Lycopersicum esculentum plant extracts and uses thereof

FIELD OF INVENTION

The present invention pertains to nutritional or pharmaceutical compositions comprising extracts or concentrates of plants and the mixtures thereof belonging to Lycopersicum with specific reference to Lycopersicum esculentum. The present invention further relates to extracts which are isolated from different parts of Lycopersicum esculentum plant, the preparation of such extracts and the medicaments containing said extracts The invention further relates to screening and characterization of extracts for their activity in preventing, mitigating, or treating cardiovascular disorders to achieve the lowest possible cardiac risk. Furthermore, the invention relates to the use of the extracts as a supplement or a medicament useful in the treatment/alleviation or prevention of cardiovascular diseases.

BACKGROUND OF INVENTION

Cardiovascular disease is a group of diseases affecting heart and blood vessels (arteries). Increased incidence of cardiovascular disease in the recent decades can be attributed to the lifestyle and diet as the disease has dietary basis (Bogin, 1997; Hamilton et al., 1988).

The cardiovascular disease is one of the main causes of mortality in human beings, which needs major breakthrough discoveries from natural products that can be helpful in preventing risks of cardiovascular disease.

CLASSIFICATION OF CARDIOVASCULAR DISEASE

Cardiovascular disease is broadly categorized into - Coronary heart disease, high blood pressure, heart stroke, heart failure and vascular inflammation. Coronary heart disease refers to the lack of blood supply to the myocardium (heart muscles) due to the accumulation of cell debris containing cholesterol and fatty acids, fibrous connective tissue and Calcium, which are collectively called atheromatous plaques. High blood pressure (hypertension) is a medical condition wherein the blood pressure reading rises to 140/90 mmHg or above. Prolonged hypertension increases the risks of heart attacks and heart failure. Stroke results in the poor supply of blood to the brain and are due to either blockage or rupturing of the blood vessels that connects the brain. Stroke is further classified into four types, cerebral thrombosis and cerebral embolism (blockage of blood vessels) and cerebral and subarachnoid haemorrhages (rupture of blood vessels). Normal blood pressure with respect to cardiovascular risk should be close to 120/80 mm Hg.

THE ETIOLOGY OF CARDIOVASCULAR DISEASE

Over the past three decades, great progress has been made in identifying and correcting risk factors for cardiovascular disease (CVD), such as smoking, high blood pressure and an elevated plasma level of total cholesterol and low-density lipoprotein (LDL) cholesterol. This has led to a significant reduction in the incidence of CVD in several countries. Despite this encouraging result, however, CVD is still the leading cause of death in many nations. A further decline in cardiovascular morbidity and mortality could be achieved by employing a wide, multiple risk factor approach. Many studies opined that risk factors for cardiovascular disease largely consist of dietary basis and also on lifestyle. Despite the information available on risk factors, studies are being undertaken to understand the complexity of biomolecules involved and preventive mechanisms. Studies showed that increased intake of saturated fats leads to the increase in level of low-density lipoprotein (LDL), which enhances the risk of CVD (Bankson et al., 1993). The increased LDL also leads to the formation of atheromatous plaques, subsequently blocking the blood vessels and also increases calcium accumulation in walls of blood vessels (Hattersley, 1993). Low high-density lipoprotein (HDL) cholesterol levels, often associated with elevated plasma triglycerides, play a significant role in the risk of developing CVD (Regnstrom, et al., 1990). This is observed to be one of the most common forms of CVD (Steinberg et al., 1989). Reduced thyroid function increases the incidence of CVD owing to its higher adrenalin production that degenerate the blood vessels.

Hypertension is classified by etiology as being either primary (essential, idiopathic) or secondary. Primary hypertension accounts for 90-95% of cases of hypertension, while the remaining 5-10% of cases are the result of secondary hypertension. Evidence suggests varying combinations of many potential interacting factors causes the disease. Patients with primary hypertension do not appear to share any one, or a specific combination of, suspected etiologic factors. Some of the potential etiologic factors for primary hypertension include:

• Sympathic nervous system dysfunction and/or hyperactivity.

• Renin-angiotensin system defects.

• Sodium transport defects.

• Intracellular sodium and calcium defects.

Other factors have been implicated as either predisposing or contributing to the development of primary hypertension. These include obesity, excessive alcohol use, cigarette smoking, excessive salt intake, stress, and physical inactivity. No clearly established genetic pattern has been established for primary hypertension. However, blood pressure levels appear to have strong familial tendencies.

Etiologies of secondary hypertension include: renal vascular disease (atherosclerotic, thrombotic, embolic stenosis or obstruction, fibromuscular hyperplasia, etc.).

Hypertension is the most important etiologic factor for cardiovascular disease. In adults, systolic blood pressure elevations are usually considered to be more a determinant of cardiovascular risk than are diastolic blood pressure elevations. Hypertension accelerates the development and progression of atherosclerosis (leading to peripheral and coronary vascular insufficiency), and subsequently increases the patient's risk for myocardial infarction. Hypertension also causes left ventricular hypertrophy, which may result in congestive heart failure, ventricular arrhythmias, myocardial ischemia, and sudden death. Hypertension is a major etiology for both dissecting and atherosclerotic aortic aneurysms, and also acts as an exacerbating factor in the progression of these conditions. Retinal vascular narrowing, hemorrhages, exudates, and papilledema are also consequences of hypertension.

Inflammatory vascular diseases are initiated and perpetuated by the interaction of immune cells with cells of the affected vessel wall. This is directed by a network of chemical messengers, which, in a state of vascular health, exist as balanced but opposing forces. The detection of vascular inflammation and monitoring of this activity have long been attempted in systemic vasculitis, and, more recently, in atherosclerosis. Markers of vascular inflammation used thus far have been of limited value; few provide both adequate sensitivity and specificity for any particular disease

Inflammatory markers are predictors of recurrent CVD and death in different settings, including the short-term risk, long-term risk, and risk after revascularization procedures such as percutaneous coronary intervention (PCI), including the risk of restenosis. Several Inflammatory markers which seem to have predictive abilities for Cardiovascular Risk are high-sensitivity C-reactive protein [hs-CRP], serum amyloid A [SAA], white blood cell [WBC] count and fibrinogen. Preferably, only the acute-phase reactants (fibrinogen and CRP) and WBC count have widely available assays. hs-CRP consistently predicts recurrent myocardial infarction independent of troponins, which suggests it is not merely a marker for the extent of myocardial damage. It also may be useful in the estimation of prognosis in patients who need secondary preventive care, such as those with stable coronary disease or acute coronary syndromes (risk of restenosis) and those who have undergone PCI. Elevated hs-CRP levels also seem to predict prognosis and recurrent events in patients with stroke and peripheral arterial disease (Pearson,et.al. 2007).

