Patent Publication Number: US-2010113584-A1

Title: Novel use of (-)-epigallocatechin gallate

Description:
The present invention relates to novel uses of (−)-epigallocatechin gallate (EGCG) and dietary supplements, nutraceutical compositions, food, feed and beverages containing (−)-epigallocatechin gallate. 
     The compositions of the present invention are particularly intended for increasing the desire, willingness and/or motivation of animals (including humans), especially mammals, to exercise, to perform and/or to run, especially to perform endurance exercise and furthermore to accelerate training success. 
     The term “EGCG” as used herein denotes (−)-epigallocatechin gallate and/or one or more of its derivatives (e.g. esterified forms, glycosides, sulphates) thereof. Especially preferred is (−)-epigallocatechin gallate itself. 
     The terms “(−)-epigallocatechin gallate” and “EGCG” as used herein encompass naturally occurring sources (one or more components) containing EGCG, for example green tea and/or green tea extracts, e.g. aqueous green tea extracts containing EGCG and the like. According to the present invention it may also be advantageous to use synthetically manufactured EGCG. 
     In preferred embodiments of the present invention the used EGCG has a high purity, e.g. a purity of at least 80%, preferably of at least 85%, more preferably of at least 90%, even more preferably of at least 92%, most preferably of at least 94%. Especially preferred is also an aqueous green tea extract containing EGCG in an amount of at least 80% (preferred of at least 85%, more preferred of at least 90%, even more preferred of at least 92%, most preferred of at least 94%), based on the total amount of the extract. 
     The term “nutraceutical” as used herein denotes usefulness in both, the nutritional and pharmaceutical field of application. Thus, “nutraceutical compositions” according to the present invention can serve as supplements to food, feed and beverages, dietary supplements and as pharmaceutical formulations which may be solid—such as capsules or tablets—or liquid—such as solutions or suspensions. It is evident from the foregoing that the term “nutraceutical composition” also comprises food, feed and beverages containing EGCG. 
     In particular the present invention refers to the use of EGCG to increase
         the desire to exercise;   the willingness to run and/or perform;   the motivation to move and/or   explorative behavior
 
of animals, preferably of mammals.
       

     Furthermore the present invention refers to the use of EGCG for
         accelerating training success;   leading to a faster training success;
 
of animals, preferably of mammals.
       

     The terms “desire to exercise”, “willingness to run and/or perform” and “motivation to move” as used herein include motivation for exercise, self-motivation (for exercise), willingness to perform endurance exercise as well as the urge to move. 
     According to the present invention the animals are preferably mammals, more preferably mammals selected from the group consisting of humans and pets (especially horses, dogs, cats and/or small animals, e.g. hamsters and/or guinea pigs). Most preferred are humans. 
     The EGCG—preferably highly purified EGCG as defined above—may be administered in form of (fortified) food or (fortified) feed, dietary supplements, beverages, tablets, granules, capsules, pastes, gels, powders, food additives, feed additives, or effervescent formulations. Further examples are cereals, cereal bars, dairy products (e.g. milk, buttermilk, soured milk, yogurt (drinks), curd, quark desserts and so on) and pasta (especially tomato) sauces containing EGCG. 
     In a preferred embodiment of the present invention EGCG is administered on a regular (preferred daily) basis and/or in due time (e.g. several weeks) before the beginning of the exercise or before a competition, match, tournament and the like. 
     In preferred embodiments of the uses as given above the daily dosage of EGCG varies from 0.1 to 30 mg per kg body weight per day, preferably from 1.4 to 8.5 mg per kg body weight per day, more preferably from 2 to 4.5 mg per kg body weight per day, most preferably from 4.0 to 4.5 mg per kg body weight per day. 
     Thus, for humans with a weight of 70 kg, the daily dosage of EGCG varies from 7 to 2100 mg, preferably from 100 to 600 mg, more preferably from 140 to 300 mg; most preferably it is about 300 mg per day. 
     Skeletal muscles contain two main types of fibers, which differ in the mechanism they use to produce ATP; the amount of each type of fiber varies from muscle to muscle and from person to person. 
     Red (“slow-twitch”) fibers have more mitochondria, store oxygen in myoglobin, rely on aerobic metabolism, have a greater capillary to volume ratio and are associated with endurance; these produce ATP more slowly. Marathon runners tend to have more red fibers, generally through a combination of genetics and training. “Slow-twitch” fibers are termed Type I fibers (see Table below). Therefore, the more Type I fibers an individual has the better its skeletal muscle can conduct endurance activities and the higher is its resistance to fatigue. 
     White (“fast-twitch”) fibers have fewer mitochondria, are capable of more powerful (but shorter) contractions, metabolize ATP more quickly, have a lower capillary to volume ratio, and are more likely to accumulate lactic acid. Weightlifters and sprinters tend to have more white fibers. Three major fiber types are defined as “fast-twitch” fibers, which are Type IIa fibers, Type IId/x fibers and Type IIb fibers (Table). Among these fibers Type IIa fibers have a predominantly aerobic metabolism and are therefore utilized to support endurance activities. Type IIb fibers on the other hand have a predominantly glycolytic metabolism, are fatiguing quickly and are not suitable for long-term endurance activities. Therefore, if an individual has more Type IIa fibers its skeletal muscle has a higher capacity to conduct endurance activities whereas individuals with more abundant Type IIb fibers are likely to fatigue quicker during endurance activities. 
     
