CN114984066A

CN114984066A - Application of phellinus igniarius in preparation of composition for improving sarcopenia

Info

Publication number: CN114984066A
Application number: CN202110225797.5A
Authority: CN
Inventors: 陈劲初; 李宜蓁; 李宗儒; 吕庭宇; 陈彦博
Original assignee: Grape King Bio Ltd
Current assignee: Grape King Bio Ltd
Priority date: 2021-03-01
Filing date: 2021-03-01
Publication date: 2022-09-02
Anticipated expiration: 2041-03-01
Also published as: CN114984066B

Abstract

The present invention provides the use of Phellinus linteus (Phellinus Linteus) for the preparation of a composition for ameliorating sarcopenia, wherein Phellinus linteus may be, for example, the strain deposited under accession number NITE BP-03321. The composition comprises a fermented extract of Phellinus linteus and/or a derivative thereof as an active ingredient. The composition can maintain myotube diameter, muscle mass and muscle endurance, and improve sarcopenia.

Description

Application of phellinus igniarius in preparation of composition for improving sarcopenia

Technical Field

The invention relates to application of phellinus igniarius, in particular to application of phellinus igniarius in preparing a composition for improving sarcopenia.

Background

Sarcopenia is one of the common diseases after the elderly. Sarcopenia is characterized by a continuous decrease in the weight and function of the systemic skeletal muscles, which, if it continues to worsen, not only decreases the activity and quality of life of the patient, but also increases the incidence of other diseases, disabilities, falls, and even death.

Causes of sarcopenia include motor degeneration, imbalances in nutrients, reduced protein synthesis, chronic disease and/or inflammatory response. Currently sarcopenia cannot be controlled by drugs, and therefore needs to be delayed or improved by exercise, control of chronic diseases, inflammatory symptoms and supplementation with specific nutrients.

Phellinus linteus (Phellinus linteus), also called as Morus alba L and Morus songaria Linnaeus, is a medicinal fungus of Phellinus genus of Hymenochaetaceae family. Phellinus linteus is grown on the trunk, especially the trunk of Morus plants. Phellinus linteus has low toxicity but many effects, for example, antioxidation, anti-inflammation, immunity improvement, anticancer, liver protection, anti-dementia, cardiovascular disease prevention, anti-allergy (allergic rhinitis, eczema, rheumatoid arthritis), sleep improvement, analgesia (such as menstrual pain), uric acid inhibition, skin care and the like. However, there are currently few studies on the efficacy of phellinus linteus for improving sarcopenia.

Disclosure of Invention

Accordingly, one embodiment of the present invention provides the use of Phellinus linteus (Phellinus Linteus) for preparing a hypomyotonia-ameliorating composition for maintaining myotube diameter, muscle mass and muscle endurance.

According to the above-described embodiment of the present invention, there is provided a use of Phellinus linteus deposited at National Institute of Technology and Evaluation (NIOD) International Patent Organism Depositary (IPOD) under the accession number NITE BP-03321 at 11/12/2020 in the Japanese National Institute of Technology and Evaluation (NITE) for the preparation of a composition for improving sarcopenia. The composition comprises a fermented extract of Phellinus linteus and/or a derivative thereof as an active ingredient.

According to the above embodiment of the present invention, the fermentation extract may be obtained, for example, by subjecting the first mycelium of Phellinus linteus to a multi-stage cultivation step and an extraction step. First, a first mycelium is subjected to a solid-state culture step using a solid-state medium at 15 ℃ to 30 ℃ for 1 week to 2 weeks to obtain a second mycelium. Then, the second mycelium is subjected to a liquid culturing step at 15 to 30 ℃ for 3 to 14 days using the first culture solution to obtain a third mycelium, wherein the first culture solution has an acid-base value of pH 2 to pH 6. Next, the third mycelium is subjected to a fermentation culture step at 15 to 30 ℃ for 3 to 21 days using a second culture solution having an acid-base value of pH 2 to pH 6 to obtain a fermented product.

According to the above embodiments of the present invention, the fermented extract comprises fermented water extract and/or fermented wine extract, and the derivative is selected from the group consisting of fermented water extract, fermented water extract concentrate, fermented wine extract dry product, fermented wine extract concentrate and any combination thereof.

