Medicine for treating chronic obstructive pulmonary disease

Abstract

The invention discloses a medicine for treating chronic obstructive pulmonary disease, and belongs to the technical field of medicine. The active ingredient of the medicine is syringin. Experimental results show that syringin can significantly relieve airflow blockage, inhibit airway inflammation and improve lung tissue injury. More importantly, the invention proves that the syringin and aminophylline are combined for use to achieve a synergistic effect. According to the combined treatment scheme, the curative effect can be remarkably improved, and the dosage and toxic and side effects of aminophylline can be remarkably reduced. Therefore, the invention provides a more efficient and safer innovative medicine and a combined medication strategy thereof for COPD treatment, and has important clinical application value.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a therapeutic drug for chronic obstructive pulmonary disease. BACKGROUND

[0002] Chronic obstructive pulmonary disease (COPD) is a common chronic respiratory disease, mainly characterized by persistent airflow limitation, usually progressive development, and accompanied by abnormal inflammatory response of airway and lung tissue. The disease is mainly caused by long-term smoking, air pollution, occupational dust exposure or genetic factors (such as alpha 1-antitrypsin deficiency). Clinical manifestations include chronic cough, sputum, dyspnea and wheezing, and severe cases can lead to respiratory failure, heart and lung failure and even death.

[0003] In the prior art, the treatment of COPD mainly relies on drug intervention and non-drug support. Drug treatment includes bronchodilators, inhaled glucocorticoids and phosphodiesterase inhibitors, which aim to relieve symptoms, improve lung function and reduce acute exacerbation. Non-drug treatment includes smoking cessation, pulmonary rehabilitation training, oxygen therapy and vaccination (such as influenza and pneumonia vaccine) to prevent infection and improve quality of life. However, these existing treatments have significant limitations, for example, many patients have no response or gradually lose response to drugs such as beta-adrenergic agonists. Systemic treatment with glucocorticoids can cause side effects such as osteoporosis, cardiovascular risk, etc. Therefore, it is of great significance to develop new drugs that are safer and more effective for the treatment of chronic obstructive pulmonary disease.

[0004] Syringin is a glycoside compound extracted from plants such as Eleutherococcus senticosus. Its molecular formula is C 17 H 24 O9, molecular weight is 372.37, CAS number is 118-34-3, and the structural formula is as follows:

[0005] Current literature shows that syringin has anti-inflammatory, antioxidant, immunomodulatory and other functions, but the research on syringin in COPD is limited, and its function and effect are not known. SUMMARY

[0006] The purpose of the present application is to provide a new therapeutic drug for chronic obstructive pulmonary disease.

[0007] To achieve the above purpose, the present application provides the following technical solutions: Firstly, the present application provides the use of syringin in the preparation of a drug for treating chronic obstructive pulmonary disease, wherein the drug comprises syringin as an active ingredient and a pharmaceutically acceptable carrier or excipient.

[0008] Preferably, the molecular formula of the syringin is C 17 H 24 O9, with a molecular weight of 372.37 and a CAS number of 118-34-3. Preferably, the drug is used for treating chronic obstructive pulmonary disease by relieving airflow obstruction, inhibiting airway inflammation and / or improving lung tissue pathological damage.

[0009] Preferably, the drug is prepared to be administered at a dose of 20 mg / kg / day to 40 mg / kg / day of syringin content.

[0010] Secondly, the present application provides a drug for treating chronic obstructive pulmonary disease, which is composed of an active ingredient of syringin and a pharmaceutically acceptable carrier or excipient. The drug is prepared to be administered at a dose of 20 mg / kg / day to 40 mg / kg / day of syringin content.

[0011] Preferably, the molecular formula of the syringin is C 17 H 24 O9, with a molecular weight of 372.37 and a CAS number of 118-34-3. The drug is used for treating chronic obstructive pulmonary disease by relieving airflow obstruction, inhibiting airway inflammation and / or improving lung tissue pathological damage.

[0012] In addition, the present application provides the use of a composition in the preparation of a combined drug for treating chronic obstructive pulmonary disease, wherein the composition is composed of syringin and aminophylline. In the composition, the mass ratio of syringin to aminophylline is 2:1.

