A drug combination for treating KRAS G12C mutant digestive system tumors

Abstract

The present invention belongs to the field of medicine, and discloses a drug combination for treating KRAS G12C mutant digestive system tumors, wherein the drug combination includes a KRAS G12C inhibitor and an AKT allosteric inhibitor. After multi-level exploration and demonstration, the present invention creatively proposes that the AKT allosteric inhibitor MK2206 can produce a higher synergistic effect with the KRAS G12C inhibitor, which is superior to the sotorasib and cetuximab drug combination currently being tested in clinical practice. From a mechanistic point of view, MK2206 can promote the expression of the ETV4 / DUSP6 axis, and then cooperate with sotorasib to exert the dephosphorylation effect of ERK1 / 2. It is worth noting that the function of AKT regulating the ETV4 / DUSP6 axis does not depend on its kinase activity and the classical PI3K / AKT signaling pathway, and AKT kinase inhibitors cannot achieve similar pharmacological effects and clinical efficacy. The present invention provides a new drug combination strategy and theoretical basis for the clinical practice of KRAS G12C inhibitors in digestive system tumors.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a drug combination for treating KRAS G12C mutant digestive system tumors. Background Art

[0002] The rat sarcoma proto-oncogene (RAS) family is the earliest discovered human oncogene family, including three major members: Kirsten rat sarcoma viral oncogene homolog (KRAS), Harvey rat sarcoma viral oncogene homolog (HRAS) and neuroblastoma RAS viral oncogene homolog (NRAS). The RAS proteins encoded by this family have the ability to hydrolyze guanosine triphosphate (GTP) and exchange nucleotides, and play a key role in normal cells by activating the mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) pathways. When the RAS protein mutates, its hydrolysis ability is inhibited and "locked" in the GTP-bound state, thereby continuously activating downstream pathways and leading to malignant cell proliferation and carcinogenesis. Clinically, about 20% of tumors have activating mutations of RAS proteins (mainly KRASG12D, KRASG12V, KRAS G12C and KRAS G13D), and in colorectal cancer, non-small cell lung cancer and pancreatic adenocarcinoma, the proportion of RAS mutations is as high as 50%, 40% and 90%, respectively. Therefore, directly targeting RAS proteins is the most ideal and effective treatment strategy for treating malignant tumors driven by RAS mutations.

[0003] Currently, there are two KRAS G12C inhibitors, sotorasib and adagrasib, approved by the U.S. Food and Drug Administration for the clinical treatment of specific tumors carrying KRAS G12C mutations. my country's National Medical Products Administration also approved the first domestically produced KRAS G12C inhibitor, fluzivac, for the clinical treatment of advanced non-small cell lung cancer in August 2024. However, a large number of clinical studies have shown that there are significant differences in the efficacy of these drugs in patients with non-small cell lung cancer, colorectal cancer, and pancreatic cancer. According to the results of the Codebreak100 and KRYSTAL-1 clinical trials, the objective response rates of sotorasib monotherapy in patients with non-small cell lung cancer, colorectal cancer, and pancreatic cancer were 37.1%, 9.7%, and 21%, respectively, and the objective response rates of adagrasib monotherapy in patients with non-small cell lung cancer and colorectal cancer were 42.9% and 19%, respectively. On the other hand, the objective response rate achieved by KRAS G12C targeted therapy was mainly composed of partial response rate (33.9% for non-small cell lung cancer, 9.7% for colorectal cancer, and 21% for pancreatic cancer), and very few patients showed complete remission (3.2% for non-small cell lung cancer, 0% for colorectal cancer, and 0% for pancreatic cancer); more than half of the patients showed stable disease (43.5% for non-small cell lung cancer, 73% for colorectal cancer, and 63% for pancreatic cancer) or continued progression, indicating that resistance of digestive system tumors to KRAS G12C inhibitors develops rapidly and widely.