According to the above embodiment of the present invention, the fermentation water extract is obtained by a water extraction step, and the water extraction step comprises hot water extraction of the fermentation product with water at 100 ℃.

According to the above embodiment of the present invention, the fermented wine extract is obtained by an ethanol extraction step, and the ethanol extraction step may include subjecting the fermented wine extract to ultrasonic vibration treatment with ethanol.

According to the above embodiment of the present invention, the effective dose of the fermented water extract to the animal muscle cells may be, for example, 5. mu.g/mL to 15. mu.g/mL.

According to the above embodiment of the present invention, the effective dose of the fermented wine extract to the animal muscle cells may be, for example, 0.5. mu.g/mL to 1.5. mu.g/mL.

According to the above embodiments of the present invention, when the composition is administered to a mouse, the effective dose of the effective component can be, for example, 400mg/kg. body weight (bw)/day to 600mg/kg.

According to the above embodiments of the present invention, when the composition is administered to a human body, the effective dose of the active ingredient can be, for example, 2300mg/60 kg.bw/day to 2500mg/60 kg.bw/day.

According to the above embodiments of the present invention, the composition may be, for example, a pharmaceutical composition or a food composition, and the composition may include, but is not limited to, a food or medically acceptable carrier, excipient, diluent, adjuvant, preservative, filler and/or additive.

Use of Phellinus linteus of the present invention for the preparation of a hypomyosis-ameliorating composition comprising fermented extract of Phellinus linteus and/or its derivative as an effective ingredient, whereby myotube diameter, muscle mass and muscle endurance can be maintained in a subject.

Drawings

The foregoing and other objects, features, and advantages of the invention will be apparent from the following more particular description of the embodiments of the invention, as illustrated in the accompanying drawings in which:

FIGS. 1A-1F are photomicrographs of tissue staining of mouse skeletal muscle cells cultured in medium with or without fermentation extract and dexamethasone according to one embodiment of the invention.

Detailed Description

In view of the above, the present invention provides a use of Phellinus linteus (Phellinus linteus) for preparing a composition for improving sarcopenia, wherein the composition comprises a fermented extract of Phellinus linteus and/or a derivative thereof as an active ingredient to improve sarcopenia.

The above Phellinus linteus may be deposited, for example, at 11/12/2020 by National Institute of Technology and Evaluation (NITE) International Patent Organism Depositary (IPOD), deposit number NITE BP-03321, wherein Phellinus linteus is deposited in the form of mycelium.

The Phellinus linteus mycelium is subjected to multi-stage culture to obtain Phellinus linteus fermented product. Since Phellinus linteus requires different culture conditions (including nutrients and environmental factors) at different growth or differentiation stages, the culture conditions of Phellinus linteus at each growth stage can be regulated through multi-stage culture to obtain more biomass and/or specific effective components.

In one embodiment, the multi-stage culturing step can comprise a solid state culturing step, a liquid state culturing step, and a fermentation culturing step. In detail, the solid-state culturing step may, for example, culture the above-mentioned mycelium (or first mycelium) of phellinus linteus using a solid-state medium to obtain a second mycelium. The solid medium may contain a carbon source, a nitrogen source, and other nutrients necessary for the growth of Phellinus linteus. In one embodiment, the solid medium may be, for example, potato dextrin medium (PDA). In one embodiment, the solid state culturing step can be, for example, at 15 ℃ to 30 ℃ for 1 to 2 weeks.

The liquid culturing step may be performed on the second mycelium, for example, by using a first culture solution to obtain a third mycelium, wherein the pH of the first culture solution may be, for example, pH 2 to pH 6, and the first culture solution may include 1 wt% to 3 wt% of a comprehensive carbon-nitrogen source (e.g., grains and/or beans), 1 wt% to 4 wt% of a saccharide (e.g., a monosaccharide and/or a disaccharide), 0.1 wt% to 1 wt% of a yeast extract, 0.1 wt% to 1 wt% of a peptone, and 0.01 wt% to 0.05 wt% of an inorganic salt (e.g., a phosphate and/or a sulfate). It should be understood that the composition of the first culture solution can be adjusted according to the requirements of the application. In one embodiment, the first culture solution can be cultured at 15 ℃ to 30 ℃ for 3 days to 14 days at a rotation speed of 110rpm to 130 rpm.

