RU2780451C1 - Composition based on medicinal materials of traditional chinese medicine for treatment of metabolic syndrome, its production method and use, and pharmaceutical drug based on it and its use - Google Patents

Abstract

FIELD: pharmaceutics.

SUBSTANCE: group of inventions relates to the pharmaceutical industry, namely to a composition for the treatment of metabolic syndrome. A composition based on medicinal materials of traditional Chinese medicine for the treatment of metabolic syndrome is proposed, where the composition based on medicinal materials of traditional Chinese medicine contains Coptidis Rhizoma, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Trichosanthis Fructus, Cinnamomi Cortex, and red yeast rice in the form of raw materials. At the same time, the composition contains 19.57 wt. % of Coptidis Rhizoma, 19.56 wt. % of Aurantii Fructus Immaturus, 19.57 wt. % of Pinelliae Rhizoma, 26.09 wt. % of Trichosanthis Fructus, 2.17 wt. % of Cinnamomi Cortex, and 13.04 wt. % of red yeast rice. The composition based on medicinal materials of traditional Chinese medicine for the treatment of metabolic syndrome is described, where the composition contains Coptidis Rhizoma, Aurantii Fructus Immaturus, red yeast rice, and optionally one or more components selected from a group consisting of Pinelliae Rhizoma, Trichosanthis Fructus, and Cinnamomi Cortex in the form of raw materials. In this case, the composition contains: 7.9-45 wt. % of Coptidis Rhizoma, 8.5-32.1 wt. % of Aurantii Fructus Immaturus, 0-33.8 wt. % of Pinelliae Rhizoma, 0-26.09 wt. % of Trichosanthis Fructus, 0-13.1 wt. % of Cinnamomi Cortex, and 13.04-57.2 wt. % of red yeast rice. A pharmaceutical composition for the treatment of metabolic syndrome contains the composition based on medicinal materials of traditional Chinese medicine and a pharmaceutically acceptable carrier. A method for the production of a composition for the treatment of metabolic syndrome from the above-described composition is proposed, where (a-1) Coptidis Rhizoma is extracted using an aqueous acid solution and filtered, concentrated, subjected to alkaline precipitation, salted, filtered, dried, and crushed to obtain an extract of the sum of alkaloids from Coptidis Rhizoma; (a-2) four medicinal materials, including red yeast rice, which makes up 90% of the amount required by the recipe, Aurantii Fructus Immaturus, Pinelliae Rhizoma, and Cinnamomi Cortex, are extracted using 70% ethanol and filtered; (a-3) Trichosanthis Fructus is extracted using 85% ethanol, the solution of Trichosanthis Fructus extract is combined with a solution of the extract of four medicinal materials of traditional Chinese medicine, ethanol is removed at reduced pressure, concentrated, dried, and crushed to obtain a mixed extract of five medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice, Cinnamomi Cortex, and Trichosanthis Fructus; and (a-4) the extract of the sum of alkaloids from Coptidis Rhizoma, the mixed extract of five medicinal materials of traditional Chinese medicine, and fine powder of red yeast rice, which makes up 10% of the amount required by the recipe, are mixed to a homogeneous state to obtain a composition based on medicinal materials of traditional Chinese medicine. A pharmaceutical drug for the treatment of metabolic syndrome containing a composition based on medicinal materials of traditional Chinese medicine is proposed, where the pharmaceutical drug provides injectable solutions, tablets, capsules, granules, and solutions for oral administration. The use of a composition based on medicinal materials of traditional Chinese medicine is proposed for the treatment of a disease selected from: a. metabolic syndrome; b. disorder of a blood glucose level and lipid metabolism at an early stage of the metabolic syndrome. The use of a composition based on medicinal materials of traditional Chinese medicine is proposed to provide one or more of following effects: lowering a blood glucose level, lowering a blood insulin level, improving glucose tolerance according to the result of an oral test, lowering a level of glycated hemoglobin (HbAlc), increasing an index of insulin sensitivity, improving a condition in insulin resistance, lowering a level of triglycerides in blood, lowering total cholesterol, and lowering blood pressure. The use of a pharmaceutical drug is proposed for the treatment of a disease selected from: a. metabolic syndrome; b. disorder of blood glucose level and lipid metabolism at an early stage of the metabolic syndrome. The use of a pharmaceutical drug is proposed to provide one or more of following effects: lowering a blood glucose level, lowering a blood insulin level, improving glucose tolerance according to the result of an oral test, lowering a level of glycated hemoglobin (HbAlc), increasing an index of insulin sensitivity, improving a condition in insulin resistance, lowering a level of blood triglycerides, lowering total cholesterol, and lowering blood pressure.

EFFECT: above -described compositions are effective for the treatment of metabolic syndromes, namely relatively to lowering a blood glucose level, lowering a blood insulin level, improving glucose tolerance according to the result of an oral test, lowering a level of glycated hemoglobin (HbAlc), increasing an index of insulin sensitivity, improving a condition in insulin resistance, lowering a level of blood triglycerides, lowering total cholesterol, and lowering blood pressure.

10 cl, 24 tbl, 12 ex

Description

FIELD OF TECHNOLOGY

The present invention relates to the field of medical technology, in particular, to a composition based on medicinal materials of traditional Chinese medicine, a method for its preparation, use, and pharmaceutical preparations based on it and their use.

BACKGROUND OF THE INVENTION

Metabolic syndrome is a clinical syndrome that has a serious impact on human health. The main symptoms of the metabolic syndrome include impaired glucose tolerance, arterial hypertension, hyperlipidemia, and central obesity. The common pathological basis of the metabolic syndrome is insulin resistance, and its clinical features include a constellation of various metabolic diseases. Currently, one-fourth to one-third of the world's adults suffer from metabolic syndrome. As the population ages and overweight/obesity spreads, the incidence/frequency of metabolic syndrome increases every year. Over the past 20 years, the incidence of metabolic syndrome in the US adult population has increased from 21% to 39%. In China, in the age group over 20 years, according to the results of diagnostics according to the WHO and NCEP-ATPIII standards, the incidence of metabolic syndrome was 14%-18% and 12%-21%, respectively. In patients with metabolic syndrome, the incidence of cardiovascular disease and the likelihood of stroke and death is 2-3 times higher than in patients without metabolic syndrome. The risk of developing type 2 diabetes in patients with metabolic syndrome is 5 times higher than in patients without metabolic syndrome. Among patients with metabolic syndrome, about 45% are patients with abnormal glucose metabolism, more than 80% are patients with abnormal blood lipid levels, and approximately 90% are patients with high blood pressure. To date, the control of metabolic syndrome in Western medicine mainly involves the combined use of a number of drugs, which are characterized by high cost, inconvenience in use and relatively serious side effects. In traditional Chinese medicine, such an effective drug is also missing. Due to the complexity of the manifestations of the metabolic syndrome, no drug specifically designed for the treatment of metabolic syndrome has been presented in the world to date, therefore, clinical studies in order to develop new drugs for the treatment of metabolic syndrome are of great importance. The present invention fills the gap in this area in the world.

SHORT DESCRIPTION

One of the objectives of the present invention is to provide a Traditional Chinese Medicine medicinal material composition, wherein the Traditional Chinese Medicine medicinal material composition comprises Coptidis Rhizoma, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Trichosanthis Fructus, Cinnamomi Cortex, and Red Yeast Rice.

In the composition, Aurantii Fructus Immaturus is preferably bran roasted Aurantii Fructus Immaturus, Pinelliae Rhizoma is preferably Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine, and Trichosanthis Fructus is preferably whole Trichosanthis Fructus. Preferably the composition contains 5-80 wt. % Coptidis Rhizoma, 5-70 wt. % Aurantii Fructus Immaturus, 0-60 wt. % Pinelliae Rhizoma, 0-40 wt. % Trichosanthis Fructus, 2-12 wt. % Cinnamomi Cortex and 10-60 wt. % red yeast rice. More preferably, the composition contains 10-30 wt. % Coptidis Rhizoma, 10-30 wt. % Aurantii Fructus Immaturus, 10-30 wt. % Pinelliae Rhizoma, 5-30 wt. % Trichosanthis Fructus, 2-10 wt. % Cinnamomi Cortex and 12-36 wt. % red yeast rice. Most preferably, the preferred proportions of the components in the composition are as follows: 19.6 wt. % Coptidis Rhizoma, 19.6 wt. % Aurantii Fructus Immaturus, 19.6 wt. % Pinelliae Rhizoma, 26.1 wt. % Trichosanthis Fructus, 2.1 wt. % Cinnamomi Cortex and 13.0 wt. % red yeast rice.

The second object of the present invention is to provide a composition based on medicinal materials of traditional Chinese medicine, where the composition contains Coptidis Rhizoma, Aurantii Fructus Immaturus, red yeast rice and optionally one or more components selected from the group consisting of Pinelliae Rhizoma, Trichosanthis Fructus and Cinnamomi Cortex. Said one or more components include 1, 2, 3 components.

The composition contains 5-80 wt. % Coptidis Rhizoma; 5-70 wt. % Aurantii Fructus Immaturus; 0-60 wt. % Pinelliae Rhizoma; 0-40 wt. % Trichosanthis Fructus; 0-12 wt. % Cinnamomi Cortex and 10-60 wt. % red yeast rice.

The composition contains 5-80 wt. % Coptidis Rhizoma; 5-70 wt. % Aurantii Fructus Immaturus; 0-60 wt. % Pinelliae Rhizoma; 5-40 wt. % Trichosanthis Fructus; 2-12 wt. % Cinnamomi Cortex and 10-60 wt. % red yeast rice.

The composition contains 5-80 wt. % Coptidis Rhizoma; 5-70 wt. % Aurantii Fructus Immaturus; 10-60 wt. % Pinelliae Rhizoma; 0-40 wt. % Trichosanthis Fructus; 2-12 wt. % Cinnamomi Cortex and 10-60 wt. % red yeast rice.

The composition contains 5-80 wt. % Coptidis Rhizoma; 5-70 wt. % Aurantii Fructus Immaturus; 10-60 wt. % Pinelliae Rhizoma; 5-40 wt. % Trichosanthis Fructus; 0-12 wt. % Cinnamomi Cortex and 10-60 wt. % red yeast rice.

The composition contains 5-80 wt. % Coptidis Rhizoma; 5-70 wt. % Aurantii Fructus Immaturus; 0-60 wt. % Pinelliae Rhizoma; 0-40 wt. % Trichosanthis Fructus; 2-12 wt. % Cinnamomi Cortex and 10-60 wt. % red yeast rice.

The composition contains 5-80 wt. % Coptidis Rhizoma; 5-70 wt. % Aurantii Fructus Immaturus; 0-60 wt. % Pinelliae Rhizoma; 5-40 wt. % Trichosanthis Fructus; 0-12 wt. % Cinnamomi Cortex and 10-60 wt. % red yeast rice.

The composition contains 5-80 wt. % Coptidis Rhizoma; 5-70 wt. % Aurantii Fructus Immaturus; 10-60 wt. % Pinelliae Rhizoma; 0-40 wt. % Trichosanthis Fructus; 0-12 wt. % Cinnamomi Cortex and 10-60 wt. % red yeast rice.

Aurantii Fructus Immaturus is preferably bran roasted Aurantii Fructus Immaturus, Pinelliae Rhizoma is preferably Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine, and Trichosanthis Fructus is preferably whole Trichosanthis Fructus.

Another object of the present invention is to provide a composition based on extracts of medicinal materials of traditional Chinese medicine obtained by mixing extracts of six medicinal materials of traditional Chinese medicine, consisting of Coptidis Rhizoma, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Trichosanthis Fructus, Cinnamomi Cortex and red yeast rice. The relative content of the extracts is as follows: 2-30% is the total alkaloid extract of Coptidis Rhizoma (abbreviated name of Coptidis Rhizoma extract), and 70-98% is the mixed extract of five medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice (from the total amount in the composition, 10-100% of red yeast rice is extracted, 0-90% is ground into a fine powder for use as a baking powder or filler, hereinafter by analogy), Trichosanthis Fructus and Cinnamomi Cortex. Preferably, the extract of Coptidis Rhizoma is 3-25%, and the mixed extract of five medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice, Trichosanthis Fructus and Cinnamomi Cortex, is 75-97%. Most preferably, the extract of Coptidis Rhizoma is 5-15%, and the mixed extract of five medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice, Trichosanthis Fructus and Cinnamomi Cortex, is 85-95%.

Red yeast rice powder (0-90% of the total composition) and Coptidis Rhizoma extract are mixed until homogeneous with a mixed extract of five medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice, Trichosanthis Fructus and Cinnamomi Cortex, or the corresponding extracts of five medicinal materials of traditional Chinese medicine to obtain a composition based on extracts of medicinal materials of traditional Chinese medicine.

Composition based on extracts of medicinal materials of traditional Chinese medicine contains the following active ingredients (mg/g): 6-122 mg/g of total alkaloids (the total amount of four types of alkaloids: berberine hydrochloride, jatrorrizine hydrochloride, coptisine hydrochloride and palmatin hydrochloride, further by analogy) , including 4-89 mg/g berberine; 30-315 mg/g of total flavonoids (the total amount of three flavonoid glycosides, including neohesperidin, hesperidin and naringin, further by analogy), including 20-186 mg/g of neohesperidin, and 0.5-9 mg/g of lovastatin.

Composition based on extracts of medicinal materials of traditional Chinese medicine contains the following active ingredients (mg/g): 12-122 mg/g total alkaloids, including 9-89 mg/g berberine; 130-315 mg/g total flavonoids, including 80-186 mg/g neohesperidin, and 1-9 mg/g lovastatin.

The composition based on extracts of medicinal materials of traditional Chinese medicine preferably contains the following active ingredients (mg/g): 20-85 mg/g total alkaloids, including 15-62 mg/g berberine; 140-250 mg/g total flavonoids, including 90-170 mg/g neohesperidin, and 1-5 mg/g lovastatin.