[0013] Preferably, the drug is prepared to be administered at a dose of 20 mg / kg / day of syringin content and 10 mg / kg / day of aminophylline content.

[0014] Finally, the present application provides a combined therapeutic pharmaceutical composition for treating chronic obstructive pulmonary disease, which is composed of syringin and aminophylline. In the combined therapeutic pharmaceutical composition, the mass ratio of syringin to aminophylline is 2:1.

[0015] Preferably, the combined therapeutic pharmaceutical composition is prepared to be administered at a dose of 20 mg / kg / day of syringin content and 10 mg / kg / day of aminophylline content.

[0016] The present application has the beneficial effect that: The present application has the beneficial effect that the natural compound syringin is first discovered and disclosed to have a brand-new use in treating chronic obstructive pulmonary disease (COPD). As an active ingredient, syringin can significantly relieve airflow obstruction, effectively inhibit airway inflammatory response, and improve pathological damage of lung tissue, thus providing a safe and effective innovative approach for COPD treatment.

[0017] In addition, it is unexpectedly found that syringin can produce a significant synergistic effect when used in combination with the traditional COPD drug aminophylline. This combination therapy far exceeds single-drug therapy in terms of efficacy, and more importantly, it can achieve an ideal therapeutic effect at a lower dose, thereby greatly reducing the dosage of aminophylline and its potential toxic side effects, providing a more efficient and safer new choice for combination therapy in the clinic, and having important clinical application prospects. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 The body weight changes of mice in each experimental group at week 0 (before the experiment began), week 8 (after COPD modeling was completed), and week 16 (after 8 weeks of treatment); The horizontal axis represents time points (week 0, week 8, week 16), and the vertical axis represents body weight; Figure 2 The ratio of forced expiratory volume at 0.05 seconds (FEV0.05) to forced vital capacity (FVC) (FEV0.05 / FVC) of mice in each experimental group after 8 weeks of treatment, as determined by the eSpira FM System lung function detection system; The horizontal axis represents experimental groups, and the vertical axis represents the FEV0.05 / FVC ratio; Figure 3 The detection results of IL-6 levels in bronchoalveolar lavage fluid (BALF) of mice in each experimental group; The horizontal axis represents experimental groups, and the vertical axis represents IL-6 concentration; Figure 4 The microscope observation results of HE-stained lung tissue sections of mice in each experimental group; Figure 5 The body weight changes of mice in each group in the combination therapy experiment at week 0, week 8, and week 16; The horizontal axis represents time points, and the vertical axis represents body weight; Figure 6 The FEV0.05 / FVC ratio of mice in each group in the combination therapy experiment; The horizontal axis represents experimental groups, and the vertical axis represents the FEV0.05 / FVC ratio; Figure 7The results of the detection of IL-6 levels in the bronchoalveolar lavage fluid (BALF) of each group of mice in the combination therapy experiment are shown in the following table: The horizontal axis represents the experimental group, and the vertical axis represents the IL-6 concentration. DETAILED DESCRIPTION

[0019] Example 1 (1) Animal preparation and grouping The purchased 6-8 week old C57BL / 6J mice were placed in a SPF environment (temperature 22±2℃, humidity 50±10%, 12 h light / dark cycle) and adaptively fed for 1 week; (2) Random grouping according to body weight and gender (15 mice per group): Normal control group (Control): exposed to filtered air, intranasal instillation of equal volume of normal saline, after modeling, oral gavage of normal saline; COPD model group (CS+KP): cigarette smoke exposure + Klebsiella pneumoniae instillation, after modeling, oral gavage of normal saline; Low-dose syringin group: after modeling, oral gavage of 20 mg / kg syringin once a day for 8 weeks of continuous treatment; High-dose syringin group: after modeling, oral gavage of 40 mg / kg syringin once a day for 8 weeks of continuous treatment.

