WO2018090983A9

WO2018090983A9 - Saponin compound for improving intestinal microflora, preparation method and use thereof

Info

Publication number: WO2018090983A9
Application number: PCT/CN2017/111690
Authority: WO
Inventors: 刘洁
Original assignee: 北京工商大学
Priority date: 2016-11-17
Filing date: 2017-11-17
Publication date: 2018-08-09
Also published as: CN107753547A; CN107753547B; WO2018090983A1

Abstract

Disclosed are a saponin compound for improving intestinal microflora, and a preparation method and use thereof, wherein the saponin compound is gypenoside. The composition can improve the intestinal microflora and promote colonization by characteristic microorganisms.

Description

Saponin compound for improving intestinal flora, preparation method and application thereof

Technical field

The invention belongs to the technical field of saponin compounds and preparation and application thereof. Specifically, the present invention relates to a triterpenoid saponin compound extracted and isolated from Gynostemma pentaphyllum, and a preparation method and use thereof.

Background technique

The human intestine is a complex, active, and relatively balanced system that colonizes a large number of complex microbial communities. The human gastrointestinal tract is home to about 10 ⁴ normal bacteria, of which 30-40 dominant bacteria constitute 99% of the total intestinal bacteria. Intestinal microbes contain a wide variety of enzyme systems that can participate in a range of physiological processes such as host energy, material and genetic information transfer. Many studies have shown that there is an inseparable mutual symbiotic relationship between the intestinal flora and the host. The intestinal microbes can form a biological barrier in the intestinal tract, through mass occupying, nutrient competition and various metabolites secreted by them. And bacteriocins play an important role in the digestion, absorption, immune response and metabolic activity of the host. At the level of the door, Firmicutes and Bacteroides are two dominant species; at the genus level, Bacteroides, Clostridium, Clostridium, and Bifidobacteria are four types closely related to human health. Related dominant bacteria are also the focus of research on intestinal micro-organisms at this stage. The occurrence and development of chronic diseases such as obesity and hyperlipidemia are closely related to the structural imbalance of intestinal flora.

Gynostemma pentahyllum (Thunb.) Makino is a perennial herbaceous genus of the genus Cucurbitaceae, and its main bioactive component is a triterpenoid saponin.

In view of the high medicinal value of triterpenoid saponins, further research is needed in the art.

Summary of the invention

It is an object of the present invention to provide a saponin compound, a preparation method and application thereof.

In a first aspect of the invention, there is provided a use of a Gynostemma total saponin compound,

(1) a composition for preparing an improved intestinal flora; and/or

(2) For the preparation of a composition that promotes the colonization of characteristic microorganisms.

In another preferred embodiment, the composition is a pharmaceutical composition or a food composition.

In another preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

In another preferred embodiment, the pharmaceutical composition is an oral agent.

In another preferred embodiment, the dosage form of the pharmaceutical composition is selected from the group consisting of a tablet, a capsule or a granule.

In another preferred embodiment, the intestinal flora belongs to the phylum of the genus Bacteroides or the bacillus.

In another preferred embodiment, the intestinal flora belongs to the sputum microphylaxis or the proteobacteria.

In another preferred embodiment, the intestinal flora is selected from the group consisting of lactic acid bacteria, bifidobacteria, or Clostridium natto.

In another preferred embodiment, the intestinal flora is Akkermansia muciniphila.

In another preferred embodiment, the food composition is a diet food.

In another preferred embodiment, the pharmaceutically acceptable salt of the total saponin compound is selected from the group consisting of sodium salts, potassium salts, or combinations thereof.

In another preferred embodiment, the Gynostemma pentaphyllum is a diploid Gynostemma pentaphyllum or a tetraploid Gynostemma pentaphyllum, preferably a tetraploid Gynostemma pentaphyllum.

In another preferred embodiment, the total saponin compound (designated YJ) comprises one or more compounds selected from the group consisting of:

According to a second aspect of the present invention, there is provided a method for non-therapeutic improvement of intestinal flora in vitro comprising the step of administering a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt thereof, at a locus to be treated.

In another preferred embodiment, the compound or a pharmaceutically acceptable salt thereof is administered at a concentration of 0.001 to 0.02 μg/ml, preferably 0.01 to 0.15 μg/ml, more preferably 0.05. -0.1 μg/ml.

