Pharmaceutical composition for treating lung adenocarcinoma with ANXA6 + EGFR mutation

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to application of a pharmaceutical composition in treating lung adenocarcinoma with ANXA6 + EGFR mutation. In a patient with EGFR mutation of ANXA6 + CAFs, a good treatment effect can be achieved through combined use of an EGFR-TKI targeting drug and an anti-angiogenesis drug, and in an HCC827-3 cell which is induced by ANXA6 + CAFs and tolerates AZD9291, a better treatment effect can be achieved through combined use of bevacizumab and vomitinib compared with single drug treatment. The drug combination can delay the resistance of a patient to a targeted drug, and provides accurate and effective treatment for the lung adenocarcinoma patient.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology and specifically relates to a pharmaceutical composition for treating ANXA6. + Application in EGFR-mutated lung adenocarcinoma. Background Art

[0002] Lung cancer is a malignant tumor originating from the bronchial mucosal epithelium or alveolar epithelium. It is the solid tumor with the highest morbidity and mortality in the world. In recent years, the emergence of targeted therapy, immunotherapy and anti-angiogenic drugs has improved the five-year survival rate of lung cancer patients (Chinese Medical Association Pathology Branch, National Pathology Quality Control Center, Chinese Medical Association Oncology Branch Lung Cancer Group, etc. Chinese Expert Consensus on Clinical Detection of MET in Non-Small Cell Lung Cancer [J]. Chinese Journal of Pathology, 2022, 51(11): 1094-1103.), but due to individual differences, different patients respond differently to treatment, so it is crucial to individualize the treatment method.

[0003] Lung adenocarcinoma is the most common type of lung cancer, and its diagnosis and treatment have always been the focus of medical research. According to the 2011 multidisciplinary classification of the International Association for the Study of Lung Cancer (IASLC), the American Thoracic Society (ATS), and the European Respiratory Society (ERS), the classification of lung adenocarcinoma has been updated to better reflect its biological characteristics and clinical behavior (International Association for the Study of Lung Cancer / American Thoracic Society / European Respiratory Society International Multidisciplinary Classification of Lung Adenocarcinoma Travis, William D. et al. Journal of Thoracic Oncology, Volume 6, Issue 2, 244-285). In terms of treatment, molecular targeted therapy has become an important treatment for advanced lung adenocarcinoma. For example, patients with epidermal growth factor receptor (EGFR) mutations can benefit from EGFR tyrosine kinase inhibitors (Wang Cai, Wei Suju, Hong Lei, et al. Analysis of clinical prognostic factors in 319 patients with advanced lung adenocarcinoma [J]. Oncology Basics and Clinics, 2016, 29(03): 240-246.). In addition, chemotherapy regimens such as pemetrexed combined with cisplatin have also shown efficacy in patients with advanced lung adenocarcinoma ([1] Chen Hua, Xu Haixia, Hu Qin. Clinical efficacy of different regimens in the treatment of advanced lung adenocarcinoma and their effects on tumor marker levels [J]. Journal of Clinical Military Medicine, 2020, 48(09): 1066-1068.).

[0004] Currently, about 50% of Asian lung adenocarcinoma patients have EGFR-sensitive mutations. A variety of targeted epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) have been approved for the treatment of patients with EGFR gene-sensitive mutations, which benefit patients. In particular, the third-generation EGFR-TKIs have significantly prolonged patients' survival and improved their quality of life. However, drug resistance is inevitable in the end, which greatly reduces the efficacy.

[0005] Bevacizumab is a recombinant humanized immunoglobulin G1 monoclonal antibody that inhibits the activity of vascular endothelial growth factor (VEGF) and angiogenesis, blocks the binding of VEGF to receptors, reduces the formation of new blood vessels in the tumor site, and achieves the purpose of inhibiting tumor growth. It can be used to treat a variety of tumors such as colorectal cancer, lung cancer, and ovarian cancer (Zhou Zhenxing, Song Junmin, Chen Jihua, et al. Research progress on the application of bevacizumab in tumor treatment [J]. Pharmaceutical Progress, 2015, 39(7): 525-532.). However, in clinical practice, the use of bevacizumab alone has certain limitations. The most commonly used method now is to use it in combination with a variety of chemotherapy drugs to achieve the purpose of better inhibiting tumor progression.

