CN111202752A - Ackermanella compositions - Google Patents
Ackermanella compositions Download PDFInfo
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- CN111202752A CN111202752A CN202010096535.9A CN202010096535A CN111202752A CN 111202752 A CN111202752 A CN 111202752A CN 202010096535 A CN202010096535 A CN 202010096535A CN 111202752 A CN111202752 A CN 111202752A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/13—Amines
- A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
- A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
Abstract
An Achroman bacterium composition. A pharmaceutical composition is provided, which comprises Akkermansia (Akkermansia muciniphila) and metformin, wherein the viable count of the Akkermansia is 10 based on the mass of the pharmaceutical composition6‑1014CFU/g, the content of the metformin is 20-60%.
Description
Technical Field
The present invention relates to a composition of akkermansia, more specifically to a composition of akkermansia for preventing and treating metabolic diseases.
Background
Obesity is a worldwide health problem and epidemiology has been progressively worsened by Global obesity as shown by The Global Burden of disease studies, which adversely affect The physiological hygiene and The health economics. The obesity degree is judged by taking Body Mass Index (BMI) as a standard internationally, the normal range is that the BMI is more than or equal to 18.5 and less than or equal to 24.9, the overweight is judged when the BMI is more than or equal to 25, and the obesity is judged when the BMI is more than or equal to 30. The data from 195 countries worldwide has shown that the incidence of obesity has doubled since 1980, and 5% of children and 12% of adults are currently suffering from obesity, which is similar to type ii diabetes, and which is about 1/5 people worldwide in 2025. In 1975-2014, the average BMI of men worldwide increased from 21.7 to 24.2kg/m2The female increases from 22.1 to 24.4kg/m2. By 2014, the Chinese adult obesity rate is 12.9%, the overweight rate is 41.2%, and the central obesity rate is 20.9%, and tends to increase year by year. Obesity is type 2 sugar, which destroys the metabolic homeostasis of the body and causes metabolic diseasesAn important risk factor for the onset of urine disease. Current treatments are primarily directed to reducing caloric intake and hindering caloric absorption, surgical reduction of stomach volume can reduce energy intake, basal metabolism and exercise can increase caloric expenditure and loss, and an increase in brown beige adipose tissue can increase thermogenesis. In addition, intestinal cells and flora participate in feeding behavior and can be used as a potential target for treating obesity.
Obesity and type 2 diabetes are affected by environmental and genetic factors, and the genes of human flora also play an important role in the two diseases; the flora may affect obesity by affecting the absorption of calories, etc., and obesity may in turn affect the composition of the intestinal flora. There are over 1000 different species of bacteria in the human gut, totaling about 1014. Intestinal dysbacteriosis is associated with many diseases including obesity, and the anti-inflammatory disease Akkermansia muciniphila (hereinafter, referred to as "Ackermanella") is decreased in obese patients, while Proteobacteria, Bacteroides, Campylobacter and Shigella are increased. The ratio of firmicutes to bacteroidetes not only affects carbohydrate metabolism but also alters the production of short chain fatty acids, and increased levels of acetic acid in the blood can lead to increased insulin resistance and ghrelin production, of which flora may be the most important variable factor. The intestinal flora can change various metabolic pathways centering on energy metabolism balance, thereby influencing the occurrence and development of obesity. The probiotics and prebiotics can play a role in resisting obesity by reducing intestinal bacteria lipopolysaccharide, changing flora composition, reducing fat storage and the like.
The chinese invention patent application with application No. CN201380070847.0 and publication No. CN 104918626 a discloses "use of akkermansia for treating metabolic disorders", and the akkermansia strain ATCCBAA835 screened by the patent application can be used for promoting weight loss of a subject in need thereof. However, the inventors have shown through studies that, although the akkermansia strain ATCC BAA835 is recognized as the most classical and most effective akkermansia strain, the effect of this strain in fermentation, storage stability, acid and bile salt resistance and treatment of metabolic diseases is not particularly desirable.
