CN1965873A - Chinese medicinal extract having blood sugar-lowering activity, its preparation process, composition and use - Google Patents

Abstract

The invention discloses a hypoglycemic extract extracted from a plant of the genus Balaophora (Balaophora L), a preparation method of the extract, a pharmaceutical composition containing a plant of the genus Balaophora (Balaophora L) or the extract of the plant of the genus, and snake Use of Balaophora L or its extracts and pharmaceutical compositions as medicines and health products, especially for the preparation of medicines and health products for treating and preventing diabetes and its complications and other related metabolic diseases .

Description

A traditional Chinese medicine extract with hypoglycemic activity, preparation method, composition and application

technical field

The present invention relates to extracting an extract capable of lowering blood sugar from a plant of the genus Balaophora (Balaophora L), a preparation method of the extract, a pharmaceutical composition containing a plant of the genus Balaophora (Balaophora L) or an extract of the genus, and snake The use of the plant of the genus Balaophora (Balaophora L) or the extract of the genus and its pharmaceutical composition as medicine, especially for the preparation of medicines and health products for treating and preventing diabetes and its complications and other related metabolic diseases.

Background technique

According to the relevant data of WHO, the prevalence rate, disability rate, mortality rate and overall health hazard of diabetes have been ranked third among non-communicable diseases. It has become a serious threat to human health after cardiovascular and cerebrovascular diseases and tumors. the third leading disease. There are 130 million people with diabetes in the world today, more than 90% of them are not type 2 diabetes patients, and it is increasing at a rate of 1% every year, and it is expected to soar to 230 million by 2020. The total number of diabetic patients in my country has exceeded 60 million, and the annual incidence rate is expected to exceed 6%. The disease not only brings great pain to the patients, but also greatly affects the quality of life, even threatens the lives of the patients, and also brings a heavy economic burden to the society. Both clinicians and patients hope for more and more effective treatments. At present, oral drugs for clinical treatment of diabetes include sulfonylureas, biguanides, thiazolidinedione insulin sensitizers, non-sulfonylurea insulin secretory agents, and α-glucosidase inhibitors.

Sulfonylureas include toluenesulfonylurea, chlorpropamide, glibenclamide, glipizide, and gliclazide. In addition to the high failure rate of the first treatment, these drugs are prone to secondary failure, and about 10% of patients do not respond to subsequent treatment. The incidence of adverse reactions is relatively high, such as allergies, dizziness, liver damage, mainly hypoglycemic shock and weight gain, especially chlorpropamide and glibenclamide are more common, so overweight patients should not use these drugs.

Biguanides include metformin and phenformin. Although this type of drug has been used to treat diabetes in the late 1950s, the incidence of adverse reactions is very high, mainly including obvious digestive tract side effects such as gastrointestinal irritation and lactic acid poisoning, which limit its application. In particular, phenformin has been banned in many countries because of its serious adverse reactions.

Representative drugs that have been marketed as insulin sensitizers include thiazolidinediones such as rosiglitazone. Thiazolidinedione drugs can directly enhance the insulin sensitivity of liver, muscle and adipose tissue in patients with type 2 diabetes, and the main target is peroxisome proliferator-activated receptor gamma (PPARγ). Patients' fasting and postprandial blood glucose decreased without causing hypoglycemia. In recent years, it has developed rapidly, and troglitazone, rosiglitazone, and pioglitazone have been listed successively, but troglitazone has been withdrawn from the market due to severe hepatotoxicity (liver failure).

A short-acting oral insulin secretion enhancer includes repaglinide and nateglinide, which mainly act on the ATP-sensitive potassium channel of the beta cell membrane of the pancreas to promote the secretion of insulin by the beta cell. Taking it with meals can correct the abnormal insulin secretion pattern related to eating in patients with type 2 diabetes, simulating physiological insulin secretion. It has a quick onset of action and a short duration of action. It only has a strong stimulating effect on the secretion of insulin during meals, so that patients can achieve good overall blood sugar control. The incidence of severe hypoglycemia was significantly lower than that of sulfonylureas. It is excreted in feces and has little liver and kidney toxicity, but it is contraindicated for patients with β-cell failure, diabetic ketoacidosis and severe liver and kidney damage.

