CN118787649A - A pharmaceutical composition for treating KRAS G12C mutation in non-small cell lung cancer and its application - Google Patents

Abstract

The present invention provides a pharmaceutical composition for treating KRAS G12C mutation in non-small cell lung cancer and its application, and relates to the field of biomedicine technology. The pharmaceutical composition comprises trametinib and artesunate, and the mass ratio of the two is 1:100. The trametinib and artesunate pharmaceutical composition inhibits tumor growth more significantly than a single drug, significantly prolongs drug resistance time, reduces single drug side effects, and is cheap. The pharmaceutical composition of the present invention can be used to prepare a new therapeutic drug for treating KRAS G12C mutation non-small cell lung cancer, which has important clinical significance.

Description

A drug composition for treating KRAS G12C mutation in non-small cell lung cancer and its application

Technical Field

The present invention belongs to the field of biomedicine technology, and specifically relates to a pharmaceutical composition for treating KRAS G12C mutation in non-small cell lung cancer and an application thereof.

Background Art

Among all pathological types of lung cancer, non-small cell lung cancer (NSCLC) is the most common, accounting for about 85%, while 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.

Since the beginning of the 21st century, the clinically used targeted drug EGFR-TKI gefitinib has significantly improved the survival rate of patients with advanced EGFR mutation-sensitive lung adenocarcinoma, with mild adverse reactions. After more than 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 treatments for NSCLC, especially for most NSCLC patients who are in the middle and late stages of the disease and have lost the opportunity for surgery.

At present, the main oncogenic driver genes of NSCLC include EGFR, ALK, KRAS, ROS1, MET, RET, BRAF, HER2, etc. In the Chinese Medical Association's Guidelines for Clinical Diagnosis and Treatment of Lung Cancer (2022 Edition), the must-test genes for NSCLC, EGFR, ALK, ROS1, RET, BRAF V600E, and MET14 exon skipping mutations have been included as Class 1 recommended evidence, and extended genes including MET amplification or overexpression, HER2, KRAS, etc., are included as Class 2A recommended evidence. With the emergence of more and more new, highly effective, and low-toxic targeted drugs, the survival time of most patients has been extended, which further highlights the importance of genetic testing. After identifying the mutation site, a reasonable targeted treatment strategy is adopted to maximize the benefits of patients in precision treatment, which has always been the treatment goal of human medicine.

RAS is the most commonly mutated oncogene, including three mutant subtypes of KRAS, NRAS and HRAS, accounting for 86%, 11% and 3% respectively. These genes encode four closely related proteins (KRAS4A, KRAS4B, NRAS, HRAS), of which KRAS is 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 its 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 and promotion of KRAS G12C covalent inhibitors, the KRAS gene has been transformed from "undruggable" to "targetable drug target", rekindling the scientific community's interest in the study of KRAS targets.

In recent years, some small molecule inhibitors that specifically target the KRAS G12C protein (sotorasib, MRTX849, JNJ-74699157, and LY3499446) have emerged in preclinical and phase I clinical studies. In the Phase II CodeBreak100 study, Sotorasib (also known as AMG510, Sotorasib) developed by Amgen was used to treat 124 patients with KRAS G12C mutant NSCLC whose disease progressed after immunotherapy or chemotherapy. The objective response rate was 37%, the disease control rate was 81%, the median duration of response 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 in the world, but Sotorasib is expensive and has poor drug accessibility, and is not widely used in clinical practice. The remaining drugs are currently in the clinical trial stage. Therefore, for patients with KRAS G12C mutant lung cancer, there is still a need to explore treatment options with better efficacy and/or fewer adverse reactions.

Artesunate is a derivative of artemisinin that can kill malarial parasites and is used to treat malaria. As more and more people study artesunate, they find that the effects of artesunate are not limited to the treatment of malaria, but can also treat other diseases. Artesunate has anti-human cytomegalovirus, anti-hepatitis B virus, anti-hepatitis C virus, etc. It also has a certain effect in the treatment of some cancers. Artesunate has a strong inhibitory effect on human liver cancer cells, cervical cancer cells and nasopharyngeal cancer cells; artesunate can downregulate the expression of genes such as CDK2, CDK4, CyclinD1, inhibit the cell cycle, and thus inhibit the proliferation of glioma cells; studies have shown that artesunate can be combined with sorafenib to improve the sensitivity of liver cancer treatment. However, artesunate has rarely been studied in lung cancer, and has not been reported in NSCLC with KRAS G12C mutation. In addition, trametinib, a Mek single-target inhibitor that has been marketed internationally, is indicated for malignant melanoma and is suitable for patients with metastatic NSCLC who are positive for BRAF V600 mutations when combined with dabrafenib mesylate capsules. So far, there have been no reports on the application of a drug combination of artesunate and trametinib in the treatment of NSCLC with KRAS G12C mutations.

