CN110384709A - Composition and its application containing phloridzin and 1-DNJ - Google Patents

Abstract

The present invention relates to a kind of compositions of natural hypoglycemic active constituent, the combination includes the first component and the second component, wherein, it is selected from for first group: phloridzin, the plant extracts containing phloridzin or the material of vegetable origin containing phloridzin or the vegetable material by processing；And second group is selected from: 1-DNJ, the plant extracts containing 1-DNJ or the material of vegetable origin containing 1-DNJ or the vegetable material by processing.The composition can effectively reduce the taking dose of every kind of ingredient, reduce cost, while reducing and taking the issuable side effect of every kind of ingredient.

Description

Composition containing phlorizin and 1-deoxynojirimycin and application thereof

technical field

The invention relates to the fields of medicine, health products and food, in particular to a composition containing phlorizin and 1-deoxynojirimycin, and the application of the composition in lowering blood sugar.

Background technique

Diabetes Mellitus (DM) is a series of clinical syndromes caused by absolute or relative insufficiency of insulin in the body. The disease can also cause some complications during the development process, such as: hypoglycemia (Hypoglycemia), ketoacidosis (Ketoacidosis), nonketotic hyperosmolar coma (Nonketotic Hyperosmolar Coma), cardiovascular disease, chronic renal failure, retinal disease, neuropathy, and microvascular disease.

Commonly used drugs for diabetes treatment mainly include: biological drugs, such as insulin (Insulin); chemical drugs, such as sulfonylureas (Sulfonylurea), biguanides (Biguanides), and glitazones (Glitazone); and natural drugs, such as Chinese herbal medicines. Due to the lack of mature non-injection drug delivery technology, insulin is still mainly used by injection, and there are safety risks in the injection process. Some other injectable drugs, such as glucagon-like peptide 1 (GLP-1), fatty acylated glucagon-like peptide (NN2211, Novo Nordisk), Exendin-4 (Amylin) and poly Ethylene glycol-modified Exendin-4, etc., can effectively control blood sugar, reduce glycosylated hemoglobin HbA1c value and improve β-islet cell function after long-term administration, but because these molecules also act on central nervous receptors at the same time, they will produce Symptoms of vomiting and dizziness. Although some chemical drugs can lower blood sugar, they have different side effects or liver and kidney toxicity, and can easily lead to aggravation of cardiovascular complications. For example, DPP-IV inhibitor drugs, because they have an inhibitory effect on a large class of enzymes in the organism, will change the distribution of body fat, and the actual effect of administration is relatively poor. Although various Chinese patent medicines for treating diabetes have a certain therapeutic effect, it is difficult to guarantee the curative effect and quality of the medicine because the mechanism of action of the medicine is single or its mechanism of action and active ingredients are not clear.

In summary, there is an urgent clinical need to develop safer and more effective hypoglycemic drugs.

Contents of the invention

The object of the present invention is to provide a safe and effective formula of natural hypoglycemic active ingredient composition.

The first aspect of the present invention provides a composition of natural hypoglycemic active ingredients, said composition comprising a first component and a second component, wherein:

The first component is selected from the group consisting of: phlorizin, plant extracts containing phlorizin, or plant raw materials or processed plant materials containing phlorizin;

The second component is selected from: 1-deoxynojirimycin, plant extracts containing 1-deoxynojirimycin, or plant raw materials or processed plant materials containing 1-deoxynojirimycin.

In another preferred example, in the composition of hypoglycemic active ingredients, based on the weight of the two components of phlorizin and 1-deoxynojirimycin, the compatibility relationship is phlorizin: 1-deoxynojirimycin =(3～1000):1.

In another preferred example, in the composition of hypoglycemic active ingredients, based on the weight of the two components of phlorizin and 1-deoxynojirimycin, the compatibility relationship is phlorizin: 1-deoxynojirimycin =(10～500):1.

In another preferred example, in the composition of hypoglycemic active ingredients, based on the weight of the two components of phlorizin and 1-deoxynojirimycin, the compatibility relationship is phlorizin: 1-deoxynojirimycin =(100～500):1.

In another preferred example, in the composition of hypoglycemic active ingredients, based on the weight of the two components of phlorizin and 1-deoxynojirimycin, the compatibility relationship is phlorizin: 1-deoxynojirimycin =(100～300):1.

In another preferred example, in the composition of hypoglycemic active ingredients, the total weight of phlorizin and 1-deoxynojirimycin is 50-1000 mg/g.

In another preferred example, in the composition of hypoglycemic active ingredients, the total weight of phlorizin and 1-deoxynojirimycin is 100-980 mg/g.

In another preferred example, in the composition of hypoglycemic active ingredients, the total weight of phlorizin and 1-deoxynojirimycin is 300-980 mg/g.

In another preferred example, the composition of the hypoglycemic active ingredient is composed of phylloside and mulberry leaves containing phlorizin and mulberry leaves containing 1-deoxynojirimycin, the dry weight ratio of For: 1-100: 1-100.

In another preferred example, the hypoglycemic active ingredient composition is composed of Hubei crabapple leaves containing phlorizin and mulberry leaves containing 1-deoxynojirimycin, and the ratio of the dry weight of Hubei crabapple leaves to mulberry leaves is For: 1-100: 1-100.

In another preferred example, the dosage of the composition is such that the total mass of phlorizin and 1-deoxynojirimycin in the composition is ≤400 mg/kg body weight (preferably, ≤103 mg/kg body weight).

In another preferred example, the dosage of the composition is such that the mass of phlorizin in the composition is ≤300 mg/kg body weight (preferably, ≤100 mg/kg body weight).

In another preferred example, the dosage of the composition is that the mass of 1-deoxynojirimycin in the composition is ≤100 mg/kg body weight (preferably, ≤10 mg/kg body weight; more preferably, ≤3 mg/kg body weight ).