The α-glucosidase inhibitors have been marketed as acarbose, voglibose and miglitol. With the listing of Bayer Acatbose (Acatbose) in the early 1990s, it has been recognized as a new way to treat diabetes at home and abroad, and its main target is the small intestine, so Acatbose has become the first-line drug for treating diabetes. And further expand the indications.

Among the plants of the genus Balaophora (Balaophora L) in the family Balaophoraceae (Balaophoraceae), the medicinal raw materials are Pueraria japonica (Balaophora japonica Mak.), Balaophora inovolucrata Hook f. and Balaophora inovolucrata Hook f. (Balaophorapolyandra Griff.).

Kudzu mushrooms (Balaophora japonica Mak.), Balaophora japonica Hook f. and Balaophora polyandra Griff. are perennial parasitic succulent herbs. Rhizomes grow underground and are spherical in shape with small nodules on the surface. The flower stem is red or yellow, and there are fat spikes on it. The flowers are unisexual, monoecious or dioecious, and bloom in August. It is a dioecious (unisexual inflorescence) plant, and multiple massive rhizomes are gathered together. All without chlorophyll. Stem slender, leaves alternate or nearly opposite. The male inflorescence is elliptical, and there is a bract under the male flower. The bracts fuse with each other to form many hexagonal nests, and the stalked male flowers grow from the base of the nests, and the tepals are 3 bases. Female inflorescences are nearly spherical or elliptic, with small rods obconically shaped, apex truncated, 3-4 times as long as wide; female flowers are only attached to the main axis of the inflorescence. September-October results. The male flowers of Snakeweed have stalks, and the middle part of the flower stem is covered by a leaf-shaped sheath; while the male flowers of Snakeweed are sessile, and the middle of the flower stem has no sheath. It grows in the hillside forest at an altitude of 1000--2500 meters, and parasitizes on the roots of woody plants.

The medicine has the effects of hemostasis, muscle growth and analgesia. Folks are often used to treat stomach pain, epistaxis, women's menstrual bleeding, dysentery and traumatic bleeding. It can also be used as a tonic. It can be decocted to relieve drunkenness, powdered and mixed with sesame oil for external use to treat rectal prolapse.

The pharmacological effects of Balaophora L have been reported to have anti-inflammatory, analgesic and anti-alcoholic effects [1, Ruan Hanli, Journal of Traditional Chinese Medicine, 2003, 21(6): 910-911] [2, Rong Juquan, Chinese Journal of National Folk Medicine, 2003, 65 (total): 347-349].

The compounds obtained in the study of the chemical constituents of the genus Balaophora (Balaophora L) are: β-amyresin (Balaophora A), palmitoyl lupine enol ester (Balaophora L), β-amylacetate, Lupine acetate, β-amyresinone, lupine ketone and palmitic acid, (-)-lariciresinol, (-)-pinoresinol [(-)-pinoresinol], 1-O-caffeoyl-β- D-glucopyranose, 1-O-caffeoyl-(6-O-galloyl)-β-D-glucopyranose, 1-O-caffeoyl-(4-O-galloyl)-β- D-Glucopyranose, gallic acid, coniferol, coniferol-3-O-(6-O-palmitoyl)-β-D-glucopyranoside, coniferin, methyl coniferin[3 , Shen Xiaoling, Chinese Herbal Medicine, 1996, 27 (5): 259-260] [4, Xia Xinzhong, Chinese Herbal Medicine, 2001, 32 (1: 6-9)] [5, Liu Xikui, Yunnan Plant Research, 1998, 20 (3): 369-373].

Contents of the invention

The object of the present invention is to provide the extract that extracts from the genus Balaophora (Balaophora L).

Another object of the present invention is to provide a method for preparing such an extract.

Another object of the present invention is to provide a pharmaceutical composition, including Balaophora L, extracts of Balaophora L, and carriers commonly used in the field of pharmacy.