The fermentation culture step may be performed for 3 to 21 days, for example, at 15 to 30 ℃ using a second culture solution, the composition of which may be, for example, the same as the first culture solution, or the composition thereof may be appropriately adjusted according to the use requirement, and the pH of the second culture solution may be, for example, 2 to 6, to obtain a fermentation product.

The fermentation culture step is carried out in a fermentation tank. In one embodiment, a gas is introduced into the fermentation tank during the fermentation step, wherein the gas is selected from the group consisting of air, oxygen, carbon dioxide, helium, and any combination thereof. In one embodiment, the groove pressure may be, for example, 0.5kg/cm ² To 1.0kg/cm ² . In one embodiment, the aeration rateThe rate can be, for example, from 0.01 (volume of gas fed in/volume of fermentation broth/min, VVM) to 1.5 VVM. In other embodiments, the rotation speed of the fermentation culture step is 50rpm to 150 rpm.

Then, the above-mentioned fermented product is subjected to an extraction step to obtain a fermented extract. The extraction step can be carried out using existing extraction methods. In one embodiment, the fermentation product is subjected to a solvent extraction step using a polar solvent, wherein the polar solvent comprises water and/or a lower alcohol (e.g., methanol, ethanol, propanol, isopropanol, etc.). It should be noted that, in consideration of the requirements of subsequent applications, it is preferable to perform the water extraction step or the ethanol extraction step by using water or ethanol, respectively, to obtain the fermented water extract or the fermented wine extract.

In one embodiment, the water extraction step may be performed by hot water extraction with boiling water (e.g., 90 ℃ to 100 ℃). In another embodiment, the ethanol extraction step may optionally be combined with ultrasonic vibration treatment, and performed at room temperature (e.g., 10 ℃ to 40 ℃) by using ultrasonic waves of, for example, 500W to 700W and 30kHz to 50 kHz. The time of the above extraction step is not limited. In one embodiment, the hot water extraction treatment is performed for 20 minutes to 40 minutes. In another embodiment, the ethanol extraction step and the ultrasonic vibration treatment are performed for 40 to 80 minutes to obtain more effective components.

In the above embodiments, the ratio of the polar solvent to the weight of the fermentation product is not particularly limited, and may be, for example, 10 times to 30 times, but is not limited thereto.

In one embodiment, between the multi-stage culturing step and the extracting step, a reprocessing process may be performed, wherein the reprocessing process may include a drying step and/or a concentration step to facilitate the subsequent extracting step.

The above-mentioned drying step may be carried out by a conventional drying method, for example: freeze drying, vacuum drying or spray drying. In one embodiment, the fermentation broth is subjected to a drying step to obtain a fermentation broth.

The concentration step can be carried out by a conventional concentration method such as concentration under reduced pressure, evaporation concentration or membrane concentration.

In one embodiment, the extraction step may also include, but is not limited to, the above-mentioned reprocessing to obtain a derivative of the fermentation extract. The derivative may be selected from the group consisting of a fermented water extract dry product, a fermented water extract concentrate, a fermented wine extract dry product, a fermented wine extract concentrate, and any combination thereof.

The fermented extract and/or the derivative thereof are proved to be used for preparing the composition for improving sarcopenia. The term "ameliorating sarcopenia" means that the loss of muscle mass and muscle endurance is delayed after administration of the above composition to a subject, and specific evaluation criteria include effective maintenance of myotube diameter, muscle mass and muscle endurance.

Animal cell experiments prove that the fermented water extract and/or the fermented wine extract have the effect of maintaining the diameter of myotubes. In this embodiment, the effective dose of the fermented water extract to the animal muscle cells may be, for example, 5 μ g/mL to 15 μ g/mL, and the effective dose of the fermented wine extract to the animal muscle cells may be, for example, 0.5 μ g/mL to 1.5 μ g/mL. The above dosage range is sufficient for the fermented water extract and/or fermented wine extract to have the effect of delaying sarcopenia without causing toxicity to muscle cells.

In addition, the fermented water extract and/or fermented wine extract can maintain muscle mass and muscle endurance after being evaluated by animal experiments.

