Abstract:
The invention provides a method for increasing serum high density lipoprotein (HDL) cholesterol levels, and lowering serum levels of Lipoprotein (a) and C-reactive protein, as well as a suitable composition comprising a) a vegetable protein, b) a natural antioxidant compound, c) a natural anti-inflammatory compound, d) a bioavailable source of selenium, e) a natural blend of long chain aliphatic alcohols termed policosanol; and f) a natural bioavailability enhancer.

Description:
CROSS REFERENCE TO RELATED APPLICATIONS  
       [0001]     This application claims the benefit of U.S. Provisional patent application No. 60/594,042 filed on Mar. 7, 2005. 
     
    
     BACKGROUND INFORMATION  
       [0002]     Compositions containing natural compounds and nutrients that efficaciously and safely enhance high density lipoprotein cholesterol (HDL-C) levels, are described. These compositions also favorably influence the markers of inflammation and support cardiovascular health and wellness in humans and animals.  
         [0003]     Cardiovascular diseases (CVD) continue to remain the leading cause of morbidity and mortality in the developed world, with coronary heart disease (CHD) being rated as the number one killer, and stroke following as the leading third, in the United States. Over the years, research has established the link between dietary fats (lipids), lipid transport and metabolism in the body, atherosclerosis (the progressive narrowing of the arteries over time), and cardiovascular disease. Lipids present in the blood and tissues of the body include cholesterol, cholesterol esters, triglycerides, and phospholipids.  
         [0004]     High levels of low-density lipoprotein cholesterol (LDL-C) are particularly correlated with atherosclerosis and cardiovascular disease, based on evidence from clinical studies and epidemiological analysis. Levels of inflammatory markers such as C-reactive protein, homocysteine and low-density lipoprotein (LDL) cholesterol levels are elevated in persons at risk for cardiovascular events.  
         [0005]     Currently, the statins are the first choice drugs when treating dyslipidemias, especially in patients with hypercholesterolemia alone or accompanied by hypertriglyceridemia.  
         [0006]     Most therapeutic approaches target reducing LDL. However, studies in animal models show that over-expression of apolipoprotein (apo) A-1, the major HDL lipoprotein, inhibits progression and induces regression of atherosclerosis. Therefore HDL is an important target for therapeutic intervention, and low HDL-C levels are a modifiable risk factor for cardiovascular disease.  
         [0007]     Current medical opinion considers inflammation of atherosclerotic plaques, and the subsequent formation of blood clots on the surface of these plaques, as critical events that lead to most atherosclerosis induced cardiovascular events.  
         [0008]     As National Cholesterol Education Program (from the National Institutes of Health) guidelines have been changed to include a global measure for CHD risk, risk status and treatment measures adopted for some patients, may be different as compared to the earlier guidelines. The number of patients with cholesterol levels that can be classified as abnormal, has now tripled. These guidelines recommend complete lipoprotein profile (total, LDL, HDL, triglycerides) as preferred screening for assessing CHD risk status. LDL remains a primary target of cholesterol-lowering therapy, along with increased emphasis on optimal HDL levels.  
         [0009]     Statins are the most powerful drugs for lowering LDL, facilitating dose-related reductions in LDL ranging from 20-60%. Fluvastatin is reported to be the least potent, decreasing LDL levels by only 22-36% at the maximum recommended dosage. Rosuvastatin is reported to be the most potent, reducing LDL levels by up to 65% (in a dose range of 20-80 mg/day), in clinical studies. Statins also have moderate effects on HDL, raising levels by approximately 5%, and decrease triglyceride concentrations to a maximum of about 30%.  
         [0010]     Unfortunately, the mechanism of action of statins through inhibition of the mevalonate pathway inhibits the biosynthesis of vital biochemical products of this loop As a result, myalgia and myopathy occur in 2% and 0.5% of patients, respectively, with less than 0.1% of cases progressing into rhabdomyolysis, which may be associated with acute renal failure.  
         [0011]     Besides statins, other current lipid-altering agents that lower LDL-C primarily through increased hepatic LDL receptor activity include, bile acid sequestrants/resins and cholesterol absorption inhibitors such as ezetimibe. These are also associated with side effects.  
         [0012]     In recent years there has been significant research on the use of safer efficacious natural alternatives and nutritional materials to address lowering LDL-C, raising HDL-C and to help individuals with various types of dyslipidemias. A few examples from published literature are cited here.  
         [0013]     Plant sterols/stanols (Metabolism. 52(11):1478-83) polyphenols, natural antioxidant herbal extracts such as curcuminoids from turmeric (Indian J Physiol Pharmacol.; 36(4):273-5), viscous fiber such as oat bran (Ann Nutr Metab. 47(6):306-11), saponin-rich seed extracts from fenugreek (J Assoc Physicians India; 49:1057-61), and seed proteins such as soy protein (Arteriosclerosis, Thrombosis, and Vascular Biology. 2003; 23:2241) have also been shown to be beneficial in lipid lowering.  
         [0014]     Guggulsterones the biologically active components of the resin of guggul ( Commiphora mukul ) used in traditional Ayurvedic medicine to treat inflammation, arthritis, cardiovascular conditions and obesity were recently shown to be antagonist ligands for the bile acid receptor FXR, which is an important regulator of cholesterol homeostasis in the body. (Science; 296:1703-6).  
         [0015]     Policosanol (Am HeartJ; 143(2): 356-65) is a natural mixture of higher aliphatic alcohols, found in plant waxes. Sugarcane wax is the common source. The components of policosanol include 1-octacosanol, 1-dotriacontanol, 1-tetracosanol, 1-tetratriacontanol, 1-hexacosanol, 1-heptacosanol and 1-nonacosanol. This mixture of alcohols is clinically proven to be effective in maintaining normal cholesterol levels.  
         [0016]     Coconut protein is shown to increase HDL (Indian J Exp Biol. 2001 October; 39(10):1028-34).  
         [0017]     L-arginine present in coconut protein is reported to favorably influence blood lipid levels (Indian J. Exptal Biology Vol. 42, January 2004, pp. 53-57). L-arginine favorably influences serum homocysteine levels as well. Homocysteine is a marker of cardiovascular abnormalities. (Journal of Nutrition, 135, pp 212-217).  
         [0018]     Curcuminoids extracted from Zingiberaceae species are known natural antioxidants and favorably influence cholesterol levels (Indian J Physiol Pharmacol. 1992 Oct; 36(4):273-5).  
         [0019]     Boswellic acids from Boswellia species are reported to lower cholesterol levels in animal models. (Indian J. Pharmacol.—18: 182-183, 1986)  
         [0020]     Selenium-enriched garlic is reported to increase HDL-C levels, (Wei Sheng Yan Jiu. 2002 April; 31(2):93-6). Dietary intake of non-fish Se had a positive correlation with HDL cholesterol (Trace Elem. Med Biol. 2002; 16(2):83-90)  
         [0021]     Thus natural compounds and nutritional materials that have beneficial effects on blood lipid levels, enhance HDL-C levels and support cardiovascular health are available for use as alternatives or to complement drug therapy.  
       SUMMARY OF THE INVENTION  
       [0022]     Compositions containing natural compounds and nutrients that efficaciously and safely enhance HDL cholesterol levels, are described. These compositions also favorably influence the markers of inflammation and support cardiovascular health and wellness in humans and animals. 
     
