JP2008513350A - Composition comprising flavonoid and tocotrienol and method thereof - Google Patents

Abstract

  In certain embodiments, comprising a pharmaceutical ingredient comprising an active agent combination comprising a flavonoid and tocotrienol in a ratio of about 75:25 to about 95: 5 and at least one pharmaceutically acceptable excipient. Pharmaceutical formulations such as tablets, gel capsules, solutions, solutions, suspensions or emulsions are disclosed. The flavonoid can wrap naringenin, hesperetin, nobiletin or tangerine. The tocotrienol can wrap α-tocotrienol, γ-tocotrienol or δ-tocotrienol. The formulation can be used to reduce total cholesterol, triacylglycerol, LDL cholesterol and Apo B.

Description

  The present invention relates to compositions and methods for preventing and treating cardiovascular diseases such as hypercholesterolemia and atherosclerosis using a combination of flavonoids and tocotrienols.

  Hyperlipidemia is a pathological condition in mammals, and the concentration of circulating lipids in serum is abnormally high. The composition of the circulating lipid pool consists mostly of triglycerides (fatty acid esters of glycerol), cholesterol, and fatty acid esters of cholesterol. These molecules are generally found bound to specific proteins in the form of a complex that acts as a transport mechanism. Hyperlipidemia is a condition that is generally accompanied by increased levels of cholesterol, phospholipids and / or triglycerides in the serum of mammals.

  Hyperlipidemia includes six types of hereditary hyperlipoproteinemia. These types are often referred to as lipoprotein phenotypes. The main plasma lipids (which include cholesterol and triglycerides) do not dissolve and freely circulate in the plasma, but they bind to proteins and are macromolecular complexes called lipoproteins Transported as a body. The classification of inherited hyperlipoproteinemia by phenotype is important because dietary management and drug therapy depend heavily on that information (The Merck Manual, 16.sup.th edition, Robert Berkow and Andrew J. Fletcher, Merck & Co., Inc., Rahway, NJ 1992). The goal in current hyperlipidemia treatment is to lower lipid levels through weight management and dietary management. In addition to diet and weight management, blood lipid lowering drugs (including, for example, indications required) may also be administered.

  Plasma lipoproteins are carriers of lipids from the site where they are synthesized and absorbed to the site where they are stored and / or utilized. Lipoproteins are spherical particles that have triglycerides and cholesterol esters in their cores and layers of phospholipids, non-esterified cholesterol and apolipoproteins on their surfaces. Lipoproteins, based on their hydrated density, are very large particles rich in triglycerides (known as kilomicrons), very low density lipoprotein (VLDL), intermediate density lipoprotein (IDL ), Low density lipoprotein (LDL) and high density lipoprotein (HDL).

  Apolipoprotein is a protein component of lipoprotein that has three main functions, including the following: (1) maintaining the stability of lipoprotein particles; 2) To act as a coenzyme for an enzyme that acts on lipoproteins, and (3) To remove lipoproteins from the circulation by a receptor-mediated mechanism. The four groups of apolipoproteins are apolipoprotein A (Apo A), apolipoprotein B (Apo B), apolipoprotein C (Apo C) and apolipoprotein E (Apo E).

  LDL consists of a hydrophobic lipid core composed of cholesterol esters and triglycerides. The lipid core of LDL particles is surrounded by an amphiphilic layer composed of phospholipids, non-esterified cholesterol and Apo B.

  Several studies have shown that elevated levels of Apo B in the blood are reliable markers for coronary atherosclerosis (Sniderman, A. et al., Proc. Natl Acad. Sci. USA, 77: 604-608 (1980); Kwiterovich, PO et al., Am. J. Cardiol., 71: 631-639 (1993); McGill et al. Coron. Artery Dis., 4 : 261-270 (1993); Tornvall, P. et al., Circulation, 88: 2180-2189 (1993)).

  In the United States, complications of arteriosclerosis are the leading cause of about half of all deaths and about one third of deaths between the ages of 35 and 65 years. Atherosclerosis or the production of atherosclerotic plaques in the aorta and middle artery is the most common form of arteriosclerosis. Many factors such as age, elevated plasma cholesterol levels, high arterial pressure, smoking, high-density lipophilicity can be used to promote atherosclerosis, regardless of the underlying primary pathogenic change. Lower levels of protein (HDL) cholesterol or family history of premature coronary artery disease are involved.

  The risk of death from coronary artery disease is continuously and stepwise related to total serum cholesterol levels above 180 mg / dL (Stamler, J. et al., (1986) JAMA 256: 2823). About one-third of adults in the United States have levels above 240 mg / dL, and thus have twice the risk of coronary artery disease as humans with cholesterol levels below 180 mg / dL. The promotion of atherosclerosis is primarily correlated with an increase in LDL, or the β fraction is negatively correlated with atherosclerosis (Castelli, WP et al. (1986 ) JAMA 256: 2835). HDL shows a protective effect, and the ratio of total cholesterol to HDL cholesterol is a predictor of coronary artery disease, which is superior to the level of either of them alone. Total cholesterol levels are classified as follows: desirable (<200 mg / dL), borderline high (200-239 mg / dL), or high (> 240 mg / dL) (Report of the National Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (1988) Arch Intern Med 148: 36).

