阅读说明：本技术 一种用于治疗高尿酸及其相关疾病的雀嘴茶提取物及其制备方法、制剂与应用 (Tea extract of sparrow mouth for treating hyperuricemia and related diseases, and preparation method, preparation and application thereof ) 是由 丛力 高志奎 任杨帆 沈报春 尹开云 马彦 于 2021-07-30 设计创作，主要内容包括：本发明涉及一种用于治疗高尿酸及其相关疾病的雀嘴茶提取物及其制备方法、制剂与应用,属于生物医药技术领域。本发明采用雀嘴茶顶尖上新生嫩叶作为原料,经过提取、结晶、干燥等工艺,所得到的雀嘴茶提取物中6’-O-咖啡酰熊果苷质量含量大于85%,对苯二酚质量含量小于0.08%。该雀嘴茶提取物可以在制备治疗、预防高尿酸及其相关疾病药物中应用。经科学研究和多年的临床研究证实,本发明以富含6’-O-咖啡酰熊果苷的雀嘴茶提取物对高尿酸及其并发症有较好的治疗作用,尤其对高尿酸结石引起的肾衰竭有很好的治愈功效,能够在30天左右治疗周期很好地排出肾积水,显著降低肌酐,并明显改善患者的生存质量。(The invention relates to a sparrow tea extract for treating hyperuricemia and related diseases, and a preparation method, a preparation and application thereof, and belongs to the technical field of biological medicines. The method adopts the newly-born tender leaves on the tip of the tea leaves of the sparrow mouth as raw materials, and the 6' -O-caffeoyl arbutin mass content in the obtained extract of the tea leaves of the sparrow mouth is more than 85 percent and the hydroquinone mass content is less than 0.08 percent through the processes of extraction, crystallization, drying and the like. The tea extract can be used for preparing medicines for treating and preventing hyperuricemia and related diseases. Scientific research and years of clinical research prove that the Quezui tea extract rich in 6' -O-caffeoyl arbutin has better treatment effect on hyperuricemia and complications thereof, particularly has good curative effect on renal failure caused by hyperuricemia, can well discharge hydronephrosis in a treatment period of about 30 days, remarkably reduces creatinine, and obviously improves the life quality of patients.)

1. The sparrow tea extract for treating hyperuricemia and related diseases is characterized in that the mass content of 6' -O-caffeoyl arbutin in the sparrow tea extract is more than 85%, and the mass content of hydroquinone is less than 0.08%.

2. The tea finch extract for treating hyperuricemia and related diseases according to claim 1, further comprising 0.5% -15% by mass of a phenolic compound, wherein the phenolic compound comprises at least one of ouabain A, caffeic acid and arbutin.

3. The method for preparing the extract of Quezetima for treating hyperuricemia and related diseases according to claim 1 or 2, comprising the steps of:

step (1), selecting raw materials: selecting new young leaves of the maidenhair tea in the months of 4-8, and cleaning the fresh leaves with tap water after picking;

step (2), extraction: extracting the raw materials treated in the step (1) in an iron container; the time from picking to extraction is not more than 12 h;

the specific extraction method comprises the following steps: adding drinking water to cover the raw materials, boiling with strong fire for 30-40 min, cooling to 50 + -10 deg.C, further grating the leaves, cooling to 40 + -10 deg.C, filtering, cooling the filtrate to 30 + -10 deg.C, stirring for 30-40 min, stopping stirring when white substance appears, and standing in dark for crystallization;

and (3) post-processing: and removing the supernatant of the crystallization solution under the conditions of keeping out of light and heat, filtering or centrifuging, taking the crystal, washing the crystal with water at 4-10 ℃, and drying the crystal in vacuum to obtain the griffonia simplicifolia extract.

4. The preparation method of the tea leaf extract of sparrow beak for treating hyperuricemia and related diseases according to claim 3, wherein if the tea leaf cannot be extracted in time within 12 hours, the picked tea leaf is first subjected to de-enzyming treatment, then dried at 55-65 ℃ to obtain a dry product, and then pulverized or cut into 5-10 mesh coarse powder to be directly used as an extraction medicinal material.

5. The method for preparing the extract of Quezui tea for treating hyperuricemia and related diseases according to claim 3, further comprising a refining step; the refining step is 1 or more times;

adding water with the mass 2-3 times of that of the crystal into the washed crystal under a dark inert gas, heating and refluxing the crystal on a steam bath until the crystal is completely dissolved, then cooling the crystal to 75-85 ℃, then adding activated carbon for refluxing and decoloring, filtering, naturally cooling the filtrate for crystallization, filtering, washing the crystal with water with the temperature of 5-15 ℃, and then drying in vacuum to obtain the refined product of the maidenhair tea extract.

6. The method for preparing the extract of hippophae rhynchophylla for treating hyperuricemia and related diseases according to claim 5, wherein the mass ratio of the activated carbon to the washed crystal is 10-50: 1000, parts by weight; the specific method for vacuum drying comprises the following steps: vacuum drying is carried out for 10-14 h at the temperature of 60 +/-5 ℃ and under the mmHg of 80-100 mmHg.

7. A pharmaceutical preparation containing the tea leaves extract of the sparrow beak for treating hyperuricemia and related diseases according to claims 1-2, which is characterized by comprising an active ingredient and auxiliary materials; the active component comprises a tea extract of the bird's beak; the dosage forms of the pharmaceutical preparation comprise tablets, granules, capsules, paste, oral liquid, intramuscular injection and intravenous injection.

8. The pharmaceutical preparation containing the extract of Quezetima for treating hyperuricemia and related diseases according to claim 7, wherein:

when the pharmaceutical preparation is granules, capsules or tablets, the pharmaceutical preparation is composed of the following raw materials by weight ratio, and comprises, by 1000 granules or tablets, 100-120 g of the sparrow tea extract, 22-38 g of dextrin, 9-11 g of microcrystalline cellulose, 4.7-5.3 g of plantain seed gum, 9-11 g of konjac polysaccharide, 9-11 g of sodium carboxymethyl starch and 1.9-1.1 g of magnesium stearate;

when the medicinal preparation is a beverage, the medicinal preparation consists of the following raw materials in percentage by weight, and the raw materials are counted by 1000 bottles and 10-200 ml/bottle: 50-170 g of a tea extract of the bird's beak, 39.7-40.3 g of plantain seed gum, 59-61 g of konjac polysaccharide, 3-7 g of stevioside and the balance of water;

when the medicinal preparation is a freeze-dried powder injection, the medicinal preparation consists of the following raw materials in percentage by weight, and the raw materials are counted in 1000 bottles and 5-10 ml/bottle: 50-150 g of the sparrow tea extract, 40-60 g of mannitol and water for injection to the total amount.

9. The use of the tea extract of Quezui for treating hyperuricemia and related diseases according to claims 1-2 for preparing a medicament for treating and preventing hyperuricemia and related diseases.

10. The use of the extract of Kuckunzu tea for treating hyperuricemia and related diseases according to claim 9, wherein the hyperuricemia and related diseases include hyperuricemia, gout, hyperuricemia, nephropathy with hyperuricemia, renal calculus with hyperuricemia, and cardiovascular and cerebrovascular diseases with hyperuricemia.

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to a sparrow tea extract for treating hyperuricemia and related diseases, a preparation method, a preparation and an application thereof, in particular to an application of the sparrow tea extract rich in 6' -O-caffeoyl arbutin in preparation of medicines for treating hyperuricemia and related diseases.

Background

Human beings have had plenty of lives because of rapid development, but have come with various chronic diseases such as modern diseases and diseases with great value. In addition to hypertension, hyperglycemia, and hyperlipidemia, hyperuricemia has been receiving attention in recent years. Gout is only known to be a mysterious disease and is a affluent disease, the main cause of gout is hyperuricemia, and 10% of patients with hyperuricemia can further develop into gout. And gout has become a common disease with a morbidity rate of up to 1% from mysterious diseases of a few people. There is a growing body of evidence that hyperuricemia is not only a major cause of gout, but also a major cause of cardiovascular and cerebrovascular diseases, and also a major risk factor of arthritis and nephropathy. Such diseases are also known as hyperuricemia-related diseases (or complications).

Although the harm of high uric acid has been increasingly paid attention to the medical field, uric acid is not only an inevitable product of human metabolism, but also is an important antioxidant in vivo, and has important physiological effects. In view of the importance of uric acid and the health problems caused by excessive uric acid, the medicine world is actively developing, and the clinically considered better medicines comprise allopurinol, benzbromarone and the like, the new medicine febuxostat and the like, and colchicine which is a famous-brand medicine for treating acute gout. These chemicals have more adverse reactions and are more serious, and some of them, such as benzbromarone, have been cancelled (forbidden) in many countries. Even the new drug febuxostat has been black boxed by the FDA in the united states to warn of cardiovascular and cerebrovascular risk.

While the safety of the chemical drugs is concerned, the traditional drugs are also concerned, and many flavonoids, polyphenols, cellulose and the like in the plants are concerned and proved to be effective. For example, mangiferin aglycon is a strong XOD inhibitor, and mangiferin, apigenin and the like have published papers and applied for patents on research of high uric acid and gout. Although many researches and reports are carried out so far, products with a known curative effect on reducing uric acid are still few, and the chemical febuxostat tablet is a good product with a good known effect and acceptable safety. The society needs safer and more effective uric acid reducing drugs!

As a natural plant Quezui tea, the folk says that the Quezui tea is used as a tribute and has various health-care effects of clearing heat and removing toxicity and the like. However, researchers also indicate that the medicine has the defects of potential safety hazard, unstable quality and effect, unobvious uric acid reducing effect and the like. The active ingredients of the sparrow tea are also researched and reported more, and the data of the prior patent applications and technical publications show that: zhang Yinjun, et al, a Kunming plant institute of Kunming, academy of sciences in CN103120624A and CN101085792A, disclose a maidenhair tea extract, which has been studied for whitening efficacy of components contained therein. The Yunnan Baiyao group Zhao Sanren and the university of Huaqiao also have the research on the chemical components in the broom-mouth tea. The safety problem of the plum chrysanthemum and the like is shown by the preliminary study on the median lethal dose and pathology of the bird's beak tea. Related published patent applications are: CN111939189A, CN111939190A, CN111533770A and the like.

