Pyruvate preparation for treating non-small cell lung cancer and preparation method thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a pyruvate preparation for treating non-small cell lung cancer and a preparation method of the pyruvate preparation. The pyruvate preparation is prepared from the following components in parts by weight: 0.03 to 0.05 part of sodium pyruvate, 0.01 to 0.03 part of synergist and 100 parts of water for injection. The preparation method of the synergist comprises the following steps: adding sodium pyruvate into a 3, 4-diaminophenol solution for reaction to obtain an intermediate 1; reacting the intermediate 1 with 4-chloro-1, 2-phenylenediamine to obtain an intermediate 2; reacting p-hydroxybenzaldehyde with propynyl chloride to obtain an intermediate 3; reacting the intermediate 3 with 1-azido-4-chlorobenzene to obtain an intermediate 4; and reacting the intermediate 2 with the intermediate 4 to obtain the synergist. Experimental results show that the synergistic agent and the sodium pyruvate are matched to achieve an excellent anti-tumor effect, and particularly, the selectivity of inhibition on non-small cell lung cancer tumors is shown.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a pyruvate preparation for treating non-small cell lung cancer and a preparation method thereof. Background Art

[0002] Lung cancer is one of the most common malignant tumors in China. It is divided into small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) according to pathological types, of which NSCLC accounts for 80% to 85%. Since most lung cancer patients are already in the middle and late stages when they seek medical treatment, they have lost the best opportunity for surgical treatment and can only rely on systemic treatment. Therefore, drug therapy has become the main treatment for advanced NSCLC. Because of this, research on the treatment of lung cancer, especially non-small cell lung cancer, has always been a hot topic and is one of the tumors with the fastest progress.

[0003] In the treatment of non-small cell lung cancer, molecular targeted therapy has become a research hotspot in recent years, such as epidermal growth factor receptor (EGFR) inhibitors, anti-angiogenic drugs, anaplastic lymphoma kinase (ALK) inhibitors, etc. Although targeted therapy has become a relatively new treatment for non-small cell lung cancer, chemotherapy is still the cornerstone of advanced NSCLC treatment. From relevant research reports, it can be seen that for sensitive populations with gene mutations, targeted therapy has better progression-free survival and tolerance than chemotherapy, but it is still difficult for targeted therapy to surpass chemotherapy in terms of overall survival time. The results of a meta-analysis published in the Journal of the American Medical Association in 2004 showed that the effect of two-drug combination therapy based on platinum drugs is better than monotherapy or three-drug combination therapy, thus establishing the status of two-drug combination therapy based on platinum drugs in the treatment of advanced NSCLC. A large number of phase I clinical trials have confirmed that the third-generation chemotherapy drugs combined with cisplatin or carboplatin are effective in the treatment of advanced NSCLC, including paclitaxel, docetaxel, gemcitabine, pemetrexed, vinblastine, vinorelbine, etoposide and albumin paclitaxel, which are widely used in clinical practice. However, the efficacy of chemotherapy for advanced lung cancer is still very limited. Taking the third-generation chemotherapy drugs combined with platinum drugs for the treatment of advanced NSCLC as an example, the objective response rate of treatment is 17% to 22%, the progression-free survival (PFS) of advanced patients is 4 to 6 months, the median survival (OS) is 7.4 to 8.1 months, the 1-year survival rate is 31% to 36%, and the 2-year survival rate is 10% to 13%.

[0004] Based on the above background, researching and developing more new drugs with good tumor-suppressing effects for the treatment of lung cancer, especially non-small cell lung cancer, and expanding treatment options are still urgent issues that need to be addressed in today's medicine. Summary of the invention

[0005] The first object of the present invention is to provide a pyruvate preparation for treating non-small cell lung cancer, which comprises, by weight, 0.03-0.05 parts of sodium pyruvate, 0.01-0.03 parts of a synergist, and 100 parts of water for injection; the structural formula of the synergist is shown in Formula I: Formula I.

