Application of pyruvate in preparation of medicine for treating lung biofilm infection

Abstract

The invention belongs to the field of biological medicine, and particularly relates to application of pyruvate in preparation of a medicine for treating lung biofilm infection. According to the application of the pyruvate in preparation of the medicine for treating lung biofilm infection, the pyruvate is a compound I, and the structural formula of the pyruvate is shown in the specification. The pyruvate disclosed by the invention is prepared from sodium pyruvate through chemical modification, has anti-infection activity and biocompatibility, and can be used for remarkably reducing the expression level of inflammatory factors and improving the treatment effect while reducing the dosage of medicines. The pyruvate disclosed by the invention has a wide application prospect in preparation of a medicine for treating lung biofilm infection.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine, and specifically relates to the application of pyruvate in the preparation of a drug for treating lung biofilm infection. Background Art

[0002] Biofilm infection can cause pulmonary inflammation, which can cause breathing difficulties, lung tissue damage and airway obstruction, and even pneumonia. Especially when Pseudomonas aeruginosa invades the human lungs, it can cause severe lung inflammation, leading to symptoms such as high fever, cough, and sputum, and the sputum is often yellow-green, sticky and smelly. In addition, Pseudomonas aeruginosa can also produce a variety of virulence factors, such as endotoxins and exotoxins, which further aggravate lung tissue damage and may even cause serious complications such as lung abscesses and empyema, posing a serious threat to the patient's life and health. Therefore, lung infection caused by Pseudomonas aeruginosa is a serious disease that requires timely diagnosis and active treatment. Antibiotics are generally used to treat biofilm infections, but due to the protective effect of biofilms, antibiotics are difficult to penetrate into the biofilm to kill bacteria, resulting in poor treatment effects; and it is easy to cause drug resistance, further exacerbating the difficulty of treatment.

[0003] As an important organic compound, sodium pyruvate has good antioxidant and anti-inflammatory effects in vivo, which can effectively reduce inflammatory response and protect lung tissue. However, sodium pyruvate also has certain defects, such as: sodium pyruvate has a relatively single functionality, poor therapeutic effect and a narrow application range.

[0004] Therefore, developing pyruvate with new efficacy, wider application range and indications is of great significance to effectively improve the therapeutic effect of the drug. Summary of the invention

[0005] One of the purposes of the present invention is to provide the use of pyruvate in the preparation of a drug for treating lung biofilm infection, wherein the pyruvate is compound I, and its structural formula is as follows: Compound I.

[0006] Furthermore, the preparation steps of the pyruvate are as follows: (1) Weighing 4-chlorophenylhydrazine and 3-methyl-2-butanone into ethanol, and then adding sulfuric acid to obtain a mixed solution, subjecting the mixed solution to reflux reaction. After the reaction is completed, adjusting the pH of the reaction solution to 8.5-9.0, and performing post-treatment to obtain intermediate 1; (2) Adding the DMF solution of intermediate 1 to the DMF solution of POCl3, heating for reaction, and after completion of the reaction, post-treatment to obtain intermediate 2; (3) adding intermediate 2, 3-cyanophenylhydrazine and glacial acetic acid to ethanol, and then reacting under reflux. After the reaction is completed, the reaction solution is cooled to room temperature, filtered, washed, dried and purified to obtain intermediate 3; (4) Dissolving 3,4-dihydroxybenzoic acid and sodium pyruvate in ethanol, then adding KOH ethanol solution thereto, stirring for reaction, and after completion of the reaction, post-treatment to obtain intermediate 4; (5) adding intermediate 4 and 4-formylphenylboronic acid to an aqueous solution of ethanol, adjusting the pH of the mixed solution to 8.2, stirring the mixture for reaction, and after the reaction is completed, purifying the reaction solution to obtain intermediate 5; (6) Add intermediate 3 and intermediate 5 to an aqueous solution of ethanol and stir to react. After the reaction is completed, concentrate, dry and purify the reaction solution to obtain the final product, i.e., compound I.

[0007] Furthermore, in step (1), the molar ratio of 4-chlorophenylhydrazine, 3-methyl-2-butanone and sulfuric acid is (1-1.2):1:(1.2-1.8), and the reflux reaction time is 1-2 h.