Metformin is a classic drug for treating type 2 diabetes and can reduce complications and mortality of type 2 diabetes, and a single dosage form of metformin has limitations in preventing and treating diseases and enhancing health. The research and study on the current situation of the Chinese type-2 diabetes oral drug treatment mode show that the most clinically applied oral hypoglycemic drug scheme in China is the combination of two drugs (45.4%), the single drug treatment (35.8%) and the small proportion of triple drug (17.0%), and most patients need the combination treatment of multiple drugs to achieve the blood sugar standard, so the research, development and preparation of the metformin compound preparation are particularly important. Currently, the metformin compound preparation mainly comprises a dipeptidyl peptidase DPP-4 inhibitor metformin compound preparation (sitagliptin metformin, saxagliptin metformin, linagliptin metformin) and a sodium glucose coordinated transporter SGLT-2 inhibitor metformin compound preparation (dapagliflozin metformin, pioglitazone metformin, engagliflozin metformin, canagliflozin metformin tablets and acarbose metformin). However, the combined treatment of metabolic diseases by using multiple drugs has the possibility of overlapping adverse effects of the drugs, and the risk of occurrence of hypoglycemic events and blood sugar drift is high. Therefore, the biological agent with mild and safe effect and the metformin are used together to form a compound preparation for the treatment of metabolic diseases. However, no biological agent and metformin compound preparation is reported at present.
There is a great need in the art for a pharmaceutical composition that can be more effectively used for the prevention and treatment of metabolic diseases.
Disclosure of Invention
The invention aims to solve the technical problems of side effect superposition, high risk of hypoglycemic events and blood sugar drift in the compound preparation of the metformin in the prior art, and provides a compound preparation for preventing and treating metabolic diseases and a preparation method thereof. The Ackermanella melbine compound preparation prepared by the invention has better effect of preventing and treating metabolic diseases, and has higher safety compared with the existing melbine compound preparation.
In one aspect, the present application provides a pharmaceutical composition comprising Akkermansia (Akkermansia muciniphila) and dimehypoGuanidine, the viable count of the strain is 10 based on the mass of the pharmaceutical composition6-1012CFU/g, the content of the metformin is 20-60%.
In a preferred embodiment of the present application, the Akkermansia akkermansoni strain Akkermansonia SSYD-3 has a collection number of CGMCC No. 14764.
In a preferred embodiment of the present application, the pharmaceutical composition comprises akkermansia and metformin as synergistic components.
In a preferred embodiment of the present application, the akkermansia is in the form of an akkermansia freeze-dried powder.
In a preferred embodiment of the present application, the pharmaceutical composition has a viable count of 10 on a mass basis8-1012CFU/g, more preferably 109-1011CFU/g, most preferably 1010CFU/g。
In a preferred embodiment of the present application, the metformin is present in an amount of 30 to 60%, more preferably 40 to 60%, still more preferably 45 to 55%, most preferably 50%.
In a preferred embodiment of the present application, the metformin is in the form of a metformin granule.
In another aspect, the present application provides the use of a combination of akkermansia and metformin for the preparation of a medicament for the prevention and treatment of a metabolic disease, or the use of a combination of akkermansia and metformin for the prevention and treatment of a metabolic disease.
In a preferred example of the present application, the use comprises reducing body weight, and/or improving glucose/insulin tolerance in a human or animal.
In a preferred embodiment of the present application, the combination of akkermansia and metformin acts as a synergistic combination.
Drawings
Fig. 1 shows the comparative effect of the present application in reducing body weight in example 1.
Fig. 2 is a comparative effect in improving glucose tolerance of effect example 1 of the present application.
Fig. 3 is a comparative effect in improving insulin tolerance of effect example 1 of the present application.
Detailed Description
In the present invention, metabolic disorders have the meaning conventionally used in the art and include diseases such as diabetes, obesity, insulin resistance, alcoholic and non-alcoholic fatty liver disease, hypertension, hyperlipidemia, hypercholesterolemia, heart disease, and the like. The prevention and treatment effects refer to prevention of occurrence and development of metabolic diseases, treatment and adjuvant therapy of metabolic diseases.