  Advances in research on cholesterol metabolism and coronary disease have started an era of increasing importance on preventive therapy. New guidelines for detecting and treating high blood cholesterol levels in adults indicate that patients with high cholesterol levels or patients whose cholesterol levels are in the border and that have two or more other risk factors are considered LDL Is recommended to be measured. LDL cholesterol levels are classified as follows: borderline risk (130-159 mg / dL) or high risk (> 160 mg / dL). Diet therapy is recommended for patients with high risk levels of LDL and those with borderline risk levels and two or more other risk factors. The Pharmacotherapy for all patients with LDL levels above 189 mg / dL and patients with LDL cholesterol levels between 159 and 189 mg / dL and having two or more other risk factors Is recommended.

  In view of the above, it is not surprising to know that many compounds have been proposed for treating hyperlipidemia in mammals. For example, colestipol hydrochloride (US Pat. No. 3,692,895 and US Pat. No. 3,803,237) is a basic anion exchange resin that sequesters bile acids in the intestine when ingested. This stimulates the production of bile acids, which are greatly reduced using cholesterol stored in the body. This reduces the LDL level. Gemfibrozil (which is described in US Pat. No. 3,674,836) has also been used in the treatment. Niacin (3-pyridinecarboxylic acid) is also administered orally for hypercholesterolemia at a dose of about 1.5-6 g per day. Other medications that are sometimes administered for hyperlipidemia include neomycin, norethindrone acetate, oxandrolone, and dextrothyroxine (Remington's Pharmaceutical Sciences, (17th Ed., Mack Pub. Co., 1985). , pp. 863-865). US Pat. No. 4,499,303 describes the use of a group of N-benzoylsulfamates and benzoylsulfonamides as useful antihyperlipidemic agents. US Pat. No. 4,395,417 proposes the use of cyclic imides, diones, reduced diones and analogs as useful agents.

  The present invention relates to compositions and methods for preventing and treating cardiovascular diseases such as hypercholesterolemia and atherosclerosis using a combination of flavonoids and tocotrienols. Flavonoids are polyphenolic compounds that are ubiquitously present in plant foods (especially orange, grapefruit and tangerine). Tocotrienol is a form of vitamin E present in coconut oil and having unsaturated side chains. Flavonoids and tocotrienols are used to inhibit the production of cholesterol, low density lipoprotein (LDL) and Apo B protein in the prevention and / or treatment of atherosclerosis and / or hypercholesterolemia . Compositions containing citrus flavonoids and tocotrienols are used to prevent and / or inhibit the production of total serum cholesterol, LDL and Apo B.

Epidemiological studies have shown that flavonoids present in flavonoid Mediterranean dishes can reduce the risk of death from coronary heart disease (Hertog, MG et al., 1993, Lancet: 342 , 1007-1011). Soy isoflavones (eg genistein), a minor component of soy protein preparations, can have the effect of lowering cholesterol (Kurowska, EM et al., 1990, J. Nutr. 120: 831-836). Flavonoids present in the juices of citrus fruits such as orange and grapefruit include, but are not limited to, hesperetin and naringenin, respectively. Flavonoids present in tangerine include, but are not limited to, tangerine or nobiletin.

  Tocotrienol is a form of vitamin E present in coconut oil and having unsaturated side chains. Tocotrienols include, but are not limited to, α-tocotrienol, γ-tocotrienol or δ-tocotrienol.

  There is a further need in the art for a mixture for preventing and treating cardiovascular disease comprising flavonoids and tocotrienols.

  Throughout this application, various patents and publications are referred to. The disclosures of these publications and patents are incorporated herein by reference in their entirety.

  The object of the present invention is to provide a pharmaceutical ingredient and / or formulation comprising flavonoids and tocotrienol for the treatment and / or prevention of cardiovascular diseases.

  Yet another object of the present invention is to provide a method for treating cardiovascular diseases by administering pharmaceutical ingredients and / or formulations comprising flavonoids and tocotrienols.

  Another object of the present invention is to provide ingredients / formulations and methods for treating atherosclerosis or hypercholesterolemia by lowering total cholesterol using flavonoids and tocotrienols.

  Another object of the present invention is to provide ingredients / formulations and methods for treating atherosclerosis or hypercholesterolemia by reducing triacylglycerols using flavonoids and tocotrienols.

  Another object of the present invention is to provide ingredients / formulations and methods for treating atherosclerosis or hypercholesterolemia by lowering LDL cholesterol using flavonoids and tocotrienols.

  Another object of the present invention is to provide ingredients / formulations and methods for treating atherosclerosis or hypercholesterolemia by lowering Apo B using flavonoids and tocotrienols.