[0008] Furthermore, in the DMF solution of the intermediate 1 in step (2), the amount ratio of the intermediate 1 to DMF is 10.32 mmol: 5 mL; in the DMF solution of POCl3, the amount ratio of POCl3 to DMF is 36.12 mmol: 5.5 mL; the molar ratio of the intermediate 1 to POCl3 is 1: (3-4); the temperature of the heating reaction is 70-80°C, and the heating reaction time is 3-5 h.

[0009] Furthermore, in step (3), the molar ratio of the intermediate 2, 3-cyanophenylhydrazine and glacial acetic acid is 1:(1.1-1.3):(2-3), and the reflux time is 18-20 h.

[0010] Furthermore, in step (4), the molar ratio of 3,4-dihydroxybenzoic acid, sodium pyruvate and KOH is 1:(1-1.5):(1.5-2), and the reaction time is 5-7 h.

[0011] Furthermore, in step (5), the molar ratio of the intermediate 4 to 4-formylphenylboronic acid is 1:(1-1.5), and the reaction time is 7-9 h.

[0012] Furthermore, in step (6), the molar ratio of intermediate 3 to intermediate 5 is 1:(1-1.5); and the reaction time is 12-14 h.

[0013] Furthermore, the pulmonary biofilm infection is a pulmonary biofilm infection caused by Pseudomonas aeruginosa.

[0014] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides the use of pyruvate in the preparation of a drug for treating lung biofilm infection. The present invention prepares a pyruvate compound with a novel structure and efficacy by chemically modifying sodium pyruvate. The compound belongs to a type of pyruvate derivative. The compound can penetrate into the lung infection area and control the infection of lung biofilm. In addition, the present invention proves through a large number of experimental results that the prepared compound I has both anti-infection activity and biocompatibility, can significantly reduce the expression level of inflammatory factors, and has broad application prospects in the field of lung biofilm infection. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 is the hydrogen nuclear magnetic resonance spectrum of the intermediate 1 of the present invention; Figure 2 is the hydrogen nuclear magnetic resonance spectrum of intermediate 2 of the present invention; Figure 3 is the hydrogen nuclear magnetic resonance spectrum of intermediate 3 of the present invention; Figure 4 is the hydrogen nuclear magnetic resonance spectrum of compound I of the present invention; Figure 5 This is a graph showing the biocompatibility results of pyruvate of the present invention; Figure 6 This is a result diagram showing the effect of pyruvate on the inflammatory factor IL-1β of the present invention; Figure 7 This is a result diagram showing the effect of pyruvate on the inflammatory factor IL-6 in the present invention; Figure 8 This is a graph showing the effect of pyruvate on the inflammatory factor TNF-α. DETAILED DESCRIPTION

[0016] The technical solution of the present invention is further explained below in conjunction with specific embodiments, comparative examples, test examples and drawings.

[0017] In the following examples, comparative examples and test examples, the raw materials and preparation methods used are conventional materials and techniques in the art unless otherwise specified.

[0018] Example 1 The use of pyruvate in the preparation of a drug for treating pulmonary biofilm infection, wherein the pyruvate is compound I, and its structural formula is as follows: Compound I.

[0019] The preparation steps of compound I are as follows: (1) Weigh 4-chlorophenylhydrazine (0.11 mol) and 3-methyl-2-butanone (0.1 mol) and add them to 100 mL of anhydrous ethanol, then add 98% concentrated sulfuric acid (1.6 eq) to obtain a mixed solution, and reflux the mixed solution at 80°C for 1.5 h. After the reaction, adjust the pH value of the reaction solution to pH = 8.7 using 10wt% NaOH solution, extract with ethyl acetate, and distill the ethyl acetate phase under reduced pressure to obtain intermediate 1; The hydrogen nuclear magnetic resonance spectrum of the intermediate 1 is as follows: Figure 1 As shown, 1 HNMR (C 11 H 12 ClN, 400MHz, DMSO-d6)δ: 7.63 (s, 1H), 7.45 (d, 1H), 7.29(d, 1H), 1.95(s, 3H), 1.45(s, 6H); The above results confirmed that the obtained product was the target product.

