KR20200080100A - Method of manufacturing and the use of Cordyceps cicadae mycelia active substance for preventing and/or improving acute lung injury - Google Patents

Abstract

The present invention provides a method for preparing a Cordyceps mycelium active substance for preventing and/or alleviating acute lung injury. The method includes the steps of: (a) culturing Cordyceps mycelia in a plate culture medium at 15-30°C for 1-2 weeks; (b) inoculating the Cordyceps mycelia of the step (a) to a flask and culturing the mycelia at 15-30°C and pH 2-6 for 3-14 days; and (c) inoculating the Cordyceps mycelia of the step (b) to a fermenter tank and culturing the mycelia by agitating the same at 15-30°C and pH 2-6 for 3-21 days to obtain a fermented liquor of the Cordyceps mycelium active substance.

Description

Method of manufacturing and the use of Cordyceps cicadae mycelia active substance for preventing and/or improving acute lung injury}

Cordyceps mycelium active material of the present invention, Cordyceps (C Cordyceps cicadae ) relates to a method for producing a mycelium active material and its use. In particular, the present invention relates to a Cordyceps mycelium active material for preventing and/or ameliorating acute lung injury, a method for preparing the Cordyceps mycelium active material and its use in food or pharmaceuticals.

According to Taiwan's Ministry of Health and Welfare, four of the top 10 causes of death in 2017 are lung inflammation, including cancer of the trachea, bronchi, and lungs: malignant neoplasms (1st), pneumonia (3rd), and chronic lower respiratory disease (7th) It is closely related to

Three of the top 10 causes of death in the United States are related to lung disease: lung cancer and bronchial carcinoma, chronic respiratory disease, influenza and pneumonia. Statistical analysis showed that in the United States, 79 out of 100,000 people suffered from acute lung injury (ALI), 59 out of 100,000 suffered from acute respiratory distress syndrome (ARDS), 43% of all deaths, or 75,000 deaths per year, ALI It was further indicated that /ARDS caused it. It is estimated that in the United States, the incidence of ALI/ARDS will double at least in 25 years, as air pollution levels continue to increase and life expectancy increases yearly.

ALI and ARDS (more severe than ALI) are acute inflammatory lung diseases that are clinically frequent, both of which can cause respiratory failure and can be fatal. They are also associated with many respiratory diseases.

ALI-induced risk factors fall into two main groups: direct and indirect risk factors. Direct risk factors are those derived from the lung, such as infectious pneumonia caused by bacteria or viruses, aspiration of gastric acid or foreign bodies, and bruises of the lungs. On the other hand, indirect risk factors are risk factors that do not originate in the lungs, such as sepsis, chronic use of alcohol and drugs, and transfusion of artificial plasma. Bacterial infection is one of the major risk factors, and one example is infection by Gram-negative bacteria, where the skin consists primarily of endotoxins known as lipopolysaccharides (LPS).

Due to the complex mechanism and high mortality of ALI, there is still a lack of clinically effective drugs that can be used to manage mortality of ALI, but general treatment of ALI is mechanical ventilation, beta-2 adrenergic receptor agonist, anticoagulation, Thrombolysis, surfactant therapy and surgery. Therefore, research into effective treatments for ALI is essential for development.

Cordyceps (also known as chong hua, tu chan hua, hu chan, and chan yong cao) is a bud formed by strains at the front of cicada larvae that are parasitized by Clavicipitaceae , fungal spores, and bacterial-carrying bodies that are parasitic by the genus Cordyceps. It is a fruiting body. Cordyceps is mainly used to treat the baby's pathological overnight crying, palpitations and malaria, as well as to relieve breathing and heat from the body and relieve cramps. It is used in herbal tree, classified symptomatology and Zhong hua yao hai. It is described. It has been found through component analysis that natural cordyceps fruiting bodies and bat moth cordyceps ( Ophiocordyceps sinensis ) are constructed in a similar manner.

Recent pharmaceutical studies have found that Cordyceps sinensis has an immune response modulating, antioxidant, anti-inflammatory, neuroprotective and anti-cancer effect and biological activity. However, there have been no studies on the improvement of ALI using Cordyceps mycelium.