  Another object of the present invention is to provide ingredients / formulations for treating atherosclerosis or hypercholesterolemia using flavonoids and tocotrienols, where the ingredients / formulations contain low levels of synephrine. And providing a method.

  Some of the above objects of the present invention can be achieved by the present invention. The present invention relates, in certain embodiments, to a pharmaceutical ingredient comprising an active agent combination comprising a polymethoxylated flavonoid and tocotrienol in a ratio of about 75:25 to about 95: 5, wherein the pharmaceutical ingredient comprises From the group consisting of essence oil isolated from citrus fruit, pericarp oil isolated from citrus fruit, pericarp isolated from citrus fruit, decharacterized citrus fruit and combinations thereof Selected.

  In certain embodiments, the invention provides a pharmaceutical ingredient comprising an active agent combination comprising a flavonoid and tocotrienol in a ratio of about 75:25 to about 95: 5 and at least one pharmaceutically acceptable excipient. It relates to a pharmaceutical preparation containing

  In certain embodiments, the present invention relates to a pharmaceutical formulation comprising an active agent combination comprising a flavonoid and a tocotrienol, and a pharmaceutical formulation comprising at least one pharmaceutically acceptable excipient, wherein The formulation reduces total cholesterol by at least 10% after administration.

  In certain embodiments, the present invention relates to a pharmaceutical formulation comprising an active agent combination comprising a flavonoid and a tocotrienol, and a pharmaceutical formulation comprising at least one pharmaceutically acceptable excipient, wherein The formulation reduces triacylglycerol by at least 15% after administration.

  In certain embodiments, the present invention relates to a pharmaceutical formulation comprising an active agent combination comprising a flavonoid and a tocotrienol, and a pharmaceutical formulation comprising at least one pharmaceutically acceptable excipient, wherein The formulation reduces LDL cholesterol by at least 10% after administration.

  In certain embodiments, the present invention relates to a pharmaceutical formulation comprising an active agent combination comprising a flavonoid and a tocotrienol, and a pharmaceutical formulation comprising at least one pharmaceutically acceptable excipient, wherein The formulation reduces Apo B by at least 10% after administration.

  In certain embodiments, the present invention relates to methods for treating cardiovascular disease by administering the components / formulations disclosed herein.

  The term “essence oil” refers to the oil-soluble component (eg, fraction) remaining after the juice has evaporated.

  The term “peeled oil” refers to oil isolated from the peel of citrus fruits.

  The term “pericarp” refers to the pericarp of a citrus fruit, where for the purposes of the present invention the fruit is, for example, dried or chopped. Or may be pelletized.

  The term `` citrus fruit '' refers to the fruit of the genus Citrus, where orange fruit, lemon, lime, tangerine, grapefruit (e.g., pink grapefruit, red peel) Mention may be made of peel) grapefruit) and, in particular, citrus arurentium.

  The term “decharacterized fruit” refers to the fruit after the juice has been extracted. The de-characterized fruit can be, for example, in the form of a mush or mash. The term `` Tomah presscake '' refers to the particularly preferred presscake described in U.S. Pat.No. 5,320,861 and U.S. Pat.No. 5,320,861, present in a presscake made in a conventional manner. Contains higher levels of desirable phytochemicals than are present. In particular, de-characterized cranberry fruits in the form of “Thoma Press Cake” contain higher levels of anthocyanins, phenolic acids and proanthocyanidins than are found in press cakes made by conventional methods. Yes. For example, the anthocyanin content is typically 30% or more of the anthocyanins present in natural cranberry fruits, and the phenolic acid content is typically phenol present in natural cranberry fruits. More than 8% of the acid and the proanthocyanidin content is typically more than 60% of the proanthocyanidins present in natural cranberry fruits.

  The term “isolated” refers to the removal or change of the composition or compound from its native state.

  The term “flavonoid” includes, but is not limited to, polymethoxylated flavonoids. The term “flavonoid” refers to any member of the group of aromatic, oxygen-containing, heterocyclic pigments found in the derivatives of the present invention, including, for example, members of the following chemical subgroups: ) Catechins, (2) leucoanthocyanidins and flavanones, (3) flavanins, flavones and anthocyanins, and (4) flavonols. In a preferred embodiment, the flavonoid includes, for example, proanthocyanidins, flavan-3-ols, anthocyanins or flavanols. Examples of such flavonoids include naringenin, hesperetin, nobiletin and / or tangeretin.

  The term `` tocotrienol '' refers to any tocopherol (T) compound or tocotrienol (T3) compound, such as α-tocopherol, γ-tocopherol, δ-, present at a measurable level in the fruit derivatives of the present invention. Tocopherol, α-tocotrienol, γ-tocotrienol, δ-tocotrienol or combinations thereof are indicated.

  The term “pharmaceutical component” means a therapeutic composition capable of providing a pharmaceutical formulation or dosage form, optionally in combination with a pharmaceutically acceptable excipient.

