WO1995029684A1 - Inhibition de l'atherosclerose au moyen d'antioxydants - Google Patents

Inhibition de l'atherosclerose au moyen d'antioxydants Download PDF

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Publication number
WO1995029684A1
WO1995029684A1 PCT/US1995/002658 US9502658W WO9529684A1 WO 1995029684 A1 WO1995029684 A1 WO 1995029684A1 US 9502658 W US9502658 W US 9502658W WO 9529684 A1 WO9529684 A1 WO 9529684A1
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vitamin
atherosclerosis
day
selenium
human
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PCT/US1995/002658
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English (en)
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Stephen Richard Behr
Dawn Colleen Schwenke
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Abbott Laboratories
Wake Forest University
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Priority to AU19388/95A priority Critical patent/AU1938895A/en
Publication of WO1995029684A1 publication Critical patent/WO1995029684A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/04—Sulfur, selenium or tellurium; Compounds thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/15—Vitamins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/16—Inorganic salts, minerals or trace elements
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present invention relates to the use of enterally ingested antioxidants for inhibiting atherosclerosis.
  • vitamin A Association between dietary changes and mortality rates: Israel 1949 to 1977: a trend-free regression model. Am J Clin Nutr 1981;34:1569-1583; Gey, et al . : Inverse Correlation Between Plasma Vitamin-E and Mortality from Ischemic Heart Disease in Cross-Cultural Epidemiology. Am J Clin Nutr 1991;53:S326-S334]: vitamin C [Katsouyanni, et al . : Diet and Peripheral Arterial Occlusive Disease - The Role of Polysaturated, Monosaturated. and Saturated Fatty Acids.
  • antioxidants or antioxidant nutrients caused a decrease in the plasma cholesterol concentration [Wojcicki , et al . : Effect of selenium and vitamin E on the development of experimental atherosclerosis in rabbits. Atherosclerosis 1991;87:9-16; Wilson, et al . : Vitamin E. antioxidants and lipid peroxidation in experimental atherosclerosis of rabbits. J. Nutr 1978;108:1858-1867; Smith, et al . : Effect of dietary vitamin E on plasma lipids and atherogenesis in restricted ovulator chickens. Atherosclerosis 1989;75:105-109; Hayashi . et al.
  • MDA malondialdehyde
  • Macrophage- derived foam cells freshly isolated from rabbit atherosclerotic lesions degrade modified lipoproteins, promote oxidation of low-density lipoproteins, and contain oxidation-specific lipid-protein adducts. J Clin Invest 1991:87:90-99].
  • the antioxidant probucol inhibits degradation of LDL protein in atherosclerotic lesions [Carew, et al . : Antiatherogenic effect of probucol unrelated to its hypocholesterolemic effect: Evidence that antioxidants in vivo can selectively inhibit low density lipoprotein degradation in macrophage-rich fatty streaks and slow the progression of atherosclerosis in the Watanabe heritable hyperlipidemic rabbit.
  • Oxidized LDL acts as a chemotactic stimulus for monocytes, potentially drawing these cells into arteries, and inhibits the mobility of these cells once they have undergone transition into tissue macrophages so they remain within the artery.
  • Oxidized LDL promotes cholesterol ester accumulation in themacrophages, potentiallyconverting arterial macrophages to lipid filled cells resembling "foam cells", the major constituents of the atherosclerotic lesions known as "fatty streaks" [Parthasarathy, et al.: The role of oxidized low-density lipoprotein in the pathogenesis of atherosclerosis, Annu. Rev. Med.1992;43:219-225]. Beta migrating very low density lipoprotein ( ⁇ -VLDL), a cholesterol-enriched VLDL found in plasma of cholesterol fed rabbits also can be oxidatively modified and in this form also promotes cholesterol accumulation in macrophages [Parthasarathy.
  • ⁇ -VLDL very low density lipoprotein
  • Oxidative modification of beta-very low density lipoprotein Potential role in monocyte recruitment . and foam cell formation. Arteriosclerosis 1989;9:398-404].
  • intra-arterial oxidative modification of LDL or ⁇ - VLDL could provide a mechanism for recruiting monocytes into the artery and for conversion of the resulting tissue macrophages into foam cells.