The composition based on extracts of medicinal materials of traditional Chinese medicine more preferably contains the following active ingredients (mg/g): 30-75 mg/g total alkaloids, including 20-50 mg/g berberine; 110-250 mg/g total flavonoids, including 60-120 mg/g neohesperidin, and 1-5 mg/g lovastatin.

The composition based on extracts of medicinal materials of traditional Chinese medicine preferably contains the following active ingredients (mg/g): 20-85 mg/g total alkaloids, including 15-62 mg/g berberine; 100-250 mg/g total flavonoids, including 30-170 mg/g neohesperidin, and 1-5 mg/g lovastatin.

The composition based on extracts of medicinal materials of traditional Chinese medicine more preferably contains the following active ingredients (mg/g): 30-75 mg/g total alkaloids, including 20-50 mg/g berberine; 110-250 mg/g total flavonoids, including 40-120 mg/g neohesperidin, and 1-5 mg/g lovastatin.

Yet another object of the present invention is to provide a method for producing a traditional Chinese medicine extract. Coptidis Rhizoma is extracted using an aqueous acid solution and filtered, concentrated, alkalinely precipitated, salted out, filtered, dried and ground to obtain a total alkaloid extract. Four drug materials, including red yeast rice (10-100% of the amount required by the prescription), Aurantii Fructus Immaturus, Pinelliae Rhizoma and Cinnamomi Cortex, are extracted using 50-90% ethanol and filtered. Trichosanthis Fructus is extracted using 75-95% ethanol, the Trichosanthis Fructus extract solution is combined with the extract solution of four medicinal materials of traditional Chinese medicine, the ethanol is removed, concentrated under reduced pressure, dried and pulverized to obtain a mixed extract of five medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, Red Yeast Rice, Cinnamomi Cortex and Trichosanthis Fructus; or five medicinal materials of traditional Chinese medicine, including red yeast rice, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Cinnamomi Cortex and Trichosanthis Fructus, are mixed and extracted using 50-90% ethanol, filtered, ethanol removed under reduced pressure, concentrated, dried and crushed to obtain a mixed extract of five medicinal materials of traditional Chinese medicine, consisting of red yeast rice, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Cinnamomi Cortex and Trichosanthis Fructus; or red yeast rice, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Cinnamomi Cortex and Trichosanthis Fructus are separately extracted using 50-90% ethanol, filtered to obtain separately solutions of ethanol extracts of individual medicinal materials of traditional Chinese medicine; the separately obtained solutions of ethanol extracts of the five medicinal materials of traditional Chinese medicine are separately filtered, the ethanol is removed under reduced pressure, and concentrated, dried and pulverized to obtain the corresponding extracts of the five medicinal materials of traditional Chinese medicine, consisting of red yeast rice, Aurantii Fructus Immaturus, Pinelliae Rhizoma , Cinnamomi Cortex and Trichosanthis Fructus. The respective extracts of the five medicinal materials of traditional Chinese medicine are mixed to obtain a mixed extract of the five medicinal materials of traditional Chinese medicine. The extract of Coptidis Rhizoma and the mixed extract of the five medicinal materials of traditional Chinese medicine are mixed to obtain a composition based on extracts of medicinal materials of traditional Chinese medicine.

In the composition, Coptidis Rhizoma is extracted with 0.1-1% aqueous sulfuric acid, filtered, the filtrate is concentrated, calcium hydroxide is added for alkaline precipitation, the precipitate is removed, the pH of the filtrate is adjusted with hydrochloric acid to an acidic pH, then sodium chloride is added to precipitate the precipitate and the precipitate are separated, washed, filtered, dried and ground to obtain an extract of Coptidis Rhizoma.

In the composition, four drug materials, including red yeast rice (10-100% of the amount required by the prescription), Aurantii Fructus Immaturus, Pinelliae Rhizoma and Cinnamomi Cortex, are extracted using 50-90% ethanol and filtered; and Trichosanthis Fructus are extracted using 75-95% ethanol, the Trichosanthis Fructus extract solution is combined with the extract solution of the four medicinal materials of traditional Chinese medicine, the ethanol is removed, concentrated under reduced pressure, dried and pulverized to obtain a mixed extract of the five medicinal materials of traditional Chinese medicine, consisting from Aurantii Fructus Immaturus, Pinelliae Rhizoma, Red Yeast Rice, Cinnamomi Cortex and Trichosanthis Fructus.

In the composition, five medicinal materials of traditional Chinese medicine, including red yeast rice, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Trichosanthis Fructus and Cinnamomi Cortex, are mixed and extracted using 50-90% ethanol, filtered, ethanol removed under reduced pressure, concentrated, dried and crushed to obtain a mixed extract of five medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice, Cinnamomi Cortex and Trichosanthis Fructus. Alternatively, red yeast rice, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Cinnamomi Cortex, and Trichosanthis Fructus are separately extracted using 50-90% ethanol, filtered to obtain separately solutions of ethanol extracts of selected medicinal materials of traditional Chinese medicine; the separately obtained solutions of ethanol extracts of the five medicinal materials of traditional Chinese medicine are separately filtered, the ethanol is removed under reduced pressure, and concentrated, dried, pulverized and mixed to obtain individual extracts of the five medicinal materials of traditional Chinese medicine, consisting of red yeast rice, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Cinnamomi Cortex and Trichosanthis Fructus. In ethanol extraction of five traditional Chinese medicine medicinal materials including Aurantii Fructus Immaturus, Pinelliae Rhizoma, Cinnamomi Cortex, red yeast rice and Trichosanthis Fructus to obtain a mixed extract or single extracts of traditional Chinese medicine medicinal materials, the ethanol is preferably 60-85% ethanol.

The mixed extract of the five medicinal materials of traditional Chinese medicine, or the corresponding extracts of the five medicinal materials of traditional Chinese medicine, or the extract of Trichosanthis Fructus and the extracts of the four medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice, Cinnamomi Cortex, are mixed with extract of Coptidis Rhizoma and with 0-90% fine powder of red yeast rice used as a disintegrant or tablet filler to obtain a composition based on extracts of medicinal materials of traditional Chinese medicine.

Another object of the present invention is to provide a composition based on medicinal materials of traditional Chinese medicine, providing an administration dose.

The daily dose of administration of the composition based on medicinal materials of traditional Chinese medicine according to the present invention is as follows: 1.5-9 g Coptidis Rhizoma, 1.5-9 g Aurantii Fructus Immaturus, 0-12 g Pinelliae Rhizoma, 0-12 g Trichosanthis Fructus , 0-5 g Cinnamomi Cortex and 1.5-30 g red yeast rice. Preferably the dose is 3-6 g Coptidis Rhizoma, 3-6 g Aurantii Fructus Immaturus, 3-6 g Pinelliae Rhizoma, 0-6 g Trichosanthis Fructus, 0.5-2.5 g Cinnamomi Cortex and 1.5-6 g red yeast rice. The most preferred dose is 4.5 g Coptidis Rhizoma, 4.5 g Aurantii Fructus Immaturus, 4.5 g Pinelliae Rhizoma, 6 g Trichosanthis Fructus, 0.5 g Cinnamomi Cortex and 3 g red yeast rice.

Another object of the present invention is to provide a composition based on extracts of medicinal materials of traditional Chinese medicine, providing an administration dose.

The dose of administration of the composition based on extracts of medicinal materials of traditional Chinese medicine according to the present invention is 1-15 g per day; the preferred dose of administration is 2-10 g per day, and more preferably the dose of administration is 3-6 g per day.

Composition based on extracts of medicinal materials of traditional Chinese medicine contains the following active ingredients (mg/g): 6-122 mg/g total alkaloids, including 4-89 mg/g berberine; 30-315 mg/g total flavonoids, including 20-186 mg/g neohesperidin, and 0.5-9 mg/g lovastatin.

In some examples, the composition based on extracts of medicinal materials of traditional Chinese medicine contains the following active ingredients (mg/g): 12-122 mg/g total alkaloids, including 9-89 mg/g berberine; 130-315 mg/g total flavonoids, including 80-186 mg/g neohesperidin, and 1-9 mg/g lovastatin.

In some examples, the composition based on extracts of medicinal materials of traditional Chinese medicine preferably contains the following active ingredients (mg/g): 20-85 mg/g of total alkaloids, including 15-62 mg/g berberine; 100-250 mg/g total flavonoids, including 30-170 mg/g neohesperidin, and 1-5 mg/g lovastatin. In some examples, the composition based on extracts of medicinal materials of traditional Chinese medicine more preferably contains the following active ingredients (mg/g): 30-75 mg/g total alkaloids, including 20-50 mg/g berberine; 110-250 mg/g total flavonoids, including 40-120 mg/g neohesperidin, and 1-5 mg/g lovastatin.

In some examples, the composition based on extracts of medicinal materials of traditional Chinese medicine preferably contains the following active ingredients (mg/g): 20-85 mg/g of total alkaloids, including 15-62 mg/g berberine; 140-250 mg/g total flavonoids, including 90-170 mg/g neohesperidin, and 1-5 mg/g lovastatin. In some examples, the composition based on extracts of medicinal materials of traditional Chinese medicine more preferably contains the following active ingredients (mg/g): 30-75 mg/g total alkaloids, including 20-50 mg/g berberine; 110-250 mg/g total flavonoids, including 60-120 mg/g neohesperidin, and 1-5 mg/g lovastatin.

DESCRIPTION OF THE INVENTION

Overview of current research

Coptidis Rhizoma. The main components of Coptidis Rhizoma are berberine hydrochloride, coptisine hydrochloride and palmatine hydrochloride, and the like. Coptidis Rhizoma helps lower blood glucose levels and lower serum lipid levels, has antioxidant, antibacterial and antiviral effects.

Aurantii Fructus Immaturus. The main components of Aurantii Fructus Immaturus are flavonoids and alkaloids. The main active ingredients of Aurantii Fructus Immaturus are naringin, hesperidin, neohesperidin and synephrine. Aurantii Fructus Immaturus improves the function of the gastrointestinal tract, has an antithrombotic effect and enhances myocardial contraction.

Trichosanthis Fructus. The main active ingredients of Trichosanthis Fructus are flavonoids and saponins. Trichosanthis Fructus has an expectorant and antibacterial effect, prevents the development of coronary heart disease, and has a rejuvenating effect.

Pinelliae Rhizoma. The main active ingredients of Pinelliae Rhizoma are 6-gingerol and alkaloids and the like. Pinelliae Rhizoma helps lower blood pressure and has an antitussive effect.

Cinnamomi Cortex. The main components of Cinnamomi Cortex are cinnamic acid and cinnamaldehyde. Cinnamomi Cortex has a calming effect, lowers blood pressure.

Red Yeast Rice: The main active ingredient in red yeast rice is lovastatin. Red yeast rice lowers blood lipids, has a non-specific immune-enhancing effect, can increase the phagocytic activity of phagocytes, enhance antibody production, and improve T cell function.

Explanation of the traditional recipe

Metabolic syndrome is caused by eating fatty and high-calorie foods, phlegm that turns into heat, which fight each other and condense in the middle part of the body. The Coptidis Rhizoma contained in the composition has a bitter taste and a cooling effect, has a tropism for the meridians of the heart, stomach and large intestine. It has the function of eliminating heat and removing fire, getting rid of stagnation and eliminating bloating, eliminating stomach fever syndrome and quenching thirst. In the book Yao Lei Fa Xiang, Coptidis Rhizoma is described as “a remedy for removing the fire of the heart, removing moisture and heat in the spleen and stomach, curing irritability and nausea, getting rid of stagnation of heat in the middle part of the body, and vomiting. It is an important remedy for the treatment of induration in the epigastric region." Thickness in the epigastric region refers to the symptom of "gastric distention, feeling of pain when pressed", described in the ancient recipe "Xiaoxianxiong Decoction" by doctor Zhong Jing, while their manifestations are similar to the symptom of abdominal distention in metabolic syndrome. Thus, this component is used as the "main" drug.

Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine has a warm character and a warming effect and affinity for the meridians of the spleen, stomach and lungs. It eliminates phlegm, disperses phlegm in the abdominal cavity and eliminates congestion in the stomach, regulates Qi and eliminates vomiting. Pinelliae Rhizoma is described in the book "Ming Yi Bie Lu" as "a remedy for expectoration and heat in the chest and hypochondrium, cough, induration and hardness in the epigastric part with a feeling of pain, and also vomiting." When Pinelliae Rhizoma is used with Coptidis Rhizoma, one is pungent and the other bitter, the pungent remedy has a dispersing effect and the bitter remedy has a laxative effect, both enter the middle part of the body, remove heat and eliminate phlegm, disperse indurations and eliminate turgor. Trichosanthis Fructus has a bitter taste and a cooling effect, affinity for the meridians of the lungs and stomach. It has the functions of removing heat and dispelling phlegm, eliminating rheumatic pain and eliminating turgor, improving Qi circulation and moistening the intestines. Metabolic syndrome patients have a hard and bloated abdomen and Trichosanthis Fructus can remove the bloating. Pinelliae Rhizoma is well compatible with Trichosanthis Fructus, both remedies can relieve dryness, eliminate heat and remove phlegm. Trichosanthis Fructus also has good compatibility with Coptidis Rhizoma, both remedies work well on the middle part of the body, bitterness eliminates dryness, promotes a curative effect and removes stagnant phlegm in the middle part of the body. The combination of these three drugs is known from the Xiaoxianxiong Decoction prescription. Aurantii Fructus Immaturus fried in bran has a bitter taste and a cooling effect, affinity for the meridians of the lungs and stomach. It improves the circulation of qi in the abdomen and eliminates phlegm in the body. Aurantii Fructus Immaturus is described in the book Ming Yi Bie Lu as "a remedy for eliminating phlegm in the chest and hypochondrium, removing fluid stagnation, dispelling Qi stagnation and swelling, reducing swelling and pain in the epigastric region, lowering ascending Qi." The book "Yao Pin Hua Yi" states that Aurantii Fructus Immaturus promotes digestion and eliminates the feeling of fullness in the stomach, eliminates constipation, promotes hematopoiesis, eliminates bloating and mucus accumulation, removes fluid retention, eliminates undigested food and relieves constipation. Patients with metabolic syndrome have a hard and bloated abdomen, Aurantii Fructus Immaturus seems to be appropriate. Thus, all of Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine, Trichosanthis Fructus, Aurantii Fructus Immaturus are all remedies "increasing the action of the master remedy." The bitterness of Coptidis Rhizoma has a cleansing effect, the pungency of Pinelliae Rhizoma has a dispersing effect, the bitterness of Trichosanthis Fructus moisturizes the colon, and the bitterness of Aurantii Fructus Immaturus promotes Qi circulation. The Chinese healer Zhang Yuansu said: "A combination of Aurantii Fructus Immaturus and Coptidis Rhizoma should be used to treat bloating in the epigastric region and indigestion."