In the present invention, the percentage (%) or parts refers to the weight percentage or parts by weight with respect to the composition, unless otherwise specified.
In the present invention, the respective components referred to or the preferred components thereof may be combined with each other to form a novel embodiment, if not specifically stated.
In the present invention, all embodiments and preferred embodiments mentioned herein may be combined with each other to form a new technical solution, if not specifically stated.
In the present invention, all the technical features mentioned herein and preferred features may be combined with each other to form a new technical solution, if not specifically stated.
In the present invention, the sum of the contents of the components in the composition is 100% if not indicated to the contrary.
In the present invention, the sum of the parts of the components in the composition may be 100 parts by weight, if not indicated to the contrary.
In the present invention, unless otherwise stated, the numerical range "a-b" represents a shorthand representation of any combination of real numbers between a and b, where a and b are both real numbers. For example, a numerical range of "0 to 5" indicates that all real numbers between "0 to 5" have been listed herein, and "0 to 5" is only a shorthand representation of the combination of these numbers.
In the present invention, unless otherwise indicated, the integer numerical range "a-b" represents a shorthand representation of any combination of integers between a and b, where a and b are both integers. For example, an integer numerical range of "1-N" means 1, 2 … … N, where N is an integer.
In the present invention, unless otherwise specified, "combinations thereof" mean multicomponent mixtures of the elements described, for example two, three, four and up to the maximum possible.
The term "a" or "an" as used herein means "at least one" if not otherwise specified.
All percentages (including weight percentages) stated herein are based on the total weight of the composition, unless otherwise specified.
The "ranges" disclosed herein are in the form of lower and upper limits. There may be one or more lower limits, and one or more upper limits, respectively. The given range is defined by the selection of a lower limit and an upper limit. The selected lower and upper limits define the boundaries of the particular range. All ranges that can be defined in this manner are inclusive and combinable, i.e., any lower limit can be combined with any upper limit to form a range. For example, ranges of 60-120 and 80-110 are listed for particular parameters, with the understanding that ranges of 60-110 and 80-120 are also contemplated. Furthermore, if the minimum range values 1 and 2 are listed, and if the maximum range values 3, 4, and 5 are listed, the following ranges are all contemplated: 1-3, 1-4, 1-5, 2-3, 2-4, and 2-5.
Herein, unless otherwise specified, the proportions or weights of the components are referred to as dry weights.
In this context, "constant" means within. + -. 10%, preferably within. + -. 5%, more preferably within. + -. 2% and finally within. + -. 1% unless otherwise specified.
In this context, akkermansia includes active ingredients derived from akmansia, or bacteria that have sequence similarity of at least 97% to that of akmansia as determined by sequencing.
Herein, the term "akkermansia" or "akkermansia strain" has the same meaning unless otherwise specified.
In one aspect, the present application provides a pharmaceutical composition comprising Akkermansia (Akkermansia muciniphila) and dimehypoGuanidine, the viable count of the strain is 10 based on the mass of the pharmaceutical composition6-1014CFU/g, the content of the metformin is 20-60%.
Ackermanella
In the present invention, the akkermansia refers to a living organism or a pasteurized inactivated organism.
The akkermansia strain described herein is commercially available or can be obtained by the prior art. For example, chinese patent application No. CN201380070847.0, publication No. CN 104918626 a, discloses "use of akkermansia for treating metabolic disorders" which discloses akkermansia strain ATCC BAA 835.
In a preferred embodiment of the present application, the Ackermanella (Akkermansia muciniphila) strain is named Akkermansia muciniphila SSYD-3 with a collection number of CGMCC No. 14764. The bacterium akkermansia can be found in chinese patent application CN201811132882.1, which is incorporated herein by reference in its entirety.
The Akkermansia muciniphila SSYD-3 (W03 strain for short) is preserved in China general microbiological culture Collection center (CGMCC) in 29 th 09 th month in 2017, and the preservation address is as follows: west road No.1, north chen, chaoyang district, beijing, zip code: 100101, the preservation numbers are respectively: CGMCC No.14764, culture name Akkermansia muciniphila SSYD-3, and classification name Akkermansia muciniphila.