When used, the fermentation extract and/or the derivative thereof can be added into the composition, and the effective dose depends on the type of the fermentation extract and/or the derivative thereof and/or the application object. In one embodiment, the effective dose of the fermented water extract and/or fermented wine extract is 400mg/kg. body weight (bw)/day to 600mg/kg. In one embodiment, the effective dose of the fermented water extract and/or the fermented wine extract can be, for example, 2300mg/60 kg.bw/day to 2500mg/60 kg.bw/day when the fermented water extract and/or the fermented wine extract is administered to a human body.

When used, the aforementioned composition may be, for example, a food composition or a pharmaceutical composition. In one embodiment, the composition may optionally comprise a food or medically acceptable carrier, excipient, diluent, adjuvant, preservative, filler and/or additive.

The present invention is described in more detail with reference to the following examples, which are provided for illustration purposes only and are not intended to limit the scope of the present invention.

EXAMPLE one preparation of fermentation extract of Phellinus linteus and its derivatives

The first mycelium of Phellinus linteus (Phellinus linteus) of this example was deposited in NITE-IPOD (address: 2-5-8120, Mitsumadzu, Kyoto, Japan, K.K.; zip code: 292) 0818) in Japan on 12.11.2020, with a deposit number of NITE BP-03321. The strain is a mycelium isolated from a wild Phellinus linteus fruiting body of China.

The bacteriological characteristics and culture method of Phellinus linteus (NITE BP-03321) are described in Taiwan application No. TW 109129939, which is incorporated herein by reference.

The mycelium of Phellinus linteus deposited at NITE-IPOD (or called first mycelium) was inoculated on potato dextrin medium (PDA) and cultured at 25 ℃ for 7 days to obtain a second mycelium. Then, a part of the second mycelium of Phellinus linteus was scraped off and inoculated into a first culture solution containing 1 wt% of comprehensive carbon nitrogen source, 1.5 wt% of saccharides, 0.3 wt% of yeast extract, 0.3 wt% of peptone and 0.05 wt% of inorganic salts, and subjected to a culturing step at 25 ℃, pH 5 and rotation speed of 120rpm for 7 days to obtain a third mycelium. The comprehensive carbon-nitrogen source is cereals (wheat flour and/or bran powder) and/or beans (soybean flour, mung bean flour, soybean flour and/or cinnamon powder). The saccharide is monosaccharide (glucose and/or fructose) and/or disaccharide (maltose and/or sucrose). The inorganic salt is phosphate (dipotassium hydrogen phosphate, potassium dihydrogen phosphate) and/or sulfate (magnesium sulfate and/or ferric sulfate). The specific formulations of the comprehensive carbon and nitrogen source, the saccharides and the inorganic salts are well known to those skilled in the art, and can be adjusted arbitrarily according to actual requirements without affecting the fermentation process, and are not further described herein.

Then, the first culture was takenInoculating part of third mycelium of Phellinus Linteus in a fermentation tank containing second culture solution (with the same components as the first culture solution) at 25 deg.C, pH 5, and 0.5kg/cm ² The fermentation was carried out for 14 days at an air pressure of 1.0VVM and an air aeration rate of 80rpm, to obtain a fermented product.

And (4) freeze-drying the fermentation product to obtain the fermentation freeze-dried powder. In this example, 3kg of fermentation lyophilized powder can be obtained from 100L of fermentation product.

And then, carrying out an extraction step on the fermented freeze-dried powder to obtain a fermented extract, wherein the fermented extract comprises a fermented water extract and a fermented wine extract. The fermentation water extract is obtained by extracting with distilled water at 100 deg.C for 30 min, wherein the weight ratio of water to fermentation lyophilized powder is 20. After the fermentation water extract was cooled to room temperature, freeze-drying was performed to obtain sample 1.

The fermented wine extract is obtained by subjecting ethanol to ultrasonic vibration treatment at 25 deg.C for 1 hr, wherein the ultrasonic vibration treatment is performed with ultrasonic wave of 600W and 40kHz by ultrasonic cleaner (manufacturer: Delta ultrasonic wave Co., Taiwan; model: DC600H), and the weight ratio of ethanol to fermented lyophilized powder is 20. Then, centrifugation was performed to separate layers, wherein the supernatant was a fermented wine extract. Then, the fermented wine extract was concentrated under reduced pressure to obtain sample 2.

Sample 1 and sample 2 were dissolved in Dimethyl sulfoxide (DMSO), respectively, to facilitate the later-described arrangement.

