EP4698200A1 - Pharmaceutical composition for the supportive treatment of diabetes mellitus - Google Patents

Abstract

The object of the invention is a pharmaceutical composition for the supportive treatment of diabetes and its use in the manufacturing dietary supplements for adjunctive therapy in the treatment of diabetes.

Description

Pharmaceutical composition for the supportive treatment of diabetes mellitus

The object of the invention is a pharmaceutical composition for the supportive treatment of diabetes and its use in the manufacturing dietary supplements for adjunctive therapy in the treatment of diabetes.

Deregulation of carbohydrate metabolism leading to hyperglycemia is the primary metabolic disorder in type 2 diabetes (T2D). T2D is often accompanied by complications such as cardiovascular disease, neuropathy, nephropathy, and diabetic retinopathy. Diabetes and its associated complications reduce patients' quality of life and generate a significant economic and social burden. Globally, the number of diabetes patients is estimated to increase to 592 million in 2035. In Europe, approximately 6% to 8% of the population suffers from diabetes, of which almost 90% suffer from T2D, making T2D the fastest-growing disease in Europe and the world. In T2D, there is excessive hepatic glucose production, insulin resistance, and insufficient insulin secretion by the pancreas. This contributes to an increased influx of fatty acids into the liver and muscle, causing the accumulation of lipid metabolites. Subsequently, this leads to dyslipidemia and nonalcoholic hepatic steatosis, ultimately result in increased cardiovascular risk. This process is accompanied by inflammation, which plays an important role in the pathogenesis of the atherosclerotic process in diabetes. Although oral antidiabetic drugs such as metformin are commonly used in the treatment of T2D, due to the side effects of these drugs, natural substances are being investigated to assist in the treatment of T2D, thereby reducing side effects. In this context, polyphenolic compounds have gained great popularity due to their widespread occurrence and antihyperglycemic effects, with minimal or no side effects. The mechanism of action of polyphenols in T2D is dependent on the type of compound and can differentially affect mechanisms (including insulin resistance, oxidative stress, inflammation, and dyslipidemia) involved in the development of T2D. For example, resveratrol, epigallocatechin gallate (EGCG) and quercetin have been shown to increase glucose uptake in muscle and adipocytes via GLUT4 translocation to the cell membrane, mainly through activation of the AMP-activated protein kinase (AMPK) pathway. It is well known that berries are rich in polyphenols and contain a wide range of phenolic molecules; hence, they are a good source of these compounds. Among others, clinical studies suggest positive effects of polyphenols from strawberries, cranberries, raspberries and aronia berries. The introduction of a 6-week dietary intervention in overweight and obese individuals, in the form of a daily intake of 333 mg of polyphenols from strawberries and cranberries, improved insulin sensitivity without affecting markers of inflammation and oxidative stress. In a study of pre-diabetic individuals, a single serving of 250 g of frozen raspberries with breakfast reduced maximum serum glucose and insulin levels and the area under the glucose and insulin curve in the post-meal response. However, no significant differences were found in inflammation and oxidative stress markers. A subsequent study showed that daily consumption of cranberry drinks for eight weeks did not improve insulin sensitivity but reduced isoprostane levels (an indicator of oxidative stress). Furthermore, it was found that in the group with elevated baseline C-reactive protein (CRP) levels, cranberry juice further reduced triglyceride levels. Also, daily consumption of 200 ml of aronia juice over three months effectively lowered fasting blood glucose levels in patients (6-17 years) with long-term T2D diabetes. Among other things, aronia had a beneficial effect on HbAlc, total cholesterol, and lipid levels.

Glucose and lipid metabolism are closely linked. Since metabolic diseases are often accompanied by disturbances in lipid and glucose metabolism, it seems reasonable to seek therapeutic strategies that target the underlying pathomechanisms of these metabolic pathways. Numerous pieces of research evidence suggest that the intake of polyphenols and their primary dietary sources may improve tissue insulin sensitivity and related diabetes risk factors such as hyperlipidemia, inflammation and oxidative stress. However, for people with T2D, it must also be taken into account that large amounts of fruit, due to their high sugar content, are not recommended in a diabetic diet. An additional problem is the varying composition of polyphenols in natural foods, depending on the source of origin and the weather conditions under which the plant grew. Therefore, it makes sense to develop a mixture of berry extracts standardized for their polyphenol content to maximize the positive effects of polyphenols on the mechanisms involved in T2D.