[0004] Existing studies suggest that ERK1 / 2 reactivation is the main reason for digestive system tumors to develop resistance to KRAS G12C inhibitors, but its specific regulatory mechanism remains unclear. Publication Nos. CN118043049A, CN115279376A, CN114761012A, CN118574617A, CN116710094A, CN117120449A, CN115942936A, CN117615762A, CN112955137A and CN117529314A mainly introduce that KRAS upstream proteins such as ERRB family, FAK kinase, MEK kinase, SHP2 protein, SOS1 protein related inhibitors and immunomodulators (targeting programmed death receptor 1 and its ligands) can produce synergistic effects with KRAS G12C inhibitors, among which KRAS G12C inhibitors combined with epidermal growth factor receptor (EGFR) can produce synergistic effects with KRAS G12C inhibitors. The EGFR (EGFR) inhibitors cetuximab (clinical trial registration number: NCT05198934) or panitumumab (clinical trial registration number: NCT04185883) can increase the objective response rate of patients with colorectal cancer to 26.4%-30%, but there is still a gap compared with the efficacy of patients with non-small cell lung cancer. At the same time, the inventors' previous studies have shown that the above-mentioned combination regimen cannot produce a synergistic effect in the digestive system tumor model resistant to KRAS G12C inhibitors, nor can it explain the ERK1 / 2 reactivation phenomenon and its internal mechanism. Therefore, it is urgent to explore the specific regulatory mechanism of the rapid reactivation or even superactivation of ERK1 / 2 in digestive system tumors under the treatment of KRAS G12C inhibitors, and then develop new drug combination regimens to improve the clinical treatment efficacy and increase the treatment benefits of patients. Summary of the invention

[0005] The object of the present invention is to overcome at least one deficiency of the prior art and provide a drug combination for treating KRAS G12C mutant digestive system tumors.

[0006] The technical solution adopted by the present invention is: In a first aspect, the present invention provides a drug combination for treating KRAS G12C mutant digestive system tumors, wherein the drug combination comprises a KRAS G12C inhibitor and an AKT allosteric inhibitor.

[0007] In some examples, the AKT allosteric inhibitor is selected from MK2206, ARQ-092, TAS-117 or a pharmaceutically acceptable salt thereof, and its mechanism is to activate the expression of ETV4 / DUSP6 and then cooperate with the KRAS G12C inhibitor to dephosphorylate ERK1 / 2 protein.

[0008] In some examples, the AKT allosteric inhibitor is selected from MK2206 or a pharmaceutically acceptable salt thereof.

[0009] In some examples, the KRAS G12C inhibitor is selected from any one of sotorasib, adagrasib or fozeleta.

[0010] In some examples, the KRAS G12C mutation digestive system tumor diseases include colorectal cancer and pancreatic cancer.

[0011] In some examples, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient, wherein the excipient comprises at least one of a solvent, a filler, a lubricant, a disintegrant, a buffer, a solubilizing agent, an antioxidant, an antibacterial agent, an emulsifier, a binder or a suspending agent.

[0012] In some examples, the dosage form of the drug combination is intravenous injection, suppository, gel, foam, enteric solvent, decoction, mixture, syrup, granule, pill, capsule, gel or nanoformulation.

[0013] In a second aspect, the present invention provides use of a drug combination comprising a KRAS G12C inhibitor and an AKT allosteric inhibitor in the preparation of a drug for inhibiting KRAS G12C mutant digestive system tumors.

[0014] In some examples, the KRAS G12C inhibitor is selected from any one of sotorasib, adagrasib or fozeleta.

[0015] In some examples, the AKT allosteric inhibitor is selected from MK2206 or a pharmaceutically acceptable salt thereof.

[0016] In some examples, the KRAS G12C mutation digestive system tumor diseases include colorectal cancer and pancreatic cancer.

[0017] The beneficial effects of the present invention are: After multi-level exploration and demonstration, the present invention creatively proposes that the AKT allosteric inhibitor MK2206 can produce a high synergistic effect with the KRAS G12C inhibitor (the ZIP simulation scores in the three major drug resistance models are 29.356, 29.466 and 16.729, respectively), especially in the drug resistance model, its effect is better than the sotorasib and cetuximab drug combination currently being tested in the clinic. From a mechanistic point of view, MK2206 can promote the expression of the ETV4 / DUSP6 axis, and then cooperate with sotorasib to dephosphorylate ERK1 / 2. It is worth noting that the function of AKT regulating the ETV4 / DUSP6 axis does not depend on its kinase activity and the classical PI3K / AKT signaling pathway, and AKT kinase inhibitors cannot achieve similar pharmacological effects and clinical efficacy (the ZIP simulation scores in the three major drug resistance models are 12.347, 14.935 and 7.206, respectively). The present invention provides a new drug combination strategy and theoretical basis for the clinical practice of KRAS G12C inhibitors in digestive system tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The results are the test results of the killing ability of different drug combinations in Example 1 on parental and drug-resistant cells.

