NOVEL COMBINATIONS WITH HORSEGRAM FOR THE MANAGEMENT OF OBESITY
FIELD OF INVENTION
The present invention relates to the field of anti=obesity agents and to a combination therapy using the combination of statins and legumes and administrated separately or in combination and preferably as a single fixed bi-combination product. The invention is further related to use of these agents in a combination therapy for the treatment, the synergistic effects they pfödücé för thé pfëvëntiöh öï cöntföl öf obesity and variöüs öthër assÖciatéd pathölógiës. Additionally, the invention is related to the reduced usage of statins in this combination therapy, when compared with the normal levels of usage when administrated alone to achieve the same or substantiaÜy simiJar results.
BACKGROUND OF INVENTION
The epidemie of obesity has skyrocketed in the recent years in the western countries and the dëvëlöping cöüntriës. The major óbësity fëlatëd disorders aré attfibütëd tö thé liféstyle changes due to rapid urbanisation and modernisation. The pandemic effect of an obese individual is detrimental, and it may also cause other related complications such as CVD, type II diabetes, cancer, hypertension and dyslipidemia. The aetiology of obesity is ascribed to multiple factors such as genetic, endocrine, gut microbiome, environmental and behavioural factors that results in health-related complications such as metabolic disorders, gastrointestina 1 disorders, cardiovascular disorders, respiratory disorders and malignancies.
The main goal in obesity management is to reduce excess weight and maintain it for a longer period and various modes of management techniques have been made such as diet, physical activity, behavioural therapy, pharmacotherapy and surgery and the use of such techniques in an individual either alone or as a combination therapy depends on the individual weight, co-morbidities and expected targets.
Among these, pharmacotherapy involves a variety of different classes of drugs that can aid in obesity management based on chemical nature and its mechanism of action. The drugs used in pharmacotherapy can be further classified as metabolic inhibitors, appetite suppressants,

hypolipidemic agents, hypoglycaemic agents, antioxidant agents, anti-inflammatory agents and lipid reguïating agents. Lipids are the major biomolecules that helps in transferring energy from food to body ceils. Cholesterol and triglycerides play an active part in the normal body function and elevated level of üpids can results in detrimental effects.
Currently, two groups of drugs namely statins and fibrates are used for lowering the lipid lëvëls. Stairis sübdüé cholesterol Öiösyrithésis by ihhiBitirïg HMG-CöA fédüctasë. Statins that are currently prescribed for the treatment of hyperlipidaemia and/or hypercholesterolemia comprises atorvastatin (Lipitor® from Pfizer), simvastatin (Zocor® from Merck), pravastatin (Pravachol® from Bristol Myers Squibb), fluvastatin (Lescol® from Novartis), lovastatin (Mevacor® from Merck), and rosuvastatin (Crestor® from AstraZeneca). The present day hyperlipidemic patients take both the allopathie medicines as well as the dietary ingredients which may or may not exhibit spme fopd-drug interactions. Hence, the present study was taken up to study the positive effects of these legumes and the influence of these legumes with atorvastatin in diet induced obesity and on the risk factors of CVD.
It is therefore an object of the invention to provide a method of enhancing the effectiveness of obesity management by administering to a patiënt in need thereof co-therapy which includes at least one statin and dietary materials. It is another object of the invention to provide a cöfïïpósitión córiïprisitig' at least öhe statiri, and at least ónë fóöd mater ia I sëlëcted fröfh thé group of legumes. It is still another object of the invention to provide a synergistic composition comprising at least one statin and legumes. A further objective of the invention is to provide a statin/ legumes anti-obesity co-therapy where the statin is atorvastatin or a pharmaceuticaiïy acceptable salt thereof and the legume is Horse gram.
SUMMARY OF THE INVENTION
The above discussed and other objects of the invention can be achieved in subjects in nëed of cholesterol and/or weight rëdüctióh ör cöritröl By way of ëómBihatiön thëfapy tfcatmënt with at least one statin and at least one dietary legume. The statin used in the combination therapy for its synergistic effects in the treatment of obesity is atorvastatin or a pharmaceuticaiïy acceptable salt thereof and the legume is Horse gram.
The other objectives and advantages of the present disciosure wiü be further appreciated and understood when considered in combination with the foHowing description, tables and

