Application of pyruvate pharmaceutical composition in the treatment of pulmonary fibrosis

Abstract

The present invention belongs to the field of biomedicine technology and specifically relates to the use of a pyruvate pharmaceutical composition in the treatment of pulmonary fibrosis. The pyruvate pharmaceutical composition comprises a proanthocyanidin derivative and pyruvate, wherein the mass ratio of the proanthocyanidin derivative to pyruvate is (2-4):1, and the structure of the proanthocyanidin derivative is: #imgabs0#. Experimental results show that the pyruvate pharmaceutical composition can effectively reduce the expression of inflammatory factors IL-6, IL-1β, and TNF-α in rat serum, and reduce the content of hydroxyproline in lung tissue, exerting an anti-pulmonary fibrosis effect and alleviating the progression of pulmonary fibrosis in rats.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and particularly relates to the application of a pyruvate pharmaceutical composition in the treatment of pulmonary fibrosis. Background Art

[0002] Pulmonary fibrosis is a serious interstitial lung disease characterized by the gradual replacement of normal lung tissue with fibrotic tissue, leading to a gradual loss of lung function. The progression of this disease is often irreversible and has a high mortality rate, with a median survival of only 2 to 5 years. The primary pathophysiological manifestations of pulmonary fibrosis are chronic alveolar epithelial cell damage and overexpression of profibrotic cytokines, which lead to the activation and proliferation of fibroblasts, in turn triggering the extensive deposition of extracellular matrix and the formation of fibrotic tissue.

[0003] Currently, the treatment of pulmonary fibrosis mainly includes drug therapy, non-drug therapy, treatment of comorbidities, and special management of acute exacerbations. In terms of drug therapy, anti-fibrotic drugs such as pirfenidone and nintedanib have been shown to delay the progression of the disease to a certain extent. Pirfenidone has anti-inflammatory, anti-fibrotic and antioxidant properties, while nintedanib is a multi-target tyrosine kinase inhibitor that can inhibit multiple growth factor receptors, thereby effectively delaying the decline of lung function in patients with pulmonary fibrosis, reducing the risk of acute exacerbations and prolonging survival. However, despite the certain efficacy of these drugs, the treatment of pulmonary fibrosis still faces many challenges, including drug side effects, continued disease progression, and a decline in patients' quality of life.

[0004] Sodium pyruvate, as the sodium salt form of pyruvate, is an important intermediate product in the human metabolic process and is involved in multiple physiological processes such as energy production, fat metabolism and antioxidant defense. As a naturally occurring compound, sodium pyruvate is widely used in the field of nutritional supplements due to its properties of promoting fat decomposition and improving energy utilization efficiency. In recent years, with the deepening of understanding of its biological activity, the potential role of sodium pyruvate in the treatment of various diseases has gradually attracted attention. In particular, in the regulation of inflammation and oxidative stress, sodium pyruvate has shown new application prospects, which provides new ideas for exploring its possibilities in the treatment of complex diseases such as pulmonary fibrosis. Therefore, the present invention proposes a pyruvate pharmaceutical composition for the treatment of pulmonary fibrosis, providing a new treatment strategy for patients with pulmonary fibrosis. Summary of the Invention

[0005] In order to overcome the deficiencies of the prior art, the primary purpose of the present invention is to provide a pyruvate pharmaceutical composition.

[0006] Another object of the present invention is to provide use of the above-mentioned pyruvate pharmaceutical composition in the preparation of a drug for treating pulmonary fibrosis.

[0007] The first object of the present invention is achieved through the following technical solutions:

[0008] A pyruvate pharmaceutical composition comprising the following parts by weight of a procyanidin derivative and pyruvate, wherein the mass ratio of the procyanidin derivative to the pyruvate is (2-4): 1;

[0009] The structural formula of the proanthocyanidin derivative is:

[0010] .

