OLIGOSACCHARIDES COMPOSITION FOR PREVENTING OR REDUCING THE RISK OF METABOLIC SYNDROME

Abstract

The present invention relates to the use of a galactooligosaccharide composition for preventing or reducing the risk of developing metabolic syndrome.

Claims

1. A composition comprising a mixture of galactooligosaccharides which comprises disaccharides Gal (ßI-3)-Glc; Gal (ßI-3)-Gal; Gal (ßI-6)-Gal; Gal (αI-6)-Gal; trisaccharides Gal (ßI-6)-Gal (ßI-4)-Glc; Gal (ßI-3)-Gal (ßI-4)-Glc; tetrasaccharide Gal (ßI-6)-Gal (ßI-6)-Gal (ßI-4)-Glc and pentasaccharide Gal (ßI-6)-Gal (ßI-6)-Gal (ßI-6)-Gal (ßI-4)-Glc for use in a method for preventing or reducing the risk of developing metabolic syndrome.

2.-14. (canceled)

Description

EXAMPLE 1

[0018] Freeze-dried powdered composition packaged in a ‘stick-pack’ and containing per 5.5 g final product:—

TABLE-US-00001 Galactooligosaccharide (GOS) mixture 2.75 g Lactose 1.40 g Monosaccharides (glucose, galactose) 0.64 g Drying aid 0.24 g Ash 0.23 g Moisture 0.19 g Protein 0.05 g

EXAMPLE 2

[0019] Syrup composition per 7.25 g finished product:—

TABLE-US-00002 Galactooligosaccharide (GOS) mixture 2.75 g Lactose 0.58 g Monosaccharides (glucose, galactose) 1.69 g Ash 0.23 g Moisture 1.95 g Protein 0.05 g

EXAMPLE 3

[0020] Effectiveness of galactooligosaccharides on metabolic syndrome risk factors in overweight adults

Study

[0021] A total of 45-50 human subjects with three or more risk factors associated with metabolic syndrome and its increased risk of cardiovascular disease were recruited. Equal numbers of men and women were included in the study cohort.

[0022] Metabolic syndrome factors used to select subjects included: insulin resistance (measured as increased ratio of fasting glucose (6-7 mmol/1) and insulin), high blood pressure, dyslipidaemia [low high density lipoprotein (HDL) cholesterol (<1 mmol/1), high triglyceride (>1.3 mmol/1)], waist circumference (>40 in men; >35 in women).

[0023] Further inclusion criteria were:— [0024] 18-65 years [0025] BMI >25 kg/m.sup.2 [0026] not having suffered a myocardial infarction/stroke or cancer in the past 12 months [0027] not diabetic (diagnosed or fasting glucose >7 mmol/l) or suffering from other endocrine disorders [0028] not suffering from chronic coronary, renal, bowel disease/gastrointestinal disorder or having a history of choleostatic liver or pancreatitis [0029] not on drug treatment for hyperlipidaemia, hypertension, inflammation, hypercoagulation or using drugs that affect intestinal motility or absorption [0030] no history of alcohol/drug abuse [0031] not planning or on a weight reducing regime [0032] not taking any dietary antioxidant or other phytochemical, prebiotics or probiotic supplements [0033] not pregnant, lactating, planning a pregnancy in the next 6 month or of child bearing potential and not using effective contraceptive precautions [0034] not taking antibiotics for the previous 1 month [0035] not anaemic (haemoglobin men >14 g/dl; women >11.5 g/dl) [0036] non smokers

[0037] Volunteers who met the inclusion criteria were asked to attend a screening session during which a fasting (12 hours) blood sample was taken and their BMI, waist circumference and blood pressure measured. The screening blood sample (˜10 ml) was analysed at the Royal Berkshire Hospital for total cholesterol (TC), HDL cholesterol, triacylglycerol, glucose and insulin. Individuals who were anaemic (Hb<14 g/dl male, 11.5 g/dl female) or who had ‘abnormal’ blood biochemistry based on the above analysis, were automatically excluded. The measurements were used to identify individuals at higher metabolic risk, who were then invited to participate in the study.