[0006] Preferably, the preparation method of the synergist comprises the following steps: (1) adding sodium pyruvate to a 3,4-diaminophenol solution, stirring to react, collecting the precipitate after the reaction is completed, washing and drying to obtain intermediate 1; (2) adding intermediate 1 and 4-chloro-1,2-phenylenediamine to DMF, then adding a catalyst, reacting in an inert gas atmosphere, and after the reaction is completed, post-treating the reaction liquid to obtain intermediate 2; (3) reacting p-hydroxybenzaldehyde with propynyl chloride under catalyst conditions for 12-24 h, and purifying the reaction solution to obtain intermediate 3; (4) adding intermediate 3 and 1-azido-4-chlorobenzene to a mixed solvent of tetrahydrofuran / water, and then adding copper sulfate and sodium ascorbate to react. After the reaction is completed, the reaction solution is purified to obtain intermediate 4; (5) The intermediate 2 and the intermediate 4 are added to ethanol, and then a catalyst is added to carry out a reflux reaction. After the reaction is completed, water is added to the reaction solution, and the precipitated precipitate is collected, washed, and dried. After purification by column chromatography, the synergist is obtained.

[0007] Preferably, in step (1), the molar ratio of 3,4-diaminophenol to sodium pyruvate is 1:(1-1.5), and the stirring reaction time is 8-10 h.

[0008] Preferably, in step (2), the molar ratio of the intermediate 1 to 4-chloro-1,2-phenylenediamine is (1.20-1.51):1; the catalyst is composed of anhydrous potassium carbonate and copper powder; the molar ratio of the intermediate 1, anhydrous potassium carbonate and copper powder is (6-7):(8-9):1.

[0009] Preferably, the reaction temperature in step (2) is 120-140° C., and the reaction time is 24-48 h.

[0010] Preferably, the post-treatment process is: cooling the reaction solution to room temperature, then pouring the reaction solution into water, extracting the product with ether or ethyl acetate, collecting the organic phase, drying it, and then purifying it by column chromatography.

[0011] Preferably, in step (3), the molar ratio of p-hydroxybenzaldehyde, propynyl chloride and catalyst is 1:(1-1.2):(0.5-1.5); the catalyst is potassium carbonate; and the reaction temperature is 20-40°C.

[0012] Preferably, in step (4), the molar ratio of the intermediate 3, 1-azido-4-chlorobenzene, copper sulfate and sodium ascorbate is (5-6): (5-6): 1: (2-2.5); and the reaction time is 12-24 h.

[0013] Preferably, in step (5), the molar ratio of intermediate 2, intermediate 4 and catalyst is 1:(1-1.2):(0.5-1); the catalyst is sodium persulfate.

[0014] The second object of the present invention is to provide a method for preparing the above-mentioned pyruvate preparation for treating non-small cell lung cancer, which is prepared by compounding sodium pyruvate, a synergist and water for injection according to the weight proportions in the above-mentioned first object.

[0015] Compared with the prior art, the beneficial effects of the present invention are mainly: The present invention provides a pyruvate preparation for treating non-small cell lung cancer, comprising sodium pyruvate, a commonly used drug for treating lung diseases, and a special synergist of the present invention, which is prepared by the inventor based on long-term research and molecular structure design and improvement of chemical drugs. Experimental results show that the combination of the synergist and sodium pyruvate has an excellent anti-tumor effect, especially showing selectivity in inhibiting non-small cell lung cancer tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The intermediate 2 of the present invention 1 HNMR spectra; Figure 2 The synergist of the present invention 1 HNMR spectra; Figure 3 is the mass spectrum of the synergist of the present invention; Figure 4 This is a comparison chart of the degradation of EGFR protein expression by the synergist of the present invention and gefitinib. DETAILED DESCRIPTION

[0017] The technical solution of the present invention is further explained below in conjunction with specific embodiments and test examples.

[0018] In the following examples and test examples, the raw materials and preparation methods used are conventional materials and techniques in the art unless otherwise specified.

[0019] Example 1 A pyruvate preparation for treating non-small cell lung cancer, comprising, by weight, 0.04 parts of sodium pyruvate, 0.02 parts of a synergist, and 100 parts of water for injection; The structural formula of the above-mentioned synergist is shown below: ; The preparation method of the above-mentioned synergist comprises the following steps: (1) 3,4-diaminophenol (20 mmol) was dissolved in 50 mL of 1% acetic acid solution, and sodium pyruvate (20 mmol) was dissolved in 30 mL of deionized water. The sodium pyruvate solution was then added dropwise to the 3,4-diaminophenol solution under stirring and reacted at room temperature for 8 h. After the reaction was completed, the precipitate was collected and washed with water three times to remove the unreacted raw material. After drying, the intermediate 1 was obtained.

