Synergistic herbal compositions for the treatment of obesity and overweight - Patent Application 20070122997

Abstract

To provide synergistic herbal compositions for treatment of obesity and overweight.SOLUTION: The invention discloses synergistic herbal composition comprising combination of first ingredient selected from extracts, fractions, phytochemicals and mixtures thereof derived from Theobroma cacao and a second ingredient selected from extracts, fractions, phytochemical and mixtures thereof derived from Citrus aurantifolia for obtaining at least one health benefit selected from preventing, controlling or treating obesity and / or overweight; improving lean body mass, improving the browning of white adipose tissue / improving formation of brown adipose tissue, increasing basal metabolic rate / resting energy expenditure, increasing thermogenesis, improving thyroid function, maintaining healthy body weight, increasing satiety, supporting weight loss, improving fat loss and maintaining a slim body.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention provides a method for preventing, controlling, or treating obesity and / or overweight and improving lean body mass. Improves the browning of white adipose tissue (WAT) and promotes the formation of brown adipose tissue (BAT) Improves metabolism, increases basal metabolic rate (BMR), stabilizes energy expenditure, and increases thermogenesis Improves thyroid function, maintains a healthy weight, increases satiety, and aids in weight loss , improving fat loss, and maintaining lean body mass. Derived from Theobroma cacao to obtain the benefits of a first component selected from the group consisting of extracts, fractions, phytochemical components and mixtures thereof; Derived from Citrus aurantifolia a second component selected from the group consisting of extracted extracts, fractions, phytochemical components and mixtures thereof; and a pharmaceutically acceptable excipient, diluent, and carrier, or a combination thereof. The present invention relates to a synergistic herbal composition optionally containing at least one ingredient selected from the group consisting of: The present invention also provides extracts, fractions, phytochemical components and the like derived from Theobroma cacao. A first component selected from the mixture of these compounds and a compound derived from Citrus aurantifolia. a second component selected from the group consisting of extracted extracts, fractions, phytochemical components and mixtures thereof; and a pharmaceutically acceptable excipient, diluent, and carrier, or a combination thereof. A synergistic herbal composition optionally containing at least one ingredient selected from the group consisting of: The use of the product to prevent, control, or reduce obesity and / or overweight in humans Treats and improves lean body mass and enhances browning of white adipose tissue (WAT) / brown adipose tissue Improves the formation of adipose tissue (BAT) and increases basal metabolic rate (BMR) / energy expenditure Stabilizes mass, improves thermogenesis, improves thyroid function, maintains healthy weight, and promotes satiety Choose from increasing blood sugar, aiding in weight loss, enhancing fat loss and maintaining lean body mass. The present invention relates to a method for obtaining at least one health benefit from [Background technology]

[0002] Obesity and overweight are rapidly growing conditions in an increasing number of countries around the world. A recent WHO report stated that global obesity rates have tripled since 1975.30 kg / m 2 A person with a body mass index (BMI) above 100 is generally considered to be overweight. and overweight is 25 kg / m 2 A confirmed BMI of 100 or above Both conditions are caused by an imbalance between energy intake and expenditure. Obesity is associated with substantial increases in morbidity, premature mortality, reduced quality of life, and Excess weight is associated with increased risk of diabetes, metabolic syndrome, Hypertension, dyslipidemia, myocardial infarction, stroke, certain cancers, sleep apnea and osteoarthritis This is a key area for developing therapeutic drugs that can reduce body weight. Key strategies include reducing food consumption or absorption and / or increasing energy expenditure. Many drugs have been commercialized as an answer to the obesity problem, but Most have been withdrawn from the market due to unacceptable side effects. Researchers in the field of metabolic disorders are seeking treatments for obesity / overweight and related diseases, and are looking to address the food-related challenges. These compounds are mainly derived from the plant and marine kingdoms. Some herbal extracts and dietary ingredients may have effects on appetite regulation, fat oxidation, energy intake or thermogenesis. Herbal ingredients have been shown to have few harmful side effects. These are known to have benefits, making them an interesting complementary approach to the management of obesity. may present with roaches.

[0003] Patent Document 1 discloses Alchemilla vulgaris, Olea europaea, Cuminum cyminum num cyminum) and Mentha longifolia (Mentha longif lora) mixture (weight ratio 12:10:5:4) and Theobroma cacao, caffeine Anhydrous Coffea Arabica, Coffea Canef Coffea canephora, Camellia sinensis sinensis), Ilex paragua riensis), guarana, and kona is doing.

[0004] Patent document 2 discloses the use of theobroma for treating receptor tyrosine kinases associated with disorders. -Discloses cocoa extract.

[0005] Patent document 3 targets psoriasis and other skin diseases such as acne and fungal diseases. Cucumis sativus, Citrus aurante Pimenta racemosa and Clapton Aqueous liquid extraction of Graptophyllum pictum leaves The present invention discloses a composition containing the substance.

[0006] Patent Document 4 discloses at least one fermented food product containing catechin and theobroma cacao extract as a source. It discloses a dietary supplement containing 3% corosolic acid to promote weight loss.

[0007] Patent Document 5 discloses Zingiber officinale ), Cotinus coggygria, Citrus o Citrus aurantium, Lupron, Whey Protein Monohydrate Substances, chromium polynicotinate, hexahydroisoalpha-acid, xanthohumol, rohi Soybean alpha acids, elderberry, Gymnema sylvestre ), Camellia sinensis, Acacia nilotica , Malus pumila, Ribes nigrum L igrum L), Hypericum perforatum (Hypericum perfo ratum), Theobroma cacao, Vaccinium myrtillus, Camellia sinensis, Rosa canina ( Rosa canina), isoalpha acids, vaccinium erythrocarpam (Vac cinium erythroCarpum), leucine, Goldenseal canadensis (Hydrastis Canadensis), Vitis vinifera (Viti s vinifera), Rhamnus purshiana ), Epimedium (Horney Gorgeweed), Curcuma longa (Curcuma longa), Opuntia ficus indica (Opuntia ficus indica ndica), Syzygium cumini and Tetrahymena Phytochemical constituents or extracts isolated from dholoisoalpha-acids are included in the group of pharmaceutically active compounds. By using a composition containing one or more components in an amount, AMP, which is an elastic cell in an animal, can be A method for activating K is disclosed.

[0008] Another patent document 6 discloses a compound containing effective amounts of green tea, ginger, caffeine, cocoa, calcium, and yam. Ruba Mate, Hawthorn Berry, Parsley Leaf Marshmallow Root, Fennel Seed, Astragalus root, licorice root, soybean, cinnamon, celery seed and alfalfa leaf, The present invention discloses a dosage form of tablet.

[0009] Therefore, alternatives containing highly effective herbal compositions for the management of obesity and overweight are available. There is a continuing need in the art to provide effective treatments. Herbal extracts and compositions that are well tolerated and more effective than ever before are useful in treating obesity and There is an urgent need for a review of overweight. [Prior art documents] [Patent documents]

[0010] [Patent Document 1] U.S. Patent Publication No. 2016 / 045560(A1) [Patent Document 2] International Publication No. WO2016 / 046375(A1) [Patent Document 3] International Publication No. WO2008 / 110853(A1) [Patent Document 4] Australian Patent Publication No. 2006312947(B2) [Patent Document 5] U.S. Patent Publication No. 2010 / 0112099(A1) [Patent Document 6] U.S. Patent No. 7,329,419 (B2) Summary of the Invention [Problem to be solved by the invention]

[0011] Object of the invention The object of the present invention is to prevent, control, or treat obesity and / or overweight and to increase lean body mass. Improves weight and promotes browning of white adipose tissue (WAT) / brown adipose tissue (BAT) Improves metabolism, increases basal metabolic rate (BMR) / stabilizes energy expenditure, and reduces fever Increases blood sugar production, improves thyroid function, maintains a healthy weight, increases satiety, and supports weight loss and at least one health benefit selected from: promoting fat loss and maintaining lean body mass. Extracts, fractions, and phytochemicals derived from Theobroma cacao for health benefits and mixtures thereof, and a first component selected from Citrus aurantifolia or A second component selected from extracts, fractions, phytochemical components and mixtures thereof derived from the plant. The present invention provides a synergistic herbal composition comprising a combination of

[0012] Another object of the present invention is to provide a method for preventing and controlling obesity and / or overweight in humans or animals. Control or treat to improve lean body mass and enhance browning of white adipose tissue (WAT) / Improves brown adipose tissue (BAT) formation and increases basal metabolic rate (BMR) / Energy Stabilizes ghee consumption, improves thermogenesis, improves thyroid function, maintains healthy weight and promotes satiety. Increases abdominal tone, aids weight loss, and provides a way to improve fat loss and maintain a lean body The method comprises the steps of: preparing extracts, fractions, phytochemicals, and the like derived from Theobroma cacao; a first ingredient selected from the group consisting of Citrus aurantifolia, Citrus aurantifolia, and a mixture thereof; a second extract, fraction, phytochemical component, or mixture thereof derived from the plant; The present invention includes supplementing a human with an effective amount of a synergistic herbal composition comprising a combination of ingredients of:

[0013] Yet another object of the present invention is to provide a method for preventing, controlling, or treating obesity and / or overweight, Improves lean body mass and promotes browning of white adipose tissue (WAT) / brown adipose tissue (B Improves the formation of ATP, increases basal metabolic rate (BMR) and stabilizes energy expenditure Increases thermogenesis, improves thyroid function, maintains a healthy weight, increases satiety, At least one of the following is selected to aid in weight loss, enhance fat loss, and maintain lean body mass: Extracts, fractions, and botanicals derived from Theobroma cacao to obtain several health benefits. a first component selected from the group consisting of citrus aurantium nitrite and mixtures thereof; The present invention relates to a method for producing a plant extract, a plant fraction, a plant chemical component, and a mixture thereof, which is selected from the group consisting of extracts, fractions, phytochemical components, and mixtures thereof derived from the plant extract, The present invention provides for the use of a synergistic herbal composition comprising a combination of two ingredients. [Means for solving the problem]

[0014] Summary of the Invention The present invention provides a method for preventing, controlling, or treating obesity and / or overweight and improving lean body mass. Improves the browning of white adipose tissue (WAT) and promotes the formation of brown adipose tissue (BAT) Improves metabolism, increases basal metabolic rate (BMR), stabilizes energy expenditure, and increases thermogenesis Improves thyroid function, maintains a healthy weight, increases satiety, and aids in weight loss At least one health benefit selected from: Extracts, fractions, phytochemicals and other compounds derived from Theobroma cacao to obtain the above benefits and a first component selected from Citrus aurantifolia and mixtures thereof. a second component selected from derived extracts, fractions, phytochemical components and mixtures thereof; The present invention provides a synergistic herbal composition comprising a combination of:

[0015] Another aspect of the present invention is a method for preventing, controlling, or treating obesity and / or overweight in humans. Treat and improve lean body mass and enhance browning of white adipose tissue (WAT) / brown fat Improves the formation of BAT tissue and increases basal metabolic rate (BMR) / energy expenditure stabilizes blood sugar, improves thermogenesis, improves thyroid function, maintains a healthy weight, and promotes satiety Choose from increasing blood cholesterol, supporting weight loss, enhancing fat loss and maintaining lean body mass A method for obtaining health benefits from an extract derived from Theobroma cacao is provided. a first component selected from the group consisting of Citrus aurantifolia, ... A combination of a second component selected from extracts, fractions and mixtures thereof derived from the plant. and (iii) administering to a human an effective amount of a synergistic herbal composition comprising: and optionally at least one component selected from excipients, diluents, and carriers thereof. This includes.

[0016] Another aspect of the present invention is a method for preventing, controlling, or treating obesity and / or overweight in humans. Treat and improve lean body mass and enhance browning of white adipose tissue (WAT) / brown fat Improves the formation of BAT tissue and increases basal metabolic rate (BMR) / energy expenditure stabilizes blood sugar, improves thermogenesis, improves thyroid function, maintains a healthy weight, and promotes satiety Choose from increasing blood cholesterol, supporting weight loss, enhancing fat loss and maintaining lean body mass , an extract derived from Theobroma cacao to obtain at least one health benefit. a first component selected from the group consisting of Citrus aurantifolia, ... A combination of a second component selected from extracts, fractions and mixtures thereof derived from the plant. The present invention provides for the use of an effective amount of a synergistic herbal composition comprising: Optionally, it comprises at least one component selected from diluents, and carriers thereof.