[0020] (3) Preparation of Klebsiella pneumoniae bacterial solution Inoculate Klebsiella pneumoniae into LB medium and cultivate at 37℃ with shaking until the logarithmic growth phase; Centrifuge to collect the bacterial cells, resuspend with sterile normal saline, and adjust the concentration to 5×10 6 CFU / mL; (4) Klebsiella pneumoniae infection After mild anesthesia of the mice with sodium pentobarbital (50 mg / kg, oral gavage), fix the mice in a supine position, and slowly instill 20 μL of bacterial solution (5×10 6 CFU / mL) into both nostrils (10 μL per side, ensuring inhalation into the lungs) using a micropipette; Instillation frequency: once a week (0, 7, 14, 21, 28, 35, 42, 49 days), a total of 8 times; Control group: instill equal volume of sterile normal saline; (5) Cigarette smoke exposure Place the mice in a smoke chamber and expose them to cigarette smoke, ensuring that the smoke concentration is stable at 3000±500 ppm; Exposure time: twice a day (8:00 am and 16:00 pm), each time for 40 minutes, for 8 weeks (7 days a week); The control group was placed in the same device and exposed to filtered air.

[0021] (6) Drug administration Drug administration time: After the modeling was completed, drug administration was started at week 9, and continuous administration was performed for 8 weeks. Drug administration method: Oral gavage, using a 1 mL syringe and a gavage needle (22G, curved).

[0022] Blank control group and model group: 0.9% sodium chloride solution (normal saline) was administered orally, at a dose of 0.2 mL / day / animal; Low-dose syringin group: Oral gavage of 20 mg / kg syringin (dissolved in 0.9% sodium chloride); High-dose syringin group: Oral gavage of 40 mg / kg syringin (dissolved in 0.9% sodium chloride); Drug administration frequency: Once a day (9:00 am), for 8 weeks (56 days); The health status and adverse reactions of the mice were detected.

[0023] (7) Evaluation of mouse body weight The body weights of the mice in each group were measured at weeks 0, 8, and 16, and statistical graphs were drawn. Figure 1 ).

[0024] In the mice at week 0, Figure 1 the body weight of the control group was 22.42±0.89 g, the body weight of the COPD model group was 21.99±1.08 g, the body weight of the low-dose syringin group was 22.24±0.90 g, and the body weight of the high-dose syringin group was 21.73±0.94 g.

[0025] In the mice at week 8, the body weight of the control group was 33.39±1.00 g, the body weight of the COPD model group was 26.46±0.91 g, the body weight of the low-dose syringin group was 26.41±1.11 g, and the body weight of the high-dose syringin group was 26.56±1.13 g.

[0026] In the mice at week 16, the body weight of the control group was 39.19±0.97 g, the body weight of the COPD model group was 28.36±1.06 g, the body weight of the low-dose syringin group was 31.87±1.30 g, and the body weight of the high-dose syringin group was 34.54±1.35 g.

[0027] From the above results, it can be seen that before modeling, the body weights of all groups were similar, and the difference was not statistically significant (p>0.05), indicating that the grouping was reasonable and the baseline was consistent.

[0028] At the end of modeling, the body weight of the mice in the control group increased significantly, the body weight of the mice in the model group and each treatment group of syringin was significantly lower than that of the control group, which simulated the clinical feature of weight loss in COPD patients.

[0029] After the end of treatment, the body weight of the mice in the control group increased steadily, which reflected healthy growth. The body weight of the mice in the model group increased only slightly, which was significantly lower than that of the control group, indicating that the disease continued to inhibit the increase in body weight. The body weight of the mice in the low-dose syringin group and the high-dose syringin group increased significantly compared with the model group, indicating that syringin treatment can effectively alleviate the weight loss caused by COPD.

[0030] Example 2 Evaluation of whether syringin has the effect of treating chronic obstructive pulmonary disease by FEV0.05 / FVC ratio (1) After the end of treatment, 1.25% tribromoethanol was used as an anesthetic to anesthetize the mice in each group, to ensure that the mice entered a deep anesthetic state, and to avoid respiratory depression caused by excessive anesthesia.

[0031] (2) After the mice were anesthetized, the mice were fixed in a supine position, a surgical lamp was used to illuminate and expose the neck, and the skin and muscle of the neck were incised to expose the trachea.