[0020] The specific reaction formula is as follows: The mass spectrometry analysis of the intermediate 1 was HRMS (ESI + ): [M+H] + Calculated: 177.06, found: 177.06; the above results confirm that the obtained product is the target product.

[0021] (2) Dissolve intermediate 1 (30 mmol) and 4-chloro-1,2-phenylenediamine (25 mmol) in 100 mL dry N,N-dimethylformamide (DMF), add 40 mmol anhydrous potassium carbonate and 5 mmol copper powder as catalysts, and react at 120 °C for 24 h under nitrogen protection. After the reaction is completed, the reaction solution is cooled to room temperature and then poured into a beaker containing 200 mL of water. The product is extracted with ether, the organic layer is collected and dried with anhydrous magnesium sulfate, and after drying, the magnesium sulfate is filtered out and the organic phase is concentrated. The concentrate is purified by column chromatography to obtain intermediate 2.

[0022] The specific reaction formula is as follows: The above intermediate 2 1 The HNMR results are as follows Figure 1 As shown: (C 15 H 14N4O2, 400MHz, DMSO-d6)δ: 10.68(s, 1H), 7.19-7.17(d, 1H), 6.98-6.94(d, 1H), 6.82(s, 1H), 6.65-6.63(d, 1H),6.35(s, 1H), 6.23-6.21(d, 1H), 4.96(s, 4H), 2.19(s, 3H); The above results confirmed that the obtained product was the target product.

[0023] (3) p-Hydroxybenzaldehyde (20 mmol) and propargyl chloride (20 mmol) were added to 100 mL of N,N-dimethylformamide, and then 10 mmol of anhydrous potassium carbonate was added. The mixture was reacted at 20 °C for 12 h. After the reaction was completed, the reaction solution was concentrated, dried and purified by column chromatography to obtain intermediate 3.

[0024] The specific reaction formula is as follows: The mass spectrometry analysis of intermediate 3 was HRMS (ESI + ): [M+H] + Calculated to be 161.05, found to be 161.05; the above results confirmed that the obtained product was the target product.

[0025] (4) Add intermediate 3 (20 mmol) and 1-azido-4-chlorobenzene (20 mmol) to 100 mL of a mixed solvent of tetrahydrofuran: water in a volume ratio of 3:1, then add copper sulfate (4 mmol) and sodium ascorbate (8 mmol), and react at room temperature for 12 h. After the reaction is completed, the reaction solution is concentrated under reduced pressure, then diluted with ethyl acetate, and then washed with water. The organic extracts are combined and dried over anhydrous sodium sulfate, then concentrated under reduced pressure, and purified by column chromatography to obtain intermediate 4.

[0026] The specific reaction formula is as follows: The mass spectrometry analysis of the intermediate 4 was HRMS (ESI + ): [M+H] + Calculated to be 314.06, found to be 314.06; the above results confirmed that the obtained product was the target product.

[0027] (5) Add intermediate 2 (10 mmol) and intermediate 4 (10 mmol) to ethanol, then add sodium persulfate (5 mmol), heat and stir the mixture under reflux, monitor the reaction by TLC, and after the reaction is completed, cool the reaction solution to room temperature and pour it into a beaker filled with ice water. Collect the precipitated crude product, wash it with water, and then dry it in a vacuum dryer, and finally separate it by column chromatography to obtain the synergist.

[0028] The specific reaction formula is as follows: The above synergists 1 HNMR Figure 2 As shown: (C 31 H 22 ClN7O3, 400MHz, DMSO-d6)δ: 12.56(s,1H), 10.66(s, 1H), 8.08(m, 3H), 7.55-7.52(t, 3H), 7.35(d, 2H), 7.18-7.16(m,2H), 7.03-6.94(m, 4H), 6.81(s, 1H), 5.20(s, 2H), 2.07(s, 3H); The mass spectra of the above-mentioned enhancers are shown in Figure 3 As shown, HRMS (ESI + ): [M+H] + Calculated to be 576.15, found to be 576.15; the above results confirmed that the obtained product was the target product.