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0023]      FIG. 1  shows the effect of Formulations 1 and 2 on Total Cholesterol in an animal model.  
         [0024]      FIG. 2  shows the effect of Formulations 1 and 2 on triglycerides level in an animal model.  
         [0025]      FIG. 3  shows the effect of Formulations 1 and 2 on LDL level in an animal model.  
         [0026]      FIG. 4  shows the effect of Formulations 1 and 2 on HDL level in an animal model.  
         [0027]      FIG. 5  shows the effect of Formulations 1 and 2 on VLDL level in an animal model. 
     
    
     DETAILED DESCRIPTION  
       [0028]     Synergistic Compositions prepared with natural compounds and nutritional ingredients are described in the present invention. These compositions favorably influence HDL-C, Lipoprotein (a), and LDL-C levels, have beneficial effects on inflammation, and provide antioxidant action.  
       EXAMPLE 1  
       [0029]     Nutritional supplement composition to enhance HDL-C levels The ingredients as listed below were formulated in pharmaceutically acceptable vehicle.  
                                             Formulation 1                                  Commiphora mukul  Extract(standardized to   34   g (68.5% w/w)       contain 2.5% guggulsterones)         Policosanol     600   mg (1.2% w/w)       Garcinol (40% extract from  Garcinia  species)   15   g (30.2% w/w)                 Animal dose administered: 250 mg/kg body weight             
 
         [0030]    
       
         
               
             
               
               
               
             
           
               
                   
               
               
                   
               
               
                 Formulation 2 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Coconut protein: 
                 33.3 
                 g (76.8% w/w) 
               
               
                 Curcuminoids (from Turmeric 
                 5.0 
                 g (11.53% w/w) 
               
               
                 ( Curcuma longa  ) roots) 
               
               
                 Acetyl-11-keto-beta-boswellic acid 
                 5.0 
                 g (11.53% w/w) 
               
               
                 Gamma-glutamyl-methyl-L- 
                 50 
                 mcg Selenium equivalent 
               
               
                 selenocysteine 
               
               
                   
               
               
                   Animal dose administered: 90 mg/kg body weight    
               
             
          
         
       