The second aspect of the present invention provides the use of the hypoglycemic active ingredient composition as described in the first aspect of the present invention, the composition is used for the preparation of animals and has (i) hypoglycemic effect, and/or (ii) Functional foods, health products and medicines for preventing and/or treating diabetes and its complications.

In another preferred example, the composition is used for the preparation of food, health products and other products that have the function of (i) lowering blood sugar, and/or (ii) preventing and/or treating diabetes and its complications. / or medication.

In the third aspect of the present invention, there is provided a method for (i) lowering blood sugar, and/or (ii) preventing and/or treating diabetes and its complications, including the step of: administering the first aspect to the object in need The composition of the natural hypoglycemic active ingredient.

In another preferred embodiment, the subject is an animal; preferably, a mammal; more preferably, a human.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, we will not repeat them here.

Description of drawings

Figure 1 is a diagram of the blood glucose levels of each group at each time point after a single oral administration of phlorizin to C57BL/6J mice.

Fig. 2 is a diagram of the area under the blood glucose curve of each group at each time point after a single oral administration of phlorizin to C57BL/6J mice.

Fig. 3 is a diagram of the blood glucose levels of each group at each time point after a single oral administration of 1-deoxynojirimycin C57BL/6J mice with starch.

Fig. 4 is a diagram of the area under the blood glucose curve of each group at each time point after a single oral administration of 1-deoxynojirimycin C57BL/6J mice with starch.

Fig. 5 is a diagram of the blood glucose levels of C57BL/6J mice administered starch at each time point by a single oral administration of phlorizin and 1-deoxynojirimycin prescription.

Figure 6 is a diagram of the area under the blood glucose curve of each group at each time point of starch administration to C57BL/6J mice after a single oral administration of phlorizin and 1-deoxynojirimycin.

Fig. 7 is a diagram of the blood glucose levels of each group at each time point after a single oral administration of phlorizin to C57BL/6J mice after starch was administered.

Figure 8 is a diagram of the area under the blood glucose curve of each group at each time point after a single oral administration of phlorizin to C57BL/6J mice after starch administration.

Fig. 9 is a diagram of the blood glucose levels of each group at each time point after a single oral administration of 1-deoxynojirimycin to C57BL/6J mice with starch.

Figure 10 is a diagram of the area under the blood glucose curve of each group at each time point after a single oral administration of 1-deoxynojirimycin to C57BL/6J mice after starch was administered.

Fig. 11 is a diagram of the blood glucose levels of each group at each time point after a single oral administration of phlorizin and 1-deoxynojirimycin prescription to C57BL/6J mice after starch administration.

Figure 12 is a diagram of the area under the blood glucose curve of each group at each time point after a single oral administration of phloridizin and 1-deoxynojirimycin prescription to C57BL/6J mice after starch administration.

Fig. 13 is a diagram of the blood glucose levels of each group at each time point after a single oral administration of sweet tea extract to C57BL/6J mice with starch.

Figure 14 is a diagram of the area under the blood glucose curve of each group at each time point after a single oral administration of sweet tea extract to C57BL/6J mice after starch administration.

Fig. 15 is a diagram of the blood glucose levels of each group at each time point after a single oral administration of mulberry leaf extract to C57BL/6J mice after starch was administered.

Figure 16 is a diagram of the area under the blood glucose curve of each group at each time point after a single oral administration of mulberry leaf extract to C57BL/6J mice after starch was administered.

Fig. 17 is a diagram of the blood glucose levels of each group at each time point after a single oral administration of sweet tea and mulberry leaf extract prescription to C57BL/6J mice after starch administration.

Figure 18 is a diagram of the area under the blood glucose curve of each group at each time point after a single oral administration of sweet tea and mulberry leaf extract prescription to C57BL/6J mice after starch administration.

Detailed ways

After long-term and in-depth research, the inventor unexpectedly discovered a safe and effective natural hypoglycemic active ingredient composition formulation, which contains phlorizin and 1-deoxynojirimycin. Compared with using phloridzin or 1-deoxynojirimycin alone, when the same hypoglycemic effect is achieved, the dosage of the composition of the present invention is significantly reduced; while taking phloridzin and 1-deoxynojirimycin containing no significant hypoglycemic effect dosage Nojirimycin can produce significant hypoglycemic effect, on this basis, the inventors have completed the present invention.

the term

Phlorizin

Phloridzin, English name: Phloridzin; molecular formula: C ₂₁ H ₂₄ O ₁₀ ; molecular weight: 436.41g/moL, easily soluble in hot water and ethanol, slightly soluble in cold water, insoluble in ether, chloroform, the structural formula of phloridzin As shown in formula (a). Phlorizin is the glucoside of phloretin, a dihydrochalcone substance belonging to flavonoids, and is a natural dual sodium-dependent glucose transporter (SGLT-1 and SGLT-2) inhibitor that reduces intestinal glucose Into the blood and inhibit the reabsorption of glucose by the renal tubules, so that the glucose with high concentration in the blood is excreted in the urine, thereby reducing blood sugar. Phlorizin can be extracted from natural plants or medicinal materials containing phlorizin (for example, plants such as Aegina officinale, apple tree, and Hubei crabapple).

1-deoxynojirimycin

1-deoxynojirimycin, English name: 1-deoxynojirimycin; molecular formula: C ₆ H ₁₃ NO ₄ ; molecular weight: 163.17; CAS number: 19130-96-2; soluble in water, dimethyl sulfoxide, methanol. 1-Deoxynojirimycin is found in mulberry, commelina, hyacinth and Adenophora. 1-Deoxynojirimycin is a natural alpha-glucosidase inhibitor that can delay the breakdown of starch in food into glucose in the intestine. Its structural formula is shown in formula b.

As used herein, "about", "approximately", and "approximately" refer to a value of ±10% (preferably, ±5%; more preferably, ±1%) centered on a certain point value. scope.

