IE20060710A1

IE20060710A1 - Use of polyanhydroglucronic acid

Info

Publication number: IE20060710A1
Application number: IE20060710A
Authority: IE
Inventors: Anthony Richardson; Keith Real; Jiri Briestensky; Ivan Santar; Vladimir Semecky; Petr Nachtigal
Original assignee: Alltracel Dev Services Ltd
Priority date: 2005-09-28
Filing date: 2006-09-28
Publication date: 2007-07-25
Also published as: WO2007036920A3; WO2007036920A2

Abstract

The present invention provides a composition comprising a first component selected from a biocompatible anionic polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof, and an intermolecular complex thereof, and a second component comprising an anti-lipemic agent. The present invention also provides for use of a biocompatible anionic polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof or an intermolecular complex thereof, in the preparation of a medicament for the treatment of inflammation and use of a biocompatible anionic polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof or an intermolecular complex thereof, in the preparation of a medicament for maintaining a patient's blood glucose concentration in a physiological range. <Figure 4>

Description

The invention relates to use of polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof or an intermolecular complex thereof and a composition containing 5 PAGA, a salt thereof, a copolymer thereof and an intermolecular complex thereof.

Introduction Cardiovascular disease Cardiovascular disease (CVD) still remains the leading cause >f death and disability in the western world. The National Cholesterol Education Program (NCEP) identified elevated LDL (low density lipoprotein) as a major cause of CVD (JAMA 2001; 285 (19), 2486). The World Health Organization (WHO) has identified high cholesterol as the leading risk factor for CVD in the developed world. Approaches to reducing the risk of CVD include a combined strategy of interventions directed at high risk subjects coupled with a general population wide approach aimed primarily at lowering plasma low density lipoprotein cholesterol (LDL-C) and raising high density lipoprotein cholesterol (HDL-C) levels.

Cholesterol Plasma cholesterol is derived from two sources, endogenous (hepatic and extra-hepatic synthesis of cholesterol) and exogenous (intestinal absorption of dietary and biliary cholesterol). In order to eliminate cholesterol from the body, regardless of its genesis, it has to be excreted directly into bile or converted into bile acids and subsequently prevented from being re-absorbed to allow for elimination via the stools. A decrease in LDL of 10% in men is associated with a decrease in the risk of cardiovascular disease by 50% at the age of 40, by 40% at 50, by 30% at 60, by 20% at 70 and over (BMJ 1994;308:367-372).

ALPEI2/C •2Cholesterol although needed in the body can injure blood vessels and cause heart attacks and stroke when present in excessive amounts. The body needs cholesterol for digesting dietary fats, making hormones, building cell walls, and other important processes.

Lipoproteins Lipoproteins are particles that carry lipids and proteins in the blood and gut. There are five classes of lipoproteins: 1. Chylomicrons, which are mostly triglycerides and are secreted from the gut. 2. Very low-density lipoprotein (VLDL), which are mostly triglyceride and are secreted from the liver and gut. 3. Intermediate-density lipoprotein (IDL), which are mostly triglyceride and cholesterol and are secreted from the liver. 4. Low-density lipoprotein (LDL), which are mostly cholesterol and are secreted from the liver.

. High-density lipoprotein (HDL), which are mostly protein and are secreted from the liver and gut.

Two types of lipoproteins and their quantity in the blood are main factors in heart disease risk: LDL is ‘bad’ cholesterol and is the main cause of harmful fatty buildup in arteries. The higher the LDL cholesterol concentration in the blood, the greater the heart disease risk. HDL is ‘good’ cholesterol and helps prevent a cholesterol buildup in blood vessels. Low HDL levels increase heart disease risk.

Too much of this circulating cholesterol can injure arteries which can lead to accumulation of cholesterol-laden plaque in vessel linings, a condition called atherosclerosis.

Atherosclerosis 3ALPEI2/C The development of atherosclerotic lesions requires a complex interplay between mononuclear cells, endothelium, vascular smooth muscle, growth factors, and cytokines (Ross, 1999). Endothelial dysfunction followed by monocyte rolling and adhesion to the vascular endothelial lining and subsequent diapedesis are not only the first steps, but also seem to be crucial events in the atherosclerotic process (Nakashima et al., 1994). Several studies have demonstrated localized expression of leukocyte adhesion molecules in atherosclerotic lesions and plaques. They appear to regulate different stages of leukocyte migration at inflammatory sites in a multi-step process (Springer, 1994). Members of the immunoglobulin superfamily of endothelial adhesion molecules, vascular cell adhesion molecule (VCAM-1) and intercellular cell adhesion molecule (ICAM-I), strongly participate in leukocyte adhesion to the endothelium. VCAM-1 is highly expressed on endothelia prone to develop atherosclerosis in such atherosclerotic models as apoE ‘ mice, LDL receptor- deficient mice (LDLR') mice, and rabbits fed with an atherogenic diet (liyama et al., 1999; Nakashima et al., 1998). ICAM-I is expressed strongly on the endothelium overlying atheromatous plaque in human coronary and carotid arteries (DeGraba, 1997), hypercholesterolaemic rabbits (liyama et al., 1999), and apoE' (Nakashima et al., 1998) and LDLR ' mice (liyama et ah, 1999), although it is expressed in virtually all endothelial cells. One of the most significant advances in drug therapy during the twentieth century was the development of the statin class of drugs. These agents inhibit the activity of 3-hydroxy-3-methyIglutaryl coenzyme A reductase (HMG-CoA) as well as induce up-regulation of LDL receptors on the cell surface (Vaughan et al., 1996). In addition, a growing body of evidence suggests that statins exert beneficial vascular effects that are independent of their cholesterol lowering potencies (Farmer, 2000; LaRosa, 2001).

Cholesterol lowering materials Cholesterol levels may be decreased by several materials. These include statins, sterol/stanol, bile acid sequestrants, CETP (cholesteryl ester transfer protein) inhibitors, fibrates, niacin, dietary fibre etc. (i) Statins ALPEI2/C Statins target hepatocytes and inhibit HMG-CoA reductase (3-hydroxy-3-methyl-glutaryl coenzyme A reductase, E.C.l .1.1.34), the enzyme that converts HMG-CoA into mevalonic acid, a cholesterol precursor. The change in conformation at the active site makes these drugs very effective and specific. Binding of statins to HMGCoA reductase is reversible, and their affinity for the enzyme is in the nanomolar range, as compared to the natural substrate, which has micromolar affinity. The reduction of cholesterol in hepatocytes leads to the increase of hepatic LDL receptors that determine the reduction of circulating LDL and of its precursors (intermediate density - IDL and very low densityVLDL lipoproteins). All statins reduce LDL cholesterol non-linearly, dose-dependent, and after administration of a single daily dose. Efficacy on triglyceride reduction parallels LDL cholesterol reduction. Statins also inhibit hepatic synthesis of apolipoprotein B- 100, determining a reduction of the synthesis and secretion of triglyceride rich lipoproteins and an increase of receptors production for apolipoproteins B/E. Statins have a modest effect on HDL increase, and no influence on lipoprotein(s) concentration. (ii) Sterols Plant sterols are poorly absorbed in the intestine (0.4-3.5%), while absorption of plant stanols (0.02-0.3%) is even lower (for comparison, cholesterol absorption ranges between 35 and 70%). Plant stanols may also lower plant sterol absorption and vice versa. A reason for the low absorption of plant sterols and stanols might be that plant sterols and stanols are poorly esterified, possibly due to the low affinity of ACAT for these components. As merely esterified sterols are incorporated into chylomicrons, absorption of the unesterified plant sterols and stanols is consequently low. Different mechanisms have been suggested to explain the cholesterol-lowering activity of plant sterols and stanols. Firstly, plant sterols or stanols may displace cholesterol from mixed micelles, because they are more hydrophobic than cholesterol. This replacement causes a reduction of micellar cholesterol concentrations and consequently lowers cholesterol absorption. Furthermore, plant sterols or stanols might reduce the esterification rate of cholesterol in the enterocyte and consequently the amount of cholesterol excreted via the AI.PEI2/C chylomicrons. The effects of plant stanols on cholesterol absorption continue for at least several hours after ingestion. In response to the decreased cholesterol absorption, cholesterol synthesis increases. Also, LDL receptor mRNA and protein expression increases. This will not only increase clearance of LDL, but also of IDL. The higher LDL receptor expression and the higher endogenous cholesterol synthesis together result in an average reduction of LDL cholesterol of up to 14%. The decrease of total serum cholesterol is completely accounted for by a reduction in LDL. Plant sterols and stands have no effect on triacylglycerol or HDL cholesterol levels. In summary, stanols and/or sterols result in an LDL reduction of between 9 to 14%. (iii) Bile acid sequestrants (BASs) Bile acid sequestrants (BASs) were the first class of antihyperlipidemic drugs developed to lower LDL cholesterol levels. Two BASs, cholestyramine resin and colestipol, have been available for several years and have proven effective and safe as nonsystemic approaches to cholesterol reduction. However, tolerability and compliance issues, including complex dosage schedules, unpalatable formulations, and at times intolerable adverse effects, have limited the use of these traditional BASs.

To fully understand the lipid-lowering effects of BASs, it is essential to consider their effects on three hepatic enzymes: cholesterol 7a-hydroxylase, HMG-CoA reductase, and phosphatidic acid phosphatase. Routinely, bile acids returning to the liver through the enterohepatic pathway inhibit the activity of 7a-hydroxylase, the rate-limiting enzyme in bile acid synthesis. Conversely, the disturbance of enterohepatic recirculation activates this enzyme, leading to amplified conversion of intracellular cholesterol to bile acids.

The direct effect is a decrease in intracellular cholesterol stores that subsequently leads to a lowering of LDL cholesterol. Decreased intracellular cholesterol inversely affects the number of LDL cholesterol receptors and enhances the clearance of LDL cholesterol from the circulation; also, it stimulates the hepatic production of cholesterol by increasing the activity of HMG-CoA reductase, the rate-limiting enzyme in cholesterol synthesis. Despite this increase in cholesterol synthesis, however, total cholesterol levels do not -6A1.PEI2/C λΠ\ο increase because of the rapid shunting of cholesterol into the bile acid synthesis pathway. The activation of HMGCoA reductase may explain the success of combination therapy with BASs and HMG-CoA reductase inhibitors for primary hypercholesterolemia.

In addition to decreasing total cholesterol and LDL cholesterol, BASs can cause an undesirable increase in serum triglycerides. The increase is triggered by activation of phosphatidic acid phosphatase, an enzyme responsible for the conversion of glycerol phosphate to either triglycerides or phospholipids. Phosphatidic acid phosphatase is suppressed under the normal conditions of a functional enterohepatic recirculation system. However, upon interruption of the system by BASs, the increased activity of this enzyme leads to triglyceride production and changes in the size and content of very-lowdensity (VLD) lipoproteins. Increases in plasma triglyceride levels of 10-14% have been observed in patients receiving BAS therapy, but the magnitude of the increase may depend on the patient’s genetic predisposition to hypertriglyceridemia. Because of their potential to elevate serum triglycerides, BASs should not be used as monotherapy for patients with increased triglyceride levels. The triglyceride-elevating effects of BASs may be countered by the addition of agents known to decrease triglyceride synthesis, such as fibric acid derivatives (i.e., fenofibrate and gemfibrozil) and nicotinic acid. There are many examples of BASs but the two main commercially available ones are Welchol and chitosan. These generally lead to an LDL reduction of about 10%. (iv) Dietary ftbre During the last couple of decades, much attention has been given to the role of dietary fibres in the control of lipid and lipoprotein metabolism. Dietary fibres include a variety of plant substances, mainly nonstarch polysaccharides and lignins, which are resistant to digestion by digestive enzymes. They can be classified into two groups based on water solubility. Soluble fibres may lower plasma total cholesterol by a specific effect on LDL cholesterol. HDL cholesterol or triacylglycerol concentrations are in general not affected. Several mechanisms of action for the hypocholesterolemic effect of soluble fibres have been suggested that may depend on the type of fibre. Soluble fibres may ALPEI2/C increase the binding of bile acids in the intestinal lumen, which leads to a decreased enterohepatic circulation of bile acids and a subsequent increase in the hepatic conversion of cholesterol to bile acids (see above BAS section). Another suggested mechanism is that the increased viscosity of the food mass in the small intestine because of soluble fibres leads to the formation of a thick unstirred water layer, adjacent to the mucosa. This layer may act as a physical barrier to reduce the absorption of nutrients and bile acids. Increased intestinal contents supernatant viscosity is highly correlated with reduced plasma and liver cholesterol and reductions in cholesterol absorption in hamsters. Furthermore, soluble fibres may reduce the rate of glucose absorption, leading to a lower glycemic response and lower insulin concentrations. This latter may result in a reduced hepatic cholesterol synthesis. There are many examples of high viscosity soluble fibres. The main commercially available soluble fibres are psyllium, glucomannan and β-glucan. These generally lead to an LDL reduction of about 6 to 7%. (v) Cholesterol ester transfer protein (CETP) inhibitors The marked increase in HDL associated with human deficiency of cholesteryl ester transfer protein (CETP) has suggested CETP inhibition as a means of elevating HDL. Expression of CETP in transgenic mice under different metabolic settings has produced mixed results regarding its atherogenicity, whereas inhibition of endogenous CETP in rabbits has more consistently been antiatherogenic. With regard to human CETP mutations and the associated reduction in CETP levels, recent analysis of prospective data from the Honolulu Heart Study is consistent with the results of a previous study of Japanese subjects in concluding that CETP deficiency is protective when associated with HDL-C levels _60mg/dL.