[0006] ANXA6 + Fibroblasts (ANXA6 + CAFs can cause resistance to EGFR-TKI-sensitive lung adenocarcinoma cells, and this resistance is related to ANXA6 + CAFs are closely related to the promotion of angiogenesis in lung cancer. + In patients with EGFR mutations in CAFs, the combined use of EGFR-TKI targeted drugs and anti-angiogenic drugs can achieve good therapeutic effects and delay the patient's resistance to targeted drugs.

[0007] In short, the treatment of lung adenocarcinoma requires a personalized treatment plan based on the patient's specific pathological type, molecular characteristics, and clinical condition. With the development of molecular biology and precision medicine, more effective treatment options are expected to be provided for lung adenocarcinoma patients in the future. Summary of the invention

[0008] The present invention discovered that ANXA6 + Fibroblasts (ANXA6 + The present invention is based on the fact that CAFs can lead to drug resistance in lung adenocarcinoma cells that are sensitive to EGFR-TKI. The combined use of EGFR-TKI targeted drugs and anti-angiogenic drugs can achieve good therapeutic effects and delay the resistance of patients to targeted drugs.

[0009] In a first aspect, the present invention provides a method for treating ANXA6 +A pharmaceutical composition for lung adenocarcinoma resistant to EGFR-TKI caused by CAFs, the pharmaceutical composition comprising an EGFR-TKI targeted drug and an anti-tumor angiogenesis drug, and the pharmaceutical composition has at least one of the following effects:

[0010] Inhibit angiogenesis in lung cancer;

[0011] Reduce tumor volume;

[0012] Preventing and / or treating lung adenocarcinoma.

[0013] Furthermore, the EGFR-TKI targeted drugs include gefitinib, erlotinib, icotinib, afatinib, dacomitinib, osimertinib, ametinib, vometinib and befortinib, preferably vometinib and / or osimertinib.

[0014] Furthermore, the gefitinib, erlotinib, icotinib, afatinib, dacomitinib, osimertinib, ametinib, vometinib and befortinib also include pharmaceutically acceptable salts or esters thereof.

[0015] Furthermore, the anti-tumor angiogenesis drug is bevacizumab.

[0016] Furthermore, one or more pharmaceutically acceptable carriers may be added to the pharmaceutical composition.

[0017] Furthermore, the pharmaceutical composition can be prepared into various forms such as injection, tablet, powder, granule, capsule, oral solution, injection preparation or aerosol; the above various dosage forms of drugs can be prepared according to conventional methods in the pharmaceutical field.

[0018] Furthermore, the preparation can be one or more of a conventional preparation, a sustained-release preparation, a controlled-release preparation and / or various microparticle delivery systems.

[0019] Furthermore, the tablets may widely use various carriers known in the art, including one or more of diluents and absorbents, wetting agents and binders, disintegrants, disintegration inhibitors, absorption promoters and / or lubricants.

[0020] Furthermore, the diluent and absorbent include, but are not limited to, one or more of starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose and / or aluminum silicate.

[0021] Furthermore, the wetting agent and adhesive include, but are not limited to, one or more of water, glycerol, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, acacia gum slurry, gelatin slurry, sodium carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate and / or polyvinyl pyrrolidone.

[0022] Furthermore, the disintegrant includes, but is not limited to, one or more of dry starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene, sorbitan fatty acid ester, sodium dodecyl sulfate, methyl cellulose and / or ethyl cellulose.

[0023] Furthermore, the disintegration inhibitor includes but is not limited to sucrose, tristearin, cocoa butter and / or hydrogenated oil.

[0024] Furthermore, the absorption enhancer includes but is not limited to one or more of quaternary ammonium salts and / or sodium lauryl sulfate.

[0025] Furthermore, the lubricant includes, but is not limited to, one or more of talc, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin and / or polyethylene glycol.

[0026] Furthermore, the tablets can be further made into coated tablets, including sugar-coated tablets, film-coated tablets, enteric-coated tablets, double-layer tablets and multi-layer tablets.

[0027] Furthermore, the injectable preparation includes, but is not limited to, one or more of a solution, an emulsion, a lyophilized powder injection and / or a suspension.

