RU2806833C2 - Composition containing a mixture of plant extracts or a mixture of molecules contained in the specified plants and use for controlling the metabolism of carbohydrates and/or lipids - Google Patents

Abstract

FIELD: pharmaceutical industry.

SUBSTANCE: group of inventions relates to a composition and a method of prevention or treatment of carbohydrate and/or fat metabolism disorders in humans or animals. A composition for the prevention or treatment of carbohydrate and/or fat metabolism disorders in humans or animals, containing at least a mixture of molecules containing at least the following: a single extract obtained from a mixture of at least two plants selected from Chrysanthellum indicum, Cynara scolymus, Vaccinium myrtillus, Olea europaea and Piper, and where the mixture of molecules further contains at least one of the following extracts: one Chrysanthellum indicum extract, if the plant mixture of a single extract does not contain Chrysanthellum indicum; one extract of Cynara scolymus, if the plant mixture of a single extract does not contain Cynara scolymus; one extract of Vaccinium myrtillus, if the plant mixture of the single extract does not contain Vaccinium myrtillus; piperine or Piper extract if the single extract plant mixture does not contain Piper; and/or one Olea europaea extract, if the mixture of plants of a single extract does not contain Olea europaea, where the specified mixture of molecules contains the following: at least one of apigenin-7-O-glucuronide, chrysanthellin A, chrysantellin B, caffeic acid, luteolin, maritimethine, eriodictyol, isoocanin, apigenin, luteolin-7-O-glucoside, maritimein, marein, eriodictyol-7-O-glucoside, flavomarein, apigenin-8-C-α -L-arabinoside-6-C-β-D-glucoside (shaftoside), apigenin-6,8-C-di-β-D-glucopyranoside (vicenin-2), and at least one of the following: dicaffeoylquinic acid, sulfomonocaffeoylquinic acid, luteolin, luteolin-7-O-glucoside, luteolin-7-O-glucuronide, apigenin-7-O-glucoside, cinaropicrin, and at least one of monocaffeylquinic acid, delphinidin-3-galactoside, delphinidin-3-glucoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, cyanidin-3-glucoside, petunidin-3-galactoside, cyanidin-3-arabinoside , petunidin-3-glucoside, peonidin-3-galactoside, petunidin-3-arabinoside, peonidin-3-glucoside, malvidin-3-galactoside, malvidin-3-glucoside, malvidin-3-arabinoside, and at least piperine, and at least one of oleuropein, hydroxytyrosol; where: a single extract or extract of Chrysanthellum indicum is obtained using the whole plant and/or aerial parts of Chrysanthellum indicum; a single extract or extract of Cynara scolymus is obtained using the whole plant and/or leaves of Cynara scolymus; a single extract or extract of Vaccinium myrtillus is obtained using the whole plant and/or fruit of Vaccinium myrtillus; a single extract or extract of Olea europaea is obtained using the fruits or leaves of Olea europaea; a single extract or Piper extract is prepared using the fruits or leaves of Piper nigrum, Piper aduncum and/or Piper longum; wherein the extracts of Chrysanthellum indicum, Vaccinium myrtillus, Olea europaea and Piper are obtained using a water/ethanol mixture, the extract of Cynara scolymus is obtained using water; where the composition contains such an amount of the corresponding extract that allows the introduction of at least 0.00001 g of Chrysanthellum indicum extract, at least 0.00001 g of Cynara scolymus extract, at least 0.00001 g of Vaccinium myrtillus extract, at least 0.001 mg of piperine and at least 0.00001 g of Olea europaea extract per kg of body weight of the person to whom the composition is administered, per day. A method of prevention or treatment of pathological disorders of carbohydrate and/or fat metabolism in humans or animals, including administering the above-described composition to a person or animal as a food product and/or medicine.

EFFECT: above-described composition and the method of its use exhibit synergism in the prevention or treatment of pathological disorders of carbohydrate and/or fat metabolism in humans or animals.

20 cl, 12 dwg, 17 tbl, 15 ex

Description

The present invention relates to the prevention and treatment of carbohydrate and/or fat metabolism disorders in humans and animals.

Type 2 diabetes, the most common form of diabetes, is a chronic, progressive metabolic disease. It is characterized by chronic hyperglycemia, i.e. pathologically high concentration of sugar in the blood, and intolerance to carbohydrates. The main cause of this chronic hyperglycemic state is insulin resistance and inadequate insulin secretion in response to this metabolic state, but other factors may be involved (Ismail-Beigi FN Engl J Med 2012; 366:1319-27). The main approaches to control type 2 diabetes mellitus should be to reduce glycemic levels, reduce glycated hemoglobin (HbA _1c ) levels to less than or equal to 6.5%, and improve insulin sensitivity.

Treatment of type 2 diabetes mellitus consists primarily of lifestyle changes or compliance with hygiene and dietary measures (HDM: diet, smoking cessation, physical activity and sports activities). If HDM is not enough, antidiabetic therapeutic agents, mainly metformin, should be used. However, metformin has many medical contraindications, such as chronic renal failure, acidosis, hypoxia, dehydration, etc. Thus, the use of metformin is essentially a paradox, since it cannot be prescribed in patients with type 2 diabetes mellitus who have renal failure, despite the fact that renal failure is one of the common consequences of type 2 diabetes mellitus. Other therapeutic agents have been developed, such as dipeptidyl peptidase-IV (DPP-IV) inhibitors, glucagon-like peptide-1 (GLP-1) analogues, thiazolidinediones (TZDs), sulfonylureas, glycosidase inhibitors (acarbose, miglitol, voglibose) or sodium-dependent cotransporter inhibitors glucose-2 (SGLT2). However, designing the right therapeutic combination is difficult because a large number of factors must be taken into account, such as contraindications and adverse reactions, which affect patients' quality of life and therefore their compliance with treatment. In addition, some therapeutic combinations increase overall mortality, such as the combination of metformin with sulfonylureas (Prescrire Int 2015;24:130-5).

Since patients very rarely adhere to HDM, it is necessary to very quickly initiate therapeutic treatment, taking into account all the adverse reactions and contraindications associated with these molecules, and currently a solution suitable for helping patients with type 2 diabetes between HDM and the initiation of therapeutic treatment absent.

In addition, patients with type 2 diabetes are at high risk of morbidity and mortality from cardiovascular disease. Thus, well-known cardiovascular risk factors, such as control of circulating fat, body weight and blood pressure, also need to be included in patient care. This need currently involves the simultaneous use of several drugs of different therapeutic classes. However, combining drugs can in some cases cause serious adverse reactions, for example, concomitant use of fibrates and statins increases the risk of myopathy (Denke MA J Manag Care Pharm 2003; 9:17-9).

Therefore, there is a real need for drugs that can be used both during the gradual development of cardio-metabolic pathologies, characterized by an increase in certain risk factors (metabolic disorders of carbohydrates, fats, excess body weight, inflammation, oxidative stress, arterial hypertension), and during an outbreak these pathologies, especially type 2 diabetes mellitus.

In addition, there is an urgent need for preventive measures and drugs whose “multi-target” mechanism of action has advantages in terms of suitability, tolerability and efficacy. Such drugs may make it possible to reduce the overall risk of cardio-metabolic diseases, as well as prevent and treat each dysfunction and/or its consequences separately.

The purpose of the invention is to meet these various needs by providing compositions capable of simultaneously addressing multiple dysfunctions of carbohydrate and fat metabolism, representing both prophylactic and therapeutic agents with benefits for the prevention and treatment of cardio-metabolic diseases and their complications.

To achieve this goal, the invention relates to compositions containing at least a mixture of molecules obtained from at least:

- Chrysanthellum indicum, and

- Cynara scolymus, and

- Vaccinium myrtillus,

wherein said mixture of molecules also contains piperine.

This mixture is a synergistic mixture because the molecules present in the mixture act synergistically.

Chrysanthellum indicum extracts, Cynara scolymus extracts, Vaccinium myrtillus extracts and piperine have already been described and some of them are used in food products, but surprisingly the combination of at least one Chrysanthellum indicum extract with at least one Cynara scolymus extract, Vaccinium myrtillus extract and piperine results to amazing results in both carbohydrate and fat metabolism in humans or animals.

To achieve this object, the invention also relates to compositions consisting of at least piperine and specific specific molecules contained in Chrysanthellum indicum, Cynara scolymus and Vaccinium myrtillus (where these molecules are natural and/or synthetic), in particular to a composition containing a mixture at least four molecules, where at least one molecule is piperine, at least one molecule selected from apigenin-7-O-glucuronide, chrysanthellin A, chrysanthellin B, caffeic acid, luteolin, maritimethine, eriodictyol, isoocanin, apigenin, luteolin-7-O-glucoside, maritimein, marein, eriodictyol-7-O-glucoside, flavomarein, apigenin-8-C-α-L-arabinoside-6-C-β-D-glucoside (shaftoside), apigenin-6 ,8-C-di-β-D-glucopyranoside (vicenin-2) or analogs thereof, and at least one molecule selected from dicaffeylquinic acid, sulfomonocaffeylquinic acid, luteolin, luteolin-7-O-glucoside, luteolin-7-O -glucuronide, apigenin-7-O-glucoside, cynaropicrin or analogs thereof, and at least one molecule selected from monocaffeylquinic acid, delphinidin-3-galactoside, delphinidin-3-glucoside, cyanidin-3-galactoside, delphinidin-3-arabinoside , cyanidin-3-glucoside, petunidin-3-galactoside, cyanidin-3-arabinoside, petunidin-3-glucoside, peonidin-3-galactoside, petunidin-3-arabinoside, peonidin-3-glucoside, malvidin-3-galactoside, malvidin -3-glucoside, malvidin-3-arabinoside or analogues thereof.

In this application, the singular or plural is used to refer to compositions according to the invention without preference.

Advantageously, the compositions according to the invention prevent the development of chronic hyperglycemia and reduce glycemia, reduce the level of glycated hemoglobin, improve tolerance to carbohydrates when taken orally, increase insulin sensitivity, but are also able to act on other risk factors for cardiovascular diseases, such as dyslipidemia, excess body weight and obesity and blood pressure. In addition, they have few or no side effects compared to existing drugs and those in development.

The invention thus also relates to the use of the compositions as food or health products, especially as medicinal products, in particular for the prevention and/or control of carbohydrate and fat metabolism disorders in humans or animals.

The invention is described in detail below with reference to the accompanying figures, which show:

- Fig. 1: results of oral insulin sensitivity test for four compositions corresponding to the results in Table 9 (item II.1);

- Fig. 2: results demonstrating the synergistic effect obtained as a result of the use of the composition according to the invention, these results correspond to the results shown in Table 13 (item II.1) for fasting glycemia;

- Fig. 3: results demonstrating the synergistic effect obtained as a result of the use of the composition according to the invention, these results correspond to the results shown in Table 13 (item II.1) for carbohydrate tolerance;

- Fig. 4: results demonstrating the synergistic effect obtained as a result of the use of the composition according to the invention, these results correspond to the results shown in Table 14 (item II.1) for fasting glycemia;

- Fig. 5: results demonstrating the synergistic effect obtained as a result of the use of the composition according to the invention, these results correspond to the results shown in Table 14 (item II.1) for carbohydrate tolerance;

- Fig. 6: results demonstrating the synergistic effect obtained as a result of the use of the composition according to the invention, these results correspond to the results shown in Table 14 (item II.1) for insulin sensitivity;

- Fig. 7: results of the effect of the composition according to the invention and metformin on insulin sensitivity, these results correspond to the results shown in Table 15 (item II.2);

- Fig. 8: results of the effect of the composition according to the invention on changes in body weight, these results correspond to the results given in Table 15 (item II.2);

- Fig. 9: the results of the effect of the composition according to the invention on changes in fasting glycemia, these results correspond to the results shown in Table 15 (item II.2);

- Fig. 10: Western blot results showing an increase in the amount of AMP-activated protein kinase in the liver of mice when using the composition according to the invention (item II.2);

- Fig. 11: test results demonstrating the absence of acute hypoglycemic effect when using the composition according to the invention (clause II.3);

- Fig. 12: test results demonstrating the effect of the composition according to the invention on changes in body weight gain (item II.4).

The subject of the invention is a composition containing at least a mixture of molecules obtained from at least:

- Chrysanthellum indicum, and

- Cynara scolymus, and

- Vaccinium myrtillus,

wherein said mixture of molecules also contains piperine.

Thus, the invention relates to a composition containing at least piperine, at least one molecule isolated from Chrysanthellum indicum, at least one molecule isolated from Cynara scolymus, and at least one molecule isolated from Vaccinium myrtillus.

In addition to Chrysanthellum indicum, Cynara scolymus, Vaccinium myrtillus and piperine, the composition according to the invention may also contain other compounds, in particular Olea europea extract. In this case, the composition according to the invention contains at least one molecule isolated from Chrysanthellum indicum, at least one molecule isolated from Cynara scolymus, at least one molecule isolated from Vaccinium myrtillus, and at least one molecule isolated from Olea europea, and piperine.

According to a first embodiment of the invention, the composition according to the invention contains at least:

- Chrysanthellum indicum extract, and

- Cynara scolymus extract, and

- Vaccinium myrtillus extract, and

- synthetic piperine and/or Piper extract containing piperine.

For the purposes of the invention, the term “extract of plant “X” or plant material “X”” means at least one molecule, preferably a group of molecules, obtained from plant “X” by any suitable method. In particular, mention may be made of aqueous extracts (obtained using an aqueous solvent), alcoholic extracts (obtained using an alcohol solvent), either obtained using an organic solvent or using a natural fatty substance or mixture of natural fatty substances, especially vegetable oil or mixture of vegetable oils . The term "aqueous solvent" means any solvent consisting wholly or partially of water. Thus, mention may be made of water itself, hydroalcoholic solvents in any proportions, or solvents consisting of water and a compound such as glycerin or propylene glycol in any proportions. Among alcohol solvents, ethanol may be particularly mentioned.

For the purposes of the invention, the term “plant or plant material” means the whole plant or part of a plant, including cell cultures that have not yet undergone special processing, and intended for inclusion in the production of a herbal preparation.

Plant extracts can be obtained by any suitable method, for example, by a method comprising the following steps:

- solid-liquid extraction

- separation/pressing

- filtering

- evaporation

- drying

- optional addition of additives

- homogenization

- air conditioning.

The Chrysanthellum indicum extract is preferably an extract of the whole plant or its aerial parts.

It may in particular be a hydroalcoholic, or aqueous, or subcritical CO ₂ extract, or a subcritical H ₂ O extract, or an extract obtained by extraction in combination with heat treatment carried out by standard heating, or at microwave frequency, or under the influence of ultrasound.