The akkermansia can be prepared in various suitable forms, such as solutions, tablets, capsules, orally disintegrating tablets, lyophilized powders, and the like. In a preferred embodiment of the present application, the form is a lyophilized powder. In another preferred embodiment of the present application, the dosage form is a tablet. The dosage form is preferably a lyophilized powder. The viable count of the Ackermanella is preferably 1010CFU/g. Wherein the effective dose refers to that the total viable count of the solid pharmaceutical composition prepared by taking Ackermanella as a main pharmaceutical active ingredient is 106-1014CFU/g. Wherein the effective dose is a liquid prepared by taking Ackermanella as a main medicinal active ingredientThe total viable count of the pharmaceutical composition is 106-1014CFU/mL。
In the present invention, the akkermansia bacteria is preferably prepared as a pharmaceutical composition in the form of an akkermansia freeze-dried powder.
The preparation method of the freeze-dried powder of the Ackermanella preferably comprises the following steps:
and (3) uniformly mixing the centrifugal thallus of the Ackermanella with a freeze-drying protective agent and water, and freeze-drying to obtain the product.
Among them, the centrifuged cells of Ackermanella are preferably obtained as follows: and centrifuging the fermentation liquor of the Ackermanella, and collecting thalli. The centrifugation is a routine operation in the art, for example centrifugation at 8000rpm for 20 min.
Wherein, the viable count of the Acermann's bacteria in the Acermann's bacteria freeze-dried powder is preferably 10 based on the mass of the pharmaceutical composition6-1014CFU/g, more preferably 1010CFU/g。
Wherein, the raw material of the lyoprotectant is the conventional lyoprotectant raw material in the field. For example, in a preferred embodiment of the present application, the lyoprotectant is a mixture of sodium glutamate, mannitol, trehalose, and milk powder; in the preparation step of the akkermansia freeze-dried powder, the mass percentages of sodium glutamate, mannitol, trehalose and milk powder in the freeze-dried liquid to be obtained by uniform mixing are respectively 2%, 2%, 4% and 8%.
The freeze-dried powder of the Ackermanella can be crushed, sieved by a 40-mesh sieve and stored in a turnover barrel for sealing and storage for later use according to the conventional method in the field.
Metformin
In the present application, the term "metformin" means metformin or a derivative thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable ester thereof, or a prodrug thereof.
In the present application, the metformin is preferably prepared in the form of metformin granules.
The preparation method of the metformin hydrochloride granules is conventional in the field, and preferably comprises the following steps: mixing metformin and binder uniformly, granulating by wet method, drying, grading, and mixing with lubricant uniformly.
Among them, metformin is usually used as a raw material for medicines in the form of hydrochloride, sulfate, etc., i.e., metformin hydrochloride, metformin sulfate, etc. Metformin hydrochloride and metformin sulfate are preferably further subjected to pulverization treatment and then sieved with a 40-mesh sieve.
Wherein the binder is conventional in the art, such as povidone K30.
Wherein the lubricant is a substance conventional in the art, such as magnesium stearate.
Wherein the metformin granules can be stored in a turnover barrel for sealed storage for later use according to the conventional method in the field.
Pharmaceutical composition
The pharmaceutical composition can be prepared into any suitable dosage form, such as oral liquid, tablet, capsule, orally disintegrating tablet, injection, etc., by adding pharmaceutically acceptable carriers. The above pharmaceutical composition can be easily prepared into the above dosage forms by those skilled in the art according to the prior art.
In one embodiment, the present application further provides a method for preparing the above pharmaceutical composition, comprising the following steps: the raw materials of the pharmaceutical composition are uniformly mixed.
In one example of the present application, the viable count is 10 based on the mass of the pharmaceutical composition6-1014CFU/g, preferably 108-1012CFU/g, more preferably 109-1011CFU/g, most preferably 1010CFU/g。
In another preferred embodiment of the present application, the metformin is present in an amount of 20 to 60%, preferably 30 to 60%, more preferably 40 to 60%, still more preferably 45 to 55%, most preferably 50% by mass of the pharmaceutical composition.
