阅读说明：本技术 一种人工栽培桑黄子实体的提取物及其制备方法和应用 (Extract of artificially cultivated phellinus igniarius sporocarp and preparation method and application thereof ) 是由 张信岳 王建功 李钦 吕玮 谷丽丽 王星晨 武柠子 王建文 于 2019-11-18 设计创作，主要内容包括：本发明公开了一种人工栽培桑黄子实体的提取物及其制备方法和应用,该制备方法包括：选取桑黄菌株桑都1号,采用自然林下仿野生袋料栽培方法,当子实体由黄色变成褐色或深褐色时采收,即得桑黄子实体；将桑黄子实体经过粉碎得到粉体,用乙醇水溶液浸泡,回流提取,将提取液减压浓缩至无乙醇味,真空干燥,得到人工栽培桑黄子实体的提取物。本发明提取物可以改善和缓解高尿酸血症小鼠的肾脏损伤,增加尿酸排泄,且作用温和,副作用少,可以用于制备口服的防治高尿酸血症和痛风的药物和保健品。(The invention discloses an extract of artificially cultivated phellinus igniarius sporocarp, a preparation method and application thereof, wherein the preparation method comprises the following steps: selecting Phellinus Linteus strain Phellinus Linteus number 1, adopting wild-like bag cultivation method under natural forest, and collecting when fruiting body changes from yellow to brown or dark brown to obtain Phellinus Linteus fruiting body; pulverizing Phellinus linteus fruiting body to obtain powder, soaking in ethanol water solution, reflux extracting, concentrating the extractive solution under reduced pressure until ethanol smell disappears, and vacuum drying to obtain artificially cultured Phellinus linteus fruiting body extract. The extract can improve and relieve kidney injury of mice with hyperuricemia, increases uric acid excretion, has mild effect and less side effect, and can be used for preparing oral medicaments and health-care products for preventing and treating the hyperuricemia and the gout.)

1. An extract of artificially cultivated phellinus linteus fruiting body and a preparation method thereof, which is characterized by comprising the following steps:

1) selecting Phellinus Linteus strain Phellinus Linteus 1, having the chemical name of Phellinus Linteus sanghuang, adopting wild-imitating bag cultivation method under natural forest, and collecting when sporophore changes from yellow to brown or dark brown to obtain Phellinus Linteus sporophore;

2) crushing the phellinus igniarius sporocarp obtained in the step 1) to obtain powder, soaking the powder in an ethanol water solution, performing reflux extraction, concentrating an extracting solution under reduced pressure until the ethanol smell does not exist, and performing vacuum drying to obtain an extract of the artificially cultured phellinus igniarius sporocarp.

2. The extract from fruiting body of artificially cultivated Phellinus linteus as claimed in claim 1, wherein in step 1), Phellinus linteus strain Phellinus linteus Du 1 is Phellinus linteus mother strain, and is preserved in China general microbiological culture Collection center with preservation number CGCMCC No. 8854.

3. The extract of artificially cultivated phellinus linteus fruiting body according to claim 1, wherein the volume percentage of the ethanol aqueous solution in step 2) is 60-80%.

4. The extract of artificially cultivated phellinus linteus fruit body according to claim 1, wherein the ratio of the amount of the powder to the amount of the ethanol aqueous solution in step 2) is 20 g: 300mL to 800 mL.

5. The extract of the fruiting body of Phellinus linteus cultured artificially according to claim 1, wherein in step 2), the fruiting body is soaked in aqueous solution of ethanol for 0.2-1 h.

6. The extract of artificially cultivated phellinus linteus fruiting body according to claim 1, wherein the temperature of the vacuum drying in step 2) is 50 to 70 ℃.

7. An extract of artificially cultivated phellinus linteus fruit body prepared by the preparation method according to any one of claims 1 to 6.

8. The use of the extract of the fruiting body of Phellinus linteus as claimed in claim 7 in the preparation of a medicine for preventing and treating gout and hyperuricemia.

9. The use of the extract of the fruiting body of Phellinus linteus as claimed in claim 7 in the preparation of health products for preventing and treating hyperuricemia and gout.

10. The use according to claim 8 or 9, wherein the medicament for the prevention and treatment of gout and hyperuricemia and the health care product for preventing and treating hyperuricemia and gout are orally administered.

Technical Field

The invention relates to the field of extracts of phellinus igniarius sporocarp, in particular to an extract of artificially cultured phellinus igniarius sporocarp, a preparation method thereof and new application thereof in preventing and treating hyperuricemia and gout.

