WO2001070029A1 - Compositions and methods of treating, reducing, and preventing cardiovascular diseases and disorders with polymethoxyflavones - Google Patents

Abstract

Compositions and methods for the treatment, reduction and/or prevention of cardiovascular diseases and disorders are described. Individuals at high risk for developing or having cardiovascular disease or disorder may be treated with an effective dose of a polymethoxyflavone including limocitrin derivatives, quercetin derivatives, naturally occurring polymethoxyflavones, tocotrienols, and mixtures of these compounds.

Description

COMPOSITIONS A D [METHODS OF TREATING, REDUCING, AJND PREVENTING CA-RDIOVASCULA-R DISEASES AJND DISORDERS WITH POLY ETHOXYFLAVONES

1. CROSS REFERENCE TO RELATED APPLICATIONS

This application is a Continuation-in-part of co-pending United Sla.es Patent Application Serial Number 09/167,634, Hied on Oclober 6, 1998 entitled "Compositions and Methods of Inhibiting Neoplaslic Diseases With Compounds Relaled lo Lirnocilrin and 5- Desmelhyl Sineπselin", erein ihe entire disclosure is herein incorporated by A. Field of the Invention reference.

2. BACKGROUND OF THE INVENTION

The present invention relates lo compositions and methods for the prevention, reductiotA, and/or treatment of cardiovascular diseases with synthetic and naturally occurring poiymelhoxyflavone compounds derived, some of which are deiived from limocitiin and quercelin.

These compounds include, but are not limited to, the following examples of lirnocilrin , and queicelin derivatives: liniocilrin-3,7,^'-lrinieth'leiher (5-hydroxy-3,7,S,3',4'-penlamelltoxyπavoπe) limocilrin-3,5,7,4'-lelramelhylether (3,5,7,8,3'4'-he>;amelhoxyflavone) limoci⁽rin-3,5,7₁4'-⁽etraelhylether (S,3'-dimelhoxy-3,5,7,4'-letraelhoxyflavone) lirnocilrin 3,7 -trime(hslethcr-5-acetate quercelin telramethylelher (5-hydroxy -3,7,3', 4'-lelramelhQxyflavone) quercelin 3,5-dimeihylelher-7₁3'4'-lribenzyl ether quercelin pentamcthylether (3,5,7,3',4'-penlamelhoxynavone) quercetiπ-5,7,3', '-ielramelhylelher-3-acelale qιιercelin-5,7,3'₁4'-te(r.ιιnethylclher (3-hydιoNy-5,7,3',T-lelramelhoxyflavone)

Examples, bui not limited lo. of naturally occurring polymethoxyflavones for ihe purposes of the pi esent invention include: 3 ,S,6,7,8,3',4'-heptamcthoxyflavone nobileti (5,6,7,8.3',4'-hcxamethoxyfla\One) tangeretin (5,6,7,8,4'-pen.amethoxyflavone)

5 -desmethy I nobiletin (5-hydroxy-6,7.8.3'4'-pentamethoxyflavone) tetra-O-methylisoscutellarein (S^S^'-tetramethoxyflavone) 5-desmethylsinensctin (5-hydroxy-6,7,3',4'-tetramethoxyflavone) sinensetin (5,6,7,3',4'-pentaιτιethoxyf avone).

B. Description of the Related Art

Limocitrin derivatives are a group of citrus-derived flavonoids that are naturally occurring in the plant or are chemically synthesized. 5-desmethylsinesetin is chemically synthesized form of sinensetin (Tarum. .H. et al.. Phytochemistry II, 2283-2288, 1972). . Sinensetin occurs in trace levels in mandarin orange leaves (Sugiyama, S. et a],. Chem. Pharm. Bull., Volume 4 1 , 7 14-719. 1993), and in orange and mandarin peel. Flavonoids are pσlyphenolic compounds that occur ubiquitously in foods of plant origin. The major dietary sources of flavonoids are \ egetables. fruits, and beverages such as tea and red wine (Hertog, M.G.L. et al., J. Agric. Food Chem.. Volume 4 1 , 1 242- 1246, 1993). Flavonoids have been demonstrated to be the most potent dietary aπtioxidants and in light of the large dietary consumption, flavonoids make a major contribution to the antioxidant potential of the human diet. The main food sources of flavotiols and flavones are black tea, onions, apples, herbs, and spices such as cloves and black pepper (Hertog. M.G.L.. et al, J. Agric. Food Chem., Volume 40, 2379-2383 , 1992).