The extraction methods disclosed in patents CN111471078A and CN1153370A use ethanol for extraction, ethyl acetate for extraction, and column chromatography, which is complicated in process and difficult in mass industrial production; and the organic solvent is adopted, the content of the main components and the content of toxic components in the extract are not determined,

in studies on chemical components of plants in the genus Vaccinium, Zao et al isolated the following compounds 6-O-Acetylarbutin (6-O-Acetylarbutin), Arbutin (Arbutin), Avicularin (Avicillarin), 2-O-caffeoyl Arbutin (2-O-caffeoyl Arbutin), (-) -epicatechin (epicatechin), Hydroquinone (Hydroquinone), 4-Hydroxyphenyl-beta-gentiobioside (4-Hydroxyphenyl-beta-gentiobioside), indigo honeysuckle A2(proanthocyanidin A2), Salidroside (Salidroside), echinomycin-3-O-beta-D-galactoside (cyanodin-3-O-beta-D-galactoside).

As can be known from chemical safety handbooks and literature reports, the relative toxicity of the main active phenolic components in the tea leaves of the sparrow mouths is as follows: hydroquinone (hydroquinone) > arbutin > 6' -O-caffeoyl arbutin (only 1/2 for arbutin toxicity). Hydroquinone is a white crystal, also called hydroquinone. It is toxic, and 1g of the medicine is taken by adults by mistake, and symptoms such as headache, dizziness, tinnitus, pale complexion and the like can appear. Therefore, the toxic existence of the active ingredients of the tea leaves is beneficial to the use desire and research and development enthusiasm of people, and the tea leaves are one of the important reasons for the better development and utilization of the tea leaves. How to control the toxicity of the active ingredients in the tea leaves and exert the efficacy of the active ingredients in the tea leaves of sparrow mouths needs to be solved from the source and the technical aspect.

Disclosure of Invention

In order to solve the defects of the prior art, the first purpose of the invention is to provide a low-toxicity and safe Quezui tea extract which is rich in 6' -O-caffeoyl arbutin and is used for treating hyperuricemia and related diseases; the second purpose is to provide a preparation method for reducing the toxicity of the tea extract; the third object is to provide a pharmaceutical preparation containing the extract of Quezetima latifolia rich in 6' -O-caffeoyl arbutin; the fourth purpose is to provide the application of the sparrow tea extract in preparing the medicines for treating hyperuricemia and related diseases, in particular to the application in preparing the medicines for improving renal function.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

a tea extract of fructus Canarii albi for treating hyperuricemia and related diseases contains 6' -O-caffeoyl arbutin more than 85 wt% and hydroquinone less than 0.08 wt%.

Further, the tea drink also preferably contains 0.5-15% of phenolic compounds by mass, wherein the phenolic compounds comprise at least one of oujin glycoside A, caffeic acid and arbutin.

The structural formula of the ouabain A is as follows:

Robustaside A (Wujingan A)

The structural formula of caffeic acid:

the structural formula of arbutin is:

the invention also provides a preparation method of the tea extract of the sparrow tea for treating hyperuricemia and related diseases, which comprises the following steps:

step (1), selecting raw materials: selecting new young leaves of the maidenhair tea in the months of 4-8, and cleaning the fresh leaves with tap water after picking;

step (2), extraction: extracting the raw materials treated in the step (1) in an iron container; the time from picking to extraction is not more than 12 h;

the specific extraction method comprises the following steps: adding drinking water to cover the raw materials, boiling with strong fire for 30-40 min (based on the boiling of the leaves), cooling to 50 + -10 deg.C, further rubbing the leaves, cooling to 40 + -10 deg.C, filtering, cooling the filtrate to 30 + -10 deg.C, stirring for 30-40 min, stopping stirring when white substance appears, and standing in dark for crystallization;

and (3) post-processing: and removing the supernatant of the crystallization solution under the conditions of keeping out of light and heat, filtering or centrifuging, taking the crystal, washing the crystal with water at 4-10 ℃, and drying the crystal in vacuum to obtain the griffonia simplicifolia extract.

HPLC chromatograms of Quezui tea and hydroquinone control are shown in FIG. 1. HPLC chromatogram of 6' -O-caffeoyl arbutin, caffeic acid and oujin glycoside A is shown in FIG. 2; precisely weighing 0.5g of the sparrow beak tea extract, diluting the volume of methanol to 25.00mL, filtering with a 0.45 mu m microporous filter membrane, carrying out sample injection analysis, and obtaining a chromatogram shown in figure 3.

The chromatographic conditions of FIG. 2 were: a chromatographic column: 20RB-AX SB-C18(9.4 x 250mm, 5 μm); shimadzu LC-2010AHT high performance liquid chromatograph, mobile phase: methanol-water gradient elution, conditions are shown in table 1, flow rate: 3.0ml/min, column temperature: 30 ℃, detection wavelength: 254nm, sample size: 10 μ l.

TABLE 1 gradient elution schedule

Time	Methanol
		0-40min	30-70％
40-50min	65-35％

The chromatographic conditions of FIG. 3 were: a chromatographic column: agilent ZOBXa-C18 column (4.6X 250mm, 5 μm); shimadzu LC-2010A high performance liquid chromatograph, mobile phase: methanol-1% acetic acid water, gradient elution procedure as shown in table 2, flow rate: 1.0mL/min, column temperature: 25 ℃, detection wavelength: 280nm, sample injection amount: 2 μ L.

TABLE 2 HPLC gradient elution procedure

Time	Methanol
		0-10min	25-30％
10-15min	30-34％
		15-20min	34％
20-27min	34-40％
		27-32min	40-100％

Two pure products with main components are obtained by preparative liquid chromatography, and are determined to be 6' -O-caffeoyl arbutin (t) by NMR analysis and comparison with literature_R19.317) and ouabain A (t)_R27.562). Both accounted for 97.3% of the samples.

Note: when the gradient elution is performed, the elution ratio is linearly changed.

Further, preferably, if the tea cannot be extracted within 12 hours in time, the picked tea is subjected to enzyme deactivation treatment, then dried at the temperature of 55-65 ℃ to form a dry product, and then crushed or cut into coarse powder of 5-10 meshes to be directly used as an extracted medicinal material.

Further, it is preferable to further include a refining step; the refining step is 1 or more times;

and adding water with the mass 2-3 times that of the crystal into the washed crystal in a dark inert atmosphere, heating and refluxing the crystal on a steam bath until the crystal is completely dissolved, cooling the crystal to 75-85 ℃, adding activated carbon, refluxing and decoloring, filtering, naturally cooling the filtrate for crystallization, filtering, washing the crystal with water at the temperature of 5-15 ℃, and then drying in vacuum to obtain the refined product of the maidenhair tea extract.

Further, it is preferable that the mass ratio of the activated carbon to the washed crystal is 10 to 50: 1000, parts by weight; the specific method for vacuum drying comprises the following steps: vacuum drying is carried out for 10-14 h at the temperature of 60 +/-5 ℃ and under the mmHg of 80-100 mmHg.

The invention also provides a pharmaceutical preparation containing the tea extract for treating the diseases related to the hyperuricemia, which comprises active ingredients and auxiliary materials; the active component comprises a tea extract of the bird's beak; the dosage forms of the pharmaceutical preparation comprise tablets, granules, capsules, paste, oral liquid, intramuscular injection and intravenous injection.

Further, preferably, when the pharmaceutical preparation is granules, capsules or tablets, the pharmaceutical preparation is composed of the following raw materials by weight ratio, based on 1000 granules or tablets, 100-120 g of the broadleaf holly leaf extract, 22-38 g of dextrin, 9-11 g of microcrystalline cellulose, 4.7-5.3 g of plantain seed gum, 9-11 g of konjac polysaccharides, 9-11 g of sodium carboxymethyl starch and 1.9-1.1 g of magnesium stearate;

when the medicinal preparation is a beverage, the medicinal preparation consists of the following raw materials in percentage by weight, and the raw materials are counted by 1000 bottles and 10-200 ml/bottle: 50-170 g of a tea extract of the bird's beak, 39.7-40.3 g of plantain seed gum, 59-61 g of konjac polysaccharide, 3-7 g of stevioside and the balance of water;

when the medicinal preparation is a freeze-dried powder injection, the medicinal preparation consists of the following raw materials in percentage by weight, and the raw materials are counted in 1000 bottles and 5-10 ml/bottle: 50-150 g of the sparrow tea extract, 40-60 g of mannitol and water for injection to the total amount.

The invention also provides application of the tea extract for treating hyperuricemia and related diseases in preparation of medicines for treating and preventing hyperuricemia related diseases.

Further, it is preferable that the hyperuricemia and related diseases include hyperuricemia, gout, hyperuricemia arthritis, hyperuricemia nephropathy, hyperuricemia renal calculus, and hyperuricemia cardiovascular and cerebrovascular diseases.

The extraction equipment only needs iron equipment, and the extraction method is a whole-process water extraction method (green and environment-friendly).

According to the invention, the sparrow tea extract can be obtained by storing the dried product for later use as a raw material and then processing the raw material according to the methods from the step (2) to the step (5).

In step (2) of the present invention, the filtration means of the present invention is not particularly limited, and for example, filtration using a coarse filter cloth, the resulting filtrate is usually red. The time for crystallization is not particularly limited, as the case may be, for example, 24 hours. The stirring speed is not particularly limited, and the stirring is slow, for example, 10 to 20 r/min.

The crystal obtained after filtration is usually washed after liquid is drained; the water used for washing is preferably pure water, and when washing the crystal, a filtration method is preferably used, and filtration is performed while adding pure water, but the present invention is not limited thereto.

In the refining step, the adopted water is preferably distilled water, and the reflux decoloring time is preferably 10-30 min, but the method is not limited to this. In the refined product of the Quezui tea extract, the content of 6' -O-caffeoyl arbutin is more than 99.0 percent, and the content of hydroquinone is not more than 0.02 percent.