Red yeast rice is non-toxic and has a sweet and warm character. It has a tropism for the meridians of the liver, spleen, stomach and intestines. It promotes digestion and normalizes the function of the stomach and is good for removing fat, activating blood circulation and dispels blood stasis. The book "Ben Cao Yan Yi Bu Yi" says that red yeast rice "stimulates blood circulation and promotes digestion, strengthens the spleen and warms the stomach." The book Ben Cao Qiu Yuan states that red yeast rice is “made from short-grain rice with the addition of wine sourdough to give it a red color. It has the effect of stimulating blood circulation by improving Qi circulation, and also improves stomach function and promotes digestion. It can be used to treat anyone who suffers from excessive emotionality due to clotting Qi.” Thus, red yeast rice is used as an "auxiliary" drug.

The spicy taste and warming effect of Cinnamomi Cortex helps to eliminate Qi stagnation in the middle part of the body. Cinnamomi Cortex is described in Yu Qiu Yao Jie as follows: “Cinnamomi Cortex warms the stomach, increases Qi circulation, tones and warms Qi in the blood. It has a fragrant aroma and sweet taste, which contributes to its effect on the spleen meridian; the sharpness, which has a dispersing effect, and the sweet taste help to eliminate depression and stagnation of qi in the liver and spleen. When used with Cinnamomi Cortex along with Coptidis Rhizoma, the pungent medicine has a dispersing effect and the bitter medicine has a laxative effect, they remove heat and protect the spleen, their combination prevents the excessive cooling effect of Coptidis Rhizoma, which suppresses the function of the spleen and stomach, and also helps to dispel congestion and promotes fluid production in the body. Thus, Cinnamomi Cortex is used as a "drug guide".

All these products are compatible, when used together as a whole, they have the effect of removing heat, dispersing phlegm and stimulating the circulation of Qi to help eliminate abdominal distention. They are used to treat symptoms caused by sputum stasis and fever, such as obesity, enlarged abdomen, chest tightness, copious sputum, lack of appetite and a feeling of fullness in the stomach and abdomen, dry mouth, thirst, polydipsia and persistent feeling of slight hunger, irritability. , hot flashes to the face and oily skin of the face, night snoring, loose stools or constipation, bright yellow urine, red tongue with a yellow coating and unstable pulse. In Western medicine, they can also be used to treat patients with hyperlipidemia, diabetes mellitus, arterial hypertension, obesity, and metabolic syndrome (abdominal obesity with abnormalities in blood sugar, glycated hemoglobin, blood pressure, blood lipids, and uric acid). Efficacy testing was performed using spontaneous type 2 diabetic KKAy mice with symptoms of insulin resistance and obesity and C57BL/6J mice fed a high fat diet as experimental animal models. Accordingly, 500, 1000 and 1500 mg of the traditional Chinese medicine medicinal material composition of the present invention (corresponding to the intake of 3000, 6000 and 9000 mg of the traditional Chinese medicine medicinal material composition of the present invention by an adult) were administered in order to investigate the effect of the composition based on medicinal materials of traditional Chinese medicine of the present invention on indicators related to metabolic syndrome, such as insulin resistance, carbohydrate and lipid metabolism, blood pressure. The results showed that the traditional Chinese medicine medicinal material composition of the present invention has the following effects: (1) lowering blood glucose; (2) decrease in the level of insulin in the blood; (3) improvement in glucose tolerance as measured by an oral test; (4) decrease in the level of glycated hemoglobin (HbA1c); (5) improving insulin sensitivity index and improving insulin resistance; and (6) lowering blood triglycerides, trending lower total cholesterol and lowering blood pressure. Thus, the inventors of the present invention determined that the preferred daily dose of administration of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention is 2-10 g.

Drug stability testing showed that the traditional Chinese medicine medicinal material composition tablets of the present invention readily absorb moisture and are more sensitive to high humidity than to high temperature and light; exposure to high temperature leads to a significant decrease in the content of cinnamaldehyde; and exposure to light leads to a significant decrease in the content of lovastatin. In terms of final packaging for tablets, an inner packaging in the form of aluminum-plastic blisters is used, which are packed in an aluminum foil package. Conducted an accelerated test at a temperature of 40°C±2°C and a relative humidity of 75±5% for 6 months. The weight change rate due to moisture absorption of the tablets with the composition of traditional Chinese medicine medicinal materials of the present invention was in the range of 0.5%, which shows a good waterproofing and moisture-proofing effect of this package. The amounts of berberine hydrochloride, total alkaloids from Coptidis Rhizoma, total flavonoids and naringin from Aurantii Fructus Immaturus in the traditional Chinese medicine medicinal material composition tablet of the present invention change very little; the amount of lovastatin slightly changes, while the degree of change of the content is within 12.23%, and the change is mainly due to the interconversion of isomers and does not affect the efficacy.

Through stability studies of tablets with the composition based on medicinal materials of traditional Chinese medicine of the present invention, it was confirmed that an aluminum-plastic blister pack should be used as the inner package; storage conditions should provide for sealed storage in a dry place. Under the above storage conditions, the final product is stable, moisture protection is effective, and the shelf life is provisionally set to two years.

One of the objectives of the present invention is to provide a composition based on medicinal materials of traditional Chinese medicine, which has an accurate list of active ingredients, a mechanism of action, well-defined pharmacological activity and clinical efficacy, controlled quality and safety of administration.

The technical solution used in the present invention is as follows: a composition based on medicinal materials of traditional Chinese medicine containing Coptidis Rhizoma, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Trichosanthis Fructus, Cinnamomi Cortex and red yeast rice.

In the composition, Aurantii Fructus Immaturus is preferably bran roasted Aurantii Fructus Immaturus, Pinelliae Rhizoma is preferably Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine, and Trichosanthis Fructus is preferably whole Trichosanthis Fructus.

Preferably the composition contains 5-80 wt. % Coptidis Rhizoma, 5-70 wt. % Aurantii Fructus Immaturus, 0-60 wt. % Pinelliae Rhizoma, 0-40 wt. % Trichosanthis Fructus, 2-12 wt. % Cinnamomi Cortex and 10-60 wt. % red yeast rice. More preferably, the composition contains 10-30 wt. % Coptidis Rhizoma, 10-30 wt. % Aurantii Fructus Immaturus, 10-30 wt. % Pinelliae Rhizoma, 5-30 wt. % Trichosanthis Fructus, 2-10 wt. % Cinnamomi Cortex and 12-36 wt. % red yeast rice. Most preferably, the preferred proportions of the components in the composition are as follows: 19.6 wt. % Coptidis Rhizoma, 19.6 wt. % Aurantii Fructus Immaturus, 19.6 wt. % Pinelliae Rhizoma, 26.1 wt. % Trichosanthis Fructus, 2.1 wt. % Cinnamomi Cortex and 13.0 wt. % red yeast rice.

Another technical solution provided by the present invention is as follows: a composition containing Coptidis Rhizoma, Aurantii Fructus Immaturus, red yeast rice and one or more components selected from the group consisting of Pinelliae Rhizoma, Trichosanthis Fructus and Cinnamomi Cortex.

The composition contains 5-80 wt. % Coptidis Rhizoma; 5-70 wt. % Aurantii Fructus Immaturus; 0-60 wt. % Pinelliae Rhizoma; 0-40 wt. % Trichosanthis Fructus; 0-12 wt. % Cinnamomi Cortex and 10-60 wt. % red yeast rice.

The dose of administration of a composition based on medicinal materials of traditional Chinese medicine is 10-50 g per day; preferably the dose of administration is 14-38 g per day; most preferably, the administration dose is 18-25 g per day.

In another aspect, the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention is obtained by mixing extracts of six medicinal materials of traditional Chinese medicine: Coptidis Rhizoma, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Trichosanthis Fructus, Cinnamomi Cortex and red yeast rice. The relative content of the extracts is as follows: Coptidis Rhizoma extract is 2-30%, and a mixed extract of the five medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice (including 10-100% of which is extracted, 0- 90% is ground into a fine powder for use as a baking powder or filler, hereinafter by analogy), Trichosanthis Fructus and Cinnamomi Cortex, is 70-98%. Preferably, the extract of Coptidis Rhizoma is 3-25%, and the mixed extract of five medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice, Trichosanthis Fructus and Cinnamomi Cortex, is 75-97%. More preferably, the extract of Coptidis Rhizoma is 5-15%, and the mixed extract of five medicinal materials of traditional Chinese medicine, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice, Trichosanthis Fructus and Cinnamomi Cortex, is 85-95%.

Composition based on extracts of medicinal materials of traditional Chinese medicine contains the following active ingredients (mg/g): 6-122 mg/g total alkaloids, including 4-89 mg/g berberine; 30-315 mg/g total flavonoids, including 20-186 mg/g neohesperidin, and 0.5-9 mg/g lovastatin. Preferably, the composition based on extracts of medicinal materials of traditional Chinese medicine contains the following active ingredients (mg/g): 20-85 mg/g of total alkaloids, including 15-62 mg/g of berberine; 100-250 mg/g total flavonoids, including 30-170 mg/g neohesperidin, and 1-5 mg/g lovastatin. More preferably, the composition based on extracts of medicinal materials of traditional Chinese medicine contains the following active ingredients (mg/g): 30-75 mg/g of total alkaloids, including 20-50 mg/g of berberine; 110-250 mg/g total flavonoids, including 40-120 mg/g neohesperidin, and 1-5 mg/g lovastatin.

Composition based on extracts of medicinal materials of traditional Chinese medicine contains the following active ingredients (mg/g): 12-122 mg/g total alkaloids, including 9-89 mg/g berberine; 130-315 mg/g total flavonoids, including 80-186 mg/g neohesperidin, and 1-9 mg/g lovastatin. Preferably, the composition based on extracts of medicinal materials of traditional Chinese medicine contains the following active ingredients (mg/g): 20-85 mg/g of total alkaloids, including 15-62 mg/g of berberine; 140-250 mg/g total flavonoids, including 90-170 mg/g neohesperidin, and 1-5 mg/g lovastatin. More preferably, the composition based on extracts of medicinal materials of traditional Chinese medicine contains the following active ingredients (mg/g): 30-75 mg/g of total alkaloids, including 20-50 mg/g of berberine; 110-250 mg/g total flavonoids, including 60-120 mg/g neohesperidin, and 1-5 mg/g lovastatin.

The dose of administration of a composition based on extracts of medicinal materials of traditional Chinese medicine is 1-15 g per day; preferably the dose of administration is 2-10 g per day; most preferably, the administration dose is 3-6 g per day.

The traditional Chinese medicine material composition and the traditional Chinese medicine material extract composition can be prepared as pharmaceutical preparations such as injectables, tablets, capsules, granules, and oral solutions.

The traditional Chinese medicine medicinal material composition and the traditional Chinese medicine medicinal material extract composition have the effect of removing heat and dispersing phlegm, stimulating Qi circulation, and helping to eliminate bloating. The compositions are used to treat symptoms caused by sputum stasis and fever, such as obesity, enlarged abdomen, chest tightness, copious sputum, lack of appetite and feeling of fullness in the stomach and abdomen, dry mouth, thirst, polydipsia and persistent feeling of slight hunger, irritability , hot flashes to the face and oily skin of the face, night snoring, loose stools or constipation, bright yellow urine, red tongue with a yellow coating and unstable pulse. In Western medicine, the composition can also treat symptoms including hyperlipidemia, diabetes mellitus, arterial hypertension, obesity, and metabolic syndrome (abdominal obesity with abnormalities in blood sugar, glycated hemoglobin, blood pressure, blood lipids, and uric acid).

The traditional Chinese medicine medicinal material composition and/or the traditional Chinese medicine medicinal material extract composition can be used for the preparation of medicaments for the treatment of one or more of the following symptoms: metabolic syndrome, obesity, hyperglycemia, high blood pressure, or hyperlipidemia.

In some examples, drugs are used to treat one or more of the following disorders: blood glucose and lipid metabolism disorders, obesity, hyperglycemia, hypertension, and hyperlipidemia in the early stages of the metabolic syndrome. In some examples, drugs are used to provide one or more of the following effects: lowering blood glucose levels, lowering blood insulin levels, improving oral glucose tolerance, lowering glycated hemoglobin (HbA1c), improving insulin sensitivity index, improve insulin resistance, lower blood triglycerides, lower total cholesterol and lower blood pressure.

Several kinds of effects include 2, 3, 4, 5, 6, 7, 8, 9 effects.

The composition based on extracts of traditional Chinese medicine medicinal materials of the present invention has the following beneficial technical effects.

(1) Significant improvement in the treatment of metabolic syndrome.

The results of the comparative pharmacodynamic test of the extracts obtained according to examples 1-7 indicate that the extracts of the present invention have statistically different effects in reducing blood sugar, triglycerides and total cholesterol in the blood serum. Systemic pharmacodynamic studies confirmed that the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention has the following effects: body weight reduction, blood sugar reduction, blood insulin reduction, improvement of glucose tolerance by oral test, reduction of glycated hemoglobin (HbA1c), increase in insulin sensitivity index, improvement in insulin resistance, decrease in blood triglycerides, decrease in total cholesterol, lower blood pressure.