The plant/extract ratio is advantageously from 1/1 to 100/1, in particular from 1/1 to 25/1.

The composition according to the invention intended for humans preferably contains an amount of Chrysanthellum indicum extract that allows the administration of at least 0.00001 g, in particular from 0.00001 g to 0.60 g, of Chrysanthellum indicum extract per kg body weight of the person to whom the composition is administered per day.

Preferably, the Chrysanthellum indicum extract contains at least one molecule selected from apigenin-7-O-glucuronide, chrysantellin A, chrysantellin B, caffeic acid, luteolin, maritimethine, eriodictyol, isoocanin, apigenin, luteolin-7-O-glucoside, maritimein , mareina, eriodictyol-7-O-glucoside, flavomarein, apigenin-8-C-α-L-arabinoside-6-C-β-D-glucoside (shaftoside), apigenin-6,8-C-di-β- D-glucopyranoside (vicenin-2) or their analogues. Advantageously, the extract contains at least apigenin-7-O-glucuronide.

For the purposes of the present invention, the term “analogs” means all compounds having a chemical structure similar to that of another compound, but differing from it in a specific component. It may be distinguished by one or more atoms, functional groups, substructures, which are replaced by other atoms, functional groups or substructures. Examples that may be mentioned include apigenin-7-O-glucuronide analogs such as apigenin-7-apioglucoside, apigenin-8-C-glucoside (Vitexin), apigenin-6-C-glucoside (isovitexin), apigenin-7- O-neospheridoside, apigenin-7-glucoside, apigenin-7-apioglucoside.

The Cynara scolymus extract is preferably a leaf or root extract.

It may in particular be a hydroalcoholic, or aqueous, or subcritical CO ₂ extract, or a subcritical H ₂ O extract, or an extract obtained by extraction in combination with heat treatment carried out by standard heating, or at microwave frequency, or under the influence of ultrasound.

The plant/extract ratio is advantageously from 1/1 to 100/1, in particular from 1/1 to 30/1.

The composition according to the invention intended for humans advantageously contains an amount of Cynara scolymus extract that allows the administration of at least 0.00001 g, in particular from 0.00001 g to 0.60 g, of Cynara scolymus extract per kg of body weight of the person to whom the composition is administered per day.

Preferably, the Cynara scolymus extract contains at least one molecule selected from dicaffeoylquinic acid, sulfomonocaffeoylquinic acid, luteolin, luteolin-7-O-glucoside, luteolin-7-O-glucuronide, apigenin-7-O-glucoside, cinaropicrin or analogs thereof . Advantageously, the extract contains at least one dicaffeoylquinic acid.

The Vaccinium myrtillus extract is preferably a fruit or leaf extract.

It may in particular be a hydroalcoholic, or aqueous, or subcritical CO ₂ extract, or a subcritical H ₂ O extract, or an extract obtained by extraction in combination with heat treatment carried out by standard heating, or at microwave frequency, or under the influence of ultrasound.

The plant/extract ratio is advantageously from 1/1 to 200/1, in particular from 1/1 to 60/1.

The composition according to the invention intended for humans preferably contains an amount of Vaccinium myrtillus extract that allows the administration of at least 0.00001 g, in particular from 0.00001 g to 0.60 g, of Vaccinium myrtillus extract per kg of body weight of the person to whom the composition is administered per day.

Preferably, the Vaccinium myrtillus extract contains at least one molecule selected from monocaffeylquinic acid, delphinidin-3-galactoside, delphinidin-3-glucoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, cyanidin-3-glucoside, petunidin-3 -galactoside, cyanidin-3-arabinoside, petunidin-3-glucoside, peonidin-3-galactoside, petunidin-3-arabinoside, peonidin-3-glucoside, malvidin-3-galactoside, malvidin-3-glucoside, malvidin-3-arabinoside or their analogues. Advantageously, the extract contains at least one dicaffeoylquinic acid.

The piperine present in the composition of the invention may be contained in Piper extract or may be synthetic piperine.

The topological formula of piperine is given below:

The composition according to the invention intended for humans advantageously contains an amount of piperine that allows the administration of at least 0.001 mg, in particular from 0.001 mg to 166 mg, piperine per kg of body weight of the person to whom the composition is administered, per day.

If piperine is contained in the Piper extract, the mixture of the composition according to the invention contains the specified extract. Piper extract is predominantly an extract of Piper nigrum, Piper aduncum and/or Piper longum.

The Piper extract is preferably a fruit or leaf extract.

It may in particular be a hydroalcoholic, or aqueous, or subcritical CO ₂ extract, or a subcritical H ₂ O extract, or an extract obtained by extraction in combination with heat treatment carried out by standard heating, or at microwave frequency, or under the influence of ultrasound.

The plant/extract ratio is advantageously from 1/1 to 10,000/1, in particular from 1/1 to 200/1.

The extract preferably contains at least 1 wt.% piperine relative to the total weight of the extract.

Thus, the mixture of the composition according to the invention advantageously contains:

- at least piperine and

- at least one molecule selected from apigenin-7-O-glucuronide, chrysanthellin A, chrysanthellin B, caffeic acid, luteolin, maritimethine, eriodictyol, isoocanin, apigenin, luteolin-7-O-glucoside, maritimein, marein, eriodictyol- 7-O-glucoside, flavomarein, apigenin-8-C-α-L-arabinoside-6-C-β-D-glucoside (shaftoside), apigenin-6,8-C-di-β-D-glucopyranoside (vicenin -2) or analogues thereof, preferably apigenin-7-O-glucuronide, and

- at least one molecule selected from dicaffeoylquinic acid, sulfomonocaffeoylquinic acid, luteolin, luteolin-7-O-glucoside, luteolin-7-O-glucuronide, apigenin-7-O-glucoside, cinaropicrin or analogues thereof, preferably dicaffeoylquinic acid, And

- at least one molecule selected from monocaffeylquinic acid, delphinidin-3-galactoside, delphinidin-3-glucoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, cyanidin-3-glucoside, petunidin-3-galactoside, cyanidin- 3-arabinoside, petunidin-3-glucoside, peonidin-3-galactoside, petunidin-3-arabinoside, peonidin-3-glucoside, malvidin-3-galactoside, malvidin-3-glucoside, malvidin-3-arabinoside or analogues thereof, preferably at least one monocaffeylquinic acid.

In addition to the extracts of Chrysanthellum indicum, Cynara scolymus, Vaccinium myrtillus and piperine, the mixture according to the invention may also contain other compounds, in particular an extract of Olea europaea.

The Olea europaea extract is preferably a fruit or leaf extract.

It may in particular be a hydroalcoholic, or aqueous, or subcritical CO ₂ extract, or a subcritical H ₂ O extract, or an extract obtained by extraction in combination with heat treatment carried out by standard heating, or at microwave frequency, or under the influence of ultrasound.

The plant/extract ratio is advantageously from 1/1 to 200/1, in particular from 1/1 to 60/1.

The composition according to the invention intended for humans preferably contains an amount of Olea europaea extract that allows the administration of at least 0.00001 g, in particular from 0.00001 g to 0.60 g, of Olea europaea extract per kg body weight of the person to whom the composition is administered per day.

Preferably, the Olea europaea extract contains at least one molecule selected from oleuropein and hydroxytyrosol or analogues thereof.

For purposes of the invention, the term "mixture" means a combination of substances (extracts and/or molecules) in solid, liquid or gaseous form, which may or may not react chemically.

The extract mixture according to the invention is prepared by any method known to those skilled in the art. It can be obtained by simply mixing the constituent parts.

Preferably, the ratio of Chrysanthellum indicum extract/Cynara scolymus extract/Vaccinium myrtillus extract/piperine in the mixture is from 0.01/0.01/0.01/0.0001 to 10/10/10/10.

According to one embodiment, in addition to or instead of a mixture consisting of several plant extracts, the mixture of molecules of the composition according to the invention may contain at least:

- a single extract obtained from a mixture of at least two plants selected from Chrysanthellum indicum, Cynara scolymus and Vaccinium myrtillus and Piper, and optionally

- Chrysanthellum indicum extract, if the mixture of plants of a single extract does not contain Chrysanthellum indicum,

- Cynara scolymus extract, if the plant mixture of a single extract does not contain Cynara scolymus,

- Vaccinium myrtillus extract, if the mixture of plants of a single extract does not contain Vaccinium myrtillus,

- piperine or Piper extract, if the plant mixture of a single extract does not contain Piper.

Advantageously, when the composition contains a single extract, it contains:

- a single extract obtained from at least Chrysanthellum indicum, Cynara scolymus and Vaccinium myrtillus and Piper, and/or

- piperine and a single extract obtained from at least Chrysanthellum indicum, Cynara scolymus and Vaccinium myrtillus.

For the purposes of the invention, the term “single extract obtained from more than one plant “X” or plant material “X”” means a group of molecules obtained from a mixture of at least two plants “X” by any suitable method. In particular, mention may be made of aqueous extracts (obtained using an aqueous solvent), alcoholic extracts (obtained using an alcohol solvent), either obtained using an organic solvent or using a natural fatty substance or mixture of natural fatty substances, especially vegetable oil or mixture of vegetable oils . The term "aqueous solvent" means any solvent consisting wholly or partially of water. Thus, mention may be made of water itself, hydroalcoholic solvents in any proportions, or solvents consisting of water and a compound such as glycerin or propylene glycol in any proportions. Among alcohol solvents, ethanol may be particularly mentioned.

For the purposes of the invention, the term “plant or plant material” means the whole plant or part of a plant, including cell cultures that have not yet undergone special processing, and intended for inclusion in the production of a herbal preparation.

The single extract of plant mixture "X" may be obtained by any suitable method, for example, by a method comprising the following steps:

- solid-liquid extraction

- separation/pressing

- filtering

- evaporation

- drying

- optional addition of additives

- homogenization

- air conditioning.

Preferably, the whole plant or aerial parts of Chrysanthellum indicum are used as plant raw materials to obtain a single extract. The single extract is preferably prepared from at least 0.1% by weight of the whole plant or aerial parts of Chrysanthellum indicum, relative to the total weight of the plant mixture used to obtain the single extract.

Preferably, the whole plant or aerial parts of Cynara scolymus are used as plant raw materials to obtain a single extract. The single extract is preferably prepared from at least 0.1% by weight of the whole plant or aerial parts of Cynara scolymus, relative to the total weight of the plant mixture used to prepare the single extract.

Preferably, fruits or leaves of Vaccinium myrtillus are used as plant raw materials to obtain a single extract. The single extract is preferably prepared from at least 0.1% by weight of the fruits or leaves of Vaccinium myrtillus, relative to the total weight of the plant mixture used to obtain the single extract.

If a single extract is obtained from a mixture of plants containing Piper, the fruits or leaves of Piper nigrum, Piper aduncum and/or Piper longum are preferably used as plant raw materials to obtain the single extract. The single extract is preferably prepared from at least 0.0001% by weight of the fruits or leaves of Piper nigrum and/or Piper aduncum and/or Piper longum, relative to the total weight of the plant mixture used to obtain the single extract.

The single extract may thus contain, in addition to other molecules, at least oleuropein, hydroxytyrosol and/or at least piperine.

Preferably, the single extract contains:

- at least one molecule selected from apigenin-7-O-glucuronide, chrysanthellin A, chrysanthellin B, caffeic acid, luteolin, maritimethine, eriodictyol, isoocanin, apigenin, luteolin-7-O-glucoside, maritimein, marein, eriodictyol- 7-O-glucoside, flavomarein, apigenin-8-C-α-L-arabinoside-6-C-β-D-glucoside (shaftoside), apigenin-6,8-C-di-β-D-glucopyranoside (vicenin -2) or analogues thereof, preferably apigenin-7-O-glucuronide, and

- at least one molecule selected from dicaffeoylquinic acid, sulfomonocaffeoylquinic acid, luteolin, luteolin-7-O-glucoside, luteolin-7-O-glucuronide, apigenin-7-O-glucoside, cinaropicrin or analogues thereof, preferably diopheoylquinic acid, And

- at least one molecule selected from monocaffeylquinic acid, delphinidin-3-galactoside, delphinidin-3-glucoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, cyanidin-3-glucoside, petunidin-3-galactoside, cyanidin- 3-arabinoside, petunidin-3-glucoside, peonidin-3-galactoside, petunidin-3-arabinoside, peonidin-3-glucoside, malvidin-3-galactoside, malvidin-3-glucoside, malvidin-3-arabinoside or analogues thereof, preferably at least one monocaffeylquinic acid.

- piperine.

In addition to Chrysanthellum indicum, Cynara scolymus and Vaccinium myrtillus, the single extract of the composition according to the invention can also advantageously be obtained from Olea europaea. Preferably, leaves or fruits of Olea europaea are used as plant raw materials to obtain a single extract. The single extract is advantageously prepared from at least 0.1% by weight of the leaves or fruits of Olea europaea, relative to the total weight of the plant mixture used to obtain the single extract.

A particularly suitable embodiment of the invention is a composition containing a single extract obtained from Chrysanthellum indicum, Cynara scolymus, Vaccinium myrtillus, Piper and Olea europaea.

A composition according to the invention containing a single extract intended for humans preferably contains an amount of single extract that corresponds to the administration of at least 0.0001 g, in particular from 0.00001 g to 60 g, of single extract per kg of body weight of the person to whom the composition is administered per day.

The composition according to the invention containing plant extracts and/or one or more unitary extracts thus contains:

- at least one molecule selected from apigenin-7-O-glucuronide, chrysanthellin A, chrysanthellin B, caffeic acid, luteolin, maritimethine, eriodictyol, isoocanin, apigenin, luteolin-7-O-glucoside, maritimein, marein, eriodictyol- 7-O-glucoside, flavomarin, apigenin-8-C-α-L-arabinoside-6-C-β-D-glucoside (shaftoside), apigenin-6,8-C-di-β-D-glucopyranoside (vicenin -2) or analogues thereof, preferably apigenin-7-O-glucuronide, and

- from dicaffeoylquinic acid, sulfomonocaffeoylquinic acid, luteolin, luteolin-7-O-glucoside, luteolin-7-O-glucuronide, apigenin-7-O-glucoside, cinaropicrin or their analogues, preferably dicaffeoylquinic acid, and

- at least one molecule selected from monocaffeylquinic acid, delphinidin-3-galactoside, delphinidin-3-glucoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, cyanidin-3-glucoside, petunidin-3-galactoside, cyanidin- 3-arabinoside, petunidin-3-glucoside, peonidin-3-galactoside, petunidin-3-arabinoside, peonidin-3-glucoside, malvidin-3-galactoside, malvidin-3-glucoside, malvidin-3-arabinoside or analogues thereof, preferably at least one monocaffeylquinic acid, and

- at least piperine,

- and optionally preferably at least one molecule selected from oleuropein, hydroxytyrosol or analogues thereof.

