MX2008010657A - Compositions and methods for inducing bone growth and inhibiting bone loss - Google Patents

Abstract

Compositions useful for inducing bone growth or inhibiting bone loss in an animal comprising one or more isoflavones or isoflavone metabolites and methods for inducing bone growth or inhibiting bone loss in an animal utilizing such compositions. The compositions and methods are particularly useful for post-menopause, post-andropause, gonadectomized, spayed, or neutered animals.

Description

COMPOSITIONS AND METHODS TO INDUCE BONE GROWTH AND INHIBIT BONE LOSS BACKGROUND OF THE INVENTION FIELD OF THE INVENTION The invention relates generally to compositions and methods for inducing bone growth or inhibiting bone loss and particularly to the use of isoflavones or metabolites thereof to induce bone growth or to inhibit bone loss.

DESCRIPTION OF THE RELATED ART Bone remodeling is a continuous cycle. The cycle begins with bone resorption mediated by the osteoclasts and is followed by the restoration of bone mass by the osteoblasts. The process of bone remodeling is regulated mainly by sex hormones, especially estrogen. However, genetic, nutritional and environmental factors can influence bone loss. It has been shown that estrogen plays an important role in bone remodeling in men as well as in women. Estrogen reduces bone remodeling by suppressing osteoclastogenesis and osteoblastogenesis of bone marrow precursors, it inhibits bone resorption by reducing pro-resorptive cytokines and regulates the lifetime and activities of osteoblasts. Frequently the deregulation of the bone remodeling cycle occurs. A faster rate of bone tissue dissolution and loss is commonly observed than restoration of bone tissue among the elderly population. Pathological bone loss is called osteoporosis. Accelerated bone loss and osteoporosis disproportionately affect women. It is widely accepted that estrogen deficiency, caused by the onset of menopause in women, is a primary contributor to such bone loss. However, bone loss and osteoporosis are seen in men. Although in men there is no physiological equivalent for menopause, many men experience a decrease in sexual hormone circulation and hypogonadism associated with age. These changes are called andropause. Hypogonadism and a decrease in estrogen contribute to bone loss and osteoporosis in men. In this way, both menopause and andropause are risk factors for bone loss. In addition to menopause and andropause, the Surgical removal of sexual organs affects levels of sex hormones such as estrogen. Such removal can affect bone size, mass and density. In animals, procedures such as capping, castration by removing the ovaries, ovariectomy, castration and the like are frequently performed for the control of the population. The practical effect of a gonadectomy on mature female animals is the surgical equivalent of menopause that occurs naturally in older female animals because the procedure effectively decreases circulating levels of sex hormones. After menopause, extragonadal estrogen biosynthesis in female animals depends on the availability of precursor spheroids of the adrenal cortex. Extragonadal estrogen biosynthesis is also important for the normal function of many tissues and systems that include bones in male animals. Circulating testosterone of the testes seems to be the major precursor for extragonadal estrogen biosynthesis. Male animals maintain sufficient concentrations of circulating testosterone throughout life to support extragonadal estrogen biosynthesis. Therefore, male animals usually do not suffer from osteoporosis until they are very old. However, the surgical removal of sexual organs in male animals Mature leads to the complete loss of the production of androgens and estrogens by the testicles. Such elimination also results in the loss of the major precursor for extragonadal estrogen biosynthesis that naturally does not occur in male animals. Bone strength is very dependent on bone density and bone quality. In humansIf peak bone mass is not reached in childhood and adolescence, there is a risk of osteoporosis later in life. Surgical removal of the sex organs in immature and growing male and female animals prevents the animals from reaching peak bone mass by reducing the accumulation of bone mineral density and content. As such, a gonadectomy can be considered as a risk factor for impaired bone growth and development in young growing animals. Hormone replacement therapy and dietary supplement are often used to combat the effects of reduced sex hormone circulation on bone remodeling, particularly on bone loss. With regard to dietary supplementation, dietary phytoestrogens can improve bone mass and bone loss and play a role in osteogenesis. In addition, it is believed that phytoestrogens Dietary supplements have beneficial effects by decreasing or inhibiting bone loss. However, almost all studies related to the beneficial effects of phytoestrogens such as soy isoflavones on bones were conducted either in female animals under the conditions of surgically induced menopause or in postmenopausal women. There is a paucity of data regarding the effects of isoflavones on bone growth in growing animals and in male animals. Phytoestrogens are chemical products made by plants that have a structure similar to mammalian oestrogens. Phytoestrogens are subdivided into three major classifications, that is, cumestans, lignans and isoflavones. Isoflavones have been shown to have positive effects on bone health. Given the risk of (1) impaired bone growth and development and (2) accelerated bone loss and osteoporosis in animals that are in menopause or andropause or that have been gonadectomized, especially when the animals are growing and their skeletal system has not matured yet, there is a need for new compositions and methods that promote healthy bone growth and that reduce or inhibit bone loss in animals without the risk of harmful side effects associated with traditional hormone replacement therapy.

SUMMARY OF THE INVENTION Therefore, an object of the present invention is to provide compositions and methods for inducing bone growth or inhibiting bone loss. Another object of the present invention is to provide compositions and methods for inducing bone growth or inhibiting bone loss in a postmenopausal, post-andropaese, gonadectomized, castrated or layered animal. A further object of the invention is to provide articles of manufacture in the form of kits containing combinations of the isoflavones or their metabolites of the present invention, food compositions, compounds and devices that are useful for inducing bone growth or inhibiting bone loss in an animal . One or more of these other objects are achieved using new compositions and methods to induce bone growth or inhibit bone loss. Generally, the compositions comprise one or more isoflavones or their metabolites in amounts effective to induce bone growth or inhibit bone loss. Isoflavones include at least one of daidzein, 6-0-malonyl-daidzein, 6-0-acetyldaidzein, genistein, 6-0-malonyl-genistein, 6- O-acetyl-genistein, glycitein, 6-0-malonyl-glycitein, 6- O-acetyl-glycitein, biocanin A or formononetin. Also, the isoflavones or their metabolites are soy isoflavones or their metabolites such as equol. The compositions may comprise additional ingredients such as substances that promote or sustain overall healthy bone growth or inhibit bone loss such as DT56a, androstenedione, dehydroepiandrosterone (DHEA), silicone, conjugated linoleic acid (CLA), or orthosilicic acid. Other objects, features and additional advantages of the present invention will be readily apparent to those skilled in the art.

