Capsicum compositions and uses thereof
Field
Capsicum compositions comprising of non-capsaicinoids and capsaicinoids in ratio of about 1:0.1 to 1:10 are described for the management of cardiometabolic syndrome and associated risk factors from which a subject may be suffering and may be in need of a treatment. The compositions are comprised of capsicum extract, further comprising compounds such as capsaicinoids and non-capsaicinoids in specific ratio. Non-capsaicinoids comprised in the composition are saponins and polyphenols such as flavonoids. The invention also relates to process for preparation of capsicum composition by using food grade polar and non-polar solvents. More particularly, capsicum compositions comprising biologically active constituents such as saponins, polyphenols and capsaicinoids are described herein for prevention and treatment of cardiometabolic syndrome such as hyperlipidemia, hypertension, hyperglycemia and other cardiovascular disorders. Capsicum compositions described herein improve cardiometabolic syndrome and manages associated risk factors such as body weight, body fat, lipid profile, blood glucose levels and the like, when administered to a subject in need thereof, in effective amounts. The composition also improves cardiovascular endothelial function by reducing oxidative stress and inflammatory markers. Capsicum compositions described herein can reduce cardiometabolic stress in a subject, in need thereof, by inhibiting pancreatic lipase enzyme and enhancing lipolysis. The composition is safe for consumption and can be employed for management of cardiometabolic syndrome, when administered in effective amounts.
Background
The recognition of the metabolic syndrome as a major and prevalent cause of coronary heart disease has helped to understand a key concept based on pathophysiology such as insulin resistance and connect it other related risk factors that can be diagnosed by primary care physicians. The importance of insulin resistance as a core metabolic abnormality is associated with a constellation of cardiovascular and diabetogenic risk factors/markers is also recognized and linked to abdominal obesity. In patients likely to have excess abdominal fat, the attention has to be paid to five parameters (waist circumference, triglycerides, high density lipoprotein (HDL)-cholesterol, fasting glycemia, blood pressure) in order to identify individuals with the metabolic
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syndrome, more specifically cardiometabolic syndrome. In many cases measurement of waist circumference over body mass index (BMI) is recommended when estimating the amount of abdominal fat. This recommendation to measure waist circumference or waist to hip ratio was a giant conceptual leap forward in clinical practice, as it recognized abdominal obesity as the most prevalent form of the cardiometabolic syndrome in affluent, sedentary populations.
Cardiometabolic syndrome (CMS) is thus a clustering of interrelated risk factors that promote the development of atherosclerotic vascular disease and type 2 diabetes mellitus, as a result of imbalance between energy intake and utilization. Individuals with cardiometabolic syndrome are at high risk of developing heart failure and glucose intolerance, thus affecting vital organs such as the eye, liver, kidney and nervous system. The prevalence of obesity is increasing at an alarming rate in developed and developing countries (Haslam and James, 2005). Obesity makes it more difficult for cells to respond to insulin. If the body cannot make enough insulin to override the resistance, the blood sugar level increases and diabetes can result.
Sedentary nature of work, anxiety, stress coupled with increased intake of high fat and high carbohydrate seems to be responsible for the development of cardiometabolic disorder. Free radical generation and oxidative stress is the main cause of developing associated risk factors of cardiometabolic syndrome. Hence natural antioxidant supplementations and exercise can be potential therapeutic to protect vital body organs from dreadful effects of oxidative stress and anxiety and to combat cardiometabolic syndrome.
In recent years certain chemical constituents found in natural products have attracted the interest of researchers because they show promise of being powerful antioxidants that can protect the human body from free radicals, the formation of which is associated with the normal natural metabolism of aerobic cells (Matereska 2005) The antiradical activity of compounds such as flavonoids and phenolics is principally based on the redox properties of their hydroxy groups and the structural relationships between different parts of their chemical structure. Such compounds cannot be produced by the human body and thus must be taken in mainly through the daily diet. Many such compounds having antioxidant properties are found in Pepper fruits (Capsicum annuum L.) which are important vegetables used as foods and as the spice. Peppers are a good

source of vitamins C and E as well as provitamin A. Many prior art literature references deal with evaluation of capsicum compositions in health applications.
European patent application EP2329815A1 relates to a Capsicum annuum paprika plant extract composition comprising zeaxanthin, wherein zeaxanthin is the predominant carotenoid, and wherein the Capsicum annuum paprika plant is obtained by classical plant breeding methods. The Capsicum annuum paprika plant extract composition is used for ingestion for the treatment or the prevention of human or animal diseases including cancer-related diseases, inflammatory disorders and nervous system diseases.
Another EP patent application EP1902723A1 relates to pungent principles such as capsicum, garlic and/or sabal, as such or in the form of extracts or essential oils which are used as antidiabtic, lipase inhibitors and as anorectic.
U.S. patent application US20120219621 describes a composition comprising: a) menthol or a peppermint essential oil comprising menthol, in association with one or more components having a thermogenetic activity, selected from: bl) capsaicin or a capsacinoid derived from Capsicum genus or a Capsicum extract containing capsaicin and/or capsacinoids; b2) a sulphur containing active principle derived from the Allium genus or an Allium extract containing it; b3) a catechin active principle derived from the genus Camellia, or a Camellia extract containing it, for use in the treatment and prevention of obesity, overweight, metabolic syndrome, hypercholesterolemia, hypertriglyceridemia, diabetes or hypertension. This application deals with synergistic effects of menthol or essential oil in association with either capsicum extract or sulphur or catechin for said health benefits.
