A pyruvate preparation for treating non-small cell lung cancer and its preparation method

Abstract

The present invention belongs to the technical field of biological medicine, and particularly relates to a pyruvate preparation for treating non-small cell lung cancer and a preparation method thereof. The pyruvate preparation 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 preparation method of the synergist comprises: adding sodium pyruvate to a 3,4-diaminophenol solution for reaction to obtain intermediate 1; reacting intermediate 1 with 4-chloro-1,2-phenylenediamine to obtain intermediate 2; reacting p-hydroxybenzaldehyde with propargyl chloride to obtain intermediate 3; reacting intermediate 3 with 1-azido-4-chlorobenzene to obtain intermediate 4; and then reacting intermediate 2 and intermediate 4 to obtain the synergist. Experimental results show that the combination of the synergist and sodium pyruvate has excellent anti-tumor effects, especially showing selectivity for inhibiting non-small cell lung cancer tumors.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly 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. According to the pathological type, it is divided into small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC), among which NSCLC accounts for 80% - 85%. Since most lung cancer patients are in the middle and late stages when they seek medical treatment, they have missed the best opportunity for surgical treatment and can only rely on systemic treatment. Therefore, drug treatment has become the main treatment method for advanced NSCLC. For this reason, research on the treatment methods of lung cancer, especially the treatment of non-small cell lung cancer, has always been a hot topic and one of the tumors with the most rapid progress.

[0003] In the treatment methods of non-small cell lung cancer, in recent years, molecular targeted therapy has become a research hotspot, such as epidermal growth factor receptor (EGFR) inhibitors, anti-angiogenic drugs, anaplastic lymphoma kinase (ALK) inhibitors, etc. Although targeted therapy has become a relatively novel therapy for the treatment of non-small cell lung cancer, chemotherapy is still the cornerstone of the treatment of advanced NSCLC. It can be seen from relevant research reports that for sensitive populations with gene mutations, the progression-free survival and tolerance of targeted therapy are better than those of chemotherapy, but in terms of overall survival time, targeted therapy is still difficult to surpass chemotherapy. The results of a Meta-analysis 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 that of single-drug therapy or three-drug combination therapy, thus establishing the status of the 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 combination of the third-generation chemotherapy drugs and cisplatin or carboplatin is effective in the treatment of advanced NSCLC. Among them, chemotherapy drugs such as paclitaxel, docetaxel, gemcitabine, pemetrexed, vinblastine, vinorelbine, etoposide, and albumin-bound paclitaxel are widely used in clinical practice. However, the efficacy of chemotherapy for advanced lung cancer is still very limited. Taking the combination of the third-generation chemotherapy drugs and platinum drugs for the treatment of advanced NSCLC as an example, its objective remission rate for treatment is 17% - 22%, the progression-free survival (PFS) of advanced patients is 4 - 6 months, the median survival time (OS) is 7.4 - 8.1 months, the 1-year survival rate is 31% - 36%, and the 2-year survival rate is 10% - 13%.

[0004] Based on the above background, researching and developing more new drugs with good anti-tumor effects for treating lung cancer, especially non-small cell lung cancer, and expanding treatment approaches are still urgent problems to be solved in current medicine. Summary of the Invention

[0005] The first object of the present invention is to provide a pyruvate preparation for the treatment of 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 as shown in Formula I:

[0006] Formula I.