TREATMENT

Lifestyle modifications with diet therapy include reduced intake of saturated fats, trans-fatty acids, and plant stanol/sterols and viscous fiber and a good exercise program helps to prevent or delay further damage to the cardiovascular system.

Statin drugs, Aspirin and Anticholesterol drugs, such as Mevacor, Cholestyramine, and Clofibrate are prescribed to control the risks of high cholesterol and reduce the chances of strokes and heart attacks (Lita Lee, 2005). Statin drugs and Anticholesterol drugs do lower cholesterol but does not eliminate the risk of heart attacks. Omega-3-fatty acids, carotenoids, vitamins, minerals and some of the herbal drugs are suggested for preventing cardiovascular risks.

The inclusion of botanicals in a nutritional approach presents an inexpensive means of achieving the goal of cost effective management of the said disease condition. However, the effects of the nutritional strategies recommended today are rather modest. Thus, research into novel nutritional strategies preventing cardiovascular risks is needed.

The treatment of cardiovascular disorders is a particularly promising area for botanicals. Most botanicals derive their effectiveness from a mixture of active molecules, acting in concert. Multiple agents attacking multiple targets simultaneously present decided advantages over conventional drugs, which are each based on one compound that produces one action. Plants hold the power to keep the increasing prevalence of cardiovascular disorders, in check, prompting the search and trial of plant extracts to develop a whole new category of natural products. Comprehensive studies on the components of the herb that are responsible for certain indications need to be undertaken to obtain effective medications from this therapy. Also, not necessarily all the ayurvedic extracts are non-toxic.

The use of herbs has shown positive effects on the cardiovascular system. Dry fruits reduce cholesterol and lower the risk of genetic coronary disease due to its rich source in alpha-linolenic acid (ALA). Flavonoid rich fruits and peels have the potential to reduce the risks of cardiovascular disease (Donald R. Yance, 2005).

Lycopersicum esculentum (Tomato) has a high antioxidant capacity due to its high content of vitamin C, vitamin E and lycopene that is a powerful free radical scavenger found naturally. Dietary supplementation of concentrated tomato juice significantly increases lycopene levels and HDL cholesterol (Madrid 2006). It may also reduce cholesterol levels and lipid peroxidation; factors implicated in the initiation of cardiovascular disease (Collins et al 2004) and may decrease the risk of coronary heart disease by improving the serum lipid profile (Ahuja 2006).

Tomatoes contain the steroidal glycoalkaloid tomatine, which has been reported to form strong; insoluble complexes with cholesterol in vitro are effective options for increasing HDL cholesterol levels. Tomatine can reduce dietary cholesterol absorption and plasma levels of cholesterol and triglycerides (Friedman et al 2000).

Other cardioprotective functions provided by the nutrients in tomatoes may include the reduction of low-density lipoprotein (LDL) cholesterol, homocysteine, platelet aggregation, and blood pressure (Willcox et al.2003).

This background information is provided for the purpose of making known information believed by the applicant to be of possible relevance to the present invention. No admission is necessarily intended, nor should be construed, that any of the preceding information constitutes prior art against the present invention.

PRIOR ART:

The related art of interest describes different process for obtaining the extract of Lycopersicum esculentum and thereby describing the utility of the extract in different ways but none of citations disclose the present invention. There is a vital necessity to obtain Lycopersicum extract, which is not only safe but also effectively applicable in curing and preventing cardiovascular disease and its related disorders. The related art will be discussed in order of obvious and noticeable significance to the present invention.

A PCT Application No: WO2008CA530A, entitled "Compositions comprising polyunsaturated fatty acid monoglycerides or derivatives thereof and uses thereof relates to the inventions provided various compounds and compositions comprising polyunsaturated fatty acid monoglycerides and derivatives thereof. These compounds and compositions can be useful as cancer chemopreventive agents. They can also be useful for enhancing solubility of various active agents and enhancing their bioavailability. The claim 69 of the invention relates to the prevention of cardiovascular disease.

But the present invention relates to the function of Lycoperscicum esculentum composite extract and the process in increasing the HDL-cholesterol level by monitoring the regulation of Apo Al marker. Therefore the given citation does not show any similarity with the present invention as the present invention.

A European patent application, bearing the application no. EP2001936384A entitled, "Composition for improving the cell protection comprising a lipophilic antioxidant and hydrophilic antioxidant' relates to the invention relates to a method for improving the cell protection which comprises the administration of a combination of the extracts of the plants Vitis

vinifera and Lycopersicum esculentum with the vitamins C, E, beta-carotene and optionally selenium. Said combination can be used in the prevention of pathological conditions related in part to an overproduction of free radicals like ageing, arteriosclerosis and cancer.

However, the same is nowhere in relation to the application of the instant invention, where it actually works on the application of the Lycopersicum composite plant extract for improvement of cardiovascular health in terms of up regulation of HDL-c.

A US patent application bearing the application no: US2003442908A, entitled "Antithrombotic agents" relates to the therapeutically effective amount of a tomato fruit extract having platelet aggregation inhibiting activity.

But the present invention relates to the function of Lycoperscicum esculentum composite extract and the process in increasing the HDL-cholesterol level by monitoring the regulation of Apo Al marker. Therefore the given citation does not show any similarity with the present invention as the present invention.

A US patent application bearing the application no. US200247583A relates to the invention concerns a composition containing a high level of Vitamin E in encapsulated form, preferably as beadlets or spray dried, and a high level of silicates. The composition is compressible into tablets or caplets that do not leach out Vitamin E into the tablet matrix during compression and/or storage. The composition preferably contains vitamins and minerals in doses optimized for cardiovascular health. Invention relates to a Vitamin formulation comprising as claimed in claim 1, wherein the additional minerals and vitamins are selected from Folic Acid, iron, lycopene etc. The application of the instant invention relates to the effectiveness and activity of a non-toxic, extract to increase the HDL-c levels in the mammalian and human system. Therefore, the citation given does not have any relevance with the application of the instant invention.