The object of the invention is a pharmaceutical composition for the supportive treatment of diabetes, which comprises:

- a mixture of elderberry, aronia, and blackcurrant extracts, - where a mixture of elderberry, aronia and blackcurrant fruit extract contains 10-40% anthocyanins and 20-80% polyphenols,

- the content of the extracts in the composition is 25-55% elderberry, 15-45% aronia and 10-35% blackcurrant.

The composition contains a pharmaceutical excipient or diluent, or carrier.

The composition is for oral administration, preferably as a tablet, capsule, or solution.

Composition, as defined above, is intended for adjunctive therapy with metformin for treating diabetes mellitus, preferably type II.

The composition has an anti-inflammatory application in a hyperglycemic state.

The composition has antioxidant applications in a hyperglycemic state.

The composition has a glucose-lowering application in a hyperglycemic state.

The composition has applications to improve lipid metabolism in a hyperglycemic state.

Composition as defined above for use in adjunctive therapy in pre-diabetic conditions.

Figure description:

Fig.l - shows the effect of metformin (1 mM), insulin (100 nM), composition (100 pg/mL) and composition (100 pg/mL) with metformin (1 mM) on the rate of glucose uptake by differentiated adipocytes of line 3T3-L1 using 2-NBDG. The graph shows the analysis results normalized in relevance to unstimulated cells.

Fig.2 - shows the effect of metformin (2 mM), composition (100 pg/mL) and composition (100 pg/mL) with metformin (2 mM) on the degree of phosphorylation of AMPK (A), ACC (B), HMG- CoA (C) proteins and SREBP1 protein levels (D) in HepG2 cells. The graphs show the results of densitometric analysis normalized in relevance -actin, presented as % of control.

Fig.3 - shows the effect of metformin (2 mM), composition (100 pg/mL) and composition (100 pg/mL) with metformin (2 mM) on cholesterol in HepG2 (A) and Thle-2 (B) and triglycerides in HepG2 (C) and Thle-2 (D). The graphs show the analysis results normalized in relevance to the 30 mM glucose control.

Fig-4 - shows the effect of composition on plasma glucose and insulin concentrations. The results of the analysis are presented as mean ± SD. The invention is illustrated by the following implementation examples, which do not constitute a limitation of the invention.

A COMPOSITION OF NATURAL PLANT EXTRACTS CONSISTING OF ELDERBERRY, BLACKCURRANT AND ARONIA EXTRACTS

The object of the invention is a composition comprising three natural extracts rich in polyphenolic compounds. The extracts were isolated from the fruits of the following plants: blackcurrant (Ribes nigrum L.), aronia (Aronia melanocarpd), and elderberry (Sambucus nigra L.). The raw materials used to produce the extracts come from Polish and European crops. The extracts used to manufacture the composition according to the invention are obtained by standard methods known in the state of the art.

The blackcurrant fruit extract used to implement the invention is characterized by a polyphenol content of 20% to 50% and an anthocyanin content of 10% to 35%. The aronia fruit extract used to implement the invention is characterized by a polyphenol content of 45% to 80%, anthocyanin content of 15 to 40%. The elderberry fruit extract used to implement the invention is characterized by a polyphenol content of 25% to 55% and an anthocyanin content of 15% to 40%. Advantageously, the extracts are in liquid, semi-solid or solid form.

According to the invention, the composition is a mixture of blackcurrant, aronia, and elderberry fruit extracts in the ratio of blackcurrant extract: aronia extract: elderberry extract of 10 to 35: 15 to 45: 25 to 55. The composition, a mixture of extracts, is standardised to contain anthocyanins from 10% to 40% and polyphenols from 20% to 80%.

The quantitative and qualitative lineup of the composition was selected based on research conducted on in vitro cellular models as part of the project entitled 'Development of diabetes treatment support products based on natural extracts rich in polyphenolic compounds'.

The combination of plant extracts in the proportions mentioned above allows for the preparation's base composition to be characterized by multidirectional effects, i.e., antiinflammatory, antioxidant, and glycemic -lowering effects in a hyperglycemic state. Example 1

A COMPOSITION OF NATURAL PLANT EXTRACTS CONSISTING OF ELDERBERRY, BLACKCURRANT AND ARONIA EXTRACTS

Evaluation of the effect of compositions on glucose uptake in an in vitro model of adipocytes differentiated from 3T3-L1 cells.

