Use of pyruvate in the preparation of a drug for enhancing the efficacy of lung cancer chemotherapy

Abstract

The present invention belongs to the field of biomedical technologies, and particularly relates to the application of pyruvate in the preparation of a drug for enhancing the efficacy of lung cancer chemotherapy. The present invention provides a novel drug for treating lung cancer, namely Compound I obtained by structurally modifying sodium pyruvate. Experimental results show that this compound can significantly inhibit the proliferation of lung cancer cells A549 in mice, and at the same time has an inhibitory effect on the growth of human lung cancer A549 cell xenografts in nude mice, expanding the therapeutic potential of existing drugs and providing new treatment options for lung cancer patients. Its high efficiency in treating lung cancer is of great significance in improving the treatment effect and reducing the burden on patients.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to the application of pyruvate in the preparation of a drug for enhancing the efficacy of lung cancer chemotherapy. Background Art

[0002] With the rapid development of the urbanization process and the increasing aging of the population, the morbidity and mortality of malignant tumors have been continuously increasing globally. Lung cancer is the leading cause of cancer death worldwide and seriously endangers human health.

[0003] Lung cancer is mainly divided into two histological types: non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). NSCLC accounts for about 85%, and SCLC accounts for about 15%. The cancer cells of NSCLC grow and divide relatively slowly, and the spread and metastasis are relatively late. NSCLC can be further subdivided into multiple subtypes such as adenocarcinoma, squamous cell carcinoma, adenosquamous carcinoma, large cell carcinoma, and sarcomatoid carcinoma, mainly presenting local symptoms in the lungs, such as cough, hemoptysis, chest pain, and low fever. According to different stages, various treatment methods such as surgery, radiotherapy, chemotherapy, targeted therapy, and immunotherapy can be applied. If detected and treated early in a timely manner, the prognosis is relatively good. The cancer cells of SCLC are smaller, shaped like oat grains, grow rapidly, are highly invasive, metastasize distantly early, have a high degree of malignancy, and are mostly related to smoking. In addition to lung symptoms, obvious systemic symptoms may also appear, such as paraneoplastic syndromes like syndrome of inappropriate antidiuretic hormone secretion and Cushing's syndrome. Due to rapid growth and easy metastasis, it is mostly sensitive to radiotherapy and chemotherapy. However, metastasis often has occurred at the time of diagnosis, and the effect of surgical treatment is limited. Therefore, the preferred treatment is comprehensive radiotherapy and chemotherapy. Obviously, radiotherapy and chemotherapy are effective treatment methods for NSCLC and SCLC.

[0004] There are a wide variety of chemical drugs used for lung cancer chemotherapy, such as cisplatin injection, paclitaxel injection, carboplatin injection, gemcitabine, pemetrexed disodium, ifosfamide, doxorubicin, vincristine, vinblastine, and etoposide, which inhibit or kill lung cancer cells through different mechanisms. However, the extensive use of these drugs has relatively large side effects. For example, the toxicity to the kidneys and gastrointestinal reactions may cause damage to renal tubules, elevated blood urea nitrogen and serum creatinine levels; poor water solubility, and traditional dosage forms need to use hydrogenated castor oil and absolute ethanol as carriers, which may cause adverse reactions such as allergies, nephrotoxicity, and neurotoxicity; allergic reactions are likely to occur during multiple applications, and hypotensive shock may occur in severe cases; the cardiotoxicity is significant, and serious side effects such as abnormal electrocardiogram and arrhythmia may be caused.

[0005] Maximum tolerated dose chemotherapy (MTD) is the main form of traditional chemotherapy. Due to its relatively severe toxic and side effects, an intermission of 2-3 weeks is required after each cycle of chemotherapy to facilitate the recovery of the functions of the body's tissues and organs, and it is easy to produce tumor drug resistance. In recent years, adjuvant chemoradiotherapy sensitizing drugs have been mostly used in clinical tumor treatment. The purpose is to maximize the killing effect and reduce the incidence of tumor drug resistance without increasing the toxic effects. However, these drugs are expensive and have high drug adverse reactions after long-term use, resulting in poor patient compliance. Therefore, searching for and developing new, highly effective and low-toxic lung cancer chemotherapeutic agents and reducing the chemotherapy dosage have important practical significance for the treatment of lung cancer and the reversal / delay of lung cancer drug resistance. Summary of the Invention

[0006] In order to overcome the deficiencies of the prior art, one of the purposes of the present invention is to provide the application of pyruvate in the preparation of a lung cancer chemotherapy synergistic drug, which can ensure the chemotherapy effect while reducing the chemotherapy dosage.