The association between quercetin and cardiovascular disease has been studied in prospective cohort studies and cross-cultural epidemiological studies. Flavonol and flavone intake was inversely associated with mortality f.om coronary heart disease and to a lesser extent with incidence of first my ocardial infarction. These effects were independent of known risk factors for coronary heart disease such as serum cholesterol, body mass index, blood pressure, smoking and intake of antioxidant vitamins, alcohol, and fat. Flavoπol and flavone intake (mainly quercelin) was also inversely associated with stroke risk (Hertog et al., Lancet, Volume 324, 1007- 1 0 ! I , 1993 ; Kelt et al., Arch. Inter. Med., Volume 1 54, 637-642. 1 996). However, four thousand different types of flavonoids have been described and it is crucial that the active components be identified not only to make a positive impact on agriculture but also to more specifically use these nutraceuticals as anticholesterol agents and/or antitluombotic, anticorσnary heart disease, an imyocardial infarction and/or antistroke agents.

In the United States, the complications of atherosclerosis account for about one half of all deaths and for about one third of deaths in persons between 35 and 65 years of age. Atherosclerosis, or the developments of atheromatous plaques in large and medium-sized arteries, is the most common form of arteriosclerosis. Many factors a re associated with the acceleration of atherosclerosis, regardless of the underlying primary pathogenic change, for example, age, elevated plasma cholesterol level, high arterial blood pressure, cigarette smoking, reduced high-density lipoproteiπ (HDL) cholesterol level, or family history of premature coronary artery disease.

The t isk of death Horn coronary artery disease has a continuous and graded relation to total serum cholesterol levels greater than 1 80 g/dl (Stamler et al,, JAMA, Volume 256, 2823 , 1986). Approximately one third of adults in the United States have levels that exceed 240 mg/dl and, therefore, have a risk of coronary artery disease that is twice that of people with cholesterol levels lower than 180mg/dl. Acceleration of atheiosclevσsis is principally correlated with elevation of LDL. or beta fraction, which is rich in cholesterol but poor in triglycerides, Elevation of HDL or alpha fraction, has a negative correlation with atherosclerosis (Castelli et al,, JAMA, Volume 256, 2835, 1986). HDL exerts a protective effect and the ratio of total cholesterol to HDL cholesterol is a better predictor of coronary artery disease than the level of either alone. Total cholesterol levels are classified as being desirable (<200mg/dl), borderl ine (200-239 mg/dl), or high (>240 mg/dl) (Report of the National Education Expert Panel on Detection. Evaluation, and Treatment of High B lood Cholesterol in Adults, Ai ch Intern. Med.. Volume 148, 36, I 98S ).

Adv ances in the study of cholesterol metabolism and coronary disease have initiated an era of increased emphasis on preventive therapy. New guidelines for the detection and treatment of high blood cholesterol in adults recommend that patients with high cholesterol levels or w ith borderline-high levels and tw o or more additional risk factors should have a measurement of LDL. LDL cholesterol levels are then classified as borderline-high risk ( 130- i 59 mg/dl) or high risk (> 160 mg/dl). Dietary treatment is recommended for those patients with high-risk

els who have two or more additional risk factors. Drug treatment is recommended for all patients with LDL levels greater than 1 S9 mg/dl and for those patients with LDL cholesterol levels between 159 and 189 mg/dl who have tΛvo or more additional risk factors. .Among the many drugs that have been used to reduce serum cholesterol levels are cholestyramiπe, colestipol. clofibrate, gemfibrozil, and lovaslatin.