Background

Gout belongs to the category of metabolic diseases, and is a series of inflammatory reactions caused by purine substance metabolism disorder in organisms or reduction of uric acid excretion of kidney, so that urate deposition is caused. When uric acid in serum is increased to a supersaturated state, extracellular fluid is deposited on joints, tissues and organs around the joints, so that a group of gout syndromes (including tophus, uric acid kidney stones, gouty arthritis and gouty nephropathy) are caused. According to statistics, the total incidence of gout is 0.06-2.68 per mill. Gout has become the second disease of diabetes in China, and the onset age of gout is in a tendency of being younger. At present, the gout treatment approaches mainly reduce the incidence of hyperuricemia by inhibiting the excessive production of uric acid and promoting the excretion of uric acid, and inhibit inflammatory cytokines and the like. Although the effect of the chemical medicine for treating gout is quick, the adverse reactions are more, for example, the adverse reactions caused by 114 cases of allopurinol collected in 2015 year 2000-. Other drugs such as febuxostat, benzbromarone, colchicine and the like have toxic and side effects of different degrees, including kidney injury, central nervous system toxicity and the like, and have high occurrence rate. Therefore, the search for new drug administration strategies is urgent.

Phellinus igniarius, also called Morganaceae, Morus alba and Phellinus igniarius, belongs to Basidiomyeotinia, Hymenomyetes, Aphylopolorales, Polyporaeeae and Phellinus, belongs to Basidiomycotina, Hymenomyetes, Polyporaeeae and Phellinus, is a kind of medicinal fungus, is mainly parasitic on trunks, stumps or fallen woods of broad-leaved trees such as mulberry, willow, poplar, oak, hawthorn and the like, and is perennial and has the name of forest gold. Phellinus linteus is mainly distributed in northeast, northwest, southwest and other areas of China, and 7 Phellinus linteus groups exist. While there are 1 Phellinus group in Zhejiang, i.e., Phellinus linteus, which is known as Inonotus sanghuang, and grows in the field only on the trunk of Morus (Morus) plants, almost completely. The wild phellinus igniarius has long growth period and small quantity, so that the wild phellinus igniarius has short supply and demand and high price. Compared with some countries such as European Union, Japanese Korea and the like, China still has a large gap for the development and utilization technology of phellinus igniarius products. The phellinus igniarius sporocarp has various active ingredients, the main medicinal ingredients are flavone, polysaccharide, pyrone and triterpenes, and the phellinus igniarius sporocarp has pharmacological functions of resisting tumors, enhancing the immunity of an organism, resisting oxidation, diminishing inflammation, reducing blood sugar and blood fat, resisting pneumonia and the like, but reports on treatment of gout and hyperuricemia are not seen.

Disclosure of Invention

The invention provides an extract of artificially cultured phellinus igniarius sporocarp, a preparation method and application thereof, and the extract is used for preparing medicines and health care products for preventing and treating hyperuricemia and gout.

The technical scheme of the invention is as follows:

an extract of artificially cultivated phellinus linteus fruiting body and a preparation method thereof, comprising the following steps:

1) selecting Phellinus Linteus strain Phellinus Linteus 1, having the chemical name of Phellinus Linteus sanghuang, adopting wild-imitating bag cultivation method under natural forest, and collecting when sporophore changes from yellow to brown or dark brown to obtain Phellinus Linteus sporophore;

2) crushing the phellinus igniarius sporocarp obtained in the step 1) to obtain powder, soaking the powder in an ethanol water solution, performing reflux extraction, concentrating an extracting solution under reduced pressure until the ethanol smell does not exist, and performing vacuum drying to obtain an extract of the artificially cultured phellinus igniarius sporocarp.

In the step 1), the phellinus igniarius strain sondu 1 is phellinus igniarius mother strain, the strain is preserved in China general microbiological culture collection center (CGCMCC) at 03-07 th in 2014, the preservation number is CGCMCC No.8854, the strain is classified and named as phellinus igniarius sanghuang, and the strain is disclosed in Chinese patent ZL 201410477690. X.

In the step 2), the volume percentage of the ethanol water solution is 60-80%. More preferably, the ethanol aqueous solution is 65% to 75% (most preferably 70%) by volume.

The dosage ratio of the powder to the ethanol water solution is 20 g: 300mL to 800mL, more preferably 20 g: 400mL to 600mL, most preferably 20 g: 500 mL.

Soaking the mixture for 0.2-1 h by using an ethanol water solution, and preferably, soaking the mixture for 0.5h by using the ethanol water solution.

The temperature of the vacuum drying is 50 ℃ to 70 ℃, more preferably 55 ℃ to 65 ℃, and most preferably 60 ℃.