[0028] Furthermore, the injectable preparation may use all diluents commonly used in the art, including but not limited to one or more of water, ethanol, polyethylene glycol, 1,3-propylene glycol, ethoxylated isostearyl alcohol, polyoxygenated isostearyl alcohol and / or polyoxyethylene sorbitan fatty acid esters.

[0029] Furthermore, in order to prepare an isotonic injection solution, the injection preparation may be added with an appropriate amount of one or more of sodium chloride, glucose, glycerol, conventional cosolvents, buffers and / or pH regulators.

[0030] Furthermore, the various preparations may also add colorants, preservatives, spices, flavoring agents, sweeteners or other materials to the pharmaceutical preparations as needed.

[0031] Furthermore, the pharmaceutical composition can be introduced into the body by physical or chemical methods, such as intramuscularly, intradermally, subcutaneously or intravenously.

[0032] In a second aspect, the present invention provides a method for treating ANXA6 by combining an EGFR-TKI targeted drug and an anti-tumor angiogenesis drug. + Use of CAFs in a pharmaceutical composition for treating EGFR-TKI-resistant lung adenocarcinoma.

[0033] Furthermore, the EGFR-TKI targeted drugs include gefitinib, erlotinib, icotinib, afatinib, dacomitinib, osimertinib, ametinib, vometinib and befortinib, preferably vometinib and / or osimertinib.

[0034] Furthermore, the gefitinib, erlotinib, icotinib, afatinib, dacomitinib, osimertinib, ametinib, vometinib and befortinib also include pharmaceutically acceptable salts or esters thereof.

[0035] Furthermore, the anti-tumor angiogenesis drug is bevacizumab.

[0036] Furthermore, one or more pharmaceutically acceptable carriers may be added to the pharmaceutical composition.

[0037] Furthermore, the pharmaceutical composition can be prepared into various forms such as injection, tablet, powder, granule, capsule, oral solution, injection preparation or aerosol; the above various dosage forms of drugs can be prepared according to conventional methods in the pharmaceutical field.

[0038] Furthermore, the preparation can be one or more of a conventional preparation, a sustained-release preparation, a controlled-release preparation and / or various microparticle delivery systems.

[0039] Furthermore, the tablets may widely use various carriers known in the art, including one or more of diluents and absorbents, wetting agents and binders, disintegrants, disintegration inhibitors, absorption promoters and / or lubricants.

[0040] Furthermore, the diluent and absorbent include, but are not limited to, one or more of starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose and / or aluminum silicate.

[0041] Furthermore, the wetting agent and adhesive include, but are not limited to, one or more of water, glycerol, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, acacia gum slurry, gelatin slurry, sodium carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate and / or polyvinyl pyrrolidone.

[0042] Furthermore, the disintegrant includes, but is not limited to, one or more of dry starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene, sorbitan fatty acid ester, sodium dodecyl sulfate, methyl cellulose and / or ethyl cellulose.

[0043] Furthermore, the disintegration inhibitor includes but is not limited to sucrose, tristearin, cocoa butter and / or hydrogenated oil.

[0044] Furthermore, the absorption enhancer includes but is not limited to one or more of quaternary ammonium salts and / or sodium lauryl sulfate.

[0045] Furthermore, the lubricant includes, but is not limited to, one or more of talc, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin and / or polyethylene glycol.

[0046] Furthermore, the tablets can be further made into coated tablets, including sugar-coated tablets, film-coated tablets, enteric-coated tablets, double-layer tablets and multi-layer tablets.

[0047] Furthermore, the injectable preparation includes, but is not limited to, one or more of a solution, an emulsion, a lyophilized powder injection and / or a suspension.

[0048] Furthermore, the injectable preparation may use all diluents commonly used in the art, including but not limited to one or more of water, ethanol, polyethylene glycol, 1,3-propylene glycol, ethoxylated isostearyl alcohol, polyoxygenated isostearyl alcohol and / or polyoxyethylene sorbitan fatty acid esters.

[0049] Furthermore, in order to prepare an isotonic injection solution, the injection preparation may be added with an appropriate amount of one or more of sodium chloride, glucose, glycerol, conventional cosolvents, buffers and / or pH regulators.

[0050] Furthermore, the various preparations may also add colorants, preservatives, spices, flavoring agents, sweeteners or other materials to the pharmaceutical preparations as needed.

[0051] Furthermore, the pharmaceutical composition can be introduced into the body by physical or chemical methods, such as intramuscularly, intradermally, subcutaneously or intravenously.