accompanying drawings. While the following description may contain specific details describing particular embodiments of the present disclosure, this should not be construed as limitations to the scope of the present disclosure but rather as an exemplification of preferable embodiments. For each aspect of the present disclosure, many variations are possible as suggested herein that are known to those of ordinary skill in the art. A variety of changes and mödificatiöns may bé made within Üië scope of thé present disclosure without departirig fröm the spirit thereof
BRIEF DESCRIPTION OF THE DRAWING
FIG. la and lb are the representation for the effect of Horsegram on body weight before and after the treatment respectively.
FIG. 2 (a-d) are the representative images for the effect of Horsegram on Lipid Profiles a. Triglyceride, b. Cholesterol, c. HDL and d. VLÖL respectively.
FIG. 3 (a-d) are the representative images for the effect of Horsegram on Renal Markers a. Uréa, b. Cféatihiné, c. CRP and d. TNFci respectively.
FIG. 4 (a -f) are the representative images for the effect of Horsegram on Relative Gene Expression of Lipid Metabolizing proteins in Hepatic Tissue, a. SREBP-1, b. FAS,
c. ACC, d. LPL, e. FABP and f. HMG CoA
FÏG. 5 (a-d) are the representative images for the effect of Horsegram on Relative Gene Expression of Adipogenesis in Adipose Tissue a. Leptin, b. Adiponectin, c.UCP-2 and
d. Resistin respectively.
FIG. 6 (a-d) are the representative images for the effect of Horsegram on Relative Gene Expression on Inflammatory Markers in Adipose and Hepatic Tissues a. NF-kB (Adipose Tissue), b. IL6 (Adipose Tissue), c. NF-kB (Liver Tissue) and d. IL6 (Liver Tissue).
FIG. 7 (a-e) are the representative images showing the histopathological changes in the Liver Tissue
FIG. 8 (a-e) are the representative images showing the histopathological changes in the Brown Fat Tissue

DETAILED DESCRIPTION OF THE INVENTION
the invention aims to solve the technical problem of improving cholesterol and/or weight reductiöh of cöhtról by way öf cömbihatiöh thêfapy treatment with at least one statin and at least one dietary legume. Provided herein are the compositions, combinations and other possible additions to the compositions that can be used in the treatment of obesity and its related pathologies.
The dosage of the combinati on therapy of this invention can be separate, simultaneous or at a sequence and in addition to the two agents mentioned above, may include other optional additional agents. The combination of statins and legumes (horse gram) will be in different dosage forms and may be administered in an amount in the ranges from below the minimal amount to provide the synergistic results as proposed in this invention to the maximum ranges as appropriatc. Similarly, the additional optional actives can also be added in preferred dosages with no set ratio of one component to the other in the composition. Any reference to compounds in this invention is to be understood to be comprising it's solvates, hydrates and its afthydföüs fbrm.
Statins and fibrates play important role in lipid lowering and are the general preferred hypolipidemic agents. The statins in general subdue the cholesterol biosynthesis by inhibiting HMG-CoA reductase. The statins possess a pleiotropic role such as inhibition of cholesterol biosynthesis, hamper the actlvation of smalï G proteins by inhibiting isoprenoids synthesis, suppress the proinflammatory molecules, inhibit reactive oxygen species production and inhibit migration and proliferation of smooth muscle cells.
Statins that are currently prescribed for the treatment of hyperlipidaemia and/or
hyperehölëstëfölémia cömpfisës atörvastatiri (Lipitöf® frörri Pfizer), sihïvastatiri (Zöcöf®
from Merck), pravastatin (Pravachol® from Bristol Myers Squibb), fluvastatin (Lescol®
from Novartis), lovastatin (Mevacor® from Merck), and rosuvastatin (Crestor® from
AstraZeneca). The statin selected for use in the present invention as hypolipidemic agent is
atörvastatiri.
Legumes are rich in nutrients with lesser energy density. The commonly consumed legumes include alfalfa, dover, lupin, green beans and peas, peanuts, soybeans, dry beans, broad beans, dry peas, chickpeas, ground nuts, horse gram and lentiïs. The iegumes contain