[0020] (2) Add POCl3 (36.12 mmol) dropwise to a reaction bottle containing DMF (5.5 mL) under ice bath conditions and stir at 5°C for 5 min to obtain a mixed system. Then, add a solution of intermediate 1 (10.32 mmol) in DMF (5 mL) dropwise to the mixed system. Take the reaction bottle out of the ice bath and stir at 75°C for 4 h. After the reaction is completed, add ice water to the reaction solution, and then add NaOH aqueous solution to make it neutral. Filter the reaction solution to obtain a precipitate, dry it, and then recrystallize it with ethanol to obtain intermediate 2. The nuclear magnetic resonance hydrogen spectrum of the intermediate 2 is as follows Figure 2 As shown, 1 HNMR (C 13 H 12 ClNO2, 400MHz, DMSO-d6)δ: 12.71 (s, 1H), 10.15 (s, 2H), 7.45(d, 1H), 7.15(d, 1H), 7.02(d, 1H),1.77(s,6H); The above results confirmed that the obtained product was the target product.

[0021] (3) Weigh intermediate 2 (1 mmol), 3-cyanophenylhydrazine (1.2 mmol), glacial acetic acid (AcOH) (2.5 mmol) and ethanol (EtOH) (50 mL) into a reaction flask to obtain a mixed solution, which was refluxed for 18 h. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was filtered, washed with 50% ethanol aqueous solution (5 mL × 2) and dried. The crude product was purified by column chromatography using n-hexane and ethyl acetate (70:30~50:50 v / v) as the mobile phase to obtain intermediate 3. The nuclear magnetic resonance hydrogen spectrum of the intermediate 3 is as follows Figure 3 As shown, 1 HNMR (C 20 H 15 ClN4, 400MHz, DMSO-d6)δ: 8.49 (d, 1H), 8.17 (d, 1H), 7.92-7.40 (m, 6H), 7.26 (d, 1H), 1.41 (s, 6H); The above results confirmed that the obtained product was the target product.

[0022] (4) Weigh 3,4-dihydroxybenzoic acid (10 mmol) and sodium pyruvate (12 mmol) and dissolve them in 30 mL of ethanol / water mixed solvent (volume ratio of ethanol to water is 1:1) and add them to a 100 mL single-necked flask. Dissolve KOH (17.5 mmol) in 20 mL of ethanol and drip into the single-necked flask. Stir at room temperature for 6 h. After the reaction is completed, the reaction solution is concentrated under reduced pressure to obtain a crude product. The crude product is evenly dispersed with 40 mL of deionized water and washed with ethyl acetate (20 mL×3) 3 times. Add an appropriate amount of dilute hydrochloric acid to the aqueous phase until the pH value is 3; then, extract the aqueous phase three times with ethyl acetate (20 mL×3). After combining the organic phases, dry and concentrate to obtain the intermediate product 4. The above intermediate 4 1 HNMR (C 10 H8O5, 400MHz, DMSO-d6)δ: 12.59 (s, 1H), 9.50 (s, 2H), 7.72 (d, 1H), 7.09 (s, 1H), 6.84(d, 1H), 6.69 (m, 2H); The above results confirmed that the obtained product was the target product.

[0023] (5) Weigh the intermediate 4 (10 mmol) and 4-formylphenylboronic acid (12 mmol) and add them to a mixed solution of 100 mL of ethanol and water (volume ratio of 1:1), then adjust the pH value of the mixed solution to 8.2 with a 10 wt% NaOH aqueous solution, stir and react for 8 h. After the reaction is completed, extract, dry, concentrate and purify the reaction solution by column to obtain the intermediate 5. The above intermediate 5 1 HNMR (C 17 H 11 BO6, 400MHz, DMSO-d6)δ: 12.90 (s, 1H), 9.91 (s, 1H), 7.96-7.70 (m, 5H), 7.04 (d, 1H), 6.69-6.54 (m, 3H); The above results confirmed that the obtained product was the target product.

[0024] (6) Intermediate 3 (10 mmol) and intermediate 5 (12 mmol) were weighed and added to 100 mL of a mixed solution of ethanol and water (volume ratio 1:1). The mixed solution was then reacted under stirring for 13 h. After the reaction was completed, the reaction solution was concentrated and purified by column to obtain the final product, i.e., compound I.