[0019] Figure 2 This is the effect of ETV4 overexpression on the drug sensitivity of sotooraisb-resistant cells in Example 2.

[0020] Figure 3 This is the effect of ETV4 overexpression on ERK1 / 2 phosphorylation in sotorasib-resistant cells in Example 2.

[0021] Figure 4 This is the effect of ETV4 overexpression on DUSP6 expression in sotorasib-resistant cells in Example 2.

[0022] Figure 5 This is the effect of sotorasib stimulation at different time periods on the AKT signaling pathway of each parental cell in Example 2.

[0023] Figure 6 This is the effect of various inhibitors of the PI3K / AKT / mTOR pathway on ETV4 expression in Example 2.

[0024] Figure 7 Scoring the in vitro validation experiment in Example 3 (ZIP model).

[0025] Figure 8 This is the in vivo validation experiment evaluation in Example 3. DETAILED DESCRIPTION

[0026] The following disclosure provides many different embodiments or examples for implementing different solutions of the present invention.

[0027] Example 1: KRAS G12C inhibitor combined with epidermal growth factor receptor inhibitor has no synergistic effect in drug-resistant models Colorectal cancer parental cells SW837 and LIM2099 and their paired drug-resistant cells SW837 / KIR (deposited in the China Type Culture Collection at Wuhan University Collection Center, Wuchang District, Wuhan City, Hubei Province, with a deposit number of CCTCC NO: C2024360, classified as: human colorectal cancer cell SW837 / KIR Homo sapiens, and a deposit date of October 24, 2024) and LIM2099 / KIR (deposited in the China Type Culture Collection at Wuhan University Collection Center, Wuchang District, Wuhan City, Hubei Province, with a deposit number of CCTCC NO: C2024361, classified as: human colorectal cancer cell LIM2099 / KIR Homo sapiens, and a deposit date of October 24, 2024) were cultured in DMEM and RPMI-1640 culture media containing 10% fetal bovine serum, respectively, and placed at 37°C, 5% Incubate in a CO2-free cell culture incubator and change the medium every other day. When the cell density reaches 80%-90%, digest with 0.25% trypsin (containing 0.02% EDTA) and subculture at 5×10 3 The cells were seeded into 96-well plates at a density of 100 cells / well and placed in a cell incubator for further culture. After the cells adhered to the plate (24 hours after seeding), the fresh culture medium was replaced and the cells were divided into a blank control group, a sotorasib (MCE, Catalog No. HY114277) monotherapy group (the treatment concentration of SW837 and SW837 / KIR was 100nM; the treatment concentration of LIM2099 and LIM2099 / KIR was 1uM), a cetuximab (Merk, Catalog No. SJ20171039) monotherapy group (the treatment concentration was 50ug / ml) and a sotorasib+cetuximab drug combination group (three replicates were set for each group) and treated with the corresponding drugs and placed in a cell incubator for further culture for 72 hours. After the treatment, the cell culture medium in the 96-well plate was aspirated and a mixture of CCK8 (Cell Counting Kit-8) detection reagent (Dongaren, catalog number CK04) and cell culture medium was prepared in a ratio of 1:9 (prepared and used immediately). Then, 100ul of the above-prepared mixture was added to each well and the plate was placed back in the cell incubator for incubation for 1-3 hours. After the incubation, the absorbance of each well was detected using an ELISA reader (450nm wavelength) and converted into cell viability. The conversion formula is: Cell survival rate = [(experimental well-blank well) / (control well-blank well)] × 100%.