[0029] This embodiment also provides a method for preparing the above-mentioned pyruvate preparation for treating non-small cell lung cancer, which is prepared by compounding sodium pyruvate, a synergist and water for injection according to the above-mentioned parts by weight.

[0030] Example 2 A pyruvate preparation for treating non-small cell lung cancer, comprising, by weight, 0.03 parts of sodium pyruvate, 0.01 parts of a synergist, and 100 parts of water for injection; The structural formula of the above synergist is as shown in Example 1; The preparation method of the above-mentioned synergist comprises the following steps: (1) 3,4-diaminophenol (20 mmol) was dissolved in 50 mL of 1% acetic acid solution, and sodium pyruvate (24 mmol) was dissolved in 30 mL of deionized water. The sodium pyruvate solution was then added dropwise to the 3,4-diaminophenol solution under stirring and reacted at room temperature for 9 h. After the reaction was completed, the precipitate was collected and washed with water three times to remove the unreacted raw material. After drying, the intermediate 1 was obtained.

[0031] (2) Intermediate 1 (35 mmol) and 4-chloro-1,2-phenylenediamine (25 mmol) were dissolved in 100 mL of dry N,N-dimethylformamide (DMF), and 42 mmol of anhydrous potassium carbonate and 5 mmol of copper powder were added as catalysts. The mixture was reacted at 130 °C for 36 h under nitrogen protection. After the reaction was completed, the reaction solution was cooled to room temperature and then poured into a beaker containing 200 mL of water. The product was extracted with ether, and the organic layer was collected and dried with anhydrous magnesium sulfate. After drying, the magnesium sulfate was removed by filtration and the organic phase was concentrated. The concentrate was purified by column chromatography to obtain intermediate 2.

[0032] (3) p-Hydroxybenzaldehyde (20 mmol) and propargyl chloride (22 mmol) were added to 100 mL of N,N-dimethylformamide, and then 20 mmol of anhydrous potassium carbonate was added. The mixture was reacted at 30 °C for 18 h. After the reaction, the reaction solution was concentrated, dried and purified by column chromatography to obtain intermediate 3.

[0033] (4) Intermediate 3 (24 mmol) and 1-azido-4-chlorobenzene (20 mmol) were added to 100 mL of a mixed solvent of tetrahydrofuran: water in a volume ratio of 3:1, and copper sulfate (4 mmol) and sodium ascorbate (9 mmol) were added. The mixture was reacted at room temperature for 18 h. After the reaction, the reaction solution was concentrated under reduced pressure, diluted with ethyl acetate, and washed with water. The organic extracts were combined and dried over anhydrous sodium sulfate, then concentrated under reduced pressure and purified by column chromatography to obtain intermediate 4.

[0034] (5) Intermediate 2 (10 mmol) and intermediate 4 (12 mmol) were added to ethanol, and sodium persulfate (5 mmol) was then added, and the mixture was heated and stirred under reflux conditions. The reaction was monitored by TLC. After the reaction was completed, the reaction solution was cooled to room temperature and poured into a beaker filled with ice water. The precipitated crude product was collected and washed with water, then dried in a vacuum dryer, and finally separated by column chromatography to obtain the synergist.

[0035] This embodiment also provides a method for preparing the above-mentioned pyruvate preparation for treating non-small cell lung cancer, which is prepared by compounding sodium pyruvate, a synergist and water for injection according to the above-mentioned parts by weight.

[0036] Example 3 A pyruvate preparation for treating non-small cell lung cancer, comprising, by weight, 0.05 parts of sodium pyruvate, 0.03 parts of a synergist, and 100 parts of water for injection; The structural formula of the above synergist is the same as that of Example 1; The preparation method of the above-mentioned synergist comprises the following steps: (1) 3,4-diaminophenol (20 mmol) was dissolved in 50 mL of 1% acetic acid solution, and sodium pyruvate (30 mmol) was dissolved in 40 mL of deionized water. The sodium pyruvate solution was then added dropwise to the 3,4-diaminophenol solution under stirring and reacted at room temperature for 10 h. After the reaction was completed, the precipitate was collected and washed with water three times to remove the unreacted raw material. After drying, the intermediate 1 was obtained.