[0038] Treatment A or B was randomly assigned to each participant using an allocation ratio of 1:1 for the 2 study groups (including stratification for gender). Volunteers were required to attend the University for a total of 6 visits. The study was a randomised, controlled, double-blind crossover trial with Maltodextrin as the placebo. Volunteers were instructed to ingest the test product (GOS) and/or placebo daily for 12 weeks, with a 6 week washout period between. Both GOS and placebo were supplied in powder sachets (5.5 g) and volunteers were instructed to either sprinkle these over a bowl of cereal or add them to any drink, and ingest them daily. Habitual diet was assessed by pre-validated 4-day food diaries (2 weekend and 2 week days). At 0, 6 and 12 wk of intervention, volunteers visited the nutrition unit and samples and measurements were taken.

[0039] On each visit a fasting blood sample (˜20 ml) as taken and this was used to analyse a number of risk markers (all using commercially available kits). The markers studied were:

[0040] Lipid profile (total, low density lipoprotein (LDL) and HDL cholesterol, triglycerides and non-esterified fatty acids)

[0041] Insulin resistance derived from fasted measures of glucose and insulin ratio

[0042] Inflammatory/thrombotic biomarkers (including C-reactive protein, and IL6)

[0043] At 0, 12, 18 and 30 weeks a series of anthroprometric measurements (including weight, blood pressure and waist circumference) were taken in order to determine any changes.

Results

Baseline Characteristics of Subjects

[0044] The demographic characteristics of the study population are presented in Table 1.

TABLE-US-00003 TABLE 1 Baseline characteristics of subjects participating in the study. Characteristics Male (n = 16) Female (n = 29) Age (yr)  42.8 ± 12.1 46.4 ± 11.8 BMI (kg/m.sup.2) 30.7 ± 5.3 32.1 ± 6.3  Waist circumference (cm) 103.7 ± 11.0 99.2 ± 14.5 Fasting insulin (pmol/L)  66.3 ± 28.3 70.3 ± 30.6 Fasting glucose (mmol/L)  5.5 ± 0.8 5.2 ± 0.6 Systolic Blood Pressure (mmHg) 127.9 ± 10.1 125.9 ± 15.8  Diastolic Blood Pressure (mmHg) 80.9 ± 8.6 80.5 ± 10.0 Total Cholesterol (mmol/L)  6.3 ± 1.5 6.2 ± 1.2 HDL cholesterol (mmol/L)  1.3 ± 0.4 1.4 ± 0.3 TAG (mmol/L)  1.9 ± 0.9 1.4 ± 0.5

Effect on Colonic Microbiota

[0045] Daily consumption of 5.5 g of Bimuno (2.75 g active GOS) showed after 6 wk to result in a significant increase in the bifidobacterial population compared to both the Placebo (1 wk) (p<0.05) and baseline (p<0.05) levels (Table 2). After 12 wk of consumption, Bimuno intake resulted in significant increase in the populations of Bifidobacterium and Lactobacillus spp compared to Placebo (12 wk) (p<0.0001) and baseline (p<0.05). At the same time the levels of species of the Clostridium hystoliticum group and Desulfovibrio spp were significantly reduced compared to Placebo (12 wk) (p<0.0001) and baseline (p<0.05) (Table 2).

[0046] In terms of changes in the populations of Atopobium spp, C. coccoides/E. rectale, E. cylindroides, E. hallii, Clostridium cluster IX, F. prausnitzii cluster, beta-Proteobacteria, Bacteroides spp no significant effect was recorded either after the intake of Bimuno or Placebo during the 12 wks treatment period.

Effect on Biomarkers of Inflammation

[0047] Results on the levels of faecal sIgA (secretory immunoglobulin A), faecal calprotectin and blood inflammatory biomarkers (IL-6, CRP) during the study periods (Placebo, Bimuno) are shown in Table 3.

[0048] Daily intake of Bimuno for 12 weeks resulted in a significant reduction of both secretory IgA (p<0.05 vs Placebo; p<0.01 vs baseline) and calprotectin (p<0.01 vs placebo; p<0.05 vs baseline) (Table 3). At the same time a significant reaction in the blood levels of the pro-inflammatory cytokine IL-6 (p<0.05 vs Placebo; p<0.05 vs baseline) and in the inflammatory biomarker C-reactive protein (p<0.05 vs Placebo; p<0.05 vs baseline) was determined after daily intake of Bimuno for 12 weeks.