The new application of the extract of the artificially cultivated phellinus igniarius sporocarp comprises the following steps: the extract of the artificially cultured phellinus igniarius sporocarp can be used for preparing medicines for preventing and treating gout and hyperuricemia (namely anti-gout and hyperuricemia medicines) and health care products for preventing and treating hyperuricemia and gout.

The artificially cultured Phellinus linteus fruiting body extract is effective for oral administration, and can be administered orally. Namely, the medicine and the health care product for preventing and treating gout and hyperuricemia are orally taken.

The uric acid reducing effect of artificially cultured Phellinus linteus fruiting body extract is related to xanthine oxidase inhibition.

The recommended human dosage range of the artificially cultivated phellinus igniarius sporocarp extract is 5.5-148.5 mg/kg, and the balance is a pharmaceutically acceptable carrier.

The medicine or health care product is a medicine composition taking artificially cultivated phellinus igniarius sporocarp extract as a main active substance, also comprises auxiliary materials acceptable in the field of pharmacy, and is any medicinal preparation form.

Through research, the following results are found: artificially cultured phellinus igniarius sporophore extract (40-160 mu g/mL) remarkably reduces cell damage caused by sodium urate in vitro and improves cell survival rate. In vivo research shows that the extract has a strong uric acid reducing effect, the serum uric acid level and liver xanthine oxidase activity of hyperuricemia mice induced by potassium oxonate can be obviously reduced by intragastrically administering the phellinus igniarius sporocarp extract with a dose of 150mg/kg, and the phellinus igniarius sporocarp extract has no damage to kidneys; the drug is administered by gavage of 20.0g/kg for mice, no animal dies, no obvious toxic or side effect is seen, and the drug can be used for preparing oral anti-gout and hyperuricemia drugs or health care products, and provides a new direction for improving the phenomenon of large side effect of the existing gout drugs.

Compared with the prior art, the invention has the following advantages:

through researches, the artificially cultivated phellinus igniarius sporocarp extract is found to have the obvious effect of reducing uric acid. In vitro cell experiments show that the phellinus igniarius sporocarp extract has a strong protective effect on cell damage induced by sodium urate; the phellinus igniarius sporocarp extract with the dosage of 150mg/kg after gastric lavage can obviously reduce the serum uric acid and creatinine level of a hyperuricemia mouse induced by potassium oxonate and the xanthine oxidase activity in the liver, and has no influence on the weight of the mouse, thereby showing that the invention can improve and relieve the kidney injury of the hyperuricemia mouse, increase the uric acid excretion, has mild effect and little side effect, and can be used for preparing oral medicaments and health care products for preventing and treating hyperuricemia and gout.

The invention applies the artificially cultivated phellinus igniarius sporocarp extract to the preparation of the anti-gout and hyperuricemia drug for the first time, compared with the clinical first-line xanthine oxidase inhibitor allopurinol, although the activity is weaker than allopurinol, the toxicity is lower than allopurinol (mouse oral administration LD)₅₀700mg/kg), the maximum tolerated dose of the extract is greater than 20.0g/kg, corresponding to 100 times the pharmacodynamically effective dose.

Drawings

FIG. 1 is a graph showing the effect of Phellinus linteus fruiting body extract on the survival rate of sodium urate-induced cells;

FIG. 2 is a graph showing a comparison of the effect of Phellinus linteus fruiting body extract on the body weight of hyperuricemia mice;

FIG. 3 is a graph showing a comparison of the effect of Phellinus linteus fruiting body extract on the serum uric acid level of hyperuricemia mice;

FIG. 4 is a graph showing a comparison of the effect of Phellinus linteus fruiting body extract on serum creatinine level in hyperuricemia mice;

FIG. 5 is a graph showing a comparison of the effect of Phellinus linteus fruiting body extract on xanthine oxidase activity in liver of hyperuricemia mouse;

FIG. 6 is a graph showing the effect of Phellinus linteus fruiting body extract on body weight of normal mice.

Detailed Description

The invention relates to an artificially cultivated phellinus igniarius sporocarp, wherein a phellinus igniarius strain, namely phellinus igniarius 1, is adopted as a mother strain of phellinus igniarius, the strain is preserved in China general microbiological culture collection center (CGCMCC) in 03 and 07 days 2014, the preservation number is CGCMCCNo.8854, and the strain is classified and named as phellinus igniarius Inonotus sanghuang and is disclosed in Chinese patent ZL 201410477690. X. Adopting a cultivation method of Phellinus linteus with imitating wild bagged material under natural forest (disclosed in Chinese patent 201711469522.6), selecting appropriate forest land and corresponding time for strain production, fungus stick production and forest land planting, and harvesting when Phellinus linteus fruiting body turns brown or dark brown from yellow.