According to another aspect, the invention also relates to a composition containing a mixture of at least four molecules, where these molecules are possibly synthetic molecules and/or natural molecules, especially molecules isolated from plant materials:

- at least one molecule selected from apigenin-7-O-glucuronide, chrysanthellin A, chrysanthellin B, caffeic acid, luteolin, maritimethine, eriodictyol, isoocanin, apigenin, luteolin-7-O-glucoside, maritimein, marein, eriodictyol- 7-O-glucoside, flavomarein, apigenin-8-C-α-L-arabinoside-6-C-β-D-glucoside (shaftoside), apigenin-6,8-C-di-β-D-glucopyranoside (vicenin -2) or their analogues, and

- at least one molecule selected from dicaffeoylquinic acid, sulfomonocaffeoylquinic acid, luteolin, luteolin-7-O-glucoside, luteolin-7-O-glucuronide, apigenin-7-O-glucoside, cinaropicrin or analogues thereof, and

- at least one molecule selected from monocaffeylquinic acid, delphinidin-3-galactoside, delphinidin-3-glucoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, cyanidin-3-glucoside, petunidin-3-galactoside, cyanidin- 3-arabinoside, petunidin-3-glucoside, peonidin-3-galactoside, petunidin-3-arabinoside, peonidin-3-glucoside, malvidin-3-galactoside, malvidin-3-glucoside, malvidin-3-arabinoside or analogues thereof, and

- at least piperine.

At least one of the first three molecules is a synthetic molecule. Indeed, regardless of piperine, which may be synthetic or natural, at least one of the other molecules is a synthetic molecule.

In addition to these at least four molecules, the mixture may also contain at least one molecule selected from oleuropein, hydroxytyrosol or analogues thereof, where these molecules are possibly synthetic or natural molecules.

In a particularly suitable embodiment, the mixture contains at least one dicaffeylquinic acid, apigenin-7-O-glucuronide, monocaffeylquinic acid, piperine and oleuropein. The mixture of molecules of the composition according to the invention may consist solely of dicapheylquinic acid, apigenin-7-O-glucuronide, monocaffeoylquinic acid, piperine and oleuropein.

In other embodiments, the compositions of the invention may contain a mixture of the extract(s) and molecule(s) of interest provided herein (e.g., Chrysanthellum indicum extract and at least one molecule selected from dicaffeoylquinic acid, sulfomonocaffeoylquinic acid, luteolin, luteolin-7-O-glucoside, luteolin-7-O-glucuronide, apigenin-7-O-glucoside, cinaropicrin or analogs thereof, and at least one molecule selected from monocaffeylquinic acid, delphinidin-3-galactoside, delphinidin-3 -glucoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, cyanidin-3-glucoside, petunidin-3-galactoside, cyanidin-3-arabinoside, petunidin-3-glucoside, peonidin-3-galactoside, petunidin-3-arabinoside , peonidin-3-glucoside, malvidin-3-galactoside, malvidin-3-glucoside, malvidin-3-arabinoside or analogues thereof, and at least piperine, etc.).

The compositions according to the invention in their various embodiments may consist exclusively of the described elements (plant extracts and/or single extract(s)) and/or a mixture of at least four molecules) or may also contain at least one additional element (products, molecules, extracts, active substances, excipients, etc.) added in addition to plant extracts and/or single extract(s) or a mixture of at least four molecules, where said additional element possibly selected from:

- the following vitamins: B1, B2, B3, B5, B6, B8, B9, B12, C, A, D, E, K1 and K2;

- the following compounds: obeticholic acid, corosolic acid, polyunsaturated fatty acids of the omega-6 and/or omega-3 family, orotic acid, pangamic acid, para-aminobenzoic acid, amygdalin, beta-glucans, carnitine, dimethylglycine, imeglimin, isoflavones, L -arginine, oxytocin, pectin, pyridoxamine, resveratrol, viniferine, L-citrulline;

- the following trace elements and minerals: arsenic, boron, calcium, copper, iron, fluorine, iodine, lithium, manganese, magnesium, molybdenum, nickel, phosphorus, selenium, vanadium, zinc;

- the following microcomponents of replaceable nature: conjugated linolenic acid, lipoic acid, carotenoids, carnitine, choline, coenzyme Q10, phytosterols, polyphenols of the tannin and lignan family, taurine;

- fructooligosaccharides, galactooligosaccharides;

- bacteria that ferment lactic acid;

- yeast, for example red rice yeast (Monascus purpureus);

- mushrooms, for example curly grifola;

- products isolated from insects that are compatible with the food and pharmaceutical sectors;

- coating agents: for example hypromellose, microcrystalline cellulose, stearic acid, talc, sucrose, shellac, povidone, beeswax;

- flavoring additives: for example natural blueberry flavoring additive or natural strawberry flavoring additive;

- acidulants such as malic acid;

- substances that prevent agglomeration: for example, silicon dioxide or magnesium stearate;

- thickeners such as xanthan gum, colloidal silicon dioxide, mono- and diglycerides of fatty acids;

- stabilizers such as calcium phosphate;

- emulsifiers such as soy lecithin;

- fillers such as corn starch;

- excipients, for example microcrystalline cellulose, magnesium stearate or dicalcium phosphate.

The compositions of the invention may also contain one or more extracts of at least one of the following plant materials, and/or one or more molecules contained in at least one of the following plant materials, and/or a single extract may also be obtained from at least one of the following types of plant materials: Abelmoschus esculentus, Abies alba, Abies balsamea, Abies sibirica, Acacia nilotica, Acacia senegal, Achillea millefollium, Achyranthes bidentata, Acmella oleracea, Actaea racemosa, Actinidia chinensis, Actinidia deliciosa, Adansonia digitata, Adiantum capillus-veneris, Aesculus hippocastanum, Afromomum melegueta, Agathosma betulina, Agathosma crenulata, Agathosma serratifolia, Agrimonia eupatoria, Ajuga reptans, Albizia julibrissin, Alchemilla vulgaris, Alliara petiolata, Allium ampeloprasum, Allium cepa, Allium sativum, Allium scho enoprasum, Allium ursinum, Alnus glutinosa, Aloe ferox, Aloe vera, Aloysia citriodora, Alpinia galanga, Alpinia hainanensis, Alpinia officinarum, Alpinia oxyphylla, Althaea officinalis, Ammi visnaga, Amorphophallus konjac, Ananas comosus, Andographis paniculata, Anemarrhena asphodeloides, Anethum graveolens, Angelica archangelica, Angelica dahurica Angelica pubescens, Angelica sinensis, Antennaria diocia, Anthriscus cerefolium, Anthyllis vulneraria, Aphanizomenon flos-aquae Ralfs, Apium graveolens, Arachis hypogaea, Aralia elata, Arctium lappa, Arctium minus, Argania spinosa, Armorica rustanica, Artemisia dracunculus, Artemesia vulgaris, Ascophyllum nodo sum , Aspalathus linearis, Asparagus officinalis, Astragalus membranaceus, Atractylodes lancea, Atractylodes macrocephala, Auracaria columnaris, Avena sativa, Ayahuasca, Baccharis genistelloides, Bacopa monnierri, Ballota nigra, Bambusa bambos, Bellis perennis, Berberis vulgaris, Beta vulgaris, Betula alleghaniensis, Betula pendula , Betula pubescens, Bixa orellana, Borago officnalis, Boswellia serrata, Brassica napus, Brassica nigra, Brassica oleracea, Brassica rapa, Bupleurum chinense, Calendula officinalis, Calluna vulgaris, Camellia sinsensis, Capsella bursa-pastoris, Capsicum annuum, Carex arenaria, Carica p apaya , Carlina acaulis, Carphephorus odoratissmus, Carpinus betulus, Carthamus tinctorius, Carum carvi, Cassia fistula, Castanea sativa, Centaurea centaurium, Centaurea cyanus, Centaurium erythraea, Centella asiatica, Cerasus vulgaris, Ceratonia silliqua, Chaenomelum nobile, Chlorella vulgaris, Chondrus crispus, Chrysanthellum indicum, Cichorium intybus, Cinchona officinalis, cinchona pubescens, Cinnamomum camphora, Cinnamomum cassia, Cinnamomum verum, Cistanche salsa, Cistus incanus, Citrus aurantium, Citrus limon, Citrus maxima, Citrus medica, Citrus myrtifolia, Citrus reticulata blanco, Citrus sinsensis, Citrus paradisi , Clinopodium vulgare, Cnicus benedictus, Cochlearia officinalis, Cocos nucifera, Codonopsis pilosula, Coffea canephora, Coix lacryma-jobi var. mayyuen Stapf, Cola acuminata, Cola ballayi cornu, Cola nitida, Combretum micranthum, Commiphora mukul, Conyza canadensis, Coriandrum sativum, Cornus officinalis, Corylus avellana, Corymbia citriodora, Crataegus laevigata, Craetegus monogyna, Crithmum maritimum, Crocus sativus, Cucumis me lo, Cucurbita pepo, Cuminum cyminum, Cupressus sempervirens, Cuscuta chinensis, Cyamopsis tetragonoloba, Cyathula officinalis, Cyclanthera pedata, Cydonia oblonga, Cymbopogon martini, Cymbopogon nardus, Cymbopogon winterianus, Cynara cardunculus, Cyperus rotundus, Daucus carota, Dendranthema grandiflorum, Desmodium adscendens, Dimocarpus longan, Dioscorea oppostifolia, Dioscorea villosa, Diospyros kaki Thunb., Dunaliella saliena, Echinacea augustifolia, Echinacea pallida, Echinacea purpurea, Elaegnus rhamnoides, Alettaria cardamomum, Eleutherococcus senticosus, Elymus repens, Epiobium augustifolium, Epilobium parviflorum, Equisetum arvense, Erica cinerea, Erica tetralix, Eriobotrya japonica, Eriodictyon californicum, Erodium cicutarium, Eryngium campestre, Eschscholzia californica, Eucalyptus dives Schauer, Eucalyptus globulus, Eucalyptus radiata, Eucalyptus smithii F. Muell, Eucommia ulmoides, Eugenia uniflora, Eugenia jambolana, Euphrasia stricta D. Wolff, Euterpe oleracea, Fagopyrum esculentum Moench, Follopia japonica, Ferula assa-foetida, Ficus carica, Filipendula ulmaria, Foeniculum vulgare Mill., Forsythia suspensa, Fragaria dodonei Ard., Frangula purshiana Cooper, Fraxinus excelsior, Fraxinus ortus, Fucus serratus, Fucus vesiculosus, Fumaria officinalis, Galeo psis segetum Neck., Galium odotarum, Galium verum, Gardenia jasminoides J. Ellis, Gastrodia elata Blume, Gelidium corneum J.V. Lamouroux, Gentiana lutea, Geranium robertianum, Geum urbanum, Ginkgo biloba, Glycine max, Glycyrrhiza glabra, Glycyrrhiza uralensis, Gracilaria gracilis, Grindelia camporum Greene, Grindelia robusta Nutt., Grindelia squarrosa Dunal, Gymnema sylvestris, Haematococcus pluvialis, Hamamemis virginiana, Harpagophytum procumbens , Harpagophytum zeyheri Decne., Hedeoma pluegioides Pers., Helianthus annuus, Helienthus tuberosus, Helichrysum arenarium, Helichrysum stoechas, Herniara glabra, Hibiscus sabdariffa, Hieracium pilosella, Himanthalia elongata, Hordeum vulgare, Houttuynia cordata Thunb. , Huperzia serrata, Hyssopus officinalis, Ilex paraguariensis A. St.-Hill, Illicum verum, Impatients balsamina, Inula britannica, Inula helenium, Jasminum grandiflorum, Jasmium officinale, Juniperus communis, Justicia adhatoda, Kavalama urens, Krameria lappacea, Lagerstroemia speciosa, Laminaria digitata, Laminaria hyperborea, Lamium album, Larix decidua, Larix occidentalis, Laurus nobilis, Lavandula augustofolia, Lavandula latifolia, Ledum palustre, Leonurus cardiaca, Lepidium meyenii Walp., Lepidium sativum, Lespedeza capitata, Levisticum officinale, Lindera aggregata, Linus usitatissimum, Liquidambar styraciflua, Lotus corniculatus, Lycium chinense, Lycium barbarum, Lycopersicon esculentum, Lycopodium clavatum, Lycopus europaeus, Lythrum salicaria, Macadamia ternifolia F. muell, Macrocystis pyrifera, Magnolia officinalis, Malpighia glabra, Malus pumila, Malus domestica, Malus sylvestris, Malva sylvestris, Mangifera indica, Maranta arun dinacea, Marrubium vulgare , Marsdenia cundurango, Marsdenia sylvestris, Mastocarpus stellatus, Matricaria chamomilla, Medicago sativa, Melaleuca alternifolia, Melaleuca cajuputi Powell, Melaleuca leucadendra, Melaleuca quinquenrvia, Melaleuca viridiflora, Melilotus altissimus Thuill., Melilotus officinalis, Mentha arven sis, Mentha x piperita, Menyanthes trifoliata, Mesembryanthemum crystallinum, Monarda didyma, Morinda citrifolia, Morinda officinalis, Morus alba, Morus nigra, Murraya koenigii, Musa x paradisiaca, Myrciaria dubia, Myristica flagrans Houtt., Myroxylon balsamum, Myrtus communis, Nardostachys jatamansi, Nasturtium officinale R. Br., Nelumbo nucifera Gaertn., Nepeta cataria, Nepeta tenuifolia Benth., Nigella sativa, Ocimum basilicum, Oenothera biennis, Ononis spinosa, ophiopogon japonicus, Opuntia ficus-indica, Origanum compactum Benth., Origanum majorana, Origanum vulgare, Orthosiphon aristatus, Oryza sativa, Paeonia lactiflora, Paeonia x suffruticosa Andrews, Palmaria palmata, Panax ginseng, Panax quinquefolius, Panicum miliacium, Papaver rhoeas, Parietaria officinalis, Passiflora edulis Sims, Pastinaca sativa, Paullinia cupana Kunth, Pelargonium graveolens, Perilla frutescens, Persea americana, Persicaria bistorta , Persicaria maculosa Gray, Petroselinum crispum, Peucadanum ostruthium, Peumus boldus Molina, Phaseolus vulgaris, Phellodendron amurense, Photinia melancarpa, Phyllanthus emblica, Physalis alkekengi, Phymatolithon calcareum, Picea abies, Pimenta dioca, Pimenta racemosa, Pimpinella anisum, Pimpinella major, Pimpinella saxfraga, Pinus mugo Turra, Pinus pinaster Aiton, Pinus sylvestris, Pistacia lentiscus, Plantago arenaria, Plantago lanceolata, Plantago major, Plantago ovata, Platycodon grandiflorus, Plectranthus barbatus Andrews, Pogostemom cablin, Polygala senega, Polygala sibirica, Polygala tenuifolia Willd., Polygonum aviculare, Populus nigra , Populus tremula, Populus tremuloides, Porphyra umbilicalis, Portulaca oleracea, Potentilla erecta, Primula veris, Prunella vulgaris, Prunus africana, Prunus armeniaca, Ribes nigrum, Ribes uva-crispa, Rosa canina, Rosa gallica, Rosa moschata, Rosa rubiginosa, Rosmarinus officinalis , Rubus caesius, Rubus fruticosus, Rubus idaeus, Rumex actetosa, Rumex acetosella, Rumex crispus, Rumex patienta, Ruscus aculeatus, Sachharina japonica, Saccharina latissima, Salix alba, Salix fragilis, Salix pentandra, Salix purpurea, Salvia officinalis L., Salvia officinalis subsp. lavandulifolia Gams, Salvia sclarea, Sambucus nigra, Sanguisorba officinalis, Sanicula elata Buch.-Ham. Ex D. Don, Santalum album, Santolina chamaecyparissus, Saposhnikovia divaricata, Sargassum fusiforme, Satureja hortensis, Satureja montana, Saussurea costus, Scrophularia ningpoensis Helmsl., Scutellaria baicalensis Georgi, Secale cereale, Sedum acre, Sedum roseum, Senna alexandrina Mill., Senna obustifolia, Smilax cordifolia Humb. & Bonpl., Smilax glabra Roxb., Smilax officinalis Kunth, Smilax purhampuy Ruiz, Smilax purhampuy Ruiz, Smilax regelli Killip & C.V. Morton, Smilax vanillidora Apt, Solanum melongena, Solanum tuberosum, Solidago virgaurea, Sorbus aucuparia, Spatholobus suberctus Dunn., Spinacia oleracea, Spirulina major Kützing, Spirulina maxima Geitler, Spirulina platensis Geitler, Stavhys officinalis, Stemmacantha carthamoides Dittrich, Stypholo bium japonicum, Syzgium aromaticum , Tagetes erecta, Tamarindus indica, Tanacetum parthemium, Terminalia chebula Retz., Theobroma cacao, Thymus saturejoides Coss., Thymus serpyllum, Thymus vulgaris, Thymus zygis, Tilia cordata Mill., Tilia platyphyllos Scop., Tilia tomentosa Moench, Tilia euopaea, Tribulus Terrestris, Trichosanthes Kirilowii Maxim., Trifolium arvest, Trifolium Campestre Schreb., Trifolium Pratense, Trifolium Repens, Trigonella Caerulea, Trigonella Foneenum- Graecum, Tricitum aestivum, Tricitum Durum Desf., Tricitum spelta L., Tricitum Turgidum, Tropaeolum Majus, Turnra Diffusa Willd., Ulmus glabra Huds., Ulmus glabra Huds., Ulmus pumila, Ulmus rubra Muhl., Ulva lactuca, Uncaria gambir Roxb., Uncaria rhynchophylla Miq., Uncaria tomentosa DC., Undaria pinnatifida Suringar, Urtica dioca, Urtica urens, Vaccinium macrocarpon, Vaccinium oxycoccos, Vaccinium vitis-idae, Valeriana jatamansi Jones, Valeriana officinalis, Vanilla planifolia Jacks, Verbascum densiflorum Bertol., Verbascum thapsus, Verbena officinalis, Veronica officinalis, Viburnum opulus, Vigna angularis Ohwi & H. Ohashi, Vinca major, Vinca minor, Viola palustris, Viola tricolor, Vitex agnus-castus, Vitex trifolia, Vitis vinifera, Zea mays, Zingiber officinale Roscoe, Ziziphus jujuba Mill.