As used herein, the term "extract" or "plant extract" refers to an extract containing active ingredients (such as 1-deoxynojirimycin or phlorizin) obtained from plants as raw materials through conventional extraction methods such as solvent extraction, etc. substance. In this application, the extracts or plant extracts can be purchased from commercial channels, or prepared according to conventional methods, or prepared by referring to the preparation methods provided in this application.

As used herein, the terms "extract containing phlorizin" and "plant extract containing phlorizin" are used interchangeably and refer to plants derived from plants containing phlorizin (e.g. ) or a certain part of the plant (for example, Leucophyllum foliage, Keye leaves, Hubei Begonia leaves) are used as raw materials, and the substances containing phlorizin are extracted by conventional methods. Preferably, it is alcohol extract. Preferably, in the extract containing phlorizin, the content (mass fraction) of phlorizin is ≥ 10% (preferably, 20-99%; more preferably, 20-50%; most Preferably, 30±6% (30±3%)).

As used herein, the terms "1-deoxynojirimycin-containing extract" and "1-deoxynojirimycin-containing plant extract" are used interchangeably and refer to plants containing 1-deoxynojirimycin (e.g. , Mulberry, Commelina, Hyacinth and Adenophora) or a certain part of the plant as raw materials to obtain substances containing 1-deoxynojirimycin. Preferably, in the extract containing 1-deoxynojirimycin, the content (mass fraction) of 1-deoxynojirimycin ≥ 5%; preferably, about 5-80%; more preferably, about 5-30%; optimally, 10±3% (10±1%).

In the present application, the preparation method of the phlorizin-containing extract is not particularly limited. can use. The extract can be obtained by conventional methods, using Aegae officinalis, apple tree, Crabapple or parts thereof (for example, leaves of Aegina officinalis). In a preferred example, the extract containing phlorizin is obtained by solvent extraction. Preferably, the solvent is an alcoholic solvent, more preferably, ethanol.

As used herein, the term "plant raw material or processed plant material containing phlorizin" refers to the original plant or a part thereof containing phloridin, or the original plant or a part thereof containing phloridin parts of processed products. Preferably, the original plants containing phlorizin include: Aegae officinale, apple trees and/or Crabapples.

As used herein, the term "plant raw material or processed plant material containing 1-deoxynojirimycin" refers to the original plant or a part thereof containing 1-deoxynojirimycin, or the Processed products of the original plant or a part thereof. Preferably, the original plants containing 1-deoxynojirimycin include: mulberry, commelina, hyacinth and plants of the genus Adenophora.

As used herein, the term "prevention" refers to various means or measures used to prevent the occurrence or development of diseases before they are identified by clinical standards, including medical, physical or chemical methods, to prevent and reduce various diseases Onset or progression of symptoms.

As used herein, the term "diabetes prevention" refers to the use of the composition of the present invention for potential patients who have not met the clinical indicators of "diabetes" and will slowly develop into clinically defined "diabetes" as time goes on, thus Improve these patients' tolerance to glucose, promote the body's ability to metabolize glucose, and increase the body's sensitivity to insulin. Such potential patients usually suffer from "Metabolic Syndrome", such as: obesity, insulin resistance, glucose intolerance, hypertension, antherosclerosis, dyslipidemia (dyslipidemia) (i.e. high triglyceride levels, low high-density lipoprotein at the same time) and so on.

As used herein, the term "treatment" means to prevent and reduce the occurrence or development of the disease, so that the development or aggravation of the disease course can be inhibited, curbed, alleviated, improved, slowed down, stopped, delayed or reversed, the described maintenance and Various indicators of a disease, disorder or pathological state when administered include alleviation or reduction of symptoms or complications, or cure or elimination of the disease, disorder or condition, and/or administration.

As used herein, the term "treating diabetes" refers to using the composition of the present invention for patients clinically diagnosed as "diabetes", improving the tolerance of these patients to glucose, promoting the body's ability to metabolize glucose, and increasing the body's sensitivity to insulin Therefore, the patient's postprandial and fasting blood sugar can be controlled at a normal level. As the ability to metabolize glucose is improved, the occurrence and development of various cardiovascular diseases, chronic renal failure, retinopathy, neuropathy and microangiopathy caused by long-term high blood sugar are slowed down.

As used herein, the term "foodstuff" refers to a single compound or composition made edible including various compounds, compositions or extracts provided by the present invention. The production and manufacture of the single compound or composition should comply with relevant food safety standards, but these food safety standards should not limit the present invention.

As used herein, the term "health product" refers to an edible single compound or composition made of various compounds, compositions or extracts provided by the present invention to be administered to patients to prevent and treat diseases. Purpose. It belongs to the term food as used herein, but its production, manufacture and sale should also comply with various relevant requirements, standards and norms.

As used herein, the term "drug" refers to a single compound, a composition of multiple compounds, plants or Chinese medicinal materials and their extracts that can be used to prevent or treat a certain disease, or refers to a drug with a single compound as the main active ingredient. Composition or formulation (formulation) also refers to a composition or formulation with multiple compounds as active ingredients. "Drug" should be understood not only as a product that is approved and approved for production by the administrative agency established by it according to the laws and regulations of a country, but also refers to a product containing a single compound as the active ingredient formed in the process of obtaining approval and approval for production. various material forms. "Formation" should be understood as obtaining through chemical synthesis, biological transformation or purchase.

Composition of natural hypoglycemic active ingredients

The present invention provides a composition of natural hypoglycemic active ingredients, comprising a first component and a second component, wherein:

The first component is selected from: phlorizin, plant extracts containing phlorizin, or plant raw materials or processed plant materials containing phloridizin;

The second component is selected from: 1-deoxynojirimycin, plant extracts containing 1-deoxynojirimycin, or plant raw materials or processed plant materials containing 1-deoxynojirimycin.