The present invention aims to solve the above problem.

The present invention provides a cordyceps mycelium active material and a method of preparing the cordyceps mycelium active material that can be used to prepare a composition effective for the prevention and/or improvement of acute lung injury. Compared with the conventional drugs and treatment methods, the method for producing the fermentation solution of Cordyceps mycelium active material of the present invention is safer and simpler, and the prepared cordyceps mycelium active material is more natural, safe and effective in improving ALI.

According to one embodiment of the present invention, there is provided a method of manufacturing a cordyceps mycelium active material for improving ALI. This method involves the following steps:

(A) culturing Cordyceps mycelium in a plate medium of 15 to 30 ℃ for 1 to 2 weeks;

(b) inoculating the cordyceps mycelium of step (a) into a flask and incubating the mycelium at 15 to 30°C and pH 2 to 6 for 3 to 14 days; And

(c) inoculating the Cordyceps mycelium of step (b) into a fermenter tank and incubating the mycelium by stirring at 15 to 30°C and pH 2 to 6 for 3 to 21 days to obtain Cordyceps mycelium fermentation broth containing the Cordyceps mycelium active material step.

In one embodiment, the method for preparing the Cordyceps mycelium active material further comprises the step of (d): Cordyceps mycelium powder containing Cordyceps mycelium active material by freeze-drying the Cordyceps mycelium fermentation broth and lyophilizing the freeze-dried product. Steps to get.

In one embodiment, the gas is further supplied to the fermenter tank of step (c), the gas comprising air, oxygen, carbon dioxide, helium or a combination thereof; The pressure of the fermenter tank is 0.5 to 1.0 kg/cm ² , and the gas flow rate is 0.01 to 1.5 VVM.

According to another embodiment of the present invention, a cordyceps mycelium active material prepared using the above method is provided.

In another embodiment of the present invention, there is provided a composition for preventing and/or improving acute lung injury comprising the Cordyceps mycelium active material and a pharmaceutically acceptable carrier, excipient, diluent or adjuvant.

In another embodiment of the invention, there is provided the use of the Cordyceps mycelium active material used to prepare a composition for preventing and/or improving acute lung injury.

In one embodiment, improving acute lung injury, as described above, includes alleviating the pathological symptoms of lung inflammation.

In one embodiment, alleviating the pathological symptoms of lung inflammation, as described above, includes restoring the integrity of the damaged or fused alveoli.

In one embodiment, improving acute lung injury, as described above, includes reducing the protein leakage response.

In one embodiment, improving acute lung injury, as described above, includes reducing the level of infiltration in inflammatory cells.

In one embodiment, the inflammatory cells include white blood cells, phagocytes and/or neutrophils.

To further illustrate the above and other features of the present invention, various exemplary embodiments will be described with reference to the accompanying drawings.

It is included in the content of the present invention.

FIG. 1 shows that the pathological condition of ALI can be improved by administering lyophilized powder of Cordyceps mycelium by intraperitoneal injection and then inducing acute lung inflammation by administering endotoxin by nasal inhalation.
Figure 2 shows that the ALI-induced damage of the alveolar capillary barrier can be improved by inducing acute lung inflammation by administering an intraperitoneal injection of lyophilized powder of Cordyceps mycelium and then administering endotoxin by nasal inhalation.
Figure 3 (A) leukocytes, (B) phagocytes and (C) ALI-induced infiltration in neutrophils induces acute lung inflammation by administering an endotoxin by nasal inhalation after intraperitoneal injection of a freeze-dried powder of Cordyceps mycelium It can be improved by showing.
FIG. 4 shows that the pathological condition of ALI can be improved by orally administering a freeze-dried powder of Cordyceps mycelium and then inducing acute lung inflammation by administering endotoxin by nasal inhalation.
FIG. 5 shows that ALI-induced damage to the alveolar capillary barrier can be improved by inducing acute lung inflammation by orally administering lyophilized powder of Cordyceps mycelium and then administering endotoxin by nasal inhalation.
Figure 6 (A) leukocytes, (B) phagocytes and (C) ALI-induced infiltration in neutrophils can be improved by inducing acute lung inflammation by orally administering a freeze-dried powder of Cordyceps mycelium and then administering endotoxin by nasal inhalation It can be shown.

