CN107126450B - A Chinese medicinal preparation containing extract and effective substance, and its preparation method and application - Google Patents

Abstract

The invention discloses a koji grass extract, a koji grass effective substance enrichment, a preparation method and an application. The preparation method of the koji grass extract includes the following steps: mixing the koji grass with an ethanol aqueous solution and extracting to obtain an extract; wherein, the ethanol aqueous solution is 50% to 80% by volume. % ethanol in water. The sagebrush extract and sagebrush effective substance enrichment prepared by the invention can promote the excretion of uric acid and have stable drug effect, and further, the sagebrush effective substance enrichment can also reduce the level of urea nitrogen, and the effect on uric acid is reduced. Acute kidney injury has a certain protective effect; and Koji grass has abundant wild resources, low cost of medicinal materials, and great potential for development.

Description

Koji grass extract, Koji grass active substance enrichment, preparation method and application

technical field

The invention relates to a koji grass extract, a koji grass effective substance enrichment, a preparation method and an application.

Background technique

Uric acid is the final metabolite of human purine compounds, and disorders of purine metabolism can lead to hyperuricemia. Hyperuricemia (HUA) is defined as the fasting blood uric acid level higher than 420 μmol/L in men and higher than 360 μmol/L in women on two different days under a normal purine diet. Hyperuricemia is caused by increased production or decreased excretion of uric acid in the body. Hyperuricemia is the pathogenesis of gout. When the concentration of uric acid is supersaturated to form crystals and deposit in joint synovium, soft tissue, cartilage and kidney, it will cause "gout", thereby triggering the body's innate immune response, leading to joints and their surrounding areas. Tissue inflammation and renal impairment. In patients with hyperuricemia, the metabolic function of sugar and fat in the body will be significantly reduced, which is likely to cause various serious diseases, mainly diabetes, hypertension, hyperlipidemia, coronary heart disease, myocardial infarction, atherosclerosis, etc., which seriously threaten the human body. healthy.

With the improvement of people's living standards and the change of diet structure, the population of hyperuricemia in my country has shown an upward trend year by year. The demand for clinical uric acid-lowering drugs will increase year by year.

In Western medicine, uric acid-lowering therapy mainly controls uric acid levels by inhibiting uric acid production and increasing uric acid excretion or decomposition. At present, the chemical drugs or biological preparations used clinically for lowering uric acid mainly include the following categories:

(1) Inhibition of uric acid production-xanthine oxidase (XOD) inhibitors: allopurinol, febuxostat, etc.

(2) Promoting uric acid excretion-urate anion transporter (URAT1) inhibitors: benzbromarone, sulfinazolone, bebenecid, etc.

(3) Promote the decomposition of uric acid - uricases: Rasburicase, PEGylated recombinant uric acid oxidase (PEG-uricase).

There are clinical cases showing that long-term use of XOD inhibitor drugs, the recurrence rate of gout is still high. Long-term use of uricosuric drugs can cause severe hepatotoxicity and acute kidney injury, and professional physician guidance is especially required during drug use. At the end of 2014, CFDA has announced the adverse reaction notification of benzbromarone, reminding doctors and patients to be aware of the risks of drug use. Therefore, less toxic natural medicinal animal and plant resources have become an important source of raw materials for drug development.

Gnaphalium affine, (Gnaphalium affine, also known as Gnaphalium affine, Buddha's ear grass, white mugwort), is a plant of the Compositae family Gnaphalium affine. "Bielu of famous doctors" clearly records that "maintains cold, cold and heat"; "Pharmacopoeia" in 1977 records "dispels rheumatism and is used for rheumatic arthralgia". Before and after the Qingming Festival, people of all ethnic groups in many places in my country have the habit of picking their tender stems and leaves for consumption.