In another preferred example, in the composition, based on the weight of the two components of phlorizin and 1-deoxynojirimycin, the compatibility relationship is phlorizin: 1-deoxynojirimycin=(3～ 1000):1.

In another preferred example, in the composition, based on the weight of the two components of phlorizin and 1-deoxynojirimycin, the compatibility relationship is phlorizin: 1-deoxynojirimycin=(10～ 500): 1.

In another preferred example, in the composition, based on the weight of the two components of phlorizin and 1-deoxynojirimycin, the compatibility relationship is phlorizin: 1-deoxynojirimycin=(100～ 500): 1.

In another preferred example, in the composition, based on the weight of the two components of phlorizin and 1-deoxynojirimycin, the compatibility relationship is phlorizin: 1-deoxynojirimycin=(100～ 300):1.

A composition of natural hypoglycemic active ingredients, wherein the total weight of phlorizin and 1-deoxynojirimycin is 50-1000 mg/g; - The total weight of deoxynojirimycin is 50-1000 mg.

In another preferred example, the total weight of phlorizin and 1-deoxynojirimycin is 100-980 mg/g.

In another preferred example, the total weight of phlorizin and 1-deoxynojirimycin is 300-980 mg/g.

A composition of natural hypoglycemic active ingredients, wherein, the composition is composed of phlox leaves containing phloridizin and mulberry leaves containing 1-deoxynojirimycin, and the dry weight of leaves and mulberry leaves is The ratio is: 1-100:1-100.

In another preferred example, the composition is composed of Hubei crabapple leaves containing phlorizin and mulberry leaves containing 1-deoxynojirimycin, and the dry weight ratio of Hubei crabapple leaves to mulberry leaves is 1-100 :1-100.

In another preferred example, the dosage (single dosage) of the composition is that the total mass of phlorizin and 1-deoxynojirimycin in the composition is ≤400 mg/kg body weight (preferably, ≤103 mg/kg body weight) .

In another preferred example, the dosage (single dosage) of the composition is such that the mass of phlorizin in the composition is ≤300 mg/kg body weight (preferably, ≤100 mg/kg body weight).

In another preferred example, the dosage (single dosage) of the composition is that the mass of 1-deoxynojirimycin in the composition is ≤100 mg/kg body weight (preferably, ≤10 mg/kg body weight; more preferably, ≤3mg/kg body weight).

In another preferred embodiment, the composition is an extract composition, that is, the first component of the composition is a plant extract containing phlorizin, and the second component is a plant extract containing 1-deoxynojirimycin of plant extracts. Preferably, in the extract composition, the phlorizin content of the first component is 30±6wt% and the 1-deoxynojirimycin content of the second component is 10±3wt%. Preferably, in the extract composition, the mass ratio of the first component to the second component is (10-100):1.

In another preferred example, the dosage (single dosage) of the extract composition is about 303-330 mg/kg body weight, based on the total mass of the first component and the second component. Preferably, the dosage of the extract composition is 300 mg/kg body weight of the first component and about 3-30 mg/kg body weight of the second component.

The present invention also provides a pharmaceutical composition comprising: (i) a safe and effective amount of the natural hypoglycemic active ingredient as described in the first aspect, and (ii) pharmaceutically acceptable pharmaceutical excipients. The pharmaceutical excipients can be conventionally used in various preparations (such as, but not limited to, isotonic agents, buffers, flavoring agents, excipients, fillers, binders, disintegrants and lubricants, etc. ); it can also be selected to be compatible with the substance (for example, but not limited to emulsifiers, solubilizers, bacteriostats, analgesics and antioxidants, etc.); this type of adjuvant can effectively improve the active composition The stability and solubility of the contained compound or change the release rate and absorption rate of the compound, etc., thereby improving the metabolism of the compound in the active composition in the living body, thereby enhancing the administration effect of the composition. In addition, the pharmaceutical adjuvant can also be an adjuvant (such as, but not limited to, gelatin, Albumin, chitosan, polyether and polyester polymer materials such as polyethylene glycol, polyurethane, polycarbonate and its copolymers, etc.). The main manifestations that are beneficial to drug administration include, but are not limited to: improving therapeutic effect, increasing bioavailability, reducing toxic and side effects, and improving patient compliance.

The pharmaceutical composition of the present invention can also be prepared into dosage forms that are beneficial to administration by conventional methods; said dosage forms include (but are not limited to): water pills, powders, tablets, granules, capsules, slow-release agents, controlled-release agent etc. The excipients used to prepare oral liquid preparations generally include: solvents, and optional flavoring agents, bacteriostats, emulsifiers, and coloring agents. The excipients used to make tablets generally include fillers (such as starch, powdered sugar, dextrin, lactose, compressible starch, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate and mannitol, etc.), binders (eg, ethanol, starch slurry, sodium carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, gelatin solution, sucrose solution, and aqueous solutions of polyvinylpyrrolidone or alcohol solution, etc.), disintegrants (such as dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone and cross-linked sodium carboxymethyl cellulose) and lubricants (such as: Magnesium stearate, micronized silica gel, talc, hydrogenated vegetable oil, macrogol 4,000, macrogol 6,000, magnesium lauryl sulfate, etc.), etc. The excipients used to make granules are similar to tablets, but the granulation process is different. According to needs, the prepared granules are mixed with a glidant, and then filled into capsules to obtain capsules.

use

A composition of natural hypoglycemic active ingredients is used for the preparation of food, health products and medicines with the functions of (i) lowering blood sugar, and/or (ii) preventing and/or treating diabetes and its complications.

In another preferred example, the use of the hypoglycemic active ingredient composition is to prepare food, health products and drugs.

The present invention also provides a method for (i) lowering blood sugar, and/or (ii) preventing and/or treating diabetes and its complications, comprising the steps of:

Administering a safe and effective amount of the composition of natural hypoglycemic active ingredients as described in the first aspect to a subject in need.