Example 1: Culture of Cordyceps mycelium

The Cordyceps mycelium of the present invention is obtained through the following steps: Natural Taiwan Cordyceps strains are collected, and the mycelium is separated from the fruiting body and the mycelium is stored in a plate medium by passage culture. The genetic sequence of the strain was confirmed as Cordyceps by the Taiwan Food Industry Research and Development Organization. This strain of the Cordyceps sinensis strain is currently publicly deposited as a BCRC number: MU30106 to the Raw Material Collection and Research Center (BCRC) of the Taiwan Food Industry Research and Development Organization, but the Cordyceps mycelium active material of the present invention is limited to materials prepared from these strains Does not work.

(1) Plate culture: Cordyceps mycelium was inoculated into a plate medium and cultured at 15 to 30°C for 1 to 2 weeks (7 days at 25°C in this example). The components of the medium may include potato dextrose agar (PDA), a carbon source and a nitrogen source, and are not particularly limited.

(2) Incubation in flask: Cordyceps mycelium was scraped from the plate of step (1) and inoculated into the flask. The culture was grown from 15 to 30°C in a shake incubator for 3 to 14 days at pH 2 to 6 to 110 to 130 rpm (120 rpm in this example, 25°C and pH 5 for 7 days). The components of the shake culture are shown in Table 1:

Components of culture medium ingredient Amount used in this example
(weight%) Desired amount range (% by weight) Mixed carbon and nitrogen raw materials One 0.01 to 5 hydrocarbon 2.5 0.01 to 10 Yeast or malt extract 0.8 0.001 to 2 Animal and plant proteins and their hydrolysates 0.5 0.01 to 2 Inorganic salt 0.05 0.0001 to 0.05

Among the above components, the mixed carbon and nitrogen raw material may be grain (eg, wheat flour) or legume (eg, soybean powder, mung bean powder, glycine max powder or cinnamon powder); The carbohydrate can be glucose, fructose, maltose, sucrose, etc.; The inorganic salt may be magnesium sulfate, dipotassium phosphate, potassium dihydrogen phosphate, ferric sulfate, and the like. It should be noted that the components shown in Table 1 are only examples, and the components thereof are not specifically limited and can be adjusted according to actual needs or used with commercially available culture media.

(3) Incubation in the fermenter tank: The culture in the flask of step (2) is further inoculated in the fermenter tank, temperature of 15 to 30°C, tank pressure of 0.5 to 1.0 kg/cm ² and 2 to 6 It was stirred at 50 to 150 rpm at pH. Meanwhile, the gas was supplied to the tank at a gas flow rate of 0.1 to 1.0 VVM. The resulting culture was cultured for 3 to 21 days to obtain Cordyceps mycelium fermentation broth (in this example, the culture conditions were 25° C., 0.5 kg/cm ² , pH 5, 80 rpm and 1.0 VVM (air) for 14 days. Was). The medium used in the fermenter tank may contain the same ingredients as the culture of the flask of step (2) or other suitable medium (the same ingredients as in step (2) were used in this example). This microorganism of the Cordyceps sinensis contains the active material of Cordyceps mycelium of the present invention. The freeze-dried powder of Cordyceps mycelium can be obtained by further freeze-drying the Cordyceps mycelium fermentation broth. In this example, 100L of Cordyceps sinensis strain was freeze-dried with 3kg of freeze-dried powder.

The Cordyceps mycelium active material can take a variety of forms including cordyceps strain fermentation broth (strain and clarification liquid), freeze-dried powder made from fermentation broth, or freeze-dried powder dissolved in a solvent or other agent. In a preferred embodiment, the solvent for dissolving the lyophilized powder is water, ethanol or a combination thereof. In a preferred embodiment, the ratio of water to ethanol used as a solvent for dissolving the lyophilized powder is 1:1. In the following Example 2, a subsequent experiment and analysis was performed using the Cordyceps sinensis active material in the form of a freeze-dried powder prepared from fermentation broth.