[0011] Preferably, the preparation process of the proanthocyanidin derivative is as follows:

[0012]

[0013] (1) Add methyl 3-formyl-4-hydroxybenzoate, 4-aminophenylboronic acid and NaHCO3 to solvent A, stir to react, and purify to obtain compound A;

[0014] (2) Compound A is added to DMF, and then 1,6-dibromohexane and cesium carbonate are added to heat the reaction, and compound B is obtained after purification;

[0015] (3) Compound B is added to acetonitrile, followed by N-methylpiperazine and K2CO3, and heated to react. Compound C is obtained after purification;

[0016] (4) Compound C is added to n-propanol, and then hydrazine hydrate is added and stirred to react, and compound D is obtained after purification;

[0017] (5) Add 4-acetylbenzaldehyde to methanol, then add pyruvic acid, and then add potassium hydroxide in batches. React at 30-40°C for 1-2 hours, cool to 0°C and continue to react for 10-12 hours. After purification, obtain compound E.

[0018] (6) Compound E and compound D are added to water, and then aluminum chloride is added, heated to react, and purified to obtain compound F;

[0019] (7) Add proanthocyanidin to NaHCO3 solution, then add compound F, stir to react, and purify to obtain proanthocyanidin derivatives.

[0020] Preferably, in step (1), the molar ratio of methyl 3-formyl-4-hydroxybenzoate, 4-aminophenylboronic acid, and NaHCO3 is 1: (1-1.5): (0.01-0.02); and the stirring reaction time is 12-16 h.

[0021] Preferably, in step (2), the molar ratio of compound A, 1,6-dibromohexane and cesium carbonate is 1:(1-2):(5-6); the heating reaction temperature is 60-70°C, and the time is 10-12 h.

[0022] Preferably, the molar ratio of compound B, N-methylpiperazine and K2CO3 in step (3) is 1:(1-2):(10-15); the heating reaction temperature is 80-90°C and the time is 3-5 h.

[0023] Preferably, the molar ratio of compound C to hydrazine hydrate in step (4) is 1:(2-3); and the stirring reaction time is 8-12 h.

[0024] Preferably, the molar ratio of 4-acetylbenzaldehyde, pyruvic acid and potassium hydroxide in step (5) is 1:(1-2):(1.5-2).

[0025] Preferably, the molar ratio of compound E, compound D and aluminum chloride in step (6) is 1:(1-2):(0.2-0.3); the heating reaction temperature is 50-70°C and the time is 6-10 h.

[0026] Preferably, the molar ratio of proanthocyanidins to compound F in step (7) is 1:(2-3); and the stirring reaction time is 2-4 h.

[0027] The second object of the present invention is achieved by the following technical solutions:

[0028] Use of the above pyruvate pharmaceutical composition in the preparation of a drug for treating pulmonary fibrosis.

[0029] The present invention has the following effects compared to the prior art:

[0030] 1. The present invention provides a pyruvate pharmaceutical composition comprising pyruvate and a proanthocyanidin derivative. Experimental results demonstrate that this pyruvate pharmaceutical composition effectively reduces the expression of inflammatory factors IL-6, IL-1β, and TNF-α in rat serum, and lowers the hydroxyproline content in lung tissue, exerting an anti-pulmonary fibrosis effect and alleviating the progression of pulmonary fibrosis in rats.

[0031] 2. The present invention also provides the use of the above-mentioned pyruvate pharmaceutical composition, which provides a new synthetic approach for preparing drugs for treating pulmonary fibrosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The following is a reaction flow chart for preparing proanthocyanidin derivatives according to the present invention;

[0033] Figure 2The weight changes of rats in each group in the test examples of the present invention;

[0034] Figure 3 is the IL-6 content in the serum of rats in each group in the test example of the present invention;

[0035] Figure 4 is the IL-1β content in the serum of rats in each group in the test example of the present invention;

[0036] Figure 5 is the TNF-α content in the serum of rats in each group in the test example of the present invention;

[0037] Figure 6 It is the content of HYP in the lung tissue of each group of rats in the test example of the present invention. DETAILED DESCRIPTION

[0038] The technical solution of the present invention will be further described below in conjunction with specific embodiments. However, it should be understood by those skilled in the art that the following examples are only intended to illustrate the present invention and should not be construed as limiting the present invention. Specific conditions not specified in the examples are to be followed according to conventional conditions or the conditions recommended by the manufacturer. All reagents or instruments used, unless otherwise specified, are conventional products obtained from commercial channels.

[0039] Example 1

[0040] A pyruvate pharmaceutical composition comprises a procyanidin derivative and sodium pyruvate, wherein the mass ratio of the procyanidin derivative to the sodium pyruvate is 2:1.