[0017] Another aspect of the present invention is the preparation of extracts, fractions and their derivatives derived from Theobroma cacao. A first component selected from the mixture and an extract derived from Citrus aurantifolia. and a second component selected from the group consisting of a mixture of the extract, a fraction, and a mixture thereof. providing a synergistic herbal composition and improving metabolic processes, e.g., promoting lipolysis; and inhibition of adipogenesis, increase in fibroblast growth factor-21 (FGF-21), uncoupling protein Increased levels of protein (UCP-1) and increased levels of β3-adrenergic receptors (β3-ARs) and a carrier thereof, Optionally, the composition may contain at least one component. [Brief explanation of the drawings]

[0018] [Figure 1]1A is a bar graph showing the body weights of animals in the control and treatment groups on days 1, 8, 15, 22, and 28. Each bar represents the mean body weight ± SM. p<0.05 indicates significance, # indicates G1 vs. G2, and * indicates G2 vs. treatment group. The bar graph shows the percentage change in body weight of the control and treatment groups of animals on day 28 (B). G1 and G2 represent groups of rats supplemented with normal chow and high-fat chow, respectively. G3, G4, and G5 represent rats supplemented with 100 and 300 mg / kg body weight of Composition-67 and high-fat chow supplemented with 10 mg / kg body weight of sibutramine, respectively. [Figure 2] The bar graph shows the reduction in daily dietary calorie intake by diet-induced obese rats supplemented with Composition 67. Each bar represents the mean ± SD. G1 and G2 represent groups of rats supplemented with normal diet and high-fat diet. G3, G4, and G5 represent rats supplemented with 100 and 300 mg / kg body weight of Composition 67 and high-fat diet supplemented with 10 mg / kg body weight of sibutramine, respectively. n=7, p<0.05 significant, # is G1 vs. G2, * is G2 vs. treatment group. [Figure 3] The bar graph shows the reduction of visceral fat mass in diet-induced obese rats supplemented with Composition 67. Each bar represents the mean ± SD. G1 and G2 represent groups of rats supplemented with normal diet and high-fat diet. G3, G4, and G5 represent rats supplemented with 100 and 300 mg / kg body weight of Composition 67 and high-fat diet supplemented with 10 mg / kg body weight of sibutramine, respectively. n=7, p<0.05 is significant, # is G1 vs. G2, * is G2 vs. treatment group. [Figure 4]The bar graph shows the change in epididymal adipocyte size in diet-induced obese rats supplemented with Composition 67. Each bar represents the mean ± SE of adipocyte area in μm². G1 and G2 represent groups of rats supplemented with normal diet and high-fat diet, respectively. G3, G4, and G5 represent groups of rats supplemented with 100 and 300 mg / kg body weight of Composition 67 and high-fat diet supplemented with 10 mg / kg body weight of sibutramine, respectively. n=7, p<0.05 significant, # is G1 vs. G2, * is G2 vs. treatment group. [Figure 5] The bar graph shows the normalization of serum leptin levels in diet-induced obese rats supplemented with Composition 67. G1 and G2 represent groups of rats supplemented with normal diet and high-fat diet, respectively. G3, G4 and G5 represent rats supplemented with 100 and 300 mg / kg body weight of Composition 67 and high-fat diet supplemented with 10 mg / kg body weight of sibutramine, respectively. n=7, p<0.05 is significant, # is G1 vs. G2, * is G2 vs. treatment group. [Figure 6] The bar graph shows the improvement of thyroid hormone balance in diet-induced obese rats supplemented with Composition 67. Each bar represents the mean ± SD of the ratio of triiodothyronine (T3) and thyroxine (T4). G1 and G2 represent groups of rats supplemented with normal diet and high-fat diet, respectively. G3, G4, and G5 represent groups of rats supplemented with 100 and 300 mg / kg body weight of Composition 67 and high-fat diet supplemented with 10 mg / kg body weight of sibutramine, respectively. n=7, and the number on each bar represents the percentage of the relative T3 / T4 ratio, which is considered to be 100% in G1. [Figure 7] The photomicrograph shows immunohistochemical staining of UCP-1 in epididymal adipose tissue. G1 and G2 represent groups of rats supplemented with normal diet and high-fat diet, respectively. G3, G4, and G5 represent groups of rats supplemented with high-fat diet supplemented with 100 and 300 mg / kg body weight of Composition 67 and 10 mg / kg body weight of sibutramine, respectively. Bar: 50 μm. DETAILED DESCRIPTION OF THE INVENTION

[0019] Detailed Description of the Invention The present invention will now be described in detail with reference to certain preferred and optional embodiments. so that their various aspects may be fully understood and appreciated.

[0020] The terms "adipollysis" and "lipolysis" are art-recognized terms and are used interchangeably throughout this specification. Those skilled in the art will recognize similar terms as such. Similarly, understand and appreciate the differences between browning, browning of fat, beige fat and white fat. Browning of adipose tissue (WAT) is also used interchangeably. The terms "herb" and "plant" are also used interchangeably. The terms "item" and "item" are also used interchangeably throughout this specification.

[0021] Adipogenesis is the process of differentiation and proliferation of preadipocytes into mature adipocytes or fat cells. In this process, preadipocytes or preadipocytes are transformed into mature adipose tissue. After differentiation into the adipocyte phenotype, they proliferate. The nuclear receptor PPARγ mediates adipocyte differentiation and adipose tissue proliferation. Adipocytes are known to play an important role in fat deposition. It plays an essential role in the metabolism of large amounts of fatty acids, such as triglycerides in animals. Fat cells exist in a dynamic state and they are responsible for maintaining energy reserves. When consumption exceeds intake, hypertrophy begins. This process is It is highly regulated by antagonistic hormones that are always highly sensitive. Harm is one of the main goals of developing treatments for obesity / overweight and promoting weight loss. be.

[0022] Adipolysis (lipolysis) is the breakdown of triglycerides stored in fat cells. This is a metabolic process that promotes the release of fatty acids and glycerol into the bloodstream. is a well-regulated process that allows for the proper delivery of free fatty acids and promotes energy production. Therefore, increasing adipolysis can treat and reduce obesity / overweight. One of the main goals of β3-adrenergic receptor agonists is to promote the It can stimulate lipolysis in colored adipose tissue and thermogenesis in brown adipose tissue. Plant extracts, fractions and phytochemicals with lipolytic activity are effective in treating obesity, overweight and other conditions. and other metabolic disorders.

[0023] Therefore, the inventors of the present application have identified a novel compound with anti-adipogenic and pro-adipogenic activity. We randomly screened many plant extracts and fractions to find the best extracts from Theobroma cacao and The extracts and fractions derived from Citrus aurantifolia are summarized in Tables 3 to 7. As shown in Fig. 1, the effective dose was dependent on anti-adipogenic and preadipocyte activity. I put it out.

[0024] A brief summary of each of the plant materials is provided herein below.

[0025] Theobroma cacao: Theobroma cacao L. is a small but economically important tree. It is an evergreen tree of the Sterculiaceae family, 4-8m tall, native to tropical America. Cacao seeds are an important source of polyphenols and theobromine. Used to make cocoa mass, cocoa powder, confectionery, gouache and chocolate Cocoa extract or its phytochemical components have been shown to reduce platelet aggregation, hypertension, and atherosclerosis. atherosclerosis, hyperglycemia and hypercholesterolemia, inflammation, liver cancer formation, DNA damage Theobromine (I) has shown some beneficial effects on the mitochondrial and clastogenic effects of cerebrospinal fluids. is the principle alkaloid in Theobroma cacao (Donald L. Pavi a,Journal of Chemical Education,1973,50, 791-792) and the extract used in the present invention was found to be theobroma by HPLC analysis. It is standardized in

[0026] [ka]

[0027] Citrus aurantifolia: Citrus aurantifolia grows to a height of 3-5m. It is a perennial evergreen tree that can grow to a height of 100 meters. It has irregularly thin branches and short, It has sharp, stiff thorns or needles. The flowers are short, partly white, and fragrant. The fruit is round, 3-5 cm in diameter, and turns yellow when ripe. Classification of S. aurantifolia: Kingdom (Plantaceae), Inferior Kingdom (Vascular Plants), Superphylum (Seed Plants) ), Phylum (Angiosperms), Class (Magnoliaceae), Subclass (Rosaceae), Order (Sapindales), Family (Milk) Citrus aurantifolia (family Citrus), genus (Citrus), species (Citrus aurantifolia). It is found in India, China It is native to tropical and subtropical regions of Asia and Southeast Asia, including North Africa, The common name for Citrus aurantifolia is lime. (English). It is used not only as a flavoring agent in beverages, processed foods, and dosage forms, but also as a It is also used as an ingredient in perfumes. The peel of Citrus aurantifolia has antioxidant properties. Antidiabetic activity, antilipidemia activity, anti-insecticide activity, anti- It has traditionally been used as a cancer inhibitor, which in vitro inhibits xanthine oxidase activity. It has shown inhibitory, antioxidant, cytotoxic and antiviral effects. Limonin (II) is found in Citrus aurantium bergamia and other cosmetics. The extract of the present invention has been confirmed as a principal metabolite of riboflavin by HPLC analysis. Standardized to monin(II).

[0028] [ka]

[0029] The sources of herbs used in this invention are as follows:

[0030] 1. Theobroma cacao seeds are from Aswaraopeta Village, Aswaraopet a Panchayat, Aswaraopeta District, Bhadradri Kothagud It was collected from cultivated sources in em district, Telangana state.

[0031] 2. Citrus aurantifolia is grown in Rayapudi Village, Rayapudi Pan Chayat, Thullur Upazila, Guntur District, Andhra Pradesh Collected from cultivated sources in the

[0032] Theobroma cacao seeds were crushed and the powder was dissolved in water, aqueous ethanol, ethanol, aqueous methicone, and ethanol. Extraction was performed using various solvents, such as ethanol and n-butanol, and the aqueous extracts were (TC-1), aqueous ethanol extract (TC-2), ethanol extract (TC-3 ), aqueous methanol extract (TC-4) and n-butanol extract (TC-5) Theobroma cacao seed extract was analyzed by HPLC and found to contain theobromine. The results are summarized in Table 1. Similarly, Citrus aurantifolia The fruit peel was crushed and the powder was dissolved in aqueous ethanol, ethanol, water, aqueous methanol and n- Extraction was performed using various solvents such as butanol and aqueous ethanol extracts (CA). -1), ethanol extract (CA-2), water extract (CA-3), aqueous methanol extract The Citrus extract (CA-4), and n-butanol extract (CA-5) were obtained. The extract of L. lanthifolia was standardized for limonin (II) by HPLC analysis. The results are summarized in Table 2.

[0033] Theobroma cacao seed extract and Citrus aurantifolia fruit extract Skin extract was used in an in vitro cell model in mouse 3T3-L1 preadipocyte cells. , were evaluated for their anti-adipogenic & pro-adipogenic activity. He pointed out that it is highly effective in inhibiting lipogenesis and increasing adipolysis. For example, the aqueous extract of Roma cacao seeds (TC-1) was found to be 5 μg / mL and 10 μg / mL. At the treatment concentration of 1 mL, the adipogenesis inhibition was 38.36% and 41.10%, respectively. The aqueous ethanol extract of Citrus aurantifolia fruit peel (CA-1) At treatment concentrations of 5 μg / mL and 10 μg / mL, 25.81% and 35.74% Theobroma cacao seeds and Citrus au Lait showed inhibition of adipogenesis, respectively. Other solvent extracts of the peel of the Tifolia fruit have also been found to be effective.

[0034] These individual extracts or fractions thereof are then combined to determine the synergistic effectiveness of these components. Theobroma cacao extracts or fractions were evaluated to investigate their potential. Extracts or fractions derived from Citrus aurantifolia and Citrus aurantifolia were prepared in different ratios. The compositions (Compositions 1 to 65) were obtained by mixing the ingredients in the above ratio. 65) was compared with the corresponding individual components to test their inhibitory effects on adipogenesis. Data from an in vitro adipogenesis inhibition assay for compounds derived from Theobroma cacao The derived extract(s), or fraction(s), or phytochemical constituents, or mixtures thereof The compound may be an extract(s) or fraction(s) derived from Citrus aurantifolia. or phytochemical constituents or mixtures thereof that exhibit synergistic properties when combined with The results suggest that the inhibitors of adipogenesis may be more pronounced when compared to their corresponding individual components. showed unexpectedly good efficacy in terms of harm.

[0035] For example, the water extract of Theobroma cacao seeds (TC- 1) and 50% of Citrus aurantifolia fruit peel at a concentration of 3.33 μg / mL % aqueous ethanol extract (CA-1) increased adipogenesis by 15.23% and 19.47% Theobroma cacao seeds at 5 μg / mL and a 1:2 ratio showed inhibition of (TC-1) and 50% aqueous extract of Citrus aurantifolia fruit peel Composition-2 containing the ethanol extract (CA-1) inhibited adipogenesis by 44.78%. This was 34.7% calculated from the inhibition exhibited by the corresponding individual components. This is significantly better than the additive effect of these two extracts (15.23% + 19.47%). The products (TC-1 and CA-1) were mixed at 1:3, 1:1, 2:1 and 3:1 ratios, respectively. Composition-1 (C-1) and Compositions-3 to 5 (C-1 and C-3) containing the following ratios: ~C-5) also vary with the corresponding individual component concentrations, as summarized in Table 3. When compared with the inhibition shown, synergy was demonstrated. of Theobroma cacao seeds in combination with other solvent extracts (CA-2 to CA-5) Other compositions (C6-C19) containing the water extract (TC-1) also exhibit synergistic lipid The results showed inhibition of lipogenesis (Table 3).