  The term “pharmaceutical formulation” means a pharmaceutical ingredient combined with at least one pharmaceutically acceptable excipient. The formulation can be administered in any acceptable form (eg, tablet or capsule) by any acceptable route (eg, oral route).

  In certain embodiments, the invention relates to a pharmaceutical ingredient comprising an active agent combination comprising a polymethoxylated flavonoid and tocotrienol in a ratio of about 75:25 to about 95: 5, wherein the pharmaceutical ingredient comprises Selected from the group consisting of essence oil isolated from citrus fruit, peel oil isolated from citrus fruit, peel isolated from citrus fruit, citrus fruit de-characterized and combinations thereof .

  In certain embodiments, the active agent combination comprises flavonoids and tocotrienols in the following ratios: a ratio of about 90:10; a ratio of about 80:20; or a ratio of about 95: 5.

  In certain embodiments, the pharmaceutical ingredients of the present invention comprise about 50% to about 90% flavonoids and tocotrienols; about 60% to about 80% of the active agent combination; or about 70% of the active agent combination. It is out.

  In certain embodiments, the pharmaceutical ingredient contains less than about 1% synephrine; less than about 0.5% synephrine; or less than 0.1% synephrine.

  In certain embodiments, the formulation contains less than about 1% synephrine; less than about 0.5% synephrine; or less than 0.1% synephrine.

  The flavonoids of the present invention can be polymethoxylated flavonoids. In certain embodiments, the flavonoid comprises a member selected from the group consisting of naringenin, hesperetin, nobiletin, tangerine and combinations thereof.

  The tocotrienol of the present invention can be selected, for example, from the group consisting of α-tocotrienol, γ-tocotrienol, δ-tocotrienol and combinations thereof.

  In certain embodiments, the invention provides a pharmaceutical ingredient comprising an active agent combination comprising a flavonoid and tocotrienol in a ratio of about 75:25 to about 95: 5 and at least one pharmaceutically acceptable excipient. It relates to a pharmaceutical preparation containing

  In certain embodiments, the pharmaceutical formulation of the present invention comprises an essence oil isolated from citrus fruit, a peel oil isolated from citrus fruit, a peel isolated from citrus fruit, a decharacterized citrus fruit And a pharmaceutical ingredient selected from the group consisting of combinations thereof.

  In certain embodiments, the pharmaceutical component of a formulation of the invention is at risk for cardiovascular disease (eg, hypercholesterolemia or atherosclerosis) or cardiovascular disease (eg, hypercholesterolemia) Or an amount effective to treat a human subject suffering from atherosclerosis).

  In certain embodiments, the pharmaceutical formulations of the invention are suitable for intravenous, intraperitoneal, subcutaneous, intramuscular, intrathecal, oral, rectal, topical or inhalation administration .

  In certain embodiments, the pharmaceutical formulations of the invention are in the form of tablets, capsules, solutions, solutions, suspensions or emulsions.

  In certain embodiments, the pharmaceutical formulation of the present invention contains about 60 mg tocotrienol and about 560 mg flavonoid per unit dose; about 10 mg to about 80 mg tocotrienol and about 150 mg to about 750 mg flavonoid per unit dose Or about 30 mg tocotrienol and about 270 mg flavonoids per unit dose.

  In the methods of the invention, the daily dose of the active agent is, for example, about 60 mg tocotrienol and about 560 mg flavonoid; about 10 mg to about 80 mg tocotrienol and about 150 mg to about 750 mg flavonoid; or about 30 mg tocotrienol and It can be about 270 mg of flavonoid.

  In the methods of the invention, the flavonoid and the tocotrienol can be administered in the same dosage form, or can be administered in separate dosage forms. Further, the flavonoid and the tocotrienol can be administered by the same route of administration or can be administered by different routes of administration.

  In certain embodiments, the pharmaceutical formulations of the invention reduce total cholesterol by at least 10%, 20% or 30%.

  In certain embodiments, the pharmaceutical formulations of the invention are administered after a single dose administration or after multiple dose administration (e.g. after 4 weeks) or after steady state administration. In a single patient or in a patient population, total cholesterol is reduced by at least 10%, 20% or 30%.

  In certain embodiments, the pharmaceutical formulations of the invention reduce triacylglycerol by at least 15%, 25% or 35%.

  In certain embodiments, the pharmaceutical formulations of the invention are administered in one patient or after a single dose or after repeated administration (eg, after 4 weeks), or after steady state administration, or patients In the population, triacylglycerol is reduced by at least 15%, 25% or 35%.

  In certain embodiments, the pharmaceutical formulations of the invention reduce LDL cholesterol by at least 10%, 20% or 30%.

  In certain embodiments, the pharmaceutical formulations of the invention are administered in one patient or after a single dose or after repeated administration (eg, after 4 weeks), or after steady state administration, or patients In the population, LDL cholesterol is reduced by at least 10%, 20% or 30%.

  In certain embodiments, the pharmaceutical formulations of the invention reduce Apo B by at least 10%, 20% or 30%.