In a third aspect of the invention, a probiotic composition is provided, the probiotic composition comprising:

(1) a first drug: a saponin compound, and

(2) a second drug: one or more probiotics selected from the group consisting of lactic acid bacteria, bifidobacteria, Clostridium genus, Akkermansia muciniphila, or a combination thereof;

Wherein the first drug and the second drug are located in the same or different containers.

In another preferred embodiment, the mass ratio of the first drug to the second drug (in terms of active ingredient) in the probiotic composition is 1:100-100:1, preferably 1: 10-10:1, more preferably 1:5-5:1.

In another preferred embodiment, the probiotic composition is a pharmaceutical composition or a food composition.

According to a fourth aspect of the invention, there is provided a method of improving intestinal flora comprising the step of administering a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt thereof, to a subject in need thereof.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

DRAWINGS

Figure 1 shows the preventive effects of different doses of total saponin compound YJ on obesity caused by a high-fat diet.

A stands for low-fat diet; B stands for high-fat diet; C stands for high-fat group + low-dose YJ (100 mg/Kg); D stands for high-fat group + high-dose YJ (300 mg/Kg), 8 mice per group, * indicates that the low-fat group has a significant body weight compared with the high-fat group, and # indicates that the total saponin compound intervention group is extremely significant at the P<0.05 level compared with the high-fat group.

Figure 2 shows the effect of total saponin compound YJ on community diversity of intestinal bacteria at the gate level.

A represents low-fat diet feeding; B represents high-fat diet feeding; C stands for high-fat group + low-dose YJ (100 mg/Kg), and D stands for high-fat group + high-dose YJ (300 mg/Kg).

Figure 3 shows the effect of total saponin compound YJ on community diversity of gut bacteria at the genus level, A for low fat feed; B for high fat diet; C for high fat + low dose YJ (100 mg/Kg) D represents a high fat group + a high dose YJ (300 mg/Kg).

Figure 4 shows the effect of PCOA on total intestinal saponin compound YJ on intestinal flora-genus, A for low-fat diet feeding; B for high-fat diet; C for high-fat group + low-dose YJ (100 mg/kg), D Represents high fat group + high dose YJ (300mg/Kg).

Figure 5 shows the effect of total saponin compound YJ on the intestinal flora-gate. HF stands for high fat diet obesity model group; HF+YJ stands for high fat diet +300 mg/kg dose YJ.

Figure 6 shows the effect of total saponin compound YJ on the diversity of intestinal bacteria in obese mice at the gate level, HF for the high fat diet obesity model group (mouse number HF 1-8); HF+YJ stands for high Fat feed + 300 mg/kg dose YJ (mouse number HF_YJ 1-8).

Figure 7 shows the effect of total saponin compound YJ on the microbial diversity of intestinal bacteria in obese mice, HF represents the high fat diet obesity model group (mouse number HF 1-8); HF+YJ stands high Fat feed + 300 mg/kg dose YJ (mouse number HF_YJ 1-8).

Figure 8 shows the effect of PCA analysis of total saponin compound YJ on intestinal flora diversity in obese mice, HF for high fat diet obesity model group (mouse number HF 1-8); HF+YJ for high fat diet +300 mg/kg dose YJ (mouse number HF_YJ 1-8).

detailed description

The inventors of the present application have extensively and intensively studied the chemical constituents and biological activities of Gynostemma pentaphyllum, and successfully isolated a plurality of new triterpenoid saponins from Gynostemma pentaphyllum, and first discovered the triterpene total saponin compound YJ and One or more monomeric compounds have a function of preventing obesity by improving the intestinal flora of the animal. On the basis of this, the present invention has been completed.

Terminology

All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, unless otherwise defined.

As used herein, when used in reference to a particular recited value, the term "about" means that the value can vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values between 99 and 101 and (eg, 99.1, 99.2, 99.3, 99.4, etc.).

As used herein, the terms "containing" or "including" may be open, semi-closed, and closed. In other words, the terms also include "consisting essentially of," or "consisting of."

Gynostemma

Gynostemma pentaphyllum is a dry whole grass of Gynostemma pentaphyllum (Thunb. Makino), which is a perennial climbing herb. Gynostemma pentaphyllum has a long history of eating and was first used in the Ming Dynasty's "Rescue Materia Medica" for wild vegetables. The Japanese folks call it "Gancha Man", a sweet tea substitute or a sweetener for diabetics. Gynostemma pentaphyllum is also one of the plants with ginseng saponin resources outside Wujiake. It has many functions such as nourishing health, anti-cancer, anti-aging, blood fat reduction and blood sugar lowering. It is known as “Southern Ginseng”. Gynostemma pentaphyllum is one of the plants that can be used for health care products announced by the Ministry of Health. Because of its remarkable health care effect and high safety, it is suitable for long-term use. It is often used as a lipid-lowering health tea in the folk.