Platelet-blood vessel interactions are implicated in the

elopment of thrombosis. Flavonoids inhibit platelet aggregation and adhesion (Fran el et al., Lancet, Volu e 34 1 , 1 103- 1 104. 1993). Flavonoids antagonize tliro bσxaπe formation and increase platelet cyclic AMP levels. This is important because flavonoids additional! scavenge free radicals and theii antioxidant actions participate in their aπtithrombotic action (Gryglewski et al.. Biochem. Pharmacol., Volume 36, 3 17-322. 1987). Drug treatment is recommended for patients ith thrombosis and ischemic heart disease. The medical therapy comprises pharmaceutical drugs including, but is not limited to, aspirin (anti-platelet aggregating agents) and the combined use of beta-adrenergic blocking agents (e.g.propranonol, nadσlσl, timolσl, etc.), nitrates (e.g , nitrogh ccrin) and calcium channel blockers (e.g.. verapamil. nifedipine, diltiazem. etc.).

There remains a need in the art for methods and compositions for at least reducing the development of and/or treating vascular diseases. The present inv ention provides new compositions and methods directed to this need. The use of lirnocilrin derivatives, queicetin derivatives and/or, naturally-occurring polymethoxyflavones and mixtures thereof alone or in combination ith a cholesteiol-lowei iπg drug has not been reported for at least reducing the development of and'or ti eating vascular diseases and disorders. 3. SUMMARY OF THE INVENTION

It is therefore an object of the present invention to provide compositions and methods for the reduction, prevention, and/or treatment of cardiovascular diseases and disorders whei ein an effective amount of a composition having at least one limocitrin and/or quercetin derivative is administered to reduce, prevent or treat a mammal at high risk for or suffering from a cardiovascular disease.

Another object of the present invention is to provide compositions and methods for the reduction, prevention, and/or treatment of cardiovascular diseases or disorders wherein an effective amount of a composition having at least one flavonoid is administered to reduce, prevent or treat a mammal at high risk for or suffering from a cardiovascular disease.

A further object of the present invention is to provide compositions and methods for the reduction, prev ention, and/or eatment of cardiovascular diseases or disorders wherein an effective amount of a composition having at least one limocitrin. quercetin derivative, tocotrienol, and mixtures thereof is administered to reduce, prev ent or treat a mammal at high risk for or suffering from a cardiovascular disease.

A still fui ther object of the present invention is to prov tde compositions and methods for the reduction, prevention, and/or treatment of cardiovascular diseases or disorders wherein an effective amount of a composition having al least one naturally occurring polymcthoxyaflavone is administered to reduce, prevent, or treat a mammal at high risk for or suffering from a cardiovascular disease.

Another object of the present invention is to provide compositions and methods for the reduction, prevention, and/or treatment of cardiovascular diseases or disorders wherein an effective amount of a composition having at least one naturally occurring polymethoxyflavone, tocotrienols, and mixtures thereof, is administered to reduce, prevent or treat a mammal at high risk far or suffering from a cardiovascular disease.

A further object of the present invention is to provide compositions and methods for the i eduction, prev ention, and/or ti eatment of caidiovascular diseases or disorders wherein an effective amount of a composition having at least one a tσcσtricnol, flavonoid, and mixtures thereof, is administered to reduce, prevent, or treat a mammal at high risk for or suffering from a cardiovascular disease,

A still further object of the present invention is to provide compositions and methods for the reduction, prevention, and/or treatment of cardiovascular diseases or disorders wherein an effective amount of a composition having at least one limocitrin derivative, quercetin derivative, naturally occurring poiymethoxyaflavone, tocotrienol, and mixtures thereof, is administered to a mammal to lower serum cholesterol, apo-B, and/or LDL cholesterol.