The US patent application bearing the application no. US2002187618A entitled, "Cardiovascular promotion and maintenance composition'''' relates to a composition for to assist in the achievement of optimal cardiovascular health. The composition addresses various concerns including low antioxidant status, low levels of essential fatty acids, magnesium, potassium, and elevated levels of homocysteine. It is designated to treat and prevent heart disease and stroke. Invention relates to a cardiovascular maintenance composition comprising an effective amount of 5-100 mg Lycopene. But the present invention relates to the function of Lycoperscicum esculentum composite extract and the process in increasing the HDL-cholesterol level by monitoring the regulation of Apo Al marker. Therefore the given citation does not show any similarity with the present invention as the present invention.

The US patent application bearing the application no. US2001754957A, entitled, "Composition for reducing plasma triglycerides, platelet aggregation, and oxidative capacity" relates composition comprising at least one unsaturated fatty acid, such as an omega-3 fatty acid; pantethine; and an antioxidant selected from the group consisting of Vitamin C, Vitamin E, tocotrienol, at least one carotenoid, at least one flavanoid, coenzyme Q10, and grape seed extract. Such active ingredients may be encapsulated in an encapsulating medium to form micro particles, which may be suspended in an aqueous solution. Such a composition reduces plasma triglyceride levels, platelet hyper aggregation, endothelium dysfunction, and tissue oxidative capacity, and thus reduces the risk of cardiovascular disease. A composition, comprising: (a) at least one unsaturated fatty acid; and at least one of: (b) pantethine; and (c) at least one antioxidant; said at least one antioxidant is a carotenoid - Lycopene.

However, the same is nowhere in relation to the application of the instant invention, where it actually works on the application of the Lycopene plant extract for improvement of cardiovascular health.

The US patent application no. US2002360789A entitled, "Phytonutrient nutritional supplement' relates to the composition and method for correcting a dietary phytonutrient deficiency. The composition includes one or more of the following phytonutrients: lutein, lycopene, epigallocatechin gallate (EGCG), ellagic acid, hesperidin and quercetin. Dietary phytonutrient deficiencies are corrected by administration of these phytonutrients in amounts equal or greater than the amounts of these phytonutrients present in recommended daily servings of fruits and vegetables. As per claim 16 of the invention the method for improving at least one of cardiovascular health and bone health comprising administering a dietary supplement according to claim 1 to a subject in need thereof. The application of the instant invention relates to the effectiveness and activity of a non-toxic, extract to increase the HDL-c levels in the mammalian and human system.

Therefore, the citation given does not have any relevance with the application of the instant invention.

A PCT application no. WO2004IL974A, entitled "A method for fortifying food stuff with phytonutrients and food products obtained thereby" where the invention relates to a health-beneficial amount of tomato phytonutrients. Claim 9. A method according to claim 8 wherein said health benefits are selected from among DNA damage prevention, blood pressure lowering, cardiovascular ailment prevention and free radical scavenging. But the present invention relates to the function of Lycoperscicum esculentum composite extract and the process in increasing the HDL-cholesterol level by monitoring the regulation of Apo Al marker. Therefore the given citation does not show any similarity with the present invention as the present invention.

OBJECT OF THE PRESENT INVENTION

The principle object of the present invention is to provide an active extract and bioactive fraction obtained from different parts of Lycopersicum esculentum plant.

Another object of the invention is to provide a process for isolating bioactive fraction from Lycopersicum esculentum using aqueous, alcoholic and/or hydro-alcoholic and organic solvent, the preparation of such extracts, evaluating bioenhancing/bioavailability of Lycopersicum esculentum extract or bioactive fraction in combination with nutraceuticals or herbal drugs/products to evaluate the Lycopersicum plant extracts, capable of treating cardiovascular disorders in more than one mode of action.

Yet, another object of the invention is to provide composition comprising active principles of Lycopersicum esculentum, and the use of these extracts and constituents for the preparation of nutritional and nutraceutical application.

Still another object of the present invention is to provide Lycopersicum esculentum plant extract, which is easily and safely administrable to children and adults.

SUMMARY OF THE INVENTION

Accordingly, the present invention provides Lycopersicum esculentum plant extract for management of Cardiovascular disease and/ or related disorders in a subject in need thereof, said method comprising step of administering pharmaceutically acceptable amount of standardized Lycopersicum plant composite extract, optionally along with pharmaceutically acceptable additives, to the subject; and a process for enhancing cardiovascular health properties of a Lycopersicum plant extract, said process comprising steps of (a) size-reducing plant parts to obtain powder; (b) extracting the bioactives with a solvent and/or combination of solvents by heating at temperature ranging from 21° to 105° C to obtain a mixture; (c) clarifying the mixture to arrive at clear liquid; (d) concentrating the clear liquid to achieve a concentrated extract; (e) solubilizing the concentrated extract in a solvent and re-concentrating it to obtain further concentrated extract, followed by drying the treated extract to obtain the plant bioactives which have significant role in reducing cardiovascular risk. The invention further provides for uses of the extract to manufacture a medicament for multiple therapeutic uses, as well as other healthful benefits.

BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWING

Figure 1: Metabolite profile of the extract at 254nm of AVDHC27Le (25) 08(100) extract.

Figure 2: Comparative overlay of AVDHC27Le (25) 08(100) extract.

Figure 3: Metabolite fingerprinting of AVDHC27Le (25) 08(100) extract scanned from 200 to 700 nm using Metagrid™ software.

Figure 4: Metabolite profile of the extract at 254nm of AVDHC27Le (65) 08(100) extract.

Figure 5: Comparative overlay of triplicate run of AVDHC27Le (65) 08(100) extract at 254 nm.

Figure 6: Metabolite fingerprinting of AVDHC27Le (65) 08(100) extract scanned from 200 to 700 nm using Metagrid™ software.

Figure 7: Effect of AVDHC27Le (25) 08(100) on the cell viability in HepG2 cells at 24hrs treatment.

Figure 8: Effect of AVDHC27Le (25) 08(100) on ApoAl release in HepG2 at 24 hrs of treatment.