[0037] (2) Intermediate 1 (35 mmol) and 4-chloro-1,2-phenylenediamine (23.3 mmol) were dissolved in 100 mL of dry N,N-dimethylformamide (DMF), and 45 mmol of anhydrous potassium carbonate and 5 mmol of copper powder were added as catalysts. The reaction was carried out at 140 °C for 48 h under nitrogen protection. After the reaction was completed, the reaction solution was cooled to room temperature and then poured into a beaker containing 200 mL of water. The product was extracted with ethyl acetate, and the organic layer was collected and dried with anhydrous magnesium sulfate. After drying, the magnesium sulfate was removed by filtration and the organic phase was concentrated. The concentrate was purified by column chromatography to obtain intermediate 2.

[0038] (3) p-Hydroxybenzaldehyde (20 mmol) and propargyl chloride (24 mmol) were added to 100 mL of N,N-dimethylformamide, and then 30 mmol of anhydrous potassium carbonate was added. The mixture was reacted at 40 °C for 24 h. After the reaction, the reaction solution was concentrated, dried and purified by column chromatography to obtain intermediate 3.

[0039] (4) Intermediate 3 (24 mmol) and 1-azido-4-chlorobenzene (24 mmol) were added to 100 mL of a mixed solvent of tetrahydrofuran: water in a volume ratio of 3:1, and copper sulfate (4 mmol) and sodium ascorbate (10 mmol) were added, and the mixture was reacted at room temperature for 24 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, diluted with ethyl acetate, and washed with water. The organic extracts were combined and dried over anhydrous sodium sulfate, then concentrated under reduced pressure and purified by column chromatography to obtain intermediate 4.

[0040] (5) Intermediate 2 (10 mmol) and intermediate 4 (12 mmol) were added to ethanol, and sodium persulfate (10 mmol) was then added. The mixture was heated and stirred under reflux conditions, and the reaction was monitored by TLC. After the reaction was completed, the reaction solution was cooled to room temperature and poured into a beaker filled with ice water. The precipitated crude product was collected and washed with water, then dried in a vacuum dryer, and finally separated by column chromatography to obtain the synergist.

[0041] This embodiment also provides a method for preparing the above-mentioned pyruvate preparation for treating non-small cell lung cancer, which is prepared by compounding sodium pyruvate, a synergist and water for injection according to the above-mentioned parts by weight.

[0042] Test Example 1 The inhibitory effect of the synergist of the present invention on the proliferation of lung cancer cells cultured in vitro and its influence on normal human cells Non-small cell lung cancer cells NCI-H1975, HCC827, A549 and normal human liver cells L02 in logarithmic growth phase were inoculated with 3×10 4 Cells were placed on a 96-well culture plate, and after growing for 24 hours, the cells were administered according to the following groups: the Example group or the Gefitinib group was added with different concentrations of the synergist of Example 1 of the present invention or 100 μL of Gefitinib (0.5 μmol / L, 1 μmol / L, 5 μmol / L, 10 μmol / L, 20 μmol / L, 40 μmol / L, 80 μmol / L, 160 μmol / L, 320 μmol / L), and 3 replicate wells were made for each concentration; the control group was added with 100 μL of culture medium. After 72 hours of action, the supernatant was discarded, and 100 μL of serum-free culture medium containing 0.5 mg / mL of MTT was added to each well, and the culture was continued for 4 hours, the supernatant was discarded, and 150 μL of dimethyl sulfoxide was added to each well. The absorbance value (A570) of each well at 570 nm was measured by an ELISA instrument, and the average value of each group was taken to calculate the inhibition rate: inhibition rate = (1-A570 of experimental group / A570 of control group) × 100%, and statistical analysis was performed to calculate IC 50 , the specific results are shown in Table 1.

[0043] Table 1 Effects of potentiators and gefitinib on non-small cell lung cancer cells and normal human cells As can be seen from Table 1, the synergist prepared by the present invention can significantly inhibit the proliferation of non-small cell lung cancer cells NCI-H1975, HCC827, and A549, but has a smaller inhibitory effect on normal human cells L02, showing obvious selectivity. Therefore, it can be used for the treatment of non-small cell lung cancer.