[0041] The preparation process of the proanthocyanidin derivatives is as follows Figure 1 As shown, the following steps are included:

[0042] (1) Methyl 3-formyl-4-hydroxybenzoate (CAS: 24589-99-9, 10 mmol), 4-aminophenylboronic acid (CAS: 89415-43-0, 12 mmol), and sodium bicarbonate (0.15 mmol) were added to 100 mL of ethanol and stirred for 14 h. The reaction mixture was concentrated and purified by column chromatography to obtain compound A (yield 62.6%).

[0043] Compound A 1 HNMR (C 15 H 14H-DENSO (BNO5, 400 MHz, d6-DMSO) δ11.13 (s, 1H), 8.90 (s, 1H), 8.17 (s, 1H), 8.03 (d, 1H), 7.76 (d, 2H), 7.15 (d, 2H), 7.04 (d, 1H), 4.23 (s, 2H), 3.90 (s, 3H); the above results confirmed that the target product was obtained.

[0044] (2) Compound A (10 mmol) was added to 100 mL of DMF, followed by the addition of 1,6-dibromohexane (CAS: 629-03-8, 15 mmol) and cesium carbonate (55 mmol), and the mixture was stirred at 65°C for 11 h. After cooling to room temperature, ethyl acetate and water were added to the reaction solution. The ethyl acetate phase was collected and saturated brine was added thereto to remove DMF. The organic phase was concentrated and purified by column chromatography to obtain compound B (yield 44.6%).

[0045] Compound B 1 HNMR (C 21 H 25 BBrNO5, 400MHz, d6-DMSO)δ8.90(s, 1H), 8.34(s, 1H),8.20(d, 1H), 7.76(d, 2H), 7.18(d, 1H), 7.15(d, 2H),4.22(s, 2H), 4.08(t, 2H),3.89(s, 3H), 3.54(t, 2H), 1.82-1.76(m, 2H), 1.45-1.42(m, 2H), 1.30-1.28(m,2H); MS(ESI)m / z=462.10[M+H + ], and found 462.10; the above results confirmed that the target product was obtained.

[0046] (3) Compound B (10 mmol) was added to 100 mL of acetonitrile, followed by the addition of N-methylpiperazine (CAS: 109-01-3, 15 mmol) and K2CO3 (120 mmol). The mixture was stirred at 85°C for 4 h. The reaction solution was concentrated, and ethyl acetate and water were added thereto. The ethyl acetate phase was collected and saturated brine was added thereto. The organic phase was concentrated and purified by column chromatography to obtain compound C (yield 51.8%).

[0047] Compound C 1 HNMR (C 26 H 36BN3O5, 400MHz, d6-DMSO)δ8.90(s, 1H), 8.34(s, 1H),8.20(d, 1H), 7.76(d, 2H), 7.18(d, 1H), 7.15(d, 2H),4.22(s, 2H), 4.08(t, 2H),3.89(s, 3H), 3.03(s, 2H), 2.30(s, 8H), 2.14(s, 3H), 1.78-1.75(m, 2H), 1.45-1.30(m, 6H); MS(ESI)m / z=482.27[M+H + ], and found 482.27; the above results confirmed that the target product was obtained.

[0048] (4) Compound C (10 mmol) was added to n-propanol, followed by hydrazine hydrate (25 mmol), and the mixture was refluxed for 10 h. The reaction mixture was rotary evaporated to remove the solvent and the remaining hydrazine hydrate to obtain a crude product. The crude product was purified by column chromatography to obtain compound D (40.2%).

[0049] Compound D 1 HNMR (C 26 H 36 BN5O4, 400MHz, d6-DMSO)δ9.81(s, 1H), 8.90(s, 1H),8.32(s, 1H), 8.18(d, 1H), 7.76(d, 2H), 7.26(d, 1H), 7.15(d, 2H), 4.49(s, 2H),4.22(s, 2H), 4.08(t, 2H), 3.03(s, 2H), 2.30(s, 8H), 2.14(s, 3H), 1.78-1.75(m,2H), 1.45-1.30(m, 6H); MS(ESI)m / z=482.29[M+H + ], and found 482.29; the above results confirmed that the target product was obtained.