Fig 9: Effect of AVDHC27Le (25) 08(100) on the cell viability in HepG2 cells at 24hrs treatment.

Fig10: Effect of AVDHC27Le (65) 08(100) on ApoAl release in HepG2 at 24 hrs of treatment.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is in relation to efficiency of the bioactive component of the plant extract for therapeutic use, wherein said extract from Lycopersicum esculentum optionally along with healthful or for nutritional and nutraceutical applications.

In one aspect of the invention, there is a provided a prophylactic method for preventing the occurrence of a disease state in a mammal which comprises administering to the said mammal an effective non-toxic amount of an extract from Lycopersicum esculentum as defined herein in the preparation of a comestible (foodstuff) for prophylaxis against the occurrence of cardiovascular disease. Preferably the mammal is human and the said extract comprises a single extract from a plant part of Lycopersicum esculentum or a combination of extracts there from as detailed herein. Thus the present invention further relates to extracts, which may be isolated from fruits of the Lycopersicum esculentum plant, the preparation of such extracts, medicaments comprising such extracts, and the use of these extracts and constituents for the preparation of a medicament.

Some of the embodiments of the present invention will include pharmacognostic evaluation of the botanicals as per the conventional pharmacopoeial standards for ascertaining the quality, purity and efficacy of the herbs. These tests will comprise determination of physico-chemical standards like total ash, water and alcohol soluble extractives, foreign organic matter, moisture content and screening of the plant material for total microbial count (Total Bacterial Count and Total Fungal Count) in order to meet the most stringent quality regulations. Chromatographic finger printing of the herbs for their general profile or marker compounds using Thin Layer Chromatography will also form part of this study.

In another aspect of the present invention, extracts are isolated from fruits of the Lycopersicum esculentum, using conventional inorganic and organic solvent extraction and supercritical fluid extraction technology. Generally, extracts of the invention capable of functioning in a prophylactic or therapeutic manner as outlined herein can be extracted from any Lycopersicum esculentum plant, depending on the end purpose that is required of the extract.

In some of the embodiments of the present invention there is provided a process for preparing extracts of the invention from plant parts of Lycopersicum esculentum that comprises: a. Obtaining plant material from one or more parts of the plants.

b. Obtaining an extract from the plant material by contacting the plant material with
an aqueous, an ethanolic or an organic solvent, or a combination thereof,
optionally for a defined period of time thereby providing one or more plant
extracts.

c. Removing the plant material from the supernatant obtained in step b.

d. Optionally, lyophilizing said supernatant.

e. Analyzing the plant extracts for efficacy and presence of inhibitory activity
against cardiovascular disorders.

f. Selecting plant extracts having one or both of these activities.

The choice of selected plant material may be of any type but is preferably the fruits of the Lycopersicum esculentum plant.

The solvent extraction process may be selected from direct types such as extraction from plant parts in reflux extractor apparatus or in flasks at room temperature or at higher temperature with polar and/or non-polar solvent(s). Typically, the extraction process is as outlined herein. In another embodiment of the invention, the compositions for preventing, treating, or managing cardiovascular diseases and related disorders, comprises of direct composite extract of plant species with alcohol, water and hydroalcohol solvent and successive extract with solvents from non-polar to polar range. The compositions/medicaments may contain a pharmaceutically acceptable carrier, excipient, or diluent.

It will be apparent to the skilled addressee that the selection of solvent, or mixtures of solvents for each step in the isolation of extracts of the invention showing activity can be guided by results of bioassay analysis of separate fractions.

Some of the embodiments of the invention will describe the HPLC profiles and Mass spectrums of direct and successive solvent extracts of Lycopersicum esculentum plant parts thereby giving each extract an identity of itself.

The plants selected for the isolation of therapeutically relevant extracts/molecules to be used in the treatment of cardiovascular disorders, will be subjected to both targeted and non-targeted screening procedures. The ongoing-targeted screening procedures, which feature a comprehensive metabolite profiling of multitudes of phytoextracts, are envisaged in the study to facilitate the creation of a metabolite grid.

The successive extraction from plant extract will be carried out using soxhlet extractor. The solvents used, will be based on their sequential polarity starting from non-polar to polar, wherein, various classes of metabolites will be extracted viz. petroleum ether (phytosterols, fixed oils and fats), benzene (fixed oils and fats), chloroform (alkaloids), acetone (phytosterols, phenolics and tannins) ethanol (alkaloids, carbohydrates, glycosides, phytosterols, saponins, phenolics, tannins, proteins and amino acids) and water (alkaloids, carbohydrates, glycosides, saponins, phenolics, tannins, proteins, amino acids, gums and mucilage) at 65°C. These fractions will be lyophilized and stored in amber colored bottles at 4°C.

NOMENCLATUTRE OF THE EXTRACT ED:

AV-Avesthagen, D-Danone, HC-HDL cholesterol, plant genus and species,(temperature) of the extraction, solvent code(04-ethanol,08-water), (% of solvent used).

Phytochemical investigations will be also carried out on these extracts using various tests like Mayer's and Dagendorf s tests for alkaloids; Molisch, Fehling and Benedict tests for carbohydrates; Lieberman Buchard's test for phytosterols and triterpenes; spot test for fixed oils and fats; Ferric chloride and Lead acetate test for phenolic compounds and tannins; Ninhydrin and Biuret tests for protein and aminoacids; alcoholic precipitation followed by Molisch test for gum and mucilages.

The extracted fractions will be subjected to HPLC using u bondapak C i8 column (Waters Alliance 2695 Separation Module) to separate the constituent metabolites. The fractions are eluted using a combination (80:20, 60:40, 50:50, 40:60, 20:80) of methanol: water / acetonitrile: water. The gradient run will also be carried out wherever required. 5- lOul of sample is injected with flow rate of 1 ml/min and HPLC run will be performed for 30 minutes. The detection is done on photodiode array and the analysis of the results will be done with the help of Millennium™ software.

In some embodiments of the present invention, the analysis of output of LC-PDA and LC/MS/MS will be done using both LC-Solution and Analyst software of Applied Biosystems along with the script that is developed by Avesthagen to determine the molecular weight of chemical compounds by ionizing, separating and measuring molecular ions according to their mass-to-charge ratio (m/z).

