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WO2007036920A2 - Utilisation d'acide polyanhydroglucronique - Google Patents

Utilisation d'acide polyanhydroglucronique Download PDF

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WO2007036920A2
WO2007036920A2 PCT/IE2006/000104 IE2006000104W WO2007036920A2 WO 2007036920 A2 WO2007036920 A2 WO 2007036920A2 IE 2006000104 W IE2006000104 W IE 2006000104W WO 2007036920 A2 WO2007036920 A2 WO 2007036920A2
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composition
component
cationic component
percent
salt
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PCT/IE2006/000104
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WO2007036920A3 (fr
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Jiri Briestensky
Ivan Santar
Vladimir Semecky
Petr Nachtigal
Anthony Richardson
Keith Real
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Alltracel Development Services Limited
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Publication of WO2007036920A3 publication Critical patent/WO2007036920A3/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/74—Synthetic polymeric materials
    • A61K31/785—Polymers containing nitrogen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/716—Glucans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/716—Glucans
    • A61K31/717—Celluloses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/716—Glucans
    • A61K31/718—Starch or degraded starch, e.g. amylose, amylopectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/716—Glucans
    • A61K31/722—Chitin, chitosan
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/736—Glucomannans or galactomannans, e.g. locust bean gum, guar gum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/74—Synthetic polymeric materials
    • A61K31/765—Polymers containing oxygen
    • A61K31/78—Polymers containing oxygen of acrylic acid or derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system

