WO2012138213A1

WO2012138213A1 - Reduction of vascular dysfunction with specific saccharides

Info

Publication number: WO2012138213A1
Application number: PCT/NL2011/050235
Authority: WO
Inventors: Klaske Van Norren; Niki Andreou Georgiou; Francina Jeannette Dijk
Original assignee: N.V. Nutricia
Priority date: 2011-04-08
Filing date: 2011-04-08
Publication date: 2012-10-11

Abstract

The invention pertains to the use of a preparation in the manufacture of a composition for the treatment and/or prevention of cardiovascular diseases, endothelial dysfunction and/or vascular dysfunction in human patients, said preparation comprising at least one saccharide selected from the group consisting of polyuronic acid having an average degree of polymerization [DP] between 2 and 250 saccharide units, arabinoxylan obtainable from rice bran, β-glucan obtainable from bakers yeast, and D-Ribose. The human patient is preferably suffering from HIV-infection or diabetes.

Description

Reduction of vascular dysfunction with specific saccharides

The present invention is in the field of endothelial dysfunction, and disorders such as vascular dysfunction and cardiovascular disorders associated therewith, particularly in a patient suffering from HIV infection or a diabetes patient.

BACKGROUND OF THE INVENTION

Vascular dysfunction is important in the pathogenesis of different diseases and pathogenic conditions like cardiovascular- diseases and diabetes. The vascular system in the human body is well described in the art. An important part of the system is the blood vessels that generally are divided in arteries and veins, dependent whether they transport blood to or from the heart. They vary in size from large (e.g. the aorta) to very small (capillaries). From an anatomical point of view larger blood vessels in general comprise as observed from the lumen side:

1. the tunica intima, that consists of a smooth (mono)layer of endothelial cells and a sub-endothelial layer that consists of a loose layer of connective tissue,

2. the tunica media, which consists of a layer of (innervated) smooth muscle cells and elastic fibers, and

3. the tunica adventitia which is composed of loosely woven collagen fibers, which are infiltrated by tiny lymphatic and blood vessels.

The endothelial cells in the tunica intima are in direct contact with blood and have a barrier function for the underlying tissue. This barrier function includes selective transport of components from blood to the underlying tissue and vice versa, and protection of the underlying tissue. The layer of endothelial cells can get disturbed by activation. If endothelial cells are activated, different families of adhesion molecules are expressed that facilitate the binding of leucocytes to the endothelial layer (adhesion molecules of the leucocytes bind to adhesion molecules of the endothelium) and in this way facilitate a local inflammatory response. This can eventually lead to e.g. local inflammatory responses and atherosclerotic plaque formation. Increased expression of adhesion molecules by the endothelium and leucocytes can be measured in vivo as an increase in levels of soluble forms of adhesion molecules (e.g. soluble ICAM). Increased levels of soluble adhesion molecules, e.g. sICAM or sVCAM have been identified as risk factors for ischemic events.

Vascular dysfunction is related to endothelial dysfunction with an increased expression of adhesion molecules like e.g. ICAM1. In fact, a widely used biochemical marker for endothelial dysfunction (activation) is the expression of ICAM- 1 on the endothelium, or the levels of sICAM-1.

Recently, highly active antiretroviral therapy (HAART) has reduced the risk of early death from opportunistic infections and extended the lifespan of people infected with the human immunodeficiency virus (HIV). However, in this HIV population there is still a risk of early death, but in this case from cardiovascular disease. Endothelial dysfunction is the most plausible link between HIV infection and atherosclerosis and cardiovascular diseases seen in these patients. The cause for this endothelial dysfunction probably lies in the HIV infection itself, not in the HAART therapy as thought before, although, some therapies might increase the effect. A widely used biochemical marker for endothelial dysfunction (activation) is the expression of ICAM- 1 or the levels of sICAM-1. Indeed, during HIV-1 infection, with or without HAART, ICAM-1 expression has been reported to be elevated. It has been suggested that both hyperlipidemia. hypercholesteremia and hyperglycemia can activate VCAM-1 expression on the vessel wall even in the absence of inflammation.