[0036] An additional example is the 50% aqueous ethanol solution of Theobroma cacao seeds at a concentration of 0.83 μg / L. Nol extract (TC-2) and Citrus aurantifolia at a concentration of 1.67 μg / L The 50% aqueous ethanol extract of the peel of Olia (CA-1) showed 2.07% and 26.4% At 2.5 μg / mL, Theobroma cacao inhibited lipogenesis by 6%. 50% aqueous ethanol extract of seeds (TC-2) and Citrus aurantifolia A 50% aqueous ethanol extract of barley peel (CA-1) was added in a 1:2 ratio. Ingredient-21 showed 40.19% inhibition of adipogenesis, which was comparable to the corresponding individual ingredients. The additive effect of 28.53% (2.07% + 26.46%) was calculated from the inhibition shown by These two extracts (TC-2 and CA-1) were significantly better than other Composition-20 and Composition-22 to Composition-24 (C-20, C-2) containing the following component ratios: 2-C-24) and Citrus aurantifolia fruit peel and other solvent extracts. Contains a 50% aqueous ethanol extract of combined Theobroma cacao seeds (TC-2) The compositions also show the corresponding individual component concentrations as summarized in Table 4. This demonstrated synergy when compared to the inhibition shown by

[0037] Similarly, other solvent extracts of Theobroma cacao and Citrus aurantifolia Compositions (C-39 to C-65) containing the above-mentioned compounds have the corresponding individual component concentrations as summarized in Table 5. The synergy was demonstrated when compared to the inhibition exhibited by each of the two.

[0038] The compositions (Compositions 1-65) showed a significantly improved response to 3T3-L1 IgG in mice compared to the corresponding individual components. Efficacy of increasing adipolysis using an in vitro cell model in preadipocyte cells Further testing was performed on these compositions. Theobroma cacao extract(s) or fraction(s) or The phytochemical constituents or mixtures thereof are derived from Citrus aurantifolia Extract(s), or fraction(s), or phytochemical constituents or mixtures thereof Their corresponding individual components have been shown to have a tendency to exhibit synergy when mixed. showed unexpectedly good efficacy in terms of increasing adipolysis when compared with 1 min.

[0039] For example, the water extract of Theobroma cacao seeds (TC- 1) and 5) of Citrus aurantifolia fruit peel at a concentration of 13.33 μg / mL. The 0% aqueous ethanol extract (CA-1) increased the adipose tissue activity by 29.52% and 7.12%. At 20 μg / mL, the lysis of Theobroma caca at a ratio of 1:2 was increased. Water extract of Cao seeds (TC-1) and Citrus aurantifolia peel 50 Composition-2 containing 47.88% aqueous ethanol extract (CA-1) showed an increase in polysulfones, calculated from the increases shown by the corresponding individual components. This is significantly better than the additive effect of 36.64% (29.52% + 7.12%). The composition contains two extracts (TC-1 and CA-1) in different ratios. Compositions 1 and 3 to 5 also show the corresponding individual component concentrations as summarized in Table 6. Theobroma cacao water extract showed synergy when compared to the increases shown by the other extracts. (TC-1) and Citrus aurantifolia (CA-2 to CA-5) Other compositions (C6-C19) containing other solvent extracts also exhibited synergistic pro- active effects. Theobroma cacao and Citrus au Lait showed polycystic activity (Table 6). Compositions containing other solvent extracts of T. tifolia (C-20 to C-65) are summarized in Table 7. Thus, synergy is observed when compared to the increases exhibited by each of the corresponding individual component concentrations. was presented.

[0040] Interestingly, Theobroma cacao seeds and Citrus aurantifolia fruits Extracts and fractions derived from fruit skins and compositions thereof have been shown to be effective in, for example, browning fat, Thermogenesis, stabilization of energy expenditure, improved thyroid function, increased satiety, discussed below These factors, such as FGF21, UCP-1, and β3-AR, contribute to the browning of white adipocytes. Other mechanisms for investigating obesity and overweight, such as regulating contributing molecular factors, are also being explored. The study further demonstrated unexpected efficacy in

[0041] Browning of brown adipose tissue (BAT), white adipose tissue (WAT) and energy expenditure Stabilization: Adipose tissue (body fat) is a loosely bound connective tissue composed of fat cells. In humans, adipose tissue is mainly located under the skin (subcutaneous adipose tissue). White adipose tissue (WAT) is primarily located in the lining of the body and around the internal organs (visceral fat). ), also known as white fat, is one of two types of adipose tissue found in mammals. The other type of adipose tissue is brown adipose tissue (BAT). They store energy in the form of fat, which causes pathological hypertrophy during obesity. Pigmented adipose tissue (BAT) is a unique form of adipose tissue in humans and mammals. T is able to dissipate large amounts of chemical energy as heat through a process called thermogenesis. BAT is not only an important biological defense against hypothermia, but also a key regulator of metabolism through eating and drinking. Brown fat cells also play a role in the production of heat induced by thermogenesis. It contains a unique protein known as uncoupling protein 1 (UCP-1). UCP-1 is a short-term circulator of the electron transport chain, but is otherwise a It is normally used to drive cellular ATP synthesis, which allows the mitochondrial membrane The potential makes BAT capable of transferring heat (thermogenesis), which allows it to Compositions that can alter energy expenditure and fuel metabolism without increasing It becomes a weave.

[0042] Recently, the topic of brown adipose tissue has been expanding, as is known from the browning of WAT, and the browning of white adipose tissue ( The study is being rekindled by a number of new studies on the conversion of WAT (whole body fat) to beige fat. WAT, derived from several depots, responds to appropriate stimuli and This can trigger a process known as "browning" for the production of erythrocytes. The cells contain multivesicular lipid droplets and multiple mitochondria, and specific proteins such as UCP-1 are expressed in the cells. Acquire BAT properties by increasing protein expression. "Browning" stimulus Beige adipocytes induced by α-glucan have significant amounts of mitochondria and UCP-1. These cells are phenotypically similar to the classical brown adipocytes in BAT. It has been suggested that adipocytes may have the same thermogenic capacity as BAT. FGF21 plays a physiological role in the thermogenic mobilization of WAT. It is also an important regulator of browning. PGC1α is a key transcriptional central regulator of WAT browning The browning of white adipose tissue is also mediated by the action of norepinephrine and β3 -Adrenergic receptors (β3AR) are involved, and sympathetic stimulation β3AR activation drives the expression of UCP-1 and other thermogenic proteins in white adipocytes. This sequence of events initiates a signaling cascade that leads to the overexpression of proteins. is a brown adipocyte of white fat cells that has undergone intermediate stages such as beige or brite cells. In addition, thyroid hormones, especially T3, positively regulate UCP-1 synthesis. T3 stimulates UCP-1 synthesis individually or in a synergistic manner, acting in concert with sympathetic It can function in conjunction with stimulation. The activity of T3 is inhibited by T4. Therefore, a healthy balance of T3 and T4 is important for the browning of white adipocytes.

[0043] FGF-21: Fibroblast growth factor-21 (FGF-21) is a new member of the FGF family. FGF-21 is a key member of the FGF family and is predominantly expressed in the liver. Regulates stasis, insulin sensitivity, ketogenesis, and promotes adipose tissue "browning" Over the past decade, FGF-21 has been shown to be a key regulator of obesity and diabetes. Adipose tissue has been shown to be a potential therapeutic target for the treatment of adipose tissue diseases. One of the primary targets of action and one of the key metabolic benefits of FGF-21 is "browning." New evidence supports the role of UCP-1 and other thermoregulatory factors in the autocrine-paracrine system. FGF-21 promotes thermogenesis and white adipose tissue metabolism by upregulating the expression of metabolic genes. These results suggest that FGF21 plays a physiological role in the thermogenic mobilization of fat. Increases PGC1α levels in adipose tissue.

[0044] Some compositions selected from Composition-1 to Composition-65 are used to treat 3T3- Fibroblast growth factor-21 (FGF-21) in a cell assay in L1 preadipocyte cells These compositions were tested for their ability to increase the Data from an in vitro FGF-21 assay derived from Theobroma cacao The extracted extract(s), or fraction(s), or phytochemical constituents or mixtures thereof , extract(s), or fraction(s) derived from Citrus aurantifolia tendency to exhibit synergistic effects when mixed with other plant chemicals or their mixtures. The increase in FGF-21 and its related effects on the immune system when compared to their respective individual components suggests that The results showed unexpectedly good efficacy in this regard.

[0045] For example, the water extract of Theobroma cacao seeds (TC- 1) and 50% of Citrus aurantifolia fruit peel at a concentration of 3.33 μg / mL % aqueous ethanol extract (CA-1) contained 10.08% and 18.60% FGF- At 5 μg / mL, theobroma cacao at a 1:2 ratio showed an increase of 21%. Water extract (TC-1) and 50% aqueous ethanol of Citrus aurantifolia Composition-2 containing the extract (CA-1) showed an increase of 44.21% in FGF-21. This was calculated from the increase exhibited by the corresponding individual components by 28.68% (1 This is significantly better than the additive effect of 0.08% + 18.60%). Composition-1 and Composition-2 containing two extracts (TC-1 and CA-1) -3 to 5 are also indicated by the corresponding individual component concentrations, as summarized in Table 8. The inhibition of the TC-10001 was synergistic when compared with that of the TC-100011. 1) and other solvent extracts of Citrus aurantifolia (CA-2~CA-5 ) and other compositions containing C-10 and C-15 fatty acids, as well as Theobroma cacao. Compositions containing other solvent extracts of the seeds and peel of Citrus aurantifolia -20 to 23 and compositions -36 to 38 (C-20 to C-23 & C36 to 38) also The increase exhibited by each of the corresponding individual component concentrations is summarized in Table 8. showed a synergistic increase in FGF21 activity when compared with the addition of

[0046] UCP-1: The global epidemic of obesity and other metabolic disorders has recently prompted scientists to focus on white fat The focus has shifted to adipose tissue (WAT) and its biology. WAT is responsible for certain physiological functions. and under pathophysiological conditions, can convert to brown adipose-like tissue. Tissue depots significantly increased gene expression for uncoupling protein 1 (UCP-1), which As a result, white fat "browning" occurs, which is generally thought to have thermogenic and fat-burning properties. This phenomenon has attracted considerable attention in changing its function from energy storage to energy dissipation. UCP-1 is an integral membrane protein found in the inner mitochondrial membrane of brown adipose tissue. It is a toxic protein that aids in the process of non-shivering heat production in mammals. Therefore, the researchers' primary focus is on eliciting the "browning" response and increasing the U of adipose tissue. The goal is to identify agents that increase CP-1 expression and activity, which ultimately contributes to obesity and It combats the development of obesity and overweight as well as other metabolic disorders and promotes energy expenditure.

[0047] Therefore, some of the compositions selected from Compositions 1 to 65 are useful for the treatment of mouse 3T3 -L1 preadipocyte cells were tested for uncoupling protein-1 (UCP-1) expression activity. Data from in vitro UCP-1 assays for these compositions were unexpected. showed a significant increase in UCP-1 expression over the control. Theobroma cacao seed water extract (TC- 1) and a 50% aqueous ethanol extract of the peel of Citrus aurantifolia fruit (C Compositions 1 to 5 containing .A-1) exceeded the control by 38.48% and 34.23%. , with significant increases in UCP-1 expression of 33.81%, 30.71%, and 28.69%. Similarly, theobroma cacao seeds at 1:1 and 2:1 ratios showed high efficacy. 50% methanol extract (TC-4) and Citrus aurantifolia fruit peel Compositions 58 and 59 containing a 50% aqueous ethanol extract (CA-1) of Significant increases in UCP-1 expression were observed over the control group, with increases in UCP-1 expression of 24.63% and 11.96%. Therefore, the data summarized in Table 9 show that Theobroma cacao Extracts or fractions derived from Citrus aurantifolia and extracts or fractions derived from Citrus aurantifolia A composition containing the extract or fraction may have the potential to upregulate UCP-1 expression, and Increased adipose browning, thermogenesis, and stabilization of energy expenditure were correlated with increased adipose browning in the presence of steroids. This suggests that:

[0048] β3AR: β3-AR is a β-adrenergic receptor located primarily in adipose tissue. Its main functions include regulation of lipolysis and thermogenesis. Classical BAT stores are , which are highly innervated and respond to, for example, cold exposure and certain chemicals in the diet. It is activated by centers in the brain in response to specific stimuli, such as exposure. It promotes the release of norepinephrine (NE) from nerves. NE is released by the BAT β3-adrenergic receptor. When it binds to the β3-AR, it increases the level of intracellular cyclic AMP (cAMP). The addition of niacin promotes lipolysis, and the breakdown of triglycerides results in the production of uncoupling proteins. Activation of UCCP-1 upregulates and activates UC protein-1, resulting in the release of free fatty acids. P-1 uncouples mitochondrial respiration, which generates thermogenesis, thereby increasing BAT lipid In adipocytes, β3-AR signaling is conspicuously enriched in mitochondria, and it is essential for the respiration In addition to classical brown adipocytes (BAT), beige or or bright adipocytes may also have a similar effect. These are induced by various stimuli, most notably exposure to cold or β-AR signaling. by activators of these proteins and by peroxisome proliferator-activated proteins such as rosiglitazone. Can be "browned" by receptor (PPARγ) agonists. Bright / beige adipocytes Activation of mitochondrial ATP also results in an increase in uncoupled mitochondria, similar to the occurrence in BAT. Therefore, activation of β3-adrenergic receptors (ARs) induces browning, thermogenesis, and This may result in a stabilization of energy expenditure (REE) and an increase.