The compositions according to the invention may be in any form, especially in the form of a powder, gel, emulsion or liquid, in particular in the form of tablets, wafers, gelatin capsules, sticks, sachets, vials, droppers or injectable form.

The compositions according to the invention can be used as food or health products, in particular as medicines.

The term "food product" means a product having a nutritional and/or physiological effect, it particularly includes nutritional supplements, food products, dietary products, etc. These products can in particular be administered orally, intragastrically or intravenously.

The term "health product" means any product that has a beneficial effect on a health condition, either in prevention or treatment, regardless of whether this effect is physiological or pharmacological, especially drugs, pharmaceuticals. These products can in particular be administered orally, intragastrically, intravenously or subcutaneously.

The compositions according to the invention can be used for the prevention and/or control of carbohydrate and/or fat metabolism disorders in humans or animals.

They are particularly suitable for the prevention and/or control of type 2 diabetes mellitus in humans or animals. In particular, they make it possible to prevent the development of chronic hyperglycemia, reduce fasting glycemia and levels of glycated hemoglobin, circulating and hepatic triglycerides, body weight and adipose tissue mass, increase high-density lipoprotein (HDL) cholesterol ("good cholesterol") and increase tolerance to ingested carbohydrates and insulin sensitivity. They can be used prophylactically in the treatment of type 2 diabetes and as first-line therapy while taking HDM, thereby delaying the use of conventional oral antidiabetic molecules. They are also particularly suitable for the treatment of type 2 diabetes mellitus and its complications, especially non-alcoholic steatohepatitis (NASH), as monotherapy or in combination with other pharmacological agents.

The compositions according to the invention can also be used for the prevention and/or control of type 1 diabetes mellitus and/or non-alcoholic fatty liver diseases, in particular non-alcoholic steatohepatitis (NASH), and/or cardiovascular pathologies, in particular coronary cardiopathy, cerebrovascular disease, peripheral arteriopathy, deep vein thrombosis and/or pathologies associated with insulin resistance, such as Alzheimer's disease (Bedse G et al. Front Neurosci 2015; 9:204).

For such applications, the compositions of the invention may be used in combination with at least one antidiabetic therapeutic agent selected from biguanides including metformin, dipeptidyl peptidase-IV (DPP-IV) inhibitors, glucagon-like peptide-1 (GLP-1) analogues, thiazolidinediones (TZDs) ), sulfonylureas, short-acting and long-acting insulins, glucosidase inhibitors (acarbose, miglitol, voglibose), sodium-dependent glucose cotransporter 2 (SGLT2), fibranor family molecules such as elafibranor, or molecules targeting nuclear receptors, and in features on ROR receptors (α, β, γ) and Rev-Erb receptors (α, β).

The compositions of the invention may also be used to target other risk factors for cardiovascular disease or metabolic syndrome.

In particular, the compositions according to the invention can be used for the prevention and/or control of dyslipidemia. They particularly have a cholesterol-lowering effect and can reduce total cholesterol, low-density lipoprotein (LDL) cholesterol, circulating triglycerides and hepatic triglycerides. They also have inhibitory activity against 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA).

For such applications, the compositions of the invention can be used in combination with a lipid-lowering therapeutic agent selected from: statins, fibrates, niacin, ion exchange resins, cholesterol absorption inhibitors, omega-3 polyunsaturated fatty acids, thiadenol and nuclear farnesoid X receptor (FXR) agonists. .

Finally, the compositions according to the invention can be used, in particular, for the prevention or control of obesity and overweight, and/or metabolic syndrome, and/or abnormal blood pressure problems.

The invention will now be illustrated by examples of extracts and compositions, as well as test results demonstrating the effectiveness of the compositions according to the invention, these examples and tests being non-limiting.

I. Examples

Example 1: Chrysanthellum indicum dry extract example

The aerial parts of a fresh or dried plant are mechanically crushed to obtain a coarse powder. This powder is then subjected to a maceration step for 10 to 24 hours at room temperature in a 70/10 water/ethanol mixture, then the resulting mixture is subjected to continuous leaching at 50 °C in a percolator with a 70/10 water/ethanol mixture, the ratio plant/extract is 3/1. The resulting extract is then subjected to liquid-liquid washes using a non-polar organic solvent such as dichloromethane or trichloromethane. After concentration by low pressure evaporation at 35 °C, a liquid is obtained which, after lyophilization for 24 hours, forms a beige powder that is soluble in a water/alcohol mixture. This powder (dry extract) can be applied directly or mixed in a suitable solvent before use.

Example 2: Example of Vaccinium myrtillus dry extract

Blueberries in the form of a powder obtained from the fruits of Vaccinium myrtillus are subjected to a maceration step for 10 to 24 hours at room temperature in a mixture of 30/50 water/ethanol, and then the resulting mixture is subjected to continuous leaching at 50 °C in a percolator with a mixture of 30/ 50 water/ethanol, with a plant/extract ratio of 10/1. The resulting extract is then subjected to liquid-liquid washes using a non-polar organic solvent such as dichloromethane or trichloromethane. After concentration by low pressure evaporation at 35 °C, a liquid is obtained which, after lyophilization for 24 hours, forms a purple powder that is soluble in a water/alcohol mixture.

Example 3: Example of Cynara scolymus dry extract

The artichoke in the form of a powder obtained from the leaves of Cynara scolymus is subjected to a maceration step for 10 to 24 hours at room temperature in water, and then the resulting mixture is subjected to continuous leaching at 50 °C in a percolator with water, the plant/extract ratio being 2/1. The resulting extract is then subjected to liquid-liquid washes using a non-polar organic solvent such as dichloromethane or trichloromethane. After concentration by low pressure evaporation at 35 °C, a liquid is obtained which, after lyophilization for 24 hours, forms a beige powder that is soluble in a water/alcohol mixture.

Example 4: Example of Olea europaea dry extract

Air-dried whole olive leaves are ground at minus 80°C using a knife mill to obtain a fine and homogeneous powder. The resulting powder is then subjected to a maceration step for 10 to 24 hours in a 70/30 water/ethanol mixture. This step is performed in a closed system with nitrogen bubbling at room temperature, or in a microwave reactor at 800 Watts, or at an ultrasonic frequency of 20 kHz for 2 × 3 min. The resulting mixture is then continuously leached at 50°C in a percolator with a 70/30 water/ethanol mixture where the plant/extract ratio is 10/1. The resulting extract is then subjected to liquid-liquid washes using a non-polar organic solvent such as dichloromethane or trichloromethane. After concentration by low pressure evaporation at 35 °C, a liquid is obtained which, after lyophilization for 24 hours, forms a green powder that is soluble in a water/alcohol mixture.

Example 5: Single Extract Examples

Chrysanthemum in the form of powder obtained from the aerial parts of Chrysanthellum indicum, artichoke in the form of powder obtained from the leaves of Cynara scolymus, blueberry in the form of powder obtained from the fruits of Vaccinium myrtillus, powder of the fruits of Piper nigrum and olive tree in the form of powder obtained from the leaves of Olea europaea , are subjected to a maceration step for 10 to 24 hours at room temperature in a 40/60 water/ethanol mixture, and then the resulting mixture is subjected to continuous leaching at 50 °C in a percolator with a 40/60 water/ethanol mixture, where the plant/single ratio the extract ranges from 4 to 6/1. The resulting extract is then subjected to liquid-liquid washes using a non-polar organic solvent such as dichloromethane or trichloromethane. After concentration by low pressure evaporation at 35 °C, a liquid is obtained which, after lyophilization for 24 hours, forms a purple powder that is soluble in a water/alcohol mixture.

Example 6: example of a composition according to the invention in tablet form containing four plant extracts

The composition of Example 6 is in the form of tablets that can be administered orally. It contains, in wt.% relative to the total weight of the composition, 30.1% dry extract of the aerial parts of Chrysanthellum indicum, 30.1% dry extract of the leaves of Cynara scolymus, 3.0% dry extract of the fruits of Vaccinium myrtillus and 0.3% dry extract of the fruits of Piper nigrum . It also contains excipients, in particular microcrystalline cellulose and magnesium stearate.

The composition for three tablets is shown in Table 1 below.

Table 1. Example of a composition in tablet form List of ingredients For three tablets Dry extract of aerial parts of Chrysanthellum indicum 600 mg Dry leaf extract of Cynara scolymus 600 mg Dry fruit extract of Vaccinium myrtillus 60 mg Piper nigrum fruit dry extract 6 mg Microcrystalline cellulose 700 mg Magnesium stearate 26 mg

Example 7: example of a composition according to the invention in the form of gelatin capsules containing five plant extracts

The composition of Example 7 is in the form of gelatin capsules, which can be administered orally. It contains, in wt.% relative to the total weight of the composition, 37.0% dry extract of the aerial parts of Chrysanthellum indicum, 37.0% dry extract of the leaves of Cynara scolymus, 3.7% dry extract of the fruits of Vaccinium myrtillus, 0.004% dry extract of the fruits of Piper nigrum and 22 .2% dry extract of Olea europaea leaves.

The composition for three gelatin capsules is shown in Table 2 below.

Table 2. Example of a composition in the form of a gelatin capsule List of ingredients For three gelatin capsules Dry extract of aerial parts of Chrysanthellum indicum 200 mg Dry leaf extract of Cynara scolymus 200 mg Dry fruit extract of Vaccinium myrtillus 20 mg Piper nigrum fruit dry extract 0.02 mg Olea europaea leaf dry extract 120 mg

Example 8: Example of a composition according to the invention in tablet form containing five plant extracts

The composition of Example 8 is in the form of tablets that can be administered orally. It contains, in wt.% relative to the total weight of the composition, 22.0% dry extract of the aerial parts of Chrysanthellum indicum, 22.0% dry extract of the leaves of Cynara scolymus, 2.2% dry extract of the fruits of Vaccinium myrtillus, 13.2% dry extract of the fruits of Olea europaea and 0.2% dry extract of Piper nigrum leaves. The composition also contains, in addition to the mixture, zinc, vitamins B9, PP, B5, H, B12, D, B6, B2, B2 and chromium. It also contains excipients, in particular dicalcium phosphate, microcrystalline cellulose and magnesium stearate.

The composition for one tablet is shown in Table 3 below.