Wherein, "safe and effective amount" refers to: the amount of active ingredients (ie, phlorizin and 1-deoxynojirimycin) is sufficient to significantly improve the condition of diabetes and/or lower blood sugar without causing serious side effects.

In another preferred example, the administration (administration) method includes: oral administration.

Main advantage of the present invention:

(1) The composition can effectively reduce the blood sugar concentration, and can significantly reduce the dosage compared with taking phlorizin and 1-deoxynojirimycin alone.

(2) The composition delays the digestion of food to produce glucose and reduces the absorption of glucose; when the blood sugar concentration is too high, the glucose in the blood is excreted through the urine, more effectively maintaining the blood sugar at a normal level.

(3) Taking a small amount of the composition can achieve the effect of taking more phloridizin or 1-deoxynojirimycin alone, which can reduce costs and reduce drug side effects.

In the examples of this application, the mulberry leaf extract containing 1-deoxynojirimycin used was purchased from Shanghai Zishi Biotechnology Co., Ltd. (the content of 1-deoxynojirimycin was 10.1%); the 1-deoxynojirimycin used Pure Nojirimycin was purchased from Chengdu Pusi Biotechnology Co., Ltd. (content >99%).

Embodiment 1: Preparation of phlorizin and plant extract containing phlorizin

1 kg of Ginger Leaf Ke Ye (also known as sweet tea) crushed into coarse powder is extracted with 10 liters of 80% ethanol under reflux for 1 hour, the extract is filtered, and the residue is extracted with 10 liters of 80% ethanol under reflux for 1 hour, and the two extracts are combined , concentrated under reduced pressure to 1 liter, filtered the insoluble matter in the remaining aqueous liquid, and the gained filtrate was spray-dried to obtain 150 grams of extract containing phloridzin (detected by HPLC-UV, wherein the content of phloridzin was 29.4%). Get 50 grams of the above-mentioned extract containing phlorizin and dissolve it with 500 milliliters of 30% ethanol, put it on the HZ-801 type macroporous resin column (50 × 8cm column bed), and dissolve it with 30%, 60%, 90% ethanol-water respectively 5 liters of gradient elution, each collection of 500 ml, the eluate was detected by silica gel thin layer (developing solvent: chloroform-methanol-water 3:1:0.1), combined in 60% ethanol-water elution fraction containing root The fraction of dermoside was concentrated under reduced pressure to obtain an amorphous powder. Dissolve with a small amount of hot water, let cool to precipitate crystals, and obtain 10 grams of pure phloridin (detected by HPLC-UV, the content of phloridin is >98%).

Embodiment 2: Preparation of phlorizin (Phl) and 1-deoxynojirimycin (DNJ) monomer composition

Weigh phlorizin and 1-deoxynojirimycin crystals respectively, mix according to the weight ratio of 100:0.3, 100:1, 100:3, 20:1, 10:1, 5:1 and 3:1, add a small amount Dissolved in water for oral administration.

Example 3: Preparation of a plant extract containing phlorizin and a composition containing 1-deoxynojirimycin plant extract

Weigh the plant extract containing phlorizin and the plant extract containing 1-deoxynojirimycin respectively, mix them according to the weight ratio of 300:3, 300:10 and 300:30, add a small amount of water to dissolve, and use for gastric administration medication. Wherein, in the plant extract containing phlorizin, the mass fraction of phlorizin is 29.4%; in the plant extract containing 1-deoxynojirimycin, the mass fraction of 1-deoxynojirimycin is 10.1%.

Embodiment 4: Phlorizin (Phl) pharmacodynamics experiment in vivo

Effects of Phlorizin on Oral Glucose Tolerance in Normal C57BL/6J Mice

Animals: 21 C57BL/6J mice (8-10 weeks old, sex: male, body weight 18-22 g, purchased from Shanghai Lingchang Biological Co., Ltd.). Test substance dose and animal grouping: phlorizin 100 mg/kg (n=7); phlorizin 300 mg/kg (n=7); solvent control ultrapure water (n=7).

Procedure: The mice were starved for 12 hours, had free access to water, and were randomly divided into 3 groups according to body weight and starvation blood sugar. The mice were given intragastric administration, 15 minutes after the administration, 3 g/kg of glucose was administered by intragastric administration, the blood glucose was measured before, 15, 30, 60, 90 minutes and 2 hours after the administration of the sugar, and the administration time, body weight and blood glucose were recorded. Table 1 shows the effect (mmol/L) of oral administration of phlorizin on the oral glucose tolerance of normal C57BL/6J mice.

Table 1

Note: *, p<0.05 compared with Vehicle; #, p<0.01, compared with Vehicle.

From Table 1 and Fig. 1 and Fig. 2, it can be known that the test substance phlorizin can dose-dependently reduce the blood glucose of mice after intragastric administration of sugar in 15 min and 30 min after the single administration of phlorizin, and the effective dose is 300 mg/ kg, the area under the curve also showed dose-dependent changes at each time point.

Embodiment 5: In vivo pharmacodynamics experiment of 1-deoxynojirimycin (abbreviated as DNJ)

Effect of 1-deoxynojirimycin (DNJ) on oral glucose tolerance in normal C57BL/6J mice

Animals: 40 C57BL/6J mice (8-10 weeks old, sex: male, body weight 18-22 g, purchased from Shanghai Lingchang Biological Co., Ltd.). Test substance dose and animal grouping: 1-deoxynojirimycin 15mg/kg (n=8); 1-deoxynojirimycin 30mg/kg (n=8); 1-deoxynojirimycin 60mg/kg (n=8); 8); 1-deoxynojirimycin 100 mg/kg (n=8); solvent control ultrapure water (n=8).