[0049] Therefore, some representative compositions selected from Compositions 1 to 65 were administered to adipocytes. We examined whether β3-adrenergic receptor (β3AR) expression was increased in the rats. In the assay, 3T3-L1 cells treated in vitro with the compositions showed the following results: As summarized in Table 1, unexpectedly, there was a significant increase in β3AR expression in cells treated with β3AR over control-treated cells. For example, theobroma cacao water extracts at ratios of 1:3, 1:2, and 1:1 showed significant increases. Extract (TC-1) and 50% aqueous ethanol extract of Citrus aurantifolia peel Compositions 1, 2 and 3 (C-1, C-2 and C- 3) showed 36.17%, 37.47% and 37.47% increases in β3AR expression over controls, respectively. Similarly, theobroma cacao at 1:2 and 1:1 ratios showed an increase of 51.02%. 50% ethanol extract (TC-2) and Citrus aurantifolia 50% water Compositions-21 & 22 (C-21 & C-) containing ethanol extract (CA-1) 22) showed a 1.5% increase in β3AR expression over controls, as summarized in Table 10. These results were consistent with those of Citrus Aurante. Extracts derived from Theobroma cacao combined with extracts derived from Cacao spp. The composition containing the extract can increase the expression of β3AR, as well as promote adipose browning, This suggests that it may have the ability to increase thermogenesis and lipolysis.

[0050] Dosage Forms: The present invention also provides extracts, fractions and phytochemicals derived from Theobroma cacao. and at least one ingredient selected from the group consisting of Citrus Au Lait, Citrus Oran ... Extracts, fractions and phytochemical components or mixtures thereof derived from Prunus tifolia and optionally a selected second ingredient, pharmaceutical or nutraceutical or nutritional. It contains at least one component selected from acceptable excipients, carriers, and diluents. The present invention provides a synergistic herbal composition.

[0051] The composition prevents, controls or treats obesity, overweight, improves lean body mass, Extracts derived from Theobroma cacao for weight maintenance and lean body mass and at least one component selected from the group consisting of a plant fraction, a plant chemical component, and a combination thereof. Extracts, fractions and phytochemical constituents derived from Citrus aurantifolia and a second ingredient selected from the mixture thereof, and At least one component selected from a nutritionally acceptable excipient, a carrier, and a diluent is optionally added. Optionally, a pharmaceutically, nutraceutical, or nutritionally acceptable excipient or carrier and diluents are monosaccharides such as glucose, dextrose, fructose, and galactose. Sucrose, maltose, lactose, lactulose, trehalose, cellobiose disaccharides such as, but not limited to, chitobiose; starch and starch glycolates; Modified starches such as sodium phosphate, pregelatinized starch, and soluble starch; and Other polycarbohydrates such as modified starches; yellow dextrin, white dextrin, maltodextrin Dextrins and other dextrins produced by hydrolysis of starch or glycogen sorbitol, mannitol, inositol, xylitol, isomalt, etc. polyhydric or sugar alcohols, including but not limited to; microcrystalline cellulose, hydrolyzed Examples include, but are not limited to, hydroxypropyl methylcellulose, and hydroxyethyl cellulose. Not suitable for use in cosmetics, cellulose-based derivatives; Neusilin, Veegum, Talc, Colloidal Dioxide Silicates such as, but not limited to, silicon; calcium stearate, stearin metal stearates, such as, but not limited to, magnesium stearate, zinc stearate Salts; organic acids such as citric acid, tartaric acid, malic acid, succinic acid, lactic acid, and L-ascorbic acid; Polysorbates, fatty acid esters and esters, acacia, carrageenan, guar Natural gums, such as, but not limited to, gum, xanthan gum; B vitamins, nicotine amide, calcium pantothenate, amino acids, casein, gelatin, pectin, agar Proteins such as, but not limited to; sodium chloride, calcium chloride, phosphate Organic metal salts such as, but not limited to, dicalcium, zinc sulfate, zinc chloride; natural face Aqueous solutions of ingredients, fragrances, Class I & II preservatives and the above listed ingredients, alone or in combination; The solution is selected from an alcohol solution, a hydroalcohol solution, and an organic solution.

[0052] For example, Composition-66 contains 60 g of a water extract of Theobroma cacao seeds (TC-1 ), 30g of 50% aqueous ethanol extract of Citrus aurantifolia fruit peel ( CA-1), prepared by mixing 9g of maltodextrin and 1g of syloid. Similarly, Composition-67 was prepared by dissolving 53.33 g of Theobroma cacao in the presence of ethanol / water. Cacao seed water extract (TC-1), 26.67g of Citrus aurantifolia 50% aqueous ethanol extract of the peel of the fruit (CA-1), 18g of maltodextrin and 2 g of syloid were mixed and then dried. was obtained by subjecting 53.2 g of Theobroma cacao seed water extract (T C-1), 26.6g of Citrus aurantifolia fruit peel in 50% aqueous ethanol extract (CA-1), 18.2g of Glusidex-12D and 2g of Syloid The above ingredients were mixed and then dried to obtain a composition.

[0053] In vivo evaluation of Composition-67 in high-fat diet (HFD)-induced obese animals: In vitro In the model, the composition includes Theobroma cacao and Citrus aurantifolia The highly potent anti-obesity and synergistic effects demonstrated by the compound were demonstrated in an in vivo model of obesity. Obesity was further evaluated by feeding male Sprague-Dawley rats a high-fat diet for 4 weeks. The control group (G1) was fed normal chow. After a 4-week induction period, the obese rats were divided into groups of 7 animals each, and then treated with various G2-G3 diets. The animals in the treatment groups were randomly assigned to the G4 group for an additional 4 weeks. Each was administered at 100 mg / kg CMC (G3) or 300 mg CMC (G4) in 10 mL of 0.5% CMC in water. g / kg (G4) body weight composition - 67 or 10 mg sibutramine (G5) orally daily The control group of animals (G2) was supplemented with vehicle (10 mL of 0.5% CMC in water). The weights of individual animals were recorded weekly, and the mean body weight of animals in each group was calculated. The weight gain / change was compared with the initial weight of each animal 1 week after the start of treatment. The mean age was 18 years and the mean age was 18 years. ... In comparison, Composition-67 significantly and dose-dependently reduced the risk of obesity in high-fat diet-induced obese rats. At the end of the treatment period, 100 mg / kg and 300 mg / kg 99.5% and 109% of the body weight in the treatment groups supplemented with Composition-67, respectively. showed a 0.1% decrease in body weight gain, except in the 300 mg / kg dose group. The effect began to show significance after 2 weeks of treatment. The results of body weight and final proportional body weight changes are summarized in Figure 1A and Figure 1B. These results clearly suggest that Composition-67 has potent anti-obesity activity.

[0054] In addition, for rats in groups G3 and G4 supplemented with Composition 67, The average daily food intake was significantly reduced. Food consumption data are shown in Figure 2. These observations are presented as the amount of chlorine in the blood (KJ / day). Oral feeding reduced weight gain and helped maintain a healthy weight in HFD rats, and It has been shown to limit calorie consumption from dietary intake.

[0055] At the end of the study, animals were subsequently euthanized and their viscera (retroperitoneal, epididymal, perirenal and mesenteric) were removed. ) Adipose tissue was collected from rats and weighed. The composition was significantly different when compared to HFD-fed rats. The total fat mass of the group receiving Composition-67 was significantly reduced, and the 300 mg of Composition- The 67(G4) supplemented group showed statistical significance (Fig. 3). In addition, HF D Formalin-fixed paraffin-embedded adipose tissue compared with control group (G2) rats Microscopic examination of the group G4, which was supplemented with 300 mg of Composition-67 for 28 days, showed that It was revealed that the serotonin-dependent ATPase inhibitor effectively reduced fat cell size (Figure 4). The findings showed that supplementation with Composition-67 significantly reduced body fat in obese or overweight rats. This shows that.

[0056] After 28 days of addition, serum samples were analyzed for circulating leptin levels. As summarized in Figure 5, the results of the treatments supplemented with Composition-67 were significantly higher than those of the HFD rats. Interestingly, leptin levels in the treatment groups were significantly reduced. The data show that Composition-67 was effective in reducing satiety and reducing the saturation level to that of chow-fed control animals. showed promising roles in regulating food consumption by regulating the This observation suggests that rats supplemented with Composition-67 in the study consumed significantly less food. The rationale for the reduction can be explained.

[0057] In addition, one interesting observation was that the Composition-67 supplemented rats showed significantly higher GI than the HFD rats. The rats that received this supplement showed improved T3 / T4 balance (Figure 6). It is the active thyroid hormone formed by metabolic conversion from its precursor, T4. , the primary regulator of energy homeostasis in the body, which is responsible for the body's basal metabolic rate Furthermore, T3 also positively influences the stabilization of white fat mass and energy expenditure. It is also an important factor in the conversion process of fat cells into brown fat cells. In overweight animals, the T3 / T4 ratio is lower than in non-overweight individuals. In a study, rats supplemented with Composition-67 had a higher T3 / T4 than HFD rats. This observation suggests that the herbal composition may improve thyroid function in overweight rats. In summary, this observation indicates that supplementation with Composition-67 improved It is strongly suggested that the above-mentioned thyroid function-related brown fat production was induced. These events increased the stabilization of energy expenditure in overweight rats. This provided evidence that body fat was reduced by doing so.

[0058] Immunohistochemistry in epididymal adipose tissue showed that in rats supplemented with Composition-67 The presence of UCP-1 stained brown adipocytes (Fig. 7). The rats fed a high-fat diet (G1) or HFD rats (G2) contained brown fat in their epididymal adipose tissue. This observation therefore suggests that Composition-67 may be involved in the development of brown fat cells. This clearly indicates that the production was potentially induced.

[0059] The foregoing (forgoing) content is an extract, fraction derived from Theobroma cacao a first component selected from phytochemical components and mixtures thereof; and a Citrus Au Lang. Extracts, fractions, phytochemical constituents and mixtures thereof derived from Prunus tifolia The synergistic herbal composition, including a combination of a second ingredient, inhibits adipogenesis and promotes lipolysis. It accelerates the process, increases the production of FGF21, and increases the expression of UCP-1 and β3-AR. , has been shown to increase fat browning and improve T3 / T4 balance. The composition prevents, controls, or treats obesity and / or overweight and promotes lean body mass. Improves the browning of white adipose tissue (WAT) and the formation of brown adipose tissue (BAT) Improves growth, increases basal metabolic rate (BMR) / stabilizes energy expenditure, and promotes heat production Improves vitality, improves thyroid function, maintains a healthy weight, increases satiety, and promotes weight loss At least one of the following will be chosen to help improve fat loss and maintain lean body mass: It can be beneficial for obtaining health benefits.