[0050] (5) 4-Acetylbenzaldehyde (CAS: 3457-45-2, 10 mmol) was added to 100 mL of methanol, followed by pyruvic acid (15 mmol) and potassium hydroxide (12 mmol). After stirring for 30 min, potassium hydroxide (6 mmol) was added and the mixture was reacted at 35 °C for 1.5 h. The mixture was then cooled to 0 °C and reacted for 11 h. After the reaction was completed, the product was filtered and washed twice with cold methanol and cold ether. The product was then dried under vacuum to obtain compound E (yield 76.8%).

[0051] Compound E 1 HNMR (C 12 H 10 O4, 400MHz, d6-DMSO)δ12.56(s, 1H), 7.87(d, 2H),7.71(d, 1H), 7.46(d, 2H), 6.68(d, 1H), 2.51 (s, 3H); MS(ESI)m / z=219.06[M+H + ], and found 219.06; the above results confirmed that the target product was obtained.

[0052] (6) Compound E (10 mmol) and compound D (15 mmol) were added to 50 mL of deionized water, and then aluminum chloride hexahydrate (2.5 mmol) was added. The mixture was reacted at 60 °C for 8 h. The product was collected by filtration, washed with deionized water, and dried to obtain compound F (yield 48.9%).

[0053] Compound F 1 HNMR (C 37 H 44 BN5O7, 400MHz, d6-DMSO)δ12.56(s, 1H), 10.46(s, 1H),8.90(s, 1H), 8.32(s, 1H), 8.18(d, 1H), 7.76-7.75(m, 4H), 7.71(d, 1H), 7.47(d,2H), 7.26(d, 1H), 7.15(d, 2H), 6.68(d, 1H), 4.22(s, 2H), 4.08(t, 2H), 3.03(s,2H), 2.45(s, 3H), 2.30(s, 8H), 2.14(s, 3H), 1.78-1.75(m, 2H), 1.45-1.30(m,6H); MS(ESI)m / z=682.33[M+H + ], and found 682.33; the above results confirmed that the target product was obtained.

[0054] (7) Proanthocyanidin (CAS: 4852-22-6, 10 mmol) was added to 100 mL of 0.1 M NaHCO3 solution, and then compound F (25 mmol) was added. The mixture was stirred for 3 h. After the reaction was completed, the reaction solution was added to a dialysis bag (molecular weight cutoff 500 Da) and dialyzed for 48 h to obtain a proanthocyanidin derivative (yield 36.7%).

[0055] Proanthocyanidin derivatives 1 HNMR (C 104 H106 B2N 10 O 23 , 400MHz, d6-DMSO)δ12.56(s, 2H),10.46 (s, 2H), 10.29(s, 2H), 9.68(s, 2H), 8.95(s, 1H), 8.90(s, 1H), 8.32(s,2H), 8.18(d, 2H), 7.76-7.75(m, 8H), 7.71(d, 2H), 7.47(d, 4H), 7.26(d, 2H),7.15(d, 4H), 6.93(s, 1H), 6.82-6.80(m, 2H), 6.67-6.65(m, 6H), 5.94-5.89(m,4H), 5.08-5.06(m, 2H), 5.02(s, 1H), 4.78(d, 1H), 4.57-4.55(m, 1H), 4.08(t,4H), 3.03(s, 4H), 2.80-2.78(m, 1H), 2.55-2.53(m, 1H),2.45(s, 6H), 2.30(s,16H), 2.14(s, 6H), 1.78-1.75(m, 4H), 1.45-1.30(m, 12H); MS(ESI)m / z=1885.76 [M+H + ], and found 1885.76; the above results confirmed that the target product was obtained.

[0056] Example 2

[0057] A pyruvate pharmaceutical composition comprises a procyanidin derivative and sodium pyruvate, wherein the mass ratio of the procyanidin derivative to the sodium pyruvate is 4:1.

