CN101057840B - Application of Vam3 in preparation of medicine for treating chronic obstructive pulmonary disease - Google Patents

Abstract

The invention discloses a new application of Vam3 (Amurensin H) represented by formula (I) in the preparation of medicaments for preventing and/or treating chronic obstructive pulmonary disease.

Description

Application of Vam3 in preparation of medicine for treating chronic obstructive pulmonary disease

technical field

The present invention relates to the new medical application of Vam3 (Amurensin H) shown in formula (I).

Background technique

Chronic obstructive pulmonary disease (chronic obstructive pulmonary disease, COPD) is a common chronic disease worldwide, currently ranking fourth in the global cause of death, according to the World Health Organization forecast, by 2020, COPD will be in the global disease incidence Ranked fifth and third among causes of death.

COPD is a disease characterized by not fully reversible airflow limitation associated with an abnormal inflammatory response of the lungs to noxious gases or noxious particles, with chronic inflammation throughout the airways, lung parenchyma, and pulmonary vessels. The cells involved in COPD include inflammatory cells such as neutrophils, T lymphocytes, and macrophages. When the cells are activated, they release various inflammatory mediators, such as leukotriene B ₄ (LTB ₄ ), interleukin 8 ( IL-8), RANTES, Eotaxin, tumor necrosis factor α (TNFα), matrix metalloproteinase 9 (MMP-9), etc., participate in lung parenchymal destruction, pulmonary vascular and other inflammatory reactions. The pathological changes are characterized by infiltration of inflammatory cells (such as macrophages) in the alveolar space, focal inflammatory cell infiltration around the small bronchus and bronchioles, expansion and destruction of the alveolar space at the edge of the lung tissue, widening of the alveolar wall, swelling of the epithelial cells, Rounded and partly peeled off. COPD is closely related to chronic bronchitis and emphysema, but differs in definition, bronchial asthma is related to atopic allergy, and its airflow limitation is reversible (incomplete airflow limitation occurs when combined with chronic bronchitis reversible). At present, there is no drug that can curb the progressive development of COPD, and the research progress of its drug treatment is quite slow. At present, drugs for treating asthma are often used in clinical practice to treat COPD. Because the pathogenesis of COPD is different from that of asthma, it is difficult to obtain satisfactory curative effect.

Vam3 (Amurensin H), which was isolated for the first time by the applicant from the ethanol extract of Vitis vinifera root, can now be chemically synthesized. No. 03134647.2, Publication No. CN1600304). The anti-COPD effect of Vam3 has not been reported yet.

Contents of the invention

Long-term smoking is the most common pathogenic factor of human COPD. Our laboratory uses cigarettes to induce a simulated mouse COPD model, and intends to study the effect of Vam3 on cigarette-induced COPD in mice.

The purpose of the present invention is to provide the application of synthetic or semi-synthetic ingredients in the preparation of anti-COPD disease medicines. Intragastric administration of Vam3 at doses of 50, 100, and 200 mg/kg inhibited the production of inflammatory cytokines TNFa and IL-8 in BALF of cigarette-induced COPD mice; Inhibitory effect; inhibits the recruitment of inflammatory cells in BALF, mainly neutrophils and macrophages; reverses the pathological changes in the lungs of mice.

Detailed ways

Methods: SPF grade male BALB/c mice, 18-20g, were randomly divided into normal control group, model control group, positive control group (terbutaline, 10mg/kg, intragastric administration), drug treatment group (Vam350 , 100, 200mg/kg, intragastric administration), 10 rats in each group. The mice were placed in a 4L container and smoked (Jinjian brand cigarettes, tar content less than 16mg) every day, five times a week, for four consecutive weeks, administered orally 1 hour before the daily smoke. One hour after the last smoke, the mice were anesthetized with phenobarbital sodium, perfused with 0.6 mL of pre-cooled PBS buffer (pH 7.0), rinsed three times, and sucked out the bronchoalveolar lavage fluid (BALF). After centrifugation at 250×g for 10 min, the supernatant was used to analyze the contents of TNFα, IL-8, RANTES, and Eotaxin. The cells were resuspended in PBS buffer, counted, and stained with Wright-Giemsa after cell smears to analyze the changes in the total number of inflammatory cells, neutrophils, and macrophages in the BALF.