[0060] Thus, an important embodiment of the present invention is a method for preventing and controlling obesity and / or overweight. Control or treat to improve lean body mass and enhance browning of white adipose tissue (WAT) / Improves brown adipose tissue (BAT) formation and increases basal metabolic rate (BMR) / Energy It stabilizes ghee consumption, improves thermogenesis, improves thyroid function, and helps maintain a healthy weight. Increases satiety, aids weight loss, improves fat loss and helps maintain lean body mass Selected products derived from Theobroma cacao to provide at least one health benefit a first component selected from the group consisting of extracts, fractions, phytochemical components and mixtures thereof; Extracts, fractions, phytochemical constituents derived from Trus aurantifolia and their and a second ingredient combination selected from the mixture of:

[0061] In another embodiment, the present invention provides extracts, fractions, botanicals derived from Theobroma cacao. a first component selected from the group consisting of citrus aurantium nitrite and mixtures thereof; The present invention relates to a method for producing a plant extract, a plant fraction, a plant chemical component, and a mixture thereof, which is selected from the group consisting of extracts, fractions, phytochemical components, and mixtures thereof derived from the plant extract, A synergistic herbal composition is provided that includes a combination of two components, wherein the concentration of the first component in the composition is: The concentration of the second component varies from 10% to 90% by weight, and the concentration of the second component varies from 90% to 10% by weight. It varies in the range of

[0062] In other exemplary embodiments, the present invention provides extracts, fractions, and the like derived from Theobroma cacao. The first ingredient is selected from the group consisting of phytochemicals, citrus aura, and combinations thereof. Extracts, fractions, phytochemical constituents and mixtures thereof derived from Prunus antifolia are selected from the group consisting of and a second ingredient selected from the group consisting of hydroxybenzoates, ... At least one selected from the group consisting of physiologically or nutritionally acceptable excipients, carriers and diluents. Optionally, the component

[0063] In another embodiment, the present invention provides a method for preventing or controlling obesity and / or overweight. Treat and improve lean body mass and increase browning of white adipose tissue (WAT) / brown adipose tissue Improves the formation of BAT and stabilizes basal metabolic rate (BMR) / energy expenditure, Increases thermogenesis, improves thyroid function, maintains healthy weight, increases satiety, and reduces At least one of the following is selected from the group consisting of supporting body mass, enhancing fat loss, and maintaining lean body mass: Extracts, fractions, and botanicals derived from Theobroma cacao for additional health benefits. a first component selected from biochemical components and combinations thereof; and a Citrus Aurantium and mixtures thereof. and a second ingredient combination that provides a synergistic herbal composition for pharmaceutical or nutraceutical use. or at least one component selected from nutritionally acceptable excipients, carriers and diluents Optionally, pharmaceutically or nutraceutical or nutritionally acceptable excipients, carriers and and diluents are monosaccharides such as glucose, dextrose, fructose, and galactose. ; sucrose, maltose, lactose, lactulose, trehalose, cellobiose, disaccharides, such as, but not limited to, chitobiose; starch and starch glycols; Modified starches such as sodium phosphate, pregelatinized starch, soluble starch, and others Polycarbohydrates such as modified starches; yellow dextrin, white dextrin, maltodextrin Dextrins, such as starch, produced by hydrolysis of glycogen sorbitol, mannitol, inositol, xylitol, isomalt, etc. polyhydric alcohols or sugar alcohols; microcrystalline cellulose, hydroxypropyl cellulose, Examples include, but are not limited to, hydroxypropyl methylcellulose and hydroxyethyl cellulose. No, cellulose-based derivatives; Neusilin, Veegum, Talc, Colloidal Dioxide silicates such as, but not limited to, calcium stearate, stearic acid Metal stearates, such as, but not limited to, magnesium, zinc stearate Organic acids such as citric acid, tartaric acid, malic acid, succinic acid, lactic acid, and L-ascorbic acid; Resolvates, fatty acid esters and esters, acacia, carrageenan, guar gum natural gums, such as, but not limited to, gum, xanthan gum; B vitamins, nicotine Amides, calcium pantothenate, amino acids, casein, gelatin, pectin, agar, etc. Proteins such as, but not limited to; sodium chloride, calcium chloride, diphosphate Organic metal salts such as, but not limited to, calcium, zinc sulfate, and zinc chloride; natural pigments , fragrances, Class I & II preservatives and aqueous solutions of the above listed ingredients, alone or in combination, The solvent is selected from an alcohol solution, a water-alcohol solution, and an organic solution.

[0064] In another embodiment, the present invention provides extracts, fractions, plant extracts, and plant fractions derived from Theobroma cacao. a first component selected from chemical components and combinations thereof; and a Citrus Aurantifida extract. The compounds are selected from extracts, fractions, phytochemical components and combinations thereof derived from Olia. A synergistic herbal composition comprising a combination of second ingredients is provided, wherein the extract or fraction comprises leaves, stems, Tender stems, tender twigs, above-ground parts, whole fruit, fruit peel rind, seeds, flower heads, roots, bark , hardwood or whole plant or mixtures thereof. It is obtained from plant parts.

[0065] In another embodiment, the present invention provides extracts, fractions, plant extracts, and plant fractions derived from Theobroma cacao. a first component selected from chemical components and combinations thereof; and a Citrus Aurantifida extract. The compounds are selected from extracts, fractions, phytochemical components and combinations thereof derived from Olia. A synergistic herbal composition comprising a combination of second ingredients is provided, wherein the extract or fraction is ethanol. C1-C5 alcohols such as ethanol, methanol, n-propanol, and isopropyl alcohol ketones such as acetone and methyl isobutyl ketone, dichloromethane and chloroform Chlorinated solvents such as formaldehyde, water and their mixtures, C1-C7 carbon dioxide such as hexane from the group including hydrogen, esters such as ethyl acetate and the like, and mixtures thereof The solvent is prepared using at least one solvent selected from the group consisting of:

[0066] In another embodiment, the present invention provides extracts, fractions, botanicals derived from Theobroma cacao. a first component selected from the group consisting of citrus aurantium nitrite and mixtures thereof; The present invention relates to a method for producing a plant extract, a plant fraction, a plant chemical component, and a mixture thereof, which is selected from the group consisting of extracts, fractions, phytochemical components, and mixtures thereof derived from the plant extract, The present invention provides a synergistic herbal composition comprising a combination of two ingredients, wherein the extract or fraction comprises at least Towards a biologically active marker compound or extract or fraction of a single phytochemical constituent and standardized as phytochemical marker compounds and groups of phytochemical compounds. The concentration range is 0.1% to 99% by weight of the extract.

[0067] In another embodiment, the present invention provides extracts, fractions, botanicals derived from Theobroma cacao. a first component selected from the group consisting of citrus aurantium nitrite and mixtures thereof; The present invention relates to a method for producing a plant extract, a plant fraction, a plant chemical component, and a mixture thereof, which is selected from the group consisting of extracts, fractions, phytochemical components, and mixtures thereof derived from the plant extract, The present invention provides a synergistic herbal composition comprising a combination of two ingredients, an extract or fraction, and Theobroma capitata. The phytochemical composition of cocoa is standardized to theobromine, and the concentration range of theobromine is , 0.1% to 20% by weight of the composition.

[0068] In another embodiment, the present invention provides extracts, fractions, botanicals derived from Theobroma cacao. a first component selected from the group consisting of citrus aurantium nitrite and mixtures thereof; The present invention relates to a method for producing a plant extract, a plant fraction, a plant chemical component, and a mixture thereof, which is selected from the group consisting of extracts, fractions, phytochemical components, and mixtures thereof derived from the plant extract, A synergistic herbal composition comprising a combination of two ingredients, Citrus aurantifolia The extract or fraction is standardized to limonin, and the concentration range of limonin is It is 0.1% by weight to 10% by weight.

[0069] In another embodiment, the present invention provides extracts, fractions, plant extracts, and plant fractions derived from Theobroma cacao. a first component selected from chemical components and combinations thereof; and a Citrus Aurantifida extract. Extracts, fractions, phytochemical components and mixtures thereof derived from Olia and a second ingredient combination, wherein the composition(s) are in the form of a dry powder. Form, liquid form, beverage, food, dietary supplement or tablet, capsule, soft chew Dosage forms selected from any suitable form, such as crushable or gummy bears It is formulated into

[0070] The composition(s) disclosed above can be used in health foods, i.e. chocolate or nutritional supplements. Solid foods such as nutrition bars, semi-solid foods such as creams, jams or gels, or refrigerated foods Beverages such as Resh drinks and lactic acid bacteria drinks, drops, candy, chewing gum, gummy candy Yogurt, ice cream, pudding, soft red bean jelly, jelly, cookies, tea, Specific health benefits such as soft drinks, juice, milk, coffee, cereal, and snack bars Nutritional / dietary information that can be carefully considered / made into food formulations for health use It can be formulated into a supplement.

[0071] In a further embodiment, the method comprises preventing, controlling, or treating obesity and / or overweight, and reducing fat. Improves fat mass and promotes browning of white adipose tissue (WAT) / brown adipose tissue (BAT) ) formation, increasing the basal metabolic rate (BMR) and stabilizing energy expenditure. , improves thermogenesis, improves thyroid function, maintains a healthy weight, increases satiety, and promotes weight loss and to provide a method for promoting fat loss and maintaining a lean body, Extracts, fractions, phytochemical constituents and their mixtures derived from Theobroma cacao and an extract derived from Citrus aurantifolia. a second component selected from a phytochemical component, a phytochemical component, and a mixture thereof; At least one additional component selected from pharmaceutically acceptable excipients, diluents, and carriers. The present invention also includes supplementing a subject with a suitable dose of a synergistic herbal composition, optionally containing nothing.

[0072] In another embodiment, the present invention provides a method for preventing, controlling, or treating obesity and / or overweight, and for reducing fat. Improves body weight and promotes browning of white adipose tissue (WAT) / brown adipose tissue (BAT) Improves the formation of ATP, increases the basal metabolic rate (BMR) and stabilizes energy expenditure. Increases thermogenesis, improves thyroid function, maintains a healthy weight, increases satiety, and promotes weight loss and to provide a method for supporting and enhancing fat loss and maintaining a lean body, the method comprising: Extracts, fractions, phytochemical constituents and mixtures thereof derived from Theobroma cacao and an extract derived from Citrus aurantifolia, a second component selected from a phytochemical component and a mixture thereof, at least one additional component selected from physiologically acceptable excipients, diluents, and carriers; and optionally supplementing the subject with a suitable dose of a synergistic herbal composition. The obesity treatment involves promoting lipolysis, adipogenesis, inhibiting lipid accumulation, and fibroblast proliferation. Increased fibroblast growth factor-21 (FGF-21), increased uncoupling protein-1 (UCP-1) and β3- This includes at least one of the following: an increase in adrenergic receptors (β3-AR).

[0073] In another embodiment, the present invention provides a method for preventing, controlling, or treating obesity and / or overweight. Improves lean body mass and promotes browning of white adipose tissue (WAT) / brown adipose tissue Improves the formation of BAT (basal metabolic rate) and increases the basal metabolic rate (BMR) / slows energy expenditure stabilizing, improving thermogenesis, improving thyroid function, maintaining a healthy weight, and increasing satiety Helps you lose weight, improves fat loss and maintains lean body mass Extracts, fractions, and other ingredients derived from Theobroma cacao to obtain the health benefits of both A first component selected from phytochemical components and mixtures thereof and Citrus aurantii and mixtures thereof. and a second component selected from pharmaceutically acceptable excipients, diluents, and carriers. The use of a synergistic herbal composition, optionally containing at least one additional ingredient, provide.

[0074] In another embodiment, the present invention provides a method for preventing, controlling, or treating obesity and / or overweight, and for reducing fat. Improves body weight and promotes browning of white adipose tissue (WAT) / brown adipose tissue (BAT) Improves the formation of ATP, increases the basal metabolic rate (BMR) and stabilizes energy expenditure. Increases thermogenesis, improves thyroid function, maintains a healthy weight, increases satiety, and promotes weight loss and to provide a method for supporting and enhancing fat loss and maintaining a lean body, the method comprising: Extracts, fractions, phytochemical constituents and mixtures thereof derived from Theobroma cacao and an extract derived from Citrus aurantifolia, a second component selected from a phytochemical component and a mixture thereof, at least one additional component selected from physiologically acceptable excipients, diluents, and carriers; and optionally supplementing the subject with a suitable dose of a synergistic herbal composition. The administration of a suitable dose to the subject may be in the form of a sustained release tablet or a bolus of the desired therapeutic agent. To achieve these benefits, nanotechnology, microencapsulation, colloidal carrier systems and sustained release polymer-based coatings by other drug delivery systems including I am using it.

[0075] Those skilled in the art may make modifications to the above embodiments without departing from the broad inventive concept thereof. It will be appreciated that the present invention is not limited to the above-described embodiments. The present invention is not limited to the specific embodiments or examples shown, but includes those defined herein. It is intended to cover modifications that fall within the spirit and scope of the invention as defined herein. The examples presented are intended to illustrate the invention but are not to be construed as limiting the invention in any way. It should not be considered as limiting the scope of the invention. [Example]

[0076] Example 1: Preparation of a water extract of Theobroma cacao seeds Theobroma cacao seeds (100g) were finely ground and the powdery raw material was then mixed with the Extraction with water (700 mL) was carried out in a laboratory extractor at room temperature (rt) for 1 hour. The extract was filtered. The crude material used was re-extracted twice with water (2 x 500 mL) under similar conditions. The combined extracts were filtered and concentrated under vacuum to give the residue of the water extract as a powder ( TC-1, 14g).

[0077] Example 2: Ethanol extract and 50% aqueous ethanol extract of Theobroma cacao seeds Preparation of the ingredients Theobroma cacao seeds (100g) were finely ground and the powdery raw material was then mixed with the Extract with 50% aqueous ethanol (700 mL) in a laboratory extractor at room temperature (rt) for 1 hour. The extract was filtered and the raw material used was resuspended in 50% aqueous ethanol under similar conditions twice. The combined extracts were filtered and concentrated under vacuum. The residue of the ethanol extract was obtained (TC-2, 11 g).