DETAILED DESCRIPTION OF THE INVENTION DEFINITIONS The following abbreviations can be used herein: CLA, conjugated linoleic acid; BCS, body condition score; BMR, basal metabolic index; MER, maintenance energy requirement; DEXA, double energy x-ray absorptiometry; BMC, bone mineral content; BMD, bone mineral density; BW, body weight; SGM, soybean meal.

The term "animal" means a human or other animal, including birds, bovines, canines, equines, felines, pigs, murines, sheep and porcine animals, which could benefit from the induction of bone growth or the inhibition of bone loss. The term "agents that influence the bones" means any compound, composition or drug useful for inducing bone growth or inhibiting bone loss in an animal., different from the isoflavones or their metabolites of the present invention, for example, bisphosphonates, raloxifene, estrogen, calcitonin, risedronate and alendronate. The term "bone growth" means any increase in bone cells or tissues, increase in bone mass, increase in bone minerals, increase in bone density, increase in bone length or increase in bone width, as measured by any adequate means in the art. The term "bone loss" means any decrease in bone cells or tissues, decrease in bone mass, decrease in bone minerals or decrease in bone density in a subject, as measured by any suitable means in the art. The term "conjugated linoleic" or "CLA" is a collective term used to designate a mixture of positional and geometric isomers of linoleic acid of the essential fatty acid (n-6). The term "companion animal" means any domesticated animal, including, without limitation, cats, dogs, rabbits, guinea pigs, ferrets, hamsters, mice, gerbils, horses, cows, goats, sheep, donkeys, pigs and the like. The term "complete and nutritionally balanced pet food" means a pet food that contains all the required nutrients known in appropriate amounts and proportions based on the recommendations of the recognized authorities in the field of pet nutrition and that therefore they have the capacity to serve as a single source of dietary consumption to maintain life or promote production without the addition of supplementary nutritional sources. The nutritionally balanced pet food compositions are well known and widely used in the art. The term "dietary supplement" means a product that is intended to be ingested in addition to the normal diet of an animal. The term "effective amount" means an amount of a compound, material, composition and / or dosage form as described herein that can be effective to achieve a particular biological result. Such results may include, without restriction, induction of healthy bone growth in young, gonadectomized growing animals and inhibition of bone loss in adult animals that are postmenopausal or post-andropausal or that have been gonadectomized. Such effective activity can be achieved, for example, by causing the ingestion of compositions of the present invention. The term "gonadectomized" means an animal whose reproductive organs (testes or ovaries) have been surgically removed. The term "food composition for humans" means any composition intended for ingestion by a human. The term "together" means that an isoflavone or its metabolites, food composition, agents that influence the bones or other compound or composition of the present invention, are administered to an animal (1) together in a food composition or (2) separately at the same or different frequency using the same or different administration routes at approximately the same time or periodically. "Periodically" means that the agent is administered in an acceptable dosage schedule for a specific agent and that the food is provided to an animal routinely as appropriate for the particular animal. "Approximately the same time" generally means that the food and the agent are administered at the same time or within about 72 hours of each other. "Taken together" specifically includes administration schemes wherein the agents influencing the bones are administered for a prescribed period and the compositions comprising one or more isoflavones or their metabolites are administered indefinitely. The term "isoflavones" means 3-phenylchromones, isomeric forms of flavones wherein the benzene group is coupled to the 3-position of the benzopyran ring instead of the 2-position, and their respective metabolites. Whenever the term "isoflavones" is used herein, it is intended that they encompass derivatives and metabolites of isoflavones, with particular examples of isoflavone derivatives as described herein. Isoflavones can be found in various sources, including, without restriction, soy. Non-limiting examples of isoflavones include daidzein, 6-0-malonyl-daidzein, 6-0-acetyldaidzein, genistein, 6-0-malonyl-genistein, 6-0-acetyl-genistein, glycitein, 6-0-malonyl-glycitein, 6-0-acetyl-glycitein, biocanin A, formononetin or some isoflavone metabolites. Isoflavones are known and their use for various benefits to health. For example, it has been discovered that soy reduces the risk of cardiovascular disease; reduces the risk of breast and prostate cancer; relieves hot flashes associated with estrogen deficiency during menopause; retards osteoporosis in postmenopausal women; reduces the total amount of cholesterol, LDL cholesterol and triglycerides in plasma; preserves cognitive functions in postmenopausal women; improves the symptoms of hypertension; and promotes weight loss. The term "L-carnitine" means a trimethylammonium derivative (betaine) of α-amino-hydroxybutyric acid, formed of N 8, 8, 8-trimethyl-lysine and β-butyrobetaine. L-carnitine is an acyl carrier for the mitochondrial membrane that stimulates the oxidation of fatty acid. It is sometimes referred to as Vitamin Bt or Vitamin B7. The term "layered" means an animal that lacks or that has developed imperfectly or its reproductive organs are not functional, whether such a condition occurs congenitally, by natural development processes or through intervention surgery. The term "oral administration" or "oral administration" means that an animal orally ingests a food or other composition.