[0052] Beneficial Effects

[0053] In ANXA6 + In patients with EGFR mutations in CAFs, the combined use of EGFR-TKI targeted drugs and anti-angiogenic drugs can achieve good therapeutic effects and delay patients' resistance to targeted drugs, providing precise and effective treatment for patients with lung adenocarcinoma. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 ANXA6 + CAFs promote angiogenesis and cell resistance in EGFR-mutated lung adenocarcinoma cells.

[0055] Note: A: ANXA6 + CAFs cell supernatant treated HCC827 cells, HCC827 cells response to AZD9291; B: ANXA6 +PC-9 cells treated with CAFs cell supernatant, and the response of PC-9 cells to AZD9291; C: ANXA6 + Angiogenesis in CAFs+HCC827 group; D: ANXA6 + Angiogenesis in the CAFs+PC-9 group.

[0056] Figure 2 In ANXA6 + CAFs induce the effect of bevacizumab combined with vometinib in EGFR-TKI-resistant cells. Note: A: ANXA6 + Response of HCC827 cells to bevacizumab combined with vometinib in CAFs+HCC827 transplanted tumors; B: ANXA6 + Response of PC-9 cells to bevacizumab combined with vometinib in CAFs+PC-9 transplanted tumors.

[0057] Figure 3 In ANXA6 + CAFs induce the therapeutic effect of bevacizumab combined with vometinib in EGFR-TKI-resistant cells. Note: A: ANXA6 + Response of HCC827 cells to bevacizumab combined with vometinib in CAFs+HCC827 transplanted tumors; B: ANXA6 + Response of PC-9 cells to bevacizumab combined with vometinib in CAFs+PC-9 transplanted tumors.

[0058] Figure 4 The therapeutic effect of bevacizumab combined with vometinib in EGFR-TKI-resistant cell lines.

[0059] Figure 5 Effect of combined treatment of bevacizumab and vometinib on HCC827-3 cells. DETAILED DESCRIPTION

[0060] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in the embodiments described below can be combined with each other as long as they do not conflict with each other.

[0061] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0062] Test materials

[0063] The basal culture medium used in the example is: serum-free RPMI-1640 / DMEM culture medium, fetal bovine serum is added to a concentration of 10%, penicillin is added to a concentration of 1%, and streptomycin is added to a concentration of 1%.

[0064] Example 1 ANXA6 + CAFs promote angiogenesis in HCC827 / PC-9 cell transplanted tumors

[0065] (1) HCC827 / PC-9 cells and ANXA6 + CAFs cells were digested into single cells and counted;

[0066] (2) HCC827 / PC-9 cell suspension and ANXA6 + CAFs cell suspension was mixed at a ratio of 1:1;

[0067] (3) Prepare 4- to 6-week-old BALB / c nude mice and inoculate the cell suspension into the axilla of the nude mice, injecting 2 million cells in each site;

[0068] (4) When nude mice have subcutaneous tumors, the mice are killed, and the transplanted tumors are dissected and fixed;

[0069] (5) HE staining observation and record.

[0070] Example 2 Immunohistochemistry

[0071] (1) Tissue processing: slice the tissue to be examined and dry it to ensure that the immunohistochemical reagent can bind to the tissue;

[0072] (2) Antigen repair: heating or enzymatic hydrolysis, etc.

[0073] (3) Blocking nonspecific binding: adding nonspecific antiserum, protein solution or other blocking reagents;

[0074] (4) Primary antibody incubation: CD31 primary antibody was coated on the tissue sections and incubated;

[0075] (5) Secondary antibody incubation: Apply the labeled secondary antibody to the tissue section and incubate;

[0076] (6) Colorimetric observation: Observe the distribution and expression of the target antigen in the tissue.

[0077] like Figure 1 As shown in C and 1D, compared with the HCC827 / PC-9 group, ANXA6 + CAFs+HCC827 / ANXA6 +The tumor angiogenesis in the CAFs+PC-9 group was significantly increased. The detection of endothelial cell marker CD31 confirmed that compared with the HCC827 / PC-9 group, ANXA6 + CAFs+HCC827 / ANXA6 + The tumor angiogenesis in the CAFs+PC-9 group was significantly increased.