Table 3. Example of a composition in tablet form List of ingredients In mg per tablet Reference nutritional value for one tablet Dry extract of aerial parts of Chrysanthellum indicum 220.0 _ Dry leaf extract of Cynara scolymus 220.0 _ Dry fruit extract of Vaccinium myrtillus 22.0 _ Olea europaea leaf dry extract 132.0 _ Piper nigrum fruit dry extract 2.0 _ Dicalcium phosphate 198.0 _ Microcrystalline cellulose 153.44 _ Magnesium stearate 20.0 _ Zinc bisglycinate 12.99 33% Vitamin B9 - folic acid 7.30 33% Vitamin PP - nicotinamide 5.30 33.1% Vitamin B5 - calcium pantothenate 2.24 33.3 Vitamin H - biotin 1.66 33.2% Vitamin B12 0.83 33.2% Vitamin D3 0.64 32% Vitamin B6 - pyridoxine hydrochloride 0.56 32.9% Vitamin B1 - thiamine hydrochloride 0.48 32.7% Vitamin B2 - riboflavin 0.45 32.1% Chromium picolinate 0.11 32.5%

Example 9: Example of a composition according to the invention in tablet form containing four plant extracts

The composition of Example 9 is in the form of tablets that can be administered orally. It contains, in wt.% relative to the total weight of the composition, 23.9% dry extract of whole plants of Chrysanthellum indicum, 23.9% dry extract of leaves of Cynara scolymus, 23.9% dry extract of Vaccinium myrtillus fruits and 0.2% dry extract of Piper nigrum fruits . The composition also contains vitamin B1, vitamin B3, pantothenic acid (vitamin B5), zinc and chromium. It contains microcrystalline cellulose and dicalcium phosphate as excipients. It contains zinc, sugar, shellac, povidone and beeswax as coating agents.

The composition for three tablets is shown in Table 4 below.

Table 4. Example of a composition in tablet form List of ingredients For three tablets Reference nutritional values for three tablets Dry fruit extract of Vaccinium myrtillus 600 mg _ Chrysanthellum indicum whole plant dry extract 600 mg _ Dry leaf extract of Cynara scolymus 600 mg _ Piper nigrum fruit dry extract 5 mg _ Vitamin B1 1.1 mg 100 % Vitamin B3 (inositol hexanicotinate) 16 mg (nicotinic acid) 100 % Pantothenic acid (vitamin B5) 6 mg 100 % Zinc (zinc bisglycinate) 5 mg 50 % Chromium (Chromium Picolinate) 40 mcg 100 % Microcrystalline cellulose 600 mg _ Dicalcium phosphate 75 mg _

Example 10: example of a composition according to the invention in the form of a powder for reconstitution in water, conditioned in stick form, containing four plant extracts

The composition of Example 10 is in the form of a powder for reconstitution in water, conditioned in the form of sticks that can be administered orally. It contains, in wt.% relative to the total weight of the composition, 36.3% dry extract of whole plants of Chrysanthellum indicum, 24.2% dry extract of roots of Cynara scolymus, 36.3% dry extract of Vaccinium myrtillus fruits and 2.4% dry extract of Piper longum fruits . The composition also contains vitamin B1, vitamin B3, pantothenic acid (vitamin B5), zinc and chromium. As a flavoring additive, it contains natural blueberry flavoring. It contains malic acid as an acidulant. It contains silicon dioxide as an anti-agglomeration agent, xanthan gum as a thickener and calcium phosphate as a stabilizer.

This composition is listed in Table 5 below.

Table 5. Example of a composition in the form of a powder for dilution in water, conditioned in the form of sticks List of ingredients For three sticks Reference nutritional value for three sticks Dry fruit extract of Vaccinium myrtillus 1500 mg _ Chrysanthellum indicum whole plant dry extract 1500 mg _ Dry root extract of Cynara scolymus 1000 mg _ Piper longum fruit dry extract 100 mg _ Vitamin B1 1.1 mg 100 % Vitamin B3 (inositol hexanicotinate) 16 mg (nicotinic acid) 100 % Pantothenic acid (vitamin B5) 6 mg 100 % Zinc (zinc bisglycinate) 5 mg 50 % Chromium (Chromium Picolinate) 40 mcg 100 %

Example 11: Example of a composition according to the invention in the form of a powder for reconstitution in water, conditioned in the form of sticks

The composition of Example 11 is in the form of a powder for reconstitution in water, conditioned in the form of sticks that can be administered orally. It contains, in wt.% relative to the total weight of the composition, 37.0% dry extract of the aerial parts of Chrysanthellum indicum, 37.0% dry extract of the leaves of Cynara scolymus, 3.7% dry extract of the fruits of Vaccinium myrtillus, 0.2% dry extract of the fruits of Piper nigrum and 22.2% dry extract of Olea europaea leaves. The composition also contains vitamin B12 and chromium. As a flavoring additive, it contains natural strawberry flavoring. It contains silicon dioxide as an anti-agglomeration agent. This composition does not contain any anti-agglomeration agent or thickener.

The composition of such a product is shown in Table 6 below.

Table 6. Example of a composition in the form of a powder for dilution in water, conditioned in the form of sticks List of ingredients For three sticks Reference nutritional value for three sticks Dry fruit extract of Vaccinium myrtillus 120 mg _ Dry extract of aerial parts of Chrysanthellum indicum 1200 mg _ Dry leaf extract of Cynara scolymus 1200 mg _ Piper nigrum fruit dry extract 6 mg Olea europaea leaf dry extract 720 mg _ Vitamin B3 (inositol hexanicotinate) 2.5 mcg 100 % Chromium (Chromium Picolinate) 20 mcg 50 %

Example 12: Example of a composition according to the invention in the form of gelatin capsules

The composition of Example 12 is in the form of gelatin capsules, which can be administered orally. It contains, in wt.% relative to the total weight of the composition, 31.6% dry extract of the aerial parts of Chrysanthellum indicum, 47.4% dry extract of the leaves of Cynara scolymus, 15.8% dry extract of the fruits of Vaccinium myrtillus and 2.6% piperine. The composition also contains vitamin B3 and zinc. It contains soy lecithin derived from non-GMO products as an emulsifier, colloidal silicon dioxide and mono- and diglycerides of fatty acids as thickeners. The capsule is fish gelatin with glycerin and red iron oxide dye.

The composition of such a product is shown in Table 7 below.

Table 7. Example of a composition in the form of a gelatin capsule List of ingredients For two gel capsules Reference nutrition facts for two gelatin capsules Dry fruit extract of Vaccinium myrtillus 100 mg _ Dry extract of aerial parts of Chrysanthellum indicum 200 mg _ Dry root extract of Cynara scolymus 300 mg _ Piperine 15 mg _ Vitamin B3 8 mg (nicotinic acid) 50 % Zinc (zinc gluconate) 10 mg 100 %

Example 13: Example of a composition according to the invention in the form of gelatin capsules

The composition of Example 13 is in the form of gelatin capsules, which can be administered orally. It contains, in wt.% relative to the total weight of the composition, 10.4% dry extract of whole plants of Chrysanthellum indicum, 20.7% dry extract of Cynara scolymus roots, 62.1% dry extract of Vaccinium myrtillus fruits and 2.6% piperine. It contains corn starch as a filler. It contains silicon dioxide and magnesium stearate as an anti-agglomeration agent. The gelatin capsule is of plant origin.

The composition of such a product is shown in Table 8 below.

Table 8. Example of a composition in the form of a gelatin capsule List of ingredients For one gelatin capsule Reference nutritional value for one gelatin capsule Dry fruit extract of Vaccinium myrtillus 300 mg _ Chrysanthellum indicum whole plant dry extract 50 mg _ Dry leaf extract of Cynara scolymus 100 mg _ Piperine 15 mg _

Example 14: example of a composition according to the invention in the form of gelatin capsules containing a single extract

The composition of Example 14 is in the form of gelatin capsules, which can be administered orally. It contains a single hydroalcoholic extract of a powder mixture obtained from the aerial parts of Chrysanthellum indicum, from the leaves of Cynara scolymus, from the fruits of Vaccinium myrtillus, from the fruits of Piper nigrum and from the leaves of Olea europaea. The ratio between the three plants is 1/1/0.1/0.0001/0.6.

A single extract of a mixture of plants is obtained by a method including the following steps:

- solid-liquid extraction

- separation/pressing

- filtering

- evaporation

- drying

- optional addition of additives

- homogenization

-conditioning.

Example 15: example of a composition according to the invention in the form of tablets containing synthetic or plant-derived molecules

The composition of Example 15 is in the form of film-coated tablets that can be administered orally. It contains as active substance per tablet: 50 mg of apigenin-7-O-glucuronide, 50 mg of dicaffeylquinic acid, 100 mg of monocaffeylquinic acid, 10 mg of piperine and 250 mg of oleuropein. Other components used as excipients are: pregelatinized starch, sodium carboxymethyl starch (type A), stearic acid, povidone K90, anhydrous colloidal silicon dioxide.

The active substances may be synthetic, either derived from plant materials or obtained from plant extracts by purification by high performance liquid chromatography.

I. Evaluation of the effectiveness of the composition in vivo

To demonstrate the effect of the compositions according to the invention, in particular on fasting glycemia, glycated hemoglobin, carbohydrate tolerance, insulin sensitivity, body weight and adipose tissue mass and on circulating and liver fats, in vivo experiments were carried out in mice. Molecular biological assessments were also performed. Finally, the compositions were compared with reference pharmacological drugs already on the market or in development.

Experiments were performed on db/db mice. db/db mice have a mutation in leptin receptors that causes disruption of cellular signal transmission of the latter. Leptin receptors undergo high expression in the hypothalamus. Mice with mutations in these receptors are unable to effectively regulate energy storage. This results in high insulinemia from the first days of life (10-14 days) and obesity from 3 to 4 weeks with increased glycemia. These mice are characterized by insulin resistance, hypertriglyceridemia, and glucose intolerance. They constitute a well-fitted and predictive model, especially for the study of insulin sensitivity, triglyceridemia, type 2 diabetes mellitus and one of its complications, non-alcoholic steatohepatitis (NASH) (Aileen JF King Br J Pharmacol 2012; 166(3):877- 894; Sanches SC et al. Biomed Res Int 2015).

II.1 Tests A

The duration of the experiment was nine weeks with an introductory period of one week followed by eight weeks of supplementation with plant extracts and composition X. At the beginning of the experiment, the male mice were ten weeks old.

Nine formulations of X were tested. These formulations were directly incorporated into rodent food, allowing for "multi-purpose" efficacy and wide-scale use of the composition since intravenous or intraperitoneal injections are limited to the small number of people administering this route of administration. This also avoided daily force feeding, which alters various physiological processes.

The compositions tested were as follows:

C1: Chrysanthellum indicum (whole plant) plus piperine (1% and 0.1% of feed, respectively);

C2: Cynara scolymus (leaves) plus piperine (1% and 0.1% of feed, respectively);

C3: Vaccinium myrtillus (fruit) plus piperine (1% and 0.1% of feed, respectively);

C4: Chrysanthellum indicum (whole plant) plus Cynara scolymus (leaves) plus Vaccinium myrtillus (fruit) plus piperine (1%, 1%, 1% and 0.1% feed, respectively);

C5: piperine (0.1% of feed respectively);

C6: Chrysanthellum indicum (whole plant) plus Cynara scolymus (leaves) plus Vaccinium myrtillus (fruit) plus piperine (1%, 1% and 1% of feed respectively);

C7: Chrysanthellum indicum (whole plant) plus Cynara scolymus (leaves) plus Vaccinium myrtillus (fruit) plus piperine (1%, 1%, 1% and 0.001% feed, respectively);

C8: Olea europaea (leaves) (0.6% of feed respectively);

C9: Chrysanthellum indicum (whole plant) plus Cynara scolymus (leaves) plus Vaccinium myrtillus (fruit) plus piperine plus Olea europaea (leaves) (1%, 1%, 1%, 0.001%, 0.6% feed respectively);

The term "piperine" refers to either synthetic piperine or a standardized Piper extract containing 95% piperine. As for other plants, they are dry extracts obtained from plant materials.

The experiments were carried out in several stages. As a result, several “control without restrictions” groups were created. The results obtained for these groups were combined. Compositions C1, C2, C3, C4, C5, C8 and C9 caused similar reductions in food intake. As a result, a “matched control” group was created, i.e. a group consuming the same amount of food per day as groups C1, C2, C3, C4, C5, C8 and C9, so that the results of equivalent consumption could be compared food.

Experimental evaluations have focused in particular on:

Measuring body weight;

Measuring fasting blood glucose;

Changes in glycemia during an oral carbohydrate tolerance test. After force-feeding starch (3 g/kg) in the fasting state, changes in glycemia in response to starch were measured in the tail by biopsy immediately before force-feeding (t0) and then after 30, 60, 90 and 120 minutes. The area under the curve (AUC) was calculated. An increase in AUC reflects carbohydrate intolerance, a decrease reflects an improvement in carbohydrate tolerance.

For compositions C7-C9, an insulin sensitivity test was also performed. The test consisted of an intraperitoneal injection of insulin (2 units/kg) in a fasting state. The change in glycemia in response to insulin injection was measured in the tail by biopsy immediately before injection (t0) and then after 30, 60, 90 and 120 minutes. The area under the curve (AUC) was calculated. A decrease in AUC reflects a better response to insulin injection and thus an improvement in insulin sensitivity. In contrast, an increase in AUC reflects worse insulin sensitivity and therefore insulin resistance.

The assessments presented were conducted immediately before supplementation (t = 0) and at the end of supplementation (t = 8 weeks).

The results obtained are presented in the tables:

Table 9 - effect on insulin sensitivity after eight weeks of administration,

Table 10 - effect on fasting blood glucose after eight weeks of administration,

Table 11 - effect on carbohydrate tolerance after eight weeks of administration,

Table 12 - Effect on body weight after eight weeks of administration.

Insulin sensitivity

Table 9. Effect of the compositions on insulin sensitivity after eight weeks of administration Compositions AUC (week 8 - week 0, in mg×min/dl)
(mean ± standard error of the mean (SEM)) control without restrictions 19 186 ± 3460 C7 2082±2082 ^** C8 8970 ± 2959 C9 -6520±4685 ^**

Mean values ± SEM. Unlimited control, n equals 20; C7, n is 11; C8, n equals 10; C9, n equals 9. Composition versus control, Student's t test for unpaired data, ^** p less than 0.01.

The results presented in Table 9 are also illustrated in FIG. 1. These results show a highly statistically significant and potent effect of compositions C7 and C9 on insulin sensitivity. The improvement of insulin sensitivity by compositions C7 and C9 according to the invention is of particular benefit in the prevention and treatment of type 2 diabetes mellitus and its complications. In particular, insulin resistance is one of the main mechanisms of progression of type 2 diabetes mellitus (Samuel VT et al. Cell 2012; 148:852-71). In addition, by influencing this parameter, the compositions C7 and C9 according to the invention are particularly suitable for other pathologies in which insulin resistance is one of the main causes. In particular, this applies to one of the complications of type 2 diabetes, non-alcoholic steatohepatitis (NASH; Samuel VT et al. Cell 2012; 148:852-71) and Alzheimer's disease (Bedse G et al. Front Neurosci 2015; 9: 204).