There is no consensus in the art on the working mechanisms leading to increased expression of endothelial adhesion molecules.

SUMMARY OF THE INVENTION

The inventors found that specific saccharides selected from the group consisting of polyuronic acid having an average degree of polymerization [DP] between 2 and 250 saccharide units, arabinoxylan obtainable from rice bran, β-glucan obtainable from bakers yeast, and D-Ribose, are capable of inhibiting NF-κΒ activation. The experimental data enclosed show that using in vitro models the Toll-like receptor induced, and the TNF-a induced NF-κΒ stimulation can be inhibited. The inventors have now found that these saccharides could be used advantageously for vascular dysfunction/cardiovascular disorders, associated with the NF-kB mediated

inflammation of endothelial cells. Without wishing to be tied down to any theory, the inventors believe that the specific nutritional oligosaccharides provided to the intestine also have systemic vascular effects. This is explained in more detail further below.

Outside the field of vascular dysfunction, WO 2010/147472 discloses a composition comprising polyuronic acid, arabinoxylan from rice bran, β-glucan from bakers yeast, or D-Ribose, or mixtures thereof, that achieves an inhibiting effect of NF-κΒ mediated virus replication. However, a link with endothelial dysfunction, and vascular dysfunction/cardiovascular disorders has not been suggested there.

Increased levels of soluble adhesion molecules, e.g. sICAM or sVCAM have been identified as risk factors for endothelial dysfunction, cardiovascular dysfunction, vascular dysfunction in diabetes e.g. hypertension, retinopathy, vascular damage in the extremities (diabetic feet) and in nephropathy, and HIV infected individuals.

In an aspect of the invention, the composition comprises at least one member selected from the group consisting of polyuronic acid having an average degree of

polymerization [DP] between 2 and 250 saccharide units, arabinoxylan obtainable from rice bran, β-glucan obtainable from bakers yeast, and D-Ribose, for use in the treatment and/or prevention of cardiovascular dysfunction in HIV infected patients, or for use in the treatment and/ prevention of hypertension, retinopathy, diabetic feet or

nephropathy in diabetes patients. In another aspect, the invention relates to a method for treating and/or preventing cardiovascular dysfunction in HIV-infected patients, and/or for treating and/or preventing hypertension, retinopathy, diabetic feet, or nephropathy in diabetes patients, by administering said HIV-infected or diabetes patient the composition as characterized herein.

In one aspect, the invention relates to a method for treating and/or preventing endothelial dysfunction, vascular dysfunction and/or cardiovascular dysfunction in a human patient, by administering said patient the composition as characterized herein. The patient is preferably suffering from HIV-infecttion or diabetes.

LIST OF FIGURES

Figure 1 : The effect of chain length of galacturonic acid oligosaccharides on NFkappaB inhibition. For inhibition by a polygalacturonic acid oligosaccharide a mimimal chain length of 2 galacturonic acid moieties is needed. The best effect is obtained with oligosaccharides comprising at least 3 galacturonic acid moieties;

Figure 2: NFkappaB response in HEK293 cells after 22.5 hr. incubation with AOS and NAC (A) or L-cysteine (B) with TNF-a for 2.5 hr. Upon correction for the contribution of NAC or cysteine , the curves C and D are obtained, showing that the components are additive;

Figure 3: Glutathione concentration at different NAC and AOS concentrations. The effect of the combination is more than the effect of the separate components over the complete curve, and is therefore synergistic.

DETAILED DESCRIPTION OF THE INVENTION

Through the nature of some of the nutritional carbohydrates as mentioned in example 1 , it can be assumed that an important site of action of these oligosaccharides will be in the intestine. Without being bound by theory, the inventors believe these

oligosaccharides might work via three different working mechanisms:

1) leucocytes, travelling freely through the body, sample the gut lumen and in this way can be influenced by the mentioned oligosaccharides, even if these compounds would not be detectable in the blood stream and on their turn have a different effect on the vessel wall;

2) the permeability of the gut changes under the influence of the

oligosaccharides changing inflammatory responses on the systemic vessel walls, but also changing oxidation of lipids influencing the quality of the vessel;