  • Oxidatively modified LDL could promote atherogenesis in other ways, including influencing the production of adhesion ' molecules and cytokines by arterial cells, causing cellular death, and serving as a chemoattractant for smooth muscle cells [Parthasarathy, et al .
  • rabbits were either fed a diet that is known to cause atherosclerosic lesions in rabbits (atherogenic diet, control group) or the same diet supplemented with moderate levels of the 3 antioxidant nutrients.
  • the hypothesis that these nutrients inhibit atherosclerosis by an antioxidant mechanism was investigated by isolating LDL and /?-VLDL from the plasma of these rabbits after supplementation and assessing the susceptibility of these lipoproteins to in vitro oxidation.
  • the control diet contained enough vitamin E to prevent deficiency.
  • the diets containing combinations of antioxidant nutrients were formulated from information synthesized from previous in vivo experiments in rabbits described above.
  • the study included a group of animals fed the atherogenic diet supplemented with probucol, a lipid-lowering antioxidant drug that has been found in previous work to reduce fatty streak formation.
  • the mean body weights for each group were 2.5 ⁇ to 0.1 kg.
  • the first group of rabbits was fed an atherogenic diet containing 10.7* butter and an added 928 mg cholesterol per kg (see Table 1).
  • a second group of rabbits was fed the same atherogenic diet supplemented with vitamin E (2660 IU in the form of ⁇ -tocopherol acetate per kg diet.54 times the amount of the basal diet).
  • a third group was fed the atherogenic diet supplemented with the same level of vitamin E and selenium (410 ⁇ g of elemental selenium per kg diet, 10 times the level in the basal diet).
  • a forth group of rabbits was fed the atherogenic diet supplemented with vitamin E and vitamin C (14.4 g per kg diet).
  • the fifth group was fed the atherogenic diet supplemented with all 3 antioxidant components at the levels specified.
  • Finally the sixth group was fed the atherogenic diet supplemented with 7.38 g of probucol per kg of diet.
  • Probucol sold under the brand name of Lorelco by Merrell Dow (Cincinnati, Ohio) is the generic- name of 4 ' ,4 ' -(isopropylidene dithio)bis(2.6,-di-tert-butyl)phenol.
  • Vitamin D3 1.45 100,000 IU/gm
  • the daily dose is based on 55 grams of diet fed per day
  • Vitamin E 4.9 IU/lOOg 271 IU/100 g 149 IU
  • Vitamin C None added 1440 mg/100 g 791 mg
  • Atherosclerosis was induced by 22 weeks of treatment with the above described diets. Every 2 weeks during this period, blood samples were collected after an overnight fast for measurement of plasma cholesterol concentrations. Every 4 weeks (alternate blood samples) plasma triglycerides were also measured. Plasma samples were collected every 4 weeks for measurement of vitamin E, vitamin C, and selenium.
  • the HPLC method used to measure vitamin E also provides values of ⁇ -carotene, lycopene and other carotenoids, should they be present.
  • Lipoproteins were isolated at three times: After the animals had become adjusted to the animal facility (5-7 days after receipt) and while the animals were consuming a standard rabbit chow (Prolab ® High Fiber Rabbit Formula; from Agway, Ithaca, New York); after 12 weeks of atherogenic diet; and just prior to sacrifice after 22 weeks of atherogenic diet. Lipoprotein fractions were characterized by the cholesterol and protein contents. Selected lipoprotein fractions (LDL during basal diet, /?-VLDL and LDL at weeks 12 and 22 of treatment) were tested for susceptibility to in vitro lipid peroxidation. Samples of LDL and ?-VLDL were stored at -70°C for future characterization by the concentrations of vitamin E and other antioxidants.
  • Values of antioxidants present in LDL will be used to interpret differences in susceptibility of LDL and ?-VLDL to in vitro lipid peroxidation among the groups.
  • the susceptibility of LDL and ?-VLDL to in vitro lipid peroxidation may well reflect the relative potential for intra- arterial oxidation of these lipoproteins among the different treatments.
  • a blood sample was collected for isolation of lipoproteins and the rabbits were sacrificed with an overdose of sodium pentobarbital (100 mg per kg).
  • the entire aorta from aortic arch to just distal of the iliac bifurcation was removed, freed of adventitial debris, opened longitudinally and pinned flat.
  • the percentage of the aortic surface involved with atherosclerotic lesions was determined visually for each of the aortic arch, descending thoracic aorta, and abdominal aorta. Photographs were taken-for measurement of lesion areas by planimetry using a digitizing pad and appropriate software; these measurements are in progress.