Fasting glycemia

Table 10. Effect of the composition on fasting glycemia after eight weeks of administration Compositions Fasting blood glucose (in mg/dL) control without restrictions 534 ± 10 control of equal feeding 518.2 ± 38 C1 410.4 ± 33 ^* C2 448 ± 34 C3 384 ± 40 ^* C4 335±38 ^** C5 449 ± 11 C6 536 ± 15 C7 571 ± 14 ^* C8 400 ± 61 C9 220±78 ^**

Mean values ± SEM. control without restrictions, n equals 32; equal feeding control, n equals 10; C1, n is 9; C2, n is 9; C3, n is 9; C4, n is 8; C5, n is 9; C6, n equals 14; C7, n is 12; C8, n is 12; C9, n equals 9. Composition versus control, Student's t test for unpaired data, ^* p less than 0.05, ^** p less than 0.01.

Compositions C1, C2, C3, C4, C5, C8 and C9 caused a decrease in food intake. As a result, a group was created that consumed the same amount of food. Results show that calorie restriction had no effect on fasting glycemia (comparing unrestricted versus same-fed controls). Compositions C1, C3, C4 and C9 caused a statistically significant decrease in fasting glycemia, with the most pronounced effect observed with the introduction of composition C4 (1% Chrysanthellum indicum, 1% Cynara scolymus, 1% Vaccinium myrtillus, 0.1% piperine) and C9 ( 1% Chrysanthellum indicum, 1% Cynara scolymus, 1% Vaccinium myrtillus, 0.001% piperine, 0.6% Olea europaea).

Composition C6 (1% Chrysanthellum indicum, 1% Cynara scolymus, 1% Vaccinium myrtillus) had no effect on fasting blood glucose. Adding piperine (0.1%), although it (C5) did not produce a statistically significant reduction in fasting blood glucose, to the combination of C6 to form combination C4 unexpectedly resulted in a statistically significant and robust reduction in fasting blood glucose. Similarly, adding Olea europaea (0.6%; C8) to the C7 combination (1% Chrysanthellum indicum, 1% Cynara scolymus, 1% Vaccinium myrtillus, 0.001% piperine) to form C9 also resulted in a strong reduction in glycemia to levels almost similar to the level of mice without diabetes mellitus.

Carbohydrate tolerance

Table 11. Effect of compositions on carbohydrate tolerance after eight weeks of administration Compositions AUC (week 8 - week 0, in mg×min/dl)
(mean ± SEM) control without restrictions 10,954 ± 2000 equal consumption control 20 355 ± 6554 C1 3367 ± 3039 ^* C2 1901 ± 3949 ^* C3 -2418±3260 ^** C4 -11 537 ± 4168 ^** C5 3610 ± 2086 ^* C6 13,068 ± 3064 C7 8639 ± 2844 C8 -8889 ± 5436 ^** C9 -24,568 ± 10,440 ^**

Mean values ± SEM. control without restrictions, n equals 22; control of equal consumption, n equals 10; C1, n is 9; C2, n is 8; C3, n equals 10; C4, n is 8; C5, n is 9; C6, n equals 14; C7, n is 12; C8, n equals 10; C9, n equals 9. AUC: “area under the curve.” Composition compared to control, Student's t test for unpaired data,^* p less than 0.05,^** p less than 0.01.

Compositions C1, C2, C3, C4, C5, C8 and C9 caused a decrease in food intake. As a result, a group was created that consumed the same amount of food. The results show that calorie restriction had no effect on carbohydrate tolerance (comparing unrestricted versus equal intake controls). Compositions C1, C2, C3, C4, C5, C8 and C9 caused a statistically significant decrease in AUC, reflecting an improvement in carbohydrate tolerance. The most pronounced effect was obtained with combinations of C4 and C9.

Body mass

Table 12. Effect of compositions on body weight after eight weeks of administration Compositions Body mass
(in g, mean ± SEM) control without restrictions 36.9 ± 1.2 equal consumption control 30.6 ± 1.2 C1 33.9 ± 2.4 C2 37.4 ± 2.2 ^* C3 37.0 ± 2.7 ^* C4 36.3 ± 1.7 ^* C5 32.7 ± 1.5 C6 29.3 ± 1.8 ^*** C7 34.5 ± 1.6 C8 26.9 ± 2.0 C9 25 0 ± 2.1 ^*

Mean values ± SEM. control without restrictions, n equals 34; control according to food intake, n equals 10; C1, n is 9; C2, n is 9; C3, n is 9; C4, n is 8; C5, n is 9; C6, n equals 14; C7, n is 12; C8, n equals 10; C9, n is 9. Composition versus control, Student's t test for unpaired data, ^* p less than 0.05, ^** p less than 0.01, ^*** p less than 0.0001.

Compositions C1, C2, C3, C4, C5, C8 and C9 caused a decrease in food intake. As a result, a group was created that consumed the same amount of food. Results show that calorie restriction caused a statistically significant reduction in body weight (unrestricted control versus equal intake control, p less than 0.05). Compositions C1, C2, C3, C4, C5, C8 and C9 also caused a statistically significant reduction in body weight. While the C6 combination had no effect on fasting blood glucose, it produced a strong and statistically significant reduction in body weight, giving it an advantage in the prevention and treatment of obesity. Combination C9 has the most pronounced effect.

Synergistic effect

Additionally, the synergistic effect was assessed in accordance with the Colby SR method, described in the article “Calculation of the synergistic and antagonistic responses of herbicide combinations” Weeds, 1967, 15:20-22. This method was, in particular, used in the patent EP03812880. For each combination, the synergy coefficient was calculated. A coefficient greater than 1 indicates the presence of a synergistic effect. A coefficient less than 1 indicates the presence of antagonism. The calculations were carried out as follows:

Expected degree of effectiveness = A + B - (A × B / 100)

Synergy Factor (SF) = (1 × Observed Effectiveness (%))/Expected Effectiveness (%)

Calculations apply to the following combinations:

Calculation of SF for the combination 1% Chrysanthellum indicum/1% Cynara scolymus/1% Vaccinium myrtillus/0.1% piperine (composition C4), in which A is 1% Chrysanthellum indicum/1% Cynara scolymus/1% Vaccinium myrtillus (C6 ) and B is piperine (C5);

Calculation of SF for the combination 1% Chrysanthellum indicum/1% Cynara scolymus/1% Vaccinium myrtillus/0.001% piperine/0.6% Olea europaea (composition C9), in which A is 1% Chrysanthellum indicum/1% Cynara scolymus/1 % Vaccinium myrtillus/0.001% piperine (composition C7) and B is 0.6% Olea europaea (composition C8).

Table 13 below shows the results for C6 + C5 = C4.

Table 13. Synergy coefficients for the combination C6 + C5 = C4 Observed degree of effectiveness (% of control group) Expected degree of effectiveness C4 Synergy coefficient C6 C5 C4 Fasting blood glucose after eight weeks of administration +2.6% -13.3% -35.3% -11.0% 3.20 Oral Carbohydrate Tolerance Test, AUC (Week 8 - Week 0) +9.5% -82.3% -156.7% -82.3% 1.94

The results presented in Table 13 are also illustrated in FIG. 2 and 3. The C4 composition of the invention demonstrated a strong synergistic effect on both fasting glycemia and carbohydrate tolerance (AUC).

Table 14 below shows the results for C7 + C8 = C9.

Table 14. Synergy coefficients for the combination C7 + C8 = C9 Observed degree of effectiveness (% of control group) Expected degree of effectiveness C9 Synergy coefficient C7 C8 C9 Fasting blood glucose after eight weeks of administration +5.7% -26.1% -59.4% -21.9% 2.71 Oral Carbohydrate Tolerance Test, AUC (Week 8 - Week 0) +4.6% -204.5% -388.7% -209.3% 1.86 Oral insulin sensitivity test, AUC (week 8 - week 0) -89.1% -53.2% -129.3% -94.9% 1.36

The results presented in Table 14 are also illustrated in FIG. 4, 5 and 6. The C9 composition of the invention demonstrated a strong synergistic effect on fasting glycemia, carbohydrate tolerance (AUC), and insulin sensitivity (AUC). This occurs even though C7 already has a strong and statistically significant effect on improving insulin sensitivity.

II.2 Tests B

Another composition, C10, was developed and tested in vivo: Chrysanthellum indicum (aerial part, dry extract) plus Cynara scolymus (leaf, dry extract) plus Vaccinium myrtillus (fruit, dry extract) plus Piper nigrum (fruit, dry extract) plus Olea europaea (leaves, dry extract).

More specifically, formulation C10 was tested in vivo in comparison with metformin, a standard therapeutic molecule for the treatment of type 2 diabetes, in the same model as for tests A (db/db mice). Using this predictive diabetic mouse model and when a C10 formulation was included directly in the rodent diet (1% Chrysanthellum indicum, 1% Cynara scolymus, 0.1% Vaccinium myrtillus, 0.001% Piper nigrum and 0.6% Olea europaea from the diet ) the "multi-purpose" effectiveness and wide-scale use of this composition can be guaranteed, since intravenous or intraperitoneal injections are limited to the small number of people carrying out this route of administration.

The duration of the experiment was seven weeks with a run-in period of one week, followed by six weeks of supplementation with composition C10. The male mice were six weeks old at the start of the experiment. Assessments were conducted immediately before supplementation (t = 0), once a week for some measures, and at the end of supplementation (t = 6 weeks).

Experimental assessments, in particular, related to:

Measuring food intake (weeks t1, t2, t3, t4 and t5);

Measuring body weight (weeks t0, t1, t2, t3, t4, t5, t6);

Measuring fat mass and lean tissue mass using MRI (magnetic resonance imaging; weeks t0 and t6);

Measuring fasting blood glucose (weeks t0, t1, t2, t3, t4, t5, t6);

Measuring the level of glycated hemoglobin (HbA1c; week t6);

Insulin sensitivity (test identical to that performed in Tests A; weeks t0 and t6);

Carbohydrate tolerance (test identical to that performed in Tests A; weeks t0 and t6);

Measuring triglyceride levels in the liver and serum (week t6);

Measurement of serum HDL cholesterol levels (week t6).

All of these estimates are generally familiar to those skilled in the art.

The results are presented below in Table 15: It is important to consider that the amount of metformin taken orally per day by the mice was approximately 26% greater than the total number of molecules present in the C10 formulation of the invention. In other words, the administered dose of metformin exceeded the dose of the total molecules present in the C10 composition.

Indicators Control Metformin C10 Food consumption (g/day) 9.6 ± 0.5 10.0 ± 0.6 10.7 ± 0.8 Body weight (g) 44.7 ± 2.0 40.8 ± 2.1 36.3 ± 1.7 ^a Adipose tissue mass (%) 26.1 ± 1.5 23.2 ± 1.7 19.1 ± 1.1 ^a Lean tissue mass (%) 17.0 ± 0.5 16.5 ± 0.5 16.5 ± 0.6 Fasting blood glucose (mg/dl) 437 ± 26 474 ± 24 160 ± 20 ^a,b HbA1c (%) 8.33 ± 0.53 7.59 ± 0.24 4.25 ± 0.20 ^a,b Area under the curve measured during an oral insulin sensitivity test (AUC, mg.min/dL) 50940 ± 2968 32778 ± 3166 14109 ± 1856 ^a,b Area under the curve measured during an oral carbohydrate tolerance test (AUC, mg.min/dL) 61050 ± 2757 65614 ± 1577 28823 ± 5333 ^a,b Serum triglycerides (mg/dL) 231.23 ± 18.29 256.23 ± 28.19 177.80 ± 19.60 ^b Triglycerides in the liver (μmol/mg tissue) 41.10 ± 1.80 38.39 ± 4.84 21.39 ± 2.21 ^a,b Cholesterol-HDL (mg/dl) 74.34 ± 4.10 99.21 ± 6.03 111.72 ± 8.39 ^a

Mean values ± SEM. Control, n equals 10; Metformin, n is 11; C10, n is 10. Student's t test for unpaired data. ^a : C10 compared to control, p less than 0.05. ^b : C10 versus metformin, p less than 0.05.

In FIG. 7 illustrates in more detail the effects of metformin and the C12 composition of the invention on insulin sensitivity.

The results illustrated in Table 15 demonstrate the extremely broad and unexpected effect of the C10 composition according to the invention on a group of metabolic syndrome risk factors - overweight, obesity, diabetes mellitus, non-alcoholic steatohepatitis (NASH) and cardiovascular diseases, namely:

Reducing body weight by reducing the mass of adipose tissue without affecting lean tissue;

Decrease in fasting glycemia and glycated hemoglobin levels;

Improving carbohydrate tolerance;

Improved insulin sensitivity;

Decrease in triglyceride levels in the liver and blood serum;

Increased levels of HDL cholesterol (“good cholesterol”).

The composition according to the invention not only leads to a reduction in the entire group of risk factors, but, moreover, to their correction to a level almost equivalent to that in mice without diabetes mellitus (healthy mice). In other words, the invention makes it possible to prevent the establishment and progression of type 2 diabetes mellitus.

In FIG. 8 and 9 illustrate the effects of the C10 composition of the invention on changes in body weight and fasting glycemia compared to a control group. The C10 composition leads to a sustainable limitation of body weight gain and prevents the development of glycemia.

In addition, its effects are far superior to those of metformin, which is the main antidiabetic drug (Ferrannini E et al. Eur Heart J 2015; Jun 10; example: GLUCOPHAGE®), and has been shown to have no side effects on circulating fats, unlike OCA (obeticholic acid , a farnesoid X receptor (FXR) agonist for the treatment of type 2 diabetes mellitus and NASH (non-alcoholic steatohepatitis) developed by INTERCEPT, which leads to a sharp decrease in circulating HDL cholesterol ("good cholesterol") (-0. 2 g/l, p less than 0.01). Quite the contrary, the composition according to the invention leads to an increase in HDL cholesterol levels by 50.3%.

In addition, the effect of the composition according to the invention exceeds the effect of the drug candidate developed by GENFIT, GFT 505 (Elafibranor), on the level of glycated hemoglobin (HbA1c), the main diagnostic indicator of type 2 diabetes mellitus and its pathological complications: composition C10, HbA1c is 4% compared to GFT 505, HbA1c is 6% for the highest dose (same db/db model and similar experimental design; Hanf R et al. Diab Vasc Dis Res 2014; 11:440-7). An additional 1% reduction in HbA1c reduces the risk of heart attack by 14%, cardiovascular disease by 37%, and amputation by 43%.