[0028] Example Figure 1 It can be seen that although in SW837 and LIM2099 parental cells, the combination of sotorasib + cetuximab drugs can increase the killing rate by 13.6% ( p <0.05), however, in SW837 / KIR and LIM2099 / KIR-resistant cells, the drug combination did not significantly improve the killing effect: the inhibition rates of sotorasib and cetuximab alone on SW837 / KIR were 15.73% and 9.76%, respectively, and the inhibition rate of the combined regimen was 19.06% ( p >0.05); the inhibition rates of sotorasib and cetuximab alone on LIM2099 / KIR were 19.88% and 11.73%, respectively, and the inhibition rate of the combination regimen was 23.14% ( p >0.05). The above results suggest that it is necessary to further reveal the mechanism of drug resistance and explore more efficient combination schemes.

[0029] Example 2: Exploration of the resistance mechanism of KRAS G12C inhibitors in gastrointestinal tumors This example verifies the biological functions and drug resistance regulatory mechanisms of differentially expressed genes (among which the differentially expressed gene ETS variant transcription factor 4 (ETS variant transcription factor 4, ETV4 ) is obtained through paired transcriptome sequencing and analysis. The sequencing and analysis methods are not specific and will not be elaborated on. The specific evaluation methods are as follows: (1) Construction of ETV4 overexpression plasmid: Firstly, the information of ETV4 transcripts was queried through the website of the National Center for Bioinformation (NCBI). The common transcript (NCBI number: NM_001079675.5) and protein (NCBI number: NP_001073143.1) were selected as the objects. The coding sequence of ETV4 protein was obtained by reverse transcription and in vitro polymerase chain reaction (PCR). It was then cloned into the lentiviral vector plasmid pLV3-CMV-MCS-3×FLAG-CopGFP-Puro. Finally, after molecular biological operations such as transformation, shaking, and plasmid extraction, the ETV4 overexpression plasmid pLV3-CMV-ETV4-3×FLAG-CopGFP-Puro was obtained.

[0030] (2) Lentiviral packaging and construction of ETV4 protein stable overexpression cell line: 293T cells in the logarithmic growth phase were inoculated in a 10 cm cell culture dish at a density of 60%-80%. ETV4 overexpression plasmid pLV3-CMV-ETV4-3×FLAG-CopGFP-Puro 6.63ug, packaging plasmid PsPAX 5.05ug and pMD2.G 3.32ug were co-transfected into 293T cells using lipo3000 transfection reagent. The transfected 293T cells were cultured in a cell culture incubator for 48 hours, and the culture medium was collected and filtered through a 0.45um filter to filter the cells and debris (to prevent cross-contamination between cells). The filtrate obtained was the lentiviral solution. During this period, the SW837 / KIR and MIA PaCa2 / KIR cells to be infected in the logarithmic growth phase (deposited in the China Type Culture Collection of Wuhan University Collection Center, Wuchang District, Wuhan City, Hubei Province, with a deposit number of CCTCC NO: C2024358, classified as: human pancreatic cancer cell MIAPaCa2 / KIR Homo sapiens, and a deposit date of October 24, 2024) were inoculated into a 6 cm cell culture dish one day before infection. During lentiviral infection, the original culture medium was discarded, 2 ml of fresh culture medium and 2 ml of filtered virus solution were added, and 3.2 ul of polybrene was added to promote virus infection. After 24 hours, the virus solution was discarded and fresh culture medium was replaced for continued culture. After 48 hours of infection, fresh culture medium was replaced and an appropriate concentration (1-2 ug / ml) of puromycin was added for resistance screening. Cells that still survived after 2-4 days were ETV4 stable overexpression cell lines, and the ETV4 expression level was detected by WB.

[0031] (3) Verification of ETV4 biological function and drug resistance regulation mechanism: First, the CCK8 method was used to detect whether the sensitivity of the drug-resistant strain with stable overexpression of ETV4 to sotorasib was restored. The specific operation process was the same as Example 1. Briefly, blank and ETV4-overexpressing SW837 / KIR and MIA PaCa2 / KIR drug-resistant cells were cultured at 5×10 3 The cells were inoculated at a density of 100 nM / well into a 96-well plate, and a blank control group and a sotorasib gradient treatment group (the treatment concentration was 100 nM and 1000 nM) were set up. The corresponding drugs were added for treatment and the cells were placed in a cell incubator for further culture for 72 hours. After the treatment time, the microplate reader readings of each well were detected and the cell survival rate of each group was analyzed.

