CN115381843B

CN115381843B - Application of L-fucose in preparing medicine for treating respiratory system diseases

Info

Publication number: CN115381843B
Application number: CN202211330879.7A
Authority: CN
Inventors: 肖威; 付娟娟
Original assignee: West China Hospital of Sichuan University
Current assignee: West China Hospital of Sichuan University
Priority date: 2022-10-28
Filing date: 2022-10-28
Publication date: 2023-01-10
Anticipated expiration: 2042-10-28
Also published as: CN115381843A

Abstract

The invention belongs to the technical field of treatment of respiratory system diseases, and relates to application of L-fucose in preparation of a respiratory system disease treatment drug. The research of the invention finds that: lipopolysaccharide and polyinosinic acid can induce the imbalance of the bacterial flora in the mouse airway through airway intervention, and the imbalance of the bacterial flora in the airway participates in promoting airway inflammation; l-fucose atomization inhalation can improve lipopolysaccharide or polyinosinic acid induced acute lung injury model mouse airway inflammation and flora imbalance. The invention confirms that the imbalance of the flora of the airway participates in promoting the airway inflammation, and lays a foundation for treating the airway inflammation by adjusting the flora; and the L-fucose aerosol inhalation administration can obviously improve acute lung injury, and provides a brand new way for treating respiratory system diseases. L-fucose is widely present on plasma membranes on the surfaces of various cells, is safe and harmless, and the atomizing inhalation device is widely used in clinic at present, so that the atomizing of L-fucose is safe, simple, easy and feasible in clinic.

Description

Application of L-fucose in preparing medicine for treating respiratory system diseases

Technical Field

The invention belongs to the technical field of treatment of respiratory system diseases, and particularly relates to application of L-fucose in preparation of a respiratory system disease treatment drug.

Background

The incidence rate of respiratory diseases is high, and the burden of diseases is heavy. Common respiratory diseases, whether acute (such as acute bronchitis, bacterial pneumonia, viral pneumonia and the like) or chronic (such as chronic obstructive pulmonary disease, asthma, bronchiectasis, pulmonary fibrosis and the like), have airway inflammation and impaired airway structure and function to different degrees, so that the improvement of the airway inflammation and the protection of the airway function are one of important treatment methods for treating the respiratory diseases.

Glucocorticoids are currently widely used in respiratory diseases to suppress airway inflammation including chronic obstructive pulmonary disease, asthma, acute lung injury/acute respiratory distress syndrome due to various causes, etc., but they increase the risk of lung infection and have various side effects such as endocrine dyscrasia, peptic ulcer bleeding, femoral head necrosis, etc. There is an urgent clinical need for new safe and effective therapies to ameliorate airway inflammation. Recent clinical studies find that airway flora imbalance exists in various respiratory diseases such as chronic obstructive pulmonary disease, asthma, bronchiectasis, acute respiratory distress syndrome and the like, and the airway flora imbalance is closely related to airway inflammation and disease prognosis. However, the causal relationship between airway flora imbalance and airway inflammation is still lack of reliable experimental confirmation, and at present, no clear and effective method for improving airway flora imbalance so as to achieve the purposes of relieving airway inflammation and treating diseases is available at home and abroad.

Therefore, the technical scheme of the invention is provided.

Disclosure of Invention

In order to solve the problems in the prior art, the invention provides application of L-fucose in preparing a medicament for treating airway inflammation caused by flora imbalance.

At present, no research report on the treatment of respiratory diseases by improving airway flora exists. In view of the fact that various respiratory diseases can cause acute lung injury, such as bacterial and viral (including new corona virus) infection and acute exacerbation of chronic respiratory diseases (including chronic obstructive lung, asthma, bronchiectasis and the like), the invention takes a mouse acute lung injury model as a research object. Through rigorous experimental design, the invention proves that the airway flora imbalance participates in promoting airway inflammation, and the airway atomization L-fucose can improve the airway flora imbalance and relieve the airway inflammation, and the result shows that the L-fucose can be used as an airway prebiotic to treat respiratory system diseases.

L-fucose, a six carbon sugar, is present in greater amounts in seaweed and gums, and is also found in polysaccharides of certain bacteria. L-fucose is widely present on plasma membranes of various cell surfaces as a constituent part of sugar chains in glycoproteins.

The invention provides application of L-fucose in preparing a medicament for treating airway inflammation caused by flora imbalance.

Further, an application of the L-fucose serving as an airway prebiotic in preparing a medicament for treating airway inflammation caused by flora imbalance is provided.