RESEARCH METHODOLOGY

Composition

A composition according to the invention containing a standardized extract of elderberry, blackcurrant and aronia. A stock solution of the composition at 50 mg/mL concentration was prepared in 40% DMSO and then brought to a 100 pg/mL concentration in the culture medium.

Cells used for experiments

Line 3T3-L1, differentiated from cells showing a fibroblast phenotype into morphologically and functionally mature adipocytes, was used as an in vitro model. The differentiated cells were cultured as a monolayer in DMEM medium supplemented with 10% FBS (fetal bovine serum), 100 U/ml penicillin, 100 pg/ml streptomycin and 0.25 pg/mL amphotericin B. Cells were used in the experiments after reaching 70% confluence.

Assessment of glucose uptake

This study aimed to measure glucose uptake by differentiated adipocytes of line 3T3-L1 using 2-NBDG (a fluorescent glucose analogue). 2-NBDG is a fluorescent glucose analogue used to monitor glucose uptake in living cells. Before the experiment, a 24-hour incubation in DMEM (high glucose) medium without FBS was used. After this time, the medium was changed to DMEM without added glucose and incubated for one hour. Cells were then stimulated with insulin (100 nM), metformin (1 mM) and a composition at 100 pg/mL (+/- 1 mM metformin), respectively. Subsequently, cells were washed and resuspended in PBS. Reading was performed at Ex/Em 485/535 nm. RESULTS

The composition at a 100 pg/mL concentration showed a synergistic effect with metformin, increasing glucose uptake by the cells (Fig. 1).

Example 2

A COMPOSITION OF NATURAL PLANT EXTRACTS CONSISTING OF ELDERBERRY, BLACKCURRANT AND ARONIA EXTRACTS

Evaluation of the effect of composition on fatty acid synthesis in an in vitro hepatocyte model of the HepG2 cell line.

RESEARCH METHODOLOGY

Composition

According to the invention, a composition contains a standardized extract of elderberry, aronia, and blackcurrant. A stock solution of the composition at 50 mg/mL concentration was prepared in 40% DMSO and then brought to a 100 pg/mL concentration in the culture medium.

Cells used for experiments

The HepG2 cell line was used as an in vitro hepatocyte model. Cells were cultured as a monolayer in MEM medium supplemented with 10% FBS (fetal bovine serum), 100 U/mL penicillin, 100 pg/mL streptomycin and 0.25 pg/mL amphotericin B. Cells were used in experiments after reaching 70% confluence.

Protein assessment

Cells of the HepG2 line were cultured in the presence of the composition at a concentration of 100 pg/mL (+/- 2 mM metformin, +/- 30 mM glucose, for 24h; +/- 100 nM insulin, for 10 min for selected conditions). Cell cultures were terminated after 24 h under the above conditions, followed by centrifugation and collection of the cell pellet. The Bradford method was measured in relevance to a BSA standard curve to assess the total protein concentration. Samples were brought to a concentration of 4 pg/mL, incubated with 1:1 Laemmli buffer (Biorad), and heated for 5 minutes at 95 °C. The samples and standard thus prepared were applied to a 10 per cent polyacrylamide gel (30 pg protein/path) and subjected to electrophoresis (120 V, two hours). After separation, electrotransfer was performed on PVDF membranes (70 min at 100 V) (Thermo Scientific). The membranes were then blocked in 5% BSA solution and incubated overnight, at four °C, with primary antibodies in TBST (Tab.l). The membranes were then washed 3x with TBST buffer and incubated with the secondary antibodies (Tab.l) in TBST, two hours, RT. After incubation, membranes were visualised using the Pierce™ ECL Western Blotting Substrate chemiluminescent kit (Thermo Scientific). The image obtained from the AZURE reader (Syngen Biotech) was subjected to densitometric analysis using Image Studio™ Lite Quantification Software.

Table 1. Primary and secondary antibodies with dilutions.

RESULTS

The mechanism of action of metformin is based on modulation of the AMP-activated protein kinase (AMPK)-dependent pathway. By activating AMPK in hepatocytes, the activity of acetyl-CoA carboxylase (ACC), as well as 3-hydroxy-3-methylglutaryl- coenzyme A (HMG-CoA) reductase, decreases, thereby reducing the number of enzymes required for fatty acid and cholesterol synthesis, respectively. In addition, AMPK decreases the expression of the transcription factor SREBP-1 (induced by insulin), which plays an important role in fatty acid metabolism and adipogenesis.