[0007] Preferably, the preparation method of the synergist comprises the following steps:

[0008]

[0009] (1) Sodium pyruvate is added to a 3,4-diaminophenol solution, and the mixture is stirred and reacted. After the reaction is completed, the precipitate is collected, washed, and dried to obtain Intermediate 1;

[0010] (2) Intermediate 1 and 4-chloro-1,2-phenylenediamine are added to DMF, and then a catalyst is added. The reaction is carried out in an inert gas atmosphere. After the reaction is completed, the reaction solution is post-treated to obtain Intermediate 2;

[0011] (3) p-Hydroxybenzaldehyde and propargyl chloride are reacted for 12 - 24 h under the condition of a catalyst. After the reaction solution is purified, Intermediate 3 is obtained;

[0012] (4) Intermediate 3 and 1-azido-4-chlorobenzene are added to a mixed solvent of tetrahydrofuran / water, and then copper sulfate and sodium ascorbate are added for reaction. After the reaction is completed, the reaction solution is purified to obtain Intermediate 4;

[0013] (5) Intermediate 2 and Intermediate 4 are added to ethanol, and then a catalyst is added for reflux reaction. After the reaction is completed, water is added to the reaction solution, and the precipitated solid is collected, washed, and dried. After purification by column chromatography, the synergist is obtained.

[0014] 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.

[0015] Preferably, in step (2), the molar ratio of 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 Intermediate 1, anhydrous potassium carbonate, and copper powder is (6 - 7):(8 - 9):1.

[0016] Preferably, in step (2), the reaction temperature is 120 - 140 °C, and the reaction time is 24 - 48 h.

[0017] Preferably, the post-treatment process is as follows: the reaction solution is cooled to room temperature, then poured into water, and the product is extracted with ether or ethyl acetate. The organic phase is collected, dried, and then purified by column chromatography.

[0018] Preferably, in step (3), the molar ratio of p-hydroxybenzaldehyde, propargyl chloride to the catalyst is 1: (1 - 1.2): (0.5 - 1.5); the catalyst is potassium carbonate; the reaction temperature is 20 - 40 °C.

[0019] Preferably, in step (4), the molar ratio of 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.

[0020] Preferably, in step (5), the molar ratio of intermediate 2, intermediate 4 to the catalyst is 1: (1 - 1.2): (0.5 - 1); the catalyst is sodium persulfate.

[0021] The second object of the present invention is to provide a preparation method of the pyruvate preparation for treating non-small cell lung cancer as described above. Sodium pyruvate, a synergist and water for injection are compounded according to the weight parts in the first object above, and then the product is obtained.

[0022] Compared with the prior art, the beneficial effects of the present invention mainly lie in:

[0023] The present invention provides a pyruvate preparation for treating non-small cell lung cancer, which includes sodium pyruvate, a common drug for treating lung diseases, and a synergist specially prepared by the present invention. The synergist is prepared by the inventor based on long-term research and the molecular structure design and improvement of chemical drugs. Experimental results show that the combination of the synergist and sodium pyruvate has excellent anti-tumor effects, especially showing selectivity in inhibiting non-small cell lung cancer tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is the 1 HNMR spectrum of intermediate 2 of the present invention;

[0025] Figure 2 It is the 1 HNMR spectrum of the synergist of the present invention;

[0026] Figure 3 It is the mass spectrum of the synergist of the present invention;

[0027] Figure 4 It is the comparison chart of the degradation of EGFR protein expression by the synergist and gefitinib of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0028] The technical solution of the present invention will be further explained and illustrated below in conjunction with specific embodiments and test examples.

[0029] In the following examples and test examples, unless otherwise specified, the raw materials and preparation methods used are conventional materials and techniques in the art.

[0030] Example 1

[0031] A pyruvate preparation for treating non-small cell lung cancer, by weight, comprises 0.04 parts of sodium pyruvate, 0.02 parts of synergist, and 100 parts of water for injection;

[0032] The structural formula of the above synergist is as follows:

[0033] ;

[0034] The preparation method of the above synergist includes the following steps:

[0035] (1) Dissolve 3,4-diaminophenol (20 mmol) in 50 mL of 1% acetic acid solution, dissolve sodium pyruvate (20 mmol) in 30 mL of deionized water, and then drop the sodium pyruvate solution into the 3,4-diaminophenol solution under stirring conditions, and react at room temperature for 8 h; after the reaction is completed, collect the precipitate and wash it 3 times with water to remove unreacted raw materials; after drying, intermediate 1 is obtained.