[0032] (3) The tracheal tube was inserted and connected to the eSpira FM System lung function detection system to measure the forced expiratory volume at 0.05 seconds (FEV0.05) and forced vital capacity (FVC). Taking the normal control group as the reference, the FEV0.05 / FVC ratio was calculated.

[0033] In the results of Figure 2 , the FEV0.05 / FVC ratio of the model group was 0.58±0.04, the FEV0.05 / FVC ratio of the low-dose syringin group was 0.74±0.04, and the FEV0.05 / FVC ratio of the high-dose syringin group was 0.85±0.05.

[0034] From the above results, it can be seen that the FEV0.05 / FVC ratio of the model group decreased significantly compared with the control group, indicating that the COPD mice had significant airflow obstruction, which proved that the modeling of the application was successful. Compared with the model group, the FEV0.05 / FVC ratio of the low-dose syringin group increased by 28%, and the FEV0.05 / FVC ratio of the high-dose syringin group increased by 47%, indicating that after syringin treatment, the airflow obstruction of the COPD mice can be effectively alleviated.

[0035] Example 3 Detection of the effect of syringin on the IL-6 level in the bronchoalveolar lavage fluid (BALF) of COPD mice (1) Keep the mice of each group in which lung function was determined in Example 2 in a state of anesthesia, and inject 0.6 mL of ice-cold normal saline (4°C) from a tracheal catheter to lavage 3 times.

[0036] (2) Collect the mouse bronchoalveolar lavage fluid, centrifuge at 1500 r / min at 4°C for 10 min, and collect the supernatant; (3) Determine the IL-6 level of the bronchoalveolar lavage fluid of the mice in each group by an IL-6 ELISA kit.

[0037] In the results of Example 3, Figure 3 the IL-6 level of the bronchoalveolar lavage fluid of the mice in the control group was 18.68 ± 1.32 pg / mL, the IL-6 level of the bronchoalveolar lavage fluid of the mice in the model group was 90.02 ± 3.11 pg / mL, the IL-6 level of the bronchoalveolar lavage fluid of the mice in the low-dose syringin group was 51.96 ± 2.21 pg / mL, and the IL-6 level of the bronchoalveolar lavage fluid of the mice in the high-dose syringin group was 31.70 ± 2.28 pg / mL.

[0038] From the results of Example 3, Figure 3 it can be seen that the IL-6 level in the BALF of the model group was about 4.8 times that of the control group, indicating that the double stimulation of cigarette smoke + Klebsiella pneumoniae successfully induced typical airway inflammation. The IL-6 level of the low-dose syringin group decreased by 42% compared with the model group, and the IL-6 level of the high-dose syringin group decreased by 65% compared with the model group, indicating that syringin can effectively inhibit the airway inflammation of COPD mice by reducing the level of pro-inflammatory factor IL-6.

[0039] Example 4 Detection of HE staining results of lung tissues of COPD mice after treatment with syringin (1) After the mice in which the bronchoalveolar lavage fluid was collected in Example 3 were fasted and deprived of water for 12 h, they were sacrificed by cervical dislocation, fixed, and the thoracic cavity was opened, and the lung tissue was completely taken out and immediately fixed in a 4% paraformaldehyde solution for 24 h.

[0040] (2) The tissue was dehydrated, paraffin-embedded, sectioned, HE stained, differentiated and re-stained, dehydrated and transparent, and mounted with neutral balsam according to the conventional steps of HE staining.

[0041] (3) Observation was carried out under a microscope, and photographs were taken and recorded.

[0042] From the results of Example 4, Figure 4 it can be seen that the alveolar cavity size of the control group was uniform, the structure was complete, the alveolar septum was thin and continuous, the lung tissue arrangement was dense and regular, and the air cavity and septum ratio was normal, which was the typical morphology of the lung tissue of healthy mice.

[0043] The alveolar cavity of the model group was significantly enlarged, irregular, partially fused to form large cystic cavities, the alveolar septum was obviously thinned or broken, and the lung tissue was loose, suggesting emphysematous changes. At the same time, the local tissue sparseness increased, suggesting successful modeling of COPD.