Definitions

  • the invention relates to use of polyanhydroglucuronic acid (PAGA), a salt thereof, a copolymer thereof or an intermolecular complex thereof and a composition containing PAGA, a salt thereof, a copolymer thereof and an intermolecular complex thereof.
  • PAGA polyanhydroglucuronic acid
  • Cardiovascular disease Cardiovascular disease still remains the leading cause of death and disability in the western world.
  • NCEP National Cholesterol Education Program
  • LDL low density lipoprotein
  • WHO World Health Organization
  • 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.
  • LDL-C low density lipoprotein cholesterol
  • HDL-C high density lipoprotein 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).
  • Cholesterol 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 are particles that carry lipids and proteins in the blood and gut. There are five classes of lipoproteins:
  • VLDL Very low-density lipoprotein
  • IDL Intermediate-density lipoprotein
  • LDL Low-density lipoprotein
  • HDL High-density lipoprotein
  • LDL is 'bad' cholesterol and is the main cause of harmful fatty buildup in arteries.
  • 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 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).
  • VCAM-I vascular cell adhesion molecule
  • IAM-I intercellular cell adhesion molecule
  • ICAM-I is expressed strongly on the endothelium overlying atheromatous plaque in human coronary and carotid arteries (DeGraba, 1997), hypercholesterolaemic rabbits (Iiyama et al., 1999), and apoE "7" (Nakashima et al., 1998) and LDLR “7" mice (Iiyama et al., 1999), although it is expressed in virtually all endothelial cells.
  • statin class of drugs was the development of the statin class of drugs.
  • HMG-CoA 3 -hydroxy-3 -methyl glutaryl coenzyme A reductase
  • statins exert beneficial vascular effects that are independent of their cholesterol lowering potencies (Farmer, 2000; LaRosa, 2001).
  • 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.
  • statins target hepatocytes and inhibit HMG-CoA reductase (3-hydroxy-3-methyl-glutaryl coenzyme A reductase, E. C.1.1.1.34), the enzyme that converts HMG-CoA into mevalonic acid, a cholesterol precursor.
  • HMG-CoA reductase 3-hydroxy-3-methyl-glutaryl coenzyme A reductase, E. C.1.1.1.34
  • the change in conformation at the active site makes these drugs very effective and specific.
  • statins to HMGCoA reductase 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 density- VLDL 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 BfE.
  • Statins have a modest effect on HDL increase, and no influence on lipoprotein(s) concentration.
  • Plant sterols are poorly absorbed in the intestine (0.4-3.5%), while absorption of plant stands (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.
  • 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 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.
  • Bile acid sequestrants were the first class of antihyperlipidemic drugs developed to lower LDL cholesterol levels.
  • 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 7 ⁇ -hydroxylase, HMG-CoA reductase, and phosphatidic acid phosphatase.
  • bile acids returning to the liver through the enterohepatic pathway inhibit the activity of 7 ⁇ -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.
  • HMG-CoA reductase the rate-limiting enzyme in cholesterol synthesis.
  • total cholesterol levels do not 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.
  • 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.
  • 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.
  • VLD very-low- density
  • 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.
  • fibric acid derivatives i.e., fenofibrate and gemfibrozil
  • 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%.
  • 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 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.
  • 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.
  • high viscosity soluble fibres There are many examples of high viscosity soluble fibres.
  • the main commercially available soluble fibres are psyllium, glucomannan and ⁇ -glucan. These generally lead to an LDL reduction of about 6 to 7%.
  • 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 HDL- C, 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.
  • 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
  • Nicotinic acid 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. 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).
  • 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.
  • 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.
  • 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.
  • compositions 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.
  • 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.
  • the second component may be selected from a sterol, an esterified and/or hydrogenated sterol, a stanol and a statin.
  • the second component may comprise a sterol.
  • the second component maybe a satin such as a 3-Hydroxy-3- methylglutaryl 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.
  • CETP cholesteryl ester transfer protein
  • PPAR peroxisome proliferator-activated activator
  • fish oil and soluble fibre such as psyllium, glucomannam, HPMC cellulose or glucan.
  • the composition may be suitable for oral administration.
  • the composition may be in the form of a tablet, pellet, capsule, granule or microsphere.
  • 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.
  • 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.
  • 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.
  • PAGA biocompatible anionic polyanhydroglucuronic acid
  • 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.
  • PAGA biocompatible anionic polyanhydroglucuronic acid
  • the medicament maybe for oral administration.
  • the medicament maybe in the form of a tablet, pellet, capsule, granule or microsphere.
  • 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.
  • the medicament may further comprise an antilipemic agent.
  • the antilipemic agent may be a sterol, stanol, statin, esterified and/or hydrogenated sterol.
  • the statin may be a 3-Hydroxy-3-methylglutaryl CoA (HMG CoA) reductase inhibitor.
  • 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 proliferator- activated activator (PPAR) agonist; fish oil and soluble fibre such as psyllium, glucomannam, HPMC cellulose or glucan.
  • CETP cholesteryl ester transfer protein
  • PPAR peroxisome proliferator- activated activator
  • 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
  • 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.
  • 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 I X lO 3 to about 3 X 10 5 Daltons.
  • the molecular mass of the polymeric chain of the anionic component may range from about 5 X 10 3 to about 1.5 X 10 5 Daltons.
  • 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.
  • 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;
  • non protein cationic component comprising a linear or branched natural, semi-synthetic or synthetic oligomer or polymer.
  • 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 polysaccharide 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 acrylamide, butylacrylate, maleicanhydride and methylmethacrylate.