Further, an application of L-fucose serving as an airway prebiotic in preparation of a medicine for treating acute lung injury caused by flora imbalance is provided.

Furthermore, the medicine is a preparation prepared by taking L-fucose as a raw material and adding medically acceptable auxiliary materials or auxiliary components.

Further, the formulation is a formulation for nebulization.

The atomized adjuvants include: phosphate Buffered Saline (PBS).

The invention has the beneficial effects that:

the invention defines that the imbalance of the flora of the airway participates in promoting the airway inflammation, lays a foundation for the treatment of the airway inflammation by adjusting the flora, can obviously improve the acute lung injury by L-fucose atomization, and provides a brand new way for the treatment of respiratory system diseases. L-fucose exists on plasma membranes of various cell surfaces, is safe and harmless, and an atomization inhalation device is widely used clinically at present, so that the clinical atomization of the L-fucose is safe, simple, easy and feasible. Compared with glucocorticoid aerosol inhalation, L-fucose as a prebiotic inhibits inflammation by improving the imbalance of airway flora, so it is expected that it will not increase the risk of pulmonary infection in clinical use and will not produce hormone-related side effects.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a diagram showing information on reagents, instruments and test mice used in examples.

FIG. 2 is a graph of the effect of Lipopolysaccharide (LPS) or polyinosinic acid (Poly: IC) intervention on the α -diversity of mouse airway flora. Wherein:

FIG. 2A is a graph of the effect of LPS or Poly of IC intervention on the Shannon index (Shannon index) of the mouse airway flora.

FIG. 2B is a graph of the effect of LPS or Poly IC intervention on the number of OTUs (Sobs index) observable in the mouse airway flora.

FIG. 2C is a graph of the effect of LPS or Poly IC intervention on the uniformity index (Evenness index) of the mouse airway flora.

Fig. 3 is a table diagram derived from the information of fig. 2.

Figure 4 is a graph demonstrating the effect of LPS or Poly IC intervention on mouse airway flora beta diversity, where:

FIG. 4A is a graph showing the unweighted UniFrac distance of airway flora relative to control after an IC intervention by principal coordinate analysis (PCoA) (the greater the distance of the interclass sample points on the axes, the greater the difference in beta diversity representing interclass airway flora).

FIG. 4B is a graph comparing the distance of airway flora to a control non-weighted UniFrac after LPS or Poly IC intervention.

Fig. 5 is a table diagram derived from the information of fig. 4.

FIG. 6 is a graph of the effect of LPS or Poly IC intervention on the species composition of mouse airway flora.

Fig. 7 is a table diagram derived from the information of fig. 6.

Figure 8 is a graph of the effect of Antibiotic (ABX) intervention on mouse airway bacterial load.

Fig. 9 is a table diagram derived from the information of fig. 8.

FIG. 10 is a graph of the effect of antibiotic intervention on inflammatory cells in alveolar lavage (BAL) of LPS or Poly: IC model mice, in which:

FIG. 10A is a graph of the effect of antibiotic intervention on the total number of inflammatory cells in the BAL of LPS or Poly: IC model mice.

FIG. 10B is a graph of the effect of antibiotic intervention on the total number of neutrophils in the BAL of LPS or Poly: IC model mice.

FIG. 10C is a graph of the effect of antibiotic intervention on total number of macrophages in BAL of LPS or Poly: IC model mice.

Fig. 11 is a table diagram derived from the information of fig. 10.

FIG. 12 is a graph of the effect of antibiotic intervention on LPS or Poly: IC model mouse BAL inflammatory factors, where:

FIG. 12A is a graph of the effect of antibiotic intervention on LPS or Poly: IC model mouse BAL CXCL 1.

FIG. 12B is a graph of the effect of antibiotic intervention on LPS or Poly: IC model mouse BAL GM-CSF.

FIG. 12C is a graph of the effect of antibiotic intervention on LPS or Poly: IC model mouse BAL IL-1 β.

FIG. 12D is a graph of the effect of antibiotic intervention on LPS or Poly: IC model mouse BAL IL-17A.

FIG. 12E is a graph of the effect of antibiotic intervention on LPS or Poly: IC model mouse BAL IL-6.

Fig. 13 is a table diagram derived from the information of fig. 12.

Figure 14 is a graph of the effect of different doses of L-Fucose (Fucose) nebulization on BAL inflammatory cells in mice model of LPS acute lung injury, wherein:

FIG. 14A is a graph of the effect of different doses of L-fucose nebulization inhalation on the total number of BAL inflammatory cells in mice model of LPS acute lung injury.