The invention further describes the biotherapeutic potential of various extracts of Lycopersicum esculentum as described above, by studying their performance in cell based assay models.

In some of the embodiments of the present invention, mammalian cell based efficacy tests are conducted by growing Human hepatoblastoma cell line (Hep G2), in a flask with Eagle's Minimum Essential Medium (EMEM) containing 10% Fetal Bovine Serum (FBS), 1% glutamine-penicillin-streptomycin and 1% fungizone in a humidified incubator at 37°C in an atmosphere of 5% CO2 and 95% air. It is further subcultured when cell become 80% confluent they are subjected to treatment with bioactive under investigation. The incubation is followed by estimating levels of bio-markers for Cardiovascular disorder like apolipoprotein AI (Apo AI), total cholesterol, HDL cholesterol, triglyceride levels, Angiotensin converting enzyme (ACE), endothelial Nitric oxide synthase (eNOS), C-Reactive protein (CRP), Tumor Necrosis Factor (TNF-alpha),
Interleukins (IL-6) between the bioactive treated and untreated sets.

In another aspect of the invention there is provided a method for treating a disease in a mammal, which comprises administering to the said mammal an effective non-toxic amount of at least an extract from Lycopersicum esculentum as defined herein. Preferably the mammal is a human being. The skilled addressee will appreciate that "treating a disease" in a mammal means treating, that is to say, alleviating symptoms of the disease and may also mean managing a disease in the sense of preventing such a disease state either advancing i.e. getting worse or becoming more invasive, or slowing down the rate of advance of a disease.

The compositions/medicaments may contain a pharmaceutically acceptable carrier, excipient, or diluent. The compositions can be included as unit dosage suitable for parenteral, oral, or intravenous administration to a human. Alternatively, the compositions are dietary supplements, food compositions or beverage compositions suitable for human or animal consumption.

EXTRACTION OF Lycopersicum esculentum:

Extraction of Lycopersicum esculentum plant parts was carried out with alcohol, water and hydroalcohol solvent in reflux extractor apparatus or at room temperature under agitation followed by lyophilization under vacuum.

Example 1: Extraction Process A: Reflux Extraction

Powdered Lycopersicum esculentum plant material was weighed into the round bottom flask. Various concentrations of alcohol, water and hydro-alcohol was added in to the round-bottomed flask and placed on the mantle along with few (3-4) ceramic chips. The reflux condenser was then placed on the flask. Cold water was allowed to circulate continuously in the condenser from the tap. The mantle was switched on and the temperature was set to the boiling point of the solvent. The vapors of the solvent from the flask passed through the inlet of the extractor and condenses. The condensed (distilled) solvent thus extracts the compounds from it. This process is continuous as long as there is stable heat and water circulation. The extraction was continued for 2 hours at room temperature. After 2 hours the mantle was switched off and the water flow was stopped. After cooling the extract was collected separately and centrifuged.

The extract was concentrated by fitting the flask containing the extract with the empty soxhlet extractor body that in turn was fitted tightly with the water-cooled condenser. Continuous water flow was maintained and the flask was heated till the solvent from the flask was distilled and collected in the extractor body up to a level (One inch below the inlet). The temperature was reduced to avoid charring as the volume of the solvent reduced in the flask. The distilled solvent collected in the extractor was transferred to the solvent bottles and label appropriately. The process was continued till only very little solvent was left in the flask and no charring had occurred. Further concentration was done in the rotovapour apparatus to remove the solvent completely. The extract in the flask were swirled and were dried under vacuum. Storage and labeling of the extract was done to obtain the Extract ID.

EXAMPLE 2: PROCESS B ROOM TEMPERATURE EXTRACTION

Powdered Lycopersicum esculentum plant material was weighed into conical flask. Various concentrations of alcohol, water and hydro-alcohol was added in to the conical flask and placed under agitation at room temperature for 2 hrs. After 2 hrs, centrifuge at 4500 rpm for 15 minutes at 4°C. After centrifugation supernatant is taken and concentration is done in rotor evaporator. Further concentration is done in lyophilizer. The % yield of the extract obtained is calculated.

Calculations:

Calculate the percentage yield of the extract with respect to the initial weight of the plant material taken before extraction.

% Yield = wt. of lyophilized extract (after drying) * 100 Wt. of dry Plant material (initial)

EXAMPLE 3: EXTRACTION AVDHC27Le (25) 08(100):

100ml of the powdered plant material is taken in the round bottom flask and 1000ml 100% milli Q water is added to it. The extraction is started and continued for 2 hours at room temperature. After 2 hours of extraction, the solvent containing the extract is centrifuged at 4500 rpm for 15 minutes at 4°C temperature. The supernatant is taken and is concentrated in the rotor-vapour. Remaining drying and concentration of the extract is done by lyophilizing under vaccum. The dry weight of the extract is taken and the powdered dried extract is kept in an airtight glass container at room temperature.

Yield and Extracts characteristics: % of yield of extract is 26.0 and Beige colored powder.

EXAMPLE 4: PROCESS B EXTRACTION OF AVDHC27 Le (105) 08(100):
100ml of the powdered plant material is taken in the round bottom flask and 1000ml 100% milli Q water is added to it. The extraction is started and continued for 2 hours at boiling point of the solvent. After 2 hours of extraction, the solvent containing the extract is centrifuged at 4500 rpm for 15 minutes at 4°C temperature. The supernatant is taken and is concentrated in the rotor-vapour. Remaining drying and concentration of the extract is done by lyophilizing under vaccum. The dry weight of the extract is taken and the powdered dried extract is kept in an airtight glass container at room temperature.
Yield and Extracts characteristics: % of yield of extract is 29.2 and Beige colored powder.

HPLC FINGERPRINTING OF THE PHYTO EXTRACTS: EXAMPLE 5: HPLC FINGERPRINTING OF AVDHC27Le(25) 08(100):

Sample Preparation:

lml of HPLC grade water was added to l0mg of the sample and sonicated for 15mts.This was centrifuged at 13000rpm for lOmts.The supernatant was filtered using a Nylon filter (13mm x 0.2um) with a lml syringe into an injection vial.