[0044] Compared with the model group, the alveolar cavity enlargement and fusion phenomenon was reduced in the low-dose syringin group, the alveolar structure was relatively more complete, and the interval damage was reduced, but it was still more disordered than the control group. It is shown that low-dose syringin has a certain improvement effect on lung tissue damage, but the effect is relatively limited.

[0045] The alveolar cavity size of the high-dose syringin group was relatively uniform, the alveolar septum was relatively complete, the lung tissue sparseness was improved obviously, and the emphysematous changes were further reduced, indicating that high-dose syringin can significantly reduce the pathological damage of lung tissue caused by COPD and play a repair role.

[0046] Example 5 Detecting whether the combination of syringin and aminophylline can obtain better therapeutic effect for COPD Aminophylline is a drug for COPD, but its therapeutic window is narrow, and it can easily cause arrhythmia, gastrointestinal reactions and other side effects, so its clinical application is limited. Therefore, this embodiment attempts to combine syringin with aminophylline in order to reduce the dosage of aminophylline while achieving better therapeutic effect. In order to better evaluate the effect of the combination of the two, low-dose syringin and aminophylline are combined.

[0047] (1) COPD modeling is the same as in Example 1, and the experimental groups are as follows: Normal control group (Control): exposed to filtered air, intranasal instillation of the same volume of normal saline, and oral gavage of normal saline; COPD model group (CS+KP): cigarette smoke exposure + Klebsiella pneumoniae instillation, and oral gavage of normal saline; Syringin group: oral gavage of 20 mg / kg syringin once a day for 8 weeks of continuous treatment; Aminophylline group: oral gavage of 10 mg / kg aminophylline once a day for 8 weeks of continuous treatment; Combined treatment group: oral gavage of 20 mg / kg syringin and 10 mg / kg aminophylline once a day for 8 weeks of continuous treatment; The treatment method is the same as in Example 1.

[0048] (2) After the treatment, the body weight of each group of mice at 0, 8 and 16 weeks was measured.

[0049] From Figure 5The results can be seen that, at the 0th week, the initial body weight of each group had no difference; at the 8th week, the body weight of the model group and each treatment group was obviously lower than that of the control group, which was in line with the pathological characteristics of body weight decrease after COPD modeling.

[0050] At the 16th week, the body weight of the control group was 39.66±1.09g, the body weight of the COPD model group was 28.54±0.99g, the body weight of the syringin group was 31.72±1.02g, the body weight of the aminophylline group was 30.01±1.10g, and the body weight of the combined treatment group was 36.97±1.17g.

[0051] From the above results, it can be seen that the body weight of the syringin group was increased by 11.4% compared with the model group, the body weight of the aminophylline group was increased by 5.15% compared with the model group, and the body weight of the combined treatment group was increased by 29.54% compared with the model group. It can be seen that the weight increase effect of the combined treatment group is significantly better than that of the syringin group and the aminophylline group, and also significantly better than the simple addition effect of the two.

[0052] (3) The effect of combined treatment on relieving airflow obstruction of COPD mice was evaluated by FEV0.05 / FVC ratio The detection method was the same as that of Example 2, which will not be repeated here.

[0053] The results obtained by detection are shown in Figure 6 .

[0054] In the Figure 6 , the FEV0.05 / FVC of the COPD model group was 0.59±0.05, the FEV0.05 / FVC of the syringin group was 0.73±0.06, the FEV0.05 / FVC of the aminophylline group was 0.68±0.05, and the FEV0.05 / FVC of the combined treatment group was 0.92±0.04. Among them, the FEV0.05 / FVC percentage of the syringin group increased by 23.73% compared with the model group, the FEV0.05 / FVC percentage of the aminophylline group increased by 15.25% compared with the model group, and the FEV0.05 / FVC percentage of the combined treatment group increased by 55.93% compared with the model group, which was significantly higher than the theoretical addition effect of the two, 2.36 times the effect of the syringin group, and 3.67 times the effect of the aminophylline group, showing a synergistic effect when the two drugs were combined.

[0055] (4) The effect of combined treatment on IL-6 level in bronchoalveolar lavage fluid (BALF) of COPD mice was detected The detection method was the same as that of Example 3, which will not be repeated here.