The term "pet food" or "pet food composition" means a composition that is intended for ingestion by an animal, and preferably by companion animals. The term "simple package" means that the components of a piece of equipment are physically associated in or with one or more containers and that they are considered a unit for manufacture, distribution, sale or use. The containers include, without restriction, bags, boxes, bottles, shrink containers, stapled or otherwise fixed components, or combinations thereof. A simple package can be containers of individual food compositions, physically associated in such a way that they are not considered a unit for manufacturing, distribution, sale or use. The term "castrate" means the removal of the ovaries of a female animal. The term "virtual container" means that the components of a device are associated by addresses on one or several components of virtual or physical equipment that instruct the user on how to obtain the other components, for example, in a bag containing a component and addresses that instruct the user to go to a website, contact a recorded message, observe a visual message, or contact a manager or instructor for instructions on how to use the equipment. All percentages expressed herein are by weight of the composition on a dry matter basis, unless specifically stated otherwise. The term "dry matter base" means that a concentration of the ingredient in a composition is measured after any moisture in the composition is removed. As used throughout this specification, the intervals are used to describe each value witthe range. Any appropriate value witthe range can be selected as the upper or lower value for the interval. The invention is not limited to the particular methodology, protocols and reagents described herein, because they may vary. In addition, the terminology used herein is given for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention. As used herein and in the appended claims, the singular forms "a", "an" and "the" or "the" include the plural reference, unless the context clearly dictates otherwise. Similarly, the words "comprise", "comprise" and "comprising" are to be interpreted inclusively rather than exclusively.

Unless defined otherwise, all technical and scientific terms and some acronyms used herein have the same meanings as those commonly understood by one of ordinary skill in the art in the field of the invention. Although some compositions, methods, articles of manufacture or other means or materials similar or equivalent to those described herein may be used in the practice of the present invention, preferred compositions, methods, articles of manufacture or other means or materials are described at the moment. All patents, patent applications, publications and other references cited or referred to herein are incorporated herein by reference to the extent permitted by law. The discussion of these references is intended only to summarize the assertions made here. No admission is made that any such patents, patent applications, publications or references, or any portion thereof is prior art relevant to the present invention and the right to challenge the accuracy and relevance of such patents, patent applications, publications and other references, is specifically reserved. THE INVENTION In one aspect, the present invention provides compositions suitable for inducing bone growth or inhibiting bone loss in an animal. The compositions comprise one or more isoflavones or their metabolites in an amount effective to induce bone growth or to inhibit bone loss in an animal. The invention is based on the discovery that isoflavones and several of their metabolites are effective in promoting bone growth and bone development, healthy and to inhibit bone loss in animals. The invention is useful for promoting healthy bone growth in young growing animals and reducing bone loss in mature animals. The compositions have been found to be particularly effective in inducing bone growth or inhibiting bone loss in a postmenopausal, post-andropaese, gonadectomized, castrated or coated animal, especially when the skeletal system of the animal is still growing or maturing. The isoflavones or their metabolites may be present in the composition as an ingredient or additive. In one embodiment, the isoflavones are soy isoflavones. In another, the isoflavones are daidzein, 6-0-malonyl-daidzein, 6-0-acetyl-daidzein, genistein, 6-0-malonyl-genistein, 6-0-acetyl-genistein, glycitein, 6-0-malonyl- glycitein, 6-0-acetyl-glycitein, biocanin A or formononetin or metabolites thereof. In a preferred embodiment, the metabolite of isoflavones is dihydrodaidzein or equol. In one embodiment, the compositions further comprise L-carnitine and / or conjugated linoleic acid. In one embodiment, the compositions are pet food compositions. Such compositions include foods designed to supply the necessary dietary requirements for an animal, animal snacks (eg, buns) or dietary supplements. The compositions may be a dry composition (eg, crumbled biscuit), semi-moist composition, wet composition or any mixture thereof. In another embodiment, the composition is a dietary supplement such as a sauce, drinking water, beverage, yogurt, powder, granule, paste, suspension, chewing product, snack, snack, snack, pellet, pill, capsule, tablet or any another form of adequate administration. The dietary supplement can comprise a high concentration of isoflavones or their metabolites so that the supplement can be administered to the animal in small amounts, or alternatively, it can be diluted prior to administration to an animal. The dietary supplement may require mixing with water before administration to the animal. The composition can be refrigerated or frozen.