Korean application KR1020130103101 deals with a composition for body fat lipolysis using alginate double-layer nanoemulsions containing oleoresin capsicum to resolve problems such as overweight and obesity. A method for manufacturing the alginate double-layer nanoemulsion composition comprises the steps of: mixing oleoresin capsicum and Tween 80 in a ratio of 1:3 to prepare a mixture solution; and mixing the mixture solution and an alginic acid solution to prepare double-layer nanoemulsions.
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U.S. patent 7892582 deals with a composition for oral administration containing capsaicinoids associated with a formulation base acceptable for oral administration, the formulation base comprising an oil and an additive solid or pasty at room temperature. Further it discloses a method for stimulating thermogenesis, treating obesity, digestive problems, or for reducing the appearance of cellulite comprising administering the said composition.
Korean patent application KR1020080048426 describes an anti-obesity composition containing fresh red pepper (Capsicum annuum L.) extract as an active ingredient. The composition contains 20 to 100% by weight of fresh red pepper extract, 20 to 80% by weight of a corn silk extract, 0.5 to 10% by weight of Pueraria root extract and 0.5 to 10% by weight of garlic extract.
PCT patent application WO2015041977 provides a composition comprising at least two compounds capable of enhancing thermogenesis or identified as capable of enhancing thermogenesis, wherein the composition comprises one non-stimulant and at least one stimulant. The non-stimulant thermogenic agent is an agent selected from the group consisting of: a) fucoxanthin; b) branched-chain amino acid; c) decaffeinated tea, decaffeinated tea extract, or isolated component or components from decaffeinated tea; and d) capsaicin extract or isolated component or components from capsaicin extracts.
PCT application WO2010128788 relates to anti-obesity external dermal agent compositions containing capsaicin or capsaicin-like compounds as active ingredients. The anti-obesity external dermal agent compositions containing capsaicin or capsaicin-like compounds according to the present invention stimulate the metabolism of the body to decompose the fat cells of subcutaneous fat, and inhibit body fat accumulation to reduce hypertrophy of fatty tissues, thus eventually reducing weight and efficiently preventing and improving obesity.
U.S. patent application US20120321730 provides an industrial process for isolation and purification of capsanthin rich carotenoid mixture from paprika oleoresin extracted from Capsicum annuum. Further it discloses a method of using Capsicum annuum extract for cosmetic application, as a nutraceutical, as an antioxidant or as a health supplement said method comprising act of contacting the extract or the composition with a subject in need thereof.
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U.S. patent 8663714 relates to a dietary supplement for treating obesity in a human in need thereof consisting essentially of therapeutically effective amounts of alginate, Capsicum frutescens extract, Mentha piperita extract and glucomacropeptide. As per this invention, extracts of chilli (Capsicum species) and mint (Mentha species) can be encapsulated into beads of alginate gel.
PCT application WO2004047717 discloses a topical application of Curcumin (from Curcuma longa, Turmeric) alone or in combination with Capsaicin (Capsicum oleoresin) for treating peripheral neuropathies, including diabetic neuropathy associated with chronic type I or Type II diabetes mellitus.
Chaiyasit et al (J Med Assoc Thai. 2009 Jan; 92 (1): 108-13) relates to the effect of capsicum on plasma glucose level and to correlate its action with the pharmacokinetic properties of capsaicin in capsicum. The results showed that plasma glucose levels in volunteers who received capsicum were significantly lower than those in the placebo group.
Tolan et al (Phytother Res. 2001 Aug; 15 (5):391-4) describes the effect of capsaicin from Capsicum frutescens on blood glucose, plasma insulin levels and insulin binding in dog models. This study was designed to identify any hypoglycaemic principle(s) and to determine the mechanism of action. It was observed that capsaicin is responsible for the hypoglycaemic episodes seen in the dogs and that it also causes an increase in insulin secretion which leads to a reduction of insulin binding on the insulin receptors.
Bloomer et al (Lipids Health Dis. 2010 Jul 15; 9:72) describes effect of oral intake of
capsaicinoid beadlets on catecholamine secretion and blood markers of lipolysis in healthy
adults. Blood epinephrine (EPI), norepinephrine (NE), free fatty acids (FFA) and glycerol
concentrations were compared in response to a capsaicinoid supplement or placebo in healthy
adults before and after acute exercise. Ingestion of low dose (2 mg) Capsicum composition was
associated with an increase in blood FFA and glycerol at selected times post ingestion, as
compared to placebo. However, the composition had no differing effect on EPI or NE compared
to placebo.
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Summary
Even though the literature above discusses the effect of capsicum in thermogenesis, body fat reduction, and neuropathy as well as for reducing body weight, there is no disclosure on chemical constituents of capsicum composition which exert these and related biological activity. Even though there are some descriptions on commonly found chemical constituents such as capsaicinoids in literature and their possible effect in weight management, there are no reports till date, which focus on characterization of capsicum extracts for identification of specific capsaicinoids and non-capsaicinoid constituents such as flavonoids and saponins and defining their specific ratio, which exhibit beneficial biological effects in the body systems, such as cardiovascular system.
Applicant has carried out rigorous experimentation and optimization for preparation and characterization of capsicum extract compositions, as described herein. The applicant has also characterized the extract composition for identification of chemical constituents by employing sophisticated analytical techniques. Capsicum extract compositions are further evaluated for exploring effect in preventing and treating cardiometabolic syndrome by effectively reducing multiple risk factors and co-relating the chemical constituents identified to this biological activity, which is not reported anywhere in the prior art till date.