[0078] Ethanol extract of Theobroma cacao seeds (100g) (TC-3, 4.4g) was obtained by adapting the same procedure as above using ethanol as the extraction solvent. It was.

[0079] Example 3: Preparation of a 50% aqueous methanol extract of Theobroma cacao seeds Theobroma cacao seeds (100g) were finely ground and the powdery raw material was then mixed with the Extract with 50% aqueous methanol (700 mL) in a laboratory extractor at room temperature (rt) for 1 hour. The extract was filtered and the raw material used was diluted twice under similar conditions with 50% methanol. (2 x 500 mL). The combined extracts were filtered and concentrated under vacuum. The residue of 50% aqueous methanol extract was obtained (TC-4, 10 g).

[0080] Example 4: Preparation of n-butanol extract of Theobroma cacao seeds Theobroma cacao seeds (100g) were finely ground and the powdery raw material was then mixed with the Extraction with n-butanol (700 mL) was carried out in a laboratory extractor at room temperature (rt) for 1 hour. The extract was filtered and the raw material used was resuspended in n-butanol (2 x 50 mL) twice under similar conditions. The combined extracts were filtered and concentrated under vacuum to give n-butadiene. The residue of the knoll extract was obtained (TC-5, 24.5 g).

[0081] Example 5: Standardization of Theobroma Cacao Seed Extract Various extracts of Theobroma cacao seeds were analyzed for theobromine by HPLC. The results are summarized in Table 1.

[0082] [Table 1]

[0083] Example 6: 50% aqueous ethanol & ethanol of Citrus aurantifolia fruit peel Preparation of le extract Finely ground raw material (100g) of dried Citrus aurantifolia fruit peel The powder was extracted with 1:1 ethanol / water ( The extract was filtered and the crude material used was extracted twice under similar conditions. The extract was re-extracted with 1:1 ethanol / water (2 x 500 mL). The combined extracts were filtered. The residue of 50% aqueous ethanol was concentrated under vacuum to give a brown powder (CA-1, 2 1g). Total flavonoids by UV: 9.92%.

[0084] The ethanol extract (CA-2, 9.5 g) was prepared using ethanol as the extraction solvent. was obtained by adapting the procedure described above.

[0085] Similarly, the raw material of dried whole fruit of Citrus aurantifolia (10 0 g) was subjected to the same extraction procedure as above and extracted with 50% aqueous alcohol as a brown powder (15 g). The extract was obtained.

[0086] Example 7: Preparation of aqueous extract of Citrus aurantifolia fruit peel The dried fruit peel of Citrus aurantifolia (100g) was finely ground and powdered. The processed material was extracted with water (700 mL) in a laboratory extractor at room temperature (rt) for 1 h. The extract was filtered and the raw material used was resuspended twice in water (2 x 500 mL) under similar extraction conditions. The combined extracts were filtered and concentrated under vacuum to give a brown powder. The residue of the water extract was obtained (CA-3, 21 g).

[0087] Example 8: Preparation of 50% aqueous methanol extract of Citrus aurantifolia fruit peel Made The dried fruit peel of Citrus aurantifolia (100g) was finely ground and powdered. The processed material was extracted with 50% aqueous methanol (7%) in a laboratory extractor for 1 h at room temperature (rt). The extract was filtered and the raw material used was extracted twice under similar conditions with 500 mL of water. The extract was re-extracted with 0% aqueous methanol (2 x 500 mL). The combined extracts were filtered and Concentration under vacuum gave the residue of the 50% aqueous methanol extract as a brown powder (C. A-4, 30g).

[0088] Example 9: Preparation of butanol extract of Citrus aurantifolia fruit peel The dried fruit peel of Citrus aurantifolia (100g) was finely ground and powdered. The processed material was extracted with n-butanol (700 mL) in a laboratory extractor for 1 h at room temperature (rt). The extract was filtered and the raw material used was extracted twice under similar conditions with n-butanolic acid. The combined extracts were filtered and concentrated under vacuum. The residue of the butanol extract was obtained as a brown powder (CA-5, 7 g).

[0089] Example 10: Standardization of Citrus Aurantifolia Extract Various extracts of Citrus aurantifolia were analyzed by HPLC to determine the amount of limonin. The results are summarized in Table 2.

[0090] [Table 2]

[0091] Example 11: Composition of Theobroma Cacao Water Extract (TC-1) and Citrus Au Preparation of Lantifolia extracts (CA-1 to CA-5) Composition-1(C-1) : Composition-1 was mixed with Theobroma cacao water extract ( TC-1) and Citrus aurantifolia 50% aqueous ethanol extract (CA- 1) was mixed to prepare the product. Composition-2(C-2) : Composition-2 was mixed with Theobroma cacao water extract ( TC-1) and Citrus aurantifolia 50% aqueous ethanol extract (CA- 1) was mixed to prepare the product. Composition-3(C-3) : Composition-3 was mixed with Theobroma cacao water extract ( TC-1) and Citrus aurantifolia 50% aqueous ethanol extract (CA- 1) was mixed to prepare the product. Composition-4(C-4) : Composition-4 was mixed with Theobroma cacao water extract ( TC-1) and Citrus aurantifolia 50% aqueous ethanol extract (CA- 1) was mixed to prepare the product. Composition-5(C-5) : Composition-5 was mixed with Theobroma cacao water extract ( TC-1) and Citrus aurantifolia 50% aqueous ethanol extract (CA- 1) was mixed to prepare the product. Composition-6(C-6) : Composition-6 was mixed with Theobroma cacao water extract ( TC-1) and Citrus aurantifolia ethanol extract (CA-2) were mixed. It was prepared by mixing the above ingredients. Composition-7(C-7) : Composition-7 was mixed with Theobroma cacao water extract ( TC-1) and Citrus aurantifolia ethanol extract (CA-2) were mixed. It was prepared by mixing the above ingredients. Composition-8(C-8) : Composition-8 was mixed with Theobroma cacao water extract ( TC-1) and Citrus aurantifolia ethanol extract (CA-2) were mixed. It was prepared by mixing. Composition-9(C-9) : Composition-9 was mixed with Theobroma cacao water extract ( TC-1) and Citrus aurantifolia water extract (CA-3) were mixed. It was prepared by the following. Composition-10(C-10) : Composition-10 was mixed with Theobroma cacao water extract in a ratio of 1:2. A mixture of Citrus aurantifolia extract (TC-1) and Citrus aurantifolia water extract (CA-3) It was prepared by Composition-11(C-11) : Composition-11 was added to Theobroma cacao water extract in a 1:1 ratio. A mixture of Citrus aurantifolia extract (TC-1) and Citrus aurantifolia water extract (CA-3) It was prepared by Composition-12(C-12) : Composition-12 was mixed with Theobroma cacao water extract in a ratio of 2:1. A mixture of Citrus aurantifolia extract (TC-1) and Citrus aurantifolia water extract (CA-3) It was prepared by Composition-13(C-13) : Composition-13 was added to Theobroma cacao water extract in a ratio of 3:1. A mixture of Citrus aurantifolia extract (TC-1) and Citrus aurantifolia water extract (CA-3) It was prepared by Composition-14(C-14) : Composition-14 was mixed with Theobroma cacao water extract in a ratio of 1:2. (TC-1) and Citrus aurantifolia 50% aqueous methanol extract (C .A-4) was mixed. Composition-15(C-15) : Composition-15 was added to Theobroma cacao water extract in a 1:1 ratio. (TC-1) and Citrus aurantifolia 50% aqueous methanol extract (C .A-4) was mixed. Composition-16(C-16) : Composition-16 was mixed with Theobroma cacao water extract in a ratio of 2:1. (TC-1) and Citrus aurantifolia 50% aqueous methanol extract (C .A-4) was mixed. Composition-17(C-17) : Composition-17 was mixed with Theobroma cacao water extract in a ratio of 1:2. Extract (TC-1) and Citrus Aurantifolia Butanol Extract (CA-5) It was prepared by mixing the above. Composition-18(C-18) : Composition-18 was mixed with Theobroma cacao water extract in a 1:1 ratio. Extract (TC-1) and Citrus Aurantifolia Butanol Extract (CA-5) It was prepared by mixing the above. Composition-19(C-19) : Composition-19 was mixed with Theobroma cacao water extract in a ratio of 2:1. Extract (TC-1) and Citrus Aurantifolia Butanol Extract (CA-5) It was prepared by mixing the above.

[0092] Example 12: Composition of 50% aqueous ethanol extract of Theobroma cacao (TC-2) Preparation of Citrus aurantifolia extracts (CA-1 to CA-5) Composition-20(C-20) : Composition-20, Theobroma cacao 50 in a 1:3 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia 50% aqueous ethanol extract It was prepared by mixing with ethanol extract (CA-1). Composition-21(C-21) : Composition-21, Theobroma cacao 50 in a 1:2 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia 50% aqueous ethanol extract It was prepared by mixing with ethanol extract (CA-1). Composition-22(C-22): Composition-22, Theobroma cacao 50 in a 1:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia 50% aqueous ethanol extract It was prepared by mixing with ethanol extract (CA-1). Composition-23(C-23) : Composition-23, Theobroma cacao 50 in a 2:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia 50% aqueous ethanol extract It was prepared by mixing with ethanol extract (CA-1). Composition-24(C-24) : Composition-24, Theobroma cacao 50 in a 3:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia 50% aqueous ethanol extract It was prepared by mixing with ethanol extract (CA-1). Composition-25(C-25) : Composition-25, Theobroma cacao 50 in a 1:2 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia ethanol extract It was prepared by mixing with the extract (CA-2). Composition-26(C-26) : Composition-26, Theobroma cacao 50 in a 1:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia ethanol extract It was prepared by mixing with the extract (CA-2). Composition-27(C-27) : Composition-27, Theobroma cacao 50 in a 2:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia ethanol extract It was prepared by mixing with the extract (CA-2). Composition-28(C-28) : Composition-28, Theobroma cacao 50 in a ratio of 1:3 % aqueous ethanol extract (TC-2) and Citrus aurantifolia aqueous extract (C .A-3) was mixed. Composition-29(C-29) : Composition-29, Theobroma cacao 50 in a 1:2 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia aqueous extract (C .A-3) was mixed. Composition-30(C-30) : Composition - 30, Theobroma cacao 50 in a 1:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia aqueous extract (C .A-3) was mixed. Composition-31(C-31) : Composition-31, Theobroma cacao 50 in a 2:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia aqueous extract (C .A-3) was mixed. Composition-32(C-32) : Composition-32, Theobroma cacao 50 in a 3:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia aqueous extract (C .A-3) was mixed. Composition-33(C-33) : Composition-33 in a 1:2 ratio with Theobroma cacao 50 % aqueous ethanol extract (TC-2) and Citrus aurantifolia 50% aqueous ethanol extract (TC-2) It was prepared by mixing with ethanol extract (CA-4). Composition-34(C-34) : Composition-34, Theobroma cacao 50 in a 1:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia 50% aqueous ethanol extract (TC-2) It was prepared by mixing with ethanol extract (CA-4). Composition-35(C-35) : Composition-33 in a 2:1 ratio with Theobroma cacao 50 % aqueous ethanol extract (TC-2) and Citrus aurantifolia 50% aqueous ethanol extract (TC-2) It was prepared by mixing with ethanol extract (CA-4). Composition-36(C-36) : Composition-36, Theobroma cacao 50 in a 1:2 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia butanol extract It was prepared by mixing with the extract (CA-5). Composition-37(C-37) : Composition-37, Theobroma cacao 50 in a 1:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia butanol extract It was prepared by mixing with the extract (CA-5). Composition-38(C-38) : Composition-38, Theobroma cacao 50 in a 2:1 ratio % aqueous ethanol extract (TC-2) and Citrus aurantifolia butanol extract It was prepared by mixing with the extract (CA-5).