3) direct effects of (small amounts) of oligosaccharides from the gut reaching the vessel wall. Vascular dysfunction

'Vascular dysfunction' is clear terminology in the art, and is related to endothelial activation with an increased expression of adhesion molecules like e.g. ICAMl . In fact, a widely used biochemical marker for endothelial activation is the expression of ICAM-1 on the endothelium, or the levels of si CAM- 1. This activation is induced by chronic inflammation reactions in the patient. The skilled person has no difficulties to diagnose vascular dysfunction, using parameters widely available. It may also be referred to as 'endothelial dysfunction', underlying the targeted vascular dysfunction. One parameter is the carotid intima-media thickness (clMT). clMT can be measured by ultrasound at the carotid bifurcation or the internal carotid. Normally the clMT is in the range 0.75 +/- 0.06 mm. Improvement of vascular function is defined as a decrease in clMT progression or a decrease in clMT of 0.05mm during a treatment period of at least 1 month. (Hsue P, et all. Rapid progression of atherosclerosis at the carotid bifurcation is linked to inflammation in HIV-infected patients. CROI San Francisco, Feb 25-29, 2010, abstract 125).

Another parameter that could be used to measure improved vascular function is flow- mediated dilation (FMD), a functional assay for subclinical CVD risk. FMD measures the ability of brachial artery to dilate in response to endogenous release of nitric oxide following ischemic stress (Hsue P, et al. Inflammation is associated with endothelial dysfunction among individuals with treated and suppressed HIV infection. CROI, San Francisco, Feb. 25-29, 2010, abstract 708). Normal FMD values are in the range 8.2 +/- 5.0.

In the context of the present invention, the term 'vascular dysfunction' is preferably characterized by clMT and/or FMD, and is preferably understood to be defined by those clinical situations in which clMT and/or FMD are outside the normal range, i.e. the range therapeutically accepted as corresponding to normal conditions. In a study using patients with a history of stroke, the clMT was significantly higher in patients than in the control group (0.84 +/- 0.08 vs. 0.75 +/- 0.06 mm, P < 0.01), whereas brachial artery FMD% was lower in patients than in the controls (8.2 +/- 5.0 vs. 11.7 +/- 5.5%, P < 0.01). It is not necessary that both clMT and FMD are outside the normally acceptable levels, since it has been established in the art there does not need to be a significant correlation between CIMT and FMD% (r = -0.004, P = 0.94).

Without being bound by theory the inventors believe that each parameter characterizes a different aspect and stage of vascular dysfunction, therein supported by Kocak H. et al. Int Urol Nephrol. 2009;41(2):409-16. Relationship between carotid artery intima- media thickness and brachial artery flow-mediated dilation in peritoneal dialysis patients.

Cardiovascular disease is typically measured focussing on factors influencing aortic stiffness. This can be done by measuring the carotid- femoral Pulse Wave Velocity (PWV). Aortic PWV determined from a single measurement is shown to be strongly associated with the presence and extent of atherosclerosis, and that this measurement is highly related to cardiovascular risk as assessed by the standard Framingham equations. PWV can be determined with the use of an automatic device: the Complior (Colson), which allows an online pulse wave recording and automatic calculation of PWV with 2 transducers, 1 positioned at the base of the neck for the common carotid artery and the other over the femoral artery, as know to the person skilled in the art. In the overall population, optimal cutoff value of PWV to detect the presence of arteriosclerosis was 13 m/s with the following performance: 62% sensitivity, 67% specificity, 39% positive predictive value, and 84% negative predictive value. In patients without arteriosclerosis with age range from 30 to 74 years, optimal cut-off value of PWV to detect patients with high 10-year cardiovascular mortality risk was 13 m/s.