[0044] Test Example 2 Effect of the synergist of the present invention on EGFR protein in non-small cell lung cancer cells NCI-H1975, HCC827 and A549 The immuno Western blot antibody used in this experiment was anti-EGFR (ab52894) (Abcam). The three cells were treated with the enhancer (5 μmol / L) of Example 1 or gefitinib (5 μmol / L) for 48 hours, and 5×10 6Each cell was lysed on ice for 30 minutes with 200 μL RIPA (50 mM Tris pH 8.0, 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1% NP-40) with protease inhibitors, and then centrifuged at 12,000 rpm for 10 minutes. The supernatant was collected and the protein concentration was determined by BCA method. 2×SDS loading buffer was added and boiled at 100°C for 10 minutes. Total protein (100 μg) was loaded and electrophoresed by 15% SDS-PAGE gel according to the molecular weight of the protein, and then transferred to a PVDF membrane (GE healthcare), incubated with the corresponding antibodies, chemiluminescent enzyme substrate HRP Substrate (Millipore), and imaged using the LAS-4000 imaging system (Fuji). The results are shown in Figure 4 .

[0045] from Figure 4 It can be seen that for three different non-small cell lung cancer cells, the enhancer of the present invention can induce EGFR degradation better than gefitinib.

[0046] Test Example 3 Effect of the pyruvate preparation of the present invention on the growth of non-small cell lung cancer A549 cell transplanted tumor in nude mice (1) Preparation of nude mouse non-small cell lung cancer A549 cell transplant tumor model 55 SPF BALB / c-nu mice, 6 weeks old, with a body weight of 16 g-18 g. Take the A549 cells of lung cancer cell line in logarithmic growth phase and adjust the A549 cell concentration to 3×10 7 / mL, 0.1 mL of A549 cells were inoculated subcutaneously on the back of BALB / c-nu mice, and the volume of the subcutaneous transplanted tumor reached 75 mm 3 After about 10 days, the model was successfully manufactured.

[0047] (2) Trial groups and medication methods The mice with successful model establishment were randomly selected and divided into the following 4 groups, with 12 mice in each group, and the drug was administered starting from the second day after successful model establishment.

[0048] Group 1-3: The pyruvate preparation prepared in Example 1-3 was administered by atomization at a dose of 0.6 mg sodium pyruvate / kg / d, once a day, for 14 consecutive days; Gefitinib group: gefitinib was administered orally at a dose of 25 mg / kg / d, once a day, for 14 consecutive days; Model control group: gavage with an equal amount of normal saline once a day for 14 consecutive days.

[0049] Sodium pyruvate group: Sodium pyruvate solution was administered by nebulization, with a dose of 0.6 mg sodium pyruvate / kg / d, once a day, for 14 consecutive days; 48 hours after the last administration, mice were killed by dislocation, transplanted tumors were removed, tumor weights of each mouse were weighed, and the average tumor weight of each group was calculated. Tumor weight inhibition rate (%) = (1-mean tumor weight of the experimental group / mean tumor weight of the model control group) × 100%. The effect of the drug on the inhibition of tumor growth in nude mice with human lung cancer A549 cells was reflected by comparison of tumor weight. Data were expressed as mean ± standard deviation (x ± s, n = 12), and variance analysis was performed using SPSS15.0 software.

[0050] (3) Results and analysis The test results are shown in Table 2: Table 2 The tumor weight inhibition rate of pyruvate preparation and gefitinib on non-small cell lung cancer xenografts in nude mice From the results in Table 2, it can be seen that the pyruvate preparation prepared by the present invention has an inhibitory effect on the growth of human lung cancer A549 cell nude mouse transplanted tumors. Compared with the gefitinib group and the sodium pyruvate group, the average tumor inhibition rate is improved. This shows that the synergist prepared by the present invention has a very good therapeutic effect and low toxic side effects in the treatment of non-small cell lung cancer after being combined with sodium pyruvate, and an unexpected technical effect is obtained.

[0051] In summary, the enhancer prepared by the present invention can significantly inhibit the proliferation of non-small cell lung cancer cells NCI-H1975, HCC827, and A549, and the pyruvate preparation added with the enhancer has an inhibitory effect on the growth of human lung cancer A549 cell nude mouse transplanted tumors, has excellent clinical application prospects, and provides a new and effective medication route for the treatment of non-small cell lung cancer.