Capsicum compositions of the invention are comprised of capsicum extract, further comprising biologically active chemical constituents such as non-capsaicinoids and capsaicinoids in specific ratio, non-casaicinoids are further comprised of saponins and polyphenols such as flavonoids. Capsicum composition is characterized by different analytical method to identify capsaicinoids, polyphenols and saponins. Major capsaicinoids such as capsaicin and dihydrocapsaicin and other capsaicinoids such as nor dihydrocapsaicin, decanyl vallinamide, dihydrocapsaicin, homocapsaicin are identified and quantified along with non-capsaicinoid compounds. Non-capsaicinoid compounds such as polyphenols and saponins are identified from capsicum composition. Polyphenols may be comprised of flavonoids and cinnamic acid derivatives in the composition described herein. Capsaicinoids such as capsaicin and dihydrocapsaicin are called as major capsaicinoids which are present in range of about 25 to 35 % w/w in the composition.
Other capsaicinoids are present in amount of about 0.5 to 1.5 % w/w, in said compositions. The
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compositions as described herein also contain total polyphenols comprising flavonoids and cinnamic acid derivatives, in the range of about 10 to 20% w/w. The compositions are also analyzed for saponin content and found to be in the range of 0.1 to 5% w/w. Capsicum compositions are safe for human consumption and are comprised of biologically active chemical constituents non-capsaicinoids and capsaicinoids in the ratio of about 1:0.1 to 1:10, such composition with specific ratio of active constituents, which manages cardiometabolic syndrome is reported for the first time.
Capsicum compositions described herein when administered in effective amount to a subject in need thereof, helps to improve cardiometabolic syndrome and manages associated risk factors such as body weight, body fats, lipid profile, blood glucose and the like. The composition also improves cardiovascular endothelial function by reducing oxidative stress and inflammatory markers. Capsicum compositions described herein can reduce cardiometabolic stress in a subject, in need thereof, by inhibiting pancreatic lipase enzyme and enhancing lipolysis. The composition is prepared by human intervention using food grade solvents, it is safe for human consumption and can be employed for management of cardiometabolic syndrome, when administered in effective amounts.
Objectives of the present invention
The main objective of the present invention is to provide capsicum compositions having specific ratio of saponins, polyphenols and capsaicinoids, which when administered to a subject in need thereof, are useful for management of cardiometabolic syndrome and associated risk factors.
Another object of the present invention is to provide capsicum composition comprising capsicum extract, further comprising chemical constituents like capsaicinoids and non-capsaicinoids. The capsaicinoids are selected from the group of, but not limited to compounds such as capsaicin, dihydrocapsaicin, nordihydrocapsaicin, homocapsaicin, dihydrohomocapsaicin, 6",7"-dihydro-5',5'"-dicapsaicin, 5,5'-dicapsaicin, decaylvanillamide; while non-capsaicinoid compounds are saponins and polyphenols such as flavonoids, which are biologically active compounds, identified for their effect in management of cardiometabolic syndrome, when administered in effective amounts.
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As per one more important object of the invention, capsicum extract composition is prepared from various varieties of paprika selected from the group of, but not limited to, varieties such as Teja, Namdhari and so on. More particularly, the composition may be comprised of the capsicum extract obtained from varieties of Capsicum annum such as Teja and Namdhari.
One more objective of the present invention is to provide an industrially viable process for preparation of capsicum composition by employing non-polar, semi-polar, polar solvents or combinations thereof in suitable ratios. Still one more objective of the present invention is to provide capsicum composition prepared using combination of polar and non-polar solvents in suitable ratio for extraction cycle, followed by purification of extract by using polar solvents to get the extract. The composition is safe for human consumption and is comprised of biologically active chemical constituents such capsaicinoids and non-capsaicinoids, which are useful for management of cardiometabolic syndrome in subjects, when administered in effective amounts.
Yet one more objective of the present invention is to provide capsicum composition comprised of chemical constituents, which are characterized by various analytical methods.
Yet another object of the invention is to provide capsicum composition which is prepared by using capsicum extract either alone or in combination with other nutrients such as capsanthin, capsanthin-ester, betacryptoxanthin, betacarotene, capsorubin, zeaxanthin, antheraxanthin, and/or at least one more pharmaceutically or nutraceutically acceptable excipients and formulating it in suitable delivery form for oral or non-oral route of administration.
Yet one more important objective of the invention is to provide methods for use of capsicum compositions comprising biologically active constituents such as capsaicinoids and non-capsaicinoids and evaluate its effects on management of cardiometabolic syndrome and the associated risk factors, in a subject in need thereof.
Yet one more objective of the invention is to provide methods for use of capsicum compositions
comprising biologically active constituents in managing, treating or improving conditions such
as hypertension, hyperlipidemia, hyperglycemia, insulin resistance, obesity, waist to hip ratio,
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and the like, associated with cardiometabolic syndrome by administering effective amounts of capsicum compositions, to a subject in need thereof.