The inert gas atmosphere in the present invention is preferably a nitrogen atmosphere.

The tea extract of the invention can be used as an effective component in the preparation of drugs for treating diseases related to uric acid. The tea extract of the sparrow beak is applied to the preparation of anti-inflammatory, bacteriostatic, antioxidant and uric acid-reducing medicines for treating hyperuricemia and complications.

The auxiliary materials in the invention are medically acceptable auxiliary materials.

When the pharmaceutical preparation is granules, capsules or tablets, the sparrow tea extract, dextrin, microcrystalline cellulose, plantain seed gum, konjac polysaccharide and sodium carboxymethyl starch are weighed according to the prescription amount, the mixture is placed in a wet granulator and mixed for 10 +/-1 min, ethanol with the volume concentration of 50 percent is added, and granulation process parameters are set (the stirring speed is 700 r.min)^-1The cutting speed is 1000 r.min^-1Granulating for 5min), drying, controlling the water content of the granules to be 4-6%, sieving with a 20-mesh sieve, grading, adding magnesium stearate into the granules, mixing uniformly, and then filling into No. 1 capsules to obtain capsules; the granules were compressed to give tablets. The daily dose (based on the content of the extract) of the adult is not more than 5-8g, and the adult takes the medicine for three times. Is suggested as a medicine for treating hyperuricemia and related diseases.

When the medicinal preparation is a beverage, weighing the tea extract, the plantain seed gum, the konjac polysaccharide and the stevioside according to the prescription amount, mixing, adding water to the prescription amount, dissolving, subpackaging according to 1000 bottles, and sterilizing to obtain the standard extract beverage of the tea extract; the daily dose (based on the content of the extract) of the adult is not more than 3-6 g, and the adult takes the medicine for three times. Is suggested as a medicine for treating hyperuricemia and related diseases.

When the medicinal preparation is a freeze-dried powder injection, weighing the sparrow tea extract and mannitol according to the prescription amount under the condition of cGMP, adding injection water to 80% of the total amount of the injection water, heating to 45-50 ℃, adding the rest injection water after dissolving, stirring uniformly, performing rough filtration of 0.45 mu m, performing fine filtration of 0.2 mu m, filling 2ml of liquid medicine in a 5ml penicillin bottle, and performing freeze vacuum drying to obtain the freeze-dried powder injection for injection; the daily dose (calculated according to 6' -O-caffeoyl arbutin) for adults is not more than 1-2 g, and the daily dose is used by three injections. Is suggested as a medicine for treating gout or acute hyperuricemia and renal failure.

The invention relates to an application of a sparrow mouth tea extract in preparing a medicine for treating and preventing gout or hyperuricemia and related complications. Specifically, the tea extract of the bird's beak is used as follows:

(1) the application in preparing the medicinal preparation for treating gout or hyperuricemia;

(2) the application in preparing the medicine for preventing hyperuricemia or inhibiting the further generation of hyperuricemia;

(3) the application in preparing the medicine for treating gout or uric acid calculi;

(4) the application in preparing the medicine for treating renal failure, namely the application in preparing the medicine for treating hyperuricemia, gout, hyperuricemia (gouty) arthritis, hyperuricemia nephropathy, hyperuricemia renal calculus and hyperuricemia cardiovascular and cerebrovascular diseases;

(5) the application in preparing the medicine for treating hyperuricemia-induced hypercreatinine nephropathy in uric acid related diseases;

the application of the tea extract of the sparrow beak in preparing the medicine for treating gout or hyperuricemia can prepare products which are beneficial to improving the symptoms of uric acid patients, and the products comprise food, tea, beverage, health-care food, hospital preparation, medical nutriment and medicine.

The specific dosage of the tea extract for treating diseases related to hyperuricemia is 0.5g-8 g/day.

The invention provides an integrated and innovative therapeutic product for hyperuricemia and related diseases and curative effect characterization, and provides a promising product for the medical problem, namely renal failure.

The applicant researches and discovers that in the processing and extraction processes of the sparrow tea after picking, if the processing is improper, the main effective component (6' -O-caffeoyl arbutin or called oujin glycoside A) is easy to metabolize or hydrolyze into arbutin with higher toxicity, and then further metabolize or hydrolyze into hydroquinone with higher toxicity. The key reason for the toxicity of the sparrow tea extract in the research of the prior art is that the sparrow tea extract is considered to be the key problem to be solved urgently in the prior art. The invention fundamentally solves the problem, and experimental research proves that the detoxified sparrow tea extract has very good clinical effect on preparing the medicament for treating gout and improving kidney function, and toxicity tests prove that the detoxified sparrow tea extract is safe, effective and controllable.

The invention is directly extracted by water, and the mass content of 6' -O-caffeoyl arbutin in the extract is determined to be not less than 85 percent, and the mass content of the toxic component hydroquinone (hydroquinone) is determined to be less than 0.08 percent; animal experiments show that the medicine has the effect of reducing uric acid and urine protein, and has positive significance for treating kidney disease and kidney failure; the invention evaluates the curative effect of a plurality of gout and hyperuricemia patients, and finds that the invention has better curative effect.

In the invention, 6' -O-caffeoyl arbutin, also called wujin glucoside, is a main active ingredient; hydroquinone, also known as hydroquinone, is the major toxic component.

The research finds that the reason for the toxicity generation (increase) of the sparrow tea extract is as follows: arbutin and its derivatives can be continuously hydrolyzed (degraded) to produce hydroquinone which is the final degradation product, and hydroquinone can be continuously oxidized into quinone and other polymers.

The metabolic pathway of the ingredients in plants or the hydrolysis pathway in the extraction process is wujin glycoside → arbutin → hydroquinone.

In the process of post-picking treatment and extraction, if the treatment is improper, the main component (6' -O-caffeoyl arbutin or oujin glycoside A) is easy to metabolize or hydrolyze into arbutin with higher toxicity, and then further metabolize or hydrolyze into hydroquinone with higher toxicity. This is a very critical issue. The technical scheme solves the problem effectively through two innovative and effective schemes, so that the safety of the product is greatly improved, the usable dosage can be increased, and the clinical effect of the product is remarkably improved according to the dose-effect relationship.

One is as follows: through long-term and massive research and physical and clinical practice, the inventor invents the top leaf of the sparrow tea in a special period and quickly extracts and processes fresh tender leaves within 12 hours, thereby avoiding the toxicity increase caused by the hydrolysis of the effective component of the myrosinase in the storage process of the leaves;

secondly, the traditional theory (or cognition) of an ironware is avoided when the polyphenol compounds are extracted and separated, instead, a cast iron pot and pure water extraction are adopted to carry out production place processing, so that the effect that free (or hydrolyzed) hydroquinone is oxidized, polymerized and formed into a water-insoluble polymer under the catalysis of iron ions is achieved, the removal is easy, and the safety and low toxicity of the product are ensured;

thirdly, the technical scheme has the advantages of economy, practicability, easy industrialization and high product safety.

The special processing method, the proper dosage and the corresponding indications can better solve the problems of high uric acid and complications thereof, particularly high uric acid renal failure which seriously threatens life, and a plurality of life-threatening patients caused by high uric acid renal calculus have been successfully rescued according to the technical scheme.

The inventor of the technical scheme deeply researches the places where uric acid in a human body is generated, metabolized, transported, excreted and deposited to become stones and influences the kidney of the human body and the blood at the far end to flow smoothly, and the stones deposited to be enlarged stimulate the surrounding tissues to generate inflammation and initiate pathological changes. This is the root cause of the harm of high uric acid. In addition, the human body is a very complex and organic whole, and the prevention, treatment and rehabilitation of hyperuricemia and complications thereof must be a comprehensive integral treatment measure with multiple target points and multiple links, so that the effect can be achieved. And safe, stable and efficient products are the key. Therefore, according to the research on the safety of the tea leaves of the sparrow mouths, the inventor finds that the main toxic substances of the tea leaves are hydroquinone and parts (such as alkaloid and terpenoid) with high fat solubility, and the effective substances with better effect on reducing uric acid and complications thereof are caffeoyl arbutin effective parts with good water solubility, particularly, the inventor finds that the tea leaves are picked and processed in time when the content of the caffeoyl arbutin in tender leaves in 4-8 months is high, and the accompanying toxic substances are relatively more accumulated, so that the invention needs to adopt a proper extraction process to extract useful substances as far as possible and remove the harmful substances as far as possible, which is the reason that the technical scheme selects a green and environment-friendly process route for the whole-process pure water extraction in iron making equipment, and has the technical innovation points that: 1. the material is characterized in that tender leaves newly produced on the tip in months of 4-8 are selected, and the 6' -caffeoyl arbutin content is highest; 2. in the processing process, an ironware is used for promoting the oxidation and polymerization of hydroquinone toxic substances and the like, and then the hydroquinone toxic substances and the like are removed; 3. low-temperature vacuum drying is adopted to prevent oxidation and color deepening; 4. only water is used as the only solvent in the whole process, the cost is low, and the method is absolutely green and environment-friendly. The process has practical value suitable for producing area and further realizes the aim of the invention.

After obtaining the high-content 6' -O-caffeoyl arbutin extract with the content of more than 85 percent and the effective part with the hydroquinone (hydroquinone) <0.08 percent (namely the Quezui tea extract for treating diseases related to hyperuricemia, QZC-1060 for short), through years of clinical practice and exploring the relationship between the amount and the effect, the better dose (0.5-8g) for carrying out graded taking of different doses by patients is found, and the therapeutic dose is a higher dose, and various characteristics of the dose need to be grasped to obtain safe and effective results, which is also the place where the technical scheme has unexpected innovation and clinical effect.

Among the diseases caused by high uric acid, the most serious disease to life is end-stage complication renal failure, which is the disease difficult to treat so far, and most patients can only adopt dialysis to maintain low-quality survival. When a suitable source of transplanted kidney is found for kidney transplantation, it is necessary to administer an anti-rejection drug (cyclosporin or the like) which is expensive and has a large side effect for the whole life. In summary, renal failure is a medically refractory disease. The product provided by the technical scheme has unexpected treatment effect. The method adopts 2g-5g/d of 6' -O-caffeoyl arbutin with large dose, is taken for 4-6 times, drinks more water as much as possible, and can clean kidney, discharge kidney stone and recover kidney function until the patient is cured gradually (about 1 month) under the condition that the patient still has certain urine (100 ml/d). This is an unproductive result.