In addition, the composition according to the invention leads to a delay or even cessation of degeneration of pancreatic beta cells, which are the cells responsible for the secretion of insulin in response to increasing glycemia. In particular, type 2 diabetes mellitus causes gradual degradation of pancreatic beta cells, due in part to chronic hyperglycemia. This is reflected by an increase in insulin secretion in the early stages of the disease to reduce glycemia (increased insulinemia) followed by a gradual decrease in insulin secretion (decreased circulating insulinemia) due to the gradual destruction of beta cells (Leahy JL et al. J Clin Invest 1985;77:908-915) . The results obtained reliably confirm the effect of the C10 composition on pancreatic beta cells. In particular, after six weeks of supplementation, insulinemia in the group that was supplemented with the C10 composition according to the invention was higher compared to the control (27.40 ± 4.11 versus 9.64 ± 2.53 ng/ml, p less 0.01, respectively), reflecting a significantly different stage of diabetes progression: advanced diabetes mellitus with beta cell destruction in the control group (low insulinemia, high fasting glycemia, high HbA1c) and only slightly progressed diabetes mellitus or even no diabetes mellitus in the functional beta cell group, which was given the C10 composition according to the invention (high insulinemia, low glycemia, low HbA1c).

Thus, the invention fully satisfies the need for new preventive measures and therapeutic solutions, taking into account the multifactorial nature of cardio-metabolic disorders. This invention represents enormous progress in significantly eliminating the development of metabolic diseases and their pathological nature.

Molecular biological measurements of liver parameters were also performed. These assessments were performed after six weeks of administration and after the animals were sacrificed. In particular, the amounts of hepatic AMP-activated protein kinase (AMPK) were determined using Western blotting. AMPK is considered a metabolic sensor. AMPK, a versatile enzyme, is involved in the coordinated regulation of energy metabolism, food intake, and tissue sensitivity to various metabolic and hormonal signals. These properties thus ensure its role as a leading pharmacological target in the field of application for the treatment of metabolic diseases (diabetes mellitus, insulin resistance, obesity) and in the field of cardiology (coronary heart disease, complications due to diabetes mellitus) (Coughlan KA et al. Diabetes Metab Syndr Obes 2014;7:241-53). Increasing amounts of AMPK constitutes a strategy of primary interest in the identification and validation of therapies for type 2 diabetes mellitus and its complications (especially non-alcoholic steatohepatitis) and, more generally, pathologies related to metabolic disorders.

The results of liver studies show that the C10 composition according to the invention induces a significant increase in the amount of AMPK (control n is 9, 1.00 ± 0.22 compared to C10 n is 9, 1.96 ± 0.73, p less than 0.001, U test Mann-Whitney). These results are illustrated by Western blotting and shown in FIG. 10.

An experimental protocol, which can be performed by anyone skilled in the art, is given below.

AMFK

Protein isolation

50 mg of frozen tissue (liver) was placed in 20 volumes of NP-40 buffer (50 mM Tris HCl, pH: 7.4, 150 mM NaCl, 1 mM NaF, 1 mM Na ₃ VO ₄ , 1% Nonidet P-40, 0 .25% sodium deoxycholate) in the presence of 1 μl of protease inhibitor mixture (P8340, Sigma-Aldrich) and phosphatase inhibitor tablets (#88667 Thermo Fisher Scientific, USA). Tissues were homogenized on ice using a glass Potter homogenizer and then centrifuged at 14,000 g for 10 minutes at 4°C, after which the supernatant was separated. The protein content of the supernatant was quantified using a Bio-Rad DC kit (Bio-Rad, USA), and then all test samples were adjusted to the same standard concentration, then diluted a second time in 2X Laemmli buffer and heated at 90 °C for 3 minutes .

Blotting

For each test, the molecular weight scale and internal control were applied next to 15 μg of each test sample on a polyacrylamide gradient gel (4-15% Mini-PROTEAN® TGX Stain-Free™ gel, BioRad, USA). The gels were then exposed to a 300 V electric current for 18 to 20 minutes in electrophoresis buffer (25 mM Tris, 192 mM glycine, 0.1% sodium dodecyl sulfate (SDS)), and then the proteins were transferred to a polyvinylidene fluoride (PVDF) membrane. using a semi-fluidic transfer system (Transblot, Bio-Rad, USA) at a constant electrical current of 25 V and 2.5 A for 7 minutes. The membranes were then incubated in Tween and Tris buffered saline (TTBS: 50 mM Tris base pH: 7.5, 150 mM NaCl and 0.01% Tween 20) supplemented with 5% bovine serum albumin (BSA) for 1 hour at room temperature. The membranes were then washed with TTBS and then incubated overnight at 4°C with anti-AMPKα antibodies (D63G4, Cell Signaling, USA). After incubation, the membranes were washed again with TTBS and then exposed to horseradish peroxidase-conjugated anti-rabbit secondary antibody at a concentration of 1:3000 for one hour at room temperature. The membranes were then washed three times with TTBS and then exposed to a chemiluminescent solution (Clarity Western ECL; Bio-Rad, USA) for one minute. The membranes were then scanned on a Bio-Rad Chemidoc system and band intensities were measured using image analysis software (ImageLab V4.1, Bio-Rad, USA).

II.3 Test C

One of the major drawbacks of many therapeutic molecules currently commercially available is their acute hypoglycemic effect with a high risk of hypoglycemia. To test the acute hypoglycemic effect of the C10 composition according to the invention, the composition was administered by force feeding (400 mg/kg) to six healthy C57BL/6N mice. Glycemia was measured by tail biopsy before and after force feeding (minutes t0, t15, minute t30). Additionally, in a crossover protocol, the same mice were force-fed saline with glycemic measurements (minutes t0, t15, minute t30). The results presented in Fig. 11 demonstrate the absence of acute hypoglycemic effect of the C10 composition according to the invention.

II.4 Test D

Composition C11 of a single extract (hydroalcoholic dry extract) obtained from the powder of aerial parts of Chrysanthellum indicum, leaves of Cynara scolymus, fruits of Vaccinium myrtillus, fruits of Piper nigrum and leaves of Olea europaea (ratio 1/1/0.1/0.0001/0 ,6) were tested in the same diabetic mouse model used in tests A and B.

The duration of the experiment was 6 weeks with a run-in period of one week, followed by five weeks of supplementation with composition C11. The male mice were six weeks old at the start of the experiment. The assessment focused primarily on changes in body weight. Measurements were carried out immediately before the introduction of the additive (t equal to 0), once a week for some indicators and at the end of the introduction of the additive (t equal to 5 weeks). The composition was included in rodent food, and its proportion was 2.7% of the food (the same amount as for composition C10).

In FIG. 12 illustrates the effects of the C11 composition of the invention on body weight change compared to a control group. The C11 composition according to the invention results in a significant reduction in body weight gain.

I. Evaluation of the effectiveness of the composition in vitro

In vitro experiments were also performed to demonstrate the effects of compositions C10 and C11 according to the invention. A composition of the C12 combination of molecules according to the invention was also tested: dicaffeoylquinic acid plus apigenin-7-O-glucuronide plus monocaffeoylquinic acid plus oleuropein plus piperine (ratio 1/1/1/1/0.001).

Various enzymatic activity tests were performed to determine whether the compositions of the invention inhibit the strategic enzymes of carbohydrate and fat metabolism: alpha-glucosidase, dipeptidyl peptidase-IV (DPP-IV) and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-coenzyme A reductase). CoA). Targeting all of these strategic targets allows for global medical care for type 2 diabetes and its complications (diabetic foot, retinopathy, vision loss, nephropathy, cardiovascular episodes, non-alcoholic steatohepatitis, or NASH).

HMG-CoA reductase

To control circulating fats, it is necessary to reduce serum cholesterol concentrations by primarily targeting inhibition of cholesterol biosynthesis (inhibition of 3-hydroxy-3-methylglutaryl-coenzyme A reductase or HMG-CoA reductase). Pravastatin is one of the reference molecules for lowering LDL-cholesterol and total cholesterol levels. In particular, it is commercially available under the names ELISOR®, PRAVASTATIN MILAN, VASTEN®, PRAVASTATIN TEVA.

alpha-Glucosidase

Alpha-glucosidase is an enzyme that catalyzes the final step in the breakdown of carbohydrates. Alpha-glucosidase inhibitors (examples of drugs included in this category: GLUCOR®, DIASTABOL®) are used to reduce postprandial glycemia, which is considered a risk factor independent of macrovascular complications of diabetes mellitus (Kim JS et al. Biosci Biotechnol Biochem 2000; 64 :2458-61).

DPP-IV (dipeptidyl peptidase-IV)

DPP-IV inhibitors are considered one of the most promising treatment strategies for type 2 diabetes mellitus (von Geldern TW et al. Drug Development Research 2006; 67:627-42). DPP-IV rapidly inactivates the hormones GLP-1 (glucagon-like peptide-1) and GIP (glucose-dependent insulinotropic polypeptide), also known as incretins. Inhibition of DPP-IV allows, in particular, to enhance the action of GLP-1 and GIP, thus allowing the physiological stimulation of insulin secretion and inhibition of glucagon secretion by the pancreas, and therefore a decrease in glycemia. In addition, inhibition of DPP-IV promotes gastric emptying and causes a central anorexigenic effect. Thus, inhibition of DPP-IV provides an opportunity for better regulation of carbohydrate levels in individuals suffering from diabetes mellitus. Clinical studies have also shown that DPP-IV inhibitors are highly effective and well tolerated in the treatment of hyperglycemia in individuals with type 2 diabetes without causing weight gain or hypoglycemia (Green BD et al Diabetes Vasc Dis Res 2006;3:159-65). One of the standard inhibitors is diprotin A.

To evaluate the inhibitory potential of compositions C10, C11 and C12, the following experimental protocol was used:

Preparation of extracts, mixtures of extracts and mixtures of pure synthetic molecules

For HMG-CoA reductase and glucosidase inhibition tests, plant extracts were diluted in ultrapure water/ethanol mixtures, the proportions of which varied depending on the type of extract so as to obtain homogeneous solutions.

After homogenization, the test samples were centrifuged at 4000 rpm for 10 minutes and the supernatants were separated (centrifuge 1-15 Fisher Bioblock Scientif, Sigma-Aldrich®). The molecules were diluted in either 90/10 (v/v) ultrapure water/DMSO or 50/50 (v/v) ultrapure water/ethanol.

Mixtures of several plant extracts were diluted in a 50/50 (v/v) water/ethanol mixture and then sonicated for 15 minutes to homogenize the medium (using a Hielscher® UP50H ultrasonic transducer at maximum power). Finally, they were centrifuged at 4000 rpm for 10 minutes and the supernatants were separated (Fisher Bioblock Scientif Centrifuge 1-15, Sigma-Aldrich®).

For DPP-IV testing, test samples were diluted in either Trisma-HCl buffer (pH 8.0; 100 mM) or 90/10 (v/v) ultrapure water/DMSO.

HMG-CoA reductase inhibition test

This test measures the effectiveness of enzyme activity inhibition based on the decrease in absorbance over time at λ of 340 nm. The reaction mechanism is as follows:

HMG-CoA + 2NADP-H + 2H ⁺ →mevalonate + 2NADP ⁺ + CoA-SH

In this way, the disappearance of the substrate NADP-H is monitored (absorbance at 340 nm). Enzyme inhibition is indicated by a decrease in the change in absorbance at 340 nm over time relative to the control reaction without inhibitor (100% activity).

At the first stage, you need to prepare solutions from the Sigma® kit:

The 5x buffer supplied in the kit must be diluted to a concentration of 1x in ultrapure water;

NADP-H is dissolved by adding 1x buffer and then distributed into 100 µl tubes;

The initial enzyme solution is distributed in 25 μl aliquots.

Buffer and NADP-H were stored at −20°C, and enzyme aliquots were stored at −80°C until use.

The steps were carried out directly in 96-well microplates (NUNC96 or SARSTEDT96). Buffers, substrates, inhibitors and enzyme were added in a specific order (see below). For each microplate, it was necessary to prepare a blank, a control, and various test samples. Each element was prepared in duplicate (or even three):

blank sample preparation: 24 µl sample solvent, 12 µl HMG-CoA, 158 µl 1x buffer, 4 µl NADP-H; incubation at 37 °C, 3 to 5 min;

preparation of negative control: 24 μl of sample solvent, 12 μl of HMG-CoA, 158 μl of 1x buffer, 4 μl of NADP-H; incubation at 37 °C, 3 to 5 min; adding 2 μl of enzyme;

preparation of the test sample: 24 μl of a potential inhibitor sample, 12 μl of HMG-CoA, 158 μl of 1x buffer, 4 μl of NADP-H; incubation at 37 °C, 3 to 5 min; adding 2 µl of enzyme.

After plate preparation, reaction kinetics were monitored in a microplate reader (BMG Labtech) for 16 minutes, 25 seconds, with values read every 25 seconds at λ of 340 nm, for a total of 40 cycles. From these values the percentage of inhibition can be calculated.

α-Glucosidase Inhibition Test

α-Glucosidase inhibition tests were performed by monitoring spectrophotometry at λ of 405 nm for the formation of the yellow product para-nitrophenol (PNP) from the synthetic substrate: para-nitrophenyl-α-D-glucopyranoside (PNPg) according to the following hydrolysis reaction catalyzed α-glucosidases:

PNPg+ H ₂ O→PNP + glucose

Enzyme inhibition is indicated by a decrease in the change in absorbance at 405 nm over time relative to the control reaction without inhibitor (100% activity).

Recombinant human enzyme produced in Saccharomyces cerevisiae (maltase) (Sigma-Aldrich ®, G0660) was diluted in phosphate buffer (0.1 M, pH 6.8) to transfer from the original concentrated solution, theoretically containing 120 U/ml, to a solution with an activity of 1.6 U/ml used for tests.

During α-glucosidase inhibition tests, actions were performed directly in 96-well microplates (NUNC96 or SARSTEDT96). Buffers, substrates, inhibitors and enzyme were added in a specific order (see below). For each prepared microplate, a blank sample, a control, and various test samples are prepared. Each element is prepared in two repetitions (or even three):

Blank preparation: 100 µl phosphate buffer (0.1 M, pH 6.8), 20 µl sample solvent, incubation at 37 °C for 10 to 15 min followed by addition of 20 µl PNPg at a concentration of 2.5 mM (dissolved in phosphate buffer);

preparation of negative control: 100 µl phosphate buffer (0.1 M, pH 6.8), 20 µl sample solvent, 20 µl enzyme at a concentration of 1.6 units/ml, incubation at 37 °C for 10 to 15 min followed by the addition of 20 μl of PNPg at a concentration of 2.5 mM (dissolved in phosphate buffer);

preparation of the test sample: 100 μl of phosphate buffer (0.1 M, pH 6.8), 20 μl of a potential inhibitor sample, 20 μl of enzyme at a concentration of 1.6 units/ml, incubation at 37 °C for 10 to 15 min, followed by by adding 20 μl of PNPg at a concentration of 2.5 mM (dissolved in phosphate buffer).