Procedure: The mice were starved for 12 hours, had free access to water, and were randomly divided into 5 groups according to body weight and starvation blood sugar. Mice were administered by intragastric administration, 15 minutes after administration, 3g/kg of starch was administered by intragastric administration, blood glucose was measured before, 15, 30, 60, 90 minutes and 2 hours after starch administration, and the administration time, body weight and blood glucose were recorded. -The effect (mmol/L) of deoxynojirimycin (DNJ) single gavage administration on the oral glucose tolerance of normal C57BL/6J mice is shown in Table 2.

Table 2

Note: *, p<0.05 compared with Vehicle; #, p<0.01, compared with Vehicle.

From Table 2 and Fig. 3 and Fig. 4, it can be known that the test substance 1-deoxynojirimycin is administered with starch 15 minutes after a single administration, and can dose-dependently reduce the blood glucose of mice after oral administration of starch at 15 minutes and 30 minutes. The lower area also showed dose-dependent changes at each time point, and the effective dose was less than or equal to 15mg/kg.

Example 6: In vivo pharmacodynamic experiment of phlorizin and 1-deoxynojirimycin prescription (abbreviated as Phl+DNJ)

Effects of Phlorizin and 1-Deoxynojirimycin Recipe (referred to as Phl+DNJ) on Oral Glucose Tolerance in Normal C57BL/6J Mice

Animals: 40 C57BL/6J mice (8-10 weeks old, sex: male, body weight 18-22 g, purchased from Shanghai Lingchang Biological Co., Ltd.). Test substance dose and animal grouping: phlorizin 300mg/kg+1-deoxynojirimycin 15mg/kg (Phl+DNJ) (n=8); phlorizin 300mg/kg+1-deoxynojirimycin 30mg/kg kg(Phl+DNJ)(n=8); Phlorizin 300mg/kg+1-deoxynojirimycin 60mg/kg(Phl+DNJ)(n=8); Phlorizin 300mg/kg+1-deoxynojirimycin Mycin 100 mg/kg (Phl+DNJ) (n=8); solvent control ultrapure water (n=8).

Procedure: The mice were starved for 12 hours, had free access to water, and were randomly divided into 5 groups according to body weight and starvation blood sugar. Mice were administered by intragastric administration, 15 minutes after administration, 3g/kg of starch was administered by intragastric administration, blood glucose was measured before, 15, 30, 60, 90 minutes and 2 hours after starch administration, and the administration time, body weight and blood glucose were recorded. -The effect (mmol/L) of deoxynojirimycin prescription on the oral glucose tolerance of normal C57BL/6J mice by single gavage administration is shown in Table 3.

table 3

Note: *, p<0.05 compared with Vehicle; #, p<0.01, compared with Vehicle.

It can be seen from Table 3 and Figure 5 and Figure 6 that a single dose of the test substance Phl+DNJ can dose-dependently reduce the blood glucose of mice after intragastric administration of starch for 15 minutes and 30 minutes, and the area under the curve also shows a dose-dependent increase at each time point. Changes (see Table 3, Figure 5, Figure 6). It shows that the test substance Phl+DNJ in the range of 300~400mg/kg total dose can significantly reduce the blood sugar of mice level, with a significant hypoglycemic effect.

Embodiment 7: In vivo pharmacodynamics experiment of phlorizin

Effects of phlorizin on oral glucose tolerance in normal C57BL/6J mice

Animals: 16 C57BL/6J mice (8-10 weeks old, sex: male, body weight 18-22 g, purchased from Shanghai Lingchang Biological Co., Ltd.). Test substance dose and animal grouping: phlorizin 100 mg/kg (n=8); solvent control group ultrapure water (n=8).

Procedure: The mice were starved for 12 hours, had free access to water, and were randomly divided into two groups according to body weight and starvation blood sugar. Mice were administered by intragastric administration, 15 minutes after administration, 3g/kg of starch was administered by intragastric administration, blood glucose was measured before, 15, 30, 60, 90 minutes and 2 hours after starch administration, and the administration time, body weight and blood glucose were recorded. Table 4 shows the effect (mmol/L) of dermatosin (Phl) on the oral glucose tolerance of normal C57BL/6J mice after a single intragastric administration.

Table 4

Note: *, p<0.05 compared with Vehicle; #, p<0.01, compared with Vehicle.

From Table 4 and Fig. 7 and Fig. 8, it can be known that the test substance phloridizin was administered with starch 15 minutes after a single administration, and the test substance phloridizin 100mg/kg dose group had no effect on the oral glucose tolerance of normal mice. No significant change (see Table 4, Figure 7, Figure 8). Based on the research results of Example 4, it is suggested that starch or glucose was administered orally to mice, and the test substance phlorizin at 100 mg/kg dose did not show a significant hypoglycemic effect.

Example 8: In vivo pharmacodynamics experiment of 1-deoxynojirimycin (abbreviated as DNJ)

Effect of 1-deoxynojirimycin (DNJ) on oral glucose tolerance in normal C57BL/6J mice

Animals: 40 C57BL/6J mice (8-10 weeks old, sex: male, body weight 18-22 g, purchased from Shanghai Lingchang Biological Co., Ltd.). Test substance dose and animal grouping: 1-deoxynojirimycin 4 groups 0.3, 1, 3, 10 mg/kg (n=8); solvent control group ultrapure water (n=8).

Procedure: The mice were starved for 12 hours, had free access to water, and were randomly divided into 5 groups according to body weight and starvation blood sugar. Mice were administered by intragastric administration, 15 minutes after administration, 3g/kg of starch was administered by intragastric administration, blood glucose was measured before, 15, 30, 60, 90 minutes and 2 hours after starch administration, and the administration time, body weight and blood glucose were recorded. -The effect (mmol/L) of deoxynojirimycin single gavage administration on the oral glucose tolerance of normal C57BL/6J mice is shown in Table 5.

table 5

Note: *, p<0.05 compared with Vehicle; #, p<0.01, compared with Vehicle.