(2) Stable properties and controlled quality of the composition based on medicinal materials of traditional Chinese medicine of the present invention.

Through stability studies of tablets with the composition based on medicinal materials of traditional Chinese medicine of the present invention, it was confirmed that an aluminum-plastic blister pack should be used as the inner package; storage conditions should provide for sealed storage in a dry place. Under the above storage conditions, the final product is stable, moisture protection is effective, and the shelf life is provisionally set to two years.

(3) Safe use of the traditional Chinese medicine composition of the present invention.

When tested for acute toxicity in mice that were orally administered with the composition based on medicinal materials of traditional Chinese medicine of the present invention, it was shown that the value of LD ₅₀ is more than 20000 mg/kg, which is 200 times the expected clinical dose. Acute toxicity testing in beagle dogs has shown a lethal dose of over 7500 mg/kg, 75 times the expected clinical dose.

When tested for long-term toxicity in rats, the doses of the traditional Chinese medicine medicinal material composition of the present invention were 8 times, 28 times, and 98 times the intended clinical dose, and the results showed that the expected clinical dose of the traditional Chinese medicine medicinal material composition according to the present invention is safe.

Suitability of the composition based on medicinal materials of traditional Chinese medicine according to the present invention for industrial production. The results of pilot production of 9 batches showed the feasibility of the production technology.

DETAILED DESCRIPTION OF EMBODIMENTS

The following embodiments are used to illustrate the invention, but not to limit the scope of the present invention. If a specific method or conditions are not described in the embodiments, they are carried out in accordance with the method or conditions described in the relevant literature in this field, or in accordance with the instructions for use of the product. All reagents and instruments for which no manufacturer is listed are regular products that can be purchased through official channels.

Example 1

Composition 1: 1.5 g (7.9 wt%) Coptidis Rhizoma, 6 g (31.6 wt%) Aurantii Fructus Immaturus, 3 g (15.8 wt%) Pinelliae Rhizoma, 0 g (0 wt. %) Trichosanthis Fructus, 2.5 g (13.1 wt. %) Cinnamomi Cortex and 6 g (31.6 wt. %) red yeast rice, with a total daily dose of 19 g. The amount of extracts obtained is 5 .58 g, with a yield of 29.4%.

Production method: Coptidis Rhizoma was crushed to a coarse powder or medicinal raw materials were cut into pieces in the form of pieces of a suitable size and placed in an extractor; 0.5% sulfuric acid was added in a 10-fold amount, three cycles of extraction by evaporation were performed, the duration of each cycle was 2 hours; after which filtration was carried out, the solutions of the extracts were combined, the pressure was reduced for concentration; the concentrated solution was placed in a settling vessel and stirred, milk of lime was slowly added with pH adjusted to 9, settled, filtered to remove calcium sulfate precipitate; the pH value of the filtrate was adjusted with concentrated hydrochloric acid to a value of 1-2, sodium chloride was added to 10% by volume, salted out, the solution was settled for 24 hours, filtered with suction; the precipitate was washed with water until pH 5 was reached and dried under reduced pressure at 60°C to obtain an extract of the total alkaloids.

The following medicinal materials: bran fried Aurantii Fructus Immaturus, Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine, red yeast rice and Cinnamomi Cortex, crushed to a coarse powder or cut medicinal raw materials in the form of pieces into pieces of suitable size, medicinal materials were obtained according to the required amount according to the prescription, mixed and placed in a reverse flow extractor using ethanol, and performed two extraction cycles with 70% ethanol 10 times and with 70% ethanol 8 times, respectively, the duration of each cycle is 1 hour; filtered, removed the ethanol under reduced pressure, concentrated and dried to obtain a mixed extract of four medicinal materials of traditional Chinese medicine.

The blended extract of four traditional Chinese medicine medicinal materials, consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, Cinnamomi Cortex, and red yeast rice, is mixed with Coptidis Rhizoma extract until homogeneous to obtain the final composition based on extracts of traditional Chinese medicine medicinal materials of the present invention.

Content determination result (mg/g): total alkaloid content is 12.53 mg/g (including 9.27 mg/g berberine), total flavonoid content is 292.48 mg/g (including 171.11 mg /g of neohesperidin), and the content of lovastatin is 4.31 mg/g.

Example 2

Composition 2: 4.5 g (45 wt%) Coptidis Rhizoma, 3 g (30 wt%) Aurantii Fructus Immaturus, 0 g (0 wt%) Pinelliae Rhizoma, 0.5 g (5 wt%) Trichosanthis Fructus, 0.5 g (5 wt%) Cinnamomi Cortex and 1.5 g (15 wt%) red yeast rice; while the total daily dose of administration is 10 g. The amount of extracts obtained is 2.74 g, while the yield is 27.4%.

Method of preparation: an extract of the total alkaloids from Coptidis Rhizoma was obtained using the method of example 1.

The following medicinal materials: Bran-fried Aurantii Fructus Immaturus, Red Yeast Rice, Trichosanthis Fructus and Cinnamomi Cortex were prepared according to the amount required by the recipe, mixed and placed in a reverse flow extractor using ethanol and performed two extraction cycles with 70 % ethanol at 10 times the amount and with 70% ethanol at 8 times the amount, respectively, the duration of each cycle is 1 hour; filtered, removed the ethanol under reduced pressure, concentrated and dried to obtain a mixed extract of four medicinal materials of traditional Chinese medicine.

A mixed extract of four traditional Chinese medicine medicinal materials, consisting of Aurantii Fructus Immaturus, Trichosanthis Fructus, Cinnamomi Cortex, and red yeast rice, is mixed with Coptidis Rhizoma extract until homogeneous to obtain a composition based on extracts of traditional Chinese medicine medicinal materials of the present invention.

Content determination result (mg/g): total alkaloid content is 85.34 mg/g (including 62.24 mg/g berberine), total flavonoid content is 290.35 mg/g (including 170.37 mg /g of neohesperidin), and the content of lovastatin is 2.03 mg/g.

Example 3

Composition 3: 4.5 g (21.4 wt. %) Coptidis Rhizoma, 4.5 g (21.4 wt. %) Aurantii Fructus Immaturus, 0 g (0 wt. %) Pinelliae Rhizoma, 0 g (0 wt. %) Trichosanthis Fructus, 0 g (0 wt. %) Cinnamomi Cortex and 12 g (57.2 wt. %) red yeast rice, with a total daily dose of 21 g. The amount of extracts obtained is 5.21 g, the yield is 24.8%.

Method of preparation: an extract of the total alkaloids from Coptidis Rhizoma was obtained using the method of example 1.

A mixed extract of two medicinal materials of traditional Chinese medicine, consisting of bran-fried Aurantii Fructus Immaturus and red yeast rice, was prepared using the method of Example 1.

A mixed extract of two traditional Chinese medicine medicinal materials consisting of Aurantii Fructus Immaturus and red yeast rice was mixed with an extract of Coptidis Rhizoma until homogeneous to obtain a composition based on extracts of traditional Chinese medicine medicinal materials of the present invention.

Content determination result (mg/g): total alkaloid content is 43.51 mg/g (including 31.83 mg/g berberine), total flavonoid content is 234.38 mg/g (including 136.56 mg /g neohesperidin), and the content of lovastatin is 9.31 mg/g.

Example 4

Composition 4: 4.5 g (32.1 wt. %) Coptidis Rhizoma, 4.5 g (32.1 wt. %) Aurantii Fructus Immaturus, 0 g (0 wt. %) Pinelliae Rhizoma, 0 g (0 wt. %) Trichosanthis Fructus, 0.5 g (3.6 wt. %) Cinnamomi Cortex and 4.5 g (32.1 wt. %) red yeast rice, with a total daily administration dose of 14 g. is 3.74 g, with a yield of 26.7%.

Method of preparation: an extract of the total alkaloids from Coptidis Rhizoma was obtained using the method of example 1.

A mixed extract of three medicinal materials of traditional Chinese medicine, consisting of bran-fried Aurantii Fructus Immaturus, red yeast rice and Cinnamomi Cortex, was prepared using the method of Example 1.

A blended extract of three traditional Chinese medicine medicinal materials consisting of bran-fried Aurantii Fructus Immaturus, red yeast rice, and Cinnamomi Cortex was mixed with Coptidis Rhizoma extract until homogeneous to obtain a traditional Chinese medicine medicinal material extract composition of the present invention.

Content determination result (mg/g): total alkaloid content is 63.42 mg/g (including 46.25 mg/g berberine), total flavonoid content is 315.19 mg/g (including 185.75 mg /g neohesperidin), and the content of lovastatin is 4.63 mg/g.

Example 5

Composition 5: 6 g (33.8 wt. %) Coptidis Rhizoma, 1.5 g (8.5 wt. %) Aurantii Fructus Immaturus, 6 g (33.8 wt. %) Pinelliae Rhizoma, 0 g (0 wt. %) Trichosanthis Fructus, 1.25 g (7 wt %) Cinnamomi Cortex and 3 g (16.9 wt %) red yeast rice, for a total daily administration dose of 17.75 g. The amount of extracts obtained is 2 .64 g, with a yield of 14.9%.

Method of preparation: an extract of the total alkaloids from Coptidis Rhizoma was obtained using the method of example 1.

A mixed extract of four medicinal materials of traditional Chinese medicine, consisting of bran-fried Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice and Cinnamomi Cortex, was prepared using the method of Example 1.

A blended extract of four traditional Chinese medicine medicinal materials consisting of bran-fried Aurantii Fructus Immaturus, Pinelliae Rhizoma, red yeast rice, and Cinnamomi Cortex was mixed with Coptidis Rhizoma extract until homogeneous to obtain a traditional Chinese medicine medicinal material extract composition of the present invention. .

Content determination result (mg/g): total alkaloid content is 122.06 mg/g (including 89.03 mg/g berberine), total flavonoid content is 161.34 mg/g (including 92.86 mg /g of neohesperidin), and the content of lovastatin is 4.87 mg/g.

Example 6

Composition 6: 4.5 g (19.57 wt%) Coptidis Rhizoma, 4.5 g (19.56 wt%) Aurantii Fructus Immaturus, 4.5 g (19.57 wt%) Pinelliae Rhizoma, 6 g (26.09 wt%) Trichosanthis Fructus, 0.5 g (2.17 wt%) Cinnamomi Cortex and 3 g (13.04 wt%) red yeast rice, for a total daily dose of 23 g The amount of extracts obtained is 4.4 g, with a yield of 19.1%.

Method of preparation: an extract of the total alkaloids from Coptidis Rhizoma was obtained using the method of example 1.

90% of the total amount in the composition of red yeast rice, fried in the bran of Aurantii Fructus Immaturus, Pinelliae Rhizoma and Cinnamomi Cortex, was crushed to a coarse powder or cut medicinal raw materials in the form of pieces into pieces of a suitable size, received according to the required amount according to the recipe, mixed and placed in a reverse flow extractor using ethanol and performed two extraction cycles with 70% ethanol at 10 times the amount and with 70% ethanol at 8 times dilution, the duration of each cycle is 1 hour, the solution was settled and then filtered; in relation to the drug material Trichosanthis Fructus or its compressed pieces, two extraction cycles are carried out with 85% ethanol in a 7-fold amount and with 85% ethanol in a 5-fold amount, the duration of each cycle is 1 hour, the solution was settled and then filtered, the solution was combined with a solution of ethanol extracts of four traditional Chinese medicine medicinal materials, the ethanol was removed, concentrated under reduced pressure, dried, and ground to obtain a mixed extract of five traditional Chinese medicine medicinal materials.

Coptidis Rhizoma extract, a mixed extract of five medicinal materials of traditional Chinese medicine and 10% of the prescription amount of fine red yeast rice powder used as a disintegrant or tablet filler are homogeneously mixed to obtain a composition based on extracts of traditional Chinese medicine medicinal materials according to the present invention.

Content determination result (mg/g): total alkaloid content is 47.05 mg/g (including 31.58 mg/g berberine), total flavonoid content is 203.6 mg/g (including 108.3 mg /g neohesperidin), and the content of lovastatin is 2.59 mg/g.

Example 7

Composition 7: 4.5 g (19.57 wt%) Coptidis Rhizoma, 4.5 g (19.56 wt%) Aurantii Fructus Immaturus, 4.5 g (19.57 wt%) Pinelliae Rhizoma, 6 g (26.09 wt%) Trichosanthis Fructus, 0.5 g (2.17 wt%) Cinnamomi Cortex and 3 g (13.04 wt%) red yeast rice, for a total daily dose of 23 g The amount of extracts obtained is 5.62 g, with a yield of 24.4%.

Method of preparation: an extract of the total alkaloids from Coptidis Rhizoma was obtained using the method of example 1.

Bran-roasted Aurantii Fructus Immaturus was ground to a coarse powder or cut medicinal raw materials in pieces into pieces of suitable size, obtained according to the required amount according to the recipe and placed in a reverse flow extractor using ethanol, and two extraction cycles were performed with 70% ethanol in 8-fold quantity, the duration of each cycle is 1 hour; filtered, the ethanol removed under reduced pressure, and concentrated and dried to give an extract of the bran roasted Aurantii Fructus Immaturus.

Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine was ground to a coarse powder or cut medicinal raw materials in pieces into pieces of the appropriate size, obtained according to the required amount according to the recipe and placed in a reverse flow extractor using ethanol, and two extraction cycles were performed with 70% ethanol in 8-fold quantity, the duration of each cycle is 1 hour; filtered, ethanol removed under reduced pressure, and concentrated and dried to give an extract of Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine.

Whole Trichosanthis Fructus lump medicinal raw materials were obtained according to the required amount according to the prescription and placed in a reverse flow extractor using ethanol, and two extraction cycles were performed with 70% ethanol in 8 times the amount, the duration of each cycle is 1 hour; filtered, ethanol removed under reduced pressure, and concentrated and dried to give a whole Trichosanthis Fructus extract.