Literature Lin W, Xie J, Wu X, Yang L, Wang H.Inhibition of Xanthine OxidaseActivity by Gnaphalium Affine Extract. Chinese Medical Sciences Journal, 2014.29(4):225-230 discloses a kind of ethanolic extract of Koji grass (95 % ethanol extraction), and proved that it has the effect of inhibiting xanthine oxidase in vitro, that is, it has an inhibitory effect on the formation of uric acid.

SUMMARY OF THE INVENTION

The technical problem to be solved by the present invention is to overcome the defect that the existing uric acid excretion drugs have relatively large adverse reactions, and the existing uric acid production-inhibiting koji extract is not an ideal drug extract, and provides koji Grass extract, koji grass effective substance enrichment, preparation method and application. The sagebrush extract and sagebrush effective substance enrichment prepared by the invention can promote the excretion of uric acid and have stable drug effect, and further, the sagebrush effective substance enrichment can also reduce the level of urea nitrogen, and the effect on uric acid is reduced. Acute kidney injury has a certain protective effect; and Koji grass has abundant wild resources, low cost of medicinal materials, and great potential for development.

In the prior art, the 95% ethanol extract of Koji grass has a certain effect of inhibiting xanthine oxidase in vitro, that is, it has an inhibitory effect on the generation of uric acid. However, taking xanthine oxidase inhibitor drugs for a long time, the recurrence rate of gout is still relatively high. . From the perspective of disease relief mechanism, inhibiting the production of uric acid cannot effectively reduce the total amount of uric acid in the body from the root cause, especially when the total amount of uric acid is already at a high level, drugs that inhibit the production of uric acid are not an ideal solution. An ideal natural medicine extract for promoting uric acid excretion is an urgent problem to be solved, and finally, the present invention solves the above-mentioned technical problems through the following technical solutions.

The present invention provides a method for preparing a koji grass extract, which comprises the following steps: mixing koji grass with an ethanol aqueous solution, and extracting an extract; wherein, the ethanol aqueous solution is 50% by volume ~80% ethanol in water.

In the preparation method of the koji grass extract, the ethanol aqueous solution is preferably an ethanol aqueous solution with a volume percentage of 60%. The weight-to-volume ratio of the sage grass and the ethanol aqueous solution can be the ratio of the raw material to the solvent in the conventional phytochemical extraction, preferably 1:10～1:20kg/L, more preferably 1:10～1: 15kg/L. The sage grass is preferably dried in the shade. In this field, the shade-drying generally refers to placing the medicinal materials in a cool place indoors, outdoors or in a greenhouse, and using flowing air to blow off moisture to achieve the purpose of drying. "Generally refers to the sage grass with a mass content of about 4% moisture.

The extraction temperature can be 50-100°C, more preferably 70-80°C. The number of times of the extraction can be selected with reference to the routine in the art, and in the present invention, preferably 2 to 4 times, more preferably 3 times. The time for a single extraction in the extraction can be selected according to the routine in the field, and is preferably 1-2 hours in the present invention.

The preparation method of the Koji grass extract of the present invention may further include the step of concentrating the extract to obtain a concentrate or a concentrate.

Wherein, the conditions of the concentration can be selected with reference to the routine in the field, and this method is preferably concentrated under reduced pressure; the temperature of the concentration under reduced pressure is preferably 50-70°C. In the described "concentrating the extract to obtain a concentrate", the mass-volume ratio of the sage grass and the concentrate is preferably 1:0.5～1:2kg/L, more preferably 1:0.5～1 : 1kg/L. In the "concentrating the extract to obtain a concentrate", preferably the extract is concentrated to dryness to obtain the concentrate.

The present invention also provides a method for preparing the active substance enriched product of koji grass, which comprises the following steps: extracting the above-mentioned concentrated solution with petroleum ether to obtain water phase 1; extracting the water phase 1 with ethyl acetate to obtain water phase 2; The water-saturated n-butanol is extracted with the water phase II, and the water-saturated n-butanol phase is collected.