  • the cationic component may be a cationised natural polysaccharide.
  • the polysaccharide may be a starch, cellulose or gum.
  • the gum may be guargumhydroxypropyltriammonium chloride.
  • the cationic component may be a synthetic or semi-synthetic polyamino acid.
  • the cationic component may be a member selected from the group consisting of polylysin, polyarginin and .alpha, beta.-poly-[N-(2-hydroxyethyl)-DL-aspartamide].
  • 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.
  • 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.
  • 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.
  • PAGA biocompatible anionic polyanhydroglucuronic acid
  • 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, 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.
  • 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.
  • PAGA biocompatible anionic polyanhydroglucuronic acid
  • the invention also provides a method for improving the management of blood glucose and insulin for patients with type 2 diabetes mellitus 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.
  • 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.
  • PAGA biocompatible anionic polyanhydroglucuronic acid
  • 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.
  • 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.
  • PAGA biocompatible anionic polyanhydroglucuronic acid
  • 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.
  • 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.
  • PAGA biocompatible anionic polyanhydroglucuronic acid
  • the invention in particular involves the use of polyanhydroglucuronic acids, salts and intermolecular complexes (IMC) thereof.
  • polyanhydroglucuronic acid, salts and IMCs thereof as used herein also includes copolymers thereof, especially with anhydroglucose.
  • PAGA polyanhydroglucuronic acid, salts and IMCs thereof as used herein also includes copolymers thereof, especially with anhydroglucose.
  • PAGA Throughout the specification PAGA is also referred to as microdispersed oxidised cellulose (MDOC) available from Alltracel Pharma Limited. MDOC is a registered trademark.
  • polyanhydroglucuronic acids and salts thereof describe particular polyanhydroglucuronic acids and salts thereof and a method of preparing such compounds.
  • 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.
  • 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.
  • PAGA polyanhydroglucuronic acid
  • WO00/05269 describes particular intermolecular complexes (IMCs) of PAGA.
  • IMCs of PAGA in particular include those referred to in WO00/05269, the entire contents of which is incorporated herein by reference.
  • non-protein is intended to distinguish from what the applicants understand the commonly used definition in biochemistry of "protein” to be.
  • 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.
  • 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.
  • 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.
  • 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 1 below.
  • Fig. 1 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;
  • MDOC oxidized cellulose
  • Fig. 2 shows the immunohistochemical staining of endothelial expression of an inflammatory marker ICAM-I in (A) a control group, (B) a simvistatin group, (C) an atorvastatin group and (D) an oxidized cellulose (MDOC) group.
  • ICAM-I oxidized cellulose
  • Fig. 3 shows the immunohistochemical staining of endothelial expression of an inflammatory marker VCAM-I in A) a control group, (B) a simvistatin group, (C) an atorvastatin group and (D) an oxidized cellulose (MDOC) group.
  • a control group a control group
  • B simvistatin group
  • C an atorvastatin group
  • D an oxidized cellulose
  • MDOC oxidized cellulose
  • 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-I is significantly decreased in atorvastatin and MDOC treated mice in
  • 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
  • HCD high cholesterol diet
  • OC oxidised cellulose
  • sterol 0.5% sterol
  • HCD high cholesterol diet
  • OC oxidised cellulose
  • sterol 0.5% sterol
  • 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.
  • 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.
  • 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.
  • the oxidised polysaccharide material is in the form of biocompatible anionic polyanhydroglucuronic acid (PAGA), salt or intermolecular complex (IMC) thereof.
  • 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).
  • 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.
  • the polysaccharide material is polyanhydroglucuronic acid, biocompatible salts thereof, copolymers thereof or a biocompatible intermolecular complex polymer thereof.
  • the oxidised polysaccharide is derived from cellulose, starch, or gum, or is of microbial origin.
  • the PAGA 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.
  • PAGA Similar to hyaluronic acid (HA), the PAGA displays reducing ability in a biological environment, yet with a higher assimilable organic carbon (AOC) value.
  • 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.
  • GIT gastrointestinal tract
  • 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).
  • 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.
  • AIP atherogenic index of plasma
  • 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.
  • An advantage of the therapy of the 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, f ⁇ brates and niacin.
  • antilipemic compositions such as notably statins, sterols, stanols, CETP inhibitors, dietary fibre, f ⁇ brates and niacin.
  • 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.
  • composition of the present invention may be incorporated into foods, beverages and nutraceuticals, including, without limitation, the following:
  • Dairy Products such as cheeses, butter, milk and other dairy beverages, spreads and dairy mixes, ice cream and yoghurt;
  • Fat-Based Products such as margarines, spreads, mayonnaise, shortenings and dressings;
  • Cereal-Based Products comprising grains (for example, bread and pastas) however processed;
  • Confectionaries such as chocolate, candies, chewing gum, desserts, non-dairy toppings, sorbets, icings and other fillings
  • 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 BoostTM, and EnsureTM; and
  • Miscellaneous Products including eggs, processed foods such as soups, pre-prepared pasta sauces, pre-formed meals and the like.
  • 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.
  • the composition may be applied directly onto a food or into a beverage by the consumer prior to ingestion.
  • 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.
  • tablets plain and coated
  • capsules hard and soft, with or without additional coatings
  • powders powders
  • granules including effervescent granules
  • pellets including effervescent granules
  • microparticulates such as micellar, syrups, elixirs and drops
  • solutions such as micellar, syrups, elixirs
  • N-HANCE 3000 30 g were placed into a 5 1 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.
  • 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 1 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.