Above, the terms 'arabinoxylan from rice bran' and 'β-glucan from bakers yeast' are understood as 'arabinoxylan obtainable from rice bran' and 'β-glucan obtainable from bakers yeast', respectively. It is found that the saccharides from these sources have an effect on NF-κΒ activation, whereas arabinoxylan and beta-glucan from other sources do not. This is demonstrated in example 1. In a preferred embodiment, the composition comprises polyuronic acids, preferably digalacturonic acid, trigalacturonic acid and/or polygalacturonic acid, and cysteine equivalents, preferably N-acetyl cysteine (NAC), preferably having a weight ratio of polyuronic acid and cysteine equivalents between 10:1 and 1 : 10, more preferably 5 : 1 - 1 :5. Alternatively or additionally, the composition comprises 100 mg - 10 g of polyuronic acids, preferably pectin AOS per daily dosage. The same weight amounts and ratios are preferred in case of other saccharides defined in the description.

All saccharides preferably have chain length having 2 - 250 units. These can be derived from natural sources, if necessary by acid or enzymatic hydrolysis as known in the art.

In the context of the invention, the term "polyuronic acid" is understood to have a DP in the range of 2 to 250, more preferably a DP of 2 to 100, even more preferably a DP of 2 to 50, most preferably a DP of 2 to 20, thus also covering oligouronic acids. The uronic acid oligo- and polysaccharides are preferably galacturonic acid oligo- and polysaccharides (including digalacturonic acid, trigalacturonic acid). Most preferably, the degree of polymerisation is an average DP. In one embodiment, the composition comprises polygalacturonic acid. Most preferably, the composition comprises pectin hydro lysates, in the art typically addressed as pectin acid oligosaccharides (pAOS). Most preferably pectin (hydrolysate) is present in the form of or comprising digalacturonic acid, trigalacturonic acid, polygalacturonic acid. The uronic acid oligosaccharides, preferably pectin hydrolysates, are preferably obtainable or obtained by enzymatic digestion of pectin with pectin lyases, pectate lyase, endopolygalacturonase and/or pectinase.

In a preferred embodiment, the composition comprises omega-3 polyunsaturated fatty acids, preferably at least EPA, either alone or in combination with one or more cysteine equivalent(s).

Cysteine or source of cysteine

The inventors found that N-acetyl cysteine (NAC) is capable of blocking NF-KB activation. The mechanism behind this suppressive capacity has not been cleared, but it is hypothesized that it involves NAC's ability to decline oxidative stress indirectly via GSH level recovery. Oxidative stress has shown to promote NF-κΒ activation, via a mechanism that is still debated and probably cell-type specific. The inventors surprisingly found that both NF-kappaB activation and stimulation of glutathione formation was significantly less when one of the described oligosaccharides or a cysteine source, like cysteine or NAC, was provided alone, compared to when these two were combined. Combination with omega-3 fatty acids further increased the effect (table 3). The combination of NAC and AOS enhanced GSH production up to 278% compared with control. Data sets were compared with their baseline controls (0 mg/ml AOS in combination with its specific concentration of NAC or L-cysteine) and presented in example 2. High concentrations of AOS and NAC result in a relative stronger increase in GSH levels. From this, it is concluded that AOS and NAC have synergistic effects on the production of GSH in HEK293 cells.

The compositions provided comprise in addition to one or more oligosaccharides as described above a suitable amount of cysteine and/or source of cysteine. The phrase "source of cysteine" refers herein to all compounds that contain a biologically available cysteine, in any form, and is calculated as the amount of cysteine amino acid that is present in a compound, or can be derived from a compound in the body after ingestion, on a molar basis.

Hereinbelow "cysteine equivalent" refers to an amount of cysteine as such or to an amount of cysteine that is present in a source of cysteine. For example 100 mg NAC (N-acetylcysteine; MW= 163.2) is equivalent to 74 mg cysteine (MW 121.15). Thus 100 mg NAC is 74 mg cysteine equivalent. Similarly this can be applied to proteins or peptides. When a peptide (MW = xDalton) contains 3 cysteine amino acids (3yDalton), than 100 mg of this peptide is equivalent to 100x3Y/X mg cysteine. Thus lOOmg of this peptide is 300y/x mg cysteine equivalent.