2.Method:

Chromatogramic systems: Waters 2695 XE Separations Module equipped with auto sampler, degasser, PDA, detector, and Empower software. General system parameters:
Stroke volume (ul) - 50 ul (flow rates<= 1.23 ml/min), Bubble detector, Syringe draw rate (ul/sec) -Normal, Pre column volume (ul) - 0.0

Depth of needle (mm) - 0.6, Chart out - A%, Column position- no change, Needle Wash time-normal, Equilibration time - 20 min Degas: _Degas Mode - On, Events: Disabled,

Temperature: Disabled

Run Specifications:
Column Used: Atlantis, C18; 5um; 4.6X250mm
Solvent A - Acetonitrile (HPLC Grade)+ 0.1% TFA
Solvent B - Methanol (HPLC Grade)
Solvent C - Water (HPLC Grade)+ 0.1%TFA
Solvent D - Purge water (50%methanol)
Injection Volume: 20ul
Channel: Channel name - 2690/5 - System pressure
LC Time Program: Flow Rate: lml/min
Results and discussion:

The extract was injected at a constant volume of 25ul and its metabolite profiling monitored at 254nm. A comparative profiling was performed between the runs/ extract to check the reproducibility of the results.

Table 1: Peak information of AVDHC27Le (25) 08(100) extract at 254 nm.

Table 2: Summary of percentage conservation of AVDHC27Le (25) 08(100) extract scanned from 200 to 700 nm using Metagrid™ software
Constituents and conservation profiles of AVDHC27Le (25) 08(100) extract scanned from 200 to 700 nm using Metagrid™ software revealed that 30-50% conservation at the wavelength of 250/260/270/280/300/310 nm, 50-70% conservation at the wavelength of 210/220/230/240/290nm and 91-100% of conservation at 200/320/330/430/440/450/460/470/480/490/500/510/520/530 /540/550/560/570/580/590/600/610/620/630/640/650/660/670/680/690/700nm.

EXAMPLE 6: HPLC FINGERPRINTING OF AVDHC27Le (65) 08(100):

Sample Preparation:

20mg of the extract was weighed out into sterile eppendorf and 2ml of 50% methanol is added to it and sonicated for 15 minutes to ensure that the extract dissolves completely. The extract was centrifuged at 12,000rpm for 20 minutes at room temperature and the supernatant was filtered through 0.2u filters and collected into sterile recovery vials before analysis was performed.

Method:

Instrument parameters :_Shimadzu LC20AT

Data Acquisition: LC stop time: 55min, Acquisition time (PDA), Sampling: 3.125 Hz, Start

Time: 0 mts, End Time: 50mts, Time constant: 0.64sec.

Pump: Mode: Low Pressure Gradient, Total Pump flow: 1 ml/min, Solvent B Cone: 0 ml/min,

Solvent C Cone: 0 ml/min, Solvent D Conic: 0 ml/min, Maximum Pressure Limit: 5405
Controller: Model: CBM-20A, Power: On
Auto sampler: Model SEL 20A, Sample rack: 1.5ml standard, Rinsing volume: 200ul, Needle
stroke: 52mm, Control vial needle stroke: 52mm, Rinsing speed: 35ul/sec, Sampling speed:
15ul/sec, Purge time: 25 min, Rinse mode: Before and after aspiration, Rinse dip time: 10Sec.

Auto purge: Mobile Phase A: 5 min, Mobile Phase B: 5 min, Mobile Phase C: 5 min, Mobile Phase D: 5 min, Auto sampler: 5 min, Total Pump Flow: lml/min.

After checking all the above mentioned parameters the inlets from every solvent system were purged with the respective solvents for five minutes to clear the tubes of any air bubbles. Further the column was washed with HPLC grade water (100%) for 30mts and then equilibrated with the initial run conditions of the corresponding LC time program.

Run Specifications:

Column Used: Ascentis ™ RP -Amide,5um 25x4.6mm.
Solvent A: 0.1 %TFA in water
Solvent B: 0.1 %TFA in Acetonitrile
Injection Volume: 20ul

Run conditions: No of runs performed for each sample: 5,Injection volume: 25ul,
LC time program used: CVD2.MD17
LC Time Program:

Results and discussion:

The extract was injected at a constant volume of 25 u1 and its metabolite profiling monitored at 254nm. A comparative profiling was performed between the runs/ extract to check the reproducibility of the results.

Table 3: Peak information of AVDHC27Le (65) 08(100) extract at 254 nm.

Table 4: Summary of percentage conservation of AVDHC27Le (65) 08(100) extract scanned from 200 to 700 nm using Metagrid™ software.
Constituents and conservation profiles of AVDHC27Le (105) 08(100) extract scanned from 200 to 700 nm using Metagrid™ software revealed that 95-100% conservation at the wavelength Ofz200/210/230/240/250/260/270280/290/300/310V320/330/340/350/360/370/380/390/ 410/420/430/440/450/460/470/480/490/510/520/540/570/580/590/600/610/620/650/660/680/690/700nm, 90-95% conservation at the wavelength of 320/330/350 nm and 80-90% conservation at 360/370/390/400/500/530/550/560/630/640/670 nm.

CELL BASED ASSAY FOR APO Al ACTIVITY:

Lipids are transported throughout the body by complex structures called lipoproteins. Lipoproteins are classified into five major density classes: chylomicrons, very low-density lipoprotein (VLDL), intermediate density lipoprotein (IDL), low-density lipoprotein (LDL) and high-density lipoprotein (HDL). Associated with these lipoproteins, at least five major apolipoproteins have been described and have been labeled A through E. The principle apolipoproteins of HDL are the A apolipoproteins, constituting nearly 90% of the protein mass. Thus, apoA-I has been found to be more powerful as a marker in some studies for coronary disease than other HDL components (eg. cholesterol). Apo A-I enhances fibrinolysis and antiplatelet activity, thus possibly having an antithrombotic role in coronary disease prevention. Studies have indicated that the increased apoA-I gene expression in apoE-deficient mice markedly suppressed arteriosclerosis, further supporting a protective role for apoA-I.

The composite extracts were taken forward for efficacy validation using HepG2 cells based assay system. This step is for estimating bioactivity of the composite extracts to modulate levels of apoAI.

ApoAl a major protein of HDL, initiates cholesterol efflux and thereby facilitates removal of excess tissue (eg. arterial) cholesterol by the process of reverse cholesterol transport.