Another object of the present invention is to provide a plant of the genus Balaophora (Balaophora L) and/or a plant of the genus Balaophora (Balaophora L), or an extract containing plants of the genus Balaophora (Balaophora L) and/or a plant of the genus Balaophora (Balaophora L) ) extract composition in the preparation of medicines for the treatment and prevention of diabetes and its complications.

Another object of the present invention is to provide a plant of the genus Balaophora (Balaophora L) and/or an extract of a plant of the genus Balaophora (Balaophora L), or a composition containing a plant of the genus Balaophora (Balaophora L) and/or an extract. Application in health care products for the treatment and prevention of diabetes and its complications.

Another object of the present invention is to provide a plant of the genus Balaophora (Balaophora L) and/or a plant of the genus Balaophora (Balaophora L), or an extract containing plants of the genus Balaophora (Balaophora L) and/or a plant of the genus Balaophora (Balaophora L) ) application of the composition of the extract in the preparation of medicines for the treatment and prevention of diabetes-related metabolic diseases.

Another object of the present invention is to provide a plant of the genus Balaophora (Balaophora L) and/or a plant of the genus Balaophora (Balaophora L), or an extract containing plants of the genus Balaophora (Balaophora L) and/or a plant of the genus Balaophora (Balaophora L) ) application of the composition of the extract in the preparation of health care products for the treatment and prevention of diabetes-related metabolic diseases.

In order to accomplish the object of the present invention, the present invention takes following technical scheme:

The raw medicinal material of Balaophora L. is dried and properly crushed to increase the contact area between the medicinal material and the solvent and improve the efficiency.

Water, alcohols, or a mixture of water and alcohols are used as solvents for extracting raw medicinal materials. Preferred alcohols include methanol, ethanol, isopropanol, butanol, and the like. A mixture of water and alcohols, such as water containing 40%-80% (volume ratio) of alcohol compounds. The amount of solvent during extraction is 4-14 times of the original drug weight. Extraction can be static or dynamic, preferably under dynamic conditions. In order to increase the extraction efficiency, ultrasonic waves or the like can be used. The temperature of the extraction is in the range from room temperature (eg 20° C.) to the reflux temperature of the solvent, preferably at the reflux temperature. Extraction can be carried out continuously or intermittently, and can be repeated 1-4 times during intermittent extraction.

After the above steps are completed, the filtrates are combined, and the filtrates are cooled under normal pressure or under reduced pressure, heated and concentrated to a volume 1-5 times the weight of the medicinal materials in a dynamic state. The extraction solvent used is an alcohol compound including methanol, ethanol, isopropanol, butanol, etc., or a mixture thereof; preferably ethanol. Let stand to precipitate, filter or centrifuge to remove insoluble matter, wash insoluble matter with water, generally 1-3 times. The combined filtrates were further concentrated into a paste.

Gained cream is purified, according to the extraction method described in claim 8, it is characterized in that, purification is by, solvent extraction, gel filtration, polyamide, macroporous resin, ion resin or adsorption column chromatography. The effective part obtained.

Adsorption column chromatography includes silica gel, alumina, cellulose, polyamide. The amount of adsorbent used is 30-200 times, preferably 80-100 times, more preferably 90-100 times of the sample amount. The elution system can be sieved by thin-layer chromatography, and a solvent system with an Rf value of 0.2-0.3 of the separated components is selected. Appropriate bases, such as ammonia, diethylamine, pyridine, 2-picoline, collidine, and N-ethylmorpholine, are added to the eluent to prevent tailing and promote separation. Activated carbon can be mixed with an appropriate amount of diatomaceous earth as a diluent to increase the flow rate of the solution.

The extract can also be directly purified and concentrated by exchange column and membrane technology, and then prepared into extract or dry powder. The exchange column that can be used includes: macroporous resin, ion exchange resin, activated carbon, dextran gel, etc.; preferably macroporous resin and activated carbon.

The extract can be freeze-dried into dry powder, or the concentrated liquid can be directly spray-dried into dry powder for various preparations.

The present invention also relates to a pharmaceutical composition containing the extract of the present invention as an active ingredient and conventional pharmaceutical excipients or adjuvants. Usually the pharmaceutical composition of the present invention contains 0.1-95% by weight of the extract of the present invention.