Example 1 Vam3 inhibits the generation of inflammatory mediators in COPD mouse bronchoalveolar lavage fluid (BALF)

(1) Inhibitory effect of Vam3 on TNFα production in BALF of COPD mice

The proinflammatory cytokine TNFα is an initiating factor in the pathogenesis of COPD. The level of TNFα in COPD patients is higher than that of normal people, and the cultured bronchial epithelial cells can secrete TNFα in contact with cigarette smoke. TNFα can promote neutrophil degranulation, induce mucosal cell proliferation and hypersecretion, increase the production of IL-8 in epithelial cells, increase the production of matrix metalloproteinases in macrophages, and promote airway hyperresponsiveness. The purpose of this experiment is to investigate the effect of Vam3 on the production of TNFα in the BALF of COPD mice. The results are shown in Table 1.

Table 1 Effect of Vam3 on TNFa production in BALF of COPD mice

Note: In brackets is the inhibition percentage (IR%); ^* P<0.05, compared with the model control group; ^## P<0.01, compared with the normal control group. n=10.

Results: Vam3 at doses of 50, 100, and 200 mg/kg was administered orally to COPD model mice, which could reduce the production of TNFa in the BALF of mice.

(2) Inhibitory effect of Vam3 on IL-8 production in BALF of COPD mice

Lung epithelial cells, fibroblasts, endothelial cells and alveolar macrophages can express IL-8 under the induction of cigarettes. IL-8 is an important mediator of neutrophil chemotaxis, endothelial cell adhesion and degranulation. IL-8 It can activate neutrophil 5-lipoxygenase, chemoattract CD8 ⁺ T cells, and play an important role in the pathological process of COPD. The purpose of this experiment is to investigate the effect of Vam3 on the production of IL-8 in the BALF of COPD mice. The results are shown in Table 2.

Table 2 Effect of Vam3 on IL-8 in BALF of COPD mice

Note: In brackets is the inhibition percentage (IR%); ^* P<0.05, compared with the model control group; ^## P<0.01, compared with the normal control group. n=10.

Results: Vam3 at doses of 50, 100, and 200 mg/kg was administered orally to COPD model mice, which could reduce the production of IL-8 in the BALF of mice.

(3) Inhibitory effect of Vam3 on RANTES secretion in BALF of COPD mice

The migration of inflammatory cells is regulated by chemokines, and various structural cells and inflammatory cells can secrete these small molecular proteins. During the pathogenesis of COPD, the secretion of RANTES increases, chemoattracting monocytes/macrophages, T lymphocytes, and eosinophils cell. The purpose of this experiment is to investigate the effect of Vam3 on the secretion of RANTES in the BALF of COPD mice. The results are shown in Table 3.

Table 3 Effect of Vam3 on RANTES secretion in BALF of COPD mice

Note: In brackets is the inhibition percentage (IR%); ^* P<0.05, compared with the model control group, ^** P<0.01, compared with the model control group; ^## P<0.01, compared with the normal control group. n=10.

Results: Vam3 at doses of 50, 100, and 200 mg/kg was administered orally to COPD model mice, which could reduce the secretion of RANTES in the BALF of the mice.

(4) Inhibitory effect of Vam3 on Eotaxin secretion in BALF of COPD mice

Cells in the allergic inflammatory zone highly express chemokine receptors, and Eotaxin plays an important role in the recruitment of eosinophils to the inflammatory zone by interacting with its specific receptor. In addition, many literatures also reported that Eotaxin plays a role in the migration of related effector cells, such as alveolar macrophages and basophils. The purpose of this experiment is to investigate the effect of Vam3 on the secretion of Eotaxin in the BALF of COPD mice. The results are shown in Table 4.

Table 4 Effect of Vam3 on Eotaxin secretion in BALF of COPD mice

Note: In brackets is the inhibition percentage (IR%); ^* P<0.05, compared with the model control group; ^## P<0.01, compared with the normal control group. n=10.

Results: Vam3 at doses of 50, 100, and 200 mg/kg orally administered to COPD model mice can reduce the secretion of Eotaxin in the BALF of mice.