The inventors have found in the clinic a characterization of the product to function effectively: urine becomes smelly (strong ammonia smell) and then turns into ink color, and then the condition gradually improves. Among them, there is a possibility that urinary calculus may be scratched by surrounding tissues of urinary calculus and kidney calculus in the process of dissolving urinary calculus and kidney calculus gradually, which are the characteristics of effective function of the product.

In conclusion, the technical scheme provides an integrated and innovative treatment product for hyperuricemia and related diseases and a curative effect representation. It also provides a promising product for the medical problem of renal failure.

After a safe, effective and controllable effective part is obtained by using a determined integrated innovative process, according to deep and unique understanding of hyperuricemia and complications thereof, the inventor determines different using dosages and methods according to research, the product can be used for preventing, treating and recovering hyperuricemia and complications thereof in the whole process, can better embody the great health value of QZC-1060 and better provide a perfect and complete solution for patients with hyperuricemia and complications thereof: the dosage of the medicine for preventing the occurrence of hyperuricemia and the further deterioration of hyperuricemia patients is 0.5g-2 g/day; the dosage of the composition is 1.0g to 3g per day for patients with uric acid value of 480 mu mol/L to 580 mu mol/L and patients with gout; the dosage of the medicine is 1.5g to 4g per day for patients with uric acid value higher than 580 mu mol/L and patients with gout and uric acid calculus; the dosage of the composition is 2.0g-8 g/day for patients with uric acid value higher than 580 mu mol/L and patients with gout, uric acid stones and renal failure.

The preparation is selected, the oral preparation is convenient to use and low in cost, is suitable for prevention, treatment and rehabilitation, and is a basic product. For critical illness, injection, even intravenous injection, is selected according to special needs.

Selecting different raw materials, wherein the content of effective components is different, such as 5-7 months, 3-5 new leaves, and fresh processing, so that the extract has high 6' -O-caffeoyl arbutin content; conversely, if the leaf quality is poor, the content of the extract is small. The method is an innovative invention achievement after the technical scheme is explored in long-term hard practice.

The tea extract of the sparrow beak is used for preparing the medicine for treating the hyperuricemia and the complication, has multiple target points and multiple links, and has the action mechanisms of resisting inflammation, inhibiting bacteria, resisting oxidation and reducing the uric acid when acting on the hyperuricemia and the complication.

The inventor finds the effective functional characterization of the product in clinic: urine becomes smelly (strong ammonia smell) and then turns into ink color, and then the condition gradually improves. Among them, there is a possibility that urinary calculus may be scratched by surrounding tissues of urinary calculus and kidney calculus in the process of dissolving urinary calculus and kidney calculus gradually, which are the characteristics of effective function of the product.

Compared with the prior art, the invention has the beneficial effects that:

1. the technical scheme of the invention clearly limits the pretreatment process conditions of the tea leaves at the top of the tea leaves at a specific time period, and the tea leaves at the top of the tea leaves at a specific time period need to be quickly extracted within 12 hours, so that the toxicity is prevented from being increased due to the hydrolysis of the active ingredient of the uridine during the storage of the fresh leaves; if the extraction can not be completed within 12 hours, the water-removing and drying treatment is needed so as to be convenient for extraction after storage, and the generation of toxic substances by hydrolyzing the wujin glycoside can be effectively avoided.

2. The invention changes the prior art that the traditional theory or cognition of ironware is avoided during the extraction and separation of polyphenol compounds, namely the technical bias is overcome, the method adopts the processing of the nearby raw material producing area and uses an iron extraction container to realize the effect that free (or hydrolyzed) hydroquinone is oxidized, polymerized and formed into a water-insoluble polymer under the catalysis of iron ions, thereby being easy to remove and ensuring the safety and low toxicity of the product.

3. The invention also has the advantages of economy, practicability, easy industrialization and high product safety.

4. The special processing method, the proper dosage and the corresponding indications of the invention can better solve the problems of high uric acid and complications thereof, especially the high uric acid renal failure seriously threatening life, and a plurality of life-threatening patients caused by the high uric acid renal calculus can be successfully rescued according to the technical proposal.

The invention deeply researches the generation, metabolism, transportation, excretion and deposition of uric acid in a human body, and deposits large stones in the places which influence the kidney of the human body and have poor blood flow at the far end, so as to stimulate surrounding tissues to generate inflammation and initiate pathological changes. This is the root cause of the harm of high uric acid. In addition, the human body is a very complex and organic whole, and the prevention, treatment and rehabilitation of hyperuricemia and complications thereof must be a comprehensive integral treatment measure with multiple target points and multiple links, so that the effect can be achieved. And safe, stable and efficient products are the key.

5. According to the research on the safety of the tea leaves of the sparrow mouths, the main toxic substances of the tea leaves of the invention are hydroquinone and parts (such as alkaloid and terpenoid) with high fat solubility, and the effective substances with better effect on reducing uric acid and complications thereof are 6 '-caffeoyl arbutin effective parts with good water solubility, especially the tea leaves of 4-8 months are picked and processed in time when the 6' -caffeoyl arbutin content is high, and the accompanying toxic substances are relatively more gathered, so that the invention needs to adopt a proper extraction process to extract useful substances as far as possible and to extract harmful substances as far as possible, which is the reason that the technical scheme selects a green environment-friendly process route for whole-process pure water extraction in iron making equipment.

6. The raw materials of the invention are the tender leaves newly obtained from the apex in the month of 4-8, and the 6' -caffeoyl arbutin content of the tender leaves is highest; low-temperature vacuum drying is adopted to prevent oxidation and color deepening; only water is used as the only solvent in the whole process, the cost is low, and the method is absolutely green and environment-friendly. Organic solvent is not used, so that the harm to human bodies caused by the residual organic solvent is avoided.

7. The 6' -O-caffeoyl arbutin (effective part) of the invention is more than 85 percent, and hydroquinone (hydroquinone) is less than 0.08 percent, the preparation has high dose-efficiency ratio, and unexpected clinical effect is obtained. Not only for diseases caused by high uric acid, but also for renal failure which is the most serious to life and is the end-stage complication, the renal failure is the disease which is difficult to treat so far, and a relatively ideal treatment effect is achieved. The safe and large-dose 6' -O-caffeoyl arbutin is taken, so that the effects of cleaning the kidney, discharging kidney stones and recovering the kidney function can be achieved, and the healing effect can be achieved in about 1 month.

Drawings

FIG. 1 is an HPLC chromatogram of a sparrow tea and hydroquinone control; wherein 1-A is an HPLC chromatogram of the sparrow tea; 1-B is an HPLC chromatogram of a hydroquinone reference substance;

FIG. 2 is an HPLC chromatogram of 6' -O-caffeoyl arbutin, caffeic acid and oujin glycoside A; from left to right, the first chromatographic peak is caffeic acid; the second chromatographic peak is 6' -O-caffeoyl arbutin; the third chromatographic peak is the ouabain A;

FIG. 3 is an HPLC chromatogram of the Quezui tea extract of the present invention; from left to right, the first chromatographic peak is 6' -O-caffeoyl arbutin; the second chromatographic peak is ouabain A.

Detailed Description

The present invention will be described in further detail with reference to examples.

It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples do not specify particular techniques or conditions, and are performed according to the techniques or conditions described in the literature in the art or according to the product specifications. The materials or equipment used are not indicated by manufacturers, and all are conventional products available by purchase.

The percentage numbers are percentages by mass unless otherwise indicated.

Example 1

The sparrow tea extract for treating hyperuricemia and related diseases is characterized in that the mass content of 6' -O-caffeoyl arbutin in the sparrow tea extract is more than 85%, and the mass content of hydroquinone is less than 0.08%.

Example 2

The sparrow tea extract for treating hyperuricemia and related diseases is characterized in that the mass content of 6' -O-caffeoyl arbutin in the sparrow tea extract is more than 85%, and the mass content of hydroquinone is less than 0.08%.

Also contains 0.5 percent of phenolic compounds by mass, wherein the phenolic compounds comprise the ouabain A.

Example 3

The sparrow tea extract for treating hyperuricemia and related diseases is characterized in that the mass content of 6' -O-caffeoyl arbutin in the sparrow tea extract is more than 85%, and the mass content of hydroquinone is less than 0.08%.

The tea drink also contains 15% of phenolic compounds by mass, wherein the phenolic compounds comprise the ouabain A, caffeic acid and arbutin (the mass ratio of the three is 5: 1: 2).

Example 4

The sparrow tea extract for treating hyperuricemia and related diseases is characterized in that the mass content of 6-O-caffeoyl arbutin in the sparrow tea extract is more than 85%, and the mass content of hydroquinone is less than 0.08%.

Also contains 10% of phenolic compounds, wherein the phenolic compounds comprise caffeic acid and arbutin (the mass ratio is 1: 9).

Example 5

A preparation method of a sparrow tea extract for treating hyperuricemia and related diseases comprises the following steps:

step (1), selecting raw materials: selecting new young leaves of the maidenhair tea in the months of 4-8, and cleaning the fresh leaves with tap water after picking;

step (2), extraction: extracting the raw materials treated in the step (1) in an iron container; the time from picking to extraction is not more than 12 h;

the specific extraction method comprises the following steps: adding drinking water to cover the raw materials, boiling with strong fire for 30min (based on the boiling of the leaves), cooling to 40 deg.C, further crushing the leaves, cooling to 30 deg.C, filtering, cooling the filtrate to 20 deg.C, stirring for 30min, stopping stirring when white substance appears, and standing in dark for crystallization;

and (3) post-processing: removing supernatant of the crystallization solution under dark and heat-proof conditions, filtering or centrifuging, collecting crystals, washing the crystals with 4 deg.C water, and vacuum drying the crystals to obtain the extract of Focus Caraganae.