Example 2: Analysis of the use of Cordyceps mycelium lyophilized powder to improve ALI

One of the major risk factors for ALI is gram-negative bacterial infection, whose shell consists mainly of endotoxins, also known as lipid polysaccharides (LPS). Of the various pathogens that cause ALI, LPS is widely regarded as the most effective inducer of acute lung inflammation and this induction model is most similar to clinically occurring acute inflammation. Therefore, it is possible to obtain an effect of improving ALI by suppressing immune regulation functions and mechanisms as well as LPS-induced lung inflammation. There have been experiments using animal models of endotoxin-induced ALI. Yunhe Fu et al. (2017), Protective effect of TM6 on LPS-induced acute lung injury in mice, SCIENTIFIC REPORTS , 7:572. A mouse model of acute lung inflammation was established by LPS as described in the publication above. In order to induce acute lung injury, 50 μg/20 μL of LPS was administered intranasally to induce acute lung injury. Pulmonary lesions, total bronchial alveolar lavage fluid cell counts, protein concentrations, and various cytokine changes were analyzed to evaluate the improvements made to ALI by synthetic peptides such as decoy peptides derived from cell permeable TIR domains. In this experiment, an acute lung inflammation mouse model was also established using LPS. Thereafter, analysis of changes in lung lesions, total bronchial alveolar lavage fluid cell counts, protein concentrations, and various cytokines was performed to evaluate the improvement of Cordyceps mycelium active material in ALI.

BALB/c mice were used for this animal experiment and divided into two main groups according to the administration route: intraperitoneal administration and oral administration. The two main groups are four groups: control group without substance (control group), group with LPS only (LPS group), group with both LPS and Cordyceps mycelium active substances (CC mycelium group) and LPS and dexamethasone Further divided into positive control group (DEX group) to which all were administered, a total of 8 groups shown in Table 2 below were made. There were 6 mice in each group, a total of 48 mice.

Group description Route of administration group Substance administered Intraperitoneal Contrast group N/A LPS LPS CC mycelium LPS + CC mycelium 0.25 mg/g mouse body weight (6.25 mg CC mycelium for 25 g body weight mice) DEX LPS + DEX 5 μg/g mouse body weight (125 μg DEX for 25 g body weight mice) oral- Contrast group N/A LPS LPS CC mycelium LPS + CC mycelium 0.25 mg/g mouse body weight (6.25 mg of CC mycelium for 25 g body weight mice) (once a day, 3 times in total) DEX LPS + DEX 5 μg/g mouse body weight (125 μg DEX for 25 g body weight mice) (once a day, 3 times in total)

In the intraperitoneal administration group, 20 μl of LPS was administered intranasally, and the concentration of LPS was 50 μg per mouse. Mice were sacrificed 24 hours after administration and samples were collected and analyzed as follows. In the CC mycelium group, the lyophilized powder of Cordyceps mycelium was dissolved in a water/ethanol solvent at a ratio of 1:1. The resulting mixture was homogenized using sonication and used as a sample to be administered when it was confirmed that no precipitate was formed. The Cordyceps sinensis samples were administered intraperitoneally to the mice, and endotoxin (LPS) was administered intranasally 30 minutes after the Cordyceps sinus samples administration using the same administration and sacrificial methods as described above. In the DEX group, DEX was dissolved in physiological saline and 50 μL of the resulting mixture was administered intraperitoneally to mice. Endotoxin (LPS) was administered intranasally 30 minutes after DEX sample administration using the same administration and sacrificial methods as described above.

In the oral administration group, 20 μl of endotoxin (LPS) was administered intranasally, and the concentration of LPS was 50 μg per mouse. Mice were sacrificed 24 hours after administration and samples were collected and analyzed as follows. In the CC mycelium group, the lyophilized powder of Cordyceps mycelium was dissolved in a water/ethanol solvent at a ratio of 1:1. The resulting mixture was homogenized using sonication and used as a sample to be administered when it was confirmed that no precipitate was formed. 200 μL of Cordyceps sinensis samples were administered to the mice once a day through the feed tube for 3 days. Endotoxin (LPS) was administered intranasally 3 days later using the same administration and sacrificial method as described above. In the DEX group, DEX was dissolved in physiological saline, and the resulting mixture was administered to mice through a feed tube once a day. Endotoxin (LPS) was administered intranasally 3 days later using the same administration and sacrificial method as described above.