The present invention also provides a pharmaceutical composition, which includes an effective dose of medicine, as an active ingredient

According to the method of the present invention, Balaophora L and/or Balaophora L extract and a pharmaceutically acceptable carrier.

The pharmaceutical composition of the extract of the present invention can be prepared according to methods known in the art. When used for this purpose, if necessary, the extract of the present invention can be combined with one or more solid or liquid pharmaceutical excipients and/or adjuvants to make a suitable administration form that can be used as human medicine or veterinary medicine or dosage form.

The extract of the present invention or the pharmaceutical composition containing it can be administered in the form of a unit dose, and the route of administration can be enteral or parenteral, such as oral, nasal cavity, oral mucosa, skin, peritoneal or rectal administration, etc., preferably oral administration medicine.

The route of administration of the extract of the present invention or the pharmaceutical composition containing it also includes injection administration. Injection includes intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection and the like.

The dosage forms for administration may be liquid dosage forms or solid dosage forms. For example, the liquid dosage forms can be true solutions, colloids, particulate dosage forms, emulsion dosage forms, and suspension dosage forms. Other dosage forms such as tablets, capsules, drop pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, freeze-dried powder injections, etc.

The extract of the present invention can be made into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

Various carriers known in the art can be widely used for tableting unit dosage forms. Examples of carriers are, for example, diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, silicic acid Aluminum, etc.; wetting agents and binders, such as water, glycerin, polyethylene glycol, ethanol, propanol, starch paste, dextrin, syrup, honey, glucose solution, acacia mucilage, gelatin paste, sodium carboxymethylcellulose , shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.; disintegrants, such as dry starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene sorbate Sugar alcohol fatty acid esters, sodium lauryl sulfonate, etc.; lubricants, such as talc, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol, etc. Tablets can also be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

For example, in order to form a dosage unit into a pellet, various carriers known in the art can be widely used. Examples of carriers are, for example, diluents and absorbents, such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, kaolin, talc, etc.; binders, such as acacia, tragacanth, Gelatin, ethanol, honey, glucose solution, mucilage arabic, gelatin paste, sodium carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.; disintegrants such as dry starch, alginate , agar powder, algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfonate, methyl cellulose, ethyl cellulose, etc.; disintegration inhibition Agents, such as sucrose, glyceryl tristearate, cocoa butter, hydrogenated oil, etc.; absorption enhancers, such as quaternary ammonium salts, stearates, boric acid, liquid paraffin, polyethylene glycol, etc. Tablets can also be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

For example, in order to form a dosage unit into a pellet, various carriers known in the art can be widely used. Examples of carriers are, for example, diluents and absorbents such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, kaolin, talc, etc.; binders such as acacia, tragacanth, Gelatin, ethanol, honey, liquid sugar, rice paste, etc. Disintegrants, such as agar powder, dry starch, alginate, sodium dodecylsulfonate, methylcellulose, etc. For example, in order to form a dosage unit into a capsule, the active ingredient extract of the present invention is mixed with the above-mentioned various carriers, and the mixture thus obtained is placed in a hard gelatin capsule or a soft capsule. The active ingredient extract of the present invention can also be made into microcapsules, suspended in an aqueous medium to form a suspension, and can also be packed into hard capsules or made into injections for application.

For example, the extract of the present invention is made into injection preparations, such as solutions, suspension solutions, emulsions, and freeze-dried powder injections. This preparation can be aqueous or non-aqueous, and can contain one and/or more A pharmaceutically acceptable carrier, diluent, binder, lubricant, preservative, surfactant or dispersant. For example, the diluent may be selected from water, ethanol, polyethylene glycol, 1,3-propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid ester, and the like. In addition, in order to prepare an isotonic injection injection, an appropriate amount of sodium chloride, glucose or glycerin can be added to the injection preparation, and in addition, conventional co-solvents, buffers, pH regulators, etc. can also be added. These excipients are commonly used in the art. In addition, colorants, preservatives, fragrances, correctives, sweeteners or other materials can also be added to the pharmaceutical preparations, if necessary.