Total saponin compound YJ

The total saponin compound YJ of the present invention includes, but is not limited to, a saponin compound extracted from Gynostemma pentaphyllum and the like, that is, a series of compounds having a saponin structure.

In another preferred embodiment, the total saponin compound of the present invention comprises one or more compounds selected from the group consisting of:

Intestinal bacteria and obesity

The composition of intestinal flora is closely related to obesity. Studies at the animal level indicate that compared with normal mice, ob/ob obese mice have increased the number of bacteria in the intestinal flora, and the number of Bacteroides Reduced by 50%; at the human level, 12 fat patients were followed up for treatment with a fat-restricted diet and a carbohydrate-restricted diet. After one year, the weight of the former decreased by 2%, and the weight of the latter decreased by 6%. The 16s rRNA gene study showed that the total amount of energy in the food did not change, and the number of thick-walled bacteria in the intestine after losing weight through two diets. Decreased, while the number of Bacteroides gates increased. The data suggest that an increase in the proportion of Bacteroides/Thick Wall bacteria in the gut contributes to the prevention and treatment of obesity.

Akkermansia muciniphila

Akkermansia muciniphila belongs to the family Verrucomicrobiaceae. In May 2013, Belgian researchers published an article on PNAS to discover a "slimming bacteria" by studying a gut bacteria. The liquid medium containing this bacteria can be greatly Change the health of obese mice. Researchers at the Catholic University of Leuven, Belgium, fed a bacterium called Akkermansia muciniphila two to three times more fat than normal mice, and their excess weight was reduced by nearly half without changing their diet. And the main symptoms of type 2 diabetes in the body, insulin antibody levels are also reduced. Studies have shown that Akkermansia muciniphila increases intestinal levels of endogenous cannabinoids that control glucose and intestinal balance and impedes diet-induced reduction in mucosal barrier thickness. In the same year, researchers published an article on Gut, and metformin significantly improved Akkermansia. The number of Muciniphila is used to improve the blood glucose profile of high fat fed mice. Compared with the high-fat-fed group, metformin-treated high-fat-fed mice had more mucin-degrading bacteria Akkermansia. In addition, the number of goblet cells producing mucin in the metformin-treated group was also significantly increased. Oral administration of Akkermansia Muciniphila to high-fat-fed mice significantly improved glucose tolerance even without taking metformin, which attenuated adipose tissue inflammation by inducing Foxp3 regulatory T cells (Tregs) in visceral adipose tissue. Therefore, the use of such bacteria to prevent or treat obesity and type 2 diabetes has become a potential strategy for the prevention and treatment of obesity.

Preparation

In another preferred embodiment, the method for preparing a triterpenoid saponin compound of the present invention comprises the following steps:

(1) extracting the aerial part of the tetraploid Gynostemma pentaphyllum with a solvent, and concentrating the extract to obtain an extract;

(2) Dissolving the extract with water to obtain a dispersion, and sequentially extracting with petroleum ether, ethyl acetate and n-butanol, and extracting to obtain an n-butanol extract, and recovering the solvent under reduced pressure to obtain an n-butanol extract;

(3) After dissolving the n-butanol extract water, apply it to the D101 macroporous resin column, first elute with pure water to remove sugar and water-soluble impurities, and then elute with ethanol-water gradient to collect different elution sites. , tracking detection, rotary evaporation concentrated volume, to obtain crude saponin dry powder. Then, the dissolved pure methanol solution was applied to a Sephadex LH-20 separation column, and the gradient elution was carried out. The solvent was recovered under reduced pressure, and then subjected to Sephadex LH-20 Sephadex gel chromatography, and eluted isocratically with a methanol solution. The fraction of the compound is separated and purified directly by semi-preparative reversed-phase high performance liquid chromatography to obtain the compound;

In another preferred embodiment, in the step (1), the aerial part of the tetraploid Gynostemma pentaphyllum is extracted with a solvent for 2-5 times, each time 1-3 hours, and the extract is concentrated to obtain an extract;

In another preferred embodiment, the solvent in the step (1) is selected from the mixture of one or more of methanol, ethanol, acetone or water;

In another preferred embodiment, the mass ratio of the extract to the water in the step (2) is 1:2-1:15.