[0058] The preparation process of the proanthocyanidin derivatives is as follows Figure 1 As shown, the following steps are included:

[0059] (1) Methyl 3-formyl-4-hydroxybenzoate (10 mmol), 4-aminophenylboronic acid (10 mmol) and sodium bicarbonate (0.1 mmol) were added to 100 mL of ethanol and stirred for 12 h. The reaction mixture was concentrated and purified by column chromatography to obtain compound A (yield 41.8%). 1 HNMR is consistent with Example 1;

[0060] (2) Compound A (10 mmol) was added to 100 mL of DMF, followed by the addition of 1,6-dibromohexane (10 mmol) and cesium carbonate (50 mmol), and the mixture was stirred at 60°C for 12 h. After cooling to room temperature, ethyl acetate and water were added to the reaction solution, the ethyl acetate phase was collected and saturated brine was added thereto to remove DMF, the organic phase was concentrated and purified by column chromatography to obtain compound B (yield 43.3%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1;

[0061] (3) Compound B (10 mmol) was added to 100 mL of acetonitrile, followed by N-methylpiperazine (10 mmol) and K2CO3 (100 mmol). The mixture was stirred at 80°C for 5 h. The reaction solution was concentrated and ethyl acetate and water were added thereto. The ethyl acetate phase was collected and then saturated brine was added thereto. The organic phase was concentrated and purified by column chromatography to obtain compound C (yield 49.6%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1;

[0062] (4) Compound C (10 mmol) was added to n-propanol, and then hydrazine hydrate (20 mmol) was added, and the mixture was refluxed for 8 h. The reaction mixture was rotary evaporated to remove the solvent and the remaining hydrazine hydrate to obtain a crude product. The crude product was purified by column chromatography to obtain compound D (yield 39.7%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1;

[0063] (5) 4-Acetylbenzaldehyde (10 mmol) was added to 100 mL of methanol, followed by pyruvic acid (10 mmol) and potassium hydroxide (10 mmol). After stirring for 30 min, potassium hydroxide (5 mmol) was added and the mixture was reacted at 30 °C for 2 h. The mixture was then cooled to 0 °C and reacted for 10 h. After the reaction was completed, the product was filtered and washed twice with cold methanol and cold ether. The product was then vacuum dried to obtain compound E (yield 76.0%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1;

[0064] (6) Compound E (10 mmol) and compound D (10 mmol) were added to 50 mL of deionized water, and then aluminum chloride hexahydrate (2 mmol) was added and reacted at 50 °C for 10 h. The product was collected by filtration, washed with deionized water, and dried to obtain compound F (yield 46.1%). 1HNMR and MS (ESI) m / z were consistent with those in Example 1;

[0065] (7) Proanthocyanidin (10 mmol) was added to 100 mL of 0.1 M NaHCO3 solution, and then compound F (20 mmol) was added and stirred for 2 h. After the reaction was completed, the reaction solution was added to a dialysis bag (molecular weight cut-off 500 Da) and dialyzed for 48 h to obtain proanthocyanidin derivatives (yield 34.8%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1.

[0066] Example 3

[0067] A pyruvate pharmaceutical composition comprises a procyanidin derivative and sodium pyruvate, wherein the mass ratio of the procyanidin derivative to the sodium pyruvate is 3:1.

[0068] The preparation process of the proanthocyanidin derivatives is as follows Figure 1 As shown, the following steps are included:

[0069] (1) Methyl 3-formyl-4-hydroxybenzoate (10 mmol), 4-aminophenylboronic acid (15 mmol) and sodium bicarbonate (0.2 mmol) were added to 100 mL of ethanol and stirred for 16 h. The reaction mixture was concentrated and purified by column chromatography to obtain compound A (yield 43.1%). 1 HNMR was consistent with that of Example 1.

[0070] (2) Compound A (10 mmol) was added to 100 mL of DMF, followed by the addition of 1,6-dibromohexane (20 mmol) and cesium carbonate (60 mmol), and the mixture was stirred at 70°C for 10 h. After cooling to room temperature, ethyl acetate and water were added to the reaction solution, the ethyl acetate phase was collected and saturated brine was added thereto to remove DMF, the organic phase was concentrated and purified by column chromatography to obtain compound B (yield 45.2%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1.

[0071] (3) Compound B (10 mmol) was added to 100 mL of acetonitrile, followed by N-methylpiperazine (20 mmol) and K2CO3 (150 mmol), and the mixture was stirred at 90°C for 3 h. The reaction solution was concentrated, and ethyl acetate and water were added thereto. The ethyl acetate phase was collected and then saturated brine was added thereto. The organic phase was concentrated and purified by column chromatography to obtain compound C (yield 50.4%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1.