[0032] Test results such as Figure 2 As shown: Compared with blank SW837 / KIR cells, the inhibition rate of SW837 / KIR cells overexpressing ETV4 increased by 23.3% after treatment with the same concentration of sotorasib ( p<0.05); similarly, compared with blank MIAPaCa2 / KIR cells, the inhibition rate of MIA PaCa2 / KIR cells overexpressing ETV4 increased by 20.6% after treatment with the same concentration of sotorasib ( p <0.05).

[0033] Furthermore, we verified whether the reversal of drug resistance by ETV4 overexpression was related to ERK1 / 2 dephosphorylation. The specific steps were as follows: S1) Blank and ETV4-overexpressing SW837 / KIR and MIA PaCa2 / KIR-resistant cells were seeded into 6-well plates at a density of 30%-40%, respectively. A blank control group and sotorasib gradient treatment group (treatment concentrations of 100 nM and 1000 nM) were set up and treated with corresponding drugs. The cells were placed in a cell incubator and cultured for 72 hours. S2) After the treatment time is over, the cells of each group are collected into 1.5 ml EP tubes, washed twice with phosphate buffered saline (PBS), and then added with protein lysis buffer containing 1× protease inhibitor and phosphatase inhibitor. After lysis on ice for 30 minutes, centrifuge at 12,000 rpm and 4°C for 10 minutes, collect the supernatant and perform BCA (Bicinchoninic Acid) quantification; S3) Add 5× protein loading buffer according to the sample volume and mix well, incubate at 100°C for 10 minutes. Polyacrylamide gel electrophoresis (10% electrophoresis gel, electrophoresis conditions are constant voltage 80V, adjusted to 100V after 30 minutes and maintained for 90 minutes) is used for protein separation; S4) After the electrophoresis is completed, transfer the membrane (the transfer condition is a constant current of 220 mA for 120 minutes); S5) After transfer, place the membrane in 8% skim milk solution and block at room temperature for 1 hour; after blocking, wash the membrane three times with 1×TBST buffer (Tris-buffered saline containing Tween 20), cut the required bands, add ERK1 / 2 (CST, Catalog No. 4695) and p-ERK1 / 2 antibodies (to detect ERK1 / 2 phosphorylation level and thus reflect activation level, CST, Catalog No. 4370), respectively, and place it in a shaker at 4°C overnight; S6) On the second day, the primary antibody was recovered and the membrane was washed three times with 1×TBST buffer for 10 minutes each time. The secondary antibody was incubated at room temperature for 1 hour, and then the membrane was washed three times with TBST buffer for 10 minutes each time. After washing, the membrane was developed chemically.

[0034] Test results such as Figure 3As shown: Compared with blank SW837 / KIR and LIM2099 / KIR cells, the phosphorylation levels of ERK1 / 2 in SW837 / KIR and LIM2099 / KIR cells overexpressing ETV4 were significantly downregulated after treatment with the same concentration of sotorasib. The above results confirm the powerful function of ETV4 in inhibiting ERK1 / 2 phosphorylation reactivation and reversing KRAS G12C inhibitor resistance.

[0035] Next, the upstream and downstream regulatory pathways and effector proteins of ETV4 were further explored. The inventors found that the expression level of dual specificity phosphatase 6 (DUSP6), which can directly negatively regulate the phosphorylation level of ERK1 / 2, was downregulated in the drug-resistant model. Then, the total protein in blank and ETV4-overexpressing SW837 / KIR and MIA PaCa2 / KIR drug-resistant cells was extracted and the DUSP6 protein level was detected. The detection antibody used was from Abcam, catalog number ab76310. The steps of protein extraction, quantification, polyacrylamide gel electrophoresis, membrane transfer, blocking, antibody incubation, protein development, etc. were the same as above.

[0036] Test results such as Figure 4 As shown: Compared with blank SW837 / KIR and LIM2099 / KIR cells, the expression level of DUSP6 in SW837 / KIR and LIM2099 / KIR cells overexpressing ETV4 was significantly upregulated. This result suggests that the inhibition of the ETV4 / DUSP6 signaling axis is one of the main factors for ERK1 / 2 reactivation.