       
         
           
               
             
               
                 TABLE 
               
             
            
               
                   
               
               
                 Characteristics of muscle fiber types 
               
            
           
           
               
               
               
               
               
            
               
                 Fiber Type 
                 Type I 
                 Type IIa 
                 Type IId/x 
                 Type IIb 
               
               
                   
               
               
                 Contraction time 
                 Slow 
                 Moderately 
                 Fast 
                 Very fast 
               
               
                   
                   
                 fast 
               
               
                 Size of motor neuron 
                 Small 
                 Medium 
                 Large 
                 Very large 
               
               
                 Resistance to fatigue 
                 High 
                 Fairly high 
                 Intermediate 
                 Low 
               
               
                 Activity Used for 
                 Aerobic 
                 Long-term 
                 Short-term 
                 Short-term 
               
               
                   
                   
                 aerobic 
                 anaerobic 
                 anaerobic 
               
               
                 Force production 
                 Low 
                 Medium 
                 High 
                 Very high 
               
               
                 Mitochondrial density 
                 High 
                 High 
                 Medium 
                 Low 
               
               
                 Capillary density 
                 High 
                 Intermediate 
                 Low 
                 Low 
               
               
                 Oxidative capacity 
                 High 
                 High 
                 Intermediate 
                 Low 
               
               
                 Glycolytic capacity 
                 Low 
                 High 
                 High 
                 High 
               
               
                 Major storage fuel 
                 Triglyc- 
                 Creatine 
                 Creatine 
                 Creatine 
               
               
                   
                 erides 
                 phosphate, 
                 phosphate, 
                 phosphate, 
               
               
                   
                   
                 glycogen 
                 glycogen 
                 glycogen 
               
               
                   
               
            
           
         
       
     
     Accordingly a further object of the present invention is the use of EGCG to increase
         the number of oxidative (“slow-twitch”) muscle fibers;   the number of aerobic skeletal muscle fibers which can help to perform endurance exercise and/or   fiber type shift towards aerobic skeletal muscle fibers
 
in animals.
       

     According to the present invention the animals are preferably mammals, more preferably mammals selected from the group consisting of humans and pets (especially horses, dogs, cats and/or small animals, e.g. hamsters and/or guinea pigs). Most preferred are humans. 
     It was not to be forseen by the person skilled in the art that the consumption of EGCG would exert changes in fiber type composition of skeletal muscle similar to regular endurance exercise, even without any (endurance) exercise performed. It was especially surprising that the number of oxidative muscle fibers was increased independent from any physical exercise, i.e. in sedentary and trained individuals similarly. 
     The preferences mentioned above for the purity of EGCG, the forms in which it can be administered, and its daily dosage also apply here. 
     Further object of the present invention is a method for increasing
         the desire to exercise;   the willingness to run and/or perform;   the motivation to move and/or   explorative behavior
 
of animals, preferably of mammals, said method comprising the step of administering an effective dose of EGCG to said animal, whereas the effective dose of EGCG varies from 0.1 to 30 mg per kg body weight per day, preferably from 1.4 to 8.5 mg per kg body weight per day, more preferably from 2 to 4.5 mg per kg body weight per day, most preferably from 4.0 to 4.5 mg per kg body weight per day.
       