phytochemicals including enzyme inhibitors, phytohaemagglutinins (lectins), phytoestrogens, oligosaccharides, saponins and phenolic compounds. Hence, increased legume consumption prevents obesity and thereby controls coronary heart disease, diabetes and metabolic syndrome owing to its phytochemicals. The legume selected for use in the present invention is horse gram that will be used with the hypolipidemic agent, atorvastatin.
Binomially known as Horse gram (Macrotyioma uniflorum) is highly nutritious and known for its ethnomedicinal values. It is rich in protein, carbohydrate, iron, manganese etc. Horse gram is the störehöusë of protëirïs. It is primarily fich in lysiné content and öthër aminö acids like arginine, histidine, valine, leucine etc, are also present in it. It has limited amounts of tryptophan and methionine. Horse gram is rich in both saturated and unsaturated fatty acids. Presence of a high amount of unsaturated fatty acids helps the human body to maintain cell membrane formation.
Horse gram possesses a plenty of medicinal benefits in it. Most of them are contributed by the presence of polyphenols — plant's secondary metabolites. It contains an array of bioactive molecules such as phenolic acids (caffeic acid, chlorogenic acid, ferulic acid, p-coumaric acid, siriapic acid, gallic acid), flavöhöids (daidzëih, gënistein, kaërripfërol, myricëtih, quercetin) and Anthocyanins (Cyanidin, delphinidin, malvidin, petunidin).
In-vivo studies are conducted to reveal the inherent properties of legumes and its combined effect with atorvastatin in mitigating the pathophysiological changes in obesity. The combinational therapy of the invention with atorvastatin and Horse gram would yield a proper scientific investigation and authentication of the use of legumes and its combined effect with atorvastatin in weight management. The synergistic effect of the combinational therapy is explained in detail with the help of the following study as one of the possible ëmbódimëfits of this ihvëhtiön.
Subsequently, the combined effect of legumes and atorvastatin, a hypolipidemic drug over obesity in experimental animal model was studied. The obese conditions in the experimental rodent models were induced by high fat diet for a period of 12 weeks. The animals were groüped and treated with individüal test drugs (HG and ATO) arid cömbiriatiön öf test drugs (HG+ATO); The effective dose was fixed as 300 mg/kg for legumes after in vitro studies and after 28 days of oral administration of test drugs, various biochemical, systemic and moiecular parameters were analyzed.