Another object of the present invention is to provide compositions and methods for the reduction, prevention, and/or treatment of cardiovascular diseases or disorders wherein an effective amount of a composition having at least one limocitrin derivative, quercetin derivative, naturally occurring pσh methoxyaflavσπe, tocotrienol, and mixtures thereof, in combination with a cholesterol-lowering drug, is administered to a mammal to lower serum cholesterol, apo-B, and/or LDL cholesterol

Another object of the present invention is to provide compositions and methods for the reduction, prevention, and/or treatment of cardiovascular diseases or disorders wherein an effective amount of a composition having at least one limocitrin derivative, quercetin derivative, naturally occurring poiymethoxyaflavone, tocotrienol, and mixtures thereof, in combination with a pharmaceutical drug including anti-platelets agents, beta-adrenergic blocking agents, nitrates or calcium channel blockers.

Further objects and advantages of the present invention will become apparent from the following description.

4. DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the use of at least one of limocitrin derivative, quercelin derivative, polymethoxyflavσne. tocotrienol and mixtures thereof alone or in combination with at least one cholesterol-lowering drug for the treatment of cardiovascular diseases or disorders. Limociii in occurs in the peel of lemon as limocitι iπ-3-Oglucσside, and can be produced from the 3-glycoside by enzymatic and acid hydrolysis (Horowitz et al,. J. Org. Chem., Volume 25. 21 885-21 887, 1 960) or by a chemical synthesis procedure such as reported by Dryer et al., Tetrahedron, Volume 20, 2977-2983, 1 64. Two limocitrin analogues, limocitrin 3,7.4'-trimethylether and limσcitrin-3,5,6-4'-tetramethylether, also occur in orange peel (Tatum et al., Phytoche istry, volume II, 2283-228S, 1972). Several polymethoxyflavones were tested and found to be active as inhibitors of apolipoprotein B (apoB) production and had negligible cytotoxicity in the human liver carcinoma cell line HepG2. It has been shown that humans with coronary heart disease (CAD) have higher levels of apoB in their blood. ApoB concentrations also reflect the number of LDL, and VLDL (very low density lipoprotein) particles in arteries. Administering polymethoxylatedflavone of the invention_. to a mammal results in a reduction in the amount of substances in the blood which contribute to CAD. such as for example apoB, LDL, cholesterol, etc; preferably reduction of the serum, plasma, or whole blood concentration or in vivo amounts of these substances. Preferably the concentration or in vivo amount of these substances is reduced to normal lev els typically found in such a mammal. Also, preferably, the polymethoxylatedflavone of the present invention are administered in amounts which produce little qr no cytotoxicity, more preferably where no cytotoxicity is produced.

By way of definition, a polymethoxylatedflavone is a flavone substituted with methoxy groups, preferably at least 2, more prefeiably at least 3 , more preferably at least 4, more preferably 4-8, and most preferably 4-7 methoxy groups and optionally substituted by one or oie hydroxy groups, preferably 1 -3, and more preferably 1 -2 hydroxy groups.

Four compounds of the present invention were synthesized from the lemon flavonoid limocitrin (3',S-dimethoxy-3,5,7,4'-tetrahydroxyfIavone) for use in the present invention: limocitrin-3 ,7,4'-trimethylether (5-hydroxy-3,7,8,3',4'-pentamethoxyflavone); limocitrin- 3,5.7,4'-telramethylether (3 ,5,7,8,3'4'-hexamethoxyflavone); and limocitrin-3,7,4'- trimethylether- -acetate.

A number of methoxylated flavones, most of which occur naturally in citrus, have been found to be useful in the pteseut invention. Also included are substituted derivatives of quercetin. Tile compounds in these gioups include 5-des elhylnobilelin (5-hydιoxy-

6, 7, 8, ',4 '-pentamethoxy lavone); tetra-O-methylisosculcllarcin (5,7,8,4'- tetrame lli oxy Flavone), 3, 5, 6 ,7, S, 3' , 4' -he pta methoxy flavone, nobiletin (5, 6, 7, 8,3', '- hexa ethoxyriavone), langerelin (5,6,7,8,4'-ρenlamethoxyflavone); sinensetin (5, 6,7, 3', '- pentamethoxy fla one), 5 -desmelhy I sinensetin (5-hydroxy-6,7,3',4'-lelramelhoxyfl vo'nc); quercetin lclramelhylelher (5-hydroxy-3,7,3',4'-tεltamethoxyflavone); quercetin 3,5- dimclhylelher-7,3',4'-tribeπzylether, quercetin pcnlamethyl ether (3, , 7,3', '- pen lame thoxy la one), quercel(n-5,7,3',4'-tetramethylether-3 -ace tale; quercelin- 5, 7 ,3', '- lelramethylether (3-hydroxy-5,7,3',4'-letramethoxyflavone).