  • the isolated material thus prepared may further serve to prepare final products of the nonwoven type via a wet or dry process.
  • H2O2 anal.grade 30% (Lachema, a.s. Neratovice); gelatine (PhBs 1997)
  • 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 .
  • 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.
  • MDOC (CaTNa 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
  • 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 Fl/1 was 15 minutes at 20°C, and 8 minutes at 37°C.
  • 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.
  • H2O2 anal.grade 30% (Lachema, a.s. Neratovice) gelatine (PhBs 1997)
  • 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 (1O 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 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 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.
  • 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 Fl/1 was 17 minutes at 20°C, and 8 minutes at 37°C.
  • 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.
  • the product may be used as a) an embolisation agent, or b) an antilipemicum.
  • the product may be used as a) an embolisation agent, or b) an antilipemicum.
  • ApoE "A 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).
  • ApoE "7" 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).
  • mice Male apoE " ⁇ mice were weaned at 5 weeks of age and randomly subdivided into four groups.
  • chow diet standard laboratory diet
  • statins were added to the chow diet at the dosage of 10 mg/kg per day.
  • OCT Optimal Cutting Temperature
  • Serum lipoprotein fractions were prepared using NaCl density gradient ultracentrifugation (Beckman TL 100, Palo Alto, CA). The lipoprotein fractions were distinguished in the following density ranges: very low density lipoprotein (VLDL) ⁇ 1.006 g/ml; low density lipoprotein (LDL) ⁇ 1.063 g/ml; high density lipoprotein (HDL)>1.063 g/ml.
  • VLDL very low density lipoprotein
  • LDL low density lipoprotein
  • HDL high density lipoprotein
  • 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 cross- sections (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 0 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.
  • PBS phosphate buffered saline
  • 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.
  • 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
  • P is the number of test points hitting either the atherosclerotic lesion or positive immunostaining.
  • VCAM-I The area of Von Willebrand factor expression was considered as a total area of intact endothelium.
  • ICAM-I expression indicates the percentage of activated endothelial cells calculated as
  • estP areafx) *100% area (VonWill)
  • area (x) is the area of VCAM-I or ICAM-I in the endothelium and area (Von Will) is the area of Von Willebrand factor expression in the endothelium.
  • 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 Innovision, Incl, San Diego, Calif.) interfaced to a color camera (Toshiba 3 CCD) 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)).
  • a true color image analyzer Videometric 150; American Innovision, Incl, San Diego, Calif.
  • the abdominal aortas are rapidly excised, after injection of sodium pentobarbital, and placed in oxygenated Krebs-bicarbonate buffer. After removal of perivascular tissue, 3- mm 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 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.
  • apoE mice received normal chow diet or diet containing micro dispersed derivatives of oxidised cellulose (MDOC) 50 mg/kg/day or simvastain, atorvastatin 10mg/kg/day. Total cholesterol, VLDL, LDL, HDL and TAG were measured.
  • MDOC oxidised cellulose
  • Example 2 -Endothelial expression of VCAM-I in apoE ;" mouse model of atherosclerosis Members of the immunoglobulin superfamily of endothelial adhesion molecules, vascular cell adhesion molecule (VCAM-I) and intercellular cell adhesion molecule (ICAM-I) 5 strongly participate in leukocyte adhesion to the endothelium and play an important role in all stages of atherogenesis.
  • VCAM-I vascular cell adhesion molecule
  • IAM-I intercellular cell adhesion molecule
  • Hyperlipidemic apolipoprotein-E-deficient mice received normal chow diet or diet containing micro dispersed derivatives of oxidised cellulose (MDOC) 50 mg/kg/day or simvastain, atorvastatin 10mg/kg/day.
  • MDOC oxidised cellulose
  • atorvastatin 10mg/kg/day.
  • Total cholesterol, VLDL, LDL, HDL and TAG were measured and the endothelial expression of VCAM-I and ICAM-I was visualized and quantified by means of immunohistochemistry and stereology, respectively.
  • ICAM-I and VCAM-I in endothelium were 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-I staining was much stronger in all mice compared to the VCAM-I staining (Fig. 4).
  • the percentage of activated endothelial cells ICAM-I /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.
  • Example 3 VCAM expression in activated endothelial cells
  • 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).
  • MDOC is able to significantly decrease (77%) the expression of VCAM-I 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.
  • MDOC cardiovascular disease reduction
  • 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 O 2 " , 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.
  • the reduced accumulation of oxLDL might contribute to decreased numbers of macrophages in atheroma during lipid lowering and hence the atherosclerotic risk.
  • HCD High Cholesterol Diet
  • Oxidised cellulose (OC) MDOC was prepared as described above.
  • Generic sterols (Gen) were purchased from commercially available products.
  • Example 5 combination therapy changes in LDL cholesterol
  • mice from Example 4 were fasted for 12 h and blood collected by cardiac puncture under terminal anaesthesia (anesthetized with ketamine (75mg/kg) and xylazine (10mg/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 0 C until analysis.
  • Serum LDL and HDL-cholesterol were measured enzymatically by using commercial available kits (EZ LDLTM, EZ HDLTM: 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.
  • 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.
  • HCD High Cholesterol Diet
  • OC oxidized cellulose
  • sterol 0.5% sterol
  • 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.
  • Example 6 changes in total cholesterol
  • mice from example 4 were fasted for 12 h and blood collected by cardiac puncture under terminal anaesthesia (anesthetized with ketamine (75mg/kg) and xylazine (10mg/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 0 C until analysis.
  • Serum LDL and HDL-cholesterol were measured enzymatically by using commercial available kits (EZ LDLTM, EZ HDLTM: 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 (Sigma-
  • Triglycerides were measured with a commercially available kit from Sigma-
  • HCD High Cholesterol Diet
  • OC oxidized cellulose
  • sterol 0.5% sterol
  • Results 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.
  • MDOC and sterol is more effective as an anti-atherosclerosis or an anti-hyperlipidemic.
  • 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).
  • HCD High cholesterol diet
  • 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 were also fed the AIN-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.
  • 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.