  In certain embodiments, the pharmaceutical formulation of the present invention provides total cholesterol in a single patient or in a patient population after single or repeated administration (eg, after 4 weeks) or after steady state administration. Is reduced by at least 10%, 20% or 30%.

  In certain embodiments, a pharmaceutical formulation of the invention comprises an active agent combination comprising a flavonoid and a tocotrienol, and further comprises soy protein, soy isoflavone, grape seed extract, pine bark extract, gugulipid, polycosinol an additional active agent selected from the group consisting of (policosinol), pantesine, niacin, α-lipoic acid, tea flavin, coenzyme Q10, lutein, statin and combinations thereof.

  In certain embodiments, the statin drug is selected from the group consisting of pravastatin, simvastatin, lovastatin, atorvastatin, fluvastatin, cerivastatin, and combinations thereof.

  The pharmaceutical formulations of the present invention can be prepared for oral administration, sublingual administration, inhalation administration, subcutaneous administration, intramuscular administration, intravenous administration or transdermal administration, and as a formulation for topical administration or rectal administration. it can. Oral preparations can be, for example, forms such as tablets, gel capsules, powders and granules, and oral solutions or suspensions, sublingual and oral dosage forms.

  When preparing solid compositions in the form of tablets or gel capsules, the active ingredient is micronized or non-micronized with diluents (e.g. lactose, microcrystalline cellulose, starch, dicalcium phosphate). ), Binders (e.g. polyvinylpyrrolidone, hydroxypropylmethylcellulose), crumbling agents (e.g. crosslinked polyvinylpyrrolidone, crosslinked carboxymethylcellulose), flow agents (e.g. silica or talc) and lubricants. Add a mixture of pharmaceutical excipients that may consist of a excipient (eg, magnesium stearate, stearic acid, glyceryl tribehenate or sodium stearyl fumarate).

  Wetting agents or surfactants such as sodium lauryl sulfate, polysorbate 80 or poloxamer 188 can be added to the formulation.

  The tablets can be prepared by various techniques (direct tableting, dry granulation, wet granulation, hot melt).

  The tablets may be uncoated, coated (e.g., with sucrose), or coated with various polymers (e.g., hydroxypropyl methylcellulose) or another suitable material. Good.

  The tablets can be immediate release, delayed release or sustained release by preparing a matrix or using a coating.

  The gel capsules can be soft or hard, and can be coated with a film or otherwise used to exhibit immediate or sustained or delayed activity. (Eg enteric form).

  Oral preparations can also be prepared as liquid or semi-solid preparations (e.g., preparations in the form of syrups or elixirs), as sweeteners (preferably non-caloric sweeteners), and as preservatives. The active ingredients can be included together with methyl and propylparabens of the present and flavoring and suitable colorants.

  A water-dispersible powder or granule can contain the active ingredient as a mixture of a dispersant, a wetting agent or a suspending agent (eg, polyvinylpyrrolidone) and a sweetener or flavor enhancer.

  For rectal administration, suppositories prepared with binders that melt at rectal temperature (eg, cocoa butter or polyethylene glycol) are used.

  For parenteral or intranasal administration, aqueous suspensions containing pharmacologically compatible dispersants and / or solubilizers (eg, propylene glycol), isotonic saline solutions or sterile injectable Use the solution.

  Thus, to prepare an intravenously injectable aqueous solution, a cosolvent such as an alcohol (e.g., ethanol) or glycol (e.g., polyethylene glycol or propylene glycol) and a hydrophilic surfactant (e.g., polysorbate 80 or poloxamer 188) ) Can be used. In order to prepare an injectable oily solution for intramuscular administration, the active ingredient can be dissolved using a triglyceride or glycerol ester.

  For topical administration, creams, ointments, gels, transdermal patches and sprays can be used. For transdermal administration, a patch in the form of a reservoir or a multilayer form in which the spray and the active ingredient can be an alcohol solution can be used.

  For administration by inhalation, aerosols such as sorbitan trioleate or oleic acid and trichlorofluoromethane, dichlorofluoromethane, dichlorotetrafluoroethane, freon substitutes or any other biocompatible jet Use aerosol containing agent gas. It is also possible to use a system in powder form containing the active ingredient alone or containing the active ingredient in combination with excipients.

  The active ingredient can also be formulated in the form of microcapsules or microspheres, optionally with one or more supports or additives.

  Of the sustained release forms useful for long-term treatment, implants can be used. They can be prepared in the form of an oily suspension or in the form of a suspension of microspheres in an isotonic medium.

Example 1
We tested the effect of the combination of flavone and tocotrienol on lowering cholesterol in human subjects.

Test design:
Ten subjects with hypercholesterolemia (serum total cholesterol> 5.9 mmol / L, LDL cholesterol 4.0 mmol / L, and serum triacylglycerol <3.5 mmol / L (> 230 mg / dL,> 155 mg / L, respectively) dL and <307 mg / dL)) were given daily supplements consisting of 270 mg polymethoxyflavone and 30 mg tocotrienol for 4 weeks. In order to participate in this study, it was essential that the subject did not suffer from thyroid disease, kidney injury and diabetes. Furthermore, subjects taking cholesterol-lowering drugs were required to discontinue their treatment for 4 weeks prior to the start of the study.