In order to achieve the purpose of medication and enhance the therapeutic effect, the medicine or pharmaceutical composition of the present invention can be administered by any known administration method.

The dosage of the extract pharmaceutical composition of the present invention depends on many factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, body weight, personality and individual reaction of the patient or animal, the route of administration, and the number of administrations , therapeutic purposes, so the therapeutic dosage of the present invention can vary widely. Generally speaking, the dosages of the pharmaceutical ingredients in the present invention are well known to those skilled in the art. According to the actual amount of drug contained in the final preparation of the extract composition of the present invention, it can be adjusted appropriately to meet the requirement of its therapeutically effective dose, so as to complete the preventive or therapeutic purpose of the present invention. The suitable daily dosage range of the extract of the present invention is 0.001-100 g/Kg body weight, preferably 0.01-50 g/Kg body weight, most preferably 0.05-25 g/Kg body weight. The above-mentioned dosage may be administered in a single dosage form or divided into several, such as two, three or four dosages, subject to the clinical experience of the administering physician and the dosage regimen including the use of other therapeutic means. The total dosage required for each treatment may be divided into multiple doses or administered in a single dose. The extract or composition of the present invention can be taken alone, or used in combination with other therapeutic drugs or symptomatic drugs and adjusted in dosage.

Description of drawings

Figure 1. Inhibitory effect of snake wild extract on blood glucose rise after oral administration of starch load in normal mice

Figure 2. Inhibitory effect of snake wild extract on blood glucose rise in normal mice after oral administration of sucrose load

Figure 3. The effect of snake wild extract on blood sugar rise after glucose load in normal mice

Figure 4. The effect of snake wild extract on insulin tolerance in normal mice

Detailed ways

The following examples specifically demonstrate the application of the present invention. But this embodiment does not limit the application scope of the present invention.

extraction experiment

Embodiment 1. water extraction extract

Pueraria japonica (Snake mushroom, Balaophora japonica Mak.) whole herb 100g, heat reflux with 500g distilled water, extract three times (500ml water/time), extract 1 hour each time, extract 26.0g water extract after decompression concentration, dissolve In 50 ml of water, add ethanol until the alcohol content reaches 70% (volume ratio), precipitate, filter, and concentrate the filtrate to dryness to obtain 14.0 g of a yellow powdery solid.

The filtrate part is separated by macroporous resin column:

14.0 g of the filtrate was dissolved in 50 ml of water, added to a 100 g macroporous resin column (S-8), washed with distilled water until the effluent was colorless, then eluted with ethanol until the effluent was colorless, and the ethanol eluted part was concentrated to obtain 7.5g. The macroporous resin is washed with water until the effluent is alcohol-free (macroporous resin regeneration).

Embodiment 2. alcohol extraction extract

2500g whole herb of Balaophora inovolucrata Hook f., heat reflux with 95% EtOH, extract three times (10L 95% EtOH/time), slowly heat to boiling, extract for 1 hour each time, and concentrate the extract under reduced pressure 532.0 g of 95% EtOH extract was obtained, suspended in 1500 ml of water, filtered, and the filtrate was extracted three times with ethyl acetate (1500 ml/time), the extracts were combined, and the solvent was recovered under reduced pressure to obtain 210.0 g of a yellow powdery solid.

Embodiment 3. ethyl acetate extraction extract

Balaophora polyandra Griff. (Balaophora polyandra Griff.) whole herb 1000g, heated to reflux with 5000ml of petroleum ether, extracted three times, 1 hour each time, the extract was concentrated under reduced pressure to obtain 56.0g of petroleum ether extract; 5000ml of the ester was heated to reflux, extracted three times, each time for one hour, the extracts were combined, and the solvent was recovered under reduced pressure to obtain 173.0g of ethyl acetate extraction fraction.

Ethyl acetate extraction part is separated by macroporous resin:

173.0 g of the filtrate was dissolved in 1000 ml of water, filtered, and the filtrate was added to a 1000 g macroporous resin column (S-8), washed with distilled water until the effluent was colorless, then eluted with ethanol until the effluent was colorless, and ethanol was eluted Partial concentration gave 113.0 g. The macroporous resin is washed with water until the effluent is alcohol-free (macroporous resin regeneration).