The isoflavones or their metabolites can be premixed with the other components of the composition to provide the necessary beneficial amounts, can be coated on a pet food composition, or can be added to the composition before being offered to the animal, for example, using a dusted powder or a mixture. In one embodiment, the compositions of the invention comprise isoflavones or their metabolites in an amount effective to induce bone growth in an animal. In another, the compositions comprise isoflavones or their metabolites in an amount effective to inhibit bone loss in an animal. Preferably, in these embodiments, the composition comprises from about 0.1% to about 100% isoflavones or their metabolites, more preferably from about 0.1% to about 50%. For food compositions, the composition preferably comprises from 0.01% to about 50%, preferably from about 0.01% to about 30%. In various embodiments, the composition comprises approximately 1.0%, 1.5%, 2.0%, 2. SJ 9o-, 3.0%,, 3. 4.0%, 4.5%, 5.0%, 5. D ^ 2¾-, 6. 0%, 6.5%, 7. 0%, 7. S So-, 8.0%, 8 • 5%, 9.0%, 9.5%, 10.0%, 10 .5%, ii .0%, 11.J o, 12 .0%, 12. C. o. f 13. 0%, 13.5%, 1 .0%, 14. J¾ / 15. .0%, 15. RS¾-, 16 .0%, 16. C. o- * J f 17. 0%, 17.5%, 18.0%, 18. 19. .0%, 19. Des¾-, .0%, 20. D R S¾- 21. 0%, 21.5¾, 22.0%, 22. j Reo¬, 23. .0%, 23. or f 24. 0%, 24.5%, 25.0%, 25.5%, 26.0%, 26.5%, 27.0%, 27.5%, 28.0%, 28.5%, 29.0%, 29.5%, 30%, 40%, 50% or more of the composition. For example, dietary supplements may be formulated to contain several times or higher concentrations of isoflavones or their metabolites than typical compositions such that the supplements are suitable for administration to an animal in the form of a tablet, capsule, concentrate. liquid or other similar dosage form, or to be diluted before administrations, such as dilution in water, spray or sprinkle on a pet food, and other similar modes of administration. Such supplements may comprise 100% isoflavones or metabolites, but are often formulated with carriers, excipients and the like. The sources of each of the isoflavones or their metabolites can be any suitable source, synthetic or natural. Preferred sources of isoflavones include any plant containing isoflavones, plant material or plant extract, such as, without restriction, legumes, clovers and kudzu root. Preferred sources of legume isoflavones include chickpeas, lentils, soybeans or any other type of beans or chickpeas that contain isoflavones. You can also use soy flour, soybean meal and Similar. Preferred sources of clover isoflavones include red clover and subterranean clover. Alternatively, the isoflavones or their metabolites can be synthesized again according to any suitable means in the art. L-carnitine is a compound of natural origin that plays an important role in energy production in the body of an animal. Higher concentrations of L-carnitine are found in tissues that use fatty acid as their primary energy source, such as the skeleton and cardiac muscle, compared to other tissues. L-carnitine can stimulate osteoblastic differentiation and may play a role in the suppression of bone loss. The source for L-carnitine can be any animal tissue, including isolated mammalian tissue, fish, birds and the like. Similarly, L-carnitine can be obtained from milk isolated from any mammal. Alternatively, L-carnitine can be synthesized again according to any suitable means in the art. CLA plays a role in bone remodeling and growth. In addition, CLA has been shown to benefit BMD in postmenopausal women. The source for CLA can be any animal tissue, including tissue isolated from mammals, birds, fish and the like. Similarly, CLA can be obtained from milk isolated of any mammal. CLA can also be obtained from plants or vegetable oils such as sunflower oil. Alternatively, the CLA can be synthesized again according to any suitable means in the art. CLA can also be derived from synthetic isomers or synthetic analogs of CLA. The compositions may further comprise agents that influence the bones in amounts effective to induce bone growth or inhibit bone loss in an animal, alone or in combination with the isoflavones or their metabolites of the present invention. In various embodiments, the animal is a human or companion animal such as a post-menopausal, post-andropapic, gonadectomized, castrated or stratified animal. In others, the animal is a young or growing animal in which bone development is occurring. The compositions optionally may comprise supplementary substances such as minerals, vitamins, salts, seasonings, colorants and preservatives. Non-limiting examples of supplemental minerals include calcium, phosphorus, potassium, sodium, iron, chloride, boron, copper, zinc, manganese, iodine, selenium and the like. Non-limiting examples of supplemental vitamins include vitamin A, various B vitamins, vitamin C, vitamin D, vitamin E and vitamin K. Additional dietary supplements may also be included, eg, niacin, acid pantothenic, inulin, folic acid, biotin, amino acids and the like. The compositions may optionally comprise one or more supplementary substances that promote or sustain overall healthy bone growth or that inhibit bone loss. Such substances include, without restriction, DT56a, androstenedione, dehydroepiandrosterone (DHEA), silicone, CLA and orthosilicic acid. In various embodiments, the pet food or pet button compositions comprise from about 15% to about 50% crude protein. The crude protein material may comprise vegetable proteins such as soybean meal, soybean protein concentrate, corn gluten meal, wheat gluten, cottonseed and peanut meal, or animal proteins such as casein, albumin and protein. meat. Non-limiting examples of meat protein, useful herein, include pork, lamb, equine, poultry, fish and mixtures thereof. The compositions may also comprise from about 5% to about 40% fat. The compositions may also comprise a carbohydrate source. The compositions may comprise from about 15% to about 60% carbohydrate. Non-limiting examples of such carbohydrates include grains or cereals such as