[0037] Through differential signal pathway enrichment analysis, the inventors further found that the activation level of the PI3K / AKT / mTOR signal pathway was significantly upregulated in drug-resistant cells. The inventors then used sotorasib to stimulate SW837, LIM2099 and MIA PaCa2 cells in the logarithmic growth phase for 0, 12, 24, 48 and 72 hours (the stimulation concentration of SW837 and MIA PaCa2 was 100nM, and the stimulation concentration of LIM2099 was 1000nM). After the treatment, each group of proteins was extracted for polyacrylamide gel electrophoresis (the detailed steps are the same as Example 2) to detect the inhibition and reactivation of the PI3K / AKT / mTOR signal pathway. The test results are as follows Figure 5 As shown: As the drug stimulation time increases, the 473rd site of AKT kinase is reactivated first.

[0038] To confirm the biological significance of the above signal, the inventors performed the following operations: S1) SW837, LIM2099 and MIA PaCa2 cells in the logarithmic growth phase were seeded in 6-well plates (seeding density 30%-40%), and after they adhered to the plate, the cells were stimulated with PI3K inhibitor LY294002 (effective concentration 10-20uM), AKT allosteric inhibitor MK2206 (effective concentration 5-10uM), AKT kinase inhibitor Capivasertib (effective concentration 10uM), mTOR1 / 2 inhibitor Torin1 (effective concentration 100nM or 1000nM) and EGFR inhibitor cetuximab (effective concentration 50ug / ml) for 72 hours; S2) After the treatment, the culture medium was discarded and the cells were washed twice with PBS. RNA was then extracted using an RNA rapid extraction kit (Yibin Biotechnology, Catalog No. RN001). The obtained RNA was quantified using a Nanodrop instrument, and the total RNA was reverse transcribed into cDNA using Evo M-MLV reverse transcription reagent premix (Aikuri Biotechnology, Catalog No. AG11706); S3) SYBR Green Pro Taq HS premixed qPCR kit (including Rox) (Acori Biotechnology, Cat. No. AG11718) and ABI fluorescent quantitative PCR instrument were used for quantitative detection to evaluate the mRNA expression level of ETV4 in each group (forward primer sequence: 5'-ACAGAGCGGACTTCGCCTAC-3', SEQ ID NO.1; reverse primer sequence: 5'-GGACAACGCAGACATCATCT-3', SEQ ID NO.2). The use of the above kits and instruments were carried out according to the relevant instructions.

[0039] Test results such as Figure 6 As shown in the figure, among the above inhibitors, only the AKT allosteric inhibitor MK2206 and the mTOR1 / 2 inhibitor Torin 1 can significantly increase the expression level of ETV4. The above results confirm that phosphorylation of AKT kinase site 473 can inhibit the expression of ETV4, thereby relieving the dephosphorylation regulation of ERK1 / 2 by the ETV4 / DUSP6 axis, causing ERK1 / 2 to be reactivated or even superactivated, ultimately leading to resistance to KRAS G12C inhibitors.

[0040] Example 3: AKT allosteric inhibitor MK2206 and KRAS G12C inhibitor sotorasib have synergistic effects This example verifies the synergistic effect of the AKT allosteric inhibitor MK2206 and the KRAS G12C inhibitor sotorasib in vivo and in vitro, respectively. The specific methods are as follows: In vitro validation experiment: SW837 / KIR, LIM2099 / KIR and MIAPaCa2 / KIR cells in logarithmic growth phase were cultured at 5×10 3 The cells were seeded at a density of 100 μg / well in a 96-well plate, and the corresponding drugs were added for treatment for 72 hours after the cells adhered to the plate. Three replicate wells were set for each drug concentration combination. After the treatment time was over, the microplate reader readings of each well were detected and the cell inhibition rate of each group was analyzed (the specific operation was the same as in Example 1), and then the ZIP model in the open source tool Synergy Finder 3.0 (https: / / synergyfinder.fimm.fi) was used to calculate the drug synergy index.