[0093] Example 13: Theobroma cacao extracts (TC-3, TC-4 & TC-5) and Preparation of Citrus Aurantifolia Extract Compositions (CA-1 to CA-5) Composition-39(C-39) : Composition-39 was mixed with Theobroma cacao ethanol in a ratio of 1:2. Extract of Citrus aurantifolia (TC-3) and 50% aqueous ethanol extract of Citrus aurantifolia It was prepared by mixing with the extract (CA-1). Composition-40(C-40) : Composition-40 in a 1:1 ratio with Theobroma cacao ethanol Extract of Citrus aurantifolia (TC-3) and 50% aqueous ethanol extract of Citrus aurantifolia It was prepared by mixing with the extract (CA-1). Composition-41(C-41) : Composition-41 was mixed with Theobroma cacao ethanol in a 2:1 ratio. Extract of Citrus aurantifolia (TC-3) and 50% aqueous ethanol extract of Citrus aurantifolia It was prepared by mixing with the extract (CA-1). Composition-42(C-42) : Composition-42 in a 1:2 ratio with Theobroma cacao ethanol Nol extract (TC-3) and Citrus aurantifolia ethanol extract (C. A-2) was mixed. Composition-43(C-43) : Composition-43 was added to Theobroma cacao ethanol in a 1:1 ratio. Nol extract (TC-3) and Citrus aurantifolia ethanol extract (C. A-2) was mixed. Composition-44 (C-44) : Composition-44 was mixed with Theobroma cacao ethanol in a 2:1 ratio. Nol extract (TC-3) and Citrus aurantifolia ethanol extract (C. A-2) was mixed. Composition-45(C-45) : Composition-45 in a 1:2 ratio with Theobroma cacao ethanol Nol extract (TC-3) and Citrus aurantifolia water extract (CA-3) It was prepared by mixing the above. Composition-46(C-46) : Composition-46 in a 1:1 ratio with Theobroma cacao ethanol Nol extract (TC-3) and Citrus aurantifolia water extract (CA-3) It was prepared by mixing the above. Composition-47(C-47) : Composition-47 was mixed with Theobroma cacao ethanol in a 2:1 ratio. Nol extract (TC-3) and Citrus aurantifolia water extract (CA-3) It was prepared by mixing the above. Composition-48(C-48) : Composition-48 is a Theobroma cacao ethanol extract ( TC-3) and Citrus aurantifolia 50% aqueous methanol extract (CA- 4) in a 1:2 ratio. Composition-49(C-49) : Composition-49 in a 1:1 ratio with Theobroma cacao ethanol Nol extract (TC-3) and Citrus aurantifolia 50% aqueous methanol extract It was prepared by mixing with the extract (CA-4). Composition-50(C-50) : Composition-50, Theobroma cacao ethanol in a 2:1 ratio Nol extract (TC-3) and Citrus aurantifolia 50% aqueous methanol extract It was prepared by mixing with the extract (CA-4). Composition-51(C-51) : Composition-51 in a 2:1 ratio with Theobroma cacao ethanol butanol extract (TC-3) and Citrus aurantifolia butanol extract (C. A-5) was mixed. Composition-52(C-52) : Composition-52 in a 1:1 ratio with Theobroma cacao ethanol butanol extract (TC-3) and Citrus aurantifolia butanol extract (C. A-5) was mixed. Composition-53(C-53) : Composition-53 was mixed with Theobroma cacao ethanol in a 2:1 ratio. butanol extract (TC-3) and Citrus aurantifolia butanol extract (C. A-5) was mixed. Composition-54(C-54) : Composition-54, Theobroma cacao 50 in a 1:2 ratio % methanol extract (TC-4) and Citrus aurantifolia water extract (CA -3) was mixed. Composition-55(C-55) : Composition-55, Theobroma cacao 50 in a 1:1 ratio % methanol extract (TC-4) and Citrus aurantifolia water extract (CA -3) was mixed. Composition-56(C-56) : Composition-56, Theobroma cacao 50 in a 2:1 ratio % methanol extract (TC-4) and Citrus aurantifolia water extract (CA -3) was mixed. Composition-57(C-57) : Composition-57, Theobroma cacao 50 in a 1:2 ratio % methanol extract (TC-4) and Citrus aurantifolia 50% ethanol It was prepared by mixing with the extract (CA-1). Composition-58(C-58) : Composition-58, Theobroma cacao 50 in a 1:1 ratio % methanol extract (TC-4) and Citrus aurantifolia 50% ethanol It was prepared by mixing with the extract (CA-1). Composition-59(C-59) : Composition-59, Theobroma cacao 50 in a 2:1 ratio % methanol extract (TC-4) and Citrus aurantifolia 50% ethanol It was prepared by mixing with the extract (CA-1). Composition-60(C-60) : Composition-60 in a 1:2 ratio with Theobroma cacao Nol extract (TC-5) and Citrus aurantifolia water extract (CA-3) It was prepared by mixing the above. Composition-61(C-61) : Composition-61 in a 1:1 ratio with Theobroma cacao Nol extract (TC-5) and Citrus aurantifolia water extract (CA-3) It was prepared by mixing the above. Composition-62(C-62) : Composition-62 in a 2:1 ratio with Theobroma cacao Nol extract (TC-5) and Citrus aurantifolia water extract (CA-3) It was prepared by mixing the above. Composition-63(C-63): Composition-63 was mixed with Theobroma cacao in a 1:2 ratio. Nol extract (TC-5) and Citrus aurantifolia 50% ethanol extract (CA-1) was mixed with the above. Composition-64(C-64) : Composition-64 in a 1:1 ratio with Theobroma cacao Nol extract (TC-5) and Citrus aurantifolia 50% ethanol extract (CA-1) was mixed with the above. Composition-65(C-65) : Composition-65 in a 2:1 ratio with Theobroma cacao Nol extract (TC-5) and Citrus aurantifolia 50% ethanol extract (CA-1) was mixed with the above.

[0094] Example 14: Composition Formulation Composition-66(C-66) : Composition-66 contains 60 g of theobroma cacao water extract ( TC-1), 30g of Citrus aurantifolia 50% aqueous ethanol extract ( CA-1), prepared by mixing 9g of maltodextrin and 1g of syloid. Ta. Composition-67(C-67) : Composition-67, in the presence of ethanol / water, 53.33 g of Theobroma cacao water extract (TC-1), 26.67 g of Citrus aurante 50% aqueous ethanol extract of Ipomoea aquatica (CA-1), 18g of maltodextrin and and 2 g of syloid were mixed and dried to obtain a composition. Composition-68(C-68) Composition-68 is prepared by dissolving 53.2 g of ethanol / water in the presence of Theobroma cacao water extract (TC-1), 26.6g of Citrus Aurantifolia 50% aqueous ethanol extract of Oria (CA-1), 18.2g of Glusidex-12 D and 2 g of syloid were mixed and dried to obtain a composition.

[0095] Example 15: General procedure for adipogenesis inhibition assay Mouse 3T3-L1 preadipocytes (50,000 cells / well in 500 μL) were cultured at 48°C. Cells were seeded in well-culture plates and incubated at 37°C in a humidified atmosphere of 5% CO2 and 10% DMEM (Dulbecco's Modified Eagle's Medium) containing FBS The cells were maintained in a medium containing 100% soluble cellulose. The cells were allowed to reach confluence (~2 days). After that, the cells were soaked in 0.5 mM 3-isobutyl-1-methylpropional in DMEM with 10% FBS. Contains Santin (IBMX), 1 μM dexamethasone, and 500 nM insulin The cells were treated with different concentrations of test samples for 48 hours in differentiation medium (DM) containing The cell culture medium was diluted with 100% DMEM containing 10% FBS and the relative concentrations of the test samples. nM insulin, along with the test samples of the relative concentrations Post-DM was replaced with new ones every 48 hours until day 6. Eight days after the start of differentiation, cells were subjected to Oil Red O staining.

[0096] Oil Red O staining: Aspirate the medium and add 0.5 mL of 10% formaldehyde to each well. and allowed to incubate at room temperature for 2 hours. The formaldehyde was removed from the wells and 0.25 mL of 60% isopropanol was added and the plate was allowed to dry completely. A microliter of red oil O was added and allowed to incubate in the dark at room temperature for 20 minutes. The red oil was aspirated and the plates were washed four times with distilled water. The plates were allowed to dry completely and then incubated for 15 min. Add 0 μL of 100% isopropanol, mix thoroughly and dispense 75 μL into a 96-well plate. Transfer to an assay plate and measure at 550 nm using a Spectramax 5e plate reader. The absorbance was measured at 1000 nm. The percentage of inhibition of adipogenesis was calculated using the following formula:

[0097]

number

[0098] Using the above procedure, all compositions were screened for their inhibition of adipogenesis. The results are shown in Tables 3, 4 & 5.

[0099] Table 3: Theobroma cacao extract (TC-1) and Citrus aurantifolia Adipogenic activity of compositions containing extracts (CA-1 to CA-5) [Table 3]

[0100] Table 4: Theobroma cacao extract (TC-2) and Citrus aurantifolia Adipogenic activity of compositions containing extracts (CA-1 to CA-4) [Table 4]

[0101] Table 5: Theobroma cacao extracts (TC-3, TC-4, TC-5) and Citra A composition containing S. aurantifolia extract (CA-1 to CA-5) adipogenic activity [Table 5]

[0102] Example 16: General procedure for augmentation of adipolysis assays Mouse 3T3-L1 preadipocytes (50,000 cells / well in 500 μL) were cultured at 48°C. Cells were seeded in well-culture plates and incubated at 37°C in a humidified atmosphere of 5% CO2 and 10% Cells were maintained in DMEM containing FBS until confluent (~2 days). Then, the cells were incubated in DMEM with 10% FBS, 0.5 nM IBMX, 1 μM dextran, and 1 μM PEG-400. The cells were cultured for 48 h using differentiation medium (DM) containing methasone and 500 mM insulin. The cell culture medium was 100 ml of DMEM containing 10% FBS. The post-DM was changed to one containing 48 nM insulin and maintained for 48 h until day 6. On day 7, the medium was aspirated from the wells and phenol red was removed. The cells were washed with 600 μL of DMEM containing no BSA. Test samples of different differentiation concentrations were prepared in phenol red-free DMEM medium. The samples were treated with 50 μL of ethanol and incubated at 37°C for 4 hours in a CO2 incubator. After incubation, 25 μL of cell-free supernatant was removed from the wells. The cells were then transferred to a 96-well assay plate and processed for the glycerol assay.

[0103] Glycerol Assay: 25 microliters of standard or sample was added to a 96-well plate. In a 100 μL assay plate, add 100 μL of glycerol reagent [ATP: 20.65 mg, M gCl2: 46.20 mg, N-ethyl-N-(3-sulfopropyl)-m-anisidine -Sodium salt: 31.10 mg, Aminoantipyro l): 1.90 mg, glycerol kinase: 10.2 μL, glycerol oxidase 125 μL of ATP and 62.5 μL of HRP were dissolved in 50 mL of 1x PBS. The plate was incubated at room temperature for 15 minutes. The absorbance was measured on a Spectramax5e plate. The measurement was performed at 550 nm using a reader. A seven-point glycerol standard curve was performed from 0.781 to 0.785. The increase in lipolysis was estimated using the following formula: did.

[0104]

number

[0105] Using the above procedure, all compositions were screened for their increased lipolysis. The results are shown in Tables 6 and 7.

[0106] Table 6: Theobroma cacao extract (TC-1) and Citrus aurantifolia Adipolytic activity of compositions containing extracts (CA-1 to CA-4) [Table 6]

[0107] Table 7: Theobroma cacao extracts (TC-2 and TC-3) and Citrus au Lait extract Adipose tissue containing Lantifolia extract (CA-1 to CA-5) cis-active [Table 7]

[0108] Example 17: General procedure for FGF21 assay Mouse 3T3-L1 preadipocytes (150,000 cells / well in 3 mL) were cultured in a 6-well plate. The cells were seeded onto cell culture plates and incubated at 37°C in a humidified atmosphere of 5% CO2 with 10% FB. The cells were maintained in DMEM medium containing 0.5% ethanol and 4.5 g / L glucose. After allowing the cells to become confluent (~2 days), they were cultured in DMEM with 10% FBS for 0.5 min. It contained 1 mM IBMX, 1 μM dexamethasone, and 500 nM insulin. The differentiation was induced for 48 hours using differentiation medium (DM) containing 10% F Change to post-DM containing 100 nM insulin in DMEM with BS. The post-DM was replaced with new ones every 48 hours until day 6. On day 7, the medium was removed from the wells. Aspirate the supernatant and use DMEM medium containing 1% FBS without phenol red. The cells were then resuspended in DMEM medium containing 1% FBS (total volume of 1 mL). The cells were treated with test samples (50 μL) of different differentiation concentrations at 4°C, and then incubated at 4°C for 1 hour at CO The mixture was further incubated at 37°C for 48 hours. Cell-free supernatant was collected from the wells and added for FGF21 analysis by ELISA. FGF21 ELISA (R&D Systems Cat#DY2539) , was performed according to the manufacturer's protocol.

[0109] The rate of increase in FGF21 was calculated using the following formula:

[0110]

number

[0111] Using the above procedure, the selected composition is evaluated for its FGF-21 increase over the control. The results of the screening are shown in Table 8.