[0007] One of the purposes of the present invention is realized by adopting the following technical scheme:

[0008] The application of pyruvate in the preparation of a lung cancer chemotherapy synergistic drug, and the structural formula of the pyruvate is shown as follows:

[0009] .

[0010] Further, the pyruvate is Compound I, and the preparation process of the Compound I includes the following steps:

[0011]

[0012] (1) Sodium pyruvate is added to the acetic acid solution of 4-bromo-o-phenylenediamine, and after reacting for 3-5 h, Intermediate 2A is obtained through treatment;

[0013] (2) 4-Methoxy-α-bromoacetophenone, thiosemicarbazide, and an inorganic base are mixed evenly, and after microwave reaction at 190-205 °C for 5-10 min, Intermediate 1 is obtained through treatment;

[0014] (3) The Intermediate 1 and 4-isothiocyanatophenol are successively added to a solvent, and after reacting under reflux for 2-3 h, Intermediate 2 is obtained through treatment;

[0015] (4) The Intermediate 2, the Intermediate 2A, an inorganic base, and copper powder are added to a solvent, and after microwave reaction at 150-170 °C for 25-35 min, Compound I is obtained through treatment.

[0016] Further, the dosage ratio of sodium pyruvate, 4-bromo-o-phenylenediamine, and acetic acid in step (1) is 1 mmol : (1 - 1.25) mmol : (2 - 2.5) mL.

[0017] Further, the molar dosage ratio of 4-methoxy-α-bromoacetophenone, thiosemicarbazide, and inorganic base in step (2) is 1 : (1 - 1.5) : (9 - 11).

[0018] Further, the molar dosage ratio of intermediate 1 and 4-isothiocyanatophenol in step (3) is 1 : (1 - 1.25).

[0019] Further, the molar dosage ratio of intermediate 2, intermediate 2A, inorganic base, and copper powder in step (4) is 1 : (1 - 1.25) : (2 - 3) : (0.1 - 0.25).

[0020] Further, the inorganic base in steps (2) and (4) is one of cesium carbonate and potassium carbonate.

[0021] Further, the solvent in step (3) is one of toluene and xylene, and the solvent in step (4) is one of DMF and DMAC.

[0022] Further, the compound I is used to prepare a drug for treating lung cancer.

[0023] Further, the drug also includes pharmaceutically acceptable excipients.

[0024] Furthermore, the power of the microwave reaction in steps (2) and (4) is 800 W, and the frequency is 2500 Hz.

[0025] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0026] By chemically modifying the pyruvate molecule, the present invention obtains a derivative compound I of pyruvate with higher anti-cancer activity. This compound can inhibit half of the A549 lung cancer cells at a lower concentration (0.287 μM), and it can inhibit the epithelial-mesenchymal transition (EMT) process. At the same time, it has an inhibitory effect on the growth of human lung cancer A549 cell xenografts in nude mice. Description of the Drawings

[0027] Figure 1 It is the synthesis route of compound I of the present invention;

[0028] Figure 2 It is the schematic diagram of the cell effect of compound I of the present invention on A549 lung cancer cells;

[0029] Figure 3Schematic diagram of the effect of compound Ⅰ of the present invention on E-cadherin in the EMT process of cells;

[0030] Figure 4 Schematic diagram of the effect of compound Ⅰ of the present invention on Vimentin in the EMT process of cells;

[0031] Figure 5 Schematic diagram of the effect of compound Ⅰ of the present invention on Fibronectin in the EMT process of cells;

[0032] Figure 6 Schematic diagram of the effect of compound Ⅰ of the present invention on N-cadherin in the EMT process of cells. Detailed implementation manners

[0033] Next, in combination with the accompanying drawings and specific implementation manners, the present invention will be further described. It should be noted that on the premise of no conflict, the following-described embodiments or technical features can be arbitrarily combined with each other to form new embodiments. The specific conditions not specified in the embodiments are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used, unless otherwise specified, are all conventional products obtained through commercial channels.