Suitable sources of cysteine according to the invention are, for example, proteins in denatured and/or undenatured form such as milk proteins e.g. whey or casein proteins. Egg proteins are rich in cysteine and are therefore also suitable. Plant proteins such as pea, potato, soy and rice can also be used to provide cysteine. Also hydrolysates of these protein sources can be used or fractions enriched for cysteine rich proteins or peptides (e.g. as described in EP1201137). Furthermore, synthetic cysteine equivalents, e.g. derivatives of cysteine, such as cysteine, cysteine salts, N-acetylcysteine and/or diacetylcysteine can be used.

In yet another embodiment the compositions according to the invention comprise one or more compounds that stimulate glutathione levels, e.g. lipoic acid, pyruvate, oxaloacetate, oxaloaspartate, are capable in stimulating glutathione levels. Such glutathione level stimulating compounds may be used in addition to cysteine but also instead of cysteine. co-3 polyunsaturated fatty acids

The inventors found that co-3 polyunsaturated fatty acids are also capable of blocking NF-KB activation. This is demonstrated in the accompanying examples, where an effect is shown only in case such polyunsaturated fatty acids are combined with AOS and cysteine.

The compositions provided comprise in addition co-3 polyunsaturated fatty acids. The unsaturated fatty acid for a treatment in accordance with the invention is preferably selected from the group of C18-C26 co-3 polyunsaturated fatty acids. The composition preferably comprises one or more co-3 polyunsaturated fatty acids selected from the group consisting of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), eicosatetraenoic acid (ETA) and docosapentaenoic acid (DP A), most preferably at least DHA and/or EPA. Particularly good results have been achieved with a mixture comprising EPA. The daily dosage of C20-C26 co-3 polyunsaturated fatty acids can be between 200 mg - 2000 mg, preferably 200 - 2000 mg EPA and/or DHA, preferably 200 - 2000 mg EPA.

Further examples of co-3 polyunsaturated fatty acid, for use in accordance with the invention, include alpha- lino lenic acid and stearidonic acid, co-3 polyunsaturated fatty acids have been reported to be able to reduce ICAM expression. Uses of the invention

Diabetes:

One major cause of diabetic complications lays in vascular dysfunction. Recently more evidence is occurring that in e.g. hypertension , retinopathy, vascular damage in the extremities (diabetic feet) and in nephropathy, endothelial dysfunction and increased expression of adhesion molecules like ICAM-1 play an important role. It has been reported that high levels of glucose in combination with an inflammatory response like an increased level of cytokines increase ICAM-1 expression. Therefore, in one aspect, the invention is directed at the treatment of patients suffering from diabetes.

HIV patients:

The importance of chronic immune activation and its relationship with CD4+ T-cell depletion in HIV-infected individuals is increasingly recognized. In addition to rapid loss of plasma CD4+ T-cells, HIV infection is associated with a rapid loss of gut- associated CD4+ T-cells, impaired function of epithelial cells and alterations in the composition of the gut microbiota, leading to a loss of gastro -intestinal homeostasis and barrier function. As a consequence, bacteria or bacterial products such as

lipopolysaccharides (LPS) and peptidoglycans can translocate and enter the circulation, leading to chronic inflammation and immune activation. In HIV-1 infected individuals, including long-term non-progressive 'elite controllers', increased markers of bacterial translocation have been found and were correlated with parameters of immune activation and disease progression. Even in patients on highly-active antiretroviral therapy (HAART), markers of bacterial translocation correlate with immune activation and lower levels of CD4+ T-cell restoration. These data support the hypothesis that the gastro -intestinal tract plays a major role in HIV-induced chronic immune activation.