Example 6

A preparation method of a sparrow tea extract for treating hyperuricemia and related diseases comprises the following steps:

step (1), selecting raw materials: selecting new young leaves of the maidenhair tea in the months of 4-8, and cleaning the fresh leaves with tap water after picking;

step (2), extraction: extracting the raw materials treated in the step (1) in an iron container; the time from picking to extraction is not more than 12 h;

the specific extraction method comprises the following steps: adding drinking water to cover the raw materials, boiling with strong fire for 30min (based on the boiling of the leaves), cooling to 40 deg.C, further crushing the leaves, cooling to 30 deg.C, filtering, cooling the filtrate to 20 deg.C, stirring for 10min, stopping stirring when white substance appears, and standing in dark for crystallization;

and (3) post-processing: removing supernatant of the crystallization solution under dark and heat-proof conditions, filtering or centrifuging, collecting crystals, washing the crystals with 4 deg.C water, and vacuum drying the crystals to obtain the extract of Focus Caraganae.

If the tea cannot be extracted in time within 12 hours, the picked tea is firstly subjected to enzyme deactivation treatment, then dried into dry products at the temperature of 55 ℃, and then crushed or cut into coarse powder of 5 meshes to be directly used as an extracted medicinal material.

Further comprises a refining step; the refining step is 1 time;

adding water with the mass 2 times of that of the crystal into the washed crystal under a dark inert gas, heating and refluxing on a steam bath until the crystal is completely dissolved, then cooling to 75 ℃, then adding activated carbon for refluxing and decoloring, filtering, naturally cooling the filtrate for crystallization, filtering, washing the crystal with water with the temperature of 5 ℃, and then drying in vacuum to obtain the refined product of the tea extract.

The mass ratio of the activated carbon to the washed crystal is 50: 1000, parts by weight; the specific method for vacuum drying comprises the following steps: vacuum drying at 65 deg.C under 100mmHg for 14 hr.

Example 7

A preparation method of a sparrow tea extract for treating hyperuricemia and related diseases comprises the following steps:

step (1), selecting raw materials: selecting new young leaves of the maidenhair tea in the months of 4-8, and cleaning the fresh leaves with tap water after picking;

step (2), extraction: extracting the raw materials treated in the step (1) in an iron container; the time from picking to extraction is not more than 12 h;

the specific extraction method comprises the following steps: adding drinking water to cover the raw materials, boiling with strong fire for 40min (until the leaves are boiled), cooling to 60 deg.C, further mincing the leaves, cooling to 50 deg.C, filtering, cooling the filtrate to 40 deg.C, stirring for 35min, stopping stirring when white substance appears, and standing in dark for crystallization;

and (3) post-processing: removing supernatant of the crystallization solution under dark and heat-proof conditions, filtering or centrifuging, collecting crystals, washing the crystals with water of 7 deg.C, and vacuum drying the crystals to obtain the extract of Focus Caraganae.

If the tea cannot be extracted in time within 12 hours, the picked tea is firstly subjected to enzyme deactivation treatment, then dried into dry products at the temperature of 65 ℃, and then crushed or cut into 10-mesh coarse powder to be directly used as the extracted medicinal materials.

Further comprises a refining step; the refining step is 2 times;

adding water with the mass 3 times of that of the crystal into the washed crystal under a dark inert gas, heating and refluxing on a steam bath until the crystal is completely dissolved, then cooling to 85 ℃, then adding activated carbon for refluxing and decoloring, filtering, naturally cooling the filtrate for crystallization, filtering, washing the crystal with water with the temperature of 15 ℃, and then drying in vacuum to obtain the refined product of the hippophae petiolatus extract.

The mass ratio of the activated carbon to the washed crystal is 10: 1000, parts by weight; the specific method for vacuum drying comprises the following steps: vacuum drying at 55 deg.C under 80mmHg for 10 hr.

Example 8

A preparation method of a sparrow tea extract for treating hyperuricemia and related diseases comprises the following steps:

step (1), selecting raw materials: selecting new young leaves of the maidenhair tea in the months of 4-8, and cleaning the fresh leaves with tap water after picking;

step (2), extraction: extracting the raw materials treated in the step (1) in an iron container; the time from picking to extraction is not more than 12 h;

the specific extraction method comprises the following steps: adding drinking water to cover the raw materials, boiling with strong fire for 35min (based on the boiling of the leaves), cooling to 50 deg.C, further rubbing the leaves, cooling to 40 deg.C, filtering, cooling the filtrate to 30 deg.C, stirring for 35min, stopping stirring when white substance appears, and standing in dark for crystallization;

and (3) post-processing: removing supernatant of the crystallization solution under dark and heat-proof conditions, filtering or centrifuging, collecting crystals, washing the crystals with 6 deg.C water, and vacuum drying the crystals to obtain the extract of Focus Caraganae.

If the tea cannot be extracted in time within 12 hours, the picked tea is firstly subjected to enzyme deactivation treatment, then dried into dry products at the temperature of 60 ℃, and then crushed or cut into coarse powder of 8 meshes to be directly used as an extracted medicinal material.

Further comprises a refining step; the refining step is 1 or more times;

adding water with the mass 2.5 times of that of the crystal into the washed crystal under the dark inert gas, heating and refluxing on a steam bath until the crystal is completely dissolved, then cooling to 80 ℃, then adding activated carbon for refluxing and decoloring, filtering, naturally cooling the filtrate for crystallization, filtering, washing the crystal with water at 10 ℃, and then drying in vacuum to obtain the refined product of the tea extract.

The mass ratio of the activated carbon to the washed crystal is 30: 1000, parts by weight; the specific method for vacuum drying comprises the following steps: vacuum drying is carried out for 12h at 60 ℃ and 90 mmHg.

Example 9

A preparation method of a sparrow tea extract for treating hyperuricemia and related diseases comprises the following steps:

selecting new tender leaves of the maidenhair tea in 4-8 months, cleaning the fresh leaves with tap water after picking, and performing enzyme deactivation and dry product treatment at low temperature of less than 60 +/-5 ℃ as soon as possible if the fresh leaves cannot be extracted in 12 hours in time. Pulverizing (or cutting) into 5-10 mesh coarse powder, and directly extracting.

Step (2), putting 40kg of the processed medicinal materials into an iron pan, adding drinking water until the leaves are covered (8 times of water is added), boiling with strong fire until the leaves are boiled to be well, cooling to 50 +/-10 ℃, further rubbing the leaves, continuously cooling to 40 +/-10 ℃, filtering with gauze to obtain red filtrate, cooling to 30 +/-10 ℃, continuously starting slow stirring, stopping stirring when white substances appear, and standing in a dark place for crystallization;

and (3) carefully removing the supernatant of the crystallization solution in the dark and in the absence of heat, filtering, drying by filtration, washing the crystals once by 200ml of pure water with the temperature of 5 ℃, filtering, and drying in the dark at low temperature or in the shade. Or centrifugal filtering the precipitate, spin-drying, collecting filter cake, and cleaning to obtain the final product. 0.9-1.2 kg of white-like product (calculated on dry product) is obtained. The yield (relative to fresh medicinal materials) is 2.25 to 3 percent.

Step (4), when the purity of 6' -O-caffeoyl arbutin is more than 95%, under the condition of keeping out of the sun and charging nitrogen, taking 1kg of the sample into a 5L round-bottom flask, adding 2-3 times (2-3L) of distilled water, heating, refluxing and dissolving on a steam bath, slightly cooling to 80 +/-5 ℃, adding 10-50g of activated carbon for decoloration and refluxing for 10min, filtering, removing the activated carbon, collecting filtrate, naturally cooling the filtrate for crystallization, filtering, and washing the crystal with 10 +/-5 ℃ of distilled water;

and (5) taking out the wet extract, putting the wet extract into a vacuum drying oven, and performing vacuum drying for 12 +/-2 hours at the temperature of 60 +/-5 ℃ under the mmHg. The qualified finished product (QZC-1060) with the water content of less than 15 percent and the content of the active ingredients of 6' -O-caffeoyl arbutin of 96.0 percent to 99.5 percent is obtained, the yield of the step is more than 85 percent (calculated on dry products). This procedure can be repeated to obtain a purer sample.

And (6) packaging the qualified finished products into 1 kg/bag by using food-grade aluminum plastic bags with automatic sealing, and storing in a dry and light-proof environment. Can be used for preparing products for treating diseases caused by high uric acid. The name of the extract product is QZC-1060 for short.

Example 10

Weighing 200g of de-enzymed and dried sparrow mouth tea leaves, crushing the tea leaves into 10-mesh coarse powder, adding the coarse powder into a 5L iron pot, adding 20 times (4000ml) of purified water, heating and boiling for 0.5h, cooling the mixture to 60 ℃ by using cooling water, and filtering the mixture by using quick (coarse) filter cloth to obtain red filtrate; cooling to 40 deg.C, slowly stirring for 30min, stopping stirring when white substance appears, standing in dark for crystallization, and standing for 24 hr.

Carefully removing the supernatant of the crystallization solution in the dark and in the dark, filtering, draining, washing and crystallizing once with 20ml of pure water at 4-10 ℃, filtering, drying in the dark at low temperature or in the shade to obtain 4.1g of a white-like product finished product, namely the extract (called sparrow mouth tea extract for short) with high content of the active ingredients of 6' -O-caffeoyl arbutin. The detection shows that the content of 6' -O-caffeoyl arbutin (aurantiamarin) is 85.6 percent, the content of caffeic acid is 5.76 percent, the content of aurantiamarin A is 0.12 percent, the content of hydroquinone (hydroquinone) is 0.075 percent, and the yield is 2.0 percent. And packaging the dried product with a food-grade aluminum plastic bag with automatic sealing, and storing the packaged product in a dry and light-proof environment for later use.