[0078] Example 3 Using ANXA6 + Treatment of lung cancer cells with CAFs supernatant and drug sensitivity test

[0079] (1)ANXA6 + CAFs cells were cultured in DMEM medium, and the cell culture supernatant was collected;

[0080] (2) HCC827 and PC-9 cells were inoculated in 10% RPMI-1640 medium, passaged, digested into single cells, and counted;

[0081] (3) Inoculate HCC827 and PC-9 cells in well plates;

[0082] (4) ANXA6 + CAFs cell culture supernatant was mixed with 10% RPMI-1640 medium at a volume ratio of 1:1, and different concentrations of AZD9291, including 6 nM, 12 nM, 120 nM, 250 nM, 500 nM, 1200 nM, 6000 nM, 12000 nM, and 24000 nM, were prepared and added to the well plate inoculated with HCC827 cells;

[0083] (5) ANXA6 + The CAFs cell culture supernatant was mixed with 10% RPMI-1640 medium at a volume ratio of 1:1, and different concentrations of AZD9291, including 1 nM, 2 nM, 4 nM, 40 nM, 400 nM, 4000 nM, 6000 nM, 8000 nM, and 16000 nM, were prepared and added to the well plate inoculated with PC-9 cells;

[0084] (6) Draw the drug sensitivity curve of cells to AZD9291 based on the data.

[0085] like Figure 1 As shown in A and 1B, ANXA6 + CAFs promote angiogenesis and cell resistance in EGFR-mutated lung adenocarcinoma cells. + After HCC827 and PC-9 cells were treated with CAFs cell supernatant, the cells became resistant to AZD9291.

[0086] Example 4 Obtaining AZD9291-tolerant cells HCC827-3

[0087] A. Obtaining cells

[0088] (1) HCC827 / PC-9 cells and ANXA6 + CAFs cells were digested into single cells and counted;

[0089] (2) HCC827 / PC-9 cell suspension and ANXA6 + CAFs cell suspension was mixed at a ratio of 1:1;

[0090] (3) Prepare 4- to 6-week-old BALB / c nude mice and inoculate the cell suspension into the axilla of the nude mice, injecting 2 million cells in each site;

[0091] (4) After nude mice have subcutaneous tumors, the mice are killed and the tumors are dissected out;

[0092] (5) Cut the tumor into pieces, wash them, add DMEM medium and culture them;

[0093] (6) After the tumor cells adhered to the wall and grew, they were digested, subcultured and named HCC827-3 cells.

[0094] B. Verify tolerance

[0095] (1) Inoculate HCC827 and HCC827-3 cells in a well plate;

[0096] (2) AZD9291 at different concentrations of 0 nM, 3 nM, 6 nM, 12 nM, 120 nM, 250 nM, 500 nM, 1200 nM, 6000 nM, 12000 nM, and 24000 nM was prepared and added to the well plate inoculated with HCC827 and HCC827-3 cells;

[0097] (3) Draw the drug sensitivity curves of the two cell types to AZD9291 based on the data;

[0098] (4) HCC827 and HCC827-3 were injected into lung metastatic lesions through the tail vein and treated with vometinib to observe their response to the drug.

[0099] C. Test Results

[0100] like Figure 2 As shown in A and 2B, the IC50 of HCC827 cells to AZD9291 is 11.75 nM, and the IC50 of HCC827-3 cells to AZD9291 is 10232 nM, which is 870.81 times that of HCC827 cells. HCC827-3 cells are resistant to AZD9291.

[0101] like Figure 2 As shown in C and 2D, the therapeutic effect of vometinib on lung metastases formed by HCC827 cells was significantly better than that on lung metastases formed by HCC827-3 cells. These results showed that HCC827-3 cells were resistant to the third-generation EGFR-TKI.

[0102] Example 5 Bevacizumab and Vometinib in the treatment of ANXA6 + Cancer cells treated with CAFs

[0103] (1) HCC827 / PC-9 cells and ANXA6 + CAFs cells were digested into single cells and counted.

[0104] Cell suspension and ANXA6 + CAFs cell suspension was mixed at a ratio of 1:1;

[0105] (2) Prepare 4- to 6-week-old BALB / c nude mice and inoculate the cell suspension into the axilla of the nude mice, injecting 2 million cells in each site;

[0106] (3) When the tumor grows to 500 mm 3 The patients were divided into control group, bevacizumab treatment group, vometinib treatment group and bevacizumab and vometinib treatment group. Bevacizumab was intraperitoneally injected at 15 mg / kg twice a week, and vometinib was intragastrically injected at 5 mg / kg.