The anthropometric parameters such as body weight, feed and water intake, organ and fat weight were measured. The maximum reduction in body weight was observed in the GN+ATO group. With reference to feed intake* the combination of GN+ATO showed significant reduction in the feed intake when compared to the other groups. Subsequently, there was a significant increase in the weight of the liver in HFD fed rats compared to the normal control which might be due to the accumulation of lipids in the organ. There were no significant treatment related changes in the organ weight of vital organs such as heart, kidney and braih. It is nótëworthy to ideritiry that hórse gram ihdividüally arid in combination with ATO (HG +ATO) significantly resisted the increase in fat mass gain.
Obesity is one of the major leading risk factors for cardiovascular disease (CVD) which mediates dyslipidemia, hypertension and insulin resistance. Hence, the blood pressure and heart rate were measured before and after the treatment. Moderate changes in the systolic and diastolic blood pressure were observed, though not significant. Legume intake stabilized the heart rate.
Across the treated groups, the glucose levels were significantly reduced in all treatment groups. The resistance to insulin were high across all the groups, after eight weeks of high fat diet and the insulin resistance declined in the treated groups. Analogous to the effect on serum glucose levels, significant improvement over insulin sensitivity (as evidenced by increase in insulin levels and HOMA IR) were observed in combination treated groups. The TG, cholesterol and VLDL levels were greatly reduced arid HDL level was increased in ATO and other groups after dosing as observed on the 28th day.
The markers of renal function - urea and creatinine was significantly eievated in HFD group compared to normal diet group and there were no significant changes in all the treated groups. Likewise, CRP level was greatly decreased in all the treated groups but (HG+ATÖ) showed marked difference. However, there was no changes in TNF-a levels. L
Liver function tests which incïudes the activity of ALT, AST, ALP, y-GT, LDH and bilirubin were performed to confirm the degree of liver damage. Overall (HG+ATO) group showed effective protêction agairist liver damage. Thé liver plays a uriiqüé rölé iri cöritrolling carbohydrate metabolism by maintaining glucose level. Treatment with legumes improved hepatic hexokinase activity which can cause increased glycolysis and increased utilization of glucose for energy production and in turn stimulate the activity of these enzymes and pyruvate level. Likewise, administration of both the legumes significantly restored glucose-^?

phosphatase and fructose-1, 6-bisphosphatase activity in the HFD fed rats. It might be due to the activation of hexokinase enzyme or decrease in gluconeogenetic enzymes by promoting the uptake of glucose and storage in tissues.
Öxidative stress plays a prodigious role in the development of metabolic disorders. In the present study both enzymatic and non-enzymatic antioxidants such as catalase, GST, SOD, GSH and TBARS were analyzed in the major organs like liver, heart and kidney. It is interesting to note that the combination group specifically (HG+ATO) reduced the öxidative stress in all the vital organs compared to the atorvastatin alone treated group.
In the case of lysosomal hydrolases, HFD fed rats showed marked elevation in cathepsin D and MPO in the liver, heart and kidhëy tissues whén compared tó thé riornïaJ fats. Hórsè gram and its combination with ATO showed decrease in both the enzyme levels substantiating the potential beneficia! role of horse gram in mitigating inflammation pathologies in obesity.
To elucidate the possible mechanism of action of the food supplements, gene expression in the liver and adipose tissue and protein expression in the liver and heart was studied. Impaired lipid metabolism inhibits normal lipolysis process and, thus, leads to increased cell number and larger mass of adipose tissue. The key lipid metabolism regulating genes were studied in the hepatic tissues wherein SREBP, ACC, LPL and FABP exhibited decreased expfessiöh Ievel in the cómbihatioh gïóüps (HG+ATO & GN+ATO). However, the expression of HMGCoA reductase and FAS were better in individually treated legumes (Horse gram) as compared to the combination groups.
To explore the mechanism of underlying suppression of adipogenesis by legumes and its combination with ATO in modulating obesity associated conditions, the mRNA levels of genes such as resistin, leptin, adiponectin and UCP-2 were examined in adipose tissues. Herein, Horse gram individually as well as its combination with ATO (HG+ATO) could substantially reverse the expression of the respective genes.
Likewise, inflammation play major pathological role in obesity. HFD fed rats showed marked elevation in NF-kB and IL-2 in the liver and adipose tissue. The expression was better in HG and its combination with ATO treated rats. In case of protein 139 expression, we examined the specifie markers which are involved in the pathology of obesity. During the pathogenesis óf obesity, the major orgari such as liver and heart airé affected the most. Thüs, protein expression of iiver and heart was studied in the present study and markers such as AMPK,