Summary of the invention

In view of the technical problems existing in the prior art, the present invention aims to provide a pharmaceutical composition for treating KRASG12C mutation in non-small cell lung cancer and its application.

One of the objects of the present invention is to provide a pharmaceutical composition for treating KRAS G12C mutation in non-small cell lung cancer, wherein the pharmaceutical composition comprises trametinib and artesunate.

Preferably, the mass ratio of trametinib to artesunate is 1:100.

A second object of the present invention is to provide a pharmaceutical composition for treating KRAS G12C mutation in non-small cell lung cancer and its use in the preparation of a drug for treating KRAS G12C mutation in non-small cell lung cancer.

Preferably, the drug is made of a pharmaceutical composition and a pharmaceutically acceptable carrier.

Preferably, the pharmaceutically acceptable carrier is a sustained-release agent, a filler, a binder, a wetting agent, a disintegrant, an absorption enhancer, an adsorption carrier, a surfactant or a lubricant.

Preferably, the drug is in the form of an injection, tablet, granule or capsule.

Preferably, the drug and pharmaceutical composition have at least one of the following effects:

(1) Inhibit the activity of lung cancer cells with KRAS G12C gene mutation;

(2) Inhibit the proliferation of lung cancer cells with KRAS G12C gene mutation;

(3) Inhibit the growth of lung cancer tumors with KRAS G12C gene mutation.

Beneficial effects of the present invention:

The present invention provides a pharmaceutical composition for treating KRAS G12C mutation in non-small cell lung cancer, which is formed by combining trametinib and artesunate.

Currently, it has been confirmed through cell experiments and animal experiments as well as from the two dimensions of proteomics and single-cell analysis that both trametinib and artesunate can treat tumor cells with KRAS G12C gene mutations in lung cancer. However, the combined drug intervention regimen of trametinib and artesunate can more significantly inhibit tumor growth than the single-drug intervention regimen, significantly prolong the drug resistance time, reduce the side effects of single drugs, and is cheaper.

The drug combination intervention regimen described in the present invention can be used to prepare a new therapeutic drug for treating KRAS G12C mutation non-small cell lung cancer, and is expected to replace sotolacib to treat patients with KRAS mutation lung cancer. It has important clinical significance and provides a new treatment approach for clinical practice intervention in patients with KRAS G12C mutation lung cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1-a Relative cell activity-response curve of artemisinin;

Figure 1-b Artemisinin-cell viability analysis;

Figure 1-c Trametinib relative cell activity-response curve;

Fig. 1-d Trametinib-cell viability analysis;

Figure 2-a Synergistic inhibitory effect of artesunate and trametinib;

Figure 2-b Artesunate and trametinib dual drug synergy score;

Figure 3 Fluorescence images of cell proliferation after drug intervention;

Figure 4 Changes in mouse tumor volume after drug intervention;

Figure 5. Changes in mouse body weight after drug intervention;

Fig. 6 Changes in mouse survival rate after drug intervention.

DETAILED DESCRIPTION

The following is a combination of specific embodiments and experimental examples to further illustrate the present invention. However, these embodiments are limited to illustrating the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific experimental conditions are not specified are usually based on conventional conditions.

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

Trametinib is a mitogen-activated extracellular signal-regulated kinase that inhibits cell growth and proliferation mainly by acting on the Meki signaling pathway, thereby better inhibiting the development of tumors. Artesunate is an organic compound that mainly regulates human immunity. 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 mutations.

In a preferred embodiment of the present invention, both trametinib and artesunate are in the form of solid powder, and the mass ratio of trametinib to artesunate is 1:100.

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

In a preferred embodiment of the present invention, the medicine is prepared from a composition and a pharmaceutically acceptable carrier.

In a preferred embodiment of the present invention, the pharmaceutically acceptable carrier is a sustained-release agent, a filler, a binder, a wetting agent, a disintegrant, an absorption enhancer, an adsorption carrier, a surfactant or a lubricant.

In a preferred embodiment of the present invention, the drug is in the form of an injection, tablet, granule or capsule.