     Further object of the present invention is a method for increasing
         the number of oxidative (“slow-twitch”) muscle fibers;   the number of aerobic skeletal muscle fibers which can help to perform endurance exercise and/or   fiber type shift towards aerobic skeletal muscle fibers
 
in animals, preferably of mammals, said method comprising the step of administering an effective dose of EGCG to said animal, whereas the effective dose of EGCG varies from 0.1 to 30 mg per kg body weight per day, preferably from 1.4 to 8.5 mg per kg body weight per day, more preferably from 2 to 4.5 mg per kg body weight per day, most preferably from 4.0 to 4.5 mg per kg body weight per day.
       

     Further object of the present invention is a method for
         accelerating training success;   leading to a faster training success;
 
of animals, preferably of mammals, said method comprising the step of administering an effective dose of EGCG to said mammal, whereas the effective dose of EGCG varies from 0.14 to 30 mg per kg body weight per day, preferably from 1.4 to 8.5 mg per kg body weight per day, more preferably from 2 to 4.5 mg per kg body weight per day, most preferably from 4.0 to 4.5 mg per kg body weight per day.
       

     The preferences and explanations mentioned above also apply here. 
     A further object of the present invention is the use of EGCG for the manufacture of a composition for improving
         the desire to exercise;   the willingness to run and/or perform;   the motivation to move and/or   explorative behavior
 
of animals, preferably of mammals.
       

     A further object of the present invention is the use of EGCG for the manufacture of a composition for improving
         the number of oxidative (“slow-twitch”) muscle fibers;   the number of aerobic skeletal muscle fibers which can help to perform endurance exercise and/or   fiber type shift towards aerobic skeletal muscle fibers
 
in animals, preferably in mammals.
       

     A further object of the present invention is the use of EGCG for the manufacture of a composition for
         accelerating training success;   leading to a faster training success;
 
of animals, preferably of mammals.
       