PPAR-a and PPAR-y for lipid metabolism, HO-1 for oxidative stress and NF-KB for inflammation was checked.
In the present study, there was a marked improvement in the protein expression of these markers in the HG and its combination with ATO (HG+ATO) treated groups. This was in congruent to the gene expression studies. The gene and protein expression studies reveal that the amelioration of obesity and its associated disease pathoiogies was achieved by action on the inflammation, oxidative stress, adipogenesis and lipid related signalling pathways.
The histopathoiogical results also confirm the pathological changes in the experimental animals and substantiates the influence of legumes and their combination with atorvastatin in organs such as liver, heart, kidney and brown fat. In the liver, highest protection against this lesion was observed in ATO group. Among the test drug groups, almost equal protection was observed with HG group. In the ATO combination group, the protection level was further better, but less than ATO alone group. In the heart, occasional fatty changes in cardiomyocytes was observed in all the groups. No significant difference was observed between the groups. Likewise, in the kidneys, medulla, cortex, glomeruli and tubules appeared normal in all the groups. In the brown fat, it is interesting to note that varied levels of protection from whitening of brown fat was observed in HG group with or without ATO. However, ATO alone group showed highest level of whitening of brown fat compared to all other groups confirming the major side effect of atorvastatin.
Experimental Studies
The following studies were performed in the test group (rodents) to understand better the in- vivo hypoüpidemic effect of legumes and its combination with atorvastatin and its synergistic effect against high fat diet induced obesity; Furtheri in order to identify the potential influence of these legumes on obesity induced metaboiic syndrome the various physiological, biochemical, molecular and histopathoiogical changes were recorded and discussed below. The amount of food intake ranges was optimised using experimental models.
Effect of horse gram on Body Weïght
The group of rodents undergoing this treatment were classified basically in to five groups based on their dietary input and further classified into groups such as Normal Control, HFD, ATO, HG and the combination group HG + ATO. Figures IA and 1B are the representation for the observation on the effect of horse gram on bodyweight beföre and after the treatment.

From the figure IA, it is clear that, due to consumption of high fat diet the anima Is became overweight and obese as evidenced by the increase in body weight.
Similarly, after the treatment HFD fed animals co-treated with horsegram and ATO significantly maintained the body weight nearer to Normal diet fed rats. But in this case, the maximum reduction in body weight was observed in the combination group of HG + ATO.
Figures 2A-2D shows the effect of horsegram on lipid pro file. Values are expressed in Mean-SEM, n=7 animals/group, Statistical analysis was performed using one way ANOVA foliowed by Tukey's multiple comparison test, #, ## and### indicates p value< 0.05, 0.01 and 0.001 respectively Vs Normal Control; *,**and *** indicates p value < 0.05,0.01 and 0.001 respectively Vs HFD group.
From the figures 2A to 2D, it is clear that, there is a significant decrease in triglycerides, cholesterols and VLDL was observed in horsegram and HG+ATO combination. The combination also enhanced the HDL level which is a good cholesterol thereby showing its anti-hyperiipidemic property.
Figures 3A-3D shows the effect of horsegram on Renal markers. Values are expressed in Mean-SEM, n=4 animals/group, Statistical analysis was performed using one way ANOVA foliowed by Tukey's multiple comparison test, ## and ### indicates p value< 0.01 and 0.001 respectively Vs Normal Control **ahd **-* indicates p valüe < 0.01 and 0.001 respectively Vs HFD group.
From the figures 3A to 3D, it can be observed that, Creatinine and urea nitrogen are constantly excreted through the kidneys. Elevated levels suggest kidney malfunction in observed in obese conditions. Although there is decreased urea, creatinine levels in HG+ATO treated rats compared to obese rats, it is not significant. Also, with regards to the renal ftinction markers associated with inflammation, the combination group reduces the C-reactive protein which is a marker for inflammation.
Figures 4A-4F shows the effect of horsegram on lipid metabolizing proteins in hepatic tissue. Values are expressed in Mean-SEM, n=4 animals/group, Statistical analysis was performed using one way ANOVA foliowed by Tukey's multiple comparison test, # indicates p value< 0.05 Vs Normal Control *, **and *** indicates p value < 0.05 and 0.01 and 0.001 respectively Vs HFD groüps.