In one example of the present application, the pharmaceutical composition comprises akkermansia and metformin as synergistic components.
In the invention, when the Acermanniella and the metformin respectively take the forms of the Acermanniella freeze-dried powder and the metformin granules to participate in the preparation of the compound preparation, the mass ratio of the Acermanniella freeze-dried powder to the metformin granules is preferably 1: 2-1: 10.
According to a preferred embodiment of the invention, the compound preparation is a tablet, which is obtained by the following method: mixing and tabletting the freeze-dried powder of the Acermanniella anatipestifer and the metformin granules according to the mass ratio of 1: 10.
Wherein, during tabletting, the thickness of the tabletting is controlled to be the conventional thickness, such as 5.0 mm-5.5 mm.
According to another preferred embodiment of the invention, the compound preparation is an enteric capsule, which is obtained by the following method: uniformly mixing the akkermansia freeze-dried powder, the metformin granules and the carboxymethyl cellulose in a mass ratio of 1:2:2, and filling into capsules.
Wherein the mixing is carried out under conventional operating conditions, such as 8rpm for 30 min.
Wherein the capsule is an enteric capsule commonly used in the field.
On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.
The reagents and starting materials used in the present invention are commercially available.
The positive progress effects of the invention are as follows: the Ackermanella has a prominent effect on reducing blood sugar, is human intestinal resident bacteria, and ensures the safety of symbiosis with a host, and the metformin is a first-line medicament for reducing blood sugar in type 2 diabetes.
A further aspect of the application provides the use of a combination of akkermansia and metformin for the preparation of a medicament for the prevention and treatment of metabolic diseases.
A further aspect of the present application provides the use of a combination of akkermansia and metformin for the prevention and treatment of metabolic diseases.
In one example of the present application, the viable count is 10 based on the mass of the pharmaceutical composition6-1012CFU/g, preferably 108-1012CFU/g, more preferably 109-1011CFU/g, most preferably 1010CFU/g。
In another preferred embodiment of the present application, the metformin is present in an amount of 20 to 60%, preferably 30 to 60%, more preferably 40 to 60%, still more preferably 45 to 55%, most preferably 50% by mass of the pharmaceutical composition.
In a preferred example of the present application, the use comprises reducing body weight, and/or improving glucose/insulin tolerance in a human or animal.
The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.
Example 1: preparation of Ackermanella melbine compound tablet
(1) Preparation of W03 lyophilized powder
① freeze-drying protectant including sodium glutamate, mannitol, trehalose, and milk powder.
② the fermentation broth of the W03 bacterium is centrifuged for 20min at 8000rpm, and the bacterium is collected.
③ centrifuging Achromax mansoni to obtain thallus, mixing with freeze-drying protective agent and water to make the mass percentage of sodium glutamate, mannitol, trehalose and milk powder in the solution before freeze-drying respectively be 2%, 2%, 4% and 8%, freeze-drying to obtain freeze-dried thallus powder, wherein the viable count of the Achromax mansoni strain with preservation number of CGMCC No.14764 in the thallus powder is 1010CFU/g. The obtained W03 bacterial powder is crushed and stored in a turnover barrel to be sealed and stored for later use.
(2) Preparation of metformin granules
Metformin hydrochloride (10 kg, Shandong Shouguangkukang pharmaceutical Co., Ltd.) was pulverized and sieved with a 40-mesh sieve to obtain metformin hydrochloride, and mixed with povidone K30 (10 kg) as a binder, followed by wet granulation and drying. And (3) putting the dry particles into a lifting granulator for dry particle granulation, and transferring the granulated particles into a mixing box. Mixing the granules with magnesium stearate, adding into a mixing machine, mixing for 10min, and discharging. The obtained metformin granules are stored in a turnover barrel for sealed preservation and standby.
(3) Tabletting
Mixing and tabletting W03 bacterial powder and metformin granules in a mass ratio of 1: 10.
A beveled circular punch was used. And controlling the thickness of the tablet to be 5.0-5.5 mm during tabletting, and sealing and storing the pressed Ackermansia melbine compound tablet after tabletting.