In the preparation method of the active substance enrichment of koji grass, the concentrated solution is generally the concentrated solution prepared by the preparation method of the koji grass extract, or can also be obtained by the The preparation method of Koji grass extract is obtained by mixing the prepared concentrate with water.

In the preparation method of described sage grass effective material enrichment, the consumption of described petroleum ether can be selected with reference to the routine in this area, in the present invention, the volume ratio of described concentrated solution and described petroleum ether It is preferably 1:1 to 1:2; the number of times of the petroleum ether extraction can be selected with reference to the routine in the art, and in the present invention, it is preferably 2 to 4 times, and more preferably 3 times.

The dosage of the ethyl acetate can be selected with reference to the routine in the field. In the present invention, the volume ratio of the water phase 1 to the ethyl acetate is preferably 1:1 to 1:2; the acetic acid The number of times of ethyl ester extraction can be selected with reference to the routine in the art, in the present invention, preferably 2 to 4 times, more preferably 3 times.

The water-saturated n-butanol can be the water-saturated n-butanol conventionally used in the field, and the configuration method is generally to mix n-butanol and distilled water at room temperature (0-40 degrees) (mixing volume ratio is 1:1), and vibrate. Shake, mix well, and stand overnight. The resulting mixture is layered, and the upper layer is a water-saturated n-butanol solution. The amount of the water-saturated n-butanol can be selected with reference to the routine in the field. In the present invention, the volume ratio of the water-saturated n-butanol to the water-saturated n-butanol is preferably 1:1 to 1:2; The number of times of the water-saturated n-butanol extraction can be selected with reference to the routine in the field, and in the present invention, preferably 2 to 4 times, more preferably 3 times.

In the preparation method of the active substance enrichment of koji grass, the steps of concentrating and drying the water-saturated n-butanol phase may be further included; wherein, the concentration is preferably reduced pressure concentration. The concentration is preferably by concentrating the water-saturated n-butanol phase to dryness.

The present invention also provides a koji grass extract prepared by the above-mentioned preparation method of the koji grass extract.

The present invention also provides a sagebrush effective substance enrichment prepared by the above-mentioned preparation method of the sagebrush effective substance enrichment.

The present invention also provides an application of the above-mentioned koji grass extract and/or koji grass effective substance enrichment in the preparation of a drug for promoting uric acid excretion.

The present invention also provides an application of the above-mentioned koji grass extract and/or koji grass active substance enrichment in the preparation of a medicine for treating and/or preventing related diseases caused by high uric acid level, or in the preparation of Application in functional food for regulating uric acid level.

The "related disorder caused by high uric acid level" in the present invention may be hyperuricemia and/or gout. The uric acid level-adjusting functional food generally refers to a functional food that reduces the uric acid level.

The present invention also provides a pharmaceutical composition, which comprises the above-mentioned koji grass extract and/or the above-mentioned koji grass effective substance enrichment, and a pharmaceutically acceptable excipient.

On the basis of not violating common knowledge in the art, the above preferred conditions can be combined arbitrarily to obtain preferred examples of the present invention.

The reagents and raw materials used in the present invention are all commercially available.

The positive and progressive effects of the present invention are: 1. Compared with similar chemical drugs, the product of the present invention is derived from Koji grass, with less toxic and side effects, and the wild resources of Koji grass are abundant, the cost of medicinal materials is low, and the development potential is huge; 2. This Compared with the 95% ethanol extract of Koji grass, the 50-80% ethanol extract of Koji grass in the invention has stronger uric acid excretion effect; and it is within a larger dose range (250-1000 mg/kg mice) It can basically maintain a good efficacy, so the stability of the efficacy is good; 3. The effective substance enrichment of koji grass can still maintain a significant efficacy when the dose is reduced by 4-8 times; it can be further Reducing blood urea nitrogen level has a certain protective effect on acute kidney injury caused by uric acid.