From the fïgures 4A to 4F, it can be inferred that the impaired lipid metabolism inhibits normal lipolysis process. Being the major form stored in adipocytes and liver, excessive serum fatty acids and cholesterol content leads to enlargement of storage cells and^ thus» the expression of respective genes that control their biosynthesis were evaluated.
Sterol regulatory element binding protein 1 (SREBP- 1) is a key lipogenic transcription factor, which directly activates the expression of many genes (including FAS (Fatty acid synthase), ACC (Acetyl-CoA carboxylase)), dedicated to the synthesis and uptake of fatty acids, cholesterol, and triglycéridë. All thésé ttirëe genes are reïative dëcreasëd in the combination groups when compared to HFD fed group;
HMG-CoA reductase (3-hydroxy-3-methyi-glutaryi-coenzyme A reductase) is most abundantly expressed in the liver and plays a central role in the regulation of plasma cholesterol concentration as it is the rate limiting enzyme in cholesterol biosynthesis. Statins are a major class of drug which is effective in lowering cholesterol levels and a potent inhibitor of HMGCoA reductase. Significant decrease in the protein expression was observed in the treatment groups which confirms that the cholesterol metabolism is modulated.
Figures 5A-5F shows the effect of horsegram on reïative gene expression of Adipogenesis in adipose tissue. Values are expressed in Mean-SEM, n=4 animals/group, Statistical analysis was performed using one way ANOVA foliowed by Tukey's multiple comparison test, # indicates p value< 0.05 Vs Normal Control * and ** indicates p value < 0.05 and 0.01 rëspëctively Vs HFD gïóüp.
From the figures 5A to 5F, it can be inferred that HG + ATO combination downregulatcs the expression of ieptin and resistin and upregulates the expression of adiponectin and UOP-2 (uncoupling protein-2), The anti-obese effect was profound in the combination groups due to moduiation of these adipokine genes.
Figures 6A-6F shows the effect of horsegram on reïative gene expression of Inflammatory markers in adipose and hepatic tissue. Values are expressed in Mean-SEM, n=4 animals/group, Statistical analysis was performed using one way ANOVA foliowed by Tukéy's multiple comparison test, # and ## indicates p valüë< O.OSahd 0.01 rëspëctively Vs Normal Control * and *** indicates p value < 0.05 and 0.001 rëspëctively Vs HFD group.

From the figures 6A to 6F, it can be inferred that the inflammation plays a major pathological role in obesity. In diseased condition, the inflammatory markers are elevated, on treatment with horsegram and its combination the gene expression is significantly reduced. Thus, the reduction of inflammatory molecules through combination supports that the therapeutic effect of horsegram and its combination may act as a promising therapeutic agent for inflammation which is the most important pathology in most of the metabolic disorders such as cardiovascular disease and diabetes.
Figure 7 shows the histopathologicaj changes in Liver Tissue. Liver tissue stained with haematoxylin and eosin (Magnification =200X). Arrows and * denotes, in normal hepatocytes and hepatic cord (arrow) and portal area (■*). In HFD, diffuse micro vesicular fatty change (*). Ballooning of cells, shrunken or degenerating nucleus (arrow) observed in focal areas. ATO group shows hepatocytes and hepatic cords appear normal except occasional fatty changes (arrow), HG shows moderate diffuse micro vesicular fatty changes in hepatocytes. Occasional macro vesicular fatty change with displaced compressed nucleus (arrow). In, HG + ATO, minimal diffuse micro vesicular fatty changes in hepatocytes.
From the figure 7, it can be inferred that the liver showed diffuse micro vesicular fatty changes observed in liver parenchyma of high fat group with one or two fat vacuoles per hepatócytë. Also, focal areas of rhicró vesicular fatty change having multiple fat vacüólés per hepatocyte with ballooning of cells, shrinkage or loss of nucleus was observed. Occasionally, focal areas of necrosis were also observed. Highest protection against this lesiön was observed in ATO group. Almost equal protection was observed in horsegram supplemented groups. In the ATO combination group, the protection level was further better. Also, the combination group (HG+ATO) shows the highest protection against liver damage due to obesity. Thus, showed the hepatoprotection.
Figure 8 shows the histopathologica 1 changes in Brown Fat Tissue. Adipose tissue stained with haematoxylin and eosin (Magnification =200X). Arrows and * denotes, in normal, brown fat adipocytes show rarefied cytoplasm and centrally placed small dark nuclei (arrow). White fat cells present among brown fat show normal size and shape (*). In HFD group shows brown adipose cells were smaller in size (arrow) compared to normal. White adipose cells with largë vacüólated cytoplasmic area and peripheral flat nuclei (*) was predomiriant. in ATO, marked whitening of brown fat. Brown fat cells (arrow) are few and large white fat