Detailed Description of the Figures
Figure 1 depicts effect of capsicum composition on pancreatic lipase enzyme Figure 2 depicts effect of capsicum composition on gene expression Figure 3 depicts effect of capsicum compositions on gp91 phox and NFkB Figure 4 depicts effect of capsicum compositions on Nrf2 and SIRT 1 Figure 5 depicts effect of capsicum compositions on eNOS and p22 phox Figure 6 depicts effect of capsicum compositions on calorie intake in females Figure 7 depicts effect of capsicum composition on Diastolic Blood pressure Figure 8 depicts effect of capsicum composition on hip circumference Figure 9 depicts effect of capsicum composition on Waist to Hip ratio
Detailed Description
Compositions and methods herein are directed to preparing capsicum compositions comprising
specific biologically active chemical constituents, which are effective in treating and/or
managing cardiometabolic syndrome and their associated risk factor(s). The compositions are
comprised of capsaicinoids and non capsaicinoids. Further capsaicinoids are selected from group
of, but not limited to, the compounds such as capsaicin, dihydrocapsaicin, nordihydrocapsaicin,
homocapsaicin, dihydrohomocapsaicin, 6",7"-dihydro-5',5"'-dicapsaicin, 5,5'-dicapsaicin,
decanylvallinamide and non-capsaicinoid compounds like polyphenols and saponins in specific
ratio. Polyphenols present in capsicum composition may be in the form of flavonoids and
cinnamic acid derivatives. The compositions described herein are comprised of biologically
active chemical constituents which are useful for managing cardiometabolic syndrome and
associated risk conditions such as body weight, lipid profile, insulin resistance and blood glucose
and the like, when administered in effective amounts to the subjects in need thereof. The
composition exhibits reduction in oxidative stress and inflammatory markers, enhances lipolysis
and inhibits pancreatic lipase when administered to a subject, in effective amounts.
Capsicum compositions herein are obtained by human intervention, using conventional industrial
equipments and are safe for human administration and thus useful for nutraceutical purposes.
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Cardiometabolic syndrome, also known as syndrome X, increases the risk of developing cardiovascular disease, particularly atherosclerosis, heart failure, dyslipidemia, diabetes, and associated risk factors, which may be caused mainly due to imbalance of calorie intake and energy utilization. One of the most important causes for this is wrong dietary habits.
The terminology 'subject' is commonly used in the specification to refer to an individual or mammal under test, being treated with compositions herein. The terminology "subject in need thereof can include specific individuals or mammals, may be are habituated to a diet rich in high fat and refined carbohydrates, thus lacking in fibers. Such subjects are at high risk of developing and/or may be suffering from cardiometabolic syndrome, because of developing abdominal obesity. These subjects also exhibits disturbed body parameters such as blood glucose, body fats, lipid profile, increased body weight and are there prone to cardiovascular diseases.
The terminology "capsaicinoids" as used herein refers to the chemical compounds identified in capsicum extract prepared from paprika variety, which belong to classes selected from, but not limited to, capsaicinoids, dicapsaicinoids, capsaicin, nor dihydrocapsaicin, decanylvallinamide, dihydrocapsaicin, homocapsaicin their derivatives, salts and the like.
The term "non-capsaicinoids" as used herein refers to the chemical compounds identified from capsicum extract, which belong to classes other than capsaicinoids and these compounds can be saponins, polyohenols such as cinnamic acid derivatives and flavonids, and the like, which in combination with capsaicinoids exert biological activity and are effective in treatment and management of cardiometabolic syndrome, when administered to a subject, in need of treatment thereof.
Abdominal obesity drives the progression of multiple risk factors directly, through the secretion of excess free fatty acids and inflammatory adipokines, and decreased secretion of adiponectin. Excess abdominal obesity is accompanied by elevated levels of C-reactive protein (CRP) and free fatty acids (FFAs), as well as decreased levels of adiponectin. Elevated CRP is an indicator of inflammation. Abdominal obesity may be associated with the inflammation cascade, with adipose tissue expressing a number of inflammatory cytokines. Inflammation is now believed to play a role in the development of atherosclerosis and type 2 diabetes. Elevated levels of CRP are considered to be predictive of cardiovascular disease and insulin resistance.
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Low-grade chronic inflammation is characterized by a two- to threefold increase in the systemic concentrations of cytokines such as TNF-a, IL-6, and/or CRP. TNF's primary role is to regulate the immune cells and induce inflammation. TNFa -induced reductions in insulin sensitivity in adipocytes are partly responsible for the increased free fatty acid production and hypertriglyceridaemia characteristic of abdominal obesity. Leptin responds specifically to adipose-derived inflammatory cytokines. Hyperglycemia induces IL-6 production from endothelial cells and macrophages. Meals high in saturated fat, as well as meals high in calories have been associated with increases in inflammatory markers.
As per one embodiment, capsicum compositions and methods of use thereof, when administered to a subject in need thereof, in effective amounts, treat and/or manage risk factors associated with cardiometabolic syndrome.
As per one more important embodiment, capsicum composition is mainly comprised of capsicum extract prepared from various varieties of paprika, which is further comprised of chemical constituents such as non-capsaicinoids in combination with capsaicinoids in a ratio of about 1:0.1 to 1:10. Thus capsicum compositions are comprised of capsicum extract rich in biologically active constituents such as capsaicionoids, saponins and flavonoids, which is formulated into suitable dosage form and/or delivery system, by combining with other nutrients along with other pharmaceutically or nutraceutically acceptable excipients.
According to one embodiment, capsicum compositions are comprised of capsicum extract and one or more nutrients such as carotenoids, omega-3-fatty acids, vitamins, minerals, which may improve biological effect of capsicum extract synergistically.