Example 11

Selecting young leaves of the newly-released sparrow mouth tea picked in 7-8 months, cleaning the fresh leaves with tap water after picking, putting 40kg of processed tea leaves into an iron pot, adding drinking water with 6 times of the raw materials, immersing the raw materials into the water, and boiling for 40min with strong fire until the tea leaves are boiled to be well done. Cooling to 50 deg.C, further mincing the leaves, cooling to 40 deg.C, and filtering with gauze to obtain red filtrate; and cooling to 35 deg.C, stirring slowly for 40min, stopping stirring when white substance appears, standing in dark for crystallization, and standing for 24 hr. Carefully removing the supernatant of the crystallization solution in the dark and in the dark, filtering and draining, washing the crystals once with 200ml of pure water at 4-10 ℃, draining the water with a centrifuge, collecting filter cakes, and washing the crystals, wherein the crystals are the extract with high content of the 6' -O-caffeoyl arbutin effective components. Taking out the crystal, placing into a vacuum drying oven, and vacuum drying at 60 + -5 deg.C under 100mmHg for 14 hr to obtain dry extract. Wherein the content of the active ingredients of the 6' -O-caffeoyl arbutin is 0.96 kg of dry product with the content of 90.2 percent, the content of the aureoidin is 90.2 percent, the content of the aureoidin A is 6.80 percent, the content of hydroquinone (hydroquinone) is 0.067 percent, and the yield (relative to the raw material) is 2.4 percent. And packaging the dried product with a food-grade aluminum plastic bag with automatic sealing, and storing the packaged product in a dry and light-proof environment for later use.

Example 12

Selecting 3-5 leaves of the newly-grown young leaves of the tea leaves of the sparrow mouths picked in 4-5 months, crushing the fresh leaves into coarse powder of 5 meshes within 5 hours after picking, and directly using the coarse powder as an extraction raw material. Putting 80kg of coarse powder raw materials into an iron interlayer extraction tank, adding 12 times of drinking water into the extraction tank, and immersing the raw materials into the water. Introducing steam into the interlayer of the extraction tank, heating and boiling for 35min until the leaves are boiled to a state of being thoroughly cooked. Cooling the extraction tank to about 50 deg.C with cooling water in the interlayer, further mincing the leaves, cooling to 45 deg.C, and filtering with gauze to obtain red filtrate. And continuously cooling the filtrate to 35 ℃, starting to stir slowly for 35min, stopping stirring when white substances appear, and standing in a dark place for crystallization for 24 h. Carefully removing the supernatant of the crystallization solution in the dark and in the dark, filtering, draining, washing the crystals once with 400ml of pure water at 4-10 ℃, filtering, drying in the dark and air-drying at low temperature or drying in the shade. Collecting food, cleaning and crystallizing to obtain the extract with high content of 6' -O-caffeoyl arbutin effective component. Taking out the crystal product, placing the crystal product into a vacuum drying oven, and carrying out vacuum drying for 10h at the temperature of 60 +/-5 ℃ under the condition of 80mmHg to obtain 2.55 kg of a white extract dry product, wherein the content of the aureoidin is 95.5 percent, the content of the aureoidin A is 0.72 percent, the content of hydroquinone (hydroquinone) is 0.059 percent, and the yield (relative fresh raw material) is 3.18 percent. And packaging the dried product with a food-grade aluminum plastic bag with automatic sealing, and storing the packaged product in a dry and light-proof environment for later use.

Example 13

Selecting 1kg of the dried crystal obtained in example 12 (wherein the content of the aurantiamarin is 95.5%, the content of the aurantiamarin A is 0.72%, and the content of the hydroquinone (hydroquinone) is 0.059%), adding 2-3 times (2-3L) of distilled water into a 5L round-bottomed flask under dark condition and nitrogen filling, heating and refluxing on a steam bath under dark condition until the crystal is completely dissolved, slightly cooling to 80 +/-5 ℃, adding 25g of activated carbon, decoloring and refluxing for 10min, filtering, removing the activated carbon, collecting filtrate, naturally cooling the filtrate for crystallization, filtering, washing the crystal with 50ml of 10 ℃ distilled water, placing the refined crystal into a vacuum drying oven, and carrying out vacuum drying for 13h at 60 +/-5 ℃ under 100 mmHg. 0.86kg of white product dry product is obtained, wherein the content of the uridine is 99.2 percent, the content of the uridine A is 0.19 percent, the content of the hydroquinone (hydroquinone) is 0.020 percent, and the yield is 86 percent. The process is a refining process, and a refined product with higher purity can be obtained by repeating the process. The refined product is packaged into 0.5 kg/bag by using a food-grade plastic bag with automatic sealing, and is stored in a dry and light-proof environment for preparing the medicine for treating the related diseases caused by hyperuricemia.

Through experimental research, the inventor confirms that the safe, effective and controllable effective part (QZC-1060) of the griffonia simplicifolia extract can be obtained under the integrated innovative process of the invention. According to the deep and unique understanding of hyperuricemia and the complication thereof, the inventor determines different using dosages and using methods of the preparation through animal and clinical experimental researches. The product of the invention can be used for preventing, treating and recovering hyperuricemia and complications thereof in the whole process, can better embody the great health value of QZC-1060 and better provide a perfect and complete solution for patients suffering from hyperuricemia and complications thereof.

Clinically, the dosage (by the content of active ingredients) of a patient is 2-6 g/day, and 1-3 times/day is suitable. For preventing the occurrence of hyperuricemia and patients with further worsening of hyperuricemia, the dosage is 2-6 g/day and 1-3 times/day. For patients with uric acid value higher than 480 mu mol/L and patients with gout, the dosage is 2-3 g/day and 1-3 times/day; for patients with uric acid value higher than 580 mu mol/L and patients with gout and uric acid stones, the dosage is 2-4 g/day and 2-3 times/day; for patients with uric acid value higher than 580 mu mol/L and patients with gout, uric acid stones and renal failure, the dosage is 2-5 g/day and 3 times/day.

The invention selects the oral preparation, has convenient use and low cost, is suitable for prevention, treatment and rehabilitation, and is a basic product. For critical and serious diseases, the injection is selected according to special requirements, and the intravenous administration injection is also safe.

Example 14

The preparation of the standard extract capsule of the sparrow tea comprises the following components by 1000 granules:

50g of sparrow tea extract (QZC-1060), 16g of dextrin, 9g of microcrystalline cellulose, 7g of sodium methyl starch and 1g of magnesium stearate. And the preparation method comprises the following steps: weighing QZC-1060, dextrin, microcrystalline cellulose and sodium carboxymethyl starch according to the prescription amount, putting the mixture into a wet granulator, uniformly mixing (mixing for 10min +/-1 min), and adding the mixture with the volume concentration of 50% ethanol, and setting granulation process parameters (stirring speed of 700r min)^-1The cutting speed is 1000 r.min^-1Granulating for 5min), drying, controlling water content of granule at 5%, grading (20 mesh sieve), adding magnesium stearate into the granule, mixing, and making into No. 2 capsule.

Example 15

The preparation of the standard extract capsule of the sparrow tea comprises the following components by 1000 granules:

110g of the tea extract of the bird's beak (QZC-1060), 28g of dextrin, 10g of microcrystalline cellulose, 5g of plantain seed gum, 10g of konjac glucomannan, 10g of sodium methyl starch and 1g of magnesium stearate. And the preparation method comprises the following steps: weighing QZC-1060, dextrin, microcrystalline cellulose, plantain seed gum, konjac polysaccharide and sodium carboxymethyl starch according to the prescription amount, placing the materials in a wet granulator, mixing uniformly (mixing for 10min +/-1 min), adding ethanol with the volume concentration of 50 percent, and setting the granulation process parameters (the stirring speed is 700 r.min)^-1The cutting speed is 1000 r.min^-1Granulating for 5min), drying, controlling water content of the granules to be 5%, grading (20-mesh screen), adding appropriate amount of magnesium stearate into the granules, mixing, and encapsulating into No. 1 capsule.

Example 16

The preparation of the standard extract tablet of the sparrow tea comprises the following components in a formula of 1000 tablets:

200g of sparrow tea extract (QZC-1060), 41g of dextrin, 20g of microcrystalline cellulose, 5g of plantain seed gum, 10g of konjac polysaccharide, 18g of sodium methyl starch and 1g of magnesium stearate. And the preparation method comprises the following steps: weighing QZC-1060, dextrin, microcrystalline cellulose, plantain seed gum, konjac polysaccharide and sodium carboxymethyl starch according to the prescription amount, placing the materials in a wet granulator, mixing uniformly (mixing for 10min +/-1 min), adding ethanol with the volume concentration of 50 percent, and setting the granulation process parameters (the stirring speed is 700 r.min)^-1The cutting speed is 1000 r.min^-1Granulating for 5min), drying, controlling water content of the granules to be 5%, grading (20-mesh screen), adding appropriate amount of magnesium stearate into the granules, mixing, tabletting with a tabletting machine according to 1000 tablets, and packaging.

Example 17

The preparation of the standard extract tablet of the sparrow mouth tea comprises the following components in parts by weight of 10000 granules:

6000g of sparrow tea extract (QZC-1060), 600g of dextrin, 280g of microcrystalline cellulose, 180g of plantain seed gum, 200g of konjac polysaccharide, 140g of sodium methyl starch and 10g of magnesium stearate. And the preparation method comprises the following steps: weighing QZC-1060, dextrin, microcrystalline cellulose, plantain seed gum, konjac polysaccharide and sodium carboxymethyl starch according to the prescription amount, placing the materials in a wet granulator, mixing uniformly (mixing for 10min +/-1 min), adding ethanol with the volume concentration of 50 percent, and setting the granulation process parameters (the stirring speed is 700 r.min)^-1The cutting speed is 1000 r.min^-1Granulating for 5min), drying, controlling water content of the granules to be 5%, grading (20-mesh screen), adding appropriate amount of magnesium stearate into the granules, mixing, tabletting according to 10000 tablets with a tabletting machine, and packaging to obtain the final product. Each tablet contains 600mg of the extract of the tea of Quezui, and the product with the enlarged prescription is selected clinically.