[0036] The specific reaction formula is as follows:

[0037]

[0038] The mass spectrometry analysis HRMS (ESI + ) of the above intermediate 1: [M+H] + Calculated to be 177.06, found to be 177.06; the above results confirm that the obtained product is the target product.

[0039] (2) Dissolve intermediate 1 (30 mmol) and 4-chloro-1,2-phenylenediamine (25 mmol) in 100 mL of dry N,N-dimethylformamide (DMF), add 40 mmol of anhydrous potassium carbonate and 5 mmol of copper powder as catalysts, and react at 120 °C for 24 h under nitrogen protection. After the reaction is completed, cool the reaction solution to room temperature, and then pour it into a beaker containing 200 mL of water. Extract the product with ether, collect the organic layer and dry it with anhydrous magnesium sulfate. After drying, filter to remove magnesium sulfate and concentrate the organic phase. The concentrated solution is purified by column chromatography to obtain intermediate 2.

[0040] The specific reaction formula is as follows:

[0041]

[0042] For the above intermediate 2 1 The HNMR results are as Figure 1 follows: (C 15 H 14 N4O2, 400 MHz, 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 confirm that the obtained product is the target product.

[0043] (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 reaction was carried out 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.

[0044] The specific reaction formula is as follows:

[0045]

[0046] The HRMS (ESI + ) of the above intermediate 3: [M + H] + Calculated: 161.05, Found: 161.05; The above results confirm that the obtained product is the target product.

[0047] (4) Intermediate 3 (20 mmol) and 1-azido-4-chlorobenzene (20 mmol) were added to 100 mL of a mixed solvent of tetrahydrofuran:water with a volume ratio of 3:1, and then copper sulfate (4 mmol) and sodium ascorbate (8 mmol) were added. The reaction was carried out at room temperature for 12 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, then diluted with ethyl acetate, and washed with water. The combined organic extracts were dried with anhydrous sodium sulfate, then concentrated under reduced pressure, and purified by column chromatography to obtain intermediate 4.

[0048] The specific reaction formula is as follows:

[0049]

[0050] The HRMS (ESI + ) of the above intermediate 4: [M + H] + Calculated: 314.06, Found: 314.06; The above results confirm that the obtained product is the target product.

[0051] (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 conditions. Monitor the reaction by TLC. After the reaction is completed, cool the reaction solution to room temperature and pour it into a beaker containing ice water. Collect the precipitated crude product, wash it with water, then dry it in a vacuum desiccator, and finally separate it by column chromatography to obtain the synergist.

[0052] The specific reaction formula is as follows:

[0053]

[0054] The above-mentioned synergist's 1 HNMR is as Figure 2 shown: (C 31 H 22 ClN7O3, 400 MHz, 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 spectrum of the above-mentioned synergist is as Figure 3 shown, HRMS (ESI + ) : [M + H] + Calculated: 576.15, Found: 576.15; The above results confirm that the obtained product is the target product.

[0055] This example also provides a preparation method of the above-mentioned pyruvate preparation for treating non-small cell lung cancer. Compound sodium pyruvate, the synergist and water for injection according to the above weight parts, and then the product can be obtained.

[0056] Example 2

[0057] A pyruvate preparation for treating non-small cell lung cancer, by weight, includes 0.03 parts of sodium pyruvate, 0.01 part of the synergist, and 100 parts of water for injection;

[0058] The structural formula of the above-mentioned synergist is as in Example 1;

[0059] The preparation method of the above-mentioned synergist includes the following steps:

[0060] (1) Dissolve 3,4-diaminophenol (20 mmol) in 50 mL of 1% acetic acid solution, and dissolve sodium pyruvate (24 mmol) in 30 mL of deionized water. Then, under stirring conditions, drop the sodium pyruvate solution into the 3,4-diaminophenol solution and react at room temperature for 9 h. After the reaction is completed, collect the precipitate and wash it with water three times to remove the unreacted raw materials. After drying, intermediate 1 is obtained.