     The preferences and explanations mentioned above also apply here. 
     When the composition is prepared in form of tablets, capsules, granules or powder for oral administration, there may be used excipients such as lactose, sucrose, sodium chloride, glucose, urea, starch, dextrins and/or maltodextrins, calcium carbonate, calcium phosphate and/or calcium hydrogen phosphate, kaolin, crystalline and/or microcrystalline cellulose and/or silicic acid as carriers; binders such as water, ethanol, propanol, simple syrup, glucose solution, starch and/or hydrolyzed starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropylstarch, shellac, methylcellulose, ethylcellulose, calcium phosphate and/or polyvinyl pyrrolidone; disintegrators such as dry starch, croscarmellose, crospovidone, sodium alginate, agar powder, laminaran powder, sodium hydrogen-carbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch and/or lactose; disintegration-preventing agents such as stearic acid, cacao butter and/or hydrogenated oils; absorbefacients such as quaternary ammonium bases and/or sodium lauryl sulfate; humectants such as glycerol and/or starch; adsorbents such as starch, lactose, kaolin, bentonite and/or colloidal silica; lubricants such as purified talc, stearic acid salts, boric acid powder and/or polyethylene glycol; taste corrigents such as sucrose, orange peel, citric acid and/or succinic acid; and the like. 
     When the composition is prepared in the form of tablets, these may be provided as tablets coated with usual coatings, for example, sugar-coated tablets, gelatin-coated tablets, enteric coated tablets, film-coated tablets, double coated tablets, multilayer-coated tablets and the like. The capsules are prepared by mixing the compounds according to the present invention with the various carriers exemplified above or according to the current state of the art and charging the mixture into hard gelatin capsules, soft capsules and the like. 
     A multi-vitamin and mineral supplement may be added to the compositions according to the present invention, e.g. to maintain a good balanced nutrition or to obtain an adequate amount of an essential nutrient missing in some diets. The multi-vitamin and mineral supplement may also be useful for disease prevention and protection against nutritional losses and deficiencies due to lifestyle patterns and common inadequate dietary patterns sometimes observed in diabetes. 
     The composition according to the present invention can be a food or beverage composition. Beverages can be e.g. sports drinks, energy drinks or other soft drinks, or any other suitable beverage preparation, e.g. joghurt drinks, hot beverages or soups. 
     In a preferred embodiment of the present invention the beverage is an “instant beverage”, i.e. a beverage which is produced—normally by the consumer themself—by stirring a powder into a liquid, usually milk or water. 
     By “sports drink” a beverage is meant which is consumed before, during and/or after physical exercise, mainly to (re-)hydrate as well as to (re-)store electrolytes, sugar and other nutrients. Sports drinks are usually isotonic, meaning they contain the same proportions of nutrients as found in the human body. 
     Energy drinks are beverages which contain (legal) stimulants, vitamins (especially B vitamins) and/or minerals with the intent to give the user a burst of energy. Common ingredients include caffeine, guarana (caffeine from the Guarana plant) and/or taurine, various forms of ginseng, maltodextrin, inositol, carnitine, creatine, glucuronolactone, coenzyme Q10 and/or ginkgo biloba. Some may contain high levels of sugar, or glucose, whereas others are sweetened completely or partially with a sugar alcohol and/or an artificial sweetener like Ca-cyclamate or Aspartame. Many such beverages are flavored and/or colored. 
     A soft drink is a drink that does not contain alcohol. In general, the term is used only for cold beverages. Hot chocolate, tea, and coffee are not considered soft drinks. The term includes carbonated and non-carbonated drinks, e.g. mineral water or so-called “near water drinks”, i.e. water-based beverages which have an additional benefit, e.g. a special flavor and/or further (functional) ingredients. One simple example for a “near water drink” is a mixture of water with very little juice. 
     When the composition is prepared in form of one of the following food articles it is according to the present invention advantageous if the amount of EGCG in the composition is selected from the ranges given in the following table: 
     
       
         
           
               
               
               
             
               
                   
               
               
                   
                   
                 mg EGCG per 
               
               
                   
                   
                 L beverage 
               
               
                 Food Category 
                 typical serving size 
                 or kg food 
               
               
                   
               
             
            
               
                 Beverages (final product) 
                 250 mL, preferred 3 
                 20 to 600, 
               
               
                 (e.g. soft drinks, juices (fruit 
                 servings a day 
                 preferred 
               
               
                 and/or vegetable), teas, 
                   
                 100 to 400 
               
               
                 soups) 
               
               
                 Dairy Products 
                 150 g 
                 170 to 1&#39;000 
               
               
                 (e.g. milk shakes, joghurts, 
               
               
                 joghurt drinks, ice creams) 
               
               
                 Sweets and confectionaries 
                 1 to 5 pieces of 5 g 
                 500 to 10&#39;000, 
               
               
                 (e.g. chocolates, candies, 
                 each per day = 
                 preferred 
               
               
                 mints, jellies, cookies) 
                 5 to 25 g 
                 500 to 1&#39;000 
               
               
                 other food items 
                 25 g 
                 400 to 4&#39;000, 
               
               
                 (e.g. breakfast cereals, 
                   
                 preferred 
               
               
                 muesli bars, snacks, pasta 
                   
                 400 to 1&#39;400 
               
               
                 sauces) 
               
               
                   
               
            
           
         
       
     
     If the composition is prepared in form of a tablet or a capsule it is according to the present invention advantageous if the amount of EGCG is selected from the ranges given in the following table: 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                 Category 
                 typical dosage 
                 mg EGCG per tablet/capsule 
               
               
                   
                   
               
             
            
               
                   
                 Tablets 
                 1-2 × per day 
                 50 to 150 
               
               
                   
                 Capsules 
                 1-2 × per day 
                 50 to 150 
               
               
                   
                   
               
            
           
         
       
     
     The invention is further illustrated by the following examples. 
    