Red yeast rice was prepared according to the amount required by the recipe and placed in a reverse flow extractor using ethanol, and two extraction cycles were performed with 70% ethanol 8 times, the duration of each cycle was 1 hour; filtered, the ethanol removed under reduced pressure, and concentrated and dried to obtain a red yeast rice extract.

Cinnamomi Cortex was prepared according to the amount required by the recipe and placed in a reverse flow extractor using ethanol, and two extraction cycles were performed with 70% ethanol 8 times, the duration of each cycle was 1 hour; filtered, ethanol removed under reduced pressure, and concentrated and dried to give Cinnamomi Cortex extract.

Aurantii Fructus Immaturus, Cinnamomi Cortex, Pinelliae Rhizoma, Trichosanthis Fructus, and red yeast rice are mixed together uniformly to obtain a blended extract of traditional Chinese medicine five medicinal materials. The mixed extract of the five medicinal materials of traditional Chinese medicine is mixed with the extract of Coptidis Rhizoma until homogeneous, to obtain a composition based on extracts of traditional Chinese medicine medicinal materials of the present invention.

Content determination result (mg/g): total alkaloid content is 38.69 mg/g (including 28.10 mg/g berberine), total flavonoid content is 204.49 mg/g (including 123.2 mg /g of neohesperidin), and the content of lovastatin is 2.27 mg/g.

Comparative Example 1

Composition: 0 g (0 wt%) Coptidis Rhizoma, 4.5 g (24.32 wt%) Aurantii Fructus Immaturus, 4.5 g (24.32 wt%) Pinelliae Rhizoma, 6 g (32.43 wt.%) Trichosanthis Fructus, 0.5 g (2.70 wt.%) Cinnamomi Cortex and 3 g (16.21 wt.%) red yeast rice, with a total daily dose of administration of 18.5 g. extracts is 5.9 g, while the yield is 31.9%.

Preparation method: The preparation was carried out according to the extraction method of the corresponding ingredients from Example 6.

Content determination result (mg/g): total alkaloid content is 0 mg/g (including 0 mg/g berberine), total flavonoid content is 34.16 mg/g (including 20.11 mg/g neohesperidin) , and the content of lovastatin is 0.41 mg/g.

Comparative Example 2

Composition: 4.5 g (24.32 wt%) Coptidis Rhizoma, 0 g (0 wt%) Aurantii Fructus Immaturus, 4.5 g (24.32 wt%) Pinelliae Rhizoma, 6 g (32.43 wt.%) Trichosanthis Fructus, 0.5 g (2.70 wt.%) Cinnamomi Cortex and 3 g (16.21 wt.%) red yeast rice, with a total daily dose of administration of 18.5 g. extracts is 4.06 g, while the yield is 21.9%.

Preparation method: The preparation was carried out according to the extraction method of the corresponding ingredients from Example 6.

Content determination result (mg/g): total alkaloid content is 9.53 mg/g (including 6.98 mg/g berberine), total flavonoid content is 0 mg/g (including 0 mg/g neohesperidin) , and the content of lovastatin is 0.55 mg/g.

Comparative Example 3

Composition: 4.5 g (22.5 wt%) Coptidis Rhizoma, 4.5 g (22.5 wt%) Aurantii Fructus Immaturus, 4.5 g (22.5 wt%) Pinelliae Rhizoma, 6 g (30 wt. %) Trichosanthis Fructus, 0.5 g (2.5 wt. %) Cinnamomi Cortex and 0 g (0 wt. %) red yeast rice, with a total daily administration dose of 20 g. The amount of extracts obtained is 5.71 g, with a yield of 28.55%.

Preparation method: The preparation was carried out according to the extraction method of the corresponding ingredients from Example 6.

Content determination result (mg/g): total alkaloid content is 6.88 mg/g (including 4.96 mg/g berberine), total flavonoid content is 36.01 mg/g (including 21.67 mg /g neohesperidin), and the content of lovastatin is 0 mg/g.

Comparative Example 4

Composition: 4.5 g (50 wt%) Coptidis Rhizoma, 4.5 g (50 wt%) Aurantii Fructus Immaturus, 0 g (0 wt%) Pinelliae Rhizoma, 0 g (0 wt%) Trichosanthis Fructus , 0 g (0 wt.%) Cinnamomi Cortex, 0 g (0 wt.%) red yeast rice, with a total daily dose of administration of 9 g. The amount of extracts obtained is 2.66 g, while the yield is 9.56 %.

Preparation method: The preparation was carried out according to the extraction method of the corresponding ingredients from Example 6.

Content determination result (mg/g): total alkaloid content is 15.11 mg/g (including 10.65 mg/g berberine), total flavonoid content is 76.71 mg/g (including 46.62 mg /g neohesperidin), and the content of lovastatin is 0 mg/g.

Comparative Example 5

Composition: 4.5 g (29.03 wt%) Coptidis Rhizoma, 0 g (0 wt%) Aurantii Fructus Immaturus, 4.5 g (29.03 wt%) Pinelliae Rhizoma, 6 g (38.71 wt.%) Trichosanthis Fructus, 0.5 g (3.22 wt.%) Cinnamomi Cortex and 0 g (0 wt.%) red yeast rice, with a total daily dose of administration of 15.5 g. The amount of extracts obtained is 3.46 g, with a yield of 22.32%.

Preparation method: The preparation was carried out according to the extraction method of the corresponding ingredients from Example 6.

Content determination result (mg/g): total alkaloid content is 11.88 mg/g (including 8.12 mg/g berberine), total flavonoid content is 0 mg/g (including 0 mg/g neohesperidin) , and the content of lovastatin is 0 mg/g.

The formulations of drugs and the results of determining the content of active ingredients in examples 1-7 and comparative examples 1-5 are summarized below.

Table I. Drug formulations

Table II. Content determination results (mg/g)

Example 8

Composition 8: A mixture of extracts was prepared according to example 6.

Table III

Production method: the extract powder is weighed in a dose according to the composition and added to a dry granulator for granulation to obtain granules and sifting, magnesium stearate and silicon dioxide are added, the mixture is mixed until homogeneous and compressed into tablets using a special punch. Each tablet weighs 1.1 g and 1000 tablets are made. Compliant tablets have a moisture-resistant, reddish-brown film coating and a weight gain of 2-2.5%. The tablets were packaged to form the final product. Method of application: 2-5 tablets 2-3 times a day.

Table IV. Results of testing three batches of experimental products

Example 9

Composition 9: A mixture of extracts was prepared according to example 5.

Table V

The method of preparation is similar to that of example 8, with each tablet weighing 0.8 g.

Method of application: 1-2 tablets 3 times a day.

Table VI. Results of testing three batches of experimental products

Example 10

Composition 10: A mixture of extracts was prepared according to example 2.

Table VII

Producing Method: 50 g of calcium carbonate is added to 450 g of a mixture of extracts of six medicinal materials of traditional Chinese medicine, using 95% ethanol as a binder, a soft material is obtained, and the soft material is dried in an oven at a temperature of 40°C and formed into gelatin capsules to obtain only 1000 capsules.

Method of application: 2-3 capsules 3 times a day.

Table VIII. Results of testing three batches of experimental products

Пример 11

Example 11

Composition 11: A mixture of extracts was prepared according to example 7.

Table IX

Production method: a mixture of extracts was obtained according to the amount required by the recipe, and then mixed; the extract powder was weighed in an amount according to the composition, mannitol and aspartame were added, the mixture was stirred until homogeneous, and placed in a dry granulator to obtain granules and sieved, and the granules were packaged. Each sachet contains 4.1 g of granules and each sachet contains 2 g of extract.

How to use: 1-2 sachets 2-3 times a day.

Table X. Results of testing three batches of experimental products

I. Stability Study of the Traditional Chinese Medicine Medicinal Material Composition of the Present Invention

Sample Source: The traditional Chinese medicine medicinal material extract composition of the present invention is the extract obtained according to Example 8.

Stability studies have shown that the tablets with the composition of traditional Chinese medicine medicinal materials of the present invention easily absorb moisture and are more sensitive to high humidity than high temperature and light, and exposure to light has a significant effect on the content of lovastatin. In terms of final packaging for tablets, an inner packaging in the form of aluminum-plastic blisters is used, which are packed in an aluminum foil package. Conducted an accelerated test at a temperature of 40°C±2°C and a relative humidity of 75±5% for 6 months. The weight change rate due to moisture absorption of the tablets with the composition of traditional Chinese medicine medicinal materials of the present invention was in the range of 0.5%, which shows a good waterproofing and moisture-proofing effect of this package. The amounts of berberine hydrochloride, total alkaloids from Coptidis Rhizoma, total flavonoids and naringin from Aurantii Fructus Immaturus in the traditional Chinese medicine medicinal material composition tablet of the present invention change very little; the amount of lovastatin slightly changes, while the degree of change of the content is within 12.23%, and the change is mainly due to the interconversion of isomers and does not affect the efficacy.

Through stability studies of tablets with the composition of traditional Chinese medicine medicinal materials of the present invention, it was confirmed that an aluminum-plastic blister pack should be used as the inner package, which should be placed in an aluminum-plastic composite material bag; storage conditions should provide for sealed storage in a dry place; the shelf life of the drug is provisionally set at two years.

Table XI

Note: _ means no study is needed.

II. Study on the selection of the composition of the composition based on extracts of medicinal materials of traditional Chinese medicine according to the present invention

1. Laboratory animals: Wistar rats were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. License number: SCXK (Beijing) 2016-0011.

2. Testing method

2.1 Method for obtaining animal models with impaired glucose and lipid metabolism

2.1.1 High-fat diet and gastric tube loading with fat emulsion until lipid metabolism and insulin resistance develop

160 male Wistar rats (weighing 150-180 g) were purchased from Beijing Vital River Laboratory Animal Center. After adaptive feeding for a week, rats were randomly divided into a normal group and a high fat group. 20 rats from the normal group were fed a normal diet. 140 rats from the high fat group were fed a high fat diet and had their stomachs filled with a fat emulsion (1 ml/100 g weight/day) by gavage every day for 4 weeks.

After feeding for 4 weeks, rats from the normal group and rats from the high fat intake group took blood samples from the eye sockets to measure blood lipid levels. Compared to the normal weight group, serum triglycerides and cholesterol levels in rats from the high-fat group are significantly elevated, indicating impaired lipid metabolism.

After testing with the euglycemic clamp method, it was found that the glucose infusion rate (GIR) in the high fat group was significantly lower than in the normal group, indicating that the rats in the high fat group were significantly less sensitive to insulin, and the animals developed obvious symptoms of insulin resistance.

2.1.2 Intraperitoneal injection of small doses of STZ to produce glucose metabolism disorders

Preparation of 1% streptozocin (STZ) solution: add 0.1 mol/L sodium citrate buffer (pH 4.2) to exactly weighed 1.00 g streptozocin until the mixed solution reaches a volume of 100 ml and mix the mixed solution well to obtain 1% streptozocin solution (STZ). The solution is obtained immediately before use.

Animals in the high fat group did not receive food for 24 hours, while access to water was provided. 30 mg/kg/time of freshly prepared 1% solution of streptozocin in sodium citrate buffer was administered by intraperitoneal injection. Rats from the normal group were injected intraperitoneally with an equal volume of 0.1 mol/l sodium citrate buffer.

2.2 Blood lipid determination method

After 4 weeks of continuous feeding on a high fat diet, 2 rats were randomly selected from each cell of each group. 0.2 ml of blood was taken from the eye sockets of rats; were collected in EP tubes, after clotting, they were immediately placed in a centrifuge. Serum was collected and stored at -20°C. Serum was analyzed using the APPLYGEN reagent kit.

2.3 Euglycemic clamp method

After 4 weeks of continuous feeding with a high fat diet, one rat was randomly selected from each cell of the normal group (4 cells in total), while two rats were selected from the 4th cell; from the 4th, 9th, 14th, 19th and 24th cells of the high fat group (28 cells in total), one rat was randomly selected for the clamp study.

2.4 Method for preparing a prescription drug formulation

Production Methods According to the Examples: Compositions based on extracts of traditional Chinese medicine medicinal materials from Examples 1-7 were individually prepared according to the methods described in Examples 1-7. Preparation Method of Comparative Examples: Compositions based on extracts of traditional Chinese medicine medicinal materials from Comparative Examples 1 to 5 were prepared separately according to the method described in Example 6.

2.5 Route of administration

Model group and normal group: rats filled the stomach with 0.5 ml/100 g of body weight/day of saline using a gastric tube.

The dose of compositions based on extracts of traditional Chinese medicine medicinal materials from Examples 1 to 7 and Comparative Examples 1 to 5 for testing on rats was: 0.7 g/kg body weight/day. According to the equivalent dose calculation method, the rat dose is approximately equivalent to the next human dose of 0.11 g/kg body weight/day, which is equivalent to about 20-30 g of drug per day per person (based on a weight of 60 kg ). The dose for groups with other formulations was calculated in the same way.

Method for obtaining a test sample for filling the stomach with a gastric tube: 10.00 g of weighed sodium carboxymethyl cellulose was added to 1000 ml of pure water and swollen to obtain a 1% aqueous solution of CMC-Na. Weighed extracts with different compositions were dispersed in a 1% CMC-Na aqueous solution, and the mixed solution was stirred and mixed until uniform. The drug is taken once a week and stored in a refrigerator at 4°C. Introduction by filling the stomach with a gastric tube was carried out according to the following division into groups.

Table 1

На основе разделения по группам ежедневно с 8:30 до 10:30 осуществляли введение посредством наполнения желудка с помощью желудочного зонда, при этом доза составляла 0,5 мл/100 г веса/сутки. После 4 недель непрерывного введения подлежащие тестированию крысы не получали пищи ночью в течение 12 часов и с помощью глюкометра измеряли их уровень глюкозы в крови натощак (или измеряли с применением тест-полосок на уровень глюкозы в крови). Затем животных умерщвляли посредством пускания крови из бедренной артерии, отбирали образцы крови, отделяли сыворотку и измеряли соответственно пять показателей на уровень липидов в крови. В ходе испытания записывали изменение веса каждой крысы.