The results show a 36.9 % [95 % CI -37.3 - 111.1] increase in AMPK phosphorylation in the culture treated with 2mM metformin and a 346.2 % [95 % CI 272.0 - 420.4] increase in the culture with 100 pg/mL composition and 2mM metformin compared to the control culture without metformin. In addition, there is a 66% [95% CI 37.3 -94.7] increase in ACC phosphorylation in the culture treated with 2mM metformin and by 138.6% [95% CI 109.9 - 167.4] in the culture with 100 pg/mL composition and 2mM metformin compared to the control. The increase in HMG-CoA phosphorylation limits the amount of enzyme required for cholesterol synthesis. For 2 mM metformin, an increase in pHMG- CoA levels (and thus a decrease in activity) of 65.0 % [95 % CI 1.0-131.1] is observed, whereas for 100 pg/mL composition and 2mM metformin, an increase of 261.7 % [95 % CI 195.7-327.8] is observed relative to control. SREBP-1 protein is a key transcription factor in regulating de novo lipid synthesis in the liver. SREBP-1 protein levels increase under high glucose (30 mM) and high insulin (100 nM) conditions and decrease after metformin (2 mM) treatment. In cells incubated with the composition, SREBP1 protein levels decrease slightly, while they normalize to negative control levels in cells incubated with the composition administered in combination with metformin. (Fig.2).

Example 3

A COMPOSITION OF NATURAL PLANT EXTRACTS CONSISTING OF ELDERBERRY, BLACKCURRANT AND ARONIA EXTRACTS

Evaluation of the effects of composition on cholesterol and triglyceride levels in an in vitro model of HepG2 and Thle-2 cell line hepatocytes.

RESEARCH METHODOLOGY

Composition

According to the invention, a composition contains a standardized extract of elderberry, aronia, and blackcurrant. A stock solution of the composition at 50 mg/mL concentration was prepared in 40% DMSO and then brought to a 100 pg/mL concentration in the culture medium. Cells used for experiments

The HepG2 and Thle-2 cell lines were used as an in vitro hepatocyte model. HepG2 cells were cultured as a monolayer in MEM medium supplemented with 10% FBS (fetal bovine serum), 100 U/ml penicillin, 100 pg/ml streptomycin and 0.25 pg/mL amphotericin B, while Thle2 -in BEBM supplemented with 0.4% BPE, 0.1% Hydrocortisone, Retinoic Acid, Transferrin, Triiodothyronine, hEGF and 100 U/ml penicillin, 100 pg/ml streptomycin and 0.25 pg/mL amphotericin B. Cells were used in the experiments after reaching 70% confluence.

Evaluation of cholesterol and triglyceride levels in hepatocyte cultures in vitro

Determination of total cholesterol and triglycerides was performed in a 2D in vitro insulin resistance model in HepG2 and Thle-2 cells using luminescence-based assays. The Cholesterol/Cholesterol Ester-Glo™ assay measures cholesterol via cholesterol dehydrogenase, which links the presence of cholesterol to the production of NADH and the activation of proluciferin, which produces luminescence with luciferase. The test uses cholesterol esterase to measure cholesterol esters. The enzyme removes fatty acid from cholesterol esters to produce one molecule of cholesterol per ester molecule. The amount of cholesterol esters is determined by the difference in cholesterol measured in the esterase's absence (free cholesterol) and presence (total cholesterol).

The Triglyceride- Gio™ test measures triglyceride levels by measuring the glycerol released in an enzymatic reaction with lipase: one mole of glycerol per mole of triglyceride. Glycerol is measured in a coupled reaction that combines the production of NADH with the activation of proluciferin, which produces light using luciferase. The triglyceride amount is determined by the difference in the amount of glycerol measured in the absence (free glycerol) and presence (total glycerol) of lipase. Eipase converts triglycerides (TAG) into glycerol. Glycerol kinase and glycerol-3-phosphate dehydrogenase are used to generate NADH. In the presence of NADH, the reductase enzymatically reduces the proluciferin reductase substrate to luciferin. Luciferin is detected in the luciferase reaction using Ultra-Gio™ Luciferase and ATP, and the amount of luciferin generated is proportional to the amount of glycerol in the sample.