In another preferred embodiment, the ratio of the amount of petroleum ether added to the dispersion in the step (2) is 1:2-5:1;

In another preferred embodiment, the ratio of the amount of ethyl acetate added to the dispersion in the step (2) is 1:2-5:1;

In another preferred embodiment, the volume ratio of n-butanol to the dispersion in the step (2) is 1:2 to 5:1.

In another preferred embodiment, in the step (3), the ethanol-water solution is subjected to a gradient elution at a volume ratio of 20:1 to 100:1; and then separated by Sephadex LH-20 column chromatography, and eluted sequentially with a pure methanol solvent. Collecting fractions containing the compound; one fraction is separated and purified by semi-preparative reversed-phase high performance liquid chromatography in a solvent system of 30% acetonitrile-15% methanol-55% water (vol) to obtain saponin compound 1 in sequence -9.

use

By investigating the effects of the total saponin compounds of the present invention on the proportion of the intestinal flora of the genus Bacteroides and the thick-walled bacteria, especially the slimming bacteria (Akkermansia muciniphila) in the genus Microsporaceae can be significantly improved, and the mechanism of action thereof is studied. It was found that the compounds of the present invention and their optical isomers, pharmaceutically acceptable salts and solvates can be significantly adjusted high. The intestinal flora caused by the fat diet to prevent obesity.

In addition, the compounds of the present invention can also be used as food adjuvants or food additives, added to foods for improving the intestinal flora of animals and promoting the colonization of beneficial microorganisms.

Pharmaceutical composition

The invention also provides a pharmaceutical composition comprising an active ingredient in a safe and effective amount, together with a pharmaceutically acceptable carrier.

The "active ingredient" as used in the present invention means the Gynostemma total saponin compound of the present invention.

The "active ingredients" and pharmaceutical compositions of the present invention are useful for improving intestinal flora composition and promoting beneficial microbial colonization.

In another preferred embodiment, it is used to prepare a medicament which improves the intestinal flora of an animal.

By "safe and effective amount" is meant that the amount of active ingredient is sufficient to significantly improve the condition without causing serious side effects. In general, the pharmaceutical compositions contain from 1 to 2000 mg of active ingredient per dose, more preferably from 10 to 200 mg of active ingredient per dose. Preferably, the "one dose" is a tablet.

"Pharmaceutically acceptable carrier" means: one or more compatible solid or liquid fillers or gel materials which are suitable for human use and which must be of sufficient purity and of sufficiently low toxicity. By "compatibility" it is meant herein that the components of the composition are capable of intermingling with the active ingredients of the present invention and with respect to each other without significantly reducing the efficacy of the active ingredients.

Typically, the carrier includes one or more of a diluent, a filler, a disintegrant, a lubricant, a colorant, a flavoring agent, or other conventional additives.

Examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid). , magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier (such as

), a wetting agent (such as sodium lauryl sulfate), a coloring agent, a flavoring agent, a stabilizer, an antioxidant, a preservative, a pyrogen-free water, and the like.

The mode of administration of the active ingredient or pharmaceutical composition of the present invention includes oral administration and the like.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.

In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) a disintegrant such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent such as paraffin; (f) Absorbing accelerators, for example, quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, hard Calcium citrate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or a mixture thereof. In capsules, tablets and pills, the dosage form may also contain a buffer.

The solid dosage forms can also be prepared with coatings and shell materials, such as casings and other materials known in the art. They can An opacifying agent is included and the release of the active ingredient in such a composition can be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs. In addition to the active ingredient, the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances. In addition to these inert diluents, the compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.

In addition to the active ingredient, the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.

Compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

The compounds of the invention may be administered alone or in combination with other therapeutic agents.

When a pharmaceutical composition is used, a safe and effective amount of a compound of the invention is administered to a mammal (e.g., a human) in need of treatment wherein the dosage is a pharmaceutically effective effective dosage, for a 60 kg body weight, The dose to be administered is usually from 1 to 2000 mg, preferably from 20 to 500 mg. Of course, specific doses should also consider factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled physician.

The above-mentioned features mentioned in the present invention, or the features mentioned in the embodiments, may be arbitrarily combined. All of the features disclosed in the present specification can be used in combination with any of the compositions, and the various features disclosed in the specification can be replaced by any alternative feature that provides the same, equal or similar purpose. Therefore, unless otherwise stated, the disclosed features are only general examples of equal or similar features.