[0034] Example 1

[0035] Application of pyruvate in the preparation of a chemosensitizer for lung cancer chemotherapy, wherein the pyruvate is compound Ⅰ, and the structural formula of the compound Ⅰ is as follows:

[0036] 。

[0037] The synthesis route of the compound Ⅰ is as Figure 1 shown, including the following steps:

[0038] (1) Place 4-bromo-o-phenylenediamine (20 mmol) in 50 mL of acetic acid, then dissolve sodium pyruvate (20 mmol) in 9 mL of deionized water, and drop the sodium pyruvate solution into the 4-bromo-o-phenylenediamine solution under stirring conditions, react at room temperature for 3 h, filter the reaction solution, wash the filter cake with deionized water and then dry to obtain intermediate 2A.

[0039] The 1 1H NMR (C9H7N2OBr, 400 MHz, DMSO-d6) δ: 10.68 (s, 1H), 7.54 - 7.47 (m, 3H), 2.07 (s, 3H); [M + H] + m / z: 238.97, ESI-MS (m / z): 239.30. 1 The results of 1HNMR and MS prove it to be the target product.

[0040] (2) 4-Methoxy-α-bromoacetophenone (10 mmol) and thiosemicarbazide (10 mmol) were added into a microwave reactor (800 W, 2500 HZ), then potassium carbonate (0.11 mol) was added, and the reaction was carried out at 200 °C for 5 min; after cooling to room temperature, the reaction solution was diluted with sufficient water, and then the reaction solution was filtered to obtain a filter cake. The filter cake was slurried with methanol and dried to obtain Intermediate 1.

[0041] For Intermediate 1, 1 1H NMR (C 10 1H 11 N3OS, 400 MHz, DMSO-d6) δ: 9.30 (s, 2H), 7.94 (d, 2H), 7.08 (d, 2H), 3.83 (s, 3H), 3.72 (s, 2H); [M+H] + m / z: 222.06, ESI-MS (m / z): 222.16. 1 The results of 1H NMR and MS proved it to be the target product.

[0042] (3) Intermediate 1 (20 mmol) and 4-isothiocyanatophenol (20 mmol) were added into 50 mL of toluene, and the reaction was refluxed in toluene for 2 h; after the reaction, the system was cooled to room temperature, filtered to obtain a filter cake, and the filter cake was washed successively with toluene and n-hexane, and then slurried with ethanol and dried to obtain Intermediate 2.

[0043] For Intermediate 2, 1 1H NMR (C 17 1H 16 N4O2S2, 400 MHz, DMSO-d6) δ: 12.24 (s, 1H), 10.96 (s, 1H), 9.42 (s, 1H), 7.94 (d, 2H), 7.27 (d, 2H), 7.08 (d, 2H), 6.72 (d, 2H), 3.83 (s, 3H), 3.72 (s, 2H); [M+H] + m / z: 373.07, ESI-MS (m / z): 373.12. 1 The results of 1H NMR and MS proved it to be the target product.

[0044] (4) Intermediate 2A (6 mmol) and Intermediate 2 (5 mmol) were added into 6 mL of N,N-dimethylformamide (DMF), potassium carbonate (10 mmol) and copper powder (1.25 mmol) were added as catalysts, and the reaction was carried out in a microwave reactor (800 W, 2500 HZ) at 150 °C for 30 min; after the reaction was completed, the mixture was cooled to room temperature, then the reaction was quenched with water, and the organic layer was obtained by extraction with ethyl acetate and then dried and column chromatographed to obtain Compound I.

[0045] Of Compound I 1 1H NMR (C 26 1 22 6H6O3S2, 400 MHz, DMSO-d6) δ: 12.24 (s, 1H), 10.96 (s, 1H), 10.68 (s, 1H), 7.94 (d, 2H), 7.41 (d, 2H), 7.20 - 7.16 (d, 7H), 3.83 (s, 3H), 3.72 (s, 2H), 2.07 (s, 3H); [M + H] + m / z: 531.12, ESI-MS (m / z): 531.12. 1 The results of 1H NMR and MS prove it to be the target product.

[0046] Example 2

[0047] Use of a pyruvate salt in the preparation of a drug for enhancing the efficacy of lung cancer chemotherapy, wherein the pyruvate salt is Compound I, and the preparation process of the shown Compound I includes the following steps:

[0048] (1) Place 4-bromo-o-phenylenediamine (25 mmol) in 40 mL of acetic acid, then dissolve sodium pyruvate (20 mmol) in 15 mL of deionized water, and then dropwise add sodium pyruvate to the 4-bromo-o-phenylenediamine solution under stirring. React at room temperature for 4 h, filter the reaction solution, wash the filter cake with deionized water and then dry to obtain Intermediate 2A.