According to still one more embodiment, capsicum compositions are also comprised of capsicum extract and one or more nutrients, formulated along with at least one more pharmaceutically or nutraceutically acceptable excipient selected from the group of, but not limited to diluent, antioxidant, binder, film former, carrier, buffer, vehicle, stabilizer, surfactant, glidant, filler and the like or the combination thereof.
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According to one embodiment, capsicum extract is prepared from different varieties of raw paprika such as Teja and Namdhari by employing organic and/or inorganic solvents selected from the group of, but not limited to polar, semi-polar and non-polar solvents or the combination thereof.
As per one more embodiment, capsicum extract is prepared by extracting deseeded and pulverized raw paprika with combination of polar and non-polar solvents. Polar solvents used in the process may be selected from the group of, but not limited to, alcohol, water, acetone, ethyl acetate, acetonitrile, dimethylformamide, tetrahydrofuran and the like, either alone or in combination thereof.
As per still one more embodiment, capsicum extract is prepared by extracting paprika using a non-polar solvent selected from the group of, but not limited to, pentane, hexane, cyclohexane, diethyl ether, and the like, either alone or in the combination thereof.
In one preferred embodiment, capsicum extract is prepared by employing combination of polar and non-polar solvent in a ratio of 1:0.5 to 1:10 by weight.
In one more embodiment, the ratio of paprika raw material to solvent employed is about 1:0.5 to 1:20 by weight in each extraction cycle. Sufficient numbers of extractions are carried out followed by percolation.
As per one important embodiment of the invention, capsicum composition comprising extract is prepared by extracting paprika raw material employing combination of polar and non-polar solvents such as acetone and hexane, in suitable ratio, in each extraction cycle and extracted mass from all such extractions are combined, distilled and concentrated. Crude extract thus obtained is washed with polar solvents such as alcohol and water to get purified capsicum extract, which can be either used alone or in the form of dosage form, prepared using at least one excipient and carrier, for convenient administration to the subject in need thereof.
According to one embodiment, capsicum extract is characterized by analytical techniques such as TLC (thin layer chromatography), MS (Mass spectroscopy), gravimetric,
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UV spectrophotometry and other suitable techniques for identification of capsaicinoids and non-capsaicinoid compounds.
According to one embodiment, capsicum compositions comprising extract, can be formulated as a pharmaceutical or nutraceutical delivery systems, in the form of dietary supplement, dosage form or in suitable vehicle, convenient for administration. The compositions can be administered in the form of powders, granules, sachets, beadlets, tablets, capsules, caplets, suspensions, emulsions, solutions, energy bar, beverages, functional foods and the like.
Capsicum composition of the invention may be administered by oral route or the other suitable routes other than oral, in combination with antioxidant or other nutrients, using suitable vehicle for administration.
As per one embodiment, the compositions described herein are comprised of biologically active chemical constituents which can help to improve risk factors associated with cardiometabolic syndrome, such as body weight, lipid profile, body glucose, and/or insulin resistance, when administered to a subject.
As per one important embodiment, the methods described herein are directed to identifying a subject in need thereof, for treatment, prevention or improvement of a condition or risk factor associated with cardiometabolic syndrome, and administering capsicum composition, comprising biologically active constituents, in effective amounts, to the subject in need thereof.
Capsicum composition is characterized by different analytical method to identify capsaicinoids, polyphenols and saponins. Major capsaicinoids such as capsaicin and dihydrocapsaicin and other capsaicinoids such as nor dihydrocapsaicin, decanyl vallinamide, dihydrocapsaicin, homocapsaicin are identified and quantified along with non-capsaicinoid compounds. Non-capsaicinoid compounds such as polyphenols and saponins are identified from capsicum composition. Polyphenols may be comprised of flavonoids and cinnamic acid derivatives in the composition described herein. Capsaicinoids such as capsaicin and dihydrocapsaicin are called as major capsaicinoids which are present in range of about 25 to 35 % w/w in the composition. Other capsaicinoids are present in amount of about 0.5 to 1.5 % w/w, in said compositions. The
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compositions as described herein also contain total polyphenols comprising flavonoids and cinnamic acid derivatives, in the range of about 10 to 20% w/w. The compositions are also analyzed for saponin content and found to be in the range of 0.1 to 5% w/w. Thus capsicum compositions are comprised of non-capsaicinoids and capsaicinoids in the ratio of about 1:0.1 to 1:10, such composition with specific ratio of biologically active chemical constituents is useful for management of cardiometabolic syndrome.
According to one important embodiment, capsicum compositions are comprised of 0.01% to 75% bioactive compounds by weight in capsicum extract.
According to one more embodiment of the invention, capsicum compositions comprising capsaicinoids and non-capsaicinoid compounds are administered in effective amounts, such as, 0.5 mg to 10 mg/kg body weight to a subject in need thereof.
According to one embodiment, capsicum compositions comprising about 0.01 to 75% biologically active compounds, are administered to a subject, in need of treatment of cardiometabolic syndrome, in the form of dietary supplement either alone or optionally in combination with ingredients selected from the group of, but not limited to lipid lowering, blood sugar lowering, blood pressure lowering ingredients and the like or the combinations thereof.
As per one embodiment, capsicum compositions and methods herein can treat hyperlipidemia, in a subject in need thereof, when administered in effective amounts, by lowering total cholesterol, low density lipoproteins and/or triglycerides.
According to one more embodiment, compositions and methods herein are directed to reducing free fatty acid levels, and/or visceral fat, along with body weight, when administered in effective amounts to the subjects in need thereof.