[0041] The results are as follows Figure 7 As shown: MK2206 has a high synergistic effect with sotorasib, and the ZIP scores in the SW837 / KIR, MIAPaCa2 / KIR and LIM2099 / KIR resistance models are 29.356, 29.466 and 16.729, respectively, which are significantly higher than the synergistic scores of AKT kinase inhibitor capivasertib and sotorasib.

[0042] In vivo validation experiment: We selected 48 4-6 week old balb / c-nu / nu mice, and inoculated the drug-resistant cells SW837 / KIR into the mice by subcutaneous injection, with an inoculation of 2.5 million cells per mouse. Then the mice were randomly divided into 6 groups: control group, sotorasib monotherapy group, cetuximab monotherapy group, MK2206 monotherapy group, sotorasib and cetuximab combination therapy group, and sotorasib and MK2206 combination therapy group. After the mice were routinely raised for 10 days, drug treatment began (sotorasib was administered by gavage at 100 mg / kg once a day; cetuximab was administered by tail vein injection at 50 mg / kg twice a week; MK2206 was administered by gavage at 120 mg / kg every other day), and the long and short diameters of the tumors were recorded every three days and the tumor volume was calculated. The calculation method was: tumor volume = 0.523 × long diameter × short diameter × short diameter. The drug was administered continuously for 2 weeks, and then samples were taken to weigh the tumor weight of each group and the volume growth curve was drawn.

[0043] In vivo experimental results Figure 8 As shown: Compared with the control group and other experimental groups, the tumor proliferation ability of mice in the sotorasib and MK2206 combined treatment group was significantly inhibited ( p <0.0001), while the growth rate of subcutaneous tumors in mice in the sotorasib and cetuximab combined treatment group was not statistically different from that in the control group ( p=0.8978), indicating that the combination therapy of MK2206 and sotorasib is superior to the existing clinical combination therapy and can be used as a new drug treatment combination for KRAS G12C mutant digestive system tumors.

[0044] The above is a further detailed description of the present invention, which should not be regarded as a limitation on the specific implementation of the present invention. For ordinary technicians in the technical field to which the present invention belongs, simple deduction or replacement without departing from the concept of the present invention is within the protection scope of the present invention.

Claims

1. A drug combination for treating KRAS G12C mutant digestive system tumors, characterized in that: The drug combination includes a KRAS G12C inhibitor and an AKT allosteric inhibitor.

2. The drug combination according to claim 1, characterized in that The AKT allosteric inhibitor is selected from MK2206, ARQ-092, TAS-117 or a pharmaceutically acceptable salt thereof, and its mechanism is to activate the expression of ETV4 / DUSP6 and then cooperate with the KRAS G12C inhibitor to dephosphorylate the ERK1 / 2 protein.

3. The drug combination according to claim 2, characterized in that The AKT allosteric inhibitor is selected from MK2206 or a pharmaceutically acceptable salt thereof.

4. The pharmaceutical combination according to claim 1, characterized in that The KRAS G12C inhibitor is selected from any one of sotorasib, adagrasib or fluazifop.

5. The pharmaceutical combination according to claim 1, characterized in that The KRAS G12C mutation digestive system tumor diseases include colorectal cancer and pancreatic cancer.

6. The pharmaceutical combination according to any one of claims 1 to 5, characterized in that: The drug combination further comprises pharmaceutically acceptable excipients, which include at least one of solvents, fillers, lubricants, disintegrants, buffers, solubilizers, antioxidants, antibacterial agents, emulsifiers, binders or suspending agents.

7. The pharmaceutical combination according to any one of claims 1 to 5, characterized in that: The dosage form of the drug combination is intravenous injection, suppository, gel, foam, enteric solvent, decoction, mixture, syrup, granule, pill, capsule, gel or nano preparation.

8. Use of a drug combination containing a KRAS G12C inhibitor and an AKT allosteric inhibitor in the preparation of a drug for inhibiting KRAS G12C mutant digestive system tumors.

9. The use according to claim 8, characterized in that: The KRAS G12C inhibitor is selected from any one of sotorasib, adagrasib or fluazifop, and the AKT allosteric inhibitor is selected from MK2206 or a pharmaceutically acceptable salt thereof.

10. The use according to claim 8 or 9, characterized in that: The KRAS G12C mutation digestive system tumor diseases include colorectal cancer and pancreatic cancer.

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