[0049] (2) Add 4-methoxy-α-bromoacetophenone (10 mmol) and thiosemicarbazide (15 mmol) to a microwave reactor (800 W, 2500 HZ), then add cesium carbonate (0.1 mol), and react at 190 °C for 10 min; after cooling to room temperature, add a sufficient amount of water to dilute the reaction solution, and then filter the reaction solution to obtain a filter cake. The filter cake is slurried with methanol and then dried to obtain Intermediate 1.

[0050] (3) Add Intermediate 1 (20 mmol) and 4-isothiocyanatophenol (25 mmol) to 50 mL of xylene, and reflux in xylene for 3 h; after the reaction, cool the system to room temperature, filter to obtain a filter cake, wash the filter cake with xylene and n-hexane in sequence, and then slurry with ethanol and dry to obtain Intermediate 2. 1 The results of 1H NMR and MS are consistent with those of Example 1.

[0051] (4) Intermediate 2A (5 mmol) and Intermediate 2 (5 mmol) were added to 6 mL of N,N-dimethylacetamide (DMAC), and cesium carbonate (15 mmol) and copper powder (1 mmol) were added as catalysts. The reaction was carried out at 170 °C for 25 min in a microwave reactor (800 W, 2500 HZ). After the reaction was completed, the mixture was cooled to room temperature, and then the reaction was quenched with water. The organic layer was obtained by extraction with ethyl acetate and then dried and column chromatographed to obtain Compound I. 1 The results of HNMR and MS were consistent with those of Example 1.

[0052] Example 3

[0053] Use of a pyruvate salt in the preparation of a drug for enhancing the efficacy of lung cancer chemotherapy, wherein the pyruvate salt is Compound I, and the preparation process of the shown Compound I includes the following steps:

[0054] (1) 4-Bromo-o-phenylenediamine (23 mmol) was placed in 45 mL of acetic acid, and then sodium pyruvate (20 mmol) was dissolved in 22 mL of deionized water. Then, sodium pyruvate was added dropwise to the 4-bromo-o-phenylenediamine solution under stirring conditions, and the reaction was carried out at room temperature for 5 h. The reaction solution was filtered, and the filter cake was washed with deionized water and then dried to obtain Intermediate 2A.

[0055] (2) 4-Methoxy-α-bromoacetophenone (10 mmol) and thiosemicarbazide (12 mmol) were added to a microwave reactor (800 W, 2500 HZ), and then potassium carbonate (0.09 mol) was added. The reaction was carried out at 205 °C for 8 min. After cooling to room temperature, the reaction solution was diluted with a sufficient amount of water, and then the reaction solution was filtered to obtain a filter cake. The filter cake was slurried with methanol and then dried to obtain Intermediate 1.

[0056] (3) Intermediate 1 (20 mmol) and 4-isothiocyanatophenol (22 mmol) were added to 40 mL of toluene, and the reaction was refluxed in toluene for 2.5 h. After the reaction, the system was cooled to room temperature, and the filter cake was obtained by filtration. The filter cake was washed successively with toluene and n-hexane, and then slurried with ethanol and dried to obtain Intermediate 2. 1 The results of HNMR and MS were consistent with those of Example 1.

[0057] (4) Intermediate 2A (5.5 mmol) and Intermediate 2 (5 mmol) were added to 6 mL of N,N-dimethylformamide (DMF), and potassium carbonate (12 mmol) and copper powder (0.5 mmol) were added as catalysts. The reaction was carried out at 160 °C for 35 min in a microwave reactor (800 W, 2500 HZ). After the reaction was completed, the mixture was cooled to room temperature, and then the reaction was quenched with water. The organic layer was obtained by extraction with ethyl acetate and then dried and column chromatographed to obtain Compound I. 1 The results of HNMR and MS were consistent with those of Example 1.

[0058] Test Example 1

[0059] Cytotoxicity of the compound Ⅰ of the present invention against A549 lung cancer cells

[0060] Adjust the concentration of A549 lung cancer cell suspension in the logarithmic growth phase to 5×10 4 cells / mL, centrifuge and mix to form a single-cell suspension. Add 100 μL to each well of a 96-well culture plate and culture in an incubator at 37 °C and 5% CO2 until the cell confluence reaches 80%, then add the drug for treatment. Add 100 μL of the compound Ⅰ (0.05, 0.10, 0.25, 0.30, 0.50 μM) of Example 1 and DMSO (negative control) to each well and stimulate for 24 h. Add 20 μL of MTT (5 mg / mL) solution to each well and continue to incubate in the cell culture incubator for 4 hours. Measure the absorbance OD value of each well at a wavelength of 490 nm using an enzyme-linked immunosorbent assay reader. This experiment was repeated three times, and the experimental results are as Figure 2 shown.