After plate preparation, reaction kinetics were monitored in a microplate reader (BMG Labtech) for 30 minutes, with values read every 2 seconds at λ of 405 nm, for a total of 16 cycles. From these values the percentage of inhibition can be calculated.

DPP-IV inhibition test

DPP-IV inhibition tests are performed by monitoring spectrophotometry at 385 nm for the formation of the para-nitroaniline product from the synthetic substrate: Gly-L-Pro-p-nitroanilide according to the following DPP-IV catalyzed hydrolysis reaction:

Gly-L-Pro-para-nitroanilide + H ₂ O → para-nitroaniline + Gly-L-pro

Enzyme inhibition is indicated by a decrease in the change in absorbance at 385 nm over time relative to the control reaction without inhibitor (100% activity).

The concentration of the enzyme used was 0.045 U/mL (where U is μmol/min) and therefore needs to be diluted. Because the dilution was large, it was necessary to carry out two successive dilutions.

First dilution (obtaining a solution containing 5×10 ^-4 units/ml): (Vfinal: 450 µl; V(initial concentrated enzyme solution) = 5 µl)

Second dilution (obtaining a solution containing 5×10 ^-6 units/ml) (Vfinal: 1000 µl; V(D1)=10 µl); the working solution was solution D2.

The steps were carried out directly in 96-well microplates (NUNC96 or SARSTEDT96). Buffers, substrates, inhibitors and enzyme were added in a specific order (see below). For each prepared microplate, a blank, a control, and various test samples were prepared. Each element was prepared in duplicate (or even three):

preparation of a blank sample: 25 μl of sample, 25 μl of substrate (Gly-L-pro-p-nitroanilide dissolved in Tris-HCl buffer (100 mM, pH 8.0) at a concentration of 1.6 mM);

preparation of negative control: 25 μl of sample diluent, 25 μl of substrate, incubation at 37 °C, 10 min, 50 μl of enzyme at a concentration of 5 × 10 ^-6 units/ml (prepared in 100 mM Tris HCl buffer, pH 8, 0);

preparation of the test sample: 25 μl of a sample of a potential inhibitor, 25 μl of a substrate, incubation at 37 °C, 10 min, 50 μl of enzyme at a concentration of 5 × 10 ^-6 units/ml.

After plate preparation, reaction kinetics were monitored in a microplate reader (BMG Labtech) for 30 min, with values read every minute at λ of 385 nm, for a total of 31 cycles. From these values the percentage of inhibition can be calculated.

Calculation of percent inhibition

Typically, regardless of the test, for a given concentration of inhibitor, the equation curve was plotted: absorbance = f (time) (in minutes). In a similar manner, a curve was systematically plotted for controls (no inhibitor) and blank samples (no enzymes).

In the various curves obtained, only the linear sections at the beginning of the kinetics were retained, since they corresponded to the conditions for determining the initial velocities. Thus, the slope of uptake/minute was proportional to the rate of product appearance (glucuronidase and DPP-IV) or substrate disappearance (HMG-CoA reductase, negative slope).

Depending on the types of tests, the percentage of inhibition of various enzymes was calculated in different ways:

for tests for inhibition of HMG-CoA reductase and α-glucosidases, the calculation formula is identical: .

For the DPP4 inhibition test, the formula is slightly different since blank samples are taken into account:

.

Finally, the mean percent inhibition value was calculated for each inhibitor concentration, as well as the standard deviation of each value. If standard deviation values were greater than 10%, new tests were performed in duplicate until values less than 10% were obtained.

Summarized results obtained on the three enzymes for compositions C10 and C11 according to the invention are shown below in Table 16.

Product IC 50 ^a (µg/ml per well) % inhibition Alpha glucosidase
(human recombinant, Saccharomyces cerevisiae) DPP-IV ^b
(human recombinant DPP-IV expressed in SF9 cells) HMG-CoA reductase ^c (human enzyme catalytic domain - recombinant GST fusion protein expressed in E. coli) Acarbose 86.3 No inhibition No inhibition Diprotin A No inhibition 100 % No inhibition Pravastatin No inhibition No inhibition 100 % Composition C10 4.3 15.3% 48.5% Composition C11 4.9 29.3% 32.5%

^a IC 50: inhibition concentration for which the enzyme has only 50% of its activity. ^b Test sample concentration set to 1.25 mg/mL. ^c Test sample concentration set to 60 μg/mL.

Summarized results obtained on the three enzymes for the combination of molecules (C12 composition according to the invention) are shown below in Table 17.

Product IC 50 ^a (µg/ml per well) % inhibition Alpha glucosidase
(human recombinant, Saccharomyces cerevisiae) DPP-IV ^b
(human recombinant DPP-IV expressed in SF9 cells) HMG-CoA reductase ^c (human enzyme catalytic domain - recombinant GST fusion protein expressed in E. coli) Acarbose 86.3 No inhibition No inhibition Diprotin A No inhibition 89.3% No inhibition Pravastatin No inhibition No inhibition 100 % Composition C12 19.7 10.3% 30.1%

^a IC 50: inhibition concentration for which the enzyme has only 50% of its activity. ^b Test sample concentration set to 250 μg/mL. ^c Test sample concentration set to 60 μg/mL.

It was found that, in contrast to the standard inhibitory molecules for each enzyme [acarbose (drug), diprotin A (reference inhibitor), pravastatin (drug), respectively], only the C10, C11 and C12 compositions of the invention surprisingly inhibited three enzymes.

Due to the simultaneous effect on several regulatory processes, the compositions according to the invention represent advantageous prophylactic agents and therapeutic agents for the prevention and treatment of diabetes mellitus, dyslipidemias and their complications. Current therapeutic strategies consist of combining several drugs to reduce different risk factors individually. However, combining drugs can in some cases cause serious adverse reactions, for example, concomitant use of fibrates and statins increases the risk of myopathy (Denke MA J Manag Care Pharm 2003; 9:17-9). Thus, there is currently a real need for preventive measures and drugs whose “multi-target” mechanism of action has advantages in terms of suitability, tolerability and efficacy. Thus, the compositions according to the invention make it possible to reduce the risk of cardiovascular diseases, as well as to prevent and treat each dysfunction and/or its consequences separately.

Claims (59)

1. A composition for the prevention or treatment of carbohydrate and/or fat metabolism disorders in humans or animals, containing at least a mixture of molecules containing at least: - a single extract obtained from a mixture of at least two plants selected from Chrysanthellum indicum, Cynara scolymus, Vaccinium myrtillus, Olea europaea and Piper, and wherein the mixture of molecules further contains at least one of the following extracts: - one extract of Chrysanthellum indicum, if the mixture of plants of a single extract does not contain Chrysanthellum indicum, - one extract of Cynara scolymus, if the mixture of plants of a single extract does not contain Cynara scolymus, - one extract of Vaccinium myrtillus, if the mixture of plants of a single extract does not contain Vaccinium myrtillus, - piperine or Piper extract, if the plant mixture of a single extract does not contain Piper, and/or - one extract of Olea europaea, if the mixture of plants of a single extract does not contain Olea europaea, where the specified mixture of molecules contains: - at least one of apigenin-7-O-glucuronide, chrysantellin A, chrysantellin B, caffeic acid, luteolin, maritimethine, eriodictyol, isoocanin, apigenin, luteolin-7-O-glucoside, maritimein, marein, eriodictyol-7-O -glucoside, flavomarin, apigenin-8-C-α-L-arabinoside-6-C-β-D-glucoside (shaftoside), apigenin-6,8-C-di-β-D-glucopyranoside (vicenin-2) , And - at least one of dicaffeoylquinic acid, sulfomonocaffeoylquinic acid, luteolin, luteolin-7-O-glucoside, luteolin-7-O-glucuronide, apigenin-7-O-glucoside, cinaropicrin, and - at least one of monocaffeylquinic acid, delphinidin-3-galactoside, delphinidin-3-glucoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, cyanidin-3-glucoside, petunidin-3-galactoside, cyanidin-3-arabinoside , petunidin-3-glucoside, peonidin-3-galactoside, petunidin-3-arabinoside, peonidin-3-glucoside, malvidin-3-galactoside, malvidin-3-glucoside, malvidin-3-arabinoside, and - at least piperine, and - at least one of oleuropein, hydroxytyrosol; Where: - a single extract or extract of Chrysanthellum indicum is obtained using the whole plant and/or aerial parts of Chrysanthellum indicum; - a single extract or extract of Cynara scolymus is obtained using the whole plant and/or leaves of Cynara scolymus; - a single extract or extract of Vaccinium myrtillus is obtained using the whole plant and/or fruit of Vaccinium myrtillus; - a single extract or extract of Olea europaea is obtained using the fruits or leaves of Olea europaea; - a single extract or Piper extract is obtained using the fruits or leaves of Piper nigrum, Piper aduncum and/or Piper longum; wherein the extracts of Chrysanthellum indicum, Vaccinium myrtillus, Olea europaea and Piper are obtained using a water/ethanol mixture, the extract of Cynara scolymus is obtained using water; where the composition contains such an amount of the corresponding extract that allows the introduction of at least 0.00001 g of Chrysanthellum indicum extract, at least 0.00001 g of Cynara scolymus extract, at least 0.00001 g of Vaccinium myrtillus extract, at least 0.001 mg of piperine and at least 0.00001 g of Olea europaea extract per kg of body weight of the person to whom the composition is administered, per day. 2. The composition according to claim 1, characterized in that the mixture contains at least one dicaffeylquinic acid, apigenin-7-O-glucuronide, monocaffeylquinic acid, piperine and oleuropein. 3. The composition according to claim 1, characterized in that it also contains at least one additional element added additionally to the mixture of molecules, where said additional element is selected from: the following vitamins: B1, B2, B3, B5, B6, B8, B9, B12, C, A, D, E, K1 and K2; the following compounds: obeticholic acid, corosolic acid, polyunsaturated fatty acids of the omega-6 and/or omega-3 family, orotic acid, pangamic acid, para-aminobenzoic acid, amygdalin, beta-glucans, carnitine, dimethylglycine, imeglimine, isoflavones, L- arginine, oxytocin, pectin, pyridoxamine, resveratrol, viniferine, L-citrulline; the following trace elements and minerals: arsenic, boron, calcium, copper, iron, fluorine, iodine, lithium, manganese, magnesium, molybdenum, nickel, phosphorus, selenium, vanadium, zinc; the following microcomponents of non-essential nature: conjugated linolenic acid, lipoic acid, carotenoids, carnitine, choline, coenzyme Q10, phytosterols, polyphenols of the tannin and lignan family, taurine; fructooligosaccharides, galactooligosaccharides; bacteria that ferment lactic acid; yeast, such as red rice yeast (Monascus purpureus); mushrooms, such as grifola curly; products isolated from insects that are compatible with the food and pharmaceutical sectors; coating agents: for example hypromellose, microcrystalline cellulose, stearic acid, talc, sucrose, shellac, povidone, beeswax; flavoring agents: for example, natural blueberry flavoring or natural strawberry flavoring; acidulants such as malic acid; substances that prevent agglomeration: for example, silicon dioxide or magnesium stearate; thickeners such as xanthan gum, colloidal silicon dioxide, mono- and diglycerides of fatty acids; stabilizers such as calcium phosphate; emulsifiers such as soy lecithin; fillers such as corn starch; excipients, for example microcrystalline cellulose, magnesium stearate or dicalcium phosphate. 4. The composition according to claim 1, characterized in that it is in the form of a powder, gel, emulsion or liquid. 5. The composition according to claim 1, characterized in that it is made in the form of tablets, wafers, gelatin capsules, sticks, sachets, bottles, droppers or in injection form. 6. The composition according to claim 1, characterized in that the metabolic disorders of carbohydrates and/or fats are selected from diabetes mellitus type 1 and 2, non-alcoholic fatty liver disease, cardiovascular pathologies, pathologies associated with insulin resistance. 7. The composition according to claim 6, characterized in that non-alcoholic fatty liver disease is non-alcoholic steatohepatitis. 8. The composition according to claim 6, characterized in that cardiovascular pathologies are coronary cardiopathy, cerebrovascular diseases, peripheral arteriopathy, deep vein thrombosis. 9. The composition according to claim 6, characterized in that the pathology associated with insulin resistance is Alzheimer's disease. 10. Composition according to claim 1, characterized in that the disorder of carbohydrate and/or fat metabolism is dyslipidemia. 11. The composition according to claim 1, characterized in that the metabolic disorders of carbohydrates and/or fats are selected from obesity, overweight, metabolic syndrome and problems associated with blood pressure pathologies. 12. A method for the prevention or treatment of pathological disorders of the metabolism of carbohydrates and/or fats in humans or animals, including the administration of a composition according to any one of claims. 1-11 to humans or animals as food and/or medicine. 13. The method according to claim 12, characterized in that the disorder is diabetes mellitus type 1 and 2, and/or non-alcoholic fatty liver disease, and/or cardiovascular pathologies, and/or pathologies associated with insulin resistance . 14. The method according to claim 13, characterized in that non-alcoholic fatty liver disease is non-alcoholic steatohepatitis. 15. The method according to claim 13, characterized in that cardiovascular pathologies are coronary cardiopathy, cerebrovascular diseases, peripheral arteriopathy, deep vein thrombosis. 16. The method according to claim 13, characterized in that the pathology associated with insulin resistance is Alzheimer's disease. 17. Method according to any one of paragraphs. 12-16, characterized in that the composition is administered in combination with at least one antidiabetic therapeutic agent selected from biguanides, including metformin, dipeptidyl peptidase-IV (DPP-IV) inhibitors, glucagon-like peptide-1 (GLP-1) analogues, thiazolidinediones (TZD), sulfonylureas, short-acting and long-acting insulins, sodium-dependent glucose cotransporter-2 (SGLT2) inhibitors, glucosidase inhibitors (acarbose, miglitol, voglibose, peptides containing the alanine-proline or proline-alanine sequence), fibranor family molecules , such as elafibranor, or molecules targeting nuclear receptors, and in particular ROR receptors (α, β, γ) and Rev-Erb receptors (α, β). 18. The method according to claim 12, characterized in that the disorder is dyslipidemia. 19. The method according to claim 18, characterized in that the composition is administered in combination with a lipid-lowering therapeutic agent selected from: statins, fibrates, nicotinic acid, ion exchange resins, cholesterol absorption inhibitors, omega-3-polyunsaturated fatty acids, thiadenol and nuclear agonists farnesoid X receptor (FXR). 20. The method according to claim 12, characterized in that the disorder is selected from obesity, overweight, metabolic syndrome and problems associated with blood pressure pathologies.