rice, corn, milo, sorghum, alfalfa, barley, soybeans, rapeseed, oats, wheat and mixtures thereof. The compositions optionally may also comprise other materials such as dehydrated whey and other dairy by-products. The compositions may also comprise at least one source of fiber. A range of soluble or insoluble fibers can be used, as will be known to those of ordinary skill in the art. The source of fiber can be beet pulp (beet), gum arabic, talha gum, psyllium, rice bran, locust bean gum, citrus pulp, pectin, fructooligosaccharide additional to the short chain oligofructose, mananooligofructose, soy fiber , arabinogalactan, galactooligosaccharide, arabinoxylan or mixtures thereof. Alternatively, the fiber source can be a fermentable fiber. It has previously been described that fermentable fiber provides a benefit to the immune system of a companion animal. Fermentable fiber or other compositions known to those skilled in the art, which provide a prebiotic composition for improving the growth of probiotic microorganisms within the intestine, may also be incorporated into the composition to aid in the improvement of the benefit provided by the present invention to the immune system animal. Additionally, probiotic microorganisms such as the Enterococcus, Lactobacillus, Bifidobacterium, or Saccharomyces species, for example, can be added to the composition. In a preferred embodiment, the composition is a complete and nutritionally balanced pet food. In this embodiment, the pet food may be a moist food, a dry food, or a food of intermediate moisture content, as would be recognized by those skilled in the art of formulating and manufacturing pet food. "Wet feed" describes pet food that is typically sold in cans or foil pouches and has a moisture content that typically ranges from about 70% to about 90%. "Dry feed" describes pet food that is similar in composition to wet feed, but contains a limited moisture content, typically in the range of from about 5% to about 15%, and is therefore presented, for example. , like small cookies crumbled like buns. The compositions and dietary supplements can be formulated especially for adult animals, or for older or younger animals, for example, a formulation of "puppy food", "kitten food", or "for adults or Senior. "In general, specialized formulations will comprise the appropriate energy and nutritional requirements for animals at different stages of development or age.Some aspects of the invention are preferably used in combination with a complete and balanced feed. which comprise isoflavones or their metabolites according to certain embodiments of the invention, are preferably used with a high quality commercial feed As used herein, "high quality commercial feed" refers to a diet manufactured to produce digestibility of the key nutrients of 80% or more, as established, for example, in the recommendations of the National Research Council mentioned above for dogs, or the guidelines established by the Association of North American Control Officers for Food Control. similar high nutrient standards for others The expert will understand how to determine the appropriate amount of isoflavones or their metabolites that will be added to a given composition. Such factors that may be taken into account include the type of composition (eg, composition of pet food versus dietary supplement), average consumption of types specific to compositions by different animals and the manufacturing conditions under which the composition is prepared. Preferably, the concentrations of isoflavones or their metabolites that will be added to the composition are calculated based on the animal's energy and nutrient requirements. According to certain aspects of the invention, the isoflavones or their metabolites can be added at any time during the manufacture and / or processing of the composition. This includes, without limitation, as part of the formulation of the pet food composition or dietary supplement or as a coating applied to the pet food composition or dietary supplement. The compositions can be made according to any method suitable in the art. In another aspect, the present invention provides methods for inducing bone growth or inhibiting bone loss in an animal. The methods comprise administering to an animal one or more isoflavones or their metabolites in an amount effective to induce bone growth or to inhibit bone loss in the animal. In several embodiments, the isoflavones are daidzein, 6-0-malonyl-daidzein, 6-0-acetyl-daidzein, genistein, 6-0-malonyl-genistein, 6-0-acetyl-genistein, glycitein, 6-0-malonil -glyciteine, 6-0-acetyl-glycitein, biocanin A or formononetin, or its metabolites. In one embodiment, the metabolite of isoflavones is dihydrodaidzein or equol. In one embodiment, the compositions further comprise L-carnitine and / or conjugated linoleic acid. In various embodiments, the composition is a human food composition, pet food composition or a dietary supplement as described herein. In several embodiments, the isoflavones are daidzein, 6-0-malonyl-daidzein, 6-0-acetyl-daidzein, genistein, 6-0-malonyl-genistein, 6-0-acetyl-genistein, glycitein, 6-0-malonil -glycitein, 6-0-acetyl-glycitein, biocanin A or formononetin, or its metabolites. In one embodiment, the metabolite of isoflavones is dihydrodaidzein or equol. In one embodiment, the methods further comprise administering L-carnitine and / or conjugated linoleic acid in combination with the isoflavones or their metabolites. In another, the methods further comprise administering isoflavones or their metabolites in conjunction with agents that influence the bones in amounts effective to induce bone growth or inhibit bone loss in an animal. In various embodiments, the animal is a human or companion animal such as a dog or cat. In certain modes j > the animal is a post-menopausal, post-andropausal or gonadectomized animal. In others, the animal is a young or growing animal. Isoflavones or their metabolites are administered to the animal using a range of administration routes.