Example 2 The inhibitory effect of Vam3 on the recruitment of inflammatory cells in the BALF of COPD mice

(1) The inhibitory effect of Vam3 on the recruitment of total inflammatory cells in the BALF of COPD mice

COPD is characterized by chronic inflammation throughout the airways, lung parenchyma, and pulmonary vessels, manifested by an increase in neutrophils, T lymphocytes, and macrophages in the lungs. Activated inflammatory cells release a series of inflammatory mediators, including LTB ₄ , IL-8, TNFa, and others that can damage lung structures or maintain neutrophil inflammation. The purpose of this experiment is to investigate the effect of Vam3 on the recruitment of total inflammatory cells in the BALF of COPD mice. The results are shown in Table 5.

Table 5 Effect of Vam3 on the recruitment of total inflammatory cells in the BALF of COPD mice

Note: In brackets is the inhibition percentage (IR%); ^* P<0.05, compared with the model control group; ^## P<0.01, compared with the normal control group. n=10.

Results: Vam3 administered to COPD model mice at doses of 50, 100, and 200 mg/kg can reduce the recruitment of total inflammatory cells in the BALF of mice.

(2) Inhibitory effect of Vam3 on neutrophil recruitment in BALF of COPD mice

Neutrophils play an important role in the pathogenesis of COPD. They can release two serine proteases, elastase and cysteinyl protease-3, to induce human emphysema-like pathological changes in animals. Neutrophils are short-lived, and their cyclical recruitment to the airways and across the interstitial lumen is rapid. Pathological studies have shown that the increase in the number of neutrophils in the bronchial tissue of some COPD patients is related to the degree of airflow obstruction. The number of neutrophils in the airways is increased in COPD patients who smoke, especially those with chronic bronchitis. The main factor affecting the recruitment of neutrophils in the lungs of COPD patients is the enhanced chemotactic activity of IL-8. The purpose of this experiment is to investigate the effect of Vam3 on the recruitment of neutrophils in the BALF of COPD mice. The results are shown in Table 6.

Table 6 Effect of Vam3 on neutrophil recruitment in BALF of COPD mice

Note: In brackets is the inhibition percentage (IR%); ^* P<0.05, compared with the model control group, ^** P<0.01, compared with the model control group; ^## P<0.01, compared with the normal control group. n=10.

Results: Vam3 at doses of 50, 100, and 200 mg/kg orally administered to COPD model mice can reduce the recruitment of neutrophils in the BALF of mice.

(3) Inhibitory effect of Vam3 on macrophage recruitment in BALF of COPD mice

The level of macrophages in the lungs of smokers and COPD patients is higher than that of normal people, and they mostly gather in alveoli, bronchioles and small airways. The number of alveolar wall macrophages was positively correlated with the degree of small airway disease in patients with mild to moderate emphysema and COPD. Slow progression of COPD and chronic disease paralleled by a long-term increase in macrophages can be seen at sites of tissue disease and injury. Macrophages may lead to an abnormal increase in the degradation capacity of elastic tissue by releasing matrix metalloproteinases, which induces the occurrence of emphysema. The purpose of this experiment is to investigate the effect of Vam3 on the recruitment of macrophages in the BALF of COPD mice. The results are shown in Table 7.

Table 7 Effect of Vam3 on macrophage recruitment in BALF of COPD mice

Note: In brackets is the inhibition percentage (IR%); ^* P<0.05, compared with the model control group; ^## P<0.01, compared with the normal control group. n=10.

Results: Vam3 at doses of 50, 100, and 200 mg/kg orally administered to COPD model mice can reduce the recruitment of macrophages in the BALF of mice.

Example 3. Reversal effect of Vam3 on lung pathological changes in COPD mice

The pathological observation of the mouse lungs showed that the alveolar wall in the model group was widened, the epithelial cells of the alveolar wall were swollen, rounded, and partly shed, macrophages could be seen in the alveolar cavity, focal inflammatory cell infiltration around the small bronchi and bronchioles, lung tissue The marginal alveolar spaces are dilated. In the Vam3 group, the alveolar wall epithelial cells were swollen, and a small amount of macrophages and granulocytes were infiltrated in the alveolar space, but no lymphocyte infiltration was seen, and no hemorrhage was found in the lung tissue of the mice.

Conclusion: Vam3 administered by intragastric administration at doses of 50, 100, and 200 mg/kg has a certain therapeutic effect on chronic obstructive pulmonary congestion and inflammatory cell infiltration in mice induced by cigarettes.

Claims (1)

1. the chemical compound shown in the formula (I) prevents and/or treats application in the medicine of chronic obstructive pulmonary disease as unique active component in preparation