RESULTS In both the HepG2 and Thle-2 lines, the effects of metformin, as well as the composition and the composition with metformin, are similar. In the HepG2 line, there is a decrease in cholesterol levels to 60.5 ± 1.7; 54.8 ± 2.6, and 58.5±0.4 pM (from a baseline of 70.8 ± 0.1 pM in control) in cultures run with metformin, the composition, and the composition with metformin, respectively (Fig. 3A). In contrast, the Thle-2 line has cholesterol levels of 20.4 ±0.1; 22.4 ± 0.7 and 21.6 ±0.1 pM (with a baseline of 30.0 ±0.3 pM in the control) in cultures run with metformin, the composition, and the metformin composition, respectively (Fig. 3B).

In cells of the HepG2 line, triglyceride levels fall to 29.7 ±6.5 pM after metformin treatment, while they drop to 16.0 ±5.6 pM after composition treatment (from a baseline of 35.5 ±6.7 pM in control). Simultaneous application of the composition and metformin reduces triglyceride levels to 19.5 ±10.2 pM (Fig.3C). For the Thle-2 line, a slight decrease in triglyceride levels was observed after the use of metformin, i.e. from 69.7 ±1.0 pM in control to 66.1 ±3.3 pM. A significantly better effect was shown by the composition, lowering triglyceride levels to 46.6 ±2.1 pM and 39.3 ±6.7 pM in combination with metformin (Fig. 3D).

Example 4

ASSESSING THE EFFECT OF COMPOSITION ON THE LEVELS OF SELECTED PARAMETERS RELATED TO THE PATHOMECHANISM OF TYPE II DIABETES IN A MOUSE MODEL.

RESEARCH METHODOLOGY

PREPARATIONS

Fresh solutions were made each day, and then preparations were administered to the animals. Immediately before administration of the test preparation, the tubes were warmed to room temperature (approximately 21°C), and solutions were then made by dissolving the appropriate weight of preparation in saline. Animals received 1 ml of the preparation per 100 grams of body weight.

COMPOSITION - A composition according to the invention comprising a standardised extract of elderberry, aronia and blackcurrant.

METFORMIN - the standard oral antidiabetic drug. EXPERIMENTAL MODEL

Male db/db mice (BKS(D)-Lepr'® /JOrlRj) were used in the experiment as a model organism for type 2 diabetes research. The experiment consisted of daily intragastric administration of 0.9% NaCl solution/metformin/composition+metformin for six weeks.

Experimental groups: o CONTROL - animals receiving intragastric 0.9% NaCl solution (lml/100 g body weight/day) o METFORMIN - animals receiving metformin intragastrically at a concentration of 10 mg/ml in 0.9% NaCl solution (lml/100 g body weight/day) o COMPOSITION - animals receiving the polyphenol composition intragastrically at a concentration of 5 mg/ml in a 0.9% NaCl solution (lml/100 g body weight/day) o COMPOSITION + METFORMIN - animals receiving intragastrically the composition at a concentration of 5 mg/ml together with metformin at a concentration of 10 mg/ml in 0.9% NaCl solution (lml/100 g body weight/day)

After six weeks of treatment, all animals were sampled:

1) Whole blood into tubes with an anticoagulant (K3EDTA - tripotassium edetate). To obtain plasma, the blood was centrifuged (15 minutes, 3000 rpm, at 22°C).

2) Livers

After collection, the resulting plasma and livers were stored in liquid nitrogen for further analysis at - 80°C.

ASSESSMENT OF PROTEIN LEVELS

Protein levels in livers isolated from mice after six weeks of the experiment were assessed by Western Blot. Protein was isolated from 50 mg of tissue, which was mechanically minced and then resuspended in 600 pl of RIP A lysis buffer. Samples were lysed for 2 hours on ice while mixing every 15 min. on vortex. After 2 h, samples were centrifuged for 20 min. at 4°C, 16 000 g. The supernatant was collected into new tubes, and the precipitate was discarded. The Bradford method was used to assess the total protein concentration in relevance to the BSA standard curve. Samples were brought to a concentration of 4 pg/mL, incubated with 1:1 Laemmli buffer (BioRad), and heated for 5 minutes at 95 °C. The samples and standard thus prepared were applied to a 10- or 12- percent polyacrylamide gel (30 pg protein/well) and subjected to electrophoresis (120 V, 2 h). After separation, electrophoresis was performed on PVDF membranes (70 min at 100 V) (Thermo Scientific). The membranes were then blocked in 5% BSA solution and incubated overnight at 4 °C with primary antibodies in TBST (Table 1). The membranes were then washed 3x with TBST buffer and incubated with the secondary antibodies (Table 1) in TBST for 2h at room temperature. After incubation, membranes were visualised using the Pierce™ ECL Western Blotting Substrate chemiluminescent kit (Thermo Scientific). The image obtained from the AZURE reader (Syngen Biotech) was subjected to densitometric analysis using Image Studio™ Lite Quantification Software.