Such routes include, without restriction, oral, intranasal, intravenous, intramuscular, intragastric, transpyloric, subcutaneous, rectal and the like. Preferably, the compositions are administered orally. The administration can be on a basis as needed or as desired, for example, once a month, once a week, daily or more than once a day.

Similarly, the administration may be every two days, weeks or months, every three days, weeks or months, every four days, weeks or months, and the like. The administration can be multiple times per day. When used as a supplement to ordinary dietary requirements, the composition may be administered directly to the animal or otherwise contacted with or mixed with the daily food or feed. When used as a daily food or feed, administration will be perfectly known to those with ordinary experience. The administration can also be carried out as part of a diet for the animal. For example, a dietary regimen may comprise causing the regular ingestion by the animal of a composition that it comprises one or more isoflavones or their metabolites, in an amount effective to induce bone growth in the animal, or in an amount effective to inhibit bone loss in the animal. Regular ingestion may be once a day or two, three, four or more times per day, on a daily basis. The goal of regular ingestion is to provide the animal with the preferred daily dose of isoflavones or their metabolites, as exemplified herein. The preferred daily dose ranges for isoflavones and / or their metabolites from about 5 mg / day to about 5000 mg / day per animal. Preferably, the daily dose of isoflavones and / or their metabolites ranges from about 30 mg / day to about 500 mg / day per animal, and more preferably from about 80 mg / day to about 300 mg / day per animal. The daily dose of isoflavones or their metabolites can be measured in terms of grams of isoflavones or their metabolites per kg of BW of the animal. The daily dose of isoflavones or their metabolites may range from about 0.001 g / kg to about 50 g / kg BW of the animal, although higher or lower doses may be provided. Preferably, the daily dose of isoflavones or their metabolites is from about 0.001 g / kg to about 25 g / kg BW of the animal. More preferably, the daily dose of Isoflavones or their metabolites are from about 0.001 g / kg to about 10 g / kg BW of the animal. More preferably, the daily dose of isoflavones or their metabolites is from about 0.001 g / kg to about 5 g / kg BW of the animal. More preferably, the daily dose of isoflavones or their metabolites is from about 0.001 g / kg to about 1 g / kg BW of the animal. More preferably, the daily dose of isoflavones or their metabolites is from about 0.001 g / kg to about 0.15 g / kg BW of the animal. The preferred daily dose ranges for L-carnitine from about 50 mg / day to about 5000 mg / day per animal. Preferably, the daily dose of L-carnitine ranges from about 80 mg / day to about 500 mg / day per animal, and more preferably from about 100 mg / day to about 300 mg / day per animal. The daily dose of L-carnitine can be measured in terms of grams of L-carnitine per kg of BW of the animal. The daily dose of L-carnitine may range from about 0.001 g / kg to about 50 g / kg BW of the animal, although higher or lower doses may be provided. Preferably, the daily dose of L-carnitine is find from about 0.001 g / kg to about 25 g / kg BW of the animal. More preferably, the daily dose of L-carnitine ranges from about 0.001 g / kg to about 10 g / kg BW of the animal. More preferably, the daily dose of L-carnitine is from about 0.001 g / kg to about 5 g / kg BW of the animal. More preferably, the daily dose of L-carnitine is from about 0.001 g / kg to about 1 g / kg BW of the animal. The preferred daily dose ranges for CLA from about 50 mg / day to about 10000 mg / day per animal. Preferably, the daily dose of CLA ranges from about 500 mg / day to about 6000 mg / day per animal, and more preferably from about 1000 mg / day to about 4000 mg / day per animal. The daily dose of CLA can be measured in terms of grams of CLA per kg of BW of the animal. The daily dose of CLA can range from about 0.001 g / kg to about 50 g / kg BW of the animal, although higher or lower doses may be provided. Preferably, the daily dose of CLA is from about 0.001 g / kg to about 25 g / kg BW of the animal. More preferably, the daily dose of CLA is from about 0.001 g / kg to about 10 g / kg BW of the animal. More preferably, the daily dose of CLA is from about 0.001 g / kg to about 5 g / kg BW of the animal. More preferably, the daily dose of CLA is from about 0.001 g / kg to about 1 g / kg BW of the animal. When formulating the compositions of the present invention, an expert can determine the amounts of the isoflavones or their metabolites and other compounds or ingredients, based on the above dosages and on the characteristics of the animal, for example, the species, age, size, weight, health of the animal and similar. According to the methods of the invention, the administration of the isoflavones or their metabolites, which includes administration as part of a diet, can span a period of time ranging from calving to the adult life of the animal. In a further aspect, the present invention provides suitable equipment for administering a composition comprising one or more isoflavones or their metabolites to an animal. The equipment comprises in separate containers in a single container or in separate containers in a virtual container, as appropriate for the component of the equipment, one or more isoflavones or their metabolites and at least one of (1) one or more ingredients suitable for consumption by an animal, (2) one or more agents that influence the bone, suitable for inducing bone growth or inhibiting bone loss, (3) instructions on how to combine isoflavones or their metabolites and other components of the bone. equipment, particularly for producing a food composition useful for administering isoflavones or their metabolites to an animal and (4) instructions for knowing how to use the isoflavones or their metabolites and other components of the present invention, particularly for the benefit of the animal by inducing the bone growth or the inhibition of bone loss in the animal. When the equipment comprises a virtual container, the equipment is limited to instructions in a virtual environment in combination with one or several physical components of the equipment. The kit contains the isoflavones or their metabolites and other components in an amount effective to induce bone growth or inhibit bone loss in the animal. Typically, the isoflavones or their metabolites and the other suitable components of the equipment (eg, food compositions) are mixed just before consumption by an animal. The equipment can contain the components of the equipment in any of several combinations and / or mixtures. In one embodiment, the equipment contains a package comprising one or more of the Isoflavones or their metabolites and a container of food for consumption by an animal. The equipment may contain additional elements such as a device for mixing the isoflavones or their metabolites or the ingredients or a device for containing the mixture, for example, a bowl for food. In another embodiment, isoflavones or their metabolites are mixed with additional nutritional supplements such as vitamins and minerals that promote good health in an animal. The instructions may be directed and / or inform a consumer, doctor, pet owner, veterinarian, food supplier, and the like that the use of the isoflavones or their metabolites may be effected to induce bone growth or to inhibit bone loss in an animal. The instructions can also be directed to the user so that he knows how to apply or mix the isoflavones or their metabolites to an animal's food or water. In another aspect, the present invention provides a means for communicating information regarding or instructions for one or more of (1) using isoflavones or their metabolites to induce bone growth or to inhibit bone loss in an animal, (2) mix the isoflavones or their metabolites with the other components (food compositions) of the present invention, (3) administering the isoflavones or their metabolites to an animal, alone or in combination with the other elements of the present invention and (4) using the kits of the present invention to administer isoflavones or their metabolites to an animal, particularly for induce bone growth or to inhibit bone loss in an animal. The medium comprises a document, digital storage means, optical storage means, audio presentation or visual presentation containing the information or instructions. In certain modalities, the communication means an unfolded website, visual presentation kiosk, brochure, product label, packaging insert, advertising, flyer, public announcement, audio tape, videotape, DVD, CD-ROM, computer readable chip, card readable on computer, computer readable disk, computer memory, or a combination thereof containing such information or instructions. Useful information includes one or more of (1) methods and techniques for combining and administering isoflavones or their metabolites and / or other components and (2) contacting information for animals or their caregivers to use if they have a question regarding the invention. and its use. Useful instructions include amounts for mixing and the administration of quantities and frequency. The means of communication is useful for instructing the benefits of the use of the present invention and communicating the approved methods for administering the invention to an animal. In another aspect, the present invention provides a method for manufacturing a food composition containing isoflavones or their metabolites, comprising mixing one or more ingredients suitable for consumption by an animal and isoflavones or their metabolites or applying isoflavones or their metabolites on the composition food In a further aspect, the present invention provides food compositions made using this method. In a further aspect, the present invention provides the use of isoflavones or their metabolites to prepare a medicament. In another, the invention provides the use of such isoflavones or their metabolites to prepare a medicament for inducing bone growth or for inhibiting bone loss in an animal. Generally, medicaments are prepared by mixing a compound or composition with excipients, buffers, binders, plasticizers, colorants, diluents, compression agents, lubricants, flavors, wetting agents and other ingredients known to those skilled in the art to be useful to produce medicines and formulate medicines that are suitable for administration to an animal.

EXAMPLES The invention may be further illustrated by the following examples, although it should be understood that these examples are included for purposes of illustration only and are not intended to illustrate the scope of the invention, unless specifically indicated otherwise.