Recently, a new CETP inhibitor, Torcetrapib, was tested in two studies in human subjects. In the first, the effect of CETP inhibition on plasma HDL levels was studied in healthy subjects. With the highest dose of 120 mg twice daily, given for 14 days, CETP activity decreased by 80%, while the CETP mass increased, apparently because the mechanism of action of Torcetrapib represents the shift of free CETP to HDL-bound ALPEI2/C form. With the above treatment, plasma LDL decreased by 42%, HDL-C increased by 91 %, as did apo A, and apo E, by 27 and 66%, respectively. In the second study, Torcetrapib was given to 19 normal subjects with low plasma HDL-C levels (<40 mg/dL), 9 of whom received atorvastatin, 20 mg/day (Brousseau et al. 2004). In subjects who received only 120 mg/day Torcetrapib for 4 weeks, HDL was increased by 46%; in those treated also with atorvastatin, the increase was 61%. A much higher rise in HDLC, 106%, occurred when the drug was given twice daily, but no atorvastatin. In six subjects treated with the two drugs, there was also a 17% decrease in LDL-C. In addition, Torcetrapib decreased the levels of small dense LDL and increased the concentration of large HDL to values seen in subjects with normolipidemia. (vi) Targeting absorption of cholesterol Ezetimibe is a US Food and Drug Administration-approved drug that targets the absorption of cholesterol in the intestine. The identification of this drug has also led to the elucidation of the dietary cholesterol receptor. Ezetimibe is efficacious as a plasma cholesterol-lowering agent as monotherapy, but its greatest utility seems to be as a combination with a low-dose statin, where it results in cholesterol lowering that is equivalent to using maximum-dose statins. It has a very favourable side-effect profile, as well as a lack of drug-drug interactions. Ezetimibe is a cholesterol-lowering agent that inhibits the intestinal absorption of cholesterol, presumably by interacting with a transporter responsible for the passage of cholesterol across the intestinal wall (vii) Niacin Nicotinic acid (niacin) lowers total and LDL cholesterol and raises HDL cholesterol. It also can lower triglycerides. Because the dose needed for treatment is about 100 times more than the recommended daily allowance for niacin and thus can potentially be toxic, the drug must be taken under a doctor’s care. Fibric acid derivatives such as gemfibrozil and fenofibrate can also increase HDL levels, but are used mainly to lower triglycerides.

AI.PH2/C Diabetes Mellitus Diabetes mellitus is a disease characterized by persistent hyperglycemia (high blood sugar levels), resulting either from inadequate secretion of the hormone insulin, an inadequate response of target cells to insulin, or a combination of these factors. Type 2 diabetes mellitus is due to a combination of defective insulin secretion and defective responsiveness to insulin (often termed reduced insulin sensitivity). In early stages the predominant abnormality is reduced insulin sensitivity, characterized by elevated levels of insulin in the blood. The initial defect of insulin secretion is subtle and initially involves only the earliest phase of insulin secretion. In the early stages, hyperglycemia can be reversed by a variety of measures and medications that improve insulin sensitivity or reduce glucose production by the liver, but as the disease progresses the impairment of insulin secretion worsens, and therapeutic replacement of insulin often becomes necessary.

There are many risks to health and these include cardiovascular disease, more than 65% of people with diabetes die from heart disease or stroke (American Diabetes Association). The complications are less common and less severe in people who have well-controlled blood sugar levels.

It is clear that a composition for lowering cholesterol levels and use of a composition for serum glucose management and/or for use as an anti-inflammatory agent would be very beneficial both therapeutically and commercially.

Statements of Invention According to the invention there is provided a composition which comprises a first component selected from a biocompatible anionic polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof, and an intermolecular complex thereof, and a second component that comprises an anti-lipemic agent.

ALPE12/C The second component may be selected from a sterol, an esterified and/or hydrogenated sterol, a stanol and a statin. Preferably, the second component may comprise a sterol. Alternatively, the second component maybe a satin such as a 3-Hydroxy-3methylglutaryl CoA (HMG CoA) reductase inhibitor.

The second component may be selected from one or more of the group consisting of: a cholesteryl ester transfer protein (CETP) inhibitor; a enterocyte cholesterol transporter inhibitor; nicotinic acid; niacin; a peroxisome proliferator-activated activator (PPAR) agonist; fish oil and soluble fibre such as psyllium, glucomannam, HPMC cellulose or glucan.

The composition may be suitable for oral administration. Preferably, the composition may be in the form of a tablet, pellet, capsule, granule or microsphere. Alternatively, the composition may be in a form suitable for incorporation into foods, beverages, nutraceutical or pharmaceuticals.

The first component may be derived from a starch, cellulose or gum, or is of microbial origin.

Preferably, the first component may comprise a microdispersed cellulose or derivative thereof.

The first component may be prepared by partial or complete hydrolysis and neutralisation of a PAGA containing material.

The first component may interact with biomolecules in the fluid media of the gastrointestinal tract.

The composition may further comprise at least one biocompatible biologically active substance. ΛΙ.ΡΕ 12/C The composition may further comprise at least one biologically acceptable adjuvant such as a pharmaceutically active adjuvant. The adjuvant maybe an antilipemic agent such as a phospholipid.

The invention also provides use of a biocompatible anionic polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof or an intermolecular complex thereof, in the preparation of a medicament for the treatment of inflammation, such as inflammation associated with atherosclerosis.

The invention further provides use of a biocompatible anionic polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof or an intermolecular complex thereof, in the preparation of a medicament for maintaining a patient’s blood glucose concentration in a physiological range. For example in patients with Type 2 Diabetes Mellitus.

The medicament maybe for oral administration. For example, the medicament maybe in the form of a tablet, pellet, capsule, granule or microsphere. Alternatively, the medicament may be in a form suitable for incorporation into foods, beverages, nutraceutical and pharmaceuticals.

The polyanhydroglucuronic acid, a salt thereof, a copolymer thereof and an intermolecular complex thereof may be derived from a starch, cellulose or gum, or may be of microbial origin.

The PAGA may comprise a microdispersed cellulose or derivative thereof.

The PAGA may be prepared by partial or complete hydrolysis and neutralisation of a PAGA containing material.

The PAGA may interact with biomolecules in the fluid media of the gastrointestinal tract.

ALPEI2/C The medicament may further comprise an antilipemic agent. For example, the antilipemic agent may be a sterol, stanol, statin, esterified and/or hydrogenated sterol.

The statin may be a 3-Hydroxy-3-methylgIutaryl CoA (HMG CoA) reductase inhibitor. Alternatively, the antilipemic agent may be selected from one or more of the group consisting of: a cholesteryl ester transfer protein (CETP) inhibitor; a enterocyte cholesterol transporter inhibitor; nicotinic acid; niacin; a peroxisome proliferatoractivated activator (PPAR) agonist; fish oil and soluble fibre such as psyllium, glucomannam, HPMC cellulose or glucan.

The medicament may include at least one biocompatible biologically active substance.

The medicament may include at least one biologically acceptable adjuvant such as a pharmaceutically active adjuvant. The adjuvant may be an antilipemic agent such as a phospholipid ln all these cases the polyanhydroglucuronic acid and salt thereof may contain in their polymeric chain from 8 to 30 percent by weight of carboxyl groups, at least 80 percent by weight of these groups being uronic groups, at most 5 percent by weight of carbonyl groups, and at most 0.5 percent by weight of bound nitrogen.

Preferably the polyanhydroglucuronic acid and salt thereof may contain in their polymeric chain at most 0.2 percent by weight of bound nitrogen.

The molecular mass of the polymeric chain of the polyanhydroglucuronic acid and salt thereof may be from about 1 X 103 to about 3 X 105 Daltons.

The molecular mass of the polymeric chain of the anionic component may range from about 5 X 103 to about 1.5 X 105 Daltons.

ALPE12/C The content of carboxyl groups may be in the range of from about 12 to about 26 percent by weight, at least 95 percent of these groups being uronic groups.

The polyanhydroglucuronic acid and salt thereof may contain at most 1 percent by weight of carbonyl groups.

Preferably, each carbonyl group may be an intra- or intermolecular 2,6 or 3,6 hemiacetal, a 2,4-hemialdal or a C2-C3 aldehyde.

The biocompatible intermolecular polymer complex may be a complex of: an anionic component comprising polyanhydroglucuronic acid or salt, which is that of a partially or completely hydrolysed and oxidative-environment hydrolysed polyanhydroglucuronic acid containing material; and a non protein cationic component comprising a linear or branched natural, semi-synthetic or synthetic oligomer or polymer.

Preferably, at least 5% of the basic structural units of the anionic component may be glucuronic acid.

The cationic component may contain nitrogen that either carries a positive charge or wherein a positive charge is induced by contact with the polysaccharidic anionic component.

The cationic component may be a member selected from the group consisting of a derivative of acrylamide, a derivative of methacrylamide, a copolymer of acrylamide and a copolymer of methacrylamide such as polyacrylamide, a copolymer of hydroxyethylmethacrylate and hydroxypropylmethacrylamide, and a copolymer of -14AI.PF.I2/C acrylamide, butylacrylate, maleicanhydride and methylmethacrylate. Preferably the cationic component may be a cationised natural polysaccharide.

The polysaccharide may be a starch, cellulose or gum. Preferably the gum may be guargumhydroxypropyltriammonium chloride.

The cationic component may be a synthetic or semi-synthetic polyamino acid. For example the cationic component may be a member selected from the group consisting of polylysin, polyarginin and .alpha, beta.-poly-[N-(2-hydroxyethyl)-DL-aspartamide]. Preferably the cationic component may be a synthetic anti-fibrinolytic such as hexadimethrindibromide.

The cationic component may be a natural or semi-synthetic peptide for example, the peptide may be a member selected from the group consisting of a protamine, gelatine, fibrinopeptide, and a derivative of one of the foregoing.

The cationic component may be an aminoglucane or derivative thereof such as a fractionated chitin or its de-acetylated derivative chitosan. Alternatively, the aminoglucane may be of microbial origin or is isolated from an arthropod shell.

The invention also provides a method for reducing the development of atherosclerosis in a mammal comprising the step of orally administering a therapeutic preparation comprising a biocompatible anionic polyanhydroglucuronic acid (PAGA) or salt thereof, said PAGA prepared by partial or complete hydrolysis and neutralisation of a PAGA containing material wherein the PAGA interacts with biomolecules in the fluid media of the gastrointestinal tract.

The invention further provides a method for treating the inflammation related to atherosclerosis comprising the step of orally administering a therapeutic preparation comprising a biocompatible anionic polyanhydroglucuronic acid (PAGA) or salt thereof.

A1.PEI2/C said PAGA prepared by partial or complete hydrolysis and neutralisation of a PAGA containing material wherein the PAGA interacts with biomolecules in the fluid media of the gastrointestinal tract.

The invention also provides a method for improving the management of blood glucose and insulin for patients with type 2 diabetes meliitus in a mammal, comprising the step of orally administering a therapeutic preparation comprising a biocompatible anionic polyanhydroglucuronic acid (PAGA) or salt thereof, said PAGA prepared by partial or complete hydrolysis and neutralisation of a PAGA containing material wherein the PAGA interacts with biomolecules in the fluid media of the gastrointestinal tract.

The invention further provides a method for treating type 2 diabetes associated cardiovascular problems comprising the step of orally administering a therapeutic preparation comprising a biocompatible anionic polyanhydroglucuronic acid (PAGA) or salt thereof, said PAGA prepared by partial or complete hydrolysis and neutralisation of a PAGA containing material wherein the PAGA interacts with biomolecules in the fluid media of the gastrointestinal tract.

The invention also provides a combination therapy for the benefit of cardiovascular health in a mammal comprising the step of orally administering a therapeutic preparation comprising of a cardiovascular health benefiting agent and a biocompatible anionic polyanhydroglucuronic acid (PAGA) or salt thereof, said PAGA prepared by partial or complete hydrolysis and neutralisation of a PAGA containing material wherein the PAGA interacts with biomolecules in the fluid media of the gastrointestinal tract.