     
       EXAMPLE 2  
     Efficacy and Safety Study in Animal Model  
       [0031]     The compositions listed in paragraph 21 was tested in an animal model study Study design: Healthy female mice were divided into three groups of 40 mice each (Group A: Control group, Group B: Administered test formulation 1, Group C: Administered Test Formulation 2). Each group was divided into four sub-groups of ten mice each. The segregation was random.  
         [0000]     Group B received formulation 1 suspended in 0.5% Carboxymethyl cellulose (CMC) 250 mg/kg body weight along with normal diet.  
         [0000]     Group C received Test formulation 2 suspended in 0.5% CMC-90 mg/kg body weight along with normal diet.  
         [0032]     The groups were further divided into three sub-groups each. One sub-group from each group was randomly selected to determine blood lipid profile (Total cholesterol, HDL cholesterol and triglycerides). The remaining sub-groups from each group were analyzed at 4 week intervals for blood lipid profiles (4 weeks, 8 weeks, 12 weeks).  
         [0033]     Results:  
         [0034]     Throughout the experimental period of twelve weeks all the animals tolerated the treatment well. They did not show any change in behavior in feeding, drinking or grooming. They remained alert and were responsive to external stimuli. All the animals survived during the course of treatment.  
         [0035]     Statistical analysis of the data presented in  FIGS. 1-4  revealed the following:  
         [0000]     a) Both Formulation 1 and Formulation 2 are effective in favorably influencing blood lipid levels in experimental animals.  
         [0000]     b) Periodical analysis through 4, 8 and 12 weeks using inter-group controls indicates that during the first 4 weeks of treatment, formulation 1 has better efficacy in lowering triglycerides, LDL-C, total cholesterol and in increasing HDL-C.  
         [0000]     c) At the end of the eighth week, the trend continued as above.  
         [0000]     d) At the end of 12 weeks however, formulation 2 was found to more efficaciously reduce total cholesterol and LDL-C as compared to formulation 1. The values of the other parameters were more or less identical for both formulations.  
         [0036]     Based on these results, the long term efficacy in lowering total cholesterol and LDL-C of formulation 2 is superior to that of formulation 1. Also formulation 2 efficaciously lowers total cholesterol and LDL-C and raises HDL-C at a lower dose as compared to formulation 1. The dose of formulation 2 needed is 2.8 times lower than that of formulation 1.  
       EXAMPLE 3  
       [0037]     Evaluation of The safety and efficacy of synergistic compositions of natural compounds and nutritional ingredients in supporting healthy blood lipid levels and cardiovascular health and wellness in human subjects:  
         [0038]     A formulation was prepared by dispersing potentially effective amounts of coconut protein, plant based antioxidant and anti-inflammatory compounds, a bioavailable form of nutritional selenium, policosanol, and a natural bioavailability enhancer, in a pharmaceutically acceptable carrier. A suitable oral dosage form was prepared.  
                                                                     Active Ingredient   Amount/dose                                        Coconut protein   400   mg           Curcuminoids (from  Curcuma longa  roots)   50   mg           Acetyl-11-keto-beta-boswellic acid   50   mg           Gamma-glutamylmethylselenocysteine   200   mcg             Policosanol     5   mg           Black pepper Extract (Bioperine (RTM of   5   mg           Sabinsa Corporation New Jersey, USA)           standardized to contain 95% to 98% piperine)                      
 
         [0039]     9 individuals (8 males and 1 female) 25 to 55 years of age with total cholesterol levels at baseline ranging from 168 mg/dL to 251 mg/dL participated in the study. The study examined total cholesterol, HDL cholesterol, triglycerides, LDL cholesterol, VLDL cholesterol. Lipoprotein (a), malondialdehyde (a marker of oxidative stress), and C-reactive protein levels (high sensitivity C-reactive protein) in these subjects before treatment, and after 34, 68 days of treatment with 2 tablets/day of the composition  
         [0040]     Blood samples were taken at Day 0, Day 34, and Day 68 and evaluated for blood lipid profile and inflammatory markers.  
         [0041]     Total cholesterol levels decreased from baseline levels after treatment in all volunteers. The percentage reduction ranged from 10% to 40%. HDL-C levels increased in 7 out of 9 volunteers. The percentage increase from baseline was 1% to 10%. The levels of triglycerides decreased after treatment by 7% to 45% from baseline levels, LDL-C levels decreased from 10% to 45% and VLDL levels decreased by 7% to 45%.  
         [0042]     Mean levels of the inflammatory marker C-reactive protein (hsCRP) and malondialdehyde were reduced from baseline levels after 68 days of treatment. The reduction in hsCRP was from an initial value of 1.0±0.5 mg/dL to 0.7±0.3 mg/dL. Malondialdehyde levels decreased from an initial value of 1.2±0.5 mg/dL to 0.7±0.3 mg/dL. Lipoprotein (a). (Lp (a)) levels were reduced by a mean of 5% from the pre-treatment levels.