Detailed ways

The present invention is further described below by way of examples, but the present invention is not limited to the scope of the described examples. The experimental methods that do not specify specific conditions in the following examples are selected according to conventional methods and conditions, or according to the product description.

Example 1

After drying in the shade, the koji grass was cut into sections, weighed 1kg, mixed with 15L (1:15, kg/L) of 60% (v/v) ethanol aqueous solution, heated and refluxed for 2 times, 1h each time, and the extracts were combined. The extract was concentrated under reduced pressure (60° C.) and dried to obtain 166.4 g of a koji grass extract.

Example 2

After drying in the shade, the koji grass was cut into sections, weighed 5kg, mixed with 50% (v/v) aqueous ethanol solution 50L (1:10, kg/L, in a ratio, heated and refluxed for extraction 3 times, 2h each time, and merged the extract. Extraction. The liquid was concentrated under reduced pressure (70° C.), and dried to obtain 982.2 g of koji grass extract.

Example 3

After drying in the shade, koji grass was cut into sections, weighed 2kg, mixed with 40L (1:20, kg/L) of 80% (v/v) ethanol aqueous solution, heated and refluxed for 4 times, 2h each time, and the extracts were combined. The extract was concentrated under reduced pressure (50° C.), and dried to obtain 311.6 g of koji grass extract.

Example 4

After drying in the shade, the koji grass was cut into sections, weighed 5kg, mixed with 75L (1:15, kg/L) of 60% (v/v) ethanol aqueous solution, heated and refluxed for 4 times, 2h each time, and the extracts were combined. The extract is concentrated under reduced pressure (60° C.) to 5L (medicine: the volume of the concentrate is 1:1 kg/L) to obtain an extract concentrate.

The extraction concentrate was added with petroleum ether at 60-90°C for 3 extractions, the volume ratio of each concentrate and petroleum ether was 1:1 (v/v), the petroleum ether phase was discarded, and an aqueous phase was obtained. Add ethyl acetate to the aqueous phase, and extract for 3 times. Each time the volume ratio of aqueous phase 1 and ethyl acetate is 1:1 (v/v), and after extraction, aqueous phase 2 is obtained. Water-saturated n-butanol was added to the water phase 2, and extracted 3 times. The volume ratio of water-saturated n-butanol to water-saturated n-butanol was 1:1 (v/v) each time. 129.6g of sage grass effective substance enrichment.

Example 5

After drying in the shade, the koji grass was cut into sections, 0.6 kg was weighed, mixed with 6L (1:10, kg/L) of 50% (v/v) ethanol aqueous solution, heated and refluxed for 3 times, 2h each time, and the extracts were combined. The extract is concentrated under reduced pressure (70° C.) to 1.2L (medicine: the volume of the concentrate is 1:2kg/L) to obtain an extract concentrate.

The extraction concentrate was added with petroleum ether at 60-90°C to extract twice, the volume ratio of each concentrate and petroleum ether was 1:2 (v/v), the petroleum ether phase was discarded, and an aqueous phase was obtained. Add ethyl acetate to the water phase one, and extract twice, each time the volume ratio of water phase one to ethyl acetate is 1:2 (v/v), and after extraction, water phase two is obtained. Water-saturated n-butanol was added to the water phase 2, extracted twice, each time the volume ratio of water-saturated n-butanol to water-saturated n-butanol was 1:2 (v/v), the water-saturated n-butanol phases were combined, and the pressure was reduced to dryness to obtain Sage grass active substance enrichment 16.8g.

Example 6

After drying in the shade, koji grass was cut into sections, weighed 2kg, mixed with 40L (1:20, kg/L) of 80% (v/v) ethanol aqueous solution, heated and refluxed for 2 times, 2h each time, and the extracts were combined. The extract is concentrated under reduced pressure (50° C.) to 1L (medicine: the volume of the concentrate is 1:0.5kg/L) to obtain an extract concentrate.