[0052] The above are only preferred embodiments of the present invention, and are not limited to the above examples. For those skilled in the art, various changes and variations are possible under the principle of the present invention. Any modifications and improvements made should be considered within the scope of protection of the present invention.

Claims

1. A pyruvate preparation for treating non-small cell lung cancer, characterized in that: In parts by weight, it comprises 0.03-0.05 parts of sodium pyruvate, 0.01-0.03 parts of a synergist, and 100 parts of water for injection; the structural formula of the synergist is shown in Formula I: Formula I.

2. The pyruvate preparation for treating non-small cell lung cancer according to claim 1, characterized in that: The preparation method of the synergist comprises the following steps: (1) adding sodium pyruvate to a 3,4-diaminophenol solution, stirring to react, collecting the precipitate after the reaction is complete, washing and drying to obtain intermediate 1; (2) adding intermediate 1 and 4-chloro-1,2-phenylenediamine to DMF, then adding a catalyst, reacting in an inert gas atmosphere, and after the reaction is completed, post-treating the reaction solution to obtain intermediate 2; (3) reacting p-hydroxybenzaldehyde with propynyl chloride under catalyst conditions for 12-24 h, and purifying the reaction solution to obtain intermediate 3; (4) adding intermediate 3 and 1-azido-4-chlorobenzene to a mixed solvent of tetrahydrofuran / water, and then adding copper sulfate and sodium ascorbate to react. After the reaction is completed, the reaction solution is purified to obtain intermediate 4; (5) The intermediate 2 and the intermediate 4 are added to ethanol, and then a catalyst is added to carry out a reflux reaction. After the reaction is completed, water is added to the reaction solution, and the precipitated precipitate is collected, washed, and dried. After purification by column chromatography, the synergist is obtained.

3. The pyruvate preparation for treating non-small cell lung cancer according to claim 2, characterized in that: In step (1), the molar ratio of 3,4-diaminophenol to sodium pyruvate is 1:(1-1.5), and the stirring reaction time is 8-10 hours.

4. The pyruvate preparation for treating non-small cell lung cancer according to claim 2, characterized in that: The molar ratio of the intermediate 1 and 4-chloro-1,2-phenylenediamine in step (2) is (1.20-1.51):1; the catalyst is composed of anhydrous potassium carbonate and copper powder; the molar ratio of the intermediate 1, anhydrous potassium carbonate and copper powder is (6-7):(8-9):

1.

5. The pyruvate preparation for treating non-small cell lung cancer according to claim 2, characterized in that: The reaction temperature in step (2) is 120-140° C., and the reaction time is 24-48 h.

6. The pyruvate preparation for treating non-small cell lung cancer according to claim 2, characterized in that: The post-treatment process is as follows: cooling the reaction solution to room temperature, then pouring the reaction solution into water, extracting the product with ether or ethyl acetate, collecting the organic phase, drying it, and then purifying it by column chromatography.

7. The pyruvate preparation for treating non-small cell lung cancer according to claim 2, characterized in that: In step (3), the molar ratio of p-hydroxybenzaldehyde, propynyl chloride and catalyst is 1:(1-1.2):(0.5-1.5); the catalyst is potassium carbonate; and the reaction temperature is 20-40°C.

8. The pyruvate preparation for treating non-small cell lung cancer according to claim 2, characterized in that: In step (4), the molar ratio of the intermediate 3, 1-azido-4-chlorobenzene, copper sulfate and sodium ascorbate is (5-6): (5-6): 1: (2-2.5); the reaction time is 12-24 h.

9. The pyruvate preparation for treating non-small cell lung cancer according to claim 2, characterized in that: In step (5), the molar ratio of intermediate 2, intermediate 4 and catalyst is 1:(1-1.2):(0.5-1); the catalyst is sodium persulfate.

10. The method for preparing a pyruvate preparation for treating non-small cell lung cancer according to any one of claims 1 to 9, characterized in that: Sodium pyruvate, synergist and water for injection are compounded according to the above parts by weight to obtain the product.

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