Pharmacological activity experiment:

Experimental example 1. Inhibition of the increase in blood sugar after starch load in normal mice by the extract of snake wild rice

Normal ICR male mice were randomly divided into 5 groups (n=10), fasted overnight before the experiment, one group was administered with starch (3.0g/kg) as a control group, and one group was treated with starch and Acarbose (Baitangping, 10mg/kg) by intragastric administration as the positive control drug group, and the remaining groups were administered intragastrically with starch (3.0g/kg) and different extracts (SHG88, SHG89, SHG90) of 20g crude drug/kg body weight respectively, before intragastric administration And 30min, 60min, 120min after gavage, blood was collected, and blood glucose concentration was determined by glucose oxidase method. The results (Fig. 1, Table 1) show that the extract of wild snakehead at 20g crude drug/kg dosage can significantly inhibit the increase of blood glucose after starch loading in normal mice, and the area under the curve (AUC) of blood glucose decreases. Different parts of the snake wild extract have different strengths. This result is similar to the effect of the α-glucosidase inhibitor Acarbose.

Table 1. The effect of Snake Wilderness Extract on the AUC of starch tolerance in normal mice AUC(mg/dl.h) ConSHG88SHG89SHG90Acarbose 385.9±69.2311.4±34.4 ^* 359.8±52.0357.8±71.6280.6±28.8 ^**

^* , ^** , compared with the control group, p<0.05, 0.01.

Experimental example 2. Inhibitory effect of snake wild extract on blood sugar rise after sucrose load in normal mice

Normal ICR male mice were randomly divided into 4 groups (n=10), fasted overnight before the experiment, one group was given sucrose (4.0g/kg) orally as a positive control group, and the remaining groups were treated with sucrose and different extracts Drugs (SHG88, SHG89, SHG90) were gavaged with 20 g crude drug/kg body weight, and the blood glucose concentration was measured before and 30 min, 60 min, and 120 min after gavage. The results (Fig. 2, Table 2) showed that the extract of wild snakehead at 20g crude drug/kg dose significantly inhibited the increase of blood sugar in normal mice after sucrose loading, the peak blood sugar moved back, and the area under the blood sugar curve (AUC) decreased. Different parts of the snake wild extract have different strengths.

Table 2. Effects of Snake Wilderness Extract on AUC of Normal Mice Sucrose Tolerance AUC(mg/dl.h) ConSHG88SHG89SHG90 277.5±52.5169.8±27.5 ^*** 181.1±18.8 ^*** 229.0±30.9 ^*

^* , ^*** , compared with the control group, p<0.05, 0.001.

Experimental example 3. The effect of snake wild extract on blood sugar rise after glucose load in normal mice

Normal ICR male mice were randomly divided into 4 groups (n=10), fasted for 2 hours before the experiment, one group was administered with glucose (2.0g/kg) as a control group, and the remaining groups were treated with glucose and different snake mushrooms. Extracts (SHG88, SHG89, SHG90) each 20g crude drug/kg body weight were gavaged continuously for 7 days, blood was collected before gavage and 30min, 60min, 120min after gavage, and blood glucose concentration was determined by glucose oxidase method. Result (Fig. 3, table 3) illustrates, and the extract of wild snakehead extract under 20g crude drug/kg dosage, obviously suppresses the raising of blood sugar after the oral glucose load of normal mouse, and blood sugar peak value reduces, and blood sugar area under the curve (AUC) reduce. It shows that the extract of Snakeweed not only has the function of inhibiting the activity of α-glucosidase and delaying the absorption of glucose, but also has the function of promoting glucose metabolism in normal mice. Different parts of the snake wild extract have different strengths.

Table 3. Effects of Snake Wilderness Extract on AUC of Glucose Tolerance in Normal Mice AUC(mg/dl.h) ConSHG88 343.8±67.2250.9±44.6 ^* SHG89SHG90 291.0±54.2287.0±46.7

^* , compared with the control group, p<0.05.