[0061] (2) Dissolve intermediate 1 (35 mmol) and 4-chloro-1,2-phenylenediamine (25 mmol) in 100 mL of dry N,N-dimethylformamide (DMF), add 42 mmol of anhydrous potassium carbonate and 5 mmol of copper powder as catalysts, and react at 130 °C for 36 h under nitrogen protection. After the reaction is completed, cool the reaction solution to room temperature, and then pour it into a beaker containing 200 mL of water. Extract the product with ether, collect the organic layer and dry it with anhydrous magnesium sulfate. After drying, filter to remove magnesium sulfate and concentrate the organic phase. The concentrated solution is purified by column chromatography to obtain intermediate 2.

[0062] (3) Add p-hydroxybenzaldehyde (20 mmol) and propargyl chloride (22 mmol) to 100 mL of N,N-dimethylformamide, and then add 20 mmol of anhydrous potassium carbonate. React at 30 °C for 18 h. After the reaction is completed, the reaction solution is concentrated, dried and purified by column chromatography to obtain intermediate 3.

[0063] (4) Add intermediate 3 (24 mmol) and 1-azido-4-chlorobenzene (20 mmol) to 100 mL of a mixed solvent of tetrahydrofuran:water with a volume ratio of 3:1, then add copper sulfate (4 mmol) and sodium ascorbate (9 mmol), and react at room temperature for 18 h. After the reaction is completed, concentrate the reaction solution under reduced pressure, then dilute it with ethyl acetate and wash it with water. Combine the organic extracts and dry them with anhydrous sodium sulfate, then concentrate under reduced pressure and purify by column chromatography to obtain intermediate 4.

[0064] (5) Add intermediate 2 (10 mmol) and intermediate 4 (12 mmol) to ethanol, then add sodium persulfate (5 mmol), and heat and stir the mixture under reflux conditions. Monitor the reaction by TLC. After the reaction is completed, cool the reaction solution to room temperature and pour it into a beaker containing ice water. Collect the precipitated crude product and wash it with water, then dry it in a vacuum dryer, and finally separate it by column chromatography to obtain the synergist.

[0065] This example also provides a preparation method of the above-mentioned pyruvate preparation for treating non-small cell lung cancer. Compound sodium pyruvate, the synergist and water for injection according to the above weight parts to obtain it.

[0066] Example 3

[0067] A pyruvate preparation for treating non-small cell lung cancer, by weight, comprises 0.05 parts of sodium pyruvate, 0.03 parts of synergist, and 100 parts of water for injection;

[0068] The structural formula of the above synergist is the same as that in Example 1;

[0069] The preparation method of the above synergist comprises the following steps:

[0070] (1) Dissolve 3,4-diaminophenol (20 mmol) in 50 mL of 1% acetic acid solution, dissolve sodium pyruvate (30 mmol) in 40 mL of deionized water, and then drop the sodium pyruvate solution into the 3,4-diaminophenol solution under stirring conditions, and react at room temperature for 10 h; after the reaction is completed, collect the precipitate and wash it with water 3 times to remove unreacted raw materials; after drying, intermediate 1 is obtained.

[0071] (2) Dissolve intermediate 1 (35 mmol) and 4-chloro-1,2-phenylenediamine (23.3 mmol) in 100 mL of dry N,N-dimethylformamide (DMF), add 45 mmol of anhydrous potassium carbonate and 5 mmol of copper powder as catalysts, and react at 140 °C for 48 h under nitrogen protection. After the reaction is completed, cool the reaction solution to room temperature, and then pour it into a beaker containing 200 mL of water. Extract the product with ethyl acetate, collect the organic layer and dry it with anhydrous magnesium sulfate, filter off the magnesium sulfate after drying and concentrate the organic phase, and purify the concentrated solution by column chromatography to obtain intermediate 2.