The extraction concentrate was added with petroleum ether at 60-90° C. to extract four times, and the volume ratio of the concentrate to petroleum ether was 1:1 (v/v) each time, and the petroleum ether phase was discarded to obtain an aqueous phase 1. Add ethyl acetate to the aqueous phase, and extract for 4 times, each time the volume ratio of aqueous phase 1 and ethyl acetate is 1:1 (v/v), and after extraction, aqueous phase 2 is obtained. Water-saturated n-butanol was added to the water phase 2, and extracted 4 times. The volume ratio of water-saturated n-butanol to water-saturated n-butanol was 1:1 (v/v) each time. Sage grass active substance enrichment 49.6g.

In Comparative Example 1, the concentration of the aqueous ethanol solution was changed from 60% (v/v) to 95% (v/v), and the remaining conditions and steps were the same as those in Example 1.

Comparative Example 2, according to the existing literature "experimental study of Bai Ai extract on rats with hyperuricemia", Lin Weiqing, Xie Jianxiang, Wang Haidong, Chinese Journal of Rheumatology, 2005, 9(8), 509-510, No. Section 1.2.2, the prepared water extract of Koji grass.

Effect Example 1

Effect of the present invention Example 1 adopts the mouse hyperuricemia model caused by direct supplementation of uric acid, and the methods for preparation of the test substance, mode of administration, modeling, blood sample collection and the like are as follows:

Single-dose mode:

Rat species: ICR mice; Methods: Mice in the normal group and model group were given 0.5% sodium carboxymethyl cellulose aqueous solution (CMC-Na, g/mL) by gavage, 0.2 mL/10g body weight; mice in the administration group were given The CMC-Na aqueous suspensions of different concentrations of positive drugs or test substances (the extract of koji grass or the active substance enrichment of koji grass) were administered to the stomach, 0.2 mL/10g body weight. One hour later, the mice in the normal group were intraperitoneally injected with 1% CMC-Na aqueous solution (g/mL), and the model group and the administration group were intraperitoneally injected with uric acid suspension (1% CMC-Na aqueous solution, g/mL) , 0.2mL / mouse, the injection dose of 250mg uric acid / kg mouse body weight.

One hour after the injection of uric acid, 0.5 mL of blood was collected from the fundus venous plexus, stored in an ice bath, and centrifuged. The serum was passed through an automatic biochemical analyzer (Hitachi, Japan, model: 7080), uric acid (uricase method), urea nitrogen ( Urease-glutamate dehydrogenase method) detection kit to detect uric acid and urea nitrogen content.

The dosage specifications are as follows:

In the allopurinol group, allopurinol tablets (purchased from Shanghai Xinyi) were suspended in a 0.5% sodium carboxymethylcellulose aqueous solution (CMC-Na, g/mL) at a dose of 100 mg allopurinol/kg mouse body weight. Administration, administration volume 0.2mL/10g mouse body weight;

In the benzbromarone group, benzbromarone tablets (purchased from Hermann Pharmaceuticals, Germany) were suspended in a 0.5% sodium carboxymethylcellulose aqueous solution (CMC-Na, g/mL), and 50 mg of benzbromarone was added to the solution. Long/kg mouse body weight administration, the administration volume is 0.2mL/10g mouse body weight;

Uric acid suspension is uric acid (purchased from Sigma) suspended in 1% sodium carboxymethyl cellulose aqueous solution (CMC-Na, g/mL), the injection volume is 0.2mL / only, the injection dose is 250mg uric acid /kg mouse body weight.

Preparation of test substances (Examples 1-6 and Comparative Examples 1-3): Suspend the test substances (products prepared in Examples 1-6 and Comparative Examples 1-3) in 0.5% carboxymethyl In the sodium cellulose aqueous solution (CMC-Na, g/mL), the administration volume was 0.2 mL/10 g mouse body weight, and the specific administration dosages were shown in Tables 1-8.