The main advantages of the invention are:

(1) The present invention obtains a novel triterpenoid saponin compound by extracting and separating from tetraploid Gynostemma pentaphyllum, and the compound and their optical isomers, pharmaceutically acceptable salts and solvates thereof have been used for improving intestinal flora of animals. To promote the use of beneficial microbial colonization and to prevent and treat obesity, which may be administered in its own form or in combination with a pharmaceutically acceptable carrier;

(2) The present invention provides a simple and easy preparation method, which can be completed in a general-purpose device;

(3) The prepared compound has high purity, good economic benefit and application prospect.

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the manufacturer. Percentage and number of parts are weight unless otherwise stated Percentage and parts by weight.

The experimental materials and reagents used in the following examples are available from commercially available sources unless otherwise specified.

Example 1

1.1 Preparation of tetraploid Gynostemma total saponins YJ

The 18kg tetraploid Gynostemma pentaphyllum leaves were weighed by a pulverizer and then extracted by 20L 95% ethanol reflux method (feed ratio: 1:2.6), extracted 3 times for 3h, 2h, 1h, and removed by rotary evaporation. Ethanol to obtain crude extract of Gynostemma pentaphyllum

paste.

The above extract was dispersed in water, and successively extracted with an equal volume of petroleum ether, ethyl acetate and n-butanol to obtain petroleum ether, ethyl acetate, n-butanol and water extracting sites, respectively. Rotary evaporation to obtain the n-butanol fraction of the crude extract of Gynostemma pentaphyllum

Cream 1509g.

Prepare 6mg/mL n-butanol sample solution for a total volume of 20mL, load D101 macroporous resin, elute with 20%, 40%, 60% ethanol gradient, collect 60% elution site, and concentrate by rotary evaporation to obtain Dry saponin dry powder. Then, 15 g of crude saponin was dissolved in 55 mL of methanol solution and Sephadex LH-20 separation column was further purified by gradient elution, and monitored in real time, and the sample was collected to obtain a refined saponin total saponin compound YJ.

1.2 Animal experiment

SPF grade 5 week old male C57/BL 6 mice (purchased from Beijing Vitallihua Experimental Animal Center) were raised in an SPF environment.

1.2.1 Prevention experiment

After 32 mice were acclimated for one week, they were divided into 4 groups, 8 in each group, and fed in a single cage.

They were group A (low-fat group, 10% energy derived from fat), group B (high-fat group, 45% energy derived from fat), group C (high-fat group +100 mg/Kg/d YJ), and group D (High fat group +300mg/Kg/d YJ).

The breeding environment temperature is 22±2°C, the humidity is 30-70%, the mice are free to eat and drink water; the experiment lasts for 12 weeks, the weight is recorded every day, and the food and sterile water are renewed every Tuesday and Friday. Record the amount and record the amount of food consumed. Change the mouse padding weekly and change the cage every month. After 12 weeks, 3 days of fecal volume was collected continuously. Four mice were randomly selected from each group, DNA was extracted, and Illumina Miseq PE250/PE300 was sequenced to analyze the bacterial diversity of the fecal samples.

1.2.2 animal treatment experiment

The mice were fed with high-fat diet for 4 months and weighed about 40 g. After obesity-resistant mice were removed, they were randomly divided into 2 groups, 8 in each group, and fed in single cages.

The model group (HFD) continued to feed high-fat diet. In the intervention group (HFD+YJ), in addition to feeding high-fat diet, the tube was given a total dose of 300 mg/Kg/d total saponin compound for 2 months, and the body weight was recorded every day. New food and sterile water will be renewed on Tuesdays and Fridays, and a weighing record will be made to record the food intake. Change the mouse padding weekly and change the cage every month. After 2 months, all mice were continuously collected for 3 days of fecal volume, and DNA was extracted for Illumina Miseq PE250/PE300. Sequencing, analysis of the diversity of fecal sample flora.

1.3 Analysis of results

As can be seen from Figure 1, the long-term consumption of high-fat diet compared with low-fat diet, the weight of the mice gradually increased, reaching a significant level after 56 days, after adding YJ intervention, compared with the high-fat model group, the body weight The increase was strongly inhibited, reaching a significant level after 63 days and exhibiting a concentration dose relationship.