According to still one more embodiment, capsicum compositions described herein are also effective in inhibiting fat absorption by inhibition of pancreatic lipase and enhancement of lipolysis and thermogenesis, when administered in effective amounts, to a subject in need thereof.
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According to one embodiment, capsicum compositions described herein are effective to improve post-prandial hypoglycemia and insulin sensitivity in a subject, in need thereof by the reduction in body glucose levels and/or the reduction in insulin resistance and protect vital body organs, when administered in effective amounts.
According to one embodiment, capsicum compositions and methods herein are also used to treat and/or evaluate their effect on expression of inflammatory markers and/or oxidative stress markers, when administered to a subject in need thereof. The compositions are also evaluated in cell line studies for checking effect on enzymes or cardiometabolic markers. The evaluation is also carried out in human volunteers for their effect on various enzymes and effect on body fats, blood pressure, waist to hip ratio, appetite and other cardiomatabolic risk factors to understand effect for management of cardiometabolic syndrome.
For evaluation purposes, the compositions may be administered in the form of semi-solid concentrates or the compositions suspended in suitable oil vehicle or the formulations such as beadlets or capsules. The formulation may be also mentioned as Capsimax or CAPs to denote capsicum compositions in the specification.
In one embodiment, compositions and methods herein are directed to evaluating the effect(s) of capsicum composition on the improvement of cardiometabolic health by administering to a subject in need thereof, an effective amount of a composition comprising capsicum alone or in combination with other nutrients.
In one embodiment, capsicum compositions herein and methods of use thereof are directed to the improvement of cardiometabolic health by management of a healthy lipid profile, reduction in body fat, visceral fat, and free fatty acid levels in the body.
In one embodiment, capsicum compositions herein and methods of use thereof are directed to
administering the composition, to a subject in need thereof, for the treatment and/or management
of cardiometabolic syndrome, to improve lipid profile, to reduce body weight, and/or to reduce
oxidative stress markers in the vital body organs.
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In one embodiment, capsicum compositions and methods of use thereof are directed to reducing risk factors associated with cardiometabolic syndrome, such as for example obesity, hyperglycemia, hypertension, and/or hyperlipidemia, and the like.
In some embodiments, compositions herein include a capsicum extract administered either alone or in the form of composition with other nutrient and/or food grade excipients. Other nutrients in capsicum compositions can be capsanthin, capsanthin-ester, betacryptoxanthin, betacarotene, capsorubin, zeaxanthin, antheraxanthin. Compositions are comprised of extracts, further comprising of biologically active constituents such as capsaicinoids and non-capsaicinoids. The compositions may be administered for a period of about 7 days to about 12 months. Preferred time of administration is with meals.
Capsicum compositions, as described herein are safe for human consumption and are prepared using food grade solvents. These are comprised of biologically active chemical constituents such as non-capsaicinoids and capsaicinoids in a ratio of about 1:0.1 to 1: 10 and are useful for management of cardiovascular syndrome and associated risk factors, when administered to the subjects in need thereof in effective amounts. The compositions are analyzed for identification of chemical constituents and also evaluated for biological activity through in-vitro and in-vivo evaluations.
While the compositions and methods herein have been described in terms of specific illustrative embodiments, any modifications and equivalents that would be apparent to those skilled in the art are intended to be included within the scope of the compositions and methods herein. The details of the compositions and methods herein, its objects, and advantages are explained hereunder in greater detail in relation to non-limiting exemplary illustrations.
Examples
Example 1: Preparation of Capsicum composition
Capsicum composition is prepared by extracting dried pods of capsicum annum with combination of acetone and hexane in a ratio of 1:0.5 to 1:10 by weight and it is subjected to
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filtration. The ratio of raw material to solvent employed is about 1:0.5 to 1:20 by weight in each extraction cycle. Sufficient numbers of extractions are carried out at suitable process conditions followed by percolation. Extracted mass from all such extractions are combined, distilled and concentrated. Crude extract thus obtained is washed with suitable polar solvents to get purified capsicum composition. This method was used for preparation of all three batches CC1 to CC3 as shown in Table no. 1.
Example 2a: Determination of capsicum composition for capsaicinoids
Capsicum composition is analyzed by HPLC method using Thermo CI8 column at 281 nm detector wavelength. Methanol was used as diluent and mobile phase was prepared using 0.1% Ortho Phosphoric Acid in Water and acetonitrile in a ratio of 600:400. Capsaicin Solution was used as a standard, which was prepared by dissolving reference standard in Methanol. About 25 mg of capsicum composition was accurately weighed and diluted with methanol to get test solution.
Equal volumes of standard Capsaicin solution, Standard dihydrocapsaicin solution and test Solution were separately injected into the chromatograph and the area of the responses for all of the peaks was measured to calculate the percentage of capsaicin, dihydrocapsaicin and other capsaicinoids.
Example 2b: Determination of capsicum composition for non- capsaicinoids (polyphenols)
Polyphenol content was determined as total polyphenol content (as Gallic acid equivalent) by UV spectrophotometry technique by Folin-Ciocalteau's phenol reagent method. Capsicum composition is dispersed in methanol in desired concentration and 1.0 ml of each sample is taken into test tube and mixed with 5 ml of a 10 fold dilute Folin-Ciocalteu reagent and 4 ml of 7.5% sodium carbonate. The tubes are covered with parafilm and allowed to stand for 50 minutes at room temperature. Then the absorbance was read at 765nm spectrometrically against Gallic acid as a standard.