    
     EXAMPLE 
     Example 1 
     Male C57BL/6J mice (5 weeks old) were purchased from Charles River (Sulzfeld, Germany). All mice initially received a high-fat diet (Kliba#2154, Provimi Kliba AG, Kaiseraugst, Switzerland) for 3 months. Thereafter, mice were randomly assigned to 5 groups.
         Group 1: Low-fat diet, restricted to cage normal activity (Low fat sedentary, n=11)   Group 2: High-fat diet, restricted to cage normal activity (High fat sedentary, n=11)   Group 3: High-fat diet containing 0.5% w/w EGCG, restricted to cage normal activity (High fat+EGCG sedentary, n=10)   Group 4: High-fat diet, free access to running wheel (High fat exercise, n=10)   Group 5: High-fat diet containing 0.5% w/w EGCG, free access to running wheel (High fat+EGCG exercise, n=10)       

     The duration of the treatment was 2 months. During this period all mice received food and water ad libitum. Mice in groups 4 and 5 had unlimited access to running wheels (Technical &amp; Scientific Equipment GmbH, Bad Homburg, Germany). Daily voluntary running wheel activity was monitored. At the end of the treatment period, maximal running performance on a rodent treadmill (Technical &amp; Scientific Equipment GmbH, Bad Homburg, Germany) was determined. The Gastrocnemius muscles were excised, frozen in liquid nitrogen and stored at −80° C. for later quantification of the fiber type. In the medial gastrocnemius, skeletal muscle fibers were quantified as Type I, Type IIA, Type D/X, Type IIB by immune-histochemistry utilizing fiber specific antibodies and by histochemistry using a cytochrome oxidase staining. 
     The voluntary running wheel activity of mice receiving the high-fat diet containing EGCG was significantly increased compared to mice receiving the high-fat diet alone: Mice consuming EGCG containing high-fat diets ran on average 3140.3±43.3 m per day while mice on the high fat diet ran only 1579.8±59.0 m per day (P&lt;0.004). 
     Supplementation of the high-fat diet with EGCG increased the percentages of Type I and Type IIa fibers in the medial gastrocnemius muscle of sedentary and exercised mice compared to sedentary mice receiving the high-fat diet alone (Table 1). Type IId/x and Type IIb fibers were less prevalent in the medial gastrocnemius muscle of mice receiving EGCG in the high-fat diet compared to mice receiving the high-fat diet alone (Table 1). The observed fiber type shift in both EGCG groups was more pronounced then the fiber type shift observed in the exercised high-fat group (Table 1). Exercise alone resulted in an increased proportion of Type IIa fibers (Table 1). 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Percentage of fibers in medial gastrocnemius muscle of 
               
               
                 mice receiving the respective treatments for 2 months. 
               
            
           
           
               
               
               
               
               
            
               
                 Group 
                 Type I 
                 Type IIa 
                 Type IId/x 
                 Type IIb 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 Low fat sedentary 
                 7.5 
                 40.8 
                 9.4 
                 42.3 
               
               
                 High fat sedentary 
                 7.8 
                 42.3 
                 10.7 
                 39.2 
               
               
                 High fat + EGCG sedentary 
                 12.5 
                 53.5 
                 5.2 
                 28.8 
               
               
                 High fat exercise 
                 6.3 
                 49.8 
                 5.7 
                 38.1 
               
               
                 High fat + EGCG exercise 
                 12.8 
                 54.2 
                 5.1 
                 27.8 
               
               
                   
               
            
           
         
       
     
     Example 2 
     Pharmaceutical Compositions 
     Soft Gelatine Capsule: 
     The soft gelatine capsule is prepared by conventional formulation procedures using the ingredients specified below: 
     Active ingredient: EGCG 150 mg and vitamin E 50 mg
 
Other ingredients: glycerol, water, gelatine, vegetable oil.
 