[0072] (4) Compound C (10 mmol) was added to n-propanol, and then hydrazine hydrate (30 mmol) was added, and the mixture was refluxed for 12 h. The reaction mixture was rotary evaporated to remove the solvent and the remaining hydrazine hydrate to obtain a crude product. The crude product was purified by column chromatography to obtain compound D (yield 36.3%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1.

[0073] (5) 4-Acetylbenzaldehyde (10 mmol) was added to 100 mL of methanol, followed by pyruvic acid (20 mmol) and potassium hydroxide (10 mmol). After stirring for 30 min, potassium hydroxide (10 mmol) was added again. The mixture was reacted at 40 °C for 1 h, and then cooled to 0 °C for 12 h. After the reaction was completed, the product was filtered and washed twice with cold methanol and cold ether. The product was then vacuum dried to obtain compound E (yield 77.0%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1.

[0074] (6) Compound E (10 mmol) and compound D (20 mmol) were added to 50 mL of deionized water, and then aluminum chloride hexahydrate (3 mmol) was added and reacted at 70 °C for 6 h. The product was collected by filtration, washed with deionized water, and dried to obtain compound F (yield 46.2%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1.

[0075] (7) Proanthocyanidin (10 mmol) was added to 100 mL of 0.1 M NaHCO3 solution, and then compound F (30 mmol) was added and stirred for 4 h. After the reaction was completed, the reaction solution was added to a dialysis bag (molecular weight cut-off 500 Da) and dialyzed for 48 h to obtain proanthocyanidin derivatives (yield 38.1%). 1 HNMR and MS (ESI) m / z were consistent with those in Example 1.

[0076] Comparative Example 1

[0077] The difference between the comparative example and Example 1 is that the proanthocyanidin derivative is replaced by proanthocyanidin.

[0078] Comparative Example 2

[0079] The difference between Comparative Example 2 and Example 1 is that the proanthocyanidin derivative is omitted.

[0080] Test example

[0081] 1. Experimental Animals, Grouping, and Modeling

[0082] Eighty male SPF Sprague-Dawley rats, weighing 200-220 g, were randomly divided into a model group, Example 1-3 groups, Comparative Example 1-2 groups, a positive control group, and a blank control group, with 10 rats in each group. The rats were acclimated for 3 days. Anesthesia was achieved by intraperitoneal injection of 4% chloral hydrate (10 mL / kg). After anesthesia, the rats were immobilized, the neck hair removed, and a longitudinal incision made to expose the trachea. In all groups except the blank control group, a syringe was used to penetrate the trachea 1 cm toward the cardiac end through the gap between the two tracheal cartilage rings. After withdrawing without resistance, bleomycin (5 mg / kg) was injected into the trachea of ​​the other groups. The blank control group was injected with an equal volume of saline. Immediately after injection, the incisions were sutured and disinfected. The rat board was then rotated upright for 3 minutes to ensure even distribution of the drug in the lungs. The rats were returned to a dry, clean cage for rest. After waking, the rats were housed and observed as normal.

[0083] 2. Dosage regimen

[0084] Positive control group: prednisone acetate was given from the day after modeling, at a dose of 5 mg / kg, once a day by gavage for 28 consecutive days;

[0085] Example 1-3 group and Comparative Example 1-2 group: Starting from the day after modeling, the drug of Example 1-3 and Comparative Example 1-2 groups was administered at a dose of 0.8 mg / kg (calculated as sodium pyruvate) by gavage once a day for 28 consecutive days;

[0086] Blank control group and model group: administered with equal volume of normal saline.

[0087] 3. Detection indicators and methods

[0088] 3.1 General Observation

[0089] After modeling, the daily eating status, fur color changes, respiration and external appearance of the rats in each group were observed and recorded; the weight changes of the rats in each group were observed, and the weight of the rats on days 0, 7, 14, 21 and 28 were recorded. The results are shown in Figure 2 .

[0090] 3.2 Pulmonary function tests

[0091] 24 h after the last administration, rats in each group were anesthetized by intraperitoneal injection of 50 mg / kg of Zoltrate. After anesthesia, tracheal intubation was performed and the rats were connected to a small animal pulmonary function analyzer to measure the pulmonary function indicators of the rats in each group: inspiratory capacity (IC), dynamic lung compliance (Cdyn), forced expiratory flow at 10% FVC (FEF) and respiratory rate. 50% ), the results are shown in Table 1.