Example 18

The preparation of the standard extract granules of the sparrow mouth tea comprises the following components in 1000 bags:

400g of the tea extract of the bird's beak (QZC-1060), 53g of dextrin, 31g of microcrystalline cellulose, 12g of plantain seed gum, 31g of konjac glucomannan, 21g of sodium methyl starch and 1.1g of magnesium stearate. And the preparation method comprises the following steps: weighing QZC-1060, dextrin, microcrystalline cellulose, plantain seed gum, konjac polysaccharide and sodium carboxymethyl starch according to the prescription amount, placing the materials in a wet granulator, mixing uniformly (mixing for 10min +/-1 min), adding ethanol with the volume concentration of 50 percent, and setting the granulation process parameters (the stirring speed is 700 r.min)^-1The cutting speed is 1000 r.min^-1Granulating for 5min), drying, controlling water content of granules at 5%, grading (20 mesh screen), packaging into packaging bags according to 1000 bags, and sealing.

Example 19

The standard extract oral liquid of the sparrow tea comprises the following components in 1000 bottles (10 ml/bottle): 170g of the tea extract (QZC-1060), 40.3g of plantain seed gum, 61g of konjac polysaccharide and 7g of stevioside, the raw materials are mixed, water is added to 10000ml of the prescription amount, the mixture is dissolved, and then 1000 bottles of the mixture are subpackaged and sterilized to obtain the tea.

Example 20

The standard sparrow tea extract freeze-dried powder injection comprises the following components in parts by weight of 1000 (2 ml/piece): 100g of standard extract of the tea of sparrow mouth (QZC-1060, wherein the content of the ouabain is more than 99 percent), 50g of mannitol and 2000ml of water for injection. The preparation process of the freeze-dried powder injection for injection comprises the following steps: under the condition of CGMP, taking the prescription amount of raw materials, adding water for injection to 1600ml, heating to 50 ℃, adding water for injection to the total preparation amount of 2000ml after dissolving, stirring uniformly, roughly filtering by 0.45 mu m, finely filtering by 0.2 mu m, filling 2ml of liquid medicine into a 5ml penicillin bottle, freezing and drying in vacuum to obtain the freeze-dried powder injection for injection with the specification of 100 mg.

Example 21

The standard sparrow tea extract freeze-dried powder injection comprises the following components in parts by weight of 1000 (2 ml/piece): 200g of standard extract of the tea of sparrow mouth (QZC-1060, wherein the content of the ouabain is more than 99 percent), 60g of mannitol and 2000ml of water for injection. The preparation process of the freeze-dried powder injection for injection comprises the following steps: under the condition of CGMP, taking the prescription amount of raw materials, adding water for injection to 1600ml, heating to 50 ℃, adding water for injection to the total preparation amount of 2000ml after dissolving, stirring uniformly, roughly filtering by 0.45 mu m, finely filtering by 0.2 mu m, filling 2ml of liquid medicine into a 5ml penicillin bottle, freezing and drying in vacuum to obtain the freeze-dried powder injection for injection with the specification of 200 mg.

Example 22

The standard sparrow tea extract freeze-dried powder injection comprises the following components in parts by weight of 1000 (2 ml/piece): 400g of standard extract of the tea of sparrow mouth (QZC-1060, wherein the content of the ouabain is more than 99 percent), 80g of mannitol and 2000ml of water for injection. The preparation process of the freeze-dried powder injection for injection comprises the following steps: under the condition of CGMP, taking the prescription amount of raw materials, adding water for injection to 1600ml, heating to 50 ℃, adding water for injection to the total preparation amount of 2000ml after dissolving, stirring uniformly, roughly filtering by 0.45 mu m, finely filtering by 0.2 mu m, filling 2ml of liquid medicine into a 5ml penicillin bottle, freezing and drying in vacuum to obtain the freeze-dried powder injection for injection with the specification of 400 mg.

The technical scheme is that the caffeoyl-rich arbutin effective part, the proper dosage and the proper preparation of the tea leaves of the sparrow mouths are integrated and innovated into an innovative scheme for effectively preventing, treating and recovering hyperuricemia and complications thereof, and the innovative scheme integrates the development of multiple disciplines.

The invention has the technical effect verification experiment:

drug effect screening experiment of Quezui tea extract QZC-1060 with different caffeoyl arbutin contents

1. Purpose of experiment

Taking a mouse hyperuricemia model induced by potassium oxonate as a drug effect screening platform, taking febuxostat and allopurinol as positive control drugs, carrying out dose conversion on each sample by using equimolar amount of caffeoyl arbutin, measuring blood uric acid, carrying out research on the uric acid reduction effect of the total extract of the tea leaves of the sparrow nozzle and the extract of the tea leaves of the sparrow nozzle with the caffeoyl arbutin content of (65%), (70%), (75%), (80%), 85%, 90%, 95% and 99.5%, and comparing the activity of the samples for reducing uric acid. (the extraction process is followed, and different herbs are selected to obtain different content of extract, which is not repeated here)

2. Experimental Material

2.1 Experimental animals: KM mice, male, 18-22 g, were provided by Schlekshirta laboratory animals Co., Ltd. (license number: SCXK (Hunan) 2009-0004) in Hunan, and were used for experiments after 3 days of adaptive feeding.

2.2 Experimental samples: 65 percent, 70 percent, 75 percent, 80 percent, 85 percent, 90 percent, 95 percent and 99.5 percent of the maidenhair tea extract; ninthly, a positive drug febuxostat (the content of the raw material drug is more than 98%); the drug allopurinol (Shanghai Xinyi pharmaceutical industry) positive for (R).

2.3 Experimental reagents: potassium Oxazinate (Sigma), 0.5% CMC-Na

3. Experimental methods

3.1 reagent preparation: firstly, mixing a test sample with positive control drugs of febuxostat and allopurinol: preparing and uniformly mixing with pure water; ② Potassium Oxonate: is prepared by 0.5 percent of CMC-Na and is mixed evenly before use.

3.1 grouping and administration

Because the screening samples are more, the screening work can be carried out by dividing the two batches of experiments, and the two batches of experiments are different in groups.

3.1.1 first batch: 160 male mice are randomly divided into a normal group, a model group, a (first) extract group high, medium and low dose group, a (second) extract group high, medium and low dose group, a (third) extract group high, medium and low dose group, a (fourth) extract group high, medium and low dose group, a positive drug febuxostat (crude drug with the content of more than 98 percent) group and a positive drug allopurinol group, and the total number of the mice is 16, and each group comprises 10 mice; normal group and model group are filled with normal saline, the administration groups are filled with corresponding drugs (the high, medium and low doses are respectively 5mg/kg, 10mg/kg and 20mg/kg) and 0.2ml/10g, the allopurinol group is filled with drugs at 30mg/kg, and the administration is carried out for 5 days.

3.1.2 second batch: male mice 160 were randomly divided into normal group, model group, total extract high, medium and low dose group, fifthly extract high, medium and low dose group; sixthly, the extract groups are high, medium and low dosage groups; seventhly, the extract groups comprise high, medium and low dose groups; the high, medium and low dosage groups of the eight extract; the positive drug febuxostat (crude drug with the content of more than 98%) group, the positive drug allopurinol group, 16 groups, 10 in each group; normal group and model group are filled with stomach physiological saline, the administration groups are filled with corresponding drugs (the high, medium and low doses are respectively 5mg/kg, 10mg/kg and 20mg/kg) and 0.2ml/10g, the allopurinol group is filled with stomach according to the dose of 30mg/kg, and the febuxostat group is filled with stomach according to the dose of 6mg/kg and is administered for 5 days.

3.2 establishment of hyperuricemia model and index detection

1h before the last administration, injecting oteracil potassium salt (450mg/kg) into the abdominal cavity except the normal group, administering 0.5% CMC-Na with equal dose to the normal group, taking blood from the retroorbital venous plexus of the mice after 1h after administration, standing for 1h at room temperature, centrifuging at 3500r/min for 15min to separate serum, and detecting the level of serum Uric Acid (UA) by using a full-automatic biochemical analyzer.

4. Results of the experiment

The experimental results are as follows: as can be seen from Table 3, uric acid in a hyperuricemia mouse model group caused by oteracil potassium is remarkably increased compared with a normal group, and the positive group allopurinol and febuxostat can remarkably reduce serum uric acid (P is less than 0.05); in the administration group, the high dose group of the extract group shows effective trend, and the low dose group of the 85 percent extract group has obviously reduced mouse serum uric acid (P is less than 0.05). Some of the results are shown in Table 3.

TABLE 3 Effect of different amounts of tea extract of Quezui on serum Uric Acid (UA) of hyperuricemic mice

Note: p <0.05, P < 0.01, compared to model group; compared with the normal group, # P <0.05, # P < 0.01

And (3) knotting: the study results showed that the 70% extract group showed an effective trend to the 85% extract group, which showed a significant reduction in serum uric acid in mice (P < 0.05). The content is increased, and the activity is still improved to a certain extent. In addition, the product has higher requirements on safety in consideration of the characteristics of long-term administration, so that the caffeoyl arbutin with the concentration of 85.0-99.5% has better safety and effectiveness in comprehensive consideration of effect-safety.

5. Discussion of the related Art

5.1 Potassium Oxonate is a uricase inhibitor, and the uricase activity in a mouse is inhibited, so that the uric acid decomposition is reduced, and the serum uric acid is rapidly increased to cause a hyperuricemia model. The model is an internationally recognized hyperuricemia animal model, the model is adopted in the experiment to screen the effective parts of the caffeoyl arbutin in the tea leaves of the sparrow mouth, the experimental result shows that the significance of uric acid in a model group is higher than that in a normal group, and the significance of uric acid in a positive drug can be reduced, so that the indication is that the hyperuricemia model established in the experiment is successful and can be used for drug effect evaluation.

Of the 5.28 samples tested, the 70% extract group showed an effective trend until the 85% extract group showed significant reduction of mouse serum uric acid (P < 0.05) in the low dose group, while the more pure 90%, 95%, 99.5% showed no significant uric acid reduction. The suggestion is that the uric acid reducing effect of the effective part of the caffeoyl arbutin is not increased along with the improvement of the purity of the caffeoyl arbutin. This may be related to the absorption properties of the creoso arbutin active fraction of Quezuela tea.