Chronic immune activation in HIV patients is believed to lead to activation of the vascular endothelium through NFk-B mediated inflammatory processes. Therefore the reductions found in NFk-B activation in the in vitro models found when using dietary fibers implies that these fibers can be used in the treatment of vascular dysfunction in many diseases suffering from chronic inflammation, including HIV. EXAMPLES

Example 1. Screening of oligosaccharides NFK-B inhibiting activity

Raftilose P95 (short chain FOS) No

D-Ribose Yes

β-D-glucan from barley No

β-D-glucan from baker's yeast Yes

Stachyose hydrate No

Trigalacturonic acid Yes

Vivinal-GOS No

Λ-carrageenan (irish moss) No

Example 2. Effect of oligosaccharides in combination with a cysteine source on NF- B inhibition

Methods: HEK293 cells were stably transfected with a NF-κΒ luciferase reporter gene construct. TNF-a was added to induce NF-κΒ activation. Citrus pectin oligosaccharide (pAOS) was added 18h prior to TNF-a addition (t=18h). As a cystein source, NAC was added at the same time as TNF-a. At t=20.5 the production of luciferase was measured by chemiluminescence.

Results: Both pAOS and NAC inhibited TNF-a-mediated NF-κΒ activation dose dependently, although the timeframe of activity differed per component (figure 2A and 2B). AOS -mediated NF-κΒ inhibition was optimal after an overnight pre-incubation period, while NAC-mediated inhibition was most effective after 2.5 hours. Upon correction for the contribution of NAC, the complete pAOS signal remained (figures 2c and 2D). This finding suggests that the components do not interact with each other, but inhibit NF-kB activation in a different, additive way.

Conclusions: The addition of NAC to pAOS results in a strongly improved inhibition of NF-KB. The lack of competition between the two components, together with different incubation times required, suggest different working mechanisms for both components. Example 3. Effect of oligosaccharides in combination with L-cysteine and/or EPA on NF-KB inhibition

Methods: HEK293 cells were stably transfected with a NFkappaB luciferase reporter gene construct. One day after cell plating EPA (0; 25 μΜ) was added at t=0h. pAOS (0; 1; 2; and 3 g/L) were added at t=96h and L-cysteine (0; 1 mM) at t=120h. NFKB activation was induced by 2.5h incubation with TNF-a. Luciferase activity was subsequently assessed at t=122.5h. As a control for cell viability cellular ATP levels were determined.

Results: The results are shown in Table 2, all standardized to a control experiment: NFKB activation in HEK293-NFKB-LUC cells stimulated with TNFa was inhibited by pAOS. L-cysteine increased the inhibition of pAOS. EPA alone did not inhibit NFKB, but when combined with pAOS and L-cysteine it further enhanced the NFKB inhibition. When combined, EPA, L-cysteine and pAOS showed an additional inhibition of NFKB activation. There was no competition between the components added. Each component reduced NFKB activation in a dose dependent manner. Cellular ATP levels remained unchanged.

Conclusion: The combination of pAOS, L-cysteine and EPA showed an additive inhibition of NFkappaB activation in vitro. The lack of competition between the three components, together with different incubation times required, suggest different working mechanisms for all three components.

Table 2. Inhibition of NFKB activation. Standardized to control.

*: p<0.05 compared to control. Example 4. Effect of oligosaccharides in combination with a cysteine source on glutathione

Methods: HEK293 cells were incubated with citrus pectin oligosaccharide (pAOS) for 18 h, then as a cysteine source, NAC was added and incubated for 2.5 h. After the incubation period, the cells were centrifuged (2000 rpm for 5 min). Medium was discarded and taken up into ice-cold perchloric acid (0.4 M) and re-suspended. Samples were then centrifuged for 10 min. at 13.000 rpm. Supernatant was collected and used for glutathione measurements. Total glutathione measurements were performed in 10 or 20 times diluted samples.

Results: Both pAOS and NAC stimulated glutathione production dose dependently (Figure 3), although the timeframe of activity differed per component. AOS-mediated glutathione production was optimal after an overnight pre-incubation period, while NAC-mediated glutathione production was most effective after 2.5 hours. Upon correction for the contribution of NAC, the complete pAOS signal remained. This finding suggests that the components do not interact with each other, but stimulate glutathione production in a different, additive way.

Conclusions: The addition of NAC to pAOS resulted in a synergistic effect on glutathione. Example 5. Effect of oligosaccharides in combination with a cystein source (L- cysteine) and EPA on glutathione

Methods: HEK293 cells were exposed to 25 uM EPA (t=0), one day after plating. HEK 293 cells were pre-exposed to 25 μΜ EPA for 96 hours. Then AOS (0, 1, 2, 3mg/ml) was added and after 24 hours (t=120 hours), TNFa (50 ng/ml) and 1 mM L-cysteine was added. At t=122.5h cells were resuspended in the media and glutathione assay was conducted. Total glutathione measurements were performed in 10 times diluted samples and levels were corrected for cell count.