In a preferred embodiment of the present invention, the medicine and pharmaceutical composition have at least one of the following effects:

(1) Inhibit the activity of lung cancer cells with KRAS G12C gene mutation;

(2) Inhibit the proliferation of lung cancer cells with KRAS G12C gene mutation;

(3) Inhibit the growth of lung cancer tumors with KRAS G12C gene mutation.

Example 1

The inhibitory effect of the combination of trametinib and artesunate on the activity of LLC cells with KRAS G12C gene mutation in lung cancer cells.

Experimental Materials

(1) Artesunate (ART, purchased from Sigma, purity 99.99%, product number A3731-500MG)

(2) Trametinib (Meki, purchased from Selleck, purity 99.99%, product number S8830)

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

(4) CCK-8 kit (Cell Counting Kit-8, purchased from Beijing Jinpulai Biotechnology Co., Ltd.).

Experimental methods

(1) Single-drug dose-response curve and cell viability analysis

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

First, three sets of 96-well plates were taken, 8000 LLC cells with optimal activity were added to each well, and cultured until the cells adhered to the wall;

A control group was set up, i.e., a group without artesunate or trametinib (0 μM) and only 100 μL of culture medium was added.

A blank group was set up, that is, no cells were added, and only 100 μL of an equal volume of culture medium was added.

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

The culture medium was aspirated, and 100 μL of 10% CCK-8 incubation solution was added to each well. After incubation at 37° C. for 1 hour, the relative activity of LLC cells after intervention with different drug concentrations was detected based on the absorbance value (450 nm).

The experimental results are shown in Tables 1 and 2. GraphPad Prism calculated the IC50 curve (Figure 1a, c). The IC50 value of artesunate was 83.4μM, and the IC50 value of trametinib was 38.34μM. Artesunate 80μM and trametinib 30μM were selected as the cell intervention concentrations, and LLC cells were intervened again as described above. The results are shown in Tables 3 and 4. Figure 1 (b, d) **** indicates p<0.0001, indicating that there are significant differences between the two drug treatment groups and the control group, that is, trametinib and artesunate have an inhibitory effect on the activity of KRAS G12C gene mutant lung cancer cells LLC cells.

Table 1 Absorbance values of LLC cell lines inhibited by different concentrations of artesunate

0(μM) 10(μM) 20(μM) 50(μM) 100(μM) 150(μM) 200(μM) 250(μM) 300(μM) 400(μM) 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

Table 2 Absorbance values of LLC cell lines inhibited by different concentrations of trametinib

0(μM) 1(μM) 10(μM) 20(μM) 40(μM) 80(μM) 100(μM) 150(μM) 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

Table 3 Inhibitory effect of artesunate

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

Table 4 Inhibitory effect of trametinib

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

(2) SynergyFinder method to evaluate the combined synergistic effect of two drugs

8000 LLC cells with optimal activity were added to each well of a 96-well plate and cultured for 24 hours until the cells adhered to the wall. Based on the IC50 values of the two single drugs, a reasonable concentration range and gradient of a double-drug combination were selected and added to the above 96-well plate. After incubation for 24 hours, the absorbance (OD value) of each well was measured, and the measurement data Table 5 was imported into SynergyFinder for analysis.

Inhibition rate = 1-OD value of the drug-treated group-OD value of the blank group/OD value of the control group-OD value of the blank group) × 100%.

The experimental results are shown in Figure 2. Figure 2a shows that as the concentration of the two drugs gradually increases, the activity of tumor cells gradually decreases. Figure 2b shows that in the combination range of 20μM to 100μM artesunate and 1μM to 40μM trametinib, the synergistic effect score is high, with an average of 14.87, and has a strong synergistic effect, that is, the combined use is more effective than the single use.

Table 5 Absorbance values of LLC cell lines inhibited by dual drug combination

Example 2

The inhibitory effect of trametinib and artesunate combination on the proliferation of LLC cells with KRAS G12C gene mutation.

Experimental Materials

(1) Artesunate (ART, purchased from Sigma, purity 99.99%, product number A3731-500MG)

(2) Trametinib (Meki, purchased from Selleck, purity 99.99%, product number S8830)

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

(4) BeyoClick EdU-594 cell proliferation detection kit (purchased from Beyotime Biotechnology Co., Ltd., product number: C0078S).