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Abstract

L'invention concerne une composition comprenant un premier composant sélectionné parmi un acide polyanhydroglucuronique anionique biocompatible (PAGA), un sel de celui-ci, un copolymère de celui-ci et un complexe intermoléculaire de celui-ci et un second composant comprenant un agent anti-lipémique. L'invention concerne également l'utilisation d'un acide polyanhydroglucuronique anionique biocompatible (PAGA), d'un sel de celui-ci, d'un copolymère de celui-ci ou d'un complexe intermoléculaire de celui-ci dans la préparation d'un médicament destiné au traitement d'inflammation et l'utilisation d'un acide polyanhydroglucuronique anionique biocompatible (PAGA), d'un sel de celui-ci, d'un copolymère de celui-ci ou d'un complexe intermoléculaire de celui-ci dans la préparation d'un médicament destiné à maintenir la concentration en glucose dans le sang à un niveau physiologique.
PCT/IE2006/000104 2005-09-28 2006-09-28 Utilisation d'acide polyanhydroglucronique WO2007036920A2 (fr)

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US20020110531A1 (en) * 2001-02-14 2002-08-15 Steven Faust Cholesterol reducing composition and method of making the same
AU2003245750A1 (en) * 2002-02-14 2003-09-04 Alpenstock Holdings Limited Use of polyanhydroglucuronic acid comprising microdispersed oxidised cellulose as immunomodulator

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ITFI20120268A1 (it) * 2012-12-03 2014-06-04 Diopeite Consulting Ltd Composizioni farmaceutiche per il trattamento dell'obesita'
WO2014087317A1 (fr) * 2012-12-03 2014-06-12 Diopeite Consulting Ltd Compositions pharmaceutiques pour le traitement de l'obésité

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