  To determine if this treatment has improved parameters associated with high risk of heart disease, plasma total cholesterol and plasma lipoprotein cholesterol, plasma apolipoprotein B (LDL and LDL) at the start of the study and at the end of 4 weeks. Fasting blood samples for analysis for (related) and plasma apolipoprotein A1 (related to HDL) and total triacylglycerol were drawn. This protocol was approved by the University of Western Ontario's Human Ethics Committee and informed consent was obtained from each subject.

  Cholesterol and triacylglycerol were measured by an enzymatic timed-endpoint method by using CHOL reagent or triacylglycerol GPO reagent. Plasma concentrations of Apo B and Apo A1 were analyzed immunoephelometrically using the BNII System.

  Subjects were instructed to maintain their caloric intake throughout the study period. This was evaluated by measuring the body mass index (BMI) before and after treatment.

  Changes from baseline after 4 weeks were analyzed using repeated measures analysis of variance (ANOVA) followed by Dunnett's test.

Result :
Treatment with a combination of 90% flavonoids and 10% tocotrienol had many beneficial effects. Treatment with this combination significantly reduced total cholesterol, LDL cholesterol and serum triacylglycerol. See Table 1. This result suggests that the combination of flavonoids and tocotrienols lowers human cholesterol.

Example 2
Clinical Trial 2: Cholesterol-lowering properties of polymethoxylated flavone and tocotrienol combination in human subjects The purpose of this study is to evaluate the potential of flavone and tocotrienol combination to protect the heart in human subjects Met.

Test design:
Ten subjects with hypercholesterolemia were given a supplement consisting of 270 mg polymethoxyflavone and 30 mg tocotrienol daily for 4 weeks. In order to participate in this study, it was essential that the subject did not suffer from thyroid disease, kidney injury and diabetes. Furthermore, subjects taking cholesterol-lowering drugs were required to discontinue their treatment for 4 weeks prior to the start of the study.

  Blood samples were collected from the forearm veins before the start of the study (baseline) and at the end of the 4 week period. Plasma lipid profiles and other metabolic parameters were analyzed using standard methods. Blood pressure was recorded in a sitting position using a conventional mercury sphygmomanometer and the average of two measurements.

Result :
Treatment with a combination of 90% flavonoids and 10% tocotrienol had many beneficial effects. Treatment with this combination reduced total cholesterol (19.7%), LDL cholesterol (22.01%), serum triacylglycerol (28.4%) and Apo B (20.9%) and decreased systolic blood pressure. See Table 2. This result suggests that the combination of flavonoids and tocotrienol has the potential to protect the heart of subjects suffering from moderate hypercholesterolemia.

Claims (85)