Flavonoids were identified by LCMS method from batch CC3 and the compound was identified as Apigenin 6-C-p-D-glucopyranoside- 8-C-a-L-arabinopyranoside.
Example 2c: Determination of capsicum composition for non- capsaicinoids (saponins)
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Saponins were quantified using gravimetric method. Capsicum composition was extracted with methanol by refluxing for an hour and the resulting residue was extracted two more times by using methanol. Methanolic extract was combined and precipitated with acetone. The precipitate was filtered and weighed.
Table 1: Chemical composition (% w/w) of capsicum composition

Compos ition Capsaic in Nor
dihydro capsaici n Decany lvallina mide Dihydro capsaici n Homo capsai cin Total capsaicin oids as capsaicin Total
Polypheno
l(As
Gallic
acid) Total
Sapon
ins
CC1 20.65 0.83 0.0004 10.98 0.0002 32.46 13.36 1.09
CC2 19.09 1.62 0.0099 13.61 0.0000 34.33 15.96 0.89
CC3 21.00 1.00 0.0042 10.95 0.0004 32.95 13.90 0.95
(cc-capsaicin composition)
The Capsicum composition is comprised of:
1) Major Capsaicinoids (Capsaicin & Dihydrocapsaicin) = between 25 - 35%w/w.
2) Other Capsaicinoids (Nor dihydrocapsaicin + Decanylvallinamide + Homocapsaicin) = between 0.5-1.5% w/w.
3) Total Polyphenols (As gallic acids) = between 10 to 20%w/w.
4) Total saponins= between 0.1 to 5%
Ratio of non-capsaicinoids: Capsaicinoids = 1:01 to 1:10
Example 3: Evaluation of capsicum composition for activity on pancreatic lipase enzyme and on lipolysis
Pancreatic lipase activity using fluorescence assay: Pancreatic lipase activity was measured using 4-methylumbelliferyl oleate (MU Oleate) as a substrate via a fluorescence kinetic assay. Orlistat was used as positive control.
Lipolysis assessment using a cell based assay: Preadipocytes 3T3L1 cells were used and differentiated, then treated with capsicum composition (lOOug/ml) at different time points such
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as lh, 2h, 4h and 24h. Lipolysis was assessed using glycerol release as a biomarker. Isoproterenol was used as positive control.
Lipolysis was assessed using glycerol release as a biomarker.
Observations: Capsicum composition displayed a strong potent inhibitory effect of pancreatic lipase (PL) compared to Orlistat. IC50 for capsicum composition (CC1) was 5.4 (|^g/ml) compared to Orlistat (0.53 (J-g/ml).
Table 2: Effect of capsicum composition on lipolysis

Details (composition and time) Ratio treated /non treated
Capsicum composition lh 0.80
Capsicum composition 2h 1.00
Capsicum composition 4h 1.31
Capsicum composition 24h 1.59
Isoproterenol (positive control) 1.60
Capsicum composition enhanced the lipolysis after 24h treatment. The ratio of released glycerol/protein content was 1.59, compared to positive control: isoproterenol (ratio: 1.60).
Gene Expression Study: 3T3L1 pre-adipocytes were treated overnight with capsicum composition CC1. RNA was isolated and cDNA prepared. Real time PCR was performed on a selected list of genes using standard protocols.
Observation: Capsicum compositions exhibited reduced fat accumulation by down-regulating PPARy and C/EBPa in 3T3L1 adipocytes.
These observations suggest that capsicum composition has inhibitory effects on pancreatic lipase, decreases fat accumulation by adipocyte differentiation inhibition and stimulates lipolysis on adipocytes. Therefore, capsicum composition could be developed as a functional food in
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helping the treatment of postprandial lipidemia and/or prevention of weight gain and weight management.
Thus it was found that capsicum compositions inhibit pancreatic lipase and also decrease mRNA levels of adipogenic genes such as peroxisome proliferator-activated receptor-gamma (PPAR-gamma) and CCAAT enhancer-binding protein-alpha (C/EBP-alpha) when studied in in-vitro models. (Capsimax represents capsicum composition of the invention)
Example 4: Effect of capsicum composition on Cardiovascular markers
Human Umbilical Vein Endothelial Cells (HUVEC) were purchased from Clonetics (Cambrex, Inc., Walkersville, MD, USA), defrosted and cultured in suitable growth medium. The cells were passaged three times to provide a pool of cells. Experiments were performed in six-well plates (Corningjnc, Corning, NY, USA) when the cells were 80% confluent, at which stage the endothelial basal medium 2 growth media (2 % fetal calf serum, with growth factors) was replaced with endothelial basal medium 2 control media (0-8 % fetal calf serum, no growth factors) and incubated for 24 h (37C, 5% C02). Treatments were prepared in endothelial basal medium 2 containing 0.8 % fetal calf serum and antibiotics but no other supplements. Capsicum compositions were used as test samples to treat the cells. Eighteen hours after treatment RNA was extracted. Real-time PCR (polymerase chain reaction) was performed on a select list of genes using standard protocols.
Observation: capsicum composition decreased VCAM1, ICAM1 and increased eNOS, which is an indicative of improving cardiovascular health.
It was also observed that capsicum compositions can prevent low-fat/high-carbohydrate diet and high-fat diet-induced obesity by regulating the expression of genes for metabolism.
