minutes.

[0098] Tumor volume = long diameter × short diameter 2 / 2, calculate and count the volume growth of the tumor.

[0099] The experimental results are shown in Table 7, Table 8, Figure 4 and Figure 5 shown.

[0100] Table 7 Changes in tumor volume in mice after drug intervention (n=10, mm 3 )

[0101]

[0102]

[0103] Table 8 Changes in body weight of mice after drug intervention (n=10, mm 3 )

[0104]

[0105] The mouse survival (OS) model was constructed in the same way as above, with 7 mice in each group and a tumor volume of approximately 100 mm 3 Drug intervention was started. The survival period (end point) of the mice was determined when the tumor did not grow beyond the ethical volume (2000 mm 3 ) of mice died and the tumor was larger than 2000mm 3 However, the mice still survived, and the recording period was 18 days. The experiment was terminated when the number of mice in each group died (or the tumor volume exceeded 2000mm 3 ) is greater than 85%, and the experimental results are shown in Table 9 and Figure 6 shown.

[0106] Table 9 Effect of drug intervention on the survival of mice (d)

[0107]

[0108]

[0109] like Figure 4 As shown, when the mouse tumor grows to 100mm 3The mice in different groups were intervened with drugs to explore the effect of drugs on the tumor volume of mice. The tumor volume of each group grew slowly in the early stage, and the control group reached the ethical value on the 10th day. Compared with the control group, the tumor volume growth slowed down after the intervention of artemether, and there was a significant difference with the control group from the 8th day, but it grew faster than the sorafenib group, and the tumor volume reached the ethical value in the group on the 18th day. The tumor volume of the trametin group and the double-drug combination group gradually decreased in the early stage, and slowly grew from the 14th day. The tumor inhibition rate was significantly higher than that of the sorafenib group, and the tumor volume did not reach the ethical value during the observation period. In addition, on the 14th day, the growth curves of the trametin group and the double-drug combination group began to separate, and the tumor volume of the double-drug combination group was smaller than that of the double-drug combination group, and the growth curves gradually separated from the 24th day, with a statistically significant difference. It is suggested that even if trametin is resistant, the double-drug combination group still has a good effect on inhibiting tumor, and prolongs the drug resistance time.

[0110] Figure 5 As shown in the figure, the body weight of mice in different groups changed after daily drug intervention. Body weight change can indirectly reflect the health status of mice and adverse reactions of drugs. The body weight of mice changed stably after drug intervention in the early stage, indicating that the drug had almost no side effects, and the mice were healthy. The body weight of mice in the control group increased significantly from the 6th day due to tumor growth. The body weight of mice in the artemether group and the sorafenib group fluctuated within a certain range. The body weight of mice in the trametin group began to decrease significantly on the 8th day, although the tumor volume of mice was small at this time, but the drug induced immune response of the body and other significant effects on the body weight of mice, leading to the weight loss of mice. It is worth noting that the body weight of mice in the artemether + trametin group did not fluctuate significantly, and still maintained a stable range. It is indicated that although trametin has an effect on inhibiting tumor in mice, the toxicity caused by long-term drug use leads to significant weight loss of mice, and the body cannot tolerate it. Artemether has the effect of regulating immunity, and after combined application, the mice tolerate well.

[0111] Figure 6 As shown in the figure, the effect of different groups of drugs on the survival time of LLC model mice. The mice in each group grew normally in the first 8 days, and were active. On the 9th day, the mice in the control group began to "die", and on the 18th day, there was only one mouse left in the control group, and the "death" rate was more than 85%, and the experiment was terminated. At this time, there were 5 mice left in the sorafenib group, and 4 mice left in the artemether group. There were 7 mice left in the trametin group and the artemether + trametin group. The above results suggest that artemether and trametin have an inhibitory effect on tumor in mice, and the inhibitory effect is significantly improved after the combination of the two drugs. Compared with the sorafenib group, trametin and the double-drug combination group more significantly inhibit tumor growth, and the OS time of mice is the longest.

[0112] In combination with the above experiment, it can be known that although artemether and trametinib are applied alone, they have inhibitory effect on KRAS G12C lung adenocarcinoma cells, but the treatment effect is poor. If the two are applied in combination, not only the tumor inhibition effect is obtained, but also the inhibitory effect is more significant compared with the target drug AMG510. In addition, the combination of the two drugs can make up for the occurrence of cell toxicity and drug resistance caused by long-term use of trametinib.

[0113] The above experimental results provide a theoretical basis for the drug composition of artemether and trametinib for clinical treatment of non-small cell lung cancer KRAS G12C mutation, and have wide application prospect in the field of medicine.

[0114] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, a number of modifications or equivalent replacements can be made to the technical solutions, and these modifications or equivalent replacements should also be considered as the protection scope of the present application.

Claims

1. Use of a composition comprising trametinib and artesunate in the preparation of a medicament for treating non-small cell lung cancer KRAS G12C mutation.

2. Use according to claim 1, wherein The mass ratio of the trametinib and the artesunate is 1:

100.

3. The use according to claim 1, wherein The medicament is made of the composition of trametinib and artesunate and a pharmaceutically acceptable carrier.

4. Use according to claim 3, characterized in that, The pharmaceutically acceptable carrier is one or several of sustained-release agents, fillers, binders, humectants, disintegrants, absorption promoters, adsorption carriers, surfactants or lubricants.

5. Use according to claim 3, characterized in that, The preparation form of the medicament is any one of injections, tablets, granules or capsules.

6. The use according to claim 1, wherein The medicament comprises at least one of the following effects: (1) Inhibiting the activity of KRAS G12C gene mutation lung cancer cells; (2) Inhibiting the proliferation of KRAS G12C gene mutation lung cancer cells; (3) Inhibiting the growth of KRAS G12C gene mutation lung cancer tumor.

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