Example 2: preparation of compound enteric capsule of Acermanniella mellea and metformin
(1) Preparation of W03 lyophilized powder
① freeze-drying protectant including sodium glutamate, mannitol, trehalose, and milk powder.
② W03 bacteria fermented fermentation broth 8000rpm, centrifuging for 20min, and collecting bacteria.
③ centrifuging Achromax mansoni to obtain thallus, mixing with freeze-drying protective agent and water to make the mass percentage of sodium glutamate, mannitol, trehalose and milk powder in the solution before freeze-drying respectively be 2%, 2%, 4% and 8%, freeze-drying to obtain freeze-dried thallus powder, wherein the viable count of the Achromax mansoni strain with preservation number of CGMCC No.14764 in the thallus powder is 1010CFU/g. The obtained W03 bacterial powder is crushed and stored in a turnover barrel to be sealed and stored for later use.
(2) Preparation of metformin granules
Metformin hydrochloride (10 kg, Shandong Shouguangkukang pharmaceutical Co., Ltd.) was pulverized and sieved with a 40-mesh sieve, and the obtained metformin hydrochloride was mixed with 10kg of povidone K30 as a binder, followed by wet granulation and drying. And (3) putting the dry particles into a lifting granulator for dry particle granulation, and transferring the granulated particles into a mixing box. Mixing the granules with magnesium stearate, adding into a mixing machine, mixing for 10min, and discharging. And storing the metformin granules in a turnover barrel for sealing and storing for later use.
(3) Capsule preparation
According to the mass ratio of W03 bacterial powder: metformin granules: adding the carboxymethyl cellulose at a ratio of 1:2:2 into a charging basket, transferring the mixture into a revolving body frame of a mixer at a rotating speed of 8rpm, mixing for 30 minutes, and filling the mixed material into enteric capsules.
Effect example 1: application of Ackermanella and metformin compound preparation in treatment of metabolic diseases
Male C57 BL/6 mice (no specific pathogen, shanghai laboratory animal center) at 8 weeks of age were fed food freely in groups of 3 mice per cage (filter cage) during a strict 12-hour light cycle. After they were acclimatized, they were fed either a standard Diet or a high fat Diet with 60 kcal% fat for 12 consecutive weeks (Research Diet, D12492 i). After 12 weeks of high-fat diet dry prognosis, animals were randomly divided into 4 groups, and 200. mu.l of PBS (PBS group) and 200. mu.l of Ackermanella (0.1 g of 10 bacteria content)10CFU/g of akkermansia (akkermansia strain with a collection number of CGMCC No. 14764) powder was dissolved in 1ml of pbs solution, designated as W03 group), 200 μ l of metformin hydrochloride single-use group (prepared by dissolving 0.3g of metformin hydrochloride from shandong shouguang fukang pharmaceutical co., ltd., identified as MET group) and 200 μ l of a mixed solution of akkermansia and metformin (the mixed solution was prepared as follows: 0.3g of metformin, 0.1g of bacteria content of 1010CFU/g of Arkermanella bacteria powder was dissolved in 1ml PBS solution, designated C03 or CW 03), 1 time per day for 8 weeks.
Body weight was measured before gavage and at the end of the experiment. The amount of food ingested for 7 consecutive days was recorded in cages (3 mice per cage) and the average food intake per mouse per day was calculated. Stool samples were collected before gavage and at the end of the experiment and immediately stored in a-80 ℃ freezer for further analysis. Whole blood was taken from the tail vein of the mouse, and its blood glucose level and insulin level were measured with a glucometer (LifeScan). To ensure the same starting point, the study did not use a random distribution method, but rather the environmentally adapted mice were grouped according to body weight. All procedures were approved by the animal ethics committee of the Shanghai university of transportation medical college.
(1) The compound preparation can remarkably reduce the weight
Compared with the Ackermanella group and the metformin group in the control group, the weight of the Ackermanella-metformin compound combination group is obviously reduced. As shown in particular in figure 1. FIG. 1 shows the results of the test at the end of week 8 of the experiment, and the data are shown in Table 1.