[0025] The hydrogen nuclear magnetic resonance spectrum of the above compound I is as follows Figure 4 As shown, 1 HNMR (C 37 H 28 BClN4O5, 400MHz, DMSO-d6)δ: 12.59 (s, 1H), 8.18 (s, 1H), 7.80 (s, 1H), 7.72-7.64(m, 4H), 7.46-7.28(d, 5H), 7.04 (d, 1H), 6.69-6.25 (m, 4H), 5.35-5.28 (m, 2H), 2.15(s, 3H),1.45(s, 6H); The above results confirmed that the obtained product was the target product.

[0026] Example 2 The use of pyruvate in the preparation of a drug for treating lung biofilm infection, wherein the structural formula of the pyruvate is the same as that of compound I; the preparation steps of compound I are as follows: (1) Weigh 4-chlorophenylhydrazine (0.1 mol) and 3-methyl-2-butanone (0.1 mol) and add them to 100 mL of anhydrous ethanol, then add 98% concentrated sulfuric acid (1.2 eq) to obtain a mixed solution, and reflux the mixed solution at 80°C for 2 h. After the reaction, adjust the pH value of the reaction solution to pH = 8.5 with 10 wt% NaOH solution, extract with ethyl acetate, and distill the ethyl acetate phase under reduced pressure to obtain intermediate 1; (2) Add POCl3 (30.98 mmol) dropwise to a reaction bottle containing DMF (5.5 mL) under ice bath conditions and stir at 5°C for 5 min to obtain a mixed system. Then, add a solution of intermediate 1 (10.32 mmol) in DMF (5 mL) dropwise to the mixed system. Take the reaction bottle out of the ice bath and stir at 70°C for 5 h. After the reaction is completed, add ice water to the reaction solution, and then add NaOH aqueous solution to make it neutral. Filter the reaction solution to obtain a precipitate, dry it, and recrystallize it with ethanol to obtain intermediate 2. (3) Weigh intermediate 2 (1 mmol), 3-cyanophenylhydrazine (1.1 mmol), glacial acetic acid (AcOH) (2 mmol) and ethanol (EtOH) (50 mL) into a reaction flask to obtain a mixed solution, which was refluxed for 19 h. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was filtered, washed with 50% ethanol aqueous solution (5 mL × 2) and dried. The crude product was purified by column chromatography using n-hexane and ethyl acetate (70:30~50:50 v / v) as the mobile phase to obtain intermediate 3. (4) Weigh 3,4-dihydroxybenzoic acid (10 mmol) and sodium pyruvate (10 mmol) and dissolve them in 30 mL of ethanol / water mixed solvent (volume ratio of ethanol to water is 1:1) and add them to a 100 mL single-necked flask. Dissolve KOH (15 mmol) in 20 mL of anhydrous ethanol and drip into the single-necked flask. Stir at room temperature for 5 h. After the reaction is completed, the reaction solution is concentrated under reduced pressure to obtain a crude product. The crude product is evenly dispersed with 40 mL of deionized water and washed with ethyl acetate (20 mL×3) 3 times. Add an appropriate amount of dilute hydrochloric acid to the aqueous phase until the pH value is 3; then, extract the aqueous phase three times with ethyl acetate (20 mL×3), combine the organic phases, dry and concentrate to obtain the intermediate product 4; (5) Weigh the intermediate 4 (10 mmol) and 4-formylphenylboronic acid (10 mmol) and add them to 100 mL of a mixed solution of ethanol and water (volume ratio of 1:1). Then adjust the pH value of the mixed solution to 8.2 with a 10 wt% NaOH aqueous solution and stir for 7 h. After the reaction is completed, extract, dry, concentrate and purify the reaction solution by column to obtain the intermediate 5. (6) Intermediate 3 (10 mmol) and intermediate 5 (10 mmol) were weighed and added to a mixed solution of 100 mL of ethanol and water (volume ratio of 1:1), and then the mixed solution was reacted under stirring conditions for 12 h. After the reaction was completed, the reaction solution was concentrated, dried, and purified by column to obtain the final product, i.e., compound I.