[0092] 3.3 Serum levels of inflammatory factors

[0093] After the pulmonary function test, the rats in each group were anesthetized with 10% chloral hydrate, and blood was collected from the abdominal aorta. After standing at room temperature for 30 minutes, the blood was centrifuged (4500 rpm, 5 minutes, 4°C) to separate the serum. The ELISA kit instructions were strictly followed to detect the levels of IL-6, IL-1β, and TNF-α in the serum. The experimental results are the average values ​​of each group of rats. Figure 3-5 .

[0094] 3.4 Lung coefficient and HYP content in lung tissue

[0095] After blood was drawn, the rats in each group were killed, and the lung tissues were removed and weighed. The lung coefficient was calculated as follows: lung coefficient = lung wet weight (mg) / body weight (mg) × 100%. The hydroxyproline content in the lung tissues was detected by enzyme-linked immunosorbent assay (ELISA). The results are shown in Tables 2 and Figure 6 .

[0096] 4. Results Analysis

[0097] 4.1 Effects on rat status and body weight

[0098] The rats in the blank control group were in good mental state, with shiny fur, normal diet and drinking water, quick reactions, gradual weight gain, and normal breathing. The rats in the model group were listless within one week of modeling, with less activity, less diet and drinking water, slow reactions, dull fur, varying degrees of accelerated breathing, and no obvious weight gain. As the number of days increased, the symptoms progressively worsened, and some of the rats experienced hair loss. The rats in each treatment group were significantly better than those in the model group, and the rats in Examples 1-3 of the present invention and the positive control group were closer to the normal group.

[0099] The body weight of rats was recorded on days 0, 7, 14, 21 and 28. Figure 2 .Depend on Figure 2 It can be seen that compared with the blank group, the model group rats showed the most significant weight loss. Compared with the model group rats, the weight of the rats in Examples 1-3 groups showed a significant increase, which was better than that of the comparative example 1-2 groups and comparable to the positive control group. This shows that the pyruvate pharmaceutical composition prepared in Examples 1-3 can improve the physical fitness of pulmonary fibrosis model rats and significantly alleviate the effects of pulmonary fibrosis on rats.

[0100] 4.2 Effects on lung function in rats with pulmonary fibrosis

[0101] Table 1 Pulmonary function indicators of rats in each group

[0102]

[0103] As can be seen from Table 1, compared with the blank control group, the deep inspiration volume, dynamic lung compliance and FEF of the rats in the model group were 50% Compared with the model group, the deep inspiration volume, dynamic lung compliance and FEF of rats in Example 1-3 groups were significantly reduced. 50% The deep inspiratory capacity, dynamic lung compliance and FEF of rats in groups 1 and 2 were significantly increased, and there was no significant difference compared with the positive control group. 50% The above results show that the pyruvate pharmaceutical composition of the present invention can improve the respiratory function of rats with pulmonary fibrosis.

[0104] 4.3 Serum levels of inflammatory factors

[0105] The levels of IL-6, IL-1β, and TNF-α in the serum of rats in each group can be found in Figure 3 、 Figure 4 and Figure 5 .

[0106] from Figure 3 、 Figure 4 and Figure 5 It can be seen that compared with the blank control group, the levels of IL-6, IL-1β, and TNF-α in the serum of the model group rats were significantly increased. Compared with the model group, the levels of IL-6, IL-1β, and TNF-α in the serum of the positive control group, the Example 1-3 group, and the Comparative Example 1-2 group rats were all reduced, among which the reduction in IL-6, IL-1β, and TNF-α in the serum of the Example 1-3 group rats was comparable to that of the positive control group, and was significantly more than that of the Comparative Example 1-2 group. The above results show that the pyruvate drug combination of the present invention can effectively reduce the expression of inflammatory factors IL-6, IL-1β, and TNF-α in rat serum, thereby exerting an anti-pulmonary fibrosis effect.

[0107] 4.4 Effects on the lung coefficient and hydroxyproline content in lung tissue of rats with pulmonary fibrosis

[0108] Table 2 Lung coefficients of rats in each group

[0109]

[0110] As shown in Table 2, the lung organ coefficient of the model group rats was significantly higher than that of the blank control group, while the lung coefficients of the positive control group and Example 1-3 groups were significantly lower than those of the model group, and the lung coefficients of the comparative example 1-2 groups were lower than those of the model group.