TABLE 1 weight gain in the test groups
| Test group number | Percentage of body weight gain |
| PBS group | 24.54％ |
| MET group | 4.46％ |
| W03 group | 6.94％ |
| Group CW03 | -1.27％ |
(2) The compound preparation can remarkably improve the glucose and insulin tolerance
The test result shows that: compared with the Ackermanella group and the metformin group in the control group, the compound combination group of the Ackermanella and the metformin has obviously improved glucose and insulin tolerance and embodies a synergistic cooperation effect. The results of the glucose tolerance test are specifically shown in fig. 2. Figure 2 is the results of blood glucose testing over 2 hours after glucose injection at the end of week 8 of the experiment and the data are shown in tables 2-5.
TABLE 2 glucose values data for the test groups
| |
0 |
15 |
30 |
60 |
120 minutes |
| PBS group | 9.24 | 17.41 | 15.79 | 11.87 | 8.29 |
| MET group | 7.28 | 16.40 | 16.25 | 11.63 | 7.75 |
| W03 group | 6.34 | 12.76 | 11.50 | 8.40 | 5.89 |
| Group C03 | 5.53 | 12.79 | 10.56 | 7.03 | 5.36 |
TABLE 3 statistical data (P-value) of glucose blood sugar values between test groups
Data in the table with a P value of 0.05 or less are shown to have statistical differences.
TABLE 4 area under 0-120 min glucose tolerance curve for each test group
| Test group number | AUC |
| PBS group | 1468.5 |
| MET group | 1422.2 |
| W03 group | 1052.4 |
| Group C03 | 947.8 |
TABLE 5 statistical data (P-value) of the area under the 0-120 min glucose tolerance curve between test groups
| Test group number | P value |
| MET vs PBS | 0.61195 |
| W03 vs PBS | 0.00004 |
| C03 vs PBS | 0.00010 |
| C03 vs W03 | 0.12861 |
| C03 vs MET | 0.00003 |
Data in the table with a P value of 0.05 or less are shown to have statistical differences.
The results of the insulin tolerance test are shown in detail in fig. 3. Figure 3 is the results of blood glucose measurements taken at the end of week 8 and within 2 hours of insulin injection, and the data are shown in tables 6-9.
TABLE 6 glucose values data for the test groups
| |
0 |
15 |
30 |
60 |
120 minutes |
| PBS group | 10.09 | 9.31 | 7.57 | 6.74 | 7.64 |
| MET group | 8.54 | 8.01 | 6.29 | 5.79 | 6.66 |
| W03 group | 10.29 | 9.05 | 7.40 | 6.74 | 7.90 |
| Group C03 | 8.84 | 7.24 | 4.90 | 3.91 | 4.07 |
TABLE 7 statistical data (P-value) of glucose blood sugar values between test groups
Data in the table with a P value of 0.05 or less are shown to have statistical differences.
TABLE 8 area under 0-120 min glucose tolerance curve for each test group
| Test group number | AUC |
| PBS group | 918.4 |
| MET group | 786.1 |
| W03 group | 919.6 |
| Group C03 | 583.5 |
TABLE 9 statistical data (P-value) of the area under the 0-120 min glucose tolerance curve between test groups
| Test group number | P value |
| MET vs PBS | 0.15517 |
| W03 vs PBS | 0.98991 |
| C03 vs PBS | 0.00052 |
| C03 vs W03 | 0.00029 |
| C03 vs MET | 0.01548 |
Data in the table with a P value of 0.05 or less are shown to have statistical differences.
In fig. 2 and 3, the W03 group had an effect on glucose tolerance and had no effect on pancreatic islet tolerance for the following reasons: the improvement of sugar metabolism is achieved by a number of mechanisms. Mainly comprises reducing the source of glucose and increasing the outward route of the glucose. It is found through experiments that W03 can improve glucose tolerance, probably mainly by increasing the utilization of glucose by peripheral tissues and organs, reducing hepatic glucose output and the like. W03 did not significantly improve insulin tolerance, suggesting that W03 application did not increase the sensitivity of the subject to exogenous insulin, and that its effect on improving glucose metabolism was not achieved by increasing insulin sensitivity.