From Table 5 and Fig. 9 and Fig. 10, it can be seen that the test substance 1-deoxynojirimycin is given starch by intragastric administration 15 minutes after a single administration, and the blood glucose of mice after intragastric administration of starch can be dose-dependently reduced for 15 minutes and 30 minutes, The area under the curve also showed dose-dependent changes at each time point. The effective dose was 10 mg/kg, and the hypoglycemic rate was 25.94% (AUC 0-2h). The lower doses of 0.3mg/kg, 1mg/kg and 3mg/kg had a slight decrease in blood glucose levels at 15 or 30 minutes after sugar administration, but the overall area under the curve did not show a significant effect.

Example 9: In vivo pharmacodynamic experiment of phlorizin and 1-deoxynojirimycin prescription (referred to as Phl+DNJ)

Effects of Phlorizin and 1-Deoxynojirimycin Recipe (referred to as Phl+DNJ) on Oral Glucose Tolerance in Normal C57BL/6J Mice

Animals: 32 C57BL/6J mice (8-10 weeks old, sex: male, body weight 18-22 g, purchased from Shanghai Lingchang Biological Co., Ltd.). Dosage of test substance and animal grouping: phlorizin 100mg/kg+1-deoxynojirimycin 0.3mg/kg (Phl+DNJ) (n=8); phlorizin 100mg/kg+1-deoxynojirimycin 1mg /kg (Phl+DNJ) (n=8); Phlorizin 100mg/kg+1-deoxynojirimycin 3mg/kg (Phl+DNJ) (n=8); Solvent control group ultrapure water (n=8 ).

Procedure: The mice were starved for 12 hours, had free access to water, and were randomly divided into 4 groups according to body weight and starvation blood sugar. Mice were administered by intragastric administration, 15 minutes after administration, 3g/kg of starch was administered by intragastric administration, blood glucose was measured before, 15, 30, 60, 90 minutes and 2 hours after starch administration, and the administration time, body weight and blood glucose were recorded. Table 6 shows the effect (mmol/L) of oral administration of dermatosin and 1-deoxynojirimycin on the oral glucose tolerance of normal C57BL/6J mice by single gavage administration.

Table 6

Note: *, p<0.05 compared with Vehicle; #, p<0.01, compared with Vehicle.

From Table 6 and Figure 11 and Figure 12, it can be seen that the test substance Phl+DNJ can dose-dependently reduce the blood glucose of mice after intragastric administration of starch for 15 minutes and 30 minutes, and the area under the curve also shows dose-dependent changes at each time point. The effective dose is Phl+DNJ (100+0.3) mg/kg, and the hypoglycemic rate is 17.71% (AUC 0-2h) (see Table 6, Figure 11, Figure 12). Comprehensive embodiment 7 and the single drug single administration glucose tolerance test result of embodiment 8, Phl+DNJ (100+0.3) mg/kg combined administration reflects and is significantly better than phlorizin 100mg/kg and DNJ 0.3mg/kg. kg's hypoglycemic effect, suggesting that the combination of drugs is the superimposed effect of the two single drugs, or even a synergistic effect. The above results show that the administration of Phl+DNJ prescription keeps the total dose range of 100mg/kg within the range of 300:1～30:1, which is the curative effect range.

Example 10: In vivo pharmacodynamics experiment of sweet tea extract (29.4% phlorizin content)

Oral glucose tolerance test of sweet tea extract in normal C57BL/6J mice

Animals: 24 C57BL/6J mice (8-10 weeks old, sex: male, body weight 18-22 g, purchased from Shanghai Lingchang Biological Co., Ltd.). Test substance dose and animal grouping: sweet tea extract 100, 300 mg/kg (n=8); solvent control ultrapure water (n=8).

Procedure: The mice were starved for 12 hours, had free access to water, and were randomly divided into 3 groups according to body weight and starvation blood sugar. The mice were given intragastric administration, 15 minutes after the administration, 3 g/kg of starch was administered by intragastric administration, the blood glucose was measured before, 15, 30, 60, 90 minutes and 2 hours after the administration of the starch, and the administration time, body weight and blood glucose were recorded. Table 7 shows the effect (mmol/L) of oral administration of sweet tea extract on oral glucose tolerance of normal C57BL/6J mice.

Table 7

Note: *, p<0.05 compared with Vehicle; #, p<0.01, compared with Vehicle.

From Table 7 and Fig. 13 and Fig. 14, it can be seen that: the sweet tea extract of the test substance was intragastrically administered starch 15 minutes after a single administration, and compared with the normal control group, the 300mg/kg dosage group decreased by 60 and 120min after intragastric administration of starch. The area under the curve also decreased significantly (P=0.02) when the effective dose was 300mg/kg (see Table 7, Figure 13, Figure 14). Because the content of phloridzin contained in sweet tea extract is 29.4%, the content of phloridzin in 300 mg/kg sweet tea extract is about 100 mg/kg. The onset dose of sweet tea extract is 300mg/kg, and the effect is slightly better than the hypoglycemic effect of a single administration of phloridzin 100mg/kg (Example 7), suggesting that other ingredients contained in the extract may play a superimposed or synergistic role. Hypoglycemic effect of dermoside.

Example 11: In vivo pharmacodynamics experiment of mulberry leaf extract (1-deoxynojirimycin content 10%)

Effect of Mulberry Leaf Extract on Oral Glucose Tolerance in Normal C57BL/6J Mice

Animals: 16 C57BL/6J mice (8-10 weeks old, sex: male, body weight 18-22 g, purchased from Shanghai Lingchang Biological Co., Ltd.). Test substance dose and animal grouping: mulberry leaf extract 3 mg/kg (n=8); solvent control group ultrapure water (n=8).