Definitions The invention in particular involves the use of polyanhydroglucuronic acids, salts and intermolecular complexes (1MC) thereof. The term polyanhydroglucuronic acid, salts and IMCs thereof as used herein also includes copolymers thereof, especially with anhydroglucose. These as a whole are hereinafter referred to as PAGA. Throughout the AEPEI2/C specification PAGA is also referred to as microdispersed oxidised cellulose (MDOC) available from Alltracel Pharma Limited. MDOC is a registered trademark.

CS 242920, CS 292723, GB 2314840, but notably WO98/33822 describe particular polyanhydroglucuronic acids and salts thereof and a method of preparing such compounds. In particular the term polyanhydroglucuronic acids and salts thereof includes the acids and salts referred to in WO98/33822, the entire contents of which are incorporated herein by reference.

The partial or complete hydrolysis and neutralisation of the PAGA containing material is carried out in aqueous or water based organic solutions of inorganic or organic salts and bases and/or an oxidative environment. In this way the secondary products such as aldehydes and ketones and their condensation products inevitably produced as a result of the initial oxidation step are removed. These aldehyde and ketone impurities have a fundamental influence on the stability of the polyanhydroglucuronic acid (PAGA) product. A stable PAGA product with a reduced degree of crystallinity and a high degree of purity in a microdispersed form is produced.

WO00/05269 describes particular intermolecular complexes (lMCs) of PAGA. JMCs of PAGA in particular include those referred to in WO00/05269, the entire contents of which is incorporated herein by reference.

The term “non-protein” is intended to distinguish from what the applicants understand the commonly used definition in biochemistry of “protein” to be. According to IUPAC a protein is a naturally occurring polypeptide of specific sequence of genetically code aminoacids with more than about 50 residues and molecular weights over about 10,000 displaying supermolecular, so-called secondary, tertiary, and quaternary structure (cf. IUPAC Recommendations 1983: Nomenclature and Symbolism for Amino Acids and Peptides). Proteins differ from polypeptides in having higher molecular weights (by convention over 10,000) and more complex structure. -17ALPEI2/C The cationic components of the complexes described in the invention include inter alia small molecular weight peptides and aminoglycans. These components would be commonly understood to have a low molecular weight typically less than 5,000. For example the gelatine used in the invention is suitably hydrolysed which results in a reduction of the molecular weight. The gelatine used in the invention is described as a peptide which a person skilled in the art would take to infer a compound of low molecular weight.

However, other proteins, peptides or aminoglycans of significantly higher molecular weight may also be used to prepare PAGA IMCs as nutraceutics reducing glucose and cholesterol levels in plasma.

The AIN-93 synthetic diet (Journal of Nutrition vl23, 1943-44 (1993)) consists essentially of the ingredients listed in Table I below.

Table 1. ΑΓΝ-93 ingredients Ingredient mg’s/gram Casein 140.00 Cornstarch 465.692 Dyetrose 155.00 Sucrose 100.00 Cellulose 50.00 Soybean Oil 40.00 t-Butylhydroquinone 0.008 Salt Mix 35.00 Vitamin Mix 10.00 L-Cystine 1.80 Choline Bitartrate 2.50 -18ALPEI2/C Brief Description of the Figures The invention will be more clearly understood from the following description thereof with reference to the accompanying Figures, in which: Fig. I is a bar chart showing the total serum cholesterol levels, VLDL, LDL, HDL and TAG in a control group, a simvistatin group, an atorvastatin group and an oxidized cellulose (MDOC) group; Fig. 2 shows the immunohistochemical staining of endothelial expression of an inflammatory marker ICAM-1 in (A) a control group, (B) a simvistatin group, (C) an atorvastatin group and (D) an oxidized cellulose (MDOC) group. There is strong endothelial expression in control animals (a) and weaker expression in simvastatin (b), atorvastatin (c) and MDOC (d) treated animals. The 1CAM-I expression in atorvastatin treated animals (c) is visible in only a few endothelial cells (arrows). 200 x original magnification.; Fig. 3 shows the immunohistochemical staining of endothelial expression of an inflammatory marker VCAM-1 in A) a control group, (B) a simvistatin group, (C) an atorvastatin group and (D) an oxidized cellulose (MDOC) group. There is strong endothelial expression in control animals (a) and decreased expression in simvastatin (b), atorvastatin (c) and MDOC (d) treated animals. Very weak endothelial expression of VCAM-1 (arrows) was detected in the atorvastatin (c) and MDOC group (d). 200 x original magnification; Fig. 4 is a bar chart showing the percentage of activated endothelial cells in both the aortic root and the aortic arch. The expression of the inflammatory marker VCAM-1 is significantly decreased in atorvastatin and MDOC treated mice in + comparison to the control group *P < 0.001 versus the control group. P < 0.001 versus the control group. Moreover, atorvastatin significantly reduced expression -19li l\0 A1.PE12/C of the inflammatory marker ICAM-I in comparison to the control group. P < 0.001 versus the control group.

Fig 5 is a bar chart showing the percentage of activated endothelial cells in control, atorvastatin and MDOC groups.

Fig. 6 is a line graph showing weight gain during the course of a combination therapy study for control groups and treatment groups; Fig. 7 is a bar chart showing the effect of dietary supplementation in a combination treatment study of a high cholesterol diet (HCD) with 1% oxidised cellulose (OC) MDOC and 0.5% sterol (w/w) on the serum LDL cholesterol concentrations in rats (n=20/gp) fed these diets; Fig. 8 is a bar chart showing the effect of dietary supplementation in a combination treatment study of a high cholesterol diet (HCD) with 1% oxidised cellulose (OC) MDOC and 0.5% sterol (w/w) on the serum total cholesterol concentrations in rats (n=20/gp) fed these diets; Fig. 9 is a line graph showing weight gain during the course of a study on the effect of dietary supplementation on serum glucose concentrations for control and treatment groups; and Fig. 10 is a bar chart showing the effect of the dietary supplementation of HCD with 1 % and 5% MDOC (w/w) on the serum glucose concentrations in rats (n=10/gp) fed these diets.. p<0.05,»» p<0.01 and *»* p<0.001.

Detailed Description -20AEPEI2/C We have found an important use of PAGA a salt thereof, a copolymer thereof, or an intermolecular complex thereof in the preparation of a medicament for the treatment of inflammation and for the management of blood glucose levels. We have also found an important composition comprising PAGA, a salt thereof, a copolymer thereof, and an intermolecular complex thereof as a first component and a second component which is an antilipemic agent.

Preferably the oxidised polysaccharide material is in the form of biocompatible anionic polyanhydroglucuronic acid (PAGA), salt or intermolecular complex (1MC) thereof. The PAGA may be prepared by partial or complete hydrolysis and neutralization in solutions of (in)organic hydroxides, salts or bases in a standard (CZ242920) aqueous and/or aqueous/organic, or an oxidative environment (WO98/33822). During this process salts are formed, for example salts with Na ions, or complex salts with Ca, Al, Fe ions but also with aminoacids of lysin, arginin, or histidin types. IMCs are also formed for example with gelatin or peptides of hydrolysed collagen, but also with blood proteins or aminoglycans as described in WO00/05269. The salts and IMCs formed are dependent on the hydrolysis conditions and types of salt, base and /or the mixtures used.

In a particularly preferred embodiment the polysaccharide material is polyanhydroglucuronic acid, biocompatible salts thereof, copolymers thereof or a biocompatible intermolecular complex polymer thereof. Preferably the oxidised polysaccharide is derived from cellulose, starch, or gum, or is of microbial origin.

In contrast to soluble or insoluble dietary fibres, the PAGA, its salts or IMCs, are prepared by hydrolysis from a raw oxidised polysaccharide, well defined, from a pharmacological point of view, by IR, NMR, GPC spectra, elemental analysis, and cation content.

Similar to hyaluronic acid (HA), the PAGA displays reducing ability in a biological environment, yet with a higher assimilable organic carbon (AOC) value. -21ALPEI2/C As such, PAGA and derivatives thereof prepared by the above methods is capable of forming a highly hydrated film on a biological surface such as gastrointestinal tract (GIT) mucous tissue.

Despite the fact that hydrolytically prepared PAGA yields low viscous water solutions, its IMCs with for example collagen, chitosan or other polymeric cations have a high viscosity. Such IMCs may be created as products during the hydrolysis or afterwards in situ - on the mucous tissue of the gastrointestinal tract - out of PAGA being administered and peptides or proteins involved in food. They also display a higher osmolality than simple PAGA salts (such as Na salt).

In clinical trials with volunteers it has been observed that oral administration of PAGA reduced LDL and total cholesterol and had the effect of increasing the HDL cholesterol level. In these clinical trials it was surprisingly found that PAGA in the form of a Ca/Na salt had an acute transitory effect of favourably influencing the lipid spectrum in including the ratios (LDL/HDL)/TG and TC/TG by short-term oral administration to humans.

In addition the behaviour of the atherogenic index of plasma (AIP), (Dobiasova M., Clinical Chemistry. 2004;50:l 113-1115) was affected. AIP is expressed as the logarithm of the ratio of molar concentrations (triglycerides/HDL-cholesterol) and strongly correlates with the size of LDL-C particles and the cholesterol esterification rate in plasma deprived of apoB lipoproteins. These indices are currently considered as the most sensitive indicators of the plasma atherogeny profile. The parameter proved highly significant in terms of absolute differences and was significantly reduced after the first phase of PAGA administration.

A further surprising effect was the markedly favourable effect of PAGA in contrast to lactose used as placebo on glycaemia and HbAlc values. The clearly documented -22ALPF.I2/C reduction of glycaemic response may lead to reduction of insulin production and thereby to a decrease in hepatic cholesterol synthesis.

A reduction of bilirubin values in probands of the PAGA group was also noticeable which may indicate an increased offer of substrate for enzymatic transformation (glucuronation) in liver and involvement in metabolic pathways of lipid management. These results were however statistically insignificant.

The results clearly indicate the potential favourable effect of PAGA and derivatives thereof on the values of lipid and saccharide management in an organism. The therapeutic benefit is significant for an efficient hypolipidemic, or as the case may be an antidiabetic, in persons with metabolic syndrome (the occurrence of which is assessed to be up to 20 to 30 % of both the European and American populations) and in Type 2 diabetics (on average 7 % of registered ones and about 15 % of latent ones in our population.

Similar cholesterol management results were found in pre-clinical testing.

Biochemical analysis showed PAGA treatment resulted in a very mild and insignificant lowering of total serum cholesterol in mice in comparison to the control group. Endothelial expression of VCAM-1 in the aorta significantly decreased in PAGA (MDOC) treated mice when compared to non-treated mice. We found the potential of the new hypolipidemic substance PAGA to decrease endothelial expression of VCAM-1 in very early stages of atherogenesis in apoE-deficient mouse model of atherosclerosis.

This potential anti-inflammatory effect may be related to the mild hypolipidemic effect of PAGA.

It may be hypothesized that the mechanism of this effect is related to interactions with biomolecules in the fluid media of the gastrointestinal tract which have been demonstrated by in vitro surface plasmon resonance scientific investigation. -23ALPE12/C An advantage of the therapy ofthe invention is the inherent biocompatibility, lack of toxicity and virtual absence of adverse side effects inherent to PAGA salts and intermolecular complexes (IMC) thereof. This reduces the potential risks to the subject compared with other types of antilipemic remedies.

Another advantage is the fact that in addition to its own therapeutic effects it has synergistic properties for use in combination with antilipemic compositions such as notably statins, sterols, stanols, CETP inhibitors, dietary fibre, fibrates and niacin.

The composition of the present invention may be used as an effective agent to lower serum cholesterol in animals, particularly humans. It should be understood, however, that this composition is equally suited for administration to other animals, for example, in the form of veterinary medicines and animal foods.

In another form of the present invention, the composition of the present invention may be incorporated into foods, beverages and nutraceuticals, including, without limitation, the following: 1) Dairy Products--such as cheeses, butter, milk and other dairy beverages, spreads and dairy mixes, ice cream and yoghurt; 2) Fat-Based Products-such as margarines, spreads, mayonnaise, shortenings and dressings; 3) Cereal-Based Products—comprising grains (for example, bread and pastas) however processed; 4) Confectionaries—such as chocolate, candies, chewing gum, desserts, non-dairy toppings, sorbets, icings and other fillings; -24ALPEI2/C ) Beverages—whether alcoholic or non-alcoholic and including colas and other soft drinks, juices, water, dietary supplement and meal replacement drinks such as those sold under the trade-marks Boost™, and Ensure™; and 6) Miscellaneous Products-including eggs, processed foods such as soups, pre-prepared pasta sauces, pre-formed meals and the like.

The composition of the present invention may be incorporated directly and without further modification into the food, nutraceutical or beverage by techniques such as mixing, infusion, injection, blending, immersion, spraying and kneading. Alternatively, the composition may be applied directly onto a food or into a beverage by the consumer prior to ingestion.