Second, the experiment of the influence of the extract of the tea broom cypress on the urine protein of the rat with chronic nephritis caused by adriamycin

1. Test article

Folium et cacumen Myristicae extract QZC-1060 (radix Linderae Glaucae extract with 95.5% of uridine); benazepril hydrochloride tablets, national standard character H20030514, 10 mg/tablet, manufactured by Beijing Nouhua pharmaceutical Co., Ltd., lot number X1774.

2. Primary reagent

Doxorubicin hydrochloride for injection (also called adriamycin hydrochloride), a national standard character H44024359, 10 mg/bottle, Shenzhen Wanle pharmaceutical industry Limited company, a batch number 0806E1, stored at 2-8 ℃, and prepared into 1mg/mL by a sodium chloride injection when in use; sodium chloride injection, 500 ml/bottle, Guizhou Tiandi pharmaceutical industry, Limited liability company, batch number A11102207; heparin sodium, 500 mg/count, beijing dingguo biotechnology ltd liability company, lot number 85810250; urine protein quantitative kit (CBB method), 96T/portion, Nanjing institute of bioengineering, lot number 20120316; SOD kit (xanthine oxidation), 96T/part, Nanjing institute of bioengineering, lot number 20120313; 96T/part of MDA kit (thiobarbituric acid method), Nanjing institute of bioengineering, lot number 20120310; NO kit (one-step method), 96T/part, Nanjing institute of bioengineering, lot number 20120802.

3. Animal(s) production

The SPF male SD rat is 6-8 weeks old and 200-250 g in weight, and is provided by Kunming medical college experimental animal center, and the production license is as follows: SCXK (Dian) 2005-: kunming City department of science and technology; raising the animals in an IVC animal laboratory, wherein the temperature is 20-25 ℃ (the daily temperature difference is less than or equal to 3 ℃), the humidity is 40-70%, and the illumination is 12 h: 12h light and shade are alternated, the illumination is 150-300 lx, the noise is less than or equal to 60dB, and the experimental animal use license is as follows: SYXK (yun) 2011-: kunming City department of science and technology; the PVC transparent plastic boxes are used for group culture, each box is less than or equal to 6, the mice are fed with compound feed every day, water is freely drunk, cages and padding are replaced according to the situation, the feed comes from the experimental animal center of Kunming medical college, and the production license is as follows: SCXK (Dian) 2011-: kunming City department of science and technology.

4. Test method

Male SD rats 40 were adaptively fed in the IVC animal room for 10 days, and 30 other rats were used for molding except for 10 normal controls, and each rat tail vein was injected with 0.1% doxorubicin hydrochloride solution at doses of 2, and 1mg/kg for 1 day, 7 days, and 14 days, respectively, and the normal controls were injected with physiological saline of equal volume. Urine is collected after 21 days of molding, and 24-hour urine protein is measured, and the result shows that the urine protein of most animals for 24 hours is generally increased, which indicates that the model is successful.

The 30 successfully molded rats are evenly divided into 3 groups according to the total amount of urine protein in 24 hours and the body weight: model control group 10, new lodine group 10, and que zui tea extract dose groups 10 each. Each group of animals was administered once daily by gavage at a dose of 56 days continuously with a volume of 10mL/kg.bw, and normal and model control groups were given 1% CMC-Na at equal volumes. Weigh 1 time per week and adjust the dose according to body weight.

5. Results of the experiment

The urinary protein levels were approximately uniform at 24h prior to dosing for each group and were significantly higher than the blank control group, indicating approximately equivalent kidney damage in each model group prior to dosing. During the administration period, the urinary protein level of the animal in the model control group is continuously increased and is obviously higher than that of the animal in the normal control group; the increase of urine protein caused by doxorubicin hydrochloride is reduced to different degrees by 56 days of continuous administration of each administration group, wherein the urine protein measurement results of 21 days after the administration of the positive control group, 49 days and 56 days after the administration of the QZC-1060 low dose group and the urine protein measurement results of 7 days, 28 days, 35 days, 42 days and 56 days after the administration of the high dose group are obviously lower than those of the model control group.

It is shown that QZC-1060 has therapeutic effect on kidney injury.

TABLE 4 Effect of 8 weeks of administration on model rat urine protein (+ -SD)

Compared with the control group: p is less than 0.05/0.01; compared to the model set: p < 0.05/0.01.

The table shows that the sparrow tea extract can obviously reduce the proteinuria of the adriamycin nephritis model rats after being continuously administered by gastric lavage for 8 weeks at the dose of 20mg/kg, and has obvious protective effect on the kidney injury and the kidney function.

Table 593 observation and comparison table for improving effect of peacock mouth tea tablet taken by gout patients

The total number of evaluators: 93 cases, 77 cases were valid, 16 cases were not obvious (invalid). Among the effective cases, 41 good cases account for 44.1%; the remissions were 36 cases, accounting for 38.7%; the number of the patients who were not obvious (ineffective) was 16, and the number of the patients accounted for 17.2%.

Total effective rate (%) + remission 44.1+38.7 ═ 82.8

Note: the samples used for the human clinical study were those from example 8, up to 10 ten thousand tablets, each containing 600mg of the extract.

Gastroemia acutifolia (QZC-1060) mice gavage acute toxicity test

1. Test materials

Test samples: QZC-1060 (wujin glycoside content 90.2%)

Experimental animals: experimental animal center of university of Kunming medical science, SPF class Kunming mouse, and the license number of experimental animal: SCXK (Dian.) 2015-0002.

2. Test method

Animal grouping: the mice of 60 groups, each group containing 10 mice and half of the mice, were divided into blank groups and 5 dose groups, i.e. 6000, 4000, 2000, 1000, 500 mg/kg.

Test samples are prepared into suspension with distilled water, animals in 5 dose groups are subjected to intragastric administration according to the equal volume of 20ml/kg. bw each time, the administration volume is calculated according to the body weight of each mouse, the administration dose (6000, 4000, 2000, 1000 and 500mg/kg) is achieved by two intragastric administrations at the interval of 7 hours, and the suspension of a test object is shaken uniformly before each intragastric administration.

The observation was continued for 14 days as usual.

The statistical method comprises the following steps: the death number of each group of animals was statistically analyzed by Bliss statistical method using SPSS12.0 software, and LD50 values were calculated.

3. Test results

The test results are shown in Table 6.

TABLE 6 acute toxicity test results of gavage of QZC-1060 mice

4. Conclusion

At toxic doses, the possible target organs that produce toxicity are the liver and nervous system. The toxicity to females may be greater than for males. The LD50 value is 5437.9mg/kg, the 95% confidence interval is 3790.1-15403mg/kg, the Quezui tea extract QZC-1060 is low-toxicity substance according to the acute toxicity classification of chemical toxicity identification technical code.

Four, typical cases

1. In a certain disease, 78 years old, retired workers in the geological team of Xiangfan city, Hubei, suffered from gout for more than 20 years, had stones on hands and feet, but were not large, and left kidney stones reached 2.2 cm, and were diagnosed as moderate hydronephrosis, and the Beijing hospital had no hospital bed. The sparrow mouth tea tablet of the invention is taken at 7 months in 2019, and 12 tablets (7200 mg/day) are taken every day till now.

Creatinine 227, urea 11.69, and uric acid 704 were examined in 2019, 8/31/month. The decrease of the muscular liver is detected to be 175 in 2019 in 10 months, urea is 7.09, uric acid 584 is used, gout is not painful, the walking can be carried out for more than five kilometers every day, and the daily health can be achieved. Creatinine 162, uric acid 543 and urea 11.44 are detected in 2020, 1 month and 4 days. Originally, the three upstairs and downstairs are difficult, and 25 kilograms of water can be freely lifted and bottled. Creatinine 158.8, urea 10.59 and uric acid 485 mu mol/L are checked for 21 days in 6 months in 20 years.

2. Qiu, male 64 years old, familial hyperuricemia, many people in the family have high uric acid, the uric acid of the person is detected to fluctuate in the range of 500-.

796 mu mol/L of 2020-11-26 blood uric acid examination can cause complications when the patient is careful about long-term hyperuricemia. The sparrow mouth tea tablets are taken at 2020-12-11 days, 3 tablets per day, one tablet in the morning, at noon and at night. 2020-12-27 the uric acid index of nearly half a month after the tea tablet is taken at home is reduced to 503, and the left sole feels a little. 383 pieces of uric acid were measured by oneself on 2021-1-9 days, 428 pieces of uric acid were measured by oneself on 1-16 days, and the uric acid is increased back, the estimated amount of tea pieces is small, 6 pieces/day and 2 pieces/3 times are added on 2021-17 days. 21-1-25 self-test uric acid 486 without pain and other uncomfortable feelings, self-test uric acid 356 in 2021-1-31 days, change from 2021-2-1 to 4 tablets per day, and self-test uric acid 368 in 2021-2-18 days. The effect is stable when 4 tea tablets are taken per day, the blood uric acid in 2021-3 months is in the range of 355-.

3. In Yang Jie, male age 65 years, the blood uric acid is high, 520umol/L, and the blood uric acid is reduced to 480 without pain after people pay attention to diet. Pain appeared after drinking beer seafood after going to Thailand in 2018, and after coming back, the medicine was treated with benzbromarone and a pain-relieving medicine. Pain occurs several times a year in the last two years. 2020-12-23 people start to take the tea tablet of the invention, 3 tablets are taken per day, one tablet is taken in the morning, at noon and evening, the tea tablet is taken for 1 month (21-1-23 follow-up), no food is prohibited and no disease is caused between 370 and 420 medicines for self-testing, and the feeling is effective. When the fish is played in spring festival, 3 times of chafing dish and 2 times of sour and hot fish are eaten, the tea slices are not taken for several days, the uric acid is increased to 570, the pain is avoided, the uric acid is eaten 6 slices/day and 2 slices/3 times, and the uric acid is reduced to 460 after the fish is eaten for several days. The number of the tablets per day is changed from 2021 to 3 months, and the uric acid is controlled to be about 400 mu mol/L.

The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.