Example two evaluation of the efficacy of fermented extracts to maintain myotube diameter, muscle mass and muscle endurance

A sarcopenia profile was established using mouse skeletal muscle cells (C2C12 cell line). The sarcopenia mode may be established using the synthetic corticosteroid dexamethasone (dexamethasone), which inhibits the immune system and thus may act as an anti-inflammatory or anti-allergic agent. However, dexamethasone also has side effects that lead to muscle atrophy (including decreased muscle mass and/or decreased muscle endurance), so this example simulates sarcopenia by inducing muscle cell atrophy with dexamethasone.

First, growth medium [ Dulbecco's modified minimal essential medium (DMEM) containing 10% fetal bovine serum ] was used]At 37 ℃ with 5% CO ₂ Culturing the C2C12 cell line, wherein the initial cell density of the C2C12 cell line is 1x10 ⁵ To 2x10 ⁵ one/mL. When the C2C12 cell line grew to 70% full, differentiation culture was performed for 7 days in differentiation medium (DMEM containing 2% horse serum), during which the differentiation medium was changed every 2 days, to obtain differentiated cells, 90% of which were differentiated into myotubes.

Next, the differentiated cells were divided into a blank control group, a negative control group, an experimental group 1, an experimental group 2, an experimental group 3 and an experimental group 4, wherein the medium of the blank control group was DMEM containing 0.1% DMSO, the medium of the negative control group was DMEM containing 0.1% DMSO and 10. mu.M dexamethasone, the medium of the experimental group 1 was DMEM containing 10. mu.g/mL of sample 1, the medium of the experimental group 2 was DMEM containing 10. mu.M dexamethasone and 10. mu.g/mL of sample 1, the medium of the experimental group 3 was DMEM containing 1. mu.g/mL of sample 2, and the medium of the experimental group 4 was DMEM containing 10. mu.M dexamethasone and 1. mu.g/mL of sample 2. It is noted that, since samples 1 and 2 were dissolved in DMSO before being used to prepare the culture medium, the culture medium of panels 1 to 4 contained DMSO at a concentration of 0.1% or less to avoid toxicity to the cells.

At 37 deg.C, 5% CO ₂ Then, after culturing the differentiated cell lines in the above culture medium for 24 hours, hematoxylin and eosin (H) was added&E) The diameter of myotubes in the differentiated cell lines was observed by light microscopy as shown in FIGS. 1A to 1F.

Fig. 1A to 1F are graphs showing tissue staining of mouse skeletal muscle cells cultured in a medium with or without fermentation extract and/or dexamethasone according to an embodiment of the present invention, wherein fig. 1A, 1B, 1C, 1D, 1E and 1F correspond to a blank control group, a negative control group, an experimental group 1, an experimental group 2, an experimental group 3 and an experimental group 4, respectively. The myotube diameters were smaller in fig. 1B (negative control group) compared to fig. 1A (blank control group), but larger in fig. 1C to 1F (experimental group 1 to experimental group 4) compared to fig. 1B (negative control group).

Myotube diameters were measured using a commercially available software (Image-Pro Plus software) and statistical results of myotube diameters are shown in table 1, where the statistical approach of table 1 is to analyze the percentages using paired sample t-test, and "#" and "-" represent statistically significant differences from the blank and negative control group pitches, respectively (p <0.05, n 60).

TABLE 1

As can be seen from table 1, the diameter of myotubes in the negative control group was significantly reduced compared to the blank control group, indicating that dexamethasone did indeed cause muscle cell atrophy. However, the significantly higher levels in experimental group 2 and experimental group 4 compared to the negative control group indicate that the fermented water extract and/or fermented wine extract can maintain the length of myotube diameter induced by dexamethasone, meaning that the loss of muscle mass can be delayed and/or avoided. In addition, there was no significant difference between the experimental group 1 and the experimental group 3 and the blank control group, which indicates that the fermented water extract or the fermented wine extract did not affect the size of the myotube diameter, in other words, the fermented water extract or the fermented wine extract did not have cytotoxicity to the myotube cells.

EXAMPLE III evaluation of the efficacy of fermented extracts to maintain muscle Mass and muscle endurance

The above sample 1 and sample 2 were mixed by equal weight to obtain sample 3.