Effect on Metabolic Syndrome Risk Factors

[0049] Daily intake of Bimuno for 12 weeks resulted in a significant reduction in blood insulin levels (p<0.05 vs Placebo; p<0.01 vs baseline), blood triglycerides (p<0.05 vs placebo; p<0.05 vs baseline), total cholesterol (p<0.05 vs Placebo; p<0.05 vs baseline) and the ratio of total cholesterol over HDL cholesterol (p<0.05 vs Placebo; p<0.05 vs baseline) indicating an overall reduction of the risk of Metabolic syndrome (Table 4).

CONCLUSION

[0050] Supplementation with 5.5 g Bimuno (delivering 2.75 g of active GOS) in a population at risk of developing metabolic syndrome resulted in a significant change in the composition of their colonic microbiota by increasing the resident beneficial bacteria of Bifidobacterium genus and Lactobacillus genus, while decreasing the levels of detrimental bacteria such as those belonging to the C. hystoliticum subgroup and the sulphate reducing bacteria. This microbiota change resulted in an increase in the colonisation resistance in the gut reducing the colonic inflammation as seen through the reduction of the calprotectin levels. At the same time increased levels of sIgA would suggest a better barrier function of the epithelium which in combination with the reduced colonic inflammation could result in the reduction of the levels of inflammatory biomarkers in the host (IL-6, CRP).

[0051] This overall protection through the improvements in the composition of the colonic microbiota and the intestinal barrier function appears to have a beneficial effect in the levels of insulin, cholesterol and triglycerides that are known risk factors of metabolic syndrome.

TABLE-US-00004 TABLE 2 Changes in the numbers (Log.sub.10) of the various bacterial groups monitored during the study periods (Placebo, Bimuno), as determined by fluorescent in situ hybridisation (FISH) Placebo 5.5 g Bimuno Wk6 Wk12 Wk6 Wk12 Bifidobacterium spp  0.17 ± 0.13  0.3 ± 0.19  0.78 ± 0.23*  1.13 ± 0.29* Lactobacillus- −0.04 ± 0.09 −0.12 ± 0.18  0.24 ± 0.15  0.43 ± 0.22* Enterococcus spp C. hystoliticum group  0.15 ± 0.11  0.23 ± 0.07  0.12 ± 0.21 −0.61 ± 0.24* Desulfovibrio spp  0.02 ± 0.09 −0.03 ± 0.11 −0.04 ± 0.07 −0.63 ± 0.17* *significantly different from Baseline (p < 0.05)

TABLE-US-00005 TABLE 3 Changes in the levels of the faecal and blood biomarkers of inflammation during the study periods (Placebo, Bimuno) Placebo 5.5 g Bimuno Wk6 Wk12 Wk6 Wk12 sIgA (ug/g faeces) NA −230 ± 243  NA  902 ± 214* Calprotectin (ug/g 2.57 ± 4.03 2.58 ± 3.28 −2.91 ± 3.97  −9.61 ± 3.27* faeces) IL-6 (ng/ml) NA  7.97 ± 13.01 NA −33.34 ± 12.9* C-Reactive protein 0.46 ± 0.42 0.75 ± 0.40  0.66 ± 0.39  −1.56 ± 0.41* (ng/ml) *significantly different from Baseline (p < 0.05)

TABLE-US-00006 TABLE 4 Changes in the levels of insulin, TAG and cholesterol after 12 wk supplementation of Placebo or Bimuno Placebo 5.5 g Bimuno Wk6 Wk12 Wk6 Wk12 Insulin (pmol/L) 5.15 ± 3.14 7.42 ± 2.89 −0.02 ± 0.07 −10.37 ± 3.04* Triglycerides 0.09 ± 0.08 −0.03 ± 0.09  −0.08 ± 0.09  −0.79 ± 0.11* (mmol/L) Total Cholesterol 0.14 ± 0.11 0.05 ± 0.06 −0.10 ± 0.08  −0.39 ± 0.12* (mmol/L) Total Cholesterol: 0.01 ± 0.04 −0.06 ± 0.09  −0.19 ± 0.08 −0.44 ± 0.1* HDL *significantly different from Baseline (p < 0.05)