It is contemplated within the scope of the present invention that the composition of the present invention may also be incorporated into various conventional pharmaceutical preparations and dosage forms such as tablets (plain and coated) for use orally, capsules (hard and soft, with or without additional coatings) powders, granules (including effervescent granules), pellets, microparticulates, solutions (such as micellar, syrups, elixirs and drops), lozenges, pastilles, ampuls, emulsions, microemulsions, suppositories, gels, modified release dosage forms together with customary excipients and/or diluents and stabilizers.

Examples of an Excipient preparation Excipient 1 “hydrophilic” (Fluka) Macrogolum 1500..............50% w/w Macrogolum 400..............40% PAGA Na salt................... 2% D,L-a-Tocopherol acetate..... 2% D-Panthenol......................4% -25It** AI.PEI2/C Excipient 2 Ceresine (white wax) 65/70.................2.34 % w/w Parafine 52/40.................................2.64 Beef fat.......................................17.00 Vaseline oil..................................30.94 Vaselinum album...........................20.00 PAGA Ca/Na salt............................2.00 (micro and/or nanoparticles) Zinc stearate..................................5.08 Emulgator ©.................................20.00 Preparation Examples Example A - Preparation of Polymer Complexes Materials: long-fibre cotton-medicinal cotton wool oxidised by NxOy (proprietary); C6OOH 18.8% b/w; ash content <0.1 % b/w; ZC=O 0.6% b/w % solution Na2CO3 (Lachema, a.s. Neratovice) CaC12.6H2O anal.grade (Lachema, a.s. Neratovice) demineralised water 2 .mu.S, ethanol, synthetic rectified cone. 98% (Chemopetrol Litvinov, a.s.) acid acetic anal.grade (Lachema, a.s. Neratovice) H2O2 anal.grade 30% (Lachema, a.s. Neratovice) N-HANCE 3000 guargumhydroxypropyltriammoniumchloride (Aqualon—Hercules) ALPE12/C Equipment: mixer: bottom stirring, 1501 (duplicator), stainless steel EXTRA S vibrating screen: stainless steel, 150 mesh rotary air pump: rotor diameter 150 mm turbostirrer: ULTRA TURAX (Janke-Kunkel) beaker: 51 pH meter PICCOLO thermocouple thermometer Procedure: g of N-HANCE 3000 were placed into a 5 I beaker and 3 1 of demineralised water 2 .mu.S were added. Contents of the beaker were intensely stirred for 30 minutes. The pH value was adjusted to less than 4.5 by addition of an acetic acid solution leading to a viscosity rise. 60 1 of demineralised water 2 .mu.S were introduced into a mixer. Then 3 kg of CaCI2.6H2O anal.grade were added and the contents heated up to a temperature of 50°C. under stirring. On dissolution of the calcium chloride the stirring was interrupted and 2.7 kg of the raw oxidised cotton wool were introduced. The mixer was closed and the contents were agitated for 120 seconds. Then the pH value of the contents was adjusted by addition of a 20% solution of Na2CO3 to 6-6.5 and 13 kg of H2O2 30% were introduced. The fibre suspension was slowly agitated for 10 minutes. Then the pH value was readjusted to 4.5 to 5.0 and the prepared viscous solution of N-HANCE 3000 was introduced. The contents of the mixer were stirred intensely for 30 seconds. Subsequently 60 1 of synthetic rectified ethanol cone. 98% were introduced into the mixer. After another 15 seconds from adding the ethanol the contents of the mixer were transferred onto a vibrating screen, and the supernatant. Liquid was filtered off. The filtration cake was redispersed in the mixer in 60 1 of a mixture of 18 I of synthetic rectified ethanol cone. 98% and 42 1 of demineralised water 2 .mu.S. The fibre suspension was filtered again on the vibrating screen.

AEPEI2/C The isolated material thus prepared may further serve to prepare final products of the nonwoven type via a wet or dry process.

Analysis Ca content 4.0% b/w Na content 1.8% b/w ZC=O content 0.0% b/w COOH content 20.7% b/w Example B - Preparation of Polymer Complexes (1MC-MDOC) Materials: oxidised short-fibre cotton (Linters -Temming) (proprietary) C6OOH 16.8% b/w ash content <0.15% b/w LC=O 2.6% b/w NaOH anal.grade (Lachema, a.s. Neratovice); redistilled water (PhBs 1997); ethanol, synthetic rectified cone. 98% (Chemopetrol Litvinov, a.s.); isopropanol 99.9% (Neuberg Bretang); H2O2 anal.grade 30% (Lachema, a.s. Neratovice); gelatine (PhBs 1997) Equipment: turbostirrer: ULTRA TURAX (Janke-Kunkel) sulphonation flask 1 1 heater 1.5 kW laboratory centrifuge: 4000 rpm thermostated water bath pH meter PICCOLO glass thermometer rotary vacuum dryer or hot-air dryer ALPE12/C Procedure: Into a 1 1 sulphonation flask equipped with a turbostirrer and a heater, 400 ml of redistilled H2O were placed, and 8 g of NaOH were added. On dissolution, 50 g of oxidised Linters were added, the contents were heated up to 70°C and the stirring intensity set to a maximum. After 20 minutes, 40 g of 30% H2O2 were added into the flask, temperature was increased to 85°C, and maintained for another 10 minutes. The contents were then cooled down to 50°C on a water bath and gelatine solution (10 g of gelatine in 70 g of redistilled H20) warmed up to 50°C was added to the hydrolysate.

The temperature was decreased to 25-30°C and the pH of the system was checked and adjusted to a value of 6.0-6.5. Subsequently, 626 ml of synthetic rectified ethanol cone. 98% were added gradually under intense stirring. The suspension of IMC thus formed was isolated using a laboratory centrifuge. The supernatant liquid was filtered away and the cake was redispersed into 250 ml of 50% ethanol. The system was centrifuged again and after the separation of the supernatant liquid, the IMC was redispersed into 250 ml of synthetic rectified ethanol cone. 98% and let to stay for 4 hours. It was then centrifuged again, redispersed into 99.9% isopropanol, and let to stay for a minimum of 10 hours at 20°C. The gel formed was centrifuged again and the product was dried in a rotary vacuum dryer or a hot-air dryer. The product can be used, for instance, for microembolisation, for preparation of haemostatic dusting powders, for manufacture of polymer drugs, e.g. based on cytostatics, or for preparation of spheric particles for macroembolisation.

Analysis Na content 3.8% b/w LC=O content 0.0% b/w COOH content 21.5% b/w N content 2.7% b/w -29AL PE 12/C Example C - Preparation of Tablets and Pellets from MDOC (Microdispersed Oxidised Cellulose) Materials: MDOC (Ca/Na salt of PAGA), particle size 0.1-2.0 .mu.m, specific surface area 86 m2/g, COOH group content 22.2% b/w, Ca content 4.2% b/w, Na content 3.8% b/w Equipment: tabletting machine (KORSCH EK 0, Berlin) Procedure: 100 g of MDOC were introduced into the tabletting machine. The tabletting force was set at a value of 5 kN.

The tablets prepared were smooth and cohesive and had a weight of 0.5 g. Disintegration rate of the tablets in a saline FI/1 was 15 minutes at 20°C, and 8 minutes at 37°C.

Example D A patient aged 55, displaying an increased cholesterol content in blood was treated by MDOC tablets administered orally for 50 days, at a dose of 6 tablets daily. After the treatment both LDL content and total cholesterol content were significantly reduced.

Blood analysis: Total Cholesterol HDL LDL Triacylglycerols before treatment 8.60 mmol/1 1.16 mmol/1 6.50 mmol/1 2.70 mmol/1 after treatment 6.60 mmol/1 1.20 mmol/1 4.90 mmol/1 2.40 mmol/1 -30AI.PEI2/C Example E - Preparation of Tablets and Pellets from 1MC-MDOC Complex Preparation of IMC-MDOC complex Material: oxidised short-fibre cotton (Linters-Temming) (proprietary) C6OOH 16.8% b/w ash content < 0.15 % b/w Σ C=O 2.6 % b/w % solution Na2CO3 (Lachema, a.s. Neratovice) CaCI2.6H2O anal.grade (Lachema, a.s. Neratovice) redistilled water (PhBs 1997) ethanol, synthetic rectified cone. 98% (Chemopetrol Litvinov, a.s.) isopropanol 99.9% (Neuberg Bretang) H2O2 anal.grade 30% (Lachema, a.s. Neratovice) gelatine (PhBs 1997) Equipment: turbostirrer: ULTRA TURAX (Janke-Kunkel) sulphonation flask 1 litre heater 1.5 kW laboratory centrifuge: 4000 rpm thermostated water bath pH meter PICCOLO glass thermometer rotary vacuum dryer or hot-air dryer Procedure: Into a 1 litre sulphonation flask equipped with a turbostirrer and a heater, 400 ml of redistilled H2O were placed, 15.73 g of CaC12.6H2O were added and on dissolution, 40.0 g of 20%Na2CO3 solution were introduced under stirring. Subsequently, 50 g of oxidised Linters were added to the white emulsion formed and the contents were heated up to 95°C with the stirring intensity set to a maximum. After 10 minutes, 30 g of 30% H2O2 were added into the flask and the hydrolysis continued for another 10 minutes.

ALPFI2/C The contents were then cooled down to 60°C on a water bath and the pH of the system was adjusted to a value of 4.5 - 5.0 by the addition of a 20% solution of Na2CO3. A gelatine solution (10 g of gelatine in 70 g of redistilled H2O) warmed to 50°C was added and left to react for another 20 minutes. The flask contents were then cooled to 30°C in a water bath and 626 ml of synthetic rectified ethanol cone. 98% were added gradually under intense stirring. The suspension of 1MC thus formed was isolated using a laboratory centrifuge. The supernatant liquid was filtered away and the cake was redispersed into 250 ml of 50% ethanol. The system was centrifuged again and after the separation of the supernatant liquid, the IMC was redispersed into 250 ml of synthetic rectified ethanol cone. 98% and alllowed stand for 4 hours. It was then centrifuged again, redispersed into 99.9 % isopropanol, and left to stand for a minimum of 10 hours at 20°C. The gel formed was centrifuged and the product was dried in a rotary vacuum dryer or a hot-air dryer.

The product can be used, for instance, for microembolisation, for preparation of haemostatic dusting powders, for manufacture of polymer drugs, e.g. based on cytostatics, or for preparation of spheric particles for macroembolisation.

Analysis: content Ca 4.4 % b/w content Na 2.7 % b/w content Σ C=O 0.0 % b/w content COOH 20.5 % b/w content N 1.8 % b/w Preparation of Tablets and Pellets from 1MC-MDOC Complex magnesium stearate (SIGMA) ascorbic acid (MERCK) α-tocoferol acetate (Slovakofarma Hlohovec) -32ALPEI2/C ethanol synthetic rectified (Chemopetrol Litvinov, a.s.) Equipment; tabletting machine (KORSCH EK 0, Berlin) blender (Nautamix 300) counter-flow drier (BINDER) Procedure; kg of IMC-MDOC complex of composition according to Example 2 were placed into the blender. 660 g of micronised ascorbic acid, 1660 g of α-tocoferol acetate emulgated in 2500 ml of ethanol and 1000 g of magnesium stearate were added. The mixture was homogenised for 3 hours and dried in a counter-flow drier at a temperature of 50°C until the ethanol was removed. 100 g of the resulting dry powder were introduced into the tabletting machine. The tabletting force was set at a value of 7 kN.

The tablets prepared were smooth and cohesive and had a weight of 0.5 g. Disintegration rate of the tablets in a saline F1/1 was 17 minutes at 20°C, and 8 minutes at 37°C.

Example F A patient aged 57, displaying an increased cholesterol content in blood was treated by IMC-MDOC tablets administered orally for 50 days, at a dose of 6 tablets daily. After the treatment both LDL content and total cholesterol content were significantly reduced.

Blood analysis before treatment after treatment Total Cholesterol 7.70 mmol/1 5.70 mmol/1 HDL 1.16 mmol/1 1.30 mmol/1 LDL 4.40 mmol/1 3.30 mmol/1 Triacylglycerols 1.81 mmol/1 1.80 mmol/1 -33ALPE12/C Example G - Preparation of Granules from MDOC in a Fluid Bed Materials: MDOC, particle size 0.1-2.0 .mu.m, specific surface area 86 m2/g, COOH group content 22.2% b/w, Ca content 4.2% b/w, Na content 3.8% b/w Equipment: Set of vibrating screens with mesh size 100, 150, 200, 250, 350, 500 .mu.m mixer, bottom agitated, vessel size 1000 ml, 8000 rpm, equipped with a nozzle for inlet of the granulation medium counter-flow drier (BINDER) Procedure: 100 g of MDOC were placed into the mixer, the mixer was closed and the agitation begun. A water mist was gradually injected into the mixer at a rate of 10 g/45 seconds. The granulate formed was transferred to the counter-flow drier and dried at a temperature of 45 °C until the humidity content was reduced to below 6% b/w. The dried granules were sieve-screened using the set of vibrating screens. The individual fractions were packaged into glass vials in amounts of 0.5-2.0 g each as required. The preparation was sterilised by y irradiation with a dose of 25 kGy.