[0107] (4) For large tumors 1000 mm 3 , divided into a vometinib treatment group and a bevacizumab and vometinib treatment group, with the same dosage of bevacizumab and vometinib (3).

[0108] (5) Observe the tumor formation in the axilla of nude mice, measure the tumor volume, and draw the growth curve of the transplanted tumor in each group.

[0109] like Figure 3 As shown, in ANXA6 + Smaller CAFs+HCC827 / PC-9 xenografts (volume 500 mm 3 ), bevacizumab combined with vometinib had a good therapeutic effect compared with the control and single drug.

[0110] like Figure 4 As shown, in ANXA6 + CAFs+HCC827 / PC-9 larger xenografts (volume 1000mm 3 ), bevacizumab combined with vometinib had a good therapeutic effect compared with the control and single drug.

[0111] Example 6 Combination therapy of bevacizumab and vometinib in HCC827-3 cells

[0112] (1) Prepare HCC827-3 cells into a single cell suspension of 10 million / ml;

[0113] (2) Prepare 4- to 6-week-old BALB / c nude mice and inoculate the cell suspension into the axilla of the nude mice, injecting 2 million cells in each site;

[0114] (3) For large tumors 1000 mm 3 The patients were divided into a vumetinib treatment group and a bevacizumab and vumetinib treatment group. Bevacizumab was intraperitoneally injected at 15 mg / kg twice a week, and vumetinib was intragastrically injected at 5 mg / kg.

[0115] (4) Observe the tumor condition in the axilla of nude mice, measure the tumor volume, and draw the growth curve of the transplanted tumor in each group.

[0116] like Figure 5 As shown, in HCC827-3 cells that tolerate AZD9291, bevacizumab combined with vometinib treatment for 6 days significantly reduced the tumor, and the treatment effect could be stabilized at a good level for a long time; although the tumor reduction rate of vometinib alone was not as fast as that of the double-drug group, the tumor was also significantly reduced after 6 days of treatment. The difference was that the good treatment effect of the single drug only lasted for 6 days. After 12 days of treatment, the tumor began to show a slow increase trend. These results showed that ANXA6 + In CAFs-induced AZD9291-resistant HCC827-3 cells, the combined use of bevacizumab and vometinib can achieve better therapeutic effects.

Claims

1. A treatment for ANXA6 + A pharmaceutical composition for lung adenocarcinoma resistant to EGFR-TKI caused by CAFs, the pharmaceutical composition comprising an EGFR-TKI targeted drug and an anti-tumor angiogenesis drug, and the pharmaceutical composition has at least one of the following effects: Inhibit angiogenesis in lung cancer; Reduce tumor volume; Preventing and / or treating lung adenocarcinoma.

2. A combination of EGFR-TKI targeted drug and anti-angiogenic drug in the treatment of ANXA6 + Use of CAFs in a pharmaceutical composition for treating EGFR-TKI-resistant lung adenocarcinoma.

3. The EGFR-TKI targeted drug according to any one of claims 1 or 2 comprises gefitinib, erlotinib, icotinib, afatinib, dacomitinib, osimertinib, ametinib, vometinib, befortinib and pharmaceutically acceptable salts or esters thereof, preferably vometinib and / or osimertinib.

4. The anti-tumor angiogenesis drug according to any one of claims 1 or 2 is bevacizumab.

5. The pharmaceutical composition according to any one of claims 1 or 2, further comprising one or more pharmaceutically acceptable carriers.

6. The pharmaceutical composition according to any one of claims 1 or 2, which can be introduced into the body by physical or chemical methods, such as intramuscularly, intradermally, subcutaneously or intravenously.

7. The pharmaceutical composition according to any one of claims 1 or 2 can be prepared into various forms such as injection, tablets, powders, granules, capsules, oral solutions, injection preparations or aerosols; the above-mentioned various dosage forms of drugs can be prepared according to conventional methods in the pharmaceutical field.

8. The preparation as claimed in claim 7 can be one or more of a conventional preparation, a sustained-release preparation, a controlled-release preparation and / or various microparticle delivery systems.

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