Histopathological observation of lung tissue: Right lungs of mice not treated with alveolar lavage were extracted and immediately fixed in formalin. The immobilized product was made into paraffin sections and stained with conventional hematoxylin and eosin dyes (H & E dyes). Histopathological changes were observed using an optical microscope.

Bronchoalveolar lavage (BAL): Lung lavage was performed on the left lung of the mouse with 1 mL of PBS using endotracheal intubation to wash the contents of the alveoli and obtain BAL fluid (BALF). BALF was centrifuged and the precipitated cells were measured using different blood cell numbers. Protein concentrations in the supernatant were determined using Bradford protein analysis.

Measurement of leukocyte count and type using flow cytometry: flow cytometry is a highly sensitive quantification method that only causes small personal errors to measure leukocyte differences in blood and BALF samples based on antigens with specificities present on the surface of the white blood cells to be. Measurement targets include: (1) Cell number of BALF; (2) white blood cell (CD45) number; (3) phagocyte (CD45+/CD11b) number; (4) neutrophil (CD45+/Ly6G+) number; And (5) macrophage [(CD45+/CD11b+)-(CD45+/Ly6G+)] number.

Statistical Method: Data are expressed as means±standard deviation (SD) and analyzed using one-way ANOVA. Group differences are analyzed using the LSD post-hoc test. Results with p values less than 0.05 are considered statistically significant.

As shown in FIG. 1, intraperitoneal injection of a freeze-dried powder of Cordyceps mycelium has been found to be effective in improving inflammatory lung disease. No significant change was observed in the control group, where most of the alveolar structures were intact. In the LPS group, pathological changes in the lungs were clearly observed and the alveolar structures were damaged and fused. On the other hand, lung disease in CC mycelium and DEX group mice was clearly improved.

After intraperitoneal injection, protein leakage responses caused by endotoxin-induced lung inflammation were analyzed. It has been found that the lyophilized powder of Cordyceps mycelium is effective in reducing the protein leakage response as shown in Figure 2 and Table 3 below. # P <0.05 compared to control group; *P<0.05 compared to LPS group.

Protein leakage reaction after intraperitoneal injection group Protein leak (mg/mL) Contrast group 2.87±1.02 LPS 9.93±1.52# CC mycelium 5.39±1.27* DEX 3.83±1.72*

(n=6)

# Statistical significance compared to control group (p <0.05)

* Statistical significance compared with LPS group (p <0.05)

After intraperitoneal injection, infiltration responses of (A) leukocytes (CD45) (B) phagocytes (CD45+/CD11b+) and (C) neutrophils (CD45+/Ly6G+) caused by endotoxin-induced lung inflammation were analyzed. It has been found that the freeze-dried powder of Cordyceps mycelium is effective in reducing the infiltrating response of leukocytes, phagocytes and neutrophils as shown in Figures 3 (A to C) and Tables 4-6 below. # P <0.05 compared to control group; *P<0.05 compared to LPS group.

Leukocyte infiltration reaction after intraperitoneal injection group Leukocyte infiltration (10 ⁵ ) Contrast group 0.14±0.06 LPS 3.29±1.95# CC mycelium 0.46±0.34* DEX 0.56±0.29*

(n=6)

# Statistical significance compared to control group (p <0.05)

* Statistical significance compared with LPS group (p <0.05)

Phagocyte infiltration reaction after intraperitoneal injection group Phagocyte infiltration (10 ⁵ ) Contrast group 0.07±0.03 LPS 1.62±0.67# CC mycelium 0.25±0.14* DEX 0.27±0.26*

(n=6)

# Statistical significance compared to control group (p <0.05)

* Statistical significance compared with LPS group (p <0.05)