Experimental methods

EDU fluorescence method to detect cell proliferation:

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

Artesunate powder was dispensed into EP tubes and frozen at -80°C. 1 mg ART powder and 2.6013 ml DMSO solution were mixed to prepare ART with a concentration of 1 mM, and then dispensed and frozen. Similarly, 1 mg Meki powder and 1.6250 ml DMSO solution were mixed to prepare Meki with a concentration of 1 mM, and then dispensed and frozen. According to the IC50 of the drug, ART 80 μM was prepared, that is, 80 μl 1mM ART was taken and added to 1 ml DMEM culture medium, and the final concentration was 80 μM. In the same way, Meki 30 μM was prepared by taking 30 μl 1mM ART and adding it to 1 ml DMEM culture medium, and the final concentration was 30 μM.

The cells were evenly plated in a 6-well plate. After 24 hours of attachment and growth to about 80%, drug stimulation was given. A 2X EdU working solution of 20 μM was prepared. The EdU working solution preheated at 37°C was added to the 6-well plate in equal volumes to make the final EdU concentration in the 6-well plate 1X. The cells were incubated for another 2 hours.

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

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

Table 6: Relative cell activity counts in each group after drug intervention (cells)

Control group 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

As can be seen from Figure 3 and Table 6, compared with the control group, the number of tumor cells in the artesunate group and the trametinib group was significantly reduced. The number of cells in the artesunate + trametinib group was further reduced compared with the single drug group, indicating that the combination of the two drugs also has an inhibitory effect on the LLC cell line, and the tumor cell inhibition effect after the combination of the two drugs is more significant than that of the single drug. **** indicates p < 0.0001, indicating that there are significant differences between the two drug treatment groups and the control group.

In addition, the number of cells inhibited by a single drug is far less significant than that by 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.

Example 3

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

In the tumor cell subcutaneous tumorigenesis (CDX) animal experiment, healthy adult C57BL/6 male mice were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., weighing 20-22g. After being raised in the PFS-level laboratory of our center for 1-2 weeks, LLC cells were mixed with matrix gel and implanted subcutaneously in C57BL/6 mice (on the right back axillary line) to explore the mechanism of action of the drug in mice. Sotolacib (AMG510) is a targeted drug for KRAS G12C mutation in lung cancer, which was used as the positive drug control group.

When the tumor grew to 100 ^mm3 under the skin of the mouse, it was randomly divided into 5 groups: control group 0.9% NaCl, gavage, sotolacib group (AMG, 100 mg/kg, gavage), trametinib group (Meki, 1 mg/kg, gavage), artesunate group (ART, 100 mg/kg, gavage), artesunate + trametinib group (ART + Meki, the trametinib dosage in the composition is 1 mg/kg, and the artesunate dosage is 100 mg/kg), 10 mice in each group, where mg/kg refers to the dosage of mice per kg weight. The weight of each group of mice was monitored daily, and the dosage of the drug was determined based on the weight. For example, if the weight of each group of mice is about 20g, 1 mouse needs about 2mg ART powder, and 10 mice need to weigh 20mg ART powder. The powder was placed in a 5ml EP tube, and 200μl of DMSO was added to dissolve it. After sufficient pipetting and mixing, it was slowly added to 1800μl of DMEM culture medium. Each mouse was gavaged with about 200 μl of drug each time. Other drugs were prepared in the same way.

The initial tumor volume and mouse weight were recorded before drug intervention. After drug intervention, the growth of tumor volume and weight of mice in different groups were measured every 2 days for 30 days. If the volume of mice in each group exceeded the ethical volume of 2000 ^mm3 , the recording was terminated and the mice in the group were killed by _CO2 . The mice died of suffocation after being connected to _CO2 in the cage for about 2-3 minutes.

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

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

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

Table 8 Changes in mouse body weight after drug intervention (n=10, mm ³ )

The mouse survival (OS) model was constructed in the same way as above, with 7 mice in each group. Drug intervention was started when the tumor volume was about 100 mm ^3. The criteria for mouse survival (experimental endpoint) refer to the death of mice whose tumor growth did not exceed the ethical volume (2000 mm ³ ) and the survival of mice with tumors larger than 2000 mm ^3. The recording period was 18 days. The termination of the experiment was determined when the number of mice in each group died (or the tumor volume exceeded 2000 mm ³ ) was greater than 85%. The experimental results are shown in Table 9 and Figure 6.