The product may be used as a) an embolisation agent, or b) an antilipemicum.

Example H - Preparation of Granules from 1MC-MDOC Complex Materials: 1MC-MDOC complex as prepared in Example 2 Equipment: Set of vibrating screens with mesh size 100, 150, 200,250, 350, 500 .mu.m mixer, bottom agitated, vessel size 1000 ml, 8000 rpm, equipped with a nozzle for inlet of the granulation medium counter-flow drier BINDER.

Procedure: 100 g of MDOC were placed into the mixer, the mixer was closed and the agitation AEPE12/C begun. Saturated water vapour was gradually injected into the mixer at a rate of 10 g/45 seconds. The granulate formed was transferred to the counter-flow drier and dried at a temperature of 45°C until the humidity content was reduced to below 6% b/w. The dried granules were sieve-screened using the set of vibrating screens. The individual fractions were packaged into glass vials in amounts of 0.5-2.0 g each as required. The preparation was sterilised by gamma irradiation with a dose of 25 kGy.

The product may be used as a) an embolisation agent, or b) an antilipemicum.

The invention will be more clearly understood from the following Examples thereof given by way of illustration only.

EXAMPLES Materials and methods Animals The Ethical Committee of the Faculty of Pharmacy, Charles University, approved the protocols of the animal experiments. The protocol of experiments was pursued in accordance with the directive of the Ministry of Education of the Czech Republic (No. 311/1997).

ApoE ~ ~ mice ApoE' ' mice, generated by gene targeting, have been shown to develop pronounced hypercholesterolemia and atherosclerotic lesions (Reddick et al., 1994) with certain features resembling those seen in humans (Nakashima et al., 1994) and other species (Davies et al., 1988). ApoE7' mice exhibit spontaneous elevation of total plasma cholesterol and triglycerides and reduced levels of HDL on a diet with normal fat content and with no cholesterol supplementation (Zhang et al., 1992). Male apoE mice on a C57BL/ 6J background (n =32) weighing 10-15 g were kindly provided by Prof. Poledne (IKEM, Prague, Czech Republic) and housed in the SEMED, (Prague, Czech Republic).

ALPE12/C Example 1 - Serum Cholesterol Levels (a) Experimental design Male apol/' mice were weaned at 5 weeks of age and randomly subdivided into four groups. The control group of animals (n =8) was fed with the standard laboratory diet (chow diet) for another 4 weeks after the weaning. In both simvastatin (n =8) and atorvastatin (n =8) group, statins were added to the chow diet at the dosage of 10 mg/kg per day. In MDOC group (n =8) MDOC was added to the chow diet at the dosage 50 mg/kg per day. All treated mice were fed with the experimental diet for another 4 weeks after weaning with water ad libitum throughout the study. Each mouse, in both statins and MDOC group, lived in a separate cage obtaining 6 g of food (in specially prepared pellets) daily. The food consumption was monitored every day. No differences in the food consumption were visible, either among animals of one experimental group nor between experimental groups. The dose of simvastatin and atorvastatin used in the present study was based on the doses used in previous studies with hyperlipidemic mice (Laufs et al., 1998; Sparrow et al., 2001). The dose of MDOC was based on the results of a small pilot study. At the end of the treatment period, all animals were fasted overnight and euthanized. Blood samples were collected via cardiac puncture at the time of death. The aortas, attached to the top half of the heart, were removed and then immersed in OCT (Optimal Cutting Temperature) embedding medium (Leica, Prague, Czech Republic), snap frozen in liquid nitrogen cooled 2-methylbutane and stored at -80 °C. (b) Biochemistry Serum lipoprotein fractions were prepared using NaCI density gradient ultracentrifugation (Beckman TL 100. Palo Alto. CA). The lipoprotein fractions were distinguished in the following density ranges: very low density lipoprotein (VLDL)1.063 g/ml. Total concentration and lipoprotein fraction concentration of . 36 - “ ALPEI2/C cholesterol were assessed enzymatically by conventional diagnostic kits (Lachema. Brno, Czech Republic) and spectrophotometric analysis (cholesterol at 510 nm, triglycerides, at 540 nm wavelength), (ULTROSPECT Ill, Pharmacia LKB Biotechnology, Uppsala. Sweden). (c) Immunohistochemistry Sequential tissue sectioning started in the mouse heart until the aortic root containing semilunar valves together with the aorta appeared. From this point on, serial crosssections (7 Am) were cut on a cryostat and placed on gelatin-coated slides. Sections were air-dried and then slides were fixed for 20 min in acetone at -20 °C. Endogenous peroxidase activity was blocked with 0.3% hydrogen peroxide in phosphate buffered saline (PBS) for 15 min. After blocking of nonspecific binding sites with 10% normal horse serum (Sigma-Aldrich Chemie, Steinheim, Germany) in PBS solution (pH 7.4) for 30 min, slides were incubated with primary antibodies for 1 h at room temperature. After a PBS rinse, the slides were developed with biotinylated horse-anti goat IgG antibody or donkey antisheep IgG, in the presence of 200 mg/mL normal mouse IgG. Antibody reactivity was detected using HRPconjugated biotin- avidin complexes (Vector Laboratories, USA) and developed with diaminobenzidine tetrahydrochloride as substrate. Specificity of the immunostaining was assessed by staining with nonimmune isotype-matched immunoglobulins. Primary antibodies included the following: polyclonal goat anti-mouse 1CAM-I (M- 19, IgG) and polyclonal goat antimouse VCAM-1 (C-19, IgG) diluted 1:100 purchased from Santa Cruz Biotechnology (California. USA) and sheep antihuman Von Willebrand factor (PC054 IgG) purchased from The Binding Site (Birmingham. England), diluted 1: 300. (d) Quantitative analysis ofthe immunohistochemistry Stereological methods for the estimation of immunohistochemical staining of VCAM-1. 1CAM-1, and Von Willebrand factor were used as previously described (Nachtigal et al., 2002, 2004). In brief, the systematic uniform random sampling and the principle of the point-counting method were used for the estimation (Weibel, 1979). A total number of 50 -37ALPEI2/C consecutive serial cross-sections were cut into 7 f m thick slices, which gave us 0350 mm lengths of the vessel called the reference volume. This reference volume comprises several sections of the vessel containing semilunar valves in aortic root, and several sections of aortic arch (ascending part of the aorta). A systematic uniform random sampling was used in the reference volume. The first section for each immunohistochemical staining was randomly positioned in the reference volume and then each tenth section was used, thus five sections for each staining were used for the stereological estimation. The point-counting method was used and more than 200 test points per vessel, hitting immunostaining, were counted for an appropriate estimation (Gundersen et al., 1988). The estimated area is then; estA = a*P where the parameter: a characterizes the test grid; and P is the number of test points hitting either the atherosclerotic lesion or positive immunostaining.

The area of Von Willebrand factor expression was considered as a total area of intact endothelium. Thus, the area of VCAM-1. and ICAM-1 expression indicates the percentage of activated endothelial cells calculated as est P ~ area(x) * 100% area (VonWill) Where: area (x) is the area of VCAM-1 or ICAM-1 in the endothelium and area (Von Will) is the area of Von Willebrand factor expression in the endothelium. (e) Images -38AI.PEI2/C Photo documentation and image digitizing from the microscope were performed with the Nikon Eclipse E2000 microscope, with a digital firewire camera Pixelink PL-A642 (Vitana Corp. Ottawa, Canada) and with image analysis software LUCIA version 4.82 (Laboratory Imaging, Prague, Czech Republic). Stereological analysis was performed with a PointGrid module of the ELLIPSE software (ViDiTo, Kosice.

Slovakia). (f) Statistical analysis All values in the graphs are presented as a meanTSEM of n =8 animals. Statistical significance in the differences between groups was assessed by ANOVA followed by the Tukey test for multiple comparisons with the use of the SigmaStat software (version 3.0). P values of 0.05 or less were considered statistically significant. (g) Atherosclerosis Animals are to be killed by pentobarbital injection. Thoracic aortas are rapidly removed, immersion fixed in 10% neutral buffered formalin, and stained with oil red O (0.3%). After a single longitudinal incision along the wall opposite the arterial ostia, the vessels are pinned open for evaluation of the plaque area. The percent plaque coverage is determined from the values for the total area examined and the stained area, by threshold analysis using a true color image analyzer (Videometric 150; American lnnovision, Inc 1, San Diego, Calif.) interfaced to a color camera (Toshiba 3CCD) mounted on a dissecting microscope. Tissue cholesterol will be measured enzymatically as described, after extraction with a chloroform/methanol mixture (2:1) according to the method of Folch et al. (J. Biol. Chem., 226, 497-509 (1957)). (h) In Vitro Vascular Response The abdominal aortas are rapidly excised, after injection of sodium pentobarbital, and placed in oxygenated Krebs-bicarbonate buffer. After removal of perivascular tissue, 3mm ring segments are cut, placed in a 37°C muscle bath containing Krebs-bicarbonate solution, and suspended between two stainless steel wires, one of which is attached to a ALPF.I2/C force transducer (Grass Instrument Co., Quincy, Ma.). Force changes in response to angiotensin II added to the bath will be recorded on a chart recorder. (i) Results -/5 Hyperlipidemic apolipoprotein-E-deficient (apoE ) mice received normal chow diet or diet containing micro dispersed derivatives of oxidised cellulose (MDOC) 50 mg/kg/day or simvastain, atorvastatin lOmg/kg/day. Total cholesterol, VLDL, LDL, HDL and TAG were measured.

Determination of Serum Cholesterol (SER.CHOL, HDL-CHOL, TGI and VLDL+LDL) Total serum cholesterol (SER.CHOL) are to be measured enzymatically using a commercial kit from Wako Fine Chemicals (Richmond, Va.); Cholesterol Cl I, Catalog No. 276-64909. HDL cholesterol (HDL-CHOL) will be assayed using this same kit after precipitation of VLDL and LDL with Sigma Chemical Co. HDL Cholesterol reagent, Catalog No. 352-3 (dextran sulfate method). Total serum triglycerides (blanked) (TGI) will be assayed enzymatically with Sigma Chemical Co. GPO-Trinder, Catalog No. 337B. VLDL and LDL (VLDL+LDL) cholesterol concentrations will be calculated as the difference between total and HDL cholesterol.

Biochemical analysis showed that statins treatment did not decrease levels of total cholesterol and VLDL (Fig. 1). By contrast, atorvastatin treatment increased levels of the total serum cholesterol in comparison to the control group (23.65 ± 2.09 vs. 21.62 ± 2.94 mmol/1). However, MDOC treatment decreased total serum cholesterol in comparison to the control group (17.35 ± 2.64 vs. 21.62 ± 2.94 mmol/1) (Fig. 1).

Example 2 -Endothelial expression of VCAM-1 in apoE’7’ mouse model of atherosclerosis -40AI.PE12/C Members of the immunoglobulin superfamily of endothelial adhesion molecules, vascular cell adhesion molecule (VCAM-1) and intercellular cell adhesion molecule (1CAM-1), strongly participate in leukocyte adhesion to the endothelium and play an important role -/in all stages of atherogenesis. We used an apoE mice atherosclerotic model to detect and quantify the changes of endothelial expression of VCAM-1, and ICAM-1 in the vessel wall after the short-term administration of MDOC, simvastatin and atorvastsatin.

Hyperlipidemic apolipoprotein-E-deficient (apoE ) mice received normal chow diet or diet containing micro dispersed derivatives of oxidised cellulose (MDOC) 50 mg/kg/day or simvastain, atorvastatin lOmg/kg/day. Total cholesterol, VLDL, LDL, HDL and TAG were measured and the endothelial expression of VCAM-1 and ICAM-1 was visualized and quantified by means of immunohistochemistry and stereology, respectively.

Immunohistochemical staining of VCAM-1 and ICAM-1.

No atherosclerotic lesion or other morphological abnormalities in the aortic arch were visible in any mice in the experiment. Von Willebrand factor expression was observed only in endothelial cells in all groups of mice and this antibody was used as standard for the detection of intact endothelium (data not shown). The expression of VCAM-1 and ICAM-1 was observed in vessel endothelium in all groups of animals (Fig. 2, 3). Furthermore, ICAM-1 expression was stronger then VCAM-1 in each experimental group. Moreover, ICAM-1 expression decreased in both statins (Fig. 2b, c), and MDOC (Fig. 2d) treated mice compared to the control group (Fig. 2a). The same results were observed even for VCAM-1 staining (Fig. 3). However, the strongest diminution of ICAM-1 and VCAM-1 expression was visible in the atorvastatin treated mice (Fig 2c, 3c).