The basic structutes for limicitriπ derivatives and 5-desmethylsinensetin are represented belovv

H Jcrtrin-3,7,4'-trin-eth\,leLher limocitrin- 3, 7, 4' -uimelhylelher- 5- acetate

H

limociti in- , .7.4' - teti amelh, lether 5-desmelhylsinenselin Examples of tocotrienol compounds useful in the present invention include, but are not limited to. are alpha-tocotrienσl, gamma-tocotrienol, delta-tocotrienol, and mixtures thereof.

Examples of cholesterol-lowering drugs for the treatment of cardiovascular diseases or disoiders useful in the present invention include, but are not limited to, are cholestyramine, cσlestipol. clσfibrate. gemflbrσzil or lovastatiπ,

The methods of the present invention may be administered to any mammal, Most preferably, the polymethoxylatedflavone useful in the methods of the present invention are administered to humans.

In another aspect of the present invention, the polymethoxylatedflavone may be formulated into a pharmaceutical preparation by a conventional method usually employed in the art,

Dosages for the compositions of the present invention may be formulated into pharmaceutical preparations for administration to mammals for reduction, prevention, and treatment of cardiovascular diseases. Examples, not limited thereto, of cardiovascular disease treatable b> the compositions of the present invention include hypercholesteroleiπia, hyperlipidemia. atherosclerosis, thrombosis, m ocardial infarction, etc

Man)' of the limocitrin derivatives, quercetin derivatives, naturally-occurring polymethoxv fla σnes, tocotrienol compounds and mixtures thereof may be provided as compounds w ith pharmaceutically compatible counterions , a form in which they may be soluble, Countcrions for the purposes of this inN'cntion include, for example, hydrophilic and hydrophσbic agents.

The polv methoxylatedflavone can be administered by a variety of routes, including oral, transdermal. rectal, intrarticular, intravenous, and intramuscular introduction. However, it should be understood that the amount of the polymethoxylatedflavone actually administered ought to be determined in light of various relavent factors including the condition to be treated, the chosen route of administration, the age and weight of the individual paiieni. and the seventy of the patient^'s condition, and therefore, the doses given herein should not be construed to limit the scope of the invention in any way. The polymethoxylatedflavone useful in the present invention may be administered in a pharmaceutically or physiologically acceptable carrier. The pharmaceutically or physiologically acceptable carrier is any solvent with which the polymethoxylatedflavone is compatible and which is non-toxic to individuals treated at the amounts administered, A variety of delivery systems for pharmacological compositions may be employed including, but not limited to, liposomes and emulsions. The pharmaceutical compositions also may comprise suitable solid or gel phase carriers or excipients. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

Formulations suitable for oral administration include liquid solutions of the active compound or compounds dissolved in a diluent such as, for example, saline, water, PEG 400; solid preparations such as capsules or tablets, each containing a predetermined amount of the active agent as solids, granules, gelatins, suspensions, and/or emulsions,

Formulations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile solutions which contain buffers, antioxidants. and preservatives. The formulations may be in unit dose or multi-dose containers.

Dosages administered are any effective amount of a polymethoxylatedflavone which will, when given for the treatment, prophylactically or therapeutically. reduce or prevent cardiovascular diseases by reducing levels of substances which contribute to cardiovascular diseases to normal or near normal levels in the blood or in vivo. By way of definition substances which contribute to . cardiovascular diseases, include but are not limited to apσprotein B, low density lipoproteins, very low density lipoproteins, cholesterol, etc.