FIG. 14B is a graph of the effect of different doses of L-fucose nebulization on the total number of BAL neutrophils and macrophages in mice model of LPS acute lung injury.

Fig. 15 is a table diagram derived from the information of fig. 14.

FIGS. 16 and 17 are graphs showing the effect of different doses of L-fucose aerosol inhalation on BAL inflammatory factor in mice model LPS acute lung injury, wherein;

FIG. 16A is a graph of the effect of different doses of L-fucose nebulization inhalation on IL-1. Beta. In BAL of LPS acute lung injury model mice.

FIG. 16B is a graph of the effect of different doses of L-fucose nebulization on IL-17A in BAL of LPS acute lung injury model mice.

FIG. 17A is a graph of the effect of different doses of L-fucose nebulization inhalation on IL-6 in BAL of LPS acute lung injury model mice.

FIG. 17B is a graph of the effect of different doses of L-fucose nebulization on CXCL1 in BAL of LPS acute lung injury model mice.

FIG. 17C is a graph of the effect of different doses of L-fucose nebulized inhalation on GM-CSF in BAL of LPS acute lung injury model mice.

Fig. 18 is a table diagram derived from the information of fig. 16.

Fig. 19 is a table diagram derived from the information of fig. 17.

FIG. 20 is a graph of the effect of different doses of L-fucose nebulization inhalation on the diversity of airway flora alpha in LPS model mice, wherein:

FIG. 20A is a graph of the effect of different doses of L-fucose nebulization on Shannon's index of airway flora in mice model of LPS acute lung injury.

FIG. 20B is a graph of the effect of different doses of L-fucose nebulization on the index of homogeneity of airway flora in mice model of LPS acute lung injury.

FIG. 20C is a graph of the effect of different doses of L-fucose nebulization on the number of OTUs observable in the airway flora of mice model of LPS acute lung injury.

FIG. 21 is a table diagram derived from the information of FIG. 20.

FIG. 22 is a graph of the effect of different doses of L-fucose nebulization on different bacterial populations in the airways of LPS acute lung injury model mice, in which:

FIG. 22A is a graph of the effect of different doses of L-fucose nebulization on Pseudomonas airway (Pseudomonas) in a model mouse of LPS acute lung injury.

FIG. 22B is a graph of the effect of different doses of L-fucose nebulization on Lactobacillus airway (Lactobacillus) in a model mouse with LPS acute lung injury.

FIG. 22C is a graph of the effect of different doses of L-fucose nebulization on the LPS acute lung injury model mouse Campylobacter (Campyleobacteriales) order.

FIG. 23 is a tabular diagram derived from the information of FIG. 22.

Figure 24 is a graph of the effect of L-fucose aerosol inhalation on BAL inflammatory cells in a Poly IC acute lung injury model mouse, in which:

FIG. 24A is a graph of the effect of L-fucose aerosol inhalation on the total number of BAL inflammatory cells in mice model of Poly IC acute lung injury.

FIG. 24B is a graph of the effect of L-fucose aerosol inhalation on the total number of BAL neutrophils and macrophages in mice, a model of Poly: IC acute lung injury.

FIG. 25 is a tabular diagram derived from the information of FIG. 24.

FIGS. 26 and 27 are graphs showing the effect of L-fucose aerosol inhalation on BAL inflammatory factor in Poly IC acute lung injury model mice.

FIG. 26A is a graph of the effect of L-fucose aerosolization inhalation on IL-1. Beta. In BAL of Poly: IC acute lung injury model mice.

FIG. 26B is a graph of the effect of L-fucose aerosol inhalation on IL-17A in BAL of a Poly: IC acute lung injury model mouse.

FIG. 27A is a graph of the effect of L-fucose aerosol inhalation on IL-6 in BAL of a Poly: IC acute lung injury model mouse.

FIG. 27B is a graph of the effect of L-fucose aerosol inhalation on CXCL1 in BAL of Poly IC acute lung injury model mice.

FIG. 27C is a graph of the effect of L-fucose aerosol inhalation on GM-CSF in BAL of a Poly: IC acute lung injury model mouse.

FIG. 28 is a table diagram derived from the information of FIG. 26.

FIG. 29 is a tabular diagram derived from the information of FIG. 27.

Figure 30 is a graph of the effect of L-fucose nebulization inhalation on Poly IC model mouse airway flora alpha diversity, wherein:

FIG. 30A is a graph of the effect of L-fucose aerosol inhalation on the Shannon index of the airway flora in mice model of Poly IC acute lung injury.