The expression of ICAM-1 and VCAM-1 in endothelium was related to the von Willebrand staining of the endothelium, thus the results indicate the percentage of activated endothelial cells. Results of the stereological analysis confirmed that ICAM-1 staining was much stronger in all mice compared to the VCAM-1 staining (Fig. 4). The AI.PE12/C percentage of activated endothelial cells ICAM-l/von Willebrand slightly decreased in simvastatin (22.38 ± 2.99 vs. 28.71 ±5.12 %) and MDOC (20.32 ± 3.40 vs. 28.71 ±5.12 %) treated animals in comparison to the control mice. However, significant diminution of both ICAM-l/von Willebrand (2.97 ± 1.42 vs. 28.71 ±5.12 %, P < 0.001) and VCAM1/von Willebrand (2.11 ± 0.98 vs. 14.82 ± 1.83 %, P < 0.001) staining was observed in atorvastatin treated animals in comparison to the control group (Fig. 4). In addition, VCAM-l/von Willebrand staining significantly decreased even in MDOC treated animals (3.35 ± 1.80 vs. 14.82 ± 1.83 %, P < 0.001) (Fig. 4).

Example 3 - VCAM expression in activated endothelial cells (a) Animals Male apoE'' mice on a C57BL/6J background (n=24) were weaned at 5 weeks of age and randomly divided into 3 groups.

Control group (n=8) - mice consumed a standard diet for 4 weeks (6 g of diet per day).

MDOC group (n=8) - mice consumed a standard diet supplemented with MDOC (50 mg/kg per day) for 4 weeks (6 g of diet per day).

Atorvastatin group (n=8) - mice consumed a standard diet supplemented with atorvastatin (10 mg/kg per day) for 4 weeks (6 g of diet per day).

No differences in the average weekly food consumption and body weight were detected during the study. At the end of the treatment all animals were fasted overnight, and euthanized. Blood samples were collected at the time of death via cardiac puncture. (b) Quantitative analysis of immunohistochemistry Sequential 20 pm sections of the aortic lumen were cut, dried and fixed. Estimation of immunohistochemical staining of VCAM-1 and ICAM-1 was carried out. In brief, -42AI.PKI2/C systematic uniform random sampling and the principle of point cutting were used for this determination. Fifty consecutive serial cross sections were cut in to 7 pm thick slices. Each tenth section was used, thus five sections for each staining were used for the stereological estimation. The area of von Willebrand factor expression was considered as a total area as the total area of intact endothelium. Thus, the area of VCAM and ICAM expression is defined as the percentage of the von Willebrand factor expression (i.e. the total endothelium). This method is described in more detail in Nachtigal et al (2004).

The percentage of VCAM expressing activated endothelial cells was calculated as described in the study design section. The results for the control, atorvastatin and MDOC groups were compared and the means were plotted in Figure 5.

Table 2 - Statistical results for VCAM expression in the control, atorvastatin and MDOC groups.

Treatment Control Atorvastatin MDOC Mean 14.82 2.11 3.35 SEM 1.83 0.98 1.8 Size 8 8 8 Tukey Test (p) <0.001 <0.001 The results presented in Figure 5 demonstrate that atorvastatin treatment greatly reduced the expression of VCAM (p<0.001). The MDOC group also had a significantly reduced VCAM expression (p<0.001).

These pre-clinical studies have demonstrated that MDOC is able to significantly decrease (77%) the expression of VCAM-1 in the aorta. VCAM is intimately linked to the development of atherosclerotic plaques and therefore its reduction by MDOC may decrease the chance of atherosclerosis. This conclusion adds further valuable evidence for a significant role in cardiovascular disease reduction by MDOC. -43ALPEI2/C The mechanism by which this effect occurs is most likely due to the dietary reduction of LDL by MDOC. Atherosclerotic arteries produce excess reactive oxygen species such as super oxide anion O2 , promoting oxidative modification of LDL. Oxidised LDL (oxLDL) accumulates in the atheroma and promotes macrophage survival and growth and monocyte proliferation and hence foam cell development. The accumulation of such inflammatory molecules in the atheroma leads to production of various proteinases, reactive oxygen species and contributes to endothelial dysfunction, plaque vulnerability and thrombogenicity. The lowering of LDL by MDOC may therefore reduce the amount of oxLDL produced. Hence, the reduced accumulation of oxLDL might contribute to decreased numbers of macrophages in atheroma during lipid lowering and hence the atherosclerotic risk.

This shows the potential of the hypolipidemic substance MDOC to decrease endothelial expression of VCAM-1 in very early stages of atherogenesis. This potential antiinflammatory effect may be related to the mild hypolipidemic effect of MDOC.

Example 4 - Combination therapy (a) Animals Rats with a body weight of approximately 175g were obtained from Charles River Laboratories, Ballina, Ireland. After 1 wk of acclimation, 80 rats were allocated to 4 similar groups (w = 20 per group) on the basis of their body weight. The rats were housed in groups of two and three in standard cages with autoclaved dust-free processed sawdust as bedding material. The rats were housed in an environmentally controlled room (temperature 22 ± 3°C) with a 12-h light:dark cycle. Throughout the study, the rats had free access to food and drinking water. Clinical observation and body weight measurement were conducted weekly. (b) Diets and experimental design -44ALPE12/C During the adaptation period, rats were fed the AIN-93 synthetic diet for one week. Subsequently, all rats were fed the AIN-93 diet supplemented with 3% cholesterol for another week (High cholesterol diet; HCD). During the experimental period the rats were divided into 4 groups and were fed different diets for 4 weeks according to Table 3. The diets were prepared in pellet form by Special Diet Services, Essex, England.

Table 3.The High Cholesterol Diet (HCD) fed to the rats was supplemented for each group as described below.

HCD +1%OC (MDOC) +0.5% Sterol Combination - 1% OC (MDOC) - 1% OC (MDOC) - - 0.5% Sterol 0.5% Sterol (c) Test compounds.

Oxidised cellulose (OC) MDOC was prepared as described above. Generic sterols (Gen) were purchased from commercially available products. (d) Statistics One-way ANOVA are used to determine treatment differences (SigmaStat version 1.0). Differences among means will be inspected using appropriate multiple comparisons test and were considered significant at P<0.05. (e) Results Animal weights No difference across treatments in final body weights was observed (p > 0.05). This indicates that the treatment ingredients did not alter the food consumption of the rats in the treatment groups compared to the control group (HCD). (Figure 6) IfcofiO750 _ 45 _ AI.PF.I2/C Table 4. Weight gain during the course of the study for the control group and for the three treatment groups.

Day HCD +1% OC (MDOC) +0.5% STEROL +1%OC (MDOC) & 0.5% STEROL -14 161+7.2 163+7.7 162+8.3 165+6.3 0 305+16.1 304+15.8 305+18.1 305+15.6 7 367+21.1 361+17.0 367+23.6 360+18.9 14 420+25.6 413+27.7 417+31.9 405+23.7 21 467+30.3 458+34.6 462+36.8 447+27.6 28 480+33.5 477+36.6 481+39.2 460+29.4 Example 5 - combination therapy changes in LDL cholesterol (a) Analytical measurements.

At the end of the experiment, animals (from Example 4) were fasted for 12 h and blood collected by cardiac puncture under terminal anaesthesia (anesthetized with ketamine (75mg/kg) and xylazine (lOmg/kg) administered intraperitoneally). Blood was allowed to clot and then transferred to a refrigerated centrifuge and centrifuged at 2,500 rpm for 10 minutes at 2-8°C. The collected serum was frozen at -20 °C until analysis. (b) Determination of serum lipid factors Serum LDL and HDL-cholesterol were measured enzymatically by using commercial available kits (EZ LDL™, EZ HDL™: Cholesterol kits, Trinity Biotech). Total cholesterol was measured using the commercially available kits from Human, Germany. Serum glucose was determined by colorimetric glucose oxidase assay (Sigma-Aldrich).

Triglycerides were measured with a commercially available kit from Sigma-Aldrich. (c) Statistics -46lfc0607A0 ALPE12/C One-way ANOVA are used to determine treatment differences (SigmaStat version 1.0). Differences among means will be inspected using appropriate multiple comparisons test and were considered significant at P<0.05. (d) Statistical Analysis Table 5 The effect of the dietary supplementation of a High Cholesterol Diet (HCD) with 1% oxidized cellulose (OC) MDOC and 0.5% sterol (w/w) on the serum LDL cholesterol concentrations in rats (n=20/gp) fed these diets.

Group HCD +1%OC (MDOC) +0.5% Sterol +1%OC (MDOC) +0.5% Sterol Mean ± SD 69.65 ± 6.50 61.63 + 7.38 60.77+7.12 54.62+6.71 % HCD 100 88 87 78 ANOVA The differences in the mean values among the treatment groups are greater than would be expected by chance; there is a statistically significant difference (p<0.001).

Equal Variance Test: Passed 0.665 All Pairwise Multiple Comparison Procedures (Student-Newman-Keuls Method): Comparison p HCD vs. +1 %OC (MDOC) & 0.5% Sterol <0.001 HCD vs.+0.5% Sterol <0.001 HCD vs. +1 %OC (MDOC) 0.001 +1 %OC (MDOC) vs. +1 %OC (MDOC) & 0.5% Sterol 0.011 +0.5% Sterol vs. + 1%OC (MDOC) & 0.5%Sterol 0.009 -47ALPE12/C (e) Results Table 5 illustrates an example of a combination therapy of the present invention. The example comprises four groups of subjects and the effectes on LAL (bad) cholesterol reduction. The four groups are: a control group that received no agent; a group that receive 1% (w/w) oxidised cellulose (OC) MDOC; a group that received 0.5% (w/w) sterol; and a final group that received both the 1% (w/w) OC MDOC and the 0.5% (w/w) sterol.

As can be seen from Figure 7, the combination of 0.1% oxidised cellulose and 0.5% sterol (w/w) caused a greater decrease in the level of LDL (bad) cholesterol than either 0.1 % oxidised cellulose or 0.5% sterol on their own. Therefore the combination of OC (MDOC) and sterol is more effective as an anti-atherosclerosis or an anti-hyperlipidemic.

Example 6 - combination therapy, changes in total cholesterol (a) Analytical measurements.

At the end of the experiment, animals (from example 4) were fasted for 12 h and blood collected by cardiac puncture under terminal anaesthesia (anesthetized with ketamine (75mg/kg) and xylazine (lOmg/kg) administered intraperitoneally). Blood was allowed to clot and then transferred to a refrigerated centrifuge and centrifuged at 2,500 rpm for 10 minutes at 2-8°C. The collected serum was frozen at -20 °C until analysis. (b) Determination of serum lipid factors Serum LDL and HDL-cholesterol were measured enzymatically by using commercial available kits (EZ LDL™, EZ HDL™: Cholesterol kits, Trinity Biotech). Total cholesterol was measured using the commercially available kits from Human, Germany. Serum glucose will be determined by colorimetric glucose oxidase assay (SigmaAldrich). Triglycerides were measured with a commercially available kit from SigmaAldrich. -48ALPE12/C ItO 6° Τ'® (c) Statistics One-way ANOVA are used to determine treatment differences (SigmaStat version 1.0). Differences among means will be inspected using appropriate multiple comparisons test and were considered significant at P<0.05. (d) Statistical Analysis Table 6 The effect of the dietary supplementation of a High Cholesterol Diet (HCD) with 1% oxidized cellulose (OC) MDOC and 0.5% sterol (w/w) on the serum Total cholesterol concentrations in rats (n=20/gp) fed these diets.

Group HCD +1%OC (MDOC) +0.5% Sterol +1%OC (MDOC) +0.5% Sterol Mean ± SD 81.43 ± 13.17 74.05 ± 13.34 74.09+ 13.73 69.70+ 10.62 % HCD 100 91 91 86 ANOVA The differences in the mean values among the treatment groups are greater than would be expected by chance; there is a statistically significant difference (p=0.041).

Equal Variance Test: Passed (0.395) All Pairwise Multiple Comparison Procedures (Student-Newman-Keuls Method): Comparison P HCD vs. +1 %OC (MDOC) & 0.5% sterol 0.025 All other comparisons are not significant. (e) Results -49A1.REI2/C Table 6 illustrates an example of a combination therapy of the present invention. The example comprises four groups of subjects and the effectes on LAL (bad) cholesterol reduction. The four groups are: a control group that received no agent; a group that receive 1% (w/w) oxidised cellulose (OC) MDOC; a group that received 0.5% (w/w) sterol; and a final group that received both the 1% (w/w) OC (MDOC) and the 0.5% (w/w) sterol.