Multiple dosing:

Rat species: ICR mice; Methods: Mice in the normal group and model group were given 0.5% sodium carboxymethyl cellulose aqueous solution (CMC-Na, g/mL) by gavage, each 0.2 mL/10 g body weight, for 5 consecutive times. day; the mice in the administration group were given the positive drug or the CMC-Na water suspension of different concentrations of the test substance (the extract of koji grass or the enriched substance of the effective substance) by gavage for 5 consecutive days. One hour after gavage on the last day, the mice in the normal group were intraperitoneally injected with 1% CMC-Na aqueous solution (g/mL), and the model group and the administration group were intraperitoneally injected with uric acid suspension (solvent is 1% CMC-Na aqueous solution, g/mL) 0.2mL/mice, the injection dose was 250mg uric acid/kg mouse body weight.

One hour after the injection of uric acid, 0.5 mL of blood was collected from the fundus venous plexus, stored in an ice bath, and centrifuged. The serum was passed through an automatic biochemical analyzer (Hitachi, Japan, model: 7080), uric acid (uricase method), urea nitrogen ( Urease-glutamate dehydrogenase method) detection kit to detect uric acid and urea nitrogen content.

The dosage specifications are as follows:

In the allopurinol group, allopurinol tablets (purchased from Shanghai Xinyi) were suspended in a 0.5% sodium carboxymethylcellulose aqueous solution (CMC-Na, g/mL) at a dose of 100 mg allopurinol/kg mouse body weight. Administration, administration volume 0.2mL/10g mouse body weight;

In the benzbromarone group, benzbromarone tablets (purchased from Hermann Pharmaceuticals, Germany) were suspended in a 0.5% sodium carboxymethylcellulose aqueous solution (CMC-Na, g/mL), and 50 mg of benzbromarone was added to the solution. Long/kg mouse body weight administration, the administration volume is 0.2mL/10g mouse body weight;

Uric acid suspension is uric acid (purchased from Sigma) suspended in 1% sodium carboxymethyl cellulose aqueous solution (CMC-Na, g/mL), the injection volume is 0.2mL / only, the injection dose is 250mg uric acid /kg mouse body weight.

Preparation of test substances (Examples 1-6 and Comparative Examples 1-3): Suspend the test substances (products prepared in Examples 1-6 and Comparative Examples 1-3) in 0.5% carboxymethyl In the sodium cellulose aqueous solution (CMC-Na, g/mL), the administration volume was 0.2 mL/10 g mouse body weight, and the specific administration dosages were shown in Tables 1-8.

The specific experimental data are shown in Tables 1-8:

Table 1. Single-dose uricosuric activity of the koji grass extract prepared in Example 1

Compared with the normal group: ^### P<0.001; compared with the model group *P<0.05, **P<0.01

The experimental results in Table 1 show that the koji grass extract prepared in Example 1 has a significant uricosuric effect at a dose of 500 mg/kg, compared with the model group, after a single administration. Compared with the model group, there was no significant difference in blood uric acid level after single administration of the koji grass extract (500 mg/kg) prepared in Comparative Example 1 and Comparative Example 2, that is, there was no obvious uricosuric effect.

Table 2. Multi-administration uricosuric activity of the koji grass extract prepared in Example 1

Compared with normal group: ^### P<0.001; compared with model group *P<0.05, **P<0.01, ***P<0.001

The experimental results in Table 2 show that the koji grass extract of Example 1 at doses of 250, 500 and 1000 mg/kg, after multiple administrations, has a significant uricosuric effect compared with the model group. Compared with the model group, the water extracts (500mg/kg) prepared by Comparative Example 1 and Comparative Example 2 showed a certain effect of promoting uric acid excretion, but P<0.05. It is still weaker than the koji grass extract prepared in Example 1.