cells are predominant (*). In, HG + ATO, minimal whitening of brown fat observed in this group.
From the figure 8, it can be inferred that the two types of fats, they are WAT distribution greatly affects metabolic risk. BAT (good fat) important for thermogenesis and energy balance. But during high fat condition brown fat gets converted to white fat known as whitening of brown fat, "whitening" that eventually led to a functional shift from thermogenesis towards lipid storage. Brown adipose tissue (BAT) is the most effective therapeutic tooi to fight against diabetes and obesity through its capabilities to enhance its glucose expenditure. The brown adipose cells in the HFD groups were smaller in size compared to the normal diet group. White adipose cells with large vacuolated cytoplasmic area and peripheral flat nuclei was predominant (whitening of brown fat) over the brown adipose cells.
Varied levels of protection from whitening of brown fat was observed in both horsegram treated groups as well as in its combination group. However, ATO alone group showed highest level of whitening of brown fat compared to all other groups. This shows the side effect of statin intake. Thus, from our results it is observed that the combination of (HG+ATO) will prevent the whitening of brown fat thus in turn prevent obesity.
Further, other experimental results and the effect of intake of norse gram are tabulated beïow.
Table 1: Effect öf horsegram on Feed Intake:

From the Table 1, it is evident that the food intake as a behaviour is subjected to conscious contrei. The feed intake in HFD groups was significantly increased when compared to Normal diet fed rats. It should be noted that Legumes, a source of crude fibre and resistant starch delays gastric emptying and induce a sense of satiety and hencë the feed intake has been reduced in horsegram treated and its combination groups. Also, the combination of both horsegram with atorvastatin showed significant reduction in the feed intake when compared to other groups proves that it possesses a sense of satiety.
Values were expressed in Mean±SEM, n=6 NC and n=8 in all other groups, Stati'stical analysis was performed using one-way ANOVA foliowed by Tukey's multiple comparison test.
No significant inference in the water intake since there is big difference associated with the effect of horsegram on water intake.

Obesity may also lead to organ damage and accumulation of fats. Thus, in order to check its damage and degree of protectivity by legume treated groups the organ weights were measured. It is observed from the Table 3, it is clear that no treatment related significant changes in heart, kidney and brain weight were observed. However, the liver weight was reduced in the combination group compared to the HFD fed rats which probably mediated by liver damage in atorvastatin treated rats and which is reduced in the combination group considerably.
Obesity is determined by the amount of lipid stored in adipocytes and other tissues. It is observed that Horsegram individually and in combination with ATO (Horsegram+ATO) significantly resisted the increase in fat mass gain.

Values are expressed in Mean±SEM, n=6 animals/group, Statistical analysis was performed using one way ANOVA foliowed by Tukey's multiple comparison test, #, ffl and ### indicates p value< 0.05,0.01 and 0. 0.001 respecttvely Vs Normal Control; *,** and *** indicates p value ■£ 0.001 Vs HFD group.
The insulin resistance was reduced after the treatment period (28th day). HG+ATO
combination treated group showed a significant reduction in the insulin resistance and serum
glucose levels. Thereby modulates metabolic disorders such as type 2 diabetes and
cardiovascuiar disorders.