As can be seen from Figure 8, the combination of 0.1% oxidised cellulose and 0.5% sterol (w/w) caused a greater decrease in the level of total cholesterol than either 0.1 % oxidised cellulose or 0.5% sterol on their own. Therefore the combination of OC (MDOC) and sterol is more effective as an anti-atherosclerosis or an anti-hyperlipidemic.

Example 7 - Glycaemic management (a) Animals.

Rats with a body weight of approximately 175g were obtained from Charles River Laboratories, Ballina, Ireland. After 1 wk of acclimation, 30 rats were allocated to 3 similar groups (n = 10 per group) on the basis of their body weight. The rats were housed in groups of two and three in standard cages with autoclaved dust-free processed sawdust as bedding material. The rats were housed in an environmentally controlled room (temperature 22 ± 3°C) with a 12-h light:dark cycle. Throughout the study, the rats had free access to food and drinking water. Clinical observation and body weight measurement were conducted once a week. (b) Diets and experimental design During the adaptation period, rats were fed the AIN-93 synthetic diet for one week. Subsequently, all rats were fed the AIN-93 diet supplemented with 1% cholesterol for another week (High cholesterol diet; HCD). During the experimental period the rats were divided into three groups and were fed different diets for 25 days. The first group was fed the AIN-93 diet supplemented with 1% cholesterol. The second and third groups IfcofcO 71 ** -50ALPE12/C were also fed the ΑΓΝ-93 diet supplemented with 1% cholesterol and 1% or 5% oxidized cellulose respectively. The diets were prepared in pellet form by Special Diet Services, Essex, England. (c) Test compounds.

OC (MDOC) was prepared as described above. (dj Statistics One-way ANOVA was used to determine treatment differences (SigmaStat version 1.0). 10 Differences among means were inspected using Tukey multiple comparisons test and were considered significant at P<0.05. Jf normality test failed non-parametric tests were used. The Kruskal-Wallis non-parametric ANOVA test was performed with Dunn’s post ANOVA multiple comparisons test. (e) Results Animal weights The three treatment groups started with similar mean body weights that were not statistically different from each other. These mean body weights ranged from 256.9+49.5 to 273.4+16.5. A summary of the body weights is provided in Figure 9 and the weight gain over the course of the study is also presented.

Table 7 Weight gain during the course of the study for the control group and for the two treatment groups.

Day Control 1% MDOC 5% MDOC 0 273.4+16.5 272.5+13.2 256.9+49.5 7 341.4+25.9 344.5+20.2 339.3+17.6 14 384.3+34.1 387.9+23.4 388.07+21.0 21 405.5+42.4 423.4+26.4 434.3+23.4 -51 ALPEJ2/C 4l9.4±39.7 428.5±28.9 436.1±27.0 All rats gained weight during the study period with no differences across treatments in final body weights (p > 0.05). This indicates that the treatment ingredient did not alter the food consumption of the rats in the treatment groups compared to the control group.

Example 8 - Glycaemic management fa) Analytical measurements.

At the end of the experiment, animals were fasted for 12 h and blood was collected by 10 cardiac puncture under terminal anaesthesia (anesthetized with ketamine (75mg/kg) and xylazine (lOmg/kg) administered intraperitoneally). Blood was allowed to clot and was then transferred to a refrigerated centrifuge and centrifuged at 2,500 rpm for 10 minutes at 2-8°C. The collected serum was then frozen at -20 °C until analysis. (b) Statistics.

One-way ANOVA was used to determine treatment differences (SigmaStat version I.0). Differences among means were inspected using Tukey multiple comparisons test and were considered significant at P<0.05. If normality test failed non-parametric tests were used. The Kruskal-Wallis non-parametric ANOVA test was performed with Dunn’s post ANOVA multiple comparisons test. (c) Results Changes in fasting serum glucose The results of the fasting serum glucose levels are presented in Figure 10. These results 25 indicated a reduction in the serum glucose in the rats fed a HCD supplemented with MDOC. The group of rats fed the HCD supplemented with 5% MDOC (w/w) significantly reduced the glucose concentration in the serum to 90.74% of the control value (p<0.00I). -52AI.PEI2/C Ifcoeo 7 7 ο Fasting blood glucose concentrations in the rats fed the HCD supplemented with 5% MDOC was significantly decreased (~9%) compared to the control group. This result may have very interesting possibilities for management of diabetes meliitus. CHD is a long-term complication of diabetes meliitus. The American Diabetes Association recognized the relationship of diabetes and CHD. It has documented goals of which optimal serum lipid levels, as well as maintenance of near-normal blood glucose levels are of primary importance (ADA, 2000). The potential of MDOC to manage both the glucose and serum lipid levels may be of great significance to diabetes meliitus sufferers.

The examples herein can be performed by substituting the generically or specifically described therapeutic compounds or inert ingredients for those used in the preceding examples.

The invention is not limited to the embodiments hereinbefore described which may be varied in detail. Such variations are not to be regarded as a departure from the spirit and scope of the present invention, and all such modifications and equivalents as would be obvious to one skilled in the art are intended to be included within the scope of the claims. -53ALPEI2/C References Davies, MJ., Woolf, N., Rowles, P.M., Pepper, J., 1988. Morphology of the endothelium over atherosclerotic plaques in human coronary arteries. British Heart Journal 60, 459 -464.

DeGraba, T.J., 1997. Expression of inflammatory mediators and adhesion molecules in human atherosclerotic plaque. Neurology 49, S15 - S19.

Farmer, J.A., 2000. Pleiotropic effects of statins. Current Atherosclerosis Reports 2, 208 -217.

Gundersen, H.J., Bagger, P., Bendtsen, T.F., Evans, S.M., Korbo, L., Marcussen, N., Moller, A., Nielsen, K., Nyengaard, J.R., Pakkenberg, B., et al., 1988. The new stereological tools: disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis. Apmis 96, 857 - 881. liyama, K., Hajra, L., liyama, M., Li, H., DiChiara, M., Medoff, B.D., Cybulsky, M.I., 1999. Patterns of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 expression in rabbit and mouse atherosclerotic lesions and at sites predisposed to lesion formation. Circulation Research 85, 199-207.

LaRosa, J.C., 2001. Pleiotropic effects of statins and their clinical significance. American Journal of Cardiology 88, 291 - 293.

Laufs, U., La Fata,V., Plutzky, J., Liao, J.K., 1998. Upregulation of endothelial nitric oxide synthase by HMG CoA reductase inhibitors.

Nachtigal, P., Semecky, V., Gojova, A., Kopecky, M., Benes, V., Juzkova, R., 2002. The application of stereological methods for the quantitative analysis of the atherosclerotic lesions in rabbits. Image Analysis and Stereology 21, 165 - 174.

Nachtigal, P., Semecky, V., Kopecky, M., Gojova, A., Solichova, D., Zdansky, P., Zadak, Z., 2004. Application of stereological methods for the quantification of VCAM-1 and 1CAM-I expression in early stages of rabbit atherogenesis. Pathology Research and Practice 200, 219 - 229.

Nakashima, Y., Plump, A.S., Raines, E.W., Breslow, J.L., Ross, R., 1994. ApoE-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree. Arteriosclerosis and Thrombosis 14, 133 - 140. -54AI.PF.I2/C Nakashima, Y., Raines, E.W., Plump, A.S., Breslow, J.L., Ross, R., 1998. Upregulation of VCAM-1 and ICAM-1 at atherosclerosis-prone sites on the endothelium in the ApoEdeficient mouse. Arteriosclerosis Thrombosis and Vascular Biology 18, 842 851.

Reddick, R.L., Zhang, S.H., Maeda, N., 1994. Atherosclerosis in mice lacking apo E. Evaluation of lesional development and progression. Arteriosclerosis and Thrombosis 14, 141 - 147.

Ross, R., 1999. Atherosclerosis—an inflammatory disease. New England Journal of Medicine 340, 115-126.

Sparrow, C.P., Burton, C.A., Hernandez, M., Mundt, S., Hassing, H., Patel, S., Rosa, R., Hermanowski-Vosatka, A., Wang, P.R., Zhang, D., Peterson, L., Detmers, P.A., Chao, Y.S., Wright, S.D., 2001. Simvastatin has anti-inflammatory and antiatherosclerotic activities independent of plasma cholesterol lowering. Arteriosclerosis Thrombosis and Vascular Biology 21, 115- 121.

Springer,T.A., 1994.Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell 76, 301 - 314.

Vaughan, CJ., Murphy, M.B., Buckley, B.M., 1996. Statins do more than just lower cholesterol. Lancet 348, 1079- 1082.

Weibel, E., 1979. Stereological methods. Practical Methods for Biology Morphometry, vol. 1. Academic Press.

Zhang, S.H., Reddick, R.L., Piedrahita, J.A., Maeda, N., 1992. Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E. Science 258,

Claims

1. A composition comprising a first component selected from a biocompatible anionic polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof, and an intermolecular complex thereof, and a second component comprising an anti-lipemic agent.

2. A composition as claimed in claim 1 wherein the second component is selected from a sterol, an esterified and/or hydrogenated sterol, a stand and a statin.

3. A composition as claimed in claim 1 wherein the second component comprises a sterol.

4. A composition as claimed in claim 2 wherein the statin is a 3-Hydroxy-3methylglutaryl CoA (HMG CoA) reductase inhibitor.

5. A composition as claimed in claim I wherein the second component is selected from one or more of the group consisting of: a cholesteryl ester transfer protein (CETP) inhibitor; a enterocyte cholesterol transporter inhibitor; nicotinic acid; niacin; a peroxisome proliferator-activated activator (PPAR) agonist; fish oil and soluble fibre such as psyllium, glucomannam, HPMC cellulose or glucan.

6. A composition as claimed in any one of the preceding claims for oral administration.

7. A composition as claimed in claim 6 wherein the composition is in the form of a tablet, pellet, capsule, granule or microsphere.

8. A composition as claimed in claim 6 wherein the composition is in a form suitable for incorporation into foods, beverages, nutraceutical or pharmaceuticals. AI.PEI2/C

9. A composition as claimed in any preceding claim wherein the first component is derived from a starch, cellulose or gum, or is of microbial origin.

10. A composition as claimed in any preceding claim wherein the first component comprises a microdispersed cellulose or derivative thereof.

11. A composition as claimed in any one of the preceding claims wherein the first component is prepared by partial or complete hydrolysis and neutralisation of a PAGA containing material.

12. A composition as claimed in any one of the preceding claims wherein the first component interacts with biomolecules in the fluid media of the gastrointestinal tract.

13. A composition as claimed in claim 1 wherein the polyanhydroglucuronic acid and salt thereof contain in their polymeric chain from 8 to 30 percent by weight of carboxyl groups, at least 80 percent by weight of these groups being uronic groups, at most 5 percent by weight of carbonyl groups, and at most 0.5 percent by weight of bound nitrogen.

14. A composition as claimed in claim 13 wherein the polyanhydroglucuronic acid and salt thereof contain in their polymeric chain at most 0.2 percent by weight of bound nitrogen.

15. A composition as claimed in claim 13 wherein the molecular mass of the polymeric chain of the polyanhydroglucuronic acid and salt thereof is from I X 10 3 to 3 X 10 5 Daltons. ALPKI2/C

16. A composition as claimed in claim 13 wherein the molecular mass of the polymeric chain of the anionic component ranges from 5 X 10 3 to 1.5 X 10 s Daltons.

17. A composition as claimed in claim 13 wherein the content of carboxyl groups is in the range of from 12 to 26 percent by weight, at least 95 percent of these groups being uronic groups.

18. A composition as claimed in claim 13 wherein the polyanhydroglucuronic acid and salt thereof contain at most 1 percent by weight of carbonyl groups.

19. A composition as claimed in claim 18 wherein each carbonyl group is an intra- or intermolecular 2,6 or 3,6 hemiacetal, a 2,4-hemiaIdal or a C2-C3 aldehyde.

20. A composition as claimed in claim I wherein the biocompatible intermolecular polymer complex is a complex of: an anionic component comprising polyanhydroglucuronic acid or salt, which is that of a partially or completely hydrolysed and oxidativeenvironment hydrolysed polyanhydroglucuronic acid containing material; and a non protein cationic component comprising a linear or branched natural, semi-synthetic or synthetic oligomer or polymer.

21. A composition as claimed in claim 20 wherein at least 5% of the basic structural units of the anionic component are glucuronic acid. -58A1.PE12/C

22. A composition as claimed in claim 20 wherein the cationic component contains nitrogen that either carries a positive charge or wherein a positive charge is induced by contact with the polysaccharidic anionic component.

23. A composition as claimed in claim 22 wherein the cationic component is a member selected from the group consisting of a derivative of acrylamide, a derivative of methacrylamide, a copolymer of acrylamide and a copolymer of methacrylamide.