FIG. 30B is a graph of the effect of L-fucose aerosol inhalation on the number of OTUs observable in the airway flora of mice as a model of Poly IC acute lung injury.

FIG. 30C is a graph of the effect of L-fucose aerosol inhalation on the index of homogeneity of airway flora in mice as a model of Poly IC acute lung injury.

FIG. 31 is a table diagram derived from the information of FIG. 30.

FIG. 32 is a graph of the effect of L-fucose aerosol inhalation on the composition of airway flora in mice as a model of Poly IC acute lung injury.

FIG. 32A is a graph of the effect of L-fucose aerosolization inhalation on Poly: IC acute lung injury model mouse airways Actinomycetales.

FIG. 32B is a graph of the effect of L-fucose aerosol inhalation on Poly IC acute lung injury model mouse Lactobacillus airway (Lactobacillus).

FIG. 32C is a graph of the effect of L-fucose aerosol inhalation on Poly IC acute lung injury model mouse airway Clostridiales (Clostridiales).

FIG. 33 is a table diagram derived from the information of FIG. 32.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention.

To facilitate understanding of the embodiments, some information in the table will be described.

PBS = phosphate buffer solution; LPS = Lipopolysaccharide, lipopolysaccharide; poly IC = Polyinosinic: polycytidylic acid, polyinosinic acid; WT = wild type, wild mouse; ABX = Antibiotics, antibiotics; BAL = bronchalviolator lavage, alveolar lavage fluid; * P <0.05, P <0.01, P <0.001, P <0.0001; ns = not significant difference.

The information on the reagents, instruments and test mice used in the examples is shown in FIG. 1.

Example 1 demonstrates the correspondence between airway flora imbalance and airway inflammation.

18 mice were randomly divided into 3 groups (6 per group), one of which was a PBS control group and the other two groups were LPS and Poly: IC dried groups, respectively. All mice were general anesthetized by isoflurane induction, model acute lung injury by airway nebulization of LPS (1. Mu.g/g body weight) or Poly: IC (2. Mu.g/g body weight), control group nebulized PBS.

(1) The data information reflected in fig. 2 (fig. 2A to 2C) is shown in fig. 3.

As can be seen from FIGS. 2 and 3, the Shannon index, the uniformity index and the observable OTU number are all important indexes for reflecting the alpha diversity of the flora, and the chart information shows that the alpha diversity of the mouse airway flora can be obviously increased by the intervention of LPS and Poly: IC.

(2) The data information reflected in fig. 4 (fig. 4A to 4B) is shown in fig. 5.

As can be seen from FIGS. 4 and 5, LPS and Poly: IC intervention both resulted in significant changes in the beta-diversity of the mouse airway flora.

(3) The data information reflected in fig. 6 is shown in fig. 7.

As can be seen from FIGS. 6 and 7, the interference of LPS and Poly: IC can cause the composition of mouse airway flora to be changed significantly, wherein the relative abundance of Pseudomonas is reduced significantly after the interference of LPS and Poly: IC, and the relative abundance of Lactobacillus and Campylobacter is increased significantly after the interference of LPS and Poly: IC.

(4) The data information reflected in fig. 8 is shown in fig. 9.

Specifically, antibiotics (2 weeks of amoxicillin and clavulanate potassium +2 weeks of enrofloxacin) were added to the mice drinking water to knock down the airway flora. Fig. 8 and 9 illustrate that this antibiotic intervention regimen significantly reduced the total bacterial load in the mouse airway, and from the data it can be seen that Antibiotic (ABX) intervention significantly reduced the bacterial load in the mouse by a factor of about 1205.

(5) The data information reflected in fig. 10 (fig. 10A to 10C) is shown in fig. 11.

As can be seen from FIGS. 10 and 11, antibiotic intervention significantly reduced the total number of inflammatory cells, the number of neutrophils, and the number of macrophages in the alveolar lavage fluid of LPS and Poly IC model mice, i.e., confirmed that the airway flora was involved in promoting the aggregation of inflammatory cells in the airways of LPS and Poly IC model mice.

(6) Fig. 13 shows data information reflected in fig. 12 (fig. 12A to 12E).

As can be seen from FIGS. 12 and 13, antibiotic intervention significantly reduced the concentration of inflammatory factors in the alveolar lavage fluid of LPS and Poly IC model mice, i.e., confirmed that the airway flora was involved in promoting the release of inflammatory factors in the airways of LPS and Poly IC model mice.

Namely, example 1 can be concluded as follows:

(1) It is clear that both LPS and Poly IC intervention can induce the imbalance of mouse airway flora.