The serum enzyme levels of liver often gives an idea on the liver damage or injury. Liver ftinction tests which includes the activity of ALT, AST, ALP, y-GT and LDH were performed to confirm the degree of liver damage. Hence, horsegram with atorvastatin in HFD rats prevented the increase of biomarkers in the circulation confirming hepatoprotectivity.

From the tables, 7,8 and 9, it can be inferred that the oxidative stress plays a prodigious role in the development of metabolic disorders. It is an imbalance between production of oxidant derivatives and antioxidants defences and the protective effect of combination has been évidenced in all the tissue as évidenced by the oxidative stress markers.
As évidenced by the in vitro and in vivo studies and form the observations in the form of tables and figures, legumes are potent functional foods for management of obesity. The legumes individually as well as its combination with atorvastatin with synergistic effects they produce are heïpful to mitigate dyslipidemia and associated CVD risk factors. The combination therapy of the legumes and more specifically norse gram extends its therapeutic potential over obesity and its complications and proves to be hepatoprotective, cardioprotective and nephroprotective compared to the allopathie drug atorvastatin alone.
Ünless otherwise defïned, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs. As uséd in this specification and the appertded claims, the singular forms "a," "an,"

and "the" include plural references unless the context clearly dictates otherwise. Any referehce to "or" h'erëih is intended to encórhpass "and/or" unless otherwise sta te d.
While the preferable embodiments of the current invention have been discussed and described as appropriate, it will be obvious to a person skilied in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions are possible without departing from the scope of the present disclosure. Various alternatives to the embodiments of the present disclosure described herein may be employed while practicing the disclosure of the current invention. Further, it is intended that the following claims define the scope of the present disclosure and that methods and compositions within the scope of these claims and their equivalents be covered thereby.
Also, throughout the disclosure of the current invention, the numerical features are presented in a way of range format. The description in range format is merely for suitability and convenience and should not be construed as an inflexible limitation on the scope of any of the embodiments of the current invention. Accordingly, the description of a range mentioned is to be considered as disclósing all the possible subranges as well as individual numerical values within that range to the tenth of the unit of the lower limit unless the context clearly dictates otherwise.
For example, description of a range such as from 1 to 5 should be considered to have specifically disclosed subranges such as from 1 to 2, from 1 to 3, from 1 to 4S from 2 to 45 from 2 to 3, and from 3 to 4, as well as individual values within that range, for example, 1.1, 2, 2.3, 4, and 4.9. This applies regardless of the breadth of the range. The upper and lower liinits öf these intervénihg ranges rriay iridependently be included in the smaller ranges, and are also encompassed within the present disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the present disclosure, unless the context clearly dictates otherwise.

CLAIMS
i. A combination dosage for reducing or eliminating rises in cholesterol in the treatment of obesity and related pathologies comprising (a) at least one statin and (b) at least one fünctional food.
2. The combination dosage and treatment method of claim 1 wherein the statin is selected fröm the gróup of atorvastatin, levostatin, fluvastatin, pravastatih, rosuvastatin, or simvastatin or a pharmaceutically acceptable salt thereof.
3. The combination dosage and treatment method of claim 1 wherein the statin is atorvastatin or a pharmaceutically acceptable salt thereof.
4. The combination dosage and treatment method of claim 1 wherein the fünctional food is selected form the group of legumes.
5. The combination dosage and treatment method of claim 1 wherein the legume selected is Horsë gram.
6. The combination dosage and treatment method of claim 1 wherein the dose of Hörsë gram is in the range óf 100-400 mg/kg' of thé bódy weight and thé dóse óf atorvastatin or a pharmaceutically acceptable salt thereof is in the range of 5-15 mg/kg of the body weight.
7. The combination dosage and treatment method of claim 1 wherein the dose of Horse gram preferably 250- 350 mg/kg of the body weight and the dose of atorvastatin or a pharmaceutically acceptable salt thereof is in the range of 8-12 mg/kg of the body weight.