Fatient dosages for oral administration of flavonoids range from about 1 - 1000 mg/day, commonly 1 -500 mg/day, and typically 1 - 100 mg/day. Stated in terms of patient with a 70 kg body weight, usual dosages range from about 0.01-15 mg/kg/day, commonly from about 0.01 -7.0 mg/kg/day, and typically from about 0.01 -2.0 mg/kg/day,

Patient dosages for oral administration of synthetic flavonoid analogues range from about 2000-5000 mg/day. commonly from about 1 000-2000 mg/day. and typically from about 500-1500 mg/day.

Patient dosages for oral administration of limocitrin derivatives, quercetin derivatives, naturally-occurring polymethoxyflavones. and tocotrienols range from about 1 - 1000 mg/day, commonly about 1 -500 mg/day, and typically from about 1 - 100 mg/day ,

Patient dosages for oral administration of synthetic limocitrin derivatives range from about 200-500 mg/day, commonly about 1000-2000 mg/day, and typically from about 500- 1500 mg/day.

Patient dosages for oral administration of naturally-occurring polymethoxyflavones range from about 1- 1000 mg/day, commonly from about 1 -500 mg/day, and typically from about 1 - 100 mg/day. Stated in terms of patient body weight, for about 70 kg body weight, usual dosages range from about 0,01 - 15 mg/kg/day, commonly from about 0.01 -7.0 mg/kg/day, and typically from about 0.01 -2,0 mg kg/day.

Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety which are sufficient to maintain the anti-prohferative and antioxidative effects of the disease being treated.

For local administration, the composition can be administered by injection directly into a tissue, often in a depot or sustained release formulation.

The follow ing examples illustrate the use of the invention for lowering substances which contribute to cardiovascular diseases and disorders. They are intended to further tllusti ate the invention and are not intended to limit the scope of the invention as defined by the claims.

EXAMPLE 1 Limocitrin derivatives, quercetin derivatives, naturally occurring polymethoxyflavones, and tocotrienols were tested for cholesterol lowering activity in HepG2 cells. Confluent HepG2 cells were preincubatcd for 24 hours in a lipoprotein-free medium (Minimum Essential Medium (MEM), Life Technologies, Burlington, Canada) in which the fetal bovine serum (Life Technologies, Burlington, Canada) was replaced by bovine sei um albumin (BSA, Life Technologies, Bui li gton. Canada ) to inhibit cell proliferation and to stimulate synthesis of cholesterol-containing lipoproteins. Cells were subsequently incubated for another 24 hours in the same medium in the presence or absence of limocitrin derivatives, quercetin derivatives, naturally occurring polv methoxv flavones, or tocotrienols (See Table 1) al the highest concentrations sustaining about 100% ± 10% cell viability, as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphen.vltetrazolium bromide (MTT) viability assay. The lest compounds were added to the cell culture medium after solubilization in dimethyl sulfoxide (DMSO) and filter sterilized. The final concentration of DMSO in culture media did not exceed 0.3% by volume, to prevent anv effects of DMSO on apo B metabolism. For i each experiment, equal concentrations' of solvent were added to all treatment media and to a control medium (without tested compounds). At the end of the incubation, culture media were collected and the concentration of the LDL structural protein, apo-B, was measured by ELISA. The competitive Elisa assay kit for determination of human apo B (Ortho Diagostics, LaJoIIa, CA) measures medium apo B concentrations w ithi-n the range 0.4-0,0 1 μg/mL. In this assay, microtitei 96-well plates ai c coated w ith monoclonal antibody against human apo B, which binds apo B epitope from human LDL (standard curve, control sera with high and low content of human apo B) or from LDL-like lipopiotetn-s secreted by HepG2 cells (tested media, usually 2 x diluted with Ca²⁺- and Mg^2" -free phosphate buffered saline (PBS)). After the exposure, a fraction of antibody is blocked and the remaining amount is captured by a subsequent incubation with conjugate containing a secondary antibody (against anti-human apo B) coupled with horseradish peioxidase. The follow ing incubation with horseradish pei oxidasc substrate, o-pheπyleπcdiamine, produces a \ ellow color change proportional to the amount of apo B present, and the optical density of samples is measured at 490-492 nm against the blank PBS/conjugate wells. To detei nine hether evaluated compounds cross- tcact with Elisa assay, in separate stud)', various doses of compounds were tested in presence of cell-fiee culture media.