Example 1 Effect of dietary soy isoflavones on bone development in normal, gonadectomized dogs Dogs and diets: Thirty Normal, non-obese, newly gonadectomized retrievers, 4-5 months of age, were used in the study. The puppies were randomly divided into three groups, based on the litters to minimize the genetic influence on musculoskeletal growth and health: Group 1 consisted of ten newly gonadectomized retrievers, males and females, which were fed of a perfectly balanced puppy diet that included 29 to 30% protein and 20% fat (the control diet). Group 2 consisted of ten newly gonadectomized retrievers, males and females, who were fed the control diet supplemented with 10% soybean meal (SGM). Group 3 consisted of ten newly gonadectomized Labrador Retrievers, males and females, which were fed with the control diet supplemented with 10% of SGM, 100 ppm of L-carnitine and 1.5% of CLA. SGM contains the following soy isoflavones: 17% genistein, 52% daidzein and 31% glycitein. All the puppies were fed to maintain their ideal body condition score, 5, by adjusting their food intakes. The duration of the study was 70 weeks. All dogs were given a MER determination before the study. Prior to the study, and every week after the study started until the conclusion of the study, the BW was measured for each animal. Before the study, and bi-monthly after the study was started through the conclusion of the study, the following measurements were taken for each animal: DEXA for body fat, lean body mass, bone chemistry, expanded thyroid profile, whole blood count, BMC , bone mineral content of the lumbar vertebra and bone mineral density of the lumbar vertebra. After 70 weeks of growth study, no differences were observed in BW, lean body mass, or body fat among the three groups. It was also determined that all parameters measured in blood chemistry, expanded thyroid profile and whole blood count were within the normal range. The results related to skeletal growth are shown in Tables 1-4. BMC and BMD were evaluated on the lumbar vertebrae 1-7 of each puppy in each experimental group. The results are shown in Tables 1-4. Generally, the Tables show the effects of isoflavones on BMC and BMD. The puppies were fed a control diet (Ration 1) with 20 to 30% protein and 20% fat, an isoflavone diet (Ration 2) (control diet supplemented with 10% of SGM) or a cocktail diet (Ración 3 ) (control diet supplemented with 10% of SGM, 100 ppm of L-carnitine and 1.5% of CLA). In the Tables, Ration 1 = control, Ration 2 = isoflavones, Ration 3 = isoflavones, CLA and L-carnitine. With reference to the Tables, Table 1 shows the effects of soy isoflavones in the diet on the BMC of the lumbar vertebrae 1-4 in male puppies and growing, gonadectomized puppies (4 months of age) during a study of 70 week growth The puppies were fed to maintain their score of Ideal body condition of 5 by adjusting your food consumptions. With reference to the results, BMC was increased in control dogs over the course of the 70-week study, but BMC in puppies consuming isoflavone or cocktail diets containing isoflavone was found to be superior to BMC in puppies consuming the control diet in almost all the occasions tested and at the end of the study. These data clearly indicate that the high quality, perfectly balanced, existing pup diet failed to promote an optimal increase in BMC in growing male and female gonadectomized puppies. Table 2 shows changes in BMD of the lumbar vertebrae 1-4 in growing male and female gonadectomized puppies (4 months of age) during the 70-week growth study. With reference to the results, the BMD of the lumbar vertebrae 1-4 was increased in the control dogs during the course of the 70-week study, but the dogs that consumed isoflavone or cocktail diets containing isoflavone had a higher BMD than the dogs control. These data show that the high quality, perfectly balanced, existing puppy diet failed to promote an optimal increase in BMD in male and female gonadectomized, growing puppies.

Table 3 shows the effects of soy isoflavones in diet on the BMC of the lumbar vertebrae 4-7 in male and female gonadectomized, growing puppies (4 months of age) during the 70-week growth study. With reference to the results, BMC was increased in control dogs over the course of the 70-week study, but at the end of the study BMC in puppies consuming the isoflavone or cocktail diets containing isoflavone was found to be superior to BMC in puppies that consumed the control diet. Notably, contrary to the results observed in the lumbar vertebrae 1-4, where the BMC was generally the same between dogs fed the cocktail and diets with isoflavone, the results observed in the lumbar vertebrae 4-7 indicate that in the Overall, the dogs fed the cocktail diet had a BMC higher than that of the dogs fed the isoflavone diet. Table 4 shows changes in BMD of the lumbar vertebrae 4-7 in male and female gonadectomized, growing puppies (4 months of age) during the 70-week growth study. With reference to the results, the BMD of the lumbar vertebrae 4-7 was increased in the control dogs in the course of the 70-week study, but the dogs that consumed the isoflavone diets or cocktails that containing isoflavone had a higher BMD than the control dogs at the end of the study when the skeletal system of the dogs reached maturity. These data show that high-quality, perfectly balanced, existing puppy diets could not compensate for the loss of estrogen-promoting effects that were lost. after the gonadectomy. As the results show, the control puppies never achieved BMC and BMD peak optima when their skeletal system reached maturity. Conversely, isoflavones and isoflavone-containing cocktail are effective in promoting better bone growth (BMC and higher BMD) in growing gonadectomized puppies. Table 1 Weeks Ration 1 SE 1 Ration 2 SE 2 Ration 3 SE 3 BMC 13.2741 1.1223 12.6152 1.1223 12.8699 1.0874 L1-L4 16 BMC 19.0018 1.0715 18.9969 1.0717 20.2617 1.0348 BMC 22.1142 1.0637 22.6728 1.0637 23.7819 1.0378 L1-L4 34 BMC 24.2471 1.0434 25.2927 1.0435 25.7862 1.0244 L1-L4 42 BMC 25.7025 1.054 27.0549 1.0545 26.8387 1.0396 L1-L4 50 BMC 26.7823 1.0708 28.1578 1.0717 27.5036 1.0601 L1-L4 58 BMC 27.7883 1.0656 28.7997 1.0658 28.3452 1.0574 L1-L4 66 BMC 28.3579 1.1022 29.0097 1.1022 29.0085 1.0956 Table 2 Weeks Ration 1 SE 1 Ration 2 SE 2 Ration 3 SE 3 BMD 0.7067 0.02817 0.7049 0.02817 0.7341 0.02722 L1-L4 16 BMD 0.826 0.02715 0.836 0.02715 0.8561 0.02616 L1-L4 26 BMD 0.8847 0.02699 0.9072 0.02699 0.9107 0.02624 L1-L4 34 BMD 0.92 0.02659 0.9543 0.02659 0.9399 0.02599 BMD 0.9404 0.0268 0.9831 0.0268 0.9555 0.02631 L1-L4 50 BMD 0.9542 0.02714 0.9992 0.02714 0.9695 0.02675 L1-L4 58 BMD 0.9699 0.02703 1.0081 0.02703 0.9938 0.02673 L1-L4 66 BMD 0.9811 0.02776 1.01 16 0.02776 0.0135 0.0275 L1-L4 70 Table 3 Weeks Ration 1 SE 1 Ration 2 SE 2 Ration 3 SE 3 BMC 14.4363 1.1379 13.5979 1.1379 14.426 1.1001 L4-L7 16 BMC 20.7348 1.0949 20.6968 1.0949 22.369 1.0554 L4-L7 26 BMC 24.4373 1.0883 24.7872 1.0883 26.4567 1.0585 L4-L7 34 BMC 27.1118 1.0711 27.693 1.071 1 29.0178 1.0476 L4-L7 42 BMC 28.9061 1.08 29.6245 1.08 30.5032 1.061 L4-L7 50 BMC 29.9677 1.0944 30.7917 1.0944 31.3636 1.0793 L4-L7 58 BMC 30.4443 1.0899 31.405 1.0899 32.0497 1.0782 L4-L7 66 BMC 30.5093 1.1208 31.5695 1.1208 32.4683 1.1 11 L4-L7 70 Table 4 Weeks Ration 1 SE 1 Ration 2 SE 2 Ration 3 SE 3 BMD 0.6846 0.02579 0.6652 0.02579 0.7117 0.02496 L4-L7 16 BMD 0.799 0.02473 0.8009 0.02473 0.827 0.02385 L4-L7 26 BMD 0.856 0.02456 0.8643 0.02456 0.8821 0.02392 L4-L7 34 BMD 0.8904 0.02414 0.9003 0.02414 0.9145 0.02365 L4-L7 42 BMD 0.9099 0.02436 0.9208 0.02436 0.9335 0.02397 L4-L7 50 BMD 0.9221 0.02471 0.9376 0.02471 0.9488 0.02441 L4-L7 58 BMD 0.9347 0.0246 0.9626 0.0246 0.9697 0.02437 L4-L7 66 BMD 0.9434 0.02537 0.9818 0.02537 0.9852 0.02518 L4-L7 70 In the specification, typical preferred embodiments of the invention have been described and, although specific terms are employed, they are used in a generic and descriptive sense only and not for purposes of limiting the scope of the invention set out in the claims. Obviously, many modifications and variations of the invention are possible in light of the above teachings. Therefore, it should be understood that within the scope of the appended claims, the invention may be practiced otherwise than specifically described.