Human hepatoblastoma cell line (Hep G2) was grown in a T-75 flasks with 15 ml of EMEM containing 10% FBS, 1% glutamine-penicillin-streptomycin and 1% fungizone in a humidified incubator at 37°C in an atmosphere of 5% CO2 and 95% air. Subcultures when cell become 80% confluent by trypsinization with PBS containing 0.5 mmol/L EDTA.

The cells were plated in 96 well plates at a concentration of 3x106 cells/ml and grown until they attained 75% to 80% confluence. A dose response curve of gemfibrozil (0 to 400 umol/L) / herbal extracts on apoA-I secretion for varying times (24 hours) was prepared and at the termination of the incubation, 50-uL of cell supernatant was taken to estimate the apoA-1 concentration by an enzyme-linked immunosorbent assay (ELISA)
After termination of the reaction, the cell supernatant was removed and washed twice with PBS and the cells were lysed either with 100 uL of 1% Triton X 100 or with 0.1% SDS. 10 ul of the sample was taken and added to 250 ul of Bradford reagent and the reading was taken at 590 nm.

Calculation was done according to the kit protocol as ng /ug of cellular protein and the graphs were generated using Graphpedprism Software.

EXAMPLE 7: APO Al ANALYSIS OF AVDHC27 Le (25)08(100):

AVDHC27Le (25) 08 (100) water extract RT did not show any cytotoxic effect on HepG2 cells at all the studied doses (0.1 to 500 ug/ml) at 24 hrs of treatment as shown in the figure 7

AVDHC27Le (25) 08 (100) water extract RT exhibited an increase in ApoAl levels at all the studied doses of 0.1 - 500 ug/ml. However maximum increase was observed at the lower doses of 0.1-10 ug/ml showing 1.3-1.5 fold increase when compared to that of the positive control (Gemfibrozil) exhibiting only 1.3 fold increase. The same demonstrated 1.3-fold increase at the higher doses of 100-500 fig/ml. The result is described in the figure 8.

Thus the result seems to suggest that AVDHC27Le (25) 08 (100) water extract RT has high efficacy and potency in elevating the level of ApoAl in human hepatoblastoma cell line (HepG2).

Table 5: Effect of AVDHC27Le (25) 08 (100) water extract RT on ApoAl levels in HepG2 (Human hepatoma) cells at 24 hrs of treatment.

- Extract was diluted in water, sterilized though a 0.22 u filter and used for the assay. The experiment was terminated at 24 hrs.

** - Results are the mean ± SD of triplicates per sample. AVDHC27Le (25) 08 (100) water extract RT is compared to the cell control while Gemfibrozil has been compared respective

Ethanol control.
***-P<0.0001, **-P< 0.001

EXAMPLE 8: APO Al ANALYSIS OF AVDHC27 Le (65) 08(100):

AVDHC27Le (65) 08 (100) water extract reflux did not show any cytotoxic effect on HepG2 cells at all the studied doses (0.1 to 500 ug/mi) at 24 hrs of treatment, as shown in the figure 9. AVDHC27Le (65) 08 (100) water extract reflux exhibited an increase in ApoAl levels at all the studied doses of 0.1 - 500 ug/ml. Maximum increase was observed at the lowest dose of 0.1 ug/ml showing 1.4 fold increase when compared to that of the positive control (Gemfibrozil) exhibiting only 1.3 fold increase. However the same exhibited 1.2-1.3 fold increase at all the higher doses of 1-500 ug/ml. Thus the result, shown in the figurelO seems to suggest that AVDHC27Le (65) 08 (100) water extract reflux has high efficacy and potency in elevating the level of ApoAl in human hepatoblastoma cell line (HepG2).

Table 6: Effect of AVDHC27Le (65) 08 (100) water extract reflux on ApoAl levels in HepG2 (Human hepatoblastoma) cells at 24 hrs of treatment.

* - The sample was diluted in water, sterilized though a 0.22u filter and used for the assay. The experiment was terminated at 24 hrs.

** - Results are the mean ± SD of triplicates per sample. AVDHC27Le (65) 08 (100) water extract reflux is compared to the cell control while Gemfibrozil has been compared respective

Ethanol control.

**-P<0.01,*-P<0.05

EXAMPLE 9: MODES OF ADMINISTRATION:

For administration to a mammal, the therapeutic composition can be formulated as a pharmaceutical or naturopathic formulation such as phytoceuticals or nutraceuticals, for oral, topical, rectal or parenteral administration or for administration by inhalation or spray. The phytoceutical or naturopathic formulation may comprise the one or more plant extracts in dosage unit formulations containing the conventional non-toxic physiologically acceptable carriers, adjuvants and vehicles. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrathecal, intrasternal injections or infusion techniques.

The pharmaceutical or naturopathic formulations may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion hard or soft capsules, or syrups or elixirs. The therapeutic compositions of the invention may be formulated as phytoceuticals, or nutraceuticals.

Phytoceuticals may optionally comprise other plant-derived components and can therefore be delivered by such non-limiting vehicles as teas, tonics, juices or syrups. Nutraceuticals contemplated by the present invention may provide nutritional and/or supplemental benefits and therefore be delivered, for example as foods, dietary supplements, extracts, beverages or the like. Phytoceutical and nutraceuticals can be administered in accordance with conventional treatment programs and/or may be a part of the dietary or supplemental program.

Formulations intended for oral use may be prepared according to methods known in art for the manufacture of pharmaceutical compositions and may contain one or more agents selected from the group of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide palatable preparations.

Tablets contain the active ingredient in admixture with suitable non-toxic physiologically acceptable excipients including, for example, inert diluents, such as calcium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as corn starch, or alginic acid, binding agents, such as starch, gelatine or acacia, and lubricating agents, such as magnesium stearate, stearic acid or talc. The tablets can be uncoated, or they may be coated by known techniques in order to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period.

Various additives or carriers can be incorporated into the orally delivered pharmaceutical naturopathic formulations or the invention. Optional additives of the present composition include, without limitation, phospholipids, such as phosphatidyl glycerol, phosphotidyl inositol, phosphotidyl serine, phosphotidyl choline, phosphotidyl ethanolamine as well as phosphatidic acids, ceramide, cerebrosides, sphingomyelins and cardiolipins.

Pharmaceutical or naturopathic formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatine capsules wherein the active ingredient is mixed with water or an oil based medium such as peanut oil, liquid paraffin or olive oil.