Neutrophil infiltration reaction after intraperitoneal injection group Neutrophil infiltration (10 ⁵ ) Contrast group 0.04±0.02 LPS 1.12±0.80# CC mycelium 0.15±0.10* DEX 0.16±0.16*

(n=6)

# Statistical significance compared to control group (p <0.05)

* Statistical significance compared with LPS group (p <0.05)

As shown in Fig. 4, it has been found that oral administration of the freeze-dried powder of Cordyceps mycelium active material is effective in improving inflammatory lung disease. No significant change was observed in the control group, where most of the alveolar structures were intact. In the LPS group, pathological changes in the lungs were clearly observed and the alveolar structures were damaged and fused. On the other hand, lung disease in CC mycelium and DEX group mice was clearly improved.

After oral administration, protein leakage responses caused by endotoxin-induced lung inflammation were analyzed. It has been found that the lyophilized powder of Cordyceps mycelium active material is effective in reducing the protein leakage response as shown in Figure 5 and Table 7 below. # P <0.05 compared to control group; *P<0.05 compared to LPS group.

Protein leakage response after oral administration group Protein leak (mg/mL) Contrast group 2.60±0.24 LPS 6.65±1.18# CC mycelium 2.48±0.35* DEX 3.11±0.64*

(n=6)

# Statistical significance compared to control group (p <0.05)

* Statistical significance compared with LPS group (p <0.05)

After oral administration, infiltration responses of (A) leukocytes (CD45) (B) phagocytes (CD45+/CD11b+) and (C) neutrophils (CD45+/Ly6G+) caused by endotoxin-induced lung inflammation were analyzed. It has been found that the freeze-dried powder of Cordyceps mycelium is effective in reducing the infiltrating response of leukocytes, phagocytes and neutrophils, as shown in Figure 6 (A-C) and Tables 8-10 below. # P <0.05 compared to control group; *P<0.05 compared to LPS group.

Leukocyte infiltration reaction after oral administration group Leukocyte infiltration (10 ⁵ ) Contrast group 0.38±0.22 LPS 3.17±0.82# CC mycelium 0.70±0.46* DEX 0.88±0.25*

(n=6)

# Statistical significance compared to control group (p <0.05)

* Statistical significance compared with LPS group (p <0.05)

Phagocytic infiltration reaction after oral administration group Phagocyte infiltration (10 ⁵ ) Contrast group 0.06±0.04 LPS 0.62±0.35# CC mycelium 0.11±0.10* DEX 0.08±0.05*

(n=6)

# Statistical significance compared to control group (p <0.05)

* Statistical significance compared with LPS group (p <0.05)

Neutrophil infiltration reaction after oral administration group Neutrophil infiltration (10 ⁵ ) Contrast group 0.02±0.01 LPS 0.29±0.09# CC mycelium 0.04±0.02* DEX 0.04±0.02*

(n=6)

# Statistical significance compared to control group (p <0.05)

* Statistical significance compared with LPS group (p <0.05)

The cordyceps mycelium active material of the present invention was proved to be effective in improving ALI through the above experiment.

Example 3: Composition preparation

The present invention provides a pharmaceutical composition and a composition comprising Cordyceps mycelium active material to be prepared as a health food.

The composition further comprises additives. In a preferred embodiment, the additives may be excipients, preservatives, diluents, fillers, absorption accelerators, sweeteners, lubricants, thickeners or combinations thereof. The excipient may be selected from sodium citrate, calcium carbonate, calcium phosphate, sucrose, or combinations thereof. Preservatives, such as benzyl alcohol, parabens, silicone, or combinations thereof, can extend the shelf life of pharmaceutical compositions. The diluent can be selected from water, ethanol, propylene glycol, glycerol or combinations thereof. The filler can be selected from lactose, high molecular weight polyethylene glycol or combinations thereof. Absorption accelerators can be selected from dimethyl sulfoxide (DMSO), laurocaram, propylene glycol, glycerol, polyethylene glycol, or combinations thereof. The sweetener can be selected from acesulfame K, aspartame, saccharin, sucralose, neotime or combinations thereof. The lubricant can be selected from magnesium stearate or gum arabic. The thickener can be cornstarch. In addition to the additives, other may be selected according to actual needs as long as the pharmaceutical effect of the composition is not affected.