[0027] Example 3 The use of pyruvate in the preparation of a drug for treating lung biofilm infection, wherein the structural formula of the pyruvate is the same as that of compound I; the preparation steps of compound I are as follows: (1) Weigh 4-chlorophenylhydrazine (0.12 mol) and 3-methyl-2-butanone (0.1 mol) and add them to 100 mL of anhydrous ethanol, then add 98% concentrated sulfuric acid (1.8 eq) to obtain a mixed solution, and reflux the mixed solution at 85°C for 1 h. After the reaction, adjust the pH value of the reaction solution to pH = 9.0 using 10wt% NaOH solution, extract with ethyl acetate, and distill the ethyl acetate phase under reduced pressure to obtain intermediate 1; (2) Add POCl3 (41.28 mmol) dropwise to a reaction bottle containing DMF (5.5 mL) under ice bath conditions and stir at 5°C for 5 min to obtain a mixed system. Then, add a solution of intermediate 1 (10.32 mmol) in DMF (5 mL) dropwise to the mixed system. Take the reaction bottle out of the ice bath and stir at 80°C for 3 h. After the reaction is completed, add ice water to the reaction solution, and then add NaOH aqueous solution to make it neutral. Filter the reaction solution to obtain a precipitate, dry it, and then recrystallize it with ethanol to obtain intermediate 2. (3) Weigh intermediate 2 (1 mmol), 3-cyanophenylhydrazine 1.3 mmol), glacial acetic acid (AcOH) (3 mmol) and ethanol (EtOH) (50 mL) into a reaction flask to obtain a mixed solution, which was refluxed for 20 h. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was filtered, washed with 50% ethanol aqueous solution (5 mL × 2) and dried. The crude product was purified by column chromatography using n-hexane and ethyl acetate (70:30~50:50 v / v) as the mobile phase to obtain intermediate 3. (4) Weigh 3,4-dihydroxybenzoic acid (10 mmol) and sodium pyruvate (15 mmol) and dissolve them in 30 mL of ethanol / water mixed solvent (volume ratio of ethanol to water is 1:1) and add them to a 100 mL single-necked flask. Dissolve KOH (20 mmol) in 20 mL of anhydrous ethanol and drip into the single-necked flask. Stir at room temperature for 7 h. After the reaction is completed, the reaction solution is concentrated under reduced pressure to obtain a crude product. The crude product is evenly dispersed with 40 mL of deionized water and washed with ethyl acetate (20 mL×3) 3 times. Add an appropriate amount of dilute hydrochloric acid to the aqueous phase until the pH value is 3; then, extract the aqueous phase three times with ethyl acetate (20 mL×3). After combining the organic phases, dry and concentrate to obtain the intermediate product 4. (5) Weigh the intermediate 4 (10 mmol) and 4-formylphenylboronic acid (15 mmol) and add them to a mixed solution of 100 mL of ethanol and water (volume ratio of 1:1), then adjust the pH value of the mixed solution to 8.2 with a 10 wt% NaOH aqueous solution, stir and react for 9 h. After the reaction is completed, extract, dry, concentrate and purify the reaction solution by column to obtain the intermediate 5. (6) Intermediate 3 (10 mmol) and intermediate 5 (15 mmol) were weighed and added to a mixed solution of 100 mL of ethanol and water (volume ratio of 1:1), and then the mixed solution was reacted under stirring conditions for 14 h. After the reaction was completed, the reaction solution was concentrated, dried, and purified by column to obtain the final product, i.e., compound I.

[0028] Comparative Example 1 Use of pyruvate in the preparation of a drug for treating pulmonary biofilm infection. The difference between Comparative Example 1 and Example 1 is that Compound I is replaced with sodium pyruvate.

[0029] Test Example 1 Biocompatibility test: Mouse fibroblast L929 cells were used to evaluate the biocompatibility of the product obtained in Example 1 of the present invention with L929 cells. First, fibroblast L929 cells in the logarithmic growth phase were resuspended in DMEM complete medium, and the cell density in the cell suspension was controlled to be 1×10 5 / mL, take 100 μL of the above cell suspension and inoculate it into a 96-well plate, and place it in an incubator at 37°C and 5% CO2 for 12 hours. Aspirate the culture medium, and add 200 μL of DMEM complete culture medium containing pyruvate obtained in Example 1 at a concentration of 30 μg / mL to the 96-well plate, and continue to culture under the above conditions for 24 hours. After waiting for the end of the culture, observe the cell morphology under a microscope. The present invention uses DMEM serum-free culture medium as the negative control group and DMEM complete culture medium supplemented with 5% phenol as the positive control group. The experimental results are as follows Figure 5 As shown, Figure 5 a is the cell compatibility diagram of the negative control group, Figure 5 b is the cell compatibility diagram of the experimental group, Figure 5 c is the cell compatibility diagram of the positive control group.