A syrup may be made by adding the active extract to a concentrated, aqueous solution of a sugar, for example sucrose, to which may also be added any necessary ingredients. Such accessory ingredient (s) may include flavorings, an agent to retard crystallisation of the sugar or an agent to increase the solubility of any other ingredients, such as polyhydric alcohol for example glycerol or sorbitol.

Oily suspensions may be formulated by suspending the plant extract(s) in a vegetable oil, for example, arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or acetyl alcohol. Sweetening agents and/or flavoring agents may be added to provide palatable oral preparations. These formulations can be preserved by the addition of an anti¬oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation suitable for an aqueous suspension by the addition of water provide the active ingredient in admixture with dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents,sweetening, flavoring and coloring agents may also be present.

In a further aspect of the invention there is provided a comestible, that is to say, a foodtuff comprising at least an extract of the invention, typically in dried form, such as in a lyophilised form. The skilled addressee will appreciate that such cosmetibles may contain more than one extract of the invention and may be used. Such foodstuffs may be used in a prophylactic manner and may contain further extracts having a similar function to the first added extract or further added extracts may be added that have a different prophylactic function. Thus a foodstuff could either comprise extracts that provide for a comestible having a single functional aspect, or a comestible may have a multi-functional prophylactic effect against two or more disease types. It is thought that a multi-functional role could be assigned to pharmaceutical formulations comprising two or more extracts possessing dissimilar therapeutic or prophylactic properties desgined either for prophylaxis or for the treatment of more than one disease(s) in a mammal, particularly in a human.

The type of foodstuff or comestible to which at least an extract of the invention may be added includes any processed food such as confectionaries, baked products including breads such as loafs, and flat breads such as pitta bread, naan bread and the like, cakes, snack foods such as muesli bars, compressed dried fruit bars, biscuits, dairy products such as yoghurts, milk and milk-based products such as custards, cream, cheese, butter and creme fraiche, simulated dairy food product such as Elmlea products, fruits and vegetable juices, water, aerated drinks, such as carbonated soft drinks and non-aerated drinks such as squashes, soya milk, rice milk and coconut milk and the like, pastas, noodles, vegetables, seed and nut oils, fruited oils such as sunflower oil, rapeseed oil, olive oil, walnut, hazelnut, and sesame seed oil and the like, and frozen confectionaries such as ice cream, iced yoghurts and the like.

We Claim:

1. A method of obtaining a plant extract from the plant capable of delaying the onset and/or management of cardiovascular disorders comprising the steps:

a. Obtaining plant material from one or more parts of the plants of claim 1.

b. Obtaining an extract from the plant material by contacting the plant material with an
aqueous, an ethanolic or an organic solvent, or a combination thereof, optionally for a
defined period of time thereby providing one or more plant extracts.

c. Removing the plant material from the supernatant obtained in step b.

d. Optionally, lyophilizing said supernatant.

e. Analyzing the plant extracts for efficacy and presence of inhibitory activity against cardiovascular disorders, such as herein described.

f. Selecting plant extracts having one or both of these activities.

2. A method according to claim 1 wherein the plant extracts is derived from the plant of the genus Lycopersicum.

3.A method according to claim 1, wherein, the plant extracts are extracted from leaves, seeds, roots, stems, flowers, or various combinations thereof but preferably from the fruit of the plant.

4.The plant extracts AVDHC27Le(25)08(100), AVDHC27Le(65)08(100) produced by the process of claim 1.

5. A method of profiling plant extract(s) produced by the process of claim 1, comprising steps of:

(a) Preparing sample solution of plant part(s); (b) injecting the sample solution into HPLC column; (c) detecting eluant(s) from the HPLC column; and (d) analyzing the eluant(s) via visual interface to profile the plant extract(s).

6.The method as claimed in claim 5, wherein said profiling involves analyzing chromatographic data, assessing efficacy and bioactivity screening of plant extracts).

7. The method as claimed in claim 5, wherein the HPLC column is maintained at a temperature of about 37 °C.

8. The method as claimed in claim 5, wherein said HPLC is carried by gradient technique using water, methanol and acetonitrile as mobile phase solvents.

9.The method as claimed in claim 6, wherein the chromatographic data is obtained after every second from wavelength ranging from 190 nm to 700 nm with a resolution of about 1.2 nm.

l0.The method as claimed in claim 5, wherein said HPLC comprises reverse phase liquid
chromatography coupled with photo diode array detector or reverse phase liquid chromatography
and photo diode array detector along with mass spectrometer.

11 .The method as claimed in claim 5, wherein said visual interface allows user to pictorially visualize differences in chromatographic profile of an extract and provides unique and consistent chromatographic peaks.

12. Visual interface for displaying plant extract analysis having predetermined colored data points for depicting retention time and tolerance range as herein described along with accompanying examples and drawings.

13. A method of treating Cardiovascular disease and/or related conditions thereof in a mammal in need thereof, said method comprising step of administering pharmaceutically acceptable amount of Lycopersicum plant extract, optionally along with pharmaceutically acceptable additives, to the said mammal.

14. The method of treating as claimed in claim 13, wherein the lycopersicum plant extract is having triglycerides or cholesterol decreasing activity or enhancing HDL activity, and alpha glucosidase activity.

15. An herbal composition for therapeutic use, the composition comprising extracts of Lycopersicum esculentum species.

16. A composition of claim 15, comprising one or more plant extracts, wherein the extracts can be formulated as phytoceuticals or nutraceuticals not limiting to teas, tonics, juices, syrups and nutraceuticals providing nutritional benefits in the form of foods, beverages, supplements and the like.

17. A composition of claim 15, comprising one or more plant extracts, wherein the pharmaceutical or naturopathic formulations may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion hard or soft capsules, or syrups or elixirs.

18. The composition of claim 15, wherein the pharmaceutically acceptable additives are selected from a group consisting of granulating agents, binding agents, lubricating agents, disintegrating agents, sweetening agents, coloring agents, flavoring agents, coating agents, plasticizers, preservatives, suspending agents, emulsifying agents, and spheronization agents.

19. An herbal composition comprising of Citrus plant extract, the method of treating
cardiovascular disorders substantially as herein described along with accompanying examples and figures.