Claims (10)
1. A pharmaceutical composition comprising Akkermansia (Akkermansia mu)ciniphila) and metformin, the viable count of the Ackermanella is 10 based on the mass of the pharmaceutical composition6-1014CFU/g, the content of the metformin is 20-60%.
2. The pharmaceutical composition according to claim 1, wherein the akkermansia akkermansonia strain SSYD-3 has a collection number of CGMCC No. 14764.
3. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises akkermansia and metformin as synergistic components.
4. The pharmaceutical composition of claim 1, wherein said akkermansia is in the form of an akkermansia lyophilized powder.
5. The pharmaceutical composition of claim 1, wherein the viable count is 10 based on the mass of the pharmaceutical composition8-1012CFU/g, more preferably 109-1011CFU/g, most preferably 1010CFU/g。
6. A pharmaceutical composition according to claim 1, wherein metformin is present in an amount of 30-60%, more preferably 40-60%, still more preferably 45-55%, most preferably 50%.
7. The combination of claim 1, wherein the metformin is in the form of metformin granules.
8. Use of a combination of akkermansia and metformin for the preparation of a medicament for the prevention and treatment of a metabolic disease, or use of a combination of akkermansia and metformin for the prevention and treatment of a metabolic disease.
9. The use according to claim 8, wherein the use comprises reducing body weight, and/or improving glucose/insulin tolerance in a human or animal.
10. The use according to claim 8, wherein the combination of akkermansia and metformin is used as a synergistic combination.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010096535.9A CN111202752A (en) | 2020-02-17 | 2020-02-17 | Ackermanella compositions |
| PCT/CN2021/074307 WO2021164523A1 (en) | 2020-02-17 | 2021-01-29 | Akkermansia muciniphila composition |
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| CN202010096535.9A CN111202752A (en) | 2020-02-17 | 2020-02-17 | Ackermanella compositions |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021164523A1 (en) * | 2020-02-17 | 2021-08-26 | 上海市内分泌代谢病研究所 | Akkermansia muciniphila composition |
| CN113322202A (en) * | 2021-05-31 | 2021-08-31 | 君维安(武汉)生命科技有限公司 | Ackermanella, culture method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104918626A (en) * | 2012-11-19 | 2015-09-16 | 卢万天主教大学 | Use of akkermansia for treating metabolic disorders |
| CN108778309A (en) * | 2016-01-26 | 2018-11-09 | 防御素治疗学公司 | The method for adjusting intestinal microbiota |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| EP3359171B1 (en) * | 2015-10-05 | 2023-07-05 | Schweizerisches Forschungsinstitut für Hochgebirgsklima und Medizin in Davos | Use of akkermansia muciniphila for treating inflammatory conditions |
| CN111202752A (en) * | 2020-02-17 | 2020-05-29 | 上海市内分泌代谢病研究所 | Ackermanella compositions |
-
2020
- 2020-02-17 CN CN202010096535.9A patent/CN111202752A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104918626A (en) * | 2012-11-19 | 2015-09-16 | 卢万天主教大学 | Use of akkermansia for treating metabolic disorders |
| CN108778309A (en) * | 2016-01-26 | 2018-11-09 | 防御素治疗学公司 | The method for adjusting intestinal microbiota |
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| 沈男等: "嗜黏蛋白阿克曼氏菌及其在肥胖机制中的研究进展", 《基础医学与临床》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021164523A1 (en) * | 2020-02-17 | 2021-08-26 | 上海市内分泌代谢病研究所 | Akkermansia muciniphila composition |
| CN113322202A (en) * | 2021-05-31 | 2021-08-31 | 君维安(武汉)生命科技有限公司 | Ackermanella, culture method and application thereof |
| CN113322202B (en) * | 2021-05-31 | 2022-03-01 | 君维安(武汉)生命科技有限公司 | Ackermanella, culture method and application thereof |
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