[0030] Depend on Figure 5 It can be seen that the cell morphology of the pyruvate obtained in Example 1 is similar to that of the negative control group, which belongs to the normal L929 cell morphology, indicating that the pyruvate obtained in Example 1 did not damage the growth morphology of mouse fibroblasts L929, while the cell morphology of the positive control group deteriorated. Therefore, the pyruvate obtained in Example 1 has good biocompatibility and can be used to prepare drugs for treating lung biofilm infections.

[0031] Test Example 2 2.1 Establishment of a mouse chronic lung infection model (1) Mouse grouping: 6-8 week old C57BL / 6 mice weighing 20-25 g were selected as experimental subjects. The mice were placed at 22±2°C and 55±5% relative humidity for one week for adaptive feeding. The mice were randomly divided into a blank control group, a model group, a medium dose group of Example 1, a high dose group of Example 1, a low dose group of Example 1, and a comparative example 1 group, with 10 mice in each group.

[0032] (2) Modeling: Pseudomonas aeruginosa (ATCC 27853) was used as the experimental strain and inoculated into LB broth to obtain a bacterial suspension. After 12 hours, the bacterial suspension was centrifuged and washed. Except for the blank control group, mice in other groups were given 50 μL of the cell suspension by airway infusion to make the number of bacteria in the mouse lungs 2×10 6 CFU.

[0033] (3) Administration process: Except for the model group, the products obtained in Example 1 and Comparative Example 1 were injected into the corresponding mice by tail vein injection on the 7th day of mouse infection, wherein the dosage of the low-dose group of Example 1 was 20 mg / kg, the dosage of the medium-dose group of Example 1 was 30 mg / kg, the dosage of the high-dose group of Example 1 was 40 mg / kg, and the dosage of the Comparative Example 1 group was 30 mg / kg, once a day, for a total of 3 times.

[0034] 2.2 Weight changes The body weight of mice was recorded before infection and on days 2, 4, 6, 8 and 10 after infection. The results are shown in Table 1, where the body weight on day 0 is the body weight before infection.

[0035] Table 1 From the experimental results in Table 1, it can be seen that the body weight of mice dropped sharply after infection, but after being treated with pyruvate in Example 1, their body weight tended to increase.

[0036] 2.3 Changes in the number of viable bacteria in the lungs of mice After 3 days of treatment, each mouse was intraperitoneally injected with 2% sodium pentobarbital solution for anesthesia, the thoracotomy was performed to expose the mouse trachea and lungs, the left main bronchus was ligated, and the bronchoalveoli were lavaged with 4°C phosphate buffer for 3 consecutive times to collect the mouse's bronchoalveolar lavage fluid (BALF). The mouse's lungs were then removed, homogenized and plated, and the number of viable bacteria was counted. The results are shown in Table 2.

[0037] Table 2 Number of viable bacteria in mouse lungs As shown in Table 2, compared with the model group, after 3 days of treatment with the pyruvate obtained in Example 1, the number of surviving bacteria in the lungs of mice was significantly reduced, showing excellent antibacterial efficacy. However, compared with the high, medium and low dose groups of Example 1, the antibacterial effect of Comparative Example 1 was poor. The above results show that the pyruvate obtained in Example 1 of the present invention exhibits excellent performance in treating biofilm infection in vivo.

[0038] 2.4 Changes in inflammatory factors The BALF obtained in 2.3 was centrifuged at 4000 rpm for 5 min to obtain the supernatant. The supernatant of each group of mice was tested for inflammatory factors IL-1β, IL-6 and TNF-α according to the instructions of the ELIAS kit. The results are as follows: Figure 6-8 shown.

[0039] Figure 6-8 The figures are the test results of the inflammatory factors IL-1β, IL-6 and TNF-α of mice in each group. As can be seen from the figure, compared with the model group, the contents of IL-1β, IL-6 and TNF-α in the mice in the high, medium and low dose groups of Example 1 were significantly reduced, while the decline trend of Comparative Example 1 was relatively slow, indicating that the pyruvate prepared by the present invention can well alleviate the lung inflammation caused by lung biofilm infection.