Experimental example 4. The effect of snake wild extract on insulin tolerance in normal mice

Normal ICR male mice were randomly divided into 4 groups (n=10), one group was given water as a control group, and the rest of the groups were treated with different extracts (SHG88, SHG89, SHG90) of 20g crude drug/kg body weight respectively Gavage for 10 consecutive days. Fasting for 2 hours before the experiment, insulin 0.4U/kg was injected subcutaneously, blood was collected before, 30 minutes and 60 minutes after the injection, and the blood glucose concentration was measured by the glucose oxidase method. Result (Fig. 4, table 4) shows that, the extract of wild snakehead extract under 20g crude drug/kg dose, obviously enhances the reactivity of normal mouse subcutaneous injection insulin, compares with corresponding control group, the blood glucose value of each time point is obvious The area under the curve (AUC) of blood glucose decreased. It shows that the extract of snake wild rice can increase the sensitivity of normal mice to insulin. Different parts of the snake wild extract have different strengths.

Table 4. The effect of snake wild extract on the AUC of insulin tolerance in normal mice AUC(mg/dl.h) ConSHG88SHG89SHG90 81.6±9.864.1±12.4 ^** 60.5±19.0 ^* 76.5±12.9

^* , ^** , compared with the control group, p<0.05, 0.01.

Experimental example 5. Inhibition of protein tyrosine phosphatase 1b (PTP1b) by the extract of wild snakehead

Protein tyrosine phosphatase 1b (PTP1b) is one of the important regulators in the insulin signaling system. Many factors in the insulin signal transduction system are activated by the protein tyrosine phosphorylation of their molecules, and then dephosphorylated and inactivated by PTP1b, so as to maintain the balance of the insulin signal transduction system. Therefore, PTP1b is considered to be one of the new targets of insulin sensitizers. We obtained the PTP1b protein by gene recombination, and observed in vitro experiments that 100 μg/ml of Snakeweed extract could significantly inhibit the activity of PTP1b (Table 5). Different parts of the snake wild extract have different strengths. The results of Experimental Examples 4 and 5 all indicate that the extract of Snakeweed has insulin-sensitizing effect.

Table 5. Inhibitory effect of snakehead extract on protein tyrosine phosphatase 1b (PTP1b) (n=3) Snake Wild Extract OD 405nm Inhibition rate(%) ConSHG08SHG09SHG17SHG18SHG88SHG89SHG90 0.321±0.0110.028±0.0030.136±0.0350.249±0.0120.090±0.0070.107±0.0480.175±0.0300.314±0.009 091.257.622.671.966.845.62.4

Experimental example 6. The effect of the extract of snake wild rice on the blood sugar of normal mice

Male mice with normal ICR were randomly divided into groups (n=10). The control group was fed with water every day, and the rest of the groups were given different 5, 20, 20, and 20 g crude drug extracts (SHG21, SHG88, SHG89, and SHG90) respectively. /kg body weight orally for 7-10 consecutive days. Blood was collected at 8:30 am on the day of the experiment, and no-fasting (no-fasting) blood sugar (equivalent to postprandial blood sugar) was measured. Fasting was performed for 2.5 hours on the day of the experiment, and fasting blood glucose was measured 2 hours after administration before blood collection. Blood glucose concentrations were determined by the glucose oxidase method. As a result, SHG21, SHG88, SHG89, and SHG90 respectively reduced the mean value of the animal's non-fasting (no-fasting) blood glucose by 27, 23, 16, and 6%; 25, 19, 7, 0%. It shows that the extract of wild snakehead has a certain effect on reducing the fasting and postprandial blood sugar of normal mice under the dose of 5-20g crude drug/kg, and the extracts of wild snakehead in different parts have different action strengths.