Amount of lipoprotein -associated apo-B in the media met apo-B secretion) w as detei mined as described above, and calculated per mg cell protein. Net apo B secretion was also expressed as percent of control (amount of apo B in medium from cells incubated without tested compounds).

Table 1 : IC,„ concentrations were 2.5 μg/mL for tangeretin, 4.9 μg/mL for nobiletin, 7.8 μg/mL for heptamethoxyflavoπe and 17.8 μg/mL for sinensetin.

Claims

The three naturally occurring polymethoxyflavones tested have a dose-response inhibitory effect on apo-B production. These results along with other preliminary studies indicate that the following compounds when used either alone or in combination have inhibitory effects on apo-B production: the limocitrin and quercetin derivatives: limocitrin- 3,7,4'-trimethylcther (5-hydroxy-3,7,8.3',4'-pentamethoxyflavonε); limocitriπ-3, 5,7,4'- tetramethylether (3 ,5,7,8,3',4'-hexamethoxyflavone); limocitrin-3,5.7,4'-tetraethy!ether (8,3'- dimethoxy-3,5,7,4'-hexamethoxyflavone): limocitrin-3,7,4'-trimethylether-5-acetate; quercetin tetramethy [ether (5-hydroxy-3 ,7,3',4'-tetramethoxyflavone); quercetin-3 ,5- dimethylether-7,3',4'-tribenzyl ether; quercetin pentamethyl ether (3, , 7,3', 4'- peπtamethoxy flavone); quercetin-5,7,3',4'-tetramethylether-3-acetate; and quercetin-5,7,3',4'- letramethylether (3-hydroxy-5 ,7,3',4'-tetramethoxyflavone); and the naturally occurring polymethoxyflavones. 3,5,6,7,8,3',4'-heptamethoxyflavone; nobiletin (5, 6, 7, 8.3'.4'- hexamethoxyflavone); tangeretin (5,6.7,S,4'-peπtamcthoxyflavone); 5-desmethylnobiletin (5- bydιoxy-6,7,8,3',4'-peπtamethσx" flavone); tetra-O-methylisoscutellarein (5,7,8,4'- tetramethoxy flavone); 5-desmethylsinensetin (5 -hydrσxy-6, 7, ',4'- tetramcthoxy flavone); and sinensetin (5.6.7.3'.4'-pentamethoxyflavone).

The present invention is not to be limited in scope by embodiments disclosed in the examples that are intended illustration purposed and any methods which are functionally equivalent are within the scope of the invention. Various modifications of the invention in addition to those shown and described herein will become apparent to one of ordinary skill in the from the foregoing description, Such modifications are intended to fall within the scope of the appended claims. The various publications cited herein are herein incorporated by reference in their entirety. We claim:

I . A method for reducing the level of substances in a mammal which contribute to caidiovascular diseases or disoi deis compnsing administering to a mammal in need of such treatment, an effective amount of a pσlymethσxyflavone to reduce the production of substances in a mammal which contribute to cardiovascular diseases and/or disorders.

2. The method of claim I wherein the substances are selected from the group consisting of apolipoprσtein B, cholesterol, low density lipoprotein, very low density lipoprotein and mixtures thereof.

3. The method of claim 2 wherein the substance is apolipoprotein.B,

4. The method of claim 1 wherein said mammal is a human.

5. The method of claim 1 wherein in said pσlymethσxyflavone is selected from the group consisting of limocitrin-3 ,7,4'-trimethylether (5-hydroxy-3,7,8,3',4'-pentamethoxyflavone), limocitriπ-3 ,5,7,4'-tetramethylether (3 ,5.7,8.3',4'-hexamethoxyflavoπe), limoci trin-3.5,7,4'- tetraeth [ether (8.3-dimethoxy-3,5,7,4'-tetraethoxyflavone), limocitrin-3,7,4'-trimethylether- 5-acetate, and mixtures theieof.