24. A composition as claimed in claim 22 wherein the cationic component is a member selected from the group consisting of polyacrylamide, a copolymer of hydroxyethylmethacrylate and hydroxypropylmethacrylamide, and a copolymer of acrylamide, butylacrylate, maleicanhydride and methylmethacrylate.

25. A composition as claimed in claim 20 wherein the cationic component is a cationised natural polysaccharide.

26. A composition as claimed in claim 25 wherein the polysaccharide is a starch, cellulose or gum.

27. A composition as claimed in claim 26 wherein the gum is guargumhydroxypropyltriammonium chloride.

28. A composition as claimed in claim 10 wherein the cationic component is a synthetic or semi-synthetic polyamino acid.

29. A composition as claimed in claim 28 wherein the cationic component is a member selected from the group consisting of polylysin, polyarginin and .alpha, beta.-poly-[N-(2-hydroxyethyl)-DL-aspartamide]. -59ALPEI2/C

30. A composition as claimed in claim 29 wherein the cationic component is a synthetic anti-fibrinolytic.

31. A composition as claimed in claim 30 wherein the anti-fibrinolytic is a hexad i methrindibromide.

32. A composition as claimed in claim 20 wherein the cationic component is a natural or semi-synthetic peptide.

33. A composition as claimed in claim 32 wherein the peptide is a member selected from the group consisting of a protamine, gelatine, fibrinopeptide, and a derivative of one of the foregoing.

34. A composition as claimed in claim 20 wherein the cationic component is an aminoglucane or derivative thereof.

35. A composition as claimed in claim 34 wherein the aminoglucane is fractionated chitin or its de-acetylated derivative chitosan.

36. A composition as claimed in claim 34 wherein the aminoglucane is of microbial origin or is isolated from an arthropod shell.

37. A composition as claimed in claim 34 wherein the cationic component is gelatin.

38. A composition as claimed in claim 34 wherein the cationic component is chitosan.

39. Use of a biocompatible anionic polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof or an intermolecular complex thereof, in the preparation of a medicament for the treatment of inflammation. -60AI.PEI2/C

40. Use as claimed in claim 39 wherein the inflammation is associated with atherosclerosis

41. Use as claimed in claim 39 wherein the medicament is for oral administration.

42. Use as claimed in claim 41 wherein the medicament is in the form of a tablet, pellet, capsule, granule or microsphere.

43. Use as claimed in claim 41 wherein the medicament is in a form suitable for incorporation into foods, beverages, nutraceutical and pharmaceuticals.

44. Use as claimed in any one of claims 39 to 43 wherein the polyanhydroglucuronic acid, a salt thereof, a copolymer thereof and an intermolecular complex thereof are derived from a starch, cellulose or gum, or are of microbial origin.

45. Use as claimed in any one of claims 39 to 44 wherein the PAGA comprises a microdispersed cellulose or derivative thereof.

46. Use as claimed in any one of claims 39 to 45 wherein said PAGA is prepared by partial or complete hydrolysis and neutralisation of a PAGA containing material.

47. Use as claimed in any one of claims 39 to 46 wherein the PAGA interacts with biomolecules in the fluid media of the gastrointestinal tract.

48. Use as claimed in claim 39 wherein the polyanhydroglucuronic acid and salt thereof contain in their polymeric chain from 8 to 30 percent by weight of carboxyl groups, at least 80 percent by weight of these groups being uronic groups, at most 5 percent by weight of carbonyl groups, and at most 0.5 percent by weight of bound nitrogen. -61 ALPEI2/C

49. Use as claimed in claim 48 wherein the polyanhydroglucuronic acid and salt thereof contain in their polymeric chain at most 0.2 percent by weight of bound nitrogen.

50. Use as claimed in claim 48 wherein the molecular mass of the polymeric chain of the polyanhydroglucuronic acid and salt thereof is from 1 X 10 3 to 3 X IO 3 Daltons.

51. Use as claimed in claim 48 wherein the molecular mass of the polymeric chain of the anionic component ranges from 5 X 10 3 to 1.5 X 10 5 Daltons.

52. Use as claimed in claim 48 wherein the content of carboxyl groups is in the range of from 12 to 26 percent by weight, at least 95 percent of these groups being uronic groups.

53. Use as claimed in claim 48 wherein the polyanhydroglucuronic acid and salt thereof contain at most 1 percent by weight of carbonyl groups.

54. Use as claimed in claim 53 wherein each carbonyl group is an intra- or intermolecular 2,6 or 3,6 hemiacetal, a 2,4-hemialdal or a C2-C3 aldehyde.

55. Use as claimed in claim 39 wherein the biocompatible intermolecular polymer complex is a complex of: an anionic component comprising polyanhydroglucuronic acid or salt, which is that of a partially or completely hydrolysed and oxidativeenvironment hydrolysed polyanhydroglucuronic acid containing material; and ALPEI2/C a non protein cationic component comprising a linear or branched natural, semi-synthetic or synthetic oligomer or polymer.

56. Use as claimed in claim 55 wherein at least 5% of the basic structural units of the anionic component are glucuronic acid.

57. Use as claimed in claim 55 wherein the cationic component contains nitrogen that either carries a positive charge or wherein a positive charge is induced by contact with the polysaccharidic anionic component.

58. Use as claimed in claim 58 wherein the cationic component is a member selected from the group consisting of a derivative of acrylamide, a derivative of methacrylamide, a copolymer of acrylamide and a copolymer of methacrylamide.

59. Use as claimed in claim 55 wherein the cationic component is a member selected from the group consisting of polyacrylamide, a copolymer of hydroxyethylmethacrylate and hydroxypropylmethacrylamide, and a copolymer of acrylamide, butylacrylate, maleicanhydride and methylmethacrylate.

60. Use as claimed in claim 55 wherein the cationic component is a cationised natural polysaccharide.

61. Use as claimed in claim 60 wherein the polysaccharide is a starch, cellulose or gum.

62. Use as claimed in claim 61 wherein the gum is guargumhydroxypropyltriammonium chloride.

63. Use as claimed in claim 47 wherein the cationic component is a synthetic or semisynthetic polyamino acid. AI.PEI2/C

64. Use as claimed in claim 63 wherein the cationic component is a member selected from the group consisting of polylysin, polyarginin and .alpha, beta.-poly-[N-(2hydroxyethyl)-DL-aspartamide],

65. Use as claimed in claim 64 wherein the cationic component is a synthetic antifibrinolytic.

66. Use as claimed in claim 65 wherein the anti-fibrinolytic is a hex ad i methri nd i brom ide.

67. Use as claimed in claim 55 wherein the cationic component is a natural or semisynthetic peptide.

68. Use as claimed in claim 67 wherein the peptide is a member selected from the group consisting of a protamine, gelatine, fibrinopeptide, and a derivative of one of the foregoing.

69. Use as claimed in claim 55 wherein the cationic component is an aminoglucane or derivative thereof.

70. Use as claimed in claim 69 wherein the aminoglucane is fractionated chitin or its de-acetylated derivative chitosan.

71. Use as claimed in claim 69 wherein the aminoglucane is of microbial origin or is isolated from an arthropod shell.

72. Use as claimed in claim 55 wherein the cationic component is gelatin.

73. Use as claimed in claim 55 wherein the cationic component is chitosan. ALPEI2/C

74. Use of a biocompatible anionic polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof or an intermolecular complex thereof, in the preparation of a medicament for maintaining a patient’s blood glucose concentration in a physiological range.

75. Use as claimed in claim 74 wherein the patient has type 2 diabetes mellitus.

76. Use as claimed in claim 74 wherein the composition/medicament is for oral administration. ΊΊ. Use as claimed in claim 76 wherein the composition/medicament is in the form of a tablet, pellet, capsule, granule or microsphere.

77. 78. Use as claimed in claim 76 wherein the composition/medicament is in a form suitable for incorporation into foods, beverages, nutraceutical or pharmaceuticals.

78. 79. Use as claimed in any one of claims 76 to 78 wherein the polyanhydroglucuronic acid, a salt thereof, a copolymer thereof, and an intermolecular complex thereof are derived from a starch, cellulose or gum, or are of microbial origin.

79. 80. Use as claimed in any one of claims 76 to 79 wherein the PAGA comprises a microdispersed cellulose or derivative thereof.

80. 81. Use as claimed in any one of claims 76 to 80 wherein said PAGA prepared by partial or complete hydrolysis and neutralisation of a PAGA containing material.

81. 82. Use as claimed in any one of claims 76 to 81 wherein the PAGA interacts with biomolecules in the fluid media of the gastrointestinal tract. ALPE12/C

82. 83. Use as claimed in claim 76 wherein the polyanhydroglucuronic acid and salt thereof contain in their polymeric chain from 8 to 30 percent by weight of carboxyl groups, at least 80 percent by weight of these groups being uronic groups, at most 5 percent by weight of carbonyl groups, and at most 0.5 percent by weight of bound nitrogen.

83. 84. Use as claimed in claim 83 wherein the polyanhydroglucuronic acid and salt thereof contain in their polymeric chain at most 0.2 percent by weight of bound nitrogen.

84. 85. Use as claimed in claim 83 wherein the molecular mass of the polymeric chain of the polyanhydroglucuronic acid and salt thereof is from 1 X 10 3 to 3 X IO 5 Daltons.

85. 86. Use as claimed in claim 83 wherein the molecular mass of the polymeric chain of the anionic component ranges from 5 X 10 3 to 1.5 X 10 5 Daltons.

86. 87. Use as claimed in claim 83 wherein the content of carboxyl groups is in the range of from 12 to 26 percent by weight, at least 95 percent of these groups being uronic groups.

87. 88. Use as claimed in claim 83 wherein the polyanhydroglucuronic acid and salt thereof contain at most 1 percent by weight of carbonyl groups.

88. 89. Use as claimed in claim 88 wherein each carbonyl group is an intra- or intermolecular 2,6 or 3,6 hemiacetal, a 2,4-hemialdal or a C2-C3 aldehyde.

89. 90. Use as claimed in claim 76 wherein the biocompatible intermolecular polymer complex is a complex of: an anionic component comprising polyanhydroglucuronic acid or salt, which is that of a partially or completely hydrolysed and oxidativeenvironment hydrolysed polyanhydroglucuronic acid containing material; and ALPEI2/C a non protein cationic component comprising a linear or branched natural, semi-synthetic or synthetic oligomer or polymer.

90. 91. Use as claimed in claim 90 wherein at least 5% of the basic structural units of the anionic component are glucuronic acid.

91. 92. Use as claimed in claim 90 wherein the cationic component contains nitrogen that either carries a positive charge or wherein a positive charge is induced by contact with the polysaccharidic anionic component.

92. 93. Use as claimed in claim 92 wherein the cationic component is a member selected from the group consisting of a derivative of acrylamide, a derivative of methacrylamide, a copolymer of acrylamide and a copolymer of methacrylamide.

93. 94. Use as claimed in claim 92 wherein the cationic component is a member selected from the group consisting of polyacrylamide, a copolymer of hydroxyethylmethacrylate and hydroxypropylmethacrylamide, and a copolymer of acrylamide, butyl acrylate, maleicanhydride and methylmethacrylate.

94. 95. Use as claimed in claim 90 wherein the cationic component is a cationised natural polysaccharide.

95. 96. Use as claimed in claim 95 wherein the polysaccharide is a starch, cellulose or gum. ALPEI2/C

96. 97. Use as claimed in claim 96 wherein the gum is guargumhydroxypropyltriammonium chloride.

97. 98. Use as claimed in claim 80 wherein the cationic component is a synthetic or semisynthetic polyamino acid.

98. 99. Use as claimed in claim 98 wherein the cationic component is a member selected from the group consisting of polylysin, polyarginin and .alpha, beta.-poly-[N-(2hydroxyethyl)-DL-aspartamide],

99. 100. Use as claimed in claim 99 wherein the cationic component is a synthetic antifibrinolytic.

100. 101 .Use as claimed in claim 100 wherein the anti-fibrinolytic is a hexadimethrindibromide.

101. 102. Use as claimed in claim 90 wherein the cationic component is a natural or semisynthetic peptide.

102. 103. Use as claimed in claim 102 wherein the peptide is a member selected from the group consisting of a protamine, gelatine, fibrinopeptide, and a derivative of one of the foregoing.

103. 104. Use as claimed in claim 90 wherein the cationic component is an aminoglucane or derivative thereof.

104. 105. Use as claimed in claim 104 wherein the aminoglucane is fractionated chitin or its de-acetylated derivative chitosan. -68ΑΙΤΕΙ 2/C

105. 106. Use as claimed in claim 104 wherein the aminoglucane is of microbial origin or is isolated from an arthropod shell.

106. 107. Use as claimed in claim 90 wherein the cationic component is gelatin. 5

107. 108.Use as claimed in claim 90 wherein the cationic component is chitosan.