On the basis of division into groups daily from 8:30 to 10:30, administration was carried out by filling the stomach with a gastric tube, while the dose was 0.5 ml/100 g of weight/day. After 4 weeks of continuous administration, the test rats were fasted overnight for 12 hours and their fasting blood glucose levels were measured using a glucometer (or measured using blood glucose test strips). The animals were then euthanized by bleeding from the femoral artery, blood samples were taken, the serum was separated, and five blood lipid levels were measured respectively. During the test, the change in weight of each rat was recorded.

2.6 Method of statistical data analysis

All data were entered into an Excel spreadsheet to calculate the mean and standard deviation. Data for each group are presented as mean ± SD. SPSS 11.5 software was used to calculate data errors for each group. First, a homogeneity test of variance was performed, and then an analysis of variance was performed for each group.

3. Results of pharmacodynamic tests for examples

3.1 Results of blood lipid measurements after 4 weeks of feeding a high-fat diet

After feeding for 4 weeks in rats from the normal group and rats from the high fat group (high fat diet + fat emulsion), 2 rats were randomly selected from each cage and blood samples were taken from the eye area to measure blood lipids . The results are shown in Table 2.

Table 2. Blood lipid test results (mean ± SD)

(Compared to normal group ^*** p<0.001, ^** p<0.01, ^* p<0.05)

As shown in Table 2, after 4 weeks of feeding a high-fat diet and filling the stomach with a fat emulsion with a gastric tube, the blood lipid level in the high-fat group was significantly elevated, while being significantly different from the level in the normal group. This indicates that the lipid metabolism disorder model has been successfully developed.

3.2 Results of euglycemic clamp testing

Table 3 shows the results of testing using the euglycemic hyperinsulinemic clamp method.

Table 3. Results of testing each group using the euglycemic hyperinsulinemic clamp method (mean ± SD)

As shown in Table 3, after 4 weeks of feeding a high-fat diet and filling the stomach with a fat emulsion using a gastric tube, rats in the high-fat group significantly reduced the rate of glucose infusion and insulin sensitivity, and the animals clearly showed insulin resistance.

3.3 Results for grouping

Blood samples were taken from the tail vein 3 days after STZ administration to measure fasting blood glucose levels (after 12 hours of fasting). Rats with blood glucose levels above 26.6 mmol/l and below 13.3 mmol/l were excluded and selected and divided into groups of rats with blood glucose levels in the range of 13.3-26.6 mmol/l. Divided into groups of animals in accordance with the value of glucose in the blood, they were divided into 4 levels: 13.3-16.6 mmol/l, 16.7-19.9 mmol/l, 20.0-23.3 mmol/l and 23.4-26.6 mmol / l. Animals in each level were randomly divided into 8 groups according to their weight. The result for division into groups is shown in Table 4. The 8 groups consisted of composition group 1 to composition group 7 and a model group.

Table 4. Fasting blood glucose levels and weight of rats in each group (mean ± SD)

As shown in Table 4, the weight values of the animals were evenly distributed in the model group and different groups, and there is no significant difference in weight and blood glucose between the groups, indicating that the grouping was evenly distributed.

The control group consisted of 12 non-modeled normal rats weighing in the range of 358.75±12.08 g and with fasting blood glucose in the range of 4.94±0.34. There are significant differences for the control group compared to the post-model groups.

3.4 General condition of each group

Rats from the normal control group are in good condition, show fast reactions, normal locomotor ability, and have a shiny coat; rats from the model group after infusion of STZ are in a depressed state, show a slow response and slowness of movement. After 4 weeks of treatment, each treatment group showed improvement in overall condition to varying degrees.

3.5 Data on the change in blood glucose levels in the groups after treatment

The division into groups was carried out according to Table 4. After 4 weeks of continuous administration, the test rats were not fed at night for 12 hours, and their fasting blood glucose was measured (measurement using a glucometer and blood glucose test strips). The results are shown in Table 5.

Table 5. Fasting blood glucose values in each group after administration

(Compared to the model group ^*** p<0.001, ^** p<0.01, ^* p<0.05)

Note: in the division into groups according to Table 5, groups with composition 1-7 respectively represent examples 1-7 of the present invention, then by analogy.

As shown in Table 5, the fasting blood glucose level in formulation groups 3, 4, 6 and 7 was significantly lower compared to the level in the model group, with the strongest effect observed in formulation 6 group.

3.6 Data on the change in lipid levels in each group

After measuring the blood glucose level, the animals in each group were euthanized by femoral artery bleeding, blood samples were taken, serum was separated, and five blood lipid levels were measured respectively in the above manner. The results are shown in Table 6 and Table 7.

Table 6. Blood lipid values in each group after treatment (mean ± SD)

(Compared to the model group ^*** p<0.001, ^** p<0.01, ^* p<0.05)

Table 7. Blood lipid values in each group after treatment (mean ± SD)

(Compared to the model group ^*** p<0.001, ^** p<0.01, ^* p<0.05)

As shown in Table 6 and Table 7, the 5 blood lipid scores in each of the composition groups were significantly different from those in the model group. The level of triglycerides in each of the groups with compositions 5, 6 and 7 is significantly different from the level in the model group. The level of free cholesterol in each of the groups with compositions 1, 3, 4, 6 and 7 is different from the level in the model group. The level of total cholesterol in each of the groups with compositions 1, 3, 4, 5, 6 and 7 is different from the level in the model group. Analysis of the formulations and amounts in each formulation group showed that increasing the amount of red yeast rice was effective in lowering cholesterol but had no apparent effect on lowering triglycerides; in each group with the composition, there is a certain effect on lowering the level of triglycerides; presumably this is due to the amount and compatibility of Coptidis Rhizoma; in each group with the composition, there is a certain effect on increasing the level of high density lipoproteins; and in each group with the composition, there is a certain effect on the reduction of the level of low density lipoprotein, including groups with formulations 3 and 6, an effective decrease in the level of low density lipoprotein was noted.

The results of a comprehensive analysis of 5 indicators for the level of lipids in the blood showed that the most effective action in regulating the level of lipids in the blood was observed in group 6.

4. Results of pharmacodynamic tests in relation to comparative examples

4.1 Measurement Method Applied to Comparative Examples

Measurement of blood lipid levels after 4 weeks of feeding a high-fat diet, euglycemic clamp testing, analysis for division into groups, and analysis of the general condition for each group were performed using the methods indicated in sections 2.5, 2.6 and 3.1-3.4 .

4.2 Data on the change in blood glucose level in each group with a comparative example after treatment

After 4 weeks of continuous administration, using a method similar to that presented in section 3.5, the test rats were not fed at night for 12 hours and their fasting blood glucose levels were measured (measurement using a glucometer and blood glucose test strips). blood). The results are shown in Table 8.

Table 8. Fasting blood glucose values in each group

(Compared to the model group ^*** p<0.001, ^** p<0.01, ^* p<0.05)

As shown in Table 8, compared to the model group, fasting blood glucose is significantly lower in Example 6 (composition group 6) and in Comparative Examples 2 and 3, with the most significant decrease being in Example 6 (p<0.001) , and comparative example 1 showed the most mediocre glucose lowering effect, with the level closest to the value for the model group.

4.3. Data on the change in blood lipid levels in the comparative example groups after treatment

After measuring blood glucose levels using a method similar to that presented in section 3.6, animals in each group were sacrificed by bleeding from the femoral artery, blood samples were taken, serum was separated, and 5 indicators per blood lipid level were measured in the above manner. The results are shown in the following table.

Table 9. Blood lipid values in each group after treatment (µmol/l)

(Compared to the model group ^*** p<0.001, ^** p<0.01, ^* p<0.05)

As shown in Table 9, compared with the model group, the levels of triglycerides, free cholesterol, total cholesterol and low density lipoprotein of Example 6 are significantly reduced, and high density lipoprotein is significantly increased, and this effect is significantly superior to the effects of Comparative Examples 1 to 5. For Comparative Example 3, only triglycerides were effectively reduced, and for Comparative Examples 4 to 5, triglycerides, free cholesterol, total cholesterol, and LDL did not significantly decrease, and HDL was significantly reduced, showing the most mediocre effect.

A comprehensive analysis of the data from Table 8 and Table 9 shows that the glucose lowering effect for Comparative Examples 1 and 4-5 is relatively mediocre, the lipid lowering effect for Comparative Examples 3-5 is relatively mediocre, and the lipid and blood glucose lowering effect for Comparative Example 2 is worse than the effect of Example 6 of the present invention. As proved by Example 6 of the present invention, the traditional Chinese medicine extract composition of the present invention has excellent effects on both glucose lowering and blood lipid lowering.

III. Study of the pharmacodynamics of a composition based on extracts of medicinal materials of traditional Chinese medicine according to the present invention

Source of samples: composition based on extracts of traditional Chinese medicine medicinal materials of the present invention: extract obtained according to example 6.

Laboratory animals: KKAy mice provided by the Research Institute of Medical Laboratory Animals of the Academy of Medical Sciences of the People's Republic of China, C57BL/6J mice provided by Beijing Huafukang Biotechnology Co., LTD., SD rats provided by Beijing Vital River Laboratory Animal Technology Co., Ltd.

(1) Normal mice C57BL/6J

Female/male normal C57BL/6J mice, weight 18-20 g, were used respectively as normal controls for KKAy mice (female) with spontaneous type 2 diabetes mellitus and C57BL/6J mice (male) fed a high fat diet .

(2) KKAy mice with spontaneous type 2 diabetes mellitus

Female KKAy mice with spontaneous type 2 diabetes mellitus were fed a special diet designed for KKAy mice, 5 mice per cage. After the mouse weight exceeded 40 g, predictions were made for a number of indicators, including blood glucose, blood lipids, and insulin resistance. Depending on the level of fasting blood glucose, the percentage decrease in blood glucose after 40 minutes. after testing for insulin tolerance, blood triglycerides (TG), total cholesterol (TC) and weight, the mice were divided into 5 groups of 12 mice in each group.

(3) C57BL/6J mice fed a high fat diet

Male C57BL/6J mice were fed a special high fat diet with 5 animals per cage. After the mouse weight exceeded 45 g, predictions were made for a number of indicators, including blood glucose, blood lipids, and insulin resistance. Depending on the level of fasting blood glucose, the percentage decrease in blood glucose after 40 minutes. after insulin tolerance test, blood TG level, blood TC level and weight, the mice were divided into 5 groups of 12 mice in each group.

Test 1 Effect of Traditional Chinese Medicine Material Extract Composition of the Present Invention on Insulin Tolerance in KKAy Mice with Spontaneous Type 2 Diabetes Mellitus

KKAy mice with spontaneous type 2 diabetes mellitus were divided into 5 groups based on the following four measures: percent reduction in blood glucose after 40 min. after testing for insulin tolerance, fasting blood glucose, blood TG, and blood TC. The groups were respectively a model control group (Con, filling the stomach with water of an equivalent volume using a gastric tube), a positive control group (Met, filling the stomach with 150 mg/kg body weight of metformin using a gastric tube), and three groups of dosing a composition based on an extract of medicinal traditional Chinese medicine of the present invention. Animals in the three dose groups were subjected to gastric filling with 500, 1000 and 1500 mg/kg of the composition based on the extract of drugs of traditional Chinese medicine using a gastric tube; and the content of active ingredients for each dosing group is shown in Table 10.

Table 10 Content of Active Ingredients in Different Dose Groups of Composition Based on Traditional Chinese Medicine Extract (g/kg)

Female C57BL/6J mice were used as a control group with normal animals (Nor, filling the stomach with water of an equivalent volume using a gastric tube, n=10). Filling the stomach with a gastric tube was carried out once a day continuously for 58 days, on the 14th day of administration, the mice were tested for insulin tolerance. On the day of testing, the animals did not receive food for 4 hours (with access to water), blood was taken from the tip of the tail (0 minutes), and then a subcutaneous injection of regular insulin (0.4 U/kg) was performed and blood was resampled was carried out 40 and 90 minutes after injection. The blood glucose level was determined using the glucose oxidase method (hereinafter by analogy), and the area under the blood glucose curve (AUC) was calculated.

The results demonstrated (in Table 11 and Table 12) that the blood glucose level in the model control group (Con) did not significantly decrease 40 minutes after the subcutaneous injection of insulin (0.4 U/kg), indicating the presence of clear signs of insulin resistance. in KKAy mice with spontaneous type 2 diabetes mellitus. In contrast to the Con group, the blood glucose level of mice in the three dose groups using 500, 1000 and 1500 mg/kg of the composition based on the extracts of traditional Chinese medicine medicinal materials of the present invention decreased at 0 minutes and 40 minutes after subcutaneous injection of insulin. ; and the area under the blood glucose curve also decreased by 7.8%, 26.1% and 41.8% respectively, respectively, with a certain dose-dependence observed. This indicates that the composition based on extracts of medicinal materials of traditional Chinese medicine is able to improve the state of insulin resistance in KKAy mice with spontaneous type 2 diabetes mellitus to a certain extent.

Table 11 Effect of Traditional Chinese Medicine Material Composition of the Invention on Insulin Tolerance in Spontaneous Type 2 Diabetes KKAy Mice

(Compared with the model group, n=12, ^* p<0.05, ^** p<0.01, ^*** p<0.001)

Table 12 Effect of the Traditional Chinese Medicine Medicinal Material Composition of the Present Invention on the Area Under the Curve of Blood Sugar in Insulin Tolerance (AUC) in KKAy Mice

(Compared with the model group, n=12, ^* p<0.05, ^** p<0.01, ^*** p<0.001)

Test 2: Effect of the Traditional Chinese Medicine Material Extract Composition of the Present Invention on Oral Glucose Tolerance in Spontaneous Type 2 Diabetes KKAy Mice.