[0111] Depend on Figure 6As can be seen, compared with the blank control group, the hydroxyproline content in the lung tissue of the model group was significantly increased. Compared with the model group, the hydroxyproline content in the lung tissue of rats in Examples 1-3 was reduced, with the effect comparable to that of the positive control group and superior to that of Comparative Examples 1-2. These results demonstrate that the pyruvate pharmaceutical composition of the present invention can effectively alleviate the progression of pulmonary fibrosis in rats.

[0112] Finally, it should be noted that the above embodiments are intended only to illustrate the technical solutions of the present invention and are not intended to limit the same. The basic principles and main features of the present invention have been described above using specific implementation schemes. Modifications or substitutions may be made based on the present invention, but such modifications or substitutions do not detract from the essence of the corresponding technical solutions from the scope of protection claimed by the present invention.

Claims

1. A pyruvate pharmaceutical composition, characterized in that: The invention comprises a proanthocyanidin derivative and pyruvate, wherein the mass ratio of the proanthocyanidin derivative to the pyruvate is (2-4):1; the pyruvate is sodium pyruvate; The structural formula of the proanthocyanidin derivative is: 。 2. The pyruvate pharmaceutical composition according to claim 1, characterized in that The preparation process of the proanthocyanidin derivative is as follows: (1) Add methyl 3-formyl-4-hydroxybenzoate, 4-aminophenylboronic acid and NaHCO3 to ethanol, stir and react, and purify to obtain compound A; (2) Compound A is added to DMF, and then 1,6-dibromohexane and cesium carbonate are added to heat the reaction, and compound B is obtained after purification; (3) Compound B is added to acetonitrile, followed by N-methylpiperazine and K2CO3, and heated to react. Compound C is obtained after purification; (4) Compound C is added to n-propanol, and then hydrazine hydrate is added and stirred to react, and compound D is obtained after purification; (5) Add 4-acetylbenzaldehyde to methanol, then add pyruvic acid, and then add potassium hydroxide in batches. React at 30-40°C for 1-2 hours, cool to 0°C and continue to react for 10-12 hours. After purification, obtain compound E. (6) Compound E and compound D are added to water, and then aluminum chloride is added, heated to react, and purified to obtain compound F; (7) Add proanthocyanidin to NaHCO3 solution, then add compound F, stir to react, and purify to obtain proanthocyanidin derivatives.

3. The pyruvate pharmaceutical composition according to claim 2, characterized in that The molar ratio of methyl 3-formyl-4-hydroxybenzoate, 4-aminophenylboronic acid and NaHCO3 in step (1) is 1: (1-1.5): (0.01-0.02); the stirring reaction time is 12-16 h.

4. The pyruvate pharmaceutical composition according to claim 2, characterized in that In step (2), the molar ratio of compound A, 1,6-dibromohexane and cesium carbonate is 1:(1-2):(5-6); the heating reaction temperature is 60-70°C and the time is 10-12 h.

5. The pyruvate pharmaceutical composition according to claim 2, characterized in that In step (3), the molar ratio of compound B, N-methylpiperazine and K2CO3 is 1:(1-2):(10-15); the heating reaction temperature is 80-90°C and the time is 3-5 h.

6. The pyruvate pharmaceutical composition according to claim 2, characterized in that In step (4), the molar ratio of compound C to hydrazine hydrate is 1:(2-3); and the stirring reaction time is 8-12 h.

7. The pyruvate pharmaceutical composition according to claim 2, characterized in that The molar ratio of 4-acetylbenzaldehyde, pyruvic acid and potassium hydroxide in step (5) is 1:(1-2):(1.5-2).

8. The pyruvate pharmaceutical composition according to claim 2, characterized in that In step (6), the molar ratio of compound E, compound D and aluminum chloride is 1:(1-2):(0.2-0.3); the heating reaction temperature is 50-70°C and the time is 6-10 h.

9. The pyruvate pharmaceutical composition according to claim 2, characterized in that The molar ratio of proanthocyanidin to compound F in step (7) is 1:(2-3); the stirring reaction time is 2-4 h.

10. Use of the pyruvate pharmaceutical composition according to any one of claims 1 to 9 in the preparation of a drug for treating pulmonary fibrosis.

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