Table 3. Single-dose uricosuric activity of koji grass extract prepared in Example 2

Compared with the normal group: ^### P<0.001; compared with the model group **P<0.01

The experimental results in Table 3 show that the koji grass extract prepared in Example 2 has a significant uricosuric effect compared with the model group after a single dose of 500 and 1000 mg/kg. Compared with the model group, the water extract (500 mg/kg) prepared in Comparative Example 2 showed no significant difference, and the effect of promoting uric acid excretion was not obvious after a single administration.

Table 4. Multi-dose uricosuric activity of the koji grass extract prepared in Example 2

Compared with normal group: ^### P<0.001; compared with model group *P<0.05, **P<0.01, ***P<0.001

The experimental results in Table 4 show that the 250, 500, and 1000 mg/kg doses of the koji grass extract in Example 2 have a significant uricosuric effect compared with the model group after repeated administration. Compared with the model group, the water extract (500mg/kg) prepared in Comparative Example 2 showed a certain effect of promoting uric acid excretion after multiple administrations, P<0.05, but the activity was weaker than that of the mice in Example 2. Koji grass extract.

Table 5 Multi-administration uricosuric activity of the koji grass extract prepared in Example 3

Compared with the normal group: ^### P<0.001; compared with the model group *P<0.05, **P<0.01

The experimental results in Table 5 show that the koji grass extract prepared in Example 3 at doses of 250, 500, and 1000 mg/kg, after repeated administration, has a significant uricosuric effect compared with the model group. Compared with the model group, the water extract (500mg/kg) prepared in Comparative Example 2 showed a certain uric acid excretion effect, but the activity was weaker than that of the mice prepared in Example 3. Koji grass extract.

Table 6 Multi-administration uricosuric activity of the active substance enrichment of koji grass prepared in Example 4

Compared with normal group: ^## P<0.01, ^### P<0.001; compared with model group *P<0.05, **P<0.01, ***P<0.001

The experimental results in Table 6 show that the active substance enrichment of koji grass prepared in Example 4 at doses of 62.5, 125, and 250 mg/kg, after repeated administration, has a significant uricosuric excretion effect compared with the model group; and The 62.5 and 125 mg/kg dose levels can significantly reduce the urea nitrogen level, the activity is higher than that of the benzbromarone group, and it has a certain protective effect on acute kidney injury caused by uric acid.

Table 7 Multi-administration uricosuric activity of the active substance enrichment of koji grass prepared in Example 5

Compared with the normal group: ^### P<0.001; compared with the model group *P<0.05, **P<0.01

The experimental results in Table 7 show that the active substance enrichment of koji grass prepared in Example 5 at doses of 62.5, 125, and 250 mg/kg, after repeated administration, has a significant uricosuric excretion effect compared with the model group; and At the dose levels of 62.5 and 125 mg/kg, the level of urea nitrogen can be significantly reduced, and the activity is significantly higher than that of the benzbromarone group, which has a certain protective effect on acute kidney injury caused by uric acid.

Table 8 Multi-administration uricosuric activity of the active substance enrichment of koji grass prepared in Example 6

Compared with normal group: ^## P<0.01, ^### P<0.001; compared with model group *P<0.05, **P<0.01

The experimental results in Table 8 show that the active substance enrichment of koji grass prepared in Example 6 at doses of 62.5, 125, and 250 mg/kg, after multiple administrations, compared with the model group, all have significant uricosuric effect; And at the dose levels of 62.5, 125, and 250 mg/kg, the level of urea nitrogen was significantly different from that of the model group, which could significantly reduce the level of urea nitrogen, and its activity was significantly higher than that of the benzbromarone group, which had a certain protective effect on acute kidney injury caused by uric acid. .