Claims (24)

1. Composition comprising one or more isoflavones or their metabolites in an amount effective to induce bone growth or to inhibit bone loss in an animal.

2. Composition according to claim 1, wherein the composition is a food composition for human, pet food composition, or a dietary supplement.

3. Food composition according to claim 2, wherein the food composition comprises from about 0.1% to about 50% of isoflavones or their metabolites.

4. Composition according to claim 1, wherein the isoflavones include at least one of daidzein, 6-O-malonyl-daidzein, 6-O-acetyl-daidzein, genistein, 6-O-malonyl-genistein, 6-O-acetyl-genistein , glycitein, 6-O-malonyl-glycitein, 6-O-acetyl-glycitein, biocanin A or formononetin.

Composition according to the claim where the isoflavones or their metabolites are soy isoflavones or their metabolites.

6. Composition according to claim 5, wherein the metabolite of soy isoflavone is equol.

7. Composition according to claim 1, wherein the animal is a dog or a cat.

8. Composition according to the claim wherein the animal is a postmenopausal, andropramatic, gonadectomized, castrated or layered animal.

9. Composition according to claim 1, further comprising at least one of DT56a, androstenedione, dehydroepiandrosterone (DHEA), silicone, L-carnitine, conjugated linoleic acid or orthosilicic acid.

10. Method for inducing bone growth or for inhibiting bone loss in an animal, comprising administering to the animal a composition containing one or more isoflavones or their metabolites in an amount effective to induce bone growth or to inhibit bone loss in the animal .

11. The method according to claim 10, wherein the composition is a food composition for humans, a pet food composition or a dietary supplement.

12. The method according to claim 11, wherein the food composition comprises from about 0.1% to about 50% of isoflavones or their metabolites.

13. The method according to claim 10, wherein the isoflavones include at least one of daidzein, 6-0-malonyl-daidzein, 6-0-acetyl-daidzein, genistein, 6-0-malonyl-genistein, 6-0-acetyl-genistein , glycitein, 6-0-malonyl-glycitein, 6-0-acetyl-glycitein, biocanin A or formononetin.

14. The method according to claim 10, wherein the isoflavones are soy isoflavones or their metabolites.

15. The method according to claim 14, wherein the metabolite of soy isoflavone is equol.

16. Method according to claim 10, wherein the animal is a post-menopausal, post-andropausal animal, gonadectomized, castrated or layered.

17. Method according to claim 10, wherein the composition is administered to the animal daily.

18. The method according to claim 10, wherein the composition further comprises at least one of DT56a, androstenedione, dehydroepiandrosterone (DHEA), silicone, L-carnitine, conjugated linoleic acid or orthosilicic acid.

19. The method according to claim 10, wherein the isoflavones or their metabolites are administered in conjunction with one or more agents that influence the bones.

20. Method according to claim 10, wherein the animal is a dog or a cat.

21. Suitable equipment for administering a composition comprising one or more isoflavones or their metabolites to an animal, comprising in separate containers in a single container or in separate containers in a virtual container, as appropriate for the component of the equipment, one or more isoflavones or its metabolites and less one of one or several ingredients suitable for consumption by an animal, one or several agents that influence the bones, suitable for inducing bone growth or for inhibiting bone loss, instructions on how to combine isoflavones or their metabolites and other components of the equipment, and instructions for knowing how to use the isoflavones or their metabolites and other components of the present invention.

22. Means for communicating information regarding or instructions for one or more of using the isoflavones or their metabolites to induce bone growth or to inhibit bone loss in an animal, mix the isoflavones or their metabolites with the other components of the present invention, administer isoflavones or their metabolites to an animal, alone or in combination with the other elements of the present invention and using the kits of the present invention to administer isoflavones or their metabolites to an animal, comprising a document, digital storage media, means optical storage, audio presentation or visual presentation containing the information or instructions.

Medium according to the claim selected from the group consisting of an unfolded website, visual presentation kiosk, brochure, product label, packaging insert, advertising, flyer, public announcement, audio tape, videotape, DVD, CD-ROM, computer readable chip, card readable on computer, computer readable disk, computer memory, or combination thereof.

24. Method for manufacturing a food composition containing isoflavones or their metabolites, comprising mixing one or more ingredients suitable for consumption by an animal and isoflavones or their metabolites or applying isoflavones or their metabolites on the food composition. 25 Use of isoflavones or their metabolites to prepare a medicament for inducing bone growth or to inhibit bone loss in an animal.