[0040] 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. Use of pyruvate in the preparation of a drug for treating pulmonary biofilm infection, characterized in that: The pyruvate is compound I, and its structural formula is shown below: Compound I.

2. The use of pyruvate according to claim 1 in the preparation of a drug for treating pulmonary biofilm infection, characterized in that: The preparation method of the pyruvate comprises the following steps: (1) Weighing 4-chlorophenylhydrazine and 3-methyl-2-butanone into ethanol, and then adding sulfuric acid to obtain a mixed solution, subjecting the mixed solution to reflux reaction. After the reaction is completed, adjusting the pH of the reaction solution to 8.5-9.0, and performing post-treatment to obtain intermediate 1; (2) Adding the DMF solution of intermediate 1 to the DMF solution of POCl3, heating for reaction, and after completion of the reaction, post-treatment to obtain intermediate 2; (3) adding intermediate 2, 3-cyanophenylhydrazine and glacial acetic acid to ethanol, and then reacting under reflux. After the reaction is completed, the reaction solution is cooled to room temperature, filtered, washed, dried and purified to obtain intermediate 3; (4) Dissolving 3,4-dihydroxybenzoic acid and sodium pyruvate in ethanol, then adding KOH ethanol solution thereto, stirring for reaction, and after completion of the reaction, post-treatment to obtain intermediate 4; (5) adding intermediate 4 and 4-formylphenylboronic acid to an aqueous solution of ethanol, adjusting the pH of the mixed solution to 8.2, stirring the mixture for reaction, and after the reaction is completed, purifying the reaction solution to obtain intermediate 5; (6) Add intermediate 3 and intermediate 5 to an aqueous solution of ethanol and stir to react. After the reaction is completed, concentrate, dry and purify the reaction solution to obtain the final product.

3. The use of pyruvate according to claim 2 in the preparation of a drug for treating pulmonary biofilm infection, characterized in that: In step (1), the molar ratio of 4-chlorophenylhydrazine, 3-methyl-2-butanone and sulfuric acid is (1-1.2):1:(1.2-1.8), and the reflux reaction time is 1-2 h.

4. The use of pyruvate according to claim 2 in the preparation of a drug for treating pulmonary biofilm infection, characterized in that: In the DMF solution of the intermediate 1 in step (2), the amount ratio of the intermediate 1 to DMF is 10.32 mmol: 5 mL; in the DMF solution of POCl3, the amount ratio of POCl3 to DMF is 36.12 mmol: 5.5 mL; the molar ratio of the intermediate 1 to POCl3 is 1: (3-4); the temperature of the heating reaction is 70-80°C, and the heating reaction time is 3-5 h.

5. The use of pyruvate according to claim 2 in the preparation of a drug for treating pulmonary biofilm infection, characterized in that: In step (3), the molar ratio of the intermediate 2, 3-cyanophenylhydrazine and glacial acetic acid is 1:(1.1-1.3):(2-3), and the reflux time is 18-20 h.

6. The use of pyruvate according to claim 2 in the preparation of a drug for treating pulmonary biofilm infection, characterized in that: In step (4), the molar ratio of 3,4-dihydroxybenzoic acid, sodium pyruvate and KOH is 1:(1-1.5):(1.5-2), and the reaction time is 5-7 h.

7. The use of pyruvate according to claim 2 in the preparation of a drug for treating pulmonary biofilm infection, characterized in that: In step (5), the molar ratio of the intermediate 4 to 4-formylphenylboronic acid is 1:(1-1.5), and the reaction time is 7-9 hours.

8. The use of pyruvate according to claim 2 in the preparation of a drug for treating pulmonary biofilm infection, characterized in that: In step (6), the molar ratio of intermediate 3 to intermediate 5 is 1:(1-1.5); and the reaction time is 12-14 h.

9. The use of pyruvate according to any one of claims 1 to 8 in the preparation of a drug for treating pulmonary biofilm infection, characterized in that: The pulmonary biofilm infection is a pulmonary biofilm infection caused by Pseudomonas aeruginosa.

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