(2) It is clear that an imbalance of airway flora is involved in promoting airway inflammation.

Example 2

Through the research on the effect of the L-fucose atomized by the air passage in an acute lung injury model.

28 mice were randomly divided into 7 groups (4 mice each), one of which was a control group, and the other six groups were different concentrations of L-fucose dry groups. After general anesthesia of all mice induced by isoflurane, acute lung injury model is caused by airway atomization of lipopolysaccharide (1 mug/g body weight), L-fucose intervention group is atomized with different doses (62.5 mug/mouse, 125 mug/mouse, 250 mug/mouse, 500 mug/mouse, 1000 mug/mouse and 2000 mug/mouse) of L-fucose twice (interval 24 h) by airway, and PBS is atomized in control group.

(1) The data information reflected in fig. 14 (fig. 14A to 14B) is shown in fig. 15.

From fig. 14 and fig. 15, it is demonstrated that L-fucose aerosol inhalation can significantly inhibit the aggregation of airway inflammatory cells in mice model of LPS acute lung injury, and is dose-dependent.

(2) The data information reflected in fig. 16 (fig. 16A to 16B) is shown in fig. 18.

(3) The data information reflected in fig. 17 (fig. 17A to 17C) is shown in fig. 19.

From fig. 16 to fig. 19, it is shown that L-fucose aerosol inhalation can significantly inhibit the release of inflammatory factors in the airways of mice in the LPS acute lung injury model, and is dose-dependent.

(4) The data information reflected in fig. 20 (fig. 20A to 20C) is shown in fig. 21.

As can be seen from fig. 20 and 21, it is demonstrated that L-fucose aerosol inhalation can significantly reduce the shannon index and the uniformity index of the airway flora of mice in the LPS acute lung injury model.

(5) The data information reflected in fig. 22 (fig. 22A to 22C) is shown in fig. 23.

As can be seen from fig. 22 and 23, it is demonstrated that L-fucose aerosol inhalation can significantly increase the relative abundance of pseudomonas (pseudomonas) and reduce the relative abundance of Lactobacillales and campylobacter (campylobacter) in the airway flora.

(6) The data information reflected in fig. 24 (fig. 24A to 24B) is shown in fig. 25.

As can be seen from FIGS. 24 and 25, it is demonstrated that L-fucose aerosol inhalation can significantly inhibit aggregation of airway inflammatory cells in mice model Poly: IC.

(7) Fig. 28 shows data information reflected in fig. 26 (fig. 26A to 26B).

(8) The data information reflected in fig. 27 (fig. 27A to 27C) is shown in fig. 29.

As can be seen from FIGS. 26 to 29, L-fucose aerosol inhalation can significantly inhibit release of inflammatory factors in mouse airways of acute lung injury model Poly (IC).

(9) The data information reflected in fig. 30 (fig. 30A to 30C) is shown in fig. 31.

As can be seen from FIGS. 30 and 31, it is demonstrated that L-fucose aerosol inhalation can significantly reduce Poly, the Shannon index, the uniformity index and the number of observable OTUs of the airway flora of the mouse model of IC acute lung injury.

(10) The data information reflected in fig. 32 (fig. 32A to 32C) is shown in fig. 33.

As can be seen from fig. 32 and fig. 33, L-Fucose (fusose) nebulization inhalation significantly increased the relative abundance of Poly (IC-induced acute lung injury model mouse airway actinomycetes) (actinomycetes) and significantly decreased the relative abundance of Lactobacillales (Lactobacillales) and Clostridiales (Clostridiales).

Namely, example 2 can be concluded as follows:

(1) It is clear that L-Fucose (Fucose) aerosol inhalation can improve LPS-induced acute lung injury model mouse airway inflammation and flora imbalance, and the optimal dose is 500 mug/mouse.

(2) On the basis of the optimal dosage, L-Fucose (Fucose) atomization inhalation is confirmed to improve Poly: IC-induced acute lung injury model mouse airway inflammation and flora imbalance.

The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention, and all the changes or substitutions should be covered within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.

Claims

1.L-fucose in the preparation of a medicament for the treatment of airway inflammation caused by an imbalance of flora, said medicament being an aerosolized formulation.

2.L-fucose, in the preparation of a medicament for the treatment of acute lung injury caused by a dysbacteriosis, is an aerosolized formulation.

3. The use of claim 1 or 2, wherein the medicament is an atomized preparation prepared by adding pharmaceutically acceptable auxiliary materials or auxiliary components into L-fucose serving as a raw material.