Experimental diets supplemented with capsicum compositions lowered the levels of mRNA levels of gp91phox (NOX2), NFkB, p22phox Protein (CYBA) and However, the mRNA levels of SIRT which is an important regulator of energy metabolism, Nrf2 an emerging regulator of cellular resistance to oxidant, eNOS (an enzyme involved in the synthesis of a vasoprotective molecule: nitric oxide) were significantly increased.
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Capsicum composition treated groups had significant decrease in TBARS in serum and liver tissue and an increase in serum TAC were observed.
These results suggest that capsicum composition effectively reduces TBARS and increased TAC. These effects might be at least partially mediated via regulation of the gene proteins involved in energy metabolism, oxidative stress and vasoprotection.
Example 5: Effect of capsicum composition on Body indices, glucose management and blood pressure
In a double blind, randomized, placebo controlled design seventy-seven (29.6 ± 11.3 yrs, 171.2 ± 9.8 cm, 80.9 ± 18.9 kg, 27.4 ± 5.4 kg/m2) apparently healthy males and females were randomly assigned by fat mass to ingest either 2 mg/kg body weight capsicum composition or 4 mg/kg body weight composition or placebo (maltodextrin, PLA) for 12 weeks. Subjects were requested not to make any changes to their current physical activity, provided instructions to restrict spicy foods and maintain current caloric intake. Subjects were instructed to take the pills with 8 ounces of water after breakfast. At baseline (Tl), 6 weeks (T2) and 12 weeks (T3), waist and hip circumferences (WC, HC, and WHR), weight, appetite levels (Council on Nutrition appetite questionnaire -CNAQ) and adverse events questionnaires were administered. At baseline (Tl) and 12 weeks (T3) complete blood count and complete metabolic panels were assessed. Statistical analyses utilized a two-way ANOVA (group x time) with repeated measures for all dependent variables {p< 0.05).
Waist: hip ratio measurements showed a main effect for time (p = 0.034) with post-hoc tests revealing a significant (p = 0.024) decrease in results from baseline to 6 weeks interval. Appetite questionnaire responses indicated a significant main effect for time from Tl to T2 (p = 0.004) and Tl to T3 (p = 0.001) suggesting supplementation of CAP reduced appetite. Post-hoc analysis showed significant decreases in the following: PLA (Tl to T2, p = 0.05), composition dose 2 mg/kg body weight (Tl to T3, p = 0.018), and composition dose 4 mg/kg body weight (Tl to T2, p = 0.029; Tl to T3, p = 0.04). Dietary recall analysis of total caloric intake showed a significant main effect for time (p = 0.019) in females at the 4 mg/kg body weight capsicum composition dose at T3 (12 weeks).
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Capsicum composition dose decreased diastolic blood pressure and changes in glucose metabolism observed. No significant changes were observed in clinical blood safety markers at both doses of CAP. These findings show evidence to support that the dietary supplementation of CAP has beneficial effects on anthropometric parameters (waist and hip circumferences) and appetite suppression. The results also exhibited significant reduction in total caloric intake over 12 weeks.
Even relatively small reductions in blood pressure (systolic blood pressure 10-12 mmHg, diastolic blood pressure 5-6 mmHg) substantially reduce cardiovascular risk in the subjects in need thereof. Thus capsicum composition has beneficial effect in management of cardiometabolic syndrome, when administered in effective amount to the subjects.
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Claims:
1. A capsicum composition, comprised of biologically active chemical constituents such as non-capsaicinoids and capsaicinoids in a ratio of about 1:0.1 to 1:10, which is safe for human consumption and manages cardiometabolic syndrome and associated risk factors, when administered to a subject in need thereof in effective amounts.
2. Capsicum composition of claim 1, which is comprised of non-capsaicinoids such as saponins and polyphenols in range of about 10 to 25% w/w of total composition.
3. Capsicum composition of claim 2, which is comprised of polyphenols such as flavonoids and cinnamic acid derivatives in range of about 10 to 20% w/w of total composition.
4. Capsicum composition of claim 1, which is comprised of capsaicinoids such as capsaicin, dihydrocapsaicin, homocapsaicin, nor dihydrocapsaicin and decanylvallinamide in range of about 25 to 40% w/w of capsicum composition.
5. Capsicum composition of claim 1, which is safe for human consumption and manages cardiometabolic syndrome and associated risk factors such as blood glucose, lipid profile, body fat, body weight and the like, when administered in effective amounts to the subjects in need thereof.
6. Capsicum composition of claim 5, which is safe for human consumption and manages cardiometabolic syndrome through reduction .in oxidative stress and inflammatory markers, when administered in effective amount to the subject in need thereof.
7. Capsicum composition of claim 5, which manages cardiometabolic syndrome through enhancement of lipolysis and inhibition of pancreatic lipase enzyme, when administered to the subject in need thereof in effective amounts.

8. Capsicum composition of claim 1, which is administered in effective dose of 0.5 mg to 10 mg/kg body weight, for management of cardiometabolic syndrome and associated risk factors such as blood glucose, body fat, body weight, lipid profile and the like, to the subjects in need thereof.
9. A process for preparation of capsicum composition, wherein
i. Capsicum pods are extracted more than one time by using combination of polar and
non-polar solvents in suitable ratio, ii. the extracted mass is distilled and concentrated. iii. Concentrated mass is washed with polar solvents to get purified capsicum extract
10. The process of claim 9, wherein polar and non-polar solvents used for extraction are
acetone and hexane, which are used in a ratio of about 1:0.5 to 1:10.