[0061] It can be seen from Figure 2 that the compound Ⅰ prepared by the present invention can significantly inhibit the proliferation of lung cancer cell A549, and its IC 50 = 0.287 μM, which indicates that the compound Ⅰ has high inhibitory activity against lung cancer cell A549.

[0062] Test Example 2

[0063] Effect of the compound Ⅰ of the present invention on inhibiting EMT of A549 lung cancer cells

[0064] Epithelial-mesenchymal transition (EMT) is a complex cellular process in which cancer cells lose apical-basal polarity and cell-cell adhesion (both characteristics of epithelial cells) and acquire the characteristics of mesenchymal cells. The mesenchymal cells with new motility can migrate from the primary tumor site and spread to secondary sites. The compound Ⅰ inhibits the EMT process of alveolar epithelial cells, thereby inhibiting the migration and invasion of lung cancer cells. EMT is characterized by the loss of epithelial cell markers and the up-regulation of mesenchymal cell marker expression. To investigate the effect of the compound Ⅰ on cell EMT, an EMT model was established by stimulating lung cancer cell A549 with TGF-β. The specific steps for the compound Ⅰ to inhibit the cell EMT are as follows:

[0065] (1) Take non-small cell lung cancer cell A549 in the logarithmic growth phase, digest it with trypsin, centrifuge, resuspend, count, and inoculate it into a 24-well plate at 2.5×10 5 cells / mL, add 500 μL to each well;

[0066] After the cells adhere, starve the cells with EMEM medium containing 0.5% FBS for 12 h;

[0067] (3) Prepare a TGF-β solution with a concentration of 5 ng / mL using EMEM medium containing 0.5% FBS as the TGF-β group; dissolve Compound I of the present invention with DMSO to prepare a Compound I solution with a concentration of 100 mM. Dilute the Compound I solution to the corresponding concentrations (TGF-β + 20 nM Compound I, TGF-β + 50 nM Compound I, TGF-β + 100 nM Compound I) with the medium containing 5 ng / mL TGF-β. Use the medium without TGF-β as the blank control group;

[0068] (4) Finally, aspirate the medium used for starving the cells, and then add the diluted compound into the wells. After culturing for 24 h, wash once with PBS, and add 200 μL of Trizol to lyse and collect the RNA samples;

[0069] (5) Perform RNA transcription using the PrimeScript RT Master Mix kit, and then perform real-time fluorescence quantitative PCR detection using the 2×SYBR Green qPCR Master Mix kit. The inhibitory results of Compound I on cell EMT are as Figures 3 - 6 shown.

[0070] As Figures 3 - 6 can be seen, in A549 cells exposed to TGF-β, the transcriptional expression level of the epithelial marker E-cadherin decreases, and the transcriptional expression levels of the mesenchymal cell markers Vimentin, Fibronectin, and N-cadherin are higher. After treatment with Compound I, the transcriptional expression level of the epithelial marker is significantly increased, and the transcriptional level of the mesenchymal cell marker is inhibited, indicating that Compound I can inhibit the EMT process and has the ability to inhibit the migration and invasion of lung cancer cells.

[0071] Experimental Example 3

[0072] Effect of Compound I of the present invention on the growth of A549 cell xenografts in nude mice

[0073] (1) Prepare a nude mouse xenograft model of non-small cell lung cancer A549 cells

[0074] 40 SPF-grade BALB / c-nu mice, 6 weeks old, with a body weight of 16 - 18 g. Take A549 cells of the lung cancer cell line in the logarithmic growth phase, adjust the concentration of A549 cells to 3×10 7 cells / mL with sterile PBS, inoculate 0.1 mL of A549 cells subcutaneously on the back of BALB / c-nu mice, and wait until the volume of the subcutaneous xenograft reaches about 75 mm 3 or so, then the model is successfully constructed.

[0075] (2) Experimental grouping and administration method

[0076] The successfully modeled mice were randomly divided into the following 3 groups, with 12 mice in each group, and administration started from the 2nd day after successful modeling:

[0077] Example group: Intragastric administration of Compound I prepared in Example 1, with a dose of 25 mg of Compound I / kg / d, once a day, continuously administered for 14 days;

[0078] Gefitinib group: Intragastric administration of gefitinib, with a dose of 25 mg / kg / d, once a day, continuously administered for 14 days;

[0079] Model control group: Intragastric administration of an equal volume of normal saline, once a day, continuously administered for 14 days.