Results: Cellular glutathione levels in HEK293-NFkB-Luc cells were slightly elevated in presence of pAOS. EPA (25 μΜ) alone did not affect GSH levels. When combined with AOS and L-cysteine, EPA increased GSH levels synergistically. By combining all three compounds, the increase of GSH levels was larger than the sum of the effects by the compounds alone. Conclusions: The addition of EPA to L-cysteine and pAOS resulted in a synergistic effect on glutathione levels.

Table 3. Cellular glutathione levels. Standardized to control at 1 mg/ml AOS.

*: p<0.05 compared to control.

Claims

1. Use of a preparation in the manufacture of a composition for the treatment and/or prevention of cardiovascular diseases and/or vascular dysfunction in human patients, said preparation comprising at least one saccharide selected from the group consisting of polyuronic acid having an average degree of polymerization [DP] between 2 and 250 saccharide units, arabinoxylan obtainable from rice bran, β-glucan obtainable from bakers yeast, and D-Ribose.

2. Use according to claim 1 wherein said human patient is suffering from HIV- infection or diabetes.

3. Use of a preparation in the manufacture of a composition for the treatment of endothelial dysfunction in HIV-infected patients and diabetes patients, said preparation comprising at least one saccharide selected from the group consisting of polyuronic acid having an average degree of polymerization [DP] between 2 and 250 saccharide units, arabinoxylan obtainable from rice bran, β-glucan obtainable from bakers yeast, and D-Ribose.

4. Use according to any of claims 1 -3, said polyuronic acid is derived from pectin.

5. Use according to any of claims 1-4, said polyuronic acid being in the form of or comprising digalacturonic acid, trigalacturonic acid and/or polygalacturonic acid.

6. Use according to any of the preceding claims, further comprising a cystein source selected from the group consisting of cystine, cysteine, cysteine salts, N- acetylcysteine and diacetylcysteine.

7. Use according to any of the preceding claims further comprising co-3 polyunsaturated fatty acids.

8. Use according to claim 7, comprising at least eicosapentaenoic acid (EPA).

9. A composition comprising at least one saccharide selected from the group consisting of polyuronic acid having an average degree of polymerization [DP] between 2 and 250 saccharide units, arabinoxylan obtainable from rice bran, β-glucan obtainable from bakers yeast, and D-Ribose for use in the treatment and/or prevention of cardiovascular diseases in human patients.

10. The composition for use according to claim 8, wherein said human patient is suffering from HIV-infection or diabetes.

11. A composition comprising at least one saccharide selected from the group consisting of polyuronic acid having an average degree of polymerization [DP] between 2 and 250 saccharide units, arabinoxylan from rice bran, β-glucan from bakers yeast, and D-Ribose, for use in the treatment of endothelial dysfunction in HIV- infected patients and diabetes patients.

12. A method for treating and/or preventing cardiovascular diseases, vascular dysfunction and/or endothelial dysfunction in a human patient, by administering said human patient a preparation comprising at least one saccharide selected from the group consisting of polyuronic acid having an average degree of polymerization [DP] between 2 and 250 saccharide units, arabinoxylan obtainable from rice bran, β-glucan obtainable from bakers yeast, and D-Ribose.

13. The method according to claim 12, wherein said human patient is suffering from HIV-infection or diabetes.

14. Use of a composition for the manufacture of a medicament for the treatment of a patient suffering from:

a. vascular dysfunction;

b. endothelial dysfunction; and/or

c. cardio vascular disease,

wherein the composition comprises at least one saccharide selected from the group consisting of polyuronic acid having an average degree of polymerization [DP] between 2 and 250 saccharide units, arabinoxylan obtainable from rice bran, β-glucan obtainable from bakers yeast, and D-Ribose.