Compositions can be developed in a variety of products in the pharmaceutical industry. In a preferred embodiment, the composition is a drug, feed, beverage, nutritional supplement, dairy product or health food.

The composition can take a variety of forms to meet patient needs. In a preferred embodiment, the composition may be in powder, tablet, granule, suppository, microcapsule, ampoule/ampoule, liquid spray or suppository form.

The composition of the present invention can be administered to animals or humans. As long as the effect of the composition is not affected, it can be prepared in any dosage form and can be administered via an appropriate route to animals or humans depending on the dosage form.

When the Cordyceps mycelium active material of the present invention is edible, the form of Composition 1 will be an exemplary example of Cordyceps mycelium active material as described below.

Composition 1: Lyophilized powder of Cordyceps mycelium active material (20% by weight) was well mixed with magnesium stearate (8% by weight) used as a lubricant and silicone (7% by weight) used as a preservative. The resulting mixture was dissolved in pure water (65% by weight) and stored at 4°C for later use. The notation "% by weight" means the weight ratio of each component to the total weight of the composition.

When the Cordyceps mycelium active material of the present invention is used medically in liquid form, the form of Composition 2 described below will be an exemplary example of the Cordyceps mycelium active material.

Composition 2: Sucralose (8% by weight), a lyophilized powder of Cordyceps mycelium active material (20% by weight) as a sweetener, Arabian gum (7% by weight) used as a lubricant, and sucrose (10% by weight) used as an excipient ). The resulting mixture was dissolved in pure water (55% by weight) and stored at 4°C for later use. The notation "% by weight" means the weight ratio of each component to the total weight of the composition.

These examples, although disclosed in the above description, are not intended to limit the invention. Those skilled in the art should be able to appropriately change the features described in the present invention with reference to the above teachings and still be able to achieve the effects claimed by the present application. Accordingly, the claims protecting the present invention should be determined by the appended claims.

Claims (11)

(A) culturing Cordyceps mycelium in a plate medium of 15 to 30 ℃ for 1 to 2 weeks;
(b) inoculating the cordyceps mycelium of step (a) into a flask and culturing the mycelium at 15 to 30°C and pH 2 to 6 for 3 to 14 days; And
(c) inoculating the Cordyceps mycelium of step (b) into a fermenter tank and incubating the mycelium by stirring at 15 to 30°C and pH 2 to 6 for 3 to 21 days to obtain Cordyceps mycelium fermentation broth containing the Cordyceps mycelium active material A method for producing a cordyceps mycelium active substance for preventing and/or improving acute lung injury comprising a step. According to claim 1,
The method further comprises the step of (d): freeze-drying the Cordyceps mycelium fermentation broth and crushing the freeze-dried product to obtain Cordyceps mycelium powder containing the Cordyceps mycelium active material. According to claim 1,
The gas is further supplied to the fermenter tank of step (c), the gas comprising air, oxygen, carbon dioxide, helium or a combination thereof; The pressure of the fermenter tank is 0.5 to 1.0 kg/cm ² and the gas flow rate is 0.01 to 1.5 VVM. A cordyceps mycelium active material prepared using the method of claim 1. A composition for preventing and/or improving acute lung injury comprising the Cordyceps mycelium active material of claim 4 and a pharmaceutically acceptable carrier, excipient, diluent or adjuvant. Use of the Cordyceps mycelium active material of claim 4, which is used to prepare a composition for preventing and/or improving acute lung injury. The method of claim 6,
The use of improving the acute lung injury comprises alleviating the pathological symptoms of lung inflammation. The method of claim 7,
Relieving the pathological symptoms of the lung inflammation includes restoring the integrity of the damaged or fused alveoli. The method of claim 6,
The use of improving the acute lung injury comprises reducing the protein leakage response. The method of claim 6,
The use of improving the acute lung injury comprises reducing the level of infiltration in inflammatory cells. The method of claim 10,
Use of the inflammatory cells comprises white blood cells, phagocytes and/or neutrophils.

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