Claims (15)

1. A preparation method of an aspergillus oryzae extract is characterized by comprising the following steps: mixing herba Gnaphalii affinis with ethanol water solution, and extracting to obtain extractive solution; wherein the ethanol water solution is 50-80% by volume; the weight volume ratio of the aspergillus oryzae to the ethanol aqueous solution is 1: 10-1: 20 kg/L; the said mouse-rice-koji grass is dried in the shade; the extraction temperature is 50-100 ℃; the extraction times are 2-4 times; the time of single extraction in the extraction is 1-2 h.

2. The method of preparing Aspergillus oryzae extract of claim 1, wherein the extract is obtained by subjecting Aspergillus oryzae to a reaction,

the ethanol water solution is 60 percent ethanol water solution by volume percentage;

and/or the weight volume ratio of the aspergillus oryzae to the ethanol aqueous solution is 1: 10-1: 15 kg/L;

and/or the extraction temperature is 70-80 ℃;

and/or, the extraction times are 3 times;

and/or, the preparation method of the aspergillus oryzae extractive also comprises the step of concentrating the extracting solution to obtain concentrated solution or concentrate; the concentration is reduced pressure concentration; the temperature of the reduced pressure concentration is 50-70 ℃.

3. The method of preparing Aspergillus oryzae extract as claimed in claim 2, wherein the extract is prepared by extracting Aspergillus oryzae,

in the step of concentrating the extracting solution to obtain a concentrated solution, the mass volume ratio of the aspergillus oryzae to the concentrated solution is 1: 0.5-1: 2 kg/L;

the step of concentrating the extracting solution to obtain the concentrate is to concentrate the extracting solution to be dry to obtain the concentrate.

4. The method of claim 3, wherein the mass/volume ratio of the mouse-koji grass to the concentrated solution is 1:0.5 to 1:1 kg/L.

5. A mouse-koji grass extract obtained by the method for preparing a mouse-koji grass extract according to any of claims 1 to 4.

6. A preparation method of an effective substance enrichment substance of a koji grass is characterized by comprising the following steps: extracting the concentrated solution according to any one of claims 2 to 4 with petroleum ether to obtain a first aqueous phase; extracting the water phase I with ethyl acetate to obtain a water phase II; extracting the water phase II with water saturated n-butanol, and collecting the water saturated n-butanol phase.

7. The process for producing an enriched product of an effective substance derived from mouse koji grass according to claim 6,

in the preparation method of the aspergillus oryzae effective substance enrichment, the volume ratio of the concentrated solution to the petroleum ether is 1: 1-1: 2; and/or the petroleum ether is extracted for 2-4 times;

and/or the volume ratio of the water phase I to the ethyl acetate is 1: 1-1: 2; and/or the extraction times of the ethyl acetate are 2-4 times;

and/or the volume ratio of the water phase II to the water-saturated n-butanol is 1: 1-1: 2; and/or the extraction times of the water saturated n-butanol are 2-4 times.

8. The method of claim 7, wherein the number of petroleum ether extractions is 3.

9. The method of claim 7, wherein the number of ethyl acetate extractions is 3.

10. The process of claim 7, wherein the extraction with water-saturated n-butanol is carried out 3 times.

11. The method of any of claims 6-10, further comprising concentrating and drying the water-saturated n-butanol phase; the concentration is reduced pressure concentration; the concentration is to concentrate the water saturated n-butanol phase to dryness.

12. An effective substance enrichment of mouse-koji grass prepared by the method of any of claims 6 to 11.

13. Use of the extract of mouse-koji grass according to claim 5 and/or the concentrate of the effective substances of mouse-koji grass according to claim 12 for the preparation of a medicament for promoting uric acid excretion.

14. Use of an Aspergillus oryzae extract as defined in claim 5 and/or an Aspergillus oryzae effective substance concentrate as defined in claim 12 for the preparation of a medicament for the treatment and/or prevention of disorders related to high uric acid levels.

15. A pharmaceutical composition for reducing uric acid, comprising the koji grass extract of claim 5 and/or the koji grass effect substance concentrate of claim 12, and a pharmaceutically acceptable excipient.