6. The method of claim 1 wherein said polymethoxyflavone is selected from the group consisting of 5-desmethyInobiletin (5-hydroxy-6,7,8,3',4'-pentamethoxyflavone); tetra-O- meth)'lisσscutellarein (5,7,S,4'-tetramethoxyflavone), sinensetin (5,6,7,3/4'- pentamethoxyflavone), 5-desmethylsinensetin (5-hydroxy-6,7,3',4'-tetramethoxyflavone), quercetin tetramelhylether (5-hydiOxy-3.7.3'.4'-tetramethoxyflavoπe), quercetin-3 ,5- dimethylether-7,3'.4'-tribenzy! ether, nobiletin (5,6,7,8,3',4'-hexamethoxyflavone). quercetin pentamethyl ether (3, 5,7,3'. '-pentamethoxyflavσπe), tangeretin (5.6,7,8,4'- pentamethoxyflavone), quercetin-5,7,3',4'-tetramethyl ether-3-acetate, 3, 5, 6,7, 8,3', 4'- heptamethoxyflavone, 5,7,3',4'-tetramethylether (3-hydroxy-5,7,3',4'-tetramethoxyflavone), and mixtures thereof.

7. The method of claims 5 or 6 further comprising administering at least one tocotrienol w herein said tocotrienol is selected from the group consisting of alpha-tocotrienol, gamma- tocotrienol, dεlta-tocotrienol, and mixtures thereof.

8. The method of claims 5 or 6 further comprising administering at least one pharmaceutical drug used in inhibiting cardiov ascular diseases.

9. A pharmaceutical composition suitable for administering to a mammal at risk or suffering from a cardiovascular disease or disorder comprising an effective amount of at least one polv methoxyflavαne to reduce the production of substances in a mammal which contribute to cardiovascular diseases or disorders selected from the group consisting of limocitrin-3,7,4'-trimethylether (5-hydroxy-3,7,8,3',4'-pentarnethoxyflavσne), limocitrin- 3.5.7.4'-tetramethylether (3,5.7.8,3',4'-hexamethoxyflavone), limocitrin-3.5,7,4'- tetraethylether (8.3-dimethoxy-3.5,7,4'-tetraethoxyflavone). limocitrin-3.7,4'- tri eth) lether-5-acetate, and mixtures thereof.

1 0. A pharmaceutical composition suitable for administering to a mammal at risk or suffering from a cardiovascular disease or disorder, said composition comprising an effective amount of at least one polymethoxyflavone to reduce the production of substances in a mammal which contribute to cardiov ascular diseases or disorders selected from the group consisting of 5-desmcthv Inobiletin (5-hydroxy-6,7,8,3',4'-pentamethoxyflavoπe); tetra-C7- methyhsoscutellarein (5.7,8.4''tetramethoxyflavone), sinensetin (5,6,7,3/4'- pentamethoxy flavone), 5-desmeth I sinensetin (5-hydroxy-6,7.3',4'-tetramethoxyflavone), quercetin tetramethylether (5-hydroxy-3,7,3',4'-tetramethoxyflavone). quercetin-3,5- dimethv lether-7,3',4'-tribenzyl ether, nobiletin (5,6,7,8,3',4'-hexamethoxyflavone), quercetin pentamethyl ether (3,5,7.3',4'-pentamethoxyflavone), taπgeretiπ (5,6,7,8,4'- pentamethoxy flavone), quercetiπ-5.7,3',4'-tetramethyI ether-3 -acetate, 3, 5, 6, 7.8,3', 4'- heptamethoxyflavone. quercetin-5.7,3',4'-tetramethyI ether (3-hydroxy-5.7,3'.4'- tetiamethoxv fiav one), and mixtures theieof.

1 1 . The composition of claims 9 or 10 further comprising at least one tocotrienol wherein said tocotrienol is selected from the group consisting of alpha-tocotrienol, gamma-tσcotrienol, delta-tocotrienσl. and mixtures thereof.

12. The composition of claims 9 or 10 further comprising administering at least one pharmaceutical drug used in inhibiting cardiovascular diseases.