It can be seen from Fig. 2 that compared with the low-fat diet, long-term consumption of high-fat diet can cause a significant decrease in the proportion of the intestinal flora of Bacteroides/Thick-wall bacteria. After the intervention of YJ, the Bacteroides gate/thick wall The proportion of the bacteria began to rise and reached a significant increase under high dose conditions. Moreover, the high-dose intervention group induced Verrucomicrobiaceae and increased the number of Proteobacteria. Further classification of gut bacteria-genus, high-dose total saponin compounds can significantly induce a significant increase in Akkermansia muciniphila, a slimming bacterium in the genus Microflora (Fig. 3). Statistical analysis of PCOA, total saponin compound intervention can significantly improve the intestinal composition changes induced by high-fat diet in mice (Figure 4).

As can be seen from Fig. 3, YJ has a great influence on the diversity of the following strains: Bacteroides, Akkermansia muciniphila, lactic acid bacteria and the like.

It can be seen from Fig. 5 that after adding 28 mg/kg YJ to obesity for 28 days, the body weight showed a significant decrease, which proved that YJ has a significant therapeutic effect on obesity.

The results of the animal treatment experiment are shown in Fig. 6. It can be seen that the YJ intervention mouse, Verrucomicrobiaceae (blue mark) was significantly induced to express, further at the genus level, further proved the total saponin compound weight loss And its induction to Akkermansia muciniphila. Therefore, YJ is a prebiotic substance that specifically promotes the proliferation of Akkermansia muciniphila. Administration of 300 mg/kg YJ significantly increased the content of Akkermansia muciniphila compared to the control group.

Figure 7 shows the effect of YJ on the diversity of intestinal bacteria in obese mice at the genus level. Similarly, Akkermansia muciniphila was significantly improved. By PCA analysis, the two groups could be completely separated, demonstrating that YJ is regulating obese mice. The composition of the intestinal flora constitutes a significant effect on the prevention and treatment of obesity (Fig. 8).

In combination with the aforementioned preventive experiments, we can judge that the effect of total saponin compounds on the prevention and treatment of obesity is due to the targeted induction of "lean bacteria" Akkermansia muciniphila.

Example 2: Food

Prepare diet foods according to the following formula:

2-5 parts by weight of oatmeal, 1-5 parts by weight of yam, 10-15 parts by weight of dietary fiber, 1-4 parts by weight of glutinous rice, 2-5 parts by weight of inositol, 10-15 parts by weight of maltose, YJ 0.1%-0.5 % by weight

3 meals a day for 7-15 consecutive days.

Example 3: Pharmaceutical composition

The above components are uniformly mixed and directly compressed, that is, a tablet composition is obtained, one tablet per oral administration, 2-3 times a day.

Example 4: Pharmaceutical composition

The above component mixture was uniformly filled into 1000 capsules, one tablet per oral administration, 2-3 times a day.

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the appended claims.

Claims

Use of a Gynostemma total saponin compound, characterized in that

(1) a composition for preparing an improved intestinal flora; and/or

(2) For the preparation of a composition that promotes the colonization of characteristic microorganisms.
The use according to claim 1, wherein the intestinal flora belongs to a thick-walled fungus or a bacillus.
The use according to claim 1, wherein the intestinal flora belongs to the sputum microphylaxis or the proteobacteria.
The use according to claim 1, wherein the intestinal flora is selected from the group consisting of lactic acid bacteria, bifidobacteria or Clostridium.
The use according to claim 1, wherein the intestinal flora is Akkermansia muciniphila.
The use according to claim 1, wherein the composition is a pharmaceutical composition or a food composition.
The use according to claim 1, wherein the Gynostemma pentaphyllum is diploid Gynostemma pentaphyllum or tetraploid Gynostemma pentaphyllum, preferably tetraploid Gynostemma pentaphyllum.
The use according to claim 1, wherein the total saponin compound comprises one or more compounds selected from the group consisting of:
A method of non-therapeutically improving intestinal flora in vitro comprising the step of administering a compound of claim 1 or a pharmaceutically acceptable salt thereof at a locus to be treated.
A probiotic composition comprising:

(1) a first drug: a saponin compound, and

(2) a second drug: one or more probiotics selected from the group consisting of lactic acid bacteria, bifidobacteria, Clostridium genus, Akkermansia muciniphila, or a combination thereof;

Wherein the first drug and the second drug are located in the same or different containers.