[0056] The results obtained are shown in Figure 7 .

[0057] In Figure 7 the control group, the IL-6 level of the mice in the bronchoalveolar lavage fluid was 18.36 ± 1.43 pg / mL, the IL-6 level of the mice in the COPD model group was 88.10 ± 2.82 pg / mL, the IL-6 level of the mice in the Syringin group was 51.49 ± 2.67 pg / mL, the IL-6 level of the mice in the Aminophylline group was 65.31 ± 2.26 pg / mL, and the IL-6 level of the mice in the combination therapy group was 25.89 ± 2.05 pg / mL.

[0058] Among them, the IL-6 level of the Syringin group was reduced by 41.55% compared with the model group, the IL-6 level of the Aminophylline group was reduced by 25.87% compared with the model group, and the IL-6 level of the combination therapy group was reduced by 70.61% compared with the model group. According to the expected effect Ea+Eb-Ea×Eb (wherein Ea is the IL-6 reduction rate of Syringin single drug 41.55%, and Eb is the IL-6 reduction rate of Aminophylline single drug 25.87%), the expected effect of the combination of Syringin and Aminophylline is calculated to be reduced by 56.67%. It can be seen that the actual reduction effect is significantly higher than the expected effect, which further proves that the combination of Syringin and Aminophylline can produce significant synergistic effect.

[0059] In summary, the above experimental results show that the combination of Syringin and Aminophylline exhibits significant synergistic therapeutic effect in the treatment of chronic obstructive pulmonary disease (COPD) in mice. Compared with single drug intervention, combination therapy can more effectively reduce airway inflammation, reduce the level of key inflammatory factor IL-6 in bronchoalveolar lavage fluid (BALF), and improve the body weight of mice, thereby achieving high efficiency and low toxicity effect, and having good conversion prospect and clinical application potential.

Claims

1. Use of syringin in the preparation of a medicament for treating chronic obstructive pulmonary disease, characterized in that: The medicine contains syringin as an active ingredient and a pharmaceutically acceptable carrier or excipient.

2. The use according to claim 1, characterized in that The molecular formula of the syringin is C 17 H 24 O9, molecular weight is 372.37, CAS number is 118-34-3.

3. The use according to claim 2, characterized in that The drug is used to treat chronic obstructive pulmonary disease by relieving airflow obstruction, inhibiting airway inflammation and / or improving lung tissue pathological damage.

4. The use according to claim 3, characterized in that The drug is prepared to be administered at a dosage of 20 mg / kg / day to 40 mg / kg / day of syringin content.

5. A drug for treating chronic obstructive pulmonary disease, characterized in that: The drug is composed of the active ingredient syringin and a pharmaceutically acceptable carrier or excipient; The drug is prepared to be administered at a dosage of 20 mg / kg / day to 40 mg / kg / day of syringin content.

6. The drug according to claim 5, characterized in that The molecular formula of the syringin is C 17 H 24 O9, molecular weight is 372.37, CAS number is 118-34-3; The drug is used to treat chronic obstructive pulmonary disease by relieving airflow obstruction, inhibiting airway inflammation and / or improving lung tissue pathological damage.

7. Use of a composition in the preparation of a combined medicament for treating chronic obstructive pulmonary disease, characterized in that: The composition consists of syringin and aminophylline; In the composition, the mass ratio of syringin to aminophylline is 2:

1.

8. The use according to claim 7, characterized in that The drug was prepared and administered at a dosage of 20 mg / kg / day of syringin and 10 mg / kg / day of aminophylline.

9. A combined therapeutic pharmaceutical composition for treating chronic obstructive pulmonary disease, characterized in that: The combined therapeutic pharmaceutical composition consists of syringin and aminophylline; In the combined therapeutic pharmaceutical composition, the mass ratio of syringin to aminophylline is 2:

1.

10. The combined therapeutic pharmaceutical composition according to claim 9, characterized in that The combined therapeutic pharmaceutical composition is prepared to be administered at a dosage of 20 mg / kg / day of syringin and 10 mg / kg / day of aminophylline.

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