KKAy mice with spontaneous type 2 diabetes mellitus were divided into 5 groups based on the following five measures: percent reduction in blood glucose after 40 min. after insulin tolerance test, fasting blood glucose, blood TG, blood TC, and weight. The groups were respectively a model control group (Con, filling the stomach with water of an equivalent volume using a gastric tube), a positive control group (Met, filling the stomach with 150 mg/kg body weight of metformin using a gastric tube), and three groups of dosing a composition based on an extract of medicinal traditional Chinese medicine of the present invention. Animals in the three dose groups were subjected to gastric filling with 500, 1000 and 1500 mg/kg of the composition based on the extract of drugs of traditional Chinese medicine using a gastric tube. Female C57BL/6J mice were used as a control group with normal animals (Nor, filling the stomach with water of an equivalent volume using a gastric tube, n=10). Filling the stomach with a gastric tube was carried out once a day continuously for 58 days, on the 28th day of administration, the animals in each group were subjected to oral testing for glucose tolerance. On the day of testing, animals in each group did not receive food for 4 hours (with access to water), blood was taken from the tip of the tail (0 minutes), then the stomach was filled with glucose solution (2.0 g/kg) using a gastric probe and blood sampling was carried out 30, 60 and 120 minutes after the carbohydrate load. The blood glucose level was measured and the area under the blood glucose curve (AUC) was calculated.

The results showed (Table 13 and Table 14) that, compared with the normal control group (Nor), the model control group (Con) showed a significant increase in blood glucose after filling the stomach with glucose solution using a gastric tube, indicating an obvious violation glucose tolerance in KKAy mice with spontaneous type 2 diabetes mellitus. In contrast to the Con group, spontaneous type 2 diabetic KKAy mice in the three dose groups of 500, 1000, and 1500 mg/kg of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention, when tested after oral carbohydrate loading, a decrease in blood glucose to a certain extent, and the area under the blood glucose curve decreased by 12%, 12.5% and 17.2%, respectively, while the effect of lowering blood glucose in the dose group of 1500 mg / d has a statistically significant difference. This indicates that the composition based on extracts of traditional Chinese medicine medicinal materials can improve glucose intolerance to a certain extent in KKAy mice with spontaneous type 2 diabetes mellitus.

Table 13 Effect of Traditional Chinese Medicine Medicinal Material Composition of the Present Invention on Oral Glucose Tolerance in KKAy Mice with Spontaneous Type 2 Diabetes Mellitus

(Compared with the model group, n=10-12, ^* p<0.05, ^** p<0.01, ^*** p<0.001)

Table 14. Effect of the Traditional Chinese Medicine Medicinal Material Composition of the Present Invention on the Area Under the Blood Glucose Curve for Glucose Tolerance (AUC) in KKAy Mice

(Compared to the model group, n=10-12, ^* p<0.05, ^** p<0.01)

Test 3 Effect of Traditional Chinese Medicine Material Extract Composition of the Present Invention on Fasting Blood Glucose Level, Blood Insulin Level and Insulin Sensitivity Index in KKAy Mice with Spontaneous Type 2 Diabetes Mellitus

KKAy mice with spontaneous type 2 diabetes mellitus were divided into 5 groups based on the following five measures: percent reduction in blood glucose after 40 min. after insulin tolerance test, fasting blood glucose, blood TG, blood TC, and weight. The groups were respectively a model control group (Con, filling the stomach with water of an equivalent volume using a gastric tube), a positive control group (Met, filling the stomach with 150 mg/kg body weight of metformin using a gastric tube), and three groups of dosing a composition based on an extract of medicinal traditional Chinese medicine of the present invention. Animals in the three dose groups were subjected to gastric filling with 500, 1000 and 1500 mg/kg of the composition based on the extract of drugs of traditional Chinese medicine using a gastric tube. Female C57BL/6J mice were used as a control group with normal animals (Nor, filling the stomach with water of an equivalent volume using a gastric tube, n=10). Filling the stomach with a gastric tube was carried out once a day continuously for 58 days; on the 28th day of administration, simultaneously with testing for glucose tolerance in animals in each group, fasting blood glucose and blood insulin levels were measured (by ELISA method), and insulin sensitivity index (ISI) was calculated, while ISI = 1/ (FBG _mg/dL X FBI _ng/mL ).

The results showed (Table 15) that, compared with the Con group, in the 500 mg/kg dose group and the 1500 mg/kg dose group, the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention can achieve significant reduction in fasting blood glucose, blood insulin levels, and insulin sensitivity index (ISI) in KKAy mice with spontaneous type 2 diabetes mellitus.

Table 15 Effect of Traditional Chinese Medicine Medicinal Material Composition of the Present Invention on Fasting Blood Glucose Level, Blood Insulin Level and Insulin Sensitivity Index in KKAy Mice

(Compared with the model group, n=10-12, ^* p<0.05, ^** p<0.01, ^*** p<0.001)

Test 4 Effect of Prolonged Administration of the Composition Based on Extracts of Traditional Chinese Medicine Medicinal Materials of the Present Invention on Fasting Blood Glucose in KKAy Mice with Spontaneous Type 2 Diabetes Mellitus

KKAy mice with spontaneous type 2 diabetes mellitus were divided into 5 groups based on the following five measures: percent reduction in blood glucose after 40 min. after insulin tolerance test, fasting blood glucose, blood TG, blood TC, and weight. The groups were respectively a model control group (Con, filling the stomach with water of an equivalent volume using a gastric tube), a positive control group (Met, filling the stomach with 150 mg/kg body weight of metformin using a gastric tube), and three groups of dosing a composition based on an extract of medicinal traditional Chinese medicine of the present invention. Animals in the three dose groups were subjected to gastric filling with 500, 1000 and 1500 mg/kg of the composition based on the extract of drugs of traditional Chinese medicine using a gastric tube. Female C57BL/6J mice were used as a control group with normal animals (Nor, filling the stomach with water of an equivalent volume using a gastric tube, n=10). Filling the stomach with a gastric tube was carried out once a day continuously for 58 days; animals in each group did not receive food for 4 hours (with access to water), on the 14th, 21st, 28th and 40th day of administration, blood was taken from the tip of the tail with the measurement of fasting blood glucose .

The results showed (Table 16) that, compared with the Con group, the three dose groups of 500, 1000, and 1500 mg/kg of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention can achieve different degrees of reduction in glucose levels. in the fasting blood of KKAy mice with spontaneous type 2 diabetes mellitus, while in the 1500 mg/kg dose group, the percentage decrease in blood glucose in KKAy mice with spontaneous type 2 diabetes at different time points was respectively 43.9% , 23.6%, 29.8% and 22.4%. This indicates that the long-term administration of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention can provide a clear control of blood glucose levels in KKAy mice with spontaneous type 2 diabetes mellitus.

Table 16. Effect of prolonged administration of the traditional Chinese medicine medicinal material composition of the present invention on fasting blood glucose in KKAy mice with spontaneous type 2 diabetes mellitus

(Compared with the model group, n=10-12, ^* p<0.05, ^** p<0.01, ^*** p<0.001)

Test 5 Effect of Traditional Chinese Medicine Material Extract Composition of the Present Invention on Glycated Hemoglobin (HbA1c) Level in KKAy Mice with Spontaneous Type 2 Diabetes Mellitus

KKAy mice with spontaneous type 2 diabetes mellitus were divided into 5 groups based on the following five measures: percent reduction in blood glucose after 40 min. after insulin tolerance test, fasting blood glucose, blood TG, blood TC, and weight. The groups were respectively a model control group (Con, filling the stomach with water of an equivalent volume using a gastric tube), a positive control group (Met, filling the stomach with 150 mg/kg body weight of metformin using a gastric tube), and three groups of dosing a composition based on an extract of medicinal traditional Chinese medicine of the present invention. Animals in the three dose groups were subjected to gastric filling with 500, 1000 and 1500 mg/kg of the composition based on the extract of drugs of traditional Chinese medicine using a gastric tube. Female C57BL/6J mice were used as a control group with normal animals (Nor, filling the stomach with water of an equivalent volume using a gastric tube, n=10). Filling the stomach with a gastric tube was carried out once a day continuously for 58 days; on the 48th day of administration, blood was taken from the tip of the tail and the level of glycated hemoglobin was measured.

The results demonstrated (Table 17) that, compared with the control group with normal animals in the model control group, KKAy mice with spontaneous type 2 diabetes showed a significant increase in the level of HbA1c. Long-term administration of the composition based on the extracts of medicinal materials of traditional Chinese medicine according to the present invention (500, 1000 and 1500 mg/kg) is able to reduce the level of HbA1c in KKAy mice with spontaneous type 2 diabetes mellitus by 0.77, 0.87 and 1.03, respectively. , there are significant differences between the medium and high dose groups. This indicates that the long-term administration of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention can provide a clear control of fluctuating blood glucose levels in KKAy mice with spontaneous type 2 diabetes mellitus.

Table 17. Effect of the traditional Chinese medicine composition of the present invention on the level of glycated hemoglobin in KKAy mice with spontaneous type 2 diabetes mellitus (day 48)

(Compared with the model group, n=11-12, ^* p<0.05, ^*** p<0.001)

Test 6 Effect of Traditional Chinese Medicine Material Extract Composition of the Present Invention on Blood Triglycerides (TG) and Total Cholesterol (TC) in KKAy Mice

KKAy mice with spontaneous type 2 diabetes mellitus were divided into 5 groups based on the following five measures: percent reduction in blood glucose after 40 min. after insulin tolerance test, fasting blood glucose, blood TG, blood TC, and weight. The groups were respectively a model control group (Con, filling the stomach with water of an equivalent volume using a gastric tube), a positive control group (Met, filling the stomach with 150 mg/kg body weight of metformin using a gastric tube), and three groups of dosing a composition based on an extract of medicinal traditional Chinese medicine of the present invention. Animals in the three dose groups were subjected to gastric filling with 500, 1000 and 1500 mg/kg of the composition based on the extract of drugs of traditional Chinese medicine using a gastric tube. Female C57BL/6J mice were used as a control group with normal animals (Nor, filling the stomach with water of an equivalent volume using a gastric tube, n=10). Filling the stomach with was carried out once a day continuously for 58 days; at the 14th injection, simultaneously with testing for insulin tolerance in mice, blood was taken to measure the level of triglycerides (TG) and total cholesterol (TC).

The results showed (Table 18) that, compared with the control group (Con), in the 1500mg/kg dose group, the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention can achieve a significant reduction in serum TG levels in mice. KKAy with spontaneous type 2 diabetes mellitus, and at the same time, there was no significant effect on the level of TC in the blood serum.

Table 18 Effect of the Traditional Chinese Medicine Medicinal Material Composition of the Present Invention on Blood TG Level and Blood TC Level in Spontaneous Type 2 Diabetes KKAy Mice

(Compared with the model group, n=10-12, ^* p<0.05, ^*** p<0.001)

Test 7 Effect of long-term administration of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention on insulin resistance in KKAy mice with spontaneous diabetes mellitus 2 (euglycemic hyperinsulinemic clamp method)

KKAy mice with spontaneous type 2 diabetes mellitus were divided into 5 groups based on the following five measures: percent reduction in blood glucose after 40 min. after insulin tolerance test, fasting blood glucose, blood TG, blood TC, and weight. The groups were respectively a model control group (Con, filling the stomach with water of an equivalent volume using a gastric tube), a positive control group (Met, filling the stomach with 150 mg/kg body weight of metformin using a gastric tube), and three groups of dosing a composition based on an extract of medicinal traditional Chinese medicine of the present invention. Animals in the three dose groups were subjected to gastric filling with 500, 1000 and 1500 mg/kg of the composition based on the extract of drugs of traditional Chinese medicine using a gastric tube. Female C57BL/6J mice were used as a control group with normal animals (Nor, filling the stomach with water of an equivalent volume using a gastric tube, n=10). Filling the stomach with a gastric tube was carried out once a day continuously for 58 days; after 54 days of administration to animals in each group, 5-6 mice from different groups were randomly selected for daily euglycemic hyperinsulinemic clamp testing.

The results showed (Table 19) that, compared with the control group with normal animals (Nor), when the steady state blood glucose concentration (6.0 mmol/L) was reached under high insulin conditions, KKAy mice with spontaneous diabetes mellitus 2 type in the model control group (Con) there is a significant decrease in glucose infusion rate (GIR), which indicates the presence of a clear insulin resistance in KKAy mice with spontaneous type 2 diabetes mellitus. In contrast to the model control group (Con) in the three dose groups of 500, 1000 and 1500 mg/kg of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention, it was found that prolonged administration can increase the GIR value in KKAy mice. with spontaneous type 2 diabetes by 12.3%, 54.3% and 70.4%, respectively. This indicates that the long-term administration of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention is able to improve insulin sensitivity in KKAy mice with spontaneous type 2 diabetes mellitus.

Table 19. Effect of prolonged administration of the traditional Chinese medicine medicinal material composition of the present invention on the rate of glucose infusion in KKAy mice with spontaneous type 2 diabetes mellitus

(Compared with the model group, n=3-5, ^* p<0.05, ^** p<0.01, ^*** p<0.001)

Test 8 Effect of Traditional Chinese Medicine Material Extract Composition of the Present Invention on Blood Pressure in KKAy Mice with Spontaneous Type 2 Diabetes Mellitus

KKAy mice with spontaneous type 2 diabetes mellitus were divided into 5 groups based on the following five measures: percent reduction in blood glucose after 40 min. after insulin tolerance test, fasting blood glucose, blood TG, blood TC, and weight. The groups were respectively a model control group (Con, filling the stomach with water of an equivalent volume using a gastric tube), a positive control group (Met, filling the stomach with 150 mg/kg body weight of metformin using a gastric tube), and three groups of dosing a composition based on an extract of medicinal traditional Chinese medicine of the present invention. Animals in the three dose groups were subjected to gastric filling with 500, 1000 and 1500 mg/kg of the composition based on the extract of drugs of traditional Chinese medicine using a gastric tube. Female C57BL/6J mice were used as a control group with normal animals (Nor, filling the stomach with water of an equivalent volume using a gastric tube, n=10). Filling the stomach with a gastric tube was carried out once a day continuously for 58 days, on the 53rd day of administration, the animals in each group were measured using an intelligent non-invasive tonometer (BP98A, Beijing Ruanlong Biotechnology Co., Ltd.).