  A pharmaceutical ingredient comprising an active agent combination comprising a polymethoxylated flavonoid and tocotrienol in a ratio of about 75:25 to about 95: 5, wherein the pharmaceutical composition is isolated from citrus fruit Said medicinal ingredient selected from the group consisting of oil, peel oil isolated from citrus fruit, peel isolated from citrus fruit, de-characterized citrus fruit and combinations thereof.   The pharmaceutical ingredient according to claim 1, wherein the active agent combination comprises a flavonoid and tocotrienol in a ratio of about 90:10.   The pharmaceutical ingredient according to claim 1, wherein the active agent combination comprises a flavonoid and tocotrienol in a ratio of about 80:20.   The pharmaceutical ingredient according to claim 1, wherein the active agent combination comprises a flavonoid and tocotrienol in a ratio of about 95: 5.   2. The pharmaceutical ingredient of claim 1 comprising from about 50% to about 90% of the active agent combination.   2. The pharmaceutical ingredient of claim 1 comprising from about 60% to about 80% of the active agent combination.   2. The pharmaceutical ingredient of claim 1 comprising about 70% of the active agent combination.   The pharmaceutical ingredient according to claim 1, wherein the flavonoid comprises a member selected from the group consisting of naringenin, hesperetin, nobiletin and tangeretin.   The pharmaceutical ingredient according to claim 1, wherein the tocotrienol is selected from the group consisting of α-tocotrienol, γ-tocotrienol and δ-tocotrienol.   A pharmaceutical formulation comprising a pharmaceutical ingredient comprising an active agent combination containing flavonoids and tocotrienol in a ratio of about 75:25 to about 95: 5 and at least one pharmaceutically acceptable excipient.   The pharmaceutical ingredient comprises essence oil isolated from citrus fruits, pericarp oil isolated from citrus fruits, pericarp isolated from citrus fruits, de-characterized citrus fruits and combinations thereof 11. A pharmaceutical formulation according to claim 10 selected from the group.   11. The pharmaceutical formulation of claim 10, wherein the pharmaceutical ingredient is present in an amount effective to treat a human subject at risk for or suffering from cardiovascular disease.   The pharmaceutical preparation according to claim 10, wherein the cardiovascular disease is hypercholesterolemia or atherosclerosis.   The pharmaceutical preparation according to claim 10, suitable for intravenous administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intrathecal administration, oral administration, rectal administration, topical administration or administration by inhalation.   The pharmaceutical formulation according to claim 14, which is suitable for intravenous administration.   The pharmaceutical formulation according to claim 14, which is suitable for oral administration.   The pharmaceutical preparation according to claim 10, which is in the form of a tablet, capsule, gel capsule, solution, solution, suspension or emulsion.   11. The pharmaceutical formulation of claim 10, comprising from about 10 mg to about 80 mg of the tocotrienol and from about 150 mg to about 750 mg of the flavonoid per unit dose.   11. The pharmaceutical formulation of claim 10, comprising about 30 mg of the tocotrienol and about 270 mg of the flavonoid per unit dose.   11. The pharmaceutical formulation of claim 10, comprising about 60 mg of the tocotrienol and about 560 mg of the flavonoid per unit dose.   11. A pharmaceutical formulation according to claim 10 which reduces total cholesterol by at least 10%.   22. A pharmaceutical formulation according to claim 21 which reduces total cholesterol by at least 10% in one patient.   23. The pharmaceutical formulation of claim 21, wherein the pharmaceutical formulation reduces total cholesterol by at least 10% in the patient population.   24. A pharmaceutical formulation according to any one of claims 21 to 23 which reduces total cholesterol by at least 10% after a single administration.   24. A pharmaceutical formulation according to any one of claims 21 to 23, which reduces total cholesterol by at least 10% after repeated administration.   24. A pharmaceutical formulation according to any one of claims 21 to 23, which reduces total cholesterol by at least 10% after steady state administration.   26. The pharmaceutical formulation of claim 25, which reduces total cholesterol by at least 10% after 4 weeks of administration.   28. A pharmaceutical formulation according to any one of claims 21 to 27, which reduces total cholesterol by at least 20%.   28. A pharmaceutical formulation according to any one of claims 21 to 27, which reduces total cholesterol by at least 30%.   11. A pharmaceutical formulation according to claim 10 which reduces triacylglycerol by at least 15%.   32. The pharmaceutical formulation of claim 30, wherein the triacylglycerol is reduced by at least 15% in one patient.   32. The pharmaceutical formulation of claim 30, wherein triacylglycerol is reduced by at least 15% in a patient population.   33. A pharmaceutical formulation according to any one of claims 30 to 32, wherein triacylglycerol is reduced by at least 15% after a single administration.   33. A pharmaceutical formulation according to any one of claims 30 to 32, wherein triacylglycerol is reduced by at least 15% after repeated administration.   33. A pharmaceutical formulation according to any one of claims 30 to 32, wherein triacylglycerol is reduced by at least 15% after steady state administration.   35. The pharmaceutical formulation of claim 34, wherein triacylglycerol is reduced by at least 15% after 4 weeks of administration.   37. A pharmaceutical formulation according to any one of claims 30 to 36, wherein triacylglycerol is reduced by at least 25%.   37. A pharmaceutical formulation according to any one of claims 30 to 36, wherein triacylglycerol is reduced by at least 35%.   11. A pharmaceutical formulation according to claim 10 which reduces LDL cholesterol by at least 10%.   40. The pharmaceutical formulation of claim 39, which reduces LDL cholesterol by at least 10% in one patient.   40. The pharmaceutical formulation of claim 39, which reduces LDL cholesterol by at least 10% in a patient population.   42. A pharmaceutical formulation according to any one of claims 39 to 41 which reduces LDL cholesterol by at least 10% after a single administration.   42. A pharmaceutical formulation according to any one of claims 39 to 41 which reduces LDL cholesterol by at least 10% after repeated administration.   