Experimental example 7. The effect of snake wild extract on the blood sugar of alloxan hyperglycemia mice

ICR male mice were injected with alloxan 75 mg/kg through the tail vein, and blood was taken to measure blood sugar 72 hours later. Those with blood glucose >200 mg/dl were hyperglycemic mice, and animals were grouped according to their blood glucose levels (n=10). The control group was gavaged with water every day, and the treatment group was gavaged with 5 g of crude drug/kg body weight of Snakeweed extract SHG21 for 10 consecutive days. Blood was collected at 8:30 am on the day of the experiment, and no-fasting (no-fasting) blood glucose (equivalent to postprandial blood glucose) was measured. Fasting for 2.5 hours on the day of the experiment, and measuring fasting blood glucose before blood collection 2 hours after administration. Blood glucose concentrations were determined by the glucose oxidase method. The results show that, under the dose of 5g crude drug/kg, the extract SHG21 of Snakeweed can reduce the mean value of non-fasting (no-fasting) blood sugar of alloxan hyperglycemia mice by 42%; The average is 34% lower. It shows that the extract of wild snake has a certain effect on reducing blood sugar in mice with hyperglycemia caused by alloxan.

The above 7 pharmacological experiments showed that the extract of Balaophora L had a certain activity of inhibiting α-glucosidase, so that the peak value of blood sugar rise after animal polysaccharide and disaccharide loading was reduced and moved back, and the blood sugar curve was lower. Reduced area; has a certain insulin sensitization effect, which can increase the range of insulin-induced blood sugar reduction in animals, and reduce the area under the blood sugar curve; shows a certain inhibitory effect on PTP1B; can promote glucose metabolism in the body, and increase blood sugar in animals after glucose load The peak value is reduced, and the area under the blood glucose curve is reduced; in addition, it also has a certain effect of reducing fasting blood glucose and non-fasting blood glucose in normal and alloxan hyperglycemic mice. These effects are all beneficial to the improvement of diabetes glucose metabolism and lipid metabolism disorders, and especially can be used to prepare medicines and health care products for treating and preventing diabetes and its complications and other related metabolic diseases.

Claims (13)

1. the application of Balanophora plant (Balaophora L) in preparation treatment and prevent diabetes and complication medicine thereof.

2. Balanophora plant according to claim 1 (Balaophora L), it is characterized in that, described crude drug is a Herba balanophorae japonicae (Balaenoptera borealis Lesson, Balaophora japonica Mak.), Balanophora involicrata Hook.f. (Balaophora inovolucrata Hook f.) and Herba Balanophorae polyandrae (Balaophorapolyandra Griff.).

3. the application of Balanophora plant (Balaophora L) in the medicine of preparation treatment and prevent diabetes correlated metabolism diseases.

4. the extracting method of a Balanophora plant (Balaophora L) extract is characterized in that, crude drug Balanophora plant (Balaophora L) through super-dry, pulverizing, is used solvent extraction, and merge extractive liquid, concentrates, and carries out purification again.

5. extracting method according to claim 4 is characterized in that, described pulverizing is that the gained granule size has been 200 mesh sieves from herb.

6. extracting method according to claim 4 is characterized in that described solvent is selected from organic solvent, water, mixed organic solvents, the mixture of water and organic solvent.

7. extracting method according to claim 4 is characterized in that, purification be by, solvent extraction, gel filtration, polyamide, macroporous resin, ion exchange resin or adsorpting column chromatography.

8. extracting method according to claim 4 is characterized in that, the optimal pH of the eluant during purification is 5-10.

9. extracting method according to claim 7 is characterized in that the macroporous resin resin is a polystyrene type resin, and described adsorpting column chromatography comprises silica gel, aluminium oxide, cellulose, polyamide, activated carbon.

10. Balanophora plant (BalaophoraL) extract that method as claimed in claim 4 is extracted.

11. pharmaceutical composition, it is characterized in that, comprise medicine effective dose, Balanophora plant (BalaophoraL) extract and/or Balanophora plant (Balaophora L) and the pharmaceutically acceptable carrier that extract as the method as claimed in claim 4 of active component.

12. the application of Balanophora plant (Balaophora L) extract that method as claimed in claim 4 is extracted in preparation treatment and prevent diabetes and complication medicine thereof.

13. the application of Balanophora plant (Balaophora L) extract that method as claimed in claim 6 is extracted in the preparation euglycemic agent.

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