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

As shown in Figure 4, after the mouse tumor grew to ^100mm3, different groups of mice were intervened with drugs to explore the effect of drugs on the tumor volume of mice. In the early stage, the tumor volume of each group grew slowly, and the volume of the control group reached the ethical value on the 10th day. Compared with the control group, after the intervention of artesunate, the tumor volume growth slowed down, and there was a significant difference with the control group from the 8th day, but it grew faster than the sotolacib group, and the tumor volume in the group reached the ethical value on the 18th day. In the trametinib group and the double-drug combination group, the tumor volume gradually shrank in the early stage, and began to grow slowly on the 14th day. The tumor inhibition rate was significantly higher than that of the sotolacib 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 trametinib 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. The growth curves began to separate from the 24th day, with statistical differences. It suggests that even if trametinib is resistant, the double-drug combination group still has a good tumor inhibition effect and prolongs the drug resistance time.

As shown in Figure 5, the weight changes of mice in different groups after daily drug intervention. Weight changes can indirectly reflect the physical health of mice and adverse drug reactions. After the initial drug intervention, the weight of mice changed steadily, indicating that the drug had almost no side effects and the mice were healthy. Starting from the 6th day, the weight of mice in the control group increased significantly due to tumor growth. The weight of mice in the artesunate group and the sotolacib group fluctuated within a certain range. On the 8th day, the weight of mice in the trametinib group began to drop significantly. Although the tumor volume of mice was small at this time, the drug-induced immune response of the body significantly affected the weight of mice, causing the mice to become thin. It is worth noting that the weight of mice in the artesunate + trametinib group did not fluctuate significantly and remained in a stable range. It shows that although trametinib alone has the effect of inhibiting mouse tumors, the toxicity caused by long-term medication causes a significant decrease in the weight of mice, which the body cannot tolerate. Artesunate has the effect of regulating immunity, and after combined application, mice tolerate it well.

As shown in Figure 6, the effects of different groups of drug interventions on the survival of LLC model mice. In the first 8 days, the mice in each group grew normally 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. The "death" rate exceeded 85%, and the experiment was terminated. At this time, there were 5 mice remaining in the sotolacib group and 4 mice remaining in the artesunate group. There were 7 mice remaining in the metinib group and the artesunate + metinib group. The above results indicate that artesunate and metinib have inhibitory effects on mouse tumors. After the combination of the two drugs, the inhibitory effect is significant, and compared with the sotolacib group, the metinib and double-drug combination group more significantly inhibited tumor growth, and the mouse OS time was the longest.

Combined with the above experiments, it can be seen that although artesunate and trametinib alone have inhibitory effects on KRAS G12C lung adenocarcinoma cells, the therapeutic effect is poor. If the two are used in combination, they not only have a tumor-suppressing effect, but also have a more significant inhibitory effect compared with the targeted drug AMG510. In addition, the combined use of the two drugs can make up for the cytotoxicity and drug resistance caused by long-term use of trametinib alone.

The above experimental results provide a theoretical basis for the use of the pharmaceutical combination of artesunate and trametinib in the clinical treatment of KRAS G12C mutation in non-small cell lung cancer, and have broad application prospects in the field of medicine.

The above description is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several modifications or equivalent substitutions can be made to the technical solution without departing from the principle of the present invention. These modifications or equivalent substitutions should also be regarded as the protection scope of the present invention.

Claims (7)

1. A pharmaceutical composition for treating KRAS G12C mutation in non-small cell lung cancer, characterized in that the pharmaceutical composition comprises trametinib and artesunate. 2. The pharmaceutical composition according to claim 1, wherein the mass ratio of trametinib to artesunate is 1:100. 3. Use of the pharmaceutical composition according to claim 1 in the preparation of a drug for treating KRAS G12C mutation in non-small cell lung cancer. 4. The use of the pharmaceutical composition according to claim 3, characterized in that the medicine is made of the pharmaceutical composition and a pharmaceutically acceptable carrier. 5. The use of the pharmaceutical composition according to claim 4, characterized in that the pharmaceutically acceptable carrier is one or more of a sustained-release agent, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, an adsorption carrier, a surfactant or a lubricant. 6. The use of the pharmaceutical composition according to claim 3, characterized in that the drug is in the form of an injection, tablet, granule or capsule. 7. The use of the pharmaceutical composition according to claim 4, characterized in that the drug and the pharmaceutical composition have at least one of the following effects: (1) Inhibit the activity of lung cancer cells with KRAS G12C gene mutation; (2) Inhibit the proliferation of lung cancer cells with KRAS G12C gene mutation; (3) Inhibit the growth of lung cancer tumors with KRAS G12C gene mutation.