This example uses C57BL/6J mice as model organisms. The mice were divided into blank, control and experimental groups and were fed with the corresponding reagents once a day, with the reagent of the experimental group being sample 3 and the reagents of the blank and control groups being water. Specifically, sample 3 was dissolved in an appropriate amount of water to control the amount of feed at 500 mg/kg.bw/time, and the volume of water tube-fed to the blank and control groups was equivalent to the total volume of sample 3 mixed with water.

Hind limbs of mice of the experimental group and the control group were subjected to a plaster fixation treatment for 7 days, respectively, to induce atrophy of the hind limbs of the mice. Next, the plaster was removed and the mice were allowed to move freely in the mouse cage for 7 days. Then, a muscular endurance test was performed, and the mouse was sacrificed to measure the mouse hind limb skeletal muscle. Tube feeding was continued during the gypsum fixation treatment and free-play period described above (total 14 days).

The muscle endurance test described above was performed by placing the mice on an inclined treadmill, wherein the conveyor belt of the treadmill was transported downward at a rate of 18m/min to 20m/min, and the bottom of the treadmill was provided with shock grids. If the mouse is immobilized, the conveyor belt will transport the mouse to the bottom of the treadmill, causing the mouse's tail to be shocked. Generally, the mouse runs up to avoid a shock to the tail. However, if the muscle endurance is insufficient, the mouse may not be transported to the bottom of the treadmill against the speed of the conveyor belt, thereby suffering an electric shock. If the mouse receives more shocks within the same period of time, the muscle endurance of the mouse is worse. The number of shocks received by the mice was analyzed using a commercially available software [ GraphPad Prism (version 8.0) ] with single factor variability analysis (one-way ANOVA) and Dunnett's test post hoc assays (post-hoc), and the results are shown in table 2, where "#" and "#" indicate statistically significant differences from the blank and control groups, respectively (p <0.05, n ═ 6).

TABLE 2

Group of	Number of electric shocks
		Blank group	3.00±2.28
Control group	500.83±257.37#
		Experimental group	25.60±48.87*

As shown in table 2, the number of shocks received by the control mice was significantly increased compared to the blank group, indicating that the plaster fixation method can indeed cause skeletal muscle atrophy and loss of muscle endurance in the mice. However, the number of electric shocks received by the mice in the experimental group was significantly reduced compared to the control group, indicating that the application of the mixture of the fermented water extract and the fermented alcohol extract can effectively maintain the muscle mass and the muscle endurance, thereby delaying and/or avoiding the loss of the muscle endurance of the mice.

Next, the mice were sacrificed and the weight of the hind limb gastrocnemius muscles was measured. The results were analyzed by one-way ANOVA (one-way ANOVA) and Dunnett's test post hoc (post-hoc) using commercially available software [ GraphPad Prism (version 8.0) ] to count the differences between groups. The results are shown in table 3, in which the relative weight is the weight of gastrocnemius muscle divided by the mouse weight to exclude factors of individual differences in body type, and the graphs "#" and "-" indicate statistically significant differences (p <0.05, n ═ 6) from the blank group and the control group, respectively.

TABLE 3

Group of	Relative weight (mg/g)
		Blank group	5.485±0.234
Control group	4.895±0.384#
		Experimental group	5.051±0.182*

As shown in table 3, the relative weight of the gastrocnemius muscle was significantly less in the control group compared to the blank group, indicating that the gypsum fixation treatment may cause a decrease in muscle mass. However, the relative weight of gastrocnemius muscle was significantly greater in the experimental group compared to the control group, indicating that the administration of the mixture of fermented water extract and fermented wine extract was effective in maintaining muscle mass, thereby delaying and/or avoiding loss of muscle mass.

EXAMPLE four estimation of effective dose in humans

This example estimates the effective dose of human body from the effective dose of the above mouse according to the initial estimation method of experiment published by the U.S. food and drug administration in 2005. The method is to divide the effective dose of the mouse per kilogram of body weight by a conversion coefficient of 12.3 to obtain the effective dose of the human body per kilogram of body weight. The effective dose in humans is 2400mg/60 kg.bw/day (calculated as the body weight of an adult human being is 60 kg.bw), estimated from the daily dose of 500mg/kg.bw administered to mice in this example.

As described above, the fermented water extract and the fermented wine extract according to the present invention can effectively maintain the diameter of myotubes, the muscle mass and the muscle endurance, and thus can be used for ameliorating sarcopenia.