  • aortic arch, descending thoracic aorta and abdominal aorta were frozen at -70°C under nitrogen or argon.
  • Aortic samples will be minced and lipids extracted by the method of Folch. The average "severity" of atherosclerosis in each individual aortic sample will be assessed by measuring the esterified and nonesterified cholesterol content of an aliquot of the lipid extract. Concentrations of Vitamin E and other lipophilic antioxidants, such as ⁇ -carotene and lycopene, that might be present will be measured in another aliquot of the lipid extract.
  • the decreased slope of the relationship between Chol lndex and atherosclerosis in treated rabbits indicates that antioxidants interfere with mechanism(s) (such as intraarterial oxidation of lipoproteins) by which plasma l ipoproteins promote atherosclerosis and that such effects can be increased by combining antioxidants with different site(s) and mechanism(s) of action.
  • mechanism(s) such as intraarterial oxidation of lipoproteins
  • combined treatment with vitamin E and selenium inhibited atherosclerosis as effectively as probucol .
  • Int. J . Vita Nutr. Res . 1976; 46:275-285] (measures of severity of atherosclerotic lesions) can be reduced by vitamin C treatment without effect on surface area of aorta involved with atherosclerotic lesions [Hayashi , et al . : Fundamental studies on physiological and pharmacological actions of L-ascorbate 2-sulfate. V. On the hypolipidemic and anti atherosclerotic effects of L-ascorbate 2-sulfate in rabbits .
  • Values are means ⁇ SEM for the indicated number of animals.
  • vitamin E was added to the vitamin E fortified diets at 54 times the National Research Council requirement for rabbits.
  • the same method can be used to calculate a minimum dose for humans: assuming that 5 mg/day of ⁇ -tocopherol acetate is a minimum amount required to prevent deficiency in humans [National Research Council: Recommended Dietary Allowances, ed 10., 1989; pll5-124] and that an intake of approximately 50 times that level would be effective, the protective effect of this antioxidant nutrient would occur at 250 mg per day or 250 IU per day of ⁇ -tocopherol acetate or 186.6 mg per day or 250 IU per day of RRR ⁇ -tocopherol .
  • the upper level of vitamin E intake can be calculated to provide an equivalent dose of vitamin E on a weight/weight basis for an 80 kg human subject as was provided for a 3 kg rabbit: 146 IU per day for a 3 kg rabbit is equivalent to a dose of 3893 IU per day for an 80 kg human subject, or approximate by 4000 IU per day.
  • 146 IU per day for a 3 kg rabbit is equivalent to a dose of 3893 IU per day for an 80 kg human subject, or approximate by 4000 IU per day.
  • Ranges of vitamin E that have been effective in improving cardiovascular disease (e.g. intermittent claudication) when administered alone are within the above range, namely between 400 and 1600 IU per day as reported by [Kleijnen, et al . : Vitamin E and Cardiovascular Disease in the EUR J Clin Pharmacol ,1989:37:541-544].
  • vitamin C intake for protection against atherosclerosis in humans is confounded by the fact that rabbits, unlike humans, do not have an absolute requirement for vitamin C. Therefore, the only reasonable approach is to recommend vitamin C intakes that exceed the RDA by at least two-fold and up to a maximum level of fifty-fold the RDA, e.g. levels of 120 mg per day to 3,000 mg per day.
  • a narrower range of vitamin C intake combined with vitamin E and selenium is between 10 and 20 times the RDA for adult males, that is between 600 and 1,200 mg/day.
  • the studies described above are directed at the hypothesis that the nutrient antioxidants vitamin E, selenium, and vitamin C in combination and in the range of concentrations listed above can inhibit atherosclerosis by inhibiting oxidation of LDL.
  • the results suggest that food components not previous recommended for persons with atherosclerosis but containing the proper level of the above antioxidants might provide some protection from the development of atherosclerosis when consumed at safe levels as part of a well balanced diet.
  • the above results also suggest that this combination of antioxidant nutrients can inhibit atherosclerosis to a similar degree as probucol. thus obviating the need for drug treatment in individuals with elevated LDL or VLDL levels who are at risk for atherosclerosis.