Procedure: The mice were starved for 12 hours, had free access to water, and were randomly divided into two groups according to body weight and starvation blood sugar. The mice were given intragastric administration, 15 minutes after the administration, 3 g/kg of starch was administered by intragastric administration, the blood glucose was measured before, 15, 30, 60, 90 minutes and 2 hours after the administration of the starch, and the administration time, body weight and blood glucose were recorded. Table 8 shows the effect (mmol/L) of oral glucose tolerance of normal C57 mice by single gavage administration of mulberry leaf extract.

Table 8

Note: *, p<0.05 compared with Vehicle; #, p<0.01, compared with Vehicle.

From Table 8 and Fig. 15 and Fig. 16, it can be seen that the test substance mulberry leaf extract has no significant effect on the oral glucose tolerance of normal C57BL/6J mice after a single administration of the mulberry leaf extract for 15 minutes (see Table 8, Fig. 15, Figure 16). It is consistent with the result that a single administration of 0.3 mg/kg single component of DNJ has no hypoglycemic effect.

Example 12: In vivo pharmacodynamics experiment of sweet tea extract (phlorizin content 29.4%) and mulberry leaf extract (1-deoxynojirimycin content 10%)

Oral glucose tolerance test of sweet tea extract and mulberry leaf extract in normal C57BL/6J mice

Animals: 32 C57BL/6J mice (8-10 weeks old, sex: male, body weight 18-22 g, purchased from Shanghai Lingchang Biological Co., Ltd.). Test substance dose and animal grouping: sweet tea extract+mulberry leaf extract (abbreviation: sweet+mulberry) 300+3, 300+10, 300+30mg/kg (n=8); solvent control group ultrapure water (n =8).

Procedure: The mice were starved for 12 hours, had free access to water, and were randomly divided into 4 groups according to body weight and starvation blood sugar. The mice were given intragastric administration, 15 minutes after the administration, 3 g/kg of starch was administered by intragastric administration, the blood glucose was measured before, 15, 30, 60, 90 minutes and 2 hours after the administration of the starch, and the administration time, body weight and blood glucose were recorded. Table 9 shows the effect (mmol/L) of sweet tea extract and mulberry leaf extract (referred to as sweet + mulberry) group by intragastric administration on oral glucose tolerance of normal C57BL/6J mice.

Table 9

Note: *, p<0.05 compared with Vehicle; #, p<0.01, compared with Vehicle.

From Table 9 and Fig. 17 and Fig. 18, it can be seen that each dosage group of sweet tea extract+mulberry leaf extract (sweet+mulberry) can dose-dependently reduce mouse blood glucose and the area under the blood glucose curve at each point; The effective dose of sweet + mulberry is (300+3) mg/kg (see Table 9, Figure 17, Figure 18). The hypoglycemic effect of the prescription sweet + mulberry with an effective dose of (300+3) mg/kg is significantly better than that of sweet tea extract 300 mg/kg and mulberry leaf extract 3 mg/kg, suggesting that the prescription of mulberry leaf extract is beneficial to sweet tea Hypoglycemic efficacy of extracts. When the total dose of the existing sweet tea extract and mulberry leaf extract is within the range of 300-303 mg/kg, and the formula ratio is maintained at 100:1-10:1, or even lower, the hypoglycemic effect can be achieved.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (10)

1. a kind of composition of natural hypoglycemic active constituent, which is characterized in that the composition includes the first component and second Component, in which:

First group is selected from: phloridzin, the plant extracts containing phloridzin or material of vegetable origin or process containing phloridzin The vegetable material of processing；

Second group is selected from: 1-DNJ, the plant extracts containing 1-DNJ are wild containing 1- deoxidation The material of vegetable origin of buttocks mycin or the vegetable material by processing.

2. a kind of blood-sugar decreasing active composition as described in claim 1, which is characterized in that wild with phloridzin and 1- deoxidation The weight of two kinds of ingredients of buttocks mycin calculates, and compatibility relationship is phloridzin: 1-DNJ=(3~1000): 1.

3. a kind of blood-sugar decreasing active composition as claimed in claim 2, which is characterized in that wild with phloridzin and 1- deoxidation The weight of two kinds of ingredients of buttocks mycin calculates, and compatibility relationship is phloridzin: 1-DNJ=(10~500): 1.

4. a kind of blood-sugar decreasing active composition as claimed in claim 2, which is characterized in that wild with phloridzin and 1- deoxidation The weight of two kinds of ingredients of buttocks mycin calculates, and compatibility relationship is phloridzin: 1-DNJ=(100~300): 1.

5. a kind of blood-sugar decreasing active composition as described in claim 1, which is characterized in that phloridzin and 1- deoxidation open country buttocks Mycin adds up to weight to be 50~1000mg/g.

6. a kind of blood-sugar decreasing active composition as claimed in claim 5, which is characterized in that phloridzin and 1- deoxidation open country buttocks Mycin adds up to weight to be 100~980mg/g.

7. a kind of blood-sugar decreasing active composition as claimed in claim 5, which is characterized in that phloridzin and 1- deoxidation open country buttocks Mycin adds up to weight to be 300~980mg/g.

8. a kind of composition as described in claim 1, which is characterized in that by the lithocarpus litseifolius leaf containing phloridzin and contain 1- The mulberry leaf of 1-Deoxynojirimycin form, the ratio between lithocarpus litseifolius leaf and the dry weight of mulberry leaf are as follows: 1-100:1-100.

9. a kind of composition as described in claim 1, which is characterized in that by the Hubei Chinese flowering crabapple leaf containing phloridzin and contain 1- The mulberry leaf of 1-Deoxynojirimycin form, the ratio between Hubei Chinese flowering crabapple leaf and the dry weight of mulberry leaf are as follows: 1-100:1-100.

10. a kind of purposes of the blood-sugar decreasing active composition as described in claim 1~9, which is characterized in that be used to prepare Applied to animal there is (i) to reduce blood glucose, and/or (ii) prevents and/or treat the food of the function of diabetes and its complication Product, health care product and/or drug.