42. A pharmaceutical formulation according to any one of claims 39 to 41, which reduces LDL cholesterol by at least 10% after steady state administration.   44. The pharmaceutical formulation of claim 43, which reduces LDL cholesterol by at least 10% after 4 weeks of administration.   46. The pharmaceutical formulation according to any one of claims 39 to 45, which reduces LDL cholesterol by at least 20%.   46. A pharmaceutical formulation according to any one of claims 39 to 45, which reduces LDL cholesterol by at least 30%.   11. A pharmaceutical formulation according to claim 10 which reduces Apo B by at least 10%.   49. The pharmaceutical formulation of claim 48, which reduces Apo B by at least 10% in one patient.   49. The pharmaceutical formulation of claim 48, wherein Apo B is reduced by at least 10% in the patient population.   51. A pharmaceutical formulation according to any one of claims 48 to 50, which reduces Apo B by at least 10% after a single administration.   51. A pharmaceutical formulation according to any one of claims 48 to 50 which reduces Apo B by at least 10% after repeated administration.   51. A pharmaceutical formulation according to any one of claims 48 to 50 which reduces Apo B by at least 10% after steady state administration.   53. The pharmaceutical formulation of claim 52, which reduces Apo B by at least 10% after 4 weeks of administration.   55. A pharmaceutical formulation according to any one of claims 48 to 54 which reduces Apo B by at least 20%.   55. A pharmaceutical formulation according to any one of claims 48 to 54 which reduces Apo B by at least 30%.   A pharmaceutical ingredient comprising an active agent combination containing flavonoids and tocotrienol, and further comprising soy protein, soy isoflavone, grape seed extract, pine bark extract, gugulipid, polycosinol, panthecin, niacin, α-lipoic acid, tea flavin, A pharmaceutical formulation comprising an additional active agent selected from the group consisting of coenzyme Q10, lutein, statin and combinations thereof.   Selected from the group consisting of soy protein, soy isoflavone, grape seed extract, pine bark extract, gugulipid, polycosinol, panthecin, niacin, α-lipoic acid, tea flavin, coenzyme Q10, lutein, statin and combinations thereof 57. The pharmaceutical formulation according to any one of claims 10 to 56, further comprising an additional active agent.   59. A pharmaceutical formulation according to any one of claims 57 to 58, wherein the additional active agent is a statin drug.   60. The pharmaceutical formulation of claim 59, wherein the statin drug is selected from the group consisting of pravastatin, simvastatin, lovastatin, atorvastatin, fluvastatin, cerivastatin and combinations thereof.   21. A method of treating a human subject having a risk of cardiovascular disease or suffering from cardiovascular disease, comprising an effective amount of the pharmaceutical formulation of any one of claims 10-20. Said method comprising administering.   62. The method of claim 61, wherein the cardiovascular disease is hypercholesterolemia or atherosclerosis.   30. A method of reducing a patient's total cholesterol, comprising administering to a patient in need thereof the pharmaceutical formulation of any one of claims 21-29.   39. A method of lowering a patient's triacylglycerol, comprising administering to a patient in need thereof a pharmaceutical formulation according to any one of claims 30-38.   48. A method of lowering LDL cholesterol in a patient comprising administering to a patient in need thereof a pharmaceutical formulation according to any one of claims 39-47.   57. A method of reducing Apo B in a patient, comprising administering to a patient in need thereof a pharmaceutical formulation according to any one of claims 48-56.   67. The method of any one of claims 61-66, wherein the combination is administered at a daily dose of about 10 mg / day to about 80 mg / day tocotrienol and about 150 mg / day to about 750 mg / day flavonoids.   67. The method of any one of claims 61 to 66, wherein the combination is administered at a daily dose of about 30 mg / day tocotrienol and about 270 mg / day flavonoids.   67. The method of any one of claims 61-66, wherein the combination is administered at a daily dose of about 60 mg / day tocotrienol and about 560 mg / day flavonoids.   68. The method of any one of claims 61 to 66, wherein the administration is a single administration.   67. The method according to any one of claims 61 to 66, wherein the administration is repeated administration.   67. The method of any one of claims 61-66, wherein the administration results in a steady state.   72. The method of claim 71, wherein the administration is at least 4 weeks.   64. The method of claim 63, wherein the administration reduces total cholesterol by at least 10%.   65. The method of claim 64, wherein the administration reduces triacylglycerol by at least 15%.   66. The method of claim 65, wherein the administration reduces LDL cholesterol by at least 10%.   68. The method of claim 66, wherein the administration reduces Apo B by at least 10%.   A pharmaceutical formulation comprising a pharmaceutical ingredient comprising an active agent combination comprising flavonoids and tocotrienol and at least one pharmaceutically acceptable excipient, wherein said pharmaceutical formulation reduces total cholesterol by at least 10% after administration.   A pharmaceutical formulation comprising a pharmaceutical ingredient comprising an active agent combination comprising flavonoids and tocotrienol and at least one pharmaceutically acceptable excipient, wherein the pharmaceutical formulation reduces triacylglycerol by at least 15% after administration.   A pharmaceutical formulation comprising a pharmaceutical ingredient comprising an active agent combination comprising flavonoids and tocotrienol and at least one pharmaceutically acceptable excipient, wherein said pharmaceutical formulation reduces LDL cholesterol by at least 10% after administration.   A pharmaceutical formulation comprising a pharmaceutical ingredient comprising an active agent combination comprising a flavonoid and a tocotrienol and at least one pharmaceutically acceptable excipient, wherein the pharmaceutical formulation reduces Apo B by at least 10% after administration.   The ingredient according to claim 1, wherein the citrus fruit is citrus aurentium.   12. The formulation of claim 11, wherein the citrus fruit is citrus aurentium.   2. The component of claim 1 comprising less than about 1% synephrine; less than about 0.5% synephrine; or less than 0.1% synephrine.   2. The formulation of claim 1, comprising less than about 1% synephrine; less than about 0.5% synephrine; or less than 0.1% synephrine.