The results demonstrated (Table 20) that the continuous administration of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention (500, 1000 and 1500 mg/kg) for 53 days can significantly reduce blood pressure in KKAy mice with spontaneous sugar type 2 diabetes (note: due to their small weight and small tail, it was not possible to measure blood pressure with a non-invasive blood pressure monitor in normal control mice).

Table 20 Effect of Traditional Chinese Medicine Medicinal Material Composition of the Present Invention on Blood Pressure in KKAy Mice with Spontaneous Type 2 Diabetes Mellitus

(Compared with the model group, n=11-12, ^* p<0.05, ^** p<0.01)

Test 9 Effect of the traditional Chinese medicine extract composition of the present invention on insulin tolerance in C57BL/6J mice fed a high fat diet

C57BL/6J mice fed a high-fat diet and weighing over 45 g were selected for pre-testing for measures including blood glucose, lipid levels, and insulin resistance, and divided into 5 groups based on the following five measures: percent decrease in blood glucose levels after 40 minutes. after insulin tolerance test, fasting blood glucose level, blood TG level, TC level and weight, with 12 mice in each group. The groups were respectively a model control group (Con), a positive control group with metformin (Met, 150 mg/kg), and three dose groups (500, 1000 and 1500 mg/kg) using a composition based on an extract of traditional Chinese medicine drugs. according to the present invention. Male C57BL/6J mice were used as a control group with normal animals. Animals in the positive control group and each test drug group were subjected to daily gastric filling with the appropriate dose of the test drug using a gastric tube. The normal control group and the model control group were subjected to filling the stomach with an equivalent volume of water using a gastric tube once a day continuously for 30 days. On the 10th day after administration, the animals in each group were tested for insulin tolerance. On the day of testing, the animals did not receive food for 4 hours (with access to water), blood was taken from the tip of the tail (0 minutes), and then a subcutaneous injection of regular insulin (0.4 U/kg) was performed. Blood sampling was carried out after 40 min. and 90 min. after injection with blood glucose measurement and AUC calculation.

The results showed (Table 21 and Table 22) that when comparing blood glucose measurement 40 minutes after insulin injection and blood glucose measurement at 0 minutes, normal control group C57BL/6J mice showed a decrease in blood glucose after 40 min. after insulin injection by 40.3%, and C57BL/6J mice fed a high-fat diet from the model control group showed a decrease in blood glucose levels after 40 minutes. after insulin injection by 26.7%; indicating the presence of insulin resistance in obese C57BL/6J mice.

In contrast to the model control group (Con), the 500mg/kg dose group and the 1500mg/kg dose group of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention showed a significant decrease in blood glucose and AUC in C57BL/6J mice fed a high fat diet at 40 min. and 90 min. after insulin injection, and in the 1000 mg/kg dose group, there was a significant decrease in blood glucose and AUC in C57BL/6J mice fed a high fat diet after 90 minutes. after insulin injection. This indicates that the composition based on extracts of medicinal materials of traditional Chinese medicine is able to significantly improve the state of insulin resistance in C57BL/6J mice fed a high fat diet.

Table 21 Effect of the traditional Chinese medicine composition of the present invention on insulin tolerance in C57BL/6J mice fed a high fat diet

(Compared with the model group, n=10-12, ^* p<0.05, ^** p<0.01, ^*** p<0.001)

Table 22 Effect of the traditional Chinese medicine composition of the present invention on insulin tolerance by AUC in C57BL/6J mice fed a high fat diet

(Compared to the model group, n=10-12, ^* p<0.05, ^** p<0.01)

Test 10 Effect of the traditional Chinese medicine extract composition of the present invention on oral glucose tolerance in C57BL/6J mice fed a high fat diet

C57BL/6J mice fed a high-fat diet and weighing over 45 g were selected for pre-testing for measures including blood glucose, lipid levels, and insulin resistance, and divided into 5 groups based on the following five measures: percent decrease in blood glucose levels after 40 minutes. after insulin tolerance test, fasting blood glucose level, blood TG level, TC level and weight, with 12 mice in each group. The groups were respectively a model control group (Con), a positive control group with metformin (Met, 150 mg/kg), and three dose groups (500, 1000 and 1500 mg/kg) using a composition based on an extract of traditional Chinese medicine drugs. according to the present invention. Male C57BL/6J mice were used as a control group with normal animals. Animals in the positive control group and each test drug group were subjected to daily gastric filling with the appropriate dose of the test drug using a gastric tube. The normal control group and the model control group were subjected to filling the stomach with an equivalent volume of water using a gastric tube once a day continuously for 30 days. On the 29th day after administration, the animals in each group were tested for glucose tolerance. On the day of testing, the animals did not receive food for 4 hours (with access to water), blood was taken from the tip of the tail (0 minutes), and then the stomach was filled with glucose solution (2.0 g/kg) using a gastric tube. Blood sampling was carried out after 30 minutes, 60 minutes. and 120 min. after a carbohydrate load with blood glucose measurement and AUC calculation.

The results showed (Table 23 and Table 24) that in the 500 mg/kg dose and 1000 mg/kg dose groups of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention, there was a significant decrease in glucose levels in blood samples from C57BL/6J mice fed a high-fat diet at 30, 60, and 120 minutes after an oral carbohydrate load; in the 1500 mg/kg dose group, there was a significant decrease in blood glucose levels 30 minutes after oral carbohydrate loading; and the three dose groups showed reductions in AUC of 10.5%, 14.8%, and 1.7%, respectively. This indicates that the traditional Chinese medicine extract composition of the present invention is able to improve oral glucose tolerance abnormalities in C57BL/6J mice fed a high fat diet.

Table 23. Effect of the traditional Chinese medicine composition of the present invention on oral glucose tolerance in C57BL/6J mice fed a high fat diet

(Compared with the model group, n=10-12, ^* p<0.05, ^** p<0.01, ^*** p<0.001)

Table 24 Effect of the traditional Chinese medicine composition of the present invention on oral glucose tolerance AUC in C57BL/6J mice fed a high fat diet

(Compared with the model group, n=10-12, ^** p<0.01, ^*** p<0.001)

In order to investigate the effect of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention on the signs of metabolic syndrome, such as insulin resistance, glucose and lipid metabolism, blood pressure and weight, a pharmacodynamic test was carried out in which KKAy mice were used as a laboratory animal model with spontaneous type 2 diabetes mellitus and insulin resistance and obesity and C57BL/6J mice fed a high fat diet.

The results of the pharmacodynamic test showed that the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention has the following effects: (1) lowering blood glucose; (2) decrease in the level of insulin in the blood; (3) improvement in glucose tolerance as measured by an oral test; (4) decrease in the level of glycated hemoglobin (HbA1c); (5) improved insulin sensitivity index and improvement in insulin resistance; (6) lowering blood triglycerides and trending lower total cholesterol; and (7) lowering blood pressure.

VI. Safety of the Composition Based on the Extracts of Medicinal Materials of Traditional Chinese Medicine of the Present Invention

Laboratory Animals: C57BL/6J mice provided by Beijing Huafukang Biotechnology Co., LTD. and Beagle dogs provided by Beijing Maas Biotechnology Co., LTD.

In an acute toxicity test on mice that were orally administered the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention, the LD ₅₀ value was measured to be more than 20,000 mg/kg.

In an acute toxicity test on beagle dogs that were orally administered the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention, the lethal dose was measured to be more than 7500 mg/kg, and the main toxic reaction was vomiting after administration. The 20,000 mg/kg dose is 200 times the suggested clinical dose, and the 7500 mg/kg dose is 75 times the suggested clinical dose.

The results of long-term toxicity testing in rats that were orally administered the composition based on the extracts of traditional Chinese medicine medicinal materials of the present invention showed that it can cause reversible gastric damage in rats such as proventricular hyperkeratosis and parakeratosis and destruction of the mucous membrane; as well as liver damage such as reversible hepatocellular hypertrophy and refractory bile duct hyperplasia. Long-term toxicity testing used doses that were 8, 28, and 98 times the intended clinical dose; and thus established the relationship between the toxic effect and long-term use of the test product in high doses and the solvent used to obtain the test product. Thus, the expected clinical dose of the composition based on extracts of traditional Chinese medicine medicinal materials of the present invention is safe.

The foregoing description is solely a description of specific embodiments of the present invention, however, the protection scope of the present invention is not limited to this description; any modifications or substitutions that any person skilled in the art may suggest within the technical field disclosed by the present invention are within the protection scope of the present invention. Therefore, the protection scope of the present invention is determined by the protection scope of the claims.

Applicability

Presented in accordance with the embodiments of the present invention, the composition based on medicinal materials of traditional Chinese medicine and the composition based on extracts of medicinal materials of traditional Chinese medicine are applicable in the technical field of pharmaceuticals and are especially applicable for the treatment of metabolic syndrome.

Claims (18)

1. A traditional Chinese medicine composition for treating metabolic syndrome, wherein the traditional Chinese medicine composition contains Coptidis Rhizoma, Aurantii Fructus Immaturus, Pinelliae Rhizoma, Trichosanthis Fructus, Cinnamomi Cortex, and red yeast rice as raw materials, wherein the composition contains 19.57 wt. % Coptidis Rhizoma, 19.56 wt. % Aurantii Fructus Immaturus, 19.57 wt. % Pinelliae Rhizoma, 26.09 wt. % Trichosanthis Fructus, 2.17 wt. % Cinnamomi Cortex and 13.04 wt. % red yeast rice. 2. Traditional Chinese medicine composition according to claim 1, wherein Aurantii Fructus Immaturus is Aurantii Fructus Immaturus roasted in bran, Pinelliae Rhizoma is Pinelliae Rhizoma Praeparatumcum Zingibere Et Alumine, and Trichosanthis Fructus is Trichosanthis Fructus whole. 3. A composition based on medicinal materials of traditional Chinese medicine for the treatment of metabolic syndrome, where the composition contains Coptidis Rhizoma, Aurantii Fructus Immaturus, red yeast rice and optionally one or more components selected from the group consisting of Pinelliae Rhizoma, Trichosanthis Fructus and Cinnamomi Cortex in form of raw material, where the composition contains: 7.9-45 wt. % Coptidis Rhizoma, 8.5-32.1 wt. % Aurantii Fructus Immaturus, 0-33.8 wt. % Pinelliae Rhizoma, 0-26.09 wt. % Trichosanthis Fructus, 0-13.1 wt. % Cinnamomi Cortex and 13.04-57.2 wt. % red yeast rice. 4. Pharmaceutical composition for the treatment of metabolic syndrome, containing a composition based on medicinal materials of traditional Chinese medicine according to any one of paragraphs. 1-3 and a pharmaceutically acceptable carrier. 5. A method of obtaining a composition for the treatment of metabolic syndrome from the composition according to any one of paragraphs. 1-3, where (a-1) Coptidis Rhizoma is extracted using an aqueous acid solution and filtered, concentrated, subjected to alkaline precipitation, salting out, filtered, dried and milled to obtain a total alkaloid extract from Coptidis Rhizoma; (a-2) four drug materials, including Red Yeast Rice, 90% of the prescription amount, Aurantii Fructus Immaturus, Pinelliae Rhizoma, and Cinnamomi Cortex, are extracted with 70% ethanol and filtered; (a-3) Trichosanthis Fructus is extracted using 85% ethanol, the Trichosanthis Fructus extract solution is combined with the extract solution of four medicinal materials of traditional Chinese medicine, the ethanol is removed under reduced pressure, concentrated, dried, and pulverized to obtain a mixed extract of five medicinal materials of traditional Chinese medicine consisting of Aurantii Fructus Immaturus, Pinelliae Rhizoma, Red Yeast Rice, Cinnamomi Cortex and Trichosanthis Fructus; and (a-4) Coptidis Rhizoma total alkaloid extract, mixed extract of five medicinal materials of traditional Chinese medicine, and fine powder of red yeast rice, accounting for 10% of the prescription amount, are homogeneously mixed to obtain a composition based on medicinal materials of traditional Chinese medicine . 6. Pharmaceutical preparation for the treatment of metabolic syndrome, containing a composition based on medicinal materials of traditional Chinese medicine according to any one of paragraphs. 1-3, where the pharmaceutical preparation provides injection solutions, tablets, capsules, granules and solutions for oral administration. 7. The use of compositions based on medicinal materials of traditional Chinese medicine according to any one of paragraphs. 1-3 for the treatment of a disease selected from: a. metabolic syndrome; b. disorders of blood glucose and lipid metabolism at an early stage of the metabolic syndrome. 8. The use of compositions based on medicinal materials of traditional Chinese medicine according to any one of paragraphs. 1-3 to provide one or more of the following effects: lower blood glucose, lower blood insulin, improve oral glucose tolerance, lower glycated hemoglobin (HbAlc), improve insulin sensitivity index, improve insulin resistance, lowering blood triglycerides, lowering total cholesterol and lowering blood pressure. 9. The use of a pharmaceutical preparation according to claim 6 for the treatment of a disease selected from: a. metabolic syndrome; b. disorders of blood glucose and lipid metabolism at an early stage of the metabolic syndrome. 10. The use of a pharmaceutical preparation according to claim 6 to provide one or more of the following effects: lowering blood glucose levels, lowering blood insulin levels, improving oral glucose tolerance, lowering glycated hemoglobin (HbAlc), increasing sensitivity index to insulin, improving insulin resistance, lowering blood triglycerides, lowering total cholesterol and lowering blood pressure.