It should be noted that although the present invention describes the use of phellinus linteus of the present invention for preparing a composition for ameliorating sarcopenia by exemplifying a specific process, a specific analytical method and/or a specific apparatus, those skilled in the art of the present invention will appreciate that the present invention is not limited thereto, and the use of phellinus linteus of the present invention for preparing a composition for ameliorating sarcopenia may be carried out by using other processes, other analytical methods or other apparatuses without departing from the spirit and scope of the present invention.

While the invention has been described with reference to certain embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

[ biological Material Collection ]

Phellinus linteus (Phellinus linteus) is deposited at 11.12.2020 in Japan independent administration of Technology and Evaluation Technology (NITE) International Patent Organism Depositary (IPOD), Address: 2-5-8120 th room of Gentianjin city, Qianye county, Japan; and (3) post code: 292-; deposit number NITE BP-03321. Phellinus linteus is also deposited in the biological resource center (BCRC) of food industry development research institute of Caucao Farmland, Taiwan, Xinzhu food Lu No. 331, and Taiwan, at 7/18.2019, and has a deposit number of BCRC 930210.

Claims

1. Use of Phellinus linteus (Phellinus linteus) for preparing a composition for improving sarcopenia, wherein Phellinus linteus is deposited at National Institute of Technology and Evaluation, NITE (International Patent organization for the advancement of Technology and Evaluation, NIOD) International Patent Organism Depositary (IPOD) at 11/12.2020, and the Phellinus linteus is deposited under the accession number NITE BP-03321, and the composition comprises a fermentation extract and/or a derivative thereof of Phellinus linteus as an effective ingredient for improving sarcopenia.

2. The use of Phellinus linteus Linteus of claim 1 for preparing a composition for improving sarcopenia, wherein the fermentation extract is obtained by subjecting a first mycelium of Phellinus linteus to a multistage culture step and an extraction step, wherein the multistage culture step comprises:

subjecting the first mycelium to a solid state culture step at 15-30 ℃ for 1-2 weeks using a solid state medium to obtain a second mycelium;

performing a liquid culture step on the second mycelium at 15-30 ℃ for 3-14 days using a first culture solution to obtain a third mycelium, wherein an pH of the first culture solution is from pH 2 to pH 6; and

subjecting the third mycelium to a fermentation culture step at 15-30 ℃ for 3-21 days using a second culture solution to obtain the fermentation product, wherein an pH of the second culture solution is from pH 2 to pH 6.

3. The use of Phellinus linteus as claimed in claim 2, wherein the fermented extract comprises a fermented water extract and/or a fermented wine extract, and the derivative is selected from the group consisting of a fermented water extract dried product, a fermented water extract concentrated product, a fermented wine extract dried product, a fermented wine extract concentrated product and any combination thereof.

4. The use of Phellinus linteus as claimed in claim 3, wherein the fermentation water extract is obtained by a water extraction step, and the water extraction step comprises subjecting a fermentation to a hot water extraction treatment with water at 100 ℃.

5. The use of Phellinus linteus as claimed in claim 3, wherein the fermented wine extract is obtained by an ethanol extraction step, and the ethanol extraction step comprises subjecting a fermented product to an ultrasonic vibration treatment with ethanol.

6. The use of Phellinus linteus extract as claimed in claim 2, wherein an effective dose of the fermented water extract to muscle cells of an animal is 5 μ g/mL to 15 μ g/mL.

7. The use of Phellinus linteus extract as claimed in claim 2, wherein an effective dose of fermented wine extract to muscle cells of an animal is 0.5 μ g/mL to 1.5 μ g/mL.

8. The use of Phellinus linteus as claimed in claim 1, wherein an effective dose of the effective component is 400mg/kg. body weight (bw)/day to 600mg/kg.

9. The use of Phellinus linteus for preparing a composition for improving sarcopenia as claimed in claim 1, wherein an effective dose of the effective component is 2300mg/60 kg.bw/day to 2500mg/60 kg.bw/day when the composition is administered to a human body.

10. The use of Phellinus linteus as claimed in claim 1, wherein the composition is a pharmaceutical composition or a food composition, and the composition further comprises a food or a pharmaceutically acceptable carrier, an excipient, a diluent, an adjuvant, a preservative, a filler and/or an additive.