[0080] 48 hours after the last administration, the mice were sacrificed by cervical dislocation, the transplanted tumors were excised, and the tumor weights were weighed. The tumor weight inhibition rate (%) = (1 - mean tumor weight of the test group / mean tumor weight of the model control group) × 100%. The effect of the drug on inhibiting tumor growth in nude mice with human lung cancer A549 cells was reflected by comparing the tumor weights. The data were expressed as mean ± standard deviation and one-way ANOVA was performed using SPSS 15.0 software.

[0081] (3) Results and analysis

[0082] The test results are shown in Table 1.

[0083] Table 1 Tumor weight inhibition rates of Compound I of the present invention and gefitinib on lung cancer

[0084]

[0085] As can be seen from Table 1, Compound I prepared by the present invention has an inhibitory effect on the growth of transplanted tumors of human lung cancer A549 cells in nude mice. Compared with the gefitinib group, the average tumor inhibition rate has increased. This shows that Compound I of the present invention has high efficiency in the treatment of lung cancer, can achieve high-efficiency inhibition of tumor weight with a smaller dose of the drug, and has obtained unexpected technical effects.

[0086] In summary, Compound I prepared by the present invention can significantly inhibit the proliferation of lung cancer cells A549 in mice, and at the same time has an inhibitory effect on the growth of transplanted tumors of human lung cancer A549 cells in nude mice. It has excellent clinical application prospects and provides a new and effective drug administration route for the treatment of lung cancer.

[0087] The above embodiments are only the preferred embodiments of the present invention and cannot be used to limit the scope of protection of the present invention. Any non-substantive changes and substitutions made by those skilled in the art based on the present invention fall within the scope of protection required by the present invention.

Claims

1. Use of compound I in the preparation of a drug for treating lung cancer, characterized in that: The structural formula of the compound I is shown below: 。 2. Use of compound I according to claim 1 in the preparation of a drug for treating lung cancer, characterized in that: The preparation process of the compound I comprises the following steps: (1) Sodium pyruvate is added to an acetic acid solution of 4-bromo-o-phenylenediamine, and the mixture is reacted for 3 to 5 hours and then treated to obtain intermediate 2A; (2) 4-methoxy-α-bromoacetophenone, thiosemicarbazide and inorganic base were mixed evenly, subjected to microwave reaction at 190-205°C for 5-10 min, and then treated to obtain intermediate 1; (3) adding the intermediate 1 and 4-isothiocyanatophenol to a solvent in sequence, reacting under reflux for 2-3 hours, and then treating to obtain the intermediate 2; (4) The intermediate 2, the intermediate 2A, an inorganic base and copper powder are added to a solvent, subjected to microwave reaction at 150-170° C. for 25-35 min, and then treated to obtain compound I.

3. Use of compound I according to claim 2 in the preparation of a drug for treating lung cancer, characterized in that: The dosage ratio of sodium pyruvate, 4-bromo-o-phenylenediamine and acetic acid in step (1) is 1 mmol: (1-1.25) mmol: (2-2.5) mL.

4. Use of compound I according to claim 2 in the preparation of a drug for treating lung cancer, characterized in that: The molar ratio of 4-methoxy-α-bromoacetophenone, thiosemicarbazide and inorganic base in step (2) is 1:(1-1.5):(9-11).

5. Use of compound I according to claim 2 in the preparation of a drug for treating lung cancer, characterized in that: The molar ratio of the intermediate 1 and 4-isothiocyanatophenol in step (3) is 1:(1-1.25).

6. Use of the compound I according to claim 2 in the preparation of a drug for treating lung cancer, characterized in that: The molar ratio of the intermediate 2, intermediate 2A, inorganic base and copper powder in step (4) is 1: (1-1.25): (2-3): (0.1-0.25).

7. Use of the compound I according to claim 2 in the preparation of a drug for treating lung cancer, characterized in that: The inorganic base in steps (2) and (4) is one of cesium carbonate and potassium carbonate.

8. Use of the compound I according to claim 2 in the preparation of a drug for treating lung cancer, characterized in that: The solvent in step (3) is one of toluene and xylene, and the solvent in step (4) is one of N,N-dimethylformamide and N,N-dimethylacetamide.

9. Use of the compound I according to claim 1 in the preparation of a drug for treating lung cancer, characterized in that: The drug also includes pharmaceutically acceptable excipients.

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