[0072] (3) Add p-hydroxybenzaldehyde (20 mmol) and propargyl chloride (24 mmol) to 100 mL of N,N-dimethylformamide, then add 30 mmol of anhydrous potassium carbonate, and react at 40 °C for 24 h. After the reaction is completed, concentrate, dry and purify the reaction solution by column chromatography to obtain intermediate 3.

[0073] (4) Add intermediate 3 (24 mmol) and 1-azido-4-chlorobenzene (24 mmol) to 100 mL of a mixed solvent with a volume ratio of tetrahydrofuran to water of 3:1, then add copper sulfate (4 mmol) and sodium ascorbate (10 mmol), and react at room temperature for 24 h. After the reaction is completed, concentrate the reaction solution under reduced pressure, then dilute it with ethyl acetate and wash it with water. Combine the organic extracts and dry them with anhydrous sodium sulfate, then concentrate under reduced pressure and purify by column chromatography to obtain intermediate 4.

[0074] (5) Add intermediate 2 (10 mmol) and intermediate 4 (12 mmol) to ethanol, then add sodium persulfate (10 mmol). Heat and stir the mixture under reflux conditions. Monitor the reaction by TLC. After the reaction is completed, cool the reaction solution to room temperature and pour it into a beaker containing ice water. Collect the precipitated crude product, wash it with water, then dry it in a vacuum dryer, and finally separate it by column chromatography to obtain the synergist.

[0075] This example also provides a preparation method of the above-mentioned pyruvate preparation for treating non-small cell lung cancer. Compound sodium pyruvate, the synergist and water for injection according to the above weight parts to obtain it.

[0076] Test Example 1

[0077] Inhibitory effect of the synergist of the present invention on the proliferation of lung cancer cells cultured in vitro and its effect on human normal cells

[0078] Take non-small cell lung cancer cells NCI-H1975, HCC827, A549 and human normal liver cells L02 in the logarithmic growth phase, and inoculate 3×10 4 cells on a 96-well culture plate. After growing for 24 h, administer drugs according to the following groups: the example group or the gefitinib group is added with different concentrations of the synergist or gefitinib of Example 1 of the present invention, 100 μL (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 each concentration is made into 3 duplicate wells; the control group is added with 100 μL of culture medium. After acting for 72 h, discard the supernatant, add 100 μL of serum-free culture medium containing 0.5 mg / mL MTT to each well, and continue to culture for 4 h. Discard the supernatant, and add 150 μL of dimethyl sulfoxide to each well. Measure the absorbance value (A570) of each well at 570 nm with an enzyme-linked immunosorbent assay instrument. Take the average value of each group and calculate the inhibition rate: Inhibition rate = (1 - A570 of the experimental group / A570 of the control group) × 100%, and perform statistical analysis and calculate IC 50 , and the specific results are shown in Table 1.

[0079] Table 1 Effects of the synergist and gefitinib on non-small cell lung cancer cells and human normal cells

[0080]

[0081] 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, A549, while having a relatively small inhibitory effect on human normal cells L02, showing obvious selectivity. Therefore, it can be used for the treatment of non-small cell lung cancer.

[0082] Experimental Example 2

[0083] Effect of the synergist of the present invention on EGFR protein in non-small cell lung cancer cells NCI-H1975, HCC827, and A549

[0084] The immunoblot antibody used in this experimental example was anti-EGFR (ab52894) (Abcam). After treating the three types of cells with the synergist (5 μmol / L) or gefitinib (5 μmol / L) of Example 1 for 48 hours, 5×10 6 cells were collected and lysed on ice for 30 minutes with 200 μL of RIPA (50 mM Tris pH 8.0, 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1% NP-40) supplemented with protease inhibitors. After centrifugation at 12,000 rpm for 10 minutes, the supernatant was collected. The protein concentration was determined by the BCA method, and 2×SDS loading buffer was added and boiled at 100°C for 10 minutes. Total protein (100 μg) was loaded, electrophoresed on a 15% SDS-PAGE gel according to the molecular weight of the protein, and then transferred to a PVDF membrane (GE healthcare). The corresponding antibodies were used for incubation respectively, and the chemiluminescent enzyme substrate HRP Substrate (Millipore) was used. Imaging was performed using a LAS-4000 imaging system (Fuji). The results are shown in Figure 4 .