     Hard Gelatine Capsule: 
     The hard gelatine capsule is prepared by conventional formulation procedures using the ingredients specified below: 
     Active ingredient: EGCG 300 mg 
     Other Ingredients:
         Fillers: lactose, cellulose and/or cellulose derivatives.   Lubricant: magnesium stearate if necessary (0.5% by weight)       

     Tablet: 
     The tablet is prepared by conventional formulation procedures using the ingredients specified below: 
     Active ingredient: EGCG 300 mg and vitamin E 20 mg
 
Other ingredients: microcrystalline cellulose, silicone dioxide, magnesium stearate, crosscarmellose sodium.
 
     Example 3 
     Food Items 
     Instant Drink: 
     Composition: 
       
     
       
         
           
               
               
             
               
                   
                   
               
               
                   
                 Parts 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 Xylitol 
                 669.4 
               
               
                   
                 Citric acid, anhydrous 
                 220.0 
               
               
                   
                 Carboxymethyl cellulose 
                 30.0 
               
               
                   
                 Tri-sodium-citrate 
                 22.0 
               
               
                   
                 Tri-calcium-phosphate 
                 20.0 
               
               
                   
                 Orange flavour 
                 20.0 
               
               
                   
                 Ascorbic acid 
                 8.0 
               
               
                   
                 Sweetener Twinsweet 
                 4.0 
               
               
                   
                 EGCG as TEAVIGO ™ 
                 6.6 
               
               
                   
                 Powder mix total 
                 1000.0 
               
               
                   
                   
               
            
           
         
       
     
     Preparation: 
     Sieve all ingredients through a 500 μm sieve. Give the powder in an appropriate container and mix on a tumbler blender for at least 20 min. Use 35 g powder for one litre of beverage by adding water. 
     The instant drink contains 50 mg EGCG per serving of 240 ml ready drink. 
     Up to 3 servings per day is recommended. 
     Mints: 
     Composition: 
       
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                 Potency/mint 
                 Amount/mint 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                 EGCG (as TEAVIGO ™ TG) 
                 10 mg 
                 11.1 
                 mg 
               
               
                 Vitamin C as Ascorbic Acid fine granular 
                   
                 10.5 
                 mg 
               
               
                 Sorbitol as Neosorb 60 W 
                   
                 163.6 
                 mg 
               
               
                 Silicon dioxide (Aerosil ™ 200) 
                   
                 1 
                 mg 
               
               
                 Aroma Frescoforte Permaseal 60470-31 
                   
                 10 
                 mg 
               
               
                 (Givaudan) 
               
               
                 Aroma Eiszucker Permaseal 60153-73 
                   
                 6.0 
                 mg 
               
               
                 (Givaudan) 
               
               
                 Sweetener as Twinsweet 
                   
                 1.6 
                 mg 
               
               
                 PEG 6000 
                   
                 20.0 
                 mg 
               
               
                 Magnesium stearate 
                   
                 1.2 
                 mg 
               
               
                 Total Weight 
                   
                 225.0 
                 mg 
               
               
                   
               
            
           
         
       
     
     Preparation: 
     Mix TEAVIGO™ TG, Ascorbic Acid fine granular, Sorbitol, the aromas, sweetener and PEG 6000 into a drum and mix for 10 minutes with a tumbler mixer. Sieve Sorbitol and silicon dioxide through a 1 mm sieve and mix it in a separate drum for 10 minutes. Combine the two powder mixtures and mix again for 10 minutes. Add Magnesium stearate after sieving through a 1 mm sieve and mix for 2 minutes. 
     The powder is compressed to tablets with a Korsch XP1 tablet press, punch size 8 mm round. 
     Up to 5 mints per day are recommended. 
     Soft Drink (Near Water): 
     Typical serving: 250 ml
 
Active ingredient: EGCG: 5-500 mg/per serving
 
     TEAVIGO™, TEAVIGO™ TG: Tradeproducts of DSM Nutritional Products.