[0112] Table 8: Formulas containing Theobroma cacao extract and Citrus aurantifolia extract FGF21 activity of the composition [Table 8]

[0113] Example 18: General procedure for UCP-1 assay Mouse 3T3-L1 preadipocytes (150,000 cells / well in 3 mL) were cultured in a 6-well plate. The cells were seeded onto cell culture plates and incubated at 37°C in a humidified atmosphere of 5% CO2 with 10% FB. The cells were maintained in DMEM medium containing 0.5% ethanol and 4.5 g / L glucose. After allowing the cells to become confluent (~2 days), they were cultured in DMEM with 10% FBS for 0.5 min. It contained 1 mM IBMX, 1 μM dexamethasone, and 500 nM insulin. The differentiation was induced for 48 hours using differentiation medium (DM) containing 10% F Change to post-DM containing 100 nM insulin in DMEM with BS. The post-DM was replaced with new ones every 48 hours until day 6. On day 7, the medium was removed from the wells. Aspirate the supernatant and use DMEM medium containing 1% FBS without phenol red. The cells were then resuspended in DMEM medium containing 1% FBS (total volume of 1 mL). The cells were treated with test samples (50 μL) of different differentiation concentrations at 4°C, and then incubated at 4°C for 1 hour at CO The plates were incubated at 37°C for a further 48 hours.

[0114] Western Blot: After incubation, place the cell culture plate on an ice tray and rinse twice with 1x PBS. Washed. Add 80 microliters of lysis buffer (10 mM Tris-HCl pH 7. 4, 150 mM NaCl, 1 mM EDTA, 1 mM PMSF, 10 μg / mL Protinin, 10 μg / mL leupeptin, 1% Triton X-100, 1 mM Na F, 1 mM Na3VO4, 0.5% sodium deoxycholate, and 1 μM Phenol statin) was added to each well, and the cell lysates were collected in microfuge tubes. The microfuge tube was sonicated for 1 minute. Cellular proteins were extracted at 21952 x g. After centrifugation, the cells were collected and protein was determined using the Pierce BCA protein assay. Quantification using a kit (Thermo Scientific Cat# 23225) Protein samples were subjected to SDS-polyacrylamide gel electrophoresis. The separated proteins were analyzed by wet blotting. Briefly, 10 μg of protein was transferred onto a nitrocellulose membrane. The mixture was loaded onto a gel (10% resolution) and electrophoresed at approximately 100 V for 1 hour and 40 minutes. At the end, the transfer system was placed in a 4°C chamber (100 V for 2 hours) to allow for nitrocellulose transfer. After transfer, UCP-1 was incubated for 18 hours at 4°C. Anti-UCP-1 antibody (Thermo Scientific Cat#PA1-2 β-actin was probed with 4894 (1:10000 dilution). Incubated anti-β-actin antibody (Sigma Cat# A4700-100 The antibodies were probed with a peroxidase-conjugated mouse anti-goat secondary antibody (1:10000 dilution). Antibody (Jackson Immuno Research Cat#205-035-1 08, diluted 1:10000) was added and allowed to incubate at room temperature for 30 minutes. Fluorescence was detected using a chemiluminescent substrate (Thermo Scientific Cat#34080). The lot was developed and analyzed using a Bio-Rad molecular imaging system (model: ChemiDOC XRS+). Images were captured using a Fluorescence Intensity Analyzer (FIND) and a Fluorescence Imaging System (FII). The intensity of the UCP-1 protein band was measured using a Fluorescence Intensity Analyzer (FIND) and a Fluorescence Imaging System (FII). Relative indices were calculated using eam MI software and normalized using β-actin. The percentage of UCP-1 expression over control was calculated using the following formula:

[0115]

number

[0116] Using the above procedure, the selected composition is compared for its % UCP-1 expression over the control. The results of the screening are shown in Table 9.

[0117] Table 9: Composition of Theobroma cacao extract and Citrus aurantifolia extract UCP-1 expression [Table 9]

[0118] Example 19: General procedure for β3AR assay Mouse 3T3-L1 preadipocytes (150,000 cells / well in 3 mL) were cultured in a 6-well plate. The cells were seeded onto cell culture plates and incubated at 37°C in a humidified atmosphere of 5% CO2 with 10% FB. The cells were maintained in DMEM medium containing 0.5% ethanol and 4.5 g / L glucose. After allowing the cells to become confluent (~2 days), they were cultured in DMEM with 10% FBS for 0.5 min. It contained 1 mM IBMX, 1 μM dexamethasone, and 500 nM insulin. The differentiation was induced for 48 hours using differentiation medium (DM) containing 10% F Change to post-DM containing 100 nM insulin in DMEM with BS. The post-DM was replaced with new ones every 48 hours until day 6. On day 7, the medium was removed from the wells. Aspirate the supernatant and use DMEM medium containing 1% FBS without phenol red. The cells were then resuspended in DMEM medium containing 1% FBS (total volume of 1 mL). The cells were treated with test samples (50 μL) of different differentiation concentrations at 4°C, and then incubated at 4°C for 1 hour at CO The plates were incubated at 37°C for a further 48 hours.

[0119] Western Blot: After incubation, place the cell culture plate on an ice tray and rinse twice with 1x PBS. Washed. Add 80 microliters of lysis buffer (10 mM Tris-HCl pH 7. 4, 150 mM NaCl, 1 mM EDTA, 1 mM PMSF, 10 μg / mL Protinin, 10 μg / mL leupeptin, 1% Triton X-100, 1 mM Na F, 1 mM Na3VO4, 0.5% sodium deoxycholate, and 1 μM Phenol statin) was added to each well, and the cell lysates were collected in microfuge tubes. The microfuge tube was sonicated for 1 minute. Cellular proteins were extracted at 21952 x g. After centrifugation, the cells were collected and protein was determined using the Pierce BCA protein assay. Quantification was performed using a kit (Thermo Scientific Cat#23225). Protein samples were subjected to SDS-polyacrylamide gel electrophoresis and analyzed. The separated proteins were then subjected to wet blotting. Briefly, 10 μg of protein was transferred onto a cellulose membrane. The gel was placed on a 10% decomposition column and electrophoresed at approximately 100 V for 1 hour and 40 minutes. Instead, the transfer system was placed in a 4°C chamber (100V for 2 hours) to generate nitrocells. After transfer, β3AR was incubated at 4°C for 18 hours. Anti-β3AR antibody (Biorbyt Cat# orb221343, 1:500 dilution) β-actin was probed with anti-β-actin antibody, which was incubated at room temperature for 2 hours. Cutin antibody (Sigma Cat#A4700-100uL, 1:10000 dilution) was used. A peroxidase-conjugated mouse anti-goat secondary antibody (Jackson Immunosorbent Assay) was used. No Research Cat#205-035-108, 1:10000 dilution) The mixture was then added to the plate and incubated at room temperature for 30 minutes. Finally, a chemiluminescent substrate (Thermo S The blots were developed using Bio-Rad (Bio-Rad Scientific Cat#34080). Images were captured using a molecular imaging device (model: ChemiDOC XRS+). The intensity of the β3AR protein band was analyzed using Carestream MI software. The relative index was calculated and normalized using β-actin. was calculated using the following formula:

[0120]

number

[0121] Using the above procedure, all compositions were screened for β3AR expression above their control. The results are shown in Table 10.

[0122] Table 10: β3AR composition of Theobroma cacao and Citrus aurantifolia Expression [Table 10]

[0123] Example 20: Theobroma in a high-fat diet-induced obesity model in Sprague-Dawley rats Cacao water extract (TC-1) and Citrus aurantifolia 50% aqueous ethanol Composition-6 containing Nol extract (CA-1), maltodextrin and syloid In vivo anti-obesity activity of 7

[0124] Induction: Selected healthy Sprague-Dawley rats were treated with either normal controls (N=7) or high-fat The subjects were randomly assigned to the high-fat diet-induced obesity group (n=34). All animals were fed a high-fat diet to prevent the dietary intervention during the first 4-week induction period. At the end of the induction period, animals were classified as overweight based on body weight. They were randomized into four groups (n=7): G2 - overweight controls; G3 - obese controls; -Composition-67(100mg / Kg, po), G4-Composition-67(300mg / K g, po) and G5-sibutramine (10 mg / Kg, po).

[0125] Treatment: A 4-week induction phase followed by 4 weeks of treatment with the assigned test substance or vehicle. The animals were treated orally (using oral gavage). 100 mg (G3) or 300 mg / kg ( G4) Supplement with Composition-67 (C-67) or sibutramine at 10 mg / kg body weight Animals in the control group received vehicle (10 mL of 0.5% CMC in water) during this period. During the treatment phase, all animals received standard rodent chow until the end of the study. was supplied.

[0126] Parameters assessed: Body weights of individual animals were recorded weekly for the entire duration of the study. The average body weight was measured. During the treatment phase and at the end of treatment, the weight gain was compared with the initial weight, respectively. The gain was calculated weekly, and the results are shown in Figures 1A and 1B.

[0127] Dietary calorie consumption: Record the total amount of food consumed by the experimental rats during the treatment phase of the study. The total amount of feed consumed over the 28 days was averaged to estimate daily feed consumption (in grams). The average feed consumption (grams) was 3.43KJ / gram for normal solid feed and 3.43KJ / gram for high-fat feed. The amount of energy (KJ / gram) provided by the high-fat diet (5.15 KJ / gram) The average daily dietary calorie consumption (KJ / day) of the experimental group was , as shown in (Figure 2).

[0128] Subsequently, animals were euthanized and visceral (retroperitoneal, epididymal, perirenal and mesenteric) adipose tissue was removed from the rats. The samples were taken from the tank and weighed using an electronic balance with a sensitivity of 0.01 g (Mettler To (Ledo, Columbus, OH). Observations regarding changes in visceral fat weight are shown in Figure 1. It is summarized in 3.

[0129] Morphometry of adipose tissue: Epididymal adipose tissue collected from individual animals was analyzed by 10% morphometry. Paraffin-embedded tissue was cut into 5 μm sections and the sections were analyzed using the following standard procedures. Hematoxylin-eosin stained tissue sections were microscopically analyzed. Observation was performed at 20x magnification (Axio Scope A1, Carl Zeiss GmbH, Jena, Germany). The pixel size of fat cells was estimated using 0. The area of ​​each cell was estimated using a converted scale of 1694 μm. is shown in Figure 4.

[0130] Serum biomarker assessment: At the end of the study, blood samples were taken from all animals. Serum was separated from the liquid sample and analyzed using a commercially available ELISA kit (e.g., leptin ELISA kit, EMD Millipore, Billerica, MA; T3 and T4 ELISA kits Using a lab kit (Calbiotech, EL Cajon, CA), leptin, triiodothyronine, and thiamin were measured. Dothyronine (T3) and thyroxine (T4) were analyzed. The assay was performed according to the instructions provided. Serum leptin data are summarized in Figure 5. Thyroid hormone balance (T3 / T4) , were calculated for all animals and the results are summarized in Figure 6 .

[0131] Adipocyte immunohistochemistry: UCP-1 immunohistochemistry was performed using DAB staining according to the kit instructions. The color method was performed on paraffin-embedded epididymal adipose tissue (EMD Millipore , Billerica, MA). Tissue classification using UCP-1 antibody (Invitrogen, C Arlsbad, CA), followed by reaction with streptavidin-conjugated HRP. Finally, diaminobenzedene (DAB) was used. The antibody-specific color reaction was developed using the Axio Obs Server Z1 microscope (Carl-Zeiss, Oberkochen, Germany) Representative micrographs of UCP-1 stained epididymal adipose tissue sections captured using a microscope. The truth is shown in Figure 7.

Claims

1. 1. An herbal composition for obtaining at least one health benefit selected from: (A) preventing, controlling, or treating obesity and / or overweight; (B) improving lean body mass; (C) enhancing browning of white adipose tissue (WAT) and / or enhancing brown adipose tissue (BAT) formation; (D) increasing basal metabolic rate (BMR) and / or stabilizing energy expenditure; (E) enhancing thermogenesis; (F) improving thyroid function; (G) maintaining a healthy body weight; (H) increasing satiety; (I) assisting in weight loss; (J) enhancing fat loss; and (K) maintaining lean body mass, a first component selected from extracts, fractions, phytochemical components and mixtures thereof derived from Theobroma cacao seeds, wherein said extracts, fractions, phytochemical components and mixtures thereof are produced by extraction with at least one solvent selected from water, C1-C4 alcohols, or mixtures of water and C1-C4 alcohols; and a second component selected from extracts, fractions, phytochemical components and mixtures thereof derived from the peel of Citrus aurantifolia fruit, wherein said extracts, fractions, phytochemical components and mixtures thereof are produced by extraction with at least one solvent selected from water, C1-C4 alcohols, or mixtures of water and C1-C4 alcohols; A herbal composition in which the weight of Theobroma cacao seed derived components varies from 10% to 90% and the weight of Citrus aurantifolia fruit peel derived components varies from 90% to 10%.

2. 10. The herbal composition of claim 1, wherein the Theobroma cacao seed extract or fraction is standardized to theobromine, the theobromine concentration ranging from 0.1% to 20% by weight of the composition.

3. 2. The herbal composition of claim 1, wherein the Citrus aurantifolia fruit peel extract or fraction is standardized to limonin, and the concentration of limonin ranges from 0.1% to 10% by weight of the composition.

4. 10. The herbal composition of claim 1, optionally comprising at least one ingredient selected from a pharmaceutically, nutraceutical or nutritionally acceptable excipient, carrier or diluent.

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