Table. 1. Primary and secondary antibodies used and dilutions. RESULTS

Table. 2. Effect of test formulations on phosphorylation rate or protein levels in the liver. Results of densitometric analysis were normalized to P-actin, and are presented as % of control ± SD.

*Pvalue for comparison against control (100%); for p<0.05 statistically significant results.

The results show a reduction in fasting glucose and insulin levels in mice treated with the composition administered with metformin (Fig.4).

Analysis of selected protein levels in mouse livers showed a significant decrease in ACC levels in the livers of mice supplemented with the composition (58.99 ± 22.89) and the composition in combination with metformin (56.14 ± 12.41 %). ACC catalyzes the production of malonyl-CoA from acetyl-CoA and affects lipid storage by regulating both fatty acid synthesis and oxidation. ACC promotes de novo fatty acid synthesis in lipogenic tissues such as the liver and adipose tissue. Phosphorylation of ACC inhibits the activity of this enzyme. The composition in combination with metformin significantly increased the phosphorylation of this enzyme (258.6 ± 62.98 % increase relative to control) compared to the effect of metformin alone and the composition alone. In addition, the low levels of ACC in the livers of mice supplemented with the composition in combination with metformin correlated with the observed reduction in SREBP1 levels (85.87 ± 22.74 %). In response to elevated insulin levels, SREBP-1 binds to and activates the transcription of lipogenesis genes, including fatty acid synthase and ACC. The livers of mice supplemented with the composition in combination with metformin also showed a significant reduction (52.91± 14.28 % decrease relative to control) in the activity (phosphorylation) of the p65 subunit of nuclear transcription factor kappa B (NF-KB). Activation of NF-KB leads to the transcription of genes encoding proteins associated with inflammation. The livers of mice supplemented with the composition in combination with metformin also showed a significant increase in SIRT1 levels (159.9 ± 40.67). SIRT1, an NAD(+)-dependent deacetylase in pancreatic P-cells positively regulates insulin secretion, protects cells from oxidative stress and inflammation, and plays a positive role in the metabolic pathway by modulating insulin signalling. SIRT1 also regulates adiponectin secretion, inflammation, glucose production, oxidative stress and mitochondrial function. Studies indicate that SIRT1 interacts with p65 to inhibit its transcriptional activity through deacetylation. SIRT1 also increases the expression of manganese-dependent superoxide dismutase (SOD2), one of the key regulators protecting cells from oxidative stress. The analysis showed an increase in SOD2 protein levels in the mouse livers of all groups tested, with the highest levels observed in the livers of mice supplemented with the composition.

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Claims

Patent claims

1. A pharmaceutical composition for the supportive treatment of diabetes mellitus, characterized by containing:

- a mixture of elderberry, aronia, and blackcurrant extracts,

- where a mixture of elderberry, aronia, and blackcurrant fruit extract contains anthocyanins 10-40% and polyphenols 20-80%,

- the content of the extracts in the composition is 25-55% elderberry, 15-45% aronia and 10-35% blackcurrant.

2. The composition, according to claim 1, is characterized by comprising a pharmaceutical excipient, or diluent, or carrier.

3. The composition according to claim 1, is characterized by being intended for oral administration, preferably in the form of a tablet, capsule, or solution.

4. The composition defined above is for use in adjunctive therapy with metformin for treating diabetes mellitus, preferably type II diabetes.

5. A composition according to claim 4, is characterized by its anti-inflammatory use in a hyperglycemic state.

6. The composition according to claim 4, is characterized by its antioxidant application in a hyperglycemic state.

7. The composition, according to claim 4, is characterized by its glucose-lowering application in a hyperglycemic state.

8. The composition, according to claim 4, is characterized by its use for improving lipid metabolism in a hyperglycemic state.

9. The composition as defined in claim 1 for use in adjunctive therapy in a pre-diabetic state.