[0085] It can be seen from Figure 4 that for the three different non-small cell lung cancer cells, the synergist of the present invention can induce EGFR degradation better than gefitinib.

[0086] Experimental Example 3

[0087] Effect of the pyruvate preparation of the present invention on the growth of nude mouse xenografts of non-small cell lung cancer A549 cells

[0088] (1) Preparation of a nude mouse xenograft model of non-small cell lung cancer A549 cells

[0089] Fifty-five SPF-grade BALB / c-nu mice, 6 weeks old, with a body weight of 16 g - 18 g. Logarithmic growth phase lung cancer cell line A549 cells were taken, and the concentration of A549 cells was adjusted to 3×10 7 / mL with sterile PBS. 0.1 mL of A549 cells was subcutaneously inoculated into the back of BALB / c-nu mice. When the volume of the subcutaneous xenograft reached about 75 mm 3 (about 10 days), the model was successfully established.

[0090] (2) Experimental grouping and administration method

[0091] Successfully modeled mice were randomly selected and divided into the following 4 groups, with 12 mice in each group, and administration started on the second day after successful modeling.

[0092] Groups of Examples 1 - 3: The pyruvate preparations prepared in Examples 1 - 3 were administered by atomization, with a dose of 0.6 mg sodium pyruvate / kg / d, once a day, for 14 consecutive days;

[0093] Gefitinib group: Gefitinib was administered by gavage, with a dose of 25 mg / kg / d, once a day, for 14 consecutive days;

[0094] Model control group: An equal volume of normal saline was administered by gavage, once a day, for 14 consecutive days.

[0095] Sodium pyruvate group: A sodium pyruvate solution was administered by atomization, with a dose of 0.6 mg sodium pyruvate / kg / d, once a day, for 14 consecutive days;

[0096] 48 h after the last administration, the mice were sacrificed by cervical dislocation, the transplanted tumors were excised, the tumor weights of each mouse were weighed, and the average tumor weight of each group was calculated. The 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 inhibiting the growth of transplanted tumors of human lung cancer A549 cells in nude mice was reflected by comparing the tumor weights. The data were expressed as mean ± standard deviation (x ± s, n = 12), and one - way ANOVA was performed using SPSS15.0 software.

[0097] (3) Results and analysis

[0098] The test results are shown in Table 2:

[0099] Table 2 Tumor weight inhibition rates of pyruvate preparations and gefitinib on transplanted tumors of non - small cell lung cancer in nude mice

[0100]

[0101] 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 transplanted tumors of human lung cancer A549 cells in nude mice. Compared with the gefitinib group and the sodium pyruvate group, the average tumor inhibition rate has increased. This shows that the synergist prepared by the present invention combined with sodium pyruvate has very good curative effects and low toxic and side effects in the treatment of non - small cell lung cancer, and unexpected technical effects are obtained.

[0102] In summary, the synergist prepared by the present invention can significantly inhibit the proliferation of non-small cell lung cancer cells NCI-H1975, HCC827, and A549. The pyruvate preparation added with the synergist has an inhibitory effect on the growth of xenografted tumors of human lung cancer A549 cells in nude mice, and has excellent clinical application prospects, providing a new and effective drug administration route for the treatment of non-small cell lung cancer.

[0103] The above are only the preferred embodiments of the present invention and are not limited to the above examples. For those skilled in the art, various changes and modifications can be made under the principle of the present invention. Any modifications, improvements, etc. should be regarded as within the protection scope of the present invention.

Claims

1. A pyruvate preparation for treating non-small cell lung cancer, characterized in that: The pyruvate preparation is composed of the following components in parts 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.

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 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.

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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