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Abstract

Une composition antioxydante entérale nutritive contient de la vitamine E, du sélénium et éventuellement de la vitamine C. Cette composition antioxydante est utile pour inhiber la surface d'athérosclérose dans une artère; réduire l'oxydabilité de la lipoprotéine de faible densité et de très faible densité. Lorsqu'elle contient de la vitamine C, cette composition antioxydante présente une utilité supplémentaire pour inhiber l'épaisseur des lésions athérosclérotiques et le dépôt de cholestérol dans ces dernières.
PCT/US1995/002658 1994-05-02 1995-03-06 Inhibition de l'atherosclerose au moyen d'antioxydants WO1995029684A1 (fr)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6080788A (en) * 1997-03-27 2000-06-27 Sole; Michael J. Composition for improvement of cellular nutrition and mitochondrial energetics
WO2007005670A2 (fr) * 2005-06-30 2007-01-11 The Research Foundation Of The State University Of New York Analogues de soufre et de selenium naturels et synthetiques, et leurs formes conjuguees polymeres, pour la modulation de l'angiogenese
WO2006070022A3 (fr) * 2004-12-31 2007-04-19 Biosyn Arzneimittel Gmbh Medicaments contenant du selenium pour prevenir ou traiter des affections vasculaires endotheliales

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1983001196A1 (fr) * 1981-10-06 1983-04-14 Erland Johansson Melange d'ingestion par l'homme de selenium en tant que substance a l'etat de trace et son utilisation dans des solutions de stockage d'organes, dans des milieux de culture de cellules et dans des solutions nutritives de stockage de composants sanguins
CA2057463A1 (fr) * 1991-12-11 1993-06-12 David Rowland Composition vitaminique/minerale

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1983001196A1 (fr) * 1981-10-06 1983-04-14 Erland Johansson Melange d'ingestion par l'homme de selenium en tant que substance a l'etat de trace et son utilisation dans des solutions de stockage d'organes, dans des milieux de culture de cellules et dans des solutions nutritives de stockage de composants sanguins
CA2057463A1 (fr) * 1991-12-11 1993-06-12 David Rowland Composition vitaminique/minerale

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
J.A.SIMON: "Vitamin C and Cardiovascular Disease: A review", JOURNAL OF THE AMERICAN COLLEGE OF NUTRITION, vol. 11, (US), pages 107 - 125 *
J.WOJCICKI ET AL.: "Effect of selenium and vitamin E on the development of experimental atherosclerosis in rabbits", ATHEROSCLEROSIS, vol. 87, (IE), pages 9 - 16 *
K.F.GEY ET AL.: "Inverse correlation between plasma vitamin E and mortality from ischemic heart disease in cross-cultural epidemiology", AM.J.CLIN.NUTR., vol. 53, (US), pages 326 - 334 *
K.NYYSSÖNEN ET AL.: "Increase in oxidation resistance of atherogenic serum lipoproteins following antioxidant supplementation: a randomized double blind placebo-controlled clinical trial", EUROPEAN JOURNAL OF CLINICAL NUTRITION, vol. 48, no. 9, (UK), pages 633 - 642 *
KEIZO SATO ET AL.: "Free Radical-Mediated Chain Oxidation of Low Density Lipoprotein and Its Synergistic Inhibition by Vitamin E and Vitamin C", ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, vol. 279, no. 2, pages 402 - 405 *
LIDIA ROZEWICKA ET AL.: "Protective effect of selenium and vitamin E against changes induced in heart vessels of rabbits fed chronically on a high-fat diet", KITASATO ARCH. OF EXP.MED., vol. 64, no. 4, (JP), pages 183 - 192 *
T.M.A.BOCAN ET AL.: "Antiatherosclerotic Effects of Antioxidants Are Lesion-Specific When evaluated in Hypercholesteremic New Zealand White Rabbits", EXPERIMENTAL AND MOLECULAR PATHOLOGY, vol. 57, pages 70 - 83 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6080788A (en) * 1997-03-27 2000-06-27 Sole; Michael J. Composition for improvement of cellular nutrition and mitochondrial energetics
WO2006070022A3 (fr) * 2004-12-31 2007-04-19 Biosyn Arzneimittel Gmbh Medicaments contenant du selenium pour prevenir ou traiter des affections vasculaires endotheliales
WO2007005670A2 (fr) * 2005-06-30 2007-01-11 The Research Foundation Of The State University Of New York Analogues de soufre et de selenium naturels et synthetiques, et leurs formes conjuguees polymeres, pour la modulation de l'angiogenese
WO2007005670A3 (fr) * 2005-06-30 2007-07-12 Univ New York State Res Found Analogues de soufre et de selenium naturels